PMID- 19902267 TI - Combination therapy with steroids and mizoribine in juvenile SLE: a randomized controlled trial. AB - The initial treatment of childhood-onset systemic lupus erythematosus (SLE) is not standardized. Although corticosteroids are the first-line therapy for SLE, long-term, high-dose steroid therapy is associated with various side effects in children. The Japanese Study Group for Renal Disease in Children (JSRDC) has carried out a multi-center, randomized, controlled trial to evaluate the efficacy and safety of corticosteroid and mizoribine (MZB) therapy as an initial treatment for newly diagnosed juvenile SLE. Twenty-eight patients were treated with a combination steroid and MZB (4-5 mg/kg/day) (group S+M) drug therapeutic regimen, while 29 patients were treated with steroid only (group S); both groups were followed up for 1 year. The time to the first flare from treatment initiation was not significantly different between the two groups (Kaplan-Meier method, p = 0.09). During the period when the steroid was given daily (day 0-183), the time to the first flare from treatment initiation was significantly longer in the patients of group S+M than in those of group S (log-rank test, p = 0.02). At the end of the study period, there were no differences in the severity of proteinuria and renal function impairment between the two groups. No patients dropped out of the trial due to adverse events. In conclusion, our combined steroid and MZB drug therapeutic regimen was not shown to be significantly better than the steroid only therapy as initial treatment for juvenile SLE. Whether MZB administered in a higher dose would be therapeutically advantageous can only be answered by further studies. PMID- 19902268 TI - Resolution of chyloperitoneum in a preterm with octreotide, diet and cessation of dialysis. AB - The diagnosis of chyloperitoneum (CP) is based on the presence of high levels of triglycerides (TGs) in the dialysate. It is a rare complication of peritoneal dialysis (PD) and even rarer in neonates. We report here the case of CP in a 1700 g male baby delivered at the 30th gestational week due to posterior urethral valve and associated oligohydramnios. On postnatal day 2, the serum creatinine (Scr) was 1.6 mg/dL, and he was anuric. PD was instituted via a Tenckhoff catheter. At the end of the second week, after the initiation of enteral feeding, the ultrafiltrate became cloudy, with a leukocyte count of 900/mm(3). A treatment regimen consisting of intraperitoneal vancomycin and ceftazidime was then started. Five days later, the fluid became milky, with a TG level of 251 mg/dL. The patient was then placed on a diet based on medium-chain triglycerides and octreotide (1 microg/kg/h; increasing up to 2 microg/kg/h over 15 days). Although the TG and leukocyte levels decreased, the milky appearance persisted. PD was stopped for 2 days when the Scr decreased to 1.7 mg/dL. When it was resumed, the fluid was totally clear, with a TG level of 7 mg/dL. The infant was discharged with a nightly intermittent peritoneal dialysis program and has had no recurrence. In summary, we report a preterm infant who developed CP during PD and recovered following treatment that included diet modification, octreotide, and temporary discontinuation of the PD. PMID- 19902269 TI - Temporal and spatial expression of a growth-regulated network of imprinted genes in growth plate. AB - In mammals, the somatic growth rate is rapid during fetal and early postnatal life and then gradually declines and eventually stops. In search of the fundamental biological mechanism causing coordinated growth deceleration in multiple tissues, a network of imprinted genes was recently identified based on a coordinated decline in expression in several organs during postnatal growth. To explore a possible role in longitudinal bone growth, we characterized expression of the network during postnatal growth in microdissected metaphyseal bone and growth plate zones of 1-, 3-, and 9-week-old rats using real-time PCR. The expression pattern of the network is modified in growth plate. Similar to the coordinated decline previously observed in kidney, lung, liver, and heart, expression of all genes, except Gtl2, decreased with age in metaphyseal bone. On the contrary, Mest, Dlk1, H19, and Gtl2 decreased, and Cdkn1c, Grb10, and Slc38a4 increased with age in growth plate. During differentiation from resting to hypertrophic zone, Mest, Dlk1, Grb10, and Gtl2 expression decreased, whereas Slc38a4 expression increased. In particular, developmental changes in the expression of growth-promoting genes, Mest, Dlk1, Gtl2, and growth-inhibitory genes, Cdkn1c and Grb10, may contribute to the decline in longitudinal bone growth that occurs with age. PMID- 19902270 TI - Effects of GH in human muscle and fat. AB - Skeletal muscle is the major constituent of lean body mass and a major determinant of energy expenditure both at rest and during physical activity. Growth hormone, in turn, influences muscle mass as well as energy expenditure. Growth hormone substitution in adults increases muscle mass by 5-10%, but part of the effect is attributed to rehydration rather than protein accretion. In addition, GH regulates substrate metabolism in muscle and in particular antagonizes insulin-stimulated glucose disposal. This effect is linked to increased free fatty acid (FFA) flux but the molecular mechanisms remain unclear. During fasting, GH-induced insulin resistance may be favorable by reducing the demand of gluconeogenesis from protein. But in the postprandial phase, GH exposure may compromise glucose tolerance via the same mechanisms. Understanding the mechanisms whereby GH antagonizes insulin-stimulated glucose disposal in muscle is an important future research field with implications for a variety of clinical conditions ranging from malnutrition to obesity and type 2 diabetes. PMID- 19902271 TI - Quality of life or health status in children with chronic kidney disease. PMID- 19902272 TI - Longitudinal growth in chronic hypokalemic disorders. AB - Growth retardation remains a major complication in children with primary tubular disorders, despite adequate supplemental treatment with electrolytes, water and bicarbonate. Chronic hypokalemia, characteristic of some tubulopathies, impairs growth by mechanisms that are not well known. Association with growth hormone deficiency has been reported in patients with Bartter's or Gitelman's syndrome. Tissue-specific alterations of growth hormone and insulin-like growth factor I axis have been described in experimental models of potassium depletion. Hypokalemic rats gain less body length and weight than pair-fed normokalemic animals and, by contrast, develop renal hypertrophy. These rats have low circulating concentrations of insulin-like growth factor I, depressed messenger ribonucleic acid (mRNA) levels of this peptide in the tibial growth plate, and they are resistant to the longitudinal growth-promoting effects of exogenous growth hormone. The reason for this resistance remains to be defined. No alterations in the intracellular signaling for growth hormone have been found in the liver of hypokalemic rats. However, treatment with high doses of growth hormone is unable to normalize hypertrophy of the epiphyseal cartilage chondrocytes, which are severely disturbed in potassium depletion and likely play an important role in the pathogenia of growth impairment in this condition. PMID- 19902273 TI - Physical activity interests and preferences in palliative cancer patients. AB - GOALS OF THE WORK: The purpose of the present study was to examine the physical activity interests and preferences of palliative cancer patients. PATIENTS AND METHODS: Fifty advanced cancer patients aged 18 years or older with clinician estimated life expectancy of less than 12 months and Palliative Performance Scale greater than 30% were recruited from an outpatient palliative care clinic and a palliative home care program. An interviewer-administered survey was used to assess physical activity behavior, interests, and preferences. MAIN RESULTS: The majority of palliative cancer patients in this sample indicated that they would be interested and felt able to participate in a physical activity program. There were strong preferences for home-based physical activity programs that could be completed alone. The majority of participants in this sample reported walking as the type of physical activity in which they were most interested. CONCLUSIONS: These findings suggest that this sample of palliative cancer patients is interested and feel able to participate in a physical activity program. Examining these unique physical activity preferences may aid in the development of an appropriate physical activity program for palliative cancer patients that can be tested in clinical research. PMID- 19902274 TI - A survey on physician knowledge and attitudes towards clinical use of morphine for cancer pain treatment in China. AB - BACKGROUND: The WHO's three-step guideline for cancer pain management has been introduced in China; however, there remain large differences in the standards of cancer pain management between China and other developed countries. This survey was carried out to determine the degree of physician knowledge on morphine use and the factors that impede morphine use in clinical practice in China. METHODS: A self-reported questionnaire was designed and administered to randomly selected physicians in four tertiary hospitals in the cities of Changchun and Changsha in China. Statistical analyses were conducted using SPSS statistical software. RESULTS: Two hundred and one clinical physicians participated in the survey. Physicians who reported having received training in cancer pain management and drug use demonstrated a significantly higher mean score of basic knowledge compared to physicians who reported not having received training (9.31 +/- 2.88:8.23 +/- 2.70, u = 2.74, p < 0.001). The top three cited impediments to widespread clinical use of morphine for cancer pain were: (1) lack of professional knowledge and training; (2) fear of opioid addiction; and (3) physicians' personal preferences to select other drugs. CONCLUSIONS: Medical staffs lack the basic knowledge and harbor misconceptions about the clinical use of morphine for cancer pain treatment. Creating training opportunities for medical staffs is necessary to increase their awareness and knowledge of effective cancer pain management. PMID- 19902275 TI - Comparative analysis between shape memory alloy-based correction and traditional correction technique in pedicle screws constructs for treating severe scoliosis. AB - The three-dimensional correction of severe rigid scoliosis has been improved by segmental pedicle screw instrumentation. However, there can be significant difficulty related to the use of a rigid rod, especially in the apex region of severe scoliosis. This study is a retrospective matched cohort study to evaluate the advantages of Nitinol shape memory alloy (SMA) rod-based correction by comparing the clinical and radiographic results obtained from using a temporary SMA rod and those from a standard rod in the correction of severe scoliosis. From May 2004 to September 2006, patients with matched curve type, ages at surgery, operative methods and fusion levels in our institute and instrumented with either SMA rods (n = 14) or traditional correction techniques (n = 16) were reviewed. In SMA group, the SMA rods served as a temporary intraoperative tool for deformity correction and were replaced by standard rods. The blood loss at surgery averaged 778 +/- 285 ml in the traditional group and 585 +/- 188 ml in the SMA group (P < 0.05). Operative time averaged 284 +/- 53 min in the SMA group and 324 +/- 41 min in the traditional group (P < 0.05). In the SMA group, the preoperative major curve was 92.6 degrees +/- 13.7 degrees with a flexibility of 25.5 +/- 7.3% was corrected to 29.4 degrees +/- 5.7 degrees demonstrating a 68.4% immediate postoperative correction. In the traditional group, the preoperative major curve was 88.6 degrees +/- 14.6 degrees with a flexibility of 29.3 +/- 6.6% was corrected to 37.2 degrees +/- 7.3 degrees demonstrating a 57.8% immediate postoperative correction. There was a statistic difference between the SMA group and traditional group in correction rate of the major thoracic curve. In the SMA group, one case suffered from deep infection 2 months postoperatively. In the traditional group, 6 of 16 cases suffered pedicle screw pull out or loosening during placement of the standard rod at the apex vertebrae on the concave side. In three cases, the mono-axial pedicle screws near the apex were abandoned and in five cases replaced with poly-axial pedicle screws. This study shows that the temporary use of SMA rod may reduce the operative time, blood loss, while improve the correction of the coronal plane when compared with standard techniques. PMID- 19902276 TI - The impact of intra-operative sternum vertical displacement on the sagittal curves of the spine. AB - Patient positioning is an important step in spinal surgeries. Many surgical frames allow for lumbar lordosis modulation due to lower limb displacement, however, they do not include a feature which can modulate thoracic kyphosis. A sternum vertical displacer (SVD) prototype has been developed which can increase a subject's thoracic kyphosis relative to the neutral prone position on a surgical frame. The kyphosis increase is obtained by lifting the subject's torso off the thoracic cushions with a dedicated sternum cushion that can be displaced vertically. The objective of this study was to evaluate the impact of SVD utilization on the sagittal curves of the spine. Experimental testing was performed on six healthy volunteers. Lateral radiographs were taken in the neutral and sternum raised positions and then analyzed in order to compare the values of sagittal curves. The displacement of volunteers and surgical frame components between positions was recorded using an optoelectronic device. Finally, interface pressures between the volunteers and surgical frame cushions were recorded using a force sensing array. Average results show that passing from the neutral to sternum raised positions caused an increase of 53% in thoracic kyphosis and 24% in lumbar lordosis; both statistically significant. Sensors showed that the sternum was raised a total of 8 cm and that interface pressures were considerably higher in the raised position. The SVD provides a novel way of increasing a patient's thoracic kyphosis intra-operatively which can be used to improve access to posterior vertebral elements and improve sagittal balance. It is recommended that its use should be limited in time due to the increase in interface pressures observed. PMID- 19902277 TI - Plasma disc decompression for contained cervical disc herniation: a randomized, controlled trial. AB - Prospective case series studies have shown that plasma disc decompression (PDD) using the COBLATION SpineWand device (ArthroCare Corporation, Austin, TX) is effective for decompressing the disc nucleus in symptomatic contained cervical disc herniations. This prospective, randomized controlled clinical trial was conducted to evaluate the clinical outcomes of percutaneous PDD as compared to conservative care (CC) through 1 year. Patients (n = 115) had neck/arm pain >50 on the visual analog scale (VAS) pain scale and had failed at least 30 days of failed CC. Patients were randomly assigned to receive either PDD (n = 62) or CC (n = 58). Clinical outcome was determined by VAS pain score, neck disability index (NDI) score, and SF-36 health survey, collected at 6 weeks, 3 months, 6 months, and 1 year. The PDD group had significantly lower VAS pain scores at all follow-up time points (PDD vs. CC: 6 weeks, -46.87 +/- 2.71 vs. -15.26 +/- 1.97; 3 months, -53.16 +/- 2.74 vs. -30.45 +/- 2.59; 6 months, -56.22 +/- 2.63 vs. 40.26 +/- 2.56; 1 year, -65.73 +/- 2.24 vs. -36.45 +/- 2.86; GEE, P < 0.0001). PDD patients also had significant NDI score improvement over baseline when compared to CC patients at the 6 weeks (PDD vs. CC: -9.15 +/- 1.06 vs. -4.61 +/- 0.53, P < 0.0001) and 1 year (PDD vs. CC: -16.70 +/- 0.29 vs. -12.40 +/- 1.26, P = 0.005) follow-ups. PDD patients showed statistically significant improvement over baseline in SF-36 physical component summary scores when compared to CC patients at 6 weeks and 1 year (PDD vs. CC: 8.86 + 8.04 vs. 4.24 +/- 3.79, P = 0.0004; 17.64 +/- 10.37 vs. 10.50 +/- 10.6, P = 0.0003, respectively). In patients who had neck/arm pain due to a contained cervical disc herniation, PDD was associated with significantly better clinical outcomes than a CC regimen. At 1 year, CC patients appeared to suffer a "relapse, showing signs of decline in most measurements, whereas PDD patients showed continued stable improvement. PMID- 19902278 TI - Intraseptal vs. periodontal ligament anaesthesia for maxillary tooth extraction: quality of local anaesthesia and haemodynamic response. AB - There is no data concerning the use of the intraseptal anaesthesia (ISA) for single tooth extraction. The aims of this study were to compare the clinical efficacy and haemodynamic responses of the ISA with the periodontal ligament anaesthesia (PLA) for single tooth extraction. Thirty-five randomly selected healthy patients (ASA I) undergoing maxillary lateral incisors extraction entered the study. Onset of anaesthesia, the width of the anaesthetic field and duration of anaesthesia were recorded by pinprick testing. Intensity of anaesthesia was evaluated on a visual analogue scale. Haemodynamic parameters were recorded simultaneously at different time points after anaesthesia injection. The two techniques of local anaesthesia did not show statistically significant differences regarding the success rate and onset of anaesthesia, while the duration of the ISA on the buccal site was significantly longer in comparison with the PLA. The intensity of the achieved anaesthesia, estimated by the experienced pain during procedure, pointed out that pain was recorded in 24% of cases in the ISA group, and in 19% in the PLA group without significant differences. Postoperative pain was found to be smaller in the ISA group (70.9% of treated sites) than in the PLA group (81.3% of treated sites); however, this difference was not significant. Although the heart rate increased in both groups, there were no significant differences in the patients' haemodynamic response between the ISA and the PLA. The results of the present study indicate that both techniques are useful and suitable for the routine tooth extraction. PMID- 19902279 TI - Rapid identification and antimicrobial susceptibility profiling of Gram-positive cocci in blood cultures with the Vitek 2 system. AB - Rapid identification and antimicrobial susceptibility profiling of the bacteria in blood cultures can result in clinical and financial benefits. Addition of saponin to the fluid from blood culture bottles promotes the recovery of the bacteria and thus may shorten the turnaround time of the microbiological analyses. In this study we compared the identification and susceptibility profiles of saponin-treated and untreated (standard method) blood cultures monomicrobial for Gram-positive cocci using Vitek 2. We concordantly identified 49 (89%) of 55 monobacterial cultures using the results with the standard method as reference. Complete categorical agreement between the susceptibility profiles with the new and the standard method was found for 26 (53%) of 49 isolates, while discrepancies were seen for 23 (47%) cultures. E-tests indicated that the new method resulted in a correct susceptibility profile for 8 (35%) of these 23 blood cultures. Therefore, 34 (69%) of 49 cultures showed a concordant/correct susceptibility profile for all antimicrobials with an overall error rate of 2.3%. Thus, addition of saponin to the fluid from blood culture bottles of the Bactec 9240 leads to the rapid (results available >or=12 hours earlier) and reliable identification and susceptibility profiling of Gram-positive cocci in blood cultures with Vitek 2. PMID- 19902280 TI - The use of Macronet resins to recover gamma-decalactone produced by Rhodotorula aurantiaca from the culture broth. AB - During the biotransformation of castor oil into gamma-decalactone, R. aurantiaca produced both the lactone form and its precursor (4-hydroxydecanoic acid). After six days of culture, a maximum yield of gamma-decalactone of 6.5 g/l was obtained. The parameters of gamma-decalactone adsorption on three Macronet resins (MN-202, MN-102 and MN-100) were investigated in water. Adsorption isotherms of gamma-decalactone for the three Macronet resins were linear. The trapping of gamma-decalactone produced by R. aurantiaca on these resins was then carried out. gamma-Decalactone was effectively retained by all the studied Macronet resins. The resin MN-202 trapped gamma-decalactone more efficiently than MN-102 and MN 100. The percentages of gamma-decalactone adsorbed on the resins MN-202, MN-102 and MN-100 were, respectively, 85, 75 and 81%, whereas around 70% of the adsorbed gamma-decalactone was then desorbed. We propose an industrial process that uses Macronet resins to extract gamma-decalactone from culture broth of R. aurantiaca. PMID- 19902281 TI - Cellulases from Penicillium funiculosum: production, properties and application to cellulose hydrolysis. AB - The objective of this work is to investigate the utilization of two abundant agricultural residues in Brazil for the production and application of cellulolytic enzymes. Different materials obtained after pretreatment of sugarcane bagasse, as well as pure synthetic substrates, were considered for cellulase production by Penicillium funiculosum. The best results for FPase (354 U L(-1)) and beta-glucosidase (1,835 U L(-1)) production were observed when sugarcane bagasse partially delignified cellulignin (PDC) was used. The crude extract obtained from PDC fermentation was then partially characterized. Optimal temperatures for cellulase action ranged from 52 to 58 degrees C and pH values of around 4.9 contributed to maximum enzyme activity. At 37 degrees C, the cellulases were highly stable, losing less than 15% of their initial activity after 23 h of incubation. There was no detection of proteases in the P. funiculosum extract, but other hydrolases, such as endoxylanases, were identified (147 U L(-1)). Finally, when compared to commercial preparations, the cellulolytic complex from P. funiculosum showed more well-balanced amounts of beta-glucosidase, endo- and exoglucanase, resulting in the desired performance in the presence of a lignocellulosic material. Cellulases from this filamentous fungus had a higher glucose production rate (470 mg L(-1) h(-1)) when incubated with corn cob than with Celluclast, GC 220 and Spezyme (312, 454 and 400 mg L(-1) h(-1), respectively). PMID- 19902282 TI - Generation and characterisation of stable ethanol-tolerant mutants of Saccharomyces cerevisiae. AB - Saccharomyces spp. are widely used for ethanologenic fermentations, however yeast metabolic rate and viability decrease as ethanol accumulates during fermentation, compromising ethanol yield. Improving ethanol tolerance in yeast should, therefore, reduce the impact of ethanol toxicity on fermentation performance. The purpose of the current work was to generate and characterise ethanol-tolerant yeast mutants by subjecting mutagenised and non-mutagenised populations of Saccharomyces cerevisiae W303-1A to adaptive evolution using ethanol stress as a selection pressure. Mutants CM1 (chemically mutagenised) and SM1 (spontaneous) had increased acclimation and growth rates when cultivated in sub-lethal ethanol concentrations, and their survivability in lethal ethanol concentrations was considerably improved compared with the parent strain. The mutants utilised glucose at a higher rate than the parent in the presence of ethanol and an initial glucose concentration of 20 g l(-1). At a glucose concentration of 100 g l(-1), SM1 had the highest glucose utilisation rate in the presence or absence of ethanol. The mutants produced substantially more glycerol than the parent and, although acetate was only detectable in ethanol-stressed cultures, both mutants produced more acetate than the parent. It is suggested that the increased ethanol tolerance of the mutants is due to their elevated glycerol production rates and the potential of this to increase the ratio of oxidised and reduced forms of nicotinamide adenine dinucleotide (NAD(+)/NADH) in an ethanol-compromised cell, stimulating glycolytic activity. PMID- 19902283 TI - Complex problem solving: a case for complex cognition? AB - Complex problem solving (CPS) emerged in the last 30 years in Europe as a new part of the psychology of thinking and problem solving. This paper introduces into the field and provides a personal view. Also, related concepts like macrocognition or operative intelligence will be explained in this context. Two examples for the assessment of CPS, Tailorshop and MicroDYN, are presented to illustrate the concept by means of their measurement devices. Also, the relation of complex cognition and emotion in the CPS context is discussed. The question if CPS requires complex cognition is answered with a tentative "yes." PMID- 19902284 TI - Complex problem solving: another test case? AB - Attacks on classic complex problem solving focus on both their ecological validity and the difficulty to analyze such a complex interplay of system variables. But we argue that the domain of travel planning is in some sense a much more "natural" domain and at least partially able to deal with this kind of criticism. We first review the main existing scenarios and paradigms like Lohhausen, Tailorshop, and Moro and compare them to what we call the TRAVELPLAN problem. This problem contains a number of computationally well-investigated problems, which are worked out and can be described by so-called constrained satisfaction problems. The formal investigations have led to the development of a computational architecture which is able to deal with these kinds of subproblems simultaneously. More important, however, is that it serves as a basis for developing experiments and particularly to determine aspects of the computational complexity of the main problem. This in turn allows us to specify and to formulate experimental ideas. Finally, the status quo of ongoing experiments is briefly presented. PMID- 19902285 TI - Diploidy and the selective advantage for sexual reproduction in unicellular organisms. AB - This article develops mathematical models describing the evolutionary dynamics of both asexually and sexually reproducing populations of diploid unicellular organisms. The asexual and sexual life cycles are based on the asexual and sexual life cycles in Saccharomyces cerevisiae, Baker's yeast, which normally reproduces by asexual budding, but switches to sexual reproduction when stressed. The mathematical models consider three reproduction pathways: (1) Asexual reproduction, (2) self-fertilization, and (3) sexual reproduction. We also consider two forms of genome organization. In the first case, we assume that the genome consists of two multi-gene chromosomes, whereas in the second case, we consider the opposite extreme and assume that each gene defines a separate chromosome, which we call the multi-chromosome genome. These two cases are considered to explore the role that recombination has on the mutation-selection balance and the selective advantage of the various reproduction strategies. We assume that the purpose of diploidy is to provide redundancy, so that damage to a gene may be repaired using the other, presumably undamaged copy (a process known as homologous recombination repair). As a result, we assume that the fitness of the organism only depends on the number of homologous gene pairs that contain at least one functional copy of a given gene. If the organism has at least one functional copy of every gene in the genome, we assume a fitness of 1. In general, if the organism has l homologous pairs that lack a functional copy of the given gene, then the fitness of the organism is kappa(l). The kappa(l) are assumed to be monotonically decreasing, so that kappa(0) = 1 > kappa(1) > kappa(2) > cdots, three dots, centered > kappa(infinity) = 0. For nearly all of the reproduction strategies we consider, we find, in the limit of large N, that the mean fitness at mutation-selection balance is max{2e(-mu) - 1,0} where N is the number of genes in the haploid set of the genome, epsilon is the probability that a given DNA template strand of a given gene produces a mutated daughter during replication, and mu = Nepsilon. The only exception is the sexual reproduction pathway for the multi-chromosomed genome. Assuming a multiplicative fitness landscape where kappa(l) = alpha(l) for alpha in (0, 1), this strategy is found to have a mean fitness that exceeds the mean fitness of all the other strategies. Furthermore, while other reproduction strategies experience a total loss of viability due to the steady accumulation of deleterious mutations once mu exceeds [Formula: see text] no such transition occurs in the sexual pathway. Indeed, in the limit as alpha --> 1 for the multiplicative landscape, we can show that the mean fitness for the sexual pathway with the multi-chromosomed genome converges to e(-2mu), which is always positive. We explicitly allow for mitotic recombination in this study, which, in contrast to previous studies using different models, does not have any advantage over other asexual reproduction strategies. The results of this article provide a basis for understanding the selective advantage of the specific meiotic pathway that is employed by sexually reproducing organisms. The results of this article also suggest an explanation for why unicellular organisms such as Saccharomyces cerevisiae (Baker's yeast) switch to a sexual mode of reproduction when stressed. While the results of this article are based on modeling mutation-propagation in unicellular organisms, they nevertheless suggest that, in more complex organisms with significantly larger genomes, sex is necessary to prevent the loss of viability of a population due to genetic drift. Finally, and perhaps most importantly, the results of this article demonstrate a selective advantage for sexual reproduction with fewer and much less restrictive assumptions than those of previous studies. PMID- 19902286 TI - [Think first then manage. Upper gastro-intestinal tract - neoadjuvant therapy]. PMID- 19902287 TI - [Neoadjuvant therapy in the upper gastro-intestinal tract. Modern strategies for Barrett's cancer]. AB - While primary surgical resection with systematic lymphadenectomy remains the treatment of choice for locoregional Barrett's cancer, neoadjuvant chemotherapy is an increasingly accepted treatment modality for patients with locally advanced tumors and patients with extensive lymphatic spread. In contrast to neoadjuvant radiochemotherapy preoperative chemotherapy alone does not seem to increase peri operative complications and mortality. Responders to pre-operative treatment clearly have a survival advantage as compared to those who do not respond. The use of positron emission tomography to measure changes in glucose metabolism of the primary tumor can predict response early after initiation of neoadjuvant chemotherapy and thus help to select patients who will or will not benefit from this approach. The best treatment strategy for non-responders to neoadjuvant therapy remains to be defined. PMID- 19902288 TI - [Therapy in gastric cancer. From an oncological perspective]. AB - In two randomized trials it was demonstrated that preoperative and postoperative chemotherapy shows a statistically significant and clinically relevant improvement in progression-free and overall survival for adenocarcinoma of the esophagogastric junction and stomach when compared with the surgical control arm. The absolute benefit in overall survival was 13% and 14% after 5 years. This benefit is clearly shown for patients with locally advanced tumors but remains debatable in early disease stages. Postoperative mortality and the complication rate were not increased. Based on published study results perioperative chemotherapy has to be regarded as the new standard of care at least for patients staged as uT3/uT4 tumors as defined by endoscopic ultrasound. To date there are no indications for adjuvant chemotherapy or chemoradiation after R0 resection and adequate surgery. PMID- 19902293 TI - The relationship between alcohol intake and cellular immune activity in systemic lupus erythematosus may change from inhibitory to stimulatory within 2 months of study: findings from an integrative single-case study. PMID- 19902289 TI - [Surgery of thyroid carcinoma]. AB - The 5 main types of thyroid cancer (papillary, PTC, follicular, FTC, poorly differentiated, PDTC undifferentiated, UTC, medullary, MTC) not only differ regarding morphology, pathogenesis, genetics,and pathophysiology (iodine metabolism, thyroglobulin and calcitonin production), but also concerning tumor biology, metastatic behavior (lymphogenous, locally invasive and hematogenous routes) and prognosis. Knowledge of these features is the basis of the surgical concept of one or two-stage thyroidectomy, the exceptions and the concept of locoregional lymph node dissection. Lymph node surgery plays an important role in those cancers exhibiting mainly lymph node metastases (PTC, MTC) not only due to frequent recurrences but also due to its potential curative intent. Differentiated carcinomas may have an acceptable prognosis despite local invasion of the cervical aerodigestive system, thus resections are justified when technical prerequisites are given. PMID- 19902294 TI - Three paradoxes of habitat conservation plans. AB - Habitat conservation plans (HCPs) are enabled under section 10(a) of the Endangered Species Act. The substantial increase since 1994 in the number of HCPs has motivated numerous critiques of nearly every aspect of HCPs. These critiques have overlooked several paradoxes that expose fundamental shortcomings of section 10(a) or its implementation. I refer to them as: the Trainwreck Paradox, the Jeopardy Paradox, and the Maximum Mitigation Paradox. The Trainwreck Paradox states that HCPs are needed to avert the listing of species as threatened or endangered, but federal listings are needed to motivate landowners to develop HCPs. The Jeopardy Paradox stems from the vague language of section 10(a) which allows an HCP to reduce the likelihood of a species' survival and recovery but establishes no objective limit on the magnitude of reduction. The Maximum Mitigation Paradox argues that if a landowner provides maximum mitigation at the onset of an HCP, then there will be no financial resources for adaptive management in the future, but if resources are reserved for adaptive management, then the landowner is not mitigating to the maximum extent practicable as required by section 10(a). The purpose of this article is to explain these paradoxes of HCPs and discuss potential remedies. PMID- 19902295 TI - Effects of information on the social acceptability of alternatives to clearfelling in Australian wet eucalypt forests. AB - The effects of viewing different types of information were investigated in people judging the social acceptability of alternative forest harvest systems. Approximately 500 Tasmanians were shown still-simulated images of four harvest systems (a clearfell system, two aggregated retention systems, and a selective system) and were asked to judge their acceptability. Individual interviews were conducted with 12 of the participants. It was anticipated that people holding different beliefs about the consequences of harvesting would have different responses to information. Cluster analysis was used to group participants according to these beliefs. Responses to still images were compared with responses to two other types of information: information about consequences of the harvest systems in the form of indicator symbols, and information about regeneration over time, presented as visual animations. The effects of information differed across both harvest system and belief cluster groups of participants. The largest effects of information occurred in people who held a mix of beliefs about consequences. Within this group, participants who viewed the indicators rated a 30% aggregated retention system higher and selective harvesting lower, than those who did not view the indicators. Viewing animated sequences led to slightly higher ratings of the more intensive harvest systems and significantly lower ratings of the selective harvest system than those based on the still images. The interview data provided examples of interviewees viewing information critically against their own values and beliefs. Only some interviewees appeared to use it in judging social acceptability. PMID- 19902296 TI - Septic complication after balloon-occluded retrograde transvenous obliteration of duodenal variceal bleeding. AB - We report a 64-year-old woman with duodenal varices who underwent balloon occluded retrograde transvenous obliteration (B-RTO) complicated by intraprocedural variceal rupture. The patient developed shivering and a fever higher than 40 degrees C 3 days after the B-RTO procedure. A blood culture grew Entereobacter cloacoe. This case represents a rare septic complication of B-RTO for duodenal varices. PMID- 19902297 TI - Effects of hearing preservation on psychophysical responses to cochlear implant stimulation. AB - Previous studies have shown that residual acoustic hearing supplements cochlear implant function to improve speech recognition in noise as well as perception of music. The current study had two primary objectives. First, we sought to determine how cochlear implantation and electrical stimulation over a time period of 14 to 21 months influence cochlear structures such as hair cells and spiral ganglion neurons. Second, we sought to investigate whether the structures that provide acoustic hearing also affect the perception of electrical stimulation. We compared psychophysical responses to cochlear implant stimulation in two groups of adult guinea pigs. Group I (11 animals) received a cochlear implant in a previously untreated ear, while group II (ten animals) received a cochlear implant in an ear that had been previously infused with neomycin to destroy hearing. Psychophysical thresholds were measured in response to pulse-train and sinusoidal stimuli. Histological analysis of all group I animals and a subset of group II animals was performed. Nine of the 11 group I animals showed survival of the organ of Corti and spiral ganglion neurons adjacent to the electrode array. All group I animals showed survival of these elements in regions apical to the electrode array. Group II animals that were examined histologically showed complete loss of the organ of Corti in regions adjacent and apical to the electrode array and severe spiral ganglion neuron loss, consistent with previous reports for neomycin-treated ears. Behaviorally, group II animals had significantly lower thresholds than group I animals in response to 100 Hz sinusoidal stimuli. However, group I animals had significantly lower thresholds than group II animals in response to pulse-train stimuli (0.02 ms/phase; 156 to 5,000 pps). Additionally, the two groups showed distinct threshold versus pulse rate functions. We hypothesize that the differences in detection thresholds between groups are caused by the electrical activation of the hair cells in group I animals and/or differences between groups in the condition of the spiral ganglion neurons. PMID- 19902298 TI - Evaluation of negation and uncertainty detection and its impact on precision and recall in search. AB - Radiology reports contain information that can be mined using a search engine for teaching, research, and quality assurance purposes. Current search engines look for exact matches to the search term, but they do not differentiate between reports in which the search term appears in a positive context (i.e., being present) from those in which the search term appears in the context of negation and uncertainty. We describe RadReportMiner, a context-aware search engine, and compare its retrieval performance with a generic search engine, Google Desktop. We created a corpus of 464 radiology reports which described at least one of five findings (appendicitis, hydronephrosis, fracture, optic neuritis, and pneumonia). Each report was classified by a radiologist as positive (finding described to be present) or negative (finding described to be absent or uncertain). The same reports were then classified by RadReportMiner and Google Desktop. RadReportMiner achieved a higher precision (81%), compared with Google Desktop (27%; p < 0.0001). RadReportMiner had a lower recall (72%) compared with Google Desktop (87%; p = 0.006). We conclude that adding negation and uncertainty identification to a word-based radiology report search engine improves the precision of search results over a search engine that does not take this information into account. Our approach may be useful to adopt into current report retrieval systems to help radiologists to more accurately search for radiology reports. PMID- 19902299 TI - Evaluating eyegaze targeting to improve mouse pointing for radiology tasks. AB - In current radiologists' workstations, a scroll mouse is typically used as the primary input device for navigating image slices and conducting operations on an image. Radiological analysis and diagnosis rely on careful observation and annotation of medical images. During analysis of 3D MRI and CT volumes, thousands of mouse clicks are performed everyday, which can cause wrist fatigue. This paper presents a dynamic control-to-display (C-D) gain mouse movement method, controlled by an eyegaze tracker as the target predictor. By adjusting the C-D gain according to the distance to the target, the mouse click targeting time is reduced. Our theoretical and experimental studies show that the mouse movement time to a known target can be reduced by up to 15%. We also present an experiment with 12 participants to evaluate the role of eyegaze targeting in the realistic situation of unknown target positions. These results indicate that using eyegaze to predict the target position, the dynamic C-D gain method can improve pointing performance by 8% and reduce the error rate over traditional mouse movement. PMID- 19902300 TI - Novel computer-aided diagnosis algorithms on ultrasound image: effects on solid breast masses discrimination. AB - The objective of this study is to retrospectively investigate whether using the newly developed algorithms would improve radiologists' accuracy for discriminating malignant masses from benign ones on ultrasonographic (US) images. Five radiologists blinded to the histological results and clinical history independently interpreted 226 cases according to the sonographic lexicon of the fourth edition of the Breast Imaging Reporting and Data System and assigned a final assessment category to indicate the probability of malignancy. For each case, each radiologist provided three diagnoses: first with the original images, subsequently with the assistant of the resulting images processed by the proposed CAD algorithms which are called as processed images, and another using the processed images only. Observers' malignancy rating data were analyzed with the receiver operating characteristic (ROC) curve. For reading only with the processed images, areas under the ROC curve (A(z)) of each reader (0.863, 0.867, 0.859, 0.868, 0.878) were better than that with the original images (0.772, 0.807, 0.796, 0.828, 0.846), difference of the average A(z) between the twice reading was significant (p < 0.001). Compared with the results single used processed images, A(z) of utilizing the combined images were increased (0.866, 0.885, 0.872, 0.894, 0.903), but the difference is not statistically significant (p = 0.081). The proposed CAD method has potential to be a good aid to radiologists in distinguishing malignant breast solid masses from benign ones. PMID- 19902301 TI - Influence of catastrophic climatic events and human waste on Vibrio distribution in the Karnaphuli estuary, Bangladesh. AB - Vibrios are bacteria of marine and estuarine origin that can cause human diseases, such as cholera, and also affect aquatic organisms. The impact of storm driven changes in salinity and suspended particulate matter (SPM) on cultivable Vibrio counts (CVC) and distribution in Karnaphuli estuary, Bangladesh, was compared before and after a strong cyclone in mid May 2007 and after a monsoon landslide a month later. CVC were higher (approximately 10(3) colony forming units-cfu/ml) at estuary's mouth (salinity 20-15 parts per thousand, ppt) and steeply declined landwards. CVC and their proportion of total aerobic bacteria were highest after the cyclone and also increased after the landslide, likely due to higher SPM loads. The cyclone did not significantly change previous fecal coliform abundance, contrasting with the ten times increase after the landslide. Sewage input enhanced CVC near the point sources. CVC and salinity correlated highly significantly at salinities <10 ppt; however, at higher values dispersion increased, probably due to the effect of sediment resuspension on CVC. Cyclone or heavy rainfall-mediated turbidity changes jointly with salinity gradients can significantly influence abundance and distribution of estuarine vibrios. Extended salt intrusion and higher turbidities in tropical estuaries by stronger and more frequent storms and deforestation-derived erosion could favor Vibrio growth, with increasing risks for aquatic resources and human health in the coastal zone. PMID- 19902302 TI - In situ silicone tube microextraction: a new method for undisturbed sampling of root-exuded thiophenes from marigold (Tagetes erecta L.) in soil. AB - The difficulties of monitoring allelochemical concentrations in soil and their dynamics over time have been a major barrier to testing hypotheses of allelopathic effects. Here, we evaluate three diffusive sampling strategies that employ polydimethylsiloxane (PDMS) sorbents to map the spatial distribution and temporal dynamics of root-exuded thiophenes from the African marigold, Tagetes erecta. Solid phase root zone extraction (SPRE) probes constructed by inserting stainless steel wire into PDMS tubing were used to monitor thiophene concentrations at various depths beneath marigolds growing in PVC pipes. PDMS sheets were used to map the distribution of thiophenes beneath marigolds grown in thin glass boxes. Concentrations of the two major marigold thiophenes measured by these two methods were extremely variable in both space and time. Dissection and analysis of roots indicated that distribution of thiophenes in marigold roots also was quite variable. A third approach used 1 m lengths of PDMS microtubing placed in marigold soil for repeated sampling of soil without disturbance of the roots. The two ends of the tubing remained out of the soil so that solvent could be washed through the tubing to collect samples for HPLC analysis. Unlike the other two methods, initial experiments with this approach show more uniformity of response, and suggest that soil concentrations of marigold thiophenes are affected greatly even by minimal disturbance of the soil. Silicone tube microextraction gave a linear response for alpha-terthienyl when maintained in soils spiked with 0-10 ppm of this thiophene. This method, which is experimentally simple and uses inexpensive materials, should be broadly applicable to the measurement of non-polar root exudates, and thus provides a means to test hypotheses about the role of root exudates in plant-plant and other interactions. PMID- 19902303 TI - Behavioral and electrophysiological responses of Triatoma brasiliensis males to volatiles produced in the metasternal glands of females. AB - In many insects, mate finding is mediated by volatile sex pheromones, but evidence for this phenomenon in triatomines (Heteroptera: Reduviidae) is still fragmentary. Recently, it was shown that metasternal glands (MGs) are involved in producing signals related to the sexual communication of Triatoma infestans and Rhodnius prolixus. Based on this, we tested whether MG volatiles could be involved in the sexual communication of Triatoma brasiliensis. Odor-mediated orientation responses were studied by using a T-tube olfactometer. These tests showed that males exhibit positive anemotaxis when confronted with adult odor laden air currents. Moreover, females that had their metasternal glands occluded did not elicit significant orientation by males. Compounds produced by the MGs of T. brasiliensis females were identified by means of SPME, GC-FID, and GC-MS, with achiral and chiral columns. All substances identified were ketones and alcohols, and similar compound profiles were found in the secretions produced by both sexes. The most abundant compounds identified were 3-pentanone, followed by (4R) methyl-1-heptanol, 3-pentanol, and (2S)-methyl-1-butanol. In addition, GC-EAD recordings showed that the antennae of males responded to several of the main components of female MG secretions. Our results showed that compounds produced by the MGs of T. brasiliensis females are involved in the sexual communication of this species. PMID- 19902304 TI - Volatiles from a mite-infested spruce clone and their effects on pine weevil behavior. AB - Induced responses by Norway spruce (Picea abies) seedlings to feeding damage by two mite species were studied by analyzing the volatiles emitted during infestation. Four specimens of a Norway spruce (Picea abies L.) clone were infested with mites of Nalepella sp., another four with Oligonychus ununguis, and four were kept mite-free as controls. After a year of infestation, spruce volatiles were collected, analyzed, and identified using SPME-GC-MS. In addition, enantiomers of chiral limonene and linalool were separated by two-dimensional GC. Methyl salicylate (MeSA), (-)-linalool, (E)-beta-farnesene, and (E,E)-alpha farnesene were the main volatiles induced by both species of mites, albeit in different proportions. The ability of the main compounds emitted by the mite infested spruces to attract or repel the pine weevil, Hylobius abietis (L.), was tested. (E)-beta-farnesene was found to be attractive in the absence of spruce odor, whereas methyl salicylate had a deterrent effect in combination with attractive spruce odor. The other tested compounds had no significant effects on the behavior of the weevils. PMID- 19902305 TI - Spatial displacement of release point can enhance activity of an attractant pheromone synergist of a bark beetle. AB - Flight responses of the southern pine beetle, Dendroctonus frontalis Zimmermann, to widely-spaced (>130 m) traps baited with pine volatiles (in turpentine) and the female-produced pheromone component frontalin were enhanced when a bait containing the male pheromone component (+)-endo-brevicomin was attached directly to the trap. However, displacing this bait 4-16 m horizontally from the trap significantly increased its synergistic effect. (+)-endo-Brevicomin enhanced catch to the same degree when the bait was positioned either on the trap or 32 m away. In another experiment, pairs of frontalin/turpentine-baited traps were established with 4 m spacing between traps and >100 m spacing between pairs. Attachment of either a racemic or (+)-endo-brevicomin bait to one trap of a pair caused a significant increase in catch by both traps, but catch in the trap lacking endo-brevicomin was increased more than in its endo-brevicomin-baited twin. In a third experiment, widely-spaced groups of three traps (in a line with 1 and 4 m spacing between the middle and outer traps) were baited uniformly with frontalin and turpentine, and the release rate of (+)-endo-brevicomin from the middle trap was varied across three orders of magnitude. Release rates sufficient to enhance total D. frontalis catch by the trio also caused relatively higher catches to occur in the outer traps than in the middle one. These experiments indicated that both male and female D. frontalis fly to and land preferentially at sources of frontalin and host odors when these are located some distance away from a source of endo-brevicomin. This behavior may have evolved in D. frontalis to allow host-seeking beetles to locate growing, multi-tree infestations while avoiding fully-colonized trees within these infestations. Our data demonstrate that trap spacing alone can qualitatively change the outcome of bait evaluation trials and may explain why many earlier experiments with endo-brevicomin failed to identify it as an aggregation pheromone synergist for D. frontalis. We believe that important aggregative functions of semiochemicals of other bark beetle species may have been similarly overlooked due to choice of experimental procedures. PMID- 19902306 TI - Primary malignant giant-cell tumor of bone has high survival rate. AB - BACKGROUND: Malignant giant-cell tumors (MGCT) comprise 2-9% of all giant-cell tumors (GCTs). The limited existing studies fail to distinguish primary, secondary, or postradiation cancers, making it difficult to design rational treatment strategies. This study compared malignant to benign GCTs and defined the clinical outcome of the patients in a large series of case-matched patients from a single institution. METHODS: Clinical, radiological, and outcome features were compared between 26 malignant and 244 benign GCTs treated in our institution. Five postradiation cancers were excluded. We also performed a 1:2 case-matched comparison of patients with malignant and benign disease. RESULTS: Distal femur (P = 0.019), proximal tibia (P = 0.032), and distal tibia (P = 0.049) had a higher frequency of MGCT. Campanacci stage 1 tumors had a low probability of malignancy (P = 0.017). MGCT were less likely to have aneurysmal bone cyst changes. The 5-year recurrence-free status probability was 80% for malignant and 91% for benign cases in matched groups. The difference in the recurrence rate between benign and malignant groups was not statistically significant (P = 0.24). Functional impairment and limited activity were greater in MGCT patients than in benign GCT patients, whether treated by resection/amputation or curettage/cryosurgery. CONCLUSIONS: We found that malignant and benign GCT have similar epidemiology and that recurrence was higher in MGCT (20 v 9%). Local recurrence for MGCT was not statistically different for excision versus intralesional therapy, but there was little statistical power. Finally, the 16% mortality for patients with MGCT suggests low-grade malignancy. PMID- 19902307 TI - Briefings, checklists, geese, and surgical safety. PMID- 19902308 TI - Functional recovery following an end to side neurorrhaphy of the accessory nerve to the suprascapular nerve: case report. AB - The use of end-to-side neurrorhaphy remains a controversial topic in peripheral nerve surgery. The authors report the long-term functional outcome following a modified end-to-side motor reinnervation using the spinal accessory to innervate the suprascapular nerve following a C5 to C6 avulsion injury. Additionally, functional outcomes of an end-to-end neurotization of the triceps branch to the axillary nerve and double fascicular transfer of the ulnar and medial nerve to the biceps and brachialis are presented. Excellent functional recoveries are found in respect to shoulder abduction and flexion and elbow flexion. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11552 009-9242-3) contains supplementary material, which is available to authorized users. PMID- 19902309 TI - The Wide-Awake Approach to Dupuytren's Disease: Fasciectomy under Local Anesthetic with Epinephrine. AB - The Wide-Awake Approach to Dupuytren's contracture involves fasciectomy under local anesthetic with epinephrine and no tourniquet. The goal of this study is to show that the Wide-Awake Approach produces equivalent outcomes to fasciectomy under general anesthetic with a tourniquet, with fewer risks to the patient. A multicenter retrospective review was conducted on 111 patients with fasciectomies under local or general anesthetic between 2001 and 2007. Data on patient demographics, comorbidities, cost, as well as range of motion was collected and evaluated using Microsoft Excel and SAS. Of 148 fingers, 102 were treated under local and 46 under general anesthetic. The average postoperative Total Active Motion (TAM) for general anesthetic patients was 199.0 +/- 29.6 (D5), 223.9 +/- 29.3 (D4), 234.6 +/- 14.6 (D3), and 246.7 +/- 14.4 (D2). The average postoperative TAM for local anesthetic patients was 168.3 +/- 62.2 (D5), 195.9 +/ 67.5 (D4), 173.0 +/- 72.6 (D3), and 177.5 +/- 31.8 (D2). There were no significant differences between any of these individual groups (p = 0.09, 0.26, 0.12, and 0.20, respectively); however, when pooled, the overall TAM was significantly greater in the general anesthesia group (222.0 +/- 29.7 vs. 186.0 +/- 63.0, p = 0.002.). Complication rates and types were similar with both techniques. The Wide-Awake Approach to Dupuytren's contracture avoids general anesthetic risks and has cost benefits to healthcare providers. Although it yields similar range of motion outcomes to fasciectomy performed under general anesthesia, total active motion may be better with fasciectomy done under general anesthesia. PMID- 19902310 TI - Endoluminal fundoplication (ELF) for GERD using EsophyX: a 12-month follow-up in a single-center experience. AB - BACKGROUND: Several endoscopic antireflux therapies have been proposed to reduce the need for chronic medical therapy or laparoscopic fundoplication for gastroesophageal reflux disease (GERD). Aim of this study was to evaluate the short- and mid-term clinical results of endoluminal fundoplication (ELF) with EsophyX. PATIENTS AND METHODS: From June 2006 to April 2008, 20 patients were enrolled in the study. All the ELFs were performed under general anesthesia. RESULTS: The mean duration of the procedure was 63 min (range 38-105). A median of 14 fasteners was placed. There were no major intraoperative complications. Two patients developed early complications and were treated conservatively. Four patients underwent, within the first year post-ELF, a laparoscopic fundoplication because of persistence of symptoms. One patient was lost to follow-up between 6 and 12 months. Among the other 15 patients who completed 12 months follow-up, the GERD health-related quality of life score decreased from a median of 40 to a median of 10 (p < 0.05), and seven patients were still off proton pump inhibitor. An improvement in esophageal acid exposure was recorded in 16.6% of patients, while in 66.7%, it worsened. CONCLUSIONS: ELF induced improvement of GERD symptoms and patients quality of life in a subgroup of patients with a reduced need for medication. However, it did not significantly change esophageal acid exposure in these patients. The need for revisional standard laparoscopic fundoplication was high. PMID- 19902311 TI - Surgical management of infrahilar/suprapancreatic cholangiocarcinoma: an analysis of the surgical procedures, surgical margins, and survivals of 77 patients. AB - BACKGROUND: Optical surgical management of infrahilar/suprapancreatic cholangiocarcinoma remains controversial. METHODS: Between 1988 and 2006, 77 patients with infrahilar/suprapancreatic cholangiocarcinoma underwent curative surgical resections following our intention-to-treat strategy. The clinicopathological factors affecting survival were evaluated using univariate and multivariate analyses with regard to the surgical procedures and surgical margins. RESULTS: The surgical procedure included extrahepatic bile duct resection alone (EHBD; n = 17), major hepatectomy combined with extrahepatic bile duct resection (MHx; n = 26), pancreaticoduodenectomy (PD; n = 28), and MHx and concomitant PD (HPD; n = 6). Performance of MHx and/or PD in addition to EHBD increased surgical morbidity (p = 0.001). Among patients undergoing the four surgical procedures (EHBD, MHx, PD, and HPD), no significant difference was found in the incidence of positive overall surgical margins (53%, 65%, 46%, and 67%, p = 0.51) or long-term survivals (median survival time, 51, 27, 41, and 22 months, p = 0.60). A multivariate analysis revealed that perineural invasion (95% confidence interval, 1.1-12.3, p = 0.009), nodal metastasis (1.6-6.8, p = 0.001), and blood transfusion (1.1-3.9, p = 0.02) were independent predictors of a poor outcome. Perineural invasion was associated with positive radial margins (p = 0.045) and submucosal ductal infiltration (p = 0.03). CONCLUSION: Perineural invasion, rather than the type of surgical procedure, had a significant impact on surgical curability and survival of patients with infrahilar/suprapancreatic cholangiocarcinoma treated according to our intention-to-treat strategy. PMID- 19902312 TI - Detection of free peritoneal cancer cells in gastric cancer using cancer-specific Newcastle disease virus. AB - INTRODUCTION: Cytologic detection of peritoneal gastric cancer cells by Papanicolaou staining offers important prognostic information but has low sensitivity. We evaluated a novel detection technique using Newcastle disease virus expressing the enhanced green fluorescent protein (NDV-GFP) gene. METHODS: NDV-GFP was tested on MKN-1 human gastric adenocarcinoma cells plated upon rat hepatocytes to determine tumor-specific infection and GFP expression. Malignant ascites infected with increasing doses of virus was evaluated for NDV-GFP dose determination. Peritoneal lavage samples from 30 patients with gastric adenocarcinoma undergoing staging laparoscopy were evaluated with NDV-GFP. RESULTS: NDV-GFP can specifically detect one MKN-1 cell among one million benign rat hepatocytes. NDV-GFP at 5 x 10(6) plaque-forming units (PFU) produced optimal GFP expression in malignant ascites. Noncancerous cells were non-GFP expressing. GFP-expressing cells counterstained positive for carcinoembryonic antigen expression, confirming their cancerous origin. Furthermore, in patients with advanced gastric cancer, GFP expression was markedly enhanced over cytology. Of six patients with M1 disease discovered during laparoscopy, only 50% were cytology positive. All six, however, were NDV-GFP positive. Cytology was positive in 9% of patients with T3 disease, 8% with N1 disease, and 50% with N2 disease. In contrast, NDV-GFP was positive in 95% of T3 patients and 100% of patients with N1 or N2 disease. CONCLUSIONS: NDV-GFP can specifically infect and detect peritoneal gastric cancer cells and offers a more sensitive method compared with conventional cytology. This novel modality may offer enhanced detection of intraperitoneal cancer spread and provide important prognostic information. PMID- 19902313 TI - Subclinical intestinal inflammation in patients with Crohn's disease following bowel resection: a smoldering fire. AB - BACKGROUND/AIMS: Fecal lactoferrin is the direct expression of intestinal inflammation in Crohn's disease (CD). The aim of this study was to analyze the in vivo intimate correlation between intestinal and systemic inflammation in CD patients in clinical remission following bowel resection. The secondary end point was to evaluate the prognostic value of lactoferrin levels and serum cytokines in terms of need of surgery for recurrence in these patients. PATIENTS AND METHODS: Fecal lactoferrin and serum cytokine (interleukin (IL)-1beta, IL-6, IL-12, tumor necrosis factor (TNF)-alpha, and transforming growth factor (TGF)-beta1) levels were assessed; hematological and biochemical investigations were carried out, and Crohn's Disease Activity Index was evaluated in the 36 patients who had undergone bowel resection. The prognostic value of lactoferrin and cytokine levels in terms of surgical recurrence was assessed by re-calling patients after 24 months from the enrolment in the study. RESULTS: All patients, evaluated after a follow-up of 36 +/- 5 months, were in clinical remission. Fecal lactoferrin levels were found to be significantly correlated with IL-6 (R = 0.431, p = 0.025) and C-reactive protein (CRP; R = 0.507, p = 0.007), while no correlation was observed between lactoferrin and IL-1beta, IL-12, TNF-alpha, or TGF-beta1. Reoperation for anastomotic recurrence tended to occur significantly more frequently in patients with higher IL-6 (p = 0.10). CONCLUSIONS: Subclinical intestinal inflammation, expressed by fecal lactoferrin, seems to keep the systemic inflammation alive in CD patients through the IL-6-CRP cascade. IL-6 seems to be predictive of the outcome of CD patients undergoing surgery. PMID- 19902314 TI - Effectiveness of prophylactic antibiotics in a population-based cohort of patients undergoing planned cholecystectomy. AB - BACKGROUND: In the absence of randomized controlled trials with sufficient power to assess the effectiveness of prophylactic antibiotics (PA), the best evidence is provided by large population-based register studies. METHODS: The Swedish Register of Gallstone Surgery and ERCP (GallRiks) started in May 2005 and reached 75% national coverage in 2007. During 2006 and 2007, a total of 16,400 operations were registered in GallRiks. In the present study, all elective procedures performed in 2006-2007 in units performing at least 25 operations annually were included in an analysis of the risk for postoperative infectious complications RESULTS: Altogether 10,927 procedures were performed 2006-2007. Univariate logistic regression analysis revealed a paradoxical increase in postoperative infectious complications requiring antibiotic treatment and postoperative abscess if PA were given (p < 0.05). This increase disappeared in multivariate analysis with adjustment for age, gender, presence of cholecystitis, accidental gallbladder perforation, and presence of bile duct stones. CONCLUSION: No benefit from PA was seen in this study on elective cholecystectomy. Although a randomized controlled trial could possibly show a reduction in the risk for postoperative infectious complications not detected in this study, such a reduction must be weighed against the risk of promoting drug resistance by the widespread use of PA. PMID- 19902315 TI - Preoperative weight loss: a component of the preoperative program in bariatric surgery. PMID- 19902316 TI - Two-year results on morbidity, weight loss and quality of life of sleeve gastrectomy as first procedure, sleeve gastrectomy after failure of gastric banding and gastric banding. AB - BACKGROUND: Sleeve gastrectomy (SG) is an alternative to gastric bypass and laparoscopic adjustable gastric banding (GB). METHODS: From January 2004 to January 2006, 111 patients with a follow-up longer than 24 months were prospectively followed. Three treatment groups were defined. Sleeve gastrectomy as first procedure (SGFP; n = 50), sleeve gastrectomy after failure of GB (SG after GB; n = 9) and GB (n = 52). We compared morbidity, mortality, length of stay, number of procedures under general anaesthesia, excess weight loss (EWL) and quality of life. RESULTS: Mean initial body mass index (BMI) was 50.4 (SG), 50.8 (SG after GB) and 43.8 (GB; p = 0.000001). Mean operating time was 97.1 min (SGFP), 122.2 min (SG after GB) and 69.8 min (GB; p < 0.0001). The reoperation rate under general anaesthesia was 2% (SGFP), 11% (SG after GB) and 30.76% (GB; p = 0.00001).The fistula rate was 2% (SGFP), 0% (SG after GB) and 0% (GB). BMI at 24 months was 33.8 (SGFP), 35.3 (SG after GB) and 33.2 (GB; NS). EWL at 24 months was 67.4 (SGFP), 60.3 (SG after GB) and 58.6 (GB; NS). In the SGFP group and in the SG after GB group, the mean quality-of-life score was 1.1. In the GB group, the mean score was 0.95 (NS). CONCLUSIONS: Initial BMI was significantly higher in the SG group but was no longer significantly different from the BMI of the GB group at 12 and 24 months. Excess BMI loss was higher after SG than after GB. This reduction of BMI was considered to be a success for GB. Thus, results of SG should be considered as a success. Quality of life was not significantly different between the three groups. These results validated SG as first procedure or after failure of GB. PMID- 19902317 TI - Alterations of central dopamine receptors before and after gastric bypass surgery. AB - BACKGROUND: While bariatric surgery has proved highly successful at producing sustained weight loss, variability in treatment response persists. A better understanding of the pathophysiology of appetite and obesity may improve patient selection and management. Research into feeding behavior and satiety has focused on the role of dopamine in reward-based behaviors. Specifically, positron emission computed tomography (PET) has demonstrated reduced brain dopamine receptor availability in obese subjects compared to controls. This may be due to a primary deficiency in dopamine receptors or to secondary dopamine receptor downregulation. We performed a preliminary study to investigate dopamine D2 receptor activity in obese subjects before and after laparoscopic Roux-en Y gastric bypass (LGBP). METHODS: Five female subjects, ages 20 to 38 years old with a mean body mass index of 45, underwent PET with [C-11] raclopride injection. Five regions of interest were studied: ventral striatum, anterior and posterior putamen, and anterior and posterior caudate nucleus. Repeat PET was performed at 6 weeks following LGBP. D2 receptor binding was compared within subjects pre- and post-surgery. Baseline D2 binding was also compared to historical nonobese controls. RESULTS: D2 receptor availability increased 6 weeks after gastric bypass surgery. The increase in receptor availability appeared roughly proportional to the amount of weight lost. No significant difference in D2 binding was seen between the obese subjects and historical nonobese controls. CONCLUSIONS: Brain available dopamine D2 binding appears to increase following GBP. This preliminary finding needs to be replicated in a larger population but suggests that diminished D2 binding in the obese may be due to D2 receptor downregulation. Changes in available dopamine receptor binding may play an important role in centrally mediated appetite suppression and resultant weight loss after LGBP. PMID- 19902318 TI - The fate of spontaneous synchronous rhythms on the cerebrocerebellar loop. AB - How does the cerebellum participate in neocortical rhythms? Neocortical signals destined for the cerebellum are integrated in the pontine nuclei (PN) with cerebellar output signals via a direct, reciprocal feedback loop with the cerebellar nuclei (CN). The present study investigated the fate of two spontaneously occurring rhythms in rat neocortex under ketamine anesthesia-slow wave activity at around 1 Hz and gamma oscillations-within this pontonuclear feedback loop. Coordinated oscillatory neuronal activity was studied using simultaneous multineuron recordings in primary motor cortex (M1), PN, and lateral CN. It was revealed that slow burst firing-known in neocortex as "up and down states"-is readily conveyed within the pontonuclear feedback loop and thus engages the entire cerebropontocerebellothalamic loop. In contrast, gamma band synchronous oscillations reached only the PN under the present experimental conditions. Surprisingly, many CN single units were actually found to oscillate in the gamma range, but they completely failed to synchronize with other units in either CN or PN. These results show firstly that slow concerted activity can readily engage the entire cerebrocerebellar loop. Secondly, they raise the possibility that fast gamma oscillations may be incompatible with cerebellar processing and get blocked out. Future studies in behaving animals are needed to answer the question whether signals coded in gamma band frequency are converted to another carrier code using the feedback control exerted by the pontonuclear loop. PMID- 19902319 TI - 11C-meta-hydroxyephedrine defects persist despite functional improvement in hibernating myocardium. AB - BACKGROUND: Regional cardiac sympathetic nerve dysfunction develops in hibernating myocardium and may play a role in its association with sudden cardiac death. Interventions to improve cardiac function (i.e., revascularization) improve survival, but the potential reversibility of sympathetic nerve dysfunction remains unclear. METHODS AND RESULTS: Pigs (n = 11) were chronically instrumented with a proximal left anterior descending coronary artery (LAD) stenosis to produce hibernating myocardium. Prior to therapeutic interventions, there was LAD occlusion with collateral-dependent myocardium, reduced regional function (echocardiographic LAD wall-thickening 23% +/- 4% vs 83% +/- 6% in Remote, P < .001), and large defects in (11)C-meta-hydroxyephedrine (HED) PET (48% +/- 4% of LV area, 26% +/- 2% integrated reduction). Successful PCI or pravastatin therapy improved regional (LAD wall-thickening 23% +/- 4% to 42% +/- 6%, P < .05) and global LV function (fractional shortening 24% +/- 2% to 31% +/- 2%, P < .01), but did not alter regional HED uptake, retention, defect size, or defect severity. CONCLUSIONS: Despite significant functional improvement of hibernating myocardium as a result of PCI or pravastatin therapy, there were no changes in HED defect size or severity. Thus, inhomogeneity in myocardial sympathetic innervation persisted, and the lack of plasticity suggests that even in the absence of significant infarction, structural rather than functional defects are responsible for reduced myocardial norepinephrine uptake in chronic ischemic heart disease. PMID- 19902320 TI - Peroxisome proliferator-activated receptor-gamma activation attenuates cardiac fibrosis in type 2 diabetic rats: the effect of rosiglitazone on myocardial expression of receptor for advanced glycation end products and of connective tissue growth factor. AB - Receptor for advanced glycation end products (RAGE) and connective tissue growth factor (CTGF) play a key role in diabetic myocardial fibrosis, and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) activation has been reported to reduce RAGE and CTGF expression. This study investigated the effects of the PPAR-gamma agonist, rosiglitazone, on myocardial expression of RAGE and CTGF, extent of cardiac fibrosis, and left ventricular (LV) diastolic function in type 2 diabetic (T2D) rats. Twenty-week-old T2D rats were randomized to treatment with either 20 weeks of rosiglitazone (20 mg/kg) or saline (n = 10 in each group). Serial echocardiographic examinations were performed just before randomization (20 weeks) and at study completion (40 weeks). Fibrosis extent and RAGE and CTGF expression were assessed in previously imaged hearts by picrosirius red staining, and by real-time reverse transcriptase-coupled polymerase chain reaction (RT-PCR) and immunoblotting, respectively. Results of the latter assessments were further validated by immunohistochemical staining. Rosiglitazone treatment significantly improved E/A ratio in serial echocardiography assessment, and reduced LV collagen volume fraction as demonstrated by picrosirius red staining. Real-time RT-PCR and immunoblots of myocardial tissue from rosiglitazone-treated rats revealed reduced RAGE and CTGF mRNA and protein signals compared to those of saline-treated T2D rats, which were consistent with reduced proportions of myocardial RAGE and CTGF staining in the hearts of T2D rats. PPAR-gamma agonist therapy reduces cardiac fibrosis and improves LV diastolic dysfunction as assessed by serial echocardiographic imaging. Suppression of RAGE and CTGF expression in the diabetic myocardium appears to contribute to the antifibrotic effect of rosiglitazone. These results support the potential of PPAR-gamma agonists as antifibrotic agents in diabetic cardiomyopathy. PMID- 19902321 TI - Insights into the salt tolerance mechanism in barley (Hordeum vulgare) from comparisons of cultivars that differ in salt sensitivity. AB - Although barley (Hordeum vulgare L.) is a salt-tolerant crop, the underlying physiological and molecular mechanisms of salt tolerance remain to be elucidated. Therefore, we investigated the response of salt-tolerant (K305) and salt sensitive (I743) cultivars to salt stress at both physiological and molecular levels. Salt treatment increased xylem sap osmolarity, which was attributed primarily to a rise in Na(+) and Cl(-) concentration; enhanced accumulation of the ions in shoots; and reduced plant growth more severely in I743 than K305. The concentration of K(+) in roots and shoots decreased during 8 h of salt treatment in both cultivars but with no marked difference between cultivars. Hence, the severe growth reduction in I743 is attributed to the elevated levels of (mainly) Na(+) in shoots. Analysis of gene expression using quantitative RT-PCR showed that transcripts of K(+)-transporters (HvHAK1 and HvAKT1), vacuolar H(+)-ATPase and inorganic pyrophosphatase (HvHVA/68 and HvHVP1) were more abundant in shoots of K305 than in shoots of I743. Expression of HvHAK1 and Na(+)/H(+) antiporters (HvNHX1, HvNHX3 and HvNHX4) was higher in roots of K305 than in I743 with prolonged exposure to salt. Taken together, these results suggest that the better performance of K305 compared to I743 during salt stress may be related to its greater ability to sequester Na(+) into sub-cellular compartments and/or maintain K(+) homeostasis. PMID- 19902322 TI - The importance of the origin of vertebral arteries in cerebral ischemia in the rabbit. AB - The aim of this study was to verify whether experimentally induced total cerebral ischemia in rabbits actually corresponds to total ischemia on the basis of the origin of certain vessels. We observed morphological variations in the origin and course of the arteria vertebralis as one of the vessels supplying the brain with blood. Investigations were carried out on 50 adult New Zealand rabbits. We prepared corrosion casts of the arterial system using Duracryl Dental. We found that in 86% of cases (43 animals) the arteria vertebralis sinistra originated directly from the arteria subclavia sinistra, in 10% of cases (5 animals) it originated from the arcus aortae as an independent branch, and in 4% of cases (2 animals) it arose from the arcus aortae as a common trunk with the arteria scapularis descendens. The arteria vertebralis dextra originated from the arteria subclavia dextra in 98% (49 animals) of cases. In one case we observed two arteria vertebralis dextra with two different origins. Bilateral variability in the origin of the arteria vertebralis was observed in 12% of cases (6 animals). Our results show that ligation of the truncus brachiocephalicus and of the arteria subclavia sinistra do not necessarily cause total cerebral ischemia. PMID- 19902323 TI - Diadochokinetic movements differ between patients with Parkinson's disease and controls. AB - Instrumental motion assessment, i.e., peg insertion, gains importance as an addition to rating procedures to determine the efficacy of therapeutic interventions and to serve as diagnostic tool in patients with Parkinson's disease (PD). One of their motor features is disturbed execution of alternating motions. The objectives of the study were to assess pronation and supination of the forearms and peg insertion with instruments and to correlate the outcomes with rating scores in PD patients. Controls and 27 idiopathic PD patients, taken off treatment for 12 h, were scored and performed on both devices. PD patients showed a reduced maximum velocity (v), performance interval and amplitude of diadochokinetic movements than controls. V outcomes showed closer associations to rated motor behavior, in particular akinesia and rigidity, and peg insertion results than amplitude, respectively periods of diadochokinetic motion sequences. This altered performance of diadochokinetic movements corresponds to the clinical motor symptoms of PD patients and differs from healthy controls. PMID- 19902324 TI - Association between the RAGE G82S polymorphism and Alzheimer's disease. AB - The receptor for advanced glycation end products (RAGE) is associated with several pathological states including Alzheimer's disease (AD) pathology, while its soluble form (sRAGE) acts as a decoy receptor. We have tested for association of AD with a functional single-nucleotide polymorphism (SNP) in the RAGE gene (G82S; rs2070600), a SNP associated with increased ligand affinity of RAGE. Analysis of a Chinese cohort (276 cases; 254 controls) showed a higher prevalence of the RAGE 82S allele and GS + SS genotype in the patients [82S vs. 82G: P = 0.017, odds ratio (OR) = 1.431; GS + SS vs. GG: P = 0.025, OR = 1.490]. Further stratification analysis revealed that the association of the RAGE G82S polymorphism with AD was significant in early onset AD stratum. Moreover, plasma sRAGE levels were lower in AD than in normal elderly controls, and the presence of the risk allele was associated with further plasma sRAGE reduction and a fast cognitive deterioration. The present study provides preliminary evidence that the RAGE G82S variant is involved in genetic susceptibility to AD. PMID- 19902325 TI - H2O2 localization in the green alga Micrasterias after salt and osmotic stress by TEM-coupled electron energy loss spectroscopy. AB - Reactive oxygen species (ROS), including hydrogen peroxide (H(2)O(2)), are constantly generated as by-products of normal metabolic cellular pathways and can be overproduced in response to stress. In this study, we investigated ROS production and localization of H(2)O(2) after salt (200 mM KCl) and osmotic (iso osmotic sorbitol concentration) stress in the unicellular green alga Micrasterias. By means of the dye H(2)DCFDA and confocal laser scanning microscopy, most ROS production could be detected in KCl-treated cells when compared to sorbitol-exposed cells and controls. For ultrastructural detection of H(2)O(2), CeCl(3), which reacts with H(2)O(2) and produces cerium perhydroxide deposits, has been used. Cerium was identified by transmission electron microscopy (TEM)-coupled electron energy loss spectroscopy (EELS) in organelles of KCl- and sorbitol-treated cells and in controls. Statistical measurements of the presence of the cerium M(4,5) edge were performed in mitochondria, chloroplasts, cell walls, and cytoplasmic sites of five individual cells after each treatment. The most pronounced increase in H(2)O(2) production was found in chloroplasts of KCl- and sorbitol-treated cells. This shows that the chloroplast reveals the strongest response in H(2)O(2) production after stress induction in Micrasterias. Significant elevation of H(2)O(2) production also occurred in mitochondria and cytoplasm, whereas H(2)O(2) levels remained unchanged or even slightly decreased in cell walls of treated cells. Additionally, TEM micrographs and EELS analyses provided indirect evidence for an increased H(2)O(2) production at the plasma membrane of KCl-treated cells, indicating an involvement of the plasma membrane NADPH oxidase in H(2)O(2) generation. PMID- 19902326 TI - Basal levels of DNA damage detected by micronuclei and comet assays in untreated breast cancer patients and healthy women. AB - Breast cancer is the second most frequent type of cancer worldwide and is the most common malignant disease among women. Risk factors for breast cancer include early menarche, late menopause, hormonal therapies, exposure to environmental pollutants, smoking and alcohol use. However, increased or prolonged exposure to estrogen is the most important risk factor. It has been suggested that accumulation of DNA damage may contribute to breast carcinogenesis. Epidemiological studies suggest that cytogenetic biomarkers such as micronuclei in peripheral blood lymphocytes may predict cancer risk because they indicate genomic instability in target tissues. The objective of the present study was to evaluate the frequencies of micronuclei and the extent of DNA damage detected by comet assay in peripheral blood lymphocytes of untreated breast cancer patients and healthy women. The study was conducted using peripheral blood lymphocytes from 45 women diagnosed for Ductal "in situ" or invasive breast carcinoma and 85 healthy control women. Micronuclei and comet assays were performed to detect spontaneous DNA damage. The results showed that micronuclei frequencies and tail intensity, detected by comet assay, were significantly higher in the breast cancer group than in controls. The levels of DNA damage were similar in smokers and non-smokers, and aging did not influence the frequencies of micronuclei or tail intensity values observed in either group. In conclusion, the present work demonstrates higher levels of DNA damage in untreated breast cancer patients than in healthy women. PMID- 19902327 TI - Dynamic magnetic resonance imaging in acute pyelonephritis. AB - PURPOSE: This study was conducted to evaluate the role and clinical impact of dynamic magnetic resonance imaging (MRI) in the diagnosis and follow-up of acute pyelonephritis (APN). MATERIALS AND METHODS: We retrospectively reviewed 442 consecutive renal MRI examinations (279 diagnostic and 163 follow-up) performed in 285 patients (mean age 42.17 years), 35 of whom were kidney transplant recipients with a clinical suspicion of APN. RESULTS: MRI showed signal abnormalities suggestive of APN in 125/244 (51.2%) patients with native kidneys. Except for two examinations performed without paramagnetic contrast material, the inflammatory foci appeared as areas of nonenhancement: single in 39/123 cases, multiple in 84/123, unilateral in 60/84 and bilateral in 24/84. Abscesses were present in 40/123 (32.5%) positive cases. During follow-up, we observed complete normalisation of MRI signs in 86/103 patients; 17/103 (16.5%) cases evolved into fibrosis and scarring. In 15/35 (42.8%) patients with transplanted kidney, MRI was positive for APN. CONCLUSIONS: Renal MRI is an effective tool for the diagnosis and follow-up of APN both in patients not at risk and those at higher risk, such as those with a transplanted kidney. The high costs of the examination are offset by better treatment planning and early complication detection. PMID- 19902328 TI - MR characterisation of dysplastic nodules and hepatocarcinoma in the cirrhotic liver with hepatospecific superparamagnetic contrast agents: pathological correlation in explanted livers. AB - PURPOSE: The authors sought to evaluate the potential of magnetic resonance (MR) imaging with superparamagnetic iron oxide (SPIO) contrast material for identifying, characterising and differentiating dysplastic nodules (DN) and hepatocellular carcinoma (HCC) in the cirrhotic liver by correlating the results with pathological findings on the explanted liver. MATERIAL AND METHODS: MR imaging was used to study the liver of 400 cirrhotic patients awaiting liver transplantation, 31 of whom were selected to receive a liver. Of these 31 patients, we included in the study 22 patients (mean age 53 years; range 46-57 years) who underwent liver transplantation within 12-24 h after MR examination. Patients were studied with a 1.5-T device, and scans were acquired before and after the administration of paramagnetic contrast material. For every lesion, we recorded signal intensity on unenhanced baseline T1- and T2-weighted images and enhancement pattern after SPIO administration. Histological examination of the entire liver provided the definitive diagnosis of the lesions. RESULTS: Histological examination identified 59 lesions: 14 HCC, 4 HCC-DN, 39 DN, and 2 cystoadenomas. Among the 14 HCC, three were well differentiated, eight were moderately differentiated and three were poorly differentiated. Of the 39 DN, 28 were low-grade and 11 high-grade lesions. Unenhanced baseline MR imaging correctly identified and characterised 20 lesions, equal to 33.90% of all lesions: 6 HCC, 12 DN and 2 DN with a subfocus of HCC. SPIO-enhanced MR imaging showed greater sensitivity detecting and characterising 45 lesions, equal to 76.27% of all lesions identified at histology: 14 HCC, 27 DN and 4 DN with subfocus of HCC. SPIO administration improved the sensitivity of MR imaging in lesion detection and characterisation by 42.37%. False negative results with SPIO enhanced MR imaging occurred in 12 DN (31%), which histological examination revealed to be low-grade DN with a diameter <1 cm. CONCLUSIONS: SPIO-enhanced MR imaging proved to be of value in detecting and characterising lesions in the cirrhotic liver, allowing differentiation of DN from HCC and providing an early diagnosis of neoplastic degeneration of DN. PMID- 19902329 TI - Stress-ECG vs. CT coronary angiography for the diagnosis of coronary artery disease: a "real-world" experience. AB - PURPOSE: This study aimed to evaluate the diagnostic accuracy of stress electrocardiogram (ECG) and computed tomography coronary angiography (CTCA) for the detection of significant coronary artery stenosis (> or =50%) in the real world using conventional CA as the reference standard. MATERIALS AND METHODS: A total of 236 consecutive patients (159 men, 77 women; mean age 62.8+/-10.2 years) at moderate risk and with suspected coronary artery disease (CAD) were enrolled in the study and underwent stress ECG, CTCA and CA. The CTCA scan was performed after i.v. administration of a 100-ml bolus of iodinated contrast material. The stress ECG and CTCA reports were used to evaluate diagnostic accuracy compared with CA in the detection of significant stenosis > or =50%. RESULTS: We excluded 16 patients from the analysis because of the nondiagnostic quality of stress ECG and/or CTCA. The prevalence of disease demonstrated at CA was 62% (n=220), 51% in the population with comparable stress ECG and CTCA (n=147) and 84% in the population with equivocal stress ECG (n=73). Stress ECG was classified as equivocal in 73 cases (33.2%), positive in 69 (31.4%) and negative in 78 (35.5%). In the per-patient analysis, the diagnostic accuracy of stress ECG was sensitivity 47%, specificity 53%, positive predictive value (PPV) 51% and negative predictive value (NPV) 49%. On stress ECG, 40 (27.2%) patients were misclassified as negative, and 34 (23.1%) patients with nonsignificant stenosis were overestimated as positive. The diagnostic accuracy of CTCA was sensitivity 96%, specificity 65%, PPV 74% and NPV 94%. CTCA incorrectly classified three (2%) as negative and 25 (17%) as positive. The difference in diagnostic accuracy between stress ECG and CTCA was significant (p<0.01). CONCLUSIONS: CTCA in the real world has significantly higher diagnostic accuracy compared with stress ECG and could be used as a first-line study in patients at moderate risk. PMID- 19902330 TI - Diffusion-weighted imaging in breast lesion evaluation. AB - PURPOSE: The purpose of this study was to investigate the ability of diffusion weighted imaging (DWI) and the apparent diffusion coefficient (ADC) in the detection and characterisation of breast lesions. MATERIALS AND METHODS: From September 2005 to September 2007, 86 patients with breast lesions who underwent magnetic resonance imaging (MRI) in our department were included in our study. MRI was performed with a 1.5-T unit using a standard protocol including DWI sequence. For each breast lesion, the ADC value was calculated and compared with that of normal breast tissue and to the definitive pathological diagnosis. Mann Whitney U and Kruskal-Wallis tests were used for statistical analysis. RESULTS: A total of 126 breast lesions were detected. Pathology results revealed 100 malignant and 26 benign lesions. Mean diameter of lesions was 26.02 mm (range 4 90 mm), including 52 lesions 6, while 8.6% of cases are non organ confined (2 cases and 1 case with extra- capsular and Seminal vesicles infiltrations, respectively). Both lobes were involved in 20 cases. Moreover, 40% (14 cases) showed upgrading of the score. Lymphatic permeation could be recognized in four cases. Mean follow-up time 88.1 months, 8.6% cases develop metastasis. CONCLUSION: Whenever Epstein criteria are applied in Middle East patients, the decision of active surveillance based for clinically insignificant prostate cancer (who met the Epstein criteria) will miss nearly 46% of unfavorable prostate cancer. As well as a risk of later metastasis could be in 8.6% of them. PMID- 19902378 TI - Investigative clinical study on prostate cancer: on the role of the pretreatment total PSA to free testosterone ratio in selecting different biology groups of prostate cancer patients. AB - OBJECTIVES: To show that prostate cancer biology is related to serum levels of both free testosterone (FT) and prostate-specific antigen (PSA), that PSA level is linearly related to FT and that the PSA to FT ratio may be considered as the growth rate parameter expressing cancer phenotype biology. MATERIALS AND METHODS: The study includes 135 consecutive patients diagnosed with prostate cancer. Pretreatment simultaneous serum samples for analyzing total testosterone (TT), FT and total PSA levels were obtained. The study was assessed according to a multidimensional approach of the five continuous variables including TT, FT, PSA, AGE and percentage of positive biopsies (=P+). The all sets of data were considered as one--sample with no groupings among the observations. Multivariate analysis included factor analysis (FA) and principal component analysis (PCA). Multivariate inferential statistics for comparing different groups of patients according to the PSA to free testosterone ratio (PSA/FT) included Hotteling's multivariate two-sample T2-Test for comparing two mean vectors as well as Box's M Test with the chi-square approximation for comparing multiple covariance matrices when patients were sampled in more than two groups. RESULTS: Factor analysis showed the two natural grouping of variables, FT-TT and PSA-P+. PCA assessed FT and PSA as the two variables with large variances having a notable influence on the first two principal components. Multiple linear regression analysis showed that all the income variables, except age, significantly predicted the PSA/FT ratio. Patients were first sampled according to the PSA/FT ratio in group 1 (PSA/FT <= 0.20) and group 2 (PSA/FT > 0.20), and Hotteling's multivariate two sample T2-Test was significant (P < 0.01). Patients were then sampled according to the PSA/FT ratio in group 1 (PSA/FT <= 0.20), group 2 (PSA/FT > 0.20 and <= 0.40), and group 3 (PSA/FT > 0.40), and Box's M-Test comparing the covariance matrices of the 3 groups differed significantly (P < 0.001). Finally, patients were sampled according to the PSA/FT ratio in 6 groups, and Box's M-Test was again significant (P < 0.001). CONCLUSIONS: The PSA to FT ratio is the growing rate parameter expressing different biology patterns and assessing different groups of prostate cancer patients. In our opinion, the results of the present study might have wide applications in understanding, assessing and planning prostate cancer studies including basic science, screening, assessing risk of the disease, predicting disease stage as well natural history after a planned treatment involving biochemical recurrence, progression, hormone refractory prostate cancer and disease-specific survival. PMID- 19902379 TI - Single-knot running suture anastomosis (one-knot pyeloplasty) for laparoscopic dismembered pyeloplasty: training model on a porcine bladder and clinical results. AB - OBJECTIVES: The one-knot running ureteropelvic anastomosis is a modification of the single-knot running suture that was previously described for urethrovesical anastomosis. In this study, we present a novel porcine model for laparoscopic pyeloplasty training and report the results obtained in patients who underwent one-knot pyeloplasty. MATERIALS AND METHODS: A porcine bladder was used for the ureteropelvic junction simulation in this training model. The laparoscopic one knot pyeloplasty technique was attempted by five laparoscopic surgeons using this model, and the technique was then incorporated into clinical practice. The data of all patients who underwent this procedure between January 2006 and February 2008 were evaluated. RESULTS: The one-knot pyeloplasty technique was easily applied in a short time by laparoscopic surgeons in a novel porcine pyeloplasty model. The participants successfully performed a watertight anastomosis in the porcine bladder model, completing the task in <30 min by the fifth attempt. The time required to succeed before and after training decreased by 20.8% (P = 0.01). In the clinical portion, 40 laparoscopic pyeloplasty procedures were performed with this technique, and the mean time to complete the anastomosis was 27.1 min (range: 12-41). This time was concordant with the final anastomosis time performed in the pelvitrainer. CONCLUSIONS: The one-knot pyeloplasty is feasible and reproducible, and it overcomes the obstacles caused by multiple intracorporeal knots when performing laparoscopic pyeloplasty. Furthermore, the porcine bladder model that we introduce herein is a readily available and simple model to refine suturing techniques for pyeloplasty. PMID- 19902381 TI - The anti-necrosis role of hypoxic preconditioning after acute anoxia is mediated by aldose reductase and sorbitol pathway in PC12 cells. AB - It has been demonstrated that hypoxic preconditioning (HP) enhances the survival ability of the organism against the subsequent acute anoxia (AA). However, it is not yet clear whether necrosis induced by AA can be prevented by HP, and what are the underlying mechanisms. In this study, we examined the effect of HP (10% O(2), 48 h) on necrosis induced by AA (0% O(2), 24 h) in PC12 cells. We found that HP delayed the regulatory volume decrease and reduced cell swelling after 24 h of exposure to AA. Since aldose reductase (AR) is involved in cell volume regulation, we detected AR mRNA expression with reverse transcription-polymerase chain reaction (RT-PCR) techniques. The AR mRNA level was dramatically elevated by HP. Furthermore, an HP-induced decrease in cell injury was reversed by berberine chloride (BB), the inhibitor of AR. In addition, sorbitol synthesized from glucose catalyzed by AR is directly related to cell volume regulation. Subsequently, we tested sorbitol content in the cytoplasm. HP clearly elevated sorbitol content, while BB inhibited the elevation induced by HP. Further study showed that a strong inhibitor of sorbitol permease, quinidine, completely reversed the protection induced by HP after AA. These data provide evidence that HP prevents necrosis induced by AA and is mediated by AR and sorbitol pathway. PMID- 19902380 TI - The chloroplastic thiol reducing systems: dual functions in the regulation of carbohydrate metabolism and regeneration of antioxidant enzymes, emphasis on the poplar redoxin equipment. AB - The post-translational modification consisting in the formation/reduction of disulfide bonds has been the subject of intense research in plants since the discovery in the 1970s that many chloroplastic enzymes are regulated by light through dithiol-disulfide exchange reactions catalyzed by oxidoreductases called thioredoxins (Trxs). Further biochemical and proteomic studies have considerably increased the number of target enzymes and processes regulated by these mechanisms in many sub-cellular compartments. Recently, glutathionylation, a modification consisting in the reversible formation of a glutathione adduct on cysteine residues, was proposed as an alternative redox regulation mechanism. Glutaredoxins (Grxs), proteins related to Trxs, are efficient catalysts for deglutathionylation, the opposite reaction. Hence, the Trxs- and Grxs-dependent pathways might constitute complementary and not only redundant regulatory processes. This article focuses on these two multigenic families and associated protein partners in poplar and on their involvement in the regulation of some major chloroplastic processes such as stress response, carbohydrate and heme/chlorophyll metabolism. PMID- 19902382 TI - Synthesis of cholesterol-conjugated magnetic nanoparticles for purification of human paraoxonase 1. AB - Human serum paraoxonase 1 (PON1) is known as an antioxidant and is also involved in the detoxification of many compounds. In this study, a novel purification strategy was employed to purify the PON1 by using cholesterol-conjugated magnetic nanoparticles. Magnetic nanoparticles were synthesized and conjugated with cholesterol through diazotized p-aminohippuric acid. In Fourier transform infrared spectrum of cholesterol-p-aminohippuric acid-Fe(3)O(4) nanoparticles, the appearance of peaks at 3,358.3, 1,645 cm(-1), and at 2,334.9 cm(-1) confirmed the conjugation. The molecular weight of purified PON1 was nearly 45 kDa on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE), and isoelectric point was 5.3. The specific activity was 438 U mg(-1) protein, and the purification fold was 515 with 73% yield. The K (m) values were 1.3 and 0.74 mM with paraoxon and phenyl acetate, respectively. Western blot of 2D-PAGE confirmed the homogeneity and stability of the enzyme. Mg(+2), Mn(+2), glycerol, (NH(4))(2)SO(4), PEG 6000, Triton X-100, and phenylmethylsulfonyl fluoride did not show any effect on activity. Pb(+2), Co(+2), Zn(2+), ethanol, beta mercaptoethanol, and acetone reduced the activity while Ni(2+), Cd(2+), Cu(2+), iodoacetic acid, SDS, dimethylformamide, DMSO inhibited the activity. In vitro enzyme activity was slightly reduced by acetyl salicylic and acetaminophen and reduced 50% with amino glycosides and ampicillin antibiotics at concentrations of 0.6 and 30 mg ml(-1), respectively. This is the first report for the synthesis of cholesterol-conjugated magnetic nanoparticles for simple purification of PON1 enzyme. PMID- 19902383 TI - Purification, molecular cloning, and biochemical characterization of subtilisin JB1 from a newly isolated Bacillus subtilis JB1. AB - An extracellular gelatinolytic enzyme obtained from the newly isolated Bacillus subtilis JB1, a thermophilic microorganism relevant to the aerobic biodegradation process of fish-meal production, was purified via ammonium sulfate precipitation, Sephadex G-200 Gel filtration chromatography, and one-dimensional gel electrophoresis separation and subsequently identified via peptide mass fingerprinting and chemically assisted fragmentation matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The subtilisin JB1 gene was sequenced and its recombinant protein prosubtilisin JB1 was expressed in Escherichia coli, and the purified prosubtilisin JB1 (62 kDa) protein was digested with gelatin, bovine serum albumin, azocasein, fibrinogen, and the fluorogenic peptide substrate Ala-Ala-Phe-7-amido-4-methylcoumarin hydrochloride, whereas the serine protease inhibitors phenylmethylsulfonyl fluoride and chymostatin completely inhibited its enzyme activity at an optimal pH of 7.5. Thus, our results show that subtilisin JB1 may serve as a potential source material for use in industrial applications of proteolytic enzymes and microorganisms for fishery waste degradation and fish by-product processing. PMID- 19902385 TI - Intracerebroventricular transplantation of human mesenchymal stem cells induced to secrete neurotrophic factors attenuates clinical symptoms in a mouse model of multiple sclerosis. AB - Stem cell-based therapy holds great potential for future treatment of multiple sclerosis (MS). Bone marrow mesenchymal stem cells (MSCs) were previously reported to ameliorate symptoms in mouse MS models (experimental autoimmune encephalomyelitis, EAE). In this study, we induced MSCs to differentiate in vitro into neurotrophic factor-producing cells (NTFCs). Our main goal was to examine the clinical use of NTFCs on EAE symptoms. The NTFCs and MSCs were transplanted intracerebroventricularly (ICV) to EAE mice. We found that NTFCs transplantations resulted in a delay of symptom onset and increased animal survival. Transplantation of MSCs also exerted a positive effect but to a lesser extent. In vitro analysis demonstrated the NTFCs' capacity to suppress mice immune cells and protect neuronal cells from oxidative insult. Our results indicate that NTFCs transplanted ICV delay disease symptoms of EAE mice, possibly via neuroprotection and immunomodulation, and may serve as a possible treatment to MS. PMID- 19902386 TI - A giant bilateral calcified chronic subdural hematoma. AB - BACKGROUND: A calcified chronic subdural hematoma is a rare disease and its neuroradiological presentation is variable. The degree of calcification extends from thin calcified inner membranes to dense calcification and even ossification of the hematoma. Previous reports described a maximum of two hematoma cavities with calcified inner hematoma membranes. METHODS: Neuroimaging report with illustrative computerized tomography images. RESULTS: A patient with a bilateral symptomatic calcified chronic subdural hematoma, or so-called "armoured brain", was admitted to our intensive care unit with clinical signs of increased intracranial pressure. Computerized cranial tomography demonstrated multiple bilaterally located hematoma cavities with thin calcified inner membranes. After neurosurgical intervention by bilateral burr hole trepanation, clinical symptoms improved. CONCLUSIONS: Our case of a calcified chronic subdural hematoma presents with an uncommon imaging pattern with more than four hematoma cavities bounded by predominantly convex- and concave-configured thin calcified inner membranes. PMID- 19902388 TI - A modified protocol for RNA isolation from high polysaccharide containing Cupressus arizonica pollen. Applications for RT-PCR and phage display library construction. AB - RNA isolation is the first step in the study of gene expression and recombinant protein production. However, the isolation of high quantity and high-quality RNA from tissues containing large amounts of polysaccharides has proven to be a difficult process. Cupressus arizonica pollen, in addition to containing high polysaccharide levels, is a challenging starting material for RNA isolation due to the roughness of the pollen grain's walls. Here, we describe an improved technique for RNA isolation from C. arizonica pollen grains. The protocol includes a special disruption and homogenization process as well as a two-step modified RNA isolation technique which consists of an acid phenol extraction followed by a final cleanup using a commercial kit. Resulting RNA proved to be free of contaminants as determined by UV spectrophotometry. The quality of the RNA was analyzed on a bioanalyzer and showed visible 25S and 18S bands. This RNA was successfully used in downstream applications such as RT-PCR and phage display library construction. PMID- 19902387 TI - Manganese flux across the blood-brain barrier. AB - Manganese (Mn) is essential for brain growth and metabolism, but in excess can be a neurotoxicant. The chemical form (species) of Mn influences its kinetics and toxicity. Significant Mn species entering the brain are the Mn(2+) ion and Mn citrate which, along with Mn transferrin, enter the brain by carrier-mediated processes. Although the divalent metal transporter (DMT-1) was suggested to be a candidate for brain Mn uptake, brain Mn influx was not different in Belgrade rats, which do not express functional DMT-1, compared to controls. Brain Mn influx was not sodium dependent or dependent on ATP hydrolysis, but was reduced by mitochondrial energy inhibitors. Mn and Fe do not appear to compete for brain uptake. Brain Mn uptake appears to be mediated by a Ca uptake mechanism, thought to not be a p-type ATPase, but a store-operated calcium channel. Efflux of Mn from the brain was found to be slower than markers used as membrane impermeable reference compounds, suggesting diffusion mediates brain Mn efflux. Owing to carrier-mediated brain Mn influx and diffusion-mediated efflux, slow brain Mn clearance and brain Mn accumulation with repeated excess exposure would be predicted, and have been reported. This may render the brain susceptible to Mn induced neurotoxicity from excessive Mn exposure. PMID- 19902391 TI - [Role of the cannabinoids in glaucoma]. PMID- 19902390 TI - Pharmacogenetics and pharmacoepigenetics of gemcitabine. AB - Gemcitabine (2',2'-difluoro 2'deoxycytidine, dFdC) is an analog of cytosine with distinctive pharmacological properties and a wide antitumor-activity spectrum. The pharmacological characteristics of gemcitabine are unique because two main classes of genes are essential for its antitumor effects: membrane transporter protein-coding genes, whose products are responsible for drug intracellular uptake, as well as enzyme-coding genes, which catalyze its activation and inactivation. The study of the pharmacogenetics and pharmacoepigenetics of these two gene classes is greatly required to optimize the drug's therapeutic use in cancer. This review aims to provide an update of genetic and epigenetic bases that may account for interindividual variation in therapeutic outcome exhibited by gemcitabine. PMID- 19902392 TI - [Intraocular lens power calculation in complicated cases]. PMID- 19902393 TI - [New developments in glaucoma medical treatment]. AB - The medical treatment of glaucoma has undergone significant development in recent years. Research in this field is focused on improving pre-existing drugs and on the development of new molecules. In relation to commercial drugs, there is a trend to improve local tolerance, using less toxic preservatives as in the case of sofZIA in travoprost, and eliminating the preservatives as in tafluprost. The development of new, fixed combinations of commercial drugs could also enhance their administration and therapeutic compliance. There is also intense research activity in the search for new therapeutic groups for glaucoma treatment. Calcium channel-blockers such as lomerizine do not seem to affect systemic hypotension, while topical calcium-blockers like flunarizine and iganidipine are also under research. Endothelin 1 antagonists such as sulfisoxazole and bunazosine could be also useful in the treatment of glaucoma. In the renin angiotensin system, angiotensin (1-7) and olmesartan are under investigation for use in glaucoma patients. Trabecular drugs such as Rho-kinase inhibitors could be effective on the pathogenic mechanism of primary open angle glaucoma. Finally, topical mifepristone, an antagonist of glucocorticoid receptors, is under evaluation for corticosteroid-induced elevated intraocular pressure (Arch Soc Esp Oftalmol 2009; 84: 491-500). PMID- 19902394 TI - [Outcomes of inferior access external dacryocystorhinostomy performed by residents]. AB - PURPOSE: To assess the outcomes of external dacryocystorhinostomy (DCREx) performed by residents at the Hospital Universitari de Bellvitge, comparing the cases operated on by residents and the cases operated on by staff surgeons. METHODS: We performed a retrospective study between January 2006 and December 2007 of all DCREx performed at our hospital. The operations involved 65 cases from 54 patients: 16 operations were performed by residents and 49 by staff surgeons. The cases were divided into two groups: cases operated on by residents and cases operated on by staff surgeons, and the following variables were studied: epidemiologic characteristics of the patients (age, sex), side of intervention, antecedents of lacrimal sac inflammation, type of anesthesia, intrasurgical and postsurgical complications and results. <> was defined as the possibility to pass physiologic serum through the lacrimal pathway and <> if the patient in addition was without symptoms of epiphora. RESULTS: The procedure was successful in 88% of cases: 80% were classified as <> while only 8% were considered <>. The operations performed by residents were successful in 81% of cases (75% <> and 6% <>), whereas staff surgeons achieved success in 90% of cases (82% <> - 8% <>). CONCLUSION: The results show that residents with suitable training and supervision can achieve good success rates approaching those of staff surgeon outcomes, without additional complications (Arch Soc Esp Oftalmol 2009; 84: 501-506). PMID- 19902395 TI - [Ranibizumab as treatment for myopic choroidal neovascularization]. AB - OBJECTIVE: To evaluate the efficacy of intravitreal Ranibizumab as treatment for choroidal neovascularisation due to pathological myopia. MATERIALS AND METHODS: A retrospective, non-comparative study of 18 eyes treated with intravitreal injections of Ranibizumab. Ten eyes had been treated previously with photodynamic therapy and eight received Ranibizumab as first therapy. After thorough ophthalmologic examination, fluorescein angiography (FAG) and optical coherence tomography (OCT), intraocular injection of Ranibizumab was performed. In subsequent monthly follow ups and taking into account visual acuity, presence or absence of metamorphopsia, biomicroscopy and OCT examination, further treatment was decided. RESULTS: Eighteen eyes from 16 patients were finally included. Patients were followed up for a minimum of 6 months. The mean age at initial treatment was 56.4 years. Mean refractive error was -13.3 diopters. Regarding FAG, all neovascular membranes were classical and sub or juxtafoveal localised. At the end of the sixth month after treatment fourteen eyes (77.7%) showed better visual acuity ranging from one or more lines on the Snellen chart, eleven eyes (61.1%) improved two lines or more, three eyes (16.6%) did not show any change and one eye (5.5%) worsened by one line. At 6 months the mean best-corrected visual acuity improved from 0.25 to 0.46 (p= 0.001). The mean central macular thickness decreased from 344.9 to 212.6 (p=0.015). CONCLUSIONS: Ranibizumab may be a good therapeutic option as treatment for choroidal neovascularisation due to pathological myopia; it improved visual acuity and anatomical features, even in non-responders to photodynamic therapy (Arch Soc Esp Oftalmol 2009; 84: 507-514). PMID- 19902396 TI - [Treatment of lacrimal duct obstruction with a Tear-Leader stent]. AB - PURPOSE: To compare the effectiveness and patency of the TearLeader stent (PBN) for treatment of lacrimal system obstruction. METHODS: A prospective study of 68 patients referred from the ophthalmology department with suspicion of lacrimal system obstruction. We placed 74 TearLeader PBN stents. Inclusion criteria were: complete obstruction of lacrimal duct with canalicula and lacrimal puncta patency, and absence of acute infection. We studied the clinical improvement by means of an opinion survey, and the patency of stents was evaluated with Kaplan Meier survival curves. RESULTS: Follow-up of patients was two years. A painful procedure was reported in 10% of cases. Minor dacryocystitis appeared in 18.9% of cases, while complete resolution of epiphora was confirmed in 77% of cases (23% of cases showed grade I epiphora). Patency of stents: median patency 490 days (15 months), range 11 to 730 days; 1 year after stent placement patency was 0.51 and long term patency rate for 2 years was 0.31. Opinion survey of the 68 patients: satisfaction with the technique, the procedure and prosthesis placement was 41%; satisfaction whilst the stent remained patent was 60.8%. CONCLUSION: Tearleader stent placement is easy to perform and comfortable for patients. TearLeader placement gave a patency outcome similar to other stents (Arch Soc Esp Oftalmol 2009; 84: 515-522). PMID- 19902397 TI - [Complications of uveitis: prevalence and risk factors in a series of 398 cases]. AB - OBJECTIVE: To estimate the rate of complications and establish risk factors for their development in patients with uveitis. METHODS: Prospective study of a cohort of 398 patients (413 eyes) that were reviewed from January 2000 to October 2007 and monitored during a period of at least one month. Demographic data, laterality, course (acute, subacute, chronic), location, diagnosis (idiopathic, infectious, ocular origin, systemic origin, others) and development of complications (cataract, cystoid macular edema, macular complications, retinal complications and others) were evaluated. RESULTS: After a mean follow-up of 35.2 months (range 1-160), 25% of patients developed one or more of the following complications: cataract 8.5%, macular edema 6.1%, macular complications (epiretinal membranes, choroidal neovascular membranes, macular necrosis) 4.1%, retinal complications (retinal detachment, retinal neovascularization, vitreous hemorrhage, retinal tears, retinal vascular occlusions) 4.1%, glaucoma or ocular hypertension 3.6%, and others 1.5%. Risk factors for development of uveitis complications were chronic course OR 6.37 (3.25-12.47) P <0.0001, bilaterality OR 1.98 (1.03-3.08) P =0.039, subacute course OR 1.94 (0.02-4.08) P=0.08, and panuveitis OR 1,92 (0.99-4.80) P=0.161. CONCLUSIONS: In our patients, cases involving chronic and bilateral uveitis were more prone to develop complications, with cataracts being the most frequent complication (Arch Soc Esp Oftalmol 2009; 84: 523-528). PMID- 19902398 TI - [Fluorescein angiography with Retcam in incontinentia pigmenti: a case report]. AB - CASE REPORT: The ophthalmic examination and results of fluorescein angiography using Retcam II are described in a patient with Incontinentia Pigmenti (IP). DISCUSSION: Angiography fluorescein is extremely valuable in detecting vascular lesions that were invisible with ordinary ophthalmoscopy. Retcam II allows documentation of these lesions which is very useful for diagnosis, treatment and follow-up of this disease (Arch Soc Esp Oftalmol 2009; 84: 529-532). PMID- 19902399 TI - [Management of Descemet's membrane rupture by intracameral injection of SF6 in acute hydrops]. AB - CASE REPORT: Hydrops occurring in the eye secondary to keratoconus, grade 4. It was managed by sulfur hexafluoride (SF6) gas injected into the anterior chamber, with an early resolution of corneal edema obtained. DISCUSSION: Intervention with intracameral SF6 injection has proven to be a safe and effective therapy for early reduction of corneal edema in an eye with Descemet's membrane detachment and acute hydrops (Arch Soc Esp Oftalmol 2009; 84: 533-536). PMID- 19902402 TI - Expression of the lipid transporters ABCA3 and ABCA1 is diminished in human breast cancer tissue. AB - ATP-binding cassette transporters ABCA3 and ABCA1 are related to a differentiated, lipid-secreting phenotype of type II pneumocytes. Since mammary gland epithelial cells also show pronounced lipid metabolism and secretion, we investigated the expression of these proteins in normal as well as in neoplastic breast tissue. Normal human breast tissue, breast cancer cell lines, and 162 tumor samples of patients with primary unilateral invasive breast cancer were analyzed for ABCA3 and ABCA1 protein expression by immunohistochemistry using tissue microarrays. Strong ABCA3 and ABCA1 expression was found in the inner layer of normal mammary gland epithelium. Concurrent cytoplasmic ABCA3 and ABCA1 immunoreactivity was found in 9 of 11 breast cancer cell lines. ABCA3 and ABCA1 were shown to be differentially expressed in human breast cancer. Loss of ABCA3 staining was significantly associated with positive nodal status and negative progesterone receptor expression. In multivariate analysis, diminished ABCA3 expression proved to be a significant, independent and adverse risk factor for tumor recurrence. ABCA1 expression was associated with positive lymph nodes, but not significantly associated with tumor recurrence or breast cancer-specific survival. ABCA3 and ABCA1 are strongly expressed in normal mammary gland epithelium. Decreased ABCA3 expression in breast cancer seems to be associated with poor prognosis. PMID- 19902403 TI - Early infant diet in children at high risk for type 1 diabetes. AB - Infant diet affects health and development. The aim of our study was to investigate WHO infant feeding compliance in children who have a first degree family history of type 1 diabetes (T1D). One hundred and fifty children who were first degree relatives of patients with T1D were intensively followed from birth in the BABYDIET intervention study. Infant feeding, infections, and medication were recorded daily by participating families. Weight and length of children were obtained from paediatric records. Only 5% of the families followed the WHO recommendations for infant feeding that include full breastfeeding for at least 6 months (18.8% of children) and introduction of complementary foods under continued breastfeeding thereafter (22.2% of children). Maternal age in the first quartile (<29 years; p<0.0001), and maternal smoking (p<0.0001) were associated with an earlier introduction of solid food and reduced breastfeeding. Full breastfeeding > or =6 months was associated with reduced frequency of gastrointestinal infections (12 vs. 38%, p=0.02) and antibiotic treatment (24 vs. 48%, p=0.04). Our findings indicate that WHO infant feeding recommendations were poorly followed by families with a family history of T1D. Action to improve levels of infant feeding behaviour is essential, especially among young mothers with T1D. PMID- 19902404 TI - Complete absence of the deep inferior epigastric artery: an increasingly detected anomaly detected with the use of advanced imaging technologies. PMID- 19902405 TI - Double arterialized free jejunal flap. AB - In a standard free jejunal transfer, one artery and one vein are anastomosed. However, when raising the jejunal flap, a one-segment jejunum sometimes has two arteries and one accompanying vein as a vascular pedicle. Free jejunal transfer in which two arteries and one vein are anastomosed was designed. We report on the safety and advantages of using this artery-dominant transfer when performing microvascular anastomosis. This technique was used when a one-segment jejunum had two arteries and an accompanying vein. Eight patients underwent this arterial supercharged free jejunal transfer. All flaps survived, and no complications developed except for two cases of intraoperative thrombosis before the procedure. It is important to transfer the artery-rich graft into the same physiological environment by reconstructing the similar hemodynamics. The grafts can be transferred without harm. This artery-dominant method can be an option when conditions are unfavorable for safer jejunal transfer. PMID- 19902406 TI - Intraoperative hemodynamic evaluation of the radial and ulnar arteries during free radial forearm flap procedure. AB - The purpose of this prospective study was to assess the blood flow of the radial and ulnar arteries before and after radial forearm flap raising. Twenty-two patients underwent radial forearm microvascular reconstruction for leg soft tissue defects. Blood flow of the radial, ulnar, and recipient arteries was measured intraoperatively by transit-time and ultrasonic flowmeter. In the in situ radial artery, the mean blood flow was 60.5 +/- 47.7 mL/min before, 6.7 +/- 4.1 mL/min after raising the flap, and 5.8 +/- 2.0 mL/min after end-to-end anastomosis to the recipient artery. In the ulnar artery, the mean blood flow was 60.5 +/- 43.3 mL/min before harvesting the radial forearm flap and significantly increased to 85.7 +/- 57.9 mL/min after radial artery sacrifice. A significant difference was also found between this value and the value of blood flow in the ulnar and radial arteries pooled together ( P < 0.05). The vascular resistance in the ulnar artery decreased significantly after the radial artery flap raising (from 2.7 +/- 3.1 to 1.9 +/- 2.2 peripheral resistance units, P = 0.010). The forearm has a conspicuous arterial vascularization not only through the radial and ulnar arteries but also through the interosseous system. The raising of the radial forearm flap increases blood flow and decreases vascular resistance in the ulnar artery. PMID- 19902407 TI - Treatment of ischial pressure sores using a modified gracilis myofasciocutaneous flap. AB - Despite the availability of a variety of flap reconstruction options, ischial pressure sores continue to be the most difficult pressure sores to treat. This article describes a successful surgical procedure for the coverage of ischial ulcers using a modified gracilis myofasciocutaneous flap. From August 2000 to April 2004, 12 patients with ischial sores were enrolled in the study. All patients underwent early aggressive surgical debridement followed by surgical reconstruction with a modified gracilis myofasciocutaneous flap. The follow-up period ranged from 13 to 86 months, with a mean of 44 months. Overall, 91.7% of the flaps (11 of 12) survived primarily. Partial flap necrosis occurred in one patient. Primary wound healing occurred without complications at both the donor and recipient sites in all cases. In one patient, grade II ischial pressure sores recurred 13 months after the operation. There was no recurrence in other 11 patients. A modified gracilis myofasciocutaneous flap provides a good cover for ischial pressure sores. Because it is easy to use and has favorable results, it can be used in the primary treatment for large and deep ischial pressure sores. PMID- 19902408 TI - Reconstruction of extensive composite oromandibular defects with simultaneous free anterolateral thigh fasciocutaneous and fibular osteocutaneous flaps. AB - Massive composite defects of the face are difficult to reconstruct. Such defects are usually created after ablation of advanced cancers of the head and neck region. The use of a free fibular osteocutaneous flap for the bone and mucosal lining of the oral cavity and anterolateral thigh flap for the outer cutaneous lining are well established. We present our experience of using these two flaps simultaneously in the reconstruction of such defects and to evaluate the outcome. There were a total of 10 patients in our study. Their average age was 48.8 years. All had squamous cell carcinoma of the oral cavity. Their pathological stages were mostly stage T4 with only one case being T3. Flap survival was 100%. The application of dual free flaps, though technically more demanding, allows good orientation of the flaps. Seven patients maintained a good functional outcome. They were able to eat a soft diet. Their speech was easily comprehensible. The combination of a free anterolateral thigh flap with vascularized fibular osteocutaneous flap can be performed safely with adequate functional outcome. This combination of flaps should be considered for this group of patients. PMID- 19902409 TI - Comprehensive approach in surgical reconstruction of facial nerve paralysis: a 10 year perspective. AB - Facial paralysis presents diverse functional and aesthetic abnormalities. Reconstruction may be achieved by several methods. We reviewed the management and outcome of facial paralysis patients to establish principles on which a comprehensive reconstructive approach may be based. Records were reviewed of all patients operated for facial paralysis at our institution between 1998 and 2007. Ninety-five patients were included, of which 15 patients had static reconstruction alone, and 80 patients had dynamic reconstruction. Presented is our experience in reconstruction of facial paralysis over the past decade, delineating a comprehensive approach to this condition. Various surgical techniques are described. PMID- 19902410 TI - Analysis of a series of isomeric oligosaccharides by energy-resolved mass spectrometry: a challenge on homobranched trisaccharides. AB - Glycans exist as part of glycoproteins and glycolipids, which are involved in a variety of biological functions. The analysis of glycan structures, particularly that of structural isomers, is fundamentally important since isomeric glycans often show distinct functions; however, a method for their structural elucidation has not yet been established. Anomeric configurations, linkage positions and branching are the major factors in glycans and their alteration results in a large diversity of glycan structures. The analysis of vicinally substituted oligosaccharides is extremely difficult because the product ions formed in tandem mass spectrometry (MS/MS) often have the same m/z values. In our endeavor to address the issue, we analyzed a series of homo-substituted trisaccharides consisting only of glucose by collision-induced dissociation (CID), especially energy-resolved mass spectrometry (ERMS). It was found that these structurally related glycans could be distinguished by taking advantage of differences in their activation energies in ERMS. PMID- 19902411 TI - Changes in the natural abundance of 13CO2/12CO2 in breath due to lipopolysacchride-induced acute phase response. AB - The natural abundance of carbon-13 in blood proteins increases during the cachectic state and may be a biomarker for disease status. We hypothesized a corresponding drop in the relative abundance of 13C in breath CO2. Using the lipopolysacchride (LPS)-induced endotoxemia model of the acute cachectic state, we demonstrated that the acute phase response causes shifts in the stable isotopes of carbon in exhaled CO2 (13CO2/12CO2 delta value) shortly after administration of LPS while glucocorticoid treatment does not. Mice were injected with LPS and stable isotopes of blood amino acids and carbon in exhaled CO2 were monitored. An increase in the relative isotopic mass of serum alanine, proline and threonine was observed at 3 h after LPS injection. Breath delta values began dropping immediately after administration of LPS, and were 4-5 delta values lower than those of the control animals by 2.5 h after injection. A corresponding drop in delta value was not observed with dexamethasone treatment. Thus protein synthesis during the acute phase response probably caused the fractionation of stable isotopes observed in the plasma amino acids and in exhaled breath 13CO2 delta values. The exhaled breath 13CO2 delta value may be a valuable real-time biomarker of cachexia associated with an acute phase response due to endotoxemia. PMID- 19902412 TI - Metabolomics profiling of extracellular metabolites in recombinant Chinese Hamster Ovary fed-batch culture. AB - A metabolomics-based approach was used to time profile extracellular metabolites in duplicate fed-batch bioreactor cultures of recombinant Chinese Hamster Ovary (CHO) cells producing monoclonal IgG antibody. Culture medium was collected and analysed using a high-performance liquid chromatography (HPLC) system in tandem with an LTQ-Orbitrap mass spectrometer. An in-house software was developed to pre process the LC/MS data in terms of filtering and peak detection. This was followed by principal component analysis (PCA) to assess variance amongst the samples, and hierarchical clustering to categorize mass peaks by their time profiles. Finally, LC/MS2 experiments using the LTQ-Orbitrap (where standard was available) and SYNAPT HDMS (where standard was unavailable) were performed to confirm the identities of the metabolites. Two groups of identified metabolites were of particular interest; the first consisted of metabolites that began to accumulate when the culture entered stationary phase. The majority of them were amino acid derivatives and they were likely to be derived from the amino acids in the feed media. Examples included acetylphenylalanine and dimethylarginine which are known to be detrimental to cell growth. The second group of metabolites showed a downward trend as the culture progressed. Two of them were medium components--tryptophan and choline, and these became depleted midway into the culture despite the addition of feed media. The findings demonstrated the potential of utilizing metabolomics to guide medium design for fed-batch culture to potentially improve cell growth and product titer. PMID- 19902413 TI - Analysis of [U-13C6]glucose in human plasma using liquid chromatography/isotope ratio mass spectrometry compared with two other mass spectrometry techniques. AB - The use of stable isotope labelled glucose provides insight into glucose metabolism. The 13C-isotopic enrichment of glucose is usually measured by gas chromatography/mass spectrometry (GC/MS) or gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). However, in both techniques the samples must be derivatized prior to analysis, which makes sample preparation more labour intensive and increases the uncertainty of the measured isotopic composition. A novel method for the determination of isotopic enrichment of glucose in human plasma using liquid chromatography/isotope ratio mass spectrometry (LC/IRMS) has been developed. Using this technique, for which hardly any sample preparation is needed, we showed that both the enrichment and the concentration could be measured with very high precision using only 20 microL of plasma. In addition, a comparison with GC/MS and GC/IRMS showed that the best performance was achieved with the LC/IRMS method making it the method of choice for the measurement of 13C isotopic enrichment in plasma samples. PMID- 19902414 TI - Mass spectrometry and UV-VIS spectrophotometry of ruthenium(II) [RuClCp(mPTA)2](OSO2CF3)2 complex in solution. AB - Ruthenium(II) complexes are of great interest as a new class of cancerostatics with advantages over classical platinum compounds including lower toxicity. The stability of the [RuClCp(mPTA)2](OSO2CF3)2 complex (I) (Cp cyclopentadienyl, mPTA N-methyl 1,3,5-triaza-7-phosphaadamantane) in aqueous solution was studied using spectrophotometry, matrix-assisted laser desorption/ionization (MALDI) and laser desorption/ionization (LDI) time-of-flight (TOF) mass spectrometry (MS). Spectrophotometry proves that at least three different reactions take place in water. Dissolution of I leads to fast coordination of water molecules to the Ru(II) cation and then slow hydrolysis and ligand exchange of chloride and mPTA with water, hydroxide or with trifluoromethane sulfonate itself. Via MALDI and LDI of the hydrolyzed solutions the formation of singly positively charged ions of general formula RuCl(p)(Cp)(q)(mPTA)(r)(H2O)(s)(OH)(t) (p = 0-1, q = 0-1, r = 0-2, s = 0-5, t = 0-2) and of some fragment ions was shown. The stoichiometry was determined by analyzing the isotopic envelopes and computer modelling. The [RuClCp(mPTA)2](OSO2CF3)2 complex can be stabilized in dilute hydrochloric acid or in neutral 0.15 M isotonic sodium chloride solution. PMID- 19902415 TI - Online nanoelectrospray/high-field asymmetric waveform ion mobility spectrometry as a potential tool for discovery pharmaceutical bioanalysis. AB - Nanoelectrospray ionization (nESI) coupled online with high-field asymmetric waveform ion mobility spectrometry (FAIMS) for small molecule analysis in a discovery pharmaceutical setting was examined. A conventional capillary pump, autosampler and nESI source were used to introduce samples directly into the FAIMS device. The FAIMS device was used to separate gas-phase ions on a timescale that was compatible with the mass spectrometer. The capability of the nESI-FAIMS combination to efficiently remove metabolite interferences from the parent drug, and reduce ion suppression effects, was demonstrated. On average, 85% of the signal intensity obtained from a neat sample was preserved in the extracted plasma samples. Standard curves were prepared for several compounds. Linearity was obtained over approximately 3 to 4 orders of magnitude. Comparison of results from nESI-FAIMS with those from conventional LC/MS for a mouse pharmacokinetic study yielded concentration values differing by no more than 30%. PMID- 19902416 TI - Analysis of low-concentration gas samples with continuous-flow isotope ratio mass spectrometry: eliminating sources of contamination to achieve high precision. AB - Developments in continuous-flow isotope ratio mass spectrometry have made possible the rapid analysis of delta13C in CO2 of small-volume gas samples with precisions of < or = 0.1 per thousand. Prior research has validated the integrity of septum-capped vials for collection and short-term storage of gas samples. However, there has been little investigation into the sources of contamination during the preparation and analysis of low-concentration gas samples. In this study we determined (1) sources of contamination on a Gasbench II, (2) developed an analytical procedure to reduce contamination, and (3) identified an efficient, precise method for introducing sample gas into vials. We investigated three vial filling procedures: (1) automated flush-fill (AFF), (2) vacuum back-fill (VBF), and (3) hand-fill (HF). Treatments were evaluated based on the time required for preparation, observed contamination, and multi-vial precision. The worst-case observed contamination was 4.5% of sample volume. Our empirical estimate showed that this level of contamination results in an error of 1.7 per thousand for samples with near-ambient CO2 concentrations and isotopic values that followed a high-concentration carbonate reference with an isotope ratio of -47 per thousand (IAEA-CO-9). This carry-over contamination on the Gasbench can be reduced by placing a helium-filled vial between the standard and the succeeding sample or by ignoring the first two of five sample peaks generated by each analysis. High precision (SD < or = 0.1 per thousand) results with no detectable room-air contamination were observed for AFF and VBF treatments. In contrast, the precision of HF treatments was lower (SD > or = 0.2 per thousand). VBF was optimal for the preparation of gas samples, as it yielded faster throughput at similar precision to AFF. PMID- 19902417 TI - Kinetics of albumin homocysteinylation measured with matrix-assisted laser/desorption ionization mass spectrometry versus with a radioactive tracer. AB - Homocysteinylation is a post-translational protein modification which involves homocysteine-thiolactone and may be responsible for many pathophysiological changes secondary to hyperhomocysteinemia. Therefore, methods to measure protein homocysteinylation in intact biological samples are required. We tested whether matrix assisted-laser/desorption ionization mass spectrometry (MALDI-MS) can detect time- and dose-dependent changes in in vitro homocysteine-thiolactone binding to human serum albumin. We have compared this method with a 35S thiolactone radioactive binding assay. Incubations with and without dithiothreitol allowed measurement of the amide-linked and disulfide-linked thiolactone-protein adducts, respectively. A good correspondence in time- and dose-dependent protein-thiolactone formation was observed between the two methods. A maximum of 9 to 12 thiolactone residues were bound to each albumin molecule. The 35S-thiolactone bound albumin tightly, particularly at the lowest concentrations, with approximately 70% of the binding amide-linked. Although the results of the two methods were rather similar, the radioactive method appears to be more sensitive than the MALDI-MS technique. PMID- 19902418 TI - Investigation of the identification point system adaptation in cocaine, benzoylecgonine and ecgonine methyl ester using a single quadrupole mass spectrometer. AB - At present, no official criteria exist for drug identification using single quadrupole mass spectrometers although the European Union (EU) criteria for compound identification have been adopted. These criteria are evaluated with respect to the confirmation of cocaine and its metabolites by single quadrupole liquid chromatography/mass spectrometry (LC/MS) and problems are highlighted. Spiked samples, proficiency testing samples, certified reference materials and samples from real cases that had screened positive for cocaine derivatives by immunoassay were subjected to confirmation by LC/MS using single ion monitoring with in-source fragmentation. The EU criteria for compound identification were applied for the confirmation of cocaine, benzoylecgonine and ecgonine methyl ester. The use of the identification point (IP) system in spiked, proficiency testing samples and certified reference materials provided acceptable results in all cases while in some cases real positive samples did not provide acceptable results. Failure to meet the EU criteria was attributed to low fragmentation at the lower concentrations and the ion suppression effect while both factors affected compliance with the IP system. The identification of cocaine and its metabolites was considerably improved by using a combination of ammonium formate and formic acid as the LC mobile phase. It appears that poor in-source fragmentation in single quadrupole LC/MS and ion suppression may constitute a problem with drug identification when implementing the IP system in real samples, resulting in false negative results. Further investigation is needed for the use of such IP systems to be suitable for use in LC/MS methods. PMID- 19902419 TI - Real-time trace detection and identification of chemical warfare agent simulants using recent advances in proton transfer reaction time-of-flight mass spectrometry. AB - This work demonstrates for the first time the potential of using recent developments in proton transfer reaction mass spectrometry for the rapid detection and identification of chemical warfare agents (CWAs) in real-time. A high-resolution (m/Deltam up to 8000) and high-sensitivity (approximately 50 cps/ppbv) proton transfer reaction time-of-flight mass spectrometer (PTR-TOF 8000 from Ionicon Analytik GmBH) has been successfully used to detect a number of CWA simulants at room temperature; namely dimethyl methylphosphonate, diethyl methylphosphonate, diisopropyl methylphosphonate, dipropylene glycol monomethyl ether and 2-chloroethyl ethyl sulfide. Importantly, we demonstrate in this paper the potential to identify CWAs with a high level of confidence in complex chemical environments, where multiple threat agents and interferents could also be present in trace amounts, thereby reducing the risk of false positives. Instantaneous detection and identification of trace quantities of chemical threats using proton transfer reaction mass spectrometry could form the basis for a timely warning system capability with greater precision and accuracy than is currently provided by existing analytical technologies. PMID- 19902420 TI - Reverse thin layer method for enhanced ion yield of oligosaccharides in matrix assisted laser desorption/ionization. AB - A sample preparation method that is suitable for sensitive detection of underivatized oligosaccharides by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has been investigated. As compared with the conventional dried-droplet or ethanol (EtOH) recrystallization method, superior mass spectra in terms of ion yield and signal-to-noise (s/n) ratio were obtained when methanol (MeOH) was used as a solvent for the mixture of matrix and oligosaccharides. Based on these results, a new sample preparation method, named the 'reverse thin layer method', was developed. This method comprises two steps: first, complete drying of the oligosaccharide solution on the MALDI target plate; and second, deposition of the matrix dissolved in a small amount of MeOH. Using this method, a relatively homogeneous matrix crystal was generated and higher yields of both positive and negative ions were obtained from oligosaccharides compared with conventional methods. Notably, the method can be applied to various matrices including both solid and liquid matrices. PMID- 19902421 TI - Analysis of phenolic glycosides from Ilex litseaefolia using electrospray ionization quadrupole time-of-flight mass spectrometry. PMID- 19902422 TI - Selective photocatalytic degradation of poly(ethylene glycol) additives using TiO2 surface-assisted laser desorption/ionization mass spectrometry. PMID- 19902423 TI - The successful inclusion of succinylacetone as a marker of tyrosinemia type I in Tuscany newborn screening program. PMID- 19902424 TI - Lamivudine maintenance beyond one year after HBeAg seroconversion is a major factor for sustained virologic response in HBeAg-positive chronic hepatitis B. AB - The reported durability of virologic response after successful lamivudine monotherapy is variable, and the question remains as to whether virologic responses can be maintained over an extended follow-up period. The aim of this study was to investigate posttreatment durability, the optimal duration of additional treatment after HBeAg clearance or seroconversion, and determinants for sustained virologic response (SVR) following lamivudine monotherapy in patients with HBeAg-positive chronic hepatitis B (CHB). From January 1999 to August 2004, 178 Korean patients with HBeAg-positive CHB were treated with lamivudine and achieved complete responses, defined as a loss of serum HBeAg and hepatitis B virus DNA, and alanine aminotransferase normalization. The mean duration of lamivudine monotherapy was 26 months (range, 12-77). SVR was maintained in 138 patients (77.5%). Host and viral factors were compared between 138 patients with SVR and 40 patients whose response was not sustained. The cumulative relapse rates increased from 15.9% at 1 year to 30.2% at 5 years, with a mean time to relapse after cessation of lamivudine of 12 months (range, 7-42). Most relapses occurred within 2 years after discontinuation of lamivudine (33/40, 82.5%). On multivariate analysis, age 59). Individuals with this genotype were found to have the lowest PON1 enzyme activities among the -107T/C genotypes. Triple combined haplotype QRLMTC was found to be 6.94- and 10.4-times protective against ischemic stroke in the overall and the elderly population, respectively. 55LL genotype was associated with a 1.78-fold increase in the risk of ischemic stroke. CONCLUSION: PON1 genotypes, but not activities, are related with the risk of stroke. PMID- 19902426 TI - Improved preparation and detection of cytochrome P450 isoforms using MS methods. AB - Cytochromes P450 (CYPs) are a superfamily of mixed function oxidases, which in the liver have great significance to the pharmaceutical industry because their expression will determine the fate of most clinical agents. CYPs are also targets for inhibitors of hormone-dependent diseases and conversion of prodrugs to active agents in normal and cancer tissues. We have applied simple modifications to established methods of isolating CYPs, using 8 M urea to solubilise microsomal proteins and specific molecular weight gel bands for in-gel digestion in combination with nanoHPLC MALDI MS to acquire peptide MS/MS spectra for database searching. As a consequence of the changes we significantly improved the yield of proteomic data, identifying 26 mouse CYPs (CYP1a2, 2a4, 2a5, 2a12, 2b9, 2c29, 2c37, 2c39, 2c40, 2c50, 2c54, 2c70, 2d9, 2d10, 2d26, 2e1, 2f2, 2j5, 3a11, 3a13, 3a25, 3a41, 4a14, 4f14, 8b1 and 27a1) with an average sequence coverage of 30.1%, including some previously undetected highly homologous isoforms. In addition, other important enzymes in drug metabolism are also identified. There is a divergence of opinion over the expression of CYP1a1 in liver and we could not detect the presence of this isoform. In order to provide definitive evidence of the ability to detect CYP1a1, we analysed CHO cells transfected with human CYP1A1 and identified unique peptides that differentiated this isoform from human CYP1A2. PMID- 19902427 TI - A new generation of cross-linkers for selective detection by MALDI MS. AB - We designed a new cross-linker bearing a CHCA moiety. The use of the CHCA-tagged cross-linker JMV 3378 in conjunction with a neutral MALDI matrix alpha-cyano-4 hydroxycinnamic methyl ester enabled specific signal enhancement in MALDI-TOF MS of cross-link containing peptides. Discrimination between modified and non modified peptides can be achieved by comparison of two spectra, one using CHCA and the other using the alpha-cyano-4-hydroxycinnamic methyl ester matrix. The methodology was validated using cytochrome c and apo-myoglobine as model proteins. PMID- 19902428 TI - Symbol nomenclature for glycan representation. AB - The glycan symbol nomenclature proposed by Harvey et al. in these pages has relative advantages and disadvantages. The use of symbols to depict glycans originated from Kornfeld in 1978, was systematized in the First Edition of "Essentials of Glycobiology" and updated for the second edition, with input from relevant organizations such as the Consortium for Functional Glycomics. We also note that >200 illustrations in the second edition have already been published using our nomenclature and are available for download at PubMed. PMID- 19902429 TI - Galectin-9 expands immunosuppressive macrophages to ameliorate T-cell-mediated lung inflammation. AB - Galectin-9 (Gal-9) plays pivotal roles in the modulation of innate and adaptive immunity to suppress T-cell-mediated autoimmune models. However, it remains unclear if Gal-9 plays a suppressive role for T-cell function in non-autoimmune disease models. We assessed the effects of Gal-9 on experimental hypersensitivity pneumonitis induced by Trichosporon asahii. When Gal-9 was given subcutaneously to C57BL/6 mice at the time of challenge with T. asahii, it significantly suppressed T. asahii-induced lung inflammation, as the levels of IL-1, IL-6, IFN gamma, and IL-17 were significantly reduced in the BALF of Gal-9-treated mice. Moreover, co-culture of anti-CD3-stimulated CD4 T cells with BALF cells harvested from Gal-9-treated mice on day 1 resulted in diminished CD4 T-cell proliferation and decreased levels of IFN-gamma and IL-17. CD11b(+)Ly-6C(high)F4/80(+) BALF Mphi expanded by Gal-9 were responsible for the suppression. We further found in vitro that Gal-9, only in the presence of T. asahii, expands CD11b(+)Ly 6C(high)F4/80(+) cells from BM cells, and the cells suppress T-cell proliferation and IFN-gamma and IL-17 production. The present results indicate that Gal-9 expands immunosuppressive CD11b(+)Ly-6C(high) Mphi to ameliorate Th1/Th17 cell mediated hypersensitivity pneumonitis. PMID- 19902430 TI - VEGFR2 is selectively expressed by FOXP3high CD4+ Treg. AB - CD25+ FOXP3+CD4+ T cells (Treg) have been considered to play an important role in immune tolerance against several tumor antigens. It has also been indicated that high-level expression of FOXP3 (FOXP3high) is sufficient to confer suppressive activity to normal non-Treg. Here, we showed for the first time that vascular endothelial growth factor receptor 2 (VEGFR2) is selectively expressed by FOXP3high but not FOXP3low Treg. Such VEGFR2+ Treg exist in several tissues including PBMC and malignant effusion-derived lymphocytes. In conclusion, VEGFR2 may be a novel target for controlling Treg with highly suppressive function. PMID- 19902431 TI - Enrichment of (8,4) single-walled carbon nanotubes through coextraction with heparin. AB - Heparin sodium salt is investigated as a dispersant for dispersing single-walled carbon nanotubes (SWNTs). Photoluminescence excitation (PLE) spectroscopy is used for identification and abundance estimation of the chiral species. It is found that heparin sodium salt preferentially disperses larger-diameter Hipco SWNTs. When used to disperse CoMoCAT nanotube samples, heparin has a strong preference for (8,4) tubes, which have larger diameter than the predominant (6,5) in pristine CoMoCAT samples. PLE intensity due to (8,4) tubes increases from 7% to 60% of the total after threefold extractions. Computer modeling verifies that the complex of (8,4) SWNTs and heparin has the lowest binding energy amongst the four semiconducting species present in CoMoCAT. Network field-effect transistors are successfully made with CoMoCAT/heparin and CoMoCAT/sodium dodecylbenzene sulfonate (SDBS)-heparin (x3), confirming the easy removability of heparin. PMID- 19902432 TI - Tuning array morphology for high-strength carbon-nanotube fibers. AB - Vertically aligned carbon-nanotube arrays are synthesized by chemical vapor deposition. Carbon-nanotube fibers are directly spun from the obtained nanotube arrays and then tested mechanically. A strong correlation between the array morphologies and the mechanical properties of the fibers is observed: well aligned arrays yield fibers with much higher performance, while wavy and entangled arrays give poor fiber properties. More importantly, such array morphologies could be controlled by introducing hydrogen or oxygen during the nanotube synthesis. By simply switching the growth condition from 150 ppm oxygen addition to 2% hydrogen addition, the nanotube array changes from the wavy morphology to the well-aligned morphology, and correspondingly the tensile strength of the resultant fibers could be increased by 4.5 times, from 0.29 GPa for the fibers spun from the oxygen-assistance-grown nanotube arrays to 1.3 GPa for the fibers spun from the hydrogen-assistance-grown nanotube arrays. The detailed effects of hydrogen and oxygen on the nanotube growth, especially on the growth rate and the array spinnability, are extensively studied. The formation mechanism of the different morphologies of the nanotube arrays and the failure mechanism of the nanotube fibers are also discussed in detail. PMID- 19902433 TI - Tunable two-dimensional binary molecular networks. AB - A novel approach to constructing tunable and robust 2D binary molecular nanostructures on an inert graphite surface is presented. The guest molecules are embedded into a host molecular matrix and constrained via the formation of multiple intermolecular hydrogen bonds. By varying the binary molecular ratio and the molecular geometry, various molecular arrays with tunable intermolecular distances are fabricated. The results suggest a promising route for the fabrication of ordered and stable molecular nanostructure arrays for molecular sensors, molecular spintronic devices, and molecular p-n nanojunctions. PMID- 19902434 TI - Efficient and stable solid-state dye-sensitized solar cells based on a high-molar extinction-coefficient sensitizer. AB - The high-molar-extinction-coefficient heteroleptic ruthenium dye, cis-Ru (4,4' bis(5-octylthieno[3,2-b] thiophen-2-yl)-2,2'-bipyridine) (4,4'-dicarboxyl-2,2' bipyridine) (NCS)(2), exhibits an AM 1.5 solar (100 mW cm(-2))-to-electric power conversion efficiency of 4.6% in a solid-state dye-sensitized solar cell (SSDSC) with 2,2', 7,7'-tetrakis-(N,N-di-p-methoxyphenylamine)9,9'-spirobifluorene (spiro MeOTAD) as the organic hole-transporting material. These SSDSC devices exhibit good durability during accelerated tests under visible-light soaking for 1000 h at 60 degrees C. This demonstration elucidates a class of photovoltaic devices with potential for stable and low-cost power generation. The electron recombination dynamics and charge collection that take place at the dye sensitized heterojunction are studied by means of impedance and transient photovoltage decay techniques. PMID- 19902435 TI - Ionic liquid-based routes to conversion or reuse of recycled ammonium perchlorate. AB - New, potentially green, and efficient synthetic routes for the remediation and/or re-use of perchlorate-based energetic materials have been developed. Four simple organic imidazolium- and phosphonium-based perchlorate salts/ionic liquids have been synthesized by simple, inexpensive, and nonhazardous methods, using ammonium perchlorate as the perchlorate source. By appropriate choice of the cation, perchlorate can be incorporated into an ionic liquid which serves as its own electrolyte for the electrochemical reduction of the perchlorate anion, allowing for the regeneration of the chloride-based parent ionic liquid. The electrochemical degradation of the hazardous perchlorate ion and its conversion to harmless chloride during electrolysis was studied using IR and (35)Cl NMR spectroscopies. PMID- 19902436 TI - Hydration of sugars in the gas phase: regioselectivity and conformational choice in N-acetyl glucosamine and glucose. AB - The influence of an acetamido group in directing the preferred choice of hydration sites in glucosamine and a consequent extension of the working rules governing regioselective hydration and conformational choice, have been revealed through comparisons between the conformations and structures of "free" and multiply hydrated phenyl N-acetyl-beta-D-glucosamine (betapGlcNAc) and phenyl beta-D-glucopyranoside (betapGlc), isolated in the gas phase at low temperatures. The structures have been assigned through infrared ion depletion spectroscopy conducted in a supersonic jet expansion, coupled with computational methods. The acetamido motif provides a hydration focus that overwhelms the directing role of the hydroxymethyl group; in multiply hydrated betapGlcNAc the water molecules are all located around the acetamido motif, on the "axial" faces of the pyranose ring rather than around its edge, despite the equatorial disposition of all the hydrophilic groups in the ring. The striking and unprecedented role of the C-2 acetamido group in controlling hydration structures may, in part, explain the differing and widespread roles of GlcNAc, and perhaps GalNAc, in nature. PMID- 19902437 TI - Order versus disorder in chiral tetrathiafulvalene-oxazoline radical-cation salts: structural and theoretical investigations and physical properties. AB - Electrocrystallization experiments with the chiral ethylenedithio tetrathiafulvalene-methyl-oxazoline (EDT-TTF-OX) donors (R)-, (S)-, and (rac)-1 have provided two series of mixed-valence salts with the PF(6) (-) and [Au(CN)(2)](-) anions. Within each series the cell parameters are the same for the three R, S, and rac compounds, except for the space group, which is centrosymmetric triclinic P$?bar 1$ for the racemic forms and noncentrosymmetric P1 for the enantiopure salts. In the racemic salt [(rac)-1](2)PF(6) the two enantiomers crystallize disordered on the same crystallographic site with a site occupational factor of 0.6:0.4, whereas this type of disorder is not possible in the enantiopure salts. Both s-cis and s-trans conformations, when taking into account the mutual orientation of the TTF and oxazoline moieties, are present in this first series. In sharp contrast, in the series of salts [1](2)[Au(CN)(2)], only the s-trans conformation is observed with no structural disorder. Theoretical calculations at the DFT level of theory revealed a very small energy difference between the two stable planar s-cis and s-trans conformations, which are both energy minima in either neutral or oxidized states. Single-crystal conductivity measurements showed metallic-like behavior for all the salts down to 220-250 K with a smooth increase in resistivity at lower temperatures. The conductivity at room temperature is 5 S cm(-1) for [(rac)-1](2)PF(6), in which disorder was observed, whereas for [(R)-1](2)PF(6) and [(S)-1](2)PF(6) the average value is around 100 S cm(-1). In the second series of salts the conductivity at room temperature is 125-130 S cm(-1) for [(rac)-1](2)[Au(CN)(2)] and [(R)-1](2)[Au(CN)(2)]. Extended Huckel band structure calculations revealed identical features for the three salts of the [1](2)[Au(CN)(2)] series and are consistent with the electronic structures of quasi-one-dimensional conductors. PMID- 19902438 TI - Noncovalent synthesis of hierarchical zinc phosphates from a single Zn(4)O(12)P(4) double-four-ring building block: dimensionality control through the choice of auxiliary ligands. AB - In contrast to the well-known reaction of phosphonic acids RP(O)(OH)(2) with divalent transition-metal ions that yields layered metal phosphonates [RPO(3)M(H(2)O)](n), the 2,6-diisopropylphenyl ester of phosphoric acid, dippH(2), reacts with zinc acetate in methanol under ambient conditions to afford tetrameric zinc phosphate [(ArO)PO(3)Zn(MeOH)](4) (1). The coordinated methanol in 1 can be readily exchanged by stronger Lewis basic ligands at room temperature. This strategy opens up a new avenue for building double-four-ring (D4R) cubane-based supramolecular assemblies through strong intercubane hydrogen bonding interactions. Seventeen pyridinic ligands have been used to synthesize as many D4R cubanes [(ArO)PO(3)Zn(L)](4) (2-18) from 1. The ligands have been chosen in such a way that the majority of them contain an additional functional group that could be used for noncovalent synthesis of extended structures. When the ligand does not contain any other hydrogen-bonding donor-acceptor sites (e.g., 2,4,6-trimethylpyridine (collidine)), zero-dimensional D4R cubanes have been obtained. The use of pyridine, lutidine, 2-aminopyridine, and 2,6 diaminopyridine, however, results in the formation of linear or zigzag one dimensional assemblies of D4R cubanes through strong intermolecular C-H...O or N H...O interactions. Construction of two-dimensional assemblies of zinc phosphates has been achieved by employing 2-hydroxypyridine or 2-methylimidazole as the exo cubane ligand on zinc centers. The introduction of an alcohol side chain on the pyridinic ligand in such a way that the -CH(2)OH group cannot participate in intracubane hydrogen bonding (e.g., pyridine-3-methanol, pyridine-4-methanol, and 3,5-dimethylpyrazole-N-ethanol) leads to the facile noncovalent synthesis of three-dimensional framework structures. Apart from being useful as building blocks for noncovalent synthesis of zeolite-like materials, compounds 1-18 can also be thermolyzed at approximately 500 degrees C to yield high-purity zinc pyrophosphate (Zn(2)P(2)O(7)) ceramic material. PMID- 19902439 TI - Metallocyclo- and polyphosphazenes containing gold or silver: thermolytic transformation into nanostructured materials. AB - A cyclotriphosphazene bearing two 4-oxypyridine groups on the same phosphorus atom, gem-[N(3)P(3)(O(2)C(12)H(8))(2)(OC(5)H(4)N-4)(2)] (I), and its analogous polymer [{NP(O(2)C(12)H(8))}(0.7){NP(OC(5)H(4)N-4)(2)}(0.3)](n) (II), have been used to prepare gold or silver, cyclic and polymeric, metallophosphazenes. The following complexes, gem-[N(3)P(3)(O(2)C(12)H(8))(2)(OC(5)H(4)N-4{ML})(2)] (ML=Au(C(6)F(5)) (1) or Au(C(6)F(5))(3) (2)), [N(3)P(3)(O(2)C(12)H(8))(2)(OC(5)H(4)N-4{AuPPh(3)})(2)][NO(3)](2) (3), and [N(3)P(3)(O(2)C(12)H(8))(2)(OC(5)H(4)N-4{AgPPh(2)R})(2)][SO(3)CF(3)](2) (R=Ph (4) or Me (5)) have been obtained. Complexes 1 and 4 are excellent models for the preparation of the analogous polymers [{NP(O(2)C(12)H(8))}(0.7){NP(OC(5)H(4)N 4{ML})(2)}(0.3)](n) (ML=Au(C(6)F(5)) (P1), Ag(OSO(2)CF(3))PPh(3) (P2)). All complexes have been characterized by elemental analysis, various spectroscopic methods, and mass spectrometry. The polymers were further investigated by thermochemical methods (thermogravimetric analysis) and differential scanning calorimetry. For compounds 1-5 and for the starting phosphazene I, a mixture of different stereoisomers may be expected. The stereochemistry in solution has been studied by variable-temperature NMR spectroscopy studies, which provided evidence for interconversion processes that involve changes in the chirality of a 2,2' dioxybiphenyl group. A single-crystal X-ray analysis of the gold complex 2 confirmed not only the proposed structure, but also S,S and R,R configurations at the two biphenoxy-substituted phosphorus centers, in contrast to those observed for the precursor I. Pyrolysis of these new metallophosphazenes was also studied. Notably, pyrolysis of the gold derivatives gave macroporous metallic gold sponges without the requirement of either an external reducing agent or a porous support. These materials were all characterized by XRD, TEM, SEM, and energy-dispersive X ray spectroscopy. PMID- 19902440 TI - A 3D copper(II) coordination framework showing different kinetic and thermodynamic crystal transformations through removal of guest water cubes. PMID- 19902441 TI - Heparin-coated gold nanoparticles for liver-specific CT imaging. PMID- 19902442 TI - Encumbered dispiro[fluorene-indenofluorene]: mechanistic insights. PMID- 19902443 TI - Oxidative coupling of diaryldisulfides by MoCl5 to thianthrenes. PMID- 19902444 TI - Sequential and modular synthesis of chiral 1,3-diols with two stereogenic centers: access to all four stereoisomers by combination of organo- and biocatalysis. PMID- 19902445 TI - Highly efficient "grafting onto" a polypeptide backbone using click chemistry. PMID- 19902446 TI - Copper as a powerful catalyst in the direct alkynylation of azoles. PMID- 19902447 TI - A dialdehyde cyclization cascade: an approach to pleuromutilin. PMID- 19902448 TI - Synthesis and pharmacological evaluation of dimeric follicle-stimulating hormone receptor antagonists. AB - A series of homo- and heterodimeric compounds encompassing the follicle stimulating hormone receptor (FSHR) antagonist (R)-1 and its inactive conformer (S)-1 connected through ethylene glycol spacers of various lengths is described. Evaluation of these compounds reveals that dimeric compounds, with a spacer of sufficient length, bearing two active copies of the antagonist are more potent relative to dimeric compounds in which one of the active pharmacophores is replaced by an inactive conformer. Interestingly, the opposite trend is observed if a short spacer is used, indicating that these compounds may be valuable tools to study FSHR dimerization in greater detail. PMID- 19902449 TI - Remote sensitized photoisomerization via through-bond triplet-triplet energy transfer mediated by a salt bridge in a supramolecular dyad. AB - A supramolecular dyad, BP-(amidinium-carboxylate)-NBD is constructed, in which benzophenone (BP) and norbornadiene (NBD) are connected via an amidinium carboxylate salt bridge. The photophysical and photochemical properties of the assembled BP-(amidinium-carboxylate)-NBD dyad are examined. The phosphorescence of the BP chromophore is efficiently quenched by the NBD group in BP-(amidinium carboxylate)-NBD via the salt bridge. Time-resolved spectroscopy measurements indicate that the lifetime of the BP triplet state in BP-(amidinium-carboxylate) NBD is shortened due to the quenching by the NBD group. Selective excitation of the BP chromophore results in isomerization of the NBD group to quadricyclane (QC). All of these observations suggest that the triplet-triplet energy transfer occurs efficiently in the BP-(amidinium-carboxylate)-NBD salt bridge system. The triplet-triplet energy transfer process proceeds with efficiencies of approximately 0.87, 0.98 and the rate constants 1.8x10(3) s(-1), and 1.3x10(7) s( 1) at 77 K and room temperature, respectively. The mechanism for the triplet triplet energy transfer is proposed to proceed via a "through-bond" electron exchange process, and the non-covalent bonds amidinium-carboxylate salt bridge can mediate the triplet-triplet energy transfer process effectively for photochemical conversion. PMID- 19902450 TI - A sub-50-nm monosized superparamagnetic Fe3O4@SiO2 T2-weighted MRI contrast agent: highly reproducible synthesis of uniform single-loaded core-shell nanostructures. AB - Oleic acid stabilized superparamagnetic iron oxide nanoparticles (SPION) were selected as the cores for fabrication of sub-50-nm monodisperse single-loaded SPION@SiO2 core-shell nanostructures. Parameters that influence the formation of SPION@SiO2 in the water-in-oil reverse microemulsion system have been systematically investigated. The sufficiently high concentration of well dispersed SPION, together with an appropriately low injection rate of tetraethoxysilane, were found to be the keys to efficiently prevent the homogeneous nucleation of silica and obtain a high-quality single-loaded core shell nanocomposite. A more detailed mechanism for incorporating oleic acid capped inorganic functional nanoparticles into silica is proposed on the basis of previous reports and our new experimental results. Finally, the as-synthesized SPION@SiO2 nanospheres are exploited as an MRI-enhanced contrast agent, and their contrast effect in solution is tested by using a clinical MRI instrument. PMID- 19902451 TI - Brief communication: Forelimb compliance in arboreal and terrestrial opossums. AB - Primates display high forelimb compliance (increased elbow joint yield) compared to most other mammals. Forelimb compliance, which is especially marked among arboreal primates, moderates vertical oscillations of the body and peak vertical forces and may represent a basal adaptation of primates for locomotion on thin, flexible branches. However, Larney and Larson (Am J Phys Anthropol 125 [2004] 42 50) reported that marsupials have forelimb compliance comparable to or greater than that of most primates, but did not distinguish between arboreal and terrestrial marsupials. If forelimb compliance is functionally linked to locomotion on thin branches, then elbow yield should be highest in marsupials relying on arboreal substrates more often. To test this hypothesis, we compared forelimb compliance between two didelphid marsupials, Caluromys philander (an arboreal opossum relying heavily on thin branches) and Monodelphis domestica (an opossum that spends most of its time on the ground). Animals were videorecorded while walking on a runway or a horizontal 7-mm pole. Caluromys showed higher elbow yield (greater changes in degrees of elbow flexion) on both substrates, similar to that reported for arboreal primates. Monodelphis was characterized by lower elbow yield that was intermediate between the values reported by Larney and Larson (Am J Phys Anthropol 125 [2004] 42-50) for more terrestrial primates and rodents. This finding adds evidence to a model suggesting a functional link between arboreality--particularly locomotion on thin, flexible branches--and forelimb compliance. These data add another convergent trait between arboreal primates, Caluromys, and other arboreal marsupials and support the argument that all primates evolved from a common ancestor that was a fine-branch arborealist. PMID- 19902452 TI - Spouse selection by health status and physical traits. Sardinia, 1856-1925. AB - Military medical information and data from civil registers of death and marriage have been used to study the role of physical characteristics and health conditions in explaining access to marriage for the male population of Alghero, a small city located in Sardinia Island (Italy), at the turn of 19th century. Literature data about contemporary populations have already demonstrated the influence of somatic traits in the mate choice. The results presented here show that men with low height and poor health status at the age of 20 were negatively selected for marriage. This holds true also in a society where families often arranged marriages for their children. This pattern of male selection on marriage was found to be particularly marked among the richest and wealthiest SES groups. Our hypothesis is that this social group carefully selected for marriage those individuals who were apparently healthier and therefore more likely to guarantee good health status and better life conditions to offspring. In evolutionary terms, the mate choice component of sexual selection suggests that the height of prospective partners could be claimed as one of the determinants, along with other environmental causes, of the observed higher stature of men belonging to the wealthiest social strata of the Alghero population. PMID- 19902453 TI - Gleaning signals about the past from cemetery data. AB - Cemetery headstones provide an easily accessible source of demographic data in human populations. In common with other sources of demographic data, such as skeletal samples, cemetery data may not be representative of the populations from which they were derived. In some circumstances they can be reasonably representative, however, and in such cases they may provide signals about demographic changes in the population that contributed to the cemetery. We present here analyses of burials occurring between 1900 and 1990 at the Columbia Cemetery in Columbia, Missouri. Our analyses, in combination with archival materials relating to infrastructure improvements in Columbia and data on infectious disease mortality in the state of Missouri, show that patterns of death observed in the cemetery data provide evidence for the timing of changes in the health of Columbia's residents. At the time that major improvements in sanitation and hygiene were implemented, burials of individuals dying under age 45 decreased significantly while burials of individuals older than 45 remained relatively high. Furthermore, data on infectious disease mortality indicate significant declines in deaths from water- and milk-borne infections, but no change in mortality from respiratory illnesses. These data also indicate that observed changes occurred about a decade later in Columbia than in large cities and more densely populated states elsewhere in the United States. Thus, this study illustrates the value of cemetery data in helping to fill gaps about how and when different events known to affect patterns of birth and death may have played out across time and space. PMID- 19902454 TI - The influence of masticatory loading on craniofacial morphology: A test case across technological transitions in the Ohio valley. AB - Masticatory loading is one of the main environmental stimuli that generate craniofacial variation among recent humans. Experimental studies on a wide variety of mammals, including those with retrognathic postcanine teeth, predict that responses to masticatory loading will be greater in the occlusal plane, the inferior rostrum, and regions associated with the attachments of the temporalis and masseter muscles. Here we test these experimentally-derived predictions on an extinct human population from the middle and upper Ohio valley that underwent a marked shift from hunting-gathering to extensive farming during the last 3,000 years and for which we have good archaeological evidence about diet and food processing technology. Geometric morphometric methods were used to detect and measure the putative effect of diet changes on cranial shape independent of size. Our results partially confirm only some of the experimental predictions. The effect of softer and/or less tough diets on craniofacial shape seem to be concentrated in the relative reduction of the temporal fossa and in a displacement of the attachment of the temporal muscle. However, there were few differences in craniofacial shape in regions closer to the occlusal plane. These results highlight the utility of exploring specific localized morphological shifts using a hierarchical model of craniofacial integration. PMID- 19902455 TI - The harsh life on the 15th century Croatia-Ottoman empire military border: analyzing and identifying the reasons for the massacre in Cepin. AB - Excavation of the historic period cemetery in Cepin, Croatia revealed the presence of a large number of perimortem injuries distributed among males, females, and subadults. Archaeological and historical data suggest these individuals were victims of a raid carried out by Turkish akinji light cavalry in 1441. Comparisons with the frequencies of perimortem trauma in 12 other, temporally congruent skeletal series from the Balkans (n = 2,123 skeletons) support this assumption. The role of the akinji in the Ottoman army was twofold: to supply war captives, and to terrorize and disperse local populations before the advance of regular troops. This article tests the hypothesis that the purpose of the 1441 raid was the latter. To accomplish this, perimortem trauma in the series were analyzed by sex, age, location, and depth of the injury. A total of 82 perimortem injuries were recorded in 12 males, 7 females, and 3 subadults. The demographic profile of the victims suggests that young adults were specifically targeted in the attack. Significant sex differences are noted in the number, distribution, and pattern of perimortem trauma. Females exhibit significantly more perimortem injuries per individual, and per bone affected, than males. The morphology and pattern of perimortem trauma in females is suggestive of gratuitous violence. Cumulatively, analysis of the osteological data suggest that the objective of the 1441 akinji raid was to spread terror and panic in the Cepin area, either as revenge for recent military setbacks, or as part of a long-term strategy intended to depopulate the area around Osijek. PMID- 19902456 TI - The rediscovery of two Upper Palaeolithic skeletons from Baousso da Torre cave (Liguria-Italy). PMID- 19902457 TI - Height and body mass influence on human body outlines: a quantitative approach using an elliptic Fourier analysis. AB - Many studies use representations of human body outlines to study how individual characteristics, such as height and body mass, affect perception of body shape. These typically involve reality-based stimuli (e.g., pictures) or manipulated stimuli (e.g., drawings). These two classes of stimuli have important drawbacks that limit result interpretations. Realistic stimuli vary in terms of traits that are correlated, which makes it impossible to assess the effect of a single trait independently. In addition, manipulated stimuli usually do not represent realistic morphologies. We describe and examine a method based on elliptic Fourier descriptors to automatically predict and represent body outlines for a given set of predicted variables (e.g., sex, height, and body mass). We first estimate whether these predictive variables are significantly related to human outlines. We find that height and body mass significantly influence body shape. Unlike height, the effect of body mass on shape differs between sexes. Then, we show that we can easily build a regression model that creates hypothetical outlines for an arbitrary set of covariates. These statistically computed outlines are quite realistic and may be used as stimuli in future studies. PMID- 19902458 TI - On the relationship of dental microwear to dental macrowear. AB - Dental microwear analysts have demonstrated that hard diets leave numerous microscopic pits on occlusal surfaces. The relationship between occlusal pitting and gross macrowear, however, is not well known. The current study seeks to elucidate the relationship between dental microwear and macrowear by determining if microscopically pitted teeth are associated with greater expressions of macrowear. This study examined microwear and macrowear on mandibular second molars from 60 prehistoric adult Native Americans representing three dietary regimes (foraging, mixed economy, and agriculture). Initially, two dental microwear feature variables were studied: percentage of pits and mean scratch width. Standard macrowear scores ranged from 4 to 40. ANOVAs suggested that neither of the microwear variables was affected by age or sex, but age affected macrowear scores. Because of this, the sample had a balanced number of young and old adults (i.e., those below and above skeletal age 35). A Pearson's correlation showed no covariation between scratch width and the percentage of pits. Regression analysis indicated that macrowear was not a function of the percentage of pits. However, a significant positive relationship was found between dental macrowear and scratch width. A post priori test found a significant negative relationship between macrowear and the total number of scratches. It is concluded, then, that wide scratches remove more enamel and dentin than do numerous pits, although both cause dental wear. It is suggested here that the term "abrasive" be used to describe those microwear profiles that lead to heavy macrowear and have relatively wide scratches. PMID- 19902459 TI - Mammographic density and the risk of breast cancer recurrence after breast conserving surgery. AB - BACKGROUND: Women with invasive breast cancer who are treated with breast conserving surgery and radiotherapy face a cumulative risk of local disease recurrence of approximately 10% at 10 years. To the authors' knowledge, the role of mammographic density as a risk factor for the development of local recurrence has not been thoroughly evaluated to date. METHODS: Medical records were reviewed for 335 patients who underwent breast-conserving surgery for invasive breast cancer and for whom a pretreatment mammogram was available. Information was recorded concerning mammographic density as well as tumor features, patient characteristics, and adjuvant treatments received. Patients were categorized for mammographic density based on the Wolfe classification as either low (<25% density), intermediate (25-50% density), or high (>50% density). A multivariate survival analysis was conducted using the Cox proportional hazards model with local disease recurrence as the primary endpoint. RESULTS: Patients in the high mammographic density group experienced a much greater risk of local disease recurrence compared with women with the least dense breasts (10-year actuarial risks: 21% vs 5%; hazards ratio [HR], 5.7 [95% confidence interval, 1.6-20; P=.006]). The difference in the rates of disease recurrence at 10 years was pronounced for women who did not receive radiotherapy (40% vs 0% for patients with >50% density and <25% density, respectively; P<.0001). CONCLUSIONS: Mammographic breast density is an important risk factor for local breast cancer recurrence among women not receiving breast irradiation. Mammographic density should be taken into consideration when stratifying patients for clinical trials of partial breast radiotherapy. If confirmed, mammographic density might be used to help determine which patients might benefit from radiotherapy. PMID- 19902460 TI - Relationship of protozoan biomass to phosphate and nitrate removal from activated sludge mixed liquor. AB - The relationship between protozoan biomass concentration and phosphate and nitrate removal was investigated in mixed liquor using three different carbon sources as supplements. The study was carried out using three respective initial biomass concentrations in a shaking flask environment. Samples were taken every 24 h to determine phosphate, nitrate, dissolved oxygen and chemical oxygen demand. The results revealed a direct relationship between decreases in nutrient concentrations and increases in cell densities of the isolates. Between 24 and 96 h, the increases in the protozoan density corresponded to a phosphate decreases from initial ranges of 55.42-57.36 mg/L, 50.27-51.17 mg/L and 50.01-50.83 mg/L to final ranges of 2.46-11.90 mg/L, 0.61-11.80 mg/L and 1.29-13.89 mg/L, in the presence of Aspidisca, Trachelophyllum and Peranema, respectively. Nitrate concentrations were observed to decrease from initial ranges of 23.84-25.90 mg/L, 23.94-25.84 mg/L and 26.12-26.54 mg/L to final ranges of 0.11-6.32 mg/L, 0.16 5.60 mg/L and 0.24-9.04 mg/L, respectively. The study had revealed that an increase in cell density of the test isolates produces a corresponding increase in phosphate and nitrate removal. PMID- 19902461 TI - Recent trends and some applications of isothermal titration calorimetry in biotechnology. AB - Isothermal titration calorimeters (ITCs) are thermodynamic instruments used for the determination of enthalpy changes in any physical/chemical reaction. This can be applied in various fields of biotechnology. This review explains ITC applications, especially in bioseparation, drug development and cell metabolism. In liquid chromatography, the separation/purification of specific proteins or polypeptides in a mixture is usually achieved by varying the adsorption affinities of the different proteins/polypeptides for the adsorbent under different mobile-phase conditions and temperatures. Using ITC analysis, the binding mechanism of proteins with adsorbent solid material is derived by elucidating enthalpy and entropy changes, which offer valuable guidelines for designing experimental conditions in chromatographic separation. The binding affinity of a drug with its target is studied by deriving binding enthalpy and binding entropy. To improve the binding affinity, suitable lead compounds for a drug can be identified and their affinity tested by ITC. Recently ITC has also been used in studying cell metabolism. The heat produced by animal cells in culture can be used as a primary indicator of the kinetics of cell metabolism, which provides key information for drug bioactivity and operation parameters for process cell culture. PMID- 19902462 TI - Enhancing clinical trial design of interventions for posttraumatic stress disorder. AB - The 2008 Institute of Medicine review of interventions research for posttraumatic stress disorder (PTSD) concluded that new, well-designed studies are needed to evaluate the efficacy of treatments for PTSD. The Department of Veterans Affairs (VA), the Department of Defense, and the National Institute of Mental Health convened a meeting on research methodology and the VA issued recommendations for design and analysis of randomized controlled clinical trials (RCTs) for PTSD. The rationale that formed the basis for several of the components of the recommendations is discussed here. Fundamental goals of RCT design are described. Strategies in design and analysis that contribute to the goals of an RCT and thereby enhance the likelihood of signal detection are considered. PMID- 19902463 TI - The influence of a major disaster on suicide risk in the population. AB - The authors investigated the relationship between the September 11, 2001 terrorist attacks and suicide risk in New York City from 1990 to 2006. The average monthly suicide rate over the study period was 0.56 per 100,000 people. The monthly rate after September 2001 was 0.11 per 100,000 people lower as compared to the rate in the period before. However, the rate of change in suicide was not significantly different before and after the disaster, and regression discontinuity analysis indicated no change at this date. There was no net change in the suicide rate in New York City attributable to this disaster, suggesting that factors other than exposure to traumatic events (e.g., cultural norms, availability of lethal methods) may be key drivers of suicide risk in this context. PMID- 19902464 TI - A diagnostic interview for acute stress disorder for children and adolescents. AB - The goal of this study was to develop a semistructured clinical interview for assessing acute stress disorder (ASD) in youth and test its psychometric properties. Youth (N = 168) with an acute burn or injury were administered the acute stress disorder module of the Diagnostic Interview for Children and Adolescents (DICA-ASD). The DICA-ASD demonstrated strong psychometric properties, including high internal consistency (alpha = .97) and perfect diagnostic interrater agreement (kappa = 1.00). Participants diagnosed with ASD scored significantly higher than those not diagnosed on validated traumatic stress symptomatology measures but not on other symptomatology measures, providing evidence of convergent and discriminant validity. Preliminary evidence supports the reliability and validity of the first semistructured clinical interview for diagnosing ASD in youth. PMID- 19902465 TI - Establishment and characterization of an androgen receptor-dependent, androgen independent human prostate cancer cell line, LNCaP-CS10. AB - BACKGROUND: Hormone refractoriness is a lethal event for advanced prostate cancer patients, but the mechanisms of the disease are not well elucidated, especially for the so-called "outlaw" pathways of androgen receptor (AR)-dependent, androgen independent hormone-refractory prostate cancer. METHODS: Androgen-dependent prostate cancer LNCaP cells were treated with bicalutamide under an androgen depleted condition to obtain refractory cells. In the obtained cell line, LNCaP CS10, we analyzed the effects of androgen and bicalutamide on cell growth and prostate-specific antigen (PSA) production. In addition, AR gene mutation, AR expression levels, and AR subcellular localizations were analyzed. RESULTS: In LNCaP-CS10, cell growth and PSA production were found under an androgen-depleted condition and were induced by both R1881 and bicalutamide. Knocking down AR by siRNAs did suppress the growth and PSA production of LNCaP-CS10 cells in the androgen-depleted condition. In comparison to LNCaP, amplification or additional new mutations were not found in the AR genes, but AR nuclear translocation induced by bicalutamide was identified in the LNCaP-CS10 cells. The growth and PSA production of xenografted LNCaP-CS10 tumors, which secrete PSA not only in non-castrated SCID mice but also in castrated SCID mice, were not inhibited by bicalutamide. CONCLUSIONS: We have generated a bicalutamide-resistant and androgen-independent prostate cancer cell line, LNCaP-CS10, with outlaw activation both in vitro and in vivo. The LNCaP-CS10 cell line is beneficial for elucidating outlaw pathway mechanisms and evaluating the efficacy of new therapeutics for hormone-refractory prostate cancer. PMID- 19902466 TI - A functional polymorphism in Pre-miR-146a gene is associated with prostate cancer risk and mature miR-146a expression in vivo. AB - BACKGROUND: A G > C polymorphism (rs2910164) which is located in the sequence of miR-146a precursor, results in a change from a G:U pair to a C:U mismatch in its stem region. To explore whether rs2910164 plays any role in prostate cancer (CaP), we analyzed the association between miR-146a polymorphism and risk of CaP and the expression of miR-146a with different genotypes in CaP tissues in southern Chinese Han population. MATERIALS AND METHODS: Two hundred fifty-one CaP and 280 control subjects were included in the cancer association study, and 15 CaP tissue samples were used to test the expression of the miRNA precursors by real-time quantitative reverse transcription PCR. RESULTS: We found that subjects carrying CC homozygotes had a 0.65-fold reduced risk (95% CI = 0.43-0.99) than those carrying GG/GC genotypes (P = 0.03), and the C allele displayed a lower prevalence of CaP compared with the G allele (OR = 0.73, 95% CI = 0.57-0.94, P = 0.01). Moreover, hsa-miR-146a quantification showed that homozygous carriers of the C-variant had significantly decreased miRNA levels compared to the carriers of the GG/GC genotype. CONCLUSIONS: The natural genetic variation in pre-miR-146a affects the amount of mature miR-146a, contributes to the genetic predisposition to CaP. PMID- 19902467 TI - Chimeric adenoviral vectors incorporating a fiber of human adenovirus 3 efficiently mediate gene transfer into prostate cancer cells. AB - BACKGROUND: We have developed a range of adenoviral (Ad) vectors based on human adenovirus serotype 5 (HAdV-5) displaying the fiber shaft and knob domains of species B viruses (HAdV-3, -11, or -35). These species B Ads utilize different cellular receptors than HAdV-5 for infection. We evaluated whether Ad vectors displaying species B fiber shaft and knob domains (Ad5F3Luc1, Ad5F11Luc1, and Ad5F35Luc1) would efficiently infect cancer cells of distinct origins, including prostate cancer. METHODS: The fiber chimeric Ad vectors were genetically generated and compared with the original Ad vector (Ad5Luc1) for transductional efficiency in a variety of cancer cell lines, including prostate cancer cells and primary prostate epithelial cells (PrEC), using luciferase as a reporter gene. RESULTS: Prostate cancer cell lines infected with Ad5F3Luc1 expressed higher levels of luciferase than Ad5Luc1, as well as the other chimeric Ad vectors. We also analyzed the transductional efficiency via monitoring of luciferase activity in prostate cancer cells when expressed as a fraction of the gene transfer in PrEC cells. In the PC-3 and DU145 cell lines, the gene transfer ratio of cancer cells versus PrEC was once again highest for Ad5F3Luc1. CONCLUSION: Of the investigated chimeric HAdV-5/species B vectors, Ad5F3Luc1 was judged to be the most suitable for targeting prostate cancer cells as it showed the highest transductional efficiency in these cells. It is foreseeable that an Ad vector incorporating the HAdV-3 fiber could potentially be used for prostate cancer gene therapy. PMID- 19902468 TI - Sonographic identification of lower limb venous hypoplasia in the prenatal diagnosis of Klippel-Trenaunay syndrome. AB - We report the prenatal identification of lower-limb venous hypoplasia to support a provisional prenatal diagnosis of Klippel-Trenaunay syndrome (KTS). Ultrasound assessment of a fetus with marked lower-limb edema, cystic areas in the abdomen/pelvis/lower limbs and abnormal development of the feet demonstrated bilateral hypoplasia of the femoral and popliteal veins. The external iliac veins and the great saphenous veins were seen to be normal. The lower limb arterial system was present. These findings supported KTS as the most likely provisional diagnosis, and postnatal clinical evaluation confirmed that the infant is best classified in the spectrum of KTS. Venous hypoplasia was confirmed with a postnatal ultrasound examination of the lower limbs. This case suggests that careful examination of the lower-limb venous system may be helpful in making the prenatal diagnosis of KTS. PMID- 19902469 TI - Ultrasound monitoring in patients undergoing in-vitro fertilization after methotrexate treatment for ectopic pregnancy. AB - OBJECTIVES: To compare sonographic characteristics of the endometrium and follicles during in-vitro fertilization (IVF) before and after methotrexate (MTX) treatment for ectopic pregnancy. METHODS: This retrospective study, conducted at Conception Hospital from January 2000 to July 2007, included all patients diagnosed with an ectopic pregnancy resulting from IVF treatment that was treated with MTX and who then underwent another IVF cycle. We compared the number and size of follicles and the endometrial thickness and quality on the day of human chorionic gonadotropin injection in the cycles before and after the MTX treatment to determine whether MTX had any effect. RESULTS: Eleven patients were included in the study. The median interval between the IVF cycle resulting in ectopic pregnancy and the first IVF cycle after MTX therapy was 180 (range, 150-900) days. There was no statistically significant difference between the before and after MTX treatment groups with respect to number of follicles (14 (3-20) vs. 9 (4-16), P = 0.12), follicle size (16.5 (14.7-21.7) mm vs. 17.8 (14.9-19.8) mm, P = 0.37), endometrial thickness (10.0 (9.5-12.0) mm vs. 10.0 (7.5-14.0) mm, P = 0.31) or endometrial quality (P = 0.32). Four women became pregnant during the IVF cycle following MTX treatment. CONCLUSIONS: Ultrasound monitoring showed no modification of the characteristics of the endometrium or follicles during IVF after MTX treatment for ectopic pregnancy. PMID- 19902470 TI - Immunosuppressive CD14+HLA-DRlow/- monocytes in prostate cancer. AB - OBJECTIVE: To determine if the levels of circulating myeloid-derived suppressor cells increase with progression of prostate cancer (PCa); to determine if such cells could contribute to the relative inefficiency of PCa immunotherapy. MATERIALS AND METHODS: We analyzed peripheral blood mononuclear cells isolated from untreated PCa patients (uPCa; N = 18; mean age +/- SD: 72.1 +/- 6.9 years), tPCa (N = 22; 72.8 +/- 9.8 years) and age matched controls (AMC; N = 12; 68.8 +/- 7.5 years). We quantified surface marker phenotype, differentiation potential, effects on T cell proliferation and intracellular cytokines. RESULTS: We observed an unexpectedly high percentage of a type of myeloid-derived suppressor cells, CD14(+)HLA-DR(low/-) monocytes, in tPCa (30.7 +/- 15.0% of CD14(+) cells) relative to AMC (4.1 +/- 6.5%, P < 0.0001) and uPCa (10.6 +/- 14.3%, P = 0.0001). The levels of CD14(+) HLA-DR(low/-) cells were significantly correlated with circulating PSA levels and treatment with LHRH-agonist leuprolide in combination with either an antiandrogen or dexamethasone. Monocytes from tPCa inhibited autologous T cell proliferation statistically significantly more effectively than AMC monocytes and were defective in their ability to differentiate into phenotypically mature dendritic cells. Isolated CD14(+)HLA-DR(low/-) cells expressed higher levels of intracellular interleukin-10 and suppressed T cell proliferation more effectively than isolated CD14(+)HLA-DR(+) cells. CONCLUSIONS: This is the first report of CD14(+) cells exhibiting reduced expression of HLA-DR molecules in PCa patients. These cells suppress immune cell function in vitro and, plausibly, in vivo, a finding that must be factored into the design of immunotherapy protocols for PCa patients. PMID- 19902471 TI - Three-dimensional ultrasound imaging for discrimination between benign and malignant endometrium in women with postmenopausal bleeding and sonographic endometrial thickness of at least 4.5 mm. AB - OBJECTIVES: To determine whether endometrial volume or power Doppler indices as measured by three-dimensional (3D) ultrasound imaging can discriminate between benign and malignant endometrium, to compare their diagnostic performance with that of endometrial thickness measurement using two-dimensional (2D) ultrasound examination, and to determine whether power Doppler indices add any diagnostic information to endometrial thickness or volume. METHODS: Sixty-two patients with postmenopausal bleeding and endometrial thickness > or = 4.5 mm underwent transvaginal 2D gray-scale and 3D power Doppler ultrasound examination of the corpus uteri. The endometrial volume was calculated, along with the vascularization index (VI), flow index and vascularization flow index (VFI) in the endometrium and in a 2-mm 'shell' surrounding the endometrium. The 'gold standard' was the histological diagnosis of the endometrium obtained by hysteroscopic resection of focal lesions, dilatation and curettage or hysterectomy. Receiver-operating characteristics (ROC) curves were drawn for all measurements to evaluate their ability to distinguish between benign and malignant endometrium. Multivariate logistic regression analysis was used to create mathematical models to estimate the risk of endometrial malignancy. RESULTS: There were 49 benign and 13 malignant endometria. Endometrial thickness and volume were significantly larger in malignant than in benign endometria, and flow indices in the endometrium and endometrial shell were significantly higher. The area under the ROC curve (AUC) of endometrial thickness was 0.82, that of endometrial volume 0.78, and that of the two best power Doppler variables (VI and VFI in the endometrium) 0.82 and 0.82. The best logistic regression model for predicting malignancy contained the variables endometrial thickness (odds ratio 1.2; 95% CI, 1.04-1.30; P = 0.004) and VI in the endometrial 'shell' (odds ratio 1.1; 95% CI, 1.02-1.23; P = 0.01). Its AUC was 0.86. Using its mathematically optimal risk cut-off value (0.22), the model correctly classified seven more benign cases but two fewer malignant cases than the best endometrial thickness cut-off (11.8 mm). Models containing endometrial volume and flow indices performed less well than did endometrial thickness alone (AUC, 0.79 vs. 0.82). CONCLUSIONS: The diagnostic performance for discrimination between benign and malignant endometrium of 3D ultrasound imaging was not superior to that of endometrial thickness as measured by 2D ultrasound examination, and 3D power Doppler imaging added little to endometrial thickness or volume. PMID- 19902472 TI - Monocyte chemotactic protein-1 (MCP-1/CCL2) is associated with prostatic growth dysregulation and benign prostatic hyperplasia. AB - BACKGROUND: Chronic inflammation is commonly observed in benign prostate hyperplasia (BPH), and prostate tissue often contains increased inflammatory infiltrates, including T cells and macrophages. Cytokines are not only key mediators of inflammation but may also play important roles in the initiation and progression of BPH. METHODS: In order to determine what cytokines might be involved in prostatic enlargement, expressed prostatic secretions (EPS) from ex vivo prostates were analyzed by human cytokine antibody microarray and ELISA. Prostate epithelial cells (PrEC) and prostate stromal cells (PrSC) were used for ELISA, proliferation, and Western blot assays. RESULTS: Monocyte chemotactic protein-1 (MCP-1/CCL2) was one of the most elevated proteins in secretions from large prostate glands. PrSC were found to secrete MCP-1; Western blotting showed that both PrSC and PrEC express the MCP-1 receptor CCR2 which by RT-PCR was the CCR2b isoform. Proliferation assays showed that MCP-1 stimulates the proliferation of PrEC, but not PrSC, and that a specific MCP-1 antagonist (RS102895) suppressed this effect. Conditioned medium from PrSC stimulated the proliferation of PrEC as well, an effect completely inhibited by both RS102895 and a neutralizing anti-MCP-1 monoclonal antibody. The inflammatory cytokines interleukin (IL)-1 beta, interferon-gamma, and IL-2 enhanced the secretion of MCP 1 from PrEC and PrSC. In addition, MCP-1 levels in EPS correlated with mRNA levels of the macrophage marker CD68 in the same secretions. CONCLUSIONS: The cytokine MCP-1, of apparent prostatic stromal cell origin, may play an important role in prostatic enlargement and BPH, and is a candidate biomarker for these pathologic processes. PMID- 19902473 TI - Antagonistic interaction between bicalutamide (Casodex) and radiation in androgen positive prostate cancer LNCaP cells. AB - BACKGROUND: Bicalutamide (Casodex) reportedly improves high-risk prostate cancer survival as an adjuvant treatment following radiotherapy. However, biological data for the interaction between bicalutamide and ionizing radiation in concomitant association are lacking. METHODS: To explore this issue, androgen dependent (LNCaP) and -independent (DU145) human prostate cancer cells were exposed for 48 hr to 20, 40, or 80 microM bicalutamide introduced before (neoadjuvant), during (concomitant), or following (adjuvant) radiation. Growth inhibition and cytotoxicity, cell cycle distribution and expression of the prostate serum antigen (PSA) and androgen receptor (AR), were measured as endpoints. RESULTS: Bicalutamide-induced cytotoxic and cytostatic effects were found to be correlated with a marked G1 phase arrest and S phase depression. The drug down-regulated PSA and AR proteins and psa mRNA in LNCaP cells. However, transient up-regulation of the expression of ar mRNA was observed in the presence of 40 microM drug. DU145 cells did not express PSA and proved to be comparatively resistant to the drug from both cytostatic and cytotoxic effects. Bicalutamide dose-dependently induced a significant decrease of radiation susceptibility among drug survivors in LNCaP cells, whilst the interaction appeared to be additive in DU145 cells. CONCLUSIONS: The antagonistic radiation-drug interaction observed in LNCaP cells is of significance in relation to combined radiotherapy-bicalutamide treatments directed against tumors expressing the AR. The results suggest that bicalutamide is amenable to combined schedule with radiotherapy provided the drug and radiation are not given in close temporal proximity. PMID- 19902474 TI - Replication of prostate cancer risk loci on 8q24, 11q13, 17q12, 19q33, and Xp11 in African Americans. AB - BACKGROUND: Prostate cancer (Pca) is a common malignancy that disproportionately affects African American men (AA). Recently there have been several genome-wide association studies (GWAS) implicating new prostate cancer risk loci along chromosomes 2, 3, 6, 7, 8, 10, 11, 12, 17, 19, and X in populations of European ancestry. Given the higher incidence and mortality for AAs, and differences in allele frequencies and haplotype structures between African and European descent populations, it is important to assess the impact of these candidate risk loci in AAs. METHODS: Here we evaluated 20 single nucleotide polymorphisms (SNPs) associated with prostate cancer risk in recent GWAS studies, in AA prostate cancer cases and controls. RESULTS: We replicated five of the SNPs in our AA population, rs10896449 on 11q13.2 (P = 0.009), rs2735839 on 19q33.33 region, (P = 0.04), rs443076 on chromosome 17q12 (P = 0.008), rs5945572 on Xp11.22 (P = 0.05), as well as the rare variant specific to west African ancestry, bd11934905 in region 2 of 8q24 (P = 1 x 10(-4)). CONCLUSIONS: While we were able to replicate a few of the previous GWAS SNPs, we were not able to confirm the vast majority of these associations in our AA population. This finding further supports the need to perform GWAS and additional fine mapping in AAs to locate additional susceptibility loci. PMID- 19902475 TI - Gel infusion sonography in the evaluation of the uterine cavity. AB - OBJECTIVES: To compare gel infusion sonohysterography (GIS) with saline contrast sonohysterography (SCSH) with regard to technical feasibility and procedure related pain experienced by patients. METHODS: This prospective observational cohort study included 551 consecutive patients with abnormal bleeding: SCSH was attempted in the first 402 women and GIS was attempted in the following 149. All procedures were performed by the same examiner, in the same clinical setting, using a 2-mm diameter catheter. After contrast sonohysterography, most patients underwent office hysteroscopy (n = 502) and endometrial sampling (n = 323). The women were asked to rate the pain experienced during each procedure using a 100 mm visual analog scale (VAS). Patients' characteristics, ultrasound findings, histological diagnosis, technical failures and procedure-related pain were compared between the two procedures. RESULTS: The mean +/- SD VAS score for contrast sonography, subsequent hysteroscopy and endometrial biopsy were 22.9 +/- 21.7, 38.8 +/- 26.6 and 50.0 +/- 26.3, respectively, in the SCSH subgroup vs. 16.5 +/- 21.5, 27.6 +/- 28 and 33.6 +/- 30.3, respectively, in the GIS subgroup (P = 0.0051, P = 0.0005 and P = 0.0003, respectively). The technical failure rate was 5% in the SCSH subgroup vs. 2% in the GIS subgroup (P = 0.1522). CONCLUSIONS: GIS and SCSH showed similar technical feasibility. The procedure-related pain reported by patients during contrast sonohysterography, as well as during subsequent hysteroscopy and endometrial sampling, was less in the GIS group. PMID- 19902476 TI - Does the size of three-dimensional power Doppler spherical sampling affect the interobserver reproducibility of measurements of vascular indices in adnexal masses? PMID- 19902477 TI - Cellular uptake and toxicity of gold nanoparticles in prostate cancer cells: a comparative study of rods and spheres. AB - Using a series of gold nanoparticles with incremental increase in dimensions but varying geometries (spherical vs rods) we have evaluated the influence of shape, size, surface properties and concentration on cellular uptake, adsorption of proteins and toxicity in a human prostate cancer cell line (PC-3). In the range of 30-90 nm diameter studied, spherical particles of 50 nm in diameter without polyethylene glycol (PEG) had the highest uptake. Surface attachment of PEG reduced cellular uptake. PEGylated gold nanorods had a net positive charge compared with their spherical counterparts and particle geometry influenced cellular uptake. In the absence of serum proteins the uptake of plain spherical GNPs increased. These studies pave the way for the tailoring of gold nanoparticles for targeted tumor therapy applications. PMID- 19902478 TI - Rearrangement and predissociation processes in negative molecular ions of nitrobenzenes. AB - The reactions of resonant electron capture by the molecules of benzene nitroderivatives has been studied in the gas phase. Some fragment negative ions were found to be unstable with respect to electron autodetachment. This circumstance has been used for the determination of their structure. In particular, it has been established that the low measured appearance energy of neutral component of [M-H](-) ion beam is a result of isomerization of nitrobenzenes' molecular ion, leading to the 2-nitrosophenol structure with the subsequent formation of the phenoxide anion in the autodetaching state. The effective yield curves of some types of fragment ions demonstrate fine vibrational structures, testifying the predissociation mechanism of ion formation. For all detected ions, the absolute cross sections of formation have been measured. PMID- 19902479 TI - Robot-assisted staging using three robotic arms for endometrial cancer: comparison to laparoscopy and laparotomy at a single institution. AB - PURPOSE: To demonstrate the feasibility of robot-assisted staging surgery using three arms in patients with endometrial cancer. METHODS: One hundred nine patients with clinical stage I endometrial cancer who underwent staging surgery at Yonsei University Health System were enrolled from May 2006 to January 2009. Patient demographics and operative outcomes were prospectively collected. RESULTS: Robotic surgery using three arms was performed in 28 patients, laparoscopy in 25, and laparotomy in 56. There were no differences among the three groups in terms of patient demographics. The number of harvested pelvic lymph nodes was lower in the laparoscopy group than in the laparotomy group (18.36 +/- 7.25 vs. 24.39 +/- 10.08, respectively, P = 0.025), but there was no difference between the robot and laparotomy groups. The number of resected para aortic lymph nodes and operative time did not differ among the three groups. The average hospital stay was longer for the laparotomy group than the robot and laparoscopy groups (10.78 days vs. 7.92 days vs. 7.67 days, respectively, P < 0.001). Operative complications and transfusions developed more frequently in the laparotomy group than in the robot and laparoscopy groups (25.0% vs. 7.1% vs. 8.0%, respectively, P = 0.049; 42.9% vs. 14.3% vs. 16.0%, respectively, P = 0.006). CONCLUSION: Robot-assisted surgery using three arms is a feasible method for surgical staging in patients with clinical stage I endometrial cancer. PMID- 19902480 TI - Toxicant-associated steatohepatitis in vinyl chloride workers. AB - Although nonalcoholic steatohepatitis (NASH) is typically associated with obesity, it has also been reported to occur in lean individuals exposed to industrial chemicals. Occupational exposure to vinyl chloride (VC) is a well documented risk factor for hemangiosarcoma, but has not previously been associated with steatohepatitis. Here we evaluate liver biopsies from 25 nonobese, highly exposed VC workers for steatohepatitis. Next, we evaluate associated metabolic and cytokine abnormalities in affected workers controlled by 26 chemical workers with no to minimal VC exposures, and 11 unexposed, healthy volunteers. Among highly exposed VC workers the prevalence of steatohepatitis was 80%. Of these, 55% had fibrosis and four had hemangiosarcoma. We have coined the term toxicant-associated steatohepatitis (TASH) to describe this condition, which was not explained by obesity or alcohol. Although mean serum transaminases were normal in TASH, total cytokeratin 18, but not the caspase-cleaved fragment, was elevated. Despite the absence of obesity, workers with TASH had insulin resistance with reduced adiponectin levels. TASH was also associated with markedly elevated serum tumor necrosis factor alpha and interleukins 1beta, 6, and 8. Serum antioxidant activity was reduced in TASH. CONCLUSION: TASH occurred frequently in these nonobese VC workers with high cumulative exposures and normal liver enzymes. Elevated total cytokeratin 18 suggested the presence of necrotic cell death in TASH and may be a useful serologic biomarker. TASH was further characterized by insulin resistance, elevated proinflammatory cytokines, and impaired antioxidant defenses. The threshold VC exposure and the role of other chemical agents in TASH are as yet unknown. PMID- 19902481 TI - Toll-like receptor 9 inhibition confers protection from liver ischemia reperfusion injury. AB - Endogenous ligands such as high-mobility group box 1 (HMGB1) and nucleic acids are released by dying cells and bind Toll-like receptors (TLRs). Because TLR9 sits at the interface of microbial and sterile inflammation by detecting both bacterial and endogenous DNA, we investigated its role in a model of segmental liver ischemia-reperfusion (I/R) injury. Mice were subjected to 1 hour of ischemia and 12 hours of reperfusion before assessment of liver injury, cytokines, and reactive oxygen species (ROS). Wild-type (WT) mice treated with an inhibitory cytosine-guanosine dinucleotide (iCpG) sequence and TLR9(-/-) mice had markedly reduced serum alanine aminotransferase (ALT) and inflammatory cytokines after liver I/R. Liver damage was mediated by bone marrow-derived cells because WT mice transplanted with TLR9(-/-) bone marrow were protected from hepatic I/R injury. Injury in WT mice partly depended on TLR9 signaling in neutrophils, which enhanced production of ROS, interleukin-6 (IL-6), and tumor necrosis factor (TNF). In vitro, DNA released from necrotic hepatocytes increased liver nonparenchymal cell (NPC) and neutrophil cytokine secretion through a TLR9 dependent mechanism. Inhibition of both TLR9 and HMGB1 caused maximal inflammatory cytokine suppression in neutrophil cultures and conferred even greater protection from I/R injury in vivo. CONCLUSION: TLR9 serves as an endogenous sensor of tissue necrosis that exacerbates the innate immune response during liver I/R. Combined blockade of TLR9 and HMGB1 represents a clinically relevant, novel approach to limiting I/R injury. PMID- 19902482 TI - Prevalence of hepatitis B surface antigen among refugees entering the United States between 2006 and 2008. AB - The Centers for Disease Control and Prevention recommends hepatitis B surface antigen (HBsAg) testing to identify chronic hepatitis B virus infection for foreign-born persons from countries or regions with HBsAg prevalence of >or=2%. However, limited data exist to indicate which countries meet this definition. To address this data gap, we estimated the HBsAg prevalence among refugees entering the United States between 2006 and 2008. We contacted state refugee health coordinators and asked them to report the number of refugees, country of origin, and HBsAg prevalence among refugees screened in their jurisdiction during the most recently available 12-month period prior to August 2008. We pooled data across jurisdictions and calculated the prevalence for any country with more than 30 refugees entering the United States, and where this level of data was not available by country, continents were considered. Of the 47 jurisdictions contacted, we received basic information from 31, with nine jurisdictions reporting HBsAg prevalence by country of origin applicable to 31,980 refugees (approximately 42% of refugees entering the United States during the observation period). We estimated an HBsAg prevalence of 2.8% (95% confidence interval 2.6% 3.0%) for refugees overall. Of the 37 countries with 30 or more refugees entering the United States, 25 had a prevalence of >or=2%. Prevalence was highest among refugees from Africa and Southeast Asia, and lowest among refugees from the Middle East and South/Central America. In the eight countries for which we had comparison data, six had lower HBsAg prevalence than in 1991. PMID- 19902483 TI - Activated monocytes in peritumoral stroma of hepatocellular carcinoma promote expansion of memory T helper 17 cells. AB - Although cancer patients exhibit a generalized immunosuppressive status, substantial evidence indicates that the inflammatory reaction at a tumor site can promote tumor growth and progression. Hepatocellular carcinoma (HCC) is usually derived from inflamed cirrhotic liver with extensive leukocyte infiltration. We recently found that proinflammatory T helper (Th)17 cells are accumulated in HCC tissue, where they promote disease progression by fostering angiogenesis. Here we show that interleukin (IL)-17-producing cells were enriched predominantly in peritumoral stroma of HCC tissues, and their levels were well correlated with monocyte/macrophage density in the same area. Most peritumoral CD68(+) cells exhibited an activated phenotype. Accordingly, tumor-activated monocytes were significantly superior to the suppressive tumor macrophages in inducing expansion of Th17 cells from circulating memory T cells in vitro with phenotypic features similar to those isolated from HCCs. Moreover, we found that tumor-activated monocytes secreted a set of key proinflammatory cytokines that triggered proliferation of functional Th17 cells. Inhibition of monocytes/macrophages inflammation in liver markedly reduced the level of tumor-infiltrating Th17 cells and tumor growth in vivo. CONCLUSION: The proinflammatory Th17 cells are generated and regulated by a fine-tuned collaborative action between different types of immune cells in distinct HCC microenvironments, and allows the inflammatory response of activated monocytes to be rerouted in a tumor-promoting direction. Selectively modulating the "context" of inflammatory response in tumors might provide a novel strategy for anticancer therapy. PMID- 19902484 TI - The Role of Transjugular Intrahepatic Portosystemic Shunt (TIPS) in the Management of Portal Hypertension: update 2009. PMID- 19902485 TI - Mouse organic solute transporter alpha deficiency enhances renal excretion of bile acids and attenuates cholestasis. AB - Organic solute transporter alpha-beta (Ostalpha-Ostbeta) is a heteromeric bile acid and sterol transporter that facilitates the enterohepatic and renal-hepatic circulation of bile acids. Hepatic expression of this basolateral membrane protein is increased in cholestasis, presumably to facilitate removal of toxic bile acids from the liver. In this study, we show that the cholestatic phenotype induced by common bile duct ligation (BDL) is reduced in mice genetically deficient in Ostalpha. Although Ostalpha(-/-) mice have a smaller bile acid pool size, which could explain lower serum and hepatic levels of bile acids after BDL, gallbladder bilirubin and urinary bile acid concentrations were significantly greater in Ostalpha(-/-) BDL mice, suggesting additional alternative adaptive responses. Livers of Ostalpha(-/-) mice had higher messenger RNA levels of constitutive androstane receptor (Car) than wild-type BDL mice and increased expression of Phase I enzymes (Cyp7a1, Cyp2b10, Cyp3a11), Phase II enzymes (Sult2a1, Ugt1a1), and Phase III transporters (Mrp2, Mrp3). Following BDL, the bile acid pool size increased in Ostalpha(-/-) mice and protein levels for the hepatic basolateral membrane export transporters, multidrug resistance-associated protein 3 (Mrp3) and Mrp4, and for the apical bilirubin transporter, Mrp2, were all increased. In the kidney of Ostalpha(-/-) mice after BDL, the apical bile acid uptake transporter Asbt is further reduced, whereas the apical export transporters Mrp2 and Mrp4 are increased, resulting in a significant increase in urinary bile acid excretion. CONCLUSION: These findings indicate that loss of Ostalpha provides protection from liver injury in obstructive cholestasis through adaptive responses in both the kidney and liver that enhance clearance of bile acids into urine and through detoxification pathways most likely mediated by the nuclear receptor Car. PMID- 19902487 TI - Bronchopulmonary fistula closure with an Amplatzer Multi-Fenestrated Septal Occluder. AB - Bronchopulmonary fistula, a communication between the bronchial airway and the pleural space, is associated with increased morbidity and mortality often requiring surgical therapy. A successful closure of a fistula from the posterior trachea to the right apical pleural space in a 60-year-old man with a history of Barrett's esophagus, esophagectomy, multiple pulmonary infections, and right upper lobectomy using an Amplatzer Multi-Fenestrated Septal Occluder via a transbronchial approach is reported. PMID- 19902486 TI - Mutation of bacterium Vibrio gazogenes for selective preparation of colorants. AB - A novel marine bacterium strain effectively produced prodiginine type pigments. These colorants could dye wool, silk and synthetic fabrics such as polyester and polyacrylic and also show antibacterial properties against Escherichia coli and Staphylococcus aureus bacteria on the dyed products. Methyl nitrosoguanidine was used as a mutation agent to increase the genetic diversity and the production yield of the bacteria of the family of Vibrio gazogenes. The analysis of the mutated samples showed that two new main colorants as well as three previously found ones were produced. Liquid chromatography electro spray ionization mass spectrometry (LC-ESI-MS) and nuclear magnetic resonance (NMR) spectroscopic techniques were used to elucidate the structures of the newly produced colorants. Mass measurements revealed that the colorants C1, C2, C3, C4 have molecular masses of 321, 323, 351, and 295 Da. One unstable colorant C5 with molecular mass of 309 Da was detected as well. The mutated bacteria strains increased the yield of pigment production by about 81% and produced prodigiosin in 97% purity. The antibiotic activities of pure colorants are discussed as well. Based on their bio activity and excellent dyeing capabilities, these colorants could be employed in cosmetic and textile industries. PMID- 19902488 TI - Cognitive and psychodynamic mechanisms of change in treated and untreated depression. AB - Two patient-level mechanisms of change, defenses, and cognition were tested over 3 time points in 65 depressed adults, approximately half receiving treatment. Early changes in automatic thoughts and immature defenses were associated with symptom change from time-one to time-three. The directionality of early automatic thought change predicting symptom change was partially supported, but immature defense change occurs simultaneously with, or after, symptom change. Given the convergent evidence of cognitive change as a mediator of depression reduction, all depression therapies should consider how they address depressive cognition. To build a more complete understanding of how to ameliorate depression, future studies should continue to include constructs from multiple theories and have measures of therapy process in addition to patient level mechanisms. PMID- 19902489 TI - Provision of social support to individuals with chronic fatigue syndrome. AB - The present study evaluated a buddy program designed to provide support for individuals with chronic fatigue syndrome (CFS). The intervention involved weekly visits by a student paraprofessional, who helped out with tasks that needed to be done in an effort to reduce some of the taxing demands and responsibilities that participants regularly encountered. This model of rehabilitation focused on avoiding overexertion in persons with CFS, aiming to avoid setbacks and relapses while increasing their tolerance for activity. Participants with CFS were randomly assigned to either a 4-month buddy intervention or a control condition. Posttest results showed that individuals who received a student buddy intervention had significantly greater reductions in fatigue severity and increases in vitality than individuals in the control condition. There were no significant changes between groups for physical functioning and stress. Buddy interventions that help patients with CFS reduce overexertion and possibly remain within their energy envelopes can be thought of as representing a different paradigm than nonpharmacologic interventions that focus only on increasing levels of activity through graded exercise. PMID- 19902490 TI - A comparison of the VASP index between patients with hemodynamically complicated and uncomplicated acute myocardial infarction. AB - INTRODUCTION: Critically-ill patients with ST-segment elevation myocardial infarction (STEMI) often present with insufficient gastroduodenal motility, liver hypoperfusion, and higher levels of circulating catecholamines. All of these factors can lead to reduced efficacy of clopidogrel, which is only available as a p.o. medication. The aim of the study was to compare clopidogrel effectiveness in unstable STEMI patients on mechanical ventilation with stable STEMI patients. MATERIALS AND METHODS: Two groups of twenty patients with STEMI were enrolled. One group (unstable) consisted of 20 hemodynamically unstable patients on mechanical ventilation and catecholamine support. The other group (stable) consisted of 20 control patients (all patients with STEMI in Killip I class). All patients were treated by primary Percutaneous coronary intervention. Blood samples were drawn before (baseline), at 4h (4h+), 24h (1d+) and 2 days (2d+) after clopidogrel administration. Clopidogrel efficacy was assessed by measurement of vasodilator-stimulated phosphoprotein phosphorylation index. RESULTS: The decrease in the vasodilator-stimulated phosphoprotein (VASP) index was substantially less in unstable patients compared with stable ones (ANOVA, P < 0.001). In stable patients, the VASP index decreased significantly by 20% at 4h+ and by 34% at 1d+, and remained significantly decreased by 31% at 2d+. In unstable patients, the VASP decreased nonsignificantly by 8% at 4h+, and no further decrease of VASP was present (-7% at 1d+, -11% at 2d+). CONCLUSIONS: Laboratory clopidogrel efficacy is lower in patients with MI and severe hemodynamic instability, probably due to splanchnic and liver hypoperfusion and catecholamine use. PMID- 19902492 TI - Dental-statistical meetings. PMID- 19902491 TI - Long-term health outcome and mortality evaluation after invasive coronary treatment using drug eluting stents with or without the IVUS guidance. Randomized control trial. HOME DES IVUS. AB - OBJECTIVE: To assess the role of the intravascular ultrasound (IVUS) during implantation of drug-eluting stents (DES) on long-term outcome in patients with complex coronary artery disease and high clinical risk profile with special attention to the development of late stent thrombosis (LST). METHODS: Two hundred and ten patients were randomly assigned to receive DES either with (N = 105) or without (N = 105) the IVUS guidance. Dual antiplatelet treatment was administered for 6 months in all patients. At 18-month follow-up, the rates of major adverse cardiac events (MACEs) (death, myocardial infarction, and reintervention) were assessed in both groups with special attention to possible LST. Stent thrombosis was classified according to Academic Research Consortium (ARC). RESULTS: At the 18-month follow-up, there was no significant difference between both groups regarding MACE (11% vs. 12%; P = NS). Stent thrombosis has occurred in four patients (3.8%) in the group with and in 6 patients (5.7%; P = NS) in the group without the IVUS guidance. CONCLUSIONS: In our randomized trial we failed to demonstrate the superiority of the IVUS guidance during DES implantation over standard high-pressure postdilatation. However we confirmed worrisome results concerning DES thrombosis after discontinuation of dual antiplatelet-treatment with documented stent thrombosis related events in almost 5% of patients with 50% of mortality in this high-risk clinical scenario. PMID- 19902493 TI - Adapting Haavikko's dental age for the assessment of Italian children: use of LMS and other models based on smoothing splines. AB - The expression 'dental age' (DA) denotes the mean age at which a given stage of dental development is achieved in a population chosen as calibrator. The rhythm at which the maturation occurs is a characteristic of a given population in a given period. When DA is determined on subjects of a population different from the calibrator, the shape of the relationship of DA to chronological age (CA) must be empirically determined to trace reference charts for DA that properly apply to this population. The aim of this study is to show how the DA determined in a calibration set (Haavikko's DA) can be adapted to apply to another population (Italian children), by means of CA-dependent smooth functions of the location, scale and shape of DA distribution. A set of 492 good quality panoramic films taken on healthy Italian children aged 5.5-14.5 years was analysed. The techniques under comparison produced quite similar curves: a parsimonious (4 e.d.f.) and rather satisfactory smoothing spline was found for median, but not for scale and shape indices: so the meaning of SDS values derived from these adjusted reference charts may result doubtful. Among the software products considered here, LMS program was found to be particularly user friendly, since allows the user to visually control the effect of changing the number of e.d.f. on the pattern of DA reference charts. PMID- 19902494 TI - Modelling count data with excessive zeros: the need for class prediction in zero inflated models and the issue of data generation in choosing between zero inflated and generic mixture models for dental caries data. AB - Count data may possess an 'excess' of zeros relative to standard distributions. Zero-inflated Poisson (ZiP) or binomial (ZiB) and generic mixture models have been proposed to deal with such data. We consider biomedical count data with an excess number of zeros and seek to address the following: (i) do zero-inflated models need covariates in the distribution part to predict class membership; (ii) what model-fit criteria have clinical relevance to predicted counts; (iii) can very different model parameterizations have near-identical fit; and (iv) how could model selection and hence model interpretation be aided by considering data generation processes? We show that covariates in the distribution part of zero inflated models are needed to predict class membership. A range of model-fit criteria should be considered, as consensus is rarely achieved, and considering predicted outcomes may be just as valuable as likelihood-based criteria. Zero inflated and generic mixture models may be indistinguishable according to both likelihood-based model-fit criteria and predicted outcomes, in which case model differentiation, hence, model selection and interpretation, might be guided by the consideration of a priori data generation processes. Zero-inflated models reflect whether or not there are (or have been) risk differences in disease onset and disease progression, while generic mixture models identify sub-types of individuals with similar risks of disease onset and progression. One or both modelling strategies may be used, though a priori knowledge or clinical impression of data generation might help to distinguish between two or more parameterizations that exhibit similar fit and yield near-identical predicted counts. PMID- 19902495 TI - An investigation of the MC-SIMEX method with application to measurement error in periodontal outcomes. AB - Measurement error is pervasive in medical research. In periodontal research studies, one measure of disease status is the probed pocket depth (PPD), the depth of the space between a tooth and the surrounding gum. In larger studies, these assessments are made by multiple examiners, each having distinct measurement error characteristics. Because PPD is recorded in whole millimeters, it may be regarded as discrete and its associated error as misclassification error. This study investigates the impact of this measurement error when evaluating the effect of periodontal disease status on levels of inflammatory markers in gingival crevicular fluid (GCF). The marker readings are either left or right censored, due to quantities that are either too small to be reliably quantified or so large that they saturate the detector. Additionally, marker readings from multiple periodontal sites within a subject's mouth are correlated. These considerations give rise to a clustered survival model for the marker readings in which the discrete predictor of interest is misclassified. Associations between the GCF markers and periodontal assessments are corrected for misclassification error using the MC-SIMEX method. Simulation studies reveal the impact of varying degrees of misclassification error on associations of interest. Analysis of pilot data from a periodontal study, for which examiner misclassification rates are estimated from calibration studies, further illustrates the approach. PMID- 19902496 TI - Survival analysis with clustered observations of orthodontic brackets. AB - Failures in the bonding of dental bracket are a big concern for orthodontists. Clinical experience suggests that some patients are more prone than others to experience failures. Therefore, it can be expected that in statistical analysis of orthodontic bracket failures, the usual assumption of independence between the observations is violated. An approach to overcome this problem is to apply the frailty model, in which the association between failure times is modelled with a random-effect term (i.e. frailty). We postulated that brackets of the same subject share the same frailty, that is, a latent common group effect, due to some unknown or unobserved covariates.The aim of this study was to investigate possible risk factors related to bracket failure using Cox proportional hazards model with a shared frailty term and to compare the results with those obtained using a basic Cox proportional hazards model.Survival data for 1677 brackets were obtained from a cohort of 54 females (mean age +/- SD: 13.3+/-4.8 yrs) and 46 males (mean age +/-SD: 13.1+/-3.8 yrs) over a eight-year period. Age, gender, vertical craniofacial morphology and anatomical location of brackets were entered into Cox models as covariates. The findings indicated that bracket failure was significantly affected by tooth position within the dental arch, with the highest failure risk in maxillary posterior region. Age, gender and vertical craniofacial morphology did not affect bracket failure. A Cox proportional hazards model with a shared frailty term represents a useful approach for modelling orthodontic bracket failures. PMID- 19902497 TI - Flexibility of Bayesian generalized linear mixed models for oral health research. AB - Many outcome variables in oral research are characterized by positive values and heavy skewness in the right tail. Examples are provided by many distributions of dental variables such as DMF (decayed, missing, filled teeth) scores, oral health impact profile score, gingival index scores, and microbiologic counts. Moreover, heterogeneity in data arises when more than one tooth is studied for each patient, due to the clusterization.Over the past decade, linear mixed models (LMEs) have become a common statistical tool to account for within-subject correlation in data with repeated measures. When a normal error is reasonably assumed, estimates of LMEs are supported by many statistical packages. Such is not the case for skewed data, where generalized linear mixed models (GLMMs) are required. However, the current software available supports only special cases of GLMMs or relies on crude Laplace-type approximation of integrals. In this study, a Bayesian approach is taken to estimate GLMMs for clustered skewed dental data. A Gamma GLMM and a log-normal model are employed to allow for heterogeneity across clusters, deriving from the patient-operator-tooth susceptibility typical of this clinical context. A comparison to the frequentist framework is also provided. In our case, Gamma GLMM fits data better than the log-normal distribution, while providing more precise estimates compared with the likelihood approach. A key advantage of the Bayesian framework is its ability to readily provide a flexible approach for implementation while simultaneously providing a formal procedure for solving inference problems. PMID- 19902500 TI - Size matters; even with sheathless guide catheters. PMID- 19902498 TI - Bayesian modeling of multivariate spatial binary data with applications to dental caries. AB - Dental research gives rise to data with potentially complex correlation structure. Assessments of dental caries yield a binary outcome indicating the presence or absence of caries experience for each surface of each tooth in a subject's mouth. In addition to this nesting, caries outcome exhibit spatial structure among neighboring teeth. We develop a Bayesian multivariate model for spatial binary data using random effects autologistic regression that controls for the correlation within tooth surfaces and spatial correlation among neighboring teeth. Using a sample from a clinical study conducted at the Medical University of South Carolina, we compare this autologistic model with covariates to alternative models to demonstrate the improvement in predictions and also to assess the effects of covariates on caries experience. PMID- 19902501 TI - The conserved active site tryptophan of thioredoxin has no effect on its redox properties. AB - In Staphylococcus aureus thioredoxin (Trx) it has been shown that mutation of the conserved active site tryptophan residue (Trp28) has a large effect on the protein stability, on the pKa of the nucleophilic cysteine and on the redox potential. Since these effects can either be due to the partially unfolding of the Trp28Ala mutant or to the absence of the indole side chain of Trp28 as possible interaction partner for the active site cysteines, the origin of the experimentally observed effects is not known and is beyond experimental approach. With theoretical pKa and density functional theory reactivity analysis on model systems where Trp28 has been replaced by an alanine within the structural environment of Trx it is shown that Trp28 does not affect the redox parameters of Trx. As such, the experimentally observed redox effects of the Trx W28A mutant might be due to structural changes induced by partial unfolding. PMID- 19902502 TI - Retrograde recanalization--RWT technique. PMID- 19902503 TI - Cardiac enzyme elevations after apparently successful percutaneous interventions are a marker of extensive coronary artery disease and complex stenoses. PMID- 19902504 TI - Quality metrics for each component of the STEMI care system. PMID- 19902505 TI - Small development but big contribution. PMID- 19902506 TI - Noncardiac surgery after coronary stenting or angioplasty [corrected] . PMID- 19902507 TI - High-resolution magnetic resonance angiography in the mouse using a nanoparticle blood-pool contrast agent. AB - High-resolution magnetic resonance angiography is already a useful tool for studying mouse models of human disease. Magnetic resonance angiography in the mouse is typically performed using time-of-flight contrast. In this work, a new long-circulating blood-pool contrast agent-a liposomal nanoparticle with surface conjugated gadolinium (SC-Gd liposomes)-was evaluated for use in mouse neurovascular magnetic resonance angiography. A total of 12 mice were imaged. Scan parameters were optimized for both time-of-flight and SC-Gd contrast. Compared to time-of-flight contrast, SC-Gd liposomes (0.08 mmol/kg) enabled improved small-vessel contrast-to-noise ratio, larger field of view, shorter scan time, and imaging of venous structures. For a limited field of view, time-of flight and SC-Gd were not significantly different; however, SC-Gd provided better contrast-to-noise ratio when the field of view encompassed the whole brain (P < 0.001) or the whole neurovascular axis (P < 0.001). SC-Gd allowed acquisition of high-resolution magnetic resonance angiography (52 x 52 x 100 micrometer(3) or 0.27 nL), with 123% higher (P < 0.001) contrast-to-noise ratio in comparable scan time ( approximately 45 min). Alternatively, SC-Gd liposomes could be used to acquire high-resolution magnetic resonance angiography (0.27 nL) with 32% higher contrast-to-noise ratio (P < 0.001) in 75% shorter scan time (12 min). PMID- 19902508 TI - An IVUS look for impossible chronic total occlusions. PMID- 19902509 TI - As goes the access, so goes the intervention? PMID- 19902510 TI - Patent foramen ovale closure and subsequent atrial fibrillation. PMID- 19902511 TI - Pediatric cardiovascular interventions: the good, the bad, and the ugly. PMID- 19902512 TI - Diastolic transmitral valve pressure gradients in patients with severe calcific aortic stenosis: more on the hemodynamics of the aging heart. PMID- 19902513 TI - Never say never, but tread lightly through vena cava filters. PMID- 19902514 TI - Motorboat or canoe? Impeller, propeller, or paddle? PMID- 19902515 TI - Online real-time reconstruction of adaptive TSENSE with commodity CPU/GPU hardware. AB - Adaptive temporal sensitivity encoding (TSENSE) has been suggested as a robust parallel imaging method suitable for MR guidance of interventional procedures. However, in practice, the reconstruction of adaptive TSENSE images obtained with large coil arrays leads to long reconstruction times and latencies and thus hampers its use for applications such as MR-guided thermotherapy or cardiovascular catheterization. Here, we demonstrate a real-time reconstruction pipeline for adaptive TSENSE with low image latencies and high frame rates on affordable commodity personal computer hardware. For typical image sizes used in interventional imaging (128 x 96, 16 channels, sensitivity encoding (SENSE) factor 2-4), the pipeline is able to reconstruct adaptive TSENSE images with image latencies below 90 ms at frame rates of up to 40 images/s, rendering the MR performance in practice limited by the constraints of the MR acquisition. Its performance is demonstrated by the online reconstruction of in vivo MR images for rapid temperature mapping of the kidney and for cardiac catheterization. PMID- 19902516 TI - Homozygous familial hypercholesterolemia: long term clinical course and plasma exchange therapy for two individual patients and review of the literature. AB - Familial hypercholesterolemia (FH) is an autosomal dominant disease. Homozygous FH (HFH) manifests with severe hypercholesterolemia since birth (cholesterol levels >5-6 the upper normal limit), which, if untreated, leads to early onset accelerated atherosclerosis and premature coronary death, usually before the 2nd or 3rd decades of life. Various invasive procedures (iliocecal bypass, porto caval shunt, liver transplant, and gene therapy) have been introduced for lowering low density lipoprotein (LDL) aiming at reducing atherosclerosis and improving survival of HFH patients. Of all the various methods, LDL apheresis has become the most attractive. Although its impressive effect on LDL-C reduction is well established, its long-term (of more than 10 year) effect on the atherosclerotic process and specifically cardiac end-points in HFH is hardly documented. We herewith report on the longest term lipophoresis so far reported in two HFH patients, each treated with plasma-exchange and LDL-apheresis for more than 20 years. The observations provide an opportunity to focus on various aspects regarding not only the procedure itself but also its effect on various clinical endpoints. By this description together with reviewing the literature, we discuss several issues, some of them are generalized while others are individualized, dealing with the approach of long term LDL apheresis in HFH. PMID- 19902517 TI - Genotoxicity of a freshwater cyanotoxin, cylindrospermopsin, in two human cell lines: Caco-2 and HepaRG. AB - Cylindrospermopsin (CYN), a cyanotoxin produced by certain freshwater cyanobacteria, causes human intoxications and animal mortalities. CYN is a potent inhibitor of protein- and glutathione-synthesis. Preliminary evidence for in vivo tumor initiation has been found in mice but the mechanism remains unclear. Several in vitro and in vivo studies demonstrate that CYN is genotoxic and requires metabolic activation. In the present study, the genotoxicity of CYN was assessed in human hepatocyte and enterocyte cell lines, which are models for CYN target organs. The cytokinesis-block micronucleus assay was conducted on liver derived HepaRG cells and colon-derived Caco-2 cells. Each cell-type was exposed to CYN in both the differentiated and the undifferentiated states, and both with and without the cytochrome P450 inhibitor, ketoconazole, to determine the involvement of metabolism in CYN genotoxicity. CYN increased the frequency of micronuclei in binucleated cells (MNBNC) in both Caco-2 and HepaRG cells. Moreover, ketoconazole reduced both the genotoxicity and cytotoxicity caused by CYN. Our results confirm the involvement of metabolic activation of CYN in mediating its toxicity and suggest that CYN is progenotoxic. PMID- 19902518 TI - Therapeutic options of caesarean scar pregnancy: case series and literature review. AB - We describe our experience with the treatment of 4 caesarean scar pregnancies and provide an overview of current literature. Four women diagnosed with a caesarean scar pregnancy in our hospital between 1996 and 2007 were treated with local or systemic methotrexate and had a steady decline of the serum beta-hCG level. The uterus was preserved in all women and 3 of them had an uneventful subsequent pregnancy and delivery. We suggest that transcervical needle aspiration of amniotic fluid followed by intra-amniotic injection of methotrexate should be the treatment of choice, followed by surgical treatment only if methotrexate fails. PMID- 19902519 TI - Bootstrap method-based estimation of the minimum sample number for obtaining pharmacokinetic parameters in preclinical experiments. AB - Empirically, 3-6 samples at each sampling time point have been used for most preclinical one-point sampling experiments without any theoretical justification. The purpose of the present study is to propose a practical approach to determine the minimum sample number (N(min)) based on Monte Carlo simulation and a bootstrap resampling. A computer program MOMENT(BS), in which a bootstrap resampling algorithm is used to estimate mean and standard deviations of pharmacokinetic parameters, such as area under the curve and mean residence time, was applied to estimate N(min). A new simulation program, MONTE1, was developed to generate simulated data for bootstrap resampling using the model parameters including inter- and/or intra-individual variations. Then, an index, S(2)CV calculated as the sum of the squared coefficient of variation is proposed to determine the N(min). The proposed approach was applied to the actual data in preclinical experiments, and the usefulness of the approach was suggested. An issue that one-point sampling data cannot separately assess inter- and intra individual variability is discussed. PMID- 19902520 TI - Biodegradable PLGA based nanoparticles for sustained regional lymphatic drug delivery. AB - The purpose of this work is to evaluate biodegradable drug carriers with defined size, hydrophobicity, and surface charge density for preferential lymphatic uptake and retention for sustained regional drug delivery. PLGA-PMA:PLA-PEG (PP) nanoparticles of defined size and relative hydrophobicity were prepared by nanoprecipitation method. These were compared with PS particles of similar sizes and higher hydrophobicity. PLGA-PMA:PLGA-COOH (PC) particles at 80:20, 50:50, and 20:80 ratios were prepared by nanoprecipitation for the charge study. Particle size and zeta potential were characterized by dynamic light scattering and laser doppler anemometry, respectively. Particles were administered in vivo to rats subcutaneously. Systemic and lymph node uptake was evaluated by marker recovery. Lymphatic uptake and node retention of PP nanoparticles was shown to be inversely related to size. Lymphatic uptake and node retention of PP particles, as compared to PS particles, was shown to be inversely related to hydrophobicity. Lastly, lymphatic uptake and node retention of PC nanoparticles were directly related to the anionic charge on the particles. In vivo lymphatic uptake and retention in a rat model indicates that the 50 nm PP particles are ideal for sustained regional delivery into the lymphatics for prevention/treatment of oligometastases. PMID- 19902521 TI - The role of interferons in the treatment of osteosarcoma. AB - Interferons, a group of cytokines with antiangiogenic, direct antitumour and immunostimulating properties, have shown significant activity against osteosarcoma in vitro and in xenograft models. They have also been used in osteosarcoma clinical trials as a single adjuvant to surgery, with an apparent increase in relapse-free survival. In the ongoing EURAMOS 1 clinical trial, interferon alpha-2b is evaluated as an adjuvant treatment in osteosarcoma. This article reviews the rationale for the use of interferon in cancer with special reference to the treatment of osteosarcoma, including all published data of clinical efficacy in this disease. PMID- 19902522 TI - Getting a "Head Start": neuropsychological functioning of young children treated for brain tumors with delayed or no cranial radiation therapy. PMID- 19902523 TI - Definitions of the phenotypic manifestations of sickle cell disease. AB - Sickle cell disease (SCD) is a pleiotropic genetic disorder of hemoglobin that has profound multiorgan effects. The low prevalence of SCD ( approximately 100,000/US) has limited progress in clinical, basic, and translational research. Lack of a large, readily accessible population for clinical studies has contributed to the absence of standard definitions and diagnostic criteria for the numerous complications of SCD and inadequate understanding of SCD pathophysiology. In 2005, the Comprehensive Sickle Cell Centers initiated a project to establish consensus definitions of the most frequently occurring complications. A group of clinicians and scientists with extensive expertise in research and treatment of SCD gathered to identify and categorize the most common complications. From this group, a formal writing team was formed that further reviewed the literature, sought specialist input, and produced definitions in a standard format. This article provides an overview of the process and describes 12 body system categories and the most prevalent or severe complications within these categories. A detailed Appendix provides standardized definitions for all complications identified within each system. This report proposes use of these definitions for studies of SCD complications, so future studies can be comparably robust and treatment efficacy measured. Use of these definitions will support greater accuracy in genotype-phenotype studies, thereby achieving a better understanding of SCD pathophysiology. This should nevertheless be viewed as a dynamic rather than final document; phenotype descriptions should be reevaluated and revised periodically to provide the most current standard definitions as etiologic factors are better understood, and new diagnostic options are developed. PMID- 19902524 TI - Clinical and molecular features in patients with atypical teratoid rhabdoid tumor or malignant rhabdoid tumor. AB - The SMARCB1 gene status in 50 patients with atypical teratoid rhabdoid tumor and/or malignant rhabdoid tumor recruited to a German registry was prospectively analyzed with FISH and PCR. Altogether we found 40 SMARCB1 mutations in 28 patients. Two patients were positive for SMARCB1 staining at immunochemistry. Germline mutations were identified in 10 of 41 patients with CNS disease, including three large heterozygous deletions, six truncating mutations and one donor splice site mutation. No missense mutation was identified. Analysis of first degree relatives did not detect any carriers. Mutations were distributed over the SMARCB1-gene without particular clustering. No germline mutation was found in nine patients without CNS disease. Patients with germline mutation had a lower median age at diagnosis in comparison to those without detectable germline mutation (5.5 vs. 13 months, P = 0.001), a higher rate of primary multicentric CNS disease (5/10 vs. 5/36) and synchronous or metachronous mixed CNS and extracranial disease (4/10 vs. 1/36). Two year overall survival was 0% in patients with germline mutation and 48% in those without detectable germline mutation (P < 0.001). Patients with germline mutation of SMARCB1 manifest at an early age and have a very high risk for progression which has to be considered with respect to the outcome of further treatment studies. PMID- 19902525 TI - Ultrasound diagnosis of dicephalic conjoined twins at 24 weeks of gestation. AB - We report the case of dicephalic conjoined twins discovered incidentally on a routine ultrasound at 24 weeks of gestation. There were 2 heads and a neck that fused with 1 thorax, but the spines continued all the way to the coccyx. The spines were connected medially by a fused rib, and laterally, there were ribs that went around the thorax in a more normal fashion. Antenatal ultrasound images are supplemented by postnatal photographs and x-rays. PMID- 19902526 TI - Influence of excipients in comilling on mitigating milling-induced amorphization or structural disorder of crystalline pharmaceutical actives. AB - The feasibility of using excipients to suppress the amorphization or structural disorder of crystalline salbutamol sulphate (SS) during milling was investigated. SS was subjected to ball-milling in the presence of alpha-lactose monohydrate (LAC), adipic acid (AA), magnesium stearate (MgSt), or polyvinyl pyrrolidone (PVP). X-ray powder diffraction, dynamic vapor sorption (DVS), high sensitivity differential scanning calorimetry (HSDSC) were used to analyze the crystallinity of the milled mixtures. Comilling with crystalline excipients, LAC, AA, and MgSt proved effective in reducing the amorphization of SS. LAC, AA, or MgSt acting as seed crystals to induce recrystallization of amorphous SS formed by milling. During comilling, both SS and LAC turned predominantly amorphous after 45 min but transformed back to a highly crystalline state after 60 min. Amorphous content was below the detection limits of DVS (0.5%) and HSDSC (5%). Comilled and physical mixtures of SS and ALM were stored under normal and elevated humidity conditions. This was found to prevent subsequent changes in crystallinity and morphology of comilled SS:LAC as compared to significant changes in milled SS and physical mixture. These results demonstrate a promising application of comilling with crystalline excipients in mitigating milling induced amorphization of pharmaceutical actives. PMID- 19902527 TI - The functional roles of poly(ethylene glycol)-lipid and lysolipid in the drug retention and release from lysolipid-containing thermosensitive liposomes in vitro and in vivo. AB - Triggered release of liposomal contents following tumor accumulation and mild local heating is pursued as a means of improving the therapeutic index of chemotherapeutic drugs. Lysolipid-containing thermosensitive liposomes (LTSLs) are composed of dipalmitoylphosphatidylcholine (DPPC), the lysolipid monostearoylphosphatidylcholine (MSPC), and poly(ethylene glycol)-conjugated distearoylphosphatidylethanolamine (DSPE-PEG(2000)). We investigated the roles of DSPE-PEG(2000) and lysolipid in the functional performance of the LTSL doxorubicin formulation. Varying PEG-lipid concentration (0-5 mol%) or bilayer orientation did not affect the release; however, lysolipid (0-10 mol%) had a concentration-dependent effect on drug release at 42 degrees C in vitro. Pharmacokinetics of various LTSL formulations were compared in mice with body temperature controlled at 37 degrees C. As expected, incorporation of the PEG lipid increased doxorubicin plasma half-life; however, PEG-lipid orientation (bilayer vs. external leaflet) did not significantly improve circulation lifetime or drug retention in LTSL. Approximately 70% of lysolipid was lost within 1 h postinjection of LTSL, which could be due to interactions with the large membrane pool of the biological milieu. Considering that the present LTSL-doxorubicin formulation exhibits significant therapeutic activity when used in conjunction with mild heating, our current study provided critical insights into how the physicochemical properties of LTSL can be tailored to achieve better therapeutic activity. PMID- 19902528 TI - Propagation of uncertainty in nasal spray in vitro performance models using Monte Carlo simulation: part I. Model prediction using Monte Carlo simulation. AB - Design of experiment (DOE) methodology can provide a complete evaluation of the influences of nasal spray activation and formulation properties on delivery performance which makes it a powerful tool for product design purposes. Product performance models are computed from complex expressions containing multiple factor terms and response terms. Uncertainty in the regression model can be propagated using Monte Carlo simulation. In this study, four input factors, actuation stroke length, actuation velocity, concentration of gelling agent, and concentration of surfactant were investigated for their influences on measured responses of spray pattern, plume width, droplet size distribution (DSD), and impaction force. Quadratic models were calculated and optimized using a Box Behnken experimental design to describe the relationship between factors and responses. Assuming that the models perfectly represent the relationship between input variables and the measured responses, the propagation of uncertainty in both input variables and response measurements on model prediction was performed using Monte Carlo simulations. The Monte Carlo simulations presented in this article illustrate the propagation of uncertainty in model predictions. The most influential input variable variances on the product performance variance were identified, which could help prioritize input variables in terms of importance during continuous improvement of nasal spray product design. This work extends recent Monte Carlo simulations of process models to the realm of product development models. PMID- 19902529 TI - Interactions between quercetin and warfarin for albumin binding: A new eye on food/drug interference. AB - The interaction between quercetin, a popular antioxidant flavonoid, and human serum albumin (HSA) is investigated and characterized by means of induced circular dichroism and saturation transfer difference NMR. These techiques demonstrate the reversible binding of quercetin to the carrier protein, which is responsible for its dissolution in aqueous medium. Competition experiments with two classical probes for HSA binding sites, namely Ibuprofen and Warfarin (a common anticoagulant coumarin), demonstrate that quercetin has a primary binding site located in the subdomain IIA, where coumarins are hosted. The affinity for this site is large and we found that quercetin may effectively displace warfarin from HSA. This may have relevant consequences in rationalizing the interferences of common dietary compounds and food supplements to anticoagulant treatments. PMID- 19902530 TI - Absolute structural elucidation of natural products--a focus on quantum mechanical calculations of solid-state CD spectra. AB - In this review article we examine state-of-the-art techniques for the structural elucidation of organic compounds isolated from natural sources. In particular, we focus on the determination of absolute configuration (AC), perhaps the most challenging but inevitable step in the whole process, especially when newly isolated compounds are screened for biological activity. Among the many methods employed for AC assignment that we review, special attention is paid to electronic circular dichroism (CD) and to the modern tools available for quantum mechanics CD predictions, including TDDFT. In this context, we stress that conformational flexibility often poses a limit to practical CD calculations of solution CD spectra. Many crystalline natural products suitable for X-ray analysis do not contain heavy atoms for a confidential AC assignment by resonant scattering. However, their CD spectra can be recorded in the solid state, for example with the KCl pellet technique, and analyzed possibly by nonempirical means to provide stereochemical information. In particular, solid-state CD spectra can be compared with those calculated with TDDFT or other high-level methods, using the X-ray geometry as input. The solid-state CD/TDDFT approach, described in detail, represents a quick and reliable tool for AC assignment of natural products. PMID- 19902531 TI - Brief communication: The distribution of perikymata on Qafzeh anterior teeth. AB - Recent studies have suggested that Neandertals and modern humans differ in the distribution of perikymata (enamel growth increments) over their permanent anterior tooth crowns. In modern humans, perikymata become increasingly more compact toward the cervix than they do in Neandertals. Previous studies have suggested that a more homogeneous distribution of perikymata, like that of Neandertals, characterizes the anterior teeth of Homo heidelbergensis and Homo erectus as well. Here, we investigated whether Qafzeh anterior teeth (N = 14) differ from those of modern southern Africans, northern Europeans, and Alaskans (N = 47-74 depending on tooth type) in the percentage of perikymata present in their cervical halves. Using the normally distributed modern human values for each tooth type, we calculated Z-scores for the 14 Qafzeh teeth. All but two of the 14 Qafzeh teeth had negative Z-scores, meaning that values equal to these would be found in the bottom 50% of the modern human samples. Seven of the 14 would be found in the lowest 5% of the modern human distribution. Qafzeh teeth therefore appear to differ from those of modern humans in the same direction that Neandertals do: with generally lower percentages of perikymata in their cervical regions. The similarity between them appears to represent the retention of a perikymata distribution pattern present in earlier members of the genus Homo, but not generally characteristic of modern humans from diverse regions of the world. PMID- 19902532 TI - Secular change in heights of indigenous adults from a Zapotec-speaking community in Oaxaca, southern Mexico. AB - Secular change in adult height of residents in a rural indigenous community in the Valley of Oaxaca was evaluated. Subjects were measured in 1971 (49 males, 26 females 19-70 years), 1978 (128 males, 124 females 19-82 years) and 2000 (155 males, 255 females 19-89 years). Heights were adjusted for estimated loss with age using two protocols; height at 21 years of age was also estimated. The effects of age and secular factors on measured and adjusted heights were evaluated through segmented linear regressions for three birth periods, <1930, 1930 through 1959 and >or=1960 which approximate significant periods in Mexican history. Secular increase in height occurred but estimated rates varied over time and between sexes. Males born before 1930 showed a secular increase in height but females did not. Adults of both sexes born 1930-1959 showed secular gains and estimated rates did not differ. The secular gain in height continued among those born 1960 and later and estimated rates were similar in both sexes. Estimated height at 21 years of age increased in males (not significant) but not in females born before 1930, showed little or no change in those born between 1930-1959, and increased (not significant) in those born 1960 and later. Combining observations on adults with those for youth in the community indicated several phases of secular change in height that varied with years of birth. PMID- 19902533 TI - Symmetry-restrained molecular dynamics simulations improve homology models of potassium channels. AB - Most crystallized homo-oligomeric ion channels are highly symmetric, which dramatically decreases conformational space and facilitates building homology models (HMs). However, in molecular dynamics (MD) simulations channels deviate from ideal symmetry and accumulate thermal defects, which complicate the refinement of HMs using MD. In this work we evaluate the ability of symmetry constrained MD simulations to improve HMs accuracy, using an approach conceptually similar to Critical Assessment of techniques for protein Structure Prediction (CASP) competition: build HMs of channels with known structure and evaluate the efficiency of proposed methods in improving HMs accuracy (measured as deviation from experimental structure). Results indicate that unrestrained MD does not improve the accuracy of HMs, instantaneous symmetrization improves accuracy but not stability of HMs during subsequent unrestrained MD, while gradually imposing symmetry constraints improves both accuracy (by 5-50%) and stability of HMs. Moreover, accuracy and stability are strongly correlated, making stability a reliable criterion in predicting the accuracy of new HMs. Proteins 2010. (c) 2009 Wiley-Liss, Inc. PMID- 19902534 TI - Arteriovenous malformation in the pretragal region: case report. AB - BACKGROUND: Arteriovenous malformation (AVM) in the extracranial region is uncommon. The most common sites are cheek, ear, nose, and forehead in order of frequency in the extracranial region. Although the external ear is the second most common site for the extracranial AVM, AVM in the pretragal region has not been reported until now. METHODS AND RESULTS: We reported a case of AVM in the pretragal region, which was successfully removed by complete excision after preoperative selective embolization in a 32-year-old man. Combined superselective and particle embolization is a useful method to decrease the blood flow in the pretragal AVM. CONCLUSION: We successfully treated the patient for the AVM in the pretragal area by complete excision within 72 hours after the preoperative combined superselective and particle embolization. Although our case is similar to auricular AVMs in point of symptoms and treatment, this is the first case reported at this unusual site. PMID- 19902535 TI - Transoral CO(2) laser treatment for T(is)-T(3) glottic cancer: the University of Brescia experience on 595 patients. AB - BACKGROUND: Transoral CO(2) laser surgery has been accepted as a valuable therapeutic option for glottic cancer. METHODS: This was a retrospective analysis of 595 patients. Five-year overall and disease-specific survivals, local control with laser, locoregional, regional control, and organ preservation rates were calculated. The impact of different variables was calculated by univariate analysis. RESULTS: Overall, disease-specific and disease-free survivals, local control with laser, locoregional, regional control, and organ preservation rates were 87.5%, 99%, 81.3%, 92.7%, 98.9%, 98.2%, and 97.1%, respectively. Univariate analysis showed a significant impact of pT category on local control with laser, organ preservation, locoregional and regional control, of endoscopic re-treatment for positive deep surgical margins on local control with laser and organ preservation, and recurrence after endoscopic re-treatment on local control with laser and organ preservation. CONCLUSION: This series confirms the good oncologic outcomes of endoscopic laser surgery for T(is), T(1), and selected T(2) and T(3) glottic tumors. PMID- 19902536 TI - Expression of matrix metalloproteinase-1, -7, -9, -13, Ki-67, and HER-2 in epithelial-myoepithelial salivary gland cancer. AB - BACKGROUND: The expression of matrix metalloproteinases (MMPs) in epithelial myoepithelial salivary gland carcinoma has not been studied previously. METHODS: Immunohistochemistry for MMP-1, -7, -9, -13, Ki-67, and HER-2, as well as HER-2 gene amplification by silver enhanced in situ hybridization was performed in a series of 12 paraffin-embedded histopathologic samples of patients from Canada and Finland. RESULTS: A positive MMP-13 (p = .0022), higher MMP-13 (p = .0274), and higher MMP-9 (p = .0274) index (multiplication of staining intensity by percentage of the positive cells) predicted better overall survival. In disease specific analysis, higher MMP-9 index (p = .0327) predicted better survival. A higher volume corrected index (VCI) of Ki-67 (p = .0339) predicted worse disease specific survival. In 1 patient, HER-2 oncogene amplification was observed. CONCLUSION: MMPs and Ki-67 may have prognostic impact in epithelial-myoepithelial carcinoma. PMID- 19902538 TI - Lisa S. Krivackas. PMID- 19902539 TI - Single-port access laparoscopic surgery for complex Crohn's disease. PMID- 19902540 TI - Safety of influenza vaccination in patients with myasthenia gravis: a population based study. AB - Influenza vaccination has been associated with adverse events including Guillain Barre syndrome. Because the safety of influenza vaccination in patients with myasthenia gravis (MG) has not been established, some clinicians discourage vaccination for these patients. We explored whether the administration of influenza vaccine to patients with MG might increase the risk of myasthenic crisis. Using population-based healthcare data from Ontario, Canada, from 1992 to 2007, we utilized the self-matched, case-series method of detecting adverse events following vaccination. We studied patients with established myasthenia who were hospitalized for MG within 42 weeks of influenza vaccination. We defined the primary risk interval as the 6 weeks following vaccination. Between January 1, 1992 and March 31, 2006, we identified 3667 hospital admissions for MG. No seasonal trend in MG admissions was evident. In 513 instances, hospitalization occurred within 42 weeks following vaccination in patients previously diagnosed with MG. Among these patients, 266 (52%) were men, the median age was 74 years, and 86 (17%) had previously undergone thymectomy. The estimated relative incidence of admission for MG in the primary risk interval compared with the control interval was 0.84 (95% confidence interval 0.65-1.09). We found similar results in stratified analyses according to gender, age, and thymectomy status. Vaccination of patients with MG against influenza was not found to be associated with exacerbations of the disease. Our findings do not support the practice of withholding influenza vaccination in patients with MG. PMID- 19902541 TI - Outcome of medical treatment of stricturing and penetrating Crohn's disease: a retrospective study. AB - BACKGROUND: Outcomes of medical treatment in patients with stricturing and penetrating Crohn's disease (CD) are not well characterized. METHODS: Adults with stricturing and penetrating CD who underwent medical treatment from 2004 to 2008 were evaluated. We assessed response rates to medical treatment, time to relapse or surgery, and postoperative complications. RESULTS: In all, 53 patients underwent medical therapy. 60% had stricturing disease, 11% had penetrating, and 28% had both. Disease location was ileal in 38%, colonic in 2%, and ileocolonic in 60%. At 30, 60, and 90 days, 54%, 60%, and 64% experienced a response to medical therapy, respectively. At 30 days, 75% of patients with ileal CD responded to therapy compared to 38% of patients with ileocolonic CD (P = 0.026). Overall, 64% of patients required surgery. Patients with ileocolonic disease required surgery at 0.55 years versus 1.07 years in patients with ileal disease (P = 0.023). 24% of patients experienced an anastomotic leak, fistula, or abscess (IASC). 29% of patients with penetrating disease developed IASC compared to 6% of patients with stricturing disease (P = 0.047). 32% of patients on biologic therapy had IASC compared to 0% of those not on biologics (P = 0.059). CONCLUSIONS: The outcomes of medical treatment of stricturing or penetrating CD are poor, as 64% ultimately require surgery. Important factors that seem to be associated with either failed therapy include ileocolonic or colonic disease location. We report a high rate of IASC, especially in patients with penetrating disease and those treated with biologic therapy. This should be considered prior to attempted medical therapy. PMID- 19902542 TI - Adaptive change in electrically stimulated muscle: a framework for the design of clinical protocols. AB - Adult mammalian skeletal muscles have a remarkable capacity for adapting to increased use. Although this behavior is familiar from the changes brought about by endurance exercise, it is seen to a much greater extent in the response to long-term neuromuscular stimulation. The associated phenomena include a markedly increased resistance to fatigue, and this is the key to several clinical applications. However, a more rational basis is needed for designing regimes of stimulation that are conducive to an optimal outcome. In this review I examine relevant factors, such as the amount, frequency, and duty cycle of stimulation, the influence of force generation, and the animal model. From these considerations a framework emerges for the design of protocols that yield an overall functional profile appropriate to the application. Three contrasting examples illustrate the issues that need to be addressed clinically. PMID- 19902543 TI - Microvascular free flap reconstruction versus palatal obturation for maxillectomy defects. AB - BACKGROUND: Palatal obturators and microvascular free flaps are both used to treat patients with maxillectomy defects, however, the optimal technique remains controversial. METHODS: A retrospective analysis of 113 patients undergoing maxillectomy for cancer was performed. Seventy-three patients received an obturator and 40 patients were reconstructed with a free flap. RESULTS: Speech intelligibility and postoperative diet were comparable between the obturator and free flap groups, except in cases of extensive (>50%) palatal defects, where free flap reconstruction was superior in both aspects (p = .019 and p = .043, respectively). The average time for presenting with a local recurrence in advanced cancer involving the palate was comparable in both groups (p = .33). CONCLUSION: Moderate-sized maxillectomy defects involving the palate can be successfully treated with either an obturator or free flap reconstruction. Extensive defects have a better functional outcome with free flaps. Evidence does not suggest that free flap reconstructions delay diagnosis of local recurrences. PMID- 19902544 TI - David D. Kilmer. PMID- 19902545 TI - Angiopoietin-2 in experimental colitis. AB - BACKGROUND: The pathophysiology of inflammatory bowel disease (IBD) includes leukocyte infiltration, blood and lymphatic remodeling, weight loss and protein enteropathy. The roles of angiopoietin-2 (Ang-2) in initiating gut inflammation, leukocyte infiltration and angiogenesis are not well understood. METHODS: Disease activity index, histopathological scoring, myeloperoxidase assay, immunohistochemistry and sodium dodecyl sulphate- polyacrylamide gel electrophoretic methods were employed in the present study to address the roles of Ang-2 in experimental colitis. RESULTS: Several important differences were seen in the development of experimental IBD in Ang-2(-/-) mice. Although weight change and disease activity differ only slightly in WT and Ang-2(-/-) + DSS treated mice, leukocyte infiltration, inflammation and blood and lymphatic vessel density is significantly attenuated compared to WT + DSS mice. Gut capillary fragility and water export (stool blood and form) appear significantly earlier in Ang-2(-/-) + DSS mice vs. WT. Colon lengths were also significantly reduced in Ang-2(-/-) and gut histopathology was less severe in Ang-2(-/-) compared to WT + DSS. Lastly, the decrease in serum protein content in WT + DSS was less severe in Ang-2(-/-) + DSS, thus protein losing enteropathy (PLE) a feature of IBD is relieved by Ang-2(-/-). CONCLUSION: These data demonstrate that in DSS colitis, Ang-2 mediates inflammatory hemangiogenesis, lymphangiogenesis and neutrophil infiltration to reduce some, but not all clinical features of IBD. The implications for Ang-2 manipulation in the development of IBD and other inflammatory diseases and treatments involving Ang-2 are discussed. PMID- 19902546 TI - Corneal confocal microscopy: a novel noninvasive means to diagnose neuropathy in patients with Fabry disease. AB - Neuropathy is a cause of significant disability in patients with Fabry disease, yet its diagnosis is difficult. In this study we compared the novel noninvasive techniques of corneal confocal microscopy (CCM) to quantify small-fiber pathology, and non-contact corneal aesthesiometry (NCCA) to quantify loss of corneal sensation, with established tests of neuropathy in patients with Fabry disease. Ten heterozygous females with Fabry disease not on enzyme replacement therapy (ERT), 6 heterozygous females, 6 hemizygous males on ERT, and 14 age matched, healthy volunteers underwent detailed quantification of neuropathic symptoms, neurological deficits, neurophysiology, quantitative sensory testing (QST), NCCA, and CCM. All patients with Fabry disease had significant neuropathic symptoms and an elevated Mainz score. Peroneal nerve amplitude was reduced in all patients and vibration perception threshold was elevated in both male and female patients on ERT. Cold sensation (CS) threshold was significantly reduced in both male and female patients on ERT (P < 0.02), but warm sensation (WS) and heat induced pain (HIP) were only significantly increased in males on ERT (P < 0.01). However, corneal sensation assessed with NCCA was significantly reduced in female (P < 0.02) and male (P < 0.04) patients on ERT compared with control subjects. According to CCM, corneal nerve fiber and branch density was significantly reduced in female (P < 0.03) and male (P < 0.02) patients on ERT compared with control subjects. Furthermore, the severity of neuropathic symptoms and the neurological component of the Mainz Severity Score Index correlated significantly with QST and CCM. This study shows that CCM and NCCA provide a novel means to detect early nerve fiber damage and dysfunction, respectively, in patients with Fabry disease. PMID- 19902547 TI - Fine-scale geographic variations of inflammatory bowel disease in France: correlation with socioeconomic and house equipment variables. AB - BACKGROUND: In a previous study we found a north-south gradient for Crohn's disease (CD) incidence in France. The aim of the present study was to determine if socioeconomic factors may influence the geographic distribution of CD and ulcerative colitis (UC) in France. METHODS: Using the national health insurance databases, incidence rates of CD and UC were estimated for each of 341 metropolitan "job areas" in 2000-2002. Relationships between incidence rates and relevant contextual variables from the 1999 French census were tested for significance using a Poisson regression. Mapping of smoothed relative risks (sRR) for CD and UC at the scale of job areas, using a Bayesian approach and adjusting for significant contextual variables, was carried out in order to search for geographic variations. RESULTS: CD incidence rates were negatively related to the percentage of farmers and to the percentage of housing with bathroom and toilets and positively related to the unemployment rate and to the percentage of households below the poverty threshold. Mapping of sRR for CD showed a clear north-south gradient, which was slightly improved after including the percentage of farmers and the percentage of housing with toilets. In UC we found no significant correlation between either incidence and socioeconomic variables or incidence and house equipment variables, and there was no north-south gradient. However, there was a positive and significant correlation between CD and UC incidence. CONCLUSIONS: The present study shows that geographic risk factors of CD in France are northern latitude, nonrural areas, and areas with poor sanitary house equipment. Among these factors the most important is northern latitude. PMID- 19902548 TI - TLR9 mRNA expression is upregulated in patients with active ulcerative colitis. PMID- 19902549 TI - A novel high-performance liquid chromatography/mass spectrometry method for improved selective and sensitive measurement of methotrexate polyglutamation status in human red blood cells. AB - The folate antagonist methotrexate is commonly used in low dose for treatment of rheumatoid arthritis and juvenile idiopathic arthritis. Therapeutic effects are attributed to intracellular levels of various methotrexate polyglutamates. The present methodology, combining a simple preparation step with ion-pairing reversed-phase liquid chromatography and electrospray ionization mass spectrometry, is suitable for the measurement of methotrexate and its polyglutamates(2-7), in human red blood cells. Sample preparation consists of perchloric acid protein precipitation followed by solid-phase extraction. Baseline separation of all analytes was achieved within 10 min using a Phenomenex Synergy C18 column together with a gradient solvent program obtained from blending acetonitrile with pH 7.5, 5 mM aqueous dimethylhexylamine. Seven methotrexate polyglutamates were detected using multiple reaction monitoring, with the mass spectrometer operating in positive ion mode. Using 20 microL injection volumes, limits of detection were 2.5 nM for individual methotrexate polyglutamates, while large volume (100 microL) injections led to detection limits of 0.5 nM and linear calibration from 0.5 to 100 nM for individual analytes. Finally, the presented methodology was applied for the analysis of methotrexate and its polyglutamates in red blood cells obtained from patients being treated for juvenile idiopathic arthritis with methotrexate. Significantly, the methodology proved suitable for determination of long-chain methotrexate polyglutamates(5-7) and further, appears to be superior with respect to sensitivity, selectivity and speed as compared to all previously described approaches. PMID- 19902550 TI - Detection of pathogenic Verticillium spp. using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Verticillium spp. have been listed by the European and Mediterranean Plant Protection Organization (EPPO) and China as plant quarantine pests. Although attempts have been made to develop a simple routine laboratory assay to detect these organisms, none are routinely used. We describe for the first time a robust assay for reliable identification of Verticillium spp. using protein fingerprinting data obtained by matrix-assisted laser desorption/ionization time of-flight mass spectrometry(MALDI-TOF-MS). Several sample preparation methods and matrices were investigated to improve mass spectra for the routine identification of six species of Verticillium spp.(Verticillium dahiliae, V. alboatrum, V. fungicola, V. nigrescens, and V. lecanii) by MALDI-TOF-MS. Using the optimized experimental method, we constructed a protein fingerprint database for six species of Verticillium and established a analysis criteria of log(Score). This MALDI-TOF-MS protocol should prove useful as a rapid and reliable assay for distinguishing different Verticillium spp. PMID- 19902551 TI - Comprehensive analysis of short peptides in sera from patients with IgA nephropathy. AB - We analyzed serum short peptides comprehensively to know whether they were useful to characterize IgA nephropathy (IgAN). Serum samples from 26 patients with untreated IgAN and 25 healthy donors were tested. Short peptides with molecular weights of approximately 7 kDa, purified from the serum samples by magnetic-beads based weak cation exchange, were detected by mass spectrometry. Then the peptide peaks detected were subjected to the multivariate data analysis by SIMCA-P+ containing principal component analysis (PCA) and orthogonal partial-least squares-discriminate analysis (OPLS-DA). A total of 92 peptide peaks were detected in the tested serum samples. The OPLS-DA analysis revealed that the profile of all the peptide peak intensities discriminated the IgAN group and the healthy group completely with a high R2 value (0.919) and a high Q2 value (0.861). Further, the profile of only five peptide peaks was found to discriminate the two groups. By tandem mass spectrometry and database searching, three of the five peptides which increased in the IgAN group were identified as fragments of fibrinogen alpha chain, and the two peptides which increased in the healthy group were identified as fragments of complement C3f and kininogen-1 light chain. Taken together, the profile of the serum short peptides would be useful to discriminate IgAN and healthy conditions. Further, the five peptides may be candidate serum markers for IgAN and may be related to pathogenesis of IgA. PMID- 19902553 TI - Dedication to Dr. Christopher J. O'Brien. PMID- 19902556 TI - Flow cytometric measurements of TB-specific T cells comparing with QuantiFERON-TB gold. AB - BACKGROUND: Interferon-gamma (IFN-gamma) release assays and the detection of IFN gamma synthesis in the cytoplasm of activated CD4+ T cells by flow cytometry have recently been used for tuberculosis (TB) diagnosis. The aim of this study was to compare the performance of IFN-gamma assay between ELISA (QuantiFERON-TB Gold, QFT) and intracellular cytokine flow cytometry (ICCFC), and to investigate the significance of an optimal gating strategy in ICCFC. METHODS: The CD4+ T cell response to TB antigens was measured using the intracellular cytokine staining technique and four color FC (CD3, CD4, IFN-gamma, and tumor necrosis factor-alpha (TNF-alpha)) on whole blood samples. The results were compared with those of QFT. RESULTS: Regarding sensitivity in the TB group, patients in the QFT positive TB group (N = 22) were all ICCFC positive and 9 patients (64%) in the QFT negative TB group (N = 14) were ICCFC positive. In all test tubes (N = 72), sensitivity of "targeted" gating for TNF-alpha+ IFN-gamma+ CD4+ T cells was significantly higher than customary gating (72%, 54%, respectively, P = 0.001). CONCLUSIONS: The diagnostic sensitivity of ICCFC was further confirmed to be much higher than that of QFT. In the ICCFC analysis, TNF-alpha+ IFN-gamma+ CD4+ T cells should be sequentially gated through appropriately defined regions, minimizing interferents and reflecting characteristics of light scatter and marker expressions of CD4+ T cells activated by TB antigens. PMID- 19902557 TI - When monocytes and platelets compete: The effect of platelet count on the flow cytometric measurement of monocyte CD36. AB - BACKGROUND: Flow cytometric measurement of monocyte surface CD36 is relevant to several conditions including diabetes, cardiovascular disease, lipid disorders, platelet isoimmunization, and susceptibility to P falciparum malaria. CD36 is also strongly expressed on platelets where it is also known as platelet glycoprotein IV. METHODS: Whole blood samples, containing identical monocyte concentrations, were adjusted to contain platelets ranging from 20,000/uL to 600,000/uL, were stained with fluorescent-labeled anti-CD36, and analyzed by flow cytometry. RESULTS: CD36 median fluorescent intensity (MFI) observed on monocytes decreased as the platelet concentration in the sample increased with more than a 50% decline in monocyte MFI over the normal range of platelet values. The effect was not abolished by using larger volumes of monoclonal antibody and was observed with different clones of reagent anti-CD36. The findings were most consistent with competition by platelets for the CD36 reagent. Similar findings were observed with antibody to class I HLA. Under defined assay conditions, monocyte CD36 MFI declined with rising platelet concentration in a predictable fashion following an inverse linear relationship. CONCLUSIONS: Measurement of CD36 expression on monocytes by flow cytometry in whole blood samples is affected by the sample platelet count. When comparing the monocyte CD36 expression among different individuals, our approach can be used to adjust measured monocyte CD36 expression for the effect of the platelet concentration in the sample. Competition by platelets for monoclonal reagents may occur in other settings when whole blood assays are used and when the target antigen is strongly expressed on both platelets and leukocytes. PMID- 19902558 TI - "STOP"-ping harm: when is state intervention justified? PMID- 19902561 TI - The utility of routine D-dimer measurement in syncope. AB - AIM: To establish whether D-dimer is an independent predictor of 1-month serious outcome and all-cause death in syncope patients presenting to the emergency department (ED). METHODS: This was a prospective cohort study of all adults presenting with syncope to an ED in the UK. Plasma D-dimer concentrations were determined in citrated plasma obtained at presentation. The primary endpoint was the combination of serious outcome and all-cause death at 1 month. RESULTS: Of 237 patients enrolled, 205 patients had a suitable plasma sample and had complete follow-up. Seventeen patients had a serious outcome or death at 1 month including three patients with a pulmonary embolus and eight with a serious cardiovascular outcome. Nearly a half of all patients (n=94; 46%) had a plasma D-dimer concentration above the upper limit of normal, 10 of whom had a serious outcome or death at 1 month. Receiver-operator characteristic curve analysis showed no relationship between plasma D-dimer concentration and serious outcome or death at 1 month. An elevated plasma D-dimer concentration was found in all patients with a pulmonary embolus. CONCLUSION: Plasma D-dimer is frequently raised in patients presenting with syncope to the ED and consequently does not predict 1-month serious outcome or death. We conclude that there is no role for the routine measurement of D-dimer in the management of patients presenting to the ED with syncope. PMID- 19902562 TI - Purine biosynthetic pathway genes and methotrexate response in rheumatoid arthritis patients among north Indians. AB - OBJECTIVE: Inter-individual variations to methotrexate (MTX) response among rheumatoid arthritis (RA) patients have been attributed to clinical heterogeneity and genetic variations influencing MTX pharmacology. In this study, we analyzed the association of polymorphisms in ATIC, AMPD1, ADA, and ADORA2A from the purine biosynthetic pathway with MTX response in RA patients from north India. We also assessed the cumulative contribution of these polymorphisms together with those from the receptor-metabolizer-transporter and folate pathway genes that we have previously investigated. METHODS: RA patients recruited using the American College of Rheumatology criteria were grouped into good (n = 213) and poor (n = 68) responders to MTX, based on Disease Activity Score 28-3. Individual single nucleotide polymorphism association was tested using (chi)2 test, and cumulative contribution of all the single-nucleotide polymorphisms and cumulative contribution of all the SNPs and clinico-demographic factors were assessed using linear and logistic regression. RESULTS: G allele of ADA rs244076 [P = 0.02, odds ratio (95% confidence interval): OR (95% CI) = 1.66 (1.01-2.75)]; and T allele of ADORA2A rs5751876 [P = 0.04, OR (95% CI) = 1.55 (1.01-2.37)] were associated with poor response, but did not stand Bonferroni correction. On regression analyses, FPGS rs1544105, TYMS rs2853539, DHFR rs7387, and ADA rs244076 were identified as putative predictors for MTX response. Carriers of the FPGS rs1544105 AA and AG genotypes [OR (95% CI) = 3.47 (1.19-10.12)] and TYMS rs2853539 AA genotype [OR (95% CI) = 2.76 (1.50-5.07)] were predictors of poor response in our patient population. CONCLUSION: Genes from all the three pathways seem to contribute to MTX response in the Indian population. However, these observations need to be replicated in an independent sample set. PMID- 19902563 TI - Proceedings of the 3rd ENETS Consensus Conference on Guidelines for the Standard of Care in Neuroendocrine Tumors, Palma de Mallorca, Spain, 2007. PMID- 19902564 TI - Clinical conundrum. Consider a case of poor growth in a young kitten. PMID- 19902566 TI - Putting Europe on the scientific map. PMID- 19902565 TI - How to...Approach rabbit anaesthesia with confidence. PMID- 19902567 TI - Tailoring cancer therapy - validating basic science with the 'supertrial'. AB - A paradigm shift in research culture is needed in order to set up the large ambitious trials needed to make personalised medicine a reality. Emma Wilkinson talks to the experts about the hurdles they face and what needs to change. PMID- 19902568 TI - Proceedings of the 6th International Conference on Basic and Therapeutic Aspects of Botulinum and Tetanus Toxins, June 12-14, 2008, Baveno, Italy. PMID- 19902569 TI - Proceedings of Food Micro 2008 "Evolving Microbial Food Safety and Quality" 1-4 September 2008, Aberdeen, Scotland, UK. PMID- 19902570 TI - Show some understanding for fellow professionals. PMID- 19902571 TI - Grand Designs. PMID- 19902572 TI - Mental ill health and issues of stress. PMID- 19902573 TI - Grand designs. PMID- 19902574 TI - Dangerous liaisons. PMID- 19902575 TI - On the straight and narrow. PMID- 19902576 TI - Mind changers: the pseudo-patient study. PMID- 19902577 TI - Could attendance allowance be scrapped? PMID- 19902578 TI - New horizons opens up a new agenda for mental health but change won't happen overnight. PMID- 19902579 TI - Mental health practitioners' role in supporting people maintain their jobs. PMID- 19902580 TI - Viewpoint. New Horizons. PMID- 19902581 TI - It makes you think. PMID- 19902582 TI - Doctor's orders. PMID- 19902583 TI - Perspectives. Buying inappropriately to try to lift my depression. PMID- 19902584 TI - On the record. Robert Westhead. Interview by Alita Howe. PMID- 19902586 TI - Proceedings and abstracts of the 6th International Headache Seminary, Stresa, Italy, 29-30 May 2009. PMID- 19902585 TI - Tophaceous gout with congenital dyserythropoietic anemia. PMID- 19902587 TI - Syphilis is back. PMID- 19902588 TI - Cardiometabolic risk. PMID- 19902589 TI - A patients duty to follow up. PMID- 19902590 TI - Illegible documents and public hospitals. PMID- 19902591 TI - Microbial-host interaction: tolerance versus allergy. The 64th Nestle Nutrition Institute Workshop. Foreword. PMID- 19902592 TI - Apology legislation: the time has come. PMID- 19902593 TI - Building a strong profession. PMID- 19902594 TI - Surveillance spotlight: new "syndemic" paradigm for interprofessional management of chronic inflammatory disease. PMID- 19902595 TI - The valuation high ground. PMID- 19902596 TI - [Risk factors for suicide attempts in dual diagnosis patients]. AB - BACKGROUND: Patients suffering from both psychiatric disorders and substance abuse/addiction are titled "dual diagnosis patients" (DDP). Substance abuse/addiction is associated with suicidal behavior. Although our knowledge of substance abuse/addiction and suicide behavior is increasing, we lack sufficient knowledge of suicide among DDP. OBJECTIVES: (1) To compare the rate of suicide attempts among DDP and non-DDP; (2) To determine risk factors for suicide attempts in DDP. METHODS: Analysis of 3,433 consecutive admissions: men and women aged 18-65 years in our center (06/2003-06/2005). RESULTS: Of 848 DDPs' admissions, 197 (23.2%) were after suicide attempts, whereas 403 of 2558 non DDP's admissions (15.8%) were after suicide attempts (odds ratio [OR] = 1.6; 95% confidence interval [95% CI] = 1.3 - 1.9). The OR in the multiple analysis was 1.4 [95% CI] = 1.1 - 1.8). By multivariate regression analysis, the positive result for Tetrahydrocannabinol (THC) in the urine analysis was a protective factor and a diagnosis of disorders of adult personality and behavior (according to the International Classification of Disease - 10 edition [ICD-10]) was an independent risk factor for suicide attempts. CONCLUSIONS: DDP have greater risks of suicide attempts than non-DDP. A comprehensive plan of preventive interventions for multidisciplinary staff is recommended in order to reduce suicide rates in DDP. PMID- 19902597 TI - [Recurrent hospitalizations due to false positive troponin in a patient presenting with chest pain]. AB - A 58 year old male patient was hospitalized numerous times due to an elevated troponin I level in spite of the absence of specific chest pain and ischemic changes on ECG. The patient had undergone two coronary angiography procedures and numerous cardiac studies without evidence of ischemic heart disease. The elevated troponin level remained unchanged in between hospitalizations while the patient was asymptomatic. Further workup revealed that the troponin I level was normal when tested with an alternative laboratory kit. The troponin T level was also normal. This case report underscores the problematic nature of relying on an elevated troponin level as the sole component in the diagnosis of coronary disease. PMID- 19902598 TI - [Planned home deliveries in Israel between the years 2003-2007]. AB - OBJECTIVE: To determine obstetric and perinatal outcome of planned home deliveries in Israel. METHODS: A retrospective study was performed including planned home deliveries in Israel between the years 2003 and 2007. RESULTS: Data regarding 1749 planned home deliveries was retrieved. Of these, 1594 (91.1%) were managed successfully. The rate of cesarean deliveries was 3.3% and the rate of instrumental deliveries was 1.0%. No cases of maternal mortality were noted. However, one patient was hospitalized for more than 5 days due to cesarean complications. One case of sudden infant death syndrome occurred 30 hours after home delivery. CONCLUSION: With proper selection of low risk parturients, planned home deliveries are basically associated with favorable outcomes. Further prospective studies should substantiate our results in order to provide clear indications for home deliveries. PMID- 19902599 TI - [Posterior lumbar interbody fusion (PLIF stand-alone) for chronic low back pain]. AB - OBJECTIVE: To evaluate the fusion achieved by posterior lumbar interbody fusion- PLIF [B-TWIN, Disc-O-Tech) stand-alone in terms of intervertebral fusion on the basis of radiograph imaging. SUMMARY OF BACKGROUND DATA: Lumbar fusion is being used to reduce pain and decrease disability in patients with chronic low back pain. Different surgical techniques are available. The use of PLIF stand-alone has been described with controversial results. METHODS: From 2003 through 2006, 14 patients with chronic low back pain were operated on in our department. In order to evaluate the quality of fusion, flexion and extension, radiographs were performed. A difference of 4 degrees was considered as failure of fusion. RESULTS: A total of 14 patients, 5 male and 9 female mean age 56 years (43-72), underwent 15 lumbar interbody fusions. Fourteen lumbar interbody fusions (93%) had less than 4 degrees of difference between flexion and extension radiographs; the overall median score was 1.7 degrees, the male group had a median score of 2.9 and the female group 1.1 (P = 0.066]. All the patients with degenerative disc disease had scores above 1.7 degrees, 57% (4) of the patients with spondylolisthesis had scores above 1.7 degrees and 1 patient (16% of this group) with spinal stenosis had a score above 1.7 degrees (P = 0.096). CONCLUSIONS: A fusion rate of 93% was achieved. The findings suggested a tendency for better results in the female group and better results in the spinal stenosis group compared with the spondylolisthesis group and the degenerative disc disease group. There is a need for further investigation in order to establish our findings. PMID- 19902600 TI - [Identical, for better and for worse]. AB - We report on identical premature twins (monochorionic diamniotic). As fetuses, they both demonstrated sonographic and MRI evidence of cerebral bilateral ventriculomegaly. Neonatal brain US showed bilateral ventriculomegaly, similar in both twins. During follow-up, these physical and imaging similarities persisted. To the best of our knowledge, this is the first report on similar bilateral ventriculomegaly in Identical Twins (IT). A genetic origin of this finding in our IT is suggested. Should more cases of ventriculomegaly be reported in the future in several sets of IT, cerebral ventriculomegaly can then be added to the list of similar CNS features observed in identical twins. PMID- 19902601 TI - [Plagiarism in medical journals--an editor's responsibility]. PMID- 19902602 TI - [The burn array in Israel]. AB - Circa 2700 burn victims require hospitalization in Israel each year. Most of these burns happen as household accidents, caused by hot liquids and involve less than 20% of the total body surface area. Treatment is dictated by the burn severity. Patients with severe burns are referred to one of the five dedicated burn services complying with the ABA & EBA criteria, located at the tertiary trauma centers which are situated in accordance with Israel's geographic, demographic and security needs. In most centers, the joint treatment of the most severe cases, provided by plastic surgeons and intensive care specialists allows the use of advanced techniques and treatment protocols, improving outcome. In order to further improve the quality of care for the burn patients, to allow improved national capability to contend with scenarios of disaster and mass casualty, to provide a sound database for future planning and billing, and also to produce appropriate intervention plans, Israel should have a system similar to the US's National Burn Repository. PMID- 19902603 TI - [Burns--risk factors and treatment]. AB - Burn injuries are very frequent and afflict approximately 1% of the population yearly. They are a source of heavy medical burden to medical systems worldwide. In the US alone, about 2 million burns are treated by medical staff yearly, and about 75,000 burns are serious enough to require hospitalization. In the UK, a similar situation is depicted in the statistics--burns constitute 1% of the ER workload, and 0.014% of the hospitalization. Morbidity and mortality from burns is mainly dependent upon: total body surface area (TBSA) that is involved in the burn, the depth of the burn and it's anatomical location, the age of the subject, prior medical history and the severity of adjacent injuries (especially pulmonological injury). TBSA is calculated by age-adjusted tables. There are a number of ways to determine this parameter, the simplest of all is called "the rule of 1/9". Using this technique we divide the body into distinct areas, each equal to 1/9 of the TBSA. The treatment of burn injuries is considered one of the most difficult in the medical profession and some even compare it to the treatment of ICU patients. The primary treatment in burns always involves the removal of the patient from the source of the thermal injury, securing his airway (especially in patients suspected of inhalation injury) and an aggressive fluid resuscitation. Fluid resuscitation is mainly managed using the Parkland equation. The treatment of the burned skin is by one of two regimes--the conservative regime (frequent redressing of the burn site, hygiene and antimicrobial treatment) and the surgical regime (early intervention with debridement, skin implantations etc.). Several different studies have shown a decrease in the mortality rate of severe burn patients who have undergone an early surgical regime in comparison to conservative treatment. PMID- 19902604 TI - [The no-reflow phenomenon following percutaneous coronary intervention]. AB - The no-reflow phenomenon after successful coronary artery perfusion is caused by a myocardial perfusion defect due to impaired blood flow in the microvascular bed. This phenomenon is more prevalent among acute myocardial infarction patients who have additional cardiovascular risk factors, and its presence constitutes an independent negative prognostic sign. The impaired microvascular myocardial blood flow is caused by microemboli and by intracellular processes taking place during the reperfusion phase. In the last few years, many studies have been published regarding new diagnostic and treatment modalities based on a better understanding of the pathophysiological processes implicated. PMID- 19902605 TI - [The approach to chronic pain management in the elderly]. AB - Chronic pain is a common, yet frequently overlooked and undertreated complaint among older patients. It may result in an impairment of the function and quality of life of these patients. Chronic pain is a syndrome, which influences and is influenced by many potential factors (physiological psychological and social), all of which require treatment to achieve an optimal clinical outcome. Effective management of chronic pain in this population is attainable, if properly recognized and treated. Reasons for inadequate pain control by clinicians include lack of training, inadequate pain assessment and reluctance to prescribe opiates. The approach to pain assessment and management is often different and more complex for older patients than for younger ones. In older people, a host of factors can impede assessment and drug management, including multiple medical problems and many potential sources of pain, sensory impairment, depression, disability and impaired cognitive function. In addition, pharmacokinetics and pharmacodynamics unique to the geriatric population make this group vulnerable to adverse medication effects. By using appropriate pharmacological combined with non-pharmacological treatment, it is often possible to improve function ability to a greater extent than the severity of pain reduced. PMID- 19902606 TI - [The broken heart syndrome]. AB - Tako-tsubo syndrome is characterized by acute myocardial depression secondary to psychological stress. The syndrome is ten times more common in females compared to males and is manifested by chest pain and dyspnea. ECG changes are similar to those seen in acute myocardial infarct. However, in this syndrome, cardiac enzyme levels are normal and coronary blood flow is not compromised. The syndrome is caused by excess of catecholamines. The prognosis is relatively favorable with full recovery in most patients. The recurrence rate is between 3 to 8% and there is no advantage in favor of specific treatment. This report reviews the recent literature and presents a patient who suffered from the syndrome. PMID- 19902607 TI - [Are you being served? The inter-organizational status and job perception of those responsible for patient rights in general hospitals in Israel]. AB - BACKGROUND: Paragraph 25 of the Patient's Rights Law in Israel requires that every medical facility director in Israel appoint an Ombudsman who will be responsible for patients' rights, receive patient complaints and resolve them. The law aims to strengthen the patient's position vis-a-vis service providers. Therefore, it is desirable that the Ombudsman shall function independently without apprehension or bias. OBJECTIVES: Eleven years after the law was legislated, the authors researched the following: Were individuals responsible for patient rights appointed in all general hospitals? Who are the position holders? What issues do they deal with? What is the weight of their various responsibilities concerning patient rights, as defined by the law, relative to their other tasks? Do they benefit from organizational mechanisms that assure their independence within the service provider's organization? How do they perceive their job--as "Patient Representatives" (as defined by law), or as representatives of their hospitals? METHODS: Hence, the authors personally interviewed each of the 26 General Hospital Directors in Israel as well as the Ombudsmen in each of their facilities. RESULTS: In each of Israel's general hospitals, an Ombudsman responsible for patient rights was appointed. In the majority of cases (82.6%) the Ombudsman was also engaged in an additional managerial or staff position within the organization. As a result, the Ombudsmen are almost entirely dependent on hospital management. The necessary means, by which to fulfill their positions and responsibilities as defined by the law, such as instructing and guiding medical staff regarding the protection of patient rights, have yet to be put at their disposal. The majority of the Ombudsmen view themselves as management representatives. These perceptions do not agree with the spirit of the Patient's Rights Law which is meant to strengthen the patient's position vis-a-vis medical services providers. The authors found a correlation between these views and the fact that Ombudsmen simultaneously hold additional managerial positions and some see themselves as part of their hospitals senior organizational hierarchy. In addition, we found a correlation between their seniority within the organization and their identification with the organization. CONCLUSIONS: It is recommended that the independence of those responsible for patient rights be strengthened by adding specific stipulations to the law on this matter and that the necessary means needed to fulfill their responsibilities as legislated, be put at their disposal. The authors recommend promoting the independent status of Ombudsmen by not imposing upon them responsibilities other than those for patient rights. PMID- 19902608 TI - [Determining treatment at the end of life: who decides?]. PMID- 19902609 TI - [Somerset Maugham--a doctor in human bondage]. PMID- 19902610 TI - Factors influencing the selection of a hospital for colon cancer surgery in Japan: analysis of the effects of surgery volume, hospital functions, and geographic conditions. AB - OBJECTIVE: To identify factors influencing the hospital choice of patients who undergo colon cancer surgery. METHODS: We conducted a multivariate analysis using micro data from the Patient Survey and the Survey of Medical Institutions conducted by the Statistics and Information Department, Minister's Secretariat, Ministry of Health, Labour and Welfare of Japan, to identify patient and hospital characteristics which influence a patient's decision whether to undergo surgery at a hospital where many colon surgeries have been performed ("high-volume hospital"). RESULTS: Hospitalizations to high-volume hospitals were linked to factors such as referral from nonmedical facilities and the fact that the medical institution was a teaching hospital. Hospitalizations to high-volume hospitals decreased by about 8% for every 1-km increase in distance to the nearest high volume hospital. CONCLUSION: Geographic factors and hospital function had an impact on admission to high-volume hospitals that are expected to contribute to functionalization of medical institutions and improvement of medical standards. This suggests that geographic conditions should be considered in planning regional coordination systems for specialized medical care. PMID- 19902611 TI - Oral health status in relation to stimulated saliva buffering capacity among Japanese adults above or below 35 years of age. AB - OBJECTIVES: To evaluate dental caries and periodontal pocketing in relation to saliva buffering capacity among two groups of Japanese adults aged above 35 years old (A35) or 35 years old and below (B35). METHODS: After measuring the initial pH of saliva, samples were titrated with 0.1N HCl to evaluate the buffer capacity. Levels of mutans streptococci and Lactobacilli in saliva, DMF and periodontal pocket were also measured. RESULTS: Among B35 group, DMF of adults with High buffering capacity was significantly lower than those of adults with Medium and Low buffering capacity (p < 0.05). Chi-square test showed the distribution of subjects with high counts of mutans Streptococci or Lactobacilli into the three buffer capacities, which was not statistically different between two age groups (p > 0.05). There were statistical differences for the mean periodontal pocket depth on the upper right incisor of adults in A35 with High buffer capacity and those of adults with any buffer capacity in B35 group. Statistical differences were also found for the pocket depth on lower left incisor and upper right molars of adults in A35 with High buffer capacity and those of adults with low buffer capacity in B35 group (p < 0.01). CONCLUSION: High buffering capacity of saliva might be associated with better status of oral health among Japanese adults in both age groups. PMID- 19902612 TI - A business of busyness. PMID- 19902613 TI - Night shift newcomers. PMID- 19902614 TI - Reentry problems. PMID- 19902615 TI - Relic or relevant? PMID- 19902616 TI - Defining alignment. PMID- 19902617 TI - Stepping off the wheel. PMID- 19902618 TI - Musings on busyness. PMID- 19902619 TI - A year out of the loop. PMID- 19902620 TI - Stepping stones. PMID- 19902621 TI - Service learning in rural communities. Medical students teach children about the brain. AB - Incorporating service learning into a medical school curriculum can have significant benefits for both the students and the communities they serve. The University of Minnesota Medical School-Duluth Campus has integrated an established neuroscience literacy program into a community service requirement for second-year medical students. Since 2005, medical students taking part in the program have made presentations about the brain and how it works to more than 10,000 elementary school students throughout Minnesota and northwestern Wisconsin. This article describes this initiative and the potential benefits to both the medical and elementary school students. PMID- 19902622 TI - Principles to promote physician satisfaction and work-life balance. AB - Substantial evidence suggests that difficulty balancing their personal and professional life is a major contributor to physician distress. Limited evidence suggests that the mission and policies of health care organizations may relate to physician satisfaction. In this article, we describe principles to promote professional satisfaction and work-life integration developed by the Mayo Clinic department of medicine. These principles can be used to measure and align policies. It is hoped they will serve as a model that can be used by other health care organizations. PMID- 19902623 TI - Lunch date. PMID- 19902624 TI - [Reproducibility of the fetal nasal bone measurement by means of bidimensional and three-dimensional ecography during the second trimester of the pregnancy]. AB - AIMS: The utility of the valuation-measurement of the nasal bone in the prognosis of chromosomopaties during the second trimester of the pregnancy is demonstrated. OBJECTIVE: To evaluate the repeatability of nasal bone measurement during second trimester with bidimensional and three-dimensional sonography. MATERIAL AND METHOD: Nasal bone was measured in 50 single pregnancies. First observer carried out two measures of nasal bone with bidimensional sonography, and 1 measure with three-dimensional sonography. Second observer carried out just one measure with bidimensional sonography (2D) and just another one with three-dimensional sonography (3D). We studied the intraobserver variability with 2D sonography, and the interobserver variability with 2D and 3D sonography. RESULTS: 2D-sonography: nasal bone measurement showed excellent intraobserver correlation with a correlation coefficient of 0,87 (CI 95%: 0,78-0,93) and a little means difference of 0,18 (SD: 0,74). Indeed, there was a good interobserver correlation with a correlation coefficient of 0,92 (CI 95%: 0,85-0,95), and a means difference of 0,14 (SD: 0,56). 3D-sonography: there was a acceptable interobserver correlation with correlation coefficient of 0,70 (CI 95%: 0,52-0,82). CONCLUSION: Nasal bone measurement is highly reproducible by means of bidimensional sonography while using three-dimensional sonography, the results are just acceptable. PMID- 19902625 TI - [Demographic impact of cesarean section]. AB - BACKGROUND: in the vital statistics the cesarean section is a cause that makes reference to births and deaths. OBJECTIVE: to analyze the frequency of the cesarean section, the changes brought about in the rates of maternal, perinatal mortality and of natality, and their relation with reliable contraceptive methods. PATIENTS AND METHOD: retrospective study realized with the dices of patients of the Coordination of Reproductive Health of the Mexican Institute of the Social Insurance (IMSS) of the state of Aguascalientes (Mexico). The births were registered majors of 20 weeks of gestation, the maternal and perinatal deaths from 1990 to 2007 in the hospitals of the IMSS in Aguascalientes. The rate of cesarean calculated by means of the registry of the number of the same realized per year and they were divided between the total of vaginal and abdominal births, and the result multiplied percent. RESULTS: 201,563 obstetrical events were registered, of which 145,106 corresponded to vaginal childbirths and 56,457 abdominal ones. With these data a global rate of cesarean section of 28% was obtained, with 201,182 new born alive ones and 2,618 perinatal ones. The acceptance of the tubary bilateral occlusion maintained a constant increase during the period of study, like the intrauterine device. The acceptance from the bilateral occlusion was completely different tubary during the cesarean one, in comparison with the childbirth. CONCLUSION: the long term results show positive influence in the reduction of the rate of natality. PMID- 19902626 TI - [Frequency of metabolic syndrome in Mexican postmenopausal women and its relation with the hormonal therapy]. AB - INTRODUCTION: Metabolic syndrome is related with a significant increase in some chronic-degenerative diseases. In the same manner in the postmenopause exist several and metabolic changes. OBJECTIVE: To determine the frequency of metabolic syndrome in Mexican postmenopausal women that receive or not hormone therapy. MATERIAL AND METHODS: 33 postmenopausal women divided in two groups one without hormone therapy and the other received conjugated equine estrogens plus medroxyprogesterone acetate 0.625/2.5 mg/day. Age was documented; weight, height, waist and hip perimeters as well as blood pressure were analyzed. Body mass index and waist-hip ratio were calculated. Glucose and lipid levels were measured. Statistical analysis was done with Student's test for independent samples for comparison among the groups. RESULTS: Thirty-three patients were studied, 17 were in the group without hormone therapy and 16 in the group with hormone therapy with conjugated equine estrogens and medroxyprogesterone acetate 0.625/2.5 mg/day were studied. When the groups were compared, the waist perimeter was significantly greater in the group without hormone therapy (98.7 +/- 7.8 cm vs. 92.9 +/- 9.1 cm, respectively, p < 0.05). No significant differences were found between the groups in blood pressure, glucose, total cholesterol, high-density lipoprotein cholesterol and triglycerides. Metabolic syndrome diagnostic criteria were completed in 12 patients of group 1 (70.6%) and 8 in group 2 (50%), without statistically significant difference between them. CONCLUSION: Hormone therapy group was associated to a lower abdominal perimeter and a lower waist-hip ratio. This let's suppose that hormone therapy has a beneficial effect on the prevention of metabolic syndrome. PMID- 19902627 TI - [Prenatal diagnosis ovarian cyst with an amputation at three months of age. A case report]. AB - In the neonate the pelvic masses regularly are usually benign until in 87%, those of ovarian origin most is of cystic and benign origin. CLINIC CASE: Of a pregnant of 26 years of age, primigesta, was detected to the fetus during the 24 weeks of gestation an abdominal cyst in pelvic hole of 3.5 diameter cm, non motive. The pregnancy culminated for caesarean operation, feminine product was obtained with weight when being born 3.400 kg. They were practiced 3 ultrasounds: when being born, to the month and two months of age, finding ovary cyst with the same characteristics. To the 3 months it is observed in 4th control ultrasound that the cystic image was no longer in pelvic hole, but it liberates in abdominal cavity for what decides to make exploratory laparotomy, where is a cyst of free amputated ovary in abdominal cavity of 3.5 x 4 diameter cm. The pathology results reported an ovary with necrosis areas and content cystic saculado. PMID- 19902628 TI - [Epithelioid uterine leiomyoma with giant cystic degeneration. A case report]. AB - At present, show woman 46-years-old, presented hiperpolimenorrea, pelvis ultrasound appearance with cystic giant mass in the uterine corpus; histopathological revealed a epithelioid leiomyoma with giant cystic degeneration. We considered it is the first reported case of this variant of the epithelioid leiomyoma. PMID- 19902629 TI - [Angiomyolipomas, tuberous sclerosis and pregnancy]. AB - Renal angiomyolipomas are uncommon tumours during pregnancy, but it's important to consider them in the differential diagnosis, for the increased risk of complications such as bleeding, specially during pregnancy. Their characteristics are different depending on if the tumour is associated or not with facomatosis. To show both possibilities, we describe two cases of pregnant women with angiomyolipomas: the first one presented with bleeding of the tumour (Wunderlich's syndrome), but surveillance without treatment was possible until 37th week of gestation, and embolization of the tumour was performed afterwards. The second patient had multiple well known angiomyolipomas, associated to tuberous sclerosis; even though she had required two previous embolizations, she had an uneventful pregnancy, without any episodes of bleeding. Both pregnancies ended successfully by means of a cesarean section. Since there are few papers available in the literature, there is not an agreement about therapeutic management when pregnant women present with bleeding angiomyolipomas (Wunderlich's syndrome). Treatment options include partial or total nephrectomy (with cesarean section simultaneously in patients at 28 weeks of gestation or later), transcateter selective arterial embolization (which can also be preventive), and sometimes, conservative management. If this last option is possible, there's still discussion about the risk of vaginal delivery compared with that of cesarean section, in terms of bleeding of the tumour. PMID- 19902630 TI - [Ovarian cystadenoma and ectopic pregnancy. A case report]. AB - A 40-year-old woman without symptoms, with a three-week amenorrhea, and with no previous pregnancy history. Three months before she followed treatment with clomifeno and no other risk factors. A measure of beta fraction is performed, finding a probable pregnancy of 3-4 weeks. A new measure of beta fraction is made three weeks later with an ultrasound, finding an important increment in measure, but without evidence of intrauterine pregnancy in the ultrasound. A new ultrasound is made seven weeks after her last period, finding a mass in the left ovary, but without increment in beta fraction. At week ten, she presents an uterine bleeding during a trip, which is diagnosed as a probable mole without any further medical treatment and from the 12th week, there is a considerable decrement in the beta fraction measure, without any other symptom, but the persistent adnexal mass at the left ovary, with irregular septum images at ultrasound. A laparotomy is performed finding an ectopic pregnancy surrounded by a serum cystadenoma. PMID- 19902631 TI - [Urethral erosion secondary to the placing of tension-free vaginal tape. A case report]. AB - Tension free vaginal tape is a minimally invasive surgical procedure for urinary incontinence treatment. Urethral erosion is an uncommon complication which presents in around 0.9% of the cases after the procedure. CASE: Female of 68-year old with an urodynamic diagnosis of type II stress urinary incontinence treated surgically with a tension free vaginal tape in February 2003 without any intraoperative complications. In the post-op she had acute urinary retention for one week, needing a transurethral catheter. Three months after surgery she reported dysuria, hematuria, frequency and urgency; the physical examination was normal with a positive urine culture to Escherichia coli sensitive to Nitrofurantoine. A cystoscopy was performed with the following findings: the tape was found in 25% of the right lower quadrant mid third of the urethra, the tape was cut vaginally without any further complications. PMID- 19902632 TI - [Deontology and moral in gynecology. 1953]. PMID- 19902633 TI - [Clinical practice guidelines. Diagnosis and treatment of primary dysmenorrhea in teenagers]. PMID- 19902634 TI - The Medical Society meeting with the insurance CEOs. PMID- 19902635 TI - Health care costs and economics 101: a vignette. PMID- 19902636 TI - A response to "Further thoughts on universal health care". PMID- 19902637 TI - HHS issues Interim Final Rule on health data breaches. PMID- 19902638 TI - The original desensitizers: strontium and potassium salts. AB - Many patients complain of teeth that are painful when exposed to a variety of harmless thermal and tactile stimuli. Sensitive tooth necks and root surfaces frequently are the unintentional by-products of aggressive oral hygiene practices and periodontal treatment. Dentists and the afflicted patients have resorted to many remedies for this common form of dental pain. In the past, many of these purported treatments were based on a fragmentary knowledge of the anatomic substrate and physiological processes underlying dentin sensitivity. Much progress has been made identifying dentin permeability and intradental nerve excitability as physiological parameters that can be modified by desensitizing agents. In this paper, rather than provide a comprehensive or critical review of desensitizing treatment, I will discuss the rationale and some of the history behind two early and popular classes of dentifrice-applied desensitizing agents; strontium and potassium salts. PMID- 19902639 TI - Advances in methodologies to characterize dentine and dentine hypersensitivity treatments in vitro. AB - The aim of this paper is to highlight recent advances in the development of analytical and in vitro methodologies to assess the potential efficacy of dentine hypersensitivity treatments. The paper will cover the utility of X-ray computed nanotomography, secondary ion mass spectrometry, dynamic laser scanning confocal microscopy, and x-ray microtomography in the context of characterizing dentine, understanding the permeability and mass transport properties of dentine, and exploring the mechanism of action for purported hypersensitivity treatments. It will be shown that X-ray tomography, secondary ion mass spectrometry, and dynamic fluid flow confocal microscopy provide unsurpassed information pertaining to the ultra-structural and mass transport properties of dentine, properties that underpin the hydrodynamic theory and the therapeutic route for desensitizing treatments. The paper concludes that advances in analytical instrumentation and in vitro methodologies have improved our understanding of the ultra-structural and chemical properties of dentine, mass transport within dentine, and the quality and predictive value of in vitro models. Such understandings aid the development of efficacious dentifrice formulations to treat dentine hypersensitivity. PMID- 19902640 TI - Development of an in situ methodology for the clinical evaluation of dentine hypersensitivity occlusion ingredients. AB - OBJECTIVE: The aim of these clinical studies was to evaluate an in situ dentine tubule occlusion model, and to determine the occluding effect from novel occluding agents on patent dentine tubules compared to a positive control (8% strontium acetate--Sensodyne Mint) and negative control (a non-occluding agent) after four days of brushing treatment. METHODS: These two in situ clinical studies were of single-center, randomized, crossover, single-blind design. Healthy participants wore two lower intra-oral appliances retaining four dentine samples for four treatment days for each period of the study. Samples were power brushed each day with the test product. Assessment utilized surface topological analysis with a replica-based methodology under scanning electron microscopy (SEM). RESULTS: Both clinical trials demonstrated that the positive control (8% strontium acetate) occluded dentine tubules significantly better (p = 0.0007; p < 0.0009) than the negative controls in the two studies, respectively. The experimental occluding agents demonstrated varying degrees of success for occluding effect compared to the controls. CONCLUSION: The methodology clearly demonstrates that this in situ clinical model can robustly and reproducibly detect the dentine tubular occlusive effects of positive and negative controls in the treatment of dentine hypersensitivity brushed on the dentine surface. Using this methodology, new occlusion agents for the relief of dentine hypersensitivity can be assessed for occlusive effects on dentine. PMID- 19902641 TI - Considerations for the development of over-the-counter dentifrices for the treatment and relief of dentin sensitivity. AB - There have been many recent articles and reviews covering the effectiveness of dentifrices for the treatment and relief of dentin sensitivity. To date, there are no articles which fully examine all the compromises that a dentifrice formulator and manufacturer have to make when developing an effective dentifrice technology in the context of theoretical approaches and proposed modes of action. This article provides a first review and discussion of the currently available technologies, nerve depolarization and occluding agents. The generalized formulation strategies and approaches of dentifrices containing these agents are discussed, demonstrating that simplified conclusions with regard to efficacy should be drawn with caution. PMID- 19902642 TI - Implications of noninvasive prenatal diagnosis. PMID- 19902643 TI - No longer need to RE-LY on Warfarin for stroke prevention? PMID- 19902644 TI - Waiting times for access, diagnosis and treatment in a cancer centre. AB - We analysed the waiting times for patients in a Dublin hospital from 2001 to 2006, and evaluated trends in each of 4 cancer diagnoses; breast, lung, colorectal and upper gastrointestinal (gastric and oesophageal). Measured times were; time from referral to first seen, time from first seen to diagnosis and time from diagnosis to treatment. Patient numbers increased 39% from 529 in 2001 to 737 in 2006. As a result waiting times have increased over the 6 years. While median time from referral to first seen for breast cancer was 7 days, it rose from 2 to 5 days for lung cancer, 1 to 2 days for colorectal cancer, and 1 to 6 days for upper GI cancers. The time from diagnosis to treatment rose from 8 to 15 days (breast), 10 days to 25 (lung), 6 to 14 days (colorectal) and 7 to 13 days (Upper GI). Waiting times however, remain within international standards. PMID- 19902645 TI - Blastocyst transfer for multiple prior IVF failure: a five year descriptive study. AB - Patients with recurrent IVF failure are generally regarded as having a poor prognosis, and when female age exceeds 35 yrs such patients face a particularly bleak outlook. This study reported on blastocyst transfer (BT) performed over a five-year interval in patients seeking "second opinion" after multiple failed IVF cycles. Clinical features and reproductive outcomes were compared between two sets of poor-prognosis IVF patients undergoing BT for the first time, the initial group underwent treatment in 2002 (n=66) and a second group presented five years later (n=392). The two clinical sets had no patients in common. The 2002 group had an average of 3.5 (+/- 1.1) prior failed IVF cycles at baseline, and mean (+/ SD) patient age was 36.4 (+/- 3.9) yrs. Average number of oocytes retrieved in this group was 10.4 (+/- 5.3) with a fertilisation rate of 58.8%. Although embryo arrest resulted in no transfer for 19 patients (28.8%), clinical pregnancy was achieved for 59.6% of transfers. Five years later, 392 patients underwent BT, but this group had an average of 4.5 (+/- 2.3) prior failed IVF cycles. Mean (+/- SD) female age was 36.0 (+/- 3.9) yrs, and the average number of oocytes retrieved in this group was 9.1 (+/- 5.4); the fertilisation rate was 59.5%. No blastocysts were available for transfer in 99 cases (25.3%); clinical pregnancy was achieved for 50.0% of transfers. The number of blastocysts transferred was similar in the two groups (1.6 vs. 1.3; p=0.06); the twinning rate rose slightly from 8.2% to 15.1% (p=0.12) despite an increased utilisation of single embryo transfer in 2007 (19.7% vs. 22.2%; p=0.40). Comparisons from 2002 and 2007 found no important differences between the two patient groups, except for a significantly higher rate of prior failed cycles in the 2007 group (p<0.001). This refractoriness was accompanied by a somewhat reduced blastocyst cryopreservation rate in 2007, compared to 2002 (27.6% vs. 29.5%; p=0.44). Clinical pregnancy rates are not adversely affected by application of BT in patients with multiple prior unsuccessful IVF cycles. For these patients, our data suggest that extended embryo culture and BT should be considered. Further controlled studies are needed to document more precisely the role of BT in this sub-set of refractory IVF patients. PMID- 19902646 TI - Chronic kidney disease in general practice: prevalence, diagnosis, and standards of care. AB - There is little prevalence data for chronic kidney disease (CKD) in Ireland and it has been suggested that rates of diagnosis of CKD in primary care are low. The aim of this cross sectional study was to examine the prevalence, diagnosis and standards of care for CKD. All patient records in three general practices in the West of Ireland were reviewed. In 2602 patients > 50 years in the community, 435 (16.7%) had chronic kidney disease defined as eGFR <60 ml/min/1.73 m2. Of these 435 individuals, only 58 (13.3%) had a diagnosis of CKD documented in their patient record. Among all patients with an eGFR <60 ml/min/1.73 m2, those with a documented diagnosis of CKD were significantly more likely to be prescribed an ACE/ARB and a lipid-lowering agent and were more likely to have had an ACR/PCR checked in the previous twelve months. Blood pressure was being appropriately monitored in the majority of patients but irrespective of eGFR level or a documented diagnosis of CKD, less than a fifth of patients had achieved a target of <130/80 mmHg. CKD is common in primary care but remains largely undiagnosed and blood pressure control remains suboptimal. A key step in improving care appears to be documenting the diagnosis which in turn appears to lead to improved standards of care and risk factor management. PMID- 19902647 TI - Inter-hospital transport of critically ill children. AB - Most Irish paediatric tertiary care services are centralised in Dublin. Many children are transferred there each year from regional paediatric units around the country. We aimed to quantify and describe all children transferred from one regional tertiary hospital over a two year period. Seventy three out of 75 identified transfers were examined. Sixty nine transfers (94.5%) were sent to the major tertiary centre. Fifteen (20.5%) required intensive care services for transfer. Seventeen seriously ill neonates required transfer, however only 4 (23.5%) of those met both the criteria for and the availability of the National Neonatal Transfer Team (NNTT). Significant events during transfer were only documented in 3 cases. Most transfers arrived in Dublin outside normal working hours. Standards of documentation were found to be very inconsistent. In conclusion, a national transport service for all critically ill children is urgently needed in Ireland. PMID- 19902648 TI - Complete traumatic laryngo-tracheal separation. AB - Laryngotracheal separation injuries are rare and potentially fatal. Immediate respiratory signs may include dysphonia, aphonia, hemoptysis, subcutaneous emphysema and a sucking wound. Patients with this injury usually die at the site of the trauma. The absolute life saving intervention for patients with laryngotracheal injury is airway control via routine intubation or emergency tracheostomy. We present an extremely rare case of complete laryngotracheal separation in a teenager driving a quad bike in a 'clothes line' type injury with chicken wire. This case highlights the need for prompt airway evaluation, radiological imaging required, surgical management and long term injury sequelae. PMID- 19902649 TI - A pernicious leucoencephalopathy. AB - Pernicious anaemia may manifest various neurological symptoms and signs ranging from the subtle to the dramatic. We describe a young man with cobalamin deficiency presenting with sensorimotor deficits, ataxia, dysarthria, mild cognitive deterioration and altered mood of insidious onset. The MRI brain findings were in keeping with a leucoencephalopathy without evidence of MRI changes in the spinal cord. This constellation of features has been reported rarely. His response to treatment as well as the marked improvement of the leucoencephalopathy on imaging suggests at least partial reversibility of the neurological deficits. PMID- 19902650 TI - Multicentric Castleman's disease & HIV infection. AB - We report the case of a 35 year patient from Nigeria who presented with fever and splenomegaly. The initial diagnosis was Salmonellosis. However, relapsing symptoms lead to a re-evaluation and ultimately a diagnosis of Multicentric Castleman's Disease (MCD). There is no gold standard treatment but our patient responded to Rituximab and Highly active anti-retroviral therapy. MCD is a rare, aggressive disease that should be considered in a HIV positive patient presenting with fever and significant lymphadenopathy. PMID- 19902651 TI - Radiographic appearance of a post-epidural headache. AB - We report the case of a 35-year-old lady who presented with a 6-day history of a postural headache following an uncomplicated epidural catheter insertion. Meningitis was initially suspected and a neurology review was obtained. CT and MRI brain revealed features suggestive of meningitis. However these radiological features are also consistent with post dural puncture headache (PDPH). This case highlights the under reported and possible misleading radiographical features of PDPH. PMID- 19902652 TI - Transplantation, multi-organ donation & presumed consent: a 3 year survey of university students. AB - We profile the practises and attitudes of university students in Ireland towards consent for organ donation. 1103 students were surveyed. Only 34.6% (382/1103) carried organ donor consent cards, although the majority were favourably disposed towards donation. Only 9% (96/1103) were against donation. In regard to presumed consent only 38% (177/470) were in favour of changing the current "opt-in" consent methodology to presumed consent. These findings show a favourable opinion towards donation among Irish university students. However this may result in few actual donations in the event of brain death, as the majority do not carry donor cards and do not want to change to a presumed consent regime. The most common answer for not carrying a card was that the individual had not formalised a decision. Mandated choice at a fixed point could significantly reduce this ambivalence. PMID- 19902653 TI - Prescribing sugar-containing medicines for children--are we forgetting "primum non nocere"? PMID- 19902654 TI - Endoscopic ultrasound and EUS-guided FNA in the diagnosis of rectal endometriosis. AB - Endometriosis is characterised by the presence of endometrial glands and stroma outside the uterus. The GI tract is the most common site for extra-pelvic endometriosis, in particular the rectum and sigmoid colon. Using endoscopic ultrasound (EUS), which combines endoscopy with real-time ultrasonography, the wall of the GI tract and adjacent structures can be examined. EUS-guided fine needle aspirates can be also obtained during the procedure. We report a case of rectosigmoid endometriosis which was diagnosed using EUS. PMID- 19902655 TI - Prisoners with mental disorders in Europe. PMID- 19902656 TI - Reliability and limitations of the durometer and PressureStat to measure plantar foot characteristics in Native Americans with diabetes. AB - Neuropathy with excessive weight-bearing activity may lead to foot changes that place it at risk for ulceration. Information about instruments to measure plantar skin hardness and pressures of the foot in adult Native Americans with diabetes is presented. Skin hardness was measured at 10 sites (plantar side of the hallux, third and fifth toes, first, third, and fifth metatarsal heads, medial and lateral midfoot, heel, and the dorsal aspect between the hallux and second toe) with a durometer. The PressureStat yielded data about barefoot, static pressures. The durometer was shown to be reliable (r = .62 to .91) at all sites of the foot except at the third and fifth toe, the medial midsection of both feet, and at the third metatarsal head and between the hallux and second toe, the dorsal aspect of the left foot. With four raters, concordance was found to be acceptable at the hallux, the third toe, the first, third, and fifth metatarsal heads and the heel (W = .61 to .86) for the PressureStat. Since most diabetic foot ulcers occur at the hallux, first, third, and fifth metatarsal heads, both the PressureStat and durometer may be reliable screening tools to determine the degree of risk. PMID- 19902657 TI - Support for the reliability and validity of a six-item state anxiety scale derived from the State-Trait Anxiety Inventory. AB - Identifying the most efficient and theoretically appropriate methods to assess patient anxiety in fast-paced medical environments may be beneficial for clinical purposes as well as for research. The purpose of this study was to examine the reliability and validity of two previously published six-item versions of the State form of the State-Trait Anxiety Inventory (STAI) and to identify the version that would be most appropriate to use with a sample of parents who had infants with normal or abnormal newborn screens. In the current study, confirmatory factor analyses were conducted to evaluate the fit of the two six item forms with STAI data collected at three time points from 288 parents of 150 infants. Study groups of parents were based upon infant newborn screens and subsequent diagnostic testing to include cystic fibrosis (CF; n = 26), congenital hypothyroidism (CH; n = 39), CF Carriers (CF-C; n = 45), and healthy infants (H; n = 40). The results showed the version containing items 1, 3, 6, 15, 16, and 17 of the State form of the STAI to be a better fitting model across all three time points, and it had better internal consistency than the version containing items 5, 9, 10, 12, 17, and 20. Both short forms were highly correlated with the 20 item STAI score, and all internal consistency reliabilities were greater than .90. It was concluded that the version containing items 1, 3, 6, 15, 16, and 17 of the State Anxiety scale was a reliable and valid instrument for this study sample. PMID- 19902658 TI - Psychometric properties of the Social Comparison Motives Scale. AB - This article describes the 19-item Social Comparison Motive Scale [SCMS], a measure of adolescents' motives for social comparison related to pregnancy. Dimensions and items were developed based on adolescent focus groups. The instrument was reviewed for content validity, pilot tested, and administered to 431 adolescents aged 14-18 years. Principal axis factor analysis with oblique rotation supported five dimensions. Convergent and discriminant validity were demonstrated by moderate correlations (r = .50) between the SCMS and the Iowa Netherlands Comparison Orientation Measure and low correlations (r = .15) between the SCMS and the Rosenberg Self-Esteem Scale. Cronbach's alphas were .91 overall and .71 to .85 for the subscales. The SCMS demonstrated reliability and validity as a measure of adolescents' motives for comparing themselves with others about pregnancy. PMID- 19902659 TI - Development of the Military Women's Attitudes Toward Menstrual Suppression Scale: from construct definition to pilot testing. AB - The Military Women's Attitudes Toward Menstrual Suppression scale (MWATMS) was created to measure attitudes toward menstrual suppression during deployment. The human health and social ecology theories were integrated to conceptualize an instrument that accounts for military-unique aspects of the environment on attitudes toward suppression. A three-step instrument development process was followed to develop the MWATMS. The instrument was pilot tested on a convenience sample of 206 military women with deployment experience. Reliability was tested with measures of internal consistency (alpha = .97); validity was tested with principal components analysis with varimax rotation. Four components accounted for 65% of variance: Benefits/Interest, Hygiene, Convenience, and Soldier/Stress. The pilot test of the MWATMS supported its reliability and validity. Further testing is warranted for validation of this instrument. PMID- 19902660 TI - Reliability and validity of the Jefferson Scale of Empathy in undergraduate nursing students. AB - Evidence has been reported in support of the reliability and validity of the Jefferson Scale of Physician Empathy (JSPE) when used with physicians, medical students, and nurses. This study examined the psychometrics of a modified version of the scale in undergraduate nursing students. The modified version of the JSPE was administered to 333 nursing students at different levels of training. Three underlying constructs, that is, "Perspective Taking," "Compassionate Care," and "Standing in Patient's Shoes" emerged from the factor analysis of the scale that were consistent with the conceptual framework of empathy, thus supporting the construct validity of the scale. The coefficient alpha was .77. Scores of the empathy scale were significantly correlated with the scores of the Jefferson Scale of Attitudes toward Physician-Nurse Collaboration (r = .38, p < .001). Women scored higher than men, and those with more clinical experiences scored higher than others. It was concluded that the empathy scale used in this study is a psychometrically sound instrument for measuring empathy in undergraduate nursing students. PMID- 19902661 TI - [Opinion articles: a new series for 2010]. PMID- 19902662 TI - [Perioperative myocardial infarction]. PMID- 19902663 TI - Antiarrhythmic and arrhythmogenic action of inosine in experimental ventricular tachyarrhythmias. AB - OBJECTIVE: To study the possible action of inosine on experimental ventricular tachyarrhythmias. MATERIAL AND METHODS: We used 92 mongrel dogs weighing 13 kg-17 kg, anesthetized with 30 mg/kg sodium pentobarbital applied intravenously. Myocardial lesions were induced by injecting 1 ml-1.5 ml of 70% phenol in the free wall of the left ventricle. In 36 dogs, the ventricular arrhythmia (VT) was induced 30 min later with aconitine crystals inserted into the periphery of the damaged area; in 16, VT was due only to myocardial damage and in the other 13 VT was spontaneously originated. Twenty-nine animals constituted the control group; no inosine was administered to them. The possible effects of inosine were studied in 63 animals. Leads II, aVR or aVL, right and Left unipolar intraventricular leads and that on the wall of the superior vena cava were recorded under control conditions, once the myocardial damage had been induced, during the ventricular tachycardia, and following the injection of inosine. Of the 63 inosine-treated animals; in 34, VT was due to aconitine; in 16, it was produced only by the myocardial damage and, in 13, VT was presented spontaneously. RESULTS: Sinus rhythm was not reestablished in the animals of the control group. Inosine reestablished the sinus rhythm in 26 of 34 dogs (76%) that received phenol and aconitine, in 13 of the 16 (81%) presenting only the myocardial damage, and in 6 of the 13 (46%) with spontaneous ventricular tachycardia. In some experiments, inosine induced supraventricular tachycardias, ventricular-atrial blocks, and ventricular pre-excitation phenomena. CONCLUSIONS: In this experimental series, inosine showed antiarrhythmic and arrhythmogenic effects, similar to those of adenosine from which it derives. PMID- 19902664 TI - Antiarrhythmic and cardioprotective effects of remifentanil in anesthetized dogs. AB - OBJECTIVE: To study the antiarrhythmic effect of remifentanil in experimental arrhythmias in dogs. METHODS: We used dogs weighing 12 kg-18 kg anesthetized with 30 mg/kg sodium pentobarbital given intravenously. Ventricular arrhythmia, ventricular fibrillation and death were induced with digoxin (9 microg/kg/min). In another model, two types of arrhythmia were induced in the right atrium, one of them with aconitine crystals placed on the right atrium and the other was induced in the basement of the right atrium by electrical stimulation. The potential antiarrhythmic action of remifentaniL was investigated in ventricular and atrial arrhythmias by the administration of an intravenous bolus after toxic signs were evident. Thus, two arrhythmias with different mechanisms were generated. Leads DII, unipolar left intraventricular and right atrial leads, and left ventricular pressure were used to record control tracings and tracings in presence of remifentanil, during ventricular arrhythmia. RESULTS: Remifentanil abolished toxic effects of digoxin, it eliminated the A-V dissociation and ventricular extrasystoles, reverting to sinus rhythm in each case. Remifentanil extended the time to reach lethal doses from 63.25 +/- 11.3 to 100 +/- 11.8 min. These effects were blocked by naloxone (0.01 microg/kg) applied before remifentanil. In the two arrhythmias model, remifentanil suppressed both, ectopic focus and atrial flutter. CONCLUSIONS: Remifentanil elicits antiarrhythmic and cardioprotective effects in experimental ventricular arrhythmias induced by digoxin and in a model of two atrial arrhythmias induced by aconitine and by electrical stimulation. PMID- 19902665 TI - [Diagnosis of perioperative myocardial infarction within the first seventy-two hours after cardiac surgery]. AB - One of the most frequent complications of cardiac surgery is the perioperative myocardial infarction (PMI). Incidence of PMI shows a wide variation because an accurate detection of this complication is difficult in the early postoperative stage. The objectives of the present study were to determine in our population of patients the incidence of PMI during the first seventy two hours after cardiac surgery as well as associations among the accepted criteria to diagnose this complication. PATIENTS AND METHODS: One hundred sixty four adults patients undergoing elective cardiac surgery were studied. With baselines preoperative studies, serial electrocardiographic, enzymatic [(determination of serum creatine kinase isoenzyme (CK-MB)] and echocardiographic studies were performed during the first 72 hours after cardiac surgery. Diagnosis of PMI was established with two or all the three positive criteria [electrocardiographic (ECG), enzymatic (CK-MB) and echocardiographic (ECHO)]. RESULTS: In 24 (15%) patients PMI was diagnosed. In this group 13 (54%) all the three criteria were positive. In 8 (33%) patients CK-MB and ECHO were positive. In 3 (13%) patients CK-MB and ECG were positive. CONCLUSIONS: In our population the incidence of PMI (15%) is agree with the reported in previous studies. In most of cases of PMI all the three diagnostic criteria are positive. When diagnosis is established only with two criteria, in most of cases these are CK-MB and ECHO. PMID- 19902666 TI - [Prevalence of syncope in a sample of Mexican women residents in Mexico City]. AB - OBJECTIVE: To identify the frequency of syncope in a sample of women regular residents of Mexico City. METHODS: Two hundred and twenty one women between 18 and 88 years old with voluntary participation were included in the study. Trained interviewers through a structured questionnaire obtained syncope information. Prevalence of syncope was obtained, and it's relation with: age, time since the last syncope and frequency of syncope in their live span was registered. RESULTS: Thirty eight percent informed they had suffered syncope, of them 50% had only one syncopal episode during their lives. The others had two or more episodes. Most women had their last syncope between 19 and 50 years of age. CONCLUSIONS: The frequency of syncope was a little higher than previous studies. This is a first approximation, which must be corroborated with larger studies with well population variability representation. PMID- 19902667 TI - [Syncope in the elderly: findings of diagnostic complementary tests in a University Hospital]. AB - Syncope is a common symptom among older adults. Its aetiologic diagnosis is mainly clinical, but when it has an unknown origin, complementary studies are necessary. We present the experience of a single center in diagnosing Unknown Origin Syncope (UOS). METHODS: We performed a transversal retrolective study to assess the different tests done to patients studied because of USO. RESULTS: The mean age of the older group was 77.6 +/- 6.9 years. The 24-hour Hotter were abnormal in 77.9% of patients, but diagnostic in only 16.3%. Age older than 65 years was associated with a 1.9 increase (CI 95% 0.9-4) in the possibilities of having a diagnosis, while age lesser than 65 was associated with a 0.5 risk CI 95% 0.2-1.08 of having a diagnosis. The 48-hour holter showed similar results. Older age was associated with a 1.69 OR, CI 95% 0.6-4.4 to have a positive Tilt test (91% of older patients). They had mainly the vasodepressor variety of neurally-mediated syncope. (57.1% Vs 40.1%, p = 0.01). Of the 6 electrophysiologic studies performed in people older than 65 years, 3 where diagnostic (50% Vs 33.3%, p = 0.6). CONCLUSIONS: Complimentary tests in people elder than 65 years are diagnostic in a higher proportion than in their younger counterparts. Nevertheless there is a higher risk of false-positive results, so tests such as head-up tilt test should be more carefully interpreted. PMID- 19902668 TI - [Psychopathology and personality in patients with vasovagal syncope]. AB - Syncope is characterized by a sudden and brief Loss of the normal state of consciousness (fainting), caused by multiple factors (biological, psychological and social), which consequences can be Lethal if it is not timely diagnosed and treated. These episodes affect the total sphere of the subject (on an individual, work, school, social, and family level). PURPOSE OF THE STUDY: Contribute to the study of vasovagal syncope (WS) patients, describing their personality and psychopathological characteristics. METHOD: Minnesota Multiphasic Personality Inventory (MMPI) was used to evaluate psychopathology in a sample of 30 subjects diagnosed with (WS), attended in the National Institute of Cardiology "lgnacio Chavez", making a descriptive analysis of the complete sample, regarding demographic variables and inventory scores. RESULTS: The scales with higher scores found within these subjects were hypochondriasis (= 67.43), depression (= 69.83), hysteria (= 67.83) among others. CONCLUSIONS: In general, patients with (WS) show significant levels of anxiety and depression, dissatisfaction, bad mood, pessimism, concern, somatic complaints, difficulty to solve problems adaptively, among others. This study remarks the importance of knowing these characteristics to implement treatments that manage this disease properly and may improve patients quality of life, as well as their physical and mental health. PMID- 19902669 TI - [Case report: idiopathic ventricular tachycardia from the aortic sinus cusp]. AB - Idiopathic ventricular tachycardia is identified in 10% of the patients presenting with ventricular tachycardia, and they consist of various subtypes that can originate from different areas, including the aortic cusps which represent 0.7% of the total. Electrocardiographically, these ventricular tachycardias display a left bundle branch block pattern and inferior axis, and although rare, should be considered in the differential diagnosis with tachycardias originating from the septal region of the right ventricular outflow tract, which comprise 80% of the idiopathic ventricular tachycardias. Despite the vicinity of the left coronary artery ostium, radiofrequency catheter ablation can be curative in more than 90% of cases with a low risk (< 1% of serious complication). Therefore, it must be considered first-line therapy in patients who have failed or are intolerant to therapy with antiarrhythmic agents. The aim of this article is to describe the first case reported in Mexico of a successful ablation idiopathic ventricular tachycardia from the aortic sinus cusp in a patient with incessant ventricular tachycardia. PMID- 19902670 TI - [Anatomic support in the echocardiographic diagnosis of right atrioventricular connection]. AB - OBJECTIVE: To show the close relationship between the anatomic features and the echocardiographic image in the absence of right atrioventricular connection. MATERIAL AND METHOD: Echocardiographic study of a patient with right atrioventricular connection and a corresponding anatomic specimen from the Museum of Embryology of the INC "lgnacio Chavez". RESULTS: A close correlation between an anatomic specimen with absence of right atrioventricular connection and an echocardiographic image of a patient with the same cardiopathy was established. CONCLUSION: The correlation between the anatomic features and the echocardiographic image, supports the imagenologic diagnostic precision of this method. PMID- 19902671 TI - [Remote monitoring of automatic implantable cardiovascular devices (pacemakers, implantable cardioverter defibrillator and cardiac resynchronization)]. AB - In the era of communication technology, new options are available to monitor patients with Automatic Implantable Cardiovascular Devices (AICD) implanted pacemaker (PM) and Automatic Implantable Defibrillators (AID) and Cardiac resynchronization system (CRS). Most companies offer devices with wireless capabilities to communicate automatically with transmitters, allowing remote monitoring device. These systems are being widely used in USA for remote monitoring and have been introduced more recently in Europe, where adoption is increasing. There have also been introduced some systems in Latin America; Mexico in particular has this type of monitoring. This article describes the systems currently existing, available data in the literature in relation to its monitoring and surveillance of the automatic implantable cardiovascular devices (DAIC) and finally, discuss some unresolved issues. PMID- 19902672 TI - [Iberoamerican Cardiovascular Journals. The vital need for cooperation]. PMID- 19902673 TI - [Gynecology and obstetrics]. PMID- 19902674 TI - [Levels of estradiol and serum testosterone in healthy postmenopausal women and its relationship with symptoms]. AB - BACKGROUND: Several studies have tried to evaluate in which estradiol (E2) and testosterone (T) levels the climacteric symptoms appear, but this has not been possible at all. OBJECTIVE: To evaluate in which E2 and T levels climacteric symptoms appear, as well as its relation with T/E2 ratio. MATERIAL AND METHODS: Twenty nine healthy climacteric women [perimenopausal (n = 13) or postmenopausal (n = 16)] which have not received nor were receiving hormone therapy were studied. Age, body mass index, waist hip ratio and menstrual status were documented. An analog visual scale was used to evaluate the intensity of 13 climacteric symptoms. A blood sample was taken for the measurement of E2 and T measurement by radioimmunoassay. Pearson's correlation analysis was done between the sum of scores of all climacteric symptoms and with the score obtained from each of the climacteric symptoms and E2, T levels and T/E2 ratio. All patients gave their consent to participate in the study. RESULTS: The average age of women was 49.9 +/- 5.1 years and the time since menopause 43.9 +/- 53.2 months. The sum of the magnitudes of the symptoms was 71.2 +/- 24.9. The average level of E2 and T was 65.5 +/- 101.1 pg/ml and 14.5 +/- 10.7 ng/ml respectively. No correlation was observed between the sum of the magnitudes of the symptoms neither with the magnitude of each symptom with E2 and T levels and T/E2 ratio. CONCLUSIONS: It was not possible to determine with precision in which E2 and T levels climacteric symptoms appear and T/E2 ratio also was not useful for this purpose. PMID- 19902675 TI - [Intrauterine misoprostol for the prevention of bleeding cesarean]. AB - OBJECTIVE: To evaluate efficacy of misoprostol by intrauterine route for the prevention of the obstetrical hemorrhage and to know its effects collaterals. PATIENTS AND METHODS: Clinical and randomized test with 200 patients to those who one took place to them Caesarean. The Group A was applied placebo and the group B misoprostol (group B; 800 microg) intrauterine, after the birth of its newborns. The necessity of additional uterotonics was compared, loss of hemoglobin and hematocrit, and the effects collaterals. RESULTS: In group B it diminished the necessity of additional uterotonics to 50% and the loss of hemoglobin in 39.6%. In the group greater loss of 3 was registered g of hemoglobin in 13 versus 3% of the patients. With respect to the hematocrit, its loss was reduced in 40.6%. The patients in those who the hematocrit in more fell than 10%, registered themselves in 7 versus 1% cases. The effects collaterals were few and not so serious. CONCLUSION: The intrauterine combination of misoprostol and oxitocin diminishes the postcaesarean sanguineous loss and has brings about few effects collaterals. PMID- 19902676 TI - [Non-hormonal treatment for vasomotor symptoms during menopause: role of desvenlafaxine]. AB - OBJECTIVE: To review the published data that include Selective Serotonin Reuptake Inhibitors (SSRIs) and Serotonin-Norepinephrine Reuptake Inhibitors (SNRIs), especially desvenlafaxine, on vasomotor symptoms (VMS). METHODS: This review is based on published data in Medline (1990-2009) for studies on SSRI and SNRIs, especially desvenlafaxine, and VMS. RESULTS: There is increasing evidence that both norepinephrine and serotonin are associated with the communication and modulation of the temperature homeostasis maintained by the hypothalamus. Different studies demonstrated the modest efficacy of SSRIs and SNRIs on VMS. Recently, a program of clinical trials with desvenlafaxine (a salt from the major metabolite of venlafaxine) has been developed for VMS. Currently, there are three randomized, double-blind clinical trials published, showing a significantly higher efficacy of desvenlafaxine versus placebo on VMS. There were also increased minor side effects with desvenlafaxine, especially nausea, at the beginning of the treatment. CONCLUSIONS: A non-hormonal alternative- desvenlafaxine--has proven efficacy for VMS. There was also an increase in minor side effects, especially nausea, at the beginning of the treatment. There are clear subgroups of patients with VMS eligible, such as women with hormone dependent cancers, women who do not want to be treated with hormone therapy or just want to get relief of their vasomotor symptoms. PMID- 19902677 TI - [Hormonal therapy in metastatic breast cancer]. AB - The primary objective in metastatic breast cancer is tumor control and symptom palliation. Factors to be considered are: efficacy, tolerance and quality of life as well as patient preferences. In the Hormone Receptor Positive Group, Hormonal treatment is the best choice because of it's effectiveness and good toxicity profile. Endocrine therapy has two main targets: the first one is to block estrogen production. In premenopausal women this can be through ovarian ablation. In postmenopausal women this is achieved by blocking the peripheral conversion of androgens to estrogens by blocking the enzyme known as aromatase. The other option is to block the action of the estrogen on it's receptor with the group of drugs known as selective estrogen receptor modulators (SERM). This class of drugs can be used in pre and postmenopausal women. Treatment should be tailored according to patient characteristics and menopausal status. PMID- 19902678 TI - [Aggressive angiomyxoma of the vulva. Case report and literature review]. AB - Aggressive angiomyxoma is a rare vulvovaginal, perineal or pelvic mesenchymal neoplasm with a marked tendency to local recurrence but does not metastasize. A case of an aggressive angiomyxoma of vulva in a 39-years-old women with an illness of one year prior to examination, with a slow and progressive growth of the left vulvar region, without other symptoms. During physical examination, a piriform tumor of 15x10 cm was found, located on the left labia majora, soft tissue dependent. Wide resection of the tumor were performed. Hystopathology reported an aggressive angiomyxoma of the vulva, with tumor in resection margins. The patient was treated with a 65Gy postsurgical radiotherapy and gosereline 3.6 mg monthly, during 6 cycles. Aggressive angiomyxoma is a rare neoplasm 150 cases has been reported. The treatment is surgical resection. Radiotherapy and hormonal adyuvant is not fully stablished. PMID- 19902679 TI - [Enterobiasis vaginal. Report of one case]. AB - The nematode Enterobius vermicularis is normally found within the human gastrointestinal tract. Pregnant females migrate towards the anus to lay their eggs. During this nocturnal migration some worms find another adjacent orifices, most commonly the female genitourinary tract. The presented case is a 34-year-old woman referred to the Colposcopy Unit of the Antiguo Hospital Civil of Guadalajara Fray Antonio Alcalde by cervical erosion. With the colposcopy a live worm in the vagina was visualized; it was identified as a gravid female of Enterobius vermicularis by microscopy. The diagnosis of ectopic enterobiasis was an incidental finding during the colposcopic examination of the patient with diagnosis of ectopy. PMID- 19902680 TI - [When, why and how to change abnormal fetal position during pregnancy]. PMID- 19902681 TI - [Clinical practice guidelines. Diagnostic and management of stress urinary incontinence]. PMID- 19902682 TI - Medication therapy management--not just for seniors anymore. PMID- 19902683 TI - Medicare advantage can, indeed, be superior. PMID- 19902684 TI - Improved adherence highlights specialty pharmacy's potential. PMID- 19902685 TI - Majority of docs contract with managed care. PMID- 19902686 TI - Large PBMs transform old business models. PMID- 19902687 TI - Questioning The reform agenda. How poverty affects cost and outcomes. Interview by John Marcille. PMID- 19902688 TI - Docs and insurers work to advance health literacy. PMID- 19902689 TI - Insurers slow to adopt social media practices. PMID- 19902690 TI - Hypertension epidemiology and economic burden: refining risk assessment to lower costs. AB - BACKGROUND: Hypertension (HTN) continues to be a serious public health problem in the United States and is a major risk factor for stroke, heart failure, myocardial infarction, and other serious cardiovascular and renal diseases. Because HTN can be asymptomatic, its detection and control continues to be a challenge. The total economic burden of HTN is estimated at $73.4 billion in 2009. OBJECTIVE: To examine the potential prognostic utility of biomarkers to assess hypertension-related cardiovascular risk and their potential impact on treatment in the context of current epidemiology and demographics of HTN. SUMMARY AND CONCLUSIONS: Although blood pressure control rates among people treated for HTN have increased from 51.3 percent to 63.9 percent over the past five years, there remains a vast unmet need for improved efficiency and effectiveness in diagnosis and treatment. Biomarkers provide a promising approach to improve detection and management of disease progression while optimizing health care expenditures. PMID- 19902691 TI - New genomic cancer test allows better decisions. PMID- 19902692 TI - Middle class faces steeply rising premiums. PMID- 19902693 TI - Education--leadership--collaboration. PMID- 19902694 TI - "Lean into it" in 2006. PMID- 19902695 TI - Self-care needs of cataract patients following ambulatory surgery. AB - Cataract patients are discharged from ambulatory surgery centers soon after surgery and must be able to provide postoperative self-care at home. It is important to identify what patients perceive as their needs for self-care in order to provide the information they need in the limited time available. During the review of literature, no studies were found that looked at the needs of these patients. The Patient Learning Needs Scale, developed by Galloway et al, was used in this descriptive research study, guided by Orem's Self-Care Framework, to determine the perceived needs of patients for self-care following cataract surgery in an ambulatory surgery center. The subscales of Complications and Symptoms, followed by Medications, were the primary areas identified by cataract patients. This has implications for improving patient education, patient satisfaction, and self-care for these patients. PMID- 19902696 TI - Let's get it straight: the nursing implications of adjustable sutures in the adult strabismus patient. AB - The objective of this paper is to highlight the important nursing interventions and clinical implications in the care of the strabismus patient treated with adjustable sutures. Nurses must use their assessment skills in monitoring the strabismus patient for vagal responses, bleeding, and anxiety during the suture adjustment period. Following adjustment, pain, nausea, vomiting, infection, and safety should be monitored and treated appropriately. Awareness of these nursing implications and human behavioral responses empowers the ophthalmic nurse to provide appropriate and effective care to the strabismus patient treated with adjustable sutures. PMID- 19902697 TI - Video compact discs for patient education: reducing anxiety prior to cataract surgery. AB - For this controlled trial, 463 patients were randomly assigned to two groups. The control group received cataract information leaflets and the study group received both leaflets and an educational video compact disc (VCD). Eighty percent of the subjects were more than 60 years old, and 86% were Chinese. Both groups showed significant knowledge acquisition and were marginally less fearful and nervous than before the study began. Between groups, however, there was no significant difference in knowledge and anxiety level. The subjects' preferences of information formats were: verbal (spoken) 54%, VCD 47%, and printed 36%. PMID- 19902698 TI - Pupil dilation and driving: guidelines for ophthalmic nurses and technicians. AB - Ophthalmic nurses and technicians often instill the dilating eyedrops before patients undergo ocular examination. These drops may cause some problems for patients who will be driving afterward. As professional health care providers, we have a duty to inform patients not only before the appointment but also prior to the instillation of the dilating drops that their vision may be altered due to the dilating eyedrops. It is extremely common for patients to have their pupils dilated when attending an ophthalmic outpatient appointment. This is to aid in the examination of the lens, vitreous, and fundus. The most common eyedrop used for pupil dilation in tropicamide 1%. This produces mydriasis and a small amount of cycloplegia. Mydriasis can last for several hours after instillation. PMID- 19902700 TI - Auditory and visual distraction. PMID- 19902699 TI - Laser and surgical treatments for glaucoma: an overview. PMID- 19902702 TI - Auditory fitness for duty: a review. AB - BACKGROUND: Auditory fitness for duty (AFFD) refers to the possession of hearing abilities sufficient for safe and effective job performance. In jobs such as law enforcement and piloting, where the ability to hear is critical to job performance and safety, hearing loss can decrease performance, even to the point of being hazardous to self and others. Tests of AFFD should provide an employer with a valid assessment of an employee's ability to perform the job safely, without discriminating against the employee purely on the basis of hearing loss. PURPOSE: The purpose of this review is to provide a basic description of the functional hearing abilities required in hearing-critical occupations, and a summary of current practices in AFFD evaluation. In addition, we suggest directions for research and standardization to ensure best practices in the evaluation of AFFD in the future. RESEARCH DESIGN: We conducted a systematic review of the English-language peer-reviewed literature in AFFD. "Popular" search engines were consulted for governmental regulations and trade journal articles. We also contacted professionals with expertise in AFFD regarding research projects, unpublished material, and current standards. RESULTS: The literature review provided information regarding the functional hearing abilities required to perform hearing-critical tasks, the development of and characteristics of AFFD protocols, and the current implementation of AFFD protocols. CONCLUSIONS: This review paper provides evidence of the need to institute job-specific AFFD protocols, move beyond the pure-tone audiogram, and establish the validity of test protocols. These needs are arguably greater now than in times past. PMID- 19902701 TI - Examining the effectiveness of traditional audiological assessments for nursing home residents with dementia-related behaviors. AB - BACKGROUND: It is currently estimated that the resident population of individuals over the age of 65 living in nursing homes will double by 2020. Nearly one-third of all nursing home residents have difficulty seeing or hearing, 46% have some form of dementia, and 30-84% of those with dementia in nursing homes show some form of agitation. Nursing home residents who do not receive appropriate audiological services may experience social isolation, cognitive decline and decreased mobility. PURPOSE: To examine the effectiveness of standard audiological testing procedures for nursing home residents and to discuss the purpose of adapting assessment procedures that can lead to more effective audiological assessments for this population. RESEARCH DESIGN: A retrospective chart analyses. A 33-item coding form was used to complete descriptive analysis of original audiological data and demographic data for 307 nursing home residents for a study to examine the effects of auditory stimulation on dementia-related behavior problems exhibited by nursing home residents through audiotape exposure to environmental sounds or soothing voice. RESULTS: Although 77% (n = 235) of the 307 residents were considered compliant for the testing process and 74% (n = 288) tolerated putting on headphones, audiological assessment using air conduction testing could be completed in both ears on 32% (n = 100) of the residents. In fact, only 5% (n = 16) of the 307 residents were able to complete a full traditional audiometric assessment protocol. CONCLUSIONS: Proper identification of hearing impairment through effective and appropriate audiological assessment is crucial for preserving and enhancing quality-of-life in nursing home residents. This study served as an introduction to the problem of using traditional behavioral testing for hearing assessment of nursing home residents. Much work needs to be done to establish best practices for audiometric assessment in this population. PMID- 19902703 TI - Longitudinal changes in real-ear to coupler difference measurements in infants. AB - BACKGROUND: The real-ear-to-coupler difference (RECD) measurement is a commonly used clinical procedure that quantifies the difference in sound pressure level between a 2 cc coupler and an individual's ear canal. The SPL levels in infant ears are highly variable and significantly higher than the SPL levels present in average adult ears, making the quantification of SPL levels in infant ears extremely important for threshold determination and fitting of amplification. It is unknown how much the RECD changes in an individual infant over time, whether that change is within test-retest reliability of the RECD measure, and whether RECD values are predictable from other outer and middle ear measures. PURPOSE: The purposes of this study were to examine longitudinal changes in RECD values in newborn infants to determine whether a significant change in RECD values takes place over a one-month period, how the change in RECD relates to test-retest variability of the measure, and whether RECD values are predictable from the infant's corrected age, or measures of static admittance and equivalent ear canal volume (EECV). STUDY SAMPLE: Fourteen infants (seven females, seven males) aged 7 to 25 days were recruited through community prenatal classes, physicians, hospital nurseries, and word of mouth. All infants had normal middle ear status. DATA COLLECTION AND ANALYSIS: Infants were tested on two separate visits, first when the infant was approximately two to three weeks old and then approximately one month later. Each visit lasted one to two hours, during which time otoacoustic emissions, 226 Hz and 1000 Hz probe-tone tympanometry, and two RECD measures were made for each ear. RESULTS: A multivariate analysis of variance revealed a significant change in RECD values over a one-month interval. Regression analyses revealed that final RECD values were partially predictable from age, initial RECD value, static admittance; and EECV. RECD test-retest variability was not large but larger than longitudinal RECD changes over a one month period. CONCLUSIONS: It may be unnecessary to remeasure an RECD to account for changes in ear canal acoustics for repeated assessments, no more than one month apart, when the same test transducer is used. The RECD should, however, be measured at one of these assessments because RECD values are not predictable from an infant's age or measures of static admittance and ear canal volume. PMID- 19902704 TI - The effect of visual and audiovisual competition on the auditory N1-P2 evoked potential. AB - BACKGROUND: Use of a competition stimuli (e.g., video) in clinical settings is a widespread practice, yet the effects of these stimuli on evoked potentials are not well understood. PURPOSE: The present investigation acquired the click evoked auditory N1-P2 in quiet and in two types of competition: during presentation of an unrelated visual stimulus or audiovisual stimulus. Responses were acquired in each of these conditions at two click stimulus levels (i.e., 35 and 65 dBnHL) to determine if the effect of competition was greater closer to threshold. An attempt was made to quantify effect robustness by examining within- and between session reliability. RESEARCH DESIGN: Repeated measures ANOVA. STUDY SAMPLE: 17 normal hearing female subjects. RESULTS: Findings indicated a significant effect of audiovisual competition on the amplitude of the N1-P2, and a borderline significant effect of visual competition on this index. The extent of visual competition was better conveyed when examined on a case-by-case basis, in which was revealed a subgroup of the sample that was negatively affected by visual competition. There was no interaction between competition and click stimulus level. CONCLUSIONS: Although competition stimuli can negatively affect the amplitude of the N1-P2, consideration should be given to subjects on a case-by case basis to warrant if removal of this stimulus is necessary. PMID- 19902706 TI - On the diagnosis of Auditory Processing Disorders. PMID- 19902705 TI - Effects of a cell phone conversation on cognitive processing performances. AB - BACKGROUND: The ability to apportion cognitive resources to process multiple visual and auditory stimuli is essential for human communication in competing conditions. PURPOSE: The purpose of the current research was to examine the effects of a cell phone conversation on a battery of cognitive tests, using both timing (RT) and accuracy (A') as dependent measures. RESEARCH DESIGN: A repeated measures ANOVA was conducted. STUDY SAMPLE: Forty-two college-age (mean 22 yr) adult females with normal hearing and cognitive function participated in the study. DATA COLLECTION AND ANALYSIS: In one condition (quiet), a standardized cognitive assessment battery was administered to participants in a quiet room. In the (cell phone) condition, subjects were formulating and responding to specific questions about their travel experiences during administration of the same cognitive assessment battery. The computer automatically records subject performance. Post-hoc pairwise comparisons were conducted using the Bonferroni approach. The alpha level was set at .05 for all data analysis. This method of analysis was repeated for each of the dependent measures, RT, and A'. RESULTS: The results revealed a consistent, significant effect on reaction time between the two conditions. The same analysis was also conducted to examine the effect of participation in a cell phone discussion on accuracy. As with RT, results revealed a consistent, significant affect on A' between the two conditions. CONCLUSIONS: Our study supports the notion that there are differential effects of auditory distracters across cognitive spheres. For simple automatic type visual cognitive tasks, the effect is minimal. However, as visual tasks increase in difficulty, the effect of the auditory distraction is magnified, particularly when the task requires extensive division of language resources. PMID- 19902707 TI - Precedent setting study: physicians are vital to Kansas City economy. PMID- 19902708 TI - Safeguarding the quality of health care in Kansas City. PMID- 19902709 TI - Canada's ObamaCare precedent. PMID- 19902710 TI - You're out of line, Mr. President AND you're wrong! PMID- 19902711 TI - Ethical concerns in the use of electronic medical records. AB - The use of electronic medical record systems raises important ethical concerns about patient privacy and confidentiality, medical errors, expectations of structured data entry by clinicians, documentation integrity, and provider patient interaction. Clinicians and health care organizations need to define best practices and policies in the use of EMR systems to improve quality and maintain clinician efficiency without compromising patient welfare and safety. PMID- 19902712 TI - Moving the field of gastroenterology & hepatology forward. PMID- 19902713 TI - Barrett's esophagus: a pre-cancerous condition approach to diagnosis and management. AB - Barrett's esophagus (BE) results from prolonged uncontrolled gastroesophageal reflux (GERD). Patients at risk for BE should be screened with upper endoscopy. Dysplasia is identified pathologically on endoscopic biopsy. The finding of low grade dysplasia indicates the need for more surveillance. High grade dysplasia warrants intervention with ablative techniques or surgery due to the extremely high rate of malignant transformation to esophageal adenocarcinoma. All patients should receive measures to control GERD (life-style modifications and acid suppression). PMID- 19902714 TI - Protecting the stomach from the "toxins" of NSAIDS and anti-platelet agents. AB - Non-steroidal antiinflammatory drugs (NSAIDs) and anti-platelet agents are some of the most commonly prescribed and utilized medications. There is overwhelming evidence of linking NSAIDs/anti-platelet agents to gastrointestinal toxicities, mouth to colon. In this article, we will review the effectiveness of interventions to prevent these GI-related toxicities. PMID- 19902715 TI - Celiac disease. AB - Celiac disease is a malabsorptive disorder resulting from intolerance to gluten and leads to systemic problems due to nutritional deficiencies. Enhanced diagnostic ability through serology has led to the understanding that it is more common than previously thought and often presents in atypical and subclinical forms. Clinicians must recognize the subtle presentations that may represent celiac disease, as well as at-risk populations, so that appropriate measures can be taken to diagnose and treat the disorder. PMID- 19902716 TI - Choices in colorectal cancer screening: a review of current screening modalities and recommendations. AB - Colorectal cancer (CRC) is the second leading cause of cancer death in the United States. In 2008, screening recommendations for CRC were updated. Successful reduction in CRC prevalence and deaths depends on a thorough understanding and correct implementation of these guidelines. This paper reviews the most recent CRC screening guidelines, in order to promote increased screening and effective adherence to these guidelines. PMID- 19902717 TI - Biologic therapy in Crohn's disease. AB - Crohn's disease is a complex disease process causing transmural inflammation of the gastrointestinal tract, which produces a constellation of symptoms including abdominal pain, frequent loose stools, fistula formation, and extraintestinal manifestations. Biologic therapy, including tumor necrosis factor alpha inhibitors, are a growing class of agents, which have demonstrated efficacy in the treatment of moderate to severe Crohn's disease. Their indications for use have grown, as has the understanding of their associated safety concerns. PMID- 19902718 TI - Chronic viral hepatitis. AB - Chronic liver disease is the tenth leading cause of death in the United States. The economic burden of liver disease is approximately 1% of the total national health care expenditure. Chronic hepatitis C is the most common cause of chronic liver disease in the United States, and chronic hepatitis B is the leading cause of liver disease worldwide. This article provides a brief overview of the evaluation and treatment of chronic hepatitis B and C infection. PMID- 19902719 TI - Hurthle cell carcinoma in a patient with Graves' Disease. AB - Hurthle cell, a rare form of thyroid carcinoma, has been reported only five times in patients with Grave's Disease. We present a case report and comprehensive review of the literature. Surgery should be considered in patients with both Graves' Disease and an expanding goiter. Initial high-dose radioiodine should be administered post-operatively in patients with Hurthle cell carcinoma. Serum thyroglobulin and 18-FDG total body PET imaging may be valuable in discovering residual or recurrent disease. PMID- 19902720 TI - Peripheral arterial thrombosis with presence of anti-centromere antibodies and polycythemia vera. AB - Digital gangrene due to arterial thrombosis can occur due to a variety of causes. There have been reports of patients who have presented with digital gangrene with no known predisposing factors, but with positive anti-centromere antibodies (ACA). Despite that they did not have features suggestive of disorders like Scleroderma or CREST syndrome. We report a patient with digital gangrene, who was asymptomatic before this episode and was found to have both ACA positivity and Polycythemia Vera. PMID- 19902721 TI - Nephrogenic systemic fibrosis: what internists need to know. AB - Nephrogenic systemic fibrosis (NSF) is an emerging entity that affects the skin and internal organs in patients with renal insufficiency and may end in death. More than 215 cases have been reported to the NSF registry at Yale University and more than 700 cases have been reported world-wide. There is a growing concern about gadolinium contrast media as being the major player in NSF development. Given the lack of established effective therapy, gadolinium avoidance along with careful consideration of the predisposing factors is the vital approach. Clinicians must be aware of the risks and benefits of gadolinium administration especially in patients with chronic kidney disease. PMID- 19902722 TI - Humor in intern retreats: scrubbing away the blues. AB - Humor in everyday practice has been shown to reduce work stress and increase efficiency. We aimed to study the effect of humor on interns for which we employed humor in one of the intern retreats and later asked the interns to rate the value of each retreat. Pre- and post- retreat mood states were also surveyed. The majority of interns recommended that humorous intern retreats be used in the coming years. A trend towards improvement in depression scores was also noticed. PMID- 19902723 TI - Value of preoperative templating for primary total knee arthroplasty. AB - The value of preoperative templating for primary total knee arthroplasty (TKA) has recently been questioned. In our clinical practice, preoperative templating is valuable for sizing components and achieving optimal coronal alignment. We preoperatively templated the size of femoral and tibial components, the distal femoral valgus cut, and the proximal tibial cut for 200 primary TKAs. Preoperative templating predicted the exact size of 165 (82.5%) femoral and 159 (79.5%) tibial components, with 194 (97%) femoral and 185 (92.5%) tibial components templated to within one size used. Postoperative tibiofemoral alignment was within +/- 3 degrees of the goal alignment in 189 (94.5%) knees. Postoperative femoral and tibial component coronal alignment was within +/- 3 degrees of the goal coronal alignment in 190 (95%) and 199 (99.5%) knees, respectively. The accuracy of component sizing in this study is greater than previously reported. The prevalence of alignment outliers in this study is less than previously reported for conventional techniques. PMID- 19902724 TI - Automatic and manual methodology for three-dimensional measurements of distal femoral gender differences and femoral component placement. AB - Our study aimed to introduce an automatic three-dimensional method for measuring the distal femur and identifying potential gender differences and the effects on femoral component placement in total knee arthroplasty. Three hundred forty-two femora were scanned with computed tomography. Automatic and manual bone resection and component placement were compared using a virtual resection tool. For standard component use, 77.3% of the femora were male and 23.1% were female. For gender-specific component use, 91% were female and 7.3% were male. Surgeon errors in both component translation and rotation existed but were minimal. From these results, gender alone did not dictate component use in primary total knee arthroplasty. The restoration of femoral condylar profile in 3 dimensions can be obtained by accurately measuring patient distal femoral anatomy and the appropriate femoral component design selection. Additional bone cuts, soft-tissue maneuvers, and adverse outcomes in fitting the patient to the femoral component may be avoided. PMID- 19902725 TI - Comparison of total knee arthroplasty in stiff knees and knees with good preoperative range of motion. AB - This study compared the results of total knee arthroplasty (TKA) in patients with stiff knees and patients with good preoperative range of motion. Results following TKA were compared in 30 patients (52 knees) with severely degenerated knees with preoperative range of motion < 50 degrees (group 1) and 30 patients (50 knees) who were matched for age, gender, and diagnosis with preoperative range of motion > 90 degrees (group 2). Results were evaluated for range of motion and Knee Society score, as well as the need for extensile approaches, soft tissue releases, additional bone cuts, and constrained prostheses. Radiographic results were evaluated for implant positioning, patellar height, and alteration in joint line. Findings indicated that although TKA in stiff knees can be successful, the results are inferior to those obtained in standard TKA. Careful preoperative planning and meticulous surgical technique are essential for good results. PMID- 19902726 TI - Patellofemoral kinematics after limited resurfacing of the trochlea. AB - Patellofemoral kinematics after a limited resurfacing of the trochlea was investigated. Patellofemoral contact pressure, area, and force were measured for intact state; after creation of a 20-mm full-thickness trochlear defect; and after trochlear resurfacing implant (Arthrosurface) in serial flexion positions (45 degrees, 60 degrees, 75 degrees). In the defect state, edge loading and peak contact forces were highest at the periphery. The chondral defect increased peak contact force (13 to 18 N, P < .01) and peak contact pressure (23 to 31 kg/cm2, P < .02) compared with the intact state. Peak contact pressure and force were restored to 90% (P < .01) and 88% (P < .01). Implantation of the device restored contact area to 85% of the intact state. Limited resurfacing of the trochlea restores contact area, peak pressure, and peak force to the intact state. These findings highlight the potential clinical use of limited patellofemoral resurfacing in trochlear chondral defects. PMID- 19902727 TI - Evaluation of the functional effects of anterior cruciate ligament bundles: a cadaveric experiment. AB - The influence of isolated deficiency of the anteromedial or posterolateral bundles of the anterior cruciate ligament (ACL) on knee kinematics has not been fully investigated. Thirty-two cadaveric knees were studied. The fibers of the anteromedial and posterolateral bundles were resected arthroscopically in alternating order in right and left knees. Before and after each arthroscopic cut, laxity tests were performed. Positive results on anterior drawer tests were specific only to the anteromedial bundle-cut knees. Pivot shift tests were positive only in the posterolateral bundle-cut knees. In addition, anterior tibial translation was measured with KT-1000 in response to different external loading conditions. Anterior translation measured with KT-1000 at 67 N and 89 N draw forces at 20 degrees and 40 degrees of flexion may be used in evaluating the integration of each bundle of ACL, both separately and as a whole. PMID- 19902728 TI - Assisted reproduction of the anterior cruciate ligament. AB - This article describes a technique for anterior cruciate ligament (ACL) repair using a synthetic braided ligament made of 75% degradable polyglycolic acid filaments and 25% nondegradable, 6.5-mm Dacron thread, wrapped in a free synovial graft. In a prospective study of 70 consecutive patients, this technique (experimental group) was compared to a standard bone-patellar tendon-bone autograft technique (autograft group). All of the patients improved after surgery. There were no surgical complications, and all of the patients returned to full activity without pain or instability. After mean follow-up of 12 years (range, 8-19 years), patients in the experimental group functioned well and statistically better than patients in the autograft group using International Knee Documentation Committee, Knee Injury Osteoarthritis Outcome Score, and Lysholm evaluation measurement tools. These results suggest a partially biodegradable replacement ligament prosthesis protected by a synovial wrap can result in a functionally stable knee. PMID- 19902729 TI - Knee articular cartilage injuries in the National Football League: epidemiology and treatment approach by team physicians. AB - This study describes the epidemiology of knee articular cartilage injuries in the National Football League (NFL) and the typical treatment approach by NFL team physicians. All knee articular cartilage injuries in the league database from 1992 to 2006 were reviewed, and team physicians were surveyed about their treatment approach to these injuries. A total of 118 cases were identified, for an average of 8 per year, with a higher injury rate during games compared with practice. Approximately half of the injuries occurred in linemen, and the most common site of injury was the femoral condyle. Forty-six percent of these injuries were treated surgically, with slower return to play after surgery (124 versus 36 days, P < .01). Among team physicians, microfracture was the most popular treatment approach (43%), followed by debridement (31%) and nonoperative management (13%). More research is needed to compare long-term outcomes based on lesion size in these athletes. PMID- 19902730 TI - Posterior medial meniscus detachment: a unique type of medial meniscal tear. AB - Patients with posterior medial meniscal detachment, as determined at knee arthroscopy, were evaluated retrospectively. Mean follow-up was 5.3 years for 8 men and 20 women (30 knees; mean age, 57 years). Most patients had acute onset of pain with a minor specific incident. Seventeen patients were obese, 9 were overweight, and 2 were normal. Eleven of 22 magnetic resonance imaging evaluations detected a tear at the site of the posterior medial meniscus root. Nine of 16 bone scan evaluations showed moderate uptake medially. Arthroscopic treatment included partial medial meniscectomy or meniscal repair. Twelve knees (40%) showed significant progression of arthritis. Of the 7 patients with severe arthritic knees, 5 have subsequently undergone total knee arthroplasty, 1 is considering total knee arthroplasty, and the other has minimal symptoms. Patients should be counseled about the clinical course of posterior medial meniscus detachment and its potential for progressive arthritis in the joint. PMID- 19902731 TI - Meniscus status at anterior cruciate ligament reconstruction associated with radiographic signs of osteoarthritis at 5- to 10-year follow-up: a systematic review. AB - The development of premature osteoarthritis after anterior cruciate ligament (ACL) reconstruction is a significant cause of morbidity in young, active individuals. Meniscal injuries are frequently noted at the time of reconstruction, and the critical role of an intact meniscus in the prevention of osteoarthritis has been well documented. The purpose of this review is to evaluate the effects of meniscal status at ACL reconstruction on the subsequent development of osteoarthritis. A systematic review of the literature identified 11 studies with > or = 2 years of follow-up that compared patients' radiographic outcomes based on meniscus status at the time of ACL reconstruction. Patients undergoing partial meniscectomy at the time of ACL reconstruction were significantly more likely to develop radiographic evidence of osteoarthritis than those with normal menisci. Meniscal repair resulted in inconsistent findings. Virtually all patients who underwent complete meniscectomy at the time of ACL reconstruction had radiographic evidence of osteoarthritis at follow-up. PMID- 19902732 TI - Patellar impingement by synovial cord following anterior cruciate ligament reconstruction. AB - We found a cordlike structure (synovial cord) in the patellofemoral joint causing patellar impingement in a 45-year-old man after anterior cruciate ligament (ACL) reconstruction. Arthroscopic examination revealed that a cylindrical cordlike soft tissue with a smooth surface extended between the medial and lateral capsule of the patellofemoral joint. The tissue histologically consisted of dense collagen fibers in parallel bundles covered with thin layers of synovial tissue; in addition, there was a proliferation of small vessel patches surrounded by hemosiderin-laden macrophages, which indicated bleeding episodes due to impingement in the patellofemoral joint. To our knowledge, this is the first case report of a synovial cord in the patellofemoral joint causing patellar impingement following ACL reconstruction. PMID- 19902733 TI - Extension loss secondary to femoral-sided inverted cyclops lesion after anterior cruciate ligament reconstruction. AB - Loss of terminal knee extension after anterior cruciate ligament (ACL) reconstruction is problematic. Formation of a fibrous nodule, known as a cyclops lesion, at the site of the ACL graft exiting from the tibial tunnel can impinge at the roof of the intercondylar notch and block terminal knee extension. This article presents a case of an inverted cyclops lesion originating from the femoral tunnel site of the ACL graft, representing a variant of the traditional cyclops lesion. The lesion was identified by clinical examination including prone heel-height difference assessment in combination with magnetic resonance imaging arthrography. The lesion was treated successfully by arthroscopic resection, with rapid resolution of the patient's painful, symptomatic extension loss. PMID- 19902734 TI - Supracondylar femoral fracture after anterior cruciate ligament reconstruction with transfemoral fixation. AB - Femoral fractures following anterior cruciate ligament (ACL) reconstruction are rare. These injuries often are related to increased stresses at the femoral tunnel. Hamstring tendon autograft is often used for ACL reconstruction, and transfemoral fixation is one of many graft fixation techniques with few reported complications. This article reports an atraumatic transverse supracondylar femoral fracture occurring through the transfemoral fixation tract following hamstring ACL reconstruction in a 38-year-old woman. PMID- 19902735 TI - Irreducible posterolateral knee dislocation with entrapment of the adductor magnus tendon and medial skin dimpling. AB - This case report describes an irreducible posterolateral knee dislocation with a bone avulsion of the medial epicondyle attached to the medial collateral ligament and adductor magnus tendon entrapped within the joint space. PMID- 19902736 TI - Partial femoral avulsion of the posterior cruciate ligament presenting as an osteochondral defect. AB - A 22-year-old soldier presented with mild anterior knee pain. Routine knee radiographs and magnetic resonance imaging revealed bony fragments that suggested an osteochondral defect in the medial femoral condyle. Arthroscopy revealed the fragment originated from a posterior cruciate ligament (PCL) avulsion and a well located intercondylar notch with a connection to the PCL fiber. An initial attempt was made to fix the fragment at the original avulsion site. However, the fragment was deformed, and the avulsion site was incongruent after reduction. The fragment was debrided accordingly, and the defect site was filled with an osteochondral graft from a nonweight-bearing portion of the lateral femoral condyle. The patient's postoperative course was uneventful, and he was able to participate in military training without discomfort. PMID- 19902737 TI - Donor-site giant cell reaction following backfill with synthetic bone material during osteochondral plug transfer. AB - Osteochondral defects are common in younger, active patients. Multiple strategies have been used to treat these lesions, including microfracture and osteochondral plug transfer. We describe a patient experiencing chronic knee pain and a full thickness cartilage defect on the lateral femoral condyle. After failing conservative management and microfracture surgery, the patient underwent osteochondral autograft plug transfer, with backfilling of the donor sites using synthetic bone graft substitute. Initial recovery was uncomplicated until the patient experienced pain following a twist of the knee. Magnetic resonance imaging for the subsequent knee injury revealed poor healing at the donor sites. The donor sites were debrided, and specimens revealed a foreign body giant cell reaction. Donor-site morbidity is of primary concern during osteochondral plug transfer; however, insufficient data exist to support the use of synthetic bone graft material. Our results indicate that off-label use of synthetic bone graft substitute during a primary procedure requires further investigation. PMID- 19902738 TI - Fractured polyethylene tibial post in a posterior-stabilized knee prosthesis presenting as a floating palpable mass. AB - Complications associated with posterior-stabilized total knee arthroplasty (TKA) include patellar clunk syndrome, posterior subluxation of the tibial component, tibial post wear, and tibial post failure. Recently, an increasing number of reports have been issued on polyethylene tibial post failure in posterior stabilized TKA. In most of these cases, tibial post failure in a well-functioning posterior-stabilized TKA was suspected based on clinical symptoms, such as a sudden onset of pain, effusion, patellar clunking, knee instability, and prosthesis dislocation. However, a floating, palpable, hard, mass-like symptom in the knee joint has not been previously reported. The authors report a case of a fractured polyethylene tibial post in a posterior-stabilized TKA that manifested as a palpable mass in the suprapatellar pouch. PMID- 19902739 TI - Synovial sarcoma presenting as iliotibial band friction syndrome. AB - Iliotibial band friction syndrome is a common entity that is often quickly diagnosed in orthopedic clinics. However, synovial sarcoma is an elusive clinical entity that appears around many joints with variable presentations. This case report is an example of a patient with a classic presentation of iliotibial band friction syndrome that was diagnosed as a synovial sarcoma on further investigation. PMID- 19902740 TI - Functional reconstruction of a chronically ruptured extensor apparatus after patellectomy. AB - Rupture of the knee extensor mechanism after previous patellectomy is rare. A treatment protocol therefore is not available. This article presents a new and easy technique using a gracilis-semitendinosus tendon autograft for successful reconstruction of a ruptured knee extensor mechanism after previous patellectomy. PMID- 19902741 TI - Semitendinosus allograft reconstruction of chronic biceps femoris rupture at the knee. AB - Surgical treatment of distal hamstring ruptures at the knee is rare and has been reported infrequently in the literature. This article describes a 22-year-old former collegiate football player who had his career cut short secondary to a chronic distal rupture of the biceps femoris at the knee. Reconstruction of the distal biceps femoris tendon with semitendinosus allograft resulted in an excellent clinical outcome and allowed the patient to return to a high level of physical activity. This case presents a unique cause of knee pain and weakness in a young, active patient and a corresponding effective surgical treatment for such an injury. PMID- 19902742 TI - [Observing problems in surgical treatment of lumbar spinal stenosis]. PMID- 19902743 TI - [Effect of pulsed electromagnetic field on the changes of osteoclasts in ovariectomized rats bone marrow culture in vitro]. AB - OBJECTIVE: To investigate the effect of pulsed electromagnetic fields (PEMs) on inducing osteoclastic like cell (OLC) formation changes and apoptosis in ovariectomized (OVX) rats bone marrow culture in vitro. METHODS: Thirty healthy three-month-old female Wistar rats were either sham-operated (Sham) or ovariectomized (OVX) and randomly divided into three groups: group A (OVX + PEMs, 18 rats), group B (OVX, 6 rats) and group C (Sham, 6 rats); group A was again randomly divided into three groups: A1, A2, A3. The frequencies adopted were 1.5, 2, 75 Hz and 30 minutes for once a day. All rats were fed with normal diet for 3 months, then the bone marrow of all rats were cultured, 2 days later, group A cells (including group A1, A2, A3) were collected and exposed to different frequencies PEMs for 2 weeks (30 min/day). In order to observe the changes of osteoclasts and count their numbers, cells were taken for Wright Giemsa staining, tartrate-resistance acid phosphatase (TRAP) staining and Hoechst 33258 staining. RESULTS: TRAP staining results indicated the number of OLC in group C was the least, then was group A2, A3, A1, B. The number of OLC in group B was remarkably increased (P < 0.01; vs group C, A2). The number of OLC in group B was significantly increased (P < 0.05; vs group A1, A3). Hoechst 33258 staining results indicated the number of apoptosis of OLC in group C was more than other groups, which of group C, A2 was significantly increased (P < 0.05; vs group B). CONCLUSION: PEMs had decreased the formation of OLC and increased the number of apoptosis of OLC in ovariectomized (OVX) rats bone marrow culture in vitro, the effects of 2 Hz was the best. PEMs would be a new way of osteoporosis therapy. PMID- 19902744 TI - [Imageology change of the intervertebral foramen degenerative intervertebral disc in different degrees and its clinical]. AB - OBJECTIVE: To study the imageology change of the intervertebral foramen degenerative intervertebral disc in different degrees and explore its clinical significance. METHODS: The imageology data (MRI and CT) of 37 patients with degenerative disc disease of L4,5 (male 23, female 14, age from 28 to 62 years with an average of 41.6 years)were investigated. The patients were divided into three groups depending on the mean signal intensity rate of degenerative disc and cerebrospinal fluid:light degenerative group (group A) of 11 cases, intermediate degenerative group (group B) of 13 cases, and severe degenerative group (group C) of 13 cases. The extreme altitude, maximum width and areas of the intervertebral foramen were measured from the CT 1.25 mm scan reconstitution. The changes of the intervertebral foramen were analyzed. RESULTS: (1) 1. The extreme altitude and areas of the intervertebral foramen gradually diminished among the light degenerative group, intermediate degenerative group and severe degenerative group, there was no significant deviation between intermediate degenerative group and the light degenerative group (P > 0.05), there was statistical significance between severe degenerative group and intermediate degenerative group (P < 0.05), there was statistical significance between severe degenerative group and light degenerative group (P < 0.01). (2) There was no statistical significance of the maximum width of intervertebral foramen among three groups (P > 0.05). CONCLUSION: The extreme altitude and areas of the intervertebral foramen gradually diminished when the disc are differently degenerative. But there was not significant correlation to width of the intervertebral foramen; the dimin height and area of intervertebral foramen should result in root compression. PMID- 19902745 TI - [Clinical control study in treating degenerative lumbar instability with single or double carbon fiber cages]. AB - OBJECTIVE: To compare the clinical and radiologic effect in treating degenerative lumbar instability with single or double carbon fiber cages. METHODS: From November 2005 to October 2006, 97 patients of degenerative lumbar instability were followed up more than two years. All cases underwent pedicle screw fixation. Meanwhile, 41 of them, named group A, were applied with single carbon fiber cage, with 23 males and 18 females, aged for 35-70 years, including 38 cases with single vertebral instability and 3 cases with existing L4.5 and L5S1 intervertebral instability. The other 56 cases, named group B, were treated with two carbon fiber cages, with 32 males and 24 females, aged for 33-72 years, including 53 cases with single vertebral instability and 3 cases existing L4.5 and L5S1 intervertebral instability. The clinical effect, intervertebral fusion ratio and lost intervertebral height were compared between two groups. RESULTS: All patients were followed up for 24-35 months, with an average of 28 months. Clinical effects (including symptom, sign, daily action and bladder function) were evaluated according to JOA 29 score. In group A: the JOA score improve from 10.11 +/- 2.40 preoperative to 24.88 +/- 2.30 at final follow-up, 25 cases obtained excellent results, 12 good, 4 fair; 39 cases obtained fusion with the fusion ratio of 95.1% (39/41). In group B: the JOA score from 9.62 +/- 2.60 preoperative to 25.19 +/- 2.40 at final follow-up, 37 cases obtained excellent results, 13 good, 6 fair; 53 cases obtained fusion with the fusion ratio of 94.6% (53/56). In the 7th day after operation, the average intervertebral height in group A was (11.2 +/- 1.2) mm,and that of group B was (11.3 +/- 1.4) mm. Two years later, the average intervertebral height in group A was (11.0 +/- 1.4) mm, while that of group B was (11.1 +/- 1.4) mm. There was no significant difference between two groups in clinical effect, intervertebral fusion ratio and lost intervertebral height (P > 0.05). CONCLUSION: Pedicle screw fixation and single or double carbon fiber cages in treating degenerative lumbar instability both can obtain satisfactory curative effect. Single carbon fiber cage has advantages such as minimally trauma, simply operation, retentively posterior column structure, cheaply price than double carbon fiber cages. PMID- 19902746 TI - [Surgical treatment for lumbar spinal stenosis by different methods]. AB - OBJECTIVE: To discuss the relationship between decompression methods of lumbar spinal stenosis and outcomes. METHODS: From September 1996 to March 2007, 68 patients with degenerative lumbar spinal stenosis were investigated retrospectively. There were 40 males and 28 females with the mean age of 52.8 years (ranging from 32 to 78 years old). And the average history was 51 months. According to Hansraj classification of lumbar spinal stenosis and levels response to symptom,the classic lumbar spinal stenosis were treated with complete laminectomy and foramintomy (18 cases in group A), hemilaminectomy and foreminotomy (11 cases in group B), complete fenestration and foraminotomy (17 cases in group C); and other patients with complicated lumbar spinal stenosis (22 cases in group D) were treated with decompression and intervertebral fusion and internal fixation. The clinical results of all patients were analyzed according to JOA score (15 scoring method). RESULTS: All patients were followed up with an average of 64 months (ranging from 8 months to 11 years). At final follow-up,the improvement rate of JOA score in the group A,B,C,D was respectively (51.2 +/- 26.6)%, (60.7 +/- 21.1)%, (59.3 +/- 23.1)% and (59.1 +/- 22.7)%. These data were significantly difference than that of preoperative (P < 0.001). CONCLUSION: CT and MRI results combining with clinical symptom and sign is the key to determine decompressive extent; lumbar stability is the key to determine fixation and intervertebral fusion in treating lumbar spinal stenosis. PMID- 19902747 TI - [Selective fenestration lamina of cervical vertebral lamina to excise ligamentum flavum for treating hypertrophic ligamentum flavum]. AB - OBJECTIVE: To explore operative method and clinical effect of selective fenestration of cervical vertebral lamina to excise ligamentum flavum for treating hypertrophic ligamentum flavum (HLF). METHODS: From March 1998 to May 2007, 21 patients (14 males and 7 females) with HLF were reviewed retrospectirely, whose age was from 37 to 76 years with an average of 52.2 years. The history of HLF was from 5 h to 25 years with an average of 3 years and 10 months. Reason of onset: 4 cases were no inducement, 10 cases were slow moving,and 7 cases caused by injury. The operative effect was assessed according to JOA scores and radiologic results. RESULTS: All patients were followed up with an average of 2.5 years(ranging from 3 months to 7.5 years). Radiologic results: sagittal diameter of vertebral canal and trans.sec.area increased and spinal cord were well than preoperative status. The majority of cases got stronger in the walking, improvement in the fingers mobility and holding power and using chopsticks, abatement in the limbs numb and thoracic waist zonesthesia, amelioration in the urination and defecation functional disturbance than preoperative symptom. The JOA scoring was 5-16 scores with an average of 11.48 +/ 2.94 before operation,and 7-17 scores with an average of 13.81 +/- 2.98 after operation. That results had statistical significance between before and after operation (t = 3.51, P < 0.05). There were 13 cases excellent (improvement rate more than 75%), 5 cases good (improvement rate in 50%-74%), 2 cases fair (improvement rate in 25%-49%), 1 case poor (improvement rate less than or equal to 24%) with an average improvement rate of 85.2%. CONCLUSION: It is simple in the operative procedure and tiny in trauma in treatment for hypertrophic ligamentum flavum with selective fenestration of cervical vertebral lamina to excise ligamentum flavum. The method can obtain effective decompression for vertebral canal but choice of indication must be reasonable. PMID- 19902748 TI - [Clinical analysis in treating lumbar intervertebral disc herniation with nucleus pulposus resection through small incision and lamina fenestration]. AB - OBJECTIVE: To analyze effect of treatment of lumbar intervertebral disc herniation with nucleus pulposus resection through small incision and lamina fenestration,and to explore operative advantage and mattars needing attention. METHODS: Ninety-six patients with lumbar intervertebral disc herniation, including 48 males and 48 females with an average age of 46.4 years (ranging for 16-75 years) and an average course of 5 years (ranging from 1 month to 30 years), were treated with nucleus pulposus resection through small incision and lamina fenestration. The clinical effects were analyzed according to JOA scoring criteria of lower back pain. RESULTS: All patients were followed up from 6 months to 3.5 years with an average of 1.2 years. According to JOA scoring criteria, 34 cases obtained excellent result, 55 good, fair 7. The rate of excellent and good was 92.71% and the mean improvement rate was 54.53%. CONCLUSION: Nucleus pulposus resection through small incision and lamina fenestration can obtain satisfactory outcome in treating lumbar intertebral disc herniation. The incidence of lumbar instability and postoperative complications were low. It is one of the better method for the treatment of lumbar intertebral disc herniation. PMID- 19902749 TI - [Imaging diagnosis of lumbar spinal stenosis]. AB - OBJECTIVE: To investigate value of X-ray, CT and MRI for the diagnosis of lumbar spinal stenosis. METHODS: The data of 130 patients with clinical diagnosis and typical imaging signs of lumbar spinal stenosis were analyzed. The present study included 83 males and 47 females with an average age of 43.5 years (range from 27 to 75 years). CT examination was performed in all patients, routine X-ray examination in 23 patients and routine MRI in 57 patients. RESULTS: The lumbar spinal stenosis showed at L(3,4) plane in 25 patients, L(4,5) in 48 patients and L5S1 in 57 patients. CT showed hyperostosis of lumbar posterior marginal, vertebral lamina, inferior articular process in 46 patients, hypertrophy of superior and inferior articular processsus in 7 patients, calcification or ossification of ligamentum flavum in 13 patients, vertebral body spondylolisthesis in 5 patients, lateral recess stenosis in 24 patients, and intervertebral foramen stenosis in 35 patients. MRI showed intervertebral disk hernia with disc associated with ligamentum flavum hypertrophy in 23 patients, ligamentum flavum symmetrical hypertrophy in 18 patients,extensive multi segmental ligamentum flavum hypertrophy in 9 patients,and local ligamentum flavum hypertrophy in 7 patients. CONCLUSION: The main cause of secondary lumbar spinal stenosis is degeneration. Traditional X-ray examination has great limitations in diagnosis of lumbar spinal stenosis. CT and MRI have advantages of multi directional imaging and the high resolution. CT can show well ligament calcification and ossification and other bone change which are showed not well on MRI, so CT is recommended to lumbar spinal stenosis. PMID- 19902750 TI - [Treatment of Zone II flexor tendon injury]. PMID- 19902751 TI - [Micro-decompression procedure for lumbar spinal stenosis]. AB - OBJECTIVE: To evaluate clinical result of the micro-decompression procedure for lumbar spinal stenosis. METHODS: From September 2001 to May 2006,87 patients (male 60, female 27) with lumbar spinal stenosis underwent micro-decompression. The age of patients were from 43 to 80 years with an average of 51 years. Among them,2 cases with spinal stenosis occured in L(3,4), 47 in L(4,5), 38 in L5S1. RESULTS: All patients were followed up for 18-48 months with an average of 26 months. The results were excellent in 52 cases, good in 28, poor in 7, according to Macnab of back leg pain standard. The rate of excellent and good was 92%. CONCLUSION: Operative treatment for lumbar spinal stenosis is focused at the areas causing symptomatic neural root compression rather than prophylactic decompression at areas of nonsymptomatic disease. The micro-decompression procedures are more likely to be well tolerated by older patients. PMID- 19902752 TI - [Chiropractic adjustment combined with Buyang Huanwu decoction for the treatment of lumbar lateral recess stenosis]. PMID- 19902753 TI - [Treatment of lumbar intervertebral disc herniation accompanying with lumbar instability with internal fixation and spinal fusion]. AB - OBJECTIVE: To explore operative effect of lumbar intervertebral disc herniation accompanying with lumbar instability. METHODS: Form June 2000 to June 2006, 46 patients of lumbar intervertebral disc herniation accompanying with lumbar instability were treated with decompression through posterior approach, diskectomy, spinal fusion and vertebral pedicle internal fixation. Including 33 males and 13 females,the age was from 37 to 68 years with an average of 48 years. The course of disease was from 4 months to 20 years with an average of 3.5 years. There were simple segment in 21 cases, double segments in 22 cases, three segments in 3 cases. RESULTS: All patients were followed up for 12-45 months with an average of 25 months. All cases got solid fusion and clinical symptom improved obviously. According to clinical standard to evaluation, 32 cases obtained excellent result, 8 good, 6 fair. The rate of excellent and good was 86.9%. CONCLUSION: Diskectomy, spinal fusion and internal fixation can obtain satisfactory clinical effect for lumbar intervertebral disc herniation accompanying with lumbar instability. PMID- 19902754 TI - [Microscope-assisted minimally invasive decompression for degenerative lower lumbar spinal stenosis]. AB - OBJECTIVE: To evaluate clinical results of microscope-assisted minimally invasive decompression for degenerative lower lumbar spinal stenosis. METHODS: From May 2007 to November 2008, 26 patients with degenerative lower lumbar spinal stenosis were treated with microscope-assisted minimally invasive decompression. Including 9 males and 17 females, the mean age were 53.7 years (range in 47-75 years). The course of disease was from 2 to 8 years with an average of 3.6 years. Single segment symptom had 20 cases and double segments had 6 cases. The clinical effects were evaluated and improvement rate was calculated according to JOA scoring before and after operation. RESULTS: The operative time in single segment was 60-90 minutes with an average of 75 minutes and in double segments was 80-180 minutes with an average of 95 minutes. The operative bleeding in single segment was 50-120 ml with an average of 85 ml and in double segments was 60-150 ml with an average of 100 ml. All the patients were followed up with an average of 8.6 months (range in 6-24 months). The clinical effects were evaluated according to JOA scoring,excellent in 18 cases,good in 7,poor in 1,the rate of excellent and good was 96% and improvement rate was 86%. The mean JOA scoring was respectively 3.92 +/- 0.83 and 12.67 +/- 1.92 before and after operation. There was significantly statistic difference between before and after operation (P < 0.01). CONCLUSION: Microscope-assisted minimally invasive decompression for degenerative lower lumbar spinal stenosis can obtain satisfactory therapeutic effect,which had advantages of short operative time, less trauma, subtle operation, fast restoration, but it is inapplicable for the patients with obviously ossification or calcification of vertebral body, severe vertebral canal adherence. PMID- 19902755 TI - [Evaluation on analgesic effect of electroacupuncture combined with acupoint injection in treating lumbar intervertebral disc herniation]. AB - OBJECTIVE: To explore the analgesic effect of electroacupuncture (EA) combined with acupoint-injection in treating lumbar intervertebral disc herniation. METHODS: Sixty patients with lumbar intervertebral disc herniation were divided randomly into treatment group (30 cases, group A) and control group (30 cases, group B) according to random digits table. Involved 31 males and. 29 females in the study,the age was from 35 to 62 years with an average of 48.3 years. The patients in group A were treated with EA (once a day, a total of 10 times) and acupoint-injection (once every two days, a total of 5 times) at L4 Jiaji (EX-B2), L5 Jiaji (EX-B2), Zhibian (BL54), Huantiao (GB30), Yanglingquan (GB34), Weizhong (BLA0) and Kunlun (BL60); and the patients in group B were treated with single EA (Once a day, a total of 10 times). After 10 days of treatment, the visual analogue scale (VAS) was adopted to examine and compare the main pain score, and the clinical effects were compared between two groups from the symptoms and signs in accordance with efficacy criteria. RESULTS: The paregoric effect of group A was better than that of group B, there was significant differences in the main pain score between two groups (t = 7.18, P < 0.01). The clinical effect of group A was better than that of group B,there was significant differences between two groups (u = 2.91, P < 0.01). CONCLUSION: EA combined with acupoint-injection is a convenient and highly effective therapy for lumbar intervertebral disc herniation, which can effectively relieve the pain of the patients, should be to promote clinical use. PMID- 19902756 TI - [Manipulative reduction and closed thread a Kirschner wire for the treatment of comminuted fracture of distal radius in middle-aged and senile patients]. PMID- 19902757 TI - [Selective cervical dorsal root cutting off part of the vertebral lateral mass fixation combined with exercise therapy for treating spastic cerebral paralysis of the upper limbs caused by cerebral palsy]. AB - OBJECTIVE: To explore the feasibility and the result for the surgical treatment of spastic cerebral paralysis of the upper limbs in patients who underwent the selective cervical dorsal root cutting off part of the vertebral lateral mass fixation combined with exercise therapy. METHODS: From March 2004 to April 2008, 27 patients included 19 boys and 8 girls, aging 13-21 years with an average of 15 years underwent selective cervical dorsal root cutting off part of the vertebral lateral mass fixation with exercise therapy. The AXIS 8 holes titanium plate was inserted into the lateral mass of spinous process through guidance of the nerve stimulator, choosed fasciculus of low-threshold nerve dorsal root and cut off its 1.5 cm. After two weeks, training exercise therapy was done in patients. Training will include lying position, turning body, sitting position, crawling, kneeling and standing position, walking and so on. Spastic Bobath inhibiting abnormal pattern was done in the whole process of training. The muscular tension, motor function (GMFM), functional independence (WeeFIM) were observed after treatment. RESULTS: All patients were followed up from 4 to 16 months with an average of 6 months. Muscular tension score were respectively 3.30 +/- 0.47 and 1.25 +/- 0.44 before and after treatment;GMFM score were respectively 107.82 +/- 55.17 and 131.28 +/- 46.45; WeeFIM score were respectively 57.61 +/- 25.51 and 87.91 +/- 22.39. There was significant improvement before and after treatment (P < 0.01). CONCLUSION: Selective cervical dorsal root cutting off part of the vertebral lateral mass fixation combined with exercise therapy was used to treat spastic cerebral paralysis of the upper limbs is safe and effective method, which can decrease muscular tension and improve motor function, which deserves more wide use. PMID- 19902758 TI - [Diagnosis and treatment of popliteal vascular injury associated with knee joint trauma]. PMID- 19902759 TI - [Comparison of two approaches for the treatment of supracondylar fractures in children by K-wires]. AB - OBJECTIVE: To explore the clinical effect between two approaches for the treatment of supracondylar fractures of humerus by K-wires in children. METHODS: From Jan. 2004 to Dec. 2006,117 children with supracondylar fractures of humerus were divided into two groups according to different methods of the K-wires fixation involving two K-wires group and three K-wires group. In two K-wires groups,there were 45 children including 31 boys and 14 girls with an average age of 5.6 years ranging from 1 to 11 years; according to Garland classification, 19 cases were type II, 26 cases were type III. In three K-wires groups, there were 72 children including 47 boys and 25 girls with an average age of 6.8 years ranging from 2 to 12 years; according to Garland classification, 22 cases were type II, 50 cases were type II. The movement and carrying angle of elbow joint were measured for all these patients. According to Flynn criteria for supracondylar fracture the results of two approaches with K-wires were compared. RESULTS: All patients were followed-up from 2 to 24 months (means 15.4 months). In two K-wires group, the results of Flynn were excellent in 27 cases, good in 12 cases, fair in 4 cases and poor in 2 cases, 41 cases gained bony healing, however, fail of fixation arosed in 4 cases and cubitus varus arosed in 2 cases. In three K-wires group, the results of Flynn were excellent in 60 cases, good in 11 cases and poor in 1 case, all cases of three K-wires group achieved bony healing after 6 weeks of operation, the function of joints recovered through exercise and cubitus varus arosed in one case after operation. CONCLUSION: The technique of fixation with K-wires is a stable and reliable methods for unstable supracondylar fracture but medial-lateral three-pin fixation is better than one with two pins. PMID- 19902760 TI - [Influence of operative time on effect of the Pilon fracture]. AB - OBJECTIVE: To evaluate the influence of operative time on treating high-energy tibial Pilon fracture. METHODS: From June 2003 to May 2008, Sixty-five patients with tibial Pilon fracture were followed up. There were 42 males and 23 females in the study. The age was from 15 to 62 years with an average of (37.53 +/- 6.32) years. The patients were divided into two groups (group A and B) according to operative time. In the group A, 30 cases were treated with open reduction internal fixation (ORIF) at 3-7 days after injury. In the group B, 35 cases were treated with step-by-step postphoned ORIF at 7-24 days after injury (the swell of soft tissue extincted). The clinical effects were evaluated between two groups according to Bourne standard. RESULTS: All patients were followed up from 7 to 24 months with an average of 16.23 months. There was significant difference on the early complications between two groups. In group A, 3 cases of infection, 4 cases of edge splitting, 1 case of plate explosing and 1 case of osteomyelitis. In group B, only 2 cases of surface infection. The time of fracture healing in group A and B was respectively (18.3 +/- 3.2) weeks and (15.7 +/- 2.5) weeks. There was significant difference between two groups (P < 0.05). According to the Bourne Pilon fracture healing standard,in group A, 12 cases got excellent result, 10 good, 8 poor, the rate of excellent and good was 73.3%; in group B, 21 cases got excellent result, 8 good, 6 poor, the rate of excellent and good was 82.9%. There was significant difference between two groups (P < 0.05). CONCLUSION: The choice of operative time is the key to treat high-energy tibial Pilon fracture. Step-by step postphoned ORIF is an effective method for high-energy tibial Pilon fracture. PMID- 19902762 TI - [Manipulative reduction and small splint fixation for the treatmemt of closed tibial fractures]. PMID- 19902761 TI - [Effect of the mixed liquor of danshen and magnesium sulfate injection on IgG and IgM in serum of rabbits with lumbar intervertebral disc herniation]. AB - OBJECTIVE: To explore the effect of the mixed liquor of danshen and magnesium sulfate injection on inflammatory reaction caused by autoimmune response of rabbits with lumbar intervertebral disc herniation. METHODS: Sixty rabbits were divided into blank group (10 rabbits), sham operation group (10 rabbits), model group (40 rabbits) according to method of random digits table. Then model group was divided into made group (group A, 10 rabbits), normal saline group (group B, 10 rabbits), aescin natrium group (group C, 10 rabbits) and danshen and magnesium sulfate injection group (group D,10 rabbits). After model success of lumbar intervertebral disc herniation, different drugs were given to rabbits with lumbar intervertebral disc herniation by ear margin vein. The rabbits of the group C,B,D were respectively given aescin natriu (0.5 mg/kg), normal saline (5 ml/kg), danshen and magnesium sulfate injection (2.0 mg/kg). The rabbits of blank group, sham operation group and group A were not given any disposal. The IgG and IgM level in serum of different groups were determined with ELISA method in fourteen day after model success and drugs given. RESULTS: The level of IgG and IgM in the group A was higher than that of blank group and sham operation group (P < 0.05). The level of IgG and IgM in the group D was lower than that of group B,C (P < 0.05). CONCLUSION: The mixed liquor of danshen () and magnesium sulfate injection could inhibit inflammatory reaction caused by autoimmune response of lumbar intervertebral disc herniation,which provides a new alternative for lumbar intervertebral disc herniation. PMID- 19902763 TI - [Meridian differentiation of lumbar intervertebral disc herniation]. AB - Based on the location of disease and its clinical manifestations, the symptoms and signs of lumbar intervertebral disc herniation are categorized to the pathological changes of Du Meridian, Gallbladder Meridian of Foot Shaoyang, Bladder Meridian of Foot Taiyang, and Kidney Meridian of Foot Shaoyin. As to differentiations, they can be classified into blood stagnation syndrome of Du Meridian, damp-heat syndrome of Gallbladder Meridian, wind-cold-damp obstruction syndrome of Bladder Meridian, kidney-yang deficiency syndrome. Thus the therapeutic effect could be enhanced by methods of manipulation, acupuncture moxibustion and usus internus of Chinese medicinal and the like under the guidance of meridian differentiation. PMID- 19902764 TI - [Powerful stimulus and manipulation to tenderness point in treating pain caused by soft tissue injury outside vertebral canal]. PMID- 19902765 TI - [Anterior release combined with posterior correction for the treatment of severe scoliosis]. AB - OBJECTIVE: To evaluate the clinical results of anterior spinal release combined with posterior correction for the treatment of severe scoliosis. METHODS: Twenty three patients of severe scoliosis were retrospectively analyzed from July 2000 to January 2007. There were 12 males and 11 females with an average age of 15.3 years (ranging from 9 to 18 years). Including 9 congenital scoliosis, 13 idiopethic scoliosis and 1 neurofibromatosis scoliosis. The pre-operative coronal Cobb angles of scoliosis were from 810 to 1260 with the mean of 97.4 degrees. RESULTS: The post-operative coronal Cobb angles was for 100-450 (37.4 degrees on average). All patients were followed up for 6-24 months (means 10 months). Two cases occurrenced addition phenomemon in junctional zone. There were no hook displacement and rod breaking at follow-up. CONCLUSION: Anterior relaxation and posterior correction is a safe and effective treatment. It can achieve well clinical results for the treatment of severe scoliosis. PMID- 19902766 TI - [Analysis of the complication and prevention on treatment of osteoporotic vertebral compressive fractures by percutaneous vertebroplasty (PVP)]. AB - OBJECTIVE: To analyze the reasons of the complications,and the measures taken for the prevention and treatment by percutaneous vertebroplasty (PVP) for osteoporotic thoracolumbars vertebral compressive fractures. METHODS: From May 2004 to June 2008, 204 patients with 286 diseased vertebrae underwent PVP under the guidance of C-arm fluoroscopy, and 56 patients with complications included 18 males, 38 females, ranging in age from 58 to 93 years, with an average of 72 years. The condition of leakage of bone cement and complications was observed. RESULTS: The complications related to the leakage of PMMA found in 60 vertebrae in 49 cases: vertebral canal in 5 cases, intervertebral foramina in 3 cases, soft tissue besides vertebrae in 20 cases,intervertebral discs in 15 cases, venous plexus besides vertebrae in 6 cases. The complications no related to the leakage of PMMA found in 7 cases: the nerve roots injuried in 3 cases; the pain of the patient worsened temporarily in 2 cases; the blood pressure of the patient descent temporarily in 2 cases. CONCLUSION: The main reasons of complications are no-integrated of the compressived vertebrae,unstandard skills of injecting,unsuitable opportunity and quantity of injecting PMMA, unsufficient monitoring of C-arm fluoroscopy, the toxicity of PMMA. The measures of prevention and treatment is strict indication, standard skills of injecting, sufficient monitoring of C-arm fluoroscopy, suitable opportunity and quantity of injecting PMMA, electrocardio-monitoring in operation. PMID- 19902767 TI - [Minimally invasive trochanteric antegrade nail fixation for the treatment of intertrochanteric fractures of femur in elder]. AB - OBJECTIVE: To evaluate the clinical effect of minimally invasive Trochanteric Antegrade Nail (TAN) for the treatment of intertrochanteric fractures of femur in elder. METHODS: From January 2005 to June 2008, 80 patients with intertrochanteric fractures of femur were treated with minimally invasive fixation of Trochanteric Antegrade Nail. There were 16 males and 64 females with an average age of 75 years ranging from 65 to 93 years. According to Evans classification, there were 8 cases of type I, 32 of type II, 30 of type III, 6 of type IV and 4 of type V. After the operation of minimally invasive Trochanteric Antegrade Nail fixation,patients begun to do the continuous passive motion (CPM) and active motion of the injured hip joint and knee joint. RESULTS: All patients were followed from 5 to 18 months with an average of 10 months. All factures were healing in time from 10 to 14 weeks with none screw breakage. Proximal screw exiting out about 0.5 cm occurred in 3 patients without obvert symptom. Three patients appeared lower limb deep venous thrombosis and were cured with medicine. According to the scales of Sanders, pain score was 9.08 +/- 1.19, ambulation score 8.85l +/- 1.09, function score 9.10 +/- 1.27, muscle strength score 9.13 +/ 1.18, daily life score 8.91 +/- l.07, X-ray evaluation score 9.34 +/- 1.09, so as the results were excellent in 56 cases, good in 20, poor in 4. CONCLUSION: With the advantage of mini-invasion, operative convenience and stable fixation, minimally invasive TAN fixation is an effective method for intertrochanteric fractures in elder. PMID- 19902768 TI - [Treatment of fracture of multiple ribs with absorbable rib fixed nail and dacron flap in 12 patients]. AB - OBJECTIVE: To introduce the methods of dacron flap application and its indications in treating fracture of multiple ribs, in order to reduce the incidence of the complications of fracture displacement. METHODS: From September 2006 to March 2008, 12 patients with fracture of multiple ribs were treated with absorbable rib fixed nail and dacron flap. Included 8 males and 4 females,the age was from 22 to 51 years with an average of 38.2 years,the operative time was 2 hours to 3 days after injured. All the patients were closed injury and simultaneously accompanied with significant chest pain and chest tightness. 4 cases with dyspnea, blood in sputum and blood oxygen saturation decreased. The X ray showed 3 cases of unilateral fracture and 9 cases of bilateral rib fractures and all cases accompanied with hemopneumothorax. RESULTS: All patients were followed up from 2 to 26 months with an average of 8 months. All the fractures healed. According to clinical criteria, pain, breathing, ribs alignment etc. to observe the effect, 10 cases got excellent result, 1 case good, 1 case poor. CONCLUSION: It is safe and effective to use absorbable rib fixed nails and dacron flap for treating fracture of multiple ribs and especially for the patients of osteoporosis, comminuted fracture or oblique fracture. PMID- 19902769 TI - [Interventional therapy in the treatment of avascular necrosis of femoral head and short-term efficacy]. AB - OBJECTIVE: To evaluate the clinical effect of avascular necrosis of femoral head (ANFH) with interventional therapy in the near future. METHODS: Twenty-eight patients (19 males, 9 females, the age was from 14 to 70 years old with an average of 38 years) with ANFH were treated by catheterization. The thrombolytic drugs, vasodilator drugs and improving microirculative drugs were respectively injected directly into the arteries supplying the femoral head after ultraselection. Contrasted the changing of the arteries suppplying the femoral head between pretreatment and posttreatment, and observed the bone density of the femoral head at 12-36 months after treatment,and analyzed clinical symptoms (hip pain and joint range of motion) improving. RESULTS: The angiography showed the arteries supplying the femoral head were manifold, the coloration or the femoral head were enhanced after interventional therapy. The X-ray showed the bone density of the femoral head gradually recovered of nearly common by 97.2% (35/36). And hip pain and joint range of motion significantly improved than that of before treatment. CONCLUSION: Interventiomal therapy is a safe and effective method to ANFH, which seems to be promising for wide clinical application. PMID- 19902770 TI - [Treatment of thoracolumbar fractures in minimal invasive with percutaneous transpedical interbody bonegrafting]. PMID- 19902771 TI - [Proximal humeral interlocking antatomic plate for the treatment of proximal humeral fracture and bone cyst]. PMID- 19902772 TI - [Quickly remember the brachial plexus in the orthopeadic clinical teaching]. PMID- 19902774 TI - The 220th year of the Medical Society of Delaware. PMID- 19902773 TI - [Progress of lumbar interspinous process spacer]. AB - Lumbar interspinous process spacer, as a important aspect of dynamic stabilization system,through implant a spacer into interspinous process, which can limit the extension of the pathological segment, reserves the physiological activities basically, increases the volume of the spinal canal and intervertebral foramina, decreases the loading of the posterior of disc, and it doesn't affect the motion and the loading of the adjacent segment significantly. From the available documentary, the biomechanical and clinical results of various lumbar interspinous process spacers were optimistically. The purpose of this paper is to review these investigations in respect of lumbar interspinous process spacers. PMID- 19902775 TI - Further thoughts on Universal Health Care. PMID- 19902776 TI - No more Doritos and lobster tails: a case report of life-threatening sublingual hematoma. AB - This case report highlights a life-threatening complication of mild hemophilia A. We report the onset of airway compromise through a massive sublingual hematoma in a 67-year-old male suffering from the mild form of hemophilia A. This case emphasizes the need for prompt medical attention and recognition of potentially serious complications of the disease in patients suffering with even mild form of this bleeding diathesis. PMID- 19902777 TI - IOM warns about physician-pharma conflicts of interest. PMID- 19902778 TI - Microleakage of contemporary composite systems in dentin cavities. AB - This study evaluated microleakage in vitro of self-etch and multi-step, total etch adhesive systems. Ninety extracted non-carious human molars were randomly assigned to nine groups (n=10) and restored with nine different composite systems: Gluma CPS/Charisma, Syntac Sprint/Tetric Ceram, 3M Single Bond/Silux, Admira Bond/Admira, Optibond Solo Plus/Herculite, Trendy Bond/Saremco, Excite/Tetric Ceram, Syntac Sprint/Compoglass and Promt-L-Pop/Silux. Cavities (2 mm diameter/2 mm deep) were prepared on the facial and lingual surfaces of each tooth previously abraded with a silicon-carbide abrasion discs to produce two level dentin surfaces with whole margins in dentin. Each group of composite systems was treated following the manufacturer's instructions. After they were finished and polished, the specimens were subjected to 100 thermal cycles in baths of 4 degrees +/- 2 degrees C and 58 degrees +/- 2 degrees C with 60 seconds of dwell time for each bath. After the thermocycling, the specimens were sealed with acid-resistant varnish, leaving a 1-mm window around the cervical margin interface, and immersed in 1% methylene blue buffered solution for 4 hours. Then the specimens were sectioned longitudinally in half and evaluated under stereomicroscopy at 30 times magnification, using inserted millimeter measurer. Each specimen was measured for dye penetration, thickness of composite filling and distance from the bottom ofcavity to the pulp chamber. The data were analyzed using Kruskal-Wallis Analysis of Variance (ANOVA) and Pearson t- test. Syntac Sprint/Tetric Ceram, 3M Single Bond/Silux, Admira Bond/Admira, Optibond Solo Plus/Herculite and Gluma CPS/Charisma showed the least microleakage at the occlusal part of the cavity. There was no significant difference of microleakage at gingival part of cavity for all tested composite systems and the best sealing ability showed 3M Single Bond/Silux. Self-etch and multi-step, total-etch adhesive systems showed more or less similar microleakage "in vitro" conditions and marginal sealing ability of all tested composite systems was significantly lower at gingival margins of cavities. PMID- 19902779 TI - [TRAb determination in newly detected patients with hyperthyroidism and their prognostic importance]. AB - INTRODUCTION: In more than 50 years since the first demonstration of the immune basis for thyroiditis was done, the autoimmune disease has been identified as a major cause of disfunction of all endocrine organs. Identification of the antithyroid with the radioreceptor assay is widespread in clinical use for differential diagnosis of other thyroid disorders manifesting with the hyperthyroidism. MATERIAL AND METHODS: Our study included 44 patients with new onset Graves disease which was primarily verified according to the objective status of the patients (signs of hyperthyroidism), biochemical parameters (which included thyroid hormones) and detection of the TRAb (TSAb- thyreotropin stimulating antibodies) at the beginning of diagnosis and in the first, second and after the third month since the introduction of antithyroid therapy (propiltiouracil). RESULTS: On the basis of the TSAb concentration level, we found that the response to the therapy was positive in 65.9% ("responders") of the patients and negative in 22.7% ("non responders") of the patients and without therapeutic effect were 11.4% of the patients. During the therapy (after the first month), in the group of non responders we found increased values of the TSAb (p<0.05) and in the group of responders we found reduction of the TSAb levels (p<0.05). CONCLUSION: Our study demonstrated that the therapy with propiltiouracil, after the first month divide patients with positive and negative response, which could change the therapeutic, options in the future. PMID- 19902780 TI - [Characteristics of shigellosis outbreaks in the AP of Vojvodina]. AB - INTRODUCTION: Shigellosis causes around 165 million infections and around 1 million deaths in the world every year. Two thirds of both infections and deaths are among children younger than 10. Shigellosis mainly spreads by direct or indirect contact, but water- and food-borne outbreaks are not rare. MATERIAL AND METHODS: A descriptive epidemiological method was used to analyse characteristics of shigellosis outbreaks in the AP of Vojvodina in the period 1979-2005. RESULTS: During this period 9.083 shigellosis infections were registered, 51,9% of them in 202 shigellosis outbreaks. The leading way of transmission was contact, but water and food-borne outbreaks were also detected The average number of infections in contact outbreaks of shigellosis was small--6.3 infections. The highest average is in water-borne outbreaks--55.1. Most of the outbreaks were those occuring in families (47.5%) and most of them were registered in august. DISCUSSION: Shigellosis has been mandatory reported in the AP of Vojvodina since 1945 and results of outbreak investigation of this disease were analyzed since 1979. In the first half of this period water was the most common way of agent transmission in shigellosis outbreaks, but in recent 15 years the contact became the leading one. In the last 5 years there was no water-borne shigellosis outbreaks. Almost one third of all outbreaks were imported from Montenegro and Croatia by tourists coming back from those countries. Most of the outbreaks are small outbreaks occuring in families, primarily transmitted by contact. Outbreaks in facilities for care of disabled children and elders are also important. Sh. sonnei and Sh. flexneri are the two serotypes identified, the former in 57.4% and the latter in 40.1% outbreaks. CONCLUSION: Shigellosis is a disease occurring in the AP of Vojvodina primarily in small outbreaks inside families, transmitted by contact, most often in August and more often caused by Sh. sonnei. Health promotion and strict use of general measures of disease control and prevention are important. PMID- 19902781 TI - [Radiotherapeutical chromosomal aberrations in laryngeal cancer patients]. AB - INTRODUCTION: The authors present the results of cytogenetic analysis of 21 patients with laryngeal carcinomas diagnosed and treated in the period 1995-2000 at the Institute of Otorhinolaryngology and Maxillofacial Surgery, Clinical Center of Serbia and Clinical Center of Novi Sad. MATERIAL AND METHODS: The patients were specially monitored and the material was analyzed at the Institute of Human Genetics of the School of Medicine in Belgrade as well as in the Laboratory for Radiological Protection of the Institute of Occupational and Radiological Health "Dr Dragomir Karajovic" in Belgrade. RESULTS: The incidence of chromosomal aberrations and incidence of exchange of material between sister chromatids were observed in the preparation of the metaphasic lymphocyte chromosomes of the peripheral blood obtained in the culture. Structural aberrations were found on the chromosomes in the form of breakups, rings, translocations and dicentrics as early as after a single exposure of patients to tumor radiation dose of 2 Gy in the field sized 5x7. Out of the total number of 35 cultivated blood samples obtained from 13 patients, 21 were successfully cultivated and they were proved to contain chromosomal aberrations. Some of the peripheral blood samples failed to show cell growth in vitro due to the lethal cell damages in vivo. DISCUSSION: We have concluded that the number of structural aberrations cannot be used as a biological measure of the absorbed ionizing radiation dose. The presence of aberrations per se is indicative of the mutagenic effect of the ionizing radiation, which was also confirmed in our series on the original model by cultivation of the peripheral blood lymphocytes in the culture of the cells of the volunteer donors upon in vitro radiation. Using the method of bromdeoxyuridylreductase, the increased incidence of SCE as a mutagenic effect was registered. CONCLUSION: It has been concluded that the increase of absorbed radiation dose in vitro leads to prolonged duration of cell cycle in the same conditions, which proves cytostatic effect of radiation. Further fundamental studies are required for clinical implementation of the findings. PMID- 19902782 TI - [Relevance of polymerase chain reaction technique in immunogenetics and histocompatibility]. AB - INTRODUCTION: During the last decade of the previous century, among many of developing sciences, molecular biology had the most important position. The greatest influence on the development of scientific knowledge had been brought by discovery of PCR technique, that showed numerous advantages in comparison to the existing techniques. PCR PROCEDURE: The capability of PCR technique to amplify specific DNA sequences, in a simple, automatic procedure, accelerated the development of many fields in molecular biology researches such as cloning, identification of new genes and pathogenes, determination and quantification of nucleotide sequences, thus opening fields of experimental investigations and its clinical application. PCR technique is widely used in various investigations from anthropological researches to those in molecular biology, from basic genetic investigations to clinical diagnostics. APPLICATION OF PCR PROCEDURE IN THE FIELD OF IMMUNOGENETICS AND HISTOCOMPATIBILITY: In the field of immunogenetics and histocompatibility, there are several HLA genotyping techniques: PCR-SSOP, detection by sequence specific oligonucleotide probes. PCR-SSP, detection by sequence specific primers, RSCA, detection by Reference-Strand Conformation Analysis, SBT-Sequence Based Typing. The development of these techniques has enabled the testing of the HLA alleles as well as detection of shared nucleotide sequences between alleles on the same or/and different loci and the existence of some locus-specific nucleotide sequences in coding (exon) and noncoding (intron) regions. CONCLUSION: The application of PCR technique in immunogenetics made it possible to define genetic polymorphism in HLA system, which contributed to a significant progress in the field of tissue and organ transplantation, population and genetic studies as well as in studies of disease association and HLA alleles. PMID- 19902783 TI - [Immunomodulatory actions of antibiotics]. AB - INTRODUCTION: Antimicrob drugs and immune system interaction has been studied since the pioneer works of Metchnikoff. After the introduction of antibiotics in clinical practice this area has attracted little attention of investigators, because of the lack of standards. This is the reason that the studying of the influence of antibiotics on immune system is still at its beginning. AIM: To point out the immunomodulatory action of some antibiotics on certain components of immune system. METHODS AND RESULTS: The literature findings show that antibiotics express immunomodulatory action on some components of immune system such as fagocytes (polymorphonucleary, macrophages, monocytes), cytokines, immunoglobulines, and on cellular immunity. The principles of antibiotics action on phagocyte are the inhibition of chemotaxis and oxidants production. Macrolides applied for a short time enhance the phagocytic functions while their long use leads to immunosupression. Some cephalosporines and rifampicin in therapeutic doses inhibit the oxydative metabolism, of macrophages. Tetracyclines, clindamycines, chloramphenicol and tobramycin inhibit the synthesis of superoxyd anione. The action of some antibiotics on cytokine and specific antibodies is also important. Cellular immunity can be affected as well. After administration of certain antibiotics it takes 1-2 weeks to reestablish normal cellular immunity, and for other even more. CONCLUSION: There is still no clear standing on real effects of antibiotics on the immune system. Clinicians should search for more information from this new-old field of investigation in order to give more adequate therapy to patients. PMID- 19902784 TI - [Therapeutic approach in patients undergoing percutaneous coronary interventions]. AB - While the performance of percutaneous coronary interventions remains the domain of interventional cardiologists, the management of these patients before, during, and after the procedure is in the domain of general cardiologists, internists and primary care physicians. Therefore, for optimal patient care it is crucial that all engaged physicians should understand the procedural risks, complications and optimal treatment strategy before, during and after the procedure. Before a percutanous coronary intervention, patients with known allergies to iodinated contrast dye should be pretreated with oral corticosteroids and H1-receptor blockers. Diabetic patients as well as patients with renal failure need special care. Hydration is crucial for patients with renal insufficiency in order to minimise the risk of contrast nephropathy. Metformin therapy should be discontinued before the procedure in patients with renal failure in order to avoid lactic acidosis, and it should be reinstituted after the procedure only when normal serum creatine level is confirmed. Double antiplatelet therapy (aspirin plus clopidogrel) should be initiated at least six hours before the procedure. While aspirin therapy after the procedure is life long, the duration of clopidogrel therapy depends on the type of implanted stent (in patients with bare stents implanted clopidogrel should be taken at least 3 - 4 weeks post procedural, and in patients with drug-eluting stents implanted clopidogrel should be taken at least 6 - 12 months after the procedure due to in-stent restenosis prevention). Patients who experience typical anginal pain in a period of one to eight month after percutaneous coronary revascularization are likely to have restenosis, and they should be reevaluated with stress echocardiography and/or repeated coronary angiography. PMID- 19902785 TI - [Ultrasound detection of endometrial fluid in postmenopausal women]. AB - The aim, of this prospective study was to estimate whether the presence of endometrial fluid detected by transvaginal ultrasound investigation was a marker for the pathological changes of the endometrium in postmenopausal women. MATERIAL AND METHODS: 128 postmenopausal women with uterine bleeding and 29 asymptomatic postmenopausal women underwent transvaginal ultrasound investigation, curettage and histopathological investigation of the curettage specimens. RESULTS: There were significantly more asymptomatic women with endometrial fluid collection was found (41,4%) than those with uterine bleeding (7,8%) (p<0,001). We found 4 cases of carcinoma of the endometrium, 4 with hyperplasia and 1 with polyp of the endometrium in women with uterine bleeding and endometrial fluid collection. In the asymptomatic group of women we found 1 case with polyp and 1 with carcinoma of the endometrium. In the presence of endometrial fluid collection the least thickness of the endometrium measured by transvaginal ultrasound was 12 mm in postmenopausal women with carcinoma of the endometrium, 7 mm in women with hyperplasia and 5 mm with polyp. No pathological changes were found below the above values. CONCLUSION: The presence of endometrial fluid detected by transvaginal ultrasonography is a good marker for pathological changes of the endometrium in postmenopausal women if the endometrial thickness is greater than 4 mm. If the endometrial thickness is 4 mm or less, the presence of endometrial fluid is not an indication for further invasive investigation of endometrial cavity, but we must eliminate possible presence of ednexal or cervical malignant disease in some patients. PMID- 19902786 TI - [Blood glucose self-monitoring among patients with diabetes mellitus type 2 in family medicine practice]. AB - INTRODUCTION: Good knowledge of diabetic patients about their disease is often not related with good glycemic control. The aim of this study was to determine the level of application of acquired knowledge about diabetes in recognizing and resolving hypoglycemic and hyperglycemic conditions in patients who did or did not do blood glucose self-monitoring as well as the impact of self-monitoring on HbA1c during education of patients with diabetes type 2. MATERIAL AND METHODS: There were 91 patients with the type 2 diabetes who completed six months education about their disease in four family medicine practices in Tuzla Canton during the period from March to September 2005. The patients who did or did not do self-monitoring with glucometer were interviewed on knowledge about recognizing and resolving hypoglycemia and hyperglycemia by family physician and HbA1c was assessed at the beginning of the education, 3 months after reading the brochure (passive education) and additional 3 months of group (intensive) education. RESULTS: Out of 91 interviewed patients, there were 29 who did self monitoring by glucometer at the beginning of the education, 30 patients during the passive education and 34 after the intensive education. At the beginning of education, regardless of doing self-monitoring, the patients were less able to recognize and resolve hypoglycemia and even less hyperglycemia. At the end of education, their knowledge was better at both recognizing and resolving hypoglycemia (P=0.01) as well as at recognizing (P=0.01) and resolving hyperglycemia (P=0.001). In the patients who did self-monitoring the average value of the HbA1c did not improve significantly (P=0.44) compared to those who did not practice self-monitoring (P=0.10) during education. CONCLUSION: Only one third of patients with type 2 diabetes had done self monitoring with glucometer and although their knowledge about hypoglycemia and hyperglycemia was improved during education, these patients did not have improved significant values of the HbA1c compared to patients who had not done self-monitoring. PMID- 19902787 TI - [Treatment of pregnancy related venous thromboembolism]. AB - INTRODUCTION: Prevention and treatment of venous thromboembolism during pregnancy are complicated since the use of antithrombotic drugs carries a certain risk to the mother, the fetus or both. Coumarins cross the placental barrier and may be responsible for bleeding, teratogenicity and central nervous system abnormalities. The risk of embriopathy is particularly high between 6 and 12 weeks of gestation. TREATMENT: Heparin is the treatment of choice for thrombosis during pregnancy because it is entirely safe for the fetus, unlike oral anticoagulants. The frequency of heparin-induced thrombocytopenia and osteoporosis is significantly lower if LMWH is applied, so this heparin type is preferable to UFH during pregnancy. Treatment of women with VTE during pregnancy, especially those with thrombophilia, requires individualized dosing and duration of antithrombotic therapy. PERIPARTAL MANAGEMENT: In order to avoid the peripartum anticoagulant heparin effect and possible bleeding, heparin should be discontinued prior to the delivery and reintroduced after the parturition. PROPHYLACTIC REGIMEN: Prophylactic antithrombotic regimen during subsequent pregnancies should also be individualized. The use of low molecular weight heparins is becoming more widespread. They have reliable pharmacokinetics, require less frequent injections than unfractionated heparin and carry a lower risk of treatment complications. LMW heparins are safe and effective and they are replacing UFH as the anticoagulant of choice during pregnancy. Both UFH and LMWH are not secreted into breast milk and can be safely given to nursing mothers. Warfarin does not induce an anticoagulant effect in the breast-fed infant, so it can be safely used in women who require postpartum anticoagulant therapy. PMID- 19902788 TI - [Relations of aqueduct with some structures of mesencephalon]. AB - INTRODUCTION: Aqueductus mesencephali is the biggest part of the ventricular system and that is why it is the most common place of intraventricular obstruction of cerebrospinal fluid. This study was done in order to study topographic characteristics of aqueduct more thoroughly. MATERIALS AND METHODS: Transversal sections of mesencephalon were made in three levels. The first section was made caudally immediately from the posterior commissure. The second section was made in the middle part of the superior colliculi, and the third section was made in the rostral parts of the caudal sections of the superior colliculi. Distances of the aqueduct from structures of mesencephalon, obtained on the second section, are: 1. The distance of the aqueduct from the superior colliculi - 6.96 mm; 2. The distance of the aqueduct from the red nucleus - 6.02 mm; 3. The distance of the aqueduct from the substantia nigra - 12.29 mm; 4. The distance of the aqueduct from the interpeduncular fossa - 10.22 mm. CONCLUSION: Knowledge of the anatomy of the aqueductus mesencephali is very important because of interpretation of patogenesis of hidrocefalus as well as of other syndromes that occure in some pathological processes in the system of ventricles. PMID- 19902789 TI - [Forensic importance of homicide]. AB - INTRODUCTION: This study encompassed the total number of homicides in Bosnia and Herzegovina (B&H) in the period from 1st January 1997 to 31st December 2006 and then analysed homicidal behaviour. The aim is to assess the differences between the people who committed violent and those who committed accidental homicide in Bosnia and Herzegovina. MATERIAL AND METHODS: In a multicentric, retrospective study of comparing the groups with equal number of respondents we analysed the individuals who had committed violent (n=135) and accidental homicides (n=135). The homicides were tested by using sociodemographic and psychosocial items. Measurement instruments were: General data list, Eysenck Personality Questionnaire (EPQ), Hamilton Depression Rating Scale (HDRS), Emotions Profile Index (EPI). The descriptive and multivariable logistic analysis was done statistically. RESULTS: In the descriptive analysis the socio-demographically violent murderers were: male gender (chi 2=3.340, P=0.009), more workers than officials (chi 2=7.340, P=0.011), fathers were more often workers/farmers (chi 2=1.430, P=0.046), gambling (chi 2=13.100, P=0.001) and possible recidivism (chi 2=6.770, a P=0.001). The accidental murderers were family people (chi 2=4.100, P=0.041), with more frequent drug abuse (chi 2=3.190, P=0.012) and they would not repeat the delict. In the multivariate analysis the violent murderers were highly discriminated (P=0.001) from accidental ones by: war involvement r=0.1148, OR=2.971 (95%), CI=1.040-7.890; age, father's education, psychoticism (EPQ) r = 0.1085, OR=0.291 (95%), CI=0.110-0870, HDRS-total r=-0.1797, OR=0.830 (95%), CI=0.710-0.930, destructiveness r=0.1270, OR=1.560 (95%), (CI=1.197-2.032, and deprivation in the P. I. E. tests. CONCLUSION: By the violence of their acts murderers confirm micro-social model of transferring the violence, and transition and heredity confirm the ecological-developmental trans-generation model of violence. Accidental murderers commit homicide in anomy, with intoxications and prolonged psycho-traumatism. PMID- 19902790 TI - Surgical treatment of big splenic artery aneurysm--case report. AB - An aneurysm has been defined as a permanent local dilatation of the diameter of an artery by at least 50% of its normal value. A splenic artery aneurysm is most frequently a visceral artery aneurysm and clinically it is usually asymptomatic but potentially life-threatening at the same time, with the incidence of its rupturing being 2-10% and then the mortality rate ranges from 20 to 36%. A 51 year-old female patient was admitted to the Department of Vascular and Transplantation Surgery in Novi Sad having been found to have a big splenic artery aneurysm during the ultrasound examination of her abdomen after cholecystectomy. The additional diagnostic procedure--computerized tomography of the abdomen with i.v. contrast subtraction angiography--confirmed the splenic artery aneurysm to have the diameter of 5 cm and therefore the elective surgical treatment was indicated after the preoperative preparation and risk assessment. The aneurysm was exposed through Chevron incision, and the detailed surgical exploration was done after the omental bursa had been opened. The aneurysmectomy and the reconstruction of the splenic artery by the termino-terminal anastomosis were performed after the weakening of the wall had been verified. The biopsies of the liver and the aneurysmal sac were done during the surgery. The pathohistological finding confirmed the atherosclerotic etiology of the aneurysm. Since the postoperative course was normal, the patient was discharged on the eighth postoperative day. PMID- 19902791 TI - [Kell immunization--a case report]. AB - INTRODUCTION: The frequency of occurrence of RhD alloimmunization, due to preventive protocols, is decreased in our country, but more often there are other antigens that emerge as a cause of hemolytic disease of fetus. The most prominent is Kell antigen, which promotes specific course of disease based on an innate pathogenetic mechanism. Anti-Kell antibody production is, just as in other atypical antibodies, provoked with transfusion of incompatible blood. Except for the immune-mediated hemolysis, anti-Kell antibodies can also inhibit the function of progenitor (erytroid and megakariocyte) cell lines. CASE REPORT: We present the case of G1P1 woman in whom a distinct fetal hydrops was sonographically detected in the 28th week of pregnancy. The results of immunological tests undoubtedly pointed to Kell immunization (anti-Kell antibody titer was more than 1:32), and antenatal tests for evaluation of fetal condition (Doppler ultrasound and CTG) clearly showed the severe form of hemolytic disease. We concluded that the fetus was in a hopeless, terminal stage of the disease, and then decided to terminate the pregnancy. CONCLUSION: The only clinical approach to a problem of Kell alloimunization is active one. Early cordocentesis is recommended as the optimal method for evaluation of fetal condition. The clinical outcome of the fetus will strictly depend on a timely intrauterine transfusion (IUT) procedure. Prophylaxis emerges as a crucial factor in prevention of Kell-alloimmunization. It is to be considered that all females in childhood and throughout the reproductive period should take only K1-negative blood transfusion in order to decrease the incidence of Kell-alloimmunization. PMID- 19902792 TI - [Patient suitability for cataract surgery under local anesthesia]. AB - In the past two decades local anesthesia has emerged as a reliable, safe and cost effective first choice for cataract surgery in adults. However, the technique is not totally devoid of inherent risks, and requires not only an experienced and well-trained surgeon but also a cooperative patient. Thus, only the patient who is able to communicate, lie still in the supine position, and tolerate their face being covered by a drape is a suitable candidate for local anesthesia. In addition, the preoperative assessment should include particular features of the globe and orbit which may affect the operation as well as a history of allergy to local anesthetics. Finally, it is important to respond to patient preferences and wishes. A careful, patient selection for local anesthesia reduces the risk of surgical cancellation or delay and is essential for patient satisfaction and a successful visual outcome following cataract surgery. PMID- 19902793 TI - [Health workforce in Serbia in two periods: before and after 1950]. AB - INTRODUCTION: Health workers are the embodiment of the core values of the health system, and they guide towards the most effective and economic utilization of all other resources. Historical developmental evidences can be helpful to understand their actual position in Serbia. Description of the health workforce development and coverage in two periods: before and after 1950 in Serbia. MATERIAL AND METHODS: This retrospective study is a descriptive analysis of the health workforce development, based on literature review and routinelly collected data. RESULTS: Health care workforce in Serbia before 1950 was inadequate in number and skills required with regard to the society needs. At the same time health workers were highly motivated to learn and were ready to fit and stretch when needed. Since 1950 onwards there has been a continual intention to enlarge the number and skills of healthcare workforce, but other mechanisms for capacity building of health profession have been often overseen. DISCUSSION: Though health needs were recognized as criteria for health manpower planning and development, they were not appropriately followed in the practice. CONCLUSION: Planning and development of health workforce were influenced by specific political, social and economic conditions in Serbia. PMID- 19902794 TI - Food safety: pathogen transmission routes, hygiene practices and prevention. PMID- 19902795 TI - Survey of food-hygiene practices at home and childhood diarrhoea in Hanoi, Viet Nam. AB - A cross-sectional study was conducted to investigate the potential factors of food-hygiene practices of mothers on the prevalence of diarrhoea among their children. Mothers who had children aged 6 months-5 years were recruited in a hamlet in Viet Nam. The food-hygiene practices included hand-washing, method of washing utensils, separation of utensils for raw and cooked food, and the location where foods were prepared for cooking. A face-to-face interview was conducted, and data on 206 mothers were analyzed. The risk of diarrhoea was significantly higher among children whose mothers prepared food for cooking somewhere other than the table (typically on the ground) compared to children whose mothers prepared food on the table (adjusted odds ratio = 2.85, 95% confidence interval 1.11-7.28). The results indicate that food-hygiene practices of mothers, such as avoiding preparing food for cooking on the ground, has a potential impact in preventing diarrhoea among children in Viet Nam. PMID- 19902796 TI - Phenotypic characterization of Salmonella typhimurium isolates from food-animals and abattoir drains in Buea, Cameroon. AB - Salmonella spp. have been extensively incriminated worldwide as common causes of bacterial gastroenteritis in humans, with food-animals serving as important reservoirs. The study was aimed at investigating cattle and pigs slaughtered in Buea as reservoirs of Salmonella Typhimurium and the susceptibility of isolates to antibiotics. In total, 230 specimens (comprising 50 each from the rectum, ileum, and gall bladder of cattle; and 10 each from same anatomical sites of pigs and 50 from abattoir drains) were analyzed for Salmonella using the standard microbiological, biochemical and serological techniques. Antibiotic susceptibility of the isolates was determined by the Kirby-Bauer disc-diffusion test. The isolates were characterized into biotypes using the API 20E kit, and results were analyzed using the chi-square test. Seventy-five (32.6%) of the 230 specimens were positive for S. Typhimurium, with pigs and abattoir drains presenting the highest level of isolation (40%). Biochemical typing grouped the isolates into five biotypes. Biotype I was the most prevalent (30.6%) while biotype IV was the least prevalent (9.3%) and was absent in samples from pigs. Antibiotic susceptibility studies revealed 14 antibiotypes based on antibiotics used in the study. The predominant antibiotype AMX DOX CEF was recorded in 13 (17.3%) of the isolates. Multidrug resistance (to four or more antibiotics) was recorded in 50.7% (38/75) of the isolates. The most active drugs were ciprofloxacin (98.6%), ofloxacin (93.3%), amikacin (90.6%), and gentamicin (84%). All the isolates (100%) were resistant to tetracycline and ampicillin. Cattle and pigs were found to be reservoirs of S. Typhimurium in the environment of Buea, Cameroon, implying that foods from these sources, if not properly handled, could serve as vehicles for its transmission to humans. PMID- 19902797 TI - Effect of prenatal zinc supplementation on birthweight. AB - Although iron and zinc deficiencies are known to occur together and also appear to be high in Ghana, a few supplementation studies addressed this concurrently in pregnancy. In a double-blind, randomized controlled trial, 600 pregnant women in Ghana were randomly assigned to receive either a combined supplement of 40 mg of zinc as zinc gluconate and 40 mg of iron as ferrous sulphate or 40 mg of elemental iron as ferrous sulphate. Overall, there was no detectable difference in the mean birthweight between the study groups, although the effect of iron zinc supplementation on the mean birthweight was masked by a strong interaction between the type of supplement and the iron status of participants [F (1,179) = 5.614, p = 0.019]. Prenatal iron-zinc supplementation was effective in increasing the mean birthweight among anaemic and iron-deficient women but not among women with elevated iron stores in early pregnancy. PMID- 19902798 TI - Zinc deficiency: descriptive epidemiology and morbidity among preschool children in peri-urban population in Delhi, India. AB - Community-based data relating to factors influencing zinc deficiency among preschool children in India are inadequate. Data of a large, double-blinded, randomized, controlled zinc-supplementation trial were used for assessing the descriptive epidemiology of zinc deficiency among children aged 6-35 months (n = 940). In total, 609 children were followed up for 120 days for information on morbidity. Of these children, 116 from the control group belonging to the upper and the lower 25th quartile of plasma zinc status at baseline were selected for assessing the association of zinc deficiency with prospective morbidity. At baseline, demographic, socioeconomic and dietary information was collected, and anthropometric measurements and levels of plasma zinc were assessed. At baseline, 73.3% of the children were zinc-deficient (plasma zinc < 70 microg/dL), of which 33.8% had levels of plasma zinc below 60 microg/dL. A significantly higher risk of morbidity was prevalent among the subjects with lower plasma zinc compared to those with higher levels of plasma zinc. PMID- 19902800 TI - An outbreak of cholera associated with an unprotected well in Parbatia, Orissa, Eastern India. AB - In November 2003, an outbreak (41 cases; attack rate-4.3%; no deaths) of severe diarrhoea was reported from a village in Orissa, eastern India. Thirteen of these cases were hospitalized. A matched case-control study was conducted to identify the possible exposure variables. Since all wells were heavily chlorinated immediately after the outbreak, water samples were not tested. The cases were managed symptomatically. Descriptive epidemiology suggested clustering of cases around one public well. Vibrio cholerae El Tor O1, serotype Ogawa was isolated from four of six rectal swabs. The water from the public well was associated with the outbreak (matched odds ratio: 12; 95% confidence interval 1.2-44.1). On the basis of these conclusions, access to the well was barred immediately, and it was protected. This investigation highlighted the broader use of field epidemiology methods to implement public-health actions guided by epidemiologic data to control a cholera epidemic. PMID- 19902799 TI - Simplified dosing of gentamicin for treatment of sepsis in Bangladeshi neonates. AB - Extended-interval dosing of gentamicin has several advantages over conventional multiple-daily dosing for the treatment of sepsis. The study was conducted to evaluate the pharmacokinetics of gentamicin for the treatment of neonatal sepsis in predetermined doses at 24- or 48-hour intervals, according to weight category, and to develop a simplified protocol for use in peripheral healthcare settings in developing countries. This prospective observational study was conducted among 59 neonates admitted to the Special Care Nursery at Dhaka Shishu Hospital, Bangladesh, with suspected sepsis and treated with antibiotics, including gentamicin. Intravenous dosing of gentamicin according to weight category was: 10 mg every 48 hours if the infant weighed < 2,000 g (n = 23), 10 mg every 24 hours if the infant weighed 2,000-2,249 g (n = 12), or 13.5 mg every 24 hours if the infant weighed 2,500-3,000 g (n = 24). Peak and trough concentrations of gentamicin and the presence of signs of nephrotoxicity and ototoxicity were determined. The mean +/- standard deviation peak concentration of gentamicin was 12.3 +/- 3.7 microg/mL in infants weighing < 2,000 g, 9.6 +/- 3.1 microg/mL in infants 2,000-2,249 g, and 10.0 +/- 3.4 microg/mL in infants 2,500-3,000 g. Initial peak concentration of gentamicin was > 12 microg/mL in 28.8% and initial trough concentration was > 2 microg/mL in 6.8% of the subjects. No signs of nephrotoxicity or ototoxicity were detected. Favourable pharmacokinetic parameters found with the simplified dosing regimen suggest that it is safe for the treatment of neonatal sepsis. PMID- 19902801 TI - Spatial distribution of diarrhoea and microbial quality of domestic water during an outbreak of diarrhoea in the Tshikuwi community in Venda, South Africa. AB - Total microbial quality assessment and geographical information system were used for evaluating the quality of water and the spatial distribution of diarrhoea cases in Tshikuwi, a rural community in South Africa, during an outbreak of diarrhoea. The water-abstraction points included two groundwater storage tanks, namely Tank 1 and Tank 2 and the Khandanama river. Indicator microbial counts for total coliforms, faecal coliforms, enterococci, and heterotrophic bacteria exceeded the limit for no risk as stipulated by the South African water-quality guidelines for domestic use for Tank 1 and the Khandanama river. Vibrio, Salmonella, and Shigella species were prevalent in the Khandanama river. The spatial distribution of diarrhoea cases showed a hot-spot of diarrhoea cases close to Tank 1 and the Khandanama river. Results of chi-square analysis showed that the proportion of infection from each water source was different or that infection depends on the type of water source (alpha = 0.05). The demonstrated spatial clustering of diarrhoea cases might have been influenced by the poor microbial quality of water used from Tank 1 and the Khandanama river. The results further highlight the urgent need of water-treatment facilities and monitoring of water quality in rural communities of South Africa. PMID- 19902802 TI - Retrospective surveillance for intussusception in children aged less than five years in a South Indian tertiary-care hospital. AB - To facilitate the assessment of the safety profile of rotavirus vaccines effectively, baseline data on intussusception are important for comparison with intussusception rates following the introduction of vaccine. The aim of the study was to describe epidemiological and clinical features of intussusception in children aged less than five years in an Indian medical facility. Hospital data on intussusception for children discharged during 1 January 2001-30 June 2004 from the Christian Medical College Hospital, Vellore, India, were reviewed. Relevant information was extracted from medical records to classify cases according to the criteria of the Brighton Collaboration Intussusception Working Group. Complete review of medical records for clinical and demographic information was only performed for those cases fulfilling level 1 diagnostic certainty (definite intussusception) (Study ID 101245). During the surveillance period, 31 infants and children with definite intussusception were identified. The majority (61.2%) of the cases occurred in the first year of life. The male : female ratio was 3.4 : 1. Intussusception cases occurred round the year with no distinct seasonality. No intussusception-associated death was recorded. This study provides baseline data on intussusception in South India. Cases identified in the study were similar in presentation and demographics as those observed in other Asian settings. Prospective surveillance systems, using standardized case definitions will further increase the understanding of the aetiology and epidemiology of intussusception, especially as new rotavirus vaccines are made available. PMID- 19902803 TI - Demographic and health-related risk factors of subclinical vitamin A deficiency in Ethiopia. AB - The study was conducted to determine the demographic and health-related risk factors of subclinical vitamin A deficiency in Ethiopia. Blood samples were collected from 996 children in 210 clusters across the nation for analysis of serum retinol. Interviews were conducted with the respective mothers of the 996 children on presumed risk factors of vitamin A deficiency. A higher subclinical vitamin A deficiency was associated with: not receiving vitamin A supplement over the year, having been ill during the two weeks preceding the survey, no or incomplete vaccination, belonging to a mother with high parity, and low levels of awareness of vitamin A. Moreover, being from Muslim household was strongly associated with higher levels of subclinical vitamin A deficiency. Among the risk factors identified, low levels of vaccination, high parity, and low levels of maternal awareness of vitamin A contributed to higher risks of vitamin A deficiency among Muslim children. The findings underscore the need for creation of strengthened awareness of family planning and importance of vitamin A, promotion of vaccination and child health, intensification of vitamin A supplementation, and in-depth investigation on factors contributing to increased vulnerability of Muslim children. PMID- 19902804 TI - Technical and social evaluation of arsenic mitigation in rural Bangladesh. AB - Technical and social performances of an arsenic-removal technology--the sono arsenic filter--in rural areas of Bangladesh were investigated. Results of arsenic field-test showed that filtered water met the Bangladesh standard (< 50 microg/L) after two years of continuous use. A questionnaire was administrated among 198 sono arsenic filter-user and 230 non-user families. Seventy-two percent of filters (n = 198) were working at the time of the survey. Another 28% of the filters were abandoned due to breakage. The abandonment percentage (28%) was lower than other mitigation options currently implemented in Bangladesh. Households were reluctant to repair the broken filters on their own. High cost, problems with maintenance of filters, weak sludge-disposal guidance, and slow flow rate were the other demerits of the filter. These results indicate that the implementation approaches of the sono arsenic filter suffered from lack of ownership and long-term sustainability. Continuous use of arsenic-contaminated tubewells by the non-user households demonstrated the lack of alternative water supply in the survey area. Willingness of households to pay (about 30%) and preference of household filter (50%) suggest the need to develop a low-cost household arsenic filter. Development of community-based organization would be also necessary to implement a long-term, sustainable plan for household-based technology. PMID- 19902805 TI - Evaluating the experience of GAPS--a methodology for improving quality of mass immunization campaigns in developing countries. AB - The existence of pockets of under-vaccinated persons has allowed outbreaks of disease in countries that have achieved high levels of vaccination coverage. A field-based methodology--GAPS (Geographic Assessment of Planning and Services)- was developed to predict, in advance of an immunization campaign, the sites of which are most likely to have a pocket of unvaccinated persons and then use this information to improve planning, supervision, and evaluation of the campaign. At this time, there have been two applications of GAPS (Nepal and Ethiopia). The purpose of this paper was to evaluate these two applications of GAPS and make recommendations regarding its future use. Structured, expert interviews were conducted with at least three campaign organizers to evaluate each application of GAPS using purposive sampling. An evaluation of an individual campaign was considered positive when at least two of the three campaign organizers considered GAPS to be useful and worthwhile. The three campaign organizers interviewed following the GAPS application in Ethiopia responded that GAPS was useful and worth the effort. In Nepal, all four campaign organizers responded that GAPS was useful and worth the effort. Some suggestions for improvement were also identified. Although this evaluation was limited in the number of applications evaluated, GAPS appears to have promise as a practical method to help improve the quality of mass immunization campaigns. And even if no pockets of unvaccinated persons are found, the method may serve as a rapid quality-check of administrative estimates of coverage. Further applications in different settings are needed to confirm these findings or under what circumstances GAPS might best be used. GAPS may also be considered for improving other types of health campaigns, such as distribution of insecticide-treated bednets, vitamin A capsules, and deworming medications. PMID- 19902806 TI - Household decision-making about delivery in health facilities: evidence from Tanzania. AB - This study investigated how partners' perceptions of the healthcare system influence decisions about delivery-location in low-resource settings. A multistage population-representative sample was used in Kasulu district, Tanzania, to identify women who had given birth in the last five years and their partners. Of 826 couples in analysis, 506 (61.3%) of the women delivered in the home. In multivariate analysis, factors associated with delivery in a health facility were agreement of partners on the importance of delivering in a health facility and agreement that skills of doctors are better than those of traditional birth attendants. When partners disagreed, the opinion of the woman was more influential in determining delivery-location. Agreement of partners regarding perceptions about the healthcare system appeared to be an important driver of decisions about delivery-location. These findings suggest that both partners should be included in the decision-making process regarding delivery to raise rates of delivery at facility. PMID- 19902807 TI - Seroprevalence of syphilis among a cohort of HIV-infected subjects in North Central Nigeria. PMID- 19902808 TI - Bell's palsy as a possible complication of hepatitis B vaccination in a child. AB - Bell's Palsy is the sudden onset of unilateral temporary paralysis of facial muscles resulting from seventh cranial nerve dysfunction. Presented here is a two year old female patient with right peripheral facial palsy following hepatitis B vaccination. Readers' attention is drawn to an uncommon cause of Bell's Palsy, as a rare complication of hepatitis B vaccination. PMID- 19902809 TI - The need for broad spectrum and safe anti-epileptic drugs in childhood epilepsy. AB - In this short review, the challenges in the treatment of children with epilepsy are highlighted. It is argued that in many cases, broad spectrum anti-epileptic drugs are the best and logical choice at the start of the treatment. At this moment, valproate, lamotrigine, topiramate, levetiracetam and possibly zonisamide can be considered as safe broad spectrum anti-epileptic drugs. PMID- 19902810 TI - Brain renin-angiotensin system in cognitive function: pre-clinical findings and implications for prevention and treatment of dementia. AB - Biochemical, physiological and functional studies suggest that the brain renin angiotensin system (RAS) is regulated independently of the peripheral RAS. Also, all enzymes and peptides necessary for the biosynthesis of various angiotensins have been recognized within the central nervous system. There are divergent opinions concerning the localization of the different components of this system which is not fully understood. It is believed that central actions of angiotensins are not exclusively associated with their traditional roles, many studies showing that central angiotensins are also involved in learning and memory processes. Moreover, clinical trials and studies on animal models suggest that pharmacological manipulation of angiotensin ligands may be of clinical importance in slowing or even stopping the cognitive deterioration seen in vascular dementia and Alzheimer's disease. PMID- 19902811 TI - Ischemic stroke and hyperhomocysteinemia: truth or myth? AB - Hyperhomocysteinemia is generally acknowledged as a treatable risk factor for atherotrombotic diseases, but a causal relationship between both is not yet definitively established. Hyperhomocysteinemia originates from a deviation in the methionine-homocysteine metabolism including disturbances of enzymes, vitamin deficiencies and different other factors. Observational studies, genetic polymorphism studies and several meta-analyses implicate already a causal relation between homocysteine and cerebrovascular diseases. It is useful to determine homocysteine levels for stroke who present no clue for vascular disease and thrombosis, who have an ischemic stroke at a young age and who have a family history of premature atherosclerosis. Because of the low cost and safety of the therapy, the American Heart and Stroke Association advises to treat patients with a stroke and hyperhomocysteinemia daily with 0,4 mg folic acid, 2,4 microg vitamin B12 and 1,7 mg vitamin B6. A significant benefit in secondary prevention is not yet proven. The results of larger follow-up trials have to be published. PMID- 19902812 TI - Treatments for progressing Parkinson's disease: a clinical case scenario study. AB - OBJECTIVE: A 'case scenario' study on clinical decisions in progressing Parkinson's disease (PD) was developed to complement scientific evidence with the collective judgment of a panel of experts. METHODS: The opinions of 9 experts in movement disorders on the appropriateness of 9 common pharmacological treatments for 33 hypothetical patient profiles were compared to those of 14 general neurologists. Before rating the case scenarios, all participants received a document integrating European and US guidelines for the treatment of patients with advanced PD. Case scenarios showing disagreement or with inconsistencies in appropriateness ratings were discussed at a feedback meeting. A tool for interactive discussion on the clinical case scenarios included was developed based on the outcome of the study. RESULTS: Current guidelines are often insufficient to adequately guide the management of patients with progressing PD. The case scenario study did not reveal major differences in opinions between experts in movement disorders and general neurologists about the appropriateness of certain drug choices for specific case scenarios. However in about 1 out of 5 treatment decisions where experts stated appropriateness or inappropriateness, the general neurologists panel had no or dispersed opinions. CONCLUSIONS: This study reveals more uncertainty about treatment of advanced PD in general neurologists compared with experts in movement disorders and underlines the need for additional support for guiding treatment decisions in clinical practice. PMID- 19902813 TI - Repetitive transcranial magnetic stimulation in patients with progressive supranuclear palsy: a pilot study. AB - Progressive supranuclear palsy is a progressive neurodegenerative disorder for which no specific treatment is known at present. In this report we treated a small group of clinically diagnosed patients with rapid-rate repetitive transcranial magnetic stimulation of the motor cortex for five days. This resulted in modest and transient improvements, especially of the axial symptomatology. Side-effects were not reported. PMID- 19902814 TI - Lack of association between the paraoxonase 1 Q/R192 single nucleotide polymorphism and stroke in a Chinese cohort. AB - BACKGROUND: Serum paraoxonase (PON1) is an HDL-associated esterase that hydrolyzes products of lipid peroxidation and prevents the oxidation of LDL. Paraoxonase 1 (PON1) was implicated in susceptibility to stroke in previous studies. We investigated the correlation between the paraoxonase Gln-Arg 192 polymorphism (PON1Q/R192) and stroke including cerebral hemorrhage and cerebral infarction. METHOD: The association between the paraoxonase Gln-Arg 192 polymorphism (PON1Q/R192) and stroke was investigated in 1019 subjects, which involved 305 cases with cerebral hemorrhage, 375 cases with cerebral infarction and 339 healthy controls. RESULTS: The PON1Q/R192 genotype distribution in the cerebral hemorrhage group was QQ13.1%, QR48.2% and RR38.7% and in the cerebral infarction group was QQ13.6%, QR44.0% and RR42.4% respectively. There was no significant difference in PON1Q/R192 allele and genotype distribution between the patient group and the control group (P > 0.05). The PON1 polymorphism was not associated with cerebral hemorrhage or infarction. CONCLUSION: Our study suggests that serum paraoxonase (PON1) is not associated with cerebral hemorrhage or infarction, although it is a lipolactonase which is associated with HDL apolipoprotein A-I (HDL-apoA-I) and plays a role in the prevention of atherosclerosis. PMID- 19902815 TI - Sciatic injection injuries in adults: is dipyrone a foe to nerve? AB - Sciatic nerve injury is an iatrogenic and rare complication of intragluteal injections. There are a few reports on the subject in adults. Data were collected for eight years from patients referred to our electroneuromyography laboratory. Twenty-eight adult patients (20 males and 8 females) diagnosed with post injection injuries were identified by history, clinical and electrophysiological findings. A complete history was available in 26, all reporting sudden pain and subsequent radiation of pain and numbness in the distribution of the sciatic nerve. In 17 of the 28 the common peroneal portion was affected more severely than the posterior tibial portion; in seven the opposite. Twenty-three patients were able to name the injected drug, and dipyrone (metamizole) specifically, as the responsible agent in 11 of them (47,8%). Injection neuropathy is not specific to children only alone and according to our data special attention is needed during intragluteal injections for thin men and/or usage of dipyrone. PMID- 19902816 TI - Harlequin syndrome: two new cases and a management proposal. AB - The Harlequin syndrome is a rare autonomic disorder, characterized by unilateral diminished sweating and flushing of the face in response to heat or exercise. We present two new cases and evaluate the data of 83 patients described in the literature. We provide diagnostic and therapeutic guidelines. PMID- 19902817 TI - A pattern of otoferlin expression interrupted by gentamicin exposure in ribbon synapse of inner hair cell in C57BL/6J mice. AB - Despite ototoxic effect of aminoglycosides on auditory sensory system has been well documented, the mechanism of ototoxic damage on synaptic connection (ribbon synapse) between inner hair cells (IHCs) and spiral ganglion neurons (SGNs) is still largely unknown. Otoferlin is essential for a late step in exocytosis of neurotransmitter at ribbon synapses. In this study, C57BL/6J mice were intraperitoneally injected with gentamicin (100 mg/kg) once a day, the apical coils of organ of Corti were obtained at the 4th day, the 7th day and the 10th day respectively (n = 10/per group), the normal mice without a Gentamicin exposure were set for control (n = 10/per group). Otoferlin was detected by both immunofluorescence and western blotting. The hearing thresholds were measured by auditory brainstem responses (ABR). The expression of Otoferlin was up-regulated during the period of first 7days with a Gentamicin exposure. The down-regulation of Otoferlin expression was found after the 7th day with a gentamicin exposure. The hearing thresholds showed a linear increase. Gentamicin exposure may enhance transiently the expression of Otoferlin in ribbon synapse in C57BL/6J mice. This property might correlate with the ribbon synaptic plasticity in C57BL/6J mice. It may provide us a new insight for a further detailed study of the ribbon synaptic plasticity under the condition of aminoglycosides application. PMID- 19902818 TI - Susac syndrome: a case report and PET imaging findings. AB - We describe the case of a twenty-year-old woman with subacute encephalopathy, who subsequently developed hearing loss and ophtalmopathy. The clinical triad and typical findings on magnetic resonance imaging and cerebrospinal fluid analysis led to the diagnosis of Susac syndrome. Brain positron emission tomography showed abnormalities which are comparable with other types of central nervous system vasculitis, and distinct from those found in multiple sclerosis. PMID- 19902819 TI - Delayed and prolonged coma after acute disulfiram overdose. AB - We report the case of a 35-year-old man presenting with a delayed and prolonged coma due to an intentional overdose with disulfiram without simultaneous alcohol ingestion. The clinical features--comprising a severe toxic encephalopathy with coma and convulsions, in combination with a quadriparesis outlasting the loss of consciousness--are summarized, and the physiopathology is reviewed. PMID- 19902820 TI - Migraine presenting as chronic facial pain. AB - We report the case of a 44-year-old woman with chronic facial pain. She was treated with several analgesics, prophylactic medications and infiltrations, but all treatment modalities were ineffective. Finally, the diagnosis of medication overuse headache complicating migraine without aura was made and an appropriate treatment was initiated. Migraine is a very common primary headache and rarely presents as isolated facial pain. Stimulation of the dura with activation of the trigeminovascular system can result in pain in any of the three divisions of the trigeminal nerve. This is the anatomic basis of migraine pain presenting as referred pain to the second division of the trigeminal nerve. The atypical presentation of migraine pain can easily lead to inappropriate treatment regimens. PMID- 19902821 TI - Six year survival after prolonged temozolomide treatment in a 30-year-old patient with glioblastoma. AB - Glioblastoma (GBM) is the most malignant primary brain tumour in adults. Since 2005 surgery followed by radiotherapy with concomitant Temozolomide (TMZ) is the standard care for patients with a GBM. Despite these improved treatment strategies, survival of GBM-patients remains poor; and there are very few patients who survive for a long time. Also there is no standard therapeutic strategy after six cycles of TMZ, and further treatment is at the physician's discretion. We report a case of a young patient with a glioblastoma who, not only showed dramatic clinical and radiological improvement after TMZ treatment but who now also (under continued TMZ therapy) survives over 6 years, with complete remission clinically and radiologically. Up till now there are no studies describing TMZ treatment in GBM patients for as long as 6 years. PMID- 19902822 TI - Proximal basilar artery fenestration with bridging artery appearance. PMID- 19902823 TI - Giant arachnoidal cyst. PMID- 19902824 TI - "This must be true--I read about it on the Internet". PMID- 19902825 TI - New drug information: Besivance. PMID- 19902826 TI - Underarm rash worsens with OTC treatment. PMID- 19902827 TI - A day in the life. PMID- 19902828 TI - Beyond the red reflex: examining the eye with an ophthalmoscope. PMID- 19902829 TI - Endovascular embolization with a vascular plug corrects a PAVM. PMID- 19902830 TI - Referral to a dietitian uncovers why this woman cannot lose weight. PMID- 19902831 TI - Poor medication history plus slow symptom onset delays a diagnosis. PMID- 19902832 TI - Understanding the link between HPV and oropharyngeal cancers. PMID- 19902833 TI - How I learned to rate pain. PMID- 19902834 TI - Benefits and harms of widespread PSA test use still unclear. PMID- 19902835 TI - A 17-year-old male with leg weakness and low back pain. PMID- 19902836 TI - A jaundiced view: more than meets the clinician's eye. PMID- 19902837 TI - Case of the month. Compartment syndrome. PMID- 19902838 TI - Streptococcosis in farmed Litopenaeus vannamei: a new emerging bacterial disease of penaeid shrimp. AB - Presumptive systemic streptococcal infections were detected histologically in farmed Litopenaeus vannamei juveniles submitted from a Latin American country and the bacteria isolated. Characterization work demonstrated that the Gram-positive cocci form chains, grow aerobically and anaerobically, are oxidase- and catalase negative, non-hemolytic, non-motile, Lancefield Group B positive and PCR positive when amplified with a universal streptococcal primer set. Differing Streptococcus identifications were obtained using API 20 Strep and Biolog systems, the former identifying the isolate as S. uberis and the latter as S. parauberis. Injection of specific pathogen-free (SPF) L. vannamei with the bacteria resulted in 100% mortality by 3 d post-injection with successful recovery of the agent from moribund test shrimp hemolymph samples. The recovered isolate was used in per os and waterborne exposure studies of SPF L. vannamei with mortalities ranging from 40 to 100% and 80 to 100%, respectively. Histologic analysis of 5 to 8 moribund shrimp from each exposure method demonstrated that all contained a severe bacteremia characterized by numerous free cocci within the hemolymph and aggregates of vacuolated hemocytes with notable intravacuolar cocci. This unique lesion type was most pronounced within the lymphoid organ and considered pathodiagnostic for this disease. Experimentally induced lesions were identical to those in naturally infected farmed shrimp and the Streptococcus sp. responsible was re-isolated, fulfilling Koch's postulates. Five freeze/thaw cycles of 10 experimentally infected shrimp were performed over a 2 mo period and the bacteria successfully cultured from all shrimp at each interval. These collective findings describe the first reported case of streptococcosis in marine penaeid shrimp in the Western Hemisphere and indicate that the agent may be disseminated via live or frozen infected shrimp. PMID- 19902839 TI - Case reports of melamine-induced pathology in penaeid shrimp fed adulterated feeds. AB - Shrimp (Penaeus monodon and P. vannamei) specimens were submitted to the University of Arizona's Aquaculture Pathology Laboratory (UAZAPL) and to the Texas Veterinary Medical Diagnostic Laboratory (TVMDL) in 2007 and 2008 from shrimp-rearing facilities in India and Indonesia for histological testing. These were found to present prominent golden to greenish-brown needle- and plate-like birefringent crystals within multifocal hemocytic granulomas in the antennal gland tubules and peritubular hemal sinuses. Their appearance was very similar to melamine-cyanuric acid-induced crystals previously described from cat and dog kidneys with melamine-associated renal failure (MARF). Significant chronic mortalities were reported from the affected P. vannamei farms in Indonesia, but were not observed in the affected P. monodon facility in India. Shrimp feed was suspected as the source of melamine due to the similarity of the shrimp antennal gland lesions to those present in MARF. 'Normal' and 'suspect' feed samples from the facilities in Indonesia and India were sent to regional laboratories for analysis. Melamine was detected in 2 of 4 feed samples from an affected Indonesian farm. Melamine was not detected in 'normal' feed from the Indian facility, but it was found in 2 'suspect' samples (Feeds A and B) at levels of 183.39 and 112.50 ppm, respectively. A bioassay of Feed A with P. vannamei at UAZAPL confirmed that the melamine-contaminated feed induced prominent granulomas in the antennal gland with the characteristic crystals within 10 d of the first feeding, experimentally confirming the direct relationship of melamine adulterated feed to the unique pathology observed. PMID- 19902840 TI - Shrimp pathogenicity, hemolysis, and the presence of hemolysin and TTSS genes in Vibrio harveyi isolated from Thailand. AB - The virulence factors of Vibrio harveyi, the causative agent of luminous vibriosis, are not completely understood. We investigated the correlations between shrimp mortality, hemolysis, the presence of a hemolysin gene (vhh), and a gene involved in the type III secretion system (the Vibrio calcium response gene vcrD). V harveyi HY01 was isolated from a shrimp that died from vibriosis, and 36 other V. harveyi isolates were obtained from fish and shellfish in Hat Yai city, Thailand. An ocean isolate of V. harveyi BAA-1116 was also included. Thirteen isolates including V harveyi HYO1 caused shrimp death 12 h after injection. Most V harveyi isolates in this group (designated as Group A) caused hemolysis on prawn blood agar. None of the shrimp died after injection with V harveyi BAA-1116. Molecular analysis of all V harveyi isolates revealed the presence of vcrD in both pathogenic and non-pathogenic strains. Although vhh was detected in all V harveyi isolates, some isolates did not cause hemolysis, indicating that vhh gene expression might be regulated. Analysis of the V harveyi HYO1 genome revealed a V cholerae like-hemolysin gene, hlyA (designated as hhl). Specific primers designed for hhl detected this gene in 3 additional V harveyi isolates but the presence of this gene was not correlated with pathogenicity. Random amplified polymorphic DNA (RAPD) analysis revealed a high degree of genetic diversity in all V harveyi isolates, and there were no correlations among the hhl-positive isolates or the pathogenic strains. PMID- 19902841 TI - Antibody responses of turbot Psetta maxima against various antigen formulations of scuticociliates Ciliophora. AB - The kinetics of the antibody production and the protection at challenge were studied in turbot inoculated with various scuticociliate antigen preparations: live ciliates putatively attenuated through long-term in vitro culture (Trial 1) and formalin-killed ciliates without or with GERBU adjuvant in Trials 2, 3, and 4. Antigen used in killed preparations was a mixture of 3 different ciliate isolates (V3) in the case of Trials 2 and 3, whereas in Trial 4, monovalent (V1), trivalent (V3) or pentavalent (V5) antigens were used. A booster injection was administered 28 to 29 d post-priming in all trials. Fish were challenged with virulent live ciliates after the immunization protocol, testing 2 challenge times in Trial 2 (t1 and t2). No protection was obtained in Trial 1 with live ciliates, which in turn were not completely attenuated. Using killed-ciliate formulations, protection was high only in Trial 3 when a low dose (50 000 ciliates fish(-1)) was used for challenge. In Trial 1, heat-inactivated sera of antigen-inoculated fish agglutinated the homologous ciliate, although no specific antibodies were detectable by ELISA. In contrast, high specific antibody levels were detected in antigen-inoculated fish in Trials 2 and 4, and their amount increased progressively, usually peaking after challenge. No advantage was obtained from the use of V5 antigens compared to V1 or V3. No good correlation was observed in most cases between serum antibody levels and protection. Although the use of GERBU adjuvant generally increased the specific immune response, some undesired side effects indicate a need to adjust dosage and/or improve the formulation. PMID- 19902842 TI - Phylogenetic relationship of Perkinsus olseni from the Ebro Delta, Spain, to other Perkinsus species, based on ribosomal DNA sequences. AB - The phylogenetic relationship of Perkinsus olseni originating from the Ebro Delta, Spain, to other Perkinsus spp. was investigated using the nontranscribed spacer (NTS) region and the internal transcribed spacer (ITS) region (including ITS1, 5.8S and ITS2) of the ribosomal DNA sequences. These 2 molecular markers (NTS and ITS) were sequenced from prezoosporangia of Perkinsus sp. originating from Manila clam Ruditapes philippinarum from the Ebro Delta. The sequence of the 5.8S ITS region of the ribosomal RNA gene was 100% similar to that of P. olseni. Higher genetic variability was found for the NTS sequence, with 80.7 to 81.8% similarity to P. olseni. The NTS sequence of a P. olseni isolate previously detected in R. decussatus from the same area was also obtained and showed 81% identity with our isolate. Evidence obtained from phylogenetic analysis of the 5.8S ITS and NTS aligned sequences appears to indicate that P. olseni strains group together according to their host rather than their geographic origins within a well-resolved P. olseni clade. PMID- 19902843 TI - Emerging infectious diseases in cetaceans worldwide and the possible role of environmental stressors. AB - We reviewed prominent emerging infectious diseases of cetaceans, examined their potential to impact populations, re-assessed zoonotic risk and evaluated the role of environmental stressors. Cetacean morbilliviruses and papillomaviruses as well as Brucella spp. and Toxoplasma gondii are thought to interfere with population abundance by inducing high mortalities, lowering reproductive success or by synergistically increasing the virulence of other diseases. Severe cases of lobomycosis and lobomycosis-like disease (LLD) may contribute to the death of some dolphins. The zoonotic hazard of marine mammal brucellosis and toxoplasmosis may have been underestimated, attributable to frequent misdiagnoses and underreporting, particularly in developing countries and remote areas where carcass handling without protective gear and human consumption of fresh cetacean products are commonplace. Environmental factors seem to play a role in the emergence and pathogenicity of morbillivirus epidemics, lobomycosis/LLD, toxoplasmosis, poxvirus-associated tattoo skin disease and, in harbour porpoises, infectious diseases of multifactorial aetiology. Inshore and estuarine cetaceans incur higher risks than pelagic cetaceans due to habitats often severely altered by anthropogenic factors such as chemical and biological contamination, direct and indirect fisheries interactions, traumatic injuries from vessel collisions and climate change. PMID- 19902844 TI - Detection of Panulirus argus Virus 1 (PaV1) in exported frozen tails of subadult adult Caribbean spiny lobsters Panulirus argus. AB - The Caribbean spiny lobster Panulirus argus is a valuable fishing resource and the trade in frozen lobster tails is an important industry. However, the presence of the pathogenic virus Panulirus argus Virus 1 (PaV1), which causes systemic infection in P. argus and is particularly lethal to juvenile individuals, has not been previously examined in imported/exported lobster products. We used PCR assays to determine the presence of PaV1 in abdominal muscle tissue of 22 frozen P. argus tails exported from Belize to Mexico. Based on their size, the tails belonged to subadult-adult lobsters. Using specific primers targeted for PaV1 resulted in 11 tails showing a specific 499 bp band. The sequence of positive amplified fragments showed a high similarity to PaV1 (95% identity with GenBank accession no. EF206313.1). Although the pathogenicity of PaV1 was not evaluated in the present study, our results provide the first evidence of PaV1 in frozen lobster tails exported in the seafood industry as well as the first molecular evidence of PaV1 in adult lobsters. PMID- 19902845 TI - DNA identification of ciliates associated with disease outbreaks in a New Zealand marine fish hatchery. AB - Ciliates associated with fish mortalities in a New Zealand hatchery were identified by DNA sequencing of the small subunit ribosomal RNA gene (SSU rRNA). Tissue samples were taken from lesions and gill tissues on freshly dead juvenile groper, brain tissue from adult kingfish, and from ciliate cultures and rotifers derived from fish mortality events between January 2007 and March 2009. Different mortality events were characterized by either of 2 ciliate species, Uronema marinum and Miamiensis avidus. A third ciliate, Mesanophrys carcini, was identified in rotifers used as food for fish larvae. Sequencing part of the SSU rRNA provided a rapid tool for the identification and monitoring of scuticociliates in the hatchery and allowed the first identification of these species in farmed fish in New Zealand. PMID- 19902846 TI - Outbreak of Sciaenacotyle panceri (Monogenea) on cage-reared meagre Argyrosomus regius (Osteichthyes) from the western Mediterranean Sea. AB - Two different batches of meagre Argyrosomus regius were stocked in the same floating cage located in north-eastern Sardinia (western Mediterranean Sea). After 4 mo, the fish started showing non-specific disease signs, such as lethargy, emaciation, gill anaemia and mortality. In total 65 specimens of meagre of both stocks were sampled for parasitological and microbiological analysis, and only 1 species of parasite, the microcotylid monogenean Sciaenacotyle panceri, was found on the gills of caged meagre. No protozoan or other metazoan parasites were found and no bacteria or viruses were isolated. After the outbreak, a total mortality rate of 5 to 10% was estimated for the older fish stock, while no noteworthy mortality was recorded in younger fish (<2%). This is the first account of S. panceri on the gills of cultured meagre as causative agent of disease and mortality. The effects of the presence of this parasite emphasise the need for disease control strategies of cultured meagre, and investigating the transfer of microcotylids from wild to caged fish. PMID- 19902847 TI - Effect of hyposalinity on the infection and pathogenicity of Miamiensis avidus causing scuticociliatosis in olive flounder Paralichthys olivaceus. AB - Miamiensis avidus, a causative agent of scuticociliatosis in olive flounder Paralichthys olivaceus, was previously reported to proliferate the fastest in media with an osmolarity of 300 to 500 mOsm kg(-1). This suggests that hyposaline conditions can promote the development of the disease. In the present study, olive flounder constantly showed high mortalities when they were experimentally challenged with the parasite by immersion and subsequently reared in hyposaline conditions. Furthermore, affected flounder produced by the challenge showed symptoms identical to those in naturally infected flounder. It was experimentally demonstrated that hyposaline conditions can be a key factor for the development and outbreak of scuticociliatosis in olive flounder. PMID- 19902848 TI - Treatment results in adult-onset recurrent respiratory papillomatosis. AB - PROBLEMS/OBJECTIVES: To assess remission rate, treatment results, and factors for remission in patients with adult-onset recurrent respiratory papillomatosis (AORRP). METHODOLOGY: In this retrospective study, the clinical and pathological data of 51 patients with AORRP, treated in University Hospitals between 1972 and 2006 were reviewed. The male-female ratio was 7:3. At diagnosis, the median age was 43 years, and the median retrospective Coltrera-Derkay Staging and Severity score was 6 (range, 2-28). Twenty-one patients (41%) received only surgical treatment. Thirty patients (59%) were treated with surgery and adjuvant intralesional cidofovir. RESULTS: At the time of analysis in December 2006, 69% of the patients were in remission. Of those, 46% had been treated with adjuvant cidofovir. Of the patients who were not in remission, 87% had been treated with adjuvant cidofovir. This difference was statistically significant (p = 0.005). No significant difference was found between the remission group and the non remission group for the factors age, gender, smoking habits, alcohol habits, GERD, severity and duration of therapy. CONCLUSIONS: AORRP was curable in most patients after long intensive treatment, with a general remission rate of 69%. Cidofovir was a negative factor for remission (p = 0.005). No other statistically significant factors for remission were found. Although a control group was present, it was not possible to perform a randomized study with comparable groups. A well-designed placebo-controlled, double-blinded, randomized trial to assess the outcomes of adjuvant intralesional cidofovir therapy for RRP would be valuable. PMID- 19902849 TI - Surgical findings and post-operative parathormone levels in patients with secondary hyperparathyroidism. AB - OBJECTIVE: The peri-operative and immediate post-operative outcome of secondary hyperparathyroidism treated with subtotal parathyroidectomy is reported. METHODS: We studied 100 patients with chronic renal failure who underwent subtotal parathyroidectomy at our department. Surgical eligibility was based on hyperparathyroidism stage, defined by symptoms of osteodystrophy and/or the presence of hypercalcemia and hyperphosphatemia refractory to medical treatment. Parathormone levels were measured pre-operatively and during the first post operative days. RESULTS: During surgery, four parathyroid glands were identified in 86% of patients, five glands in 1%, and less than four glands in 13%. The ratio of hyperplastic to normal glands was 93:7. No correlation was found between anatomic location of the glands and the presence of hyperplasia. Parathormone decreased to normal or very low values in 93% of the patients. In seven cases, the lowest post-operative parathormone value was above 30 pg/ml, although four glands were removed in four of these patients. In 95% of the patients with four or more identified glands, post-operative serum parathormone levels decreased to normal or very low values. In 23% of the patients with less than four glands, parathormone levels remained too high. On the other hand, post-operative parathormone values normalized in 10 patients who had less than four glands identified during surgery; in two of them, parathyroid tissue was found during postoperative pathological examinations of the resected thyroid lobe. CONCLUSIONS: Subtotal parathyroidectomy is an acceptable treatment in patients with refractory hyperparathyroidism. Our results indicate that there was not a perfect correlation between the number of identified glands and post-operative parathormone in a subset of patients. PMID- 19902850 TI - Effects of different suture materials on wound healing and infection in subcutaneous closure techniques. AB - BACKGROUND: In this study we aimed to investigate the effects of different suture materials on wound healing and infection in subcutaneous closure techniques. MATERIALS AND METHODS: Twenty-nine rats were randomly allocated to 5 groups. All subjects received 2-cm vertical dermal and subdermal incisions on their backs under sterile and proper surgical conditions. The subdermal cuts were then closed with vicryl (polyglycolic acid), biosyn (monofilament glycomer), prolene (polypropylene) and tissue adhesive (2-octylcyanoacrylate). After 10 days, all sutured areas were examined for seroma, haematoma formation and cosmetic outcome. The incisional lines were excised with 1-cm-wide skin strips and tissue hydroxyproline levels were determined. The specimens obtained were evaluated for the presence of giant cells, mononuclear cells, fibrosis and neutrophils. RESULTS: No differences in tissue hydroxyproline levels were found between any of the suture materials used. Fibrotic process and inflammatory cell infiltration were more prominent in the biosyn and prolene groups than in other groups. Foreign body giant cells were observed in the biosyn group. CONCLUSION: Vicryl and tissue adhesive should be preferred in subcutaneous closure techniques. PMID- 19902851 TI - Paranasal sinus complications caused by dental implants and complementary procedures. AB - OBJECTIVE: To describe the paranasal complications that may occur after dental implantation. STUDY DESIGN: A literature search was performed in order to review currently available information about the complications of dental implantations and complementary procedures. RESULTS: The use of dental implants has gained great popularity due to their convenience, natural look, and better speech and chewing efficiency, as well as their potential to slow and even stop jawbone atrophy. Nevertheless, an increasing number of publications have drawn attention to the possible complications of the procedure for the paranasal sinuses. Both short-term and long-term complications have been described that should be familiar to otolaryngologists. CONCLUSIONS: Although the procedure is performed by dentists, dental implantations may also have a major impact on the paranasal sinuses. Complications may necessitate the knowledge and cooperation of both dentists and otolaryngologists. PMID- 19902852 TI - Increased frequency of rhinitis medicamentosa due to media advertising for nasal topical decongestants. AB - AIM: The aim of this study was to investigate the frequency of rhinitis medicamentosa (RM) in patients attending the ENT outpatient clinic of the General Hospital of Rethymnon (Crete, Greece) before and after the launch of an intensive nasal decongestant advertising campaign in Greece. METHODS: We reviewed the medical records of the patients with RM seen at the ENT outpatient clinic in May, June and July of 2003 and 2006. We analyzed and recorded the gender, age, and related clinical information of the patients with RM. RESULTS: In May, June and July of 2003, 41 patients out of a total of 1780 patients attending the ENT outpatient clinic were diagnosed with RM (2.3%). In the same months in 2006, 161 patients out a total of 1898 patients were diagnosed with RM (8.5%). The frequency of RM in these groups was therefore found to have increased significantly between 2003 and 2006. In 2006, 8 out of 10 patients with RM reported that they had made their purchasing decision solely on the basis of the information supplied by the drug advertisement without consulting their doctor or pharmacist. CONCLUSION: We suggest that the intensive media advertising campaign for nasal topical decongestants (particularly on TV) which started in 2004 is probably the main reason for this "endemic" RM. PMID- 19902853 TI - Management of post-infectious olfactory dysfunction: a nationwide survey of UK ENT consultants. AB - OBJECTIVE: To determine the current practice of UK ENT consultants in investigating cases of suspected post-infectious olfactory dysfunction. METHOD: We prepared questionnaires and posted these to every consultant ENT surgeon registered with the British Association of Otolaryngologists Head and Neck Surgeons (BAO-HNS). Questions included preferred radiological investigation, smelling test and follow up. RESULTS: A total of 510 questionnaires were sent, with a response rate of 51%. Most UK consultant ENT surgeons investigated suspected post-infectious olfactory dysfunction by performing either a CT (44%) or an MRI (29%) scan, but only 37% performed formal smell tests. Most consultant ENT surgeons (76%) decide not to follow up post-infectious olfactory dysfunction. CONCLUSION: This study gives an indication that suspected post-infectious olfactory dysfunction is not investigated thoroughly in the United Kingdom, with only a minority of clinicians performing formal smell tests and a lack of consensus as to radiological investigation. Standardisation of investigations is key to adequately managing suspected post-infectious olfactory dysfunction. PMID- 19902854 TI - Extramedullary plasmacytoma of the septum. AB - PROBLEM: Extramedullary plasmacytoma of the head and neck is a rare neoplasm characterized by monoclonal proliferation of plasma cells. The nasal cavity and nasal septum are the most common sites of occurrence and the neoplasm can be solitary or multiple. Extramedullary plasmacytoma is associated with the initial appearance of multiple myeloma and may precede overt manifestations of systemic disease by months or years. METHODOLOGY: A seventy-year-old female presented to our clinic with a one-month history of nasal obstruction. We performed a systematic approach to diagnosis using clinical, laboratory, and radiologic investigations in order to exclude systemic involvement. RESULTS: The patient was diagnosed with extramedullary plasmacytoma and the mass was excised completely via transnasal endoscopy. CONCLUSION: Extramedullary plasmacytoma of the nasal cavity is rare and should be considered in the differential diagnosis of nasal cavity masses. PMID- 19902855 TI - Lethal sphenoid fungal sinusitis in an immunocompromised elderly patient. AB - Sphenoid fungal balls are rare and typically indolent lesions, unless superinfected by bacteria or invasive to adjacent neurovascular structures. If the identification or treatment of underlying complications is delayed in an immunocompromised patient, a catastrophic outcome may result. We report the case of an elderly female patient with poorly controlled diabetes mellitus suffering from sphenoid fungal sinusitis that is complicated by cavernous sinus thrombophlebitis and carotid artery stenosis. In spite of reasonable diagnosis and therapy, the patient's general condition deteriorated and she eventually died. The clinical presentation, diagnosis, and treatment strategy are discussed. PMID- 19902856 TI - Endonasal endoscopic dacryocystorhinostomy for a primary dacryocystocele in an adult. AB - A dacryocystocele refers to a sterile cystic dilatation of the lacrimal sac resulting from a complete obstruction of the lacrimal pathway above and below the lacrimal sac. In most cases, it is a congenital disorder that typically occurs in neonates. It is characterized by a bluish cystic swelling just below the medial canthus accompanied by epiphora. In some cases, there is also an intranasal expansion beneath the inferior turbinate. When this expansion is large and bilateral, it can lead to respiratory distress in neonates since they are obligate nasal breathers. Although congenital dacryocystoceles may resolve with conservative measures, many become infected and require systemic antibiotic treatment, and most require early surgical intervention. Herein, the authors report a rare case of dacryocystocele in an adult that was successfully managed with an endonasal endoscopic dacryocystorhinostomy. The pre-operative images and the intra-operative findings are shown, and the available literature is reviewed. PMID- 19902857 TI - An unusual cause of dysphonia: laryngeal verruca vulgaris. AB - PROBLEMS/OBJECTIVES: Verruca vulgaris of the larynx (VVL) is an uncommon lesion with clinical and histopathological features that are easily misdiagnosed as verrucous carcinoma, and thus may lead to unnecessary surgery. This case study investigated clinical and pathological features that might distinguish VVL from other keratotic laryngeal lesions. METHODOLOGY: A 35-year-old man presented with a 3-month history of progressive hoarseness and superficial, keratotic, vocal cord lesions. The verrucoid configuration of the surface and lack of nuclear atypia enabled treatment by local excision. The biopsied lesion was sectioned and examined by immunohistochemistry, and in situ hybridization. RESULTS: The patient was disease-free up to 1 year and 6 months after surgery. The exophytic, keratotic lesions of the larynx had features of cutaneous verruca vulgaris. No human papilloma virus was detected. CONCLUSIONS: We described histopathological features of VVL that will facilitate its distinction from laryngeal keratosis, squamous papillomas, verrucous hyperplasia, and verrucous carcinoma. PMID- 19902858 TI - Castleman's disease of the neck: report of a case and review of the literature. AB - OBJECTIVES: Castleman's disease is an uncommon disease of benign lymph node hyperplasia primarily affecting the mediastinum, with the head and neck region being the second most common site. CASE REPORT: A 27-year-old woman was admitted to our department due to a left lateral cervical mass. After a complete clinical and imaging examination, a neck dissection was performed, in which a mass, size 3.5 x 2.7 x 4.5 cm, was excised. The patient was diagnosed with Castleman's disease of the neck after histopathological examination. CONCLUSIONS: The diagnosis of Castleman's disease is always a clinical challenge, as the patient commonly presents with nonspecific signs and symptoms resembling other lymphatic diseases. Consequently, Castleman's disease should be in the differential diagnosis of congenital, inflammatory, or neoplastic cervical alterations. A review of literature, including histopathological characteristics, differential diagnosis, and treatment options is also presented. PMID- 19902859 TI - A historical vignette (15). "Be proud of yourself: you have a history!" The nose and the plague. AB - The nose and the plague. Although the plague does not cause any specific nasal pathology, the miasma theory and the repulsive smell of the disease were factors that contributed to a strong emphasis on the nose. To stop the spread of the disease, it was thought necessary to saturate the nose with protective scents (hence the nose of the plague doctors) (Figure 1), or simply to hold one's nose. Moreover, the nose was long considered to be an outlet for mucus from the encephalon, and so induced nose bleeding and sneezing were advised when the plague seemed to be attacking the brain. PMID- 19902860 TI - Octreotide in chemotherapy induced diarrhoea in colorectal cancer: a review article. AB - BACKGROUND: Chemotherapy-induced diarrhoea (CID) is well known in cancer management. The risk is greater when the primary cancer is colorectal. This article aims towards assessing the role of octreotide in CID through an extensive literature search. METHODS: After searching through PUBMED, MEDLINE and the Cochrane library, only those studies which were published over the last 20 years in English and where at least the majority of the cohort were colorectal patients, were included. Two randomized trials, four non-randomized studies and two case-series publications were thus considered. RESULTS: It was seen in both the randomized studies, that octreotide had much better outcome as compared to loperamide in treating severe CID. Among 88 patients from the non-randomized studies with severe CID, the primary cancer was colorectal in 79 patients. 61 patients had drug-resistant CID. Within a maximum of 96 hours, octreotide reduced CID by > or = 2 grades in 91% of 88 patients and in 88.52% patients with drug resistant CID. CONCLUSION: Octreotide is effective in treating severe CID, resistant to other modes of treatment. It is associated with a few minor adverse effects. Though expensive, octreotide could be considered as first line medication in CID of grades 3 or above. Its use in lower grades of CID would not be cost effective. PMID- 19902861 TI - High fat consumption in children with celiac disease. AB - AIM: The purpose of this study was to estimate the caloric intake and fat consumption in children with celiac disease (CD) following a gluten-free diet (GFD). PATIENTS AND METHODS: This study enrolled 100 subjects, including 50 children with CD on a gluten-free diet and a control group of 50 healthy children. Statistical analysis to compare groups was performed using one-way ANOVA. RESULTS: A significant increase in fat consumption was observed in children with CD as compared to healthy children. The daily fat intake was 72.5 +/- 37.2 g per 100 g of food in the CD group and 52.9 +/- 35.4 g per 100 g of food in the control group (p < 0.008). A significant difference in fat intake was found between celiac and healthy females (10.21 +/- 3.15 g/100 g in the celiac group vs 7.46 +/- 2.91 g/100 g in the control group), p = 0.004. CONCLUSIONS: This study describes a significantly higher fat consumption in patients with CD on GFD as compared to controls. This increase was more pronounced in females and during the puberal age. Based on these interesting preliminary results we estimate that further investigations are necessary, such as a randomized multicentre study on the long-term effects of GFD with particular attention to the imbalance in daily fat intake. PMID- 19902863 TI - An anti-preS2 antibody protects human hepatocytes from hepatitis B virus infection. AB - BACKGROUND: Hepatitis B virus infection is a major problem in liver transplantation. In this study, we examined the potential efficay of a recombinant adenovirus expressing an antibody against the HBV preS2 antigen (Ab-H HBV-S2) in preventing HBV recurrent infection after liver transplantation. METHODS: A gene for humanized antibody against the HBV preS2 antigen was cloned into pDC315, a type 5 adenoviral shuttle plasmid. Recombinant virus was obtained by homologous recombination in the 293 packaging cells. The virus containing the Ab-H-HBV-S2 gene was transduced into the rat liver graft during cold preservation. The recombinant virus produced antibody and showed protective effects on human hepatocytes from hepatitis B virus infection in vitro. RESULTS: The recombinant virus titer determined by TCID50 analysis was 5.1x10(10) PFU/mL. The concentration of preS2 antibody in BALB/C nude mice was 16.7 +/- 10.5 microg/mL on day 3,30.9 +/- 13.6 pg/mL on day 7, and lasted for 5 weeks after the injection. At a concentration of 0.5 microg/mL or above, the preS2 antibody protected cultured human hepatocytes from hepatitis B virus. CONCLUSIONS: Adenovirus-mediated gene transduction of anti-preS2 antibody in the transplanted liver may be a useful approach to prevent hepatitis B infection after liver transplantation. PMID- 19902862 TI - Diagnostic value of antineutrophil cytoplasmic antibodies and anti-Saccharomyces cerevisiae antibody in Iranian patients with inflammatory bowel disease. AB - BACKGROUND AND STUDY AIMS: Perinuclear antineutrophil cytoplasmic autoantibodies (pANCA) and anti-Saccharomyces Cerevisiae antibody (ASCA) are potential markers for diagnosis of inflammatory bowel disease (IBD). The aim of the present study was to evaluate the diagnostic value of pANCA and ASCA in Iranian patients with IBD. PATIENTS AND METHODS: Serum samples were collected from 144 patients with IBD (113 ulcerative colitis and 31 Crohn's disease) and patients with non-IBD problems were assayed for ASCA by Enzyme-Linked Immunosorbent Assay (ELISA) and for pANCA by indirect immunofluorescence assay. RESULTS: Sensitivity and specificity of pANCA in UC were 39.8% and 82.1%, respectively. For CD, pASCA test provided the sensitivity of 58% and specificity of 70%. A combination of pANCA+/ASCA- for diagnosis of UC showed a sensitivity of 31.9% and specificity of 89.1%. In addition the combination of pANCA-/ASCA+ showed a sensitivity of 35.5% and specificity of 79.8% for diagnosis of CD. CONCLUSION: Due to low sensitivity of pANCA and ASCA alone or in combination, they are not valuable serological markers for diagnosis of UC or CD. PMID- 19902864 TI - Physiologic variables for videofluoromanometric assessment of dysphagia: an exploratory study. AB - STUDY AIM: To assess the physiological variables among Upper Esophageal Sphincter Nadir (UESN), Hypopharyngeal Peak Pressure (HPP) and Pharyngo-Esophageal Pressure Gradient (PEPG) for Videofluoromanometry (VFM). PATIENTS & METHOD: Exploratory non-randomised prospective study comparing UESN, HPP and PEPG of three cohorts of individuals presumably presenting very distinctive "manometric signatures" based on McConnel's Piston Model of swallowing: 11 non-dysphagic volunteers called the Control Group (CG), 10 dysphagic patients presenting a Myotonic Dystrophy (MD), at various stages of evolution, and 10 patients presenting a CricoPharyngeal Barr (CPB), with no post-swallow pharyngeal residue at a previous Modified Barium Swallow (MBS). VFM tests are performed using solid-state three unidirectional transducers produced by Gaeltec Inc. The simultaneous display storage of the standard fluoroscopic swallow of 10 ml liquid barium with UESN and HPP measurements, continuously recorded on a 3-channel polygraph, is performed using a Kay-Pentax Swallowing Work Station. PEPG calculations are subsequently made. RESULTS: Significant different HPP and PEPG values were observed between the three cohorts. MD patients presented HPP and PEPG below 100 mmHg while CPB patients presented HPP and PEPG above 200 mmHg. The CG presented HPP and PEPG between 100 and 200 mmHg. UESN values revealed no significant difference between the three cohorts. A reading scale is proposed. The aim of the scale is to make a link between HPP or PEPG values and physiopathological (not diagnostic) conditions. Patients presenting an HPP or PEPG below 100 mmHg indicate a High probability of Pharyngeal Propulsion Impairment while patients presenting an HPP or PEPG above 200 mmHg are more likely to have an Increased Flow Resistance with appropriate Propulsion Response. Pros and cons for calculation of the PEPG, representing a possibly unnecessary step, are discussed. CONCLUSIONS: In our study, the use of HPP or PEPG as physiological variables provides quantitative data that allow VFM to discriminate three very distinctive swallowing conditions. Further studies are needed to assess the HPP and PEPG obtained with other manometic devices within the same specific populations for them to be considered as universal physiological variables. Eventually, further investigations should answer the question as to whether the calculation of the PEPG represents any value in comparison with HPP measurement alone. PMID- 19902865 TI - Surgical management of hepatic metastases of colorectal origin. AB - Colorectal cancer is the most frequent digestive cancer. Prognosis is greatly depending on the TNM stage at the time of diagnosis. Fifty percent of all patients shall develop, synchronously or metachronously, liver metastases. Different means such as chemotherapy, targeted therapies, radiofrequency ablation, portal vein embolization and two-stage hepatectomy may be used to make these metastases eventually resectable and to increase overall survival. This is a short review of these different methods used to increase resectability but also on the integration of these parameters in a larger approach of colorectal liver metastasis surgery especially insisting on multidisciplinary discussion. PMID- 19902866 TI - Osteonecrosis in inflammatory bowel diseases: a review of the literature. AB - INTRODUCTION: Osteonecrosis (ON) of the femoral head can lead to femoral head collapse, necessitating total hip replacement. Reports of patients suffering from both ON and Inflammatory Bowel Diseases (IBD) have prompted us to evaluate the relationship between ON and IBD, especially Crohn's disease and ulcerative colitis. METHODS: A review of the data from three new cases, along with data from all the published cases of patients presenting ON and IBD found through a systematic search of the Pub Med database. RESULTS: We encountered some diagnostic problems: The ON diagnosis could not be confirmed in some patients who did not meet the ON diagnostic criteria. Reviewed data was too weak to assess the exact incidence of ON in IBD. Corticosteroid therapy, especially in high dose regimens, is likely the most important etiological factor. No evidence supporting other physiopathological hypothesis, such as total parenteral nutrition, osteoporosis, or coagulation disorders, was found. Finally, the multifocal form of ON appears particularly common in IBD, with some patients presenting multiple lesions of the hip, shoulder, knee and talus. CONCLUSIONS: ON in IBD, which is frequently multifocal, appears to be a complication of corticosteroid therapy, especially when high doses are used. We recommend regular ON checkups for corticosteroid-treated IBD patients. PMID- 19902867 TI - History of endoscopic devices for the exploration of the small bowel. AB - The small bowel has gained new attention since the development of the wireless videocapsule in 2000, opening up the last 'black box' of the gastrointestinal tract. Although conventional push enteroscopy has been available for decades, since the wireless videocapsule new enteroscopes have been developed to examine the entire small bowel endoscopically in order to perform all conventional endoscopic procedures. The present review highlights the historic evolution of enteroscopy, from the first complete enteroscopy in 1971 over the current balloon assisted and over-tube-guided methods of enteroscopy to future directions of evolutions towards perfection. PMID- 19902868 TI - The dendritic therapy with its potential applications in pancreatic cancer. PMID- 19902869 TI - The Maastricht III consensus: summary and comments. AB - The management of Helicobacter pylori infection as been readapted to recent information which resulted in new guidelines that were published in 2007 in Gut. Iron deficiency anemia with negative work up and idiopathic thrombocytopenic purpura were added to the indications for treatment while the known ones were reconfirmed. Much interest went to prevention of gastric cancer and treatment. In the prevention of gastric cancer the importance of treating infected individuals before the appearance of premalignant lesions was highlighted. The most important problem concerning treatment is the increasing resistance for clarithromycin with as a consequence a decreasing efficiency of the classical therapy. Alternatives are discussed and results of resistance for different antibiotics in Belgium are presented. PMID- 19902870 TI - Cerebral venous sinus thrombosis as presenting feature of ulcerative colitis. AB - Thrombosis is a well recognized complication of inflammatory bowel disease that occurs in 1.3 to 6.4% of patients, however, cerebral vascular involvement is unusual. We present the case of a 16-year-old female in whom cerebral venous thrombosis was the presenting symptom of an active ulcerative pancolitis. Thrombophilia screen (plasma levels of proteins C and S, antithrombin, antibeta2 glycoprotein, lupus anticoagulant and anticardiolipin antibodies, activated protein C resistance, homocystein level antinuclear antibodies) was negative. The patient was successfully treated with anticoagulant therapy, phenobarbital and sulfasalazine. Cerebral venous thrombosis is an exceptional presenting feature of ulcerative colitis. Disease activity may play a major role in the occurrence of thrombosis. PMID- 19902871 TI - Emphysematous gastritis causing gastric and esophageal necrosis in a young boy. AB - Emphysematous gastritis is a rapidly fatal and rare type of infectious gastritis. It may lead to involvement of esophagus, and organ necrosis, in its severe form. A 16-year-old, previously healthy, boy presenting with acute abdomen was diagnosed to have emphysematous gastritis on CT scan. During laparotomy, there was complete necrosis of the stomach, with patchy esophageal involvement. Aggressive management in the form of total gastrectomy, and later, transthoracic esophagectomy was done. However, it failed to alter the course of the illness, and the patient succumbed to the illness. Emphysematous gastritis is rare in young patients without known risk factors. Also, only two previous cases have been reported with esophageal involvement. We have presented this case with a brief review of literature. PMID- 19902872 TI - Endoscopic closure of a large iatrogenic rectal perforation using endoloop/clips technique. AB - Retroflexion to evaluate the rectal vault provides significant additional information compared with standard forward view of the rectum. The procedure is easily performed with rare complications and is well tolerated by patients. We describe the first case of a large oval rectal perforation after retroflexion of the colonoscope in a healthy rectum during a follow-up colonoscopy, immediately closed with the endoloop/clips technique. The patient had an uneventful course and was discharged after 5 days. At his 2-month follow-up visit he remained asymptomatic and endoscopy revealed complete healing of the perforation. PMID- 19902873 TI - Repair of an EUS--induced duodenal perforation with endoscopic clips. AB - Endoscopic ultrasound (EUS) is considered a safe procedure; however, rare deaths have been reported due to complications such as perforation of the gastrointestinal tract. Several factors including age, the presence or absence of cervical osteophytes or duodenal diverticula, history of difficult intubation with prior endoscopic procedure, endosonographer's inexperience, or EUS guided interventions such as the drainage of the pancreatic duct or pseudocyst and fine needle aspiration may increase the risk of EUS related perforation of the gastrointestinal tract. We report a patient with pancreatic mass who developed duodenal perforation during EUS and was treated successfully with an immediate closure of perforation using endoscopic clips combined with bowel rest and antibiotics. Based on our patient and others reported in the literature, immediate recognition and closure of perforation with endoscopic clips may be useful in the management of patients with EUS induced duodenal perforation. However, surgical consultation and close clinical monitoring is required in the management of these patients. PMID- 19902874 TI - Abdominal pain and vomiting as first sign of mitochondrial disease. AB - We describe a patient in whom abdominal pain and vomiting were the presenting symptoms of Mitochondrial Myopathy Encephalopathy, Lactic Acidosis with Stroke like episodes syndrome (MELAS). Mitochondrial disorders usually present with neurological symptoms or with myopathic features at any age. Although many patients develop visceral symptoms at a certain moment during the course of the disease, only in a minority of patients these symptoms are the unique presenting ones. The proband was initially diagnosed as having gastro-oesophageal reflux and it was only after detailed clinical history that an underlying metabolic defect was suspected and the molecular defect identified. PMID- 19902875 TI - Primary myeloid sarcoma of the jejunum and greater omentum causing small intestine obstruction. AB - Myeloid sarcoma, which is highly associated with acute myeloid leukemia, is defined as an extramedullary discrete tumor mass, consisted by immature myeloid cells or myeloblasts. Myeloid sarcoma usually involves the skin, lymph node, bone, soft tissue and testis, while involvement of the gastrointestinal tract is rather uncommon. The diagnosis depends on histological features and immunohistochemical results. We present a rare case of myeloid sarcoma, with synchronous involvement of the jejunum and the greater omentum, manifesting with small bowel obstruction. PMID- 19902876 TI - Esophageal squamous papillomatosis with dysplasia. Is there a role of balloon based radiofrequency ablation therapy? AB - Esophageal squamous papillomatosis (ESP) is a rare condition, occurring in an estimated 0.01-0.097% in data from upper gastrointestinal endoscopies and autopsy series. Chronic mucosal irritation and infection with human papilloma virus (HPV) are proposed etiologies. Heavy use of tobacco and alcohol are common associations. The premalignant potential of ESP has long been debated in literature. The clinical course is variable, ranging from spontaneous regression to the development of squamous cell carcinoma. Due to the paucity of reported cases, no generalized therapeutic or surveillance recommendations exist. Photodynamic therapy (PDT) has been successfully used to treat Barrett's esophagus as well as superficial adenocarcinoma. However, its safety and efficacy in treating ESP with dysplasia is lacking. Balloon-based radiofrequency ablation using the HALO90 Ablation System is designed to remove the diseased cells using controlled heat. In recent clinical trials, it has shown great promise in treating Barrett's esophagus with high-grade dysplasia. We report the first ever use of balloon-based radiofrequency ablation to treat ESP with dysplasia. Clinical symptoms resolved after the first therapy session, however, ablation therapy was terminated early because squamous cell carcinoma in-situ was detected on surveillance endoscopy prior to the fourth therapy session. Although we failed to treat type 4 ESP with high-grade dysplasia with balloon-based radiofrequency ablation therapy, we believe that it might play a role in treating other localized types of ESP. PMID- 19902877 TI - Hepatocellular carcinoma in an adult patient with type IV glycogen storage disease. PMID- 19902878 TI - Abdominal wall scar endometriosis. A report of two cases with emphasis on diagnostic strategies. PMID- 19902879 TI - Surgical treatment of an acetabular fracture and labral tear with suture anchors in a 10-year-old child. AB - This article presents a case of a 10-year-old boy with a right acetabular fracture secondary to a direct blow to the pelvis and back. He sustained a right hip dislocation and transverse with posterior wall acetabular fracture with a nonconcentric hip joint after reduction. The fracture was treated with right labral and articular segment absorbable suture tie-down and relocation of the right hip into an anatomical position. At 32-month follow-up, the patient had returned to activities of daily living, he had a normal and symmetric gait pattern, and no pain on hip adduction, flexion, or internal rotation. This technique reduced the fracture and labrum, relocated the hip joint, and obviated the need for hardware removal that is typically needed in the pediatric population. This case is distinctive due to the patient's young age and treatment method, which has not been described previously for the treatment of this rare pediatric fracture. PMID- 19902880 TI - Acetabular cup malalignment after total hip resurfacing arthroplasty: a case for elective revision? AB - This article describes the clinical course of a patient with a resurfacing implant in a poor cup position in combination with elevated serum metal ions prior to implant failure. Following resurfacing, the patient had substantial improvement from baseline in pain and functional status. Postoperative radiographs indicated the acetabular cup in an abducted and excessively anteverted position. The acetabular component ultimately failed after 4.5 years and a traditional total hip arthroplasty revision was performed. Serum cobalt (Co) and chromium (Cr) concentrations had been collected postoperatively of the index procedure at 6 months, 1 year, 2 years, 3 years, and pre- and postoperatively at the time of implant revision. Serum cobalt and chromium ion levels were progressively elevated to approximately 400 times more than the expected range at all time points prior to revision. Elective revision had been considered due to acetabular malalignment and elevated metal ion levels, but not performed since the patient was doing well clinically. A recent study has shown a correlation between increased cup inclination and increased serum cobalt or chromium levels and this patient's levels were >40 times greater than that typically observed with this device. Early revision should be strongly considered if component malpositioning is noted, and abnormally elevated ion concentrations should signal the need for revision regardless of the patient's clinical status. The relationship of a malpositioned cup and uncharacteristically elevated metal ion levels is related to the metal-on-metal bearing coupling and likely applies to conventional metal-on-metal total hip prostheses as well. PMID- 19902881 TI - Spontaneous disappearance of lumbar disk herniation within 3 months. AB - Although spontaneous regression of disk herniation is a well-known phenomenon, the time taken for the condition to resolve has not been detailed in previous studies. This article describes a case of vanishing lumbar disk herniation in a 33-year-old man. The patient experienced sudden severe lumbar pain while lifting a 240-kg weight while attempting a Donkey Calf Raise during muscle training. The pain persisted despite the use of a lumbar corset and nonsteroidal anti inflammatory drugs. Twelve days after onset, sensory disturbance appeared in the right L5 dermatome, and a manual muscle test of the right anterior tibial muscle revealed level 3. The pain gradually spread over the right lower extremity and the indistinct lumbago changed to localized back pain at the L4/5 vertebral level. Magnetic resonance imaging (MRI) of the lumbar spine 12 days after onset revealed a large disk herniation at L4/5. T2-weighted images demonstrated the herniated disk with a sequestrated disk fragment, which compressed the right L5 nerve root. Over the following month, his pain gradually diminished and he was able to resume his muscle-building program. Follow-up MRI 3 months after the lumbar injury showed complete disappearance of the extruded disk material. This is the first reported case of disk herniation that disappeared within only 3 months, as previous reports have reported that a minimum 30-week period was needed. Clinical awareness of the possibility that disk herniation may resolve within a relatively short time may aid both correct informed consent and treatment. PMID- 19902882 TI - Anatomic anterior cruciate ligament reconstruction in the skeletally immature: is it possible? PMID- 19902883 TI - Malignant hyperthermia: a pharmacogenetic disorder. PMID- 19902884 TI - ACL reconstruction in adolescent and preadolescent patients. PMID- 19902885 TI - Vacuum-assisted closure with external fixation of the hand. PMID- 19902886 TI - Biologic response to orthopedic sutures: a histologic study in a rabbit model. AB - Biologic reactivity to suture materials can have an effect on patient outcomes. The goal of this study was to determine the histologic response to 8 commonly used orthopedic sutures--Ethibond (Ethicon, Somerville, New Jersey), Ticron (Tyco, Waltham, Massachusetts), HiFi (Linvatec, Largo, Florida), Ultrabraid (Smith & Nephew, Memphis, Tennessee), MaxBraid (Biomet, Warsaw, Indiana), Orthocord (Mitek, Raynham, Massachusetts), MagnumWire (Opus Medical, San Juan Capistrano, California), and FiberWire (Arthrex, Naples, Florida)--using a rabbit model. The suture granuloms were evaluated at 30, 60, and 120 days with measurement of the fibrous capsule, the number of giant cells in and near the capsule, and the overall inflammatory grade: 1 (mild), 2 (moderate), and 3 (severe). MagnumWire and Ticron sutures initiated a more intense inflammatory reaction when compared to the other sutures. By 120 days, MagnumWire (P=.0297) and Ticron (P=.1855) had fewer giant cells at the soft tissue-suture interface, fewer giant cells within the capsule (P=.0074 and P=.0377, respectively), and the greatest capsular thickness of all suture types (P<.0001 and P=.1378, respectively). Differences exist between the biologic reactivity of commonly used orthopedic sutures that may be attributable to their material composition and/or braid characteristics. In comparison to other high-strength sutures, MagnumWire and Ticron stimulated a more intense foreign body inflammatory response. PMID- 19902887 TI - The effects of postoperative ketorolac on wound healing in a rat model. AB - Ketorolac is a nonopioid, anti-inflammatory drug commonly used for postoperative analgesia. Its effectiveness has been previously documented in various orthopedic procedures and more recently in spinal surgery. It remains uncertain if ketorolac has an effect on wound healing. The purpose of this study was to determine if the use of postoperative ketorolac induced deleterious effects on wound healing in a simulated spinal surgery incision using a rat model. A 4-cm dorsal midline incision was made and closed in 36 rats. Rats were divided into 3 groups: (1) 5 mg/kg ketorolac given every 6 hours for 24 hours; (2) 5 mg/kg ketorolac given every 6 hours for 48 hours; and (3) control group given dextrose 5% in water every 6 hours for 48 hours. On postoperative day 14, sutures were removed. Wounds were removed and loaded to failure in tension. The mean+/-SD loads to failure were 9.8+/-1.8 N for group 1, 9.0+/-2.4 N for group 2, and 9.5+/-4.5 N for group 3. The differences among the 3 groups were not statistically significant (P>.05). The use of ketorolac in the immediate postoperative period produces no increased risk of wound complications in this rat model. PMID- 19902888 TI - Membrane type-1 metalloproteinase mediates nitric oxide-induced activation of matrix metalloproteinase-13. AB - This article investigates the mechanism of activation of matrix metalloproteinase 13 induced by nitric oxide. SW1353 cells were stimulated with S-nitroso-N-acetyl D,L-penicillamine, expressions and activities of metalloproteinase-13, and membrane type-1 metalloproteinase were assayed, and a proteolytic activation of recombinant human metalloproteinase-13 was measured in the presence of recombinant human membrane type-1 metalloproteinase. Nitric oxide increased expressions of both matrix metalloproteinases and stimulated the proteolytic processing of metalloproteinase-13 from the pro-enzyme to the final active form. Recombinant human membrane type-1 metalloproteinase was able to process recombinant human metalloproteinase-13 to fully active enzyme. S-nitroso-N-acetyl D,L-penicillamine had no effect on recombinant human metalloproteinase-13 activity. Nitric oxide scavenger (OxyHb) strongly attenuated nitric oxide-induced proteolytic activation of metalloproteinase-13. Furthermore, tissue inhibitor of metalloproteinase-2 markedly reduced the activation of metalloproteinase-13 in response to nitric oxide. These studies identify membrane type-1 metalloproteinase as a target for nitric oxide, which may be critical for the nitric oxide-induced activation of metalloproteinase-13. PMID- 19902889 TI - Noise characteristics of surgical space suits. AB - Several studies indicate that the noise generated by performing orthopedic surgery has the potential to cause hearing loss. Noise-induced hearing loss was found in 50% of the orthopedic surgeons studied using audiometric testing, with a greater incidence associated with years of practice. Noise produced by several orthopedic surgical instruments such as saws, drills, and hammers during surgery exceeds 100 dB, especially during knee replacement procedures. In one study, surgical space suits (personal protection systems) were suggested to help protect against noise-induced hearing loss, although space suit manufacturers do not market them as noise-reduction devices. A research protocol was developed to determine if commercially available surgical space suits help to reduce noise at the surgeon's ear. With the commercially available personal protection systems used in this research, there was no significant extra-helmet noise decrease by wearing the space suit. Sound inside the helmet at the level of the ear averaged 61 dBA, approximately the level of conversational speech, which may explain the difficulty the surgical staff may have hearing speech in the operating room when the space suit is worn with the fan on. If surgical noise is to be decreased, earplugs or muffs must be worn not only by the surgeon, but also by all personnel in the operating theater. At greatest risk may be the anesthesiologist, who may experience several orthopedic surgeries in a single day and is positioned close to the patient. PMID- 19902890 TI - Multivariate analysis of prognostic factors in synovial sarcoma. AB - Many studies have described the diversity of synovial sarcoma in terms of its biological characteristics and clinical features. Moreover, much effort has been expended on the identification of prognostic factors because of unpredictable behaviors of synovial sarcomas. However, with the exception of tumor size, published results have been inconsistent. We attempted to identify independent risk factors using survival analysis. Forty-one consecutive patients with synovial sarcoma were prospectively followed from January 1997 to March 2008. Overall and progression-free survival for age, sex, tumor size, tumor location, metastasis at presentation, histologic subtype, chemotherapy, radiation therapy, and resection margin were analyzed, and standard multivariate Cox proportional hazard regression analysis was used to evaluate potential prognostic factors. Tumor size (>5 cm), nonlimb-based tumors, metastasis at presentation, and a monophasic subtype were associated with poorer overall survival. Multivariate analysis showed metastasis at presentation and monophasic tumor subtype affected overall survival. For the progression-free survival, monophasic subtype was found to be only 1 prognostic factor. The study confirmed that histologic subtype is the single most important independent prognostic factors of synovial sarcoma regardless of tumor stage. PMID- 19902891 TI - Osteoid osteoma: a diagnosis for radicular pain of extremities. AB - Osteoid osteoma is a benign tumor commonly found in young patients. The most interesting feature of the tumor is its peculiar obvious pain. The intensity of pain produced by osteoid osteoma is unproportionate to its nature and size. Much has been written about the mechanism of this symptom of the tumor and the specific characteristics of the pain that can be misleading when appearing as radicular pain especially when some subtle neurologic signs and symptoms accompany the pain. In this article, 12 patients who were referred to our clinic with radicular pain and neurological signs and symptoms months after initiation of symptoms between 2000 and 2008 are presented. Of these 12 patients, most were first thought to have root compression disease before being referred to our hospital. Precise history taking and special attention to physical examinations led to a suspicion of osteoid osteoma. Plain radiographs, computed tomography, and bone scans indicated osteoid osteoma, and excisional biopsy confirmed its diagnosis. Osteoid osteoma should always be considered in young patients with radicular pain, even with subtle neurological signs and symptoms. PMID- 19902892 TI - Regeneration of the fibula using a periosteum-preserving technique in children. AB - Bone grafting is a commonly used surgical procedure, particularly in reconstructive orthopedic surgery. Bone grafts may be used to fill skeletal defects created by trauma, infection, tumors, or congenital malformation. Fibular graft is an ideal option and is widely used to treat massive segmental defects; however, it leads to various complications, especially in children. This article describes our experience with donor site bone grafts using a periosteum preserving technique with respect to minimizing donor site complications and performing effective reconstruction of bone defects in children. Seventeen patients who were followed after fibula resection and bone grafting in the donor site using a periosteum-preserving technique were evaluated radiologically and clinically. The patients were divided into 2 groups: 1 group was filled with cancellous allograft (n=9), and the other group was filled with calcium sulfate (n=8) in the donor site. Mean patient age was 8.4 years (range, 2-13 years), and mean follow-up was 31 months (range, 7-65 months). Complete regeneration of the donor sites occurred in all patients; mean time to regeneration at donor sites was 12 weeks (range, 4-21 weeks). There were no significant donor site complications intraoperatively or immediately postoperatively. The average regeneration index was 0.5 (range, 0.2-1.1), and it was found to increase with age (r=0.75, P<.001), indicating that younger patients had faster regeneration. The periosteum-preserving technique was found to lead to the reconstruction of bone defects effectively in children with minimal donor site complications. PMID- 19902893 TI - Arthrodiatasis for the treatment of Perthes' disease. AB - It is hypothesized that the interruption of the blood supply is an important factor causing femoral head osteonecrosis in the early stages of Legg-Calve Perthes disease. Currently, treatment by containment is recommended to direct and guide remodeling of the softened femoral head as it evolves from fragmentation through ossification. The goal of this study was to show the results of arthrodiatasis to induce height recovery of the femoral head and to achieve true ambulatory nonweight-bearing containment. Forty-two patients younger than 8 years with a diagnosis of Perthes' disease were studied. Twenty-three patients (9 class B and 14 class C according to Herring's classification) were treated with an articulated distraction technique and 19 patients (11 class B and 8 class C) were treated conservatively as a control group. Arthrodiatasis or articulated distraction of the hip combines off-loading of muscles and body forces with distraction of the joint space by means of an external fixator that crosses the hip joint. Radiologically, 21 patients (91%) had satisfactory results and 2 (9%) had unsatisfactory results. Clinically, the results were good in 21 patients (92%), fair in 1 (4%), and poor in 1 (4%). In patients treated conservatively, 14 patients (72%) had satisfactory results and 5 (28%) had unsatisfactory results. Clinically, 71% had good results, 17% had fair, and 12% had poor. We conclude that hip joint containment by articulated arthrodiatasis (plus adductors and psoas minimal tenotomy surgery) is an effective method in the management of Perthes' disease in patients younger than 8 years, classified B and C, and associated with a highly reduced range of abduction. Restoration of clinical abnormalities and satisfactory radiological parameters are achieved in high percentages. PMID- 19902894 TI - Biomechanical evaluation of 3-part proximal humerus fractures: a cadaveric study. AB - Proximal humerus fractures occur frequently, and treatment remains controversial. This study compares stiffness in axial loading for 3 methods of fixation (locking compression plate, standard (nonlocking) proximal humerus plate, and blade plate) in a 3-part proximal humerus fracture model. Twelve paired proximal humeri were obtained from embalmed human cadavers and separated into 3 groups. Osteotomies of the surgical neck and greater tuberosity were created to simulate a 3-part proximal humerus fracture. After fixation, constructs were axially loaded in 20 degrees of abduction for 200 cycles in an Instron materials testing machine (Norwood, Massachusetts). The blade plate (mean, 146.87+/-28.9 N/mm) demonstrated 29% more mean stiffness than the standard plate (mean, 113.0+/-22.3 N/mm; P=.19). The locking compression plate (mean, 130.71+/-39.2 N/mm) exhibited 15% greater stiffness compared to the standard plate in our 3-part model (P=.58). The blade plate demonstrated 12% greater stiffness than the locking compression plate (P=.64). There was no significant difference in mean stiffness between the fixed angle devices and the standard plate. Future in vitro and clinical studies of plate devices for proximal humerus fractures would be worthwhile to determine the benefits and limitations of various implants for specific types of fractures, including clinical performance and cost of care. PMID- 19902895 TI - Reproducibility and accuracy of templating uncemented THA with digital radiographic and digital TraumaCad templating software. AB - The reproducibility and accuracy of a digital software templating program on digital images was examined for primary uncemented total hip arthroplasty (THA). Forty-five patients waiting for an uncemented THA had templating performed of their digital picture archiving and communication systems (PACS) pelvic images with the TraumaCad software program (Orthocrat Ltd, Petach-Tikva, Israel). Acetabular cup size, femoral stem size, and femoral offset were noted by 2 observers, and again by 1 of the observers a week later. Through correlation coefficients and significance testing, the degree of intraobserver and interobserver variability was evaluated, as well as the level of accuracy for digital templating. Excellent correlation was found for all data sets, with no significant difference between them in intraobserver reproducibility. Also in terms of interobserver reproducibility, we found good levels of correlation, with no significant difference between the data sets. In terms of accuracy, the correlation was at least acceptable with no significant difference between any of the data sets. The use of the TraumaCad digital software program provides a reliable method of templating digital images for uncemented THAs. There is little intraobserver and interobserver variability, and the method produces an acceptable level of accuracy. PMID- 19902896 TI - The effects of anterior cruciate ligament lesion on the articular cartilage of growing goats. AB - Treatment of anterior cruciate ligament (ACL) injury in skeletally immature patients is controversial. The growth plate could be damaged if treated with the reconstruction techniques used to treat instability in adults. For this reason, many authors postpone surgical treatment until skeletal maturity, but the acceptable length of time that treatment can be postponed without causing irreversible damage to the articular cartilage in children with ACL injury is unknown. Until now, no studies have described the pathological findings and the evolution of the lesions of the articular cartilage during the growing period. For this reason, an experimental study on 16 6-month-old, skeletally immature goats was performed. A complete ACL lesion was achieved by removing the ligament. Two animals per group were sacrificed at intervals of 1, 3, 6, and 9 months postoperatively, and macroscopic and microscopic evaluations were performed. The presence of meniscal injury and articular cartilage lesions with progressive aspects were histologically underlined. The hystological observations showed that the complete ACL lesion causes irreversible articular cartilage alterations in growing goats 3 months after injury. These experimental data suggest that ACL reconstruction in growing patients with ACL injury and instability should be indicated without waiting until skeletal maturity. PMID- 19902897 TI - Range of flexion after primary TKA: the effect of soft tissue release and implant design. AB - Range of motion following total knee arthroplasty (TKA) is a crucial measure of clinical outcome. The purpose of this randomized, controlled study was to determine which factors are predictive of postoperative range of flexion. Fifty six patients received either a standard or a high-flexion design NexGen Legacy Posterior-Stabilized TKA (Zimmer, Warsaw, Indiana). The relationship between preoperative flexion, intraoperative flexion, and range of flexion 1 year postoperatively was determined. The influence of soft tissue release and the type of femoral component was also investigated. A significant correlation existed between preoperative flexion, intraoperative flexion, and maximum flexion 1 year postoperatively. Patients who had a preoperative range of flexion less than the mean range of flexion for the overall group gained flexion, whereas patients with a preoperative range of flexion greater than the mean range of flexion lost flexion. The degree of soft tissue release performed and the type of implant used had no influence on maximum flexion at 1 year. The principal predictive factor of postoperative range of flexion, regardless of the degree of soft tissue release or implant design, is the preoperative and intraoperative range of flexion. PMID- 19902898 TI - Surgical planning and procedures for difficult total knee arthroplasty. AB - Most total knee arthroplasties (TKAs) are performed for minimal knee joint deformities with standard techniques and instrumentation. However, patients with extra-articular deformities, severe varus and valgus deformities, and posttraumatic arthrosis pose unique challenges to surgeons. Each deformity requires different modifications of surgical technique or prosthesis used to successfully perform TKA and optimize postoperative results. The surrounding tissues connect the femur and the tibia at the anterior, posterior, medial, and lateral walls of the knee joint. The medial soft tissues should be released for varus deformities and lateral soft tissues should be released for valgus deformities. The posterior soft tissue may be released for flexed deformities. The anterior quadriceps and patellar tendon may be adjusted while approaching the knee joint for posttraumatic arthrosis. A more constrained knee prosthesis may be needed for more severe deformities. PMID- 19902899 TI - Accelerating autograft maturation in instrumented posterolateral lumbar spinal fusions without donor site morbidity. AB - Properly harvested iliac crest bone autograft applied to a meticulously prepared fusion bed produces a consistently high rate of fusion with a low incidence of donor site morbidity. Some reports advocate substituting bone morphogenic protein (BMP) for iliac crest bone autograft, but in posterolateral lumbar spinal fusion, BMP appears better suited to facilitate iliac crest bone autograft maturation than to substitute for it. In this single-center, nonrandomized, prospective study (minimum 2-year follow-up), cancellous-only iliac crest bone autograft was harvested for use in posterolateral lumbar spinal fusion. Reviewers blinded to graft condition and age assigned fusion scores to the random radiographs of 31 consecutive patients who underwent 1- to 3-level posterolateral lumbar spinal fusion using iliac crest bone autograft supplemented with either an implanted spinal fusion stimulator or BMP. There was no significant immediate or remote iliac crest bone autograft harvest morbidity, and there was a significant reduction in pain scores postoperatively (P<.001). At 12 months, BMP radiographs were more likely than spinal fusion stimulator radiographs to be rated as fused (P<.019). All BMP patients were deemed fused at 12 months and all spinal fusion stimulator patients at 24 months. In this study, iliac crest bone autograft supplemented with either BMP or spinal fusion stimulator resulted in a solid contiguous fusion without significant iliac crest bone autograft harvest-related morbidity. Bone morphogenic protein-supplemented iliac crest bone autograft fused at a faster rate, producing the more mature-appearing, trabeculated, robust fusion. PMID- 19902900 TI - Is the size appropriate for the procedure? PMID- 19902901 TI - Your diagnosis? Bipartite navicular. PMID- 19902902 TI - Thiophene/thieno[3,2-b]thiophene co-oligomers: fused-ring analogues of sexithiophene. AB - A series of six-ring oligothiophenes containing one to three degrees of ring fusion were assembled by a combination of metal-catalyzed Stille cross-coupling and oxidative homocoupling reactions. The effect of position and extent of ring fusion on the electronic properties was studied by UV-vis absorption and fluorescence spectroscopies, and these data were interpreted in the context of TD DFT computational analysis. Within each set of regioisomers, a slight red shift is revealed in the onset of the UV-vis absorption spectra when the fused-ring unit is located nearer to the periphery of the oligomer, indicating a narrower HOMO-LUMO gap. Incorporation of the unit of ring fusion toward the interior of the oligomer results in a decrease in the longest wavelength emission maximum and a reduced Stokes shift, and is accompanied by an increase in fluorescence quantum yield. PMID- 19902903 TI - Graphene oxide amplified electrogenerated chemiluminescence of quantum dots and its selective sensing for glutathione from thiol-containing compounds. AB - Here we report a graphene oxide amplified electrogenerated chemiluminescence (ECL) of quantum dots (QDs) platform and its efficient selective sensing for antioxidants. Graphene oxide facilitated the CdTe QDs*+ production and triggered O2*- generation. Then, a high yield of CdTe QDs* was formed due to the combination of CdTe QDs*+ and O2*-, leading to an approximately 5-fold ECL amplification. Glutathione is the most abundant cellular thiol-containing peptide, but its selective sensing is an intractable issue in analytical and biochemical communities because its detection is interfered with by some thiol containing compounds. This platform showed a detection limit of 8.3 microM (S/N = 3) for glutathione and a selective detection linear dependence from 24 to 214 microM in the presence of 120 muM cysteine and glutathione disulfide. This platform was also successfully used for real sample (eye drug containing glutathione) detection without any pretreatment with a wide linear range from 0.04 to 0.29 microg mL(-1). PMID- 19902904 TI - Guest-induced irreversible sliding in a flexible 2D rectangular grid with selective sorption characteristics. AB - A 2D noninterpenetrated flexible metal-organic porous solid, {[Cu(2)(cis chdc)(2)(bpee)] x H(2)O}(n) (1) based on a paddle-wheel building unit has been constructed using a mixed-ligand system. Guest-induced irreversible internetwork displacements of the 2D grids result in permanent porosity in the framework, as realized by the selective CO(2) sorption over N(2). The different affinity and selectivity of the solvent molecules was correlated with the internal polarity of the pore surfaces. PMID- 19902905 TI - Concise synthesis of pauciflorol F using a Larock annulation. AB - A Pd-catalyzed Larock annulation provides expedient access to a subset of resveratrol-derived natural products. The reported approach resulted in the structural revision of an intermediate en route to the natural product pauciflorol F, the total synthesis of which proceeded in two steps from the requisite pentannulation product. PMID- 19902906 TI - Copper(I), silver(I), and palladium(II) complexes of a thiaoxamacrocycle displaying unusual topologies. AB - Coordination behavior of the 14-membered dibenzo-O(2)S(2) macrocycle, L, with the soft metal ions such as Pd(II), Cu(I), and Ag(I) is reported. The X-ray structures of the complexes have been determined, and a range of less common structural types, including mono-, di-, and multinuclear species with discrete and continuous forms were obtained. A two-step approach via reaction of dichloro palladium(II) complex of L as a metalloligand, with L through a treatment of silver(I) perchlorate, led to the formation of bis(ligand) monopalladium(II) complex, [Pd(L)(2)] x 2 ClO(4) x 2 CH(3)NO(2) (1). In this case, the square planar stereochemical demand of Pd(II) coupled with the coordination of the S(2) donors of L to a metal center results in the formation of the bis(ligand)-type mononuclear complex. In 1, the ether oxygens of the ring are unbound. L reacts with CuBr to yield related 2:2:2 (metal/ligand/anion) complex of type [Cu(2)Br(2)(L)(2)] (2), in which two ligand molecules are linked by a rhomboid type Cu-Br(2)-Cu cluster unit. However, the parallel reaction with CuI afforded the mixture of the isostructural 2:2:2 type complex [Cu(2)I(2)(L)(2)] (3a) and double-stranded 1D coordination polymer {[Cu(2)I(2)(L)(2)] x 2 CH(3)CN}(n) (3b). The framework of 3b contains four bridging L coordinated to a rhomboidal Cu-I(2) Cu motif through the exomonodentate Cu-S bonds. Again, the ether oxygens do not coordinate in copper(I) halide complexes. The observed different reactivity between copper(I) bromide and copper(I) iodide was also confirm by an XRPD measurement. Unlike the Pd(II) and Cu(I) cases, L afforded an unusual tris(L) disilver(I) (club-sandwich-type) complex [Ag(2)(L)(3)](PF(6))(2) (4) on its reaction with AgPF(6). Each silver adopts a trigonal planar coordination environment with coordination sites occupied by two S atoms from one terminal ligand, and by one S atom from the bridging ligand; once again the ether oxygens are not coordinated. An NMR titration of the formation of complex 4 confirmed that the Ag/L stoichiometry of 2:3 found in the solid state is also maintained in solution. PMID- 19902907 TI - Palladium motion in cyclomeric compounds: a theoretical study. AB - Density functional theory (DFT)/B3LYP calculations have been carried out to study successive intramolecular 1,n palladium shifts (n = 3-5) in palladium complexes of organic cyclomers. Such shifts of the PdBr(phosphine) moiety, which is bound to the cyclomer and which exchanges concomitantly with a hydrogen atom distant by n carbon atoms from palladium, might lead to an endless motion around the cyclomer. The cyclomers that have been analyzed are either the [1,1,1,1]paracyclophane 1, a THF-based 16-crown-4 structure 2, or the THF ethylene cyclic dimer 3 (THF = tetrahydrofuran). We show that the [1,1,1,1]paracyclophane 1 is not a good candidate for a circular motion of the metallic moiety. This is due to the very high barrier of the 1,2 Pd/H exchange within the phenyl ring. Hence only a pendulum movement of the metallic moiety between two adjacent phenyl rings can easily take place. For the THF-based systems 2 and 3, the processes along the exo face are found to involve high energy barriers. Processes along the endo face are more accessible, especially for 2 where an endless motion made of successive 1,2 and 1,5 shifts is characterized by barriers that are somewhat less than 30 kcal mol(-1). PMID- 19902908 TI - Novel, high throughput method to study in vitro protein release from polymer nanospheres. AB - Controlled delivery of therapeutic protein drugs using biodegradable polymer carriers is a desired characteristic that enables effective, application-specific therapy and treatment. Previous studies have focused on protein delivery from polymers using conventional "one-sample-at-a-time" techniques, which are time consuming and costly. In addition, many therapeutic proteins are in limited supply and are expensive, so it is desirable to reduce sample size for design and development of delivery devices. We have developed a rapid, high throughput technique based on a highly sensitive fluorescence-based assay to detect and quantify protein released from polyanhydrides while utilizing relatively small amounts of protein (approximately 40 microg). These studies focused on the release of a model protein, Texas Red conjugated bovine serum albumin, from polyanhydride copolymers based on sebacic acid (SA) and 1,6-bis(p carboxyphenoxy)hexane (CPH). The protein release profiles were assessed simultaneously to investigate the effect of polymer device geometry (nanospheres vs films), polymer chemistry, and pH of the release medium. The results indicated that the nanosphere geometry, SA-rich chemistries, and neutral pH release medium led to a more rapid release of the protein compared to the film geometry, CPH rich chemistries, and acidic pH release medium, respectively. This high throughput fluorescence-based method can be readily extended to study release kinetics for other proteins and polymer systems. PMID- 19902909 TI - Proteins fold by subdiffusion of the order parameter. AB - It is shown that the folding of a C(alpha) model of chymotyprsin inhibitor (CI2) protein cannot be described by either diffusion (Smoluchowski equation, SE) or a normal-diffusion continuous time random walk of a single order parameter under the influence of the thermodynamic force. The reason for these failures is that the order parameter follows subdiffusion. A theory is proposed based on the idea that an ordinary SE holds along a contour representative of the folding pathways, and that displacements along the contour obey a fractal relationship to, and are longer than, those along the reaction coordinate defined by the order parameter. With a new, constraint-free method to determine the order-parameter-dependent diffusion constant, and statistical temperature molecular dynamics (STMD) enhanced sampling of the free energy, the fractal SE theory is completely characterized by short-time simulations, and its predictions are in quantitative agreement with simulated long-time folding dynamics. Thus, the fractal SE may serve as an accelerated algorithm to study the folding of proteins too slow to be simulated directly. PMID- 19902910 TI - Nature of excited states and relaxation mechanisms in C-phycocyanin. AB - The electronic structure and photoinduced relaxation dynamics of the cyanobacterial light harvesting protein, C-Phycocyanin (CPC), are examined using transient grating and two-dimensional (2D) photon echo spectroscopies possessing sub-20 fs time resolution. In combination with linear absorption and fluorescence measurements, these time-resolved experiments are used to constrain the parameters of a Frenkel exciton Hamiltonian. Particular emphasis is placed on elucidating the nature of excited states involving the alpha84 and beta84 phycocyanobilin pigment dimers of CPC. This paper obtains new experimental evidence suggesting that electronic relaxation proceeds by way of incoherent energy transfer between the alpha84 and beta84 pigment sites (i.e., the weak coupling limit of energy transfer). Transient absorption anisotropies simulated in the weak coupling limit agree well with measurements, whereas signals computed in an exciton basis possess short-lived (electronic) coherent components not present in the experimental data. In addition, 2D photon echo spectra for CPC show no sign of the interfering nonlinearities predicted by a theoretical model to be characteristic of exciton formation. Another important new observation is that the sub-100 fs dynamics in the transient absorption anisotropy are dominated by an impulsively excited hydrogen out-of-plane wagging mode similar to those observed in phytochrome and retinal. Detection of this 795 cm(-1) coherence is of particular interest because our recent study of a closely related protein, Allophycocyanin (APC), assigns a similar coordinate as a promoting mode enabling ultrafast internal conversion. Together, the experiments conducted for APC and CPC suggest that interactions between the pigments and environment are the key to understanding why electronic relaxation in CPC is more than three times slower than APC despite the nearly identical geometries of the pigment dimers. Most important in reaching this conclusion is the present finding that relaxation of the 2D photon echo line shapes of CPC is approximately two times faster than that measured for APC. Overall, the present results underscore the ability of phycobiliproteins to control light harvesting dynamics through solvation and variation in the conformations of open-chain tetrapyrrole chromophores. PMID- 19902911 TI - Surface "click" chemistry on brominated plasma polymer thin films. AB - A brominated plasma polymer (BrPP) thin film was fabricated on a variety of substrate surfaces (silicon wafers, glass, gold, and polymers) via the radio frequency glow discharge of 1-bromopropane. This BrPP thin film was highly adherent and stable and was found to be a useful platform for secondary reactions, leading to surfaces with specific chemical functionalities. Following nucleophilic exchange, an azide-functionalized PP thin film was prepared that was reactive toward two different alkynes via the copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction, a paradigm of "click" chemistry. "Click" microcontact printing (microCP) of a fluorescent alkyne was also successfully carried out, demonstrating the versatility and functionality of this new class of reactive thin film plasma polymer coatings. PMID- 19902912 TI - A theoretical study on the nature of on- and off-states of reversibly photoswitching fluorescent protein Dronpa: absorption, emission, protonation, and Raman. AB - A GFP-like fluorescent protein, Dronpa, which was engineered from a coral Pectiniidae, was found to display perfect photochromic properties; the fluorescent "on"-state and nonfluorescent "off"-state of Dronpa can be reversibly switched by irradiation of two different wavelengths of light. To understand the detailed mechanism of the reversible photoswitching process at the atomic level, we performed QM and ONIOM(QM:MM) calculations to study the nature of the proposed on-state and off-state. Several high-level QM methods (TD-B3LYP, CASSCF, CASPT2, and SAC-CI) were employed to compute the vertical absorption and emission energies in the gas phase for four different protonation states as well as two conformations. The vertical absorption and emission energies of the on- and off states in the proteins were further studied by the ONIOM(QM:MM) calculations. The ONIOM calculations on the absorption and emission suggest the neutral trans form is the off-state and the anionic cis form is the on-state. The dominant protonation states of the on- and off-states are also supported by protonation probability calculations via Poisson-Boltzmann electrostatics and Monte Carlo sampling. Moreover, the local protein environments were found to influence protonation states of the chromophore. Different possible reaction mechanisms are also discussed. PMID- 19902913 TI - New algorithm for the identification of intact disulfide linkages based on fragmentation characteristics in tandem mass spectra. AB - Identifying the sites of disulfide bonds in a protein is essential for thorough understanding of a protein's tertiary and quaternary structures and its biological functions. Disulfide linked peptides are usually identified indirectly by labeling free sulfhydryl groups with alkylating agents, followed by chemical reduction and mass spectral comparison or by detecting the expected masses of disulfide linked peptides on mass scan level. However, these approaches for determination of disulfide bonds become ambiguous when the protein is highly bridged and modified. For accurate identification of disulfide linked peptides, we present here an algorithmic solution for the analysis of tandem mass (MS/MS) spectra of disulfide bonded peptides under nonreducing condition. A new algorithm called "DBond" analyzes disulfide linked peptides based on specific features of disulfide bonds. To determine disulfide linked sites, DBond takes into account fragmentation patterns of disulfide linked peptides in nucleoside diphosphate kinase (NDPK) as a model protein, considering fragment ions including cysteine, cysteine thioaldehyde (-2 Da, C(T)), cysteine persulfide (+32 Da, C(S)) and dehydroalanine (-34 Da, C(Delta)). Using this algorithm, we successfully identified about a dozen novel disulfide bonds in a hexa EF-hand calcium binding protein secretagogin and in a methionine sulfoxide reductase. We believe that DBond, taking into account the disulfide bond fragmentation characteristics and post-translational modifications, offers a novel approach for automatic identification of unknown disulfide bonds and their sites in proteins from MS/MS spectra. PMID- 19902914 TI - Surface-enhanced transmission mode desorption electrospray ionization: increasing the specificity of ambient ionization mass spectrometric analyses. AB - Fabrication and utilization of mesh materials specifically designed to capture analytes from solution facilitates the direct coupling of affinity capture and ambient ionization mass spectrometry via surface-enhanced transmission mode desorption electrospray ionization (TM-DESI). Incorporation of photolabile groups within the linkage between the mesh surface and the covalently modified reactive probe affords facile release of mass tagged analytes directly to mesh surfaces that have been rinsed free of matrix interferences. The approach introduces increased specificity to the already rapid TM-DESI analysis technique, resulting in a powerful tool for high-throughput screening of targeted analytes. Specific capture of thiols is discussed herein, but the surface-enhanced TM-DESI technique can be readily extended to other functional groups by alteration of the capture agent. PMID- 19902915 TI - Heavy alkali metal amides: role of secondary interactions in metal stabilization. AB - The coordination chemistry of the bis(diphenylmethylsilyl)amine ligand, HN(SiMePh(2))(2), with the heavy alkali metals potassium and rubidium has been investigated to study its effect on the structure of the resulting compounds. The compounds exhibit extensive intra- and intermolecular M-pi interactions, creating 1-D coordination polymers, [K{N(SiMePh(2))(2)}](infinity) (1) and [Rb{N(SiMePh(2))(2)}](infinity) (3). This motif is maintained in the presence of tetrahydrofuran (THF), as seen in [K{N(SiMePh(2))(2)}thf](infinity) (2). In contrast, use of the tridentate PMDTA allows isolation of a monomeric species, K(pmdta)N(SiMePh(2))(2) (4), which exhibits both agostic and M-pi interactions. The incorporation of macrocyclic 18-crown-6, results in the formation of a rare non-metal bound amido species, {[K(18-crown-6)N(SiMePh(2))(2)]}(infinity) (5), stabilized by M-pi interactions from adjacent [N(SiMePh(2))(2)](-) ligands propagating 1-D coordination polymers. PMID- 19902916 TI - Synthesis and optical properties of guanosine 5'-monophosphate-mediated CdS nanostructures: an analysis of their structure, morphology, and electronic properties. AB - The present manuscript reports the synthesis, characterization, and analysis of electronic properties of water-soluble guanosine 5'-monophosphate (GMP)-mediated CdS quantum dots (Q-dots). The morphology, size, and size distribution of these particles have been analyzed by transmission electron microscopy. These particles display the onset of absorption at 2.7 eV and emission at 2.2 eV. In comparison to other monophosphates of RNA (AMP, UMP, and CMP), GMP-mediated CdS exhibits enhanced electronic properties. The participation of different functional groups of GMP in the stabilization of CdS nanoparticles has been analyzed by FTIR and (1)H and (31)P NMR spectroscopic techniques. Two types of binding sites involving phosphorus centers are indicated by IR and (31)P NMR studies. The conversion of CdS Q-dots to nanorods has been monitored by using electron microscopy, steady state optical and fluorescence measurements, and a fluorescence lifetime system coupled with anisotropy accessories. The observed change in the morphology and electronic behavior of GMP- and RNA-mediated CdS nanostructures is discussed on the basis of their structural difference. PMID- 19902917 TI - Endosomal leakage and nuclear translocation of multiwalled carbon nanotubes: developing a model for cell uptake. AB - We report our findings on cellular membrane penetration, endocytosis, endosomal leakage, and nuclear translocation of multiwalled carbon nanotubes (MWCNTs). Our data is consistent with a working model for MWCNTs' cell uptake and cellular translocations. PMID- 19902918 TI - Direct correlation of structural domain formation with the metal insulator transition in a VO2 nanobeam. AB - The electrical resistance of single VO(2) nanobeams was measured while simultaneously mapping the domain structure with Raman spectroscopy to investigate the relationship between structural domain formation and the metal insulator transition. With increasing temperature, the nanobeams transformed from the insulating monoclinic M(1) phase to a mixture of the Mott-insulating M(2) and metallic rutile phases. Domain fractions were used to extract the temperature dependent resistivity of the M(2) phase, which showed an activated behavior consistent with the expected Mott-Hubbard gap. Metallic monoclinic phases were also produced by direct injection of charge into devices, decoupling the Mott metal-insulator transition from the monoclinic to rutile structural phase transition. PMID- 19902919 TI - Detection of local protein structures along DNA using solid-state nanopores. AB - Nanopores have been successfully employed as a new tool to rapidly detect single biopolymers, in particular DNA. When a molecule is driven through a nanopore by an externally applied electric field, it causes a characteristic temporary change in the trans-pore current. Here, we examine the translocation of DNA with discrete patches of the DNA-repair protein RecA attached along its length. Using the fact that RecA-coated DNA and bare DNA yield very different current-blockade signatures, we demonstrate that it is possible to map the locations of the proteins along the length of a single molecule using a solid-state nanopore. This is achieved at high speed and without any staining. We currently obtain a spatial resolution of about 8 nm, or 5 RecA proteins binding to 15 base pairs of DNA, and we discuss possible extensions to single protein resolution. The results are a crucial first step toward genomic screening, as they demonstrate the feasibility of reading off information along DNA at high resolution with a solid-state nanopore. PMID- 19902920 TI - Using the antenna effect as a spectroscopic tool: photophysics and solution thermodynamics of the model luminescent hydroxypyridonate complex [Eu(III)(3,4,3 LI(1,2-HOPO))]-. AB - Although widely used in bioassays, the spectrofluorimetric method described here uses the antenna effect as a tool to probe the thermodynamic parameters of ligands that sensitize lanthanide luminescence. The Eu(3+) coordination chemistry, solution thermodynamic stability, and photophysical properties of the spermine-based hydroxypyridonate octadentate chelator 3,4,3-LI(1,2-HOPO) are reported. The complex [Eu(III)(3,4,3-LI(1,2-HOPO))](-) luminesces with a long lifetime (805 mus) and a quantum yield of 7.0% in aqueous solution, at pH 7.4. These remarkable optical properties were exploited to determine the high (and proton-independent) stability of the complex (log beta(110) = 20.2(2)) and to define the influence of the ligand scaffold on the stability and photophysical properties. PMID- 19902921 TI - Solar fuels via artificial photosynthesis. AB - Because sunlight is diffuse and intermittent, substantial use of solar energy to meet humanity's needs will probably require energy storage in dense, transportable media via chemical bonds. Practical, cost effective technologies for conversion of sunlight directly into useful fuels do not currently exist, and will require new basic science. Photosynthesis provides a blueprint for solar energy storage in fuels. Indeed, all of the fossil-fuel-based energy consumed today derives from sunlight harvested by photosynthetic organisms. Artificial photosynthesis research applies the fundamental scientific principles of the natural process to the design of solar energy conversion systems. These constructs use different materials, and researchers tune them to produce energy efficiently and in forms useful to humans. Fuel production via natural or artificial photosynthesis requires three main components. First, antenna/reaction center complexes absorb sunlight and convert the excitation energy to electrochemical energy (redox equivalents). Then, a water oxidation complex uses this redox potential to catalyze conversion of water to hydrogen ions, electrons stored as reducing equivalents, and oxygen. A second catalytic system uses the reducing equivalents to make fuels such as carbohydrates, lipids, or hydrogen gas. In this Account, we review a few general approaches to artificial photosynthetic fuel production that may be useful for eventually overcoming the energy problem. A variety of research groups have prepared artificial reaction center molecules. These systems contain a chromophore, such as a porphyrin, covalently linked to one or more electron acceptors, such as fullerenes or quinones, and secondary electron donors. Following the excitation of the chromophore, photoinduced electron transfer generates a primary charge-separated state. Electron transfer chains spatially separate the redox equivalents and reduce electronic coupling, slowing recombination of the charge-separated state to the point that catalysts can use the stored energy for fuel production. Antenna systems, employing a variety of chromophores that absorb light throughout the visible spectrum, have been coupled to artificial reaction centers and have incorporated control and photoprotective processes borrowed from photosynthesis. Thus far, researchers have not discovered practical solar-driven catalysts for water oxidation and fuel production that are robust and use earth-abundant elements, but they have developed artificial systems that use sunlight to produce fuel in the laboratory. For example, artificial reaction centers, where electrons are injected from a dye molecule into the conduction band of nanoparticulate titanium dioxide on a transparent electrode, coupled to catalysts, such as platinum or hydrogenase enzymes, can produce hydrogen gas. Oxidizing equivalents from such reaction centers can be coupled to iridium oxide nanoparticles, which can oxidize water. This system uses sunlight to split water to oxygen and hydrogen fuel, but efficiencies are low and an external electrical potential is required. Although attempts at artificial photosynthesis fall short of the efficiencies necessary for practical application, they illustrate that solar fuel production inspired by natural photosynthesis is achievable in the laboratory. More research will be needed to identify the most promising artificial photosynthetic systems and realize their potential. PMID- 19902922 TI - Electrochemical and photovoltaic properties of electropolymerized poly(thienylsilole)s. AB - Electrochemical and photoelectrochemical properties were studied of a series of donor-acceptor materials based on polythiophene modified with silole moieties. The materials were prepared by electrochemical anodic polymerization of 2,5 bis([2,2'-bithiophen]-5-yl)-1,1-dimethyl-3,4-diphenylsilole and 2,5-bis([2,2' terthiophen]-5-yl)-1,1-dimethyl-3,4-diphenylsilole, as well as copolymerization of these monomers with 2,2'-bithiophene. Photocurrent measurements showed that introduction of silole resulted in a considerable enhancement of the photovoltaic properties of silole-containing materials and especially the fill factor. However, as demonstrated by Mott-Schottky measurements, electropolymerized silole containing materials showed a substantial degree of disorder and high density of states in the midgap, which negatively affected their photovoltaic properties. Atomic force microscopy (AFM) and phase imaging revealed the presence of phase segregation and heterogeneity of the silole-containing materials. Interestingly, introduction of siloles suppressed the cathodic (n-type) doping typical for polythiophenes. This work demonstrates that siloles show great promise as electron-acceptor groups for all-organic solar cells; however, further work is required to optimize the properties and performance of poly(thienylsilole)-based materials. PMID- 19902923 TI - Synchronously amplified fluorescence image recovery (SAFIRe). AB - Fluorescence intermittency severely limits brightness in both single molecule and bulk fluorescence. Herein, we demonstrate that optical depopulation of organic fluorophore triplet states opens a path to significantly increased sensitivity by simultaneously increasing brightness and greatly reducing background through synchronously detected fluorescence modulation. Image recovery is achieved through selective fluorescence enhancement via modulating a secondary laser excitation at much lower energy than the observed emission in order to depopulate the long-lived triplet states. A series of xanthene dyes that exhibit efficient triplet-state formation demonstrate that this method of selective signal extraction can be achieved at moderate primary and secondary excitation intensities through tailoring dye photophysics and imaging conditions. Up to 5 fold increases in solution-based fluorescence over primary laser excitation alone was achieved upon secondary laser excitation, and dynamic control of signal modulation was demonstrated over a wide time range simply by varying the modulation frequency of the laser used for depopulation of the triplet state. We identify the photophysical characteristics that enable existing or to-be-designed fluorophores to be used in synchronously amplified fluorescence image recovery (SAFIRe) microscopy. PMID- 19902924 TI - Spontaneous deformation of an oil droplet induced by the cooperative transport of cationic and anionic surfactants through the interface. AB - Spontaneous deformation of a tetradecane droplet with palmitic acid on an aqueous phase with stearyltrimethylammonium chloride is reported. Palmitic acid is transported from the oil droplet to the aqueous phase by the concentration difference between the organic and the aqueous phases. The transport of palmitic acid causes the oil droplet interface to undergo various spontaneous deformations. When the oil droplet is placed on an aqueous surface, its diameter shrinks. Several tens of seconds later, the oil droplet suddenly expands and then shrinks in a second. After such a dramatic deformation, the oil droplet undergoes blebbing on its oil-water interface for over 1 h. We investigated the physicochemical mechanism of these phenomena. We discuss the cause of these deformations in terms of the spatiotemporal variation of the interfacial tension and elucidate that the blebbing deformation is due to the surfactant aggregate generated by cationic and anionic surfactants. PMID- 19902925 TI - Capillary electrophoresis with electrochemiluminescent detection for highly sensitive assay of genetically modified organisms. AB - A capillary electrophoresis coupled with electrochemiluminescent detection system (CE-ECL) was developed for the detection of polymerase chain reaction (PCR) amplicons. The ECL luminophore, tris(1,10-phenanthroline) ruthenium(II) (Ru(phen)(3)(2+)), was labeled to the PCR primers before amplification. Ru(phen)(3)(2+) was then introduced to PCR amplicons by PCR amplification. Eventually, the PCR amplicons were separated and detected by the homemade CE-ECL system. The detection of a typical genetically modified organism (GMO), Roundup Ready Soy (RRS), was shown as an example to demonstrate the reliability of the proposed approach. Four pairs of primers were amplified by multiple PCR (MPCR) simultaneously, three of which were targeted on the specific sequence of exogenous genes of RRS, and another was targeted on the endogenous reference gene of soybean. Both the conditions for PCR amplification and CE-ECL separation and detection were investigated in detail. Results showed that, under the optimal conditions, the proposed method can accurately identifying RRS. The corresponding limit of detection (LOD) was below 0.01% with 35 PCR cycles. PMID- 19902926 TI - Detection of protein binding using activator generated by electron transfer for atom transfer radical polymerization. AB - A purge-free controlled living polymerization method, activator generated by electron transfer for atom transfer radical polymerization (AGET ATRP), is used to amplify the occurrence of protein binding events. Detection of ovalbumin is demonstrated where binding of femtomole protein is differentiable from the background using ellipsometry. Moreover, binding of subpicomole protein leads to visually distinguishable spots on the sensor surface within 15 min, which signifies its potential in future development of point-of-need devices. PMID- 19902927 TI - Photochemical reactivity of graphene. AB - We demonstrate a photochemical reaction between graphene and benzoyl peroxide. This reaction introduces spatially localized defects into the graphene basal plane. The reactivity of the single-layer graphene is approximately 14 times higher than that of the double-layer graphene. Our result suggests that photoexcited graphene transfers a hot electron to benzoyl peroxide and induces its decomposition to a phenyl radical. PMID- 19902928 TI - Biological implications of polymeric microdevices for live cell assays. AB - Lab-on-a-chip technologies have the potential to deliver significant technological advances in modern biomedicine, through the ability to provide appropriate low-cost microenvironments for screening cells. However, to date, few studies have investigated the suitability of poly(dimethylsiloxane) (PDMS) for live cell culture. Here, we describe an inexpensive method for production of reusable, optical-grade PDMS microculture chips which provide a static and self contained microwell system analogous to conventional polystyrene multiwell plates. We use these structures to probe the effects of PDMS upon live cell culture bioassays, using time-lapse fluorescence imaging to explore the toxicity of the substrate. We use three model systems to explore the efficacy of the microstructured devices: (i) live cell culture, (ii) adenoviral gene delivery to mammalian cells, and (iii) gravity enforced formation of multicellular tumor spheroids (MCTS). Results show that PDMS is nontoxic to cells, as their viability and growth characteristic in PDMS-based platforms is comparable to that of their polystyrene counterparts. PMID- 19902929 TI - Carbohydrate-protein interactions investigated on plastic chips statically coated with hydrophobically modified hydroxyethylcellulose. AB - We developed a novel method for rapid screening of carbohydrate-protein interactions using poly(methyl methacrylate) (PMMA) channels statically coated with hydrophobically modified hydroxyethylcellulose (HM-HEC). We found that a self-assembled monolayer (SAM) of HM-HEC on a PMMA surface intact by water allows rapid and reproducible separations of glycan samples using a 20 mM phosphate without HM-HEC. The underlying mechanism for dynamic and static coatings on the PMMA surface is discussed. Simultaneous analysis of the molecular interaction between a complex mixture of carbohydrates from alpha1-acid glycoprotein and proteins has been successfully achieved in PMMA channels statically coated with a SAM of HM-HEC. PMID- 19902930 TI - Enzyme-induced staining of biomembranes with voltage-sensitive fluorescent dyes. AB - We consider the physicochemical basis for enzyme-induced staining of cell membranes by fluorescent voltage-sensitive dyes, a method that may lead to selective labeling of genetically encoded nerve cells in brain for studies of neuronal signal processing. The approach relies on the induction of membrane binding by enzymatic conversion of a water-soluble precursor dye. We synthesized an amphiphilic hemicyanine dye with and without an additional phosphate appendix at its polar headgroup. The fluorescence of these dyes is negligible in water but high when bound to lipid membranes. By fluorescence titration with lipid vesicles it was shown that the phosphate group lowers the partition coefficient from water to membrane by more than an order of magnitude. By isothermal titration calorimetry, we showed that the dye phosphate was a substrate for a water-soluble alkaline phosphatase following MichaelisMenten kinetics. In a suspension of lipid vesicles, the enzyme reaction led to a fluorescence increase due to enhanced membrane binding of the product dye in accord with the MichaelisMenten kinetics of the reaction and the partition coefficients of substrate and product. We successfully tested the staining method by fluorescence microscopy with individual giant lipid vesicles and with individual red blood cells. In both systems, the membrane fluorescence due to bound hemicyanine was enhanced by an order of magnitude, proving the feasibility of enzyme-induced staining with voltage-sensitive dyes. PMID- 19902931 TI - Stoichiometric quantification of Akt phosphorylation using LC-MS/MS. AB - The Ptdlns-3-kinase (PI3-K) signaling pathway plays a vital role in cell survival, proliferation, apoptosis and differentiation in normal cells, as well as in diseases such as cancer and diabetes. Quantification of phospho-Akt is a standard way of assessing the activity of the PI3-K signaling pathway in both cells and tumors. This measurement is traditionally performed semiquantitatively using immunoassays such as Western blot. Here we report an LC-MS method to accurately measure the stoichiometry of Akt phosphorylation in biological samples. The procedure includes immunoprecipitation, gel electrophoresis, in-gel digestion, addition of isotopicaly labeled internal standards and LC-MS/MS. Two proteolytic enzymes, chymotrypsin and trypsin, were used to generate suitable peptide fragments for measuring Thr308 and Ser473 phosphorylation, respectively. The interday imprecision was estimated to be 3.8% and 2.3% for Thr308 and Ser473, respectively. This method has been tested on human T-cells grown in presence and absence of pervanadate and with or without a PI3-K inhibitor and on human glioblastoma cells (U-87 MG) grown in presence and absence of wortmannin (PI3-K inhibitor).The results of T cells suggest that the levels of Akt phosphorylation in untreated cells were below 1% for both phosphorylation sites. Pervanadate treatment provoked an 18-fold increase in phosphorylation of Thr308 and the PI3-K inhibitor partially reversed the increase. A comparison between LC-MS/MS and Western blotting suggests that the LC-MS based method is of comparable sensitivity and provides a more accurate phosphorylation stoichiometry, a wider dynamic range and more in-depth information. The application of the new method and its utility to providing predictive markers of response to targeted therapies is discussed. PMID- 19902932 TI - Microfluidic analysis of complex samples with minimal sample preparation using gradient elution moving boundary electrophoresis. AB - Sample-in answer-out analytical tools remain the goal of much lab on a chip research, but miniaturized methods capable of examining minimally prepared samples have proven elusive. Complex samples, including whole milk, various types of dirt and leaves, coal fly ash, and blood serum, were analyzed quantitatively for dissolved potassium, calcium, sodium, magnesium, lithium, and melamine using gradient elution moving boundary electrophoresis (GEMBE) and contactless conductivity detection with the single preparatory step of dilution or suspension in sample buffer. GEMBE is a simple, robust analytical technique, well-suited to microfluidic analysis of complex samples containing material, such as particulates or proteins, that would confound the majority of other microfluidic techniques. GEMBE utilizes electrophoretic flow to drive electrically charged analytes into a microfluidic channel or capillary for detection, while opposing electro-osmotic and variable pressure-driven flows prevent the remainder of the sample from entering the channel. Contactless conductivity detection further simplifies device construction and operation, positioning GEMBE for inexpensive and facile multiplexed implementation outside laboratory settings. PMID- 19902933 TI - Unusual bonding in trisilaallene and related heavy allenes. AB - The synthesis, X-ray and spectroscopic analysis, and unusual bonding and structure among heavy group-14 element congeners of allene (heavy allenes) [R(2)M=M'=MR(2); M = M' = Si (1a), M = Si, M' = Ge (1b), M = Ge, M' = Si (1c), and M = M' = Ge (1d)] are comparatively discussed. Using DFT calculations and PMO theory, the origin of the bent and fluxional skeleton of the heavy allenes is ascribed to the Jahn-Teller distortion associated with the effective pi-sigma* mixing (pi-sigma* distortion). This type of distortion is suggested to be a key concept for qualitative description of the unusual bonding of unsaturated compounds of heavy main-group elements. PMID- 19902934 TI - Enzyme-based NAND and NOR logic gates with modular design. AB - The logic gates NAND/NOR were mimicked by enzyme biocatalyzed reactions activated by sucrose, maltose and phosphate. The subunits performing AND/OR Boolean logic operations were designed using maltose phosphorylase and cooperative work of invertase/amyloglucosidase, respectively. Glucose produced as the output signal from the AND/OR subunits was applied as the input signal for the INVERTER gate composed of alcohol dehydrogenase, glucose oxidase, microperoxidase-11, ethanol and NAD(+), which generated the final output in the form of NADH inverting the logic signal from 0 to 1 or from 1 to 0. The final output signal was amplified by a self-promoting biocatalytic system. In order to fulfill the Boolean properties of associativity and commutativity in logic networks, the final NADH output signal was converted to the initial signals of maltose and phosphate, thus allowing assembling of the same standard units in concatenated sequences. The designed modular approach, signal amplification and conversion processes open the way toward complex logic networks composed of standard elements resembling electronic integrated circuitries. PMID- 19902935 TI - Electrochemical release of fluorescently labeled thiols from patterned gold surfaces. AB - Reductive desorption of alkanethiols is a tool for spatially and temporally controlled release of small molecules or particles from individually addressable gold electrodes. Here we report on the dynamics of release using fluorophore terminated C6 or C11 thiols. We show that the release kinetics for C6 thiols are determined solely by diffusive transport, whereas for C11 thiols the release kinetics are attenuated by the low solubility that limits the rate at which the desorbed thiols can diffuse away from the surface. The release of multiple different molecules from the same electrode is demonstrated using red- and green emitting fluorophores. The fraction of the monolayer released is dependent on the electrode potential. PMID- 19902936 TI - Flexible and compressible Goretex-PEDOT membrane electrodes for solid-state dye sensitized solar cells. AB - A porous, flexible electrode based on a PTFE (Teflon) membrane (Goretex) coated with a metallic current collector and a conducting polymer (poly(3,4 ethylenedioxythiophene), PEDOT) has been developed for applications in solid state dye-sensitized solar cells. Its low sheet resistance and compressibility make it an ideal electrode on uneven TiO(2) surfaces with high efficiency and reproducibility. The porous nature of the electrode enables the feed-through of reactants and treatment agents, which opens up exciting opportunities to interface these photoelectrochemical devices with electrocatalytic, energy conversion, and storage systems. Postfabrication bonding of the photoanode and the Goretex-Au-PEDOT electrode is demonstrated. PMID- 19902937 TI - Structure-activity relationships for the degradation reaction of 1-beta-O-acyl glucuronides. Part 3: Electronic and steric descriptors predicting the reactivity of aralkyl carboxylic acid 1-beta-O-acyl glucuronides. AB - Since 1-beta-O-acyl glucuronides (betaGAs) are thought to be chemically reactive metabolites capable of binding to tissue proteins, possibly leading to adverse drug reactions of the parent carboxylic acid drugs, we have initiated research efforts to derive structure-activity relationships (SARs) of betaGAs, with a focus on finding appropriate descriptors that predict their intrinsic electrophilic reactivity or degradation rate constants (k values). Our previous SAR studies on the k values of betaGAs derived from o-, m-, and p-substituted benzoic acids demonstrated that the diversity and complexity of the k values were controlled by the electronic and/or steric effects of the parent carboxylic acids. In the present study, we performed further SAR studies on the k values of 13 betaGAs derived from aralkyl carboxylic acids, focusing on the substituents and stereochemistry at the alpha-position of the parent carboxylic acids. In single regression analyses, the pKa and (1)H NMR chemical shifts (delta(COOH)) of the parent carboxylic acids correlated well with the log k values of seven betaGAs derived from five arylacetic and two (R)-2-arylpropionic acids, whereas the (13)C NMR chemical shifts [delta(C horizontal lineO) and delta((C horizontal lineO)OH)] correlated with the log k values of another seven betaGAs derived from the five arylacetic and two (S)-2-arylpropionic acids. Excellent correlations were also obtained between the log k values of four betaGAs with a common 4 phenylbenzyl moiety and the partial atomic charges (natural type) of the corresponding carboxylic hydrogen atoms (Hpac), the molar volume (MV), and the molar refractivity (MR). In multiple regression analyses, appropriate combinations of electronic (delta(COOH) or pKa) and steric [Taft's steric constant (Es) or delta((C horizontal lineO)OH)] descriptors could predict the log k values of betaGAs; electron-withdrawing 1-beta-O-acyl groups increased the k values, while increasing steric hindrance around the linkages decreased them. The standard partial regression coefficients indicated that the steric effects of the 1-beta-O-acyl groups of betaGAs affected the k values as strongly as the electronic effects. External validation of the derived SAR models is also discussed. PMID- 19902938 TI - Theoretical investigation of mechanisms for the gas-phase unimolecular decomposition of DMMP. AB - All species involved in the multichannel decomposition of gas-phase dimethyl methylphosphonate (DMMP) were investigated by electronic structure calculations. Geometries for stationary structures along the reaction paths, were fully optimized with the MP2 method and the B3LYP and MPW1K DFT functionals, and the 6 31G*, 6-31++G**, and aug-cc-pVDZ basis sets. The geometries determined by the B3LYP and MPW1K functionals are in very good agreement with the MP2 values. Increasing the basis set size from 6-31G* to aug-cc-pVDZ does not significantly alter this result. Single point energy calculations were carried out with highly accurate but computationally more expensive CBS-QB3 theory. DMMP has three conformers, which lead to the four primary product channels, (O)P(CH(2))(OCH(3)) + CH(3)OH, (O)P(CH(3)) (OCH(3))(OH) + CH(2), c-(O)P(CH(3))OCH(2) + CH(3)OH, and (O)P(CH(3))(OCH(3))(OCH) + H(2). The first channel has the lowest energy barrier and is expected to be the most important pathway. It occurs via C-H and P-O bond cleavages accompanied by O-H bond formation. The other three channels have higher and similar energy barriers, and are expected to have smaller and similar rates. The product (O)P(CH(3))(OCH(3))(OCH) undergoes a secondary decomposition to form (OH)P(CH(3))(OCH(3)) + CO. PMID- 19902939 TI - A stereoselective approach to nucleosides and 4'-thioanalogues from acyclic precursors. AB - D- and L-nucleosides and analogues thereof, including the 4'-thionucleoside series, are one of the most important biological and pharmaceutically active classes of compounds. A novel approach to their synthesis from chiral acyclic thioaminal, bearing the nucleobase, is described. PMID- 19902940 TI - Alternating pattern of stereochemistry in the nonactin macrocycle is required for antibacterial activity and efficient ion binding. AB - Nonactin is a polyketide antibiotic produced by Streptomyces griseus ETH A7796 and is an ionophore that is selective for K(+) ions. It is a cyclic tetraester generated from two monomers of (+)-nonactic acid and two of (-)-nonactic acid, arranged (+)-(-)-(+)-(-) so that nonactin has S4 symmetry and is achiral. To understand why achiral nonactin is the naturally generated diastereoisomer, we generated two alternate diastereoisomers of nonactin, one prepared solely from (+)-nonactic acid and one prepared solely from (-)-nonactic acid, referred to here as 'all-(+)-nonactin' and 'all-(-)-nonactin', respectively. Both non-natural diastereoisomers were 500-fold less active against gram positive organisms than nonactin confirming that the natural stereochemistry is necessary for biological activity. We used isothermal calorimetry to obtain the K(a), DeltaG, DeltaH, and DeltaS of formation for the K(+), Na(+), and NH(4)(+) complexes of nonactin and all-(-)-nonactin; the natural diastereoisomer bound K(+) 880-fold better than all (-)-nonactin. A picrate partitioning assay confirmed that all-(-)-nonactin, unlike nonactin, could not partition K(+) ions into organic solvent. To complement the thermodynamic data we used a simple model system to show that K(+) transport was facilitated by nonactin but not by all-(-)-nonactin. Modeling of the K(+) complexes of nonactin and all-(-)-nonactin suggested that poor steric interactions in the latter complex precluded tight binding to K(+). Overall, the data show that both enantiomers of nonactic acid are needed for the formation of a nonactin diastereoisomer that can act as an ionophore and has antibacterial activity. PMID- 19902941 TI - Synthesis of imogolite fibers from decimolar concentration at low temperature and ambient pressure: a promising route for inexpensive nanotubes. AB - To date, the successful low-temperature synthesis of the aluminosilicate imogolite nanotubes always involved initial concentrations of the reagents in the millimolar range, higher concentrations being reported to lead to the formation of the less well characterized allophane phase. The present work shows that reaction kinetics and not initial concentration control the formation of the nanotubes: substantial amounts of well formed imogolite were obtained from a decimolar initial concentration, i.e. 100 times higher than the "standard" protocol. The allophane-like spheroids expected from the high reagent concentration were not observed in this work, and proto-imogolite was obtained instead. PMID- 19902943 TI - New factor characterizing the in-mouth release of odorants (volatile thiols): compositional changes in odorants exhaled from the human nose during drinking. AB - The aim of this research was to investigate the relationship between the odorants in the model drink and the odorants reaching the olfactory epithelium using the Retronasal Flavor Impression Screening System (R-FISS). By application of the R FISS to the odorants in the model drink, it was found that a methylthio ether [1 methoxy-3-methyl-3-(methylthio)butane] was detected with the original volatile thiol (4-methoxy-2-methyl-2-mercaptobutane) in the air exhaled through the nostrils via the nasal cavity after the model drink (including the original thiol) had been swallowed. In addition, this phenomenon was also observed in other volatile thiols (furfuryl mercaptan, ethyl 2-mercaptopropionate, 2-methyl-1 mercaptobutane, and 4-mercapto-4-methyl-2-pentanone). These compositional changes in thiols that were observed in the air exhaled through the nostrils could be affected by the chemical structure of each tested thiol and individual differences. These results pointed to a possibility that the odorants reaching the olfactory receptor via the throat during consumption of foods could not always retain their original chemical structures and compositions in foodstuffs. Therefore, the characteristic odor of volatile thiols might be perceived due to the stimuli of multiple compounds. To understand in detail flavor perception during the consumption of foods, not only the compositions or amounts of odorants in foodstuffs but also the compositional changes in odorants induced by biological reactions (reduction or methylation) need to be taken into consideration. PMID- 19902942 TI - Palladium-catalyzed enantioselective addition of two distinct nucleophiles across alkenes capable of quinone methide formation. AB - A sequential intramolecular-intermolecular enantioselective alkene difunctionalization reaction has been developed which is thought to proceed through Pd-catalyzed quinone methide formation. The synthesis of new chiral heterocyclic compounds with adjacent chiral centers is achieved in enantiomeric ratios up to 99:1 and diastereomeric ratios up to 10:1. PMID- 19902944 TI - Simultaneous removal of oxytetracycline and sulfamethazine antibacterials from animal waste by chemical oxidation processes. AB - Simultaneous degradation of oxytetracycline (OTC) and sulfamethazine (SMZ) antibacterials in synthetically contaminated cow manure (20 mg of antibacterials/kg of manure) in the presence and absence of bedding was investigated by the application of ozone, Fenton, and persulfate oxidation processes. Almost the complete removal of antibacterials was attained with all oxidation processes, which were combined with a pretreatment of manure with magnesium (Mg(2+)) salt desorption. Among the investigated oxidation processes, thermally activated persulfate oxidation with 25 mM Na(2)S(2)O(8) at 50 degrees C was also applied to the animal feeding operations wastewater, and the pseudo first-order degradation rate constants of OTC and SMZ were found as 3.22 and 1.25 (1/h), respectively. Thermally activated persulfate treatment resulted in the reduction of 82% inhibition of OTC and SMZ to 7%, indicating the production of almost nontoxic degradation products in the wastewater. PMID- 19902945 TI - Gas-phase reactivity of protonated 2-, 3-, and 4-dehydropyridine radicals toward organic reagents. AB - To explore the effects of the electronic nature of charged phenyl radicals on their reactivity, reactions of the three distonic isomers of n-dehydropyridinium cation (n = 2, 3, or 4) have been investigated in the gas phase by using Fourier transform ion cyclotron resonance mass spectrometry. All three isomers react with cyclohexane, methanol, ethanol, and 1-pentanol exclusively via hydrogen atom abstraction and with allyl iodide mainly via iodine atom abstraction, with a reaction efficiency ordering of 2 > 3 > 4. The observed reactivity ordering correlates well with the calculated vertical electron affinities of the charged radicals (i.e., the higher the vertical electron affinity, the faster the reaction). Charged radicals 2 and 3 also react with tetrahydrofuran exclusively via hydrogen atom abstraction, but the reaction of 4 with tetrahydrofuran yields products arising from nonradical reactivity. The unusual reactivity of 4 is likely to result from the contribution of an ionized carbene-type resonance structure that facilitates nucleophilic addition to the most electrophilic carbon atom (C-4) in this charged radical. The influence of such a resonance structure on the reactivity of 2 is not obvious, and this may be due to stabilizing hydrogen-bonding interactions in the transition states for this molecule. Charged radicals 2 and 3 abstract a hydrogen atom from the substituent in both phenol and toluene, but 4 abstracts a hydrogen atom from the phenyl ring, a reaction that is unprecedented for phenyl radicals. Charged radical 4 reacts with tert-butyl isocyanide mainly by hydrogen cyanide (HCN) abstraction, whereas CN abstraction is the principal reaction for 2 and 3. The different reactivity observed for 4 (as compared to 2 and 3) is likely to result from different charge and spin distributions of the reaction intermediates for these charged radicals. PMID- 19902946 TI - Self-assembly and structure of directly imaged inorganic-anion monolayers on a gold nanoparticle. AB - Cryogenic "trapping" was used to obtain the first TEM images of self-assembled monolayers of inorganic anions on a gold nanoparticle. This unique structural information makes it possible to study the formation of a protecting-ligand shell at an unprecedented level of detail. The protecting ligands are polyoxometalates (POMs; alpha-X(n+)W(12)O(40)((8-n)-), X(n+) = Al(3+) and "2H(+)", and alpha X(n+)W(11)O(39)((12-n)-), X(n+) = P(5+), Si(4+), and Al(3+)) with large negative charges for association with the gold surface and W atoms (Z = 74) for TEM imaging. The POM-anion shells were obtained by ligand exchange from citrate protected 13.8 nm gold nanoparticles. Replacement of the organic (citrate) by inorganic (tungsten-oxide) ligand shells results in substantial changes in the surface plasmon resonance (SPR). By correlating cryo-TEM images with changes in the SPR, degrees of surface coverage were reliably quantified by UV-visible spectroscopy. Then, the kinetics and thermodynamics of ligand-shell formation were investigated by systematically varying POM structure and charge. Rates of POM association with the gold surface ("nucleation") are inhibited by the electric-potential barrier of the citrate-stabilized particles, while binding affinities increase linearly with the charges (from 5- to 9-) of structurally different POM anions, suggesting that no single orientation ("lattice matching") is required for monolayer self-assembly. Time-dependent cryo-TEM images reveal that monolayer growth occurs via "islands", a mechanism that points to cation mediated attraction between bound POMs. Complete ligand shells comprised of 330 molecules of alpha-AlW(11)O(39)(9-) (1) possess small net charges (29e from zeta potential measurements) and short Debye lengths (kappa(-1) = 1.0 nm), which indicate that approximately 99% of the 2970 K(+) counter cations lie within ca. 1.5 nm (approximately 3 hydrated K(+) ion diameters) from the outer surface of the POM shell. Energetic analysis of the 1.57 +/- 0.04 nm center-to-center distance between molecules of 1 further indicates that K(+) ions reside in the ca. 4.5 A spaces between the bound ligands. These findings reveal an important structural role for counter cations within POM ligand shells on gold nanoparticles, analogous to that for cations in the monolayer walls of hollow POM macroanion vesicles. PMID- 19902947 TI - Evaluation of the effect of dietary virgin olive oil on blood pressure and lipid composition of serum and low-density lipoprotein in elderly type 2 diabetic subjects. AB - Dietary virgin olive oil may help to reduce blood pressure in hypertensive individuals, but little is known about the effect on type 2 diabetic patients. For the present study, 17 type 2 diabetic elderly subjects and 23 healthy elderly controls received a diet rich in virgin olive oil for 4 weeks. Blood pressure, biochemical parameters, low-density lipoprotein (LDL), and oxidized LDL lipids and fatty acids were measured. Systolic blood pressure was reduced after virgin olive oil consumption in both controls and diabetic patients. Although the biochemical parameters were not modified, the intervention protected LDL from oxidation and restored the levels of dihomo-gamma-linolenic acid (20:3, n-6) in serum cholesterol esters and phospholipids of diabetic patients. In conclusion, the present study provides new evidence of the effects of dietary virgin olive oil on blood pressure and LDL oxidation in type 2 diabetics. It is likely that the components responsible for the observed effects are the monounsaturated fatty acids and the presence of antioxidants in the oil, but this needs further investigation. PMID- 19902948 TI - Repellency of essential oils of Cryptomeria japonica (Pinaceae) against adults of the mosquitoes Aedes aegypti and Aedes albopictus (Diptera:Culicidae). AB - The purpose of this study was to investigate the repellent activities of essential oils from Cryptomeria japonica (sugi) against adults of mosquitoes Aedes aegypti and Aedes albopictus . Comparison of essential oils from four different plant parts of C. japonica revealed that essential oil from its leaf exhibited the best repellent activity against mosquitoes. To understand the relationship between volatile organic compounds and repellent activity, the solid phase microextraction (SPME) method was employed to analyze volatile organic compounds of leaf essential oil. The SPME fiber was coated with divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS). The major volatile organic compounds in the cage were 3-carene, alpha-terpinene, limonene, gamma terpinene, and terpinolene at 0 min. Results demonstrated that (-)-terpinen-4-ol was the major volatile organic compound adsorbed by SPME fiber during repellent assays. Furthermore, the repellent activities of six compounds against adults of the mosquitoes were evaluated, and the results revealed that (-)-terpinen-4-ol exhibited the best repellent activity against A. aegypti and A. albopictus. PMID- 19902949 TI - Finding the joker among the maize endogenous reference genes for genetically modified organism (GMO) detection. AB - The comparison of five real-time polymerase chain reaction (PCR) methods targeted at maize ( Zea mays ) endogenous sequences is reported. PCR targets were the alcohol dehydrogenase (adh) gene for three methods and high-mobility group (hmg) gene for the other two. The five real-time PCR methods have been checked under repeatability conditions at several dilution levels on both pooled DNA template from several genetically modified (GM) maize certified reference materials (CRMs) and single CRM DNA extracts. Slopes and R(2) coefficients of all of the curves obtained from the adopted regression model were compared within the same method and among all of the five methods, and the limit of detection and limit of quantitation were analyzed for each PCR system. Furthermore, method equivalency was evaluated on the basis of the ability to estimate the target haploid genome copy number at each concentration level. Results indicated that, among the five methods tested, one of the hmg-targeted PCR systems can be considered equivalent to the others but shows the best regression parameters and a higher repeteability along the dilution range. Thereby, it is proposed as a valid module to be coupled to different event-specific real-time PCR for maize genetically modified organism (GMO) quantitation. The resulting practicability improvement on the analytical control of GMOs is discussed. PMID- 19902950 TI - Markers of pesticide exposure in irrigated rice cultures. AB - The objective of this research is to verify the genotoxicity caused by pesticides used in irrigated rice cultures in Ararangua city in the southern Brazilian state of Santa Catarina through the alkaline comet assay in peripheral blood of Geophagus brasiliensis and to analyze the toxicity of the water using Daphnia magna as sentinel organism. Three collections of water and fish were made in the main rice ditch, and one collection for the control group was taken in the Ararangua River. The toxicity test with D. magna and the comet assay followed protocols previously described. The toxicity factor for the control group and collections 1, 2, and 3 were, respectively, 0, 1, 0, and 2. The comet assay demonstrated significant differences just in collection 2, in comparison to the control group and collections 1 and 3. These results, despite significant statistical data, are not a biological problem, because the values were not so large but serve to warn of a possible disruption of the balance in this environment system. PMID- 19902951 TI - Enhanced antioxidant and anti-inflammatory activities of Monascus pilosus fermented products by addition of turmeric to the medium. AB - Monascus sp. fermented products are known for their antihypercholesterolemic effects; however, their antioxidant and anti-inflammatory activities are different from those of many plant-derived foods. To evaluate the effect of turmeric addition into the medium on the antioxidant and anti-inflammatory activities of Monascus pilosus fermented products, we cultured uninoculated PDB medium (PDB), inoculated PDB medium (MP), uninoculated turmeric-containing PDB medium (PDBT), and inoculated turmeric-containing PDB medium (MPT). The broth and mycelia were collected, freeze-dried, and extracted to evaluate their free radical scavenging and iron-chelating activities, inhibition of peroxidation, phenolic and curcuminoid contents, and cellular antioxidant activity. The effects of the extracts on cell viability, cytokines and nitric oxide (NO) production, and expression of enzymes that regulate antioxidation and inflammation were also evaluated. The results showed that MPT had a significantly higher antioxidant activity than PDB, MP, and PDBT at all fermentation time points; moreover, the fermentation process significantly increased the phenolic and curcuminoid contents of MPT. As compared with MP, MPT had a more significant effect on down regulating the production of NO and TNF-alpha as well as the expression of inducible nitric oxide synthase, cyclooxygenase-2, glutathione peroxidase, superoxidase dismutase, and catalase. After the inherent levels of antioxidant and anti-inflammatory capacities were increased, the modified M. pilosus fermented product demonstrated a higher antiatherosclerotic value than the unmodified product. PMID- 19902952 TI - A selenide-based approach to photochemical cleavage of peptide and protein backbones at engineered backbone esters. AB - A strategy for photochemical cleavage of peptide and protein backbones is described, which is based on a selenide-mediated cleavage of a backbone ester moiety. Studies in model systems establish the viability of the chemistry and suggest the method could be a valuable tool for chemical biology studies of proteins. PMID- 19902954 TI - Discovery of 3-aryl-4-isoxazolecarboxamides as TGR5 receptor agonists. AB - A series of 3-aryl-4-isoxazolecarboxamides identified from a high-throughput screening campaign as novel, potent small molecule agonists of the human TGR5 G protein coupled receptor is described. Subsequent optimization resulted in the rapid identification of potent exemplars 6 and 7 which demonstrated improved GLP 1 secretion in vivo via an intracolonic dose coadministered with glucose challenge in a canine model. These novel TGR5 receptor agonists are potentially useful therapeutics for metabolic disorders such as type II diabetes and its associated complications. PMID- 19902956 TI - Revisiting the kinetics and mechanism of the tetrathionate-hypochlorous acid reaction in nearly neutral medium. AB - The tetrathionate-hypochlorous acid reaction has been investigated in nearly neutral medium at I = 0.5 M ionic strength and T = 25.0 +/- 0.1 degrees C in dihydrogen-phosphate-hydrogen-phosphate buffer by UV-vis spectrophotometry. In excess of hypochlorous acid, the stoichiometry was found to be S(4)O(6)(2-) + 7HOCl + 3H(2)O --> 4SO(4)(2-) + 7Cl(-) + 13H(+), but in excess of tetrathionate colloidal sulfur precipitates. On the basis of the simultaneous evaluation of the kinetic curves, a nine-step kinetic model with four fitted and five fixed rate coefficients is proposed. Analogous oxidation reactions of tetrathionate are also compared and discussed. PMID- 19902957 TI - Synthesis of 9-fluorenylidenes and 9,10-phenanthrenes through palladium-catalyzed aryne annulation by o-halostyrenes and o-halo allylic benzenes. AB - A number of functionally substituted 9-fluorenylidenes and 9,10-phenanthrenes have been synthesized from substituted o-halostyrenes and o-halo allylic benzenes respectively in good yields by the palladium-catalyzed annulation of arynes. The methodology tolerates a variety of functional groups, including cyano, ester, aldehyde, and ketone groups, occurs under relatively mild reaction conditions, and involves the generation of two new carbon-carbon bonds, thus providing these important carbocyclic ring systems in a single synthetic step. PMID- 19902958 TI - Solid phase synthesis of novel pyrrolidinedione analogs as potent HIV-1 integrase inhibitors. AB - A novel series of HIV-1 integrase inhibitors were identified from a 100 member (4R(1) x 5R(2) x 5R(3)) library of pyrrolidinedione amides. A solid-phase route was developed which facilitates the simultaneous variation at R(1), R(2), and R(3) of the pyrrolidinedione scaffold. The resulting library samples were assayed for HIV-1 integrase activity and analyzed to determine the R(1), R(2), and R(3) reagent contributions towards the activity. PMID- 19902959 TI - Ferric stability constants of representative marine siderophores: marinobactins, aquachelins, and petrobactin. AB - The coordination of iron(III) to the marine amphiphilic marinobactin and aquachelin siderophores, as well as to petrobactin, an unusual 3,4 dihydroxybenzoyl siderophore is reported. Potentiometric titrations were performed on the apo siderophore to determine the ligand pK(a) values, as well as the complex formed with addition of 1 equiv of Fe(III). The log K(ML) values for Fe(III)-marinobactin-E and Fe(III)-aquachelin-C are 31.80 and 31.4, respectively, consistent with the similar coordination environment in each complex, while log K(ML) for Fe(III)-petrobactin is estimated to be about 43. The pK(a) of the beta hydroxyaspartyl hydroxyl group was determined to be 10.8 by (1)H NMR titration. (13)C NMR and IR spectroscopy were used to investigate Ga(III) coordination to the marinobactins. The coordination-induced shifts (CIS) in the (13)C NMR spectrum of Ga(III)-marinobactin-C compared to apo-marinobactin-C indicates that the hydroxamate groups are coordinated to Ga(III); however, the lack of CISs for the carbons of the beta-hydroxyamide group suggests this moiety is not coordinated in the Ga(III) complex. Differences in the IR spectrum of Ga(III) marinobactin-C and Fe(III)-marinobactin-C in the 1600-1700 cm(-1) region also corroborates Fe(III) is coordinated to the beta-hydroxyamide moiety, whereas Ga(III) is not coordinated. PMID- 19902960 TI - The spin chemistry and magnetic resonance of H2@C60. From the Pauli principle to trapping a long lived nuclear excited spin state inside a buckyball. AB - One of the early triumphs of quantum mechanics was Heisenberg's prediction, based on the Pauli principle and wave function symmetry arguments, that the simplest molecule, H(2), should exist as two distinct species-allotropes of elemental hydrogen. One allotrope, termed para-H(2) (pH(2)), was predicted to be a lower energy species that could be visualized as rotating like a sphere and possessing antiparallel ( upward arrow downward arrow) nuclear spins; the other allotrope, termed ortho-H(2) (oH(2)), was predicted to be a higher energy state that could be visualized as rotating like a cartwheel and possessing parallel ( upward arrow upward arrow) nuclear spins. This remarkable prediction was confirmed by the early 1930s, and pH(2) and oH(2) were not only separated and characterized but were also found to be stable almost indefinitely in the absence of paramagnetic "spin catalysts", such as molecular oxygen, or traces of paramagnetic impurities, such as metal ions. The two allotropes of elemental hydrogen, pH(2) and oH(2), may be quantitatively incarcerated in C(60) to form endofullerene guest@host complexes, symbolized as pH(2)@C(60) and oH(2)@C(60), respectively. How does the subtle difference in nuclear spin manifest itself when hydrogen allotropes are incarcerated in a buckyball? Can the incarcerated "guests" communicate with the outside world and vice versa? Can a paramagnetic spin catalyst in the outside world cause the interconversion of the allotropes and thereby effect a chemical transformation inside a buckyball? How close are the measurable properties of H(2)@C(60) to those computed for the "quantum particle in a spherical box"? Are there any potential practical applications of this fascinating marriage of the simplest molecule, H(2), with one of the most beautiful of all molecules, C(60)? How can one address such questions theoretically and experimentally? A goal of our studies is to produce an understanding of how the H(2) guest molecules incarcerated in the host C(60) can "communicate" with the chemical world surrounding it. This world includes both the "walls" of the incarcerating host (the carbon atom "bricks" that compose the wall) and the "outside" world beyond the atoms of the host walls, namely, the solvent molecules and selected paramagnetic molecules added to the solvent that will have special spin interactions with the H(2) inside the complex. In this Account, we describe the temperature dependence of the equilibrium of the interconversion of oH(2)@C(60) and pH(2)@C(60) and show how elemental dioxygen, O(2), a ground-state triplet, is an excellent paramagnetic spin catalyst for this interconversion. We then describe an exploration of the spin spectroscopy and spin chemistry of H(2)@C(60). We find that H(2)@C(60) and its isotopic analogs, HD@C(60) and D(2)@C(60), provide a rich and fascinating platform on which to investigate spin spectroscopy and spin chemistry. Finally, we consider the potential extension of spin chemistry to another molecule with spin isomers, H(2)O, and the potential applications of the use of pH(2)@C(60) as a source of latent massive nuclear polarization. PMID- 19902961 TI - Organic light-emitting diodes on solution-processed graphene transparent electrodes. AB - Theoretical estimates indicate that graphene thin films can be used as transparent electrodes for thin-film devices such as solar cells and organic light-emitting diodes, with an unmatched combination of sheet resistance and transparency. We demonstrate organic light-emitting diodes with solution processed graphene thin film transparent conductive anodes. The graphene electrodes were deposited on quartz substrates by spin-coating of an aqueous dispersion of functionalized graphene, followed by a vacuum anneal step to reduce the sheet resistance. Small molecular weight organic materials and a metal cathode were directly deposited on the graphene anodes, resulting in devices with a performance comparable to control devices on indium-tin-oxide transparent anodes. The outcoupling efficiency of devices on graphene and indium-tin-oxide is nearly identical, in agreement with model predictions. PMID- 19902962 TI - Wafer-scale fabrication of separated carbon nanotube thin-film transistors for display applications. AB - Preseparated, semiconductive enriched carbon nanotubes hold great potential for thin-film transistors and display applications due to their high mobility, high percentage of semiconductive nanotubes, and room-temperature processing compatibility. Here in this paper, we report our progress on wafer-scale processing of separated nanotube thin-film transistors (SN-TFTs) for display applications, including key technology components such as wafer-scale assembly of high-density, uniform separated nanotube networks, high-yield fabrication of devices with superior performance, and demonstration of organic light-emitting diode (OLED) switching controlled by a SN-TFT. On the basis of separated nanotubes with 95% semiconductive nanotubes, we have achieved solution-based assembly of separated nanotube thin films on complete 3 in. Si/SiO(2) wafers, and further carried out wafer-scale fabrication to produce transistors with high yield (>98%), small sheet resistance ( approximately 25 kOmega/sq), high current density ( approximately 10 microA/microm), and superior mobility ( approximately 52 cm(2) V(-1) s(-1)). Moreover, on/off ratios of >10(4) are achieved in devices with channel length L > 20 microm. In addition, OLED control circuit has been demonstrated with the SN-TFT, and the modulation in the output light intensity exceeds 10(4). Our approach can be easily scaled to large areas and could serve as critical foundation for future nanotube-based display electronics. PMID- 19902963 TI - Electric field directed self-assembly of cuprous oxide nanostructures for photon sensing. AB - We demonstrate a novel chemical-free water-based technique to synthesize various forms of cuprous oxide nanostructures at room temperature. The self-assemblies of these nanostructures are formed by the anodic oxidation of Cu in deionized (DI) water. Direct growth of these nanostructures on SiO(2)/Si (100) substrate has been successfully achieved by tuning the bias voltage and the growth duration. A variety of nanostructures from one-dimensional nanowires to different complex two and three-dimensional structures are successfully grown by this method. We show that the morphological evolution in the self-assembly of the structures strongly depends on the spatial electric field distribution on the substrate. Furthermore, the electrical devices made from these nanowire networks exhibit promising photon sensing characteristics under white light illumination and can be exploited for future applications in photodetection and photovoltaic studies at the nanoscale level. PMID- 19902964 TI - Synthesis of biphenyl proteomimetics as estrogen receptor-alpha coactivator binding inhibitors. AB - A novel series of biphenyl proteomimetic compounds were designed as estrogen receptor-alpha (ER(alpha)) coactivator binding inhibitors. Synthesis was accomplished through a convergent approach, employing Suzuki coupling chemistry to ligate the individual modular units. Initial biological results support the ability of these compounds to compete for the ER(alpha) coactivator binding groove. PMID- 19902966 TI - Mechanism of fibril formation by a 39-residue peptide (PAPf39) from human prostatic acidic phosphatase. AB - PAPf39 is a 39-residue peptide fragment from the sequence of human prostatic acidic phosphatase. This peptide was shown to form amyloid-like fibrils, which have been implicated in facilitating semen-mediated HIV transmission. Thus understanding molecular details of PAPf39 peptide fibril formation may aid in elucidating the mechanism of how PAPf39 fibrils are involved in HIV etiology. To this end, the kinetics of PAPf39 peptide fibrillization was studied using a battery of biophysical methods (atomic force microscopy, ThT fluorescence assays, far-UV circular dichroism spectroscopy, deep-UV resonance Raman spectroscopy, size exclusion chromatography, analytical ultracentrifugation, and small-angle X ray scattering). It has been shown that fibril formation follows a nucleation dependent elongation mechanism. Several critical factors for fibrillization have been identified. It was shown that agitation and/or seeding is required for fibril formation at 37 degrees C and neutral pH, with an additional requirement of a salt concentration above approximately 100 mM. Fibril formation by the PAPf39 peptide is inhibited by low pH or by low salt concentration at neutral pH. These observations suggest that the nucleation and fibrillization of the PAPf39 peptide are a tug-of-war between the interactions formed upon agitation and the electrostatic interactions, modulated by pH and salt concentration. PMID- 19902967 TI - Structurally diverse limonoids from the fruits of Swietenia mahagoni. AB - Eleven new mexicanolide-type limonoids, swietmanins A-I (1-4, 7-11), 2-hydroxy-3 O-isobutyrylproceranolide (5), 2-hydroxy-3-O-benzoylproceranolide (6), and a new andirobin-type limonoid, swietmanin J (12), together with 19 known compounds were isolated from the fruits of Swietenia mahagoni. Their structures were established on the basis of spectroscopic methods. These compounds were evaluated against a small panel of microorganisms. PMID- 19902965 TI - Activating mutations in TOR are in similar structures as oncogenic mutations in PI3KCalpha. AB - TOR (Target of Rapamycin) is a highly conserved Ser/Thr kinase and a central controller of cell growth. Using the crystal structure of the related lipid kinase PI3KCgamma, we built a model of the catalytic region of TOR, from the FAT domain to near the end of the FATC domain. The model reveals that activating mutations in TOR, identified in yeast in a genetic selection for Rheb independence, correspond to hotspots for oncogenic mutations in PI3KCalpha. The activating mutations are in the catalytic domain (helices kalpha3, kalpha9, kalpha11) and the helical domain of TOR. Docking studies with small molecule inhibitors (PP242, NVP-BEZ235, and Ku-0063794) show that drugs currently in development utilize a novel pharmacophore space to achieve specificity. Thus, our model provides insight on the regulation of TOR and may be useful in the design of new anticancer drugs. PMID- 19902968 TI - Human P2Y(14) receptor agonists: truncation of the hexose moiety of uridine-5' diphosphoglucose and its replacement with alkyl and aryl groups. AB - Uridine-5'-diphosphoglucose (UDPG) activates the P2Y(14) receptor, a neuroimmune system GPCR. P2Y(14) receptor tolerates glucose substitution with small alkyl or aryl groups or its truncation to uridine 5'-diphosphate (UDP), a full agonist at the human P2Y(14) receptor expressed in HEK-293 cells. 2-Thiouracil derivatives displayed selectivity for activation of the human P2Y(14) vs the P2Y(6) receptor, such as 2-thio-UDP 4 (EC(50) = 1.92 nM at P2Y(14), 224-fold selectivity vs P2Y(6)) and its beta-propyloxy ester 18. EC(50) values of the beta-methyl ester of UDP and its 2-thio analogue were 2730 and 56 nM, respectively. beta-tert-Butyl ester of 4 was 11-fold more potent than UDPG, but beta-aryloxy or larger, branched beta-alkyl esters, such as cyclohexyl, were less potent. Ribose replacement of UDP with a rigid North or South methanocarba (bicyclo[3.1.0]hexane) group abolished P2Y(14) receptor agonist activity. alpha,beta-Methylene and difluoromethylene groups were well tolerated at the P2Y(14) receptor and are expected to provide enhanced stability in biological systems. alpha,beta-Methylene-2-thio-UDP 11 (EC(50) = 0.92 nM) was 2160-fold selective versus P2Y(6). Thus, these nucleotides and their congeners may serve as important pharmacological probes for the detection and characterization of the P2Y(14) receptor. PMID- 19902969 TI - Convergent synthesis of fostriecin via selective alkene couplings and regioselective asymmetric dihydroxylation. AB - A highly convergent synthesis of fostriecin is described, featuring sequential palladium-catalyzed Negishi cross couplings to form the C7-C8 bond and C8-methyl bond, followed by late-stage regio- and stereoselective dihydroxylation of C8-C9. PMID- 19902970 TI - The location of the biosurfactant surfactin in phospholipid bilayers supported on silica using neutron reflectometry. AB - We have investigated the formation of supported surfactin-phospholipid mixed bilayers using neutron reflectometry. Micellar mixtures of phospholipid (diphosphatidyl choline, DPPC), surfactin, and beta-d-dodecyl maltoside were used to make the deposition. When the surfactin concentration is at its critical micelle concentration (CMC = 6 x 10(-6) M) in the bulk solution, there is no adsorption at all on the silica. When the surfactin concentration is lowered below the CMC, a mixed bilayer of surfactin and DPPC is formed. Since surfactin does not adsorb on silica from solutions of surfactin alone, this shows that there is a strong attraction between surfactin and DPPC. The variation of adsorbed amount, composition, and structure of the adsorbed layer are consistent with the attractive interaction between surfactin and DPPC and with their respective negative and positive affinities for the silica surface. Three phospholipid isotopic contrasts were measured and used to define the composition and structure of the surfactin-phospholipid bilayer. The maximum amount of surfactin in the bilayer reaches a mole fraction of about 0.2 and this is located in the outer leaflet of the bilayer within the headgroup and part of the adjacent chain region. PMID- 19902971 TI - Association of phosphatidic acid with the bovine mitochondrial ADP/ATP carrier. AB - The beef heart adenine nucleotide carrier protein (Anc) of the inner mitochondrial membrane can be purified in a form stabilized by binding the inhibitor carboxyatractyloside. The protein is copurified with bound lipid. We show for the first time that phosphatidic acid, although a minor component, is one of the lipids bound to Anc. The short spin-lattice relaxation time found by (31)P magic angle spinning nuclear magnetic resonance (MAS/NMR) for phosphatidic acid indicates that it is tightly bound to the protein. However, this lipid also has a comparatively small chemical shift anisotropy, suggesting that it can undergo rapid reorientation in space. In contrast, most of the lipid bound to Anc shows anisotropic motion typical of a bilayer arrangement. The phosphatidic acid that is detected in the purified preparation of Anc is also shown to be present initially in the unfractionated mitochondria, prior to the isolation of Anc. In Triton-solubilized mitochondria, phosphatidic acid, cardiolipin, phosphatidylethanolamine, and phosphatidylcholine exhibit resonance lines in the static (31)P NMR spectra, but in the purified Anc, only the phosphatidylethanolamine and phosphatidylcholine can be detected by this method, even though the other lipids are still present. This demonstrates that the phosphatidic acid and cardiolipin are interacting with the Anc. The thermal denaturation of the Anc was determined by differential scanning calorimetry. The protein denatures at 74 degrees C both before and after the NMR studies with the same characteristics. PMID- 19902972 TI - Rectification of the ionic current through carbon nanotubes by electrostatic assembly of polyelectrolytes. AB - Rectification of the ionic current flowing through nanotubes embedded in a polymeric membrane is achieved by selective adsorption of polycations to the nanotubes' mouths. A one-dimensional model of ionic flux through a nanotube with charged entrance regions qualitatively describes current-voltage curves before and after polycation exposure; reversal potential measurements confirm that charge reversal takes place upon polycation adsorption. The inherent simply of this electrostatic approach makes it attractive in membrane and nanofluidic applications employing rectification. PMID- 19902973 TI - Selective inhibitors of bacterial phosphopantothenoylcysteine synthetase. AB - Bacterial phosphopantothenolycysteine synthetase (PPCS) catalyzes the formation of phosphopantothenoylcysteine (PPC) from (R)-phosphopantothenate, l-cysteine, and cytidine-5'-triphosphate (CTP) and has been shown to be essential for growth and survival. The reaction proceeds through a phosphopantothenoyl cytidylate, mixed anhydride intermediate. Both structural and kinetic characterization studies on PPCS have shown differences in the nucleobase binding site between the bacterial and human enzyme. We report for the first time the design and synthesis of mimics of the phosphopantothenoyl cytidylate, which proved to be potent inhibitors of PPCS. These compounds were evaluated in vitro against PPCS from human and several species of bacteria and showed marked selectivity (up to 1000 fold) toward the bacterial enzymes. A phosphodiester intermediate mimic was the most potent of the compounds synthesized and displayed slow-onset, tight-binding kinetics toward E. faecalis PPCS. PMID- 19902974 TI - The functional significance of lipid diversity: orientation of cholesterol in bilayers is determined by lipid species. AB - The chemical diversity of lipids and their complex arrangements in supramolecular assemblies are in stark contrast to our previous notions of them as passive structural components. For example, in plasma membranes, sphingolipids are primarily located in the outer monolayer, whereas unsaturated phospholipids are more abundant in the inner leaflet. Our recent results offer a direct contribution to the importance of lipid diversity in biological membranes. We have studied the location of cholesterol within polyunsaturated fatty acid (PUFA) bilayers doped with different amounts of monounsaturated (POPC) or disaturated (DMPC) lipids. Using deuterium labeling and neutron diffraction, we have found that in PUFA bilayers, cholesterol can be flipped from its known position in the bilayer center to its commonly assumed upright orientation simply by varying the amount of POPC. Although it takes approximately 50 mol % POPC to flip cholesterol in PUFA bilayers, the same effect is achieved with only 5 mol % DMPC, elegantly emphasizing cholesterol's affinity for saturated chains. It also suggests that the presence of PUFA in the inner leaflet of a cellular bilayer may enhance the transfer of cholesterol to the outer layer, potentially modifying raft composition and the local function of a membrane. PMID- 19902975 TI - Chiral amplification with a stereodynamic triaryl probe: assignment of the absolute configuration and enantiomeric excess of amino alcohols. AB - A stereodynamic, axially chiral triaryl probe is locked into a single conformation upon condensation with two amino alcohol substrates. The well defined chiral amplification in the diimines formed results in intense Cotton effects at high wavelengths that can be used for in situ CD analysis of the absolute configuration and ee of cyclic and acyclic substrates. PMID- 19902976 TI - Highly luminescent self-organized sub-2-nm EuOF nanowires. AB - Monodisperse sub-2-nm EuOF nanowires were obtained by manipulating the fluorophilicity between crystalline seeds and capping surfactant molecules during the thermolysis of Eu(CF(3)COO)(3) in oleic acid (OA) and oleylamine (OM). The uniform EuOF nanowires can self-organize on substrates to form parallel aligned superstructures and display strong Eu(3+)red emissions with high quantum yields of 65% under the UV light excitations due to the presence of dense surface Eu(3+) sites in the ultrathin nanowires as well as the passivation of the surface defects by the capping ligands. PMID- 19902977 TI - Breaking mechanism of single molecular junctions formed by octanedithiol molecules and Au electrodes. AB - We present a theoretical study of the elongation process of molecular junctions formed by octanedithiol molecule and Au electrodes. Five types of junctions that have different molecule-electrode coupling geometries are considered. It is found that the behavior of the H atom in the -SH group plays a crucial role in the system structure variation. The variation of the total energy and the average force needed to break the molecular junction are calculated, and each type of molecular junctions is found to have a characteristic breaking force. Comparing our theoretical results with those from experiment shows that the most probable coupling geometry was neglected in almost all the previous work. A dynamic analysis of the electronic structure of the molecular junctions is used to understand the variation of the system configuration. PMID- 19902978 TI - Shape control of CdSe nanocrystals with zinc blende structure. AB - Systematically shape-controlled synthesis of inorganic nanocrystals has attracted increasing attention recently for both fundamental and technological interest. The shape evolution of CdSe nanocrystals with wurtzite structure in a unified method has been previously well studied, while the study on shape evolution of CdSe nanocrystals with zinc blende structure still remains challenging. Here, we demonstrate a systematic shape variation of zinc blende CdSe nanocrystals in a modified organometallic approach, in which distinct shapes of cube-shaped, sphere shaped, tetrahedron-shaped, and branched CdSe nanocrystals with high yield and good uniformity are obtained. The crucial factor that influences the shape controlled process is the reactive temperature. This wide variation of shapes provides important information about the growth of CdSe nanocrystals, and it can also help in the shape-controlled synthesis of other nanocrystals that are bound with uniform crystal planes. PMID- 19902979 TI - Thermally switchable one- and two-dimensional arrays of single-walled carbon nanotubes in a polymeric system. AB - Fabrication of highly ordered arrays of single-walled carbon nanotubes (SWNTs) has been of great interest for a wide range of potential applications. Here, we report thermally switchable one- and two-dimensional arrays of individually isolated SWNTs formed by cooperative self-assembly of functionalized SWNTs and a block copolymer/water system. Small-angle X-ray scattering measurements reveal that when the block copolymer/water system is in an isotropic phase, two dimensional hexagonal arrays of SWNTs are formed by depletion attraction, and when the block copolymer/water system is in a lamellar phase, one-dimensional lattices of SWNTs intercalated in the polar regions of the polymeric lamellar structure are formed by entropically driven segregation and two-dimensional depletion attraction. These two SWNT arrays are thermally interchangeable, following the temperature-dependent phase behavior of the block copolymer/water system. PMID- 19902980 TI - Proteomic identification of overexpressed PRDX 1 and its clinical implications in ovarian carcinoma. AB - Ovarian carcinoma is the most lethal gynecological malignancy in worldwide. The discovery of reliable marker for early detection of ovarian carcinoma is critical for increasing patient's survival because high mortality rate is associated with late diagnosis of this tumor. In the present study, we performed comparative analysis of whole proteomes between serous borderline tumor and serous carcinoma to identify a useful biomarker for the early diagnosis and progression of ovarian carcinoma. Nine proteins were significantly overexpressed in ovarian serous carcinomas compared to borderline tumors. After validation with Western blotting and immunohistochemical analysis, prdx 1 was found to be the strongest overexpressed protein in malignant ovarian tumors among the selected proteins. In addition, the high level of prdx 1 expression (>50% positive cancer cells) was significantly correlated with poor overall survival in ovarian serous carcinomas. On a multivariate cox analysis, the relative risk of death was 8.74 in patients with serous carcinomas showing >50% of prdx 1-positive cancer cells. Our results suggest that prdx 1 may be a useful diagnostic and prognostic biomarker in ovarian carcinoma, especially serous carcinoma. PMID- 19902981 TI - Why do pedometers work?: a reflection upon the factors related to successfully increasing physical activity. AB - The results of two recent independent meta-analyses focused on pedometer-based programmes conclude that they work; that is, they are effective. Specifically, physical activity increases while blood pressure and weight decrease as a result of participating in a pedometer-based intervention. An improved understanding of the unique measurement and motivational properties of pedometers as behaviour change tools will assist researchers and practitioners to maximize benefits. In an effort to begin to outline why pedometers work, for whom, and under what conditions, the purpose of this current opinion article is to explore the published literature (drawing heavily from those studies previously identified in published meta-analyses and our own work in this area) to identify factors related to using pedometers to increase physical activity. In particular it is important to: (i) gain a better understanding of the activity-promoting characteristics of pedometers; (ii) determine effective elements of pedometer based programming; and (iii) identify participants who engage in, and benefit most from, such programming. Pedometers are most sensitive to walking behaviours, which is consistent with public health and clinical approaches to increasing physical activity. Specifically, they offer an affordable and accessible technology that is simplistic in output, low-literacy friendly, and immediately understandable to end-users. Support materials are becoming readily available for researchers and practitioners in terms of expected (normative or benchmark) values, patterns of change, indices to aid screening and interpretation, and measurement protocols. Pedometer-based programme theory is now being articulated and tested, and the critical elements necessary to shape a successful programme are becoming more clearly defined. More research is needed, however, to compare the effectiveness of self-selected individualized goals with tailored goals (based on a specified baseline characteristic, for example), standardized goals (e.g. percentage-based increments) and pre-set uniformly administered goals (i.e. a volume total of 10 000 steps/day or an incremental total of 2000 extra steps/day for everyone). Since most studies of pedometer-based programmes have been of relatively short duration, it is unknown to what extent observed changes are sustainable or whether it is possible to continue to accrue benefits over long-term adherence. Peer delivery of treatment has the potential for enabling wider and less costly dissemination, although this has not been directly evaluated. In addition, the majority of pedometer-based programme participants to date have been women, suggesting that more research is needed on men and how they react to this form of physical activity intervention. Increases in steps/day have been negatively correlated with baseline values, indicating that those with lower baseline steps/day stand to make the greatest relative incremental increases in physical activity behaviour. A clearly articulated programme theory is lacking in most interventions. A clearer understanding is needed of what programme features, including the nature of goal-setting, are necessary for optimal participant success. Additionally, we need a better profile of the participant who benefits most, and/or requires additional or alternative strategies to succeed in their personal behaviour-change attempts. Continued efforts to refine answers regarding what works well for whom under what conditions will foster evidence-based applications of pedometer-based programmes. PMID- 19902982 TI - Encouraging walking for transport and physical activity in children and adolescents: how important is the built environment? AB - In the post-World War II era, there have been dramatic changes to the environment that appear to be having a detrimental impact on the lifestyles and incidental physical activities of young people. These changes are not trivial and have the potential to influence not only physical health, but also mental health and child development. However, the evidence of the impact of the built environment on physical activity to date is inconsistent. This review examines the evidence on the association between the built environment and walking for transport as well as physical activity generally, with a focus on methodological issues that may explain inconsistencies in the literature to date. It appears that many studies fail to measure behaviour-specific environmental correlates, and insufficient attention is being given to differences according to the age of study participants. Higher levels of out-of-school-hours physical activity and walking appear to be significantly associated with higher levels of urban density and neighbourhoods with mixed-use planning, especially for older children and adolescents. Proximate recreational facilities also appear to predict young people's level of physical activity. However, there are inconsistencies in the literature involving studies with younger children. Independent mobility increases with age. For younger children, the impact of the built environment is influenced by the decision-making of parents as the gatekeepers of their behaviour. Cross-cultural differences may also be present and are worthy of greater exploration. As children develop and are given more independent mobility, it appears that the way neighbourhoods are designed - particularly in terms of proximity and connectivity to local destinations, including schools and shopping centres, and the presence of footpaths - becomes a determinant of whether children are able, and are permitted by their parents, to walk and use destinations locally. If older children and adolescents are to enjoy health and developmental benefits of independent mobility, a key priority must be in reducing exposure to traffic and in increasing surveillance on streets (i.e. 'eyes-on-the-street') through neighbourhood and building design, by encouraging others to walk locally, and by discouraging motor vehicle use in favour of walking and cycling. Parents need to be assured that the rights and safety of pedestrians (and cyclists) - particularly child pedestrians and cyclists - are paramount if we are to turn around our 'child-free streets', now so prevalent in contemporary Australian and US cities. There remains a need for more age- and sex specific research using behaviour- and context-specific measures, with a view to building a more consistent evidence base to inform future environmental interventions. PMID- 19902983 TI - Does antioxidant vitamin supplementation protect against muscle damage? AB - The high forces undergone during repetitive eccentric, or lengthening, contractions place skeletal muscle under considerable stress, in particular if unaccustomed. Although muscle is highly adaptive, the responses to stress may not be optimally regulated by the body. Reactive oxygen species (ROS) are one component of the stress response that may contribute to muscle damage after eccentric exercise. Antioxidants may in turn scavenge ROS, thereby preventing or attenuating muscle damage. The antioxidant vitamins C (ascorbic acid) and E (tocopherol) are among the most commonly used sport supplements, and are often taken in large doses by athletes and other sportspersons because of their potential protective effect against muscle damage. This review assesses studies that have investigated the effects of these two antioxidants, alone or in combination, on muscle damage and oxidative stress. Studies have used a variety of supplementation strategies, with variations in dosage, timing and duration of supplementation. Although there is some evidence to show that both antioxidants can reduce indices of oxidative stress, there is little evidence to support a role for vitamin C and/or vitamin E in protecting against muscle damage. Indeed, antioxidant supplementation may actually interfere with the cellular signalling functions of ROS, thereby adversely affecting muscle performance. Furthermore, recent studies have cast doubt on the benign effects of long-term, high-dosage antioxidant supplementation. High doses of vitamin E, in particular, may increase all-cause mortality. Although some equivocation remains in the extant literature regarding the beneficial effects of antioxidant vitamin supplementation on muscle damage, there is little evidence to support such a role. Since the potential for long-term harm does exist, the casual use of high doses of antioxidants by athletes and others should perhaps be curtailed. PMID- 19902984 TI - Bovine colostrum supplementation and exercise performance: potential mechanisms. AB - Bovine colostrum (BC) is rich in immune, growth and antimicrobial factors, which promote tissue growth and the development of the digestive tract and immune function in neonatal calves. Although the value of BC to human adults is not well understood, supplementation with BC is becoming increasingly popular in trained athletes to promote exercise performance. The combined presence of insulin-like growth factors (IGF), transforming growth factors, immunoglobulins, cytokines, lactoferrin and lysozyme, in addition to hormones such as growth hormone, gonadotrophin-releasing hormone, luteinizing hormone-releasing hormone and glucocorticoids, would suggest that BC might improve immune function, gastrointestinal integrity and the neuroendocrine system, parameters that may be compromised as a result of intensive training. A review of studies investigating the influence of BC supplementation on exercise performance suggests that BC supplementation is most effective during periods of high-intensity training and recovery from high-intensity training, possibly as a result of increased plasma IGF-1, improved intramuscular buffering capacity, increases in lean body mass and increases in salivary IgA. However, there are contradicting data for most parameters that have been considered to date, suggesting that small improvements across a range of parameters might contribute to improved performance and recovery, although this cannot be concluded with certainty because the various doses and length of supplementation with BC in different studies prevent direct comparison of results. Future research on the influence of BC on sports performance will only be of value if the dose and length of supplementation of a well-defined BC product is standardized across studies, and the bioavailability of the active constituents in BC is determined. PMID- 19902985 TI - Oligomenorrhoea in exercising women: a polycystic ovarian syndrome phenotype or distinct entity? AB - To date, the predominant mechanism underlying menstrual disturbances in exercising women supports an underlying energy deficiency-related aetiology, in which a failure to compensate dietary intake for the energy cost of exercise suppresses reproductive function. Increasing evidence demonstrates that energy deficiency plays a causal role in the induction of amenorrhoea in exercising women, and consistent with this mechanism are findings of glucoregulatory perturbations such as low triiodothyronine, reduced insulin secretion and elevated cortisol, growth hormone and ghrelin levels. The menstrual disturbance that may differ in its energetic characteristics and, perhaps in its androgenic and ovarian steroid environment, is oligomenorrhoea. We conducted a systematic review of the literature to begin to understand whether oligomenorrhoea in exercising women is a mild subclinical phenotype of polycystic ovarian syndrome (PCOS) in which exercise is conferring beneficial effects in protecting women from the classic PCOS phenotype, or whether oligomenorrhoea is part of the spectrum of menstrual disturbances caused by an energy deficiency that is often reported in exercising women with menstrual disturbances. We included observational, randomized controlled trials and cross-sectional studies that reported clinical, hormonal and metabolic profiles in exercising women with amenorrhoea or oligomenorrhoea and in women with PCOS. Previous studies examining the underlying mechanisms and consequences of exercise-associated menstrual disturbances have grouped exercising amenorrhoeic and oligomenorrhoeic women into a single group, and have relied primarily on self-reported menstrual history. Although scarce, the data available to date suggest that hyperandrogenism, such as that observed in PCOS, may likely be associated with oligomenorrhoea in exercising women, and may not always represent hypothalamic inhibition secondary to an energy deficiency. It is critical to closely examine the metabolic and endocrine status of women with menstrual disturbances because the treatment strategies for energy deficiency-related menstrual disturbances differ from that of disturbances traceable to hyperandrogenaemia. Further investigation is necessary to explore whether different endocrine aetiologies underly menstrual disturbances, particularly oligomenorrhoea, in physically active women. PMID- 19902988 TI - Tacrolimus population pharmacokinetic-pharmacogenetic analysis and Bayesian estimation in renal transplant recipients. AB - OBJECTIVES: The aims of this study were (i) to investigate the population pharmacokinetics of tacrolimus in renal transplant recipients, including the influence of biological and pharmacogenetic covariates; and (ii) to develop a Bayesian estimator able to reliably estimate the individual pharmacokinetic parameters and inter-dose area under the blood concentration-time curve (AUC) from 0 to 12 hours (AUC(12)) in renal transplant patients. METHODS: Full pharmacokinetic profiles were obtained from 32 renal transplant patients at weeks 1 and 2, and at months 1, 3 and 6 post-transplantation. The population pharmacokinetic analysis was performed using the nonlinear mixed-effect modelling software NONMEM version VI. Patients' genotypes were characterized by allelic discrimination for PXR -25385C>T genes. RESULTS: Tacrolimus pharmacokinetics were well described by a two-compartment model combined with an Erlang distribution to describe the absorption phase, with low additive and proportional residual errors of 1.6 ng/mL and 9%, respectively. Both the haematocrit and PXR -25385C>T single nucleotide polymorphism (SNP) were identified as significant covariates for apparent oral clearance (CL/F) of tacrolimus, which allowed improvement of prediction accuracy. Specifically, CL/F decreased gradually with the number of mutated alleles for the PXR -25385C>T SNP and was inversely proportional to the haematocrit value. However, clinical criteria of relevance, mainly the decrease in interindividual variability and residual error, led us to retain only the haematocrit in the final model. Maximum a posteriori Bayesian forecasting allowed accurate prediction of the tacrolimus AUC(12) using only three sampling times (at 0 hour [predose] and at 1 and 3 hours postdose) in addition to the haematocrit value, with a nonsignificant mean AUC bias of 2% and good precision (relative mean square error = 11%). CONCLUSION: Population pharmacokinetic analysis of tacrolimus in renal transplant recipients showed a significant influence of the haematocrit on its CL/F and led to the development of a Bayesian estimator compatible with clinical practice and able to accurately predict tacrolimus individual pharmacokinetic parameters and the AUC(12). PMID- 19902989 TI - Population pharmacokinetic modelling of filgrastim in healthy adults following intravenous and subcutaneous administrations. AB - BACKGROUND AND OBJECTIVE: Filgrastim is a human granulocyte-colony stimulating factor (G-CSF). The biological activity of filgrastim is identical to that of endogenous G-CSF. It controls neutrophil production within the bone marrow by stimulating the proliferation, differentiation and survival of myeloid progenitor cells and some end-cell function activation. The purpose of this work is to propose a target-mediated drug disposition pharmacokinetic model of filgrastim. METHODS: A mechanism-based population pharmacokinetic model was developed to account for receptor-mediated endocytosis as a mechanism for nonlinear disposition of G-CSF. Time profiles of serum filgrastim concentrations following subcutaneous doses of 2.5, 5 and 10 microg/kg and intravenous infusion of 5 microg/kg over 0.5 hour were studied. The pharmacokinetic model included first order elimination from the serum, receptor binding, turnover of free receptors and internalization of drug-receptor complexes. The proposed target-mediated drug disposition models served as a tool to study drug absorption and the impact of receptor binding on filgrastim clearance. RESULTS: Filgrastim was found to exhibit parallel absorption with first- and zero-order kinetics and bioavailability of 69.1%. The majority of the drug (58.6%) was absorbed by zero order processes, presumably through the lymphatic system. The equilibrium dissociation constant (K(d)) was estimated as 16.38 pM. CONCLUSION: The proposed model predicts that clearance is initially mostly governed by the binding of filgrastim to G-CSF receptors. Subsequently, the clearance slows down because of the saturation of binding sites, and occurs mostly via the linear (renal) pathway. Finally, for G-CSF concentrations lower than the K(d), target-mediated clearance dominates. The presented receptor-mediated model adequately describes filgrastim serum concentrations and quantifies the role of receptor binding in G CSF clearance. PMID- 19902990 TI - A case of severe cefepime-related neutropenia in a 15-year-old patient. AB - We describe the case of a 15-y-old boy with a post-surgical osteoarticular infection, who developed a severe neutropenia after 24 days of treatment with cefepime. Our report suggests that, although rare, severe neutropenia should be considered by clinicians when prescribing cefepime, especially if long-term therapy is expected. PMID- 19902991 TI - Methicillin-resistant Staphylococcus aureus from pressure ulcers in patients receiving home care in Palermo, Italy. PMID- 19902992 TI - Use of rapid diagnostic tests and choice of antibiotics in respiratory tract infections in primary healthcare--a 6-y follow-up study. AB - The aim of this retrospective study of electronic patient records in primary health care in Kalmar County, Sweden, was to describe consultations for respiratory tract infections (RTIs) in relation to age, choice of antibiotics and the use of rapid diagnostic tests. During the period 1999-2005, 240,445 visits for RTI were recorded. Children aged <2 y and especially those aged 2-16 y with acute otitis media (AOM), showed decreasing consultations between 2000 and 2005. The consultations for sore throat declined during the study period in all age groups and in 65% of these, antibiotics were prescribed, primarily penicillin V (82%). In sore throat, a positive Strep-A test result was followed by antibiotic prescription in about 92% of cases; when negative, the antibiotic prescription rate was 40%. C-reactive protein (CRP) was analyzed in 36% of all consultations for RTI. In common cold and acute bronchitis, the prescription rates of antibiotics rose with rising CRP. The results show that near-patient tests were used extensively, but often not in accordance with the guidelines. Antibiotic use decreased mainly as a consequence of declined visiting frequencies. This indicates that the new guidelines for AOM and sore throat may have influenced patient consultation habits more than physician prescribing habits. PMID- 19902993 TI - Typing of Candida isolates from patients with invasive infection and concomitant colonization. AB - We investigated the relationship between colonizing and invasive isolates from patients with candidaemia. Molecular typing was performed using random amplification of polymorphic DNA (RAPD) and multilocus sequence typing (MLST). We found MLST to be sufficient for typing Candida isolates, and that surveillance cultures are helpful in predicting concomitant invasive isolates, but not necessarily the pathogen involved in subsequent episodes. PMID- 19902995 TI - Regen: the industry responsible for cell-based therapies. PMID- 19902994 TI - Economic aspects of mandibular third molar surgery. AB - OBJECTIVE: The aim of this study was to make estimates from a dental care and societal perspective on costs of mandibular third molar surgery. MATERIAL AND METHODS: A total of 64 patients were recruited from three Swedish oral and maxillofacial specialist clinics. Calculations were made prospectively on utilization of labor time, specific medical services and materials, and standardized utilization of other direct costs. Indirect costs were identified from patient surveys. RESULTS: The base case average direct cost of surgery was 217 Euro. Adding the patient's average cost due to absence from work and transportation of 333 Euro increased overall costs to 550 Euro per patient. About 86% of the patients reported some absence following surgery. CONCLUSIONS: The indirect costs were on average higher than the direct costs, i.e. the patient's loss of time caused higher costs than the intervention per se. Appropriate indications for mandibular third molar removal can minimize the risks of complications and individual or societal costs. PMID- 19902996 TI - Pig induced pluripotent stem cells: a new resource for generating genetically modified pigs. PMID- 19902997 TI - Of mice and men: skin cells, stem cells and ethical uncertainties. PMID- 19902987 TI - Polymorphism of human cytochrome P450 2D6 and its clinical significance: part II. AB - Part I of this article discussed the potential functional importance of genetic mutations and alleles of the human cytochrome P450 2D6 (CYP2D6) gene. The impact of CYP2D6 polymorphisms on the clearance of and response to a series of cardiovascular drugs was addressed. Since CYP2D6 plays a major role in the metabolism of a large number of other drugs, Part II of the article highlights the impact of CYP2D6 polymorphisms on the response to other groups of clinically used drugs. Although clinical studies have observed a gene-dose effect for some tricyclic antidepressants, it is difficult to establish clear relationships of their pharmacokinetics and pharmacodynamic parameters to genetic variations of CYP2D6; therefore, dosage adjustment based on the CYP2D6 phenotype cannot be recommended at present. There is initial evidence for a gene-dose effect on commonly used selective serotonin reuptake inhibitors (SSRIs), but data on the effect of the CYP2D6 genotype/phenotype on the response to SSRIs and their adverse effects are scanty. Therefore, recommendations for dose adjustment of prescribed SSRIs based on the CYP2D6 genotype/phenotype may be premature. A number of clinical studies have indicated that there are significant relationships between the CYP2D6 genotype and steady-state concentrations of perphenazine, zuclopenthixol, risperidone and haloperidol. However, findings on the relationships between the CYP2D6 genotype and parkinsonism or tardive dyskinesia treatment with traditional antipsychotics are conflicting, probably because of small sample size, inclusion of antipsychotics with variable CYP2D6 metabolism, and co-medication. CYP2D6 phenotyping and genotyping appear to be useful in predicting steady-state concentrations of some classical antipsychotic drugs, but their usefulness in predicting clinical effects must be explored. Therapeutic drug monitoring has been strongly recommended for many antipsychotics, including haloperidol, chlorpromazine, fluphenazine, perphenazine, risperidone and thioridazine, which are all metabolized by CYP2D6. It is possible to merge therapeutic drug monitoring and pharmacogenetic testing for CYP2D6 into clinical practice. There is a clear gene-dose effect on the formation of O-demethylated metabolites from multiple opioids, but the clinical significance of this may be minimal, as the analgesic effect is not altered in poor metabolizers (PMs). Genetically caused inactivity of CYP2D6 renders codeine ineffective owing to lack of morphine formation, decreases the efficacy of tramadol owing to reduced formation of the active O-desmethyl-tramadol and reduces the clearance of methadone. Genetically precipitated drug interactions might render a standard opioid dose toxic. Because of the important role of CYP2D6 in tamoxifen metabolism and activation, PMs are likely to exhibit therapeutic failure, and ultrarapid metabolizers (UMs) are likely to experience adverse effects and toxicities. There is a clear gene-concentration effect for the formation of endoxifen and 4-OH-tamoxifen. Tamoxifen-treated cancer patients carrying CYP2D6*4, *5, *10, or *41 associated with significantly decreased formation of antiestrogenic metabolites had significantly more recurrences of breast cancer and shorter relapse-free periods. Many studies have identified the genetic CYP2D6 status as an independent predictor of the outcome of tamoxifen treatment in women with breast cancer, but others have not observed this relationship. Thus, more favourable tamoxifen treatment seems to be feasible through a priori genetic assessment of CYP2D6, and proper dose adjustment may be needed when the CYP2D6 genotype is determined in a patient. Dolasetron, ondansetron and tropisetron, all in part metabolized by CYP2D6, are less effective in UMs than in other patients. Overall, there is a strong gene concentration relationship only for tropisetron. CYP2D6 genotype screening prior to antiemetic treatment may allow for modification of antiemetic dosing. An alternative is to use a serotonin agent that is metabolized independently of CYP2D6, such as granisetron, which would obviate the need for genotyping and may lead to an improved drug response. To date, the functional impact of most CYP2D6 alleles has not been systematically assessed for most clinically important drugs that are mainly metabolized by CYP2D6, though some initial evidence has been identified for a very limited number of drugs. The majority of reported in vivo pharmacogenetic data on CYP2D6 are from single-dose and steady-state pharmacokinetic studies of a small number of drugs. Pharmacodynamic data on CYP2D6 polymorphisms are scanty for most drug studies. Given that genotype testing for CYP2D6 is not routinely performed in clinical practice and there is uncertainty regarding genotype-phenotype, gene-concentration and gene-dose relationships, further prospective studies on the clinical impact of CYP2D6 dependent metabolism of drugs are warranted in large cohorts. PMID- 19903001 TI - Plasticity of stem cells derived from adult periodontal ligament. AB - BACKGROUND: The neural crest contains pluripotent cells that can give rise to neurons and glial cells of the peripheral nervous system, endocrine cells, connective tissue cells, muscle cells and pigment cells during embryonic development. Stem cells derived from the neural crest may still reside in neural crest derivatives including the periodontal ligament (PDL). However, the pluripotency of PDL-derived stem cells has not been investigated. AIM: To identify subpopulations of stem cells from the adult PDL and study their pluripotency. Human PDLs were harvested from impacted wisdom teeth (patients aged 19-22 years). RESULTS: This study demonstrated that subpopulations of PDL cells expressed embryonic stem cell markers (Oct4, Sox2, Nanog and Klf4) and a subset of neural crest markers (Nestin, Slug, p75 and Sox10). Such PDL cell subpopulations exhibited the potential to differentiate into neurogenic, cardiomyogenic, chondrogenic and osteogenic lineages. Furthermore, preliminary evidence suggesting insulin production of PDL cells might be indicative of the generation of cells of the endodermal lineage. CONCLUSION: These findings suggest that the PDL may contain pluripotent stem cells that originate from the neural crest. Our observations open the door to prospective autologous therapeutic applications for a variety of conditions. PMID- 19903002 TI - Cardiac stem cell genetic engineering using the alphaMHC promoter. AB - AIMS: Cardiac stem cells (CSCs) show potential as a cellular therapeutic approach to blunt tissue damage and facilitate reparative and regenerative processes after myocardial infarction. Despite multiple published reports of improvement, functional benefits remain modest using normal stem cells delivered by adoptive transfer into damaged myocardium. The goal of this study is to enhance survival and proliferation of CSCs that have undergone lineage commitment in early phases as evidenced by expression of proteins driven by the alpha-myosin heavy chain (alphaMHC) promoter. The early increased expression of survival kinases augments expansion of the cardiogenic CSC pool and subsequent daughter progeny. MATERIALS & METHODS: Normal CSCs engineered with fluorescent reporter protein constructs under control of the alphaMHC promoter show transgene protein expression, confirming activity of the promoter in CSCs. Cultured CSCs from both nontransgenic and cardiac-specific transgenic mice expressing survival kinases driven by the alphaMHC promoter were analyzed to characterize transgene expression following treatments to promote differentiation in culture. RESULTS & CONCLUSION: Therapeutic genes controlled by the alphaMHC promoter can be engineered into and expressed in CSCs and cardiomyocyte progeny with the goal of improving the efficacy of cardiac stem cell therapy. PMID- 19903003 TI - Assessing the value of autologous and allogeneic cells for regenerative medicine. AB - The advantages and disadvantages of autologous and allogeneic human cells for regenerative medicine are summarized. The comparison of relative advantages includes: ease and cost of treating large numbers of patients, the speed of availability of therapy and the differing complexity of the development pathways. The comparison of relative disadvantages deals with issues such as variability of source material, the risks of cell abnormality and of viral and prion contamination, and the sensitive issues surrounding use of embryo-derived cells. From the comparisons, several potentially decisive issues are drawn out, such as possible immune response and teratoma formation, the impact of patents and the virtues of hospital versus industry-centered development. PMID- 19903004 TI - Using stem cells to mend the retina in ocular disease. AB - Retinal degenerative diseases are the leading cause of incurable blindness worldwide. Furthermore, existing pharmacological and surgical interventions are only partially effective in halting disease progression, thus adjunctive neuroprotective strategies are desperately needed to preserve vision. Stem cells appear to possess inherent neuroprotective abilities, at least in part by providing neurotrophic support to injured neurons. Advances in stem cell biology offer the hope of new therapies for a broad range of neurodegenerative conditions, including those of the retina. Experimental cell-mediated therapies also hint at the tantalizing possibility of achieving retinal neuronal replacement and regeneration, once cells are lost to the disease process. This article summarizes the latest advances in cell therapies for neuroprotection and regeneration in neurodegenerative pathologies of both the inner and outer retina. PMID- 19903005 TI - Engineering the CNS stem cell microenvironment. AB - The loss of neural tissue underlies the symptomatology of several neurological insults of disparate etiology, including trauma, cerebrovascular insult and neurodegenerative disease. Restoration of damaged neural tissue through the use of exogenous or endogenous neural stem or progenitor cells is an enticing therapeutic option provided one can control their proliferation, migration and differentiation. Initial attempts at CNS tissue engineering relied on the intrinsic cellular properties of progenitor cells; however, it is now appreciated that the microenvironment surrounding the cells plays an indispensible role in regulating stem cell behavior. This article focuses on attempts to engineer the neural stem cell microenvironment by utilizing the major cellular components of the niche (endothelial cells, astrocytes and ependymal cells) and the extracellular matrix in which they are embedded. PMID- 19903007 TI - Human embryonic stem cells and genomic instability. AB - Owing to their original properties, pluripotent human embryonic stem cells (hESCs) and their progenies are highly valuable not only for regenerative medicine, but also as tools to study development and pathologies or as cellular substrates to screen and test new drugs. However, ensuring their genomic integrity is one important prerequisite for both research and therapeutic applications. Until recently, several studies about the genomic stability of cultured hESCs had described chromosomal or else large genomic alterations detectable with conventional karyotypic methods. In the past year, several laboratories have reported many small genomic alterations, in the megabase-sized range, using more sensitive karyotyping methods, showing that hESCs are prone to acquire focal genomic abnormalities in culture. As these alterations were found to be nonrandom, these findings strongly advocate for high-resolution monitoring of human pluripotent stem cell lines, especially when intended to be used for clinical applications. PMID- 19903008 TI - Stem cells and regenerative medicine on the Asian horizon: an economic, industry and social perspective. AB - For the past decade, forays into stem cell research and regenerative medicine by institutes and companies based in the Asia-Pacific region have attracted global attention at levels unprecedented in the life sciences. The unique combination of economic pressures, competitiveness and opportunism, laissez-faire regulation, burgeoning investment in the life sciences and rapidly growing markets, coupled with its great diversity, have propelled the region to surge forward in some areas, but to stumble in others. This article provides a historical and scientific context to the state of stem cell research and clinical applications in the region, and highlights trends and new possibilities to watch for on the Asian horizon. PMID- 19903010 TI - Topical tacrolimus for the treatment of localized, idiopathic, newly diagnosed pyoderma gangrenosum. AB - Pyoderma gangrenosum (PG) is a rare neutrophilic dermatosis which may present in a classic ulcerative form or in atypical bullous, vegetative or pustular variants. It can be associated with several disorders or be idiopathic. Although systemic immunosuppressants remain the choice therapy for most cases of PG, a local approach should be considered in localized disease. Recently, topical tacrolimus has successfully been used as an off-label drug in localized PG. In the present study, five patients with localized, idiopathic, newly diagnosed PG were treated with topical tacrolimus monotherapy. Localized PG was defined as disease involving no more than 5% of the body surface area and presenting with no more than three lesions. Cultures performed on PG lesions both before and during tacrolimus treatment were negative. In all five patients complete remission was achieved within a mean time of 6 weeks and no relapses occurred; in three cases, tacrolimus was discontinued, while the remaining two patients were applying the drug as maintenance therapy at the time of writing. Thus, we suggest that topical tacrolimus monotherapy could represent the first-line treatment for PG that fulfils the following criteria: localized disease, idiopathic form and recent onset with negative microbiological tests on PG lesions. PMID- 19903011 TI - Glucagon receptor mediates calcium signaling by coupling to G alpha q/11 and G alpha i/o in HEK293 cells. AB - Glucagon induces intracellular Ca(2+) ([Ca(2+)](i)) elevation by stimulating glucagon receptor (GCGR). Such [Ca(2+)](i) signaling plays important physiological roles, including glycogenolysis and glycolysis in liver cells and the survival of beta-cells. Previous studies indicated that phospholipase C (PLC) might be involved in glucagon-mediated [Ca(2+)](i) response. Other studies also debated whether cAMP accumulation mediated by GCGR/G alpha(s) coupling contributes to [Ca(2+)](i) elevation. But the exact mechanisms remain uncertain. In the present study, we found that glucagon induces [Ca(2+)](i) elevation in HEK293 cells expressing GCGR. Removing extracellular Ca(2+) did not affect glucagon-stimulated [Ca(2+)](i) response. But depleting the intracellular Ca(2+) store by thapsigargin completely inhibited glucagon-induced [Ca(2+)](i) response. Experiments with forskolin and adenylyl cyclase inhibitor revealed that cAMP is not the cause of [Ca(2+)](i) response. Further studies with G alpha(q/11) RNAi and pertussis toxin (PTX) indicated that both G alpha(q/11) and G alpha(i/o) are involved. Combination of G alpha(q/11) RNAi and G alpha(i/o) inhibition almost completely abolished glucagon-induced [Ca(2+)](i) signaling. PMID- 19903006 TI - Diverse roles of the vasculature within the neural stem cell niche. AB - An interdependent relationship between the vascular and nervous systems begins during the earliest stages of development and persists through the mammalian lifespan. Accordingly, the process of adult neurogenesis involves the coordinated response of both systems to maintain a specialized microenvironment (niche) that tips the scale towards maintenance or regeneration, as needed. Understanding the nature and regulation of this balance will provide a foundation on which the potential for molecular- and stem cell-based therapies can be developed to treat prevalent CNS diseases and disorders. The vasculature is cited as a prominent feature within the adult subventricular zone and subgranular zone, known adult neural stem cell niches, helping to retain neural stem and progenitor cell potential. The vascular compartment within the neural stem cell niche has the unique opportunity to not only regulate neural stem and progenitor cells through direct contact with, and paracrine signaling from, endothelial and mural cells that make up blood vessels, but also integrates systemic signals into the local microenvironment via distribution of soluble factors from blood circulation to regulate stem cell niche behavior. Understanding the intricate role that the vasculature plays to influence neural stem cells in the context of niche regulation will help to bridge the gap from bench to bedside for the development of regeneration-based therapies for the CNS. PMID- 19903012 TI - Evaluation of soluble fibrin and D-dimer in the diagnosis of postoperative deep vein thrombosis. AB - Soluble fibrin (SF) and D-dimer are useful for making the diagnosis of deep vein thrombosis (DVT). However, the evidence for using such markers and optimal timing to diagnose postoperative DVT are unclear. We evaluate the usefulness of SF and D dimer testing for the diagnosis of postoperative DVT. A total of 207 patients who had total hip arthroplasty or knee arthroplasty were evaluated. SF and D-dimer were tested on postoperative days 1 and 7. DVT was confirmed with ultrasonography. SF level on postoperative day 1 was the most useful, although D dimer evaluation on postoperative days 1 and 7 was also useful. Using a SF cut off of more than 4.00 microg ml(-1), the sensitivity was 90%, the specificity was 33%. Although the SF and D-dimer tests cannot be used as stand-alone tests, SF and D-dimer are valuable screening tools. We recommend two-stage screening including first with the SF or D-dimer test, followed by ultrasonography or venography. PMID- 19903013 TI - Pharmacokinetics of PSC 833 (valspodar) in its Cremophor EL formulation in rat. AB - Valspodar is a P-glycoprotein inhibitor widely used in preclinical and clinical studies for overcoming multidrug resistance. Despite this, the pharmacokinetics of valspodar in rat, a commonly used animal model, have not been reported. Here, we report on the pharmacokinetics of valspodar in Sprague-Dawley rats following intravenous and oral administration of its Cremophor EL formulation, which has been used for humans in clinical trials. After intravenous doses, valspodar displayed properties of slow clearance and a large volume of distribution. Its plasma unbound fraction was around 15% in the Cremophor EL formulation used in the study. After 10 mg kg(-1) orally it was rapidly absorbed with an average maximal plasma concentration of 1.48 mg l(-1) within approximately 2 h. The mean bioavailability of valspodar was 42.8%. In rat, valspodar showed properties of low hepatic extraction and wide distribution, similar to that of its structural analogue cyclosporine A. PMID- 19903017 TI - Cell cycle aberrations in Alzheimer's disease: a novel therapeutic opportunity. PMID- 19903019 TI - Activated astrocytes: a therapeutic target in Alzheimer's disease? AB - Astrocytes become activated in Alzheimer's disease, contributing to and reinforcing an inflammatory cascade. A large body of evidence suggests that by transforming from a basal to a reactive state, astrocytes neglect their neurosupportive functions, thus rendering neurons vulnerable to neurotoxins, including proinflammatory cytokines and reactive oxygen species. This review highlights three important astrocytic functions that may be impaired in neurodegenerative diseases such as Alzheimer's disease. These are: the uptake of glucose and release of lactate; the uptake of glutamate and release of glutamine; and the uptake of glutathione precursors and release of glutathione. Astrocytes could become promising targets of therapeutic intervention for Alzheimer's disease, if these compromised functions can be normalized with pharmacological agents that are specifically designed to return astrocytes to a quiescent phenotype or to supplement any factors that activated astrocytes fail to produce. PMID- 19903020 TI - CSD, BBB and MMP-9 elevations: animal experiments versus clinical phenomena in migraine. AB - Cortical spreading depression (CSD) has been at the center stage of migraine pathophysiology for approximately six decades. Reanalysis of CSD reveals several major unbridgeable gaps in this experimental neurophysiologic concept for migraine. Key phenotypic and pharmacological features of migraine challenge the assumed pathophysiologic role of CSD. Detection of subclinical infarct-like white matter lesions (WMLs) in the brain of some migraine patients stimulated the concept of CSD-related BBB disruption. Raised plasma levels of matrix metalloproteinases (MMPs) in migraine patients in the headache phase, specifically MMP-9, suggested a pathogenetic role for MMP elevation in the development of both migraine attacks and WMLs. Migraine attacks with or without aura present a unique, profound and protracted vasodilatory challenge to the homeostatic systems of the brain. To accommodate the rather sudden increase in cerebral blood flow, the brain circulatory network must dilate and the BBB must expand considerably. MMPs can influence expansion of the extracellular matrix of the BBB and loosening of the intercellular tight junctions between endothelial cells through proteolytic degradation during migrainous cerebrovascular dilatation. WMLs most probably reflect transient and discrete breakdown of the BBB consequent to sustained cerebral hyperperfusion rather than hypoperfusion. Systemic elevations of MMPs are not specific to migraine but are found in a variety of neurological and extra-neurological disorders. This perspective presents a conceptual dissociation between the effects of CSD on the brain of experimental animals and the clinical phenomena in migraine patients. PMID- 19903021 TI - All-trans retinoic acid as a novel therapeutic strategy for Alzheimer's disease. AB - Retinoic acid, an essential factor derived from vitamin A, has been shown to have a variety of functions including roles as an antioxidant and in cellular differentiation. Since oxidative stress and dedifferentiation of neurons appear to be common pathological elements of a number of neurodegenerative disorders, we speculated that retinoic acid may offer therapeutic promise. In this vein, recent compelling evidence indicates a role of retinoic acid in cognitive activities and anti-amyloidogenic properties. Here, we review the actions of retinoic acid that indicate that it may have therapeutic properties ideally served for the treatment of neurodegenerative diseases such as Alzheimer's disease. PMID- 19903023 TI - MAPK, beta-amyloid and synaptic dysfunction: the role of RAGE. AB - Genetic and biological studies provide strong support for the hypothesis that accumulation of beta amyloid peptide (Abeta) contributes to the etiology of Alzheimer's disease (AD). Growing evidence indicates that oligomeric soluble Abeta plays an important role in the development of synaptic dysfunction and the impairment of cognitive function in AD. The receptor for advanced glycation end products (RAGE), a multiligand receptor in the immunoglobulin superfamily, acts as a cell surface binding site for Abeta and mediates alternations in the phosphorylation state of mitogen-activated protein kinase (MAPKs). Recent results have shown that MAPKs are involved in neurodegenerative processes. In particular, changes in the phosphorylation state of various MAPKs by Abeta lead to synaptic dysfunction and cognitive decline, as well as development of inflammatory responses in AD. The present review summarizes the evidence justifying a novel therapeutic approach focused on inhibition of RAGE signaling in order to arrest or halt the development of neuronal dysfunction in AD. PMID- 19903024 TI - Mediators of tau phosphorylation in the pathogenesis of Alzheimer's disease. AB - The need for disease-modifying drugs for Alzheimer's disease has become increasingly important owing to escalating disease prevalence and the associated socio-economic burden. Until recently, reducing brain amyloid accumulation has been the main therapeutic focus; however, increasing evidence suggests that targeting abnormal tau phosphorylation could be beneficial. Tau is phosphorylated by several protein kinases and this is balanced by dephosphorylation by protein phosphatases. Phosphorylation at specific sites can influence the physiological functions of tau, including its role in binding to and stabilizing the neuronal cytoskeleton. aberrant phosphorylation of tau could render it susceptible to potentially pathogenic alterations, including conformational changes, proteolytic cleavage and aggregation. While strategies that reduce tau phosphorylation in transgenic models of disease have been promising, our understanding of the mechanisms through which tau becomes abnormally phosphorylated in disease is lacking. PMID- 19903022 TI - Can peripheral leukocytes be used as Alzheimer's disease biomarkers? AB - Alzheimer's disease (AD) is the leading cause of dementia in elderly populations throughout the world and its incidence is on the rise. Current clinical diagnosis of AD requires intensive examination that includes neuropsychological testing and costly brain imaging techniques, and a definitive diagnosis can only be made upon postmortem neuropathological examination. Additionally, antemortem clinical AD diagnosis is typically administered following onset of cognitive and behavioral symptoms. As these symptoms emerge relatively late in disease progression, therapeutic intervention occurs after significant neurodegeneration, thereby limiting efficacy. The identification of noninvasive diagnostic biomarkers of AD is becoming increasingly important to make diagnosis more widely available to clinics with limited access to neuropsychological testing or state-of-the-art brain imaging, reduce the cost of clinical diagnosis, provide a biological measure to track the course of therapeutic intervention, and most importantly, allow for earlier diagnosis--possibly even during the prodromal phase--with hopes of therapeutic intervention prior to appreciable neurodegeneration. Circulating leukocytes are attractive candidate AD biomarkers as they can be obtained in a minimally invasive manner and are easily analyzed by widely available flow cytometry techniques. In this review, we critically analyze the potential utility of peripheral leukocytes as biological markers for AD. PMID- 19903025 TI - What is the association between depression and Alzheimer's disease? AB - Depression and dementia, in particular Alzheimer's disease (AD), are critically important issues in the mental health of old age. Both conditions apparently reduce quality of life and increase the impairment of activities of daily living for elderly persons. AD usually shows poor prognosis owing to progressive neuronal degeneration, while depression is basically reversible. However, depressive symptoms are common in AD and occur in approximately 20-30% of patients with AD. Epidemiological studies have shown a possible pathological association between depression and AD. Some longitudinal studies have reported that depression is a prodromal sign or might be both a prodromal symptom of AD and a risk factor. Other studies have suggested that depressive symptoms appear to coincide with or follow the onset of AD rather than precede it. However, it still remains controversial whether depressive symptoms represent a risk factor for AD, whether they are an early symptom of neurodegeneration, or whether they are a reaction to early cognitive deficits. A better understanding of the link between AD and depression might have important clinical and research implications. This review provides an overview of current knowledge regarding a relation between depression and AD and also proposes a research and clinical perspective on depression in AD. PMID- 19903026 TI - Regional anesthesia for acute traumatic injuries in the emergency room. AB - Since the introduction of cocaine in 1884, regional nerve block procedures have been used in anesthesia practice for over 100 years. While almost all medical specialties use simple regional anesthesia techniques, anesthesia providers use a wider variety of more specific nerve block techniques than any other speciality. Anesthesiologists have assumed a vital role in recent military conflicts and, together with surgeons and emergency physicians, have introduced regional anesthesia techniques for the treatment and transport of injured soldiers. While such techniques have only been applied to a limited extent in civilian emergency settings, it is likely that current military experience will enhance future use of regional anesthesia techniques for the care of trauma patients in the civilian prehospital and emergency room settings. PMID- 19903029 TI - Sex, fat and breast cancer. PMID- 19903030 TI - Impact of first trimester and postpartum period thyroid autoantibodies on abortus incidence in Turkish pregnant women. AB - OBJECTIVE: Abortus is a common problem and is observed approximately in one third of all pregnancies. In this study, we aimed to evaluate in euthyroid pregnant population the effect of thyroid autoantibodies of both first trimester and postpartum period on abortus incidence. DESIGN AND METHOD: Euthyroid 128 pregnant women were included in the study. All pregnants were required having a history of maximum one abortus without any previous endocrinological, immunological disorders. Thyroid autoimmunity was defined as having serum levels of either antithyroidperoxidase or antithyroglobulin antibodies higher than 34 and 115 IU/ml, respectively. RESULTS AND CONCLUSIONS: One hundred of 128 pregnant women (78.1%) had no thyroid autoantibodies, whereas 28 pregnant women (21.9%) had positivity for one of thyroid autoantibodies. One hundred pregnant women (78.1%) gave healthy births, whereas 28 (21.9%) had abortus. Abortus incidence was 28.6% in autoantibody positive group, whereas it was 20% in autoantibody negative group. Serum anti-Tg levels both in first trimester and both in postpartum period were higher in pregnants abortus group when compared with healthy births group. Although, we found no correlation between abortus incidence and thyroid autoantibodies presence, first trimester serum levels of anti-Tg levels may have correlated with abortus incidence and may be an indicator of a closer follow-up. In sight of this study, it can be concluded that autoimmunity against thyroid may continue after termination of pregnancy and this was the first study evaluating the postpartum antibodies levels which may be helpful in subsequent pregnancies. PMID- 19903031 TI - Involvement of HSP10 during the ovarian follicular development of polycystic ovary syndrome: Study in both human ovaries and cultured mouse follicles. AB - OBJECTIVE: To study the possible roles of HSP10 in the follicular development. METHODS: In this study, we examined the expression of HSP10 during the follicular development in human ovaries and cultured mouse follicles as well as its functional relevance with PCOS. Ovary tissues from normal adults (n = 3) were obtained with consents. Mouse early antral follicles (diameter: 220-250 mum) were cultured for 3 days in vitro. Western Blot and immunohistochemistry were used to determine the HSP10 expression and localisation during follicular development in vivo and in vitro. RESULTS: HSP10 protein was detected only in oocytes from human preantral follicles, whereas in antral follicles, it was localised in oocytes, granulosa cells, theca cells and stroma cells. Furthermore, in cultured mouse antral follicles, a similar trend of HSP10 expression during follicle development was observed. CONCLUSION: HSP10 expression was increased as larger area and higher level of density during follicular development both in human and mouse follicles cultured in vitro. Our previous studies showed that HSP10 was highly expressed in normal ovaries compared with those from PCOS. The mouse early antral follicle culture approach may help to understand the role of HSP10 in pathophysiological development of PCOS. PMID- 19903032 TI - Comparison of corticoid substitution versus combined oral contraception administration in the treatment of non-classic adrenal hyperplasia: a prospective study. AB - OBJECTIVE: The clinical symptoms of non-classic adrenal hyperplasia (NCAH) are the same as those in patients with polycystic ovary syndrome (PCOS). The aim of our study was to compare conventional corticoid treatment of NCAH with the effect of combined oral contraception (COC) administration (used in treatment of PCOS) on clinical and laboratory parameters of NCAH. DESIGN: A prospective clinical study, cross-over design. MATERIAL AND METHODS: Since 1999 from 298 hyperandrogenic women, eight patients having 21-hydroxylase deficient NCAH have been identified. They were divided equally into two groups according to the order of application treatment modality (hydrocortison vs. COC). Effect of treatment of both modalities on clinical symptoms (hirsutism - FG score, acne, menstrual cycle) and laboratory parameters (testosterone, androstenedione, dehydroepiandrosterone, dehydroepiandrosterone sulphate, sex hormone binding globulin (SHBG)) were evaluated. RESULTS: We observed the decrease of plasma androgens in both groups, which did not differ significantly. Significant increase of SHBG (i.e. decrease of free androgens) was, however, documented in each period with COC administration. Not surprisingly, improvement of the most frequent clinical symptom of NCAH in our study group, oligomenorrhea, was also more apparent in COC. Hirsutism was only a minor problem in our group that did not allow to evaluate treatment effect of both the modalities CONCLUSION: Our results indicate that ovarian suppression by COC administration can effectively suppress androgen production and improve the most frequent clinical symptom (irregular cycle) in patients with NCAH, so can be successfully used for the treatment at least under basal conditions. Whether corticosteroid substitution can be limited to patients with inadequate response to COC on plasma androgen levels or with signs of adrenal insufficiency requires further data. PMID- 19903033 TI - Repressive effect of the phytoestrogen genistein on estradiol-induced uterine leiomyoma cell proliferation. AB - OBJECTIVE: Uterine leiomyomas are the most common gynecological benign tumor and greatly affect reproductive health and well-being. They are the predominant indication for hysterectomy in premenopausal women. Current epidemiological study reported that soy products intake is inversely associated with diseases leading to hysterectomy. Genistein is a soy-derived phytoestrogen and its inhibitory effect on leiomyoma cell proliferation is reported. In this study, we investigated the siginificant inhibitory effect of genistein on estradiol (E(2)) induced leiomyoma cells proliferation. STUDY DESIGN: The Eker rat-derived uterine leiomyoma cell line ELT-3 cells were used. Cell proliferation was assessed by counting the number of cells. The expression of estrogen receptors and peroxisome proliferator-activated receptor-gamma (PPARgamma) was evaluated by Western blot analysis. RESULTS: PPARgamma was expressed in ELT-3 cells and genistein acted as PPARgamma ligand. This inhibitory effect of genistein was attenuated by the treatment of cells with PPARgamma antagonist bisphenol A diglycidyl ether (BADGE) or GW9662. CONCLUSION: These experimental findings in vitro show that the repressive effect of genistein on E(2)-induced ELT-3 cell proliferation is through the activation of PPARgamma. Genistein may be useful as an alternative therapy for leiomyoma. PMID- 19903034 TI - Body mass index is a major determinant of abdominal fat accumulation in pre-, peri- and post-menopausal women. AB - OBJECTIVE: To investigate the role of menopause, body mass index (BMI) and aging on body fat distribution in women. DESIGN: In this population-based cross sectional study, 335 women (126 in pre-menopause, 75 in peri-menopause and 134 in post-menopause according to Stages of Reproductive Aging Workshop criteria) were evaluated for body mass composition and fat distribution by dual X-ray absorptiometry procedure. A sub-group of 79 women with similar age and BMI was extracted from the sample to examine the relative influence of BMI in body fat distribution. RESULTS: ANCOVA analysis of total sample showed an age-independent increase of total fat mass (p < 0.001) and percentage on total weight (p < 0.001), arms fat mass (p < 0.01), legs fat mass percentage on total fat (p < 0.05) and trunk fat mass (p < 0.001) and percentage (p < 0.05) in peri- and post- with respect to pre-menopausal women. In the sub-sample including age and BMI matched women the difference of regional fat parameters among menopausal status was no more statistically significant. CONCLUSION: BMI, and not age, is the main determinant of the increase of body fat mass (total and abdominal) observed during the menopausal transition. PMID- 19903035 TI - The renaissance of metformin in endocrine clinical practice. PMID- 19903036 TI - Juvenile ovarian granulosa cell tumor: a benign or malignant condition? PMID- 19903037 TI - What causes hot flushes? The neuroendocrine origin of vasomotor symptoms in the menopause. AB - Vasomotor symptoms (VMS) such as hot flushes and night sweats are frequently encountered during menopause and can greatly reduce the quality of life. These symptoms are causally related to decreasing estradiol concentrations, mainly in the serum and subsequently also in the hypothalamic temperature regulating centre. The lack of estrogens alters neurotransmitter activity, especially in the serotonergic and noradrenergic pathways. Because sex steroids act as potent neuromodulators, the substitution of ovarian sex steroids by hormone replacement therapy is the most effective treatment option for VMS. When contraindications exist for the use of sex steroids, steroid-free drugs are a possible alternative. A better understanding of the physiology of thermoregulation, thermoregulatory dysfunction and adaptive processes of the brain may facilitate the development of new therapeutic approaches. Such drugs could then be used to treat vasomotor disorders even when the use of steroid hormones is contraindicated. This review article summarises our knowledge on the mechanisms of temperature regulation and describes deviations from this regulation during altered sex steroid conditions. Our current knowledge on neuroendocrinology of thermoregulation may serve as a basis for the use of steroid-free pharmacological intervention. PMID- 19903038 TI - The bone mass density in postmenopausal women using hormonal replacement therapy in relation to polymorphism in vitamin D receptor and estrogen receptor genes. AB - The aims of the study were as follows: (1) To identify the differences in spinal body mass density (BMD) in relation to polymorphism in vitamin D receptor (VDR) and estrogen receptor-alpha (ERalpha) genes in untreated women with postmenopausal osteoporosis. (2) To assess the efficacy of treatment in women with postmenopausal osteoporosis in relation to polymorphism in VDR and ERalpha genes. (3) To find the estradiol concentration necessary to protect bone tissue in patients with a given polymorphism in VDR and ERalpha genes. METHODS: The study included 44 postmenopausal women with primary osteoporosis who used cyclic hormonal replacement therapy (HRT) for a year. The polymorphism of ERalpha and VDR genes were evaluated. We also determined the age, body mass index and spinal BMD before and after 12 months of administration the HRT. RESULTS: We found a significant spinal BMD increase, what is connected with ERalpha genotype and both VDR and ERalpha genes. There is no such a correlation observed in polymorphism of VDR gene. CONCLUSIONS: (1) There is no relationship between VDR and ERalpha genes polymorphism and the stage of osteoporosis related to the spinal BMD value before treatment. (2) The XX, PP or Bb markers or only X, P, B alleles are connected with a significant decrease of treatment efficacy. (3) Estradiol serum concentration before and during HRT is not dependent on the polymorphism of VDR and ERalpha genes. PMID- 19903039 TI - New onset alopecia and hirsutism in a postmenopausal women. AB - Recent onset of hirsutism in postmenopausal women is mostly caused by androgen secretion from adrenal or ovarian tumours. Ovarian hyperthecosis (OH) is a cause of hyperandrogenism in premenopausal women, few cases of postmenopausal presentation have been described. We report on a 73-year old women with androgenic alopecia and hirsutism of recent onset because of elevated testosterone levels. Radiologic imaging showed no tumours of the adrenal glands and ovaries. Careful re-evaluation revealed increased ovarian size in relation to age. Bilateral ovarectomy confirmed the diagnosis of ovarian hyerthecosis and led to improvement of clinical findings. It is important to review imaging findings as OH may elude imaging studies. OH should be included in the differential diagnosis of postmenopausal hyperadnrogenism particularly if androgen excess is of recent-onset. PMID- 19903040 TI - Hyperandrogenemia influences the prevalence of the metabolic syndrome abnormalities in adolescents with the polycystic ovary syndrome. AB - OBJECTIVE: The prevalence of the metabolic syndrome (MBS) abnormalities in Italian adolescents with polycystic ovary syndrome (PCOS) was evaluated. DESIGN: Retrospective chart review. SETTING: University outpatient clinic. PARTICIPANTS: Fifty-three adolescents with PCOS. INTERVENTIONS: Subjects underwent a physical evaluation. Fasting blood samples were taken for the evaluation of metabolic parameters. MAIN OUTCOME MEASURES: The prevalence of MBS abnormalities according to de Ferranti criteria was assessed. RESULTS: 9.4% of adolescents with PCOS had the MBS (three abnormalities). Twelve girls (22.7%) had two abnormalities. Seventeen (32.1%) of PCOS girls have no MBS abnormalities. PCOS adolescents with the MBS were more obese, insulin resistant and they had significantly higher levels of total and free testosterone. The number of metabolic abnormalities correlated with free, total testosterone, free androgen index (FAI) and body mass index (BMI). Groups with two or three abnormalities were not differentiated by BMI, insulin, lipids, blood pressure, but they were differentiated by total and free testosterone and FAI. Adolescents with the MBS have higher total and free testosterone and FAI than girls with two MBS abnormalities. CONCLUSIONS: The MBS and its components are present in some adolescents with PCOS, placing them at increased risk for cardiovascular disease early in adulthood. Hyperandrogenemia is a risk factor for MBS independent of obesity. PMID- 19903041 TI - Sheehan's syndrome and its impact on bone mineral density. AB - INTRODUCTION: Although there have been few studies investigating osteoporosis in isolated hormone deficiencies or other causes of hypopituitarism, the relationship between Sheehan's syndrome (SS) and osteoporosis has not been investigated. In the present study, we aimed to evaluate bone mineral density (BMD) in patients with SS in comparison with healthy women. METHODS: Sixty-one patients with SS and 62 matched healthy controls were included. Biochemical, hormonal assessments and BMD evaluations were carried out in patients and controls, and a subgroup analysis according to menopausal status was done (premenopausal < 50 years; postmenopausal > 50 years). RESULTS: The mean levels of serum anterior pituitary hormones were significantly lower in pre- and postmenopausal patients with SS compared with respective control groups (p < 0.0001). For both pre- and postmenopausal subjects, compared with respective controls, serum calcium and ALP levels, femur-T score, femur-Z score, spine (L1 L5)-T score, spine (L1-L5)-Z score and BMD values were lower, and phosphorus and parathyroid hormone (PTH) levels were higher in patients with SS. CONCLUSIONS: Patients with SS had low BMD. The possible mechanism responsible for osteoporosis may be hypogonadism, growth hormone deficiency and disorders of parathyroid hormone and calcium metabolism. But the contribution of each anterior pituitary hormone deficiency on bone loss should be clarified in further prospective studies. PMID- 19903043 TI - Hepatotoxicity during low-dose flutamide treatment for hirsutism. AB - BACKGROUND: The nonsteroidal antiandrogenic drug flutamide is a safe and generally well-tolerated drug used for the treatment of prostate cancer and female hyperandrogenism. CASE: We describe the case of a 26-year-old girl with amenorrhea and severe hirsutism who was treated with flutamide 250 mg/day and developed liver toxicity during therapy. RESULTS: Other causes of liver toxicity were appropriately ruled out. The use of the standard Council for International Organization of Medical Sciences (CIOMS) scale and the Maria & Vitorino (M&V) scale indicated a highly probable relationship between the development of liver toxicity and flutamide therapy. Severe liver dysfunction has been rarely documented in women with hirsutism treated with flutamide, even though cases of fulminant liver failure have been described. The mechanisms responsible for the occurrence of hepatotoxicity during treatment with flutamide are unknown, but mitochondrial dysfunction seems to be implicated. CONCLUSION: The potential of flutamide to act as a potent hepatotoxin should be kept in mind when treatment with this drug is being planned. This case reminds us that patients who are receiving flutamide should be regularly monitored for liver function. If drug induced liver injury is suspected, flutamide must be discontinued promptly to avoid progression of liver injury. PMID- 19903044 TI - Menopausal hot flush: is it only a nuisance or also a marker of cardiovascular disease risk? AB - Hot flush is one of the most commonly reported symptoms during menopause; however, it is not experienced by all menopausal women, for reasons that remain unclear. In this review, we present current evidence that link hot flushes with cardiovascular disease, suggesting that the persistence of hot flushes many years after the menopause may represent a marker of an underlying disorder that increases the risk for cardiovascular disease. PMID- 19903045 TI - Protection from high-fat-diet-induced impaired glucose tolerance in female Sprague-Dawley rats. AB - In this study, we examined the long-term influence of a high-fat (HF) diet compared with a standard chow (SC) diet on glucose metabolism in genetically normal Sprague-Dawley rats. As the diet was given for a period of 14 months it represents almost a life-long exposure of the rats to the diet. Results showed that (1) after exposure to 14 months of SC or HF feeding, the fasting glucose levels were below 5.1 mmol/l in both genders; (2) the 2 h plasma glucose concentration (2 h PG) of male rats after an intraperitoneal glucose tolerance test for the SC groups and for the HF groups was significantly higher than for females; (3) the liver triglycerides content was significantly enhanced by HF feeding as compared with SC-feeding, and histological staining showed that both genders of the HF group developed severe liver steatosis; (4) the fasting insulin concentration in blood of HF-fed males was significantly higher than that of females and homeostasis model assessment of insulin resistance was also significantly higher; (5) the insulin secretion capabilities in response to glucose stimulation were significantly higher in HF-fed female rats than that of males. Taken together, these data indicate that glucose intolerance produced by a HF diet in rats shows a gender difference, and females are protected against development of impaired glucose tolerance under HF condition. PMID- 19903046 TI - Endometriosis--a European perspective. PMID- 19903047 TI - Endometriosis in Hungary. AB - Endometriosis is a growing healthcare problem all around the world. I discuss in this article how the hungarian healthcare system is working and helping patients suffering from endometriosis. I discuss all diagnostic and therapeutic possibilities which are available for the patients. I discuss also some data from my own practice, 181 patients suffering from chronic pelvic pain caused by endometriosis. PMID- 19903049 TI - Anti muellerian hormone serum levels in women with endometriosis: a case-control study. AB - OBJECTIVE: To compare the anti muellerian hormone (AMH) serum levels in women with and without endometriosis. DESIGN: A case-control study SETTING: Women's General Hospital, Linz, Austria. PATIENT(S): Our study included a total of 909 patients undergoing in vitro fertilisation/intracytoplasmic sperm injection (IVF/ICSI) treatment or consulting our specific endometriosis unit. After proofing the exclusion criteria, 153 of these patients with endometriosis (study group) were matched with 306 patients undergoing IVF/ICSI treatment because of a male factor (control group). INTERVENTIONS: None. MAIN OUTCOME MEASURES: AMH serum level. RESULTS: Mean AMH serum level was significantly lower in the study than in the control group (2.75 + or - 2.0 ng/ml vs. 3.46 + or - 2.30 ng/ml, p < 0.001). In women with mild endometriosis (rAFS I-II), the mean AMH level was almost equal to the control group (3.28 + or - 1.93 ng/ml vs. 3.44 + or - 2.06 ng/ml; p = 0.61). A significant difference in mean AMH serum level was found between women with severe endometriosis (rAFS III-IV) and the control group (2.38 + or - 1.83 ng/ml vs. 3.58 + or - 2.46 ng/ml; p < 0.0001). CONCLUSION: Lower AMH serum levels and an association with the severity were found in women with endometriosis. Physicians have to be aware of this fact. Because of the expected lower response on a controlled ovarian hyperstimulation (COH), AMH serum level should be measured to optimise the dose of gonadotropin treatment previous to a COH, especially in women with severe endometriosis. PMID- 19903048 TI - Correlation between symptoms of pain and peritoneal fluid inflammatory cytokine concentrations in endometriosis. AB - Endometriosis affects 10-20% of women during reproductive age and is a common cause of infertility and pain leading to work absenteeism and reduced quality of life.The objective of this study was to investigate the association between the presence and concentration of interleukin-8 (IL-8), RANTES, osteoprotegerin (OPG), pregnancy-associated plasma protein A (PAPP-A), tumour necrosis factor alpha (TNF-alpha), midkine and glycodelin in the peritoneal fluid (PF) and the intensity of pain reported by patients undergoing laparoscopy in our clinic. They rated their pain during menstruation, intercourse and lower abdominal using a visual analogue scale. During laparoscopy, PF was aspirated. Pain scores were correlated to the concentration of the above substances in the PF and to the stage of endometriosis. Endometriosis was histologically confirmed in 41 of 68 participating women; 27 without such evidence were considered as controls. TNF alpha and glycodelin correlated positively with the level of menstrual pain. For IL-8, RANTES, OPG and PAPP-A no correlation between their PF concentration and the menstrual pain scores was observed. Patients with severe dysmenorrhoea had increased PF cytokine and marker levels; the difference was significant for TNF alpha and glycodelin when compared with the other patients (no or moderate pain). TNF-alpha and glycodelin may thus play a role in endometriosis and the severity of menstrual pain. PMID- 19903050 TI - Impact of endometriosis on quality of life: a pilot study. AB - Endometriosis affects 6-10% of women in reproductive age, 35-50% of whom experience pain, infertility or both. Mild cases are managed medically but surgery provides relief to women in pain. However, symptoms recur in 75% of cases within 2 years. We investigated the impact of endometriosis on quality of life among 65 women aged 18-60 years working at a city supermarket in Giessen, Germany. Of the 65 women, 12 had undergone surgeries, 22 had dysmenorrhoea, 24 dyspareunia and 3 were infertile. Of the 22 women with dysmenorrhoea, 10 had difficulties performing gardening, housework, sports and leisure activities. Five of these 10 women experienced social isolation, 6 professional setbacks; 6 declined efficiency at work and 3 had taken time off work. Of the 24 women with dyspareunia, 7 experienced minimal, 12 light and 5 moderate to strong pain. Only 16 of these 24 women discussed the problem with their partners. This study demonstrates that pain is a major cause of physical, psycho-social, emotional and professional or work related impairment among women with endometriosis. Because endometriosis is likely to impose emotional and financial burdens, we suggest that future studies should be extended to include interviews with family members. PMID- 19903052 TI - Expression of the human endogenous retroviruse-W envelope gene syncytin in endometriosis lesions. AB - BACKGROUND: None of the existing theories provides a satisfactory explanation of the development of endometriosis. One hypothesis that may lead to further clarification is that the expression of specific proteins of human endogenous retroviruses (HERVs) might influence the development of endometriosis lesions. Such endogenous retroviral proteins include syncytin, coded by HERV-W, which is associated with the physiological development of the placenta during pregnancy. This study investigated the influence of HERV-W gene expression in endometriosis foci (EM) quantitatively at the RNA level. MATERIALS AND METHODS: Specific RNA expression of syncytin (HERV-W) was investigated in various endometrial tissues from 42 patients (with normal endometrium, postmenopausal endometrium, EM, and endometrial carcinoma). RNA was isolated from the tissue samples and transcribed into DNA using reverse transcriptase polymerase chain reaction. The resulting DNA fragments were analyzed using agarose gel electrophoresis and assessed quantitatively. RESULTS: Normalized syncytin expression was low in EM. In Histologically normal endometrium from endometriosis patients, the expression of normalized syncytin was seven times higher in comparison with the histologically normal endometrium in the control group. CONCLUSIONS: HERV-W syncytin expression apparently does not play a role in EM. However, it may possibly influence the development of endometriosis because of increased expression in normal endometrium in endometriosis patients. PMID- 19903051 TI - Neurokinin 1 receptor gene polymorphism might be correlated with recurrence rates in endometriosis. AB - INTRODUCTION: Dysmenorrhoea is the major symptom in women with endometriosis. Recently, pain modulation through Neurokinin-1-receptor (NK1R) pathways have been investigated in neuropathic pain patients. Aim of this study was, therefore, to examine the effect of a single nucleotide polymorphism (SNP) of the NK1R gene on the susceptibility for endometriosis and the disease free survival (DFS) after surgery for endometriosis. MATERIAL AND METHODS: A case-control study was conducted and germline DNA was isolated. Patients were followed up for a recurrence of the disease up to 4 years. Case-control analyses were performed for parameters of the medical history and the genotype of the NK1R-SNP rs881. Furthermore, DFS probabilities were calculated. RESULTS: Concerning the DFS preoperative pain levels and the NK1R genotype were independent predictors for a recurrence with hazard ratios of 2.55 (95% CI: 1.32-4.95) for patients with a high preoperative pain level and 0.44 for patients with a heterozygous or homozygous variant genotype in rs881 (95% CI: 0.21-0.88). CONCLUSION: The polymorphism rs811 seems to be associated with a lower recurrence risk in endometriosis patients. Thus, there might be a clinical relevant role of the NK1 pathway in the pain perception of endometriosis patients. PMID- 19903053 TI - Endometriosis in the north Italian province of South Tyrol. AB - Here, we report regarding the health status and medical support for patients suffering from endometrioisis in South Tyrol, which is a politically autonomous province in the north of Italy containing three different ethnic groups. The health service is administered largely by the autonomous regional government. Because of the establishment of a centre for reproductive medicine and the introduction of laparoscopy as a prime surgery method, the gynaecological department of the hospital in Bruneck developed into a reference centre for diagnosis and surgical treatment for endometriosis. The planned future social, health care and insurance developments on local and national level regarding this illness will be discussed. PMID- 19903054 TI - Progestins and medical treatment of endometriosis - physiology, history and society. AB - The transitory effect of hormonal treatment is the alleged main reason to criticize progestins (PGS) and combined pills (OP) in the managment of endometriosis. To the contrary their poor efficacy in the long run is often underlined. As a result, medical treatment is too seldom advised in endometriosis. In this article, we shall focus on the analysis of the reasons of the paucity of the medical interest given to progestins, reasons, which are not of a scientific or objective nature. The ultimate aim of this analysis is to develop arguments in favour of continuous administration of hormones as to obtain not simply an anovulation but a state of prolonged amenorrhea much more efficacious than the simple suppression of ovulation too often advised. And, with an emphasis on the fundamental role of surgery in the treatment of endometriosis, to give the greatest consideration to the specific nature of this disease, which is a chronic disease, justifying the long duration of hormonal administration. PMID- 19903055 TI - Minimal endometriosis: a therapeutic dilemma? AB - Accidental finding of minimal endometriosis during surgery for complaints unassociated with endometriosis presents a therapeutical dilemma. Because the clinical significance of minimal endometriosis is not thoroughly defined, it is uncertain by which means, if at all, such types of peritoneal or ovarian lesions should be treated. We, therefore, compiled our clinical observations and evaluated them in relation to the results reported in the recent literature. A search on the clinical importance and the need for any treatment of minimal and mild endometriosis was conducted. Based on the available evidence, we are tempted to conclude that minimal endometriosis should be treated surgically when accidentally discovered. This approach should be exerted even in the absence of clinical symptoms. However, postoperative medical treatment is not warranted in those patients who are without clinical complaints. Because the clinical course of minimal endometriosis is not predictable, any benefit from specific medications remains uncertain. When associated with infertility, minimal endometriosis should be surgically erased, thus to allow spontaneous conceptions to occur. As there is no evidence of medical treatment modalities altering the clinical course of minimal and mild endometriosis, any specific medical treatment (i.e. GnRH analogues, danazol) is not indicated in asymptomatic patients and those desiring pregnancy. PMID- 19903056 TI - Quality of life impairment among postmenopausal women varies according to race. AB - BACKGROUND: Few studies have addressed the impact of menopausal symptom severity over quality of life (QoL) in Latin American women with different ethnics. OBJECTIVE: To assess menopausal symptom severity and the QoL among postmenopausal Colombian women with three different ethnicities. METHOD: Data of healthy naturally occurring postmenopausal Hispanic, indigenous and black women aged 40 59 years who participated in a cross-sectional study filling out the Menopause Rating Scale (MRS) and a general questionnaire was analysed. RESULTS: A total of 579 women were included, 153 Hispanic, 295 indigenous and 131 Afro-descendent. Hispanic women had an average age of 55.3 +/- 3.3 years. Indigenous and black women were less educated than the Hispanic ones (2.2 +/- 1.8 and 4.6 +/- 4.4 vs. 6.4 +/- 3.5 years, p < 0.0001). Hispanic women displayed lower total MRS scores (better QoL) when compared to indigenous and black women. Urogenital scoring was worse among indigenous women compared to Hispanic and black women. Black women presented higher MRS psychological and somatic scorings than Hispanic and indigenous women. After adjusting for confounding factors, indigenous and black women continued to display a higher risk for impaired QoL, total MRS score > 16 (OR: 3.11, 95% CI: 1.30-7.44 and OR: 5.29, 95% CI: 2.52-11.10, respectively), which was significantly higher among indigenous women due to urogenital symptoms (OR: 102.75, 95% CI: 38.33-275.47) and black women due to psychological (OR: 6.58, 95% CI: 3.27-13.27) and somatic symptoms (OR: 3.88, 95% CI: 1.83-8.22). CONCLUSION: In this postmenopausal Colombian series, menopausal symptoms in indigenous (urogenital) and black (somatic/psychological) women were more severe (impaired QoL) when compared to Hispanic ones. PMID- 19903057 TI - Treatment of osteoporosis by long-term magnetic field with extremely low frequency in rats. AB - BACKGROUND: Osteoporosis is characterised by low bone mass and structural deterioration of bone tissue. In this study, the role of long-term extremely low frequency magnetic field (ELFMF) on osteoporosis was evaluated. METHODS: The experiments were performed on 45 female Sprague-Dawley rats. The rats were divided into three groups (n = 15): Group I (ovariectomy (OVX) + ELFMF exposure), Group II (ovariectomised rats did not receive any treatment) and Group III (cage control). Six months, 50 Hz, 1.5 mT magnetic field (MF) was used on Group I and Group II. Total body images of the animals were obtained with dual-energy X-ray absorptiometry. RESULTS: Bone mineral content (BMC) and bone mineral density values were increased significantly in ELFMF group, decreased in the group of OVX and not changed in cage-control. At the end of the 6 months after exposure with ELFMF, alteration in studied biochemical markers were detected significant. Bone specific alkaline phosphatase (BAP) levels were increased in ELFMF and decreased in OVX groups when compared with cage-control group. N-telopeptide levels in OVX group were significantly higher than other groups. Testosterone and cortisol levels in OVX group were significantly higher and estradiol was lower than other groups. CONCLUSIONS: Our study suggests that ELFMF may be useful in the prevention and treatment of osteoporosis. PMID- 19903058 TI - Oral stomatitis induced by endogenous progesterone: Case report. AB - Oral stomatitis induced by endogenous progesterone is a rare clinical condition which may be associated with cutaneous involvement. That is probably due to the peak of progesterone production during the luteal phase of the menstrual cycle. In the present case report, a 21-year-old patient displayed recurrent ulcerative lesions located on the buccal mucosa or the upper lip, on a monthly basis since the age of 15. Such lesions would always manifest themselves on the second day until the end of the menstrual cycle. PMID- 19903059 TI - The 'RNA-binding ome': future implications for chemotherapeutic efficacy. PMID- 19903060 TI - Stereotactic body radiation therapy for recurrent head & neck cancers: rethinking nonoperative salvage strategies. PMID- 19903061 TI - Targeting of AML-leukemic stem cells with monoclonal antibodies. PMID- 19903063 TI - Pazopanib: an antiangiogenic drug in perspective. AB - Pazopanib, a tyrosine kinase inhibitor targeted to angiogenesis, has been tested in preclinical and clinical trials and has shown promising activity against a variety of solid tumors, such as renal cancer, all of which are related to the angiogenic pathway. It has a safety profile related to this mechanism of action. Diarrhea, hypertension, hair depigmentation and nausea are the most common side effects. Pazopanib is currently under evaluation as monotherapy and in combination with some potentially synergistic agents of proven activity. PMID- 19903064 TI - Nimotuzumab in pediatric glioma. AB - High-grade gliomas and diffuse brainstem gliomas carry a very poor prognosis despite current therapies, and account together for the largest number of deaths in children with brain tumors. Many of these tumors have been found to overexpress the EGF receptor (EGFR). Nimotuzumab (h-R3) is a humanized monoclonal antibody against the EGFR, and consequently inhibits tyrosine kinase activation. In vitro and in vivo studies have supported the antiproliferative, antiangiogenic, pro-apoptotic and radiosensitizing activities of nimotuzumab. Emerging trials suggest a promising role for nimotuzumab as a therapeutic agent in patients with high-grade gliomas. This review attempts to provide a context for the evolving interest and evidence for nimotuzumab in pediatric glioma. PMID- 19903065 TI - Role of prolonged mitotic checkpoint activation in the formation and treatment of cancer. AB - Mitotic abnormalities are a common feature of human cancer cells, and recent studies have provided evidence that such abnormalities may play a causative, rather than merely incidental role, in tumorigenesis. One such abnormality is prolonged activation of the mitotic checkpoint, which can be provoked by a number of the gene changes that drive tumor formation. At the same time, antimitotic chemotherapeutics exert their clinical efficacy through the large-scale induction of prolonged mitotic checkpoint activation, indicating that mitotic arrest is influential in both the formation and treatment of human cancer. However, how this influence occurs is not well understood. In this perspective, we will discuss the current evidence in support of the potential mechanisms by which prolonged activation of the mitotic checkpoint affects both tumorigenesis and antimitotic chemotherapy. PMID- 19903066 TI - Uptake of colorectal cancer screening: system, provider and individual factors and strategies to improve participation. AB - Colorectal cancer (CRC) accounts for 9% of all new cancer cases worldwide and affects over 1 million people each year. Screening can reduce the mortality associated with the disease, yet participation rates are suboptimal. Compliers with CRC screening are less deprived; they have higher education than noncompliers and tend to be male, white and married. Likely reasons for nonparticipation encompass several 'modifiable' factors that could be targeted in interventions aimed at increasing participation rates. Successful intervention strategies include organizational changes, such as increasing access to fecal occult blood test (FOBT) kits, providing reminders to healthcare providers or users about screening opportunities, and educational strategies to improve awareness and attitudes towards CRC screening. Multifactor interventions that target more than one level of the screening process are likely to have larger effects. The biggest challenge for future research will be to reduce inequalities related to socio-economic position and ethnicity in the uptake of screening. PMID- 19903069 TI - Challenges in defining predictive markers for response to endocrine therapy in breast cancer. AB - Endocrine therapy is a major treatment modality for hormone-dependent breast cancer. It has a relatively low morbidity, and there is evidence that antihormonal treatments have had a significant effect in reducing mortality for breast cancer. Despite this, resistance to endocrine therapy, either primary or acquired during treatment, occurs in the majority of patients, and is a major obstacle to optimal clinical management. There is therefore an urgent need to identify, on an individual basis, those tumors that are most likely to respond to endocrine therapy (so sparing patients with resistant tumors the needless side effects of ineffective therapy), and the mechanisms of resistance in tumors that are nonresponsive to treatment (so these can be bypassed). These needs are the focus of this review, which discusses the particular issues encountered when investigating the potential of multigene expression signatures as predictive factors for response to aromatase inhibitors, which have recently become front line endocrine therapies for postmenopausal patients with breast cancer. PMID- 19903068 TI - Molecular mechanisms of castration-resistant prostate cancer progression. AB - Hormone-refractory prostate cancer is the result of regrowth of prostate cancer cells that have adapted to the hormone-deprived environment of the prostate. The process by which castration-resistant prostate cancer (CRPC) cells are generated appears to be varied. The complex mechanism of hormone resistance has been the topic of research in most laboratories that have analyzed the process from different angles. This review compiles research findings that explain the methods of development of hormone resistance in prostate cancer. Research data show many different processes to be involved in the acquisition of hormone resistance. Interestingly, one observes interdependence between these processes, indicating a complex network at play in the development of hormone resistance. Cytokines such as IL-6 have been shown to initiate an alternative signaling pathway, compared with the androgen receptor signaling pathway, in CRPC. IL-6 has been proposed to be the effector of the intracrine signaling pathway by influencing the levels of metabolic enzymes. Neuroendocrine cells are present at low levels in normal prostate, and signify the transitory phase of normal hormone-sensitive cells to hormone-refractory cells. IL-6 induces growth of neuroendocrine cells or neuroendocrine-like features in cells in CRPC. The increased presence of neuroendocrine cells in CRPC signifies a change in the prostate cell microenvironment. The stromal microenvironment also influences the development of CRPC in the hormone-refractory stage. In addition, intracrine androgen metabolic enzymes play a significant role in the development of the hormone refractory process. Despite hormone ablation, there is a residual level of hormones in cells due to active intracrine metabolic pathways. It is acknowledged that the androgen receptor plays the most influential role in development of prostate cancer. In addition to mutation and amplification, the androgen receptor has been characterized and shown to differ in sequence in CRPC compared with the androgen sensitive prostate cancer cells. These variants of the androgen receptor through sequence changes may preserve the basic function of the molecule, but have far reaching consequences on the cell as a whole. A multicombinatorial drug treatment approach has been suggested to target these multiple pathways in an effort to reduce the possibility of recurrence of CRPC. PMID- 19903067 TI - Etiologic factors in testicular germ-cell tumors. AB - Globally, testicular cancer incidence is highest among men of northern European ancestry and lowest among men of Asian and African descent. Incidence rates have been increasing around the world for at least 50 years, but mortality rates, at least in developed countries, have been declining. While reasons for the decreases in mortality are related to improvements in therapeutic regimens introduced in the late 1970s, reasons for the increase in incidence are less well understood. However, an accumulating body of evidence suggests that testicular cancer arises in fetal life. Perinatal factors, including exposure to endocrine disrupting chemicals, have been suggested to be related to risk. PMID- 19903071 TI - Fetal cell microchimerism in cancer: a meaningful event? AB - The influence of pregnancy on the occurrence and evolution of maternal tumors has been long debated. Breast carcinomas or melanomas have been suspected to be more severe during gestation. Recently, many investigators have described the transfer and persistence of fetal cells in maternal circulation and tissues during and after pregnancy. These fetal microchimeric cells have been described in a variety of maternal injured tissues where they displayed the host-tissue phenotype. Given the wide variety of injury and tissue types described, cancer has appeared as a potential situation that could be influenced by fetal microchimeric cells. This new unexplored effect of gestation on tumor course has been hypothesized as either protective against cancer, via the activity of allogenic fetal cells, or as promoting cancer, via a supportive role of fetal microchimeric cells in the tumor stroma. In this review, we will detail recent data supporting these hypotheses. PMID- 19903070 TI - Role of CSF-1 in progression of epithelial ovarian cancer. AB - Despite the dismal outcome seen in the majority of epithelial ovarian cancer patients, there is ongoing progress in understanding the disease at a molecular level. Elucidation of pathways underlying disease progression and metastasis of ovarian cancer is key to development of targeted therapeutics. It is only in this way that therapeutic potential can be translated to reality. Here, we describe the evidence to date for the role of CSF-1/c-fms signaling in ovarian cancer invasiveness and metastasis, including the recent understanding of how CSF-1/c fms expression is regulated with identification of significant post transcriptional regulators. PMID- 19903072 TI - Molecular and cellular biology of rhabdomyosarcoma. AB - Rhabdomyosarcoma is a group of soft-tissue sarcomas that share features of skeletal myogenesis, but show extensive heterogeneity in histology, age and site of onset, and prognosis. This review matches recent molecular data with biological features of rhabdomyosarcoma. Alterations in molecular pathways, animal models, cell of origin and potential new therapeutic targets are discussed. PMID- 19903074 TI - Urokinase plasminogen activator system as a potential target for cancer therapy. AB - Proteolysis of extracellular matrix (ECM) and basement membrane is an essential mechanism used by cancer cells for their invasion and metastasis. The ECM proteinases are divided into three groups: metalloproteinases, cysteine proteinases and serine proteinases. The urokinase plasminogen activator (uPA) system is one of the serine proteinase systems involved in ECM degradation. Members of this system, including uPA and its receptor (uPAR), are overexpressed in several malignant tumors. This system plays a major role in adhesion, migration, invasion and metastasis of cancer cells, thus making it an important target for anticancer drug therapy. Several strategies, including the use of antisense oligodeoxynucleotides, ribozymes, DNAzyme, RNAi, uPA inhibitors, soluble uPAR, catalytically inactive uPA fragments, synthetic peptides and synthetic hybrids are under study, as they interfere with the expression and/or activity of uPA or uPAR in tumor cells. Herein, we discuss the various pharmaceutical strategies under investigation to combat the uPA activity in cancer. PMID- 19903073 TI - Targeting phospholipase D with small-molecule inhibitors as a potential therapeutic approach for cancer metastasis. AB - Phospholipase D (PLD)1 and PLD2, the classic mammalian members of the PLD superfamily, have been linked over the past three decades to immune cell function and to cell biological processes required by cancer cells for metastasis. However, owing to the lack of effective small-molecule inhibitors, it has not been possible to validate these roles for the PLDs and to explore the possible utility of acute and chronic PLD inhibition in vivo. The first such inhibitors have recently been described and demonstrated to block neutrophil chemotaxis and invasion by breast cancer cells in culture, increasing the prospects for a new class of therapeutics for autoimmune disorders and several types of metastatic cancer. PMID- 19903078 TI - Multimodality and interactivity: connecting properties of serious games with educational outcomes. AB - Serious games have become an important genre of digital media and are often acclaimed for their potential to enhance deeper learning because of their unique technological properties. Yet the discourse has largely remained at a conceptual level. For an empirical evaluation of educational games, extra effort is needed to separate intertwined and confounding factors in order to manipulate and thus attribute the outcome to one property independent of another. This study represents one of the first attempts to empirically test the educational impact of two important properties of serious games, multimodality and interactivity, through a partial 2 x 3 (interactive, noninteractive by high, moderate, low in multimodality) factorial between-participants follow-up experiment. Results indicate that both multimodality and interactivity contribute to educational outcomes individually. Implications for educational strategies and future research directions are discussed. PMID- 19903083 TI - Lumenal part of the DC-LAMP protein is not required for induction of antigen specific T cell responses by means of antigen-DC-LAMP messenger RNA electroporated dendritic cells. AB - Previous studies showed that stimulation of T cells derived from HIV-1-infected patients with autologous dendritic cells electroporated with mRNA encoding HIV antigens can induce antigen-specific T cell responses in vitro. Linking the antigen to an MHC class II-targeting sequence, such as dendritic cell lysosome associated membrane protein (DC-LAMP), in the mRNA construct results in presentation of antigenic peptides in both MHC class I and class II molecules and therefore enhances the induced T cell responses. To analyze whether the lumenal domain of DC-LAMP is required for optimal induction of cellular immunity against HIV antigens, we compared fusion constructs with or without the lumenal domain of the DC-LAMP protein. A human codon-optimized consensus Gag sequence and a chimeric cDNA sequence encompassing Tat, Rev, and Nef codons (TaReNef ) were cloned into a vector containing the DC-LAMP sequence with or without its lumenal domain. The Gag protein lacking the DC-LAMP-derived sequence altogether elicited only weak T cell responses. DCs electroporated with Gag or TaReNef linked to DC LAMP were able to elicit similar levels of antigen-specific CD4(+) and CD8(+) T cell responses for both Gag and TaReNef, irrespective of the addition of the DC LAMP lumenal domain. These data show that DC-LAMP-mediated antigen targeting is absolutely required for optimal T cell stimulation, but that in our experimental setup, the lumenal part of DC-LAMP does not improve the overall induction of antigen-specific T cell responses. PMID- 19903075 TI - Clinical implications of near-infrared fluorescence imaging in cancer. AB - Near-infrared (NIR) fluorescence cancer imaging is a growing field for both preclinical and clinical application to the clinical management for cancer patients due to its advantageous features, including a high spatial resolution, portability, real-time display and detailed molecular profiling with the multiplexed use of fluorescent probes. In this review, we present a basic concept of NIR fluorescence imaging and overview its potential clinical applications for in vivo cancer imaging, including cancer detection/characterization, lymphatic imaging (sentinel lymph node detection) and surgical/endoscopic guidance. NIR fluorescence imaging can compensate some limitations of conventional imaging modalities, and thus it could play an important role for cancer imaging combined with other modalities in clinical practice. PMID- 19903084 TI - Transcriptomics of traumatic brain injury: gene expression and molecular pathways of different grades of insult in a rat organotypic hippocampal culture model. AB - Traumatic brain injury (TBI) is the one of the most common forms of head trauma, and it remains a leading cause of death and disability. It is known that the initial mechanical axonal injury triggers a complex cascade of neuroinflammatory and metabolic events, the understanding of which is essential for clinical, translational, and pharmacological research. These can occur even in mild TBI, and are associated with several post-concussion manifestations, including transiently heightened vulnerability to a second insult. Recent studies have challenged the tenet that ischemia is the ultimate modality of tissue damage following TBI, as metabolic dysfunction can develop in the presence of normal perfusion and before intracranial hypertension. In order to elucidate the cellular and molecular changes occurring in TBI as a direct result of neuronal injury and in the absence of ischemic damage, we performed a microarray analysis of expressed genes and molecular interaction pathways for different levels of severity of trauma using an in-vitro model. A stretch injury, equivalent to human diffuse axonal injury, was delivered to rat organotypic hippocampal slice cultures, and mRNA levels following a 10% (mild) and 50% (severe) stretch were compared with controls at 24 h. More genes were differentially expressed following 10% stretch than 50% stretch, indicating the early activation of complex cellular mechanisms. The data revealed remarkable differential gene expression following mTBI, even in the absence of cell damage. Pathway analysis revealed that molecular interactions in both levels of injury were similar, with IL-1beta playing a central role. Additional pathways of neurodegeneration involving RhoA (ras homolog gene family, member A) were found in 50% stretch. PMID- 19903085 TI - Functional properties of cell-seeded three-dimensionally woven poly(epsilon caprolactone) scaffolds for cartilage tissue engineering. AB - Articular cartilage possesses complex mechanical properties that provide healthy joints the ability to bear repeated loads and maintain smooth articulating surfaces over an entire lifetime. In this study, we utilized a fiber-reinforced composite scaffold designed to mimic the anisotropic, nonlinear, and viscoelastic biomechanical characteristics of native cartilage as the basis for developing functional tissue-engineered constructs. Three-dimensionally woven poly(epsilon caprolactone) (PCL) scaffolds were encapsulated with a fibrin hydrogel, seeded with human adipose-derived stem cells, and cultured for 28 days in chondrogenic culture conditions. Biomechanical testing showed that PCL-based constructs exhibited baseline compressive and shear properties similar to those of native cartilage and maintained these properties throughout the culture period, while supporting the synthesis of a collagen-rich extracellular matrix. Further, constructs displayed an equilibrium coefficient of friction similar to that of native articular cartilage (mu(eq) approximately 0.1-0.3) over the prescribed culture period. Our findings show that three-dimensionally woven PCL-fibrin composite scaffolds can be produced with cartilage-like mechanical properties, and that these engineered properties can be maintained in culture while seeded stem cells regenerate a new, functional tissue construct. PMID- 19903087 TI - The evaluation of mineralized collagen as a carrier for the osteoinductive material COLLOSS((r))E, in vivo. AB - In this study, the suitability of mineralized collagen, as a carrier for the native bone morphogenetic protein containing product COLLOSS((r))E, was evaluated in a rat subcutaneous implantation model. Tubular-shaped scaffolds were made from mineralized collagen and were left empty (controls), filled with 40 mg COLLOSS((r))E, or filled with a mixture of COLLOSS((r))E and osteoconductive beta tricalcium phosphate (beta-TCP) granules. Six of all the samples were implanted in 18 animals. During the implantation period, macroscopic examination suggested inflammatory reaction. Histological evaluation after 12 weeks confirmed the presence of inflammatory cells. Although the mineralized scaffold and beta-TCP granules could be clearly seen, no new bone formation was found. This is in contrast to previous reports in a similar set-up wherein a titanium mesh was used as scaffold material in combination with COLLOSS((r))E. It is concluded, therefore, that the osteoinductive capacity of bone morphogenetic proteins or, more specifically, COLLOSS((r))E is inhibited when the carrier gives rise to a significant inflammatory reaction. Additional studies are necessary to assess the effect of combining COLLOSS((r))E with a dimensionally stable and osteoconductive material like beta-TCP. PMID- 19903086 TI - New challenges for intervertebral disc treatment using regenerative medicine. AB - The development of tissue engineering therapies for the intervertebral disc is challenging due to ambiguities of disease and pain mechanisms in patients, and lack of consensus on preclinical models for safety and efficacy testing. Although the issues associated with model selection for studying orthopedic diseases or treatments have been discussed often, the multifaceted challenges associated with developing intervertebral disc tissue engineering therapies require special discussion. This review covers topics relevant to the clinical translation of tissue-engineered technologies: (1) the unmet clinical need, (2) appropriate models for safety and efficacy testing, (3) the need for standardized model systems, and (4) the translational pathways leading to a clinical trial. For preclinical evaluation of new therapies, we recommend establishing biologic plausibility of efficacy and safety using models of increasing complexity, starting with cell culture, small animals (rats and rabbits), and then large animals (goat and minipig) that more closely mimic nutritional, biomechanical, and surgical realities of human application. The use of standardized and reproducible experimental procedures and outcome measures is critical for judging relative efficacy. Finally, success will hinge on carefully designed clinical trials with well-defined patient selection criteria, gold-standard controls, and objective outcome metrics to assess performance in the early postoperative period. PMID- 19903089 TI - Novel freeze-drying methods to produce a range of collagen-glycosaminoglycan scaffolds with tailored mean pore sizes. AB - The pore structure of three-dimensional scaffolds used in tissue engineering has been shown to significantly influence cellular activity. As the optimal pore size is dependant on the specifics of the tissue engineering application, the ability to alter the pore size over a wide range is essential for a particular scaffold to be suitable for multiple applications. With this in mind, the aim of this study was to develop methodologies to produce a range of collagen glycosaminoglycan (CG) scaffolds with tailored mean pore sizes. The pore size of CG scaffolds is established during the freeze-drying fabrication process. In this study, freezing temperature was varied (-10 degrees C to -70 degrees C) and an annealing step was introduced to the process to determine their effects on pore size. Annealing is an additional step in the freeze-drying cycle that involves raising the temperature of the frozen suspension to increase the rate of ice crystal growth. The results show that the pore size of the scaffolds decreased as the freezing temperature was reduced. Additionally, the introduction of an annealing step during freeze-drying was found to result in a significant increase (40%) in pore size. Taken together, these results demonstrate that the methodologies developed in this study can be used to produce a range of CG scaffolds with mean pore sizes from 85 to 325 microm. This is a substantial improvement on the range of pore sizes that were possible to produce previously (96-150 microm). The methods developed in this study provide a basis for the investigation of the effects of pore size on both in vitro and in vivo performance and for the determination of the optimal pore structure for specific tissue engineering applications. PMID- 19903088 TI - Effect of matrix elasticity on the maintenance of the chondrogenic phenotype. AB - The aim of this study was to examine the influence of matrix elasticity on the maintenance of the chondrogenic phenotype of chondrocytes cultured in monolayer. We used a two-dimensional culturing system in which polyacrylamide gels with different concentrations of bis-acrylamide were coated with collagen type I. Matrices with a Young's modulus of 4, 10, 40, and 100 kPa were produced, as determined by atomic force microscopy. Porcine chondrocytes were cultivated on these matrices at a low density for 7 days. The proliferation of cells was analyzed by 5-Bromo-2'-deoxy-uridine incorporation. Maintenance of the chondrogenic phenotype was analyzed by measuring collagen type I, type II, and aggrecan gene expression, immunofluorescence staining for collagen type II, and phalloidin staining for actin filaments. Cellular proliferation and actin organization were decreased on matrices of 4 kPa compared with stiffer substrates. The differentiated phenotype of the chondrocytes grown on matrices of 4 kPa was stabilized, indicated by higher collagen type II and aggrecan, and lower collagen type I expression. These findings indicate that chondrocytes sense the elasticity of the matrix and might be used for the design of scaffolds with mechanical properties specifically tailored to support the chondrogenic phenotype in tissue engineering applications. PMID- 19903090 TI - Epithelial-mesenchymal transition: from molecular mechanisms, redox regulation to implications in human health and disease. AB - Epithelial to mesenchymal transition (EMT) is a fundamental process, paradigmatic of the concept of cell plasticity, that leads epithelial cells to lose their polarization and specialized junctional structures, to undergo cytoskeleton reorganization, and to acquire morphological and functional features of mesenchymal-like cells. Although EMT has been originally described in embryonic development, where cell migration and tissue remodeling have a primary role in regulating morphogenesis in multicellular organisms, recent literature has provided evidence suggesting that the EMT process is a more general biological process that is also involved in several pathophysiological conditions, including cancer progression and organ fibrosis. This review offers first a comprehensive introduction to describe major relevant features of EMT, followed by sections dedicated on those signaling mechanisms that are known to regulate or affect the process, including the recently proposed role for oxidative stress and reactive oxygen species (ROS). Current literature data involving EMT in both physiological conditions (i.e., embryogenesis) and major human diseases are then critically analyzed, with a special final focus on the emerging role of hypoxia as a relevant independent condition able to trigger EMT. PMID- 19903091 TI - Bisphosphonate (zoledronic acid)-induced osteonecrosis of the jaw. AB - OBJECTIVE: The use of bisphosphonates (zoledronic acid) in the treatment of metastatic bone disease has been raised during recent years. The purpose of this treatment is mainly to reduce skeletal-related events, e.g. pain and pathological fractures. The aim of this study was to report the incidence of bisphosphonate related osteonecrosis of the jaw (ONJ). MATERIAL AND METHODS: All patients treated with bisphosphonates during a 5-year period (2003-2007) were reviewed. RESULTS: Fifty-three patients, median age 69 years (range 56-81 years) were treated with bisphosphonates during the period. Fifty-one patients had hormone refractory metastatic prostate cancer and two women had metastatic renal cell carcinoma. During this 5-year period, 686 treatments with bisphosphonates were administrated. The average treatment duration was 14 months (range 1-40 months) with administration of 4 mg of bisphosphonates every 4 weeks. Two cases of ONJ were registered. In the first case, the patient developed spontaneous osteonecrosis, whereas the second patient developed symptoms after a dental procedure. Since the initiation of a routine maxillofacial examination before treatment with bisphosphonates, no ONJ has been seen. CONCLUSION: ONJ is a rare but a very serious complication in relation to treatment with bisphosphonates. To decrease the incidence of ONJ, a maxillofacial examination could be performed in all patients before treatment with bisphosphonates. PMID- 19903092 TI - Combinations of urine-based tumour markers in bladder cancer surveillance. AB - OBJECTIVE: The aim of the study was to investigate whether combinations of urine based tumour markers including urinary cytology (Cytology or Cyt) increase the sensitivity in the detection of bladder cancer recurrence. MATERIAL AND METHODS: Urinary cytology, NMP22, UroVysion (FISH) and ImmunoCyt (uCyt+) were determined in 221 patients during the follow-up of non-muscle-invasive transitional cell carcinoma (NMI TCC) before cystoscopy (n = 49) or with the suspicion of TCC recurrence before transurethral resection of the bladder (n = 173). For all markers alone as well as in all possible combinations (multimarker panels, MPs) sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy were evaluated. MPs were considered positive if at least one marker was positive. RESULTS: No malignancy was found in 108 patients, whereas recurrent TCC was confirmed in 113 patients. Sensitivity and specificity for Cytology were 84% and 62%, for NMP22 68% and 49%, for FISH 72% and 63%, and for uCyt+ 73% and 62%, respectively. The NPV was below 80% for all markers alone. Combinations of two and three markers increased the sensitivity as well as the NPV to over 90 and 80%, by reducing specificity to an average of 44% and 35%, respectively. The most sensitive combinations were NMP22, uCyt+ together with Cytology and FISH, and uCyt+ together with NMP22 (sensitivity for both combinations 98%). There was no further improvement when all four markers were combined. CONCLUSIONS: Combinations of tumour markers increased the sensitivity and NPV in the detection of recurrence of NMI TCC. A stepwise approach of tumour marker determination may be used to reduce the frequency of follow-up cystoscopies at a reasonable risk. PMID- 19903093 TI - Perceived difficulties using everyday technology after acquired brain injury: influence on activity and participation. AB - Using everyday technology (ET) is a prerequisite for activities and participation at home and in the community. It is well known that persons with an acquired brain injury (ABI) can have limitations in activities of daily living but our knowledge of their difficulties using ET is not known. Thirty-six persons (27 men and 9 women, mean age 44 years, age range 26-60) with an ABI (2-10 years post injury) were interviewed, using the Everyday Technology Use Questionnaire (ETUQ), about their perceived difficulties using ET and how these difficulties influenced their everyday activities and their possibilities to participate at home and in the community. A majority (78%) of the persons reported difficulties using ET. The most common difficulties were related to the use of telecommunication and computers. Despite these difficulties, a majority still used most objects and services independently. Twenty-six participants (72%) perceived that their difficulties using ET influenced their everyday activities and their possibility to participate at home and in the community. The results indicate that rehabilitation following an ABI should consider whether clients' use of ET influences their activity and participation and adopt interventions accordingly. The results also indicate that difficulties using ET need to be considered in the design of community services to prevent societal barriers. PMID- 19903094 TI - Optimizing T-cell expansion: have we reached the limit? PMID- 19903096 TI - Phenotypical/functional characterization of in vitro-expanded mesenchymal stromal cells from patients with Crohn's disease. AB - BACKGROUND AIMS: Because of their capacity to modulate the immune response and promote tissue repair, mesenchymal stromal cells (MSC) represent a potential novel treatment for autoimmune/inflammatory diseases, including Crohn's disease (CD). The aim of the study was in vitro characterization of MSC from active CD patients for future clinical application. METHODS: MSC from the bone marrow (BM) of seven CD patients (median age 32 years) were expanded ex vivo in the presence of 5% platelet lysate; cells were investigated for clonogenic efficiency, proliferative capacity, morphology, immunophenotype, differentiation potential, genetic stability and ability to suppress in vitro proliferation of both autologous and allogeneic lymphocytes to polyclonal mitogens. Results were compared with those of BM MSC of four healthy donors (HD). RESULTS: MSC were successfully expanded from all patients. Colony-forming unit-fibroblast (CFU-F) frequency and proliferative capacity were comparable in CD and HD MSC. CD MSC showed typical spindle-shaped morphology and differentiated into osteoblasts, adipocytes and chondrocytes. Surface immunologic markers did not differ between CD and HD MSC, with the only exception of sizeable levels of HLA-DR at early culture passages [12-84% at passage (P)1] in the former. CD MSC ceased their growth at variable passages (from P8 to P25) and entered senescence without any change in morphology/proliferation rate. Array-comparative genomic hybridization demonstrated that CD MSC do not show imbalanced chromosomal rearrangements. Both CD and HD MSC inhibited in vitro proliferation of lymphocytes to mitogens. CONCLUSIONS: CD MSC show biologic characteristics similar to HD MSC and can be considered for anti-inflammatory and reparative cell therapy approaches in patients with refractory disease. PMID- 19903097 TI - Telomerase activity and expression in adult human mesenchymal stem cells derived from amyotrophic lateral sclerosis individuals. AB - BACKGROUND AIMS: Telomerase is a ribonucleoprotein that maintains the length of telomeres, and thus controls the proliferation and lifespan of cells. Recent studies suggest the involvement of telomerase in the protection of cells from apoptosis. Adult human mesenchymal stromal cells (hMSC) possess the capacity to proliferate and differentiate into a variety of cell types. hMSC derived from a healthy donor lack telomerase activity but their expression has not been investigated in hMSC derived from diseased adults. Cell replacement therapy using adult hMSC has been suggested as a promising therapeutic approach for amyotrophic lateral sclerosis (ALS). Therefore, we characterized the telomerase activity and expression in hMSC derived from bone marrow (BM) of ALS patients and compared them with those derived from a healthy donor. METHODS: Telomerase activity was examined with a TRAP assay and real-time polymerase chain reaction telomerase quantification assay. Telomerase protein was detected by Western blot and immunofluorescence analysis, and telomerase RNA transcripts were identified by Northern blot. RESULTS: Telomerase activity, telomerase enzyme protein and telomerase RNA transcripts were demonstrated in hMSC derived from ALS, but were undetectable in hMSC from the healthy donor. Telomerase activity in the hMSC of ALS patients was 106-fold lower compared with tumor cells. CONCLUSIONS: The detection of telomerase expression in hMSC derived from ALS patients and not a healthy donor suggests a possible role for telomerase in the response of hMSC to the disease. The presence of telomerase expression did not impair the ability of the ALS hMSC to differentiate, suggesting the use of these cells for cytotherapy treatments. PMID- 19903095 TI - Important precautions when deriving patient-specific neural elements from pluripotent cells. AB - Multipotent human neural stem cells (hNSC) have traditionally been isolated directly from the central nervous system (CNS). To date, as a therapeutic tool in the treatment of neurologic disorders, the most promising results have been obtained using hNSC isolated directly from the human fetal neuroectoderm. The propagation ability of such tissue-derived hNSC is often limited, however, making it difficult to establish a large-scale culture. Following engraftment, these hNSC often show low efficiency in generating the desired neuronal cells necessary for reconstruction of the damaged host milieu and, as a result, have failed to give satisfactory results in clinical trials so far. Alternatively, human embryonic stem cells (hESC) offer a pluripotent reservoir for in vitro derivation of a rich spectrum of well-characterized neural-lineage committed stem/progenitor/precursor cells that can, theoretically, be picked at precisely their safest and most efficacious state of plasticity to meet a given clinical challenge. However, the need for 'foreign' biologic additives and multilineage differentiation inclination may make direct use of such cell-derived hNSC in patients problematic. The hNSC, when derived from pluripotent cells under protocols presently employed in the field, tend to display not only a low efficiency in neuronal differentiation, but also an inclination for phenotypic heterogeneity and instability and, hence, increased risk of tumorigenesis following engraftment. For hNSC derived in vitro to be used safely in therapeutic paradigms, it requires conversion of human pluripotent cells uniformly to cells that are restricted to the neural lineage in need of repair. Developing strategies for direct induction of human pluripotent cells exclusively into neural-committed progenies at a broad range of developmental stages will allow a large supply of optimal therapeutic hNSC tailor-made for safe and effective treatment of particular neurologic diseases and injuries in patients. PMID- 19903098 TI - Inhibition of myostatin promotes myogenic differentiation of rat bone marrow derived mesenchymal stromal cells. AB - BACKGROUND AIMS: Mesenchymal stromal cells (MSC) have been thought to be attractive candidates for the treatment of Duchenne muscular dystrophy (DMD), but the rate of MSC myogenesis is very low. Thus MSC treatment for DMD is restricted. Myostatin (Mstn), a negative regulator of myogenesis, is known to be responsible for limiting skeletal muscle regeneration. We hypothesized that inhibition of Mstn by using anti-Mstn antibody (Ab) would ameliorate the myogenic differentiation of MSC in vitro and in vivo. METHODS: MSC were isolated from rat bone marrow. Induced rat MSC (rMSC) were treated with various concentrations of anti-Mstn Ab. The expression of myogenic differentiation antigen (MyoD), myogenin and myosin heavy chain-type alpha (MHC-alpha) were estimated by immunofluorescence analysis and reverse transcription-polymerase chain reaction (RT-PCR). Adipogenic differentiation of rMSC inhibited by anti-Mstn Ab was evaluated by Oil Red O staining. The expression of dystrophin was detected 16 weeks after anti-Mstn Ab injection and rMSC transplantation by immunofluorescence staining, RT-PCR and Western blot. Motor function, serum creatine kinase (CK) and histologic changes were also evaluated. RESULTS: Five-azacytidine-mediated myogenic differentiation induced significant endogenous Mstn expression. Anti Mstn Ab improved the expression of MyoD, myogenin and MHC-alpha and inhibited adipocyte formation. Sixteen weeks after transplantation, the inhibition of Mstn had improved motor function and muscle mass. In accordance with the increased motor function and muscle mass, dystrophin expression had increased. Furthermore, serum CK and centrally nucleated fiber (CNF) levels decreased slightly, suggesting specific pathologic features of the dystrophic muscle were partially restored. CONCLUSIONS: Using anti-Mstn Ab, we found that inhibition of Mstn improved myogenic differentiation of rMSC in vitro and in vivo. A combination of Mstn blockade and MSC transplantation may provide a pharmacologic and cell-based strategy for the treatment of DMD. PMID- 19903099 TI - Labeling and tracking of mesenchymal stromal cells with EdU. AB - BACKGROUND AIMS: The thymidine analog bromodeoxyuridine (5-bromo-2-deoxyuridine; BrdU) has been used widely to label cells in culture and in tissue. The labeled cells can also be tracked when transplanted into a suitable host. In the present study we tested a new thymidine analog, 5-ethynyl-2-deoxyuridine (EdU), for labeling and tracking of mesenchymal stromal cells (MSC), specifically adipose tissue-derived stem cells (ADSC). METHODS: Labeling of ADSC was examined for the dosage effect of EdU and stability of label by Alexa-594 staining followed by fluorescence microscopy. Labeling of various organs/tissues was done by intraperitoneal injection of EdU and examined by histology and fluorescence microscopy. Tracking of ADSC was done by intratissue or intravenous transplantation of EdU-labeled ADSC into various tissues and examined by histology and fluorescence microscopy. RESULTS: EdU was incorporated specifically into the nucleus in approximately 50% of ADSC and the percentage of cells that remained fully labeled declined with time. Peritoneal injection of EdU resulted in the appearance of EdU-positive cells in most organs and tissues. In the intestine, EdU-positive cells were found in both the epithelium and connective tissues 7 h after injection. Long-term (2-6 week) follow-ups found EdU-positive cells only in the connective tissue. Tracking of ADSC was successful in tissues 10 weeks after intratissue or intravenous transplantation. CONCLUSIONS: Cell labeling with EdU in culture or living animals can be performed easily. The detection of EdU label requires no harsh treatment or immunologic reaction, as detection of BrdU label does. EdU can be used for long-term tracking of ADSC. PMID- 19903100 TI - Properties and growth of human bone marrow mesenchymal stromal cells cultivated in different media. AB - BACKGROUND AIMS: Human mesenchymal stromal cells (hMSC) are a promising tool for future clinical application, but their use requires rapid cell expansion in media suitable for clinical use. Therefore, we tested the influence of several culture media on colony formation, population doubling (PD) time, cell cycle and surface marker expression. METHODS: hMSC isolated from human bone marrow (BM) obtained from healthy donors were seeded and expanded in different culture media: alpha minimum essential medium (MEM) supplemented with 2.5%, 5%, 10% or 20% fetal bovine serum (FBS), 5% or 10% human cord blood serum (hCBS), 5% or 10% human blood serum from AB adult donors (hABS), or mesenchymal stem cell growth medium (MSCGM). The number, diameter and total area of the colonies formed and PD time were determined, and the cell cycle and 16 surface markers were analyzed. RESULTS: Colony-forming efficiency was best in alpha-MEM/hCBS and alpha-MEM/hABS, good in MSCGM and worst in alpha-MEM/FBS. The shortest PD time was achieved in media enriched with human sera or MSCGM, while the time was increased in alpha MEM/FBS. The largest proliferating fraction was seen in MSCGM followed by media enriched with human sera; the fraction was smallest in alpha-MEM/FBS. Staining for CD34, CD45, CD235a and CD271 was negative, while that for CD29, CD44, CD73, CD90, CD105 and human leukocyte antigen (HLA)-A, -B, -C was positive in all media tested. Media with human serum did not adversely affect the differentiation potential of hMSC, and differentiation into osteoblasts was enhanced. CONCLUSIONS: The choice of serum influences hMSC expansion and cell properties; alpha-MEM supplemented with hABS seems to be a promising candidate for clinical use. PMID- 19903102 TI - Ex vivo-expanded autologous bone marrow-derived mesenchymal stromal cells in human spinal cord injury/paraplegia: a pilot clinical study. AB - BACKGROUND AIMS: Spinal cord injury (SCI) is a medically untreatable condition for which stem cells have created hope in the last few years. Earlier pre clinical reports have shown that transplantation of bone marrow (BM) mesenchymal stromal cells (MSC) in SCI-simulated models can produce encouraging results. In a clinical pilot study, we investigated the growth kinetics of BM MSC from SCI patients, their safety and functional improvement post-transplantation. METHODS: Thirty patients with clinically complete SCI at cervical or thoracic levels were recruited and divided into two groups based on the duration of injury. Patients with <6 months of post-SCI were recruited into group 1 and patients with >6 months of post-SCI were included into group 2. Autologous BM was harvested from the iliac crest of SCI patients under local anesthesia and BM MSC were isolated and expanded ex vivo. BM MSC were tested for quality control, characterized for cell surface markers and transplanted back to the patient via lumbar puncture at a dose of 1 x 10(6) cells/kg body weight. RESULTS: At the time of writing, three patients had completed 3 years of follow-up post-BM MSC administration, 10 patients 2 years follow-up and 10 patients 1 year follow-up. Five patients have been lost to follow-up. None of the patients have reported any adverse events associated with BM MSC transplantation. CONCLUSIONS: The results indicate that our protocol is safe with no serious adverse events following transplantation in SCI patients. The number of patients recruited and the uncontrolled nature of the trial do not permit demonstration of the effectiveness of the treatment involved. However, the results encourage further trials with higher doses and different routes of administration in order to demonstrate the recovery/efficacy if any, in SCI patients. PMID- 19903101 TI - Individual and combined effects of mesenchymal stromal cells and recombinant stimulatory cytokines on the in vitro growth of primitive hematopoietic cells from human umbilical cord blood. AB - BACKGROUND AIMS: We have previously characterized the in vitro growth of two cord blood-derived hematopoietic cell populations in liquid cultures supplemented with recombinant cytokines. In the present study, we assessed the effects of bone marrow-derived mesenchymal stromal cells (MSC) on the growth of such cells. METHODS: CD34(+) CD38(+) Lin(-) and CD34(+) CD38(-) Lin(-) cells were obtained by negative selection, and cultured in the presence of marrow-derived MSC and/or early- and late-acting cytokines. Hematopoietic cell growth was assessed throughout a 30-day culture period. RESULTS: In the presence of MSC alone, both populations showed significant proliferation. Direct contact between MSC and CD34(+) cells was fundamental for optimal growth, especially for CD34(+) CD38(-) Lin(-) cells. In the presence of early-acting cytokines alone, cell growth was significantly higher than in cultures established with MSC but no cytokines. In cultures containing both MSC and early-acting cytokines, a further stimulation was observed only for CD34(+) CD38(-) Lin(-) cells. The cytokine cocktail containing both early- and late-acting cytokines was significantly more potent at inducing hematopoietic cell growth than the early-acting cytokine cocktail. When cultures were supplemented with early- and late-acting cytokines, MSC had no further effect on the growth of hematopoietic cells. CONCLUSIONS: MSC seem to play a key role, particularly on more primitive (CD34(+) CD38(-) Lin(-)) cells, only in the absence of cytokines or the presence of early-acting cytokines. When both early- and late-acting cytokines are present in culture, MSC seem to be unnecessary for optimal development of CFC and CD34(+) cells. PMID- 19903103 TI - Optimization and validation of a robust human T-cell culture method for monitoring phenotypic and polyfunctional antigen-specific CD4 and CD8 T-cell responses. AB - BACKGROUND AIMS: Monitoring cellular immune responses is one prerequisite for the rational development of cancer vaccines. METHODS: We describe an extensive effort to optimize and validate quantitatively an in vitro T-cell culture method by determining the phenotype and function of both CD4(+) and CD8(+) T cells, including measurement of the phenotype markers CCR7, CD45RA, CD28 and CD27 and the functional markers interferon (IFN)-gamma, interleukin (IL)-2, macrophage inflammatory protein (MIP)-1beta, tumor necrosis factor (TNF)-alpha and CD107a. RESULTS: Autologous peripheral blood mononuclear cells (PBMC) were potent stimulators that expanded antigen (Ag)-specific CD8(+) T cells during short-term culture with the addition of IL-2 and IL-15 cytokines. Polyfunctional Ag-specific CD4(+) and CD8(+) T cells were detectable using this method. CONCLUSIONS: Our culture system represents a robust human T-cell culture protocol that permits phenotypic, quantitative and qualitative evaluation of vaccine-induced CD4(+) and CD8(+) T-cell responses. PMID- 19903104 TI - Efficient clinical-scale enrichment of lymphocytes for use in adoptive immunotherapy using a modified counterflow centrifugal elutriation program. AB - BACKGROUND AIMS: Clinical-scale lymphocyte enrichment from a leukapheresis product has been performed most routinely using costly magnetic bead separation systems that deplete monocytes, but this procedure may leave behind residual beads or antibodies in the enriched cell product. Counterflow centrifugal elutriation has been demonstrated previously to enrich monocytes efficiently for generation of dendritic cells. This study describes a modified elutriation procedure for efficient bead-free economical enrichment of lymphocytes from leukapheresis products from healthy donors and study subjects with human immunodeficiency virus (HIV) infection or malignancy. METHODS: Modified program settings and conditions for the CaridianBCT Elutra device were investigated to optimize lymphocyte enrichment and recovery. Lymphocyte enrichment was measured using a novel approach utilizing cell sizing analysis on a Beckman Coulter Multisizer and confirmed by flow cytometry phenotypic analysis. RESULTS: Efficient enrichment and recovery of lymphocytes from leukapheresis cell products was achieved using modified elutriation settings for flow rate and fraction volume. Elutriation allowed for enrichment of larger numbers of lymphocytes compared with depletion of monocytes by bead adherence, with a trend toward increased lymphocyte purity and yield via elutriation, resulting in a substantial reduction in the cost of enrichment per cell. Importantly, significant lymphocyte enrichment could be accomplished using leukapheresis samples from healthy donors (n=12) or from study subjects with HIV infection (n=15) or malignancy (n=12). CONCLUSIONS: Clinical-scale closed-system elutriation can be performed efficiently for the selective enrichment of lymphocytes for immunotherapy protocols. This represents an improvement in cost, yield and purity over current methods that require the addition of monocyte-depleting beads. PMID- 19903105 TI - Impact of culture medium on the expansion of T cells for immunotherapy. AB - BACKGROUND AIMS: Encouraging evidence of clinical benefits from cancer immunotherapy is beginning to accumulate in several clinical trials. Cancer immunotherapy is based on two main methods, active vaccination and cell-transfer therapy. The ex vivo expansion of T cells is required to monitor vaccine-induced antigen-specific T cells or prepare large numbers of reactive lymphocytes for adoptive transfer. METHODS: We examined the influence of culture medium on T-cell growth, cytotoxicity and phenotype after activation using immobilized anti-CD3 monoclonal antibody or Zoledronate stimulation. Peripheral blood mononuclear cells (PBMC) were cultured in RPMI, AIM-V or OpTmizer with or without autologous serum. RESULTS: When supplemented with sufficient serum, RPMI was a good culture medium for T-cell expansion following anti-CD3 stimulation. Addition of autologous serum to AIM-V or OpTmizer increased the numbers of cells obtained to a similar extent, but their phenotype and function were quite different. Activated T cells cultured with OpTmizer mediated greater cytotoxicity than any other culture. Regardless of the media used, the main population expanded after CD3 stimulation was CD3(+) CD8(+). While more CD3(+) CD4(+) T cells were induced in RPMI and AIM-V, more CD3(-) CD56(+) cells and CD3(+) CD56(+) T cells were induced in OpTmizer. When cells were stimulated by Zoledronate for 14 days, approximately 7.2 times and 11.5 times more gammadelta T cells were obtained in OpTmizer than AIM-V or RPMI, respectively. CONCLUSIONS: Successful immunotherapy depends on the selection of appropriate culture media to support efficient expansion of the type of T cell desired. PMID- 19903106 TI - Harvesting human adipose tissue-derived adult stem cells: resection versus liposuction. AB - BACKGROUND: Adipose tissue is an abundant source of mesenchymal stem cells (MSC), which can be used for tissue-engineering purposes. The aim of our study was to determine the more suitable procedure, surgical resection or liposuction, for harvesting human adipose tissue-derived stem cells (hASC) with regard to viability, cell count and differentiation potential. METHODS: After harvesting hASC, trypan blue staining and cell counting were carried out. Subsequently, hASC were cultured, analyzed by fluorescence-activated cell sorting (FACS) and differentiated under adipogenic, osteogenic and chondrogenic conditions. Histologic and functional analyzes were performed at the end of the differentiation period. RESULTS: No significant difference was found with regard to the cell counts of hASC from liposuction and surgically resected material (P=0.086). The percentage of viable cells was significantly higher for liposuction aspirates than for resection material (P=0.002). No significant difference was found in the adipogenic differentiation potential (P=0.179). A significantly lower number of cultures obtained from liposuction material than from resection material could be differentiated into osteocytes (P=0.049) and chondrocytes (P=0.012). DISCUSSION: Even though some lineages from lipoaspirated hASC can not be differentiated as frequently as those from surgically resected material, liposuction may be superior for some tissue-engineering purposes, particularly because of the less invasive harvesting procedure, the higher percentage of viable cells and the fact that there is no significant difference between lipoaspirated and resected hASC with regard to adipogenic differentiation potential. PMID- 19903107 TI - Serum-free, xeno-free culture media maintain the proliferation rate and multipotentiality of adipose stem cells in vitro. AB - BACKGROUND AIMS: Human adipose stem cells (ASC) are an abundant, readily available population of multipotent progenitor cells that reside in adipose tissue. ASC have been shown to have therapeutic applicability in pre-clinical studies, but a standardized expansion method for clinical cell therapy has yet to be established. Isolated ASC are typically expanded in medium containing fetal bovine serum (FBS); however, sera and other culturing reagents of animal origin in clinical therapy pose numerous safety issues, including possible infections and severe immune reactions. METHODS: To identify optimal conditions for ex vivo expansion of ASC, the effects of seven serum-free (SF) and xeno-free (XF) media were investigated with both FBS and allogeneic human serum (alloHS; as a control media). Surface marker expression, proliferation, morphology and differentiation analyzes were utilized for investigating the effects of media on ASC. RESULTS: The proliferation and morphology analysis demonstrated significant differences between ASC cultured in SF/XF culture media compared with serum-containing culture media, with medium prototype StemPro MSC SFM XenoFree providing significantly higher proliferation rates than ASC cultured in media containing serum, while still maintaining the differentiation potential and surface marker expression profile characteristic of ASC. CONCLUSIONS: Looking forward, fully defined XF media formulations will provide the means for the development and approval of safer clinical cell therapy treatments. However, to fully recognize the capacity of these XF culture media, further pre-clinical safety and efficacy studies must be performed. PMID- 19903108 TI - Recurrent incarceration and/or sacculation of the gravid uterus: a review. AB - Retroverted uterine incarceration with sacculation of the anterior wall is reported to occur approximately in 1/3000 pregnancies. A literature search identified only one case report of incarceration of an anteflexed gravid uterus and six reported cases of recurrent incarceration and/or sacculation. We present a case of an incarceration of an anteflexed uterus in the first pregnancy, followed by a retroflexed incarceration in the second pregnancy. From this, a review is presented on recurrent uterine incarceration and/or sacculation. PMID- 19903109 TI - Pregnancy complications in women with heart disease. AB - OBJECTIVE: To identify pregnancy complications of women with heart disease delivering at a university hospital. METHODS: A retrospective study was carried out of 193 pregnant women with heart disease delivered at a university hospital between January 1997 and December 2006. RESULTS: Rheumatic heart disease (RHD), congenital heart disease (CHD), arrhythmia and cardiomyopathy were observed in 133 (68.9%), 26 (13.5%), 32 (16.6%) and 2 (1%) cases, respectively. Obstetric complication was found in 27 (14%) cases that was composed of preterm delivery (11.4%), gestational diabetes (1%), pregnancy induced hypertension (1%) and postpartum hemorrhage (0.5%). Cardiac complication was observed in 24 (12.4%) cases. Congestive heart failure was the most common cardiac complication which observed in 11 (5.7%) cases. There were four (2.1 %) maternal deaths, three cases in CHD group and one case in RHD group. Preterm infant was observed in 22 (11.4%) cases. Thirteen percent had low birth weight and 8.3% were small for gestational age. There were no perinatal deaths or congenital anomalies. CONCLUSION: Form this study, RHD with pregnancy is still predominant. The most common obstetric complication was preterm delivery. The most common cardiac complication was congestive heart failure. PMID- 19903110 TI - Relationship between brainstem auditory function during the neonatal period and depressed Apgar score. AB - OBJECTIVE: To understand whether infants with depressed Apgar scores are at high risk of brainstem auditory impairment, we examined the relationship between brainstem auditory function during the neonatal period and depressed Apgar score. METHODS: Brainstem auditory evoked responses (BAERs) were recorded from day 1 to day 30 in 145 term infants with Apgar scores < or = 6. RESULTS: For 1-min Apgar score none of BAER response wave latencies and interpeak intervals during the first 15 days correlated significantly with the score. On day 30, all wave latencies but no intervals correlated negatively with the score (all p < 0.05). For 5-min Apgar score, only on day 3 wave V latency and I-V and III-V intervals correlated negatively with the score (all p < 0.05). For 10-min Apgar score, only III-V interval correlated negatively with the score (p < 0.05). No correlation was found between BAER variables on any other days and 5 and 10-min Apgar scores. Wave V latency and I-V and III-V intervals on day 3 were all significantly longer in infants with 5-min Apgar scores < or = 6 than in those with scores > 6. CONCLUSIONS: During the neonatal period, only on day 3 after birth a depressed 5 min Apgar score is an indicator associated with central auditory impairment. A depressed 1-min score may be associated with later peripheral auditory impairment. PMID- 19903112 TI - Mass closure of visceral peritoneum at cesarean section. A proposal method. AB - The non-closure of the visceral peritoneum (VP) is the gold standard in caesarean section (CS). For surgeons convinced of the benefits of VP closuring, we propose an alternative method for VP closure during CS, named by us, "Mass Closure Method" (MCM). According to our experience, this method has already been experimented, as described earlier, in 314 CSs in loco-regional anaesthesia. All CSs in our department are performed using the modified Joel Cohen incision in the Stark CS, with the Munro Kerr way of hysterotomy. During haemostasis in 18 cases, author observed a small sub-peritoneal haematoma (5.7%), solved intra-operatively by singular stitches; 10 women received a bladder flap intraoperative detachment, with an accurate haemostasis (3.1%). No case of bladder flap haematoma was recorded, but only hyperechogenic areas in the vesicouterine space (VUS), to be ascribed to the intra-operatives stitches. The direct incision, no bladder flap formation and not suturing a part of the VP, substantially reduce the VUS surface, consequently decrease bleeding, and prevent a pouch formation. On the basis of the author's experiences, the MCM could represent for obstetrics a safe alternative to the surgical approach during CS, as it cut down operative time, surgical bleeding, BHF formation for the non-bladder flap formation, thus reducing the VUS surface. PMID- 19903111 TI - Consequences of methemoglobinemia in pregnancy in newborns, children, and adults: issues raised by new findings on methemoglobin catabolism. AB - OBJECTIVE: The aim of this review is to warn about the effects of methemoglobin and its catabolic products and the toxic effects caused by environmental oxidants that cause high-risk pregnancy and may later impair the health of newborns, children and adolescents. METHODS: In our study of pregnant women (n = 36) whose methemoglobin level was >1.5 g/l, we took blood samples from their newborns to determine the frequency of sister chromatid exchange (SCE) by cultivating lymphocytes. The research took place at the Department of Biology and Medical Genetics of the School of Medicine in Rijeka (Croatia). RESULTS: The results have shown that no deviation in the SCE frequency was found in either case (1990). We examined data on the health of newborns collected at perinatal hospital departments in Rijeka (Croatia), the preschool office and school service at the Labin Health Center and continued until they were 18 years of age (2008). The statistics obtained by applying the chi-square test show that the incidences of neonatal jaundice (p = 0.034), heart murmur at a later age (p = 0.011) and dyslalia and learning/memory impairments (p = 0.002) were significantly higher than in children of control mothers (n = 19). CONCLUSION: Depending on the mother's exposure to environmental oxidants, during its development the fetus is more likely to be affected by methemoglobin and hemolysis. Oxidants affect the vascular endothelium of kidneys, brain and other vital organs, because they have the capacity to cross the damaged fetomaternal placental barrier. 'Fetal preeclampsia' is an expected manifestation of the condition. Our research proves our thesis on the pathophysiological relationship between methemoglobinemia and unexplained jaundice and hyperbilirubinemia, heart murmur at a later age, dyslalia and learning and memory impairments that have not exactly been demonstrated yet. PMID- 19903113 TI - Sexually transmitted infection is associated with increased risk of preterm birth in South Carolina women insured by Medicaid. AB - OBJECTIVE: We investigated the association between diagnosed maternal sexually transmitted infections (STIs) and very preterm or late preterm birth in 108,346 South Carolina women receiving medical care purchased by Medicaid from 1996 through 2002, and delivering singleton infants. METHODS: ICD9 codes in South Carolina Medicaid data were used to identify maternal infections. Gestational age was obtained from birth certificates. RESULTS: Trichomoniasis, gonorrhea, and/or Chlamydia/non-gonococcal urethritis (NGU) were diagnosed in 4,208 women. Preterm birth occurred in just over 9% of pregnancies. Women diagnosed with one of the infections in the first 7 months of pregnancy were significantly more likely to deliver very preterm infants (adjusted HR = 1.73, p < 0.0001). Women with infection in the first 8 months were significantly more likely to deliver between 33 and 36 weeks (adjusted HR = 1.39, p < 0.0001). Of the three infections, Chlamydia/NGU was most strongly associated with preterm birth and infections occurring later in pregnancy (months 6, 7, and 8). CONCLUSION: STIs appear to be preventable risk factors for very and late preterm birth. PMID- 19903114 TI - Vaginal fetal fibronectin evaluation before and immediately after ultrasonographic vaginal cervical length measurements in symptomatic women at risk of preterm birth: a pilot study. AB - OBJECTIVE: To determine if vaginal ultrasound for cervical length measurement induces the release of vaginal fetal fibronectin (fFN), leading to a false positive fFN test. METHODS: Participants included women with singleton pregnancies at 24-34 weeks' gestation who presented with uterine contractions without bleeding or membrane rupture. Women who had had intercourse or underwent pelvic examination less than 24 h previously were excluded. The first fFN test was followed immediately by vaginal ultrasonography with a transvaginal probe (three images per patient) and, thereafter, a second fFN test. RESULTS: The first fFN test was positive in three patients, and in all, the second, post-ultrasound, fFN test was also positive. In all women with a negative baseline fFN test (n = 25), the second, post-ultrasound, fFN test was also negative. CONCLUSION: Vaginal ultrasound examination does not artificially change the fFN status. This is in contrast to common understanding and may permit the performance of ultrasound examination before fFN, which can be restricted to cases of short cervix. PMID- 19903115 TI - Maternal obesity, gestational hypertension, and preterm delivery. AB - OBJECTIVE: To study maternal obesity as a risk factor for preterm delivery. METHODS: Maine State Birth Records Database from 1996 through 2006 was evaluated to investigate obese pregnant women compared with normal weight women regarding risk for preterm delivery. Multiple risk factors and outcomes were studied in univariable and multivariable models. RESULTS: Among 58,112 pregnant women, 8% (n = 4653) gave birth to preterm infants. Univariable analyses revealed a relationship between obesity and increased risk of prematurity. In multivariable regressions, the most important intermediate variable appears to be gestational hypertension/preeclampsia. CONCLUSIONS: As maternal body mass index increases in pregnancy, the risk of preterm delivery and other maternal complications increases. The obesity-prematurity relationship is complex, with hypertensive disorders of pregnancy playing a crucial role. More detailed analyses of causal pathways are warranted. PMID- 19903116 TI - Living with a prostate cancer diagnosis: a qualitative 2-year follow-up. AB - AIM: Previous research has identified how newly diagnosed prostate cancer affects men's daily lives, including daily activities and existential issues. The aim of this qualitative study was to provide information if and how prostate cancer affects men's daily lives 2 years after the diagnosis. METHODS: A second follow up interview with men who were diagnosed with localized or advanced prostate cancer approximately 18-24 months earlier. Twenty-two men aged 50-85 years participated, data were analyzed by hermeneutical interpretation with Gadamer's approach. RESULTS: The men feel healthy, but prostate cancer affects their daily lives. They experience every day fatigue associated with several changes in life due to age. Three equivalent fusions which influenced the men are: 'Age is claiming its due', 'Living with uncertainty', and 'Strengthen self-esteem'. The unifying fusion is identified as 'Balancing a changed life situation.' CONCLUSIONS: There is need for knowledge and guidance for men with prostate cancer on how to adapt to new life situation decreasing uncertainty and increasing welfare. Men found a sense of pride, despite physical changes, in appreciating independence and close relationships. Value of life was readjusted. The findings indicate harmony in living, enjoying being frank as well as rating the 'little' things. PMID- 19903117 TI - Economics of assisted reproduction: access to fertility treatments and valuing live births in economic terms. AB - The intricate relationship between economic conditions and natural fertility is known to influence both the timing and number of children conceived. For infertile couples, the relationship between economics and fertility is more explicit because of the necessity for many couples to pay for treatment to achieve childbirth. Consequently, affordability often dictates whether or not someone is able to undergo treatment, as well as the types of treatments available. Economics can also be used to describe treatment outcomes achieved through the use of fertility treatments. While gynaecologists and couples speak of outcomes in terms of live births, economists are often inclined to view live births and their influence on society in economic terms. In this review we consider two distinct elements of economics and assisted reproduction. Firstly, how economics (i.e. affordability) can influence demand for, and access to, fertility treatments, and secondly, how methods for valuing live births achieved using assisted reproductive technologies in economic terms can highlight the importance of these children in the context of ageing populations. This review will attempt to illustrate that the economic benefits attributed to children conceived through fertility treatments are much greater than health costs required for conception and should be considered in future reimbursement decisions in this therapy area. PMID- 19903118 TI - Insulin resistance and overweight-obese women with polycystic ovary syndrome. AB - AIM: To estimate (1) the prevalence of insulin resistance (IR) by fasting glucose: insulin ratio (G:I) (G:I or= 23 kg/m(2)) were studied. PCOS was diagnosed as per the Rotterdam 2003 criteria. BMI, abdominal circumference (AC), hirsutism (Ferriman Gallway score >or=6), presence of acne and acanthosis nigricans (AN) were noted in each case. Serum testosterone, sex hormone binding globulin (SHBG), fasting plasma glucose and insulin levels were measured. RESULTS: 23.5% women were found to have IR. There were no differences in age, frequency of hirsutism, acne, serum testosterone and fasting glucose levels between insulin-resistant and insulin-sensitive women. However, there were significant differences in BMI, AC, frequency of AN, SHBG levels, fasting insulin levels and FAI between the two groups. CONCLUSION: PCOS women with IR are more obese; they have more upper body adiposity and AN. They are more hyperandrogenic. Simple clinical parameters will help to suspect IR in PCOS women. PMID- 19903119 TI - Endometriotic epithelial cells induce MMPs expression in endometrial stromal cells via an NFkappaB-dependent pathway. AB - OBJECTIVE: To explore the stroma-epithelium interactions in endometriosis and to identify the possible signalling pathways involved in this cross-talk. DESIGN: Laboratory study via primary cultured endometrial stromal and epithelial cells. SETTING: University Hospital. PATIENTS: Fifteen patients with endometriosis confirmed by histopathology were recruited in the study, and 12 women free of endometriosis were used as control group. INTERVENTION(S): Specific NFkappaB inhibitor 1-Pyrrolidinecarbodithioic acid ammonium salt (PDTC) was used in cell cultures. MAIN OUTCOME MEASURE(S): The expression and secretion of MMP-2, MMP-9, TIMP-1, TIMP-2 and the DNA-binding activity of NFkappaB in normal endometrial stromal cells or in co-cultures with normal or endometriotic epithelial cells from patients with endometriosis. RESULT(S): Endometrial epithelial cells induced MMP-9 and MMP-2 expression in normal stromal cells in vitro. In co-cultures with endometriotic epithelial cells, normal endometrial stromal cells expressed and secreted higher MMP-2 (p < 0.05) and MMP-9 (p < 0.05). Specific inhibition of NFkappaB pathway in stromal cells abolished this induction effect by epithelial cells. CONCLUSION(S): Endometriotic epithelial cells induce MMPs expression and secretion in normal endometrial stromal cells via an NFkappaB-dependent pathway in vitro. This cross-talk between epithelial cells and stromal cells may facilitate the implantation and extension of the ectopic foci and favour the development of the disease. PMID- 19903121 TI - Interaction between persons with profound intellectual and multiple disabilities and their partners: a literature review. AB - BACKGROUND: High quality interactions are of crucial importance for quality of life of persons with profound intellectual and multiple disabilities (PIMD). This literature review describes and synthesises studies addressing the interaction between persons with PIMD and their partners. METHOD: A computerised literature search using defined inclusion criteria yielded 15 articles. RESULTS: The literature analysis revealed four components important in interactions: sensitive responsiveness, joint attention, co-regulation, and an emotional component. The abilities and disabilities, interactive behaviours, and personality of persons with PIMD influence these interactions. Additional influences are the partners' interactive strategies, knowledge, and perceptions and the context of the interaction. CONCLUSIONS: An overview model integrates the results and forms a vehicle to facilitate our understanding of interactions with persons with high support needs. Methodological analyses of the studies show lacunae in current research. This review offers a starting point to guide future research and intervention. PMID- 19903124 TI - Systematic review of the effectiveness of mirror therapy in upper extremity function. AB - PURPOSE: This review gives an overview of the current state of research regarding the effectiveness of mirror therapy in upper extremity function. METHOD: A systematic literature search was performed to identify studies concerning mirror therapy in upper extremity. The included journal articles were reviewed according to a structured diagram and the methodological quality was assessed. RESULTS: Fifteen studies were identified and reviewed. Five different patient categories were studied: two studies focussed on mirror therapy after an amputation of the upper limb, five studies focussed on mirror therapy after stroke, five studies focussed on mirror therapy with complex regional pain syndrome type 1 (CRPS1) patients, one study on mirror therapy with complex regional pain syndrome type 2 (CRPS2) and two studies focussed on mirror therapy after hand surgery other than amputation. CONCLUSIONS: Most of the evidence for mirror therapy is from studies with weak methodological quality. The present review showed a trend that mirror therapy is effective in upper limb treatment of stroke patients and patients with CRPS, whereas the effectiveness in other patient groups has yet to be determined. PMID- 19903125 TI - Classification of employment factors according to the International Classification of Functioning, Disability and Health in patients with neuromuscular diseases: a systematic review. AB - PURPOSE: A systematic evaluation of the literature to identify health and contextual factors associated with employment in patients with neuromuscular diseases (NMD) and to perform a best evidence synthesis, taking into account the design of studies, methodological quality and the statistical significance of findings. METHOD: Publications were retrieved by a computerised search in medical and psychological databases. Two reviewers assessed titles and abstracts first and assessed the quality of the remaining full text publications independently as well. Of the residual publications, health and contextual factors associated with employment in patients with NMD were extracted. The factors found were included in a recently developed expanded International Classification of Functioning, Disability and Health scheme. RESULTS: Six hundred and sixty-two titles and abstracts were screened. The main reason to exclude a title and/or abstract was the absence of the study population selected: Facioscapulohumeral Muscular Dystrophy (FSHD), Hereditary Motor and Sensory Neuropathy (HMSN) & Myotonic Dystrophy (MD). Of the remaining 20 full-text publications, eight publications fulfilled the inclusion criteria: two repeated survey designs and six cross sectional studies. Factor extraction resulted in 94 factors related to employment. Ten factors in five publications were indicative for an association with employment status: Disease related factors HMSN, MD & NMD in general), factors related to functions (physical functions, muscle power functions), general personal factors (age, gender and education), work related personal factors (type of occupation, and expressed interest in employment by patients with NMD). CONCLUSION: In the best evidence synthesis ten factors were indicative for an association with employment status in patients with NMD in five publications with good to excellent methodological quality. PMID- 19903126 TI - The move towards community-based rehabilitation in industrialised countries: are we equipped for the challenge? AB - BACKGROUND: Recent challenges to health systems in industrialised countries (e.g., health trends, workforce shortages, geographical dispersion, changing demographics and the growing demand for hospital beds) have prompted a rise in popularity of services loosely labelled community-based rehabilitation (CBR). The rise of CBR is based on the assumption that these models of service delivery have the potential to address some of these challenges by promoting efficient use of community resources. However, due to the way in which CBR has evolved in industrial countries, there is considerable ambiguity surrounding the concept, and even more uncertainty about the methods by which its implementation can be fostered. PURPOSE: To explore the CBR in an industrialised country and the implications of its implementation for the health workforce, health systems and service delivery. METHOD: This article reviews existing literature to explore the concept of CBR as it is applied in industrialised countries. It examines the possible implications of adopting CBR into health systems, including the need for conceptual clarity, a competency frameworks and ongoing professional development. CONCLUSION: This article has shown that for CBR is to become a viable model for the delivery of health services in industrialised countries, a competency framework is needed, together with strong leadership to facilitate the translation of theory into practice. Further, collaboration is required among practitioners, policy makers, unions, consumers, educators and professional associations to support this transformation. PMID- 19903127 TI - Employment of persons with spinal cord lesions injured more than 20 years ago. AB - PURPOSE: The primary objective was to study factors influencing post-injury employment and withdrawal from work in persons who sustained traumatic spinal cord injury (SCI) more than 20 years ago. A secondary objective was to study life satisfaction in the same patients. METHOD: A cross-sectional study with retrospective data of 165 SCI-patients admitted to Sunnaas Rehabilitation Hospital 1961-1982. Multiple logistic regression was used to identify predictors for obtaining work post-injury. A Cox proportional hazards regression model was used to study factors influencing early withdrawal from work, i.e. time from injury until discontinuing employment. RESULTS: Sixty-five percent of the participants were employed at some point after the injury. Thirty-five percent still had work at the time of the survey. The odds of obtaining work after injury were higher in persons of younger age at injury, higher in males versus females, higher for persons with paraplegia versus tetraplegia, and for persons classified as Frankel D-E compared to a more severe SCI. Factors associated with shorter time from injury until discontinuing employment were higher age at injury, incidence of injury after 1975 versus before, and a history of pre-injury medical condition(s). Life satisfaction was better for currently employed participants. CONCLUSION: The study indicates a low employment-rate in persons with SCI, even several years after injury. From the results, we suggest more support, especially to persons of older age at injury and/or with a history of pre-injury medical condition(s), to help them to obtain work and sustain employed for more years after injury. PMID- 19903128 TI - Needs assessment of individuals with stroke after discharge from hospital stratified by acute Functional Independence Measure score. AB - PURPOSE: To determine the needs, barriers and facilitators of function in individuals with stroke after discharge from hospital. To examine the results stratified by the patient's acute score (<41, 41-80, >80) on the functional independence measure (FIM). METHOD: This was a cohort study of 209 patients who had been admitted to hospital because of stroke. Patients were interviewed following hospital discharge using a semi-structured interview and asked to complete and return a quantitative closed-ended survey. RESULTS: For most domains, frequencies of needs varied across the FIM groups. Combining all FIM groups, the interview showed needs related to: physical impairments (35%), time for recovery (33%), education (28%), medical advice (25%), therapies and services (21%), social needs (19%) and emotional needs (18%). From the interview, the most frequent barriers were physical impairments (55%) and emotional concerns (40%). Common facilitators were family support (54%), therapies and medical care (40%) and personal attitudes (22%). Additional needs from the survey concerned: IADL, mobility, ADL, recreation, finances, communication and employment. Additional barriers from the survey were: attitudes, social participation, environments and limited services. CONCLUSIONS: There is a large and varied number of needs and barriers following discharge from hospital that have planning and advocacy implications for rehabilitation teams. PMID- 19903129 TI - Prognostic indicators of functional outcomes in first time documented acute stroke patients following standard dysphagia treatment. AB - PURPOSE: To explore prognostic factors specific to the speech language pathology scope of practice that may be related to the functional outcome of stroke patients with dysphagia. METHOD: The prognostic factors analysed for 100 stroke patients with dysphagia included the swallowing portion of the Functional Assessment Measure (SFAM) at discharge as the dependent variable, and age, severity of stroke, cognitive status, length of stay, amount of individual treatment, and the SFAM, Food Texture, and Liquid Consistencies at admission as the independent variables. Correlation analysis, linear regression and descriptive statistics were used to analyse these variables. RESULTS: Variables that had significant influences on the SFAM discharge levels included the admission ratings of the SFAM, Food Texture, Liquid Consistency, Cognitive FIM levels and length of stay. Amount of individual treatment received was also noted to be significant. Subject age, lesion site and lesion type did not reach significance in correlation or regression analysis. CONCLUSION: Prognostic factors may play an important role in predicting outcome. This data provides speech-language pathologists with knowledge to more effectively communicate the potential outcome of treatment and recovery to patients and families. Additionally, this study stresses the importance of the initial assessment ratings and suggests the need for inter-rater reliability of assessment measures within rehabilitation facilities. PMID- 19903130 TI - Characteristics influencing participation of Australian children with cerebral palsy. AB - PURPOSE: To investigate the extent to which selected individual, family and environmental variables were associated with participation of children who have cerebral palsy in activities outside school. METHODS: Data were gathered through a population-based survey of 114 children born in 1994 or 1995 in Victoria, Australia. Participation was measured using the Children's Assessment of Participation and Enjoyment. Selected independent variables were classified as related to the child, family or environment. Linear regression analysis was used to identify variables associated with participation in informal (activities that require little planning) and formal (those with structure and leaders) activities. RESULTS: Participation in informal activities tended to be greater in children who preferred informal activities and who had higher manual ability (adjusted R(2) = 36.3%). Girls and those with better gross motor function also tended to participate in more activities. The explanatory power of the regression model for participation in formal activities was limited (adjusted R(2) = 4.2%). CONCLUSION: Knowing a child's activity preferences is critical to intervention planning. Being exposed to a range of activities within supportive environments may provide the opportunity to develop preferences, especially in activities where children with cerebral palsy have reduced participation, such as in physical activities. PMID- 19903131 TI - The perceived benefits and barriers to exercise participation in persons with multiple sclerosis. AB - PURPOSE: The purpose of this study was to examine the perceived benefits and barriers to exercise participation in persons with multiple sclerosis (MS). METHOD: A cross-sectional postal survey comprised of 93 adults with MS was conducted. Participants completed the Exercise Benefits and Barriers Scale (EBBS), Spinal Cord Injury Exercise Self-Efficacy Scale (EXSE), Multiple Sclerosis Impact Scale, Disease Steps Scale and International Physical Activity Questionnaire. RESULTS: Forty-three percent of the participants were classified as exercising individuals (EX group) as compared with non-exercising individuals (non-EX group). Participants in the EX group reported significantly higher scores on the EBBS and EXSE. Items related to physical performance and personal accomplishment were cited as the greatest perceived benefits to exercise participation and those items related to physical exertion as the greatest perceived barriers to both the EX and non-EX groups. CONCLUSION: When compared with previous studies conducted in the general population, the participants in the present study reported different perceived barriers to exercise participation. Furthermore, awareness of the benefits of physical activity is not sufficient to promote exercise participation in persons with MS. Perceived exercise self-efficacy is shown to play an important role in promoting exercise participation in persons with MS. PMID- 19903133 TI - Navigating evidence-based information sources in augmentative and alternative communication. AB - An important part of evidence-based practice is to identify the best and most current research evidence to guide clinical practice. The purpose of this paper is to propose the use of the 5-S model by Haynes (2006) as a principled approach for navigating evidence-based information sources related to interventions in augmentative and alternative communication (AAC). The 5-S model is a pyramid that is used from the top down, beginning with systems followed by summaries, synopses, syntheses, and studies. The 5-S model is described along with relevant evidence-based information sources in AAC, and subsequently illustrated with a case scenario. Following the 5-S model may enable practitioners to identify the best and most current research evidence. PMID- 19903134 TI - Translucency and learnability of Blissymbols in Setswana-speaking children: an exploration. AB - Although the importance of iconicity in the learning of symbols has been widely acknowledged, there have been few systematic investigations into the influence of culture on the ratings of symbol iconicity. The purposes of this study were two fold: to determine (a) the translucency ratings of specific Blissymbols as rated by 6- to 7-year-old Setswana-speaking children (one of South Africa's 11 official languages); and (b) whether the ratings changed after second and third exposures in order to determine the learnability of these symbols. This study is partially based on the study by Quist et al. (1998), which utilized Dutch and American participants. Thirty-four Setswana children were exposed to 93 selected Blissymbols. A 3-point semantic differential scale consisting of three faces accompanied each Blissymbol, without the written gloss. This procedure was repeated over a period of 3 days. The results indicated that the majority of Blissymbols were rated as having high translucency ratings. The research further demonstrated significant differences in translucency between first and second exposures, suggesting that learning of the symbols had occurred. The comparison between the results of the current study and the results reported in the Quist et al. study reveal that the translucency ratings of the majority of the selected Blissymbols ranged from moderate to high for all three studies, but that the distribution of symbols across the ratings appears to be different. PMID- 19903135 TI - Spectroscopic and theoretical approaches for the determination of heparin saccharide structure and the study of protein-glycosaminoglycan complexes in solution. AB - Glycosaminoglycans (GAGs), such as heparin and heparan sulphate, are a class of linear, anionic polysaccharides that constitute the carbohydrate component of proteoglycans. The structure of GAG complexes with proteins can reveal details of their mechanisms of action in living systems and help to design new pharmaceuticals. Molecular modelling together with nuclear magnetic resonance (NMR) and other spectroscopic techniques such as circular dichroism (CD) provide indispensable information on structure and dynamics of GAGs and their complexes. The present review focuses on applications of high-resolution NMR, CD and molecular modelling to the analysis of GAGs. The most advanced theoretical methods used at present in GAG research, density functional theory methods (DFT), are also discussed. PMID- 19903136 TI - Gammadelta T lymphocytes producing IFNgamma and IL-17 in response to Candida albicans or mycobacterial antigens: possible implications for acute and chronic inflammation. AB - T lymphocytes bearing the gammadelta T cell receptor are known to play an important role in the first-line defense against viral, bacterial and fungal pathogens. Two main subsets of gammadelta T cells are known, showing distinct functional behaviour: Vdelta2 T lymphocytes, circulating in the peripheral blood, are involved in the response to mycobacterial infections and certain viruses, including coxsakie virus B3 and herpes simplex virus type 2. Vdelta1 T cells are resident in the mucosal-associated lymphoid tissue and are reported to participate in the immunity against Listeria monocytogenes and cytomegalovirus. Vdelta2 T lymphocytes recognize non-peptidic phosphorylated metabolites of isoprenoid biosynthesis, expressed by mycobacteria, while Vdelta1 T cells mainly interact with MHC-related antigens (MIC-A and MIC-B) and with receptors, called UL-16 binding proteins, for the UL-16 protein produced by cytomegalovirus infected cells. Both Vdelta1 and Vdelta2 T cells can produce interferon-gamma in response to MIC-A(+) cells or non-peptide antigens, respectively. Moreover, production of TNF-alpha by human Vgamma9/Vdelta2 T cells has been demonstrated in response to bacterial products and non-peptidic molecules. Recently, it has been reported that gammadelta T lymphocytes can produce IL-17 during Escherichia coli or Mycobacterium tuberculosis infections in mice. This is of interest as IL-17 is emerging as a cytokine crucial in the control of intracellular pathogens and fungi. In this review, we will discuss the possible role of IL-17 producing gammadelta T cells in the regulation of acute and chronic inflammation, focusing on the different response of the two subsets to mycobacterial, viral or fungal antigens. PMID- 19903137 TI - Multifunctional lipoic acid conjugates. AB - Several hundreds of studies published the last decade have reported that alpha lipoic acid (LA) possess the potential to intervene in various therapeutically interesting pathways. However, it should be noted that LA reportedly exerts most of its effects at high micromolar concentrations; that amides of LA exhibit higher biological activity than the parent compound; and that molecular combinations (hybrids) obtained by coupling LA with an amino-substituted bioactive moiety, possess multifunctional activity. The design and synthesis of hybrid molecules encompassing two pharmacophores in one molecular scaffold is a well established approach to the synthesis of more potent drugs with dual activity. Using this approach, various research groups have recently designed and synthesized hybrid compounds with antioxidant activity hyphenated with a wide variety of other activities such as neuroprotective, cardioprotective, anti inflammatory, antidiabetic and anticancer activity as well as enzyme inhibition. Moreover, LA represents an ideal chemical entity for the development of biologically interesting functionalized nanoparticles. Many recent publications describe the use of LA: i) as component of nanospheres and nanoprodrugs, ii) as a linker for the attachment of lipids, carbohydrates, proteins and oligonucleotides on gold nanoparticles to form Self Assembled Monolayers (SAMs) and iii) as surface ligand for cap exchange reactions to prepare water-soluble semiconducting nanocrystal Quantum Dots (QDs). This review is focused on the growing field of the design and synthesis of LA conjugates, for the development of novel molecules with a dual mode of action and the construction of nanosized antioxidants, Self Assembled Monolayers and Quantum Dots. PMID- 19903138 TI - Iron oxide based MR contrast agents: from chemistry to cell labeling. AB - Superparamagnetic iron oxide nanoparticles can be used for numerous applications such as MRI contrast enhancement, hyperthermia, detoxification of biological fluids, drug delivery, or cell separation. In this work, we will summarize the chemical routes for synthesis of iron oxide nanoparticles, the fluid stabilization, and the surface modification of superparamagnetic iron oxide nanoparticles. Some examples of the numerous applications of these particles in the biomedical field mainly as MRI negative contrast agents for tissue-specific imaging, cellular labeling, and molecular imaging will be given. Larger particles or particles displaying a non-neutral surface (thanks to their coating or to a cell transfection agent with which they are mixed) are very useful tools, although the cells to be labeled have no professional phagocytic function. Labeled cells can then be transplanted and monitored by MRI in a broad spectrum of applications. Direct in vivo magnetic labeling of cells is mainly performed by intravenous injection of long-circulating iron oxide-based MRI contrast agents, which can extravasate and/or undergo a cellular uptake in an amount sufficient to allow an MRI visualization of areas of interest such as inflamed regions or tumors. Particles with long circulation times, or able to induce a strong negative effect individually have been also modified by conjugation to a ligand, so that their cellular uptake, or at least their binding to the cell surface, could occur through a specific ligand-receptor interaction, in vivo as well as in vitro. Thus, experimentally as well as in a few trials on humans, iron oxide particles currently find promising applications. PMID- 19903139 TI - Synthesis of thieno[2,3-d]oxazines and thieno[2,3-d]thiazines as subtype specific kainate receptor antagonists. AB - For the development of new antiepileptics the kainate receptors GluR6 and GluR5 are important targets. Based on the anticonvulsant effects of chinazolines and thieno[2,3-d]pyrimidines that are known from the literature, thieno[2,3 d][1.3]oxazines were synthesized and studied for their inhibitory properties at GluR6 and GluR5 receptors. The strongest inhibitor activity was observed with 5 methyl-6-phenyl-thieno[2,3-d][1.3]oxazines with C1 or C3-substituents in position 2 (3b-f). The 2-trihalide-methyl-substituted compounds 3c and 3d were the most active inhibitors at the GluR5-receptor (IC50=23.4 micromol, 16 microl). The 2 isopropyl-substituted compound 3f displayed the strongest activity at the GluR6 receptor (IC(50)=8.7 micromol). A number of thieno[2,3-d][1.3]thiazines and thieno[2,3-d] pyrimidines that were synthesized from the thieno[2,3][1.3]oxazines did not show any activity. PMID- 19903140 TI - Endothelial calcium machinery and angiogenesis: understanding physiology to interfere with pathology. AB - Endothelial cells (ECs) play a pivotal role in physiological and altered tissue neovascularization. They face multiple morphological, biochemical and functional changes during the different phases of angiogenesis, under the regulation of a great number of proangiogenic and antiangiogenic signals, including soluble and insoluble factors, cell-cell and cell-matrix interactions. ECs mutual contacts (and also interactions with other cell types, such as pericytes and smooth vascular muscle cells), motility, proliferation, apoptosis and differentiation are all calcium-dependent events finely tuned in space and time. Most of the angiogenic-related peptidic factors (VEGF, bFGF and others) promote an increase of cytosolic free calcium concentration in ECs, giving rise to calcium-activated intracellular cascades engaged in the different steps of the angiogenic process. A better knowledge of such signals could allow to set new diagnostic and therapeutical approaches aimed to interfere with altered neovascularization, particularly during cancer progression. This review reports the state of the art about endothelial angiogenic-related calcium signaling and discusses the most attractive perspectives for the future. PMID- 19903141 TI - Second messenger modulation: a novel target of future antidepressants? AB - During the transduction of extracellular signals within the cell, the stimulation of specific G protein-coupled receptors (GPCRs) can modulate adenylyl or guanylyl cyclase, phospholipase C activity and ion channels, which regulate second messengers. These, in turn, trigger several biochemical cascades, including Ca(2+) release, activation of protein kinases and gene expression. Significant changes of monoamine GPCR activity may occur in patients suffering from mood disorders and the majority of antidepressants exert part of their effects through GPCR-mediated systems. The main signal transduction pathways activated by metabotropic receptors in the brain and their possible involvement in the pathophysiology of mood disorders will be reviewed herein with a special focus on the horizons opened by this approach in terms of innovative therapeutic strategies. PMID- 19903143 TI - Oxidative stress and mitochondrial dysfunction in atherosclerosis: mitochondria targeted antioxidants as potential therapy. AB - Chronic and acute overproduction of reactive oxygen species (ROS) under pathophysiologic conditions forms an integral part of the development of cardiovascular diseases (CVD), and in particular atherosclerosis. These ROS are released from different sources, such as xanthine oxidase, lipoxygenase, nicotinamide adenine dinucleotide phosphate oxidase, the uncoupling of nitric oxide synthase and, in particular, mitochondria. Endothelial dysfunction, characterized by a loss of nitric oxide (NO) bioactivity, occurs early on in the development of atherosclerosis, and determines future vascular complications. Although the molecular mechanisms responsible for mitochondria-mediated disease processes are not clear, oxidative stress seems to play an important role. In general, ROS are essential to cell function, but adequate levels of antioxidant defenses are required in order to avoid the harmful effects of excessive ROS production. Mitochondrial oxidative stress damage and dysfunction contribute to a number of cell pathologies that manifest themselves through a range of conditions. This review considers the process of atherosclerosis from a mitochondrial perspective, and assesses strategies for the targeted delivery of antioxidants to mitochondria that are currently under development. We will provide a summary of the following areas: the cellular metabolism of reactive oxygen species (ROS) and its role in pathophysiological processes such as atherosclerosis; currently available antioxidants and possible reasons for their efficacy and inefficacy in ameliorating oxidative stress-mediated diseases; and recent developments in mitochondrially-targeted antioxidants that concentrate on the matrix-facing surface of the inner mitochondrial membrane in order to protect against mitochondrial oxidative damage, and their therapeutic potential as a treatment for atherosclerosis. PMID- 19903142 TI - Pharmacomimetics of exercise: novel approaches for hippocampally-targeted neuroprotective agents. AB - Coordinated and constructive physical activity is correlated with the maintenance of cognitive function in humans. Voluntary running also enhances neuroplasticity in adult and aging rodents, but the molecular pathways underlying these effects are still being elucidated. Considering the multifactorial nature of the biochemical links between physical activity and neurophysiology it is likely that there are many pharmacological mechanisms by which the beneficial actions of exercise can be effectively reproduced using chemical agents. Most studies to date have focused on brain-derived neurotrophic factor (BDNF) as a signaling target for the enhancement of neuronal function by exercise. The goal of the current review is to move beyond BDNF by exploring the diversity of molecular pathways regulated by physical activity in a variety of situations. We will discuss the availability and mechanism of action for several diverse physical activity pharmacomimetics. As physical activity enhances both neuroplasticity and cognition, understanding the molecular targets for these effects may lead to the development of protent new therapeutic interventions for age-related neurodegenerative conditions such as Alzheimer's disease. PMID- 19903144 TI - The role of neopterin in atherogenesis and cardiovascular risk assessment. AB - Neopterin is produced by human and primate monocyte/macrophages upon activation by pro-inflammatory stimuli like Th1-type cytokine interferon-gamma. Neopterin has pro-oxidative properties, which have been demonstrated in vitro in physicochemical and cell culture studies and also in in vivo experiments, e.g. the Langendorff perfusion model of rat hearts. In the past several years, the measurement of neopterin concentrations in body fluids including serum, urine and cerebrospinal fluid has revealed a potential role of this molecule in the prediction of long-term prognosis in both patients with cancer and those with systemic infections such as HIV-1 infection. Moreover, elevated neopterin concentrations have been reported in patients with coronary disease compared to controls and in recent years it has become apparent that increased neopterin concentrations are an independent marker for cardiovascular disease and a predictor of future cardiovascular events in patients with coronary artery disease. Current data suggest that the diagnostic performance of neopterin testing is comparable to that of well established biomarkers such as C-reactive protein and cholesterol plasma levels. The present article reviews the role of neopterin in the pathogenesis of cardiovascular disease and as a marker of coronary artery disease progression. PMID- 19903145 TI - Innovations and opportunities to improve conventional (deoxy)nucleoside and fluoropyrimidine analogs in cancer. AB - Many drugs that are currently used for the treatment of cancer have limitations, such as induction of resistance and/or poor biological half-life, which reduce their clinical efficacy. To overcome these limitations several strategies have been explored. Chemical modification by the attachment of lipophilic moieties to (deoxy)nucleoside analogs should enhance the plasma half live, change the biodistribution and improve cellular uptake of the drug. Attachment of a lipophilic moiety to a phosphorylated (deoxy)nucleoside analog will improve the activity of the drugs by circumventing the rate-limiting activation step of (deoxy)nucleoside analogs. Duplex and multiplex drugs consist of distinct active drugs with different mechanisms of action, which are linked to each other with either a lipid or a phosphodiester. Enzymatic cleavage of such a prodrug inside the cell releases the drug or the phosphorylated form of the drug. Antitumor activity of cytotoxic drugs can also be enhanced by the use of nanoparticles as carriers. Nanoparticles have the advantage of high stability, high carrier capacity, incorporation of hydrophobic and hydrophilic compounds and variable routes of administration. Encapsulating drugs in liposomes protects the drug against enzymatic breakdown in the plasma and makes it possible to get lipophilic compounds to the tumor site. Nanoparticles and liposomes can be used to target drugs either actively or passively to the tumor. In this review we discuss the considerable progress that has been made in increasing the efficacy of classic (deoxy)nucleoside and fluoropyrimidine compounds by chemical modifications and alternative delivery systems. We expect that combining different strategies could further increase the efficacy of these compounds. PMID- 19903146 TI - The Yin and Yang of CD4(+) regulatory T cells in autoimmunity and cancer. AB - The immune system balances effector responses with tolerance, to protect the host from pathogens while minimizing local damage to tissue. An altered control of immune homeostasis can lead to loss of tolerance to self antigens in autoimmunity, or promote unwanted tolerance to tumor growth. This review focuses on the dual activity of CD4(+) regulatory T cells (Tregs) in autoimmunity and cancer. Tregs play a key role in the mechanisms of immune tolerance and actively suppress pro-inflammatory responses, thus providing a beneficial action in autoimmunity and detrimental effects in cancer. PMID- 19903147 TI - Impairment of T cell immunity by the respiratory syncytial virus: targeting virulence mechanisms for therapy and prophylaxis. AB - Worldwide, Respiratory Syncytial Virus (RSV) causes severe bronchiolitis and pneumonia in children, the elderly and immuno-compromised individuals. Moreover, RSV is the mayor cause of infant hospitalization due to lower respiratory infection, regardless socioeconomic status. Accumulating data support the notion that immune responses elicited against naturally acquired RSV infections are non lasting and inappropriate for efficient virus clearance. Although there is consensus over the capacity of RSV to impair the development of an effective and protective adaptive immune response, very little is known about specific viral determinants involved in these processes as well as the molecular mechanisms developed by this virus to inhibit T cell function. Recent studies have provided evidence supporting an important role for dendritic cells in RSV-induced suppression of immunity. Although recognized for over 50 years as an important respiratory pathogen and healthcare problem, to date there are no available vaccines against this virus, which highlights the complexity of RSV-induced immunopathology. The development of new prophylactic and therapeutic tools against RSV requires the unveiling of molecular mechanisms and virulence factors responsible for the pathogenesis caused by this virus. In this review, we discuss recent findings describing virulence mechanisms evolved by RSV to negatively modulate the adaptive immune response in the host. Furthermore, novel strategies aimed to induce efficient T cell immunity against RSV are reviewed. PMID- 19903148 TI - Quantification of low-density and high-density lipoproteins in human serum by material enhanced infrared spectroscopy (MEIRS). AB - A key risk factor in the development of atherosclerosis is a high concentration of serum low density lipoprotein (LDL)-cholesterol. The main purpose of this study was to assess the LDL and high density lipoprotein (HDL) content in human serum by employing near-infrared (NIR) spectroscopy and multivariate calibration techniques. Initially a qualitative principal component analysis (PCA) based cluster model was generated to evaluate the feasibility of NIRS for classifying and identifying the LDL and HDL-cholesterol. Therefore TiO(2) beads were used as an adsorbent for selectively immobilizing LDL and HDL-cholesterol and further analysing the incubated and washed samples via NIR diffuse reflection spectroscopy. A principle component regression (PCR) model of 24 LDL standards in a range from 500 - 3000 ppm (clinical value is 1500 ppm) and a partial least squares regression (PLSR) model of 25 HDL standards in a range from 100 - 1000 ppm (clinical value is 400 ppm) were computed. Furthermore, the wavenumber region between 4000 cm(-1) and 7240 cm(-1) was found comprising the main spectral information regarding the TiO(2)-LDL and TiO(2)-HDL composites. The regression coefficients (r) for LDL and HDL were > 0.99 (calibration curve) and > 0.97 (validation curve), respectively. The PCR model of TiO(2)-LDL showed a standard error of estimation (SEE) of 122.80 ppm and a standard error of prediction (SEP) of 121.15 ppm while the PLSR model of TiO(2)-HDL showed 47.70 and 47.14 ppm, respectively. In order to determine the concentration of HDL in real serum samples, LDL was removed by adding a precipitation reagent containing 10 mg/mL magnesium dextran-sulfate, followed by incubation and centrifugation. The pretreated serum samples were predicted by the PLSR model while the standard deviation (SD) from the reference to the NIR predicted values of six test samples in a concentration range from 500 - 2500 ppm showed < 10 %. These results indicate the usefulness of the NIR spectroscopy (NIRS) as a potential alternative or even supplementary clinical method for the quick determination of LDL and HDL in human serum. PMID- 19903150 TI - Lymphatics and inflammation. AB - Inflammation is a local or systemic tissue reaction caused by external or internal stimuli with the objective to remove the noxa, inhibit its further dissemination and eventually repair damaged tissue. Blood vessels and perivascular connective tissue are important regulators of the inflammatory process. After a short initial ischemic phase, inflamed tissue is characterized by hyperaemia and increased permeability of capillaries. Therefore, blood vessels have been in the focus of inflammation research for quite some time, whereas lymphatic vessels have been neglected. Their reactivity is not immediately obvious, and, their identification within the tissue has hardly been possible until lymphatic endothelial cell (LEC)-specific molecules have been identified a few years ago. This has opened up the possibility to study lymphatics in normal and diseased tissues, and to isolate LECs for transcriptome and proteome analyses. Initial studies now provide evidence that lymphatics are not just a passive route for circulating lymphocytes, but seem to be directly involved in both the induction and the resolution of inflammation. This review provides a summary on the basics of inflammation, the structure of lymphatics and their molecular markers, human inflammation-associated diseases and their relation to lymphatics, animal models to study the interaction of lymphatics and inflammation, and finally inflammation-associated molecules expressed in LECs. The integration of lymphatics into inflammation research opens up an exciting new field with great clinical potential. PMID- 19903149 TI - Hypoxia inducible factor 1 as a therapeutic target in ischemic stroke. AB - In stroke research, a significant focus is to develop therapeutic strategies that prevent neuronal death and improve recovery. Yet, few successful therapeutic strategies have emerged. Hypoxia-inducible factor 1 (HIF-1) is a key regulator in hypoxia. It has been suggested to be an important player in neurological outcomes following ischemic stroke due to the functions of its downstream genes. These include genes that promote glucose metabolism, angiogenesis, erythropoiesis, and cell survival. Many lines of evidence have shown that HIF-1 is induced in ischemic brains. Importantly, it seems that HIF-1 is primarily induced in the salvageable tissue of an ischemic brain, penumbra. However, the effect of HIF-1 on neuronal tissue injuries is still debatable based on evidence from in vitro and preclinical studies. Furthermore, it is of importance to understand the mechanism of HIF-1 degradation after its induction in ischemic brain. This review provides a present understanding of the mechanism of HIF-1 induction in ischemic neurons and the potential effect of HIF-1 on ischemic brain tissue. The author also elaborates on potential therapeutic approaches through understanding of the induction mechanism and of the potential role of HIF-1 in ischemic stroke. PMID- 19903151 TI - The drug targeting and delivery approach applied to pt-antitumour complexes. A coordination point of view. AB - Platinum-based anticancer chemotherapy is associated to severe side effects because of its poor specificity. In particular, the hydrolysis of Pt-based drugs generates cationic complexes with electrophylic properties able to target DNA. The effectiveness of this kind of chemotherapy relies solely on the proliferation index of tumour cells, which is higher than in healthy cells. In recent years, the "drug targeting and delivery" approach has been developed in an attempt to reduce chemotherapy-related systemic side effects by using vectors that selectively deliver the cytotoxic agent to tumour cells, thus sparing healthy cells. These vectors include bioactive substances, such as nutrients, that more readily enter metabolically active tumour cells, or hormones, folates and bile acids, that are selectively conveyed by receptors/transporters often over expressed in cancer cells (active targeting). Alternatively, macromolecular vectors, exploiting the so-called EPR (enhanced permeability and retention) effect, can be used (passive targeting). The bioactive or macromolecular vector must contain a coordinating arm capable of binding the PtX(2)-unit, acting either as carrier or leaving group for the cytotoxic Pt-moiety. In both cases, the Pt vector conjugate should be promptly cleaved to generate the active species. The release of platinum drugs from the pharmacophore is crucial for fine-tuning of the overall cytotoxic properties of the conjugates. The "drug targeting and delivery" method represents an exciting field of research for improving the therapeutic potential of the long established, very efficient, but intrinsically non-specific Pt-based drugs. PMID- 19903152 TI - Expanding the chemical biologist's tool kit: chemical labelling strategies and its applications. AB - Methods that allow visualisation of proteins in living systems, in real time have been key to our understanding of the molecular underpinnings of life. Although the use of genetically encoded fusions to fluorescent proteins have greatly advanced such studies, the large size of these tags and their ability to perturb protein activity has been major limitations. Attempts to circumvent these issues have seen the genesis of complementary strategies to chemically label/modify proteins. Thus, chemical labelling approaches seek to "decorate" biomolecules in live cells through the site-specific introduction of a small, non-native chemical tag (or reporter group). The introduced tag is minimally invasive such that the activity and/or function of the target molecule in not perturbed/compromised by its inclusion. In most cases, this modification is brought about by fusing target biomolecules to protein domains/peptide tags or via the incorporation of reactive "handles" by either exploiting the cell's biosynthetic machinery or during protein synthesis. Selective tagging of the biomolecule then proceeds via a bioorthogonal chemical reaction following exogenous addition of probe(s). Depending on the nature of the probe, the method can be applied to either visualise/track the dynamics of target molecule(s) in their native cellular milieu or for affinity enrichment for further downstream applications. The versatility of these approaches has been demonstrated by their ability to tag not just proteins but also intractable biomolecules like lipids and glycans. In this review, we summarise the various strategies available to "chemically" tag proteins and provide a comparative analysis their advantages and disadvantages. We also highlight the many creative applications of such methodologies and discuss their future prospects. PMID- 19903153 TI - Dietary n-3 PUFA vascular targeting and the prevention of tumor growth and age related macular degeneration. AB - The protective role of dietary n-3 polyunsaturated fatty acids (PUFAs) against cardiovascular diseases has been partly related to their ability to modulate the risk condition known as "endothelial dysfunction", by reverting the endothelial alterations associated to it (reduced vascular reactivity, the proinflammatory state, and the prothrombotic properties). Moreover, vasculature represents the target for inhibition of pathologic neo-angiogenesis by n-3 PUFAs. This effect is believed to contribute to the beneficial action of these fatty acids against disorders which recognize neovascularization as a crucial pathogenetic step for their development, such as cancer and age-related macular degeneration (AMD). Many epidemiological studies have been conducted to evaluate the association between the intake of these fatty acids and the risk of developing cancer or AMD, even though contrasting and not definitive results have been obtained. Conversely, plenty of preclinical and in vitro experimental studies have provided evidence for the anti-angiogenic effects of n-3 PUFAs, mainly studying neo angiogenesis in general (using normal endothelial cells in vitro) or as a step of cancer growth. The main aim of this review is to critically review the current evidence for the inhibition of the neo-angiogenic process exerted by n-3 PUFAs in cancer and AMD, and to identify possible molecular mechanisms that might contribute to their beneficial effects. PMID- 19903154 TI - The protein therapy of kallikrein in cerebral ischemic reperfusion injury. AB - OBJECTIVE: Biomacromolecule like exogenous Kallikrein is difficult to pass through biomembrane and blood brain barrier. So, the use of exogenous Kallikrein for the therapy of nervous system diseases is restricted. We constructed the Protein Transduction Domain-Kallikrein (PTD-Kallikrein), checked its function of penetration and biotoxicity, and observed its influence on neurons and ischemic brain tissues. METHODS: PTD-Kallikrein (tissue kallikrein) was prepared by chemical synthesis. After PTD-Kallikrein injected 2.5 hours, rats brains were taken out and contents of Kallikrein were quantitated to observe the function of passing through blood brain barrier. Cell survival rate were measured by XTT methods to determine the peptide's biotoxicity. Apoptosis were inspected by TUNEL. PTD-Kallikrein was administrated immediately after cerebral ischemia. 24h later, infarct volume was determined by TTC stain and IL-1beta, TNF-alpha as well as PGE2 were measured by ELISA. RESULTS: 1. PTD-Kallikrein can pass through the biomembrane and blood brain barrier; 2. PTD-Kallikrein itself has no obviously biotoxicity. 3. PTD-Kallikrein increases cell survival rate, decreases neurons apoptosis during OGD/recovery; 4. HOE140 inhibits the effects of PTD-Kallikrein. 5. PTD-Kallikrein improves neurological impairment, decreases the infarct volume, and inhibits the release of IL-1beta, TNF-alpha, PGE2. 6. HOE140 inhibits the effects of PTD-Kallikrein on ischemia-reperfusion injury. CONCLUSIONS: 1. PTD Kallikrein can pass through the biomembrane and BBB efficiently and itself has no obviously biotoxicity. 2. PTD-Kallikrein has neuroprotective effect on neurons and cerebral ischemia injury. 3. PTD-Kallikrein is partially mediated by B2 receptors. PMID- 19903155 TI - Protein cysteine modifications: (2) reactivity specificity and topics of medicinal chemistry and protein engineering. AB - Cysteine (cysteinyl residue) modifications in proteins result in diversity in protein functions. The reaction specificity of a protein with a modified cysteine residue is determined by the overall conditions of the protein, including the spatial position of the cysteine residue, electrostatic interactions between cysteine residue and other charged residues, spatial interactions between the cysteine residue and a chemical compound, electrophilicity of the chemical compound, and the pH of the solution. In cysteine-dependant enzymes, each specific type of cysteine modification characterizes the catalytic mechanism of the enzyme. Recently, the catalytic mechanisms of peroxiredoxins and cysteine proteases, which contain a cysteine residue(s) in their catalytic sites, have been elucidated. In the catalytic process of peroxiredoxins, a sulfenyl intermediate is formed by oxidation of the catalytic cysteine residue. On the other hand, in cysteine proteases, the catalytic cysteine residue reacts with the carboxyl carbon of a peptide substrate to form an intermediate complex via S alkylation. In this review, we introduce the most current information on the applications of cysteine thiol chemistry for in vitro glycoprotein synthesis. Recently, a glycoprotein (monocyte chemotactic protein-3), containing an intact human complex-type sialyloligosaccharide has been chemically synthesized. The procedure used for this could have applications in the development of new protein based drugs, including antineoplastic drugs and antibiotics. It can also potentially be applied for improving the half-life and reducing the toxicity of these drugs, and for preventing the development of multidrug resistance. PMID- 19903156 TI - Synthesis and biological activity of Wuweizisu C and analogs. AB - Lignans are widely distributed in nature. The earliest recorded medicinal use of lignans dated back to over 1000 years ago. Lignan-rich plant products were also active ingredients in Chinese and Japanese folk medicines for the treatment of various diseases. The dried root and stem of this plant are listed in the Chinese pharmacopoeia for the treatment of rheumatoid arthritis, gastric, duodenal ulcers and many other diseases. This review highlights synthetic strategies for the Wuweizisu C analogs and the important pharmacological activities as well as therapeutic findings related to the treatment of HBV and other diseases. Notably a significant and ongoing project on Wuweizisu C and its analogs has led to the discovery and development of two potent derivatives alpha-DDB and BICYCLOL which are currently in clinical trials against HBV, especially in lowering elevated SGPT levels. Further design, synthesis, and evaluation of Wuweizisu C analogs are discussed. PMID- 19903157 TI - Nordihydroguaiaretic acid analogues: their chemical synthesis and biological activities. AB - Nordihydroguaiaretic acid is a natural occurring lignan mainly isolated and commercially produced from desert plant, creosote bush (Larrea divaricata Cav. Or Corillea tridentate), which can be widely found in the border zone of southern of USA and northern of Mexico. During past 100 years, extensive research has demonstrated that nordihydroguaiaretic acid and its synthetic analogues are potentially useful in treating diseases related to cancers, diabetes, viral, bacterial infections, and inflammation. Remarkably, terameprocol, a tetra-O methyl derivative of nordihydroguaiaretic acid, is currently in Phase I/II clinical trials as an anticancer agent. This review deals with the chemical synthesis and bioactivities of nordihydroguaiaretic acid and its structurally related derivatives, which possess anticancer, antioxidant, antimicrobial, anti inflammatory and immunosuppressive activities. The review consists of the data reported in over 100 publications. PMID- 19903158 TI - Chemo-enzymatic transformation of taxanes and their reversal activity towards MDR tumor cells. AB - Over 200 derivatives have been obtained through chemo-enzymatic transformation of the taxanes derived from cell cultures of Taxus chinensis. The reversal activity towards MDR tumor cells and cyto-toxicity of most these compounds were evaluated, and several derivatives exhibited powerful MDR reversal activity. The substrate specificities of the two most important biotransformation reactions-C-7beta and 9alpha hydroxylations, were investigated and preliminarily concluded. In addition, some key intermediates in hypothetically biosynthetic pathway of taxoid were obtained. These results indicate that biotransformation combined with chemical transformation is a powerful approach to structural diversification of natural products, lead compound discovery and the study on the structure activity relationships of the derivatives. Moreover, biotransformation may mimic some steps in biosynthetic pathway of natural products and provide useful hints as to their biogenetic study. PMID- 19903159 TI - The potential of natural products as effective treatments for allergic inflammation: implications for allergic rhinitis. AB - The impact of natural products on human health has been enormous, and the study of natural products continues to influence research in the fields of chemistry, biology, and ecology. Historically, the majority of our medicines originate from natural products and their synthetic derivatives, many of which have taught us valuable lessons about biology. While advances in synthetic and combinatorial chemistry have given rise to notable successes in the development of new drugs, the perceived value of natural products in the treatment of allergic disease has yet to be fully explored. The immune system is a highly complex, intricately regulated group of cells whose integrated function is essential to health. Cells of the immune system may interact in a cell-cell manner and may also respond to intercellular messages including hormones, cytokines, and effector molecules produced by various cells. These effector molecules include histamine, kinins, leukotrienes, prostaglandins, and serotonin. The immune system can be modified by diet, pharmacologic agents, environmental pollutants, and naturally occurring food chemicals, such as vitamins and flavonoids. Allergic inflammation is mediated by several types of immune cells all of which can be effected by these naturally occurring bioactive compounds but this review will focus on mast cells and their mediators since these cells are the focal point of allergic reactions such as allergic rhinitis. The molecular mechanisms and scientific validity of some herbal remedies currently used clinically in the treatment of allergic rhinitis will be explored. PMID- 19903160 TI - Biosynthesis of bacterial aromatic polyketides. AB - Aromatic polyketides represent important members of the family of polyketides, which have displayed a wide assortment of bioactive properties, such as antibacterial, antitumor, and antiviral activities. Bacterial aromatic polyketides are mainly synthesized by type II polyketide synthases (PKSs). Whereas malonyl-CoA is exclusively used as the extender unit, starter units can vary in different aromatic polyketide biosynthetic pathways, leading to a variety of polyketide backbones. Once the polyketide chains are elongated by the minimal PKSs to the full length, the immediate tailoring enzymes including ketoreductases, oxygenases and cyclases will work on the nascent chains to form aromatic structures, which will be further decorated by those late tailoring enzymes such as methyltransferases and glycosyltransferases. The mechanistic studies on the biosynthetic pathways of aromatic polyketides such as oxytetracycline and pradimicin A have been extensively carried out in recent years. Engineered biosynthesis of novel "unnatural" polyketides has been achieved in heterologous hosts such as Streptomyces coelicolor and Escherichia coli. This review covers the most recent advances in aromatic polyketide biosynthesis, which provide new enzymes or methods for building novel polyketide biosynthetic machinery. PMID- 19903161 TI - Pentacyclic triterpenoids and their saponins with apoptosis-inducing activity. AB - Pentacyclic triterpenoids exert their antitumor activity through different mechanisms, one of which is apoptosis induction. Although there are many reports on the apoptosis inducing activity of pentacyclic triterpenoids, a systematic survey and discussion on their structure-activity relationships (SARs) is still lacking. In this review, we summarized such activity of oleanane, ursane and lupane type triterpenoids, the most abundant pentacyclic triterpene prototypes in plants. Their structural characteristics responsible for the activity are also discussed, in order to unravel their SARs for the apoptotic induction and benefit further investigations on the antitumor triterpenoids. PMID- 19903162 TI - Polyisoprenylated benzophenones from Clusiaceae: potential drugs and lead compounds. AB - Many new polyisoprenylated benzophenones with a bicyclo[3.3.1]-nonane-2,4,9 trione core structure have been isolated from plants in the Clusiaceae family, and their potent biological properties have been the subject of several studies. This review summarizes the biological activities reported for these secondary metabolites including cytotoxic, antimicrobial, antioxidant, and anti inflammatory activities. Our efforts during the past years have foremost been directed towards isolating new polyisoprenylated benzophenones, as well as understanding the possible target and mechanism of action through which these compounds arrest cancer cells and inhibit the progression of the cell-cycle. The transcription of genes is affected in cancer cells treated with polyisoprenylated benzophenones; the oncogene c-Myb is down-regulated and endoplasmatic stress genes XBP1, ATF4, and DDIT3/CHOP are turned on. Consequently, the expression of iNOS and cell cycle regulators such as cyclin D and E are reduced. Evidence presented by independent investigators suggests that polyisoprenylated benzophenones affect the mediators in the Akt/mTOR stress pathway, although the specific target remains to be discovered. In addition, benzophenones isolated from plants display high antioxidant capacity and protect cells from oxidative stress and the formation of ROS involved during the inflammatory process. Since antiviral activity was initially reported for guttiferone A, potent synthetic analogues have been developed as effective new non-nucleoside reverse transcriptase inhibitors (NNRTI) to treat drug resistant HIV-1. In addition, benzophenones exert antimicrobial effects particularly against MRSA. The structure-activity relationships of polyisoprenylated benzophenones from natural sources and those of synthetic analogues are included in this review. Absorption, metabolism, and elimination of benzophenones are also discussed. PMID- 19903163 TI - Low molecular weight and oligomeric chitosans and their bioactivities. AB - Chitosan is one of the most abundant marine-based biopolymers. Low molecular weight and oligomeric chitosans are water-soluble hydrolysates of chitosan. They have been shown to have a wide range of biological activities and industrial applications. In particular, low molecular weight and oligomeric chitosans have been reported to have the health benefits such as immunity regulation, anti tumor, liver protection, blood lipids lowering, anti-diabetic, antioxidant and anti-obesity. In this paper, the preparation and analytical methods, and bioactivities of these low molecular weight and oligomeric chitosans were reviewed, with the latest progresses introduced and discussed. PMID- 19903164 TI - Anticancer drugs discovery and development from marine organism. AB - The chemical and biological diversity of the different marine evolutionary group is endless and therefore, this is an amazing resource for the discovery of new anticancer drugs. Comprising 34 of the 36 Phyla of life, marine ecosystems are indeed our last genetic diversity and biotechnological boundary; terrestrial systems possess only 17 Phyla. Sponges, coelenterates and microorganisms are the foremost resources of therapeutic compounds. Algae, echinoderms, tunicates, mollusks, bryozoans are also the sources of anticancer drugs from marine resources. We highlight the past and current status of marine anticancer pharmacology using different marine groups. PMID- 19903165 TI - Exploring and exploiting microbial diversity through metagenomics for natural product drug discovery. AB - Microorganisms of millions species exist in every corner of the Earth, and form a dynamic genetic reservoir that are not clearly revealed and categorized due to barrier in current cultivation technology. Their applications in biomedical and environmental aspects are more than satisfactory. However, the situation has drastically changed during the turn of the century because of the rapid development of phylogenetic studies based on rRNA sequencing independent of standard laboratory cultivation. More recently, high throughput sequencing technology which enables direct sequencing of community DNA for metagenomic analyses are making a direct impact on our understanding of microbial diversity, ecology, and secondary metabolism. In this review, we highlight some recent progress and innovation on metagenomic research with an emphasis on natural product drug discovery. The rapid path of accumulating decoded metagenomics would be an efficient guide on direct access to the genomes of numerous non-culturable microorganisms for their genomic diversity and associated chemical prosperity for potential medicinal applications. PMID- 19903166 TI - Chemical and biological explorations of the family of CC-1065 and the duocarmycin natural products. AB - CC-1065, the duocarmycins and yatakemycin are members of a family of ultrapotent antitumour antibiotics that have been the subject of extensive investigations due to their mode of action and potential in the design of new anticancer therapeutics. The natural products and their analogues exert their effects through a sequence selective alkylation of duplex DNA in the minor groove at the N3 of adenine. An understanding of their structure and its effect on biological activity has been derived through chemical synthesis and has also generated new potential lead compounds. These studies form the first section of the review. The desire to progress these compounds to clinic has also led to studies of bioconjugation and prodrug formation and this is discussed in the second section of the review. The combination of synthesis with key biological experiments is a powerful tool to define the requirements for the development of natural products as potential therapeutic agents. The studies described herein form an excellent paradigm for the study and development of other natural products. PMID- 19903167 TI - Bivalirudin: alternative anticoagulation during cardiopulmonary bypass in patients with heparin-induced thrombocytopenia. AB - An estimated 1-3% of patients who receive therapeutic anticoagulation with unfractionated heparin (UFH) develop antibodies to heparin with concomitant development of thrombocytopenia, defined as HIT or Heparin-Induced Thrombocytopenia. HIT complicates the management of patients presenting for cardiac surgery, particularly those who need cardiopulmonary bypass (CPB) which requires a large dose of UFH. A portion of these patients will have significant thrombotic complications referred to as HITT (Heparin-induced thrombocytopenia with thrombosis). In patients with established or suspected HIT, all heparin must be withheld and an alternative form of anticoagulation utilized for CPB. Various approaches and pharmacological alternatives have been described but no regimen has replaced the routine use of UFH anticoagulation with protamine reversal after CPB. We review the use of bivalirudin as a reliable and safe alternative anticoagulation strategy during cardiopulmonary bypass with specific emphasis on patients with HIT and outlining some recent patents. PMID- 19903168 TI - A novel harvesting method for cultured cells using iron-cross-linked alginate films as culture substrates. AB - The present study was conducted to assess the efficiency of a novel cell harvesting method involving dissolution of the culture substrate composed of Fe alginate (ferric-ion-cross-linked alginate). Cell harvesting is an essential step for recovery of cultured adherent cells, but conventional methods such as trypsinization or scraping cause considerable damage to the cells. We therefore devised an original method for harvesting cultured cells using Fe-alginate films as a culture substrate and then retrieving the cells by disintegration of the alginate gel. Fe-alginate was easily dissolved under physiological conditions by exchange of cross-linked ions using chelating agents such as citrate. The effects of this cell-harvesting method were investigated in comparison with trypsinization and scraping. Cells harvested by dissolution of the Fe-alginate substrate showed high viability and metabolic activity, high recovery rate on subculture, a low degree of membrane damage with superior expression of cell adhesion molecules (beta1 integrin and N-cadherin), high resistance to oxidative stress, and high viability, metabolic activity and recovery rate after cryopreservation, in contrast with the cells harvested by trypsinization or scraping from tissue-culture polystyrene substrates. These results suggest that our new system involving dissolution of Fe-alginate culture substrate is effective for cell harvesting and may be advantageous for biomedical applications. PMID- 19903169 TI - Activation of autophagy through modulation of 5'-AMP-activated protein kinase protects pancreatic beta-cells from high glucose. AB - Chronic hyperglycaemia is detrimental to pancreatic beta-cells by causing impaired insulin secretion and diminished beta-cell function through glucotoxicity. Understanding the mechanisms underlying beta-cell survival is crucial for the prevention of beta-cell failure associated with glucotoxicity. Autophagy is a dynamic lysosomal degradation process that protects organisms against metabolic stress. To date, little is known about the physiological function of autophagy in the pathogenesis of diabetes. In the present study, we explored the roles of autophagy in the survival of pancreatic beta-cells exposed to high glucose using pharmacological and genetic manipulation of autophagy. We demonstrated that chronic high glucose increases autophagy in rat INS-1 (832/13) cells and pancreatic islets, and that this increase is enhanced by inhibition of 5'-AMP-activated protein kinase. Our results also indicate that stimulation of autophagy rescues pancreatic beta-cells from high-glucose-induced cell death and inhibition of autophagy augments caspase-3 activation, suggesting that autophagy plays a protective role in the survival of pancreatic beta-cells. Greater knowledge of the molecular mechanisms linking autophagy and beta-cell survival may unveil novel therapeutic targets needed to preserve beta-cell function. PMID- 19903170 TI - Robot-assisted radical cystectomy: intermediate survival results at a mean follow up of 25 months. AB - OBJECTIVE: To assess the overall and disease-specific survival rates of patients undergoing robot-assisted radical cystectomy (RARC) compared with historical open cystectomy. PATIENTS AND METHODS: Survival, pathological and demographic data were collected on all patients undergoing RARC for bladder cancer from both Tulane University Medical Center and Mayo Clinic Arizona. Of a total of 80 RARCs we only included those with a follow-up of > or =6 months from surgery. Survival curves were compared with those from historical series of open cystectomy. RESULTS: Of the 80 patients 59 were identified as having a follow-up of > or =6 months from the date of surgery. The mean (range) follow-up was 25 (6-49) months. Overall survival rates at 12 and 36 months were 82% and 69%, respectively, and disease-specific survival rates were 82% and 72% at 12 and 36 months, respectively. These results are comparable to survival rates from open cystectomy. As expected, patients with lymph node-positive disease fared worse than those with lymph node-negative disease. Patients with extravesical lymph node-negative disease (pT3, pT4) fared worse than patients with organ-confined lymph node-negative disease. Also, patients with lymph node-positive disease fared worse than those with extravesical lymph node-negative disease, which is consistent with historical results of open cystectomy. CONCLUSIONS: RARC has a comparable survival rate to open cystectomy in the intermediate follow-up. Further study with a longer follow-up and more patients is necessary to determine any long-term survival benefits. PMID- 19903172 TI - Clinical investigation of the novel iron-chelating agent, CP94, to enhance topical photodynamic therapy of nodular basal cell carcinoma: further explanation of a dose-escalating pilot study conducted primarily to consider the safety of this pharmacological modification. PMID- 19903171 TI - Characterization of R132H mutation-specific IDH1 antibody binding in brain tumors. AB - Heterozygous point mutations of isocitrate dehydrogenase (IDH)1 codon 132 are frequent in grade II and III gliomas. Recently, we reported an antibody specific for the IDH1R132H mutation. Here we investigate the capability of this antibody to differentiate wild type and mutated IDH1 protein in central nervous system (CNS) tumors by Western blot and immunohistochemistry. Results of protein analysis are correlated to sequencing data. In Western blot, anti-IDH1R132H mouse monoclonal antibody mIDH1R132H detected a specific band only in mutated tumors. Immunohistochemistry of 345 primary brain tumors demonstrated a strong cytoplasmic and weaker nuclear staining in 122 cases. Correlation with direct sequencing of 186 cases resulted in consensus of 177 cases. Genetic retesting of cases with conflicting findings resulted in a match of 186/186 cases, with all discrepancies resolving in favor of immunohistochemistry. Intriguing is the ability of mIDH1R132H to detect single infiltrating tumor cells. The very high frequency and the distribution of this mutation among specific brain tumor entities allow the highly sensitive and specific discrimination of various tumors by immunohistochemistry, such as anaplastic astrocytoma from primary glioblastoma or diffuse astrocytoma World Health Organization (WHO) grade II from pilocytic astrocytoma or ependymoma. Noteworthy is the discrimination of the infiltrating edge of tumors with IDH1 mutation from reactive gliosis. PMID- 19903173 TI - Cutaneous T-cell lymphoma (mycosis fungoides) relapsed with different immunological phenotype after bone marrow transplant. PMID- 19903174 TI - Methotrexate: a useful steroid-sparing agent in recalcitrant chronic urticaria. AB - Background Reports of methotrexate for chronic urticaria are anecdotal. Objectives To assess the effectiveness of methotrexate in steroid-dependent chronic urticaria, its impact on steroid reduction and any differences in response between patients with and without functional autoantibodies. Methods A retrospective case-note review of 16 patients with steroid-dependent chronic urticaria treated with methotrexate was carried out. Ten patients had chronic ordinary/spontaneous urticaria (CU), including three with associated delayed pressure urticaria; four patients had normocomplementaemic urticarial vasculitis (UV); and two patients had idiopathic angio-oedema without weals. Median disease duration before methotrexate was 48.5 months (range 12-164). All were unresponsive to antihistamines and second-line agents, except prednisolone. Eleven were assessed for autoimmune urticaria with the basophil histamine release assay (n = 5), autologous serum skin test (n = 5) or both (n = 1). Response to methotrexate was scored: no benefit; some benefit (fewer weals and symptomatic improvement but no steroid reduction); considerable benefit (improvement with steroid reduction); or clear (no symptoms, off steroids but on antihistamines). Results Twelve of 16 patients (eight CU, three UV, one idiopathic angio-oedema) responded. Three showed some benefit, seven considerable benefit and two cleared. Four of eight responders and three out of three nonresponders showed evidence of functional autoantibodies. The dose to achieve a steroid-sparing effect was 10-15 mg weekly (cumulative dose range 15-600 mg, median 135 mg). Methotrexate was well tolerated. Conclusions Methotrexate may be a useful treatment for steroid dependent chronic urticaria. Functional autoantibodies do not correlate with response. The beneficial effects of methotrexate may be anti-inflammatory and immunosuppressive. It may therefore benefit chronic urticaria independently of the pathogenic mechanism, whether autoimmune or not. PMID- 19903175 TI - Investigation of papulopustular eruptions caused by cetuximab treatment shows altered differentiation markers and increases in inflammatory cytokines. AB - BACKGROUND: Epidermal growth factor receptor (EGFR) critically regulates tumour cell division, survival and metastasis. Agents that inhibit EGFR have been used in the treatment of advanced-stage malignancies, but cause variable cutaneous side-effects, most often papulopustular eruptions and xerosis. OBJECTIVES: We assayed expression of inflammatory cytokines [interleukin (IL)-1alpha, tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, human leucocyte antigen (HLA)-DR and intercellular adhesion molecule (ICAM)-1], differentiation markers (filaggrin, involucrin and loricrin) and phosphorylated EGFRs (pEGFRs) in papulopustular eruptions to determine the association between these markers and the eruptions caused by cetuximab. PATIENTS/METHODS: Twelve papulopustular lesion biopsies were selected from patients with colon cancer who had received cetuximab treatment. Immunohistochemistry and immunofluorescence with a confocal laser scanning microscopy were performed. RESULTS: Filaggrin expression decreased and expression of involucrin, various inflammatory markers (IL-1alpha, TNF-alpha, ICAM-1 and HLA-DR) increased and the expression of pEGFR was markedly downregulated in papulopustular eruptions. In perilesions, decreased pEGFR expression was noted in hair follicles compared with interfollicular epidermis. The increase of IL-1alpha and TNF-alpha was observed in perilesions as in the lesions. CONCLUSIONS: The early inflammatory events (IL-1alpha and TNF-alpha expression) seen, and the lack of pEGFR in perilesional follicles, indicate that inflammatory events induced by EGFR inhibition may initiate papulopustular eruptions along with the altered differentiations. The decrease of filaggrin may contribute to the pathogenesis of the xerosis caused by cetuximab. PMID- 19903176 TI - Increased expression of glucocorticoid receptor beta in lymphocytes of patients with severe atopic dermatitis unresponsive to topical corticosteroid. AB - BACKGROUND: Variable response to topical glucocorticoid therapy occurs in the treatment of severe atopic dermatitis (AD). Glucocorticoid receptor (GR)-beta does not bind glucocorticoids but antagonizes the activity of the classic GRalpha, and could thus account for glucocorticoid insensitivity. OBJECTIVES: To investigate GRalpha and GRbeta mRNA and protein expression in lymphocytes of patients with AD before and after treatment with topical corticosteroids. METHODS: Blood was collected from 11 healthy donors, 10 patients with mild AD and 13 patients with severe AD. mRNA was isolated from peripheral blood mononuclear cells. Expression of GRalpha and GRbeta mRNA was determined by reverse transcriptase-polymerase chain reaction and quantitated. Expression of the GRs was confirmed by Western blot analysis. RESULT: The expression of GRalpha mRNA was detected in all subjects. GRbeta mRNA was detected in four out of 11 healthy volunteers, five out of 10 patients with mild AD and 11 out of 13 patients with severe AD. The incidence of GRbeta mRNA expression was higher in patients with severe AD (85%) than in patients with mild AD (50%), and significantly higher than in healthy volunteers (36%, P = 0.033). Four of the 13 patients with severe AD showed a 3.3-13.2-fold increase in the expression of GRbeta mRNA during a 2 week treatment with topical corticosteroids. In these patients the response to topical corticosteroids was poor. CONCLUSIONS: Expression of GRbeta is increased during topical corticosteroid treatment in the lymphocytes of patients with AD and, in particular, glucocorticoid-insensitive AD is associated with increased expression of GRbeta. PMID- 19903177 TI - Narrowband ultraviolet B phototherapy in the treatment of cutaneous graft-versus host disease in oncohaematological paediatric patients. AB - BACKGROUND: Graft-versus-host disease (GVHD) represents an important complication following allogeneic bone marrow transplantation. In recent years, narrowband ultraviolet B (NB-UVB, 311-313 nm) has been found to be a beneficial adjuvant treatment in patients refractory to first-line immunosuppressive drugs. OBJECTIVES: The aim of this study is to analyse retrospectively the clinical outcome of 10 GVHD paediatric patients treated with NB-UVB therapy. PATIENTS AND METHODS: Ten paediatric patients (six girls and four boys: median age 12.5 years, range 4-20) with cutaneous GVHD were enrolled in the study: five patients with chronic GVHD and five patients with an overlap syndrome GVHD. All patients had already been shown to be resistant to first-choice immunosuppressive protocols, and were treated with NB-UVB phototherapy until a clinical remission of skin lesions occurred. RESULTS: A complete response (absence of lesions) was achieved in 80% of the cases (eight patients) after a median number of 29 treatments, corresponding to a median of 7.5 weeks (52 days) of treatment (range 3-13 weeks), with an average cumulative dose of 28.71 J cm(-2) (range 1.02-70.38 J cm(-2)). Only two patients reported a partial remission (< 18% of body surface area involved). During the follow-up period, a complete remission after 1 year was observed in 75% of patients and after 2 years in 71% of the evaluable patients. CONCLUSIONS: This study provides evidence that NB-UVB phototherapy represents a valid second-line treatment in paediatric patients affected by GVHD and refractory to immunosuppressive first-line treatment. PMID- 19903178 TI - Complex multipathways alterations and oxidative stress are associated with Hailey Hailey disease. AB - BACKGROUND: Hailey-Hailey disease (HHD) is an autosomal dominant disorder characterized by suprabasal cutaneous cell separation (acantholysis) leading to the development of erosive and oozing skin lesions. While a strong relationship exists between mutations in the gene that encodes the Ca(2+)/Mn(2+)-adenosine triphosphatase ATP2C1 and HHD, we still have little understanding of how these mutations affect manifestations of the disease. OBJECTIVES: This study was designed to determine early signalling events that affect epithelial cell growth and differentiation during HHD development. METHODS: Expression of key regulatory signals important for maintaining skin homeostasis were evaluated by Western blot analysis and by reverse transcriptase-polymerase chain reaction in primary keratinocytes obtained from skin biopsies of patients with HHD. Reactive oxygen species accumulation in primary keratinocytes derived from lesional skin of patients with HHD was assessed by dihydrorhodamine 123 (DHR) assay. RESULTS: HHD derived keratinocytes showed downregulation of both Notch1 and differential regulation of different p63 isoforms. Itch and p63 are co-expressed in the epidermis and in primary keratinocytes where Itch controls the p63 protein steady state level. We found that the Itch protein was significantly decreased in HHD derived keratinocytes whereas the expression of its target, c-Jun, remained unaffected. We also found that HHD-derived keratinocytes undergo oxidative stress, which may explain both Notch1 and Itch downregulation. CONCLUSIONS: Our attempt to explore the molecular mechanism underlying HHD indicates a complex puzzle in which multi-hit combinations of altered signal pathways may explain the wide spectrum of defects in HHD. PMID- 19903179 TI - Daily practice assessment of liver injury in patients with psoriasis on methotrexate. PMID- 19903180 TI - Felodipine-induced eruptive telangiectasia following mastectomy and radiotherapy. PMID- 19903181 TI - Molecular basis of EEC (ectrodactyly, ectodermal dysplasia, clefting) syndrome: five new mutations in the DNA-binding domain of the TP63 gene and genotype phenotype correlation. AB - Summary EEC (ectrodactyly, ectodermal dysplasia, clefting; OMIM 604292) syndrome is an autosomal dominant developmental disorder. Characteristic clinical features comprise abnormalities in several ectodermal structures including skin, hair, teeth, nails and sweat glands as well as orofacial clefting and limb defects. Pathogenic mutations in the TP63 transcription factor have been identified as the molecular basis of EEC syndrome and to date 34 mutations have been reported. The majority of mutations involve heterozygous missense mutations in the DNA-binding domain of TP63, a region critical for direct interactions with DNA target sequences. In this report, we present an overview of EEC syndrome, discuss the role of TP63 in embryonic development and skin homeostasis, and report five new TP63 gene mutations. We highlight the significant intra- and interfamilial phenotypic variability in affected individuals and outline the emerging paradigm for genotype-phenotype correlation in this inherited ectodermal dysplasia syndrome. PMID- 19903182 TI - Merocel nasal packing with airway: a method to help secure full-thickness skin grafts on the nasal tip. PMID- 19903183 TI - Ustekinumab improves health-related quality of life in patients with moderate-to severe psoriasis: results from the PHOENIX 1 trial. AB - Background PHOENIX 1 was a phase III, randomized, double-blind, placebo controlled study that demonstrated the long-term efficacy and safety of ustekinumab in patients with moderate-to-severe psoriasis. Objectives To assess the effect of ustekinumab maintenance therapy on health-related quality of life (HRQoL) in PHOENIX 1 patients. Patients and methods Patients (n = 766) were randomized to receive ustekinumab 45 mg (n = 255) or 90 mg (n = 256) at weeks 0 and 4 and every 12 weeks thereafter, or placebo (n = 255) at weeks 0 and 4 with crossover to ustekinumab at week 12. Ustekinumab-randomized patients achieving at least 75% improvement in Psoriasis Area and Severity Index (PASI) 75 at weeks 28 and 40 were re-randomized at week 40 to continue ustekinumab or be withdrawn until loss of therapeutic effect. HRQoL was assessed using the SF-36 and Dermatology Life Quality Index (DLQI). Results At baseline, more than 97% had a DLQI > 1 and the average DLQI was > 10, indicating a significant impact on patients' HRQoL. Significantly greater proportions of patients receiving ustekinumab 45 and 90 mg achieved a normalized DLQI score (< or = 1) compared with placebo (53.2%, 52.4% and 6.0%, respectively, both P < 0.001) at week 12 and achieved a clinically meaningful improvement (increase of at least five points) in SF-36 physical (23.1%, 33.7% and 15.6%) and mental (25.5%, 31.3% and 14.8%) component summary scores. At week 12, changes in individual DLQI and SF-36 domains were significantly better in each ustekinumab group vs. placebo (P < 0.001). The magnitude of improvement across SF-36 scales was greatest for the bodily pain and social functioning domains. Improvements in HRQoL were sustained with maintenance ustekinumab therapy through at least 1 year. Regression analysis showed that, after adjustment for improvement in PASI or Physician's Global Assessment (PGA), ustekinumab-treated patients demonstrated significant improvements in DLQI. Conclusions Ustekinumab improves HRQoL in patients with moderate-to-severe psoriasis. Patient-reported outcomes measured a treatment effect beyond that indicated by clinical measures. PMID- 19903184 TI - Stroke prevention versus bleeding risk of vitamin-K antagonists: a double-edged sword in patients with atrial fibrillation who require surgery. AB - Patients with atrial fibrillation taking vitamin-K antagonists and undergoing invasive interventions or large surgery procedures are at highest risk of bleeding complications. Therefore, the temporary interruption of vitamin-K antagonists and bridging with heparin is a frequent clinical need, particularly in patients with high risk for stroke. The management of such patients is challenging because of the lack of randomized clinical trials assessing different periprocedural anticoagulation approaches and inconsistent recommendations from consensus groups. Recent non-randomized trials have helped to estimate the risks of thromboembolism and bleeding with "bridging" anticoagulation involving either low-molecular-weight heparin or intravenous unfractioned heparin. Nevertheless, there is still a clear need for randomized double-blinded controlled trials comparing efficacy and safety of the different "bridging" strategies, including unfractionated heparin and placebo comparators, in preventing thromboembolism for specific patients and procedures. PMID- 19903185 TI - Drug-drug noninteractions. AB - Understanding and documentation of drug-drug interactions (DDIs) are an important component of drug development, and of clinical therapeutics. Because clinical DDI studies are costly, time-consuming, and involve some risk, not all clinical DDI questions can be realistically addressed through human DDI trials. In vitro models have been used to identify and predict drug combinations that might interact, and combinations that are unlikely to interact. This screening or "filtration" information allows clinical resources to be targeted in a more informed way. Still, many DDI studies will end up with a negative result. Negative DDI results constitute important and clinically relevant information, and scientific reports of such studies are candidates for publication. PMID- 19903186 TI - Bridging of chronic oral anticoagulation with enoxaparin in patients with atrial fibrillation: results from the prospective BRAVE registry. AB - Current American College of Chest Physicians (ACCP) guidelines on the perioperative management of oral anticoagulation (OAC) suggest bridging therapy with therapeutic doses of low-molecular-weight heparin (LMWH) in patients with atrial fibrillation (AF) if at high or moderate thromboembolic (TE) risk, and with reduced doses in patients with low TE risk. Our objective was to assess the efficacy and safety of bridging OAC with enoxaparin in AF patients. These are the results of an open, prospective monocenter register. Hospitalized and ambulatory patients with AF requiring bridging therapy at high or moderate TE risk and normal renal function were treated with therapeutic LMWH doses; all other patients received reduced doses. A total of 703 patients were enrolled, of whom 358 (50.9%) were at moderate-to-high and 345 (49.1%) at low TE risk. Renal impairment was detected in 308 patients (43.8%). One hundred ninety patients (27.1%) were treated with therapeutic LMWH doses and 513 (72.9%) with reduced doses. No TE events were observed during the follow-up period (0%; 95% confidence interval [CI] 0.0-0.52). Three major bleeds (0.4%; 0.1-1.2) and 60 minor bleeds were noted (8.9%; 6.6-10.9). Age and total LMWH doses were risk factors for bleeding in the multivariate analysis. The study, under conditions of everyday clinical care, supports a predefined bridging regimen based on the individual patient's TE risk and renal function. Patients with low TE risk or with impaired renal function can be bridged effectively and safely with reduced LMWH doses. PMID- 19903188 TI - Gender and responses to aspirin and clopidogrel: insights using short thrombelastography. AB - There is significant variability in both baseline clotting tendency and response to antiplatelet therapy. Responses are associated with outcome. We have investigated whether differences could explain the increased risk observed in women presenting with coronary artery disease. We have utilized short thrombelastography to assess (i) baseline clotting responses, (ii) response to aspirin and clopidogrel, and (iii) post-treatment platelet reactivity in 48 young volunteers, 22 older patients and 18 patients with previous stent thrombosis. Baseline responses were significantly higher in young women than in men. While there was no difference in response to aspirin, platelet reactivity on aspirin remained higher in women (area under curve at 15 min [AUC15] of arachidonic acid channel 332 +/- 122 vs. 172 +/- 80, P= 0.04). Young women had less response to clopidogrel (% reduction in AUC15 with adenosine diphosphate [ADP] 36.4 +/- 12.4 vs. 64.0 +/- 13.2, P < 0.01) in addition to higher post-treatment reactivity (AUC15 of ADP 714 +/- 161 vs. 311 +/- 146, P < 0.01) compared to men. There were no such differences between male and female patients over 50. However, young women with previous stent thrombosis had among the highest platelet reactivity observed. Compared to men, young women have greater baseline clotting tendency, reduced response to clopidogrel, and greater post-treatment reactivity while on both aspirin and clopidogrel. Differences in clotting tendency and response to antiplatelet therapy may contribute to the excess risk observed in young women but are not observed in older female patients. PMID- 19903187 TI - Effects of laropiprant, a selective prostaglandin D(2) receptor 1 antagonist, on the pharmacokinetics of rosiglitazone. AB - Laropiprant (LRPT), a prostaglandin D(2) receptor-1 antagonist shown to reduce niacin-induced flushing symptoms, has been combined with niacin for treatment of dyslipidemia. This open-label, randomized, 2-period crossover study assessed the pharmacokinetics of single-dose rosiglitazone in the presence and absence of multiple-dose LRPT. Twelve healthy male and female subjects, 34-64 years of age, received two, once-daily oral treatments in random sequence separated by >/=3-day washout: (1) multiple-dose LRPT 40 mg/day for 7 days (Days 1 to 7) coadministered with single-dose rosiglitazone 4 mg on Day 6; (2) single-dose rosiglitazone 4 mg on Day 1. Comparability was declared because the 90% confidence interval (CI) for the AUC(0-infinity) geometric mean ratio (GMR; rosiglitazone + LRPT/rosiglitazone alone) [0.92 (0.86, 0.99)], was contained within prespecified bounds (0.70, 1.43). The C(max) GMR (90% CI) for rosiglitazone was 0.98 (0.95, 1.02). There was no evidence of clinically meaningful alterations in the pharmacokinetics of rosiglitazone, a probe CYP2C8 substrate, following coadministration of multiple dose LRPT in healthy subjects. Therefore, findings suggest that LRPT does not inhibit CYP2C8-mediated metabolism. PMID- 19903189 TI - Ability of amiodarone and propranolol alone or in combination to prevent post coronary bypass atrial fibrillation. AB - Atrial fibrillation (AF) is the most common arrhythmia in coronary artery bypass grafting (CABG) patients. The purpose of this study was to determine the best prophylaxis for AF prior to CABG. In this double-blind randomized study, 240 consecutive patients underwent elective CABG. They were then divided randomly into three groups to receive propranolol (n = 80), amiodarone (n = 80), or both drugs (n = 80). All groups received their medications from preoperative day 7 to post-CABG day 5. The patients were well matched for age, sex, risk factors, comorbidities, ejection fraction, and cardioplegic technique. Post-CABG AF developed in 22 patients (9.2%) of whom 13 (16.3%) had received propranolol, 5 (6.3%) had received amiodarone, and 4 (5%) had received both drugs. The difference between the propranolol group and the other two groups was statistically significant (P= 0.02), but that between the amiodarone and amiodarone + propranolol group was not significant. Age was a significant predictor of post-CABG AF (P= 0.034). Other factors such as diabetes, sex, hyperlipidemia, smoking, hypertension, family history, cerebrovascular accidents, left atrial size, and ejection fraction were not significant predictors of post CABG AF. Preoperative amiodarone or amiodarone with propranolol were more effective than propranolol in reducing the frequency of AF. There was a strong relationship between age and the development of AF. (Clinicaltrial.gov registration NCT00654290.). PMID- 19903190 TI - Regenerative therapy in peripheral artery disease. AB - Patients with peripheral artery disease (PAD) and critical limb ischemia are the main candidates for limb amputations and have a poor life expectancy. Frequently, these patients are not eligible for either surgical or percutaneous interventions aimed at mechanical revascularization. Therefore, new strategies need to be identified to offer these patients a viable therapeutic option. Gene and cell therapy hold great promise for the treatment of peripheral vascular diseases because, in animal models, local delivery of growth factors and endothelial progenitor cells result in new blood vessel formation and regeneration of ischemic tissues. In this article, are reviewed phase I and phase II gene, and cell therapy clinical trials in patients with PAD. PMID- 19903191 TI - Interferon-gamma and pulmonary macrophages contribute to the mechanisms underlying prolonged airway hyperresponsiveness. AB - BACKGROUND: Airway hyperresponsiveness (AHR) in asthmatics includes a variable component that persists following an allergen challenge. This may be dissociated from inflammatory cell recruitment, implying a role for resident pulmonary cells in regulating the response. OBJECTIVE: Using improved methods of assessing AHR in a mouse model of allergic airway disease, to investigate the basis of the development of prolonged AHR. METHOD: BALB/c mice were systemically sensitized and then challenged with aerosolized ovalbumin (OVA). Airway and tissue responsiveness were measured at baseline and at 1 day, and 1, 2 and 3 weeks after the last OVA challenge. Inflammatory cell numbers in BALF and levels of mRNA for eotaxin-1 and -2, IFN-gamma, IL-5 and -13 in the lung were measured at each time point. In further experiments, the roles of IFN-gamma and of CCR3(+) and CD4(+) cells in the development of prolonged AHR were assessed by blockade or depletion with monoclonal antibodies. The role of pulmonary macrophages was assessed by selective chemical depletion of these cells. RESULTS: Airway responsiveness was increased above baseline at 1 day after the last OVA challenge, and this was sustained for 1 week. In contrast, tissue-specific responsiveness was only significantly increased above baseline at 1 day. Development of prolonged AHR was inhibited by neutralization of IFN-gamma or by depletion of pulmonary macrophages, but not by depletion of either CD4(+) T cells or CCR3(+) eosinophils. CONCLUSION: An interaction between IFN-gamma and pulmonary macrophages contributed to the prolongation of airway hyperresponsiveness. In contrast, T cells and eosinophils did not contribute to prolongation of AHR. These findings emphasize the importance of the innate host response in the development of manifestations of asthma, as well as its potential relevance as a target for therapeutic intervention. PMID- 19903192 TI - Independent histological risk factors for lymph node metastasis of superficial esophageal squamous cell carcinoma; implication of claudin-5 immunohistochemistry for expanding the indications of endoscopic resection. AB - Endoscopic resection is curative for superficial esophageal squamous cell carcinoma (ESCC) limited to the lamina propria. Endoscopic resection is not recommended for superficial ESCC invading muscularis mucosa or submucosa, however, because of the high frequency of lymph node metastasis (LNM) in such patients. Methods to more accurately predict LNM by analysis of endoscopically resected specimens are needed. Patients with superficial ESCC who underwent surgery without prior chemoradiotherapy (n= 110) were retrospectively examined to determine whether LNM correlated with immunohistochemical parameters and conventional histological parameters, including depth of invasion and vascular permeation. Cancer cell expression of claudins-1, 5, and 7, E-cadherin, beta catenin, and matrix metalloproteinase 7 was evaluated. Univariate analysis revealed that LNM correlated with claudin-5 expression, but not any other immunohistochemical parameter examined. Multivariate analysis revealed three independent risk factors for LNM: aberrant claudin-5 expression in cancer cells (odds ratio; OR [95% confidence interval]= 4.61[1.44-14.77]), depth of submucosal invasion greater than 200 microm (3.55 [1.02-13.17]), and positive lymphatic permeation (3.34 [1.22-9.15]). LNM was found in one of 29 (3.4%) patients with none of these three risk factors, and in 32 of 81 (39.5%) patients with one or more of these risk factors. In superficial ESCC, routine analysis of claudin-5 expression in cancer cells together with depth of invasion and lymphatic permeation may be useful for predicting LNM and thereby reducing the number of patients undergoing additional surgery after successful endoscopic resection. PMID- 19903193 TI - Use of endoscopy in diagnosis and management of patients with dysphagia in an African setting. AB - The objectives of this study were to define the utility of esophagogastroduodenoscopy in the diagnosis and management of patients presenting with dysphagia and to determine the relative incidence of the various causes of dysphagia in Sudan. This is a prospective, cross-sectional, descriptive, hospital based study carried out at the endoscopy unit of Soba University Hospital, Khartoum, Sudan. All patients complaining of dysphagia underwent upper gastrointestinal endoscopy with therapeutic intervention when necessary. A total of 114 patients were enrolled in the study, with a mean age of 47 years SD +/- 19 and a male to female ratio of 1 : 1.04. A benign condition was diagnosed in 56% of the cases; this included esophageal strictures in 21% of the cases and achalasia in 14%. Malignant causes were mainly due to esophageal cancer (40.4%) and cancer of the stomach cardia (3.5%). Therapeutic intervention was attempted in 83% of the cases. Risk factors predictive of a malignant etiology were age over 40 years (P < 0.000), dysphagia lasting between 1 month and 1 year (P < 0.000), and weight loss (P < 0.000). A barium study was performed in 35 cases (31%) prior to endoscopic examination and proved to be inaccurate in three cases (8.6%). Upper gastrointestinal endoscopy in our African setting is an accurate and useful investigation in the diagnosis and management of patients presenting with dysphagia. Patients over the age of 40 years presenting with dysphagia and weight loss are more likely to have a neoplastic disease and should be referred for urgent endoscopy. PMID- 19903194 TI - Inversed Y cardioplasty plus a truncal vagotomy-antrectomy and a Roux-en-Y gastrojejunostomy performed in patients with stricture of the esophagogastric junction after a failed cardiomyotomy or endoscopic procedure in patients with achalasia of the esophagus. AB - Laparoscopic anterior cardiomyotomy in addition to anterior Dor's fundoplication is the procedure of choice for achalasia of the esophagus with approximately 95% success rate. Redo cardiomyotomy is complicated and associated with rerecurrence of dysphagia. Twelve patients with failed redo myotomy were clinically evaluated with radiology, endoscopy, and manometry in whom achalasia type III or IV was confirmed. We propose as treatment for these selected cases an inversed Y cardioplasty + truncal vagotomy, a partial distal gastrectomy and Roux-en-Y gastrojejunostomy in order to facilitate esophageal emptying and avoid the appearance of postoperative gastroesophageal reflux as a side effect of this procedure. One patient was reoperated on in order to enlarge the cardioplasty. Disappearance of dysphagia was confirmed in all patients. Three patients presented reflux symptoms and were treated with 20 mg of Omeprazole 20 twice/day. No food retention, erosive esophagitis, or Barrett's esophagus were observed. The mean resting pressure decreased from 24.9 +/- 8.5 mm Hg to 7.5 +/- 2.5 mm Hg (P = 0.0001). Furthermore, esophageal diameter decreased significantly after a 5-year follow-up. This procedure could be an option for treating patients in which repeated Heller operations have failed. PMID- 19903195 TI - The high incidence of esophageal cancer in parts of China may result primarily from genetic rather than environmental factors. AB - About 40,000 inhabitants migrated from a high-risk area of esophageal squamous cell carcinoma (ESCC) to a low-risk area of esophageal cancer 40 years ago. Little is known about the change in the mortality in esophageal cancer among these immigrants. This study examined the impact of changing environments on esophageal cancer by comparing age-standardized mortality rates of immigrant group to the rates of native population (natives who live in high cancer location and have never moved) and host populations (hosts who live in low cancer location and have never moved people). All ESCC deaths taking place during 1999-2004 among the migrant, native, and host populations were identified by retrospective population-based screening. Direct age-adjusted mortality rates were calculated by using the China population of year 2000 as standard population. From 1999 2004, the average annual age-adjusted mortality of ESCC for the migrant, native, and host population was 61.6/100,000, 59.7/100,000, and 6.7/100,000, respectively. No decreasing tendency was found in mortality rate of ESCC in the population of young immigrants. The mortality rate of ESCC of migrants remained high even they had been living in the low endemic region for 40 years. This study strongly suggested that genetic susceptibility, rather than environment exposure, is responsible for the high risk of ESCC in the migrants. PMID- 19903196 TI - Phylogenetic diversity metrics for ecological communities: integrating species richness, abundance and evolutionary history. AB - Phylogenetic information is increasingly being used to understand the assembly of biological communities and ecological processes. However, commonly used metrics of phylogenetic diversity (PD) do not incorporate information on the relative abundances of individuals within a community. In this study, we develop three indices of PD that explicitly consider species abundances. First, we present a metric of phylogenetic-abundance evenness that evaluates the relationship between the abundance and the distribution of terminal branch lengths. Second, we calculate an index of hierarchical imbalance of abundances at the clade level encapsulating the distribution of individuals across the nodes in the phylogeny. Third, we develop an index of abundance-weighted evolutionary distinctiveness and generate an entropic index of phylogenetic diversity that captures both information on evolutionary distances and phylogenetic tree topology, and also serves as a basis to evaluate species conservation value. These metrics offer measures of phylogenetic diversity incorporating different community attributes. We compare these new metrics to existing ones, and use them to explore diversity patterns in a typical California annual grassland plant community at the Jasper Ridge biological preserve. PMID- 19903198 TI - Highly expressed genes in a rough-colony-forming phenotype of Aggregatibacter actinomycetemcomitans: implication of a mip-like gene for the invasion of host tissue. AB - Aggregatibacter actinomycetemcomitans, a potent pathogen of periodontitis, typically grows as a rough and adherent colony on primary isolated cultures. The colony transforms into a smooth phenotype during repeated subculture. In this study, we aimed to identify highly expressed genes in the rough-colony-forming phenotype for isolation of host-induced genes. Using a cDNA-subtractive hybridization technique, three genes, homologous to a macrophage infectivity potentiator gene (mip), peroxiredoxin gene (prx) and outer membrane protein gene (ompA), were identified. The expression levels of these genes in the rough-colony forming phenotype were 4-10-fold higher as compared with the smooth-colony forming phenotype. Attention was focused on the mip-like gene, and a recombinant protein and a deficient mutant were constructed. The recombinant protein reacted with sera from patients with periodontitis, suggesting the production of the Mip like protein in periodontal lesions. Viable quantitative invasion assay demonstrated that the viable cell counts of the wild-type strain that invaded HeLa cells were more than fourfold as compared with the mip-deficient mutant. The expression of the mip-like gene, prx-like gene and ompA-like gene may be enhanced in the host, and the mip-like gene may play an important role in the infection of A. actinomycetemcomitans, especially in its invasion of the epithelium. PMID- 19903199 TI - Shifts in desulfonating bacterial communities along a soil chronosequence in the forefield of a receding glacier. AB - Forefields of receding glaciers are unique and sensitive environments representing natural soil chronosequences, where sulfate availability is assumed to be a limiting factor. Bacterial mineralization of organosulfur is an important sulfate-providing process in soils. We analyzed the diversity of sulfonate desulfurizing (desulfonating) bacteria in the Damma glacier forefield on the basis of the key gene asfA by terminal restriction fragment length polymorphism and clone libraries. The community structure and sequence diversity of desulfonating bacteria differed significantly between forefield soils deglaciated in the 1990s and the 1950s. Soil age had a strong effect on the desulfonating rhizosphere communities of Agrostis rupestris, but only a slight impact on the ones from Leucanthemopsis alpina. AsfA affiliated to Polaromonas sp. was predominantly found in the more recent ice-free soils and the corresponding rhizospheres of A. rupestris, while a group of unidentified sequences was found to be dominating the matured soils and the corresponding rhizospheres of A. rupestris. The desulfonating bacterial diversity was not affected by varying levels of sulfate concentrations. The level of asfA diversity in recently deglaciated soils suggests that desulfonating bacteria are a critical factor in sulfur cycling, with defined groups dominating at different stages of soil formation. PMID- 19903200 TI - The stress response protein Gls24 is induced by copper and interacts with the CopZ copper chaperone of Enterococcus hirae. AB - Intracellular copper routing in Enterococcus hirae is accomplished by the CopZ copper chaperone. Under copper stress, CopZ donates Cu(+) to the CopY repressor, thereby releasing its bound zinc and abolishing repressor-DNA interaction. This in turn induces the expression of the cop operon, which encodes CopY and CopZ, in addition to two copper ATPases, CopA and CopB. To gain further insight into the function of CopZ, the yeast two-hybrid system was used to screen for proteins interacting with the copper chaperone. This led to the identification of Gls24, a member of a family of stress response proteins. Gls24 is part of an operon containing eight genes. The operon was induced by a range of stress conditions, but most notably by copper. Gls24 was overexpressed and purified, and was shown by surface plasmon resonance analysis to also interact with CopZ in vitro. Circular dichroism measurements revealed that Gls24 is partially unstructured. The current findings establish a novel link between Gls24 and copper homeostasis. PMID- 19903201 TI - Expression of Escherichia coli Min system in Bacillus subtilis and its effect on cell division. AB - In both rod-shaped Bacillus subtilis and Escherichia coli cells, Min proteins are involved in the regulation of division septa formation. In E. coli, dynamic oscillation of MinCD inhibitory complex and MinE, a topological specificity protein, prevents improper polar septation. However, in B. subtilis no MinE is present and no oscillation of Min proteins can be observed. The function of MinE is substituted by that of an unrelated DivIVA protein, which targets MinCD to division sites and retains them at the cell poles. We inspected cell division when the E. coli Min system was introduced into B. subtilis cells. Expression of these heterologous Min proteins resulted in cell elongation. We demonstrate here that E. coli MinD can partially substitute for the function of its B. subtilis protein counterpart. Moreover, E. coli MinD was observed to have similar helical localization as B. subtilis MinD. PMID- 19903202 TI - Global analysis of mutual interaction surfaces of nucleosomes with comprehensive point mutants. AB - The surfaces of core histones in nucleosome are exposed as required for factor recognition, or buried for histone-DNA and histone-histone interactions. To understand the mechanisms by which nucleosome structure and function are coordinately altered in DNA-mediated reactions, it is essential to define the roles of both exposed and buried residues and their functional relationships. For this purpose, we developed GLASP (GLobal Analysis of Surfaces by Point mutation) and GLAMP (GLobal Analysis of Mutual interaction surfaces of multi-subunit protein complex by Point mutation) strategies, both of which are comprehensive analyses by point mutagenesis of exposed and buried residues in nucleosome, respectively. Four distinct DNA-mediated reactions evaluated by Ty suppression (the Spt(-) phenotype), and sensitivities to 6-azauracil (6AU), hydroxyurea (HU), and methyl methanesulfonate (MMS), require common and different GLAMP residues. Mutated GLAMP residues at the interface between histones H2A and H2B mainly affect the Spt(-) phenotype but not HU and MMS sensitivities. Interestingly, among the mutated GLAMP residues surrounding the histone H3-H3' interface, some equally affect the Spt(-) phenotype, and HU and MMS sensitivities, whereas others differentially affect the Spt(-) phenotype, and HU and MMS sensitivities. Based on these and other results, the functional relationships among chromatin factors and GLASP and GLAMP residues provide insights into nucleosome disassembly/assembly processes in DNA-mediated reactions. PMID- 19903205 TI - Preliminary population-based epidemiological and clinical data on 2009 pandemic H1N1 influenza A (pH1N1) from Lima, Peru. PMID- 19903206 TI - Concomitant administration of seasonal trivalent and pandemic monovalent H1N1 live attenuated influenza vaccines. PMID- 19903207 TI - Mortality burden of the 1918-1920 influenza pandemic in Hong Kong. PMID- 19903208 TI - Apropos 'performance of influenza point-of-care tests in the detection of pandemic (H1N1) 2009 influenza viruses'. PMID- 19903209 TI - Estimation of the reproductive number and the serial interval in early phase of the 2009 influenza A/H1N1 pandemic in the USA. AB - BACKGROUND: The United States was the second country to have a major outbreak of novel influenza A/H1N1 in what has become a new pandemic. Appropriate public health responses to this pandemic depend in part on early estimates of key epidemiological parameters of the virus in defined populations. METHODS: We use a likelihood-based method to estimate the basic reproductive number (R(0)) and serial interval using individual level U.S. data from the Centers for Disease Control and Prevention (CDC). We adjust for missing dates of illness and changes in case ascertainment. Using prior estimates for the serial interval we also estimate the reproductive number only. RESULTS: Using the raw CDC data, we estimate the reproductive number to be between 2.2 and 2.3 and the mean of the serial interval (mu) between 2.5 and 2.6 days. After adjustment for increased case ascertainment our estimates change to 1.7 to 1.8 for R(0) and 2.2 to 2.3 days for mu. In a sensitivity analysis making use of previous estimates of the mean of the serial interval, both for this epidemic (mu = 1.91 days) and for seasonal influenza (mu = 3.6 days), we estimate the reproductive number at 1.5 to 3.1. CONCLUSIONS: With adjustments for data imperfections we obtain useful estimates of key epidemiological parameters for the current influenza H1N1 outbreak in the United States. Estimates that adjust for suspected increases in reporting suggest that substantial reductions in the spread of this epidemic may be achievable with aggressive control measures, while sensitivity analyses suggest the possibility that even such measures would have limited effect in reducing total attack rates. PMID- 19903210 TI - Pediatric influenza-associated myositis - Nebraska, 2001-2007. AB - OBJECTIVE: Influenza-associated myositis (IAM), characterized by severe lower extremity myalgia and reluctance to walk, is a complication of influenza among children. We investigated IAM in Nebraska during six influenza seasons, 2001 2007. METHODS: During 2006-2007, we requested reports of severe influenza illness among persons aged <18 years and investigated medical records to identify and confirm IAM cases defined as severe myalgia with elevated serum creatinine kinase level in a patient aged <18 years, occurring within 7 days of laboratory confirmed influenza illness onset. Statewide hospital discharge data (HDD) were reviewed to identify retrospectively confirmed IAM cases during 2006-2007 and five previous seasons, by using surveillance data to define periods of influenza activity. Statewide IAM incidence was estimated for 2001-2002 through 2006-2007. RESULTS: During 2006-2007, a total of 13 IAM cases were confirmed by enhanced surveillance. Median age was 6 years (range, 4-11 years). Influenza diagnosis was established by viral isolation from six patients (one influenza A and five influenza B) and rapid diagnostic tests for seven. Twelve (92%) patients, including one who died, were hospitalized for a median of 3 days (range, 1-4 days). Review of HDD identified 12 retrospectively confirmed IAM cases during 2006-2007, including four not reported through enhanced surveillance, and only one during five previous seasons (2003-2004). The HDD-derived, retrospectively confirmed statewide IAM incidence estimates/100,000 population aged <18 years were 2.693 and 0.225 during 2006-2007 and 2003-2004, respectively. CONCLUSION: An IAM epidemic occurred in Nebraska during the 2006-2007 influenza season. PMID- 19903211 TI - Development of a new candidate H5N1 avian influenza virus for pre-pandemic vaccine production. AB - BACKGROUND: Highly pathogenic H5N1 avian influenza viruses currently circulating in birds have caused hundreds of human infections, and pose a significant pandemic threat. Vaccines are a major component of the public health preparedness for this likely event. The rapid evolution of H5N1 viruses has resulted in the emergence of multiple clades with distinct antigenic characteristics that require clade-specific vaccines. A variant H5N1 virus termed clade 2.3.4 emerged in 2005 and has caused multiple fatal infections. Vaccine candidates that match the antigenic properties of variant viruses are necessary because inactivated influenza vaccines elicit strain-specific protection. OBJECTIVE: To address the need for a suitable seed for manufacturing a clade 2.3.4 vaccine, we developed a new H5N1 pre-pandemic candidate vaccine by reverse genetics and evaluated its safety and replication in vitro and in vivo. METHODS: A reassortant virus termed, Anhui/PR8, was produced by reverse genetics in compliance with WHO pandemic vaccine development guidelines and contains six genes from A/Puerto Rico/8/34 as well as the neuraminidase and hemagglutinin (HA) genomic segments from the A/Anhui/01/2005 virus. The multi-basic cleavage site of HA was removed to reduce virulence. RESULTS: The reassortant Anhui/PR8 grows well in eggs and is avirulent to chicken and ferrets but retains the antigenicity of the parental A/Anhui/01/2005 virus. CONCLUSION: These results indicate that the Anhui/PR8 reassortant lost a major virulent determinant and it is suitable for its use in vaccine manufacturing and as a reference vaccine virus against the H5N1 clade 2.3.4 viruses circulating in eastern China, Vietnam, Thailand, and Laos. PMID- 19903212 TI - Genomic events underlying the changes in adamantane resistance among influenza A(H3N2) viruses during 2006-2008. AB - BACKGROUND: Adamantanes resistance in H3N2 viruses has been increasing since 2000, and in 2005-2006 reached nearly 100% in most countries, with the circulation of the N-lineage. In 2006-2007, however, a significant decrease in resistance was observed in many regions. OBJECTIVES: To explore potential links between adamantane resistance and the A(H3N2) viruses that circulated between 2006 and 2008. METHODS: A total of 1451 Influenza A (H3N2) viruses collected globally in 2001-2008 were screened for the presence of adamantane resistance markers. A subset of 100 viruses representing the broad genetic and geographic spectrum of these viruses was selected for complete genome sequencing and phylogenetic analyses. RESULTS: Full genome sequence analysis of 2006-2007 viruses revealed co-circulation of four distinct genotypes, designated A-D. Phylogenetic analyses demonstrated reassortment between viruses from the N lineage and other viruses that had circulated in prior seasons, including those bearing an adamantane sensitive marker. Genotype D viruses became dominant in late 2006-2007 and continued to be the main H3N2 genotype in 2007-2008. Viruses of this genotype retained all N-lineage genome segments except PB2 and NP, which were acquired through reassortment. CONCLUSIONS: The decrease in adamantane resistance at that time was due to transient co-circulation of genotypes that emerged through reassortment. Our findings emphasize the importance of complete genome sequencing in understanding the complex nature of the relationship between influenza virus evolution and antiviral resistance. The recent emergence of the pandemic multi-reassortant H1N1 virus underscores the importance of whole genome sequence monitoring for rapid detection of such unusual and novel strains. PMID- 19903213 TI - Safety and immunogenicity of an inactivated thimerosal-free influenza vaccine in infants and children. AB - OBJECTIVE: Few prospective studies of inactivated split virion influenza vaccine have been conducted in infants and children. Our objective was to evaluate the safety, reactogenicity and immunogenicity of a thimerosal-free inactivated influenza vaccine (Fluvax; CSL Limited, Parkville, Australia) in children aged 6 months to <9 years. METHODS: A prospective, open-label, phase III clinical trial was conducted in 298 healthy children previously unvaccinated with influenza, commencing in the Southern Hemisphere 2005 autumn. Participants were divided into two groups (Group A: > or =6 months to <3 years; Group B: > or =3 years to <9 years), and received two doses of the 2005 vaccine, and one dose of the 2006 vaccine one year later (Group A: 0.25 ml per dose; Group B: 0.5 ml per dose). Vaccine safety and reactogenicity was evaluated for 30 days after each dose. Immunogenicity was assessed using hemagglutination inhibition and single radial hemolysis assays. RESULTS: There were no withdrawals due to adverse events (AEs). The majority of solicited local and systemic AEs were of mild severity. A maximum intensity of severe was reported for injection site pain and fever by only 3.0% and 3.4% of participants, respectively. The vaccine was immunogenic for all antigens, with > or =95% of both younger and older children achieving seroprotection after dose 2. CONCLUSIONS: This thimerosal-free inactivated influenza vaccine had a favorable safety profile and was immunogenic in children aged > or =6 months and <9 years. Primary and booster vaccination produced consistently immunogenic responses including in children under 3 years of age receiving 0.25 ml doses of vaccine. PMID- 19903214 TI - Molecular characterization of adenovirus circulating in Central and South America during the 2006-2008 period. AB - BACKGROUND: Human Adenoviruses are recognized pathogens, causing a broad spectrum of diseases. Serotype identification is critical for epidemiological surveillance, detection of new strains and understanding of HAdvs pathogenesis. Little data is available about HAdvs subtypes in Latin America. METHODS: In this study, we have molecularly characterized 213 adenoviruses collected from ILI presenting patients, during 2006-08, in Central and South America. RESULTS: Our results indicate that 161(76%) adenoviruses belong to subgroup C, 45 (21%) to subgroup B and 7 (3%) to subtype E4. PMID- 19903215 TI - Palisaded neutrophilic granulomatous dermatitis with leukocytoclastic vasculitis in a patient without any underlying systemic disease detected to date. AB - Palisaded neutrophilic granulomatous dermatitis (PNGD) is a rare dermatologic condition which shows various clinical and histopathological features. Although the PNGD lesions have been suggested to begin as leukocytoclastic vasculitis (LCV), there is still insufficient clinicopathological information in the reported cases of PNGD in acute stage with LCV. The relationship between PNGD and interstitial granumatous dermatis (IGD) also remains unclear. This report presents the case of a 60-year-old female patient with multiple erythematous nodules on the extremities. She had no underlying systemic disease detected to date, although transient, abnormal liver function tests were seen. The histopathological examination of an erythematous nodule revealed the features of PNGD in the acute stage. The patient presented the characteristic features of LCV including palisaded granulomatous pattern, and the interstitial granulomatous pattern was seen together, suggesting that PNGD with LCV can show an interstitial granulomatous pattern. The present case also suggested that PNGD in the acute stage with LCV tends to clinically manifest as erythematous nodules on the extremities and histopathologically shows a remarkable papillary edema and an extensive fibrin deposition in and around the vessel wall. PNGD may be associated with transient liver dysfunction. PMID- 19903216 TI - Necrotizing palisaded granulomatous dermatitis as a manifestation of familial hemophagocytic lymphohistiocytosis. PMID- 19903217 TI - Extragenital lichen sclerosus et atrophicus mimicking cutaneous T-cell lymphoma: report of a case. AB - Early lesions of lichen sclerosus et atrophicus (LSA) may present as a mild lichenoid tissue reaction, occasionally together with basilar epidermotropism, mimicking early cutaneous T-cell lymphoma, mycosis fungoides (MF) variant. We report a case of extragenital LSA in which both histological patterns were present in the same clinically homogenous and stable lesion. A 27-year-old man presented with a history of white atrophic plaques on the trunk. A biopsy of an abdominal lesion revealed epidermal thinning, a superficial perivascular lymphoid cell infiltrate with focal epidermotropism, mild nuclear atypia and perinuclear halos. Immunophenotyping showed decreased CD5 and CD7, with a slight predominance of CD8-positive T-lymphocytes. All these changes were suggestive of MF. However, a repeat biopsy 3 months later from the same stable plaque revealed features diagnostic of LSA. LSA mimicking early MF histologically has been reported in genital skin. Conversely, MF may clinically and histopathologically resemble LSA. With gene rearrangement studies, clonal proliferation may not be detected in early MF but has been reported to occur in LSA. Awareness of the histopathologic spectrum of LSA within a stable plaque is important to avoid a potential diagnostic pitfall, and should prompt a repeat biopsy. PMID- 19903218 TI - Cutaneous presentation of post-renal transplant lymphoproliferative disorder: a series of four cases. AB - We report detailed histological and molecular characteristics of four post transplant lymphoproliferative disorders (PTLD) presenting in the skin of renal transplant patients, and their clinical outcome. Three had B-cell lymphomas (cases 1-3), and one had a T-cell lymphoma (case 4). All B-cell lymphomas showed Epstein-Barr virus (EBV) by immunohistochemistry (IHC) or in situ hybridization (ISH). Cases 1 and 2 were large cell lymphomas, and case 3 a plasmacytoma. Case 1 showed light chain restriction and heavy chain gene rearrangement by polymerase chain reaction (PCR). The patient was then diagnosed with an abdominal lymphoma and died of sepsis. Case 2 had no recoverable DNA. Case 3 had a plasmacytoma that showed monoclonal light chain restriction on IHC and an oligoclonal heavy chain rearrangement by PCR. In cases 2 and 3, the lesions regressed following reduction of immunosuppression, and died 1.5 and 8 years later from unrelated medical causes. Case 4 was a CD 30+ anaplastic large T-cell lymphoma with no EBV detected by IHC, ISH and PCR, and died of heart failure 2 years later. Cutaneous manifestations of PTLD are rare, show wide array of clinical and pathological features, and generally have a favorable prognosis. EBV appears to be associated only with B-cell cutaneous lymphomas. PMID- 19903219 TI - The evolution of haplodiploidy by male-killing endosymbionts: importance of population structure and endosymbiont mutualisms. AB - Haplodiploid inheritance systems, characterized by male transmission of only their maternally inherited genomic elements, have evolved more than 20 times within the animal kingdom. A number of theoretical studies have argued that infection with certain male-killing endosymbionts can potentially lead to the evolution of haplodiploidy. By explicitly investigating the coevolutionary dynamics between host and endosymbiont, we show that the assumptions of current models cannot explain the evolution of haplodiploidy very well, as the endosymbiont will often go extinct in the long term. Here, we provide two additional mechanisms that can explain the stable evolution of haplodiploidy by male-killing endosymbionts. First of all, a spatially structured population can facilitate the long-term persistence of haplodiploidy, but this applies only when levels of inbreeding are very high. By contrast, endosymbionts that are mutualistic with their hosts provide a much more general and promising route to the stable evolution of haplodiploidy. This model is the first to provide a formal explanation of the supposed association between the evolution of haplodiploidy and the highly inbred lifestyles of some ancestors, while it also provides a hypothesis for the evolution of haplodiploidy in more outbred ancestors. PMID- 19903220 TI - Construction and application of efficient Ac-Ds transposon tagging vectors in rice. AB - Transposons are effective mutagens alternative to T-DNA for the generation of insertional mutants in many plant species including those whose transformation is inefficient. The current strategies of transposon tagging are usually slow and labor-intensive and yield low frequency of tagged lines. We have constructed a series of transposon tagging vectors based on three approaches: (i) AcTPase controlled by glucocorticoid binding domain/VP16 acidic activation domain/Gal4 DNA-binding domain (GVG) chemical-inducible expression system; (ii) deletion of AcTPase via Cre-lox site-specific recombination that was initially triggered by Ds excision; and (iii) suppression of early transposition events in transformed rice callus through a dual-functional hygromycin resistance gene in a novel Ds element (HPT-Ds). We tested these vectors in transgenic rice and characterized the transposition events. Our results showed that these vectors are useful resources for functional genomics of rice and other crop plants. The vectors are freely available for the community. PMID- 19903221 TI - Molecular characterization of a dehydroascorbate reductase from Pinus bungeana. AB - Abstract Dehydroascorbate reductase (DHAR) plays a critical role in the ascorbate glutathione recycling reaction for most higher plants. To date, studies on DHAR in higher plants have focused largely on Arabidopsis and agricultural plants, and there is virtually no information on the molecular characteristics of DHAR in gymnosperms. The present study reports the cloning and characteristics of a DHAR (PbDHAR) from a pine, Pinus bungeana Zucc. ex Endl. The PbDHAR gene encodes a protein of 215 amino acid residues with a calculated molecular mass of 24.26 kDa. The predicted 3-D structure of PbDHAR showed a typical glutathione S-transferase fold. Reverse transcription-polymerase chain reaction revealed that the PbDHAR was a constitutive expression gene in P. bungeana. The expression level of PbDHAR mRNA in P. bungeana seedlings did not show significant change under high temperature stress. The recombinant PbDHAR was overexpressed in Escherichia coli following purification with affinity chromatography. The recombinant PbDHAR exhibited enzymatic activity (19.84 micromol/min per mg) and high affinity (a K(m) of 0.08 mM) towards the substrates dehydroascorbate (DHA). Moreover, the recombinant PbDHAR was a thermostable enzyme, and retained 77% of its initial activity at 55 degrees C. The present study is the first to provide a detailed molecular characterization of the DHAR in P. bungeana. PMID- 19903222 TI - Dehydration kinetics of embryonic axes from desiccation-sensitive seeds: an assessment of descriptive models. AB - The response of desiccation-sensitive plant tissues to dehydration is significantly affected by dehydration conditions, particularly the rate of drying. Consequently it is important to be able to quantify drying rate. The aim of the study was to assess two models that have been proposed to describe drying kinetics, and thus to provide a quantification of non-linear drying rates, of embryonic axes excised from recalcitrant seeds. These models are an exponential drying time course, and a modified inverse relationship, respectively. For the six species investigated here the inverse function was generally found to fit drying data better than the exponential function under both rapid and slow drying conditions, and so is recommended. The rate of drying, under the conditions used here, was determined by axis size and possibly the nature of the axis outer coverings, rather than the water activity difference between the tissue and surrounding air. PMID- 19903223 TI - Mobilization and acquisition of sparingly soluble P-sources by Brassica cultivars under P-starved environment I. Differential growth response, P-efficiency characteristics and P-remobilization. AB - Phosphorus (P) starvation is highly notorious for limiting plant growth around the globe. To combat P-starvation, plants constantly sense the changes in their environment, and elicit an elegant myriad of plastic responses and rescue strategies to enhance P-solublization and acquisition from bound soil P-forms. Relative growth responses, P-solublization and P-acquisition ability of 14 diverse Brassica cultivars grown with sparingly soluble P-sources (Rock-P (RP) and Ca(3)(PO(4))(2) (TCP)) were evaluated in a solution culture experiment. Cultivars showed considerable genetic diversity in terms of biomass accumulation, concentration and contents of P and Ca in shoots and roots, P-stress factor (PSF) and P use efficiency. Cultivars showed variable P-stress tolerance, and cultivars depicting low PSF and high P-efficiency values were better adaptable to P starvation. In experiment 2, after initial feeding on optimum nutrition for 12 d after transplanting (DAT), class-I (low P-tolerant (Oscar and Con-II)) and class II (low P-sensitive (Gold Rush and RL-18)) cultivars were exposed to P-free environment for 25 d. All of the cultivars remobilized P from above ground parts to their roots during growth in P-free environment, the magnitude of which was variable in tested cultivars. P-concentrations ([P]s) at 37 DAT were higher in developing compared with developed leaves. Translocation of absorbed P from metabolically inactive to active sites in P-stressed plants may have helped class I cultivars to establish a better rooting system, which provided a basis for enhanced P-utilization efficiency (PUE) and tolerance against P-stress. By supplying TCP and RP spatially separated from other nutrients in split root study, class-I cultivars were still able to mobilize RP and TCP more efficiently compared with class-II cultivars. To compare the growth behavior under P-stress, cultivars were grown in pots for 41 d after sowing, using a soil low in P (NaHCO(3)-extractable P = 3.97 mg/kg, Mehlich-III-extractable P = 6.13 mg/kg) with (+P = 60 mg P/kg soil) or without P addition (0P) in study 4. Tested cultivars showed genetic diversity in PUE, P-efficiency (PE), P-efficiency ratio (PER) and PSF. P-stress markedly reduced biomass and plant P contents. Cultivars that produced higher root biomass accumulated higher total P-contents (r= 0.98**), which in turn was related negatively to PSF (r=-0.95**) and positively to shoot and total biomass. PER and PE showed significant correlations with shoot P-contents and biomass. Cultivars depicting high PUE and PE, and low PSF values showed better growth behavior under low soil P-environment. Systematic analysis and deployment of the plant rescue traits underlying the nutrient acquisition, assimilation, utilization and remobilization under P-starvation will bring more sparingly soluble P into cropping systems and will help to scavenge more P from plant unavailable bound P reserves. PMID- 19903225 TI - Substrate selectivity of glycerol-3-phosphate acyl transferase in rice. AB - Substrate selectivity of glycerol-3-phosphate acyltransferase (EC 2. 3. 1. 15) of rice (Oryza sativa L.) was explored in a comparative study of acyltransferases from seven plant species. In vitro labeling of acyl carrier protein (ACP) with (14)C or (3)H showed that acyltransferase from chill-sensitive plants, such as rice that uses either oleic (18:1) or palmitic acid (16:0) as acyl donor at comparable rates, displays lower selectivity than the enzyme from chill-resistant plants, such as spinach, which preferentially uses oleic acid (18:1) rather than palmitic acid (16:0) as an acyl donor. This may be a result of the size and character of the substrate-binding pocket of acyltransferase. Homology modeling and protein structure-based sequence alignment of acyltransferases revealed that proteins from either chill-sensitive or chill-tolerant plants shared a highly conserved domain containing the proposed substrate-binding pocket. However, the aligned residues surrounding the substrate-binding pocket are highly heterogeneous and may have an influence mainly on the size of the substrate binding pockets of acyltransferases. The substrate selectivity of acyltransferase of rice can be improved by enlarging the substrate-binding pocket using molecular biological methods. PMID- 19903224 TI - Mobilization and acquisition of sparingly soluble P-Sources by Brassica cultivars under P-starved environment II. Rhizospheric pH changes, redesigned root architecture and pi-uptake kinetics. AB - Non-mycorrhizal Brassica does not produce specialized root structures such as cluster or dauciform roots but is an effective user of P compared with other crops. In addition to P-uptake, utilization and remobilization activity, acquisition of orthophosphate (Pi) from extracellular sparingly P-sources or unavailable bound P-forms can be enhanced by biochemical rescue mechanisms such copious H(+)-efflux and/or carboxylates exudation into rhizosphere by roots via plasmalemma H(+) ATPase and anion channels triggered by P-starvation. To visualize the dissolution of sparingly soluble Ca-phosphate (Ca-P), newly formed Ca-P was suspended in agar containing other essential nutrients. With NH(4)(+) applied as the N source, the precipitate dissolved in the root vicinity can be ascribed to rhizosphere acidification, whereas no dissolution occurred with nitrate nutrition. To observe in situ rhizospheric pH changes, images were recorded after embedding the roots in agar containing bromocresol purple as a pH indicator. P-tolerant cultivar showed a greater decrease in pH than the sensitive cultivar in the culture media (the appearance of typical patterns of various colors of pH indicator in the root vicinity), and at stress P-level this acidification was more prominent. In experiment 2, low P-tolerant class-I cultivars (Oscar and Con-II) showed a greater decrease in solution media pH than low P-sensitive class-II (Gold Rush and RL-18) cultivars, and P-contents of the cultivars was inversely related to decrease in culture media pH. To elucidate P stress-induced remodeling and redesigning in a root architectural system, cultivars were grown in rhizoboxes in experiment 3. The elongation rates of primary roots increased as P-supply increased, but the elongation rates of the branched zones of primary roots decreased. The length of the lateral roots and topological index values increased when cultivars were exposed to a P-stress environment. To elucidate Pi-uptake kinetics, parameters related to P influx: maximal transport rate (V(max)), the Michaelis-Menten constant (K(m)), and the external concentration when net uptake is zero (C(min)) were tested in experiment 4. Lower K(m) and C(min) values were better indicative of the P-uptake ability of the class-I cultivars, evidencing their adaptability to P-starved environmental cues. In experiment 5, class-I cultivars exuded two- to threefold more carboxylates than class-II cultivars under the P-stress environment. The amount and types of carboxylates exuded from the roots of P-starved plants differed from those of plants grown under P-sufficient conditions. Nevertheless, the exudation rate of both class-I and class-II cultivars decreased with time, and the highest exudation rate was found after the first 4 h of carboxylates collection. Higher P uptake by class-I cultivars was significantly related to the drop in root medium pH, which can be ascribed to H(+)-efflux from the roots supplied with sparingly soluble rock-P and Ca(3)(PO(4))(2). These classical rescue strategies provided the basis of P-solubilization and acquisition from sparingly soluble P-sources by Brassica cultivars to thrive in a typically stressful environment. PMID- 19903229 TI - Introduction: Themed issue - papers arising from 7th Polish-Japanese Joint Seminar on Micro and Nano Analysis. PMID- 19903226 TI - Water supply changes N and P conservation in a perennial grass Leymus chinensis. AB - Changes in precipitation can influence soil water and nutrient availability, and thus affect plant nutrient conservation strategies. Better understanding of how nutrient conservation changes with variations in water availability is crucial for predicting the potential influence of global climate change on plant nutrient use strategy. Here, green-leaf nitrogen (N) and phosphorus (P) concentrations, N- and P-resorption proficiency (the terminal N and P concentration in senescent leaves, NRP and PRP, respectively), and N- and P-resorption efficiency (the proportional N and P withdrawn from senescent leaves prior to abscission, NRE and PRE, respectively) of Leymus chinensis (Trin.) Tzvel., a typical perennial grass species in northern China, were examined along a water supply gradient to explore how plant nutrient conservation responds to water change. Increasing water supply at low levels (< 9000 mL/year) increased NRP, PRP and PRE, but decreased green leaf N concentration. It did not significantly affect green-leaf P concentration or NRE. By contrast, all N and P conservation indicators were not significantly influenced at high water supply levels (> 9000 mL/year). These results indicated that changes in water availability at low levels could affect leaf-level nutrient characteristics, especially for the species in semiarid ecosystems. Therefore, global changes in precipitation may pose effects on plant nutrient economy, and thus on nutrient cycling in the plant-soil systems. PMID- 19903230 TI - Fine structures of wing scales in Sasakia charonda butterflies as photonic crystals. AB - We investigate the microstructure of scales in the wings of male Sasakia charonda charonda butterflies by scanning electron microscopy with the aid of optical microscopy. Six types of scales are identified: B1, W1 and R1 in brown background yellow spots and red spots, respectively; B2 in iridescent purple-blue and W2 in white pearl, both of which characterize the male and B3 in the wing edges. The B1, W1 and R1 scales are almost the same in structure and the B2 and W2 scales are almost the same. The difference among the B, W and R scales is in species and content of pigment. The B1, W1 and R1 scales have only two layers of cuticle lapped on the ridges. In contrast with them, the B2 and W2 scales have seven multilayers of cuticle piled on the ridge. The multiple interference of light that occurs among these cuticle layers, spaced with air layers, generates the significant iridescence of the B2 and W2 scales. Thus, the characteristic purple blue of the male wings is ascribed to the combination of the structural and chemical colouration in the B2 scales with melanin. The photonic crystals of these scales may be applicable to fine light manipulators such as reflection elements in laser diodes. B3 has many holes between the ridges and no multilayers of cuticle on the ridges. These structures may play any role in aerodynamically easy flight and/or in drainage of wet wings. PMID- 19903231 TI - Electron microscopy study of L1(0)-FePtCu nanoparticles synthesized at 613K. AB - Finely dispersed hard magnetic L1o-FePtCu nanoparticles with 100 orientation were directly synthesized by RF-sputtering on NaCl substrate at a temperature of 613 K. The maximum coercivity of the particles was 1.4 kOe (at RT). Degrees of atomic long-range order (LRO) for the L1o-FePtCu nanoparticles with different sizes were obtained using nanobeam electron diffraction technique. The decrease of LRO parameter became remarkable when the size became below 8 nm. The coercivity value also decreased with decreasing the particle size. The relation between the LRO parameter decrease and the coercivity decrease with particle size was discussed. PMID- 19903232 TI - HAADF-STEM analysis of layered double perovskite La(2)CuSnO(6) grown epitaxially. AB - Structural observation of layered double perovskite oxide La(2)CuSnO(6) thin films grown epitaxially on SrTiO(3) is reported by high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM). Particularly the transition layer at the interface was observed, and the first B site layer at the interface was found to be almost formed by the Cu atomic layer as the random structure, followed by formation of the layered structure. In addition, HAADF STEM images indicate that the thin film is not single crystalline, but some irregular structures were observed to grow around the interface near atomic steps of the substrate of SrTiO(3). Therefore, the steps largely affect the growth process of the thin film. PMID- 19903233 TI - Microstructure analysis of Nd-Fe-B sintered magnets improved by Tb-metal vapour sorption. AB - Behaviours of constituent elements in the Nd-Fe-B sintered magnets improved by Tb metal vapour sorption have been investigated by using an analytical transmission microscopy. It was found that a triple junction of the grain boundaries consists of fine Nd-O crystalline and amorphous phase. The energy dispersive X-ray spectroscopy (EDS) analysis showed that the amorphous phase mainly consists of Co, Nd and Tb. The Tb-treatment causes the formation of the amorphous Co-Nd(Tb) wetting-layer phase which wraps each Nd(2)Fe(14)B grain. The results suggest that the wrapped structure prevents the nucleation of magnetic reversed domains and then improves significantly the coercivity of the magnet. PMID- 19903234 TI - Neutron scattering studies of BiFeO(3) multiferroics: a review for microscopists. AB - Application of the neutron scattering technique in the study of crystal and magnetic properties of multiferroic BiFeO(3) is presented. The crucial role of the neutron scattering technique, complementary to X-ray diffraction method and transmission electron microscopy, is shown. Especially the ultra high-resolution time-of-flight (TOF) neutron diffraction technique used by Sosnowska et al. to detect the magnetic cycloid ordering and its role in studies of physical properties of BiFeO(3) and its alloys are reviewed. The first inelastic neutron scattering patterns of magnetic excitations in BiFeO(3) are also presented. Applications of different microscopy techniques such as transmission electron microscopy (TEM), scanning electron microscopy (SEM), field emission TEM and SEM (FESEM and FETEM), magnetic force microscope (MFM) and polarization force microscopy (PFM) bring insight on the fundamental problem of ferroelectricity and confirm the potential of BiFeO(3) multiferroic material for nanoscale devices. PMID- 19903235 TI - TEM determination of directions of (Ga,Mn)As nanowires grown by MBE on GaAs(001) substrates. AB - The structure of GaMnAs nanowires (NW) with nominal Mn concentration of up to 7 at% was investigated by transmission electron microscopy. The (Ga,Mn)As NW were grown on epiready GaAs(001) n-type wafers by molecular beam epitaxy. The crystal structure of the NW was determined to be zinc-blende. NW with Mn concentrations lower than 5 at% grow along the 111 direction. NW with higher Mn concentrations grow along the 110 direction and reveal a branching structure. The main nanowire and branches grow along the 110 directions belonging to only one {111} plane. PMID- 19903236 TI - Microstructure and properties of hot compacted powders of aluminium alloys. AB - Atomized 6061 aluminium alloy powders with and without the addition of 2 wt% Zr were milled for 80 h in a planetary ball mill and hot pressed in vacuum. The milled powders showed microhardness of about 170 HV, which increased after hot pressing up to 260 HV and up to 280 HV for powders without and with the Zr additions, respectively. Compression tests showed the high yield stress of 300 MPa obtained for the hot-pressed sample produced from the initial powders compared with ultimate compression strength of above 800 MPa for that of the milled sample and slightly higher for that with Zr additions. The effect of hot pressing on the structure of powders was investigated using a conventional analytical and high-resolution electron microscopy and high angle annular dark field scanning transmission electron microscopy combined with energy dispersive X ray microanalysis. The samples of initial powders hot pressed in vacuum showed a cell structure with particles of the Mg(2)Si and AlFeSi phases in intercell areas. In the milled and hot-pressed sample, the homogeneous structure of small grains of size below 200 nm was observed. The AlFeSi and Mg(2)Si particles with size 20-100 nm were uniformly distributed as well as the Zr rich particles in the Zr containing alloy. The Zr-rich particles containing up to 80 at% Zr were identified as a metastable fcc cubic phase with lattice parameter a= 0.48 nm. PMID- 19903237 TI - Formation process of beta-FeSi from amorphous Fe-Si synthesized by ion implantation: Fe concentration dependence. AB - Formation processes of beta-FeSi(2) from amorphous Fe-Si layers have been investigated using transmission electron microscopy (TEM). Si(111) substrates were irradiated with 120 keV Fe ions at -150 degrees C to fluences of 1.0 x 10(17) and 4.0 x 10(17) cm(-2). An amorphous Fe-Si layer embedded in an amorphous Si was formed in the low-fluence sample, whereas an amorphous Fe-Si surface layer on an amorphous Si was obtained in the high-fluence one. The amorphous Fe-Si layers were crystallized to beta-FeSi(2) after thermal annealing at 800 degrees C for 2 h. Cross-sectional and plan-view TEM observations revealed that, prior to the formation of beta-FeSi(2), the amorphous Fe-Si layers crystallized to alpha FeSi(2) in the low-fluence sample and to epsilon-FeSi in the high-fluence one. The absence of metastable gamma-FeSi(2) which is considered as a precursor of epitaxially grown beta-FeSi(2) on Si was attributed to the instability of gamma phase in an amorphous matrix. PMID- 19903238 TI - Local analysis of the edge dislocation core in BaTiO(3) thin film by STEM-EELS. AB - The a <100> edge dislocation core formed in an epitaxial BaTiO(3) (BTO) thin film grown on a substrate was investigated by scanning transmission electron microscopy combined with electron energy-loss spectroscopy. Elemental analysis using core-loss spectrum indicates that the atomic ratios of O/Ti and Ba/Ti are decreased at the dislocation core. The near-edge fine structure of the oxygen K edge recorded from the dislocation core differs slightly from that of relaxed BTO region, which suggests that Ba-O bonding is decreased at the dislocation core. The structure of the dislocation core is discussed using a high-angle annular dark-field image and the electron energy-loss spectroscopy results. PMID- 19903239 TI - HRTEM studies of NiNbZr + Ag amorphous-nanocrystalline composites. AB - Amorphous powder of composition corresponding to Ni60Ti20Zr20 (in at%) was obtained by ball milling in a high-energy mills starting from pure elements. Formation of the amorphous structure was observed already after 20 h of milling, although complete amorphization occurred after 40 h. The microhardness of powders increased from about 30 HV for pure elements to above 400 HV (1290 MPa) after 40 h of milling. Transmission electron microscopy (TEM) allowed to identify nanocrystalline inclusions of intermetallic phases of size 2-10 nm. Uniaxial hot pressing was performed in vacuum at temperature below the crystallization T(x) it is 510 degrees C and pressure of 600 MPa, Mixed amorphous powders and nanocrystalline silver powders were used to form a composite, in which microhardness was near 970 MPa HV and 400 HV for the amorphous phase and nanocrystalline silver, respectively. The compression strength of the composite containing 20 wt% of nanocrystalline Ag powder was equal to 600 MPa and plastic strain was 2%. Microstructure studies showed low porosity of composites of less than 1%, uniform distribution of the silver phase and a transition zone between both components, about 150 nm thick, where diffusion of nickel, niobium and zirconium into silver was observed. High-resolution TEM allowed identifying the structure of nanocrystalline inclusions in the amorphous matrix after hot pressing as either Ni(3)Zr or Ni(17)Nb(3). The identification was performed basing on measurements of angles and interatomic distances using inverse Fourier transformed images with enhanced contrast using Digital Micrograph computer program. PMID- 19903240 TI - New approach to cathodoluminescence studies in application to InGaN/GaN laser diode degradation. AB - Cathodoluminescence (CL) studies are widely applied in semi-conductor science and technology. However, for structures with a p-n junction the CL spatial distribution can be strongly affected by internal current flows of the electron beam induced current generated within the structure. This influence is the investigated in application to CL studies of degradation in aged laser diodes with InGaN multiquantum wells. PMID- 19903241 TI - Microstructure of ball milled and compacted Co-Ni-Al alloys from the beta range. AB - Two powder alloys from the beta phase region of compositions Co(28.5)Ni(36.5)Al(35) and Co(35)Ni(30)Al(35) were ball milled for 80 h in a high energy ball mill. The formation of amorphous structure was observed after 40 h of milling and further milling did not change their structure. The analytical and high-resolution transmission electron microscopy (TEM, HREM) examination of powder structure showed that nanoparticles of L1(0) phase of size of about 5 nm were present within the amorphous matrix. The vacuum hot pressing of the milled powders under pressure of 400 MPa at 700 degrees C for 12 min resulted in the formation of compacts with density of about 70% of the theoretical one. The additional heat treatment at 1300 degrees C for 6 h followed by water quenching, led to significant improvement of density and induced the martensitic transformation manifested by a broad heat effect. The characteristic temperatures of the transformation were determined using DSC measurements, which revealed only small differences within the examined alloys compositions. TEM structure studies of heat-treated alloys allowed to identify the structure of an ordered beta (B2) phase and L1(0) martrensite. PMID- 19903242 TI - TEM, HRTEM, electron holography and electron tomography studies of gamma' and gamma'' nanoparticles in Inconel 718 superalloy. AB - The aim of the study was the identification of gamma' and gamma'' strengthening precipitates in a commercial nickel-base superalloy Inconel 718 (Ni-19Fe-18Cr-5Nb 3Mo-1Ti-0.5Al-0.04C, wt %) using TEM dark-field, HRTEM, electron holography and electron tomography imaging. To identify gamma' and gamma'' nanoparticles unambiguously, a systematic analysis of experimental and theoretical diffraction patterns were performed. Using HRTEM method it was possible to analyse small areas of precipitates appearance. Electron holography and electron tomography techniques show new possibilities of visualization of gamma' and gamma'' nanoparticles. The analysis by means of different complementary TEM methods showed that gamma'' particles exhibit a shape of thin plates, while gamma' phase precipitates are almost spherical. PMID- 19903243 TI - Up-regulation of the type 3 ryanodine receptor is neuroprotective in the TgCRND8 mouse model of Alzheimer's disease. AB - The cellular pathology of Alzheimer's disease is progressive and protracted leading eventually to considerable neuronal death. The underlying mechanisms of the pathology are complex but changes in the control of intracellular Ca2+ are believed to contribute to the demise of neurons. In this study, we investigated the functional consequences of an increase in the expression of the type 3 isoform of the ryanodine receptor (RyR3). We found that although cortical neurons from TgCRND8 mice secreted significantly more amyloid beta protein and showed significantly increased RyR3 expression, they were no more sensitive to cell stress than non-transgenic neurons. Furthermore, despite increased intracellular Ca2+ release in response to ryanodine, we found that basal Ca2+, K+-evoked Ca2+ responses, and capacitative Ca2+ entry were no different in TgCRND8 neurons compared with non-transgenic neurons. Therefore, as RyR3 up-regulation did not affect neuronal health or global Ca2+ homeostasis, we investigated the effect of reducing RyR3 expression using small interfering RNA. Surprisingly, a reduction of RyR3 expression in TgCRND8, but not in non-transgenic, neurons increased neuronal death. These data reveal a new role for RyR3 and indicate a novel potential therapeutic target to delay or prevent the progression of Alzheimer's disease. PMID- 19903244 TI - ATRA inhibits ceramide kinase transcription in a human neuroblastoma cell line, SH-SY5Y cells: the role of COUP-TFI. AB - Ceramide is the central lipid in the sphingolipid metabolism. Ceramide kinase (CERK) and its product, ceramide 1-phosphate, have been implicated in various cellular functions. However, the regulatory mechanism of CERK gene expression remains to be determined. Here, we examined CERK mRNA level during all-trans retinoic acid (ATRA)-induced differentiation of a human neuroblastoma cell line, SH-SY5Y. ATRA reduced CERK mRNA and protein levels. Over-expression and small interfering RNA (siRNA) of CERK revealed that CERK is inhibitory against ATRA induced neuronal differentiation and cell growth arrest. ATRA inhibited the transcriptional activity of 5'-promoter of CERK. Truncation and mutation study suggests that ATRA-responsible region was mainly located in the tandem retinoic acid responsive elements (RARE) between -40 bp and the first exon. The electrophoresis mobility shift assay revealed that ATRA produced two retarded bands, which were erased by antibody against chicken ovalbumin upstream promoter transcription factor I (COUP-TFI), RARalpha, and RXRalpha, respectively. DNA pull down assay confirmed increased binding of these transcription factors to RARE. Transient expression of RAR, RXR, and COUP-TFI and siRNA transfection of these genes revealed that COUP-TFI inhibited CERK mRNA. Furthermore, chromatin immunoprecipitation assay showed the recruitment of co-repressors as well as three transcription factors. These results suggest that COUP-TFI was the ATRA responsive suppressive transcription factor of CERK gene transcription. PMID- 19903245 TI - Expression of chondromodulin-1 in the temporomandibular joint condylar cartilage and disc. AB - BACKGROUND: The temporomandibular joint (TMJ) cartilage consists of condylar cartilage and disc and undergoes continuous remodeling throughout post-natal life. To maintain the integrity of the TMJ cartilage, anti-angiogenic factors play an important role during the remodeling process. In this study, we investigated the expression of the anti-angiogenic factor, chondromodulin-1 (ChM 1), in TMJ cartilage and evaluate its potential role in TMJ remodeling. METHODS: Eight TMJ specimens were collected from six 4-month-old Japanese white rabbits. Safranin-O staining was performed to determine proteoglycan content. ChM-1 expression in TMJ condylar cartilage and disc was determined by immunohistochemistry. Three human perforated disc tissue samples were collected for investigation of ChM-1 and vascular endothelial growth factor (VEGF) distribution in perforated TMJ disc. RESULTS: Safranin-O stained weakly in TMJ compared with tibial articular and epiphyseal cartilage. In TMJ, ChM-1 was expressed in the proliferative and hypertrophic zone of condylar cartilage and chondrocyte-like cells in the disc. No expression of ChM-1 was observed in osteoblasts and subchondral bone. ChM-1 and VEGF were both similarly expressed in perforated disc tissues. CONCLUSIONS: ChM-1 may play a role in the regulation of TMJ remodeling by preventing blood vessel invasion of the cartilage, thereby maintaining condylar cartilage and disc integrity. PMID- 19903246 TI - Molecular markers in oral epithelial dysplasia: review. AB - The clinical and histologic features alone cannot accurately predict whether potentially malignant disorders of the oral mucosa remain stable, regress or progress to malignancy. Some of them, with or without epithelial dysplasia, may transform to invasive oral squamous cell carcinomas (OSCC). Identification of molecular markers which can predict disease progression is necessary to improve the management of these disorders. Many genes and signaling pathways have been shown to be involved in the development of OSCC. This review summarizes some molecular markers researched in the detection of pre-cancer. We highlight selected markers that are reported to be significantly associated with progression of potentially malignant disorders to OSCC. These include alterations in genes/pathways which control cellular signaling, cell cycle, apoptosis, genomic stability, cytoskeleton, angiogenesis, etc. However, these genetic tumor markers have so far not gained any use in routine diagnosis and their utility in the prediction of risk of malignant transformation remains unknown. It is, however, clear from the large number of studies, some described in this review, that multiple genes/pathways are involved in the progression from normal to metaplastic/dysplastic, and subsequently to cancer. It is therefore necessary to study those significant alterations in multiple genes simultaneously in biopsy samples from large cohorts of subjects. PMID- 19903247 TI - Anopheles hervyi in Niger: no evidence for a role in Plasmodium falciparum transmission. AB - Anopheles hervyi is an endemic mosquito species with a very limited spatial distribution in the south east of Niger. No new captures have been reported since the 1960s and its role in malaria transmission has not been studied. In the present study, the use of CDC light traps showed it to be much more abundant than previously found but there was no evidence to suggest it was a malaria vector in this region. The larval habitats have not been identified but the potential role of a saline lake in determining the distribution of this species is discussed. PMID- 19903248 TI - Children have rights, too. PMID- 19903249 TI - Australian Indigenous adolescents with chronic conditions: sociocultural context. AB - Scant information is available in the health literature on Australian Indigenous adolescents with chronic conditions and disabilities. Little is known about how Indigenous adolescence differs from mainstream adolescence, or how Indigenous adolescents manage chronic conditions. Health services are encouraged to engage in information sharing with Indigenous clients and to develop a collaborative approach to chronic condition management as a way to improve outcomes. PMID- 19903250 TI - UK health professionals' attitudes and knowledge regarding Human Papillomavirus (HPV) vaccination: a West Yorkshire Study. AB - AIM: To investigate the willingness of clinicians to recommend human papillomavirus (HPV) vaccination, the strength of support for a national HPV vaccine programme and to determine which factors, if any, affected these. METHODS: An online, invitation-only questionnaire was developed and distributed to three medical professional groups in the West Yorkshire Region, United Kingdom. RESULTS: Two hundred twenty-two responders were included in the final analysis, from the following specialties: general practice (62), paediatrics (103) and obstetrics and gynaecology (57). The majority of doctors were in favour of an National Health Service-funded national vaccination programme. Over 90% supported vaccination of girls as early as ages 11-13. Fewer doctors felt comfortable recommending vaccination to parents of girls under 16 than to young women. Latent class analysis demonstrated that doctors' self-rated knowledge of the HPV vaccine was an important determinant of willingness to recommend vaccination. Younger, more recently qualified doctors were less likely to be willing to recommend vaccination. CONCLUSIONS: There is widespread support for vaccination. Information provision to doctors will be important in maximising clinician confidence in recommending vaccination, and may be most beneficial when targeted at more junior doctors. PMID- 19903251 TI - Quality of care at the end of life in children with cancer. AB - AIM: Current Australian guidelines for the provision of paediatric palliative care highlight the importance of services being focused on the needs of the child and family. We aimed to establish parents' level of satisfaction with the quality of care currently being provided to children dying of cancer. METHODS: We interviewed 96 parents of children who died of cancer in Melbourne, Australia between 1996 and 2004 to ascertain how they rated the care provided to their child during the end-of-life period. RESULTS: A majority of parents were satisfied with the care provided by their primary oncologist, local doctors, palliative care services and home-care nurses. Most parents felt that discussions about key medical and treatment decisions were appropriate and clearly understood. Parents were generally satisfied with the leadership roles undertaken in decision-making in the end-of-life period; however, parents who were not satisfied indicated that they would like additional involvement of their primary oncologist. CONCLUSIONS: Current approaches to end-of-life care in children with cancer appear to be satisfactory. The main focus should continue to be on open and honest communication. PMID- 19903252 TI - A refugee with vomiting and weight loss. PMID- 19903253 TI - A case of symptomatic cholelithiasis in a 10-year-old girl. PMID- 19903254 TI - Cardiac disease causing death in a child with Wegener's granulomatosis. PMID- 19903255 TI - Streptococcus pneumoniae: uncommon cause of fatal neonatal sepsis. PMID- 19903257 TI - The Auckland Telepaediatrics Programme. PMID- 19903258 TI - Protective effects of affinity-purified antibody and truncated vaccines against Pseudomonas aeruginosa V-antigen in neutropenic mice. AB - Virulent P. aeruginosa strains express PcrV, one of the translocational components of the type III secretion system. PcrV has been reported to be a protective antigen against lethal P. aeruginosa infection. The PcrV region, which contributes to protective immunity against P. aeruginosa infection, was investigated by using genetically engineered, truncated PcrV proteins and affinity-purified anti-PcrV antibodies against the truncated PcrV proteins. The efficacy of active and passive immunization against PcrV was tested in mice with cyclophosphamide-induced immunosuppression by intraabdominal challenge of P. aeruginosa. Active immunization with either full-length PcrV1-294 or PcrV139-294 significantly improved the survival of mice infected with P. aeruginosa, while PcrV139-258, PcrV139-234, PcrV197-294, and PcrV261-294 were not protective. These results suggest that an effective PcrV vaccine needs to contain not only the Mab166 epitope (PcrV144-257) but also the carboxyl terminal tail of PcrV. In the case of passive immunization, administration of affinity-purified anti-PcrV IgG against either PcrV1-294 or PcrV139-258 showed significantly higher efficacy against lethal P. aeruginosa infection than did original anti-PcrV IgG and Mab166. The increased efficacy of affinity-purified anti-PcrV IgG implies that more potent anti-PcrV strategies are possible. The results of this study are crucial to the development of an effective PcrV vaccine for active immunization and to an appropriate blocking anti-PcrV antibody against P. aeruginosa infection in humans. PMID- 19903259 TI - Characterization of integrons and antimicrobial resistance genes in clinical isolates of Gram-negative bacteria from Palestinian hospitals. AB - Sixty Gram-negative bacterial isolates were collected from Palestinian hospitals in 2006. Thirty-two (53.3%) isolates showed multidrug resistance phenotypes. PCR and DNA sequencing were used to characterize integrons and antimicrobial resistance genes. PCR screening showed that 19 (31.7%) and five (8.3%) isolates were positive for class 1 and class 2 integrons, respectively. DNA-sequencing results for the captured antimicrobial resistance gene cassettes within class 1 integrons identified the following genes: dihydrofolate reductases, dfrA1, dfrA5, dfrA7, dfrA12, dfrA17 and dfrA25; aminoglycoside adenyltransferases, aadA1, aadA2, aadA5, aadA12 and aadB; aminoglycoside acetyltransferase, aac(6')-Ib; and chloramphenicol resistance gene, cmlA1. ESBL were identified in 25 (41.7%) isolates. The identified ESBL were bla(CTX-M-15), bla(CTX-M-56), bla(OXA-1), bla(SHV-1), bla(SHV-12), bla(SHV-32) and bla(TEM-1) genes. Moreover, we characterized the plasmid-mediated quinolone resistance genes, aac(6')-Ib-cr and qnrB2, which were detected in seven (11.7%) and two (3.3%) isolates, respectively. In this study various types of antibiotic resistance genes have been identified in Gram-negative bacteria from Palestinian hospitals, many of which are reported in the Middle East area for the first time. PMID- 19903260 TI - Study of human metapneumovirus-associated lower respiratory tract infections in Egyptian adults. AB - There is a deficiency in the data concerning the role of hMPV in lower respiratory tract infections in adults, and until now there has been no data available regarding the prevalence of hMPV in adults in our region. In the present study the association of hMPV with varieties of lower respiratory tract disorders in immunocompetent adult patients, either alone or with bacterial pathogens, has been highlighted. Eighty-eight patients were included in the study. They included 46 males and 42 females with an age range of 38-65 years. Patients presented with lower respiratory tract infections associated with acute exacerbation of asthma (67%), pneumonia (17%), and acute exacerbation of chronic obstructive lung diseases. Sputum and nasopharyngeal samples were obtained from the patients and subjected to a full microbiological study. In addition, detection of hMPV was performed by nested reverse transcriptase polymerase chain reaction. The pathogens isolated were Streptococcus pneumoniae 46.6%, Staphylococci aureus 35.2%, and human metapneumovirus 13.6%. Influenza virus and rhinovirus were each isolated from 4.5% of patients. Human metapneumovirus was associated with S. pneumoniae in 4.5% in studied patients, while in 9.1% it was the only pathogen found in those patients. The commonest clinical condition with significant association with human metapneumovirus was pneumonia. The clinical and laboratory studies demonstrated an association between lower respiratory tract infections in adults and hMPV either as sole pathogen or in association with Streptococcus pneumoniae. It was a common pathogen in community-acquired pneumonia. PMID- 19903261 TI - Covalent bonded Gag multimers in human immunodeficiency virus type-1 particles. AB - The oligomerization of HIV-1 Gag and Gag-Pol proteins, which are assembled at the plasma membrane, leads to viral budding. The budding generally places the viral components under non-reducing conditions. Here the effects of non-reducing conditions on Gag structures and viral RNA protection were examined. Using different reducing conditions and SDS-PAGE, it was shown that oligomerized Gag possesses intermolecular covalent bonds under non-reducing conditions. In addition, it was demonstrated that the mature viral core contains a large amount of covalent bonded Gag multimers, as does the immature core. Viral genomic RNA becomes sensitive to ribonuclease in reducing conditions. These results suggest that, under non-reducing conditions, covalent bonded Gag multimers are formed within the viral particles and play a role in protection of the viral genome. PMID- 19903262 TI - Archetype JC virus efficiently propagates in kidney-derived cells stably expressing HIV-1 Tat. AB - Pathogenic JCV with rearranged regulatory regions (PML-type) causes PML, a demyelinating disease, in the brains of immunocompromised patients. On the other hand, archetype JCV persistently infecting the kidney is thought to be converted to PML-type virus during JCV replication in the infected host under immunosuppressed conditions. In addition, Tat protein, encoded by HIV-1, markedly enhances the expression of a reporter gene under control of the JCV late promoter. In order to examine the influence of Tat on JCV propagation, we used kidney-derived COS-7 cells, which only permit archetype JCV, and established COS tat cells, which express HIV-1 Tat stably. We found that the extent of archetype JCV propagation in COS-tat cells is significantly greater than in COS-7 cells. On the other hand, COS-7 cells express SV40 T antigen, which is a strong stimulator of archetype JCV replication. The expression of SV40 T antigen was enhanced by HIV-1 Tat slightly according to real-time RT-PCR, this was not closely related to JCV replication in COS-tat cells. The efficiency of JCV propagation depended on the extent of expression of functional Tat. To our knowledge, this is the first report of increased production of archetype JCV in a culture system using cell lines stably expressing HIV-1 Tat. We propose here that COS-tat cells are a useful tool for studying the role of Tat in archetype JCV replication in the development of PML. PMID- 19903263 TI - N-linked glycan-dependent interaction of CD63 with CXCR4 at the Golgi apparatus induces downregulation of CXCR4. AB - Efficient downregulation of CXCR4 cell surface expression by introduction of the CD63 gene has previously been reported by us. In the present study, it was found that CD63 and its mutant efficiently interact with CXCR4 in live cells and that CD63-induced downregulation and interaction are significantly abrogated by the N linked glycosylation inhibitor, TM. Furthermore, the downregulation and interaction were clearly attenuated by alternation of all three N-linked glycosylation sites in CD63. Either CD63 or CD63DeltaN formed a complex with CXCR4 at the Golgi apparatus and the late endosomes, while CD63 GD mutants lost the ability to form a complex with CXCR4 exclusively at the Golgi apparatus. These findings suggest that CD63 interacts with CXCR4 through the N-linked glycans-portion of the CD63 protein and that the complex induces direction of CXCR4 trafficking to the endosomes/lysosomes, rather than to the plasma membrane. At the Golgi apparatus, there may be lysosome protein (CD63)-associated machinery that influences trafficking of other membrane proteins. PMID- 19903264 TI - Enhanced immune response to pneumococcal infection in malnourished mice nasally treated with heat-killed Lactobacillus casei. AB - The present study analyzed whether nasal administration of viable and non-viable Lactobacillus casei CRL 431 to immunocompromised mice was capable of increasing resistance against Streptococcus pneumoniae. Weaned mice were malnourished after consuming a PFD for 21 days. Malnourished mice were fed a BCD for 7 days or BCD for 7 days with viable or non-viable L. casei nasal treatments on day 6 and day 7 (BCD+LcV and BCD+LcN, respectively). The MNC group received PFD whereas the WNC mice consumed BCD. MNC mice showed greater lung colonization, more severe lung injuries, impaired leukocyte recruitment and reduced antibodies and cytokine production when compared with WNC mice. Administration of L. casei increased the resistance of malnourished mice to the infection. Both BCD+LcV and BCD+LcN treatments prevented the dissemination of the pathogen to the blood and induced its lung clearance. BCD+LcV or BCD+LcN groups showed improved production of TNF alpha and activity of phagocytes in the respiratory tract, an effect that was not observed in the BCD control group. In addition, IL-4 and IL-10 were significantly increased in BCD+LcV and BCD+LcN groups, which correlated with the increase in the levels of specific respiratory IgA. The nasal treatments with L. casei were also effective at stimulating the production of specific IgG at both the systemic and the respiratory levels. The comparative study between the viable and the non viable bacteria demonstrated that viability would be an important factor to achieve maximum protective effects. However, the results from this study suggest that heat-killed lactic acid bacteria are also effective in the immunomodulation of the systemic and respiratory immune system. PMID- 19903265 TI - Altered profiles of intestinal microbiota and organic acids may be the origin of symptoms in irritable bowel syndrome. AB - BACKGROUND: The profile of intestinal organic acids in irritable bowel syndrome (IBS) and its correlation with gastrointestinal (GI) symptoms are not clear. We hypothesized in this study that altered GI microbiota contribute to IBS symptoms through increased levels of organic acids. METHODS: Subjects were 26 IBS patients and 26 age- and sex-matched controls. Fecal samples were collected for microbiota analysis using quantitative real-time polymerase chain reaction and culture methods, and the determination of organic acid levels using high-performance liquid chromatography. Abdominal gas was quantified by image analyses of abdominal X-ray films. Subjects completed a questionnaire for GI symptoms, quality of life (QOL) and negative emotion. KEY RESULTS: Irritable bowel syndrome patients showed significantly higher counts of Veillonella (P = 0.046) and Lactobacillus (P = 0.031) than controls. They also expressed significantly higher levels of acetic acid (P = 0.049), propionic acid (P = 0.025) and total organic acids (P = 0.014) than controls. The quantity of bowel gas was not significantly different between controls and IBS patients. Finally, IBS patients with high acetic acid or propionic acid levels presented with significantly worse GI symptoms, QOL and negative emotions than those with low acetic acid or propionic acid levels or controls. CONCLUSIONS & INFERENCES: These results support the hypothesis that both fecal microbiota and organic acids are altered in IBS patients. A combination of Veillonella and Lactobacillus is known to produce acetic and propionic acid. High levels of acetic and propionic acid may associate with abdominal symptoms, impaired QOL and negative emotions in IBS. PMID- 19903266 TI - Pseudo-ventricular over and under-sensing during an episode of double tachycardia. What is the mechanism? PMID- 19903267 TI - Heart rate recovery and chronotropic incompetence in patients with subclinical hypothyroidism. AB - BACKGROUND: Heart rate recovery (HRR) and chronotropic incompetence (CI) in patients with subclinical hypothyroidism (SCH) has not been explored previously. The aim of the present study was to evaluate the HRR and CI in patients with SCH. METHODS: Twenty-five patients (11 men, 14 women with a mean age of 36 + or - 10 years) who were diagnosed SCH determined by an increased serum thyrothrophine (TSH) concentration (>4.0 ng/mL) and the normal free triiodothyronine (fT3) and free thyroxin (fT4) levels, were included in the study. The control group of healthy individuals with normal TSH (12 males, 15 females) with a mean age of 36 + or - 3 years was also included. Two groups were well matched for age, sex, and body mass index. Medical history, physical examination, electrocardiogram, treadmill exercise testing, and chest radiogram were performed for all participants. RESULTS: The characteristics of SCH patients and control cases were similar with regard to age, sex, and BMI except for TSH levels. Serum TSH levels were significantly higher in SCH patients than the controls (P < 0.001). No significant differences were observed in the changes of heart rate (HR), exercise tolerance (metabolic equivalents), or systolic and diastolic blood pressures at rest or during exercise between the groups, whereas HRR and CI were significantly lower during exercise testing in the SCH patients compared to controls (P < 0.003; P < 0.03, respectively). CONCLUSION: The results of the present study demonstrated that SCH can cause impaired cardiovascular autonomic function and attenuated HR response to exercise. (PACE 2010; 2-5). PMID- 19903268 TI - Education at the margins and beyond borders. PMID- 19903269 TI - Relational aggression and adverse psychosocial and physical health symptoms among urban adolescents. AB - OBJECTIVES: The purpose of this study was to examine relational aggression and its relationship with adverse psychosocial and physical health symptoms among urban, African American youth. DESIGN AND SAMPLE: Quantitative, cross-sectional survey design. The sample consisted of 185 predominantly African American (95.1%) seventh-grade students (mean age: 13.0; female: 58%) attending 4 urban middle schools. MEASURES: The Children's Social Behavior Scale and Social Experience Questionnaire were used to measure relational aggression and relational victimization. The Pediatric Symptom Checklist was used to assess psychosocial difficulties, including internalizing behaviors, externalizing behaviors, and attention problems. Physical health symptoms were measured with questions about colds/flu, headaches, and stomach aches. RESULTS: 2-way multivariate analysis of variance revealed significant differences in externalizing behavior, with perpetrators reporting higher levels than nonperpetrators. Victims reported more internalizing behavior than nonvictims; however, this was only significant for males. For females, significant negative effects on health outcomes were found, resulting from the interaction of perpetration and victimization. CONCLUSIONS: Findings suggest that relational aggression is a common occurrence among urban, minority adolescents and may result in adverse health outcomes. These results provide several avenues for future research and implications for healthcare practice. Intervention strategies are needed to prevent relational aggression and continual or subsequent adverse health symptoms. PMID- 19903270 TI - Cardiometabolic health of Chinese older adults with diabetes living in Beijing, China. AB - OBJECTIVES: Using the PRECEDE-PROCEED model, this study examined the presence of metabolic syndrome and modifiable cardiovascular disease (CVD) risk factors associated with metabolic syndrome among Chinese adults with diabetes living in Beijing, China. DESIGN AND SAMPLE: The cross-sectional study collected data through face-to-face interviews. The study included 73 Chinese older adults with diabetes. Their mean age was 68 years (+ or - 7.66), with a range from 52 to 90 years. MEASUREMENTS: Data were collected on demographic characteristics, blood pressure (BP), body mass index (BMI), waist circumference, lipid profile and fasting glucose, physical activity, diet, and health status. RESULTS: The great majority (85%) had metabolic syndrome; 65% had hypertension; 52% had high levels of low-density lipoproteins, and 80.6% had a high level of fasting glucose. Half of the participants (51.4%) were overweight, 16.7% were obese, and 86.3% had central obesity. Age, gender, BMI, income, insurance, smoking history, physical activity, and diet explained 23% of the variance in the metabolic syndrome component, systolic blood pressure. CONCLUSIONS: The association of predisposing and enabling factors and health behavior with the metabolic syndrome needs to be further explored. Persons with diabetes should have regular health screenings to check for blood pressure, BMI, cholesterol, glucose, and triglycerides in order to decrease the risks associated with metabolic syndrome and CVD. PMID- 19903271 TI - Antenatal care utilization in a conflict-affected district of Northern Sri Lanka. AB - OBJECTIVES: To assess utilization of antenatal care (ANC) and its determinants in a conflict-affected area in Northern Sri Lanka. DESIGN AND SAMPLE: A cross sectional study was conducted among pregnant mothers who had completed 36 weeks of gestation, and were admitted or referred to 2 leading hospitals in Vavuniya district. MEASURES: Data were collected from 392 mothers using an interviewer administered questionnaire. RESULTS: 55% of the respondents reported that their families were affected directly by conflict. Approximately 68% of mothers were registered by public health midwife (PHM) for ANC; 31.4% were registered before 12 weeks of gestation; 38.5% were visited at home by PHM; 37.8% had the first clinic visit before 12 weeks; and 90.1% made at least 4 clinic visits. The average number of antenatal visits to specialist clinics (5.34, 95%CI: 5.03-5.65) was significantly higher than field clinics (2.97, 95%CI: 2.69-3.25). On average, a pregnant woman was seen by PHM at home 1.47 times. ANC utilization was significantly poor in women whose families were affected by conflict, living in active conflict areas and urban regions, had a lower education level, and were not involved in decision making on ANC. CONCLUSION: Provision of field antenatal clinics, early entry to ANC, and domiciliary care by PHM need improvement. PMID- 19903273 TI - Depression and language acculturation correlate with smoking among older Asian American adolescents in New York City. AB - OBJECTIVE: To explore the correlates of smoking status among a sample of Asian American adolescents in New York City (NYC). DESIGN AND SAMPLE: This descriptive, correlational study compared current smokers and current nonsmokers in terms of age, gender, ethnicity, academic performance, acculturation, depressive symptoms, smoking history, and parental, sibling, and peer smoking. The convenience sample included 328 Asian American adolescents, ages 16-19, who lived in NYC, and were recruited from members, friends, and affiliates of 6 organizational sources. MEASURES: The study used demographic, depression (Center for Epidemiological Studies Depression scale), English language acculturation, and smoking opportunity survey questionnaires. RESULTS: Statistical analyses revealed that being older, being more English-language acculturated, having poor academic performance, increased depressive symptoms, and having siblings and peers who smoked were significantly associated with current smoking. CONCLUSIONS: Asian American adolescents initiate smoking later than other groups. Depressive symptoms were at high levels in the sample and were correlated with smoking. Thus, for Asian American adolescents, smoking prevention programs should be extended to later grades, with particular attention to low-performing students and those who are more acculturated. Likewise, programs that identify and assist adolescents with depressive symptoms may be useful in smoking prevention. PMID- 19903272 TI - Effects of exercise program on physical fitness, depression, and self-efficacy of low-income elderly women in South Korea. AB - OBJECTIVE: This study aimed to examine the effects of exercise programs on physical fitness, depression, and self-efficacy in low-income elderly women (age > or = 75). DESIGN AND SAMPLE: A pretest-posttest experimental research design with a control group was used. The sample consisted of 26 women in the exercise group and 22 women in the wait-list control group in Seoul, Korea. MEASURES: The measures of physical fitness included body mass index, cardiopulmonary endurance (blood pressure and heart rate), muscle strength (hand grip strength), flexibility (degree of bending of the upper body), and balance (duration of time for which the subject could stand on one foot). Depression and self-efficacy were measured by the Center for Epidemiological Studies Depression scale and a modified form of Lee's scale, respectively. All measures were obtained twice: at baseline and at the completion of the exercise program. INTERVENTION: The exercise program consisted of 4 weeks of education along with 8 weeks of physical exercise. RESULTS: After the intervention, significant improvements were found in depression, self-efficacy, and all measures of physical fitness, except heart rate and flexibility, in the experimental group. CONCLUSION: The exercise program may be recommended as a method to maintain and promote the health of low-income elderly women. PMID- 19903274 TI - A grounded theory study of action/interaction strategies used when Taiwanese families provide care for formerly suicidal patients. AB - OBJECTIVES: There is a dearth of evidence on the care that families provide to their relatives after they have been discharged from hospital following an attempted suicide. The aim of this study was to explore ex-patients' and family caregivers' perceptions of the care provided at home following hospital discharge. DESIGN AND SAMPLE: A qualitative approach using Grounded Theory was adopted. Suicidal ex-patients (n=15) and family caregivers (n=15) were contacted in the south of Taiwan. MEASURES: Data were collected through interviews and the data were analyzed using open, axial, and selective coding. RESULTS: A substantive theory was developed from the findings. However, for the purpose of this paper, the section of the paradigm model named the action/interaction strategies is presented and discussed. Three categories emerged in the action/interaction section relating to the family care of relatives who had been suicidal. They were: (1) "guarding the person day and night," which helped to ensure that their relatives felt safe; (2) "maintaining the activities of daily living," which promoted their physical health and recovery; and (3) "creating a nurturing environment," which facilitated their mental and emotional healing. CONCLUSIONS: Public health nurses could use the findings of this study as a theoretical map when providing health information to family caregivers during home visits. PMID- 19903275 TI - Population health surveillance practice of public health nurses. AB - OBJECTIVE: To report the population health surveillance functions of public health nurses and to describe factors that impede these functions. DESIGN AND SAMPLE: An interpretive qualitative study was conducted in Public Health Service areas in Eastern Canada. Participants were public health nurses (n=55) with an average of 14.5 years of pertinent work experience. MEASURES: Semistructured face to-face, telephone interviews, and focus groups were conducted, transcribed, coded, and analyzed. RESULTS: The nurses in this study used ecosocial population health surveillance functions that included multilevel societal influences on health. Extensive interprofessional and intersectoral networks were foundational to their surveillance work, allowing them to monitor what was occurring in the community and transfer this knowledge into various systems to contribute toward improved health outcomes. However, the nurses did not acknowledge the significance of their population health surveillance work, and documentation structures did not support these surveillance functions. CONCLUSION: New surveillance methods and documentation structures that reflect an ecosocial surveillance approach are needed that are more consistent with public health nurses' population-focused practice. PMID- 19903276 TI - Nataniel, NAFTA, and Public Health at the U.S.-Mexico Border. AB - Advocating overall improvements in health for individuals and communities is a daunting but important task for nurses in particular, and for health care professionals in general. This is particularly true when focusing on the population along the 2,000-mile U.S.-Mexico border, a unique region in which distinct cultures, economies, and political systems meet. The purpose of this paper is to confront the assumption that trade and economic expansion automatically translate into improved public health, and to explore policy implications of the public health situation at the border. It uses a meta narrative, an overarching story that draws on and illustrates collective stories from 300 participants in a study of mental health disparities, to argue for a more nuanced and complex understanding of health among the largely Hispanic population in this region. PMID- 19903277 TI - The San Francisco collaborative: an evaluation of a partnership between three schools of nursing and a public health department. AB - Many factors are contributing to a decline in the number of nurses who opt to choose public health nursing as a career option. One factor is the lack of preceptors in public health, which has led to the placement of nursing students in nontraditional clinical settings. Thus, many nursing students are not exposed to public health nursing while still in school. Graduating students may not have a clear idea of what a public health nurse is or does. The purpose of the study was to describe a collaborative project between three schools of nursing aimed at increasing interest in public health nursing among undergraduate nursing students. The study method involved analysis of student, faculty, and staff feedback received after an orientation to public health nursing. Nursing students found the experience valuable. Participation in the collaborative project has increased clinical experiences for nursing students in public health, increased exposure of nursing students to public health nursing, and has led to opportunities for both students and faculty within an urban public health department. PMID- 19903278 TI - Psychometric assessment of the Brazilian version of the Breastfeeding Self Efficacy Scale. AB - OBJECTIVES: The purpose of this study was to translate and psychometrically assess the Breastfeeding Self-Efficacy Scale (BSES) among women living in Fortaleza, CE, Brazil, and examine the relationship between breastfeeding self efficacy (BSE) and maternal demographic variables. DESIGN AND SAMPLE: This methodological study is the first translation of BSES conducted in South America. The psychometric assessment of the original study was replicated. This methodological study enrolled a group (sample of judgment) of 117 pregnant women. MEASURES: BSES and maternal sociodemographic variables were studied. RESULTS: The Cronbach's alpha coefficient for the translated BSES was .88. Significant differences in BSES scores were found among mothers with a previous satisfactory breastfeeding experience (M = 145.81; SD = + or - 6.82, p = .0001). Significant relationships were found among prenatal BSE and maternal age (r = .228; p = .01), educational level (r = .234; p = .01), and marital status (r = .183; p = .04). No relationship was found among BSE and maternal occupation, family income, or number of pregnancies. CONCLUSIONS: Altogether, our findings suggest that BSES translated into Portuguese may be a reliable and valid measure to assess maternal BSE in Brazilian culture. Minor changes may be needed to use it in other Portuguese-speaking countries, such as Portugal and Mozambique. PMID- 19903280 TI - Should total platelet mass be a criterion for prophylactic platelet transfusion? PMID- 19903281 TI - Blood salvage and cancer surgery: should we do it? PMID- 19903282 TI - Young donors: opportunity, responsibility, vulnerability--report on YDAR task force. PMID- 19903283 TI - Transfusion medicine illustrated. Thrombus formation during platelet donation. PMID- 19903284 TI - Age-related donor return patterns among first-time blood donors in the United States. AB - BACKGROUND: Committed repeat donors are vital to the continued success of blood collections, yet the effect of age of first-time (FT) donation on return behavior is poorly described. Sixteen-year-old donors are increasingly allowed to donate and have the highest rates of adverse events, which negatively impacts return behavior. STUDY DESIGN AND METHODS: Annual cohorts of allogeneic FT donors from 2005 and 2006 were selected within the American Red Cross system and followed for 25 and 13 months, respectively. Return and total yield rates among different age groups were compared. RESULTS: A total of 2.3 million FT donors from 2005 and 2006 gave 4.2 million donations during the study. Sixteen- to 19-year old FT donors made up 41% of the FT donor base in 2005 and 16-, 17-, 18-, and 19-year olds, respectively, had initial return rates of 62, 52, 35, and 28% and yield rates of 2.0, 1.76, 1.51, and 1.41 over 13 months. Multivariate analysis of FT yield rates shows that younger (16 and 17 years) and older (50+ years) donors, males, blood group O donors, and those without any initial adverse reaction are most likely to return. Increasing severity of donor adverse reactions correlated with a reduction in yield and return rates. CONCLUSION: FT 16-year-old donors had the highest return and yield rates despite the negative impact of increased adverse event rates. Donation at young age is critical to building a cadre of committed repeat donors but donor reactions must be addressed to ensure the donors' well-being and to sustain return behavior. PMID- 19903285 TI - A new strategy to improve the cost-effectiveness of human immunodeficiency virus, hepatitis B virus, hepatitis C virus, and syphilis testing of blood donations in sub-Saharan Africa: a pilot study in Burkina Faso. AB - BACKGROUND: In Africa where blood-borne agents are highly prevalent, cheaper and feasible alternative strategies for blood donations testing are specifically required. STUDY DESIGN AND METHODS: From May to August 2002, 500 blood donations from Burkina Faso were tested for hepatitis B surface antigen (HBsAg), human immunodeficiency virus (HIV), syphilis, and hepatitis C virus (HCV) according to two distinct strategies. The first strategy was a conventional simultaneous screening of these four blood-borne infectious agents on each blood donation by using single-marker assays. The second strategy was a sequential screening starting by HBsAg. HBsAg-nonreactive blood donations were then further tested for HIV. If nonreactive, they were further tested for syphilis. If nonreactive, they were finally assessed for HCV antibodies. The accuracy and cost-effectiveness of the two strategies were compared. RESULTS: By using the simultaneous strategy, the seroprevalences of HBsAg, HIV, syphilis, and HCV among blood donors in Ouagadougou were estimated to be 19.2, 9.8, 1.6, and 5.2%. No significant difference of HIV, syphilis, and HCV prevalence rates was observed by using the sequential strategy (9.2, 1.9, and 4.7%, respectively). Whatever the strategy used, 157 blood donations (31.4%) were found to be reactive for at least one transfusion-transmissible agent and were thus discarded. The sequential strategy allowed a cost decrease of euro 908.6, compared to the simultaneous strategy. Given that approximately there are 50,000 blood donations annually in Burkina Faso, the money savings reached potentially euro 90,860. CONCLUSIONS: In resource limited settings, the implementation of a sequential strategy appears as a pragmatic solution to promote safe blood supply and ensure sustainability of the system. PMID- 19903286 TI - Hepatitis C virus (HCV) core antigen detection in HCV RNA-positive/anti-HCV negative Polish blood donors identified by nucleic acid testing. PMID- 19903288 TI - Transfusion medicine illustrated: beware of fibrin residues. PMID- 19903289 TI - Platelet quality measured with dynamic light scattering correlates with transfusion outcome in hematologic malignancies. AB - BACKGROUND: A clinically meaningful test for platelet (PLT) quality could improve the transfusion management of patients. The aim of this pilot study was to determine whether a new measure of PLT quality and function based on dynamic light scattering (DLS) correlates with transfusion outcome. STUDY DESIGN AND METHODS: For a total of 160 transfusions, the pretransfusion, 1 hour posttransfusion, and 24-hour posttransfusion PLT counts were routinely measured in 49 patients (31 male, 18 female; age 46 +/- 15 years) with hematologic malignancies. The corrected count increments (CCIs) at 1 hour (PLT recovery) and 24 hours (PLT survival) were calculated and used as the transfusion outcome measures. The ThromboLUX score (LightIntegra Technology, Inc., Vancouver, BC, Canada; range, 0-40; cutoff, 12) and the PLT morphology score of the PLT concentrates were determined and compared to transfusion outcome. RESULTS: The CCIs and ThromboLUX scores were normally distributed and showed a strong correlation (n = 96, in the mixed regression model the adjusted coefficient is R = 0.6292, p < 0.0001), while other variables such as product type, age, and microscopic PLT morphology score were not correlated with transfusion outcome (p > 0.05). Importantly, 12 of 96 transfusions with poor PLT quality were clinically ineffective, that is, did not adequately increase the PLT counts in the recipients. One patient died after receiving three consecutive ineffective PLT transfusions with a low ThromboLUX score. CONCLUSION: In this pilot study, the ThromboLUX score strongly correlated with transfusion outcome (PLT recovery and survival) independent of clinical and product issues. PMID- 19903290 TI - Risk for malaria in United States donors deferred for travel to malaria-endemic areas. AB - BACKGROUND: Deferral for travel to malaria-endemic areas excludes many blood donors in the United States. Most transfusion-transmitted malaria is associated with lengthy residence in malaria-endemic areas rather than routine travel. This study compares the impact of existing deferral requirements to the risk that a presenting donor with malaria travel history harbors malaria parasites under current and hypothetical alternate regulations. STUDY DESIGN AND METHODS: Deferred donors from six blood centers were sampled to estimate a national cohort of donors deferred annually for malaria travel to different geographic regions. Risk for malaria infection after travel to each region and distribution of incubation periods for each malaria species were estimated for US travelers. Region-specific travel risks were used to estimate the risk that a presenting blood donor with malaria travel might asymptomatically harbor malaria parasites at different intervals after return to the United States. RESULTS: Travel to Africa presents risk for malaria infection greater than 1000 times that of travel to malaria-endemic parts of Mexico, yet Mexico accounts for more than 10 times as many deferred donors. Shortening the deferral period from 12 to 3 months for travelers to Mexico increases the risk of collecting a contaminated unit by only 1 unit per 57 years (sensitivity analysis, 1 every 29-114 years), at annual gain of more than 56,000 donations. CONCLUSION: This study provides the first systematic appraisal of the US requirements for donor qualification regarding travel to malarial areas. Consideration should be given to relaxing the guidelines for travel to very-low-risk areas such as Mexico. PMID- 19903291 TI - Estimation of sensitivity and specificity of several Trypanosoma cruzi antibody assays in blood donors in Argentina. AB - BACKGROUND: The absence of a gold standard test for Trypanosoma cruzi antibodies represents a problem not only for the evaluation of screening tests, but also for appropriate blood donor counseling. The aim of this study was to estimate the sensitivity and specificity of multiple blood donor screening tests for T. cruzi antibodies in Argentina. STUDY DESIGN AND METHODS: From June 2006 to March 2007 a sample of 1455 blood donors was recruited from two blood banks in Chaco province, an area of Argentina with highly endemic T. cruzi infection. Samples were tested by three epimastigote lysate enzyme immunoassays (EIAs), one recombinant antigen EIA, two indirect hemagglutination assay (IHA) tests, a particle agglutination assay (PA), and a research trans-sialidase inhibition assay (TIA). Sensitivity and specificity were estimated using latent class analysis (LCA). RESULTS: LCA estimated the consensus prevalence of T. cruzi infection at 24.5%. Interassay correlation was higher among the four EIA tests and TIA compared to IHA tests. Assay sensitivities varied from 96 to 99.7 for different EIAs, 91% for TIA, 84% for PA, and 66 to 74% for IHA tests. Relative to the LCA, assay specificities were from 96% to almost 100%. CONCLUSION: Based on the comparison of several tests in a large population from an endemic area for T. cruzi infection, our data showed an adequate sensitivity for EIA tests in contrast to PA and IHA assays. The latter tests should no longer be used for blood donor screening. PMID- 19903292 TI - Mini buffy coat photopheresis for children and critically ill patients with extracorporeal photopheresis contraindications. AB - BACKGROUND: Conventional extracorporeal photopheresis (ECP) has proven efficacy for the treatment of several diseases but is limited to patients with sufficient body weight. A novel simplified mini buffy coat ECP technique that allows treatment of small children and patients with apheresis contraindications has been developed. STUDY DESIGN AND METHODS: White blood cell (WBC)-rich buffy coat fractions were prepared from 5 to 8 mL/kg whole blood in a closed system, diluted, and ultraviolet A (UVA)-irradiated after addition of 8-methoxypsoralen (8-MOP). Apoptosis and cell death were analyzed by annexin V and 7 aminoactinomycin staining. Lymphocyte proliferation was measured after CD3/CD28 and phytohemagglutinin (PHA) stimulation. Autologous residual blood and UVA irradiated buffy coat were returned to the patients. Fifty-six mini buffy coat ECP procedures were applied to three children with acute steroid-refractory skin graft-versus-host disease and apheresis contraindications. RESULTS: Mean whole blood and buffy coat volumes were 166 (+/-61.8) and 8 (+/-1.6) mL, respectively, and resulted in a hematocrit of 2.2% (+/-0.4) after saline dilution (median +/- SD). UVA irradiation of 8-MOP buffy coat preparations resulted in significant induction of WBC apoptosis at 48-72 hours (p 0.05). A median 76.5% of G-CSF-mobilized CD34+ HSCs coexpressed the CD133+ antigen (range, 23.1-97.9). CONCLUSIONS: A higher number of CD133+/CD34+ HSCs in the graft was not clearly associated with a shorter neutrophil or PLT recovery time in either allogeneic or autologous recipients. PMID- 19903295 TI - The role of the elution in antibody investigations. AB - BACKGROUND: The direct antiglobulin test (DAT) commonly detects immunoglobulin G (IgG) molecules or complement fragments on the red blood cell (RBC) surface. If IgG antibodies are present then elution procedures can be performed to identify the specificity of these antibodies. Our reference laboratory performs elutions on the RBCs of those patients who have received cellular blood products in the past 30 days and have either a newly identified positive DAT with anti-IgG or the agglutination strength is increased over a previous DAT and if ordered by a clinician regardless of transfusion history. This study questioned how frequently elutions contributed novel serologic information under our reference laboratory's current policy or whether elutions should be performed in more selective serologic conditions. STUDY DESIGN AND METHODS: Recipients whose RBCs underwent eluate testing were identified from the blood bank's database and information about the antecedent DAT and antibody detection test and eluate was recorded. RESULTS: In total 648 eluates were evaluated and 82 of 648 (12.7%) revealed a novel antibody not present in the serum (an informative eluate). In 2 of 82 informative eluates non-anti-A/B alloantibodies that were not present in the serum were detected: one example each of anti-D and anti-E. Both were associated with a microscopically positive antecedent DAT. The rate of an informative eluate was higher when the antibody detection test was negative. CONCLUSION: The strength of the DAT does not indicate the likelihood of an informative eluate. Performing an eluate when the antibody detection test is positive has limited value. PMID- 19903297 TI - Regulating blood manufacturing software: report of a conference. PMID- 19903296 TI - ABO-identical versus nonidentical platelet transfusion: a systematic review. AB - BACKGROUND: The significance of ABO matching for platelet (PLT) transfusion has not been clearly defined. The primary objective of this report is to assess whether ABO-identical PLT transfusion is associated with improved mortality and/or morbidity for patients with hematologic/oncologic disorders. STUDY DESIGN AND METHODS: A systematic review to January 2009 was conducted. Data on mortality, morbidity, PLT refractoriness, and PLT increment after transfusion were abstracted. RESULTS: A total of 100 citations were identified. Nineteen studies were included in the systematic review. A total of 1502 patients from three randomized controlled trials and 16 observational studies were included. Survival, bleeding events, and transfusion reactions were only considered as secondary outcomes in the reports reviewed. The PLT count increment was the primary outcome of several studies and was consistently higher with ABO-identical PLT transfusion. The largest difference in increment between ABO-identical and nonidentical PLT transfusion was 4 x 10(9)/L. No consistent benefit in clinical outcomes was noted. Survival was assessed in three reports with conflicting results. Although two studies described bleeding as an outcome, the assessment of hemorrhage was considered inadequate. In six studies, ABO-nonidentical PLT transfusion was not associated with transfusion reactions, and the results from four studies addressing the impact of ABO-identical PLT transfusion on PLT and red blood cell utilization were conflicting. CONCLUSION: ABO-identical PLT transfusion results in a higher PLT increment. Randomized controlled trials are required to definitely determine the effect of ABO-identical PLT transfusion on survival, bleeding events, or transfusion reactions. PMID- 19903298 TI - Hemoglobin-based oxygen carriers: current status and future directions. PMID- 19903299 TI - Screening for glucose-6-phosphate dehydrogenase deficiency in blood donors. PMID- 19903301 TI - Health-related quality of life in hospital inpatients with pressure ulceration: assessment using generic health-related quality of life measures. AB - The objective of this study was to determine the impact of pressure ulceration on health-related quality of life (HRQoL) and to undertake a pilot study for a future larger study. The study comprised two parts. First, data from a large UK prospective cohort study were analyzed and the HRQoL of 218 people with pressure ulcers was compared with that of 2,289 people without ulcers using the Short Form 36 (SF-36) questionnaire. After adjusting for age, sex, and comorbidities, patients with pressure ulceration had significantly lower scores for both the physical (coefficient=-3.12, p<0.001) and mental (coefficient=-1.50, p=0.04) component summary scores of the SF-36. Second, a small pilot study was conducted to explore use of other tools. HRQoL was assessed in six patients with and 16 patients without pressure ulcers using the SF-36, the EQ-5D and a pain visual analog scale. SF-36 scores indicated that patients with pressure ulcers had significantly poorer physical functioning (d=22.3, p=0.001), role limitations due to physical problems (d=12.9, p=0.02), and vitality (d=20.6, p=0.04) than those without. EQ-5D scores were also poorer for patients with pressure ulceration, for both the visual analog scale (d=19.2, p=0.02) and the index (d=0.29, p=0.08). Patients with pressure ulceration had more perceived pain than those without; however, this difference was of borderline significance (d=-23.9, p=0.06). Pressure ulceration therefore has an impact on HRQoL that is measurable and persists after adjusting for potential confounding. PMID- 19903300 TI - Human skin wounds: a major and snowballing threat to public health and the economy. AB - ABSTRACT In the United States, chronic wounds affect 6.5 million patients. An estimated excess of US$25 billion is spent annually on treatment of chronic wounds and the burden is rapidly growing due to increasing health care costs, an aging population and a sharp rise in the incidence of diabetes and obesity worldwide. The annual wound care products market is projected to reach $15.3 billion by 2010. Chronic wounds are rarely seen in individuals who are otherwise healthy. In fact, chronic wound patients frequently suffer from "highly branded" diseases such as diabetes and obesity. This seems to have overshadowed the significance of wounds per se as a major health problem. For example, NIH's Research Portfolio Online Reporting Tool (RePORT; http://report.nih.gov/), directed at providing access to estimates of funding for various disease conditions does list several rare diseases but does not list wounds. Forty million inpatient surgical procedures were performed in the United States in 2000, followed closely by 31.5 million outpatient surgeries. The need for post surgical wound care is sharply on the rise. Emergency wound care in an acute setting has major significance not only in a war setting but also in homeland preparedness against natural disasters as well as against terrorism attacks. An additional burden of wound healing is the problem of skin scarring, a $12 billion annual market. The immense economic and social impact of wounds in our society calls for allocation of a higher level of attention and resources to understand biological mechanisms underlying cutaneous wound complications. PMID- 19903302 TI - Modulation of scarring in a liquid environment in the Yorkshire pig. AB - ABSTRACT Decreased inflammatory response seen in wet wound healing may be correlated with diminished scarring. This study seeks to test this hypothesis and to validate a model of scarring in the Yorkshire pig. Four Yorkshire pigs were used to create 36 dorsal wounds per pig (144 wounds total) in the following groups: full-thickness excisional, partial thickness, meshed split-thickness skin grafts, sheet split-thickness skin grafts, minced skin, and incisional wounds. Wounds were randomized into wet and dry groups. Wet wounds were enclosed in polyurethane chambers with 2 mL of normal saline. Dry wounds were covered with regular gauze. Terminal biopsies were performed at 72 hours and day 28. Histology demonstrated significantly less inflammatory infiltrate, thicker neoepidermis, more pronounced rete ridge formation, and decreased scar tissue thickness in wet wounds. The mean macroscopic scar surface area was significantly decreased in full-thickness excisional wet wounds compared with dry wounds (61.2 mm(2) vs. 150.8 mm(2), p<0.01). Hydroxyproline content was decreased in full-thickness wet compared with dry groups (44.81 vs. 62.21 mg/g, p<0.01). Tensile strength was 90% greater in full-thickness wet compared with dry groups (p<0.01). Healing in the liquid environment significantly reduced scar formation. This model will allow for future investigation of high-concentration topical scar-modulating agents in the liquid environment. PMID- 19903303 TI - Development of a new chitosan hydrogel for wound dressing. AB - Wound healing is a complex process involving an integrated response by many different cell types and growth factors in order to achieve rapid restoration of skin architecture and function. The present study evaluated the applicability of a chitosan hydrogel (CH) as a wound dressing. Scanning electron microscopy analysis was used to characterize CH morphology. Fibroblast cells isolated from rat skin were used to assess the cytotoxicity of the hydrogel. CH was able to promote cell adhesion and proliferation. Cell viability studies showed that the hydrogel and its degradation by-products are noncytotoxic. The evaluation of the applicability of CH in the treatment of dermal burns in Wistar rats was performed by induction of full-thickness transcutaneous dermal wounds. Wound healing was monitored through macroscopic and histological analysis. From macroscopic analysis, the wound beds of the animals treated with CH were considerably smaller than those of the controls. Histological analysis revealed lack of a reactive or a granulomatous inflammatory reaction in skin lesions with CH and the absence of pathological abnormalities in the organs obtained by necropsy, which supported the local and systemic histocompatibility of the biomaterial. The present results suggest that this biomaterial may aid the re-establishment of skin architecture. PMID- 19903304 TI - Proteolytic activity in wound fluids and tissues derived from chronic venous leg ulcers. AB - Venous leg ulcers affect approximately 1% of the general population and 3.6% of those over the age of 65. The goal of the research described herein is to shorten the time to healing by developing wound care alternatives that are based on a comprehensive understanding of the venous ulcer wound environment. The proteolytic and inflammatory components in wound fluids and tissue biopsy samples were characterized in subjects with documented long-standing venous ulcers that had showed resistance to standard therapy. All wounds showed polymicrobial colonization with greater than 10(6) CFU/g. Myeloperoxidase, a measure of leukocyte infiltration, was also markedly elevated in these wounds. Zymography revealed the presence of both pro-matrix metalloproteinase (MMP)-2 and pro-MMP-9 in wound fluids and to a lesser extent in tissue biopsies. Using an immunocapture activity assay we reveal a sevenfold excess of MMP-9 in wound fluid as compared to tissue, with 73% in the activated form. In contrast, MMP-8 total protein levels were nearly equal in wound fluids and biopsies. Fibronectin, a critical component of the extracellular matrix, was shown to be degraded in both wound fluids and biopsy samples. Finally, the potential of a novel wound dressing to neutralize several constituents of this hostile wound environment is shown. PMID- 19903305 TI - Stimulated neovascularization, inflammation resolution and collagen maturation in healing rat cutaneous wounds by a heparan sulfate glycosaminoglycan mimetic, OTR4120. AB - Heparan sulfate glycosaminoglycans (HS-GAGs) are not only the structural elements of tissue architecture but also regulate the bioavailability and transduction pathways of heparan sulfate-bound polypeptides released by cells or the extracellular matrix. Heparan sulfate-bound polypeptides include inflammatory mediators, chemokines, angiogenic factors, morphogens, and growth-promoting factors that induce cell migration, proliferation, and differentiation in wound healing. OTR4120, a polymer engineered to mimic the properties of HS-GAGs, is used to replace the natural HS-GAGs that are degraded during wound repair, and enhance the tissue regeneration by preserving the cellular microenvironment and the endogenous signals needed for tissue regeneration. We previously demonstrated that OTR4120 treatment had a long-term effect on increasing breaking strength and vasodilation in healing rat full-thickness excisional wounds. The present study investigates the underlying mechanisms of the effects of OTR4120 treatment in improving the quality of cutaneous wound repair. We found that OTR4120 treatment stimulated inflammation resolution and increased neovascularization. OTR4120 treatment also promoted epidermal migration and proliferation during reepithelialization. Moreover, the granulation tissue formation and collagen maturation were improved in OTR4120-treated wounds. Three months after wounding, the effects of OTR4120 treatment on vascularization and inflammation resolution were normalized, except for an improved neodermis. We conclude that OTR4120 is a potential matrix therapeutic agent that ensures the quality of normal cutaneous wound repair and may restore impaired wound healing characterized by deficient angiogenesis and prolonged inflammation. PMID- 19903306 TI - Study on the debridement efficacy of formulated enzymatic wound debriding agents by in vitro assessment using artificial wound eschar and by an in vivo pig model. AB - An in vitro efficacy study using newly developed artificial wound eschar (AWE) substrate was conducted for assessing enzyme dose response. The AWE substrate is prepared by the enzymatic conversion of fibrinogen to fibrin in the presence of collagen, fibrin, and elastin to form an insoluble planar matrix. AWE substrate was placed on Franz Diffusion Cells for continuously monitoring the debridement progress. A parallel in vivo study was performed using pig thermal-burn wounds. Papain at concentrations of 200, 400, 800, and 1,600 U/mg was used as the model debriding enzyme for both studies. The data from the first 5 hours of the in vitro testing showed that debriding activity increased as the enzyme concentration increased. The histological results of the in vivo biopsy samples showed that enzyme doses above 800 and 1,600 U/mg successfully achieved debridement on day 8, while lower treatment groups still contained eschar tissue. Using the histological measurement results (wound depth score) a dose response that correlated to the in vitro assessment was found. Granulation tissue maturity and reepithelialization displayed correlation with the enzyme dose. Results indicate that AWE substrate can be used to predict debridement efficacy in vitro when correlation to the in vivo assessment is achieved. PMID- 19903308 TI - An examination of the supply and demand for clinical laboratory professionals in the United States. AB - Medical technologists (MTs) and technicians fill key roles in blood centers and hospital transfusion services. There is a concern that the number of new technologists and technicians entering the field is insufficient to keep up with demand created by the retirement of current personnel and continuing technological innovation. The American Society for Clinical Pathology reported in March 2009 that laboratories across the United States were struggling to fill MT and medical laboratory technician (MLT) positions. A survey of "positions available" advertisements from several months of the ABC Newsletter revealed that 35% were for MTs compared with 22% for physicians and 43% for all other positions. While not a scientific study, on the surface, this would indicate that blood centers are not immune from the struggle to fill positions within their laboratories. This paper will examine current trends in the United States as it relates to staffing for these positions and conclude by proposing actions that blood centers can take to help ensure an adequate supply of qualified MTs and MLTs in the future. PMID- 19903309 TI - Managing change. AB - BACKGROUND: In the 1980s, the Administrative Section Coordinating Committee (ASCC) published a series of administrative manuals to assist the American Association of Blood Banks (AABB) membership in developing processes for administrative activities conducted in a hospital or blood center. These manuals covered topics such as finance, inventory, information systems, and other areas that provide the infrastructure to support operations. In 2004, the ASCC adopted a plan to create a new set of documents addressing the AABB Quality System Essentials (QSEs) from an administrative viewpoint. In the past 30 years, blood banking has made significant strides in donor and patient safety with the addition of tests for infectious diseases, increased regulations, and the introduction of new technologies, as well as a focus on quality. The demands of managing and documenting all of these important advances and the frequency with which these changes have occurred have challenged our industry. STUDY DESIGN: While many articles and books are available for managing change in general, this article applies several aspects of the QSEs as a practical foundation for managing change in donor centers, hospital blood banks, and transfusion services. CONCLUSION: Utilizing this systematic guide to manage change can not only facilitate the change process but help to guard against unforeseen consequences. PMID- 19903310 TI - The benefits of supply chain integration using a third-party integrator. PMID- 19903311 TI - Case of the false document. PMID- 19903312 TI - Balancing agendas in donor testing. PMID- 19903313 TI - Kinematic classification of iliotibial band syndrome in runners. AB - Several inconsistent causative biomechanical factors are considered to be crucial in the occurrence of iliotibial band syndrome (ITBS). The focus of this study was on assessing differences in the kinematic characteristics between healthy runners [control group (CO)] and runners with ITBS in order to recommend treatment strategies to deal with this injury. Three-dimensional kinematics of barefoot running was used in the biomechanical setup. Both groups were matched with respect to gender, height and weight. After determining drop outs, the final population comprised 36 subjects (26 male and 10 female): 18 CO and 18 ITBS (13 male and five female, each). Kinematic evaluations indicate less hip adduction and frontal range of motion at the hip joint in runners with ITBS. Furthermore, maximum hip flexion velocity and maximum knee flexion velocity were lower in runners with ITBS. Lack of joint coordination, expressed as earlier hip flexion and a tendency toward earlier knee flexion, was found to be another discriminating variable in subjects with ITBS compared with CO subjects. We assume that an increase in range of motion at the hip joint, stretching of the hip abductors, as well as stretching the hamstrings, calf muscles and hip flexors will help treat ITBS. PMID- 19903314 TI - Self-admitted behavior and perceived use of performance-enhancing vs psychoactive drugs among competitive athletes. AB - The relationships between projected use, self-reported behavior and attitudes to performance-enhancing (PED) and recreational (RD) drugs were investigated among 82 competitive Hungarian athletes, with 14.6% admitting using PED and 31.7% using RD. Both the observed doping estimations (even those made by non-users) and self admitted use were considerably higher than the average rate of positive doping tests (2% of all tests). The notable overestimation by PED users (34.6% vs 16.9%) was in keeping with the false consensus effect. A prediction model with attitude and projection to the likelihood of PED use suggested at least a 70% chance of self-involvement of athletes, with responses at or above the median scores (Performance Enhancement Attitude Scale >= 60 and estimation >= 50%) on the two independent measures. Users overestimated the prevalence of doping in their sport (P=0.007) but not RD use, with the converse holding for RD users' views of doping (P=0.029). PED users also showed a significantly more lenient attitude toward doping (P<0.001). This domain-specific characteristic adds new information to the ongoing research effort in understanding drug-doping co-morbidity. The reasons for elevated in-group projection are discussed, along with the potential application of this phenomenon in doping epidemiology studies. PMID- 19903315 TI - Changes in the spatio-temporal organization of the trapezius muscle activity in response to eccentric contractions. AB - We hypothesized changes in the spatial organization of the trapezius muscle electromyographic (EMG) activity after a shoulder eccentric exercise. The rate of perceived exertion (RPE), the size of the soreness area, maximum force and, EMG from the upper, middle and lower trapezius were recorded. Root mean square (RMS), mean frequency (MNF) and normalized mutual information (a measure of functional connectivity between muscle sub-divisions) were computed during submaximal dynamic and static contractions performed before, immediately after and 24 h after exercise. Immediately after exercise, RPE, soreness area, RMS from the upper and middle trapezius and normalized mutual information among upper-middle sub-divisions increased while MNF decreased for the middle trapezius (P<0.05). After 24 h, the maximum force decreased. RMS from the upper trapezius and normalized mutual information among upper-middle trapezius sub-divisions were higher than before exercise. MNF values increased from immediately after to 24 h after for the upper and lower trapezius (P<0.05). The current results underlined changes in the spatio-temporal organization of the trapezius in response to shoulder eccentric exercise. The observed changes in EMG temporal and spectral contents and the enhanced sub-division coupling underlined the functional role of spatial variations of the EMG activity during muscle fatigue and in the presence of delayed-onset muscle soreness. PMID- 19903316 TI - Stress fracture in combination with avulsion from the tibia in a marathon runner: a case report. AB - We report an exceptional case of a tibial stress fracture combined with a large avulsion from the posterior facet of the tibia at the level where the tibialis posterior and soleus muscles insert. This type of injury has not been reported previously. A young healthy male runner experienced a sudden "snap" in the right lower leg 25 km into a marathon and had to leave the race due to increasing pain. The fracture was immediately apparent on plain radiographs. The injury was treated conservatively with 9 weeks in an ROM knee cast and no weight bearing on the affected leg, followed by full recovery. We propose that prolonged muscle tension at the site of tendinous attachment to the bone can cause the development of a stress fracture type of avulsion. PMID- 19903317 TI - Myostatin expression during human muscle hypertrophy and subsequent atrophy: increased myostatin with detraining. AB - Myostatin is a potent negative regulator of skeletal muscle mass, but its role in human skeletal muscle hypertrophy and atrophy is sparsely described. Muscle biopsies were obtained from young male subjects before and after 30 and 90 days of resistance training as well as after 3, 10, 30, 60 and 90 days of subsequent detraining. Myostatin mRNA increased significantly with detraining. We observed a 28 kDa myostatin immunoreactive protein, which, however, was also present in myostatin knock out mice skeletal muscle. As a novel finding we consistently detected a 10 kDa band, which may represent a mature myostatin monomer under reducing conditions or a novel, unknown myostatin form. Further, we observed a significant increase in this 10 kDa band after 3 days of detraining preceding the rapid type II fiber atrophy, in which almost half of the acquired fiber area was lost after only 10 days of detraining. Accordingly, an increase in the level of the 10 kDa protein is associated with rapid type II fiber atrophy, suggesting myostatin-mediated specific type II fiber atrophy, which in combination with our mRNA data support a role for myostatin in the negative regulation of adult human skeletal muscle mass. PMID- 19903318 TI - Neuromuscular fatigue differs following unilateral vs bilateral sustained submaximal contractions. AB - The purpose of the present study was to compare the mechanisms of fatigue induced by a unilateral vs a bilateral submaximal isometric knee extension. Ten physically active men completed two experimental sessions, randomly presented. They were asked to maintain an isometric knee extension force corresponding to 20% of the maximal voluntary contraction (MVC) until task failure with one leg (unilateral) vs two legs (bilateral). MVCs were performed before and after the sustained contraction. Transcutaneous electrical stimuli were used to examine central (voluntary activation) and peripheral (peak doublet force at rest) fatigue on the exercised leg. Time to task failure was significantly shorter (P<0.05) for the bilateral (245 +/- 76 s) compared with the unilateral task (295 +/- 85 s). Unilateral MVC force and maximal voluntary activation losses were significantly greater (P<0.05) after the unilateral task than after the bilateral task. Peak doublet force was significantly reduced (P<0.01) after the unilateral task, but not after the bilateral task. The present results demonstrated that time to task failure of a submaximal fatiguing contraction may depend on the number of limbs involved in the task. The greater time to task failure with one leg may have induced greater contractile alterations and a larger MVC loss following the unilateral task. PMID- 19903319 TI - Strength training improves 5-min all-out performance following 185 min of cycling. AB - To investigate the effects of heavy strength training on the mean power output in a 5-min all-out trial following 185 min of submaximal cycling at 44% of maximal aerobic power output in well-trained cyclists. Twenty well-trained cyclists were assigned to either usual endurance training combined with heavy strength training [E+S; n=11 (?=11)] or to usual endurance training only [E; n=9 (?=7, ?=2)]. The strength training performed by E+S consisted of four lower body exercises [3 * 4 10 repetition maximum (RM)], which were performed twice a week for 12 weeks. E+S increased 1 RM in half-squat (P<=0.001), while no change occurred in E. E+S led to greater reductions than E in oxygen consumption, heart rate, blood lactate concentration, and rate of perceived exertion (P<0.05) during the last hour of the prolonged cycling. Further, E+S increased the mean power output during the 5 min all-out trial (from 371 +/- 9 to 400 +/- 13 W, P<0.05), while no change occurred in E. In conclusion, adding strength training to usual endurance training improves leg strength and 5-min all-out performance following 185 min of cycling in well-trained cyclists. PMID- 19903320 TI - Detecting autologous blood transfusions: a comparison of three passport approaches and four blood markers. AB - Blood passport has been suggested as an indirect tool to detect various kinds of blood manipulations. Autologous blood transfusions are currently undetectable, and the objective of this study was to examine the sensitivities of different blood markers and blood passport approaches in order to determine the best approach to detect autologous blood transfusions. Twenty-nine subjects were transfused with either one (n=8) or three (n=21) bags of autologous blood. Hemoglobin concentration ([Hb]), percentage of reticulocytes (%ret) and hemoglobin mass (Hbmass) were measured 1 day before reinfusion and six times after reinfusion. The sensitivity and specificity of a novel marker, Hbmr (based on Hbmass and %ret), was evaluated together with [Hb], Hbmass and OFF-hr by different passport methods. Our novel Hbmr marker showed superior sensitivity in detecting the highest dosage of transfused blood, with OFF-hr showing equal or superior sensitivities at lower dosages. Hbmr and OFF-hr showed superior but equal sensitivities from 1 to 4 weeks after transfusion compared with [Hb] and Hbmass, with Hbmass being the only tenable prospect to detect acute transfusions. Because autologous blood transfusions can be an acute practice with blood withdrawal and reinfusion within a few days, Hbmass seems to be the only option for revealing this practice. PMID- 19903321 TI - Correlates of physical activity in Portuguese adolescents from 10 to 18 years. AB - This study examined the association between demographic [age, sex, socioeconomic status (SES)] and socio-cultural [father, mother, sibling physical activity (PA); peers and physical education teacher influences] correlates and low, moderate and high levels of PA among Portuguese adolescents aged 10-18 years. A total of 3352 males and females attending basic and secondary schools, their parents and siblings were sampled across four regions of Portugal. PA was assessed with a psychometrically validated questionnaire. Multinomial logistic regression was used. Age was positively related with moderate and high PA. Boys and adolescents of high SES were more likely to participate in moderate and high PA. Adolescents were more likely to participate in high PA when theirs mother and sibling(s) also participated. Peers had a positive influence on participation in moderate and high PA, while physical education teachers did not have an influence. The results indicated that demographic and socio-cultural correlates--in particular age, sex, SES, mother and sibling PA, and peer influence--were significantly associated with adolescent PA. These results also suggested that interventions should focus on girls and low SES adolescents who face higher risk of inactivity. PMID- 19903322 TI - Comparison of the QuantiFERON-TB Gold In-Tube test with the tuberculin skin test for detecting latent tuberculosis infection in hemodialysis patients. AB - BACKGROUND AND OBJECTIVES: Hemodialysis (HD) patients are at increased risk of reactivation of latent tuberculosis (TB) infection (LTBI). LTBI screening of this population is recommended. The QuantiFERON-TB Gold assay (QFT-G) may be more accurate than the tuberculin skin test (TST) in the detection of LTBI. We prospectively compared the results of QFT-G to TST in HD patients. METHODS: We examined 100 patients and performed TST and QFT-G tests. Data obtained from patients and medical records included medical history (past history of TB, Bacillus Calmette-Guerin [BCG] vaccination, history of contact with previous TB cases), radiography reports (chest x-ray with changes consistent with old TB), and basic laboratory findings. RESULTS: Forty-three of 100 patients (43%) had a positive QFT-G test result and 34 (34%) had a positive TST test result. Overall agreement between the QFT-G and the TST was 65% (concordance [k]=0.26, P=0.01). Discordant test results were seen in 13 TST-positive/QFT-G-negative patients and in 22 TST-negative/QFT-G-positive patients. Before BCG vaccination and radiographic reports (of old TB changes) were associated with discordant test results. On multivariate analysis, a positive QFT-G test was associated with contact with previous TB cases (P=0.026) and radiographic report (P=0.034), whereas a positive TST test also was associated with a history of BCG vaccination (P=0.015). CONCLUSIONS: QFT-G test results were more closely associated with TB risk factors than were positive TST results. Additionally, the QFT-G test was not affected by BCG vaccination. We concluded that QFT-G test is a more useful diagnostic method than TST for detecting LTBI in HD patients. PMID- 19903323 TI - Study of iron stores in regular plateletpheresis donors. AB - Plateletpheresis donors will lose up to 100 mL of blood at each donation, leading to concern that they may become iron deficient, particularly if donating at the maximum allowed frequency under National Blood Service policy of every 2 weeks. The serum ferritin levels of 508 regular plateletpheresis donors and 101 non donors were measured to indicate the level of their iron stores. About 33.9% (156/460) of platelet donors had depleted iron stores compared with 3.1% (3/97) non-donors. Results for male and post-menopausal female donors were similar with 36.2% (131/362) of males and 37.7% (20/53) of post-menopausal females showing iron depletion. There was clear correlation with donation frequency in males with 63.9% (46/72) of males donating at 2 weekly intervals found to be iron depleted. The percentage of iron depleted male subjects decreased as donation intervals increased. Correlation with lifetime donations of platelets was not demonstrated, although no donor who had given fewer than 14 blood and/or platelet donations was found to be iron depleted. In males there was a clear correlation between iron depletion and frequency of donation. There appeared to be no correlation with lifetime number of platelet donations. As a result of this study, we have advised that volunteers should not donate platelets more than 15 times per year, so that red cell loss is no more than the equivalent of three whole blood donations (1500 mL). PMID- 19903324 TI - The function of yeast epsin and Ede1 ubiquitin-binding domains during receptor internalization. AB - The formation of a primary endocytic vesicle is a dynamic process involving the transient organization of adaptor and scaffold proteins at the plasma membrane. Epsins and Eps15-like proteins are ubiquitin-binding proteins that act early in this process. The yeast epsins, Ent1 and Ent2, carry functional ubiquitin interacting motifs (UIMs), whereas the yeast Eps15-like protein, Ede1, has a C terminal ubiquitin-associated (UBA) domain. Analysis of mutants lacking early endocytic adaptors reveals that the ubiquitin-binding domains (UBDs) of Ent2 and Ede1 are likely to function primarily to mediate protein-protein interactions between components of the early endocytic machinery. Cells that lack epsin and Ede1 UBDs are able to internalize activated, ubiquitinated receptors. Furthermore, under conditions in which epsin UIMs are important for receptor internalization, receptors internalized via both ubiquitin-dependent and ubiquitin-independent signals require the UIMs, indicating that UIM function is not restricted to ubiquitinated receptors. Epsin UIMs share function with non-UBD protein-protein interaction motifs in Ent2 and Ede1, and the Ede1 UBA domain appears to negatively regulate interactions between endocytic proteins. Together, our results suggest that the ubiquitin-binding domains within the yeast epsin Ent2 and Ede1 are involved in the formation and regulation of the endocytic network. PMID- 19903325 TI - Immunocytokines: the long awaited therapeutic magic bullet in rheumatoid arthritis? AB - Modulatory cytokines such as IL-4 and IL-10 looked promising biologicals, but suffered from poor exposure at the inflamed joints when administered via the patient-friendly subcutaneous route. Immunocytokines have now been engineered with tissue targeting potential and are a possible solution to this problem, although challenges still exist. Local inflammatory processes cause destruction of extracellular matrix (ECM) components, leading to neo-eptitopes, and/or elicit the synthesis of new ECM components. This makes ECM elements interesting targets for antibody-mediated recognition and retention, to achieve higher levels of immunocytokines at the site of therapeutic interference. The study presented by Schwager and colleagues shows that targeted delivery of IL-10 is more efficacious in experimental arthritis. Clinical studies are warranted to show whether this strategy works for all rheumatoid arthritis patients or is better for subgroups with a defined ECM phenotype. In principle, the scFv-targeting system is plastic enough to allow for personalized strategies. PMID- 19903326 TI - An investigation of cognitive 'branching' processes in major depression. AB - BACKGROUND: Patients with depression demonstrate cognitive impairment on a wide range of cognitive tasks, particularly putative tasks of frontal lobe function. Recent models of frontal lobe function have argued that the frontal pole region is involved in cognitive branching, a process requiring holding in mind one goal while performing sub-goal processes. Evidence for this model comes from functional neuroimaging and frontal-pole lesion patients. We have utilised these new concepts to investigate the possibility that patients with depression are impaired at cognitive 'branching'. METHODS: 11 non-medicated patients with major depression were compared to 11 matched controls in a behavioural study on a task of cognitive 'branching'. In the version employed here, we recorded participant's performance as they learnt to perform the task. This involved participants completing a control condition, followed by a working memory condition, a dual task condition and finally the branching condition, which integrates processes in the working memory and dual-task conditions. We also measured participants on a number of other cognitive tasks as well as mood-state before and after the branching experiment. RESULTS: Patients took longer to learn the first condition, but performed comparably to controls after six runs of the task. Overall, reaction times decreased with repeated exposure on the task conditions in controls, with this effect attenuated in patients. Importantly, no differences were found between patients and controls on the branching condition. There was, however, a significant change in mood-state with patients increasing in positive affect and decreasing in negative affect after the experiment. CONCLUSION: We found no clear evidence of a fundamental impairment in anterior prefrontal 'branching processes' in patients with depression. Rather our data argue for a contextual learning impairment underlying cognitive dysfunction in this disorder. Our data suggest that MDD patients are able to perform high-level cognitive control tasks comparably to controls provided they are well trained. Future work should replicate these preliminary findings in a larger sample of MDD patients. PMID- 19903327 TI - A remarkable diversity of bone-eating worms (Osedax; Siboglinidae; Annelida). AB - BACKGROUND: Bone-eating Osedax worms have proved to be surprisingly diverse and widespread. Including the initial description of this genus in 2004, five species that live at depths between 25 and 3,000 m in the eastern and western Pacific and in the north Atlantic have been named to date. Here, we provide molecular and morphological evidence for 12 additional evolutionary lineages from Monterey Bay, California. To assess their phylogenetic relationships and possible status as new undescribed species, we examined DNA sequences from two mitochondrial (COI and 16S rRNA) and three nuclear genes (H3, 18S and 28S rRNA). RESULTS: Phylogenetic analyses identified 17 distinct evolutionary lineages. Levels of sequence divergence among the undescribed lineages were similar to those found among the named species. The 17 lineages clustered into five well-supported clades that also differed for a number of key morphological traits. Attempts to determine the evolutionary age of Osedax depended on prior assumptions about nucleotide substitution rates. According to one scenario involving a molecular clock calibrated for shallow marine invertebrates, Osedax split from its siboglinid relatives about 45 million years ago when archeocete cetaceans first appeared and then diversified during the late Oligocene and early Miocene when toothed and baleen whales appeared. Alternatively, the use of a slower clock calibrated for deep-sea annelids suggested that Osedax split from its siboglinid relatives during the Cretaceous and began to diversify during the Early Paleocene, at least 20 million years before the origin of large marine mammals. CONCLUSION: To help resolve uncertainties about the evolutionary age of Osedax, we suggest that the fossilized bones from Cretaceous marine reptiles and late Oligocene cetaceans be examined for possible trace fossils left by Osedax roots. Regardless of the outcome, the present molecular evidence for strong phylogenetic concordance across five separate genes suggests that the undescribed Osedax lineages comprise evolutionarily significant units that have been separate from one another for many millions of years. These data coupled with ongoing morphological analyses provide a solid foundation for their future descriptions as new species. PMID- 19903328 TI - Performance of optimized McRAPD in identification of 9 yeast species frequently isolated from patient samples: potential for automation. AB - BACKGROUND: Rapid, easy, economical and accurate species identification of yeasts isolated from clinical samples remains an important challenge for routine microbiological laboratories, because susceptibility to antifungal agents, probability to develop resistance and ability to cause disease vary in different species. To overcome the drawbacks of the currently available techniques we have recently proposed an innovative approach to yeast species identification based on RAPD genotyping and termed McRAPD (Melting curve of RAPD). Here we have evaluated its performance on a broader spectrum of clinically relevant yeast species and also examined the potential of automated and semi-automated interpretation of McRAPD data for yeast species identification. RESULTS: A simple fully automated algorithm based on normalized melting data identified 80% of the isolates correctly. When this algorithm was supplemented by semi-automated matching of decisive peaks in first derivative plots, 87% of the isolates were identified correctly. However, a computer-aided visual matching of derivative plots showed the best performance with average 98.3% of the accurately identified isolates, almost matching the 99.4% performance of traditional RAPD fingerprinting. CONCLUSION: Since McRAPD technique omits gel electrophoresis and can be performed in a rapid, economical and convenient way, we believe that it can find its place in routine identification of medically important yeasts in advanced diagnostic laboratories that are able to adopt this technique. It can also serve as a broad range high-throughput technique for epidemiological surveillance. PMID- 19903329 TI - Reduced levels of reactive oxygen species correlate with inhibition of apoptosis, rise in thioredoxin expression and increased bovine leukemia virus proviral loads. AB - BACKGROUND: Bovine Leukemia virus (BLV) is a deltaretrovirus that induces lymphoproliferation and leukemia in ruminants. In ex vivo cultures of B lymphocytes isolated from BLV-infected sheep show that spontaneous apoptosis is reduced. Here, we investigated the involvement of reactive oxygen species (ROS) in this process. RESULTS: We demonstrate that (i) the levels of ROS and a major product of oxidative stress (8-OHdG) are reduced, while the thioredoxin antioxidant protein is highly expressed in BLV-infected B lymphocytes, (ii) induction of ROS by valproate (VPA) is pro-apoptotic, (iii) inversely, the scavenging of ROS with N-acetylcysteine inhibits apoptosis, and finally (iv) the levels of ROS inversely correlate with the proviral loads. CONCLUSION: Together, these observations underline the importance of ROS in the mechanisms of inhibition of apoptosis linked to BLV infection. PMID- 19903330 TI - Comparative analysis of Panicum streak virus and Maize streak virus diversity, recombination patterns and phylogeography. AB - BACKGROUND: Panicum streak virus (PanSV; Family Geminiviridae; Genus Mastrevirus) is a close relative of Maize streak virus (MSV), the most serious viral threat to maize production in Africa. PanSV and MSV have the same leafhopper vector species, largely overlapping natural host ranges and similar geographical distributions across Africa and its associated Indian Ocean Islands. Unlike MSV, however, PanSV has no known economic relevance. RESULTS: Here we report on 16 new PanSV full genome sequences sampled throughout Africa and use these together with others in public databases to reveal that PanSV and MSV populations in general share very similar patterns of genetic exchange and geographically structured diversity. A potentially important difference between the species, however, is that the movement of MSV strains throughout Africa is apparently less constrained than that of PanSV strains. Interestingly the MSV-A strain which causes maize streak disease is apparently the most mobile of all the PanSV and MSV strains investigated. CONCLUSION: We therefore hypothesize that the generally increased mobility of MSV relative to other closely related species such as PanSV, may have been an important evolutionary step in the eventual emergence of MSV-A as a serious agricultural pathogen.The GenBank accession numbers for the sequences reported in this paper are GQ415386-GQ415401. PMID- 19903331 TI - Glutamate (mGluR-5) gene expression in brain regions of streptozotocin induced diabetic rats as a function of age: role in regulation of calcium release from the pancreatic islets in vitro. AB - Metabotrophic glutamate receptors (mGluRs) modulate cellular activities involved in the processes of differentiation and degeneration. In this study, we have analysed the expression pattern of group-I metabotropic glutamate receptor (mGlu 5) in cerebral cortex, corpus striatum, brainstem and hippocampus of streptozotocin induced and insulin treated diabetic rats (D+I) as a function of age. Also, the functional role of glutamate receptors in intra cellular calcium release from the pancreatic islets was studied in vitro. The gene expression studies showed that mGlu-5 mRNA in the cerebral cortex increased siginficantly in 7 weeks old diabetic rats whereas decreased expression was observed in brainstem, corpus striatum and hippocampus when compared to control. 90 weeks old diabetic rats showed decreased expression in cerebral cortex, corpus striatum and hippocampus whereas in brainstem the expression increased significantly compared to their respective controls. In 7 weeks old D+I group, mGlu-5 mRNA expression was significantly decreased in cerebral cortex and corpus striatum whereas the expression increased significantly in brainstem and hippocampus. 90 weeks old D+I group showed an increased expression in cerebral cortex, while it was decreased significantly in corpus striatum, brainstem and hippocampus compared to their respective controls. In vitro studies showed that glutamate at lower concentration (10(-7) M) stimulated calcium release from the pancreatic islets. Our results suggest that mGlu-5 receptors have differential expression in brain regions of diabetes and D+I groups as a function of age. This will have clinical significance in management of degeneration in brain function and memory enhancement through glutamate receptors. Also, the regulatory role of glutamate receptors in calcium release has immense therapeutic application in insulin secretion and function. PMID- 19903333 TI - Long-term follow-up of disability pensioners having musculoskeletal disorders. AB - BACKGROUND: Previously we have conducted a randomised controlled trial (RCT) to evaluate the effect of a brief cognitive behavioural program with a vocational approach aiming to return disability pensioners with back pain to work, as compared to no intervention. One year after the intervention, 10 participants (22%) who received the program and 5 (11%) in the control group reported to have entered a return to work process. The aims of this study were to evaluate long term effects of the intervention, and compare this effect to 2 reference populations not participating in the original trial. METHODS: Three groups of disability pensioners were investigated: 1) Disability pensioners having back pain (n = 89) previously participating in the RCT (randomized to either a brief cognitive behavioural intervention or to a control group), 2) 342 disability pensioners having back pain, but refusing to participate in the study and 3) 449 disability pensioners having other musculoskeletal disorders than back pain. Primary outcome was return to work, defined as a reduction in payment of disability pension. RESULTS: Only 2 of 89 (2.3%) participants from the RCT had reduced disability pension at 3-years follow-up, both from the control group. None of the participants that had been in a process of returning to work after 1 year had actually gained employment at 3-years follow-up. In the 2 groups not participating in the previous RCT, only 4 (1.2%) and 8 (1.6%) had returned to work after 3 years respectively. CONCLUSION: The number of pensioners who returned to work was negligible in all groups regardless of having participated in a cognitive behavioural intervention or not. PMID- 19903332 TI - Genomic transcriptional profiling identifies a candidate blood biomarker signature for the diagnosis of septicemic melioidosis. AB - BACKGROUND: Melioidosis is a severe infectious disease caused by Burkholderia pseudomallei, a Gram-negative bacillus classified by the National Institute of Allergy and Infectious Diseases (NIAID) as a category B priority agent. Septicemia is the most common presentation of the disease with a 40% mortality rate even with appropriate treatments. Better diagnostic tests are therefore needed to improve therapeutic efficacy and survival rates. RESULTS: We have used microarray technology to generate genome-wide transcriptional profiles (>48,000 transcripts) from the whole blood of patients with septicemic melioidosis (n = 32), patients with sepsis caused by other pathogens (n = 31), and uninfected controls (n = 29). Unsupervised analyses demonstrated the existence of a whole blood transcriptional signature distinguishing patients with sepsis from control subjects. The majority of changes observed were common to both septicemic melioidosis and sepsis caused by other infections, including genes related to inflammation, interferon-related genes, neutrophils, cytotoxic cells, and T cells. Finally, class prediction analysis identified a 37 transcript candidate diagnostic signature that distinguished melioidosis from sepsis caused by other organisms with 100% accuracy in a training set. This finding was confirmed in 2 independent validation sets, which gave high prediction accuracies of 78% and 80%, respectively. This signature was significantly enriched in genes coding for products involved in the MHC class II antigen processing and presentation pathway. CONCLUSIONS: Blood transcriptional patterns distinguish patients with septicemic melioidosis from patients with sepsis caused by other pathogens. Once confirmed in a large scale trial this diagnostic signature might constitute the basis of a differential diagnostic assay. PMID- 19903334 TI - The ATM and ATR inhibitors CGK733 and caffeine suppress cyclin D1 levels and inhibit cell proliferation. AB - The ataxia telangiectasia mutated (ATM) and the ATM- related (ATR) kinases play a central role in facilitating the resistance of cancer cells to genotoxic treatment regimens. The components of the ATM and ATR regulated signaling pathways thus provide attractive pharmacological targets, since their inhibition enhances cellular sensitivity to chemo- and radiotherapy. Caffeine as well as more specific inhibitors of ATM (KU55933) or ATM and ATR (CGK733) have recently been shown to induce cell death in drug-induced senescent tumor cells. Addition of these agents to cancer cells previously rendered senescent by exposure to genotoxins suppressed the ATM mediated p21 expression required for the survival of these cells. The precise molecular pharmacology of these agents however, is not well characterized. Herein, we report that caffeine, CGK733, and to a lesser extent KU55933, inhibit the proliferation of otherwise untreated human cancer and non-transformed mouse fibroblast cell lines. Exposure of human cancer cell lines to caffeine and CGK733 was associated with a rapid decline in cyclin D1 protein levels and a reduction in the levels of both phosphorylated and total retinoblastoma protein (RB). Our studies suggest that observations based on the effects of these compounds on cell proliferation and survival must be interpreted with caution. The differential effects of caffeine/CGK733 and KU55933 on cyclin D1 protein levels suggest that these agents will exhibit dissimilar molecular pharmacological profiles. PMID- 19903335 TI - Prednisolone Trial: Study protocol for a randomised controlled trial of prednisolone for women with idiopathic recurrent miscarriage and raised levels of uterine natural killer (uNK) cells in the endometrium. AB - BACKGROUND: Idiopathic recurrent miscarriage is defined as 3 consecutive pregnancy losses with no contributing features found on investigations. At present there are no treatments of proven efficacy for idiopathic recurrent miscarriage. Uterine natural killer (uNK) cells, the most predominant leucocyte in the endometrium are adjacent to foetal trophoblast cells and thought to be involved in implantation. The exact mechanisms of how uNK cells affect implantation are not clear but are probably through the regulation of angiogenesis. Multiple studies have shown an association between high density of uterine natural killer cells and recurrent miscarriage. We have shown that prednisolone reduces the number of uNK cells in the endometrium. The question remains as to whether reducing the number of uNK cells improves pregnancy outcome. METHODS: We propose a randomised, double-blind, placebo controlled trial of prednisolone with a pilot phase to assess feasibility of recruitment, integrity of trial procedures, and to generate data to base future power calculations. The primary aim is to investigate whether prednisolone therapy during the first trimester of pregnancy is able to improve live birth rates in patients with idiopathic recurrent miscarriage and raised uNK cells in the endometrium. Secondary outcomes include conception rate, karyotype of miscarriage, miscarriages (first and second trimester), stillbirths, pregnancy complications, gestational age at delivery, congenital abnormality and side effects of steroids. The trial has 2 stages: i) screening of non-pregnant women and ii) randomisation of the pregnant cohort. All patients who fit the inclusion criteria (<40 years old, > or =3 consecutive miscarriages with no cause found and no contraindications to prednisolone therapy) will be asked to consent to an endometrial biopsy in the mid-luteal phase to assess their levels of uNK cells. Women with high levels of uNK cells (> or =5%), will be randomised to either prednisolone or placebo when a pregnancy is confirmed. Follow-up includes 2 weekly ultrasound scans in the first trimester, an anomaly scan at 20 weeks gestation, growth scans at 28 and 34 weeks gestation and a postnatal follow-up at 6 weeks. TRIAL REGISTRATION: Current Controlled Trials ISRCTN28090716. PMID- 19903336 TI - Multivariate random effects meta-analysis of diagnostic tests with multiple thresholds. AB - BACKGROUND: Bivariate random effects meta-analysis of diagnostic tests is becoming a well established approach when studies present one two-by-two table or one pair of sensitivity and specificity. When studies present multiple thresholds for test positivity, usually meta-analysts reduce the data to a two-by-two table or take one threshold value at a time and apply the well developed meta-analytic approaches. However, this approach does not fully exploit the data. METHODS: In this paper we generalize the bivariate random effects approach to the situation where test results are presented with k thresholds for test positivity, resulting in a 2 by (k+1) table per study. The model can be fitted with standard likelihood procedures in statistical packages such as SAS (Proc NLMIXED). We follow a multivariate random effects approach; i.e., we assume that each study estimates a study specific ROC curve that can be viewed as randomly sampled from the population of all ROC curves of such studies. In contrast to the bivariate case, where nothing can be said about the shape of study specific ROC curves without additional untestable assumptions, the multivariate model can be used to describe study specific ROC curves. The models are easily extended with study level covariates. RESULTS: The method is illustrated using published meta-analysis data. The SAS NLMIXED syntax is given in the appendix. CONCLUSION: We conclude that the multivariate random effects meta-analysis approach is an appropriate and convenient framework to meta-analyse studies with multiple threshold without losing any information by dichotomizing the test results. PMID- 19903337 TI - FSL-1, a bacterial-derived toll-like receptor 2/6 agonist, enhances resistance to experimental HSV-2 infection. AB - BACKGROUND: Herpes simplex virus type 2 (HSV-2) is a leading cause of genital ulceration that can predispose individuals to an increased risk of acquiring other sexually transmitted infections. There are no approved HSV-2 vaccines and current suppressive therapies require daily compound administration that does not prevent all recurrences. A promising experimental strategy is the use of toll like receptor (TLR) agonists to induce an innate immune response that provides resistance to HSV-2 infection. Previous studies showed that anti-herpetic activity varied based on origin of the agonists and activation of different TLR indicating that activity likely occurs through elaboration of a specific innate immune response. To test the hypothesis, we evaluated the ability of a bacterial derived TLR2/6 agonist (FSL-1) to increase resistance to experimental genital HSV 2 infection. METHODS: Vaginal application of FSL-1 at selected doses and times was evaluated to identify potential increased resistance to genital HSV-2 infection in the mouse model. The FSL-1 induced cytokine profile was quantified using kinetically collected vaginal lavages. Additionally, cytokine elaboration and organ weights were evaluated after single or multiple FSL-1 doses to establish a preliminary safety profile. Human vaginal EC cultures were used to confirm the mouse model outcomes. RESULTS: The results showed that vaginally applied FSL-1 created an environment resistant to a 25-fold higher HSV-2 challenge dose. Mechanistically, vaginal FSL-1 application led to transient elaboration of cytokines linked to anti-herpetic innate immune responses. No gross local or peripheral immunotoxicity was observed even after multiple dosing. FSL-1 also created an anti-herpetic environment in cultures of human vaginal epithelial cells (EC). CONCLUSION: The results showed, for the first time, that the bacterial-derived TLR2/6 agonist FSL-1 induced significant resistance to HSV 2 infection when applied in mice or human vaginal EC cultures. Cytokine evaluation illustrated that anti-herpetic activity correlated with induction of a specific profile. The identified anti-herpetic profile provides an invaluable resource for the future design of novel compounds to reduce genital HSV-2 transmission and improves understanding of the complex innate immune response to potential pathogens elicited by the vaginal mucosa. PMID- 19903338 TI - Assessing the usefulness of a novel MRI-based breast density estimation algorithm in a cohort of women at high genetic risk of breast cancer: the UK MARIBS study. AB - INTRODUCTION: Mammographic breast density is one of the strongest known risk factors for breast cancer. We present a novel technique for estimating breast density based on 3D T1-weighted Magnetic Resonance Imaging (MRI) and evaluate its performance, including for breast cancer risk prediction, relative to two standard mammographic density-estimation methods. METHODS: The analyses were based on MRI (n = 655) and mammography (n = 607) images obtained in the course of the UK multicentre magnetic resonance imaging breast screening (MARIBS) study of asymptomatic women aged 31 to 49 years who were at high genetic risk of breast cancer. The MRI percent and absolute dense volumes were estimated using our novel algorithm (MRIBview) while mammographic percent and absolute dense area were estimated using the Cumulus thresholding algorithm and also using a 21-point Visual Assessment scale for one medio-lateral oblique image per woman. We assessed the relationships of the MRI and mammographic measures to one another, to standard anthropometric and hormonal factors, to BRCA1/2 genetic status, and to breast cancer risk (60 cases) using linear and Poisson regression. RESULTS: MRI percent dense volume is well correlated with mammographic percent dense area (R = 0.76) but overall gives estimates 8.1 percentage points lower (P < 0.0001). Both show strong associations with established anthropometric and hormonal factors. Mammographic percent dense area, and to a lesser extent MRI percent dense volume were lower in BRCA1 carriers (P = 0.001, P = 0.010 respectively) but there was no association with BRCA2 carrier status. The study was underpowered to detect expected associations between percent density and breast cancer, but women with absolute MRI dense volume in the upper half of the distribution had double the risk of those in the lower half (P = 0.009). CONCLUSIONS: The MRIBview estimates of volumetric breast density are highly correlated with mammographic dense area but are not equivalent measures; the MRI absolute dense volume shows potential as a predictor of breast cancer risk that merits further investigation. PMID- 19903339 TI - Gene expression profiling in sinonasal adenocarcinoma. AB - BACKGROUND: Sinonasal adenocarcinomas are uncommon tumors which develop in the ethmoid sinus after exposure to wood dust. Although the etiology of these tumors is well defined, very little is known about their molecular basis and no diagnostic tool exists for their early detection in high-risk workers. METHODS: To identify genes involved in this disease, we performed gene expression profiling using cancer-dedicated microarrays, on nine matched samples of sinonasal adenocarcinomas and non-tumor sinusal tissue. Microarray results were validated by quantitative RT-PCR and immunohistochemistry on two additional sets of tumors. RESULTS: Among the genes with significant differential expression we selected LGALS4, ACS5, CLU, SRI and CCT5 for further exploration. The overexpression of LGALS4, ACS5, SRI, CCT5 and the downregulation of CLU were confirmed by quantitative RT-PCR. Immunohistochemistry was performed for LGALS4 (Galectin 4), ACS5 (Acyl-CoA synthetase) and CLU (Clusterin) proteins: LGALS4 was highly up-regulated, particularly in the most differentiated tumors, while CLU was lost in all tumors. The expression of ACS5, was more heterogeneous and no correlation was observed with the tumor type. CONCLUSION: Within our microarray study in sinonasal adenocarcinoma we identified two proteins, LGALS4 and CLU, that were significantly differentially expressed in tumors compared to normal tissue. A further evaluation on a new set of tissues, including precancerous stages and low grade tumors, is necessary to evaluate the possibility of using them as diagnostic markers. PMID- 19903341 TI - Genome Alteration Print (GAP): a tool to visualize and mine complex cancer genomic profiles obtained by SNP arrays. AB - We describe a method for automatic detection of absolute segmental copy numbers and genotype status in complex cancer genome profiles measured with single nucleotide polymorphism (SNP) arrays. The method is based on pattern recognition of segmented and smoothed copy number and allelic imbalance profiles. Assignments were verified by DNA indexes of primary tumors and karyotypes of cell lines. The method performs well even for poor-quality data, low tumor content, and highly rearranged tumor genomes. PMID- 19903340 TI - PTEN inhibits BMI1 function independently of its phosphatase activity. AB - BACKGROUND: PTEN is the second most mutated tumor suppressor gene other than p53. It suppresses tumorigenesis by dephosphorylating phosphatidylinositol (3,4,5) triphosphate (PIP3) to phosphatidylinositol (4,5)-biphosphate (PIP2), thereby directly inhibiting phosphatidylinositol 3 kinase (PI3K)-mediated tumorigenic activities. Consistent with this model of action, cytosolic PTEN is recruited to the plasma membrane to dephosphorylate PIP3. While nuclear PTEN has been shown to suppress tumorigenesis by governing genome integrity, additional mechanisms may also contribute to nuclear PTEN-mediated tumor suppression. The nuclear protein BMI1 promotes stem cell self-renewal and tumorigenesis and PTEN inhibits these events, suggesting that PTEN may suppress BMI1 function. RESULTS: We investigated whether PTEN inhibits BMI1 function during prostate tumorigenesis. PTEN binds to BMI1 exclusively in the nucleus. This interaction does not require PTEN's phosphatase activity, as phosphatase-deficient PTEN mutants, PTEN/C124S (CS), PTEN/G129E (GE), and a C-terminal PTEN fragment (C-PTEN) excluding the catalytic domain, all associate with BMI1. Furthermore, the residues 186-286 of C-PTEN are sufficient for binding to BMI1. This interaction reduces BMI1's function. BMI1 enhances hTERT activity and reduces p16(INK4A) and p14(ARF) expression. These effects were attenuated by PTEN, PTEN(CS), PTEN(GE), and C-PTEN. Furthermore, knockdown of PTEN in DU145 cells increased hTERT promoter activity, which was reversed when BMI1 was concomitantly knocked-down, indicating that PTEN reduces hTERT promoter activity via inhibiting BMI1 function. Conversely, BMI1 reduces PTEN's ability to inhibit AKT activation, which can be attributed to its interaction with PTEN in the nucleus, making PTEN unavailable to dephosphorylate membrane-bound PIP3. Furthermore, BMI1 appears to co-localize with PTEN more frequently in clinical prostate tissue samples from patients diagnosed with PIN (prostatic intraepithelial neoplasia) and carcinoma compared to normal prostate epithelium. While PTEN co-localized with BMI1 in 2.4% of normal prostate epithelial cells, co-localization was observed in 37.6% and 18.5% of cells in PIN and carcinoma, respectively. Collectively, we demonstrate that PTEN inhibits BMI1 function via binding to BMI1 in a phosphatase independent manner. CONCLUSION: We demonstrate that nuclear PTEN reduces BMI1 function independently of its phosphatase activity. It was recently observed that nuclear PTEN also suppresses tumorigenesis. Our results, therefore, provide a plausible mechanism by which nuclear PTEN prevents tumorigenesis. PMID- 19903342 TI - Human spongiosa mesenchymal stem cells fail to generate cardiomyocytes in vitro. AB - BACKGROUND: Human mesenchymal stem cells (hMSCs) are broadly discussed as a promising cell population amongst others for regenerative therapy of ischemic heart disease and its consequences. Although cardiac-specific differentiation of hMSCs was reported in several in vitro studies, these results were sometimes controversial and not reproducible. RESULTS: In our study we have analyzed different published protocols of cardiac differentiation of hMSCs and their modifications, including the use of differentiation cocktails, different biomaterial scaffolds, co-culture techniques, and two- and three-dimensional cultures. We also studied whether 5'-azacytidin and trichostatin A treatments in combination with the techniques mentioned above can increase the cardiomyogenic potential of hMSCs. We found that hMSCs failed to generate functionally active cardiomyocytes in vitro, although part of the cells demonstrated increased levels of cardiac-specific gene expression when treated with differentiation factors, chemical substances, or co-cultured with native cardiomyocytes. CONCLUSION: The failure of hMSCs to form cardiomyocytes makes doubtful the possibility of their use for mechanical reparation of the heart muscle. PMID- 19903343 TI - Application of basic and composite thrombelastography parameters in monitoring of the antithrombotic effect of the low molecular weight heparin dalteparin: an in vivo study. AB - BACKGROUND: Low molecular weight heparin (LMWH) is in vast usage for treatment of thromboembolic diseases such as deep venous thrombosis and acute coronary syndromes. There are certain clinical situations where a quick point of care testing of the effect of LMWH would be useful. At this point there are no point of care devices available in the market for monitoring the effect of LMWH. Thrombelastography (TEG) evaluates the viscoelastic properties of blood during coagulation. The clinical application of TEG in monitoring LMWH treatment is not yet well defined. The purpose of this in vivo study was to systematically evaluate the most suitable TEG parameters for evaluation of the antithrombotic effect of LMWH. We furthermore evaluated for the first time the usefulness of the composite TEG parameter the Thrombodynamic Ratio (TDR) in monitoring LMWH treatment. METHODS: Healthy male volunteers (n = 7) were injected subcutaneously with the LMWH dalteparin 120 IU/kg. TEG parameters and antifactor Xa levels were measures at baseline, 2, 4, 5 and 24 hours after the injection. Correlation between TEG parameters and antiXa were calculated. The sensitivity and specificity of the TEG parameters for plasma levels of antiXa in the therapeutic range of 0.5 - 1.0 U/ml were calculated. RESULTS: All basic TEG parameters correlated significantly with antiXa levels. Among the basic parameters, the TEG reaction time R had the best correlation with antiXa levels with the most favorable combination of sensitivity and specificity for the therapeutic range of antiXa levels (r = 0.82, p < 0.0001, sensitivity 68%, specificity 100%). The composite TEG parameter TDR demonstrated the best correlation with antiXa levels, and an even more favorable combination of sensitivity and specificity compared to any of the basic parameters (r = - 0.87, p < 0.0001, sensitivity 95%, specificity 79%). CONCLUSION: The TEG reaction time R and TDR are the most suitable TEG parameters for evaluation of the antithrombotic effect of dalteparin with a highly significant correlation with antiXa levels in healthy male volunteers. Measures for uniform clinical use of these parameters are proposed. Larger clinical trials are needed to correlate R and TDR with clinical outcomes. PMID- 19903344 TI - Dietary intakes in infertile women a pilot study. AB - BACKGROUND: The reproductive axis is closely linked to nutritional status. The purpose of this study was to compare the nutritional status in two groups of young infertile women, without clinically overt eating disorders: hypothalamic amenorrhea (HA) and polycystic ovary syndrome (PCOS). METHODS: Eighteen young infertile women (10 HA, 8 PCOS) attending an outpatient gynecological endocrinology unit, underwent evaluation of anthropometry, body composition, dietary intakes by means of a food frequency questionnaire (FFQ) and a seven-day food diary (7DD), and psychological characteristics by means of EDI2 and SCL90 tests. RESULTS: HA women had lower BMI and body fat compared to PCOS women. Habitual intake derived from FFQs showed a similar macronutrient distribution between groups (about 16% protein, 33% fat, 52% carbohydrates). The psychometric profiles of the two groups did not differ significantly. The underreporting of dietary intakes (measured as habitual energy intake by FFQs/basal metabolic rate) was found to be negatively correlated with the interpersonal sensitivity SCL-90 subscale scores (r = -0.54, p = 0.02). CONCLUSION: Our study identified differences in body composition but not in dietary habits between HA and PCOS infertile women. We documented, for the first time, a relationship between the accuracy of dietary surveys and the psychological characteristics of subjects with anovulation. This finding suggests that it may be important to be aware of the psychological terrain when planning a dietary survey in infertile women. PMID- 19903345 TI - The Vital@Work Study. The systematic development of a lifestyle intervention to improve older workers' vitality and the design of a randomised controlled trial evaluating this intervention. AB - BACKGROUND: A major contributor of early exit from work is a decline in health with increasing age. As healthy lifestyle choices contribute to better health outcomes, an intervention aimed at an improved lifestyle is considered a potentially effective tool to keep older workers healthy and vital, and thereby to prolong labour participation. METHODS: Using the Intervention Mapping (IM) protocol, a lifestyle intervention was developed based on information obtained from 1) literature, 2) a short lifestyle questionnaire aimed at identifying the lifestyle behaviours among the target group, and 3) focusgroup (FG) interviews among 36 older workers (aged 45+ years) aimed at identifying: a) key determinants of lifestyle behaviour, b) a definition of vitality, and c) ideas about how vitality can be improved by lifestyle.The main lifestyle problems identified were: insufficient levels of physical activity and insufficient intake of fruit and vegetables. Using information from both literature and FG interviews, vitality consists of a mental and a physical component. The interviewees suggested to improve the mental component of vitality by means of relaxation exercises (e.g. yoga); physical vitality could be improved by aerobic endurance exercise and strength training.The lifestyle intervention (6 months) consists of three visits to a Personal Vitality Coach (PVC) combined with a Vitality Exercise Programme (VEP). The VEP consists of: 1) once a week a guided yoga group session aimed at relaxation exercises, 2) once a week a guided aerobic workout group session aimed at improving aerobic fitness and increasing muscle strength, and 3) older workers will be asked to perform once a week for at least 45 minutes vigorous physical activity without face-to-face instructions (e.g. fitness). Moreover, free fruit will be offered at the group sessions of the VEP. The lifestyle intervention will be evaluated in a RCT among older workers of two major academic hospitals in the Netherlands. At baseline, after 6 and 12 months, measurements (primary: lifestyle and vitality, and secondary: work-engagement and productivity) will take place. DISCUSSION: The lifestyle programme is developed specifically tailored to the needs of the older workers and which is aimed at improving their vitality. TRIAL REGISTRATION: NTR1240. PMID- 19903346 TI - Barking up the same tree: a comparison of ethnomedicine and canine ethnoveterinary medicine among the Aguaruna. AB - BACKGROUND: This work focuses on plant-based preparations that the Aguaruna Jivaro of Peru give to hunting dogs. Many plants are considered to improve dogs' sense of smell or stimulate them to hunt better, while others treat common illnesses that prevent dogs from hunting. This work places canine ethnoveterinary medicine within the larger context of Aguaruna ethnomedicine, by testing the following hypotheses: H1 -- Plants that the Aguaruna use to treat dogs will be the same plants that they use to treat people and H2 -- Plants that are used to treat both people and dogs will be used for the same illnesses in both cases. METHODS: Structured interviews with nine key informants were carried out in 2007, in Aguaruna communities in the Peruvian department of Amazonas. Informants provided freelists of plants given to dogs and explained the purpose, preparation and route of administration used. For each plant, informants also described any uses for treating people. Botanical voucher specimens were collected and additional informal observations were made, accompanying people on hunting trips. RESULTS: Out of 35 plant species given to dogs, 29 (83%) are also given to humans for some medicinal purpose, while five are used only for dogs. However, the same plant is used to treat the same illness in both humans and dogs in only 53% of the cases. Forty-three percent of plants used to treat a particular illness for both dogs and people are administered in the same manner for both. CONCLUSION: Results suggest that Aguaruna canine ethnoveterinary medicine is, at least partly, an independent cognitive domain. Some of the difference in plant use between dogs and people can be explained by the fact that certain diseases mentioned only apply to dogs. Although reports of canine ethnoveterinary medicine are very sparse in the literature, Aguaruna practices show some similarities with a few trends reported for other Amazonian societies, particularly, in the prevalence of the nasal route of administration, the use of plant-based psychoactives and in the importance of ants and wasps, in some form, for training dogs. PMID- 19903347 TI - A severe case of hemobilia and biliary fistula following an open urgent cholecystectomy. AB - BACKGROUND: Cholecystectomy has been the treatment of choice for symptomatic gallstones, but remains the greatest source of post-operative biliary injuries. Laparoscopic approach has been recently preferred because of short hospitalisation and low morbidity but has an higher incidence of biliary leakages and bile duct injuries than open one due to a technical error or misinterpretation of the anatomy. Even open cholecystectomy presents a small number of complications especially if it was performed in urgency. Hemobilia is one of the most common cause of upper gastrointestinal bleeding from the biliary ducts into the gastrointestinal tract due to trauma, advent of invasive procedures such as percutaneous liver biopsy, transhepatic cholangiography, and biliary drainage. METHODS: We report here a case of massive hemobilia in a 60 year-old man who underwent an urgent open cholecystectomy and a subsequent placement of a transhepatic biliary drainage. CONCLUSION: The management of these complications enclose endoscopic, percutaneous and surgical therapies. After a diagnosis of biliary fistula, it's most important to assess the adequacy of bile drainage to determine a controlled fistula and to avoid bile collection and peritonitis. Transarterial embolization is the first line of intervention to stop hemobilia while surgical intervention should be considered if embolization fails or is contraindicated. PMID- 19903348 TI - Dual stack black blood carotid artery CMR at 3T: application to wall thickness visualization. AB - BACKGROUND: The increasing understanding of atherosclerosis as an important risk factor for the development of acute ischemic events like ischemic stroke has stimulated increasing interest in non-invasive assessment of the structure, composition and burden of plaque depositions in the carotid artery wall. Vessel wall imaging by means of cardiovascular magnetic resonance (CMR) is conventionally done by 2D dual inversion recovery (DIR) techniques, which often fail in covering large volumes of interest as required in plaque burden assessment. Although the technique has been extended to 2D multislice imaging, its straight extension to 3D protocols is still limited by the prolonged acquisition times and incomplete blood suppression. A novel approach for rapid overview imaging of large sections of the carotid artery wall at isotropic spatial resolutions is presented, which omits excitation of the epiglottis. By the interleaved acquisition of two 3D stacks with the proposed motion sensitized segmented steady-state black-blood gradient echo technique (MSDS) the coverage of the carotid artery trees on both sides in reasonable scan times is enabled. RESULTS: 10 patients were investigated with the proposed technique and compared to conventional transversal DIR turbo spin and gradient echo approaches centered at the height of the carotid bifurcation. In all MSDS experiments sufficient black-blood contrast could be obtained over the entire covered volumes. The contrast to noise ratio between vessel and suppressed blood was improved by 73% applying the motion sensitizing technique. In all patients the suspicious areas of vessel wall thickening could be clearly identified and validated by the conventional local imaging approach. The average assessable vessel wall segment length was evaluated to be 18 cm. While in 50% of the cases motion artifacts could be appreciated in the conventional images, none were detected for the MSDS technique. CONCLUSION: The proposed technique enables the time efficient coverage of large areas of the carotid arteries without compromising wall-lumen CNR to get an overview about detrimental alterations of the vessel wall. Thickening of the vessel wall can be identified and the suspicious segments can be targeted for subsequent high-resolution CMR. The exclusion of the epiglottis may further facilitate reduction of swallowing induced motion artifacts. PMID- 19903349 TI - The role of community health workers in improving child health programmes in Mali. AB - BACKGROUND: Mortality of children under the age of five remains one of the most important public health challenges in developing countries. In rural settings, the promotion of household and community health practices through community health workers (CHWs) is among the key strategies to improve child health. The objective of this study was to assess the performance of CHWs in the promotion of basic child heath services in rural Mali. METHODS: A community-based cross sectional survey was undertaken using multi-stage cluster sampling of wards and villages. Data was collected through questionnaires among 401 child-caregivers and registers of 72 CHWs. RESULTS: Of 401 households suppose to receive a visit by a CHW, 219 (54.6%; confidence interval 95%; 49.6-59.5) had received at least one visit in the last three months before the survey. The mother is the most important caregiver (97%); high percentage being illiterate. Caregivers treat fever and diarrhoea with the correct regimen in 40% and 11% of cases respectively. Comparative analysis between households with and without CHW visits showed a positive influence of CHWs on family health practices: knowledge on the management of child fever (p = < 0.001), non-utilization of antibiotics in home treatment of diarrhoea (p = 0.003), presence of cloroquine in the household (p = 0.002), presence (p = 0.001) and use (p = < 0.001) of bed nets. A total of 27 (38%) CHWs had not received supervision at all, against 45 (63%) who have been followed regularly each month during the last six months. CONCLUSION: Continuous training, transport means, adequate supervision and motivation of CHWs through the introduction of financial incentives and remuneration are among key factors to improve the work of CHWs in rural communities. Poor performance of basic household health practices can be related to irregular supply of drugs and the need of appropriate follow-up by CHWs. PMID- 19903350 TI - Patterns of analgesic use, pain and self-efficacy: a cross-sectional study of patients attending a hospital rheumatology clinic. AB - BACKGROUND: Many people attending rheumatology clinics use analgesics and non steroidal anti-inflammatories for persistent musculoskeletal pain. Guidelines for pain management recommend regular and pre-emptive use of analgesics to reduce the impact of pain. Clinical experience indicates that analgesics are often not used in this way. Studies exploring use of analgesics in arthritis have historically measured adherence to such medication. Here we examine patterns of analgesic use and their relationships to pain, self-efficacy and demographic factors. METHODS: Consecutive patients were approached in a hospital rheumatology out-patient clinic. Pattern of analgesic use was assessed by response to statements such as 'I always take my tablets every day.' Pain and self-efficacy (SE) were measured using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and Arthritis Self-Efficacy Scale (ASES). Influence of factors on pain level and regularity of analgesic use were investigated using linear regression. Differences in pain between those agreeing and disagreeing with statements regarding analgesic use were assessed using t-tests. RESULTS: 218 patients (85% of attendees) completed the study. Six (2.8%) patients reported no current pain, 26 (12.3%) slight, 100 (47.4%) moderate, 62 (29.4%) severe and 17 (8.1%) extreme pain. In multiple linear regression self efficacy and regularity of analgesic use were significant (p < 0.01) with lower self efficacy and more regular use of analgesics associated with more pain.Low SE was associated with greater pain: 40 (41.7%) people with low SE reported severe pain versus 22 (18.3%) people with high SE, p < 0.001. Patients in greater pain were significantly more likely to take analgesics regularly; 13 (77%) of those in extreme pain reported always taking their analgesics every day, versus 9 (35%) in slight pain. Many patients, including 46% of those in severe pain, adjusted analgesic use to current pain level. In simple linear regression, pain was the only variable significantly associated with regularity of analgesic use: higher levels of pain corresponded to more regular analgesic use (p = 0.003). CONCLUSION: Our study confirms that there is a strong inverse relationship between self-efficacy and pain severity. Analgesics are often used irregularly by people with arthritis, including some reporting severe pain. PMID- 19903351 TI - MidExDB: a database of Drosophila CNS midline cell gene expression. AB - BACKGROUND: The Drosophila CNS midline cells are an excellent model system to study neuronal and glial development because of their diversity of cell types and the relative ease in identifying and studying the function of midline-expressed genes. In situ hybridization experiments generated a large dataset of midline gene expression patterns. To help synthesize these data and make them available to the scientific community, we developed a web-accessible database. DESCRIPTION: MidExDB (Drosophila CNS Midline Gene Expression Database) is comprised of images and data from our in situ hybridization experiments that examined midline gene expression. Multiple search tools are available to allow each type of data to be viewed and compared. Descriptions of each midline cell type and their development are included as background information. CONCLUSION: MidExDB integrates large scale gene expression data with the ability to identify individual cell types providing the foundation for detailed genetic, molecular, and biochemical studies of CNS midline cell neuronal and glial development and function. This information has general relevance for the study of nervous system development in other organisms, and also provides insight into transcriptional regulation. PMID- 19903352 TI - Altered serotonin physiology in human breast cancers favors paradoxical growth and cell survival. AB - INTRODUCTION: The breast microenvironment can either retard or accelerate the events associated with progression of latent cancers. However, the actions of local physiological mediators in the context of breast cancers are poorly understood. Serotonin (5-HT) is a critical local regulator of epithelial homeostasis in the breast and other organs. Herein, we report complex alterations in the intrinsic mammary gland serotonin system of human breast cancers. METHODS: Serotonin biosynthetic capacity was analyzed in human breast tumor tissue microarrays using immunohistochemistry for tryptophan hydroxylase 1 (TPH1). Serotonin receptors (5-HT1-7) were analyzed in human breast tumors using the Oncomine database. Serotonin receptor expression, signal transduction, and 5-HT effects on breast cancer cell phenotype were compared in non-transformed and transformed human breast cells. RESULTS: In the context of the normal mammary gland, 5-HT acts as a physiological regulator of lactation and involution, in part by favoring growth arrest and cell death. This tightly regulated 5-HT system is subverted in multiple ways in human breast cancers. Specifically, TPH1 expression undergoes a non-linear change during progression, with increased expression during malignant progression. Correspondingly, the tightly regulated pattern of 5-HT receptors becomes dysregulated in human breast cancer cells, resulting in both ectopic expression of some isoforms and suppression of others. The receptor expression change is accompanied by altered downstream signaling of 5-HT receptors in human breast cancer cells, resulting in resistance to 5-HT induced apoptosis, and stimulated proliferation. CONCLUSIONS: Our data constitutes the first report of direct involvement of 5-HT in human breast cancer. Increased 5-HT biosynthetic capacity accompanied by multiple changes in 5 HT receptor expression and signaling favor malignant progression of human breast cancer cells (for example, stimulated proliferation, inappropriate cell survival). This occurs through uncoupling of serotonin from the homeostatic regulatory mechanisms of the normal mammary epithelium. The findings open a new avenue for identification of diagnostic and prognostic markers, and valuable new therapeutic targets for managing breast cancer. PMID- 19903353 TI - Vitamin B6 deficient plants display increased sensitivity to high light and photo oxidative stress. AB - BACKGROUND: Vitamin B6 is a collective term for a group of six interconvertible compounds: pyridoxine, pyridoxal, pyridoxamine and their phosphorylated derivatives. Vitamin B6 plays essential roles as a cofactor in a range of biochemical reactions. In addition, vitamin B6 is able to quench reactive oxygen species in vitro, and exogenously applied vitamin B6 protects plant cells against cell death induced by singlet oxygen (1O2). These results raise the important question as to whether plants employ vitamin B6 as an antioxidant to protect themselves against reactive oxygen species. RESULTS: The pdx1.3 mutation affects the vitamin B6 biosynthesis enzyme, pyridoxal synthase (PDX1), and leads to a reduction of the vitamin B6 concentration in Arabidopsis thaliana leaves. Although leaves of the pdx1.3 Arabidopsis mutant contained less chlorophyll than wild-type leaves, we found that vitamin B6 deficiency did not significantly impact photosynthetic performance or shoot and root growth. Chlorophyll loss was associated with an increase in the chlorophyll a/b ratio and a selective decrease in the abundance of several PSII antenna proteins (Lhcb1/2, Lhcb6). These changes were strongly dependent on light intensity, with high light amplifying the difference between pdx1.3 and the wild type. When leaf discs were exposed to exogenous 1O2, lipid peroxidation in pdx1.3 was increased relative to the wild type; this effect was not observed with superoxide or hydrogen peroxide. When leaf discs or whole plants were exposed to excess light energy, 1O2-mediated lipid peroxidation was enhanced in leaves of the pdx1.3 mutant relative to the wild type. High light also caused an increased level of 1O2 in vitamin B6 deficient leaves. Combining the pdx1.3 mutation with mutations affecting the level of 'classical' quenchers of 1O2 (zeaxanthin, tocopherols) resulted in a highly photosensitive phenotype. CONCLUSION: This study demonstrates that vitamin B6 has a function in the in vivo antioxidant defense of plants. Thus, the antioxidant activity of vitamin B6 inferred from in vitro studies is confirmed in planta. Together with the finding that chloroplasts contain vitamin B6 compounds, the data show that vitamin B6 functions as a photoprotector that limits 1O2 accumulation in high light and prevents 1O2-mediated oxidative damage. PMID- 19903354 TI - Long-term prediction of fish growth under varying ambient temperature using a multiscale dynamic model. AB - BACKGROUND: Feed composition has a large impact on the growth of animals, particularly marine fish. We have developed a quantitative dynamic model that can predict the growth and body composition of marine fish for a given feed composition over a timespan of several months. The model takes into consideration the effects of environmental factors, particularly temperature, on growth, and it incorporates detailed kinetics describing the main metabolic processes (protein, lipid, and central metabolism) known to play major roles in growth and body composition. RESULTS: For validation, we compared our model's predictions with the results of several experimental studies. We showed that the model gives reliable predictions of growth, nutrient utilization (including amino acid retention), and body composition over a timespan of several months, longer than most of the previously developed predictive models. CONCLUSION: We demonstrate that, despite the difficulties involved, multiscale models in biology can yield reasonable and useful results. The model predictions are reliable over several timescales and in the presence of strong temperature fluctuations, which are crucial factors for modeling marine organism growth. The model provides important improvements over existing models. PMID- 19903355 TI - Impacts of feral horses on a desert environment. AB - BACKGROUND: Free-ranging horses (Equus caballus) in North America are considered to be feral animals since they are descendents of non-native domestic horses introduced to the continent. We conducted a study in a southern California desert to understand how feral horse movements and horse feces impacted this arid ecosystem. We evaluated five parameters susceptible to horse trampling: soil strength, vegetation cover, percent of nonnative vegetation, plant species diversity, and macroinvertebrate abundance. We also tested whether or not plant cover and species diversity were affected by the presence of horse feces. RESULTS: Horse trailing resulted in reduced vegetation cover, compacted soils, and in cases of intermediate intensity disturbance, increased plant species diversity. The presence of horse feces did not affect plant cover, but it did increase native plant diversity. CONCLUSION: Adverse impacts, such as soil compaction and increased erosion potential, were limited to established horse trails. In contrast, increased native plant diversity near trails and feces could be viewed as positive outcomes. Extensive trailing can result in a surprisingly large impact area: we estimate that < 30 horses used > 25 km2 of trails in our study area. PMID- 19903356 TI - Insulin-like growth factor binding protein-3 has dual effects on gastrointestinal stromal tumor cell viability and sensitivity to the anti-tumor effects of imatinib mesylate in vitro. AB - BACKGROUND: Imatinib mesylate has significantly improved survival and quality of life of patients with gastrointestinal stromal tumors (GISTs). However, the molecular mechanism through which imatinib exerts its anti-tumor effects is not clear. Previously, we found up-regulation of insulin-like growth factor binding protein-3 (IGFBP3) expression in imatinib-responsive GIST cells and tumor samples. Because IGFBP3 regulates cell proliferation and survival and mediates the anti-tumor effects of a number of anti-cancer agents through both IGF dependent and IGF-independent mechanisms, we hypothesized that IGFBP3 mediates GIST cell response to imatinib. To test this hypothesis, we manipulated IGFBP3 levels in two imatinib-responsive GIST cell lines and observed cell viability after drug treatment. RESULTS: In the GIST882 cell line, imatinib treatment induced endogenous IGFBP3 expression, and IGFBP3 down-modulation by neutralization or RNA interference resulted in partial resistance to imatinib. In contrast, IGFBP3 overexpression in GIST-T1, which had no detectable endogenous IGFBP3 expression after imatinib, had no effect on imatinib-induced loss of viability. Furthermore, both the loss of IGFBP3 in GIST882 cells and the overexpression of IGFBP3 in GIST-T1 cells was cytotoxic, demonstrating that IGFBP3 has opposing effects on GIST cell viability. CONCLUSION: This data demonstrates that IGFBP3 has dual, opposing roles in modulating GIST cell viability and response to imatinib in vitro. These preliminary findings suggest that there may be some clinical benefits to IGFBP3 therapy in GIST patients, but further studies are needed to better characterize the functions of IGFBP3 in GIST. PMID- 19903357 TI - Variation in genetic admixture and population structure among Latinos: the Los Angeles Latino eye study (LALES). AB - BACKGROUND: Population structure and admixture have strong confounding effects on genetic association studies. Discordant frequencies for age-related macular degeneration (AMD) risk alleles and for AMD incidence and prevalence rates are reported across different ethnic groups. We examined the genomic ancestry characterizing 538 Latinos drawn from the Los Angeles Latino Eye Study [LALES] as part of an ongoing AMD-association study. To help assess the degree of Native American ancestry inherited by Latino populations we sampled 25 Mayans and 5 Mexican Indians collected through Coriell's Institute. Levels of European, Asian, and African descent in Latinos were inferred through the USC Multiethnic Panel (USC MEP), formed from a sample from the Multiethnic Cohort (MEC) study, the Yoruba African samples from HapMap II, the Singapore Chinese Health Study, and a prospective cohort from Shanghai, China. A total of 233 ancestry informative markers were genotyped for 538 LALES Latinos, 30 Native Americans, and 355 USC MEP individuals (African Americans, Japanese, Chinese, European Americans, Latinos, and Native Hawaiians). Sensitivity of ancestry estimates to relative sample size was considered. RESULTS: We detected strong evidence for recent population admixture in LALES Latinos. Gradients of increasing Native American background and of correspondingly decreasing European ancestry were observed as a function of birth origin from North to South. The strongest excess of homozygosity, a reflection of recent population admixture, was observed in non-US born Latinos that recently populated the US. A set of 42 SNPs especially informative for distinguishing between Native Americans and Europeans were identified. CONCLUSION: These findings reflect the historic migration patterns of Native Americans and suggest that while the 'Latino' label is used to categorize the entire population, there exists a strong degree of heterogeneity within that population, and that it will be important to assess this heterogeneity within future association studies on Latino populations. Our study raises awareness of the diversity within "Latinos" and the necessity to assess appropriate risk and treatment management. PMID- 19903358 TI - A balanced t(5;17) (p15;q22-23) in chondroblastoma: frequency of the re arrangement and analysis of the candidate genes. AB - BACKGROUND: Chondroblastoma is a benign cartilaginous tumour of bone that predominantly affects the epiphysis of long bones in young males. No recurrent chromosomal re-arrangements have so far been observed. METHODS: We identified an index case with a balanced translocation by Combined Binary Ratio-Fluorescent in situ Hybridisation (COBRA-FISH) karyotyping followed by breakpoint FISH mapping and array-Comparative Genomic Hybridisation (aCGH). Candidate region re arrangement and candidate gene expression were subsequently investigated by interphase FISH and immunohistochemistry in another 14 cases. RESULTS: A balanced t(5;17)(p15;q22-23) was identified. In the index case, interphase FISH showed that the translocation was present only in mononucleated cells and was absent in the characteristic multinucleated giant cells. The t(5;17) translocation was not observed in the other cases studied. The breakpoint in 5p15 occurred close to the steroid reductase 5alpha1 (SRD5A1) gene. Expression of the protein was found in all cases tested. Similar expression was found for the sex steroid signalling related molecules oestrogen receptor alpha and aromatase, while androgen receptors were only found in isolated cells in a few cases. The breakpoint in 17q22-23 was upstream of the carbonic anhydrase x (CA10) gene region and possibly involved gene-regulatory elements, which was indicated by the lack of CA10 protein expression in the index case. All other cases showed variable levels of CA10 expression, with low expression in three cases. CONCLUSION: We report a novel t(5;17)(p15;q22-23) translocation in chondroblastoma without involvement of any of the two chromosomal regions in other cases studied. Our results indicate that the characteristic multinucleated giant cells in chondroblastoma do not have the same clonal origin as the mononuclear population, as they do not harbour the same translocation. We therefore hypothesise that they might be either reactive or originate from a distinct neoplastic clone, although the occurrence of two distinct clones is unlikely. Impairment of the CA10 gene might be pathogenetically relevant, as low expression was found in four cases. Diffuse expression of SRD5A1 and sex steroid signalling-related molecules confirms their role in neoplastic chondrogenesis. PMID- 19903359 TI - Enhanced detection and study of murine norovirus-1 using a more efficient microglial cell line. AB - BACKGROUND: Human Noroviruses are the predominant cause of non-bacterial gastroenteritis worldwide. To facilitate prevention and control, a norovirus isolated from mice can provide a model to understand human noroviruses. To establish optimal viral infectivity conditions for murine noroviruses, several cell lines of hematopoietic lineage, including murine BV-2, RAW 264.7, and TIB, as well as human CHME-5, were tested comparatively for their sensitivity to murine norovirus-1. RESULTS: Except for CHME-5, all three murine-derived cell lines were susceptible to MNV infection. Viral infection of these cells was confirmed by RT-PCR. Using both viral plaque and replication assays, BV-2 and RAW 264.7 cells were determined to have comparable sensitivities to MNV-1 infection. Comparisons of cell growth characteristics, general laboratory handling and potential in-field applications suggest the use of BV-2 to be more advantageous. CONCLUSION: Results obtained from these studies demonstrate that an immortalized microglial cell line can support MNV-1 replication and provides a more efficient method to detect and study murine noroviruses, facilitating future investigations using MNV-1 as a model to study, detect, and control Human Norovirus. PMID- 19903360 TI - B7-H4 gene polymorphisms are associated with sporadic breast cancer in a Chinese Han population. AB - BACKGROUND: B7-H4, a co-inhibitory molecule of the B7 family, can restrain T cell proliferation, cytokine secretion and the development of cytotoxicity. B7-H4 is expressed in tumor tissues at a higher level than in normal tissues, and has a potential effect to protect tumors from anti-tumor immune responses. This case control study was carried out to determine the potential influences of B7-H4 gene polymorphisms on the susceptibility and progression of breast cancer in Han women of Northeast China. METHODS: We genotyped three B7-H4 variants (rs10754339, rs10801935 and rs3738414) and tagged all common haplotypes (frequency greater than or equal to 1%) in a Chinese population consisting of 500 breast cancer cases and 504 control individuals matched for age. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) technique was used to determine the genotypes. RESULTS: Our data indicated that, compared with the common genotype and allele of each SNP, the rs10754339 AG genotype and G allele showed a significantly increased risk of breast cancer (OR = 1.455, 95% CI 1.119 1.892; OR = 1.325, 95% CI 1.073-1.637, respectively). The rs10801935 CC genotype, the rs3738414 AA genotype and the rs3738414 A allele were associated with a significantly decreased risk of breast cancer (OR = 0.328, 95% CI 0.145-0.739; OR = 0.412, 95% CI 0.203-0.835; OR = 0.698, 95% CI 0.564-0.864, respectively). Additionally, the rs10754339 GG genotype was significantly associated with lymph node metastasis and PR status, and the G allele and the AG genotype were respectively associated with lymph node metastasis and ER status. In haplotype analysis, we observed that compared with the AAG haplotype, the AAA haplotype showed a significantly decreased risk of breast cancer (OR = 0.689, 95% CI 0.539 0.881), but the GAG haplotype was associated with a significantly increased risk of breast cancer (OR = 1.511, 95% CI 1.125-2.031). And the AAA and the GCG haplotypes also respectively have significant influences on tumor size and ER status. CONCLUSION: These results suggest that B7-H4 gene polymorphism may contribute to the sporadic breast cancer risk and prognosis in Chinese Han women. PMID- 19903361 TI - Illumina WG-6 BeadChip strips should be normalized separately. AB - BACKGROUND: Illumina Sentrix-6 Whole-Genome Expression BeadChips are relatively new microarray platforms which have been used in many microarray studies in the past few years. These Chips have a unique design in which each Chip contains six microarrays and each microarray consists of two separate physical strips, posing special challenges for precise between-array normalization of expression values. RESULTS: None of the normalization strategies proposed so far for this microarray platform allow for the possibility of systematic variation between the two strips comprising each array. That this variation can be substantial is illustrated by a data example. We demonstrate that normalizing at the strip-level rather than at the array-level can effectively remove this between-strip variation, improve the precision of gene expression measurements and discover more differentially expressed genes. The gain is substantial, yielding a 20% increase in statistical information and doubling the number of genes detected at a 5% false discovery rate. Functional analysis reveals that the extra genes found tend to have interesting biological meanings, dramatically strengthening the biological conclusions from the experiment. Strip-level normalization still outperforms array-level normalization when non-expressed probes are filtered out. CONCLUSION: Plots are proposed which demonstrate how the need for strip-level normalization relates to inconsistent intensity range variation between the strips. Strip-level normalization is recommended for the preprocessing of Illumina Sentrix-6 BeadChips whenever the intensity range is seen to be inconsistent between the strips. R code is provided to implement the recommended plots and normalization algorithms. PMID- 19903362 TI - Two-dimensional nanosecond electric field mapping based on cell electropermeabilization. AB - Nanosecond, megavolt-per-meter electric pulses cause permeabilization of cells to small molecules, programmed cell death (apoptosis) in tumor cells, and are under evaluation as a treatment for skin cancer. We use nanoelectroporation and fluorescence imaging to construct two-dimensional maps of the electric field associated with delivery of 15 ns, 10 kV pulses to monolayers of the human prostate cancer cell line PC3 from three different electrode configurations: single-needle, five-needle, and flat-cut coaxial cable. Influx of the normally impermeant fluorescent dye YO-PRO-1 serves as a sensitive indicator of membrane permeabilization. The level of fluorescence emission after pulse exposure is proportional to the applied electric field strength. Spatial electric field distributions were compared in a plane normal to the center axis and 15-20 mum from the tip of the center electrode. Measurement results agree well with models for the three electrode arrangements evaluated in this study. This live-cell method for measuring a nanosecond pulsed electric field distribution provides an operationally meaningful calibration of electrode designs for biological applications and permits visualization of the relative sensitivities of different cell types to nanoelectropulse stimulation. PACS Codes: 87.85.M- PMID- 19903363 TI - Neuroticism as a common dimension in the internalizing disorders. AB - BACKGROUND: Several theories have posited a common internalizing factor to help account for the relationship between mood and anxiety disorders. These disorders are often co-morbid and strongly covary. Other theories and data suggest that personality traits may account, at least in part, for co-morbidity between depression and anxiety. The present study examined the relationship between neuroticism and an internalizing dimension common to mood and anxiety disorders. METHOD: A sample of ethnically diverse adolescents (n=621) completed self-report and peer-report measures of neuroticism. Participants also completed the Structured Clinical Interview for DSM-IV (SCID). RESULTS: Structural equation modeling showed that a single internalizing factor was common to lifetime diagnosis of mood and anxiety disorders, and this internalizing factor was strongly correlated with neuroticism. Neuroticism had a stronger correlation with an internalizing factor (r=0.98) than with a substance use factor (r=0.29). Therefore, neuroticism showed both convergent and discriminant validity. CONCLUSIONS: These results provide further evidence that neuroticism is a necessary factor in structural theories of mood and anxiety disorders. In this study, the correlation between internalizing psychopathology and neuroticism approached 1.0, suggesting that neuroticism may be the core of internalizing psychopathology. Future studies are needed to examine this possibility in other populations, and to replicate our findings. PMID- 19903364 TI - Affective symptoms as predictors of Alzheimer's disease in subjects with mild cognitive impairment: a 10-year follow-up study. AB - BACKGROUND: Affective symptoms are common in subjects with mild cognitive impairment (MCI), but there is disagreement whether these symptoms are predictive for Alzheimer's disease (AD). We investigated the predictive accuracy of affective symptoms for AD during a follow-up study in subjects with MCI, and whether the predictive accuracy was modified by age, the presence of amnestic MCI or the length of follow-up. METHOD: Newly referred subjects (n=263) with MCI older than 55 years were selected from a memory clinic and followed up after 2, 5 and 10 years. Predictors investigated were: symptoms of depression, anxiety, apathy and sleeping problems. RESULTS: Affective symptoms were present in 50-70% of the subjects. The average follow-up period was 5.4 years and 79 subjects (29%) developed AD. Sleeping problems were associated with a decreased risk for AD [odds ratio (OR) 0.35, p<0.001]. Symptoms of depression (OR 0.61, p=0.059) and anxiety (OR 0.58, p=0.051) showed a trend in the same direction. The OR of apathy for AD was 0.67 (p=0.14). Depression was associated with a decreased risk for AD only in subjects without amnestic MCI, but not in subjects with amnestic MCI. Moreover, anxiety was related to the risk for AD differently between subjects diagnosed with AD at the 5-year follow-up (OR 0.23) and subjects diagnosed with AD at the 10-year follow-up (OR 1.7). CONCLUSIONS: Affective symptoms are associated with a decreased risk for AD. The risk may be dependent on MCI subtype or length of follow-up, but it does not depend on age. PMID- 19903365 TI - Research letter: Psychotherapy increases brain serotonin 5-HT1A receptors in patients with major depressive disorder. PMID- 19903366 TI - Job insecurity, socio-economic circumstances and depression. AB - BACKGROUND: Economic recessions are characterized by job insecurity and rising unemployment. The relationship between job insecurity and poor mental health is known. However, we do not know how this relationship is affected by individual socio-economic circumstances. METHOD: A random probability sample comprising 3581 respondents (1746 men and 1835 women) were selected from the third national survey of psychiatric morbidity in Great Britain. Fieldwork was carried out throughout 2007. Depression was assessed using the revised Clinical Interview Schedule and ICD-10 research diagnostic criteria administered by well-trained lay interviewers. RESULTS: One-fifth of all working men and women aged 16-64 years felt that their job security was poor. From a multivariate analysis of several job stressors, there was an increased likelihood of depression among those agreeing that their job security was poor [odds ratio (OR) 1.58, 95% confidence intervals (CI) 1.22-2.06, p<0.001]. After controlling for age and sex, job insecurity (OR 1.86, 95% CI 1.47-2.35, p<0.001) and being in debt (OR 2.17, 95% CI 1.58-2.98, p<0.001) were independently associated with depression. CONCLUSIONS: Job insecurity has a strong association with feelings of depression even after controlling for biographic characteristics (age and sex), economic factors (personal debt) and work characteristics (type of work and level of responsibility). Despite the organizational changes needed to cope with a recession, employers should also take note of the additional distress experienced by workers at a time of great uncertainty, particularly those in less skilled jobs and in financial straits. PMID- 19903367 TI - Antimalarial compounds isolated from plants used in traditional medicine. AB - OBJECTIVES: This review covers the compounds with antiplasmodial activity isolated from plants published from 2005 to the end of 2008, organized according to their phytochemical classes. Details are given for substances with IC50 values < or = 11 microm. KEY FINDINGS: Malaria is a major parasitic disease in many tropical and subtropical regions and is responsible for more than 1 million deaths each year in Africa. The rapid spread of resistance encourages the search for new active compounds. Nature and particularly plants used in traditional medicine are a potential source of new antimalarial drugs as they contain molecules with a great variety of structures and pharmacological activities. SUMMARY: A large number of antimalarial compounds with a wide variety of structures have been isolated from plants and can play a role in the development of new antimalarial drugs. Ethnopharmacological approaches appear to be a promising way to find plant metabolites that could be used as templates for designing new derivatives with improved properties. PMID- 19903368 TI - Non-phenolic radical-trapping antioxidants. AB - OBJECTIVES: The aim of this review article is to introduce the reader to the mechanisms, rates and thermodynamic aspects of the processes involving the most biologically relevant non-phenolic radical-trapping antioxidants. KEY FINDINGS: Antioxidant defences in living organisms rely on a complex interplay between small molecules and enzymes, which cooperate in regulating the concentrations of potentially harmful oxidizing species within physiological limits. The noxious effects of an uncontrolled production of oxygen- and nitrogen-centered radicals are amplified by chain reactions (autoxidations), sustained mainly by peroxyl radicals (ROO(*)), that oxidize and alter essential biomolecules such as lipids, lipoproteins, proteins and nucleic acids. SUMMARY: Non-phenolic antioxidants represent an important and abundant class of radical scavengers in living organisms. These compounds react with peroxyl radicals through various mechanisms: (i) formal H-atom donation from weak X-H bonds (X = O, N, S), as in the case of ascorbic acid (vitamin C), uric acid, bilirubin and thiols; (ii) addition reactions to polyunsaturated systems with formation of C-radicals poorly reactive towards O(2), for example beta-carotene and all carotenoids in general; (iii) co-oxidation processes characterized by fast cross-termination reactions, for example gamma-terpinene; and (iv) catalytic quenching of superoxide (O(2)(* )) with a superoxide dismutase-like mechanism, for example di-alkyl nitroxides and FeCl(3). Kinetic data necessary to evaluate and rationalize the effects of these processes are reported. The mechanisms underlying the pro-oxidant effects of ascorbate and other reducing agents are also discussed. PMID- 19903369 TI - Modelling of diclofenac sodium diffusion from swellable and water-soluble polyethylene oxide matrices. AB - OBJECTIVES: The main objective of this study was to develop a mathematical model for the characterization of diclofenac sodium diffusion from polyethylene oxide (PEO) matrices. A model was developed on the basis of the diffusion theory accounting for the characteristics of the polymer: swelling with subsequent dissolution in water. The concentration-dependent diffusion of drug and water was taken into account. Experimental data were analysed using a computer software program as an aid for solving partial differential equations. METHODS: Six formulations of matrix tablets with different drug-excipient ratios were prepared using low-molecular-weight PEO as a matrix-forming material. For obtaining drug release data, dissolution studies were performed and water uptake by pure PEO matrices was studied as well. KEY FINDINGS: A good agreement of the developed model with experimental results was demonstrated. Some anomalies in drug diffusion were observed and their origin was questioned. Changes in the parameters characterizing the process of diffusion are attributed to glassy rubbery polymer transitions. Additional interpretation of this phenomenon on the basis of percolation theory is also provided. CONCLUSIONS: The obtained model has the ability to predict the required characteristics of matrices for desired drug release. The composition of batches with undesirable release properties can be predetermined and avoided in manufacturing. PMID- 19903370 TI - Effective vaginal DNA delivery with high transfection efficiency is a good system for induction of higher local vaginal immune responses. AB - OBJECTIVES: To investigate the local vaginal and systemic immune responses of effective vaginal DNA delivery with high transfection efficiency, we determined the effects on Th1-dependent cytokine (interferon-gamma) production in spleen and inguinal lymph node cells and antibody responses of vaginal pDNA immunization with a cell-penetrating peptide, and compared our vaginal immunization with intradermal and intranasal immunizations. METHODS: Mice were immunized by vaginal, nasal or dermal administration of pCMV-OVA with or without peptide carriers, and serum, vaginal fluids, spleen and inguinal cells were harvested. The serum immunoglobulin (Ig)G(2a) and vaginal IgA antibody responses were determined by sandwich enzyme-linked immunosorbent assay (ELISA). The interferon gamma production from spleen cells or inguinal lymph node cells was determined by an ELISA kit. KEY FINDINGS: The direct vaginal immunization strongly induced IgA in the vaginal fluids and interferon-gamma production in the local lymph node draining from the vagina. In addition, co-vaccination with the peptide carriers elevated these immune responses compared with vaccination with pCMV-OVA alone. Vaginal immunization with high transfection efficiency promoted vaginal IgA production to a significantly greater extent than intradermal or nasal immunization. CONCLUSIONS: These results suggested that direct vaginal DNA vaccines under high transfection conditions induced higher local vaginal antibody than that by intranasal or intradermal administration, and peptide carriers effectively elevated mucosal immune responses. Therefore, this vaginal DNA vaccination method may be expected to be useful in the prevention and treatment methods for vaginal infectious diseases such as HIV infection. PMID- 19903371 TI - Evaluation of molecular pharmaceutical and in-vivo properties of spray-dried isolated andrographolide-PVP. AB - OBJECTIVES: Andrographolide, a natural lipophilic molecule, has a wide range of pharmacological actions. However, due to low aqueous solubility, it has low oral bioavailability. The purpose of the study was to increase the solubility and dissolution rate of isolated andrographolide by formulating its solid dispersion. METHOD: Solid dispersions were obtained by a spray-drying technique using different ratios of drug to polyvinylpyrrolidine (PVP K-30). Solid dispersions in compression with isolated drug and corresponding physical mixtures were characterized for various molecular pharmaceutical properties and subjected to stability study for up to 3 months. KEY FINDINGS: A five-fold increase in saturation solubility of andrographolide with higher values of Q(5 min) (cumulative percentage release in 5 min) and lower values of t(75%) (time required for 75% w/w drug release) for solid dispersion was observed in different dissolution mediums. This was attributed to the formation of amorphous nature and intermolecular hydrogen bonding between drug and PVP K-30. The stability study showed there to be no significant change in molecular pharmaceutical properties and dissolution profile over the period of 3 months. Moreover, the in-vivo study in Wistar albino rats also justified improvement in the therapeutic efficacy of andrographolide after solid dispersion. CONCLUSIONS: This study demonstrates the utility of solid dispersion to improve primary and secondary pharmaceutical properties of andrographolide using PVP K-30 as a carrier. PMID- 19903372 TI - Quercetin and naringenin transport across human intestinal Caco-2 cells. AB - OBJECTIVES: Flavonoids are phenolic compounds found in most edible fruits and vegetables. Previous studies have demonstrated their biological and beneficial effects on human health. However, their bioavailability and, in particular, their intestinal absorption mechanism have not yet been clearly identified. The aim of our work was to quantify and to characterize in vitro the nature of the transport of two flavonoids distinguished by their physicochemical and pharmacological properties: quercetin, a flavan-3-ol, and naringenin, a flavanone. METHODS: Differentiated and polarized Caco-2 human intestinal epithelial cell lines were used for this purpose. KEY FINDINGS: In our experimental conditions, quercetin and naringenin were poorly absorbed by Caco-2 cells. Quercetin was absorbed by passive diffusion and a pH-dependent mechanism mediated by the organic anion transporting protein B (OATP-B). It was not a multidrug resistance associated protein (MRP)1 substrate, but was substrate of the MRP2 efflux transporter and not P-glycoprotein (P-gp). Intestinal permeability from the apical to the basolateral side was higher for naringenin than for quercetin, which was partly explained by naringenin's physicochemical characteristics. Naringenin, partially absorbed by passive diffusion, was also an ATP-dependent transport substrate mediated by MRP1, but was not an OATP-B substrate. However, naringenin was secreted via active P-gp and MRP2 efflux transporters. CONCLUSIONS: The contribution of ATP-dependent efflux transporters (MRP2 and P-gp) to the permeability of these compounds in the apical side could explain their low bioavailability. In conclusion, knowledge of the absorption mechanism of these two flavonoids was used to determine the intake level that has a beneficial effect on human health and their putative role in food-drug interactions. PMID- 19903373 TI - Preparation and pharmacokinetics of docetaxel based on nanostructured lipid carriers. AB - OBJECTIVES: This study describes the preparation and pharmacokinetics of docetaxel based on freeze-dried nanostructured lipid carriers (NLCs). METHODS: The docetaxel-incorporated NLCs were developed using hot high-pressure homogenisation, and lyophilised to obtain freeze-dried docetaxel NLCs. The influences of different concentrations of lipid matrices, ratio of drug to lipid, and different cryoprotectants on the characteristics of the NLCs were investigated. KEY FINDINGS: Freeze-dried docetaxel NLCs were spherical, with 5% (w/w) docetaxel loading efficiency and were stable for at least 6 months at 25 masculineC. X-ray powder diffraction and differential scanning calorimetry analysis suggested that docetaxel was distributed in a molecular or amorphous status. In-vitro release studies showed sustained drug release, with the cumulated release rate of 13% within 24 h without burst release. The freeze-dried docetaxel NLCs also showed sustained-release properties after intravenous injection into rats. The area under the plasma-concentration time curve and mean residence time were increased 4.90 and 2.82 times compared with docetaxel solution. The concentration of docetaxel in the lungs was significantly higher in rats treated with the NLCs than in those given docetaxel solution. CONCLUSIONS: Docetaxel NLCs have an organ-targeting effect and prolonged mean retention time and have potential for the treatment of lung cancer. PMID- 19903374 TI - Involvement of adipokines in rimonabant-mediated insulin sensitivity in ob/ob mice. AB - OBJECTIVES: It has been recently reported that blockade of type 1 cannabinoid (CB1) receptors by specific antagonists or genetic manipulation alleviates dyslipidaemia, hyperglycaemia and insulin resistance in animal models of obesity and type 2 diabetes. However, the precise role of adipokines in the insulin sensitising effects of the CB1 antagonist rimonabant is not clear. METHODS: ob/ob mice were treated with different doses of rimonabant and then subjected to an oral glucose tolerance test. The expression of different adipokines in white adipose tissue was analysed by quantitative real-time PCR. KEY FINDINGS: Rimonabant (30 mg/kg) significantly inhibited body weight and fat pad weight gain (P < 0.05) and improved glucose tolerance. Gene expression analysis indicated that tumour necrosis factor-alpha, visfatin and retinol binding protein-4 were downregulated in the adipose tissue of ob/ob mice treated with rimonabant compared with controls, whereas adiponectin was significantly upregulated. CONCLUSIONS: Rimonabant-mediated alteration of adipokines in white adipose tissues may play a role in improving insulin sensitivity in obese animals. PMID- 19903375 TI - Montmorillonite adsorbs uric acid and increases the excretion of uric acid from the intestinal tract in mice. AB - OBJECTIVES: The aim was to evaluate the adsorbing effect of montmorillonite on uric acid, promoting diffusion of uric acid from blood to intestine, preventing absorption of uric acid in intestine and reducing uric acid level in serum. METHODS: The adsorbing effect of montmorillonite on uric acid was observed in vitro. The intestine and blood vessel of rats were circularly perfused with intestinal perfusate and vascular perfusate, respectively. A model of hyperuricaemia in mice was prepared by intraperitoneal injection of hypoxanthine and potassium oteracil. The concentration of uric acid was determined by the method of urate oxidase and peroxide enzyme. KEY FINDINGS: The results showed that different concentrations of montmorillonite could adsorb uric acid in a concentration-dependent manner. The adsorbing effect was fast. The adsorptive rate was high in acid solution and was low in alkaline solution. When blood vessels were circularly perfused by vascular perfusate containing uric acid, the concentration of uric acid in vascular perfusate was decreased and the concentration of uric acid in intestinal perfusate was increased, suggesting that uric acid in blood vessels diffused into the intestine. When the intestine was perfused with intestinal perfusate containing uric acid, the uric acid concentration in vascular perfusate was increased, but the uric acid concentration of intestinal perfusate was decreased, suggesting that uric acid was absorbed in the intestine. The uric acid concentrations of intestinal perfusate and vascular perfusate in montmorillonite 0.5 and 1.0 g/kg groups were lower than the control group. Concentrations of uric acid in serum and urine in the montmorillonite 1 and 2 g/kg groups were lower compared with mice in the hyperuricaemic group. CONCLUSIONS: The results suggested that montmorillonite adsorbed uric acid and promoted diffusion of uric acid from blood vessels to intestine, prevented absorption of uric acid in intestine and decreased uric acid level in serum. PMID- 19903377 TI - Rubus sanctus protects against carbon tetrachloride-induced toxicity in rat isolated hepatocytes: isolation and characterization of its galloylated flavonoids. AB - OBJECTIVES: Rubus sanctus Schreb., known from the Bible as 'holy thorn bush', grows wild in Egypt. Rubus sanctus aqueous alcoholic extract (RE) contains a complicated phenolic mixture (ellagitanins, flavonoids and caffeic acid derivatives). In this study, the phytochemical investigation of the plant was re evaluated. Herein, we report on the isolation and identification of three galloylated flavonoids, namely kaempferol-3-O-(6''-O-galloyl)-(4)C(1)-beta-d galactopyranoside, quercetin-3-O-(6''-O-galloyl)-(4)C(1)-beta-d-galactopyranoside and myricetin-3-O-(6''-O-galloyl)-(4)C(1)-beta-d-galactopyranoside for the first time from the Rubus genus. We further aimed at evaluating the potential protective effects of RE against carbon tetrachloride (CCl(4))-induced toxicity in isolated rat hepatocytes. METHODS: Based on an initial concentration-response experiment, a concentration of 100 mug/ml was selected to investigate the hepatoprotective activity of RE. KEY FINDINGS: Pretreatment with RE afforded protection as indicated by counteracting CCl(4)-induced cell death, and reduced glutathione depletion. In addition, RE ameliorated CCl(4)-induced enzyme leakage by 40% for lactate dehydrogenase, 30% for alanine aminotransferase and 20% for aspartate aminotransferase as compared with CCl(4)-treated cells. Moreover, RE counteracted CCl(4)-induced lipid peroxidation and inhibited spontaneous lipid peroxidation in the control group. CONCLUSIONS: In conclusion, RE protects against CCl(4)-induced toxicity in isolated rat hepatocytes. PMID- 19903376 TI - Efficacy of different doses of aspirin in decreasing blood levels of inflammatory markers in patients with cardiovascular metabolic syndrome. AB - OBJECTIVES: Inflammation and platelet aggregation and activation are key processes in the initiation of a cardiovascular event. Patients with metabolic syndrome have a high risk of cardiovascular events. This study determined whether small and medium doses of aspirin have anti-inflammation and antiplatelet aggregation effects in patients with metabolic syndrome. METHODS: One hundred and twenty-one consecutive patients with metabolic syndrome were randomized into three groups, receiving 100 mg/day of aspirin, 300 mg/day of aspirin or a placebo, respectively, for 2 weeks. The blood levels of thromboxane B2 (TXB2), a stable product of the platelet aggregation mediator TXA2, 6-keto-prostaglandin F1 alpha (6-keto-PGF1-alpha), a stable product of the endogenous cyclooxygenase metabolite prostaglandin I2, and inflammatory mediators including high sensitivity C-reactive protein (hs-CRP), tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), were determined by ELISA and radioimmunoassay. KEY FINDINGS: The blood levels of hs-CRP, TNF-alpha, IL-6 and TXB2 were significantly decreased after 2 weeks of treatment with 300 mg/day of aspirin. Patients who received 100 mg/day of aspirin had decreased blood levels of hs-CRP and TXB2. The blood level of IL-6 in the 300 mg/day aspirin group was significantly lower than that in the other two groups after 2 weeks of therapy. Aspirin at either dose did not affect the blood level of 6-keto-PGF1-alpha. CONCLUSIONS: Aspirin at all doses suppresses the blood levels of inflammatory markers and the platelet aggregation mediator TXA2 in Chinese patients with metabolic syndrome. Since the suppression induced by 300 mg/day of aspirin was greater than that induced by 100 mg/day of aspirin, these data suggest that 300 mg/day of aspirin may be beneficial in decreasing the risk of cardiovascular events in Chinese patients with metabolic syndrome. PMID- 19903378 TI - Central nervous system effects of the essential oil of the leaves of Alpinia zerumbet in mice. AB - OBJECTIVES: Alpinia zerumbet, known in Brazil as colonia, is popularly used as a diuretic, antihypertensive, anti-ulcerogenic and sedative. Based on this, we have investigated the central effects of the essential oil isolated from A. zerumbet leaves. METHODS: Mice were treated once with 50 or 100 mg/kg of the essential oil, intraperitoneally, 30 min before being submitted to behavioural models of: locomotor activity (open-field), catalepsy, anxiety (elevated plus maze), depression (forced swimming test and tail suspension tests) as well as apomorphine-induced stereotypy. KEY FINDINGS: Results showed a dose-related decrease on locomotor activity and apomorphine-induced stereotypy. There was a decrease to the order of 55% of the grooming behaviour with both doses studied. The essential oil 100 mg/kg increased cataleptic activity (167%) and the immobility time in the forced swimming and tail suspension tests. Pretreatment with haloperidol (0.2 mg/kg, i.p.) alone also decreased locomotion, increased cataleptic activity and immobility time in the tail suspension test. No alterations in the elevated plus maze test were registered. CONCLUSIONS: The essential oil of A. zerumbet leaves had depressant and possible antipsychotic activity, since it could reverse the stereotypy induced by apomorphine, presenting effects comparable with those obtained with haloperidol treatment. PMID- 19903379 TI - Catecholamine-induced myocardial fibrosis and oxidative stress is attenuated by Terminalia arjuna (Roxb.). AB - OBJECTIVES: Myocardial fibrosis and oxidative stress accompany a number of cardiac disorders such as hypertrophic cardiomyopathy, hypertensive heart disease and cardiac failure. Stem bark of Terminalia arjuna has been advocated for cardiac ailments. The present study evaluated the effects of T. arjuna bark extract on myocardial fibrosis and oxidative stress induced by chronic beta adrenoceptor stimulation. METHODS: Aqueous extract of T. arjuna bark was evaluated at 63, 125 and 250 mg/kg given orally for antifibrotic and antioxidant effects in rats given the selective beta-adrenoceptor agonist isoprenaline (5 mg/kg s.c.) for 28 days. Captopril (50 mg/kg per day, given orally), an inhibitor of angiotensin-converting enzyme used as a standard cardioprotective drug, was used as a positive control. KEY FINDINGS: Isoprenaline caused fibrosis, increased oxidative stress and cardiac hypertrophy (increased heart weight : body weight ratio and cardiomyocyte diameter). The T. arjuna bark extract and captopril significantly prevented the isoprenaline-induced increase in oxidative stress and decline in endogenous antioxidant level. Both also prevented fibrosis but not the increase in heart weight : body weight ratio. CONCLUSIONS: T. arjuna protects against myocardial changes induced by chronic beta-adrenoceptor stimulation. PMID- 19903380 TI - Nootropic activity of lipid-based extract of Bacopa monniera Linn. compared with traditional preparation and extracts. AB - OBJECTIVES: The aim was to design an alternative solvent-free extraction method using the hydrophilic lipid Gelucire (polyethylene glycol glycerides) for herbal extraction and to confirm the efficacy of extraction using biological screening. METHODS: Bacopa monniera Linn. (BM) was selected for the study. Conventional methanolic extract (MEBM), Ayurvedic ghrita (AGBM) and lipid extracts (LEBM) were prepared and standardised by high-performance thin-layer chromatography (HPTLC). Nootropic activity in rats was evaluated using the two-trial Y-maze test and the anterograde amnesia induced by scopolamine (1 mg/kg i.p.) determined by the conditioned avoidance response. The extracts were administered daily at doses of 100, 200 and 400 mg/kg orally. At the end of the conditioned avoidance response test, brain monoamine levels were estimated by HPLC. KEY FINDINGS: The LEBM, MEBM and AGBM contained 3.56%, 4.10% and 0.005% bacoside A, respectively. Significantly greater spatial recognition was observed with LEBM (P < 0.001 at 400 and 200 mg/kg) and MEBM (P < 0.001 at 400 mg/kg, P < 0.01 at 200 mg/kg) than AGBM. The conditioned avoidance response was significantly higher in the groups treated with high doses of LEBM and MEBM than AGBM. There were significant decreases in brain noradrenaline (P < 0.001) and 5-hydroxytryptamine (P < 0.01) levels and an increase in dopamine levels (P < 0.05) in the LEBM-treated groups compared with the stress control group. CONCLUSIONS: The proposed LEBM is solvent free, does not have the shortcomings associated with conventional extraction, and had comparable nootropic activity to the MEBM. PMID- 19903381 TI - Raphanus sativus extract prevents and ameliorates zearalenone-induced peroxidative hepatic damage in Balb/c mice. AB - OBJECTIVES: Raphanus sativus (radish) is a species of crucifer, which includes widely consumed vegetables, distributed in Asia, Africa and Europe. It is a rich source of bioactive molecules including anthocyanins, glucosinolates, isothiocyanates and other flavonoids, and miscellaneous phenolic substances. We have evaluated the hepatoprotection of R. sativus extract against zearalenone, an estrogenic mycotoxin initiating hepatotoxicity in male Balb/c mice. METHODS: Animals were divided into seven treatment groups and treated orally each day for twenty eight days as follows: a control, an olive oil group, group I, group II, and group III treated with radish extract alone (5, 10 and 15 mg/kg, respectively), group IV treated with zearalenone (40 mg/kg), and group V treated with zearalenone plus the lowest dose of radish extract. KEY FINDINGS: Administration of zearalenone alone resulted in significant decreases in the levels of alkaline phosphatase, lactate dehydrogenase, alanine and aspartate aminotransferases in the liver, suggesting hepatic damage. Moreover, a marked increase in the level of lipid peroxide and concomitant decrease of glutathione peroxidase, glutathione reductase, superoxide dismutase, catalase, glutathione-S transferase, RNA and DNA concentrations were also observed in the liver tissue of zearalenone-treated mice. Co-treatment with R. sativus extract plus zearalenone succeeded in reversing the condition back to normal levels for all studied parameters. CONCLUSIONS: By itself R. sativus extract did not show any toxic effects and could be considered as a potent hepatoprotectant. PMID- 19903382 TI - The relation between moisture-induced aggregation and structural changes in lyophilized insulin. AB - OBJECTIVES: Long-term stability is a critical factor in the successful development of protein pharmaceuticals. Due to the relative instability of proteins in aqueous solutions, they are formulated frequently and stored as lyophilized powders. Exposure of such powders to moisture constitutes a substantial storage problem leading to aggregation and inactivation. We have investigated the structural consequences of moisture sorption by lyophilized insulin under controlled humidity conditions by employing Fourier transform infrared (FT-IR) microscopy. METHODS: Lyophilized insulin samples were stored in humidity chambers under controlled conditions at 50(o)C. Protein aggregation studies were carried out by redissolving the insulin samples and measuring the amount of both soluble protein and insoluble aggregates. Near-UV circular dichroism spectra were collected to assess the tertiary structure. FT-IR microscopy studies were carried out to investigate secondary structural changes in solid-state insulin after incubation at different relative humidities. KEY FINDINGS: It was found that sorption of moisture was accompanied by small structural changes in lyophilized insulin at low levels of relative humidity (i.e. 11%). At higher relative humidity levels, structural changes were becoming more pronounced and were characterized by a loss in the alpha-helix and increase in beta-sheet content. The magnitude of the structural changes in tendency paralleled the solid-state instability data (i.e. formation of buffer-insoluble aggregates and loss in tertiary structure upon reconstitution). CONCLUSIONS: The results support the hypothesis that water sorption by lyophilized proteins enables structural transitions which can lead to protein aggregation and other deleterious phenomena. PMID- 19903384 TI - Transfer of Bacillus cereus spores from packaging paper into food. AB - Food packaging papers are not sterile, as the manufacturing is an open process, and the raw materials contain bacteria. We modeled the potential transfer of the Bacillus cereus spores from packaging paper to food by using a green fluorescent protein-expressing construct of Bacillus thuringiensis Bt 407Cry(-) [pHT315Omega(papha3-gfp)], abbreviated BT-1. Paper (260 g m(-2)) containing BT-1 was manufactured with equipment that allowed fiber formation similar to that of full-scale manufactured paper. BT-1 adhered to pulp during papermaking and survived similar to an authentic B. cereus. Rice and chocolate were exposed to the BT-1-containing paper for 10 or 30 days at 40 or 20 degrees C at relative air humidity of 10 to 60%. The majority of the spores remained immobilized inside the fiber web; only 0.001 to 0.03% transferred to the foods. This amount is low compared with the process hygiene criteria and densities commonly found in food, and it does not endanger food safety. To measure this, we introduced BT-1 spores into the paper in densities of 100 to 1,000 times higher than the amounts of the B. cereus group bacteria found in commercial paper. Of BT-1 spores, 0.03 to 0.1% transferred from the paper to fresh agar surface within 5 min of contact, which is more than to food during 10 to 30 days of exposure. The findings indicate that transfer from paper to dry food is restricted to those microbes that are exposed on the paper surface and readily detectable with a contact agar method. PMID- 19903385 TI - Bacterial dynamics in model cheese systems, aiming at safety and quality of Portuguese-style traditional ewe's cheeses. AB - An experiment using model ewe's milk cheeses was designed to characterize microbial interactions that arise in actual raw milk cheese environments. These model cheeses were manufactured according to Portuguese artisanal practices, except that the microbial load and biodiversity were fully controlled: single potential pathogens and spoilage bacteria, or a combination thereof, were combined at various initial inoculum levels in sterilized raw ewe's milk with several lactic acid bacteria (LAB) normally found in traditional cheeses. Viable microbial counts were monitored throughout a 60-day ripening period. Two alternative mathematical approaches were used to fit the experimental data generated in terms of population dynamics: percent of inhibition and D-values. These were able to explain the complex competitive interactions between the contaminant microorganisms and the LAB adventitious populations. In general, the tested LAB were less able to inhibit contaminants present in combination and in higher concentrations. Lactococcus lactis, with its strong acidifying potential, was the most effective factor in controlling the unwanted bacterial population, especially single Staphylococcus aureus. The two lactobacilli studied, especially Lactobacillus brevis, were shown to be less effective; Escherichia coli and Listeria innocua were the contaminants least inhibited by the LAB. PMID- 19903383 TI - Protective effects of sodium para-amino salicylate on manganese-induced neuronal death: the involvement of reactive oxygen species. AB - OBJECTIVES: This study tested whether sodium para-amino salicylic dihydrate, an antibacterial drug for tuberculosis, could block manganese-induced apoptosis in SK-N-MC neurons. METHODS: Cell viability, Hoechst staining, dichlorofluorescin diacetate analysis for reactive oxygen species measurement, and immunoblotting were performed. KEY FINDINGS: In vitro, manganese chloride significantly decreased the viability of SK-N-MC cells, accompanied by apoptotic features such as changes in nuclear morphology. Sodium para-amino salicylic dihydrate inhibited these apoptotic characteristics through reducing intracellular reactive oxygen species generation, protecting mitochondrial membrane potential and caspase-3 activation. CONCLUSIONS: Sodium para-amino salicylic dihydrate inhibits manganese induced apoptosis in neurons and may reduce manganese-mediated neurodegeneration. PMID- 19903386 TI - Assessing interventions by quantitative risk assessment tools to reduce the risk of human salmonellosis from fresh minced pork meat in Belgium. AB - The risk of human salmonellosis through the consumption of minced pork meat in Belgium was assessed via a modular risk model covering pork meat production from lairage to human consumption. The main goal of the model was to give concrete options to reduce effectively the risk of human salmonellosis through the consumption of minced pork meat. These options (scenarios) were elaborated with reference to the international situation and the literature to give concrete and realistic possibilities for improving the microbiological quality of pork meat and to reduce the number of human salmonellosis cases per year in Belgium. The model estimates 15,376 cases of human salmonellosis per year in Belgium due to the consumption of minced pork meat. The results of the scenarios showed that the risk of human salmonellosis could be significantly reduced by efforts all along the pork meat production chain but also by efforts made by consumers. The responsibility of food business operators for the pork meat production chain is high in relation to the microbiological quality of meat delivery, especially at the slaughterhouse. Consumers also need to be aware of good hygiene practices during preparation of the meat at home. Cross-contamination with raw food can be avoided by changing the habits and the behavior of the household cook. The results of these scenarios would be useful for the food business operators involved in the pork meat chain and for public health authorities. PMID- 19903387 TI - Assuring growth inhibition of listerial contamination during processing and storage of traditional Greek Graviera cheese: compliance with the new European Union regulatory criteria for Listeria monocytogenes. AB - The current microbiological regulatory criteria in the European Union specify a maximum Listeria monocytogenes population of 100 CFU/g allowable in ready-to-eat foods provided the product will not exceed this limit throughout its shelf life. The aim of this study was to validate the manufacturing method for traditional Greek Graviera cheese produced from thermized milk. Initial challenge experiments evaluated the fate of inoculated L. monocytogenes (ca. 4 log CFU/ml, three-strain cocktail) in thermized Graviera cheese milk (TGCM; 63 degrees C for 30 s) in the presence and absence of a product-specific starter culture (SC) in vitro. Milk samples were incubated for 6 h at 37 degrees C and then for 66 h at 18 degrees C. Experiments were conducted to evaluate the fate of a cocktail of three nonpathogenic L. monocytogenes and L. innocua indicator strains inoculated (ca. 3 log CFU/g) in Graviera cheeses commercially manufactured from TGCM+SC. Cheeses were brined, ripened at 18 degrees C and 90% relative humidity for 20 days, and stored at 4 degrees C for up to day 60 under vacuum. In TGCM, L. monocytogenes increased by ca. 2 log units, whereas in TGCM+SC L. monocytogenes growth was retarded (P < 0.05) after a ca. 1-log increase within 6 h at 37 degrees C. Populations of Listeria indicator strains did not grow in TGCM+SC cheeses at any stage; they declined 10-fold in fresh cheeses within 5 days and then survived with little death thereafter. Thus, growth inhibition but not inactivation of potent natural Listeria contaminants at levels below 100 CFU/g occurs in the core of traditional Greek Graviera cheese during fermentation, ripening, and storage. PMID- 19903388 TI - Efficacy of chlorine dioxide against Listeria monocytogenes in brine chilling solutions. AB - Chilled brine solutions are used by the food industry to rapidly cool ready-to eat meat products after cooking and before packaging. Chlorine dioxide (ClO(2)) was investigated as an antimicrobial additive to eliminate Listeria monocytogenes. Several experiments were performed using brine solutions made of sodium chloride (NaCl) and calcium chloride (CaCl(2)) inoculated with L. monocytogenes and/or treated with 3 ppm of ClO(2). First, 10 and 20% CaCl(2) and NaCl solutions (pH 7.0) were inoculated with a five-strain cocktail of L. monocytogenes to obtain approximately 7 log CFU/ml and incubated 8 h at 0 degrees C. The results demonstrated that L. monocytogenes survived in 10% CaCl(2), 10 and 20% NaCl, and pure water. L. monocytogenes levels were reduced approximately 1.2 log CFU/ml in 20% CaCl(2). Second, inoculated ( approximately 7 log CFU/ml) brine solutions (10 and 20% NaCl and 10% CaCl(2)) treated with 3 ppm of ClO(2) resulted in a approximately 4-log reduction of the pathogen within 90 s. The same was not observed in a solution of 20% CaCl(2); further investigation demonstrated that high levels of divalent cations interfere with the disinfectant. Spent brine solutions from hot dog and ham chilling were treated with ClO(2) at concentrations of 3 or 30 ppm. At these concentrations, ClO(2) did not reduce L. monocytogenes. Removal of divalent cations and organic material in brine solutions prior to disinfection with ClO(2) should be investigated to improve the efficacy of the compound against L. monocytogenes. The information from this study may be useful to processing establishments and researchers who are investigating antimicrobials in chilling brine solutions. PMID- 19903389 TI - Spoilage and safety characteristics of ground beef treated with lactic acid bacteria. AB - Lactic acid bacteria (LAB) can decrease numbers of Escherichia coli O157:H7 and Salmonella in ground beef during storage. Two dose-titration studies were conducted in ground beef to determine dose levels of LAB needed to inhibit the pathogens. A second study evaluated whether LAB masked changes typically associated with the spoilage of ground beef displayed under refrigerated (0 degrees C) or abusive (10 degrees C) temperatures packaged in both traditional overwrap (TOP) and modified atmosphere packaging (MAP; 80% O(2)-20% CO(2)). Microbial analyses were conducted to determine spoilage endpoints and pathogen reduction. In the dose-titration study, Salmonella was reduced by 3 log cycles at all doses (10(6), 10(7), and 10(8) LAB per g) after 3 days of storage and was eliminated after 5 days of storage. E. coli O157:H7 was reduced by 2 log cycles at all dosages after 3 days of storage and by 3 log cycles after 5 days of storage. In the spoilage studies, as expected, total aerobic plate counts and LAB populations in LAB-inoculated samples were higher than the controls initially, but the counts were similar near the end of the study. While total spoilage bacteria generally increased over time, very few differences existed between treatments stored at 0 degrees C and 10 degrees C in coliforms, Brochothrix thermosphacta, yeasts and molds, and Pseudomonas spp. counts for both the TOP and MAP samples. We conclude that LAB could potentially be added to ground beef in TOP and MAP as a processing intervention for E. coli O157:H7 and Salmonella without masking microbial spoilage characteristics. PMID- 19903390 TI - Evaluation of a biocontrol preparation consisting of Enterobacter asburiae JX1 and a lytic bacteriophage cocktail to suppress the growth of Salmonella Javiana associated with tomatoes. AB - A biocontrol preparation based on a combination of Enterobacter asburiae JX1 and a cocktail of five lytic bacteriophages was evaluated for control of Salmonella Javiana within the rhizosphere of plants and in pre- and postharvest tomatoes. The biocontrol preparation introduced into the rhizosphere of growing tomato plants reduced the persistence of Salmonella, although no synergistic action was observed between E. asburiae JX1 or the bacteriophage cocktail when used in combination. When the biocontrol preparation was coinoculated with Salmonella onto the blossom of tomato plants, the prevalence of the enteric pathogen both on the surface and in internal tissues of the subsequent tomatoes was significantly reduced compared with controls. Tomatoes derived from plants inoculated with Salmonella alone had a prevalence of 92% surface contamination (22 of 24 tomato batches were positive for Salmonella) and 43% internal contamination (31 of 72 batches positive). This Salmonella prevalence was reduced to 0% (0 of 38 positive) and 2% (1 of 57 positive), respectively, when the biocontrol preparation was applied. Although bacteriophages reduced the prevalence of internalized Salmonella, the main growth suppressing effect was via the antagonistic activity of E. asburiae JX1. No bacteriophages were recovered from tomatoes despite being introduced at 6 log PFU onto the blossom of plants. The biocontrol preparation was not effective for controlling the growth of Salmonella introduced onto postharvest tomatoes that were stored for 7 days at 15 degrees C. The application of E. asburiae JX1 is a promising approach for controlling Salmonella encountered in tomato production, and there was no evidence to suggest that the antagonistic activity could be enhanced by the coinoculation of bacteriophages. PMID- 19903391 TI - Morphology, release characteristics, and antimicrobial effect of nisin-loaded electrospun gelatin fiber mat. AB - Gelatin electrospun (e-spun) fiber mats containing nisin were produced by electrostatic spinning of gelatin-nisin in 70% (vol/vol) acetic acid aqueous solutions. Varying nisin loading concentration (0 to 3% [wt/wt]) did not affect the fiber average diameter, whereas increasing gelatin concentration from 20 to 24% (wt/vol) caused an increase in the average diameter. All nisin-loaded gelatin e-spun fiber mats demonstrated inhibition against Lactobacillus plantarum TISTR 850. However, all fiber mats were fragile and easily dissolved in water. Cross linking by saturated glutaraldehyde vapor at 37 degrees C for 5 min was done to strengthen the mat. Tensile strength, Young's modulus, and elongation of the cross-linked gelatin-nisin e-spun fiber mats varied in the range of 2.6 to 20.3 MPa, 163 to 966 MPa, and 1.7 to 5.9% , respectively. Cross-linking did not affect the mat's inhibition activity against L. plantarum TISTR 850. Nisin retention in cross-linked antimicrobial gelatin e-spun fiber mats was in the range of 1.0 to 1.22% . Increasing temperature caused an increase in nisin release, but increasing water activity did not cause a significant difference in nisin release over 50 h. After storage at 25 degrees C for 5 months, the antimicrobial gelatin e-spun fiber mat still showed inhibition against L. plantarum TISTR 850. The mats also inhibited the growth of Staphylococcus aureus and Listeria monocytogenes but not Salmonella Typhimurium. PMID- 19903392 TI - Growth response of Escherichia coli ATCC 35218 adapted to several concentrations of sodium benzoate and potassium sorbate. AB - Escherichia coli ATCC 35218 growth response was evaluated after repetitive cultivation in stepwise increasing antimicrobial agent concentrations (potassium sorbate or sodium benzoate) to observe its adaptation process to high weak-acid concentrations. The effect of antimicrobial (potassium sorbate or sodium benzoate) concentration (0 to 7,000 ppm) was tested using laboratory media. Cells adapted at 1,000 ppm were inoculated in media containing the same concentration of the antimicrobial; after that, cells were transferred to media containing a higher concentration, followed by repetitive cultivations. In every case, viable cells were determined by surface plating every hour up to 48 h. Logarithmic representations of survival or growing fraction were modeled using the Gompertz equation. Adapted and nonadapted cells were analyzed for plasmid presence as well as phosphofructokinase and succinate dehydrogenase activity. Bacterial growth was observed after adaptation processes in media formulated up to 7,000 ppm of potassium sorbate or sodium benzoate. Analyses of variance demonstrated that no significant difference (P > 0.05) in lag time or growth rate was observed among adapted cells cultured in media containing the studied concentrations for each of the antimicrobials tested. These results suggest that E. coli can be adapted to high weak-acid concentrations if the exposure is performed under sublethal conditions. Furthermore, there was demonstrated inhibition of the enzymes phosphofructokinase and succinate dehydrogenase by action of sodium benzoate and potassium sorbate, respectively. E. coli adaptation to antimicrobial agents was not related to plasmid presence but appears to be due to other action mechanisms. PMID- 19903393 TI - Transfer of Escherichia coli O157:H7 from soil, water, and manure contaminated with low numbers of the pathogen to lettuce plants. AB - The sources of contamination of leafy greens remain unclear, but it is evident that contaminated water, soil amendments, and wildlife likely contribute. The objective of the present study was to determine transfer of low numbers of Escherichia coli O157:H7 from soil, manure-amended soil, and water to growing lettuce plants. Lettuce plants, young (12 days of age) or mature (30 days of age), were grown in soil, manure-amended soil, or irrigated with water containing 10(1), 10(2), 10(3), or 10(4) CFU E. coli O157:H7 per g or ml. Harvested plants were processed to determine whether E. coli O157:H7 was associated with the entire plant or within internal locations. Young plants (12 days) were harvested at 1, 10, 20, and 30 days postexposure. No samples were positive for E. coli O157:H7 after direct plating of serial dilutions. Enrichment of all samples from young plants exposed to contaminated soil, manure-amended soil, and irrigation water demonstrated that approximately 21% (113 of 552) of plants were positive for E. coli O157:H7. Approximately 30% (36 of 120) of the mature plants initially irrigated with or grown in contaminated soil (including manure-amended soil) for 15 days were positive for E. coli O157:H7. Based on sterilization of surface tissue, E. coli O157:H7 was in protected locations of lettuce tissue. The results suggest that lettuce exposed to, and grown in the presence of, low numbers of E. coli O157:H7 may become contaminated and thus present a human health risk. PMID- 19903394 TI - Effects of plant maturity and growth media bacterial inoculum level on the surface contamination and internalization of Escherichia coli O157:H7 in growing spinach leaves. AB - The incidence of foodborne outbreaks linked to fresh produce has increased in the United States. Particularly noteworthy was the 2006 Escherichia coli O157:H7 outbreak associated with prepackaged baby spinach. This study aimed to determine whether E. coli O157:H7 would be present in the aerial leaf tissue of a growing spinach plant when introduced at various plant maturities and different inoculum levels in a greenhouse setting. Spinach seeds of a commercial variety were sown in 8-in. (20.32-cm) pots. After seed germination, two levels (10(3) and 10(7) CFU/ml) of an E. coli O157:H7 green fluorescent protein-expressing strain were introduced into the plant growth media weekly for a total of five times. Inoculated spinach plants were examined weekly for the presence of E. coli O157:H7 on leaves and in surrounding growth media. Among 120 spinach plant samples examined for internal leaf contamination, only one yielded a positive result. Surface leaf contamination occurred occasionally and clustered between 3 and 5 weeks of age, but not among leaves younger than 3 weeks of age. On the other hand, when inoculated at the 10(7) CFU/ml level, the E. coli O157:H7 green fluorescent protein strain survived the entire cultivation period, although with gradually reduced levels. The experiments demonstrated that internalization of E. coli O157:H7 of growing spinach plant leaves under greenhouse conditions was a rare event, but surface contamination did occur, primarily when the plants reached 3 weeks of age. The study provided important data to further assess the association between spinach age and potential contamination of E. coli O157:H7. PMID- 19903395 TI - Impact of organic and conventional management on the phyllosphere microbial ecology of an apple crop. AB - Bacterial communities associated with the phyllosphere of apple trees (Malus domestica cv. Enterprise) grown under organic and conventional management were assessed to determine if increased biological food safety risks might be linked with the bacterial communities associated with either treatment. Libraries of 16S rRNA genes were generated from phyllosphere DNA extracted from a wash made from the surfaces of leaves and apples from replicated organic and conventional treatments. 16S rRNA gene libraries were analyzed with software designed to identify statistically significant differences between bacterial communities as well as shared and unique phylotypes. The identified diversity spanned eight bacterial phyla and 14 classes in the pooled organic and conventional libraries. Significant differences between organic and conventional communities were observed at four of six time points (P < 0.05). Despite the identification of significantly diverse microfloras associated with organic and conventional treatments, no detectable differences in the presence of potential enteric pathogens could be associated with either organic or conventional management. Neither of the bacterial genera most commonly associated with produce-related illness outbreaks (Salmonella and Escherichia) was observed in any of the libraries. The impressive bacterial diversity that was documented in this study provides a valuable contribution to our developing understanding of the total microbial ecology associated with the preharvest phyllospheres of food crops. The fact that organic and conventional phyllosphere bacterial communities were significantly different at numerous time points suggests that crop management methods may influence the bacterial consortia associated with the surfaces of fruits and vegetables. PMID- 19903396 TI - Risk factors at slaughter associated with presence of Salmonella on hog carcasses in Canada. AB - Despite the application of hazard analysis and critical control point systems at slaughter and during processing, Salmonella contamination is still a significant biological hazard associated with pork products. A better understanding of risk factors in slaughterhouses and of contamination sources is therefore critical to improve control of this bacterium in the abattoirs. The objectives of this study were to identify the risk factors at slaughter that are associated with the presence of Salmonella on hog carcasses and to assess possible sources of contamination. A questionnaire on potential risk factors was developed. Over 7,400 hogs originating from 312 randomly selected production lots were tested. The lots were from 10 different abattoirs located in five different Canadian provinces. At slaughter, blood was collected for serological analysis, and mesenteric lymph nodes (MLN) and carcass swabs were collected for Salmonella analysis. Furthermore, pulsed-field gel electrophoresis was conducted to establish the genetic profiles of selected isolates from carcasses and MLN and to compare these profiles with those recovered from the slaughter environment. Multivariate regression analysis results indicated that the cleanliness of the hogs and the status of the scald water were factors significantly associated with the Salmonella status of the carcasses at the end of the slaughter process. Pulsed-field gel electrophoresis analysis showed that most isolates from carcasses were similar to those from animals (MLN) or the preevisceration environment. PMID- 19903397 TI - Restaurant Salmonella Enteritidis outbreak associated with an asymptomatic infected food worker. AB - Salmonella is the most common bacterial cause of foodborne outbreaks in the United States; approximately half of Salmonella outbreaks occur in restaurant settings. In February 2008, investigation of a cluster of Salmonella Enteritidis cases with indistinguishable pulsed-field gel electrophoresis (PFGE) patterns revealed that five cases had eaten at the same restaurant. Cases were identified through routine surveillance activities and by contacting meal companions of culture-confirmed cases. Well meal companions and well patrons contacted via check stubs served as controls. Illness histories and stool samples were collected from all restaurant employees. Sandwiches were the only menu item or ingredient significantly associated with illness (15 of 15 cases versus 17 of 37 controls; odds ratio, undefined; P < 0.001). None of the six restaurant employees reported experiencing recent gastrointestinal symptoms. The outbreak PFGE subtype of Salmonella Enteritidis was identified in two food workers. One of the positive employees began working at the restaurant shortly before the first exposure date reported by a case, and assisted in the preparation of sandwiches and other foods consumed by cases. The other positive employee rarely, if ever, handled food. The restaurant did not have a glove use policy. There was no evidence of ongoing transmission after exclusion of the positive food workers. This was a restaurant Salmonella Enteritidis outbreak associated with an asymptomatic infected food worker. Routine PFGE subtyping of Salmonella Enteritidis isolates, routine interviewing of cases, and an iterative approach to cluster investigations allowed for timely identification of the source of an outbreak of Salmonella Enteritidis infections. PMID- 19903398 TI - Prevalence and molecular diversity of Listeria monocytogenes in retail establishments. AB - As our understanding of Listeria monocytogenes transmission in retail and deli operations is limited, we conducted a cross-sectional study of L. monocytogenes contamination patterns in 121 retail establishments, using testing of food and environmental samples and subtype analysis (ribotyping) of L. monocytogenes isolates. Seventy-three (60%) establishments had at least one sample that tested positive for L. monocytogenes; 5 (2.7%) of the 183 food and 151 (13.0%) of the 1,161 environmental samples tested positive for L. monocytogenes, including 125 (16.7%) and 26 (6.3%) of non-food contact and food contact surface samples, respectively. Thirty-two EcoRI ribotypes were identified among the 156 L. monocytogenes isolated. Twenty-seven establishments had two or more L. monocytogenes with the same ribotype within a given establishment, including 9 establishments where isolates from 3 to 5 samples had the same ribotype. In 5 of 7 establishments where follow-up sampling was conducted 8 to 19 months after the initial sampling, isolates with the same ribotype were obtained in both samplings; persistence of a given strain was also confirmed by pulsed-field gel electrophoresis. Our data indicate that (i) L. monocytogenes is regularly found in some retail environments; (ii) L. monocytogenes strains are often widely distributed in retail, indicating cross-contamination and dispersal; (iii) L. monocytogenes can persist in retail environments for more than 1 year; and (iv) a number of L. monocytogenes subtypes isolated at retail are common among human listeriosis cases. We also identified specific contamination patterns in retail establishments, providing critical information for the development of L. monocytogenes control strategies. PMID- 19903399 TI - Sensitive and rapid molecular detection assays for Salmonella enterica serovars Typhimurium and Heidelberg. AB - Salmonella enterica is a significant cause of gastroenteritis worldwide, with serovars Typhimurium and Heidelberg being particularly prevalent, which have broad host ranges infecting poultry, dairy animals, and humans. Traditional methods used for the detection of Salmonella from contaminated food products are time-consuming and labor-intensive. The aim of this study was to develop a sensitive and rapid PCR-based detection method with optimized specificity for high-throughput screening of food and clinical samples. We used bioinformatics to identify potential serovar-specific regions from the available S. enterica sequenced genomes. We designed primer pairs to targeted regions unique to Typhimurium and Heidelberg. A primer pair targeting a putative cytoplasmic protein STM4492 amplified a 759-bp product specific to Typhimurium, and a primer pair targeting a putative inner membrane protein STM2745 amplified a 199-bp product from both Typhimurium and Heidelberg. A primer pair for the oriC locus was used to identify all Salmonella. We screened 217 isolates including the Salmonella reference collections A and B, validating the specificity of each primer set. Next, a multiplex PCR (mPCR) assay and quantitative real-time PCR assay were optimized for identification and differentiation of Typhimurium and Heidelberg. An mPCR assay was developed and successfully detected S. enterica isolates from inoculated Cheddar cheese, raw turkey, and cooked turkey at concentrations as low as 1 CFU/g of food. The reaction conditions for this mPCR have significantly reduced the time needed to identify S. enterica Typhimurium and Heidelberg, making this a rapid selective tool. PMID- 19903400 TI - Use of FRNA bacteriophages to indicate the risk of norovirus contamination in Irish oysters. AB - Male-specific (F) RNA bacteriophages have been proposed as indicators for human enteric viruses in shellfish. This study compared the use of Escherichia coli and FRNA bacteriophages to indicate the presence and level of noroviruses in Crassostrea gigas. A total of 167 samples from category A and B shellfish harvesting areas were analyzed for E. coli and FRNA bacteriophages by standard methods and for noroviruses (NoV) by using a previously described real-time PCR assay. FRNA bacteriophage and NoV levels in shellfish showed a seasonal trend and were elevated during the winter period (October through March). Conversely, E. coli levels did not reflect this seasonal trend. Categorizing samples on the basis of E. coli levels according to European Union regulatory limits failed to indicate the occurrence or level of NoV in shellfish. However, by grouping shellfish samples on the basis of FRNA bacteriophage levels a clear correlation was observed with NoV levels. The use of FRNA bacteriophages to predict the occurrence of NoV in shellfish could provide improved public health protection and should be considered when developing risk management procedures for shellfisheries. PMID- 19903401 TI - Neonatal mice as models for Cronobacter sakazakii infection in infants. AB - Cronobacter sakazakii is an opportunistic pathogen that has been isolated from powdered infant formulas. C. sakazakii infection can result in serious illnesses such as bacteremia, septicemia, meningitis, and death in at-risk infants who are orally fed contaminated reconstituted powdered infant formulas. The objective of this study was to compare the susceptibilities of BALB/c, C57BL/6, and CD-1 mice to C. sakazakii strain MNW2. We acquired timed-pregnant CD-1 mice and allowed them to give birth naturally. On postnatal day 3.5, each pup was administered a total dose of approximately 10(2) to 10(11) CFU C. sakazakii strain MNW2 in reconstituted powdered infant formula. Mice were observed twice daily for morbidity and mortality. At postnatal day 10.5, the remaining pups were euthanized, and brain, liver, and cecum were excised and analyzed for the presence of C. sakazakii. C. sakazakii was isolated from brains, livers, and ceca in all three mouse strains. The CD-1 mouse strain was the most susceptible of the three, with the lowest infectious dose (10(2) CFU) and the lowest lethal dose (also 10(2) CFU). PMID- 19903402 TI - Development, evaluation, and peer verification of a rapid real-time PCR method for the detection of animal material. AB - Four real-time PCR assays that can be used with U.S.- and European Union-rendered materials to detect three ruminant species (bovine, caprine, and ovine) and a select set of avians (chicken, goose, and turkey) were developed. This method was evaluated against stringent acceptance criteria previously developed by the U.S. Food and Drug Administration, Center for Veterinary Medicine's Office of Research. Acceptance criteria for determining success used a statistical approach requiring a 90% probability of achieving the correct response, within a 95% confidence interval. A minimum detection level of 0.1% meat and bone meal (MBM) was required, consistent with the sensitivity of the validated PCR-based method currently used by the U.S. Food and Drug Administration as an aid in enforcement of the Agency's feed ban. PCR primer specificity was determined by using a panel of DNA samples derived from 16 different animal species. The method is able to detect 0.1% rendered material in complete feed in less than 1.5 h of total assay time, a significant improvement over the current method, which requires 7 to 8 h for completion. The real-time assay for the detection of animal material passed stringent acceptance criteria for sensitivity, selectivity, and specificity. The method also passed ruggedness, real-time platform, and second analyst trials. Two external laboratories participating in a peer-verification trial demonstrated 100% specificity in identifying bovine MBM, ovine MBM, or caprine meat meal, while exhibiting a 0.6% rate of false positives. These results demonstrated that this method was capable of being used by other laboratories. PMID- 19903403 TI - Species identification of ciguatoxin-carrying grouper implicated in food poisoning. AB - Food poisoning due to ingestion of an unknown red grouper occurred in southern Taiwan. To identify the species of toxic red grouper implicated in food poisoning, a 475-bp sequence of the cytochrome b gene from six species of fresh red grouper meat was amplified by using a pair of primers (L14735/H15149). This fragment could be amplified when fish meat was treated with different heating processes. After sequencing, it was found that no variation in sequences was detected among individuals within each species. The species of toxic red grouper meat implicated in food poisoning was judged to be Lutjanus bohar based on sequence analysis. In addition, restriction enzyme analysis with HaeIII rapidly distinguished these six species of red grouper and the two samples implicated in food poisoning. No toxicity of viscera in 18 specimens of six red grouper species was detected, but two food poisoning samples were found to be toxic. This study indicated that DNA sequence and restriction enzyme analysis are powerful methods for identifying potentially toxic red grouper species as L. bohar. PMID- 19903404 TI - Effects of temperature and pH on the growth of bacteria isolated from blown packs of vacuum-packaged beef. AB - Bacteria recovered from the microflora of blown packs of vacuum-packaged beef were identified as Leuconostoc mesenteroides, Lactococcus lactis, Carnobacterium maltaromaticum, and Clostridium estertheticum, with L. mesenteroides predominant. Isolates of these lactic acid bacteria all grew in peptone yeast extract glucose starch broth (PYGSB) at temperatures between -2 and 30 degrees C but generally grew more slowly and over a more restricted temperature range in meat juice medium (MJM). A C. estertheticum isolate and the type strain of C. estertheticum subsp. estertheticum (ATCC 51377) both grew in PYGSB and MJM at similar rates at temperatures between -2 and 17 degrees C and grew at 20 degrees C in MJM but not in PYGSB. Square root models of the variation of the growth rate with temperature indicated that the C. maltaromaticum isolate and the C. estertheticum strains grew at similar rates that were faster than those of the other isolates at temperatures between -2 and 0 degrees C. The L. mesenteroides and L. lactis isolates grew in PYGSB at pH 5.0, but the C. maltaromaticum isolate and both strains of C. estertheticum did not grow in PYGSB at pH 1,000-fold more in rice suspension than in cooked meat medium or phosphate buffer. It was therefore suggested that rice contains component(s) that facilitates thermal inactivation of C. botulinum. PMID- 19903409 TI - Leakage of intracellular UV materials of high hydrostatic pressure-injured Escherichia coli O157:H7 strains in tomato juice. AB - The behavior of high hydrostatic pressure-injured Escherichia coli O157:H7 cells (strain SEA13B88 and a strain from the June-July 1999 Oklahoma juice outbreak) in tomato juice (pH 4.1) and phosphate-buffered saline (PBS; pH 7.2) at final concentrations of 8.4 to 8.8 log CFU/ml, respectively, and treated at 400, 500, and 600 MPa for 40 min at 25 and 35 degrees C with storage at 5 and 23 degrees C for 1,800 min was investigated. Immediately after treatment and every 3 h for 24 h of storage, an aliquot (0.1 ml) was plated on Trypticase soy agar and sorbitol MacConkey agar to determine the percentage of injured population. Leakage of UV materials and possible recovery from injury were investigated. Pressure (600-MPa) treatment at 35 degrees C for 40 min caused a higher percentage of bacterial injury than for 10 min of treatment. A higher percentage of injured population was found among the Oklahoma strain cells than among strain SEA13B88 cells, and differences in viability loss for bacterial strains were determined. The viability loss determined in PBS was 4.8 log for SEA13B88 cells and 5.2 log for Oklahoma cells, while losses of 5.4 and 5.7 log were determined in tomato juice for SEA13B88 and Oklahoma cells, respectively. The leakage of intracellular materials of injured Oklahoma cells was higher than that observed for SEA13B88 cells, but injured Oklahoma cells recovered faster in PBS. However, injured and healthy populations for both strains were below detection in tomato juice stored at 5 degrees C for 1,440 min. PMID- 19903410 TI - A comparative study assaying commonly used sanitizers for antimicrobial activity against indicator bacteria and a Salmonella Typhimurium strain on fresh produce. AB - With increased concerns over failures in vegetable and fruit sanitation, evaluating the efficacy of widely approved chemicals is ever more important. The purpose of this study was to determine whether sanitation treatments are equally effective against indicator bacteria and human enteric pathogens on cucumber and parsley. We provide here an experimental overview on the efficacy of common sanitation methods, which are based on peracetic acid-hydrogen peroxide, sodium dichloroisocyanurate, and the quaternary ammonium compound didecyldimethylammonium chloride. The sanitizers were tested for their activity against natural populations of total aerobic microorganisms, enterococci, and coliforms, and against the enteric pathogen Salmonella Typhimurium ATCC 14028 (which was added artificially). Results revealed that compared with washing parsley and cucumbers with water, treatments with all three sanitizers were not effective, resulting in a maximal reduction of only 0.7 log CFU of Salmonella Typhimurium. These sanitizers were also not effective in removal of natural bacteria from parsley (maximal reduction was 0.7 log CFU). Sanitation of cucumber was more successful; peracetic acid showed the most effective result, with a reduction of 2.7 log in aerobic microorganisms compared with cucumbers washed with water. Still, removal of natural bacteria from cucumbers proved more efficient than the removal of Salmonella Typhimurium. This may create a debate about the necessity of the sanitation and its contribution to safety, because sanitation of some contaminated vegetables may result in an increased likelihood of foods that, although they are given good hygienic ratings due to low microbial counts, harbor pathogens. PMID- 19903411 TI - High-pressure homogenization for the inactivation of human enteric virus surrogates. AB - Novel inactivation methods are needed to control the spread of foodborne viruses responsible for nonbacterial gastroenteritis worldwide. The advent of high pressure homogenization combining high pressure, shear stress, and cavitation provides the opportunity to evaluate this technology for viral inactivation in fluid foods under continuous processing conditions. Our objective was to evaluate murine norovirus (MNV-1) and MS2 coliphage (single-stranded RNA) as human enteric virus surrogates for their susceptibility to a novel high-pressure homogenization process for application in commercial settings. Experiments were conducted in duplicate with MNV-1 and MS2 coliphage in phosphate-buffered saline, using homogenization pressures of 0, 100, 200, 250, and 300 MPa (the maximum achievable by the homogenizer), resulting in exposure temperatures of 24, 46, 63, 70, and 75 degrees C, respectively, for <2 s. Only homogenization pressures of 300 MPa at 75 degrees C showed inactivation of approximately 3 log PFU for MS2 from an initial approximately 6 log PFU. Also, MNV-1 showed inactivation of approximately 0.8 log PFU at 300 MPa. Further studies are warranted to validate this inactivation process, which can retain the sensory and nutritional value of fluid food and shows promise for application in industrial environments. PMID- 19903412 TI - Prevalence and antimicrobial resistance of Salmonella isolated from broiler farms, chicken carcasses, and street-vended restaurants in Casamance, Senegal. AB - This study was undertaken to determine the prevalence and distribution of Salmonella on 57 randomly selected broiler farms at the end of the rearing period and in chicken products in urban and periurban areas in Casamance, Senegal, and to evaluate the antimicrobial resistance profiles of the Salmonella serovars. Salmonella was detected in chicken feces, on carcass skin, and in muscle on 35.1, 38.6, and 29.8% of farms, respectively. Salmonella was found in chicken meat servings from 14.3% of the 42 street restaurants and in 40.4% of the 285 chicken carcasses examined. The prevalence on skin and in muscle was significantly associated with the detection of Salmonella in feces (P 0.05) to the results found with traditional plating media for naturally contaminated broiler meat, regardless of whether the comparison was made between B+B and B-B, or among different meat products (breast, tenders, and thighs). More positive samples were found at 48 h of enrichment than at 24 h of enrichment (P < 0.05). A Campylobacter jejuni:Campylobacter coli ratio of 4:1 was found in this study. Most of the isolates from both subsamples (B+B and B-B) were similar or identical by PFGE analysis, except for a few samples in which the PFGE profiles of the isolates from the subsamples were different. Mini VIDAS allowed for the detection of Campylobacter spp. within 48 h after enrichment. However, the sensitivity is similar to plate media, and retail broiler samples need to be enriched for 48 h to avoid false negatives. PMID- 19903414 TI - Detection of allergen walnut component in food by an improved real-time PCR method. AB - A real-time PCR method aimed at the gene sequence of the walnut vicilin-like seed storage protein was established for the detection of the allergen walnut in food. The primers and probe were designed based on published methods. The method provided positive results for walnut and negative results for other tested agricultural plant materials including pecan. The intrinsic detection limit of the method was 0.00125 ng of walnut DNA, and the practical detection limit was 0.001% (wt/wt) walnut content in wheat; both of these values are lower than that of previously published methods. Therefore, this real-time PCR method is sufficiently specific and sensitive for the detection of walnut component in food. PMID- 19903415 TI - Clinical value of computed tomography perfusion source images in acute stroke. AB - Computed tomography perfusion (CTP) map can sensitively and accurately distinguish between infarct core and ischemic penumbra. However, CTP mapping software might not generate a perfusion map because of head movement; thus, analysing CTP source images (CTP-SI) is necessary in this situation to provide information for stroke diagnosis and therapy. In our work, 'one-stop shop' computed tomography (CT) examination including non-contrast-enhanced CT (NCCT), CTP, CT angiography (CTA) were performed in 24 patients with symptoms of acute stroke less than 9 hours. We divided patients into two groups (with and without delayed perfusion on CTP-SI), and compared the Alberta Stroke Program Early CT Score (ASPECTS) on CTP-SI and CTA-SI with follow-up imaging. Using follow-up imaging ASPECTS as the final infarct size, our results suggests that the ASPECTS of both CTP-SI and CTA-SI effectively predict final infarct core in the group without delayed perfusion, whereas CTP-SI has a potential advantage over CTA-SI in being able to predict final infarct core in the group with delayed perfusion. In conclusion, CTP-SI provides useful complementary information when CTP map software could not generate perfusion maps. PMID- 19903416 TI - The clinical effectiveness of glucosamine and chondroitin supplements in slowing or arresting progression of osteoarthritis of the knee: a systematic review and economic evaluation. AB - OBJECTIVE: To assess the clinical effectiveness and cost-effectiveness of glucosamine sulphate/hydrochloride and chondroitin sulphate in modifying the progression of osteoarthritis (OA) of the knee. DATA SOURCES: Electronic databases were searched from 1950 to 2008 and included: MEDLINE and PubMed; EMBASE; Cochrane Library (including Cochrane Systematic Reviews Database, CENTRAL, DARE, NHS EED and HTA databases); Allied and Complementary Medicine (AMED); National Research Register (NRR); Web of Science Proceedings; Current Controlled Trials; and Clinical Trials.gov. Other sources included bibliographies of retrieved papers, registered but unpublished trials, internet searches and the Food Standards Agency website. REVIEW METHODS: A search was conducted for systematic reviews of randomised controlled trials (RCTs), which were used to identify RCTs of at least 12 months' duration and updated with searches for primary studies. A cost-effectiveness model was constructed using cohort simulation and drawing on available evidence. Sensitivity analysis was undertaken and value of information analysis conducted. A review of studies of mechanism of action was carried out to explore the biological plausibility of the preparations. RESULTS: Five systematic reviews and one clinical guideline met the inclusion criteria. They reported inconsistent conclusions with only modest effects on reported pain and function. A reduction in joint space narrowing was more consistently observed, but the effect size was small and the clinical significance uncertain. A separate review of eight primary trials of > 12 months' duration showed evidence of statistically significant improvements in joint space loss, pain and function for glucosamine sulphate, but the clinical importance of these differences was not clear. In two studies of glucosamine sulphate, the need for knee arthroplasty was reduced from 14.5% to 6.3% at 8 years' follow-up. For other preparations of glucosamine, chondroitin and combination therapy, there was less evidence to support a clinical effect. Cost-effectiveness modelling was restricted to glucosamine sulphate. Over a lifetime horizon the incremental cost per quality-adjusted life-year (QALY) gain for adding glucosamine sulphate to current care was estimated to be 21,335 pounds. Deterministic sensitivity analysis suggested that the cost-effectiveness of glucosamine sulphate therapy was particularly dependent on the magnitude of the quality of life (QoL) gain, the change in knee arthroplasty probability with therapy and the discount rate. At a cost per QALY gained threshold of 20,000 pounds, the likelihood that glucosamine sulphate is more cost-effective than current care is 0.43, while at a threshold of 30,000 pounds, the probability rises to 0.73. Probabilistic sensitivity analysis showed that estimates were imprecise and subject to a degree of decision uncertainty. Value of information analysis demonstrated the need for further research. Several biologically plausible mechanisms of action for glucosamine sulphate and chondroitin were proposed. CONCLUSIONS: There was evidence that glucosamine sulphate shows some clinical effectiveness in the treatment of OA of the knee. No trial data came from the UK and caution should be exercised in generalising the findings to the UK health-care setting. Cost effectiveness was not conclusively demonstrated. There was evidence to support the potential clinical impact of glucosamine sulphate. The value of information analysis identified three research priorities: QoL, structural outcomes and knee arthroplasty. The biological mechanism of glucosamine sulphate and chondroitin remains uncertain and, in particular, the proposal that the active substance may be sulphate should be explored further. PMID- 19903417 TI - Do COX-2 inhibitors reduce metachronous cancer development? PMID- 19903418 TI - Gastric acid hypersecretory states: recent insights and advances. AB - Gastric acid hypersecretory states are characterized by basal hypersecretion of gastric acid and historically include disorders associated with hypergastrinemia, hyperhistaminemia, and those of unknown etiology. Although gastric acid secretion is infrequently measured, it is important to recognize the role of gastric hypersecretion in the symptoms of these disorders because they share several features of pathogenesis and treatment. In this article, recent important articles reporting insights into their diagnosis, pathogenesis, and treatment are reviewed. Particular attention is paid to Zollinger-Ellison syndrome, because it has the most extreme acid hypersecretion of this group of disorders and because numerous recent articles deal with various aspects of the diagnosis, molecular pathogenesis, and treatment of the gastrinoma itself or the acid hypersecretion. Two new hypersecretory disorders are reviewed: rebound acid hypersecretion after the use of proton pump inhibitors and acid hypersecretion with cysteamine treatment in children with cystinosis. PMID- 19903419 TI - Gut sensing mechanisms. AB - The mechanisms by which the gut senses and responds to nutrients involve the interplay of multiple complex pathways. In addition to regulating digestion and absorption, the pathways stimulated by molecules in the gut lumen mediate gastric motility, food intake, and satiety. Furthermore, protective mechanisms are activated as necessary to prevent injury, promote healing, and limit intake and absorption of potentially toxic substances. This review provides an update on the current knowledge and recent findings related to gastric sensing of nutrients, highlighting recent research and future endeavors in the field. PMID- 19903420 TI - Regulation of food intake: the gastric X/A-like endocrine cell in the spotlight. AB - Nutritional status influences hormone secretion from specialized enteroendocrine cells within the gut mucosa. These hormones regulate food intake by mediating information to central neurocircuitries in the brainstem and forebrain (eg, hypothalamic nuclei). Intestinal enteroendocrine cells were believed to be the main source of gut peptides regulating food intake. However, recent evidence highlights a specific endocrine cell within the oxyntic glands of the stomach as an important player in appetite control. Acylated ghrelin is the only known orexigenic hormone peripherally produced in gastric X/A-like cells and centrally acting to stimulate food intake. Recent advances led to the assumption that des acylated ghrelin, coreleased with acylated ghrelin, is also involved in regulating food intake. This, and the novel observation that nesfatin-1, which inhibits food intake, is expressed in ghrelin-producing cells of the stomach, supports an important role for gastric X/A-like cells in regulating food intake. Another peptide, obestatin, was initially described as a ghrelin gene product inhibiting food intake, but subsequent studies produced controversial data and its action as an anorexic factor is doubtful. Importantly, synergistic interactions between ghrelin and intestinal peptides seem to orchestrate food intake and body weight regulation, which may have implications for understanding mechanisms leading to the treatment of obesity. PMID- 19903421 TI - Gastric infection by Helicobacter pylori. AB - Helicobacter pylori infection causes chronic active gastritis, ulcer disease, and gastric cancer. Current eradication regimens use a proton pump inhibitor (PPI) and two antibiotics. Triple therapy now has a success rate less than 80%, below the cutoff for efficacious eradication. Antibiotic resistance, inconsistent acid control by PPIs, and poor patient compliance contribute to the failure rate. H. pylori is a neutralophile that has developed special acid acclimation mechanisms to colonize its acidic gastric niche. Identifying the components of these mechanisms will provide novel bactericidal drug targets. Alternatively, better 24 hour acid control would increase the efficacy of antibiotics, leading to dual therapy with improved PPIs and amoxicillin. Studies of acid acclimation by H. pylori have identified several potential eradication targets including UreI, alpha-carbonic anhydrase, and a two-component system. Continuing improvement of PPIs has led to the development of at least three candidate drugs with improved 24-hour acid control. PMID- 19903422 TI - Gastric acid inhibition in the treatment of peptic ulcer hemorrhage. AB - Upper gastrointestinal bleeding from peptic ulcer disease is a common clinical event, resulting in considerable patient morbidity and significant health care costs. Inhibiting gastric acid secretion is a key component in improving clinical outcomes, including reducing rebleeding, transfusion requirements, and surgery. Raising intragastric pH promotes clot stability and reduces the influences of gastric acid and pepsin. Patients with high-risk stigmata for ulcer bleeding (arterial bleeding, nonbleeding visible vessels, and adherent clots) benefit significantly from and should receive high-dose intravenous proton pump inhibitors (PPIs) after successful endoscopic hemostasis. For patients with low risk stigmata (flat spots or clean ulcer base), oral PPI therapy alone is sufficient. For oozing bleeding (an intermediate risk finding), successful endoscopic hemostasis and oral PPI are recommended. Using intravenous PPIs before endoscopy appears to reduce the frequency of finding high-risk stigmata on later endoscopy, but has not been shown to improve clinical outcomes. High-dose oral PPIs may be as effective as intravenous infusion in achieving positive clinical outcomes, but this has not been documented by randomized studies and its cost effectiveness is unclear. PMID- 19903423 TI - Insights into IBD Pathogenesis. AB - Inflammatory bowel disease (IBD), encompassing Crohn's disease (CD) and ulcerative colitis (UC), is a chronic inflammatory disorder caused by dysregulated immune responses in a genetically predisposed individual. Recent accumulating data, including genome-wide association studies, have identified more than 50 distinct genetic loci that confer susceptibility. We highlight the role of microbial-host interaction, particularly with respect to the overlap of common genetic and pathophysiologic mechanisms of CD and UC, interleukin-22 producing natural killer cells, autophagy, and TL1A, a member of the tumor necrosis factor (TNF) family, in gut homeostasis and IBD pathogenesis. This article focuses on the recent advances in understanding of IBD from the past year, including advances in genetics and immunobiology. PMID- 19903424 TI - Natural history and complications of IBD. AB - Inflammatory bowel diseases (IBD), mainly ulcerative colitis and Crohn's disease, are chronic, heterogenic, lifelong illnesses with young age of onset and a great potential for disability. The natural history of these diseases is influenced by multiple factors of environmental and genetic origin. Multidisciplinary research has increased our knowledge of the mechanisms involved during the development and outcome of the diseases, including disease complications. Immunomodulatory treatment has demonstrated greatly improved efficacy in moderate to severe disease activity. The long-term effect on the natural course of disease and sustained reduced burden on society over many years require study. This article summarizes recent knowledge on factors influencing the natural history of IBD, including the impact of treatment. Increased understanding of disease mechanisms is needed as a basis for new treatment strategies in the future. PMID- 19903425 TI - Diagnostic advances in inflammatory bowel disease (imaging and laboratory). AB - Autoimmune and antimicrobial antibodies currently play only an adjunctive role in the diagnosis of inflammatory bowel disease (IBD). Their sensitivity and specificity are not high enough to be relied upon alone to secure a diagnosis; however, their most promising role seems to be in identifying Crohn's disease patients at a higher risk of progression to intestinal complications. Serum C reactive protein (CRP) correlates well with other measures of biologic activity but not as well with clinical activity. CRP can help predict IBD relapses, and in patients with severely active ulcerative colitis may indicate which patients are most likely to progress to colectomy. Similarly, fecal lactoferrin and calprotectin are reasonably accurate and noninvasive measures of disease activity, can predict relapse, and identify a high-risk group among acute severe colitis patients. Capsule endoscopy is a highly sensitive tool that can be used in patients with suspected Crohn's disease with a negative traditional workup, but lower specificity and the risk of capsule retention preclude first-line use. CT enterography can delineate the extent and severity of bowel inflammation and detect extraluminal findings. Magnetic resonance enterography is a radiation-free cross-sectional imaging alternative that is comparable to CT enterography in diagnostic accuracy. PMID- 19903426 TI - Optimizing conventional therapies for inflammatory bowel disease. AB - Conventional therapies remain the mainstay of treatment for most patients with inflammatory bowel disease (IBD), with only a minority of patients requiring biologic therapies. Recently, attention has focused on optimizing dosing strategies for biologic agents; however, of equal importance are recent advances in the optimization of conventional IBD therapies. Newer aminosalicylate formulations demonstrate similar efficacy with a reduced pill burden and less frequent dosing, while new corticosteroid preparations may retain efficacy with a significantly improved safety profile. The limited indications for antibiotics and probiotics have been further refined by recent data, although uncertainties remain. Advances in the understanding of thiopurine metabolism continue to improve dose optimization and the potential for deliberate therapeutic manipulation with adjunctive therapies. An improved knowledge of intracellular methotrexate metabolism may translate to similar opportunities in the future. This article discusses recent advances relevant to clinical practice today. PMID- 19903427 TI - Optimizing biologic therapies for inflammatory bowel disease (ulcerative colitis and Crohn's disease). AB - The introduction of biologic agents and particularly of anti-tumor necrosis factor antibodies dramatically changed the therapeutic algorithm in patients with inflammatory bowel diseases. Although the efficacy of these agents has been demonstrated clearly, optimal treatment strategies are debated. Recent trials advocate the introduction of biologic agents at an early stage to prevent debilitating complications. However, significant adverse events have led to careful selection of patients who will benefit most from long-term treatment with biologic agents. Once on biologic therapy, scheduled maintenance therapy is recommended to minimize the risk of loss of response. Nevertheless, treatment adaptation is frequently necessary in patients who lose response. Interventions encompass strategies to increase drug exposure by increasing the dose or decreasing the dosing interval, or by changing to another biologic agent. Finally, it remains unclear if and when a biologic agent can be stopped in patients with long-standing remission. PMID- 19903428 TI - Managing the risks of IBD therapy. AB - Successful management of the patient with inflammatory bowel disease (IBD) involves not only the induction and maintenance of remission, but also the optimization of the benefit-to-risk equation to achieve the greatest gain in quality of life. These risks range from intolerance to prescribed medications to potentially life-threatening sequelae (eg, sepsis) of immune suppression. A proper awareness of risk on the part of the physician and education of the patient can lead to early detection and institution of an appropriate management plan, including risk management and, optimally, primary prevention (eg, prophylactic vaccination). One should take the opportunity regularly to reassess the utility and efficacy of existing therapy, with the provision of ineffective therapies mandating urgent review. Overall, optimal management of the patient with IBD requires open dialogue between clinician and patient so that both are cognizant of the goals, benefits, and potential risks of therapy. PMID- 19903429 TI - Satisfying patient expectations with soft-tissue augmentation. AB - The popularity of cosmetic procedures for rejuvenating the face has undergone enormous growth over the past few years, such that at least one in four cosmetic procedures involves the use of soft-tissue augmentation. Of note is the trend away from surgical interventions and toward noninvasive cosmetic procedures, which now account for the majority of all cosmetic procedures performed in the United States. Adult patients of all ages are selecting soft-tissue augmentation, either as a precursor to or a substitute for surgery; there is a trend toward the use of injectable devices in younger patients (aged 35 - 50 years). Patients in different age groups have diverse treatment needs ranging from the correction of fine lines and wrinkles in younger patients to volume restoration in older patients. Thus, the treatment needs will dictate the treatment approach, particularly with injectable filler treatments. The aesthetic physician needs to help the patient understand and select the most appropriate rejuvenating treatment based on a variety of factors, specifically, patient age, motivating factors, timing, cosmetic area to be addressed, and desired outcome. This article suggests a series of steps to help determine the most appropriate approach for volume restoration with injectable devices for satisfying patient treatment expectations. PMID- 19903430 TI - Nail disease in pemphigus vulgaris. AB - Patients affected by pemphigus vulgaris will occasionally present with associated and characteristic nail changes. This manuscript was prepared as a review of this unique presentation of pemphigus. Articles describing pemphigus vulgaris of the nail were compiled and reviewed and pertinent information was extracted to provide a concise analysis. The literature on this topic is still maturing. Publications to date suggest the incidence of nail and periungal involvement may correlate with the severity of mucocutaneous symptoms and the duration of the disease. The most common nail manifestations reported were acute or chronic paronychia and onychomadesis, and involvement of the fingernails. The nail changes may precede or develop concurrently with the mucocutaneous presentation. The diagnosis can be made by histologic identification and/or by the presence of IgG and C3 with direct immunoflourescence testing. Systemic treatment is required and highly effective; it includes a tailored combination of anti-inflammatory medications, immunosuppressive agents, and/or immunomodulatory treatments. This review aims to provide a better understanding of pemphigus vulgaris nail changes and guidance for clinical treatment. Unknown etiologies, predisposing factors, discrepancies evident in the literature, and limited research warrant further investigation of pemphigus vulgaris of the nail. PMID- 19903431 TI - [Pyoderma gangrenosum--clinical, laboratory and therapeutic approaches. Review of 28 cases]. AB - Pyoderma gangrenosum is an uncommon neutrophilic dermatosis, idiopatic in 25-50 percent of cases. We present a retrospective study of 28 patients, admitted to Hospital de Sao Joao, EPE, between January of 1990 and June of 2008. Pyoderma gangrenosum was observed more frequently in middle age adults, in the ulcerated form, and predominantly localized in the lower limbs. Pediatric presentation was very rare. In 50 percent of these cases a systemic disease was present and the most frequent was Crohn disease (14% of patients). PMID- 19903432 TI - Advice for medical students interested in dermatology: perspectives from fourth year students who matched. AB - We present perspectives from four fourth year medical students who matched into dermatology that highlight the factors they believed helped them most. The purpose is to offer advice to medical students interested in dermatology. We divide the paper into four areas of discussion: academics, extracurricular activities, research, and mentorship. All four factors are crucial for a strong dermatology application. We believe the paper provides valuable suggestions and guidance to students considering a career in dermatology. PMID- 19903433 TI - A case of bullous disease limited to the skin illustrates the spectrum of neoplasia-induced autoimmunity. AB - Although the initial report of paraneoplastic pemphigus described individuals with mucocutaneous blistering disease, subsequent reports identified patients with lichen planus or graft versus host disease-like changes. We describe a patient with fatal autoimmune blistering disease with absence of mucous membrane lesions. The pattern of complement indirect immunofluoresence helped identify the prognosis prospectively. This case illustrates yet another presentation of the neoplasia-induced autoimmunity. PMID- 19903434 TI - A case of primary cutaneous B-cell lymphoma of the knee. AB - We report the case of a gentleman with a primary cutaneous B cell lymphoma of the leg who underwent an amputation and later died. This is an uncommon type of cutaneous lymphoma with poor prognosis and the case demonstrates how aggressive the tumor can become. PMID- 19903435 TI - Pseudoxanthoma-elasticum-like syndrome and thalassemia: an update. AB - Pseudoxanthoma Elasticum (PXE) is an autosomal recessive, multisystem disorder affecting connective tissues. We describe three cases of the acquired PXE-like syndrome that often occurs in association with hemolytic anemias, in particular the hemoglobinopathies, and review the literature on the subject. The pathogenesis of the acquired PXE-like lesions is not yet completely understood. None of the mutations observed in the inherited form has been detected in the syndrome accompanying thalassemia. The cardiovascular complications could be life threatening. Therefore, an close surveillance of these patients is mandatory. PMID- 19903436 TI - Mucoceles not - Oral cysticercosis and minor salivary gland adenocarcinoma: two case reports. AB - A mucocele is one of the more commonly encountered disorders of the oral mucosa. It develops when a minor salivary duct is injured or blocked, resulting i n the escape of mucus into the adjacent submucosal connective tissue. We present two cases that were clinically diagnosed as mucoceles that were removed because of patient anxiety. However, histological examination revealed dissimilar and medically significant pathologies. Oral cysticercosis and salivary gland adenocarcinoma are rare diseases of the oral mucosa. It is important to consider these and other uncommon diagnoses when evaluating an oral nodule presumed to be a mucocele. We will discuss the natural history and pathogenesis of these disease processes. PMID- 19903437 TI - Keratoacanthoma in a tattoo. AB - Several malignant lesions have been reported in association with tattoos, including basal cell carcinomas, squamous cell carcinomas and melanoma. We report a keratoacanthoma arising in a recent tattoo. A 60-year-old woman presented with a tumor on her right leg, over an area of red pigment in a professional tattoo. Histological analysis confirmed the clinical diagnosis of keratoacanthoma. Although the association between malignancy and tattoos is very uncommon dermatologists and dermatologic surgeons should be attentive to this possibility. PMID- 19903439 TI - The diagnosis. PMID- 19903438 TI - A remarkable case of cutaneous metastatic breast carcinoma. AB - We describe a 50-year-old woman with a 5-month history of multiple asymptomatic papulonodular lesions on the left chest area. Biopsy was consistent with cutaneous metastases from a ductal breast carcinoma. No distant metastatic lesions were detected. The patient was referred to the Gynecologic Oncology Department. Treatment included chemotherapy, radiotherapy and surgery. At present the patient is well with no signs of recurrence. This case reports a clinically remarkable cutaneous metastatic breast carcinoma. PMID- 19903440 TI - Autologous "foreign body" as a sequel of improper cutting of an ingrowing toe nail? AB - We report on a 63-year-old male who suffered from an ingrown toenail affecting the left first digit for several years. Medical history revealed that repeated vigorous nail plate trimming by the patient relieve the red, painful swelling of his great toe. Furthermore, Emmert onychoplasty as well as non-invasive procedures did not achieve improvement. A thorough surgical exploration of the affected area detected a nail spicule in the deeper paronychium. This was likely caused by improper cutting of the nail plate. Removal of the fragment in combination with partial nail plate excision, followed by phenol cauterization of the matrix resulted in full recovery. PMID- 19903441 TI - Saphenous vein harvesting site dermatoses in eastern India. AB - Coronary artery bypass grafting (CABG) has become a common surgical procedure. During this procedure, the greater saphenous vein is most often used as the conduit and the side effects of the leg vein harvesting include a variety of cutaneous changes. We report the cutaneous effects at the saphenous vein harvesting sites in 21 patients who underwent CABG. PMID- 19903442 TI - Drug-induced pruritic micropapular eruption: anastrozole, a commonly used aromatase inhibitor. AB - Anastrozole, a selective nonsteroidal aromatase inhibitor is widely used as an adjuvant therapy for postmenopausal women with early hormone-sensitive breast cancer. There are few reports on cutaneous side effects of anastrozole. It may induce subacute cutaneous lupus erythematosus, erythema nodosum, cutaneous vasculitis, and nondescript skin eruptions. A 68-year-old woman was prescribed anastrozole after surgical removal of her breast cancer and adjuvant radiation therapy. Two months later she experienced a generalized pruritic micropapular eruption. History, clinical presentation, histology and inadvertent re-exposure to the drug confirmed that anastrozole triggered the exanthem. Pruritic micropapular eruption is a typical pattern for a drug hypersensitivity reaction. Anastrozole should be added to the list of medications able to induce not only non-specific eruptions but the type of exanthem typically triggered by drugs. PMID- 19903443 TI - Adalimumab in the management of palmoplantar psoriasis. AB - Palmoplantar pustular psoriasis (PPP) is an uncommon form of chronic psoriasis. Characterized by sterile, intraepidermal pustules located on the palms and soles, it is highly resistant to treatment. Our patient presented with palmar inflammation and throbbing joint pain in his hands, as well as erythematous, pustular, and micaceous scaling skin on his right foot, legs, elbows, and hands. Approximately 4 percent of his body surface area was involved and he was diagnosed with PPP after skin biopsy. After conventional therapy failed, the patient underwent treatment with adalimumab and the majority of his symptoms resolved after 16 weeks of therapy. Adalimumab may be effective for the treatment of PPP. Adalimumab, a fully human immunoglobulin G1 monoclonal antibody that binds to tumor necrosis factor, has been approved for the treatment of moderate to severe psoriasis, in the United States, Europe, and elsewhere. PMID- 19903444 TI - Quantitation of folates and their catabolites in blood plasma, erythrocytes, and urine by stable isotope dilution assays. AB - New stable isotope dilution assays were developed for the simultaneous quantitation of the folates 5-methyltetrahydrofolic acid, 5-formyltetrahydrofolic acid, tetrahydrofolic acid, 10-formylfolic acid, and folic acid as well as for their catabolites para-aminobenzoylglutamate (pABG) and acetyl-para aminobenzoylglutamate (ApABG) in clinical samples. The methods were based on cleanup by strong anion exchange followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) detection. Deuterated analogues of the folates and [(13)C(5)]-labeled isotopologues of the catabolites were applied as the internal standards in stable isotope dilution assays. Extraction in 4 morpholineethanesulfonic acid (MES) buffer at pH 5.0 ensured the optimum stability of folates and, in combination with solid-phase extraction (SPE) based on strong anion exchange, resulted in higher recoveries compared with other combinations of extraction buffers and SPE. The method was sensitive enough to detect pABG in plasma generally and unmetabolized folic acid in the plasma of a volunteer after oral dosage of an aqueous folic acid solution. The sum of folate catabolites increased by a factor of 2 in the urine of the latter volunteer, compared with that resulting when only water was dosed. PMID- 19903445 TI - Selection of thiol- and disulfide-containing proteins of Escherichia coli on activated thiol-Sepharose. AB - Activated thiol-Sepharose (ATS) facilitates selection of thiol-containing proteins. In control- and menadione-treated Escherichia coli, batch selection performed under denaturing conditions revealed distinct two-dimensional electrophoresis (2DE) patterns. Using shotgun proteomics, 183 thiol-containing proteins were identified in control ATS-selected extracts and 126 were identified in menadione-treated E. coli, with 85 proteins being common to both. More than 90% of identified proteins contained one or more cysteines. Blocking with N-ethyl maleimide followed by reduction facilitated ATS-based selection of disulfide containing proteins. In total, 62 proteins were unique to control cells and 164 were identified in menadione-treated E. coli cells, with 29 proteins being common to both. Proteins from menadione-treated cells were excised from 2DE gels, digested with trypsin, and identified by peptide mass fingerprinting. This revealed 19 unique proteins, 14 of which were identified by shotgun proteomics. Outer membrane proteins A, C, W, and X and 30S ribosomal protein S1 were found in 2DE but not by shotgun proteomics. Foldases, ribosomal proteins, aminoacyl transfer RNA (tRNA) synthetases, and metabolic and antioxidant enzymes were prominent among identified proteins, and many had previously been found to respond to, and be targets for, oxidative stress in E. coli. ATS provides a convenient and rapid way to select thiol-containing proteins. PMID- 19903446 TI - Facile construction of fluorescent peptide microarrays: One-step fluorescent derivatization of sub-microscale peptide aldehydes for selective terminal immobilization. AB - In this note, we demonstrate the utility of bifunctional fluorescent linkers to facilitate the construction of peptide microarrays with either an N- or a C terminal alkylamine for directionally preferred peptide immobilization. Significantly, these small tags facilitate high-performance liquid chromatography (HPLC) profiling while limiting interference with antigen-antibody interactions after peptide immobilization. In a model peptide-antibody binding assay, a sequence-dependent orientation effect of antibody binding to a series of peptide ligands was demonstrated. This approach provides a strategy that can be applied to a variety of peptide microarray-based detection systems. PMID- 19903447 TI - Template-blocking PCR: an advanced PCR technique for genome walking. AB - This article describes the development of an improved method for the isolation of genomic fragments adjacent to a known DNA sequence based on a cassette ligation mediated polymerase chain reaction (PCR) technique. To reduce the nonspecific amplification of PCR-based genome walking, the 3' ends of the restriction enzyme digested genomic DNA fragments were blocked with dideoxynucleoside triphosphate (ddNTP) and ligated with properly designed cassettes. The modified genomic DNA fragments flanked with cassettes were used as a template for the amplification of a target gene with a gene-specific primer (GSP) and a cassette primer (CP). The ddNTP blocking of the genomic DNA ends significantly reduced the nonspecific amplification and resulted in a simple and rapid walking along the genome. The efficiency of the template-blocking PCR method was confirmed by a carefully designed control experiment. The method was successfully applied for the cloning of the PGK1 promoter from Pichia ciferrii and two novel cellulase genes from Penicillium sp. PMID- 19903448 TI - Bacteria capture, lysate clearance, and plasmid DNA extraction using pH-sensitive multifunctional magnetic nanoparticles. AB - A multifunctional magnetic nanoparticle (MNP)-assisted bioseparation method was developed to isolate plasmid DNA (pDNA) from Escherichia coli culture. Using the pH-sensitive carboxyl-modified magnetic nanoparticles, both cell capture and the subsequent removal of genomic DNA/protein complex after lysis can be achieved simply by magnetic separation. Furthermore, the yield and purity of pDNA extracted by MNPs are comparable to those obtained using organic solvents or commercial kits. This time- and cost-effective protocol does not require centrifugation or precipitation steps and has the potential for automated DNA extraction, especially within miniaturized lab chip applications. PMID- 19903449 TI - 7-Benzyloxyresorufin-O-dealkylase activity as a marker for measuring cytochrome P450 CYP3A induction in mouse liver. AB - The cytochrome P450 subfamily CYP3A belongs to the most important detoxification enzymes. Because the main CYP3A isoforms are not polymorphic and therefore detract themselves from genetic screening as a potent prediction marker for drug metabolism or induction effects, effective in vitro testing of a putative drug CYP3A interaction is indicated. We used mouse liver microsomes treated with the model drug phenytoin to set up an effective and reliable in vitro test system. A metabolic assay analyzing 7-alkoxyresorufin-O-dealkylation showed specific CYP3A dependent 7-benzyloxyresorufin oxidation (BROD). This was confirmed by testing other alkoxyresorufins (7-ethoxy-, 7-methoxy-, and 7-pentoxyresorufin) in mice and correlation of the data with testosterone 6beta-hydroxylation and a plethora of isoform-specific chemical inhibitors (orphenadrine, chloramphenicol, nifedipine, ketoconazole, and sulfaphenazole). Isoform-specific expression and induction of CYP3A11 in mouse liver was tested by RNase protection assay, reverse transcription polymerase chain reaction (RT-PCR), and immunoblot. With the BROD assay, we could clearly dissect CYP3A11 from other P450s induced by phenytoin like CYP2C29, CYP2B9, CYP1A1, and CYP4A. We conclude that the BROD assay is a specific tool to assign CYP3A induction by drugs or other chemicals, at least in a mouse model system. PMID- 19903450 TI - High-resolution melting analysis for genotyping of the myotonic dystrophy type 1 associated Alu insertion/deletion polymorphism. AB - Since its introduction, high-resolution melting (HRM) analysis has been used for genotyping of various types of sequence alterations. In this study, we report the use of HRM for genotyping of the 1-kb insertion/deletion polymorphism, involving a problematic region of five consecutive Alu elements, that is associated with myotonic dystrophy type 1. We combined a three-primer polymerase chain reaction (PCR) amplification approach with HRM using two primer sets. Analyses based on curve shapes are sensitive enough to differentiate between genotypes with both primer sets. In addition, the newly designed insertion-specific primer from the second primer set equalizes the allele-specific amplicon lengths, thereby reducing the possibility of preferential amplification of shorter fragments. PMID- 19903451 TI - Divalent cations induce a compaction of intrinsically disordered myelin basic protein. AB - Central nervous system myelin is a dynamic entity arising from membrane processes extended from oligodendrocytes, which form a tightly-wrapped multilamellar structure around neurons. In mature myelin, the predominant splice isoform of classic MBP is 18.5kDa. In solution, MBP is an extended, intrinsically disordered protein with a large effective protein surface for myriad interactions, and possesses transient and/or induced ordered secondary structure elements for molecular association or recognition. Here, we show by nanopore analysis that the divalent cations copper and zinc induce a compaction of the extended protein in vitro, suggestive of a tertiary conformation that may reflect its arrangement in myelin. PMID- 19903452 TI - DGDA, a local sequence of the kringle 2 domain, is a functional motif of the tissue-type plasminogen activator's antiangiogenic kringle domain. AB - Antiangiogenic activity can be elicited by the kringle domains 1 and 2 of tissue type plasminogen activator (TK1-2), or the kringle 2 domain alone. In a previous report, we showed that the anti-migratory effect of TK1-2 is mediated in part by its interference with integrin alpha2beta1. Since integrin alpha2beta1 interacts with collagen type I through the DGEA (Asp-Gly-Glu-Ala) amino acid sequence, and a similar sequence, DGDA (Asp-Gly-Asp-Ala), exists in the kringle 2 domain, we investigated whether the DGDA sequence has a role in antiangiogenic activity of TK1-2. In an adhesion assay, the DGDA peptide inhibited adhesion of human umbilical vein endothelial cells (HUVECs) to immobilized TK1-2. Pretreatment of the DGDA peptide also blocked anti-migratory activity of TK1-2. When the DGDA peptide alone was tested for antiangiogenic activity, it effectively inhibited VEGF-induced migration of HUVECs and tube formation on Matrigel. In addition, the DGDA peptide decreased differentiation of endothelial progenitor cells on collagen type I matrix. These data suggest that the DGDA sequence presents a functional epitope of TK1-2 and that it can be used as a potential novel antiangiogenic peptide. PMID- 19903453 TI - Functional and structural characterization of the talin F0F1 domain. AB - The globular head domain of talin, a large multi-domain cytoplasmic protein, is required for inside-out activation of the integrins, a family of heterodimeric transmembrane cell adhesion molecules. Talin head contains a FERM domain that is composed of F1, F2, and F3 subdomains. A F0 subdomain is located N-terminus to F1. The F3 contains a canonical phosphotyrosine binding (PTB) fold that directly interacts with the membrane proximal NPxY/F motif in the integrin beta cytoplasmic tail. This interaction is stabilized by the F2 that interacts with the lipid head-groups of the plasma membrane. In comparison to F2 and F3, the properties of the F0F1 remains poorly characterized. Here, we showed that F0F1 is essential for talin-induced activation of integrin alphaLbeta2 (LFA-1). F0F1 has a high content of beta-sheet secondary structure, and it tends to homodimerize that may provide stability against proteolysis and chaotrope induced unfolding. PMID- 19903454 TI - Letter to the editor on "Threonine 53 in alpha-synuclein is conserved in long living non-primate animals". PMID- 19903455 TI - How does oxygen rise drive evolution? Clues from oxygen-dependent biosynthesis of nuclear receptor ligands. AB - It is well known that oxygen rise greatly facilitated biological evolution. However, the underlying mechanisms remain elusive. Recently, Raymond and Segre revealed that molecular oxygen allows 1000 more metabolic reactions than can occur in anoxic conditions. From the novel metabolites produced in aerobic metabolism, we serendipitously found that some of the metabolites are signaling molecules that target nuclear receptors. Since nuclear signaling systems are indispensable to superior organisms, we speculated that aerobic metabolism may facilitate biological evolution through promoting the establishment of nuclear signaling systems. This hypothesis is validated by the observation that most (97.5%) nuclear receptor ligands are produced by aerobic metabolism, which is further explained in terms of the chemical criteria (appropriate volume and rather high hydrophobicity) of nuclear receptor ligands that aerobic metabolites are more ready than anaerobic counterparts to satisfy these criteria. PMID- 19903456 TI - AMP-activated protein kinase is activated in Parkinson's disease models mediated by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. AB - The selective loss of dopaminergic neurons in the substantia nigra pars compacta is a feature of Parkinson's disease (PD). 1-Methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP)-induced neurotoxicity is the most common experimental model used to investigate the pathogenesis of PD. Administration of MPTP in mice produces neuropathological defects as observed in PD and 1-methyl-4-pyridinium (MPP(+)) induces cell death when neuronal cell cultures are used. AMP-activated protein kinase (AMPK) is a key regulator of energy homeostasis. In the present study, we demonstrated that AMPK is activated by MPTP in mice and MPP(+) in SH SY5Y cells. The inhibition of AMPK by compound C resulted in an increase in MPP(+)-induced cell death. We further showed that overexpression of AMPK increased cell viability after exposure to MPP(+) in SH-SY5Y cells. Based on these results, we suggest that activation of AMPK might prevent neuronal cell death and play a role as a survival factor in PD. PMID- 19903457 TI - Engineered drug-resistant immunocompetent cells enhance tumor cell killing during a chemotherapy challenge. AB - Establishment of immunocompetent cell mediated anti-tumor immunity is often mitigated by the myelosuppressive effects during administration of chemotherapy. We hypothesized that protecting these immune cells from drug induced toxicities may allow for the combined administration of immunotherapy and chemotherapy. Using a SIV-based lentiviral gene transfer system we delivered the drug-resistant variant P140KMGMT into the immunocompetent cell lines NK-92 and TALL-104, and the myelogenous leukemia cell line, K562, which is a target for both NK-92 and TALL 104 cells. Genetically engineered immunocompetent cells developed significant resistance to temozolomide compared to non-modified cells, and genetic modification of these cells did not affect their ability to kill K562 cells. We then evaluated the effectiveness of drug-resistant immunocompetent cell mediated killing of tumor cells in the presence and absence of chemotherapy. During a chemotherapy challenge the cytotoxic activity of non-modified immunocompetent cells was dramatically impaired. However, when combined with chemotherapy, genetically-modified immune cells retained their cytotoxic activities and efficiently killed non-modified target cells. These results show that engineering immunocompetent cells to withstand chemotherapy challenges can enhance tumor cell killing when chemotherapy is applied in conjunction with cell-based immunotherapy. PMID- 19903458 TI - p52-Independent nuclear translocation of RelB promotes LPS-induced attachment. AB - The NF-kappaB signaling pathways have a critical role in the development and progression of various cancers. In this study, we demonstrated that the small cell lung cancer cell line (SCLC) H69 expressed a unique NF-kappaB profile as compared to other cancer cell lines. The p105/p50, p100/p52, c-Rel, and RelB protein and mRNA transcripts were absent in H69 cells but these cells expressed RelA/p65. The activation of H69 cells by lipopolysaccharide (LPS) resulted in the induction of RelB and p100 expression. The treatment also induced the nuclear translocation of RelB without the processing of p100 to p52. Furthermore, LPS induced beta1 integrin expression and cellular attachment through an NF-kappaB dependent mechanism. Blocking RelB expression prevented the increase in the expression of beta1 integrin and the attachment of H69. Taken together, the results suggest that RelB was responsible for the LPS-mediated attachment and may play an important role in the progression of some cancers. PMID- 19903459 TI - Perspectives on the formation of an interdisciplinary research team. AB - As research funding becomes more competitive, it will be imperative for researchers to break the mentality of a single laboratory/single research focus and develop an interdisciplinary research team aimed at addressing real world challenges. Members of this team may be at the same institution, may be found regionally, or may be international. However, all must share the same passion for a topic that is bigger than any individual's research focus. Moreover, special consideration should be given to the professional development issues of junior faculty participating in interdisciplinary research teams. While participation may be "humbling" at times, the sheer volume of research progress that may be achieved through interdisciplinary collaboration, even in light of a short supply of grant dollars, is remarkable. PMID- 19903460 TI - Fibroblast growth factor-2 induces osteogenic differentiation through a Runx2 activation in vascular smooth muscle cells. AB - Expression of bone-associated proteins and osteoblastic transcription factor Runx2 in arterial cells has been implicated in the development of vascular calcification. However, the signaling upstream of the Runx2-mediated activation of osteoblastic program in vascular smooth muscle cells (VSMC) is poorly understood. We examined the effects of fibroblast growth factor-2 (FGF-2), an important regulator of bone formation, on osteoblastic differentiation of VSMC. Stimulation of cultured rat aortic SMC (RASMC) with FGF-2 induced the expression of the osteoblastic markers osteopontin (OPN) and osteocalcin. Luciferase assays showed that FGF-2 induced osteocyte-specific element (OSE)-dependent transcription. Downregulation of Runx2 by siRNA repressed the basal and FGF-2 stimulated expression of the OPN gene in RASMC. FGF-2 produced hydrogen peroxide in RASMC, as evaluated by fluorescent probe. Induction of OPN expression by FGF-2 was inhibited not only by PD98059 (MEK1 inhibitor) and PP1 (c-Src inhibitor), but also by an antioxidant, N-acetyl cysteine. Nuclear extracts from FGF-2-treated RASMC exhibited increased DNA-binding of Runx2 to its target sequence. Immunohistochemistry of human coronary atherectomy specimens and calcified aortic tissues showed that expression of FGF receptor-1 and Runx2 was colocalized. In conclusion, these results suggest that FGF-2 plays a role in inducing osteoblastic differentiation of VSMC by activating Runx2 through mitogen activated protein kinase (MAPK)-dependent- and oxidative stress-sensitive signaling pathways. PMID- 19903461 TI - Structural and dynamic changes of the serum response element and the core domain of serum response factor induced by their association. AB - Transcriptional activity of serum response factor (SRF) is dependent on its binding to the CC(A/T)(6)GG box (CArG box) of serum response element (SRE). By Raman spectroscopy, we carried out a comparative analysis, in solution, of the complexes obtained from the association of core-SRF with 20-mer SREs bearing wild type and mutated c-fos CArG boxes. In case of association with the wild type c fos CArG box, the complex does not bring out the expected Raman signature of a stable bending of the targeted SRE but keeps a bend-linear conformer oligonucleotide interconversion. The linear conformer population is larger than that of free oligonucleotide. In the core-SRF moiety of the wild-type complex a large spectral change associated with the CO-groups from Asp and/or Glu residues shows that their ionization states and the strength of their interactions decrease as compared to those of mutated non-specific complexes. Structural constraints evidenced on the free core-SRF are released in the wild-type complex and environmental heterogeneities appear in the vicinity of Tyr residues, due to higher water molecule access. The H-bonding configuration of one Tyr OH-group, in average, changes with a net transfer from H-bond acceptor character to a combined donor and acceptor character. A charge repartition distributed on both core-SRF and targeted SRE stabilizes the specific complex, allowing the two partners to experience a variety of conformations. PMID- 19903462 TI - Senescence-related functional nuclear barrier by down-regulation of nucleo cytoplasmic trafficking gene expression. AB - One of the characteristic natures of senescent cells is the hypo- or irresponsiveness not only to growth factors but also to apoptotic stress. In the present study, we confirmed the inhibition of nuclear translocation of activated p-ERK1/2 and NF-kB p50 in response to growth stimuli or LPS in the senescent human diploid fibroblasts. In order to elucidate the underlying mechanism for the senescence-associated hypo-responsiveness, we carried out the comparison study for gene expression profiles through microarray analysis. In consequence, we observed the vast reduction in expression of nucleo-cytoplasmic trafficking genes in senescent cells, when compared with those in young cells. Expression levels of several nucleoporins, karyopherin alpha, karyopherin beta, Ran, and Ran regulating factors were confirmed to be down-regulated in senescent HDFs by using RT-PCR and Western blot methods. Taken together, these data suggest the operation of certain senescence-associated functional nuclear barriers by down-regulation of the nucleo-cytoplasmic trafficking genes in the senescent cells. PMID- 19903463 TI - Molecular analysis and prognostic impact of the novel apoptotic gene BCL2L12 in gastric cancer. AB - Stomach cancer comprises a malignancy with feeble prognosis. In gastric carcinogenesis, molecular alterations in the apoptosis-related genes have been described. In this study, the expression of BCL2-like-12 (BCL2L12) gene, discovered and cloned by members of our group, was investigated in a statistically significant sample size of cancerous and non-cancerous stomach tissues and gastric cancer cells with quantitative real-time PCR methodology. BCL2L12 transcript was indicated in cancer gastric tissues to range from 29 to 53200 mRNA copies BCL2L12/10(6) mRNA copies GAPDH. Significant associations of BCL2L12 with gastric tumors of the early stages (I/II) (p=0.044) and of intestinal histotype (p=0.034) was substantiated. Both univariate and multivariate analyses disclosed, respectively, BCL2L12 relationship with disease free (p=0.006 and p=0.025) and overall patients' survival (p=0.007 and p=0.022). Our results open new horizons for the possible application of BCL2L12 as a novel prognostic indicator of gastric cancer. PMID- 19903464 TI - Block effect of capsaicin on hERG potassium currents is enhanced by S6 mutation at Y652. AB - The objectives of this study were to investigate the inhibitory action of capsaicin on wild-type (WT) and mutation human ether-a-go-go-related gene (hERG) potassium channel currents (I(hERG)), and to determine whether mutations in the S6 region are significant for the inhibition of I(hERG) by capsaicin. The hERG channel (WT, Y652A and F656A) was expressed in Xenopus oocytes and studied using standard two-microelectrode voltage-clamp techniques. The results show that capsaicin blocks WT hERG in a concentration-dependent manner, with an IC(50) of 17.45microM and a negative shift in the steady-state inactivation curve. Characteristics of blockade were consistent with capsaicin causing components of block in both the closed and open channel states. However, mutating the Y652 residue to Ala enhances the blockade effect of capsaicin with an IC(50) of 4.11microM, whereas mutation of F656A does not significantly alter drug potency. Simultaneously, for Y652A, the steady-state activation parameter is shifted to a more positive value by 5mV and the inactivation parameter is shifted to a more negative value by -29mV in the presence of 25microM capsaicin. In conclusion, capsaicin blocks hERG channels by binding to both the closed and open channel states.Y652 was important as a molecular determinant of blockade. Mutation Y652A enhances the drug block, which may cause some patients to be particularly sensitive to capsaicin clinically. PMID- 19903465 TI - Decreased number of caveolae in endothelial cells impairs the relaxation induced by acetylcholine in hypertensive rat aortas. AB - The present study was designed to investigate the contribution of endothelial cell caveolae to vascular relaxation in aortas from a normotensive (2K) and renal hypertensive (2K-1C) rat. For that purpose, concentration-effect curves to acetylcholine were constructed in 2K and 2K-1C intact endothelium aortic rings, in the absence or in the presence of the caveolae disassembler methyl-beta ciclodextrin. The potency (pD(2)) and the maximum relaxant effect to acetylcholine were greater in 2K than in 2K-1C aortas. Methyl-beta-ciclodextrin reduced the pD(2) in 2K and the maximum relaxant effect in both 2K and 2K-1C. The quantification of the caveolae number by electronic microscopy has shown a larger number of caveolae in 2K than in 2K-1C endothelial cells, which was reduced by methyl-beta-ciclodextrin in both 2K and 2K-1C. The production of NO stimulated with acetylcholine was greater in 2K than in 2K-1C endothelial cells, and this effect was impaired by methyl-beta-ciclodextrin in both 2K and 2K-1C. The cytosolic Ca(2+) concentration ([Ca(2+)]c) was simultaneously measured in endothelial and smooth muscle cells stimulated with acetylcholine by confocal image of aortic slices. Acetylcholine produced a greater [Ca(2+)]c increase in 2K than in 2K-1C endothelial cells, which response was inhibited by methyl-beta ciclodextrin only in 2K cells. In smooth muscle cells the reduction of [Ca(2+)]c was higher in 2K than in 2K-1C. This effect was inhibited by methyl-beta ciclodextrin only in 2K cells. Taken together, our results suggest that the decreased number of caveolae in the endothelial cells from 2K-1C rat aortas is involved in the impaired effect of acetylcholine on [Ca(2+)]c and NO. PMID- 19903466 TI - Lack of beneficial metabolic effects of quercetin in adult spontaneously hypertensive rats. AB - Insulin sensitivity is partly dependent on insulin-mediated nitric oxide (NO) release and antioxidants may decrease insulin resistance by amelioring NO bioavailability. The effects of chronic therapy with the antioxidant quercetin on blood pressure, vascular function and glucose tolerance in male spontaneously hypertensive rats (SHR), a model of genetically hypertension and insulin resistance, were analyzed. Rats were divided into four groups, WKY vehicle, WKY quercetin, SHR vehicle and SHR quercetin. Animals were daily administered by gavage for four weeks: vehicle, quercetin in vehicle (10mg/kg body weight). Blood pressure was followed by tail-cuff plethysmography. Chronic quercetin treatment reduced systolic blood pressure, and significantly reduced left ventricular ( 10%) and renal (-6%) hypertrophy. However, oral glucose tolerance test, homeostatic model assessment of insulin resistance, total cholesterol and triglycerides were unaffected by quercetin in both strains of rats. It also improved the blunted aortic endothelium-dependent relaxation to acetylcholine, without affecting both endothelium-dependent relaxation to insulin and endothelium-independent relaxation to sodium nitroprusside in SHR. In WKY rats, quercetin in vitro and in vivo, impaired the relaxation to insulin. Quercetin reduced both plasma malondialdehyde levels and aortic superoxide production in SHR. Furthermore, quercetin inhibited insulin-stimulated protein kinase B (Akt)- and endothelial NO synthase (eNOS) phosphorylation. In conclusion, quercetin reduced blood pressure, left ventricular and renal hypertrophy and improved NO dependent acetylcholine relaxation. However, and despite its antioxidant effects, quercetin was unable to improve insulin sensitivity possibly through its specific interference with the insulin signalling pathway. PMID- 19903467 TI - Biomechanical and histological outcome of combined raloxifene-estrogen therapy on skeletal and reproductive tissues. AB - Estrogen replacement is a potent therapy for postmenopausal osteoporosis. However, its carcinogenic effects on breasts and the uterus limit its utilization. Raloxifene has estrogen-like effects on bones without the carcinogenic symptoms on breast or uterine tissue. Their individual effects are well characterized, but the results of their interaction remains elusive. In this work, we investigate the consequences of a combined raloxifene/estrogen therapy on bone and uterus with experimental osteoporosis. 40 Wistar rats began treatment 3 months post-ovariectomy. Estrogen and raloxifene were administered 0.03 mg/kg/day and 1.5mg/kg/day separately and together for 5 times per week for 12 weeks. Biomechanical tests and bone mineral density measurements, histology of uterus, and blood markers were analyzed. The co-administration group had higher toughness and ultimate strength than the ovariectomized controls (P<0.01). E+R had better biomechanical properties than the single treatments; yet the differences were not significant. Uterus histology signified high degeneration in the estrogen group. The raloxifene group had less degeneration but higher vascularization. Less immune reaction and vascularization were observed in the group with combined dosage than in those with individual treatments. Hence, the uterus of the combined treatment had fewer side effects than the ones that were individually treated. Mutual antagonization might be possible between raloxifene and estrogen, and that might have caused a decrease in the adverse effects. Overall, combined therapy might be useful to minimize the individual side effects of raloxifene and estrogen on the uterus and still provide bone strength and toughness. PMID- 19903468 TI - Protein kinase C is inhibited by bisphosphonates in prostate cancer PC-3 cells. AB - Bisphosphonates are expected to be effective at preventing tumor metastasis to bone tissue. Since protein kinase C (PKC) plays a crucial role in cancer progression, we examined the effect of bisphosphonates on PKC expression to clarify the mechanism behind the inhibition of the bone metastasis of prostate cancer by bisphosphonates. We found that pamidronate inhibits PKC protein expression and PKC activity in prostate cancer PC-3 cells. PKC protein expression was markedly reduced by treatment with 100 microM of pamidronate. The inhibitory effect of PKC expression by pamidronate was specific for PKCalpha and PKCzeta. Nitrogen-containing bisphosphonates are known to inhibit the mevalonate pathway, but the effect of pamidronate on PKC expression was not due to the inhibition of this pathway. Urokinase-type plasminogen activator (uPA) is one of the critical proteins in tumor metastasis and decreased in bisphosphonate-treated PC-3 cells. We also showed that uPA expression was suppressed by PKC inhibitors (calphostin C and staurosporine) and induced by a PKC activator (PMA) in PC-3 cells, suggesting that the inhibition of uPA by bisphosphonates is involved in PKC inhibition. This is the first finding that bisphosphonates suppress PKC expression in cancer cells. These results strongly suggest that one of the mechanisms behind the inhibitory effect of bisphosphonates on tumor bone metastasis is mediated by PKC inhibition. PMID- 19903469 TI - XIAP-mediated protection of H460 lung cancer cells against cisplatin. AB - Molecular mechanism(s) responsible for drug resistance of non-small cell lung cancer (NSCLC) cells to cisplatin was investigated. Results showed that cisplatin (50muM)-induced cell death (apoptosis) was more significant in CH27 and A549 cell lines than in H460. The high protein levels of X-linked inhibitor-of-apoptosis protein (XIAP) observed in H460 cells appeared to play a key role in the regulation of cisplatin resistance of H460 cells. XIAP can bind to and suppress the activities of caspase 3 in H460 cells and lead to apoptosis inhibition of these cells. Blockade of XIAP activity by Embelin (XIAP inhibitor) or siRNA has increased caspase 3 activities and promoted cisplatin-induced cell death of H460 cells. The results indicate a therapeutic value of Embelin and/or XIAP siRNA in the control of cisplatin-resistant NSCLC cells (H460). PMID- 19903470 TI - A direct relationship between aggressive behavior in the resident/intruder test and cell oxidative status in adult male mice. AB - Disturbances in oxidative metabolism are involved in many acute and chronic diseases, as well as in several other conditions. The objective of the present study was to examine the relationship between the level of intracellular reactive oxygen species in the peripheral blood granulocytes of mice, as evaluated by 2',7'-dichlorofluorescin diacetate (DCFH-DA), a sensor of reactive oxygen species, and the aggressive behavior of these mice, as estimated by the resident/intruder test. Our results showed a significant, linear and positive relationship (P<0.001) between the intracellular redox status of peripheral blood granulocytes and the aggressive behavior levels of adult male mice (correlation coefficients (R(2)) ranging from 0.75 to 0.77). This suggests that the granulocytes of aggressively behaving mice have high levels of oxidative stress. PMID- 19903471 TI - Selective inhibition of MDR1 (ABCB1) by HM30181 increases oral bioavailability and therapeutic efficacy of paclitaxel. AB - Multi-drug resistance 1 (MDR1, ABCB1), also known as P-glycoprotein (P-gp), restricts intestinal uptake of many drugs, and contributes to cellular resistance to cancer chemotherapy. In this study, we examined the pharmacologic characteristics of HM30181, a newly developed MDR1 inhibitor, and tested its capacity to increase the oral bioavailability and efficacy of paclitaxel, an anti cancer drug usually given by intravenous injection. In the ATPase assay using MDR1-enriched vesicles, HM30181 showed the highest potency (IC(50)=0.63nM) among several MDR1 inhibitors, including cycloporin A, XR9576, and GF120918, and effectively blocked transepithelial transport of paclitaxel in MDCK monolayers (IC(50)=35.4nM). The ATPase inhibitory activity of HM30181 was highly selective to MDR1. HM30181 did not inhibit MRP1 (ABCC1), MRP2 (ABCC2), and MRP3 (ABCC3), and partially inhibited BCRP (ABCG2) only at very high concentrations. Importantly, co-administration of HM30181 (10mg/kg) greatly increased oral bioavailability of paclitaxel from 3.4% to 41.3% in rats. Moreover, oral co administration of paclitaxel and HM30181 showed a tumor-inhibitory strength equal or superior to that of intravenous paclitaxel in the xenograft model in nude mice. These results identify HM30181 as a highly selective and potent inhibitor of MDR1, which in combination with paclitaxel, may provide an orally effective anti-tumor regimen. PMID- 19903472 TI - Plant isoprenoid biosynthesis via the MEP pathway: in vivo IPP/DMAPP ratio produced by (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase in tobacco BY 2 cell cultures. AB - Feeding tobacco BY-2 cells with [2-(13)C,4-(2)H]deoxyxylulose revealed from the (13)C labeling that the plastid isoprenoids, synthesized via the MEP pathway, are essentially derived from the labeled precursor. The ca. 15% (2)H retention observed in all isoprene units corresponds to the isopentenyl diphosphate (IPP)/dimethylallyl diphosphate (DMAPP) ratio (85:15) directly produced by the hydroxymethylbutenyl diphosphate reductase, the last enzyme of the MEP pathway. (2)H retention characterizes the isoprene units derived from the DMAPP branch, whereas (2)H loss represents the signature of the IPP branch. Taking into account the enantioselectivity of the reactions catalyzed by the (E)-4-hydroxy-3 methylbut-2-enyl diphosphate reductase, the IPP isomerase and the trans-prenyl transferase, a single biogenetic scheme allows to interpret all labeling patterns observed in bacteria or plants upon incubation with (2)H labeled deoxyxylulose. PMID- 19903473 TI - Pentagalloylglucose down-regulates mast cell surface FcepsilonRI expression in vitro and in vivo. AB - Mast cell activation by immunoglobulin E (IgE)-mediated stimuli is a central event in the pathogenesis of allergic disorders. The present report shows that treatment with pentagalloylglucose (PGG) resulted in a down-regulation of FcepsilonRI surface expression on mucosal-type murine bone marrow-derived mast cells (mBMMCs), which correlated with a reduction in IgE-mediated activation of mBMMCs. Furthermore, PGG prevented development of allergic diarrhea in a food allergy mouse model and suppressed the up-regulated FcepsilonRI surface expression on mast cells derived from the food-allergy mouse colon. These findings on PGG suggest its therapeutic potential for allergic diseases through suppressing the FcepsilonRI surface expression. PMID- 19903474 TI - Distinct genetic code expansion strategies for selenocysteine and pyrrolysine are reflected in different aminoacyl-tRNA formation systems. AB - Selenocysteine and pyrrolysine, known as the 21st and 22nd amino acids, are directly inserted into growing polypeptides during translation. Selenocysteine is synthesized via a tRNA-dependent pathway and decodes UGA (opal) codons. The incorporation of selenocysteine requires the concerted action of specific RNA and protein elements. In contrast, pyrrolysine is ligated directly to tRNA(Pyl) and inserted into proteins in response to UAG (amber) codons without the need for complex re-coding machinery. Here we review the latest updates on the structure and mechanisms of molecules involved in Sec-tRNA(Sec) and Pyl-tRNA(Pyl) formation as well as the distribution of the Pyl-decoding trait. PMID- 19903475 TI - Mechanical requirements for membrane fission: common facts from various examples. AB - Membrane fission is the last step of membrane carrier formation. As fusion, it is a very common process in eukaryotic cells, and participates in the integrity and specificity of organelles. Although many proteins have been isolated to participate in the various membrane fission reactions, we are far from understanding how membrane fission is mechanically triggered. Here we aim at reviewing the well-described examples of dynamin and lipid phase separation, and try to extract the essential requirements for fission. Then, we survey the recent knowledge obtained on other fission reactions, analyzing the similarities and differences with previous examples. PMID- 19903476 TI - A positively charged amino acid at position 129 in nitrilase from Rhodococcus rhodochrous ATCC 33278 is an essential residue for the activity with meta substituted benzonitriles. AB - A positively charged amino acid (Arg, Lys, or His) at position 129 in Rhodococcus rhodochrous ATCC 33278 nitrilase is essential for the activity of aromatic nitriles. The wild-type enzyme containing Arg129 was active only for meta- and para-substituted benzonitriles with a methyl or amino group, but the R129K and R129H mutant enzymes were active only for meta-substituted benzonitriles. The lack of activity of the mutants for para-substituted benzonitriles may be attributable to steric hindrance between the para-substituent and the side chain of Lys or His. PMID- 19903478 TI - Adenosine-5'-phosphosulfate kinase is essential for Arabidopsis viability. AB - In Arabidopsis thaliana, adenosine-5'-phosphosulfate kinase (APK) provides activated sulfate for sulfation of secondary metabolites, including the glucosinolates. We have successfully isolated three of the four possible triple homozygous mutant combinations of this family. The APK1 isoform alone was sufficient to maintain WT levels of growth and development. Analysis of apk1 apk2 apk3 and apk1 apk3 apk4 mutants suggests that APK3 and APK4 are functionally redundant, despite being located in cytosol and plastids, respectively. We were, however, unable to isolate apk1 apk3 apk4 mutants, most probably because the apk1 apk3 apk4 triple mutant combination is pollen lethal. Therefore, we conclude that APS kinase is essential for plant reproduction and viability. PMID- 19903477 TI - Changeability of individual domains of an aminoacyl-tRNA in polymerization by the ribosome. AB - The changeabilities of individual modules of aminoacyl-tRNAs are poorly understood, despite the relevance for evolution, translational accuracy and incorporation of unnatural amino acids (AAs). Here, we dissect the effect of successive changes in four domains of Ala-tRNA(3)(Ala) on translation in a purified system. Incorporating five AAs, not one, was necessary to reveal major effects on yields of peptide products. Omitting tRNA modifications had little affect, but anticodon mutations were very inhibitory. Surprisingly, changing the terminal CCA to CdCA was sometimes inhibitory and non-cognate AAs were sometimes compensatory. Results have implications for translational fidelity and engineering. PMID- 19903479 TI - Systems biology from a yeast omics perspective. AB - Systems biology represents a paradigm shift from the study of individual genes, proteins or other components to that of the analysis of entire pathways, cellular, developmental, or organismal processes. Large scale studies, primarily initiated in Saccharomyces cerevisiae, have allowed the identification and characterization of components on an unprecedented level. Large scale interaction, transcription factor binding and phosphorylation data have enabled the elucidation of global regulatory networks. These studies have helped provide an understanding of cellular pathways and processes at a global and systems level. PMID- 19903481 TI - Involvement of Src tyrosine kinase in Escherichia coli invasion of human brain microvascular endothelial cells. AB - Invasion of brain microvascular endothelial cells is a prerequisite for successful crossing of the blood-brain barrier by Escherichia coli (E. coli), but the underlying mechanism remains unclear. Here we showed activation of Src tyrosine kinase in E. coli K1 invasion of human brain microvascular endothelial cells (HBMEC). E. coli invasion of HBMEC and the E. coli-induced rearrangement of actin filaments were blocked by Src inhibitors. Overexpression of dominant negative Src in HBMEC significantly attenuated E. coli invasion and the concomitant actin filaments rearrangement. Furthermore, E. coli K1-triggered phosphatidylinositol 3-kinase (PI3K) activation in HBMEC was effectively blocked by Src inhibitors and dominant-negative Src. These results demonstrated the involvement of Src and its interaction with PI3K in E. coli K1 invasion of HBMEC. PMID- 19903480 TI - tRNAs: cellular barcodes for amino acids. AB - The role of tRNA in translating the genetic code has received considerable attention over the last 50 years, and we now know in great detail how particular amino acids are specifically selected and brought to the ribosome in response to the corresponding mRNA codon. Over the same period, it has also become increasingly clear that the ribosome is not the only destination to which tRNAs deliver amino acids, with processes ranging from lipid modification to antibiotic biosynthesis all using aminoacyl-tRNAs as substrates. Here we review examples of alternative functions for tRNA beyond translation, which together suggest that the role of tRNA is to deliver amino acids for a variety of processes that includes, but is not limited to, protein synthesis. PMID- 19903482 TI - c-Abl and Src-family kinases cross-talk in regulation of myeloid cell migration. AB - Cytoskeleton dynamics are regulated by Src-family tyrosine kinases (SFKs) and c Abl. We found that the SFK members Hck and c-Fgr regulate tyrosine phosphorylation of c-Abl and c-Abl associates with beta1 integrin-bound Hck or c Fgr in murine macrophages. Studies with selective inhibitors and cells from SFK deficient mice showed that c-Abl and SFK regulate migration and activation of the small GTPases Cdc42 and Rac in macrophages. Additionally, human neutrophil chemotactic activity was reduced by c-Abl inhibitors, and neutrophils from chronic myeloid leukaemia patients displayed an increased chemotactic ability. Hence, Src-family kinase and c-Abl cross-talk in the regulation of myeloid cell migration. PMID- 19903484 TI - A defined serum-free medium useful for monitoring anti-melanoma responses induced by dendritic cell immunotherapy. AB - Animal sera provide a non-defined source of nutrients and growth factors for mammalian cell culture. Animal serum supplementation may also introduce experimental artefacts, including immune responses against foreign serum proteins. This artefact is particularly apparent in tumour immunotherapy experiments using dendritic cells (DC) and melanoma cells cultured in fetal calf serum (FCS)-replete media. FCS culture of both DC and melanoma cells significantly enhanced anti-tumour responses in mice immunized with DC that had not been pulsed with tumour antigen. Although serum-free media (SFM) may be used for short term culture of cells, most SFM do not support long term culture of tumour cell lines. In addition, in vivo propagation and re-isolation of tumour cells from rodents is expensive, time consuming and only low numbers of viable tumour cells can be recovered from solid tumours. We show that a defined SFM medium is ideal for routine culture of B16 for use in prophylactic DC immunizations, negating the need for in vivo propagation of tumours to avoid FCS effects in tumour implantation experiments. PMID- 19903483 TI - The pedestrian watchmaker: genetic clocks from engineered oscillators. AB - The crucial role of time-keeping has required organisms to develop sophisticated regulatory networks to ensure the reliable propagation of periodic behavior. These biological clocks have long been a focus of research; however, a clear understanding of how they maintain oscillations in the face of unpredictable environments and the inherent noise of biological systems remains elusive. Here, we review the current understanding of circadian oscillations using Drosophila melanogaster as a typical example and discuss the utility of an alternative synthetic biology approach to studying these highly intricate systems. PMID- 19903485 TI - Identification and characterization of HLA-A*0301 epitopes in HIV-1 gag proteins using a novel approach. AB - Identification of CTL epitopes correlated to immune protection is important for the development of vaccines that enhance T-cell mediated immune responses. The correlation of positively selected amino acids (PS) of HIV-1 with host HLA alleles can identify regions containing potential T-cell epitopes. However, the specific epitopes have to be identified and characterized using overlapping peptides through T-cell functional assays. In this study we used a new approach to identify and characterize potential epitopes in the gag region containing PS mutations that significantly correlated with HLA-A*0301. The iTopia Epitope Discovery System was used to rapidly screen a panel of peptides overlapping the regions containing PS mutations and the peptides identified were assessed for relative affinity and complex stability. The potential epitopes were then validated by interferon gamma (IFN-gamma) ELISpot assays with patient PBMCs. Using this approach we identified/confirmed the predicted HLA-A*0301 epitopes in two regions of gag containing PS mutations V7I and K403R, one previously reported and the other novel. Five of the seven peptides that bound to A*0301 contained the K403R mutation and corresponded to the documented LARNCRAPRK-A3 supertype epitope. Two epitope variants, RASVLSGGK and RASILSGGK containing the V7I mutation, were identified using the iTopia Epitope Discovery System, however only the consensus variant (RAK9C) was confirmed using the ELISpot assay and it represents a novel A*0301 epitope. PMID- 19903486 TI - Predict potential drug targets from the ion channel proteins based on SVM. AB - The identification of molecular targets is a critical step in the drug discovery and development process. Ion channel proteins represent highly attractive drug targets implicated in a diverse range of disorders, in particular in the cardiovascular and central nervous systems. Due to the limits of experimental technique and low-throughput nature of patch-clamp electrophysiology, they remain a target class waiting to be exploited. In our study, we combined three types of protein features, primary sequence, secondary structure and subcellular localization to predict potential drug targets from ion channel proteins applying classical support vector machine (SVM) method. In addition, our prediction comprised two stages. In stage 1, we predicted ion channel target proteins based on whole-genome target protein characteristics. Firstly, we performed feature selection by Mann-Whitney U test, then made predictions to identify potential ion channel targets by SVM and designed a new evaluating indicator Q to prioritize results. In stage 2, we made a prediction based on known ion channel target protein characteristics. Genetic algorithm was used to select features and SVM was used to predict ion channel targets. Then, we integrated results of two stages, and found that five ion channel proteins appeared in both prediction results including CGMP-gated cation channel beta subunit and Gamma-aminobutyric acid receptor subunit alpha-5, etc., and four of which were relative to some nerve diseases. It suggests that these five proteins are potential targets for drug discovery and our prediction strategies are effective. PMID- 19903487 TI - The Burrows-Wheeler similarity distribution between biological sequences based on Burrows-Wheeler transform. AB - This work aims at the similarity of biological sequences. Based on the Burrows Wheeler transform, a definition of Burrows-Wheeler similarity distribution of two sequences is proposed to compare two sequences. Some distance measures are naturally followed by the distribution. The expectation and entropy of the similarity distribution are used to construct phylogenetic trees on two independent data sets. The result demonstrates that the method is efficient and powerful. PMID- 19903488 TI - Dosing-time dependent effect of dexamethasone on bone density in rats. AB - AIMS: While glucocorticoids are widely used to treat patients with various diseases, they often cause adverse effects such as bone fractures. In this study, we investigated whether the decrease in bone density induced by glucocorticoid therapy was ameliorated by optimizing a dosing-time. MAIN METHODS: Rats were administered with dexamethasone (Dex) orally (1mg/kg/day) for 6weeks at a resting or an active period. After the end of the treatment, bone density of femur, biomarkers of bone formation and resorption, and other biomedical variables were measured. KEY FINDINGS: Bone density of femur was significantly decreased by the 6-week treatment with Dex, and the degree of decrease in the 14 HALO (hours after light on) dosing group (an active period) was larger than that in the 2 HALO dosing group (a resting period). Although urinary calcium excretion was accelerated by Dex treatment, secondary hyperparathyroidism was not detected. Histomorphometry analysis showed that Dex suppressed bone resorption, which was larger in the 2 HALO than in the 14 HALO groups. These data indicate that Dex equally suppressed bone formation in the 2 and 14 HALO groups, but inhibited bone resorption more in the 2 HALO than in the 14 HALO groups. SIGNIFICANCE: This study shows that the decrease in bone density induced by Dex was changed by its dosing-time. PMID- 19903489 TI - Beneficial effects of thymoquinone on hepatic key enzymes in streptozotocin nicotinamide induced diabetic rats. AB - AIMS: The present study was designed to evaluate the antihyperglycemic potential of thymoquinone (TQ), major constituent of Nigella sativa seeds on the activities of key enzymes of carbohydrate metabolism in streptozotocin (STZ)-nicotinamide (NA)-induced diabetic rats. MAIN METHODS: Diabetes was induced in experimental rats weighing 180-220g, by a single intraperitoneal (i.p) injection of STZ (45mg/kg b.w), 15min after the i.p administration of NA (110mg/kg b.w). Diabetic rats were administered TQ intragastrically at 20, 40, 80mg/kg b.w for 45days. The levels of plasma glucose, insulin, glycated hemoglobin (HbA(1C)) and hemoglobin (Hb) were measured. The activities of hexokinase, glucose 6-phosphate dehydrogenase, glucose 6-phosphatase and fructose 1,6-bisphosphatase were assayed in liver homogenates. KEY FINDINGS: Oral administration of TQ for 45days, dose dependently improved the glycemic status in STZ-NA induced diabetic rats. The levels of insulin, Hb increased with significant decrease in glucose and HbA(1C) levels. The altered activities of carbohydrate metabolic enzymes were restored to near normal. No significant changes were noticed in normal rats treated with TQ. SIGNIFICANCE: These results show that TQ at 80mg/kg b.w is associated with beneficial changes in hepatic enzyme activities and thereby exerts potential antihyperglycemic effects. PMID- 19903492 TI - An antibody surface for selective neuronal cell attachment. AB - An optimal surface for culturing human embryonic stem cell (hESC)-derived neuronal cells is of high interest. In this study, a specific antibody to a neural cell adhesion molecule (NCAM) was immobilised on a solid surface of polystyrene and used as a selective matrix for culturing of hESC-derived neuronal cells. Thereafter, hESC-derived neurospheres were seeded on the matrix. The neurospheres did not attach to the NCAM antibody containing matrix whereas individual neuronal cells did. The neuronal cell attachment was depended on the NCAM antibody concentration. The neuronal cells were viable on the NCAM antibody containing matrix during an 8 day follow-up and exhibited typical bipolar morphology of immature neurons. Specific binding of the NCAM antigen to an immunoglobulin-polymer coated surface was verified by surface plasmon resonance (SPR) measurements. This study is to our knowledge the first demonstrating the use of an antibody layer as a selective surface for hESC-derived neuronal cells. PMID- 19903490 TI - Sex shapes experimental ischemic brain injury. AB - Biologic sex and sex steroids are important factors in clinical and experimental stroke. This review evaluates key evidence that biological sex strongly alters mechanisms and outcomes from cerebral ischemia. The role of androgens in male stroke is understudied and important to pursue given that male sex is a well known risk factor for human stroke. To date, male sex steroids remain largely evaluated at the bench rather than the bedside. We review recent advances in our understanding of androgens in the context of ischemic cell death and neuroprotection. We also highlight some possible molecular mechanisms by which androgens impact ischemic outcomes. PMID- 19903493 TI - Differentiation of mouse Neuro 2A cells into dopamine neurons. AB - Neuro 2A (N2a) is a mouse neural crest-derived cell line that has been extensively used to study neuronal differentiation, axonal growth and signaling pathways. A convenient characteristic of these cells is their ability to differentiate into neurons within a few days. However, most differentiation methods reported for N2a cells do not provide information about the neuronal types obtained after each treatment. In this study, we evaluated the generation of N2a dopamine neurons following treatment with a number of factors known to induce neuronal differentiation. Our results showed that N2a cells express Nurr related factor 1 (Nurr1) and produce low levels of tyrosine hydroxylase (TH) and dopamine. Both TH and dopamine levels were significantly enhanced in the presence of dibutyryl cyclic adenosine monophosphate (dbcAMP), as evidenced by Western blot, immunocytochemistry and high performance liquid chromatography (HPLC). In contrast to dbcAMP, other factors such as transforming growth factor beta1 (TGF beta 1), bone morphogenetic protein 4 (BMP4), glial cell-derived neurotrophic factor (GDNF) and retinoic acid (RA) did not increase TH expression. Further investigation confirmed that the effect of dbcAMP on production of TH-positive neurons was mediated through cyclic AMP (cAMP) responsive element binding protein (CREB) and it was antagonized by RA. Thus, although various treatments can be used to generate N2a neurons, only dbcAMP significantly enhanced the formation of dopamine neurons. Taken together, this study provided a simple and reliable method to generate dopamine neurons for rapid and efficient physiological and pharmacological assays. PMID- 19903491 TI - Cystic fibrosis: exploiting its genetic basis in the hunt for new therapies. AB - Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), an anion channel expressed in epithelial cells throughout the body. In the lungs, absence or dysfunction of CFTR results in altered epithelial salt and water transport eventuating in impaired mucociliary clearance, chronic infection and inflammation, and tissue damage. CF lung disease is the major cause of morbidity and mortality in CF despite the many therapies aimed at reducing it. However, recent technological advances combined with two decades of research driven by the discovery of the CFTR gene have resulted in the development and clinical testing of novel therapies aimed at the principal underlying defect in CF, thereby ushering in a new age of therapy for CF. PMID- 19903494 TI - A multisystem approach for development and evaluation of inactivated vaccines for Venezuelan equine encephalitis virus (VEEV). AB - A multisystem approach was used to assess the efficiency of several methods for inactivation of Venezuelan equine encephalitis virus (VEEV) vaccine candidates. A combination of diverse assays (plaque, in vitro cytopathology and mouse neurovirulence) was used to verify virus inactivation, along with the use of a specific ELISA to measure retention of VEEV envelope glycoprotein epitopes in the development of several inactivated VEEV candidate vaccines derived from an attenuated strain of VEEV (V3526). Incubation of V3526 aliquots at temperatures in excess of 64 degrees C for periods >30 min inactivated the virus, but substantially reduced VEEV specific monoclonal antibody binding of the inactivated material. In contrast, V3526 treated either with formalin at concentrations of 0.1% or 0.5% (v/v) for 4 or 24 h, or irradiated with 50 kGy gamma radiation rendered the virus non-infectious while retaining significant levels of monoclonal antibody binding. Loss of infectivity of both the formalin inactivated (fV3526) and gamma irradiated (gV3526) preparations was confirmed via five successive blind passages on BHK-21 cells. Similarly, loss of neurovirulence for fV3526 and gV3526 was demonstrated via intracerebral inoculation of suckling BALB/c mice. Excellent protection against subcutaneous challenge with VEEV IA/B Trinidad donkey strain was demonstrated using a two dose immunization regimen with either fV3526 or gV3526. The combination of in vitro and in vivo assays provides a practical approach to optimize manufacturing process parameters for development of other inactivated viral vaccines. PMID- 19903495 TI - Potatovirus X and Tobacco mosaic virus-based vectors compatible with the Gateway cloning system. AB - Virus-based expression vectors are important tools for high-level production of foreign proteins and for gene function analysis through virus induced gene silencing. To exploit further their advantages as fast, high yield replicons, a set of vectors was produced by converting and adapting Potato virus X (PVX) and Tobacco mosaic virus (TMV)-based vectors to allow easy cloning of foreign sequences by the Gateway cloning system. Target genes were cloned efficiently by recombination and successfully expressed in Nicotiana benthamiana following inoculation by Agrobacterium (agroinfection). Using green fluorescent protein (GFP) as marker, high-level expression with both PVX-GW and TMV-GW vectors was confirmed. A Gateway inserted phytoene desaturase gene (pds) fragment in PVX-GW and TMV-GW vectors (PVX-GW-PDS and TMC-GW-PDS), induced gene silencing of the endogenous pds gene in N. benthamiana as evidenced by chlorotic leaves. The PVX GW vector was adapted further by cloning the GFP gene upstream of the Gateway sequences, allowing the easy production of GFP fusions after recombination of a target gene. Subcellular localization of resulting GFP fusion was validated by recombining and expressing the coat protein gene from Tomato chlorotic mottle virus, revealing its nuclear localization. A PVX-GW transient expression assay of a nucleocapsid protein gene fragment of Tomato spotted wilt virus and of a single chain antibody against this protein was shown to confer effective resistance to TSWV infection. PMID- 19903496 TI - Study of Andes virus entry and neutralization using a pseudovirion system. AB - Andes virus (ANDV), a member of the Hantavirus genus in the family Bunyaviridae, causes an acute disease characteristic of New-World hantaviruses called hantavirus pulmonary syndrome (HPS). HPS is a highly pathogenic disease with a case-fatality rate of 40%. ANDV is the only hantavirus reported to spread directly from human-to-human. The aim of the present study was to develop a quantitative and high-throughput pseudovirion assay to study ANDV infection and neutralization in biosafety level 2 facilities (BSL-2). This pseudovirion assay is based on incorporation of ANDV glycoproteins onto replication-defective vesicular stomatitis virus (VSV) cores in which the gene for the surface G protein has been replaced by that encoding Renilla luciferase. Infection by the pseudovirions can be quantified by luciferase activity of infected cell lysates. ANDV pseudovirions were neutralized by ANDV-specific antisera, and there was good concordance between specificity and neutralization titers of ANDV hamster sera as determined by our pseudovirion assay and a commonly used plaque reduction neutralization titer (PRNT) assay. In addition, the pseudovirions were used to evaluate the requirements for ANDV entry, like pH dependency and the role of beta3 integrin, the reported receptor for other pathogenic hantaviruses, on entry. PMID- 19903497 TI - Evaluation of humoral and cellular immune responses against HSV-1 using genetic immunization by filamentous phage particles: a comparative approach to conventional DNA vaccine. AB - Phage display is based on expressing peptides as a fusion to one of the phage coat proteins. To date, many vaccine researches have been conducted to display immunogenic peptides or mimotopes of various pathogens and tumors on the surface of filamentous bacteriophages. In recent years as a new approach to application of phages, recombinant bacteriophage lambda particles were used as DNA delivery vehicles to mammalian cells. In this study, recombinant filamentous phage whole particles were used for vaccination of mice. BALB/c mice were inoculated with filamentous phage particles containing expression cassette of Herpes simplex virus 1 (HSV-1) glycoprotein D that has essential roles in the virus attachment and entry. Both humoral and cellular immune responses were measured in the immunized mice and compared to conventional DNA vaccination. A dose-response relationship was observed in both arms of immune responses induced by recombinant filamentous phage inoculation. The results were similar to those from DNA vaccination. Filamentous phages can be considered as suitable alternative candidate vaccines because of easier and more cost-effective production and purification over plasmid DNA or bacteriophage lambda particles. PMID- 19903498 TI - Changes of substance P during fracture healing in ovariectomized mice. AB - Neuropeptides may play an important role in the healing process of osteoporotic fractures. The objective of this study was to determine the role of substance P during osteoporotic fracture healing. One hundred ninety-two mice were randomized into ovariectomy (OVX) and control (CON) group (n=96, respectively). Femoral shaft fracture was created 3 weeks after OVX. Bone mineral density (BMD), micro CT (microCT) analysis of fracture callus formation and mineralization, microCT analysis of fracture site neovascularization and biomechanical property as well as substance P levels were evaluated 1, 2, 4, and 8 weeks after fracture and compared with CON group. Following OVX-induced bone loss, fracture healing in OVX mice was significantly poorer than that in CON mice, with a significant decrease of substance P at the fracture site at all time points and with the level at early stage (1 and 2 weeks) higher than later stage (4 and 8 weeks). Impaired angiogenesis was also noted in OVX mice. No significant change of substance P level in serum was found between different groups or time points. In conclusion, fracture healing is inferior in OVX-induced bone loss and associated with a significant decrease of substance P. Substance P may play an important role during osteoporotic fracture healing. PMID- 19903499 TI - Peptide therapy with pentadecapeptide BPC 157 in traumatic nerve injury. AB - We focused on the healing of rat transected sciatic nerve and improvement made by stable gastric pentadecapeptide BPC 157 (10 microg, 10ng/kg) applied shortly after injury (i) intraperitoneally/intragastrically/locally, at the site of anastomosis, or after (ii) non-anastomozed nerve tubing (7 mm nerve segment resected) directly into the tube. Improvement was shown clinically (autotomy), microscopically/morphometrically and functionally (EMG, one or two months post injury, walking recovery (sciatic functional index (SFI)) at weekly intervals). BPC 157-rats exhibited faster axonal regeneration: histomorphometrically (improved presentation of neural fascicles, homogeneous regeneration pattern, increased density and size of regenerative fibers, existence of epineural and perineural regeneration, uniform target orientation of regenerative fibers, and higher proportion of neural vs. connective tissue, all fascicles in each nerve showed increased diameter of myelinated fibers, thickness of myelin sheet, number of myelinated fibers per area and myelinated fibers as a percentage of the nerve transected area and the increased blood vessels presentation), electrophysiologically (increased motor action potentials), functionally (improved SFI), the autotomy absent. Thus, BPC 157 markedly improved rat sciatic nerve healing. PMID- 19903500 TI - Stable isotope probing: technical considerations when resolving (15)N-labeled RNA in gradients. AB - RNA based stable isotope probing (SIP) facilitates the detection and identification of active members of microbial populations that are involved in the assimilation of an isotopically labeled compound. (15)N-RNA-SIP is a new method that has been discussed in recent literature but has not yet been tested. Herein, we define the limitations to using (15)N-labeled substrates for SIP and propose modifications to compensate for some of these shortcomings. We have used (15)N-RNA-SIP as a tool for analysing mixed bacterial populations that use nitrogen substrates. After incubating mixed microbial communities with (15)N ammonium chloride or (15)N(2) we assessed the fractionation resolution of (15)N RNA by isopycnic centrifugation in caesium trifluoroacetate (CsTFA) gradients. We found that the more isotopic label incorporated, the further the buoyant density (BD) separation between (15)N- and (14)N-RNA, however it was not possible to resolve the labeled from unlabeled RNA definitively through gradient fractionation. Terminal-restriction fragment length polymorphism (T-RFLP) analysis of the extracted RNA and fluorescent in situ hybridisation (FISH) analysis of the enrichment cultures provided some insight into the organisms involved in nitrogen fixation. This approach is not without its limitations and will require further developments to assess its applicability to other nitrogen fixing environments. PMID- 19903501 TI - Frontal-parietal synchrony in elderly EEG for visual search. AB - Aging involves selective changes in attentional control. However, its precise effect on visual attention is difficult to discern from behavioural studies alone. In this paper, we employ a recently developed phase-locking measure of synchrony as an indicator of top-down/bottom-up control of attention to assess attentional control in the elderly. Fourteen participants (63-74 years) searched for a target item (coloured, oriented rectangular bar) among a display set of distractors. For the feature search condition, where none of the distractors shared a feature with the target, search time did not increase with display set size (two, or four items). For the conjunctive search condition, where each distractor shared either a colour or orientation feature with the target, search time increased with display size. Phase-locking analysis revealed a significant increase in high gamma-band (36-56 Hz) synchrony indicating greater bottom-up control for feature than conjunctive search. In view of our earlier study on younger (21-32 years) adults (Phillips and Takeda, 2009), these results suggest that older participants are more likely to use bottom-up control of attention, possibly triggered by their greater susceptibility to attentional capture, than younger participants. PMID- 19903502 TI - Culture medium induced vimentin reorganization associates with enhanced baculovirus-mediated gene delivery. AB - Baculoviruses can express transgenes under mammalian promoters in a wide range of vertebrate cells. However, the success of transgene expression is dependent on both the appropriate cell type and culture conditions. We studied the mechanism behind the substantial effect of the cell culture medium on efficiency of the baculovirus transduction in different cell lines. We tested six cell culture mediums; the highest transduction efficiency was detected in the presence of RPMI 1640 medium. Vimentin, a major component of type III intermediate filaments, was reorganized in the optimized medium, which associated with enhanced nuclear entry of baculoviruses. Accordingly, the phosphorylation pattern of vimentin was changed in the studied cell lines. These results suggest that vimentin has an important role in baculovirus entry into vertebrate cells. Enhanced gene delivery in the optimized medium was observed also with adenoviruses and lentiviruses. The results highlight the general importance of the culture medium in the assembly of the cytoskeleton network and in viral gene delivery. PMID- 19903504 TI - Enhancing the utility of alanine aminotransferase as a reference standard biomarker for drug-induced liver injury. AB - Drug-induced liver injury (DILI) is the most frequent cause of discontinuation of new chemical entities during development. DILI can either be intrinsic/predictable or an idiosyncratic type. These two forms of DILI are contrasted in their manifestation and diagnosis. Even with regulatory guidance (FDA, 2009), there is still a gap in our ability to identify predictable DILI, both specifically and sensitively. Alanine aminotransferase (ALT) is the principal reference standard biomarker to diagnose DILI, yet its current application in preclinical to clinical translation for decision-making purposes has imperfections: (1) analytical ALT assay uniformity across industry would be aided by common analytical processes; (2) assessment of ALT toxicological performance in a large preclinical analysis would help to establish a true threshold of elevation for predictable DILI and improve translational use across various stages of pharmaceutical development and finally, (3) clinical evaluation of ALT elevations prospectively and retrospectively is recommended to define and manage variations in clinical study subjects including rising body mass index (BMI) range and ALT upper limit of normal (ULN) in the broader population over time. The emergence of new hepatotoxicity biomarkers necessitates a parallel and equivalent assessment to the aminotransferases in a regulatory qualification model. PMID- 19903503 TI - Liposomal modular complexes for simultaneous targeted delivery of bioactive gases and therapeutics. AB - Intrinsically echogenic liposomes (ELIP) can be adapted to encapsulate nitric oxide to facilitate ultrasound-enhanced delivery of therapeutic agents to atherosclerotic plaques. However, the NO loading of targeted ELIP caused a 93% decrease of antibody (Ab) immunoreactivity. The following hypothesis was tested: biotin/avidin-mediated coupling of NO-ELIP and Ab-conjugated ELIP will enable co delivery of bioactive gases and ELIP that can encapsulate other agents without loss of targeting efficiency. Complex formation was initiated by addition of excess streptavidin to equal proportions of biotinylated Ab-ELIP and NO-ELIP. Fluorescence deconvolution microscopy, Coulter Multisizer 3 analysis and flow cytometry demonstrated that the ELIP coupling procedure formed mixed aggregates of >or=10 liposomes within 1 min. Intravascular ultrasound imaging and ELISA showed that echogenicity and targeting efficiency were completely and 69-99% retained, respectively. When complexed to NO-ELIP, ELIP bifunctionally targeted to both CD34 and ICAM-1 (BF-ELIP) increased human mononuclear cell migration through human coronary artery endothelial cell monolayers in transwell plates 4 fold relative to a nonspecific IgG-ELIP control and 2-fold relative to BF-ELIP alone. It was concluded that this novel multi-functional conjugation methodology provides a platform technology for site-specific co-delivery of bioactive gases and other agents. PMID- 19903505 TI - A critical review of chronic stress effects on spatial learning and memory. AB - The purpose of this review is to evaluate the effects of chronic stress on hippocampal-dependent function, based primarily upon studies using young, adult male rodents and spatial navigation tasks. Despite this restriction, variability amongst the findings was evident and how or even whether chronic stress influenced spatial ability depended upon the type of task, the dependent variable measured and how the task was implemented, the type and duration of the stressors, housing conditions of the animals that include accessibility to food and cage mates, and duration from the end of the stress to the start of behavioral assessment. Nonetheless, patterns emerged as follows: For spatial memory, chronic stress impairs spatial reference memory and has transient effects on spatial working memory. For spatial learning, however, chronic stress effects appear to be task-specific: chronic stress impairs spatial learning on appetitively motivated tasks, such as the radial arm maze or holeboard, tasks that evoke relatively mild to low arousal components from fear. But under testing conditions that evoke moderate to strong arousal components from fear, such as during radial arm water maze testing, chronic stress appears to have minimal impairing effects or may even facilitate spatial learning. Chronic stress clearly impacts nearly every brain region and thus, how chronic stress alters hippocampal spatial ability likely depends upon the engagement of other brain structures during behavioral training and testing. PMID- 19903507 TI - A 4-week repeated oral dose toxicity study of fucoidan from the Sporophyll of Undaria pinnatifida in Sprague-Dawley rats. AB - Fucoidan is extracted from brown seaweeds, which can have anti-coagulant, antithrombotic, antitumor, and antiviral activities. However, detailed studies on the toxicology of fucoidan have not been performed. Here we tested the toxicity of fucoidan in Sprague-Dawley rats. Fucoidan (1350mg/kg bw/day for 4 weeks) did not induce statistically significant differences in groups matched by gender with respect to body weight, ophthalmoscopy, urinalysis, hematology, and histopathology. Fucoidan did not change prothrombin time or activated partial thromboplastin time, indicating an inability to change blood clotting. This study demonstrated that fucoidan is not toxic under this administration paradigm. PMID- 19903506 TI - Involvement of the endocannabinoid system in the neurobehavioural effects of stress and glucocorticoids. AB - The endocannabinoid system is a neuroactive lipid signaling system that functions to gate synaptic transmitter release. Accumulating evidence has demonstrated that this system is responsive to modulation by both stress and glucocorticoids within the hypothalamus and limbic structures; however, the nature of this regulation is more complex than initially assumed. The aim of the current review is to summarize the research to date which examines the effects of acute stress and glucocorticoid administration on endocannabinoid signaling in limbic-hypothalamic pituitary-adrenal (LHPA) axis, and in turn the role endocannabinoid signaling plays in the neurobehavioural responses to acute stress and glucocorticoid administration. The majority of research suggests that acute stress produces a mobilization of the endocannabinoid 2-arachidonoylglycerol (2-AG) while concurrently reducing the tissue content of the other endocannabinoid ligand anandamide. Genetic and pharmacological studies demonstrate that the reduction in anandamide signaling may be involved in the initiation of HPA axis activation and the generation of changes in emotional behaviour, while the increase in 2-AG signaling may be involved in terminating the stress response, limiting neuronal activation and contributing to changes in motivated behaviours. Collectively, these studies reveal a complex interplay between endocannabinoids and the HPA axis, and further identify endocannabinoid signaling as a critical regulator of the stress response. PMID- 19903508 TI - Working in partnership to advance the 3Rs in toxicity testing. AB - Toxicological assessment of pharmaceutical and non-pharmaceutical chemicals is a regulatory requirement to ensure all compounds likely to be exposed to humans or the environment are safe. These studies rely on the use of large numbers of animals and involve a number of assumptions and extrapolations that remain controversial in assuring consumer safety. The UK's National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) has taken a collaborative approach to identify opportunities for implementation of the 3Rs principles (Replacement, Reduction and Refinement) to drive innovation and support animal welfare in toxicity testing. This review highlights the mechanisms by which the NC3Rs is working with the pharmaceutical and chemical industries and regulatory authorities to achieve these goals. PMID- 19903509 TI - Relativism, regulation and the dangers of indifferent science: the Sir Roy Cameron lecture of the Royal College of Pathologists. AB - The evaluation of data obtained from scientific investigations is not easy and the process often seems counterintuitive to the uninformed. Some commentators hold the conviction that ideological motives colour all deliberations-this makes it easy to suggest that in any scientific debate an opponent's reason for holding a particular viewpoint or belief depends on his or her motives, rather than their knowledge base. This position may be useful in providing the grounds on which to mount a polemic against any perceived threat (drugs in modern medicine, pesticides in intensive agriculture or genetically modified organisms). However, its existence is not a reason to ignore the fact that the data obtained by use of the scientific method may be of a different kind from that which depends on opinions, honed by prejudices. Here I use prejudice to mean a bias that may be favourable or unfavourable to a particular viewpoint, rather than to a willful disregard of fact or an unreasoning predilection to arrive at a particular answer -the Precautionary Principle is a good example of only considering results that fit a preconceived viewpoint. The conviction that opinions cannot be based on independent thought has led to a disregard of professionalism and the development of the view that who pays you determines your opinion--not your science. This is relativism, which embodies the assertion that knowledge is relative and that ethical truths depend on the individuals and groups holding them-there are no absolutes. This compounds the problems of regulatory toxicology which is always beset by unavoidable uncertainties and suffers from being based on the use of the results of testing procedures many decades old which have themselves failed to incorporate new knowledge about the processes we investigate (see Hartung, 2009). A more vigorous assertion of the particular values of the scientific viewpoint and a willingness to incorporate new knowledge into regulatory decision making is necessary. PMID- 19903510 TI - Curcumin, resveratrol and flavonoids as anti-inflammatory, cyto- and DNA protective dietary compounds. AB - Numerous dietary compounds, ubiquitous in fruits, vegetables and spices have been isolated and evaluated during recent years for their therapeutic potential. These compounds include flavonoid and non-flavonoid polyphenols, which describe beneficial effects against a variety of ailments. The notion that these plant products have health promoting effects emerged because their intake was related to a reduced incidence of cancer, cardiovascular, neurological, respiratory, and age-related diseases. Exposure of the body to a stressful environment challenges cell survival and increases the risk of chronic disease developing. The polyphenols afford protection against various stress-induced toxicities through modulating intercellular cascades which inhibit inflammatory molecule synthesis, the formation of free radicals, nuclear damage and induce antioxidant enzyme expression. These responses have the potential to increase life expectancy. The present review article focuses on curcumin, resveratrol, and flavonoids and seeks to summarize their anti-inflammatory, cytoprotective and DNA-protective properties. PMID- 19903511 TI - Nonverbal transitive inference: Effects of task and awareness on human performance. AB - We studied human nonverbal transitive inference in two paradigms: with choice stimuli orderable along a physical dimension and with non-orderable choice stimuli. We taught 96 participants to discriminate four overlapping pairs of choice stimuli: A+ B-, B+ C-, C+ D-, and D+ E-. Half of the participants were provided with post-choice visual feedback stimuli which were orderable by size; the other half were not provided with orderable feedback stimuli. In later testing, we presented novel choice pairs: BD, AC, AD, AE, BE, and CE. We found that transitive responding depended on task awareness for all participants. Additionally, participants given ordered feedback showed clearer task awareness and stronger transitive responding than did participants not given ordered feedback. Associative models (Wynne, 1995; Siemann and Delius, 1998) failed to predict the increase in transitive responding with increasing awareness. These results suggest that ordered and non-ordered transitive inference tasks support different patterns of performance. PMID- 19903512 TI - Reorienting when cues conflict: A role for information content in spatial learning? AB - In two experiments, human participants searched in dynamic three-dimensional virtual-environment rectangular enclosures. Unlike previous studies involving learning of features and geometry, we trained features and geometry separately before placing them in conflict. Specifically, participants learned to respond to rewarded features located along the principle axis of a rectangular search space and to respond to rewarded geometry of a rectangular search space in separate training phases followed by a single test trial. During the test trial, features and geometry were placed in conflict by situating rewarded bins during feature training in unrewarded geometric corners from geometry training and unrewarded bins during feature training in rewarded geometric corners from geometry training. Results of Experiment 1 indicated that although all participants learned features and geometry at an equivalent rate and to an equivalent level, performance during the test trial indicated no preferential responding to features or geometry. However, choice reaction time was significantly longer during the test trial compared to that of last feature and last geometry training trials. Experiment 2 attempted to dissociate information content of features and geometry from their acquired associative strength by rewarding only one geometric corner during geometry training. Results of Experiment 2 indicated that although features had presumably acquired greater associative strength relative to that of geometry by the end of training, performance during the test trial indicated no preferential responding to features or geometry. As in Experiment 1, choice reaction time was significantly longer during the test trial compared to that of last feature and last geometry training trials. Collectively, results seem to provide converging evidence against a view-based matching account of spatial learning, appear inconsistent with standard associative-based accounts of spatial learning, and suggest that information content of spatial cues may play an important role in spatial learning. PMID- 19903513 TI - Kinetic studies of nitrofurazone photodegradation by multivariate curve resolution applied to UV-spectral data. AB - This work aims at describing the kinetic model of nitrofurazone photodegradation by a novel chemometric technique, hybrid hard-soft multivariate curve resolution (HS-MCR). The study was applied to UV-spectral data from the photolysis of nitrofurazone solutions at different concentrations and exposed under varying illuminance power. The HS-MCR method was able to elucidate the kinetics of the photodegradation process and to determine the rate constants, and estimating at the same time the pure spectra of the degradation products. Exposure to light of the drug gave a first rapid isomerization to the syn-form that in turn underwent degradation furnishing a mixture of yellow-red products. The photodegradation process could be explained with a kinetic model based on three consecutive first order reactions (A>B, B>C and C>D). These results were confirmed by application of the MCR procedure to the analysis of the data obtained from HPLC-DAD analysis of the nitrofurazone samples at different reaction times. The kinetic model was observed to be dependent on experimental conditions. The samples at higher concentrations showed rate constants lower than the diluted samples, whereas an increase of the rate of all degradation processes was observed when the light power also increased. This work shows the power of the hybrid hard- and soft multivariate curve resolution as a method to deeply study degradation processes of photolabile drugs. PMID- 19903514 TI - The Fischer 344 rat as a model of presbycusis. AB - Due to the rising number of the aged human population all over the world, presbycusis is a phenomenon that deserves the increasing attention of the medical community as regards to prevention and treatment. This requires finding appropriate animal models for human presbycusis that will be useful in future experiments. Among the available rat strains, the Fischer 344 (F344) strain promises to serve as a model producing prompt and profound presbycusis. Hearing thresholds begin to increase in this strain during the first year of life; toward the end of the second year, the thresholds are very high. The threshold shifts progress independently in both ears. The rapid deterioration of distortion product otoacoustic emissions, with the majority of outer hair cells (OHC) being present and morphologically intact, is apparently produced by the disruption of prestin. The age-related changes within inner ear function are accompanied by deterioration of acoustical signal processing within central auditory system, mainly due to impaired GABA inhibition. The loss of GABA inhibition in old animals is expressed primarily in the inferior colliculus but is also present in the cochlear nuclei and the auditory cortex. Sound-evoked behavioral reactions are also impaired in old F344 rats. Taken together, the described characteristics of the aging F344 rat auditory system supports the idea that this strain may serve as a suitable model for studying the mechanisms of presbycusis, its prevention and treatment. PMID- 19903515 TI - Potential ex vivo immunomodulatory effects of San-Huang-Xie-Xin-Tang and its component herbs on mice and humans. AB - AIM OF THE STUDY: San-Huang-Xie-Xin-Tang (SHXXT), an important Chinese medicine formula, contains Rhei Rhizoma (RR), Scutellariae Radix (SR) and Coptidis Rhizoma (CR). RR and SR are abundant in anthraquinone and flavonoid polyphenols. Pharmacokinetic study of SHXXT indicated that glucuronides were the predominant forms of polyphenols in rats. MATERIALS AND METHODS: As an extension of pharmacokinetic study, the serum metabolites of SHXXT, RR, SR and CR were prepared from rats and quantitated, then the immunomodulation effects were examined by culturing these serum metabolites with murine and human immune cells. RESULTS: The results indicated that the inhibitions on nitric oxide (NO) and cytokine production from mitogen-activated peritoneal macrophages by the serum metabolites of SHXXT, RR, SR and CR were through reducing the protein expression of inducible NO synthase (iNOS) and the IC(50) were 0.8%, 1.5%, 3.0% and 0.8% of their blood concentrations, respectively. In addition, the serum metabolites of SHXXT, RR, SR and CR significantly decreased the ratios of interferon-gamma (IFN gamma) to interleukin (IL)-4 in mitogen-stimulated mice spleen cells and human peripheral blood mononuclear cells (PBMCs). Moreover, the serum metabolites of SHXXT and SR significantly arrested the mitogen-stimulated mice spleen cells at G2/M stage. CONCLUSIONS: In conclusion, the serum metabolites of SHXXT and the component herbs exerted promising modulation activities on the immune functions and the cell cycle distribution of mice and human immune cells. We suggest that SHXXT is a promising remedy for immunomodulation through Th1/Th2 regulation. PMID- 19903517 TI - Comparison of urodynamic effects of phytoestrogens equol, puerarin and genistein with these of estradiol 17beta in ovariectomized rats. AB - Whether urinary incontinence in the postmenopause can be prevented or delayed by estrogens is currently controversially debated. Ovariectomized (ovx) rats have been successfully used as models in the past years but plant derived substances with estrogenic effects in the lower urinary tract have not been studied so far. Therefore we compared the effects of a 3 months lasting oral administration of estradiol 17beta (E2) with those of the phytoestrogens equol, genistein and puerarin. They were ovariectomized, fed with test substance containing food and then anaesthetized and catheterized with a biluminal catheter having one outlet in the bladder and another in the urethra at the level of the urethral sphincter. Urethral and bladder pressure were recorded during a 240s period of retrograde bladder filling (2 x 0.5 ml within 30s with 1 min filling intermission). Bladder and urethra pressures were highest in the E2>puerarin>equol>genistein treated animals. Phytoestrogen and E2 treatment resulted in consistently higher urethral than the bladder pressures during the filling period and in the filled status whereas bladder often exceeded urethral pressures in ovx controls. In summary, we demonstrate significant improvement of urethral closure mechanism under E2 and phytoestrogen administration that can be assumed to be beneficial for prevention or therapy of postmenopausal urge incontinence. PMID- 19903516 TI - Skeletal muscle autophagy and apoptosis during aging: effects of calorie restriction and life-long exercise. AB - Sarcopenia, loss of muscle mass and function, is a common feature of aging. Oxidative damage and apoptosis are likely underlying factors. Autophagy, a process for the degradation of cellular constituents, may be a mechanism to combat cell damage and death. We investigated the effect of age on autophagy and apoptosis in plantaris muscle of male Fischer 344 rats that were either fed ad libitum, or mild, life-long calorie restricted (CR) alone or combined with life long voluntary exercise. Upstream autophagy-regulatory proteins were either upregulated with age (Beclin-1) or unchanged (Atg7 and 9). LC3 gene and protein expression pattern as well as LAMP-2 gene expression, both downstream regulators of autophagy, however, suggested an age-related decline in autophagic degradation. Atg protein expression and LC3 and LAMP-2 gene expression were improved in CR rats with or without exercise. The age-related increase in oxidative damage and apoptosis were attenuated by the treatments. Both, oxidative damage and apoptosis correlated negatively with autophagy. We conclude that mild CR attenuates the age-related impairment of autophagy in rodent skeletal muscle, which might be one of the mechanisms by which CR attenuates age-related cellular damage and cell death in skeletal muscle in vivo. PMID- 19903518 TI - Magnesium sulfate treatment alters fetal cerebellar gene expression responses to hypoxia. AB - Prenatal perturbation of brain circulation and oxygenation is a leading cause of perinatal brain damage affecting about 0.3-0.9% of births. Hypoxia-ischemia (HI) in preterm human infants at gestational week 23-32 results in neurodevelopmental abnormalities in childhood, presenting as learning disability, seizure activity, motor impairment and in the most severe cases, death. Here, we examined the potential of MgSO4 treatment, prior to foetal hypoxia, to attenuate hypoxia induced damage in a murine model of maternal hypoxia. We studied the time course of maternal hypoxia and MgSO4 pre-treatment effects on cerebellar tissue by means of DNA microarray analyses. Mild hypoxia induced minor expression changes in most genes. However, there were 5 gene sets which were down-regulated by maternal hypoxia. MgSO4 pre-treatment abrogated these decreases in gene. A cell cycle gene set which responded immediately (2 h) to hypoxia, showed a delayed response (24 h) when MgSO4 pre-treatment was given. Similar proportions of cell death were observed in all groups before P7, where combined hypoxia and MgSO4 treatment increased cell death in the internal granule layer. There were a higher number of BrdU positive cells at the end of hypoxic episodes and a down-regulation of Reelin signaling, compared to control. MgSO4 pre-treatment prevented the enhancement of cell proliferation due to hypoxia and increased Reelin levels. Altogether, MgSO4 pre-treatment both reduced the number of genes differentially affected by hypoxia and delayed the responses to hypoxia. In addition, MgSO4 pre treatment modified the nature of the transcriptional response; while hypoxia induced down-regulation of gene sets, MgSO4 pre-treatment mostly up-regulated them. The dual reaction to the MgSO4 treatment may be the source of the ambiguity in observations reported for affected newborns. PMID- 19903519 TI - Microglial MyD88 signaling regulates acute neuronal toxicity of LPS-stimulated microglia in vitro. AB - Although the role of microglial activation in neural injury remains controversial, there is increasing evidence for a detrimental effect in the immature brain, which may occur in response to release of neurotoxic substances including pro-inflammatory cytokines. However, the signaling mechanisms involved in microglial-induced neuronal cell death are unclear. Microglia isolated from the brains of wild-type (WT) or MyD88 knockout (KO) mice were exposed to PBS or the TLR4-ligand LPS (100 ng/mL) for 2, 6, 14, or 24 h, and the microglia conditioned medium (MCM) collected. Detection of multiple inflammatory molecules in MCM was performed using a mouse 22-plex cytokine microbead array kit. Primary neuronal cultures were supplemented with the 14 or 24 h MCM, and the degree of neuronal apoptosis examined after exposure for 24 h. Results showed a rapid and sustained elevation in multiple inflammatory mediators in the MCM of WT microglia exposed to LPS, which was largely inhibited in MyD88 KO microglia. There was a significant increase in apoptotic death measured at 24 h in cultured neurons exposed to CM from either 14 or 24 h LPS-stimulated WT microglia (p<.05 vs. WT control). By contrast, there was no increase in apoptotic death in cultured neurons exposed to CM from 14 or 24 h LPS-stimulated MyD88 KO microglia (p=.15 vs. MyD88 KO control). These data suggest that MyD88-dependent activation of microglia by LPS causes release of factors directly toxic to neurons. PMID- 19903520 TI - JNK inhibition and inflammation after cerebral ischemia. AB - The c-Jun-N-terminal kinase signaling pathway (JNK) is highly activated during ischemia and plays an important role in apoptosis and inflammation. We have previously demonstrated that D-JNKI1, a specific JNK inhibitor, is strongly neuroprotective in animal models of stroke. We presently evaluated if D-JNKI1 modulates post-ischemic inflammation such as the activation and accumulation of microglial cells. Outbred CD1 mice were subjected to 45 min middle cerebral artery occlusion (MCAo). D-JNKI1 (0.1 mg/kg) or vehicle (saline) was administered intravenously 3 h after MCAo onset. Lesion size at 48 h was significantly reduced, from 28.2+/-8.5 mm(3) (n=7) to 13.9+/-6.2 mm(3) in the treated group (n=6). Activation of the JNK pathway (phosphorylation of c-Jun) was observed in neurons as well as in Isolectin B4 positive microglia. We quantified activated microglia (CD11b) by measuring the average intensity of CD11b labelling (infra red emission) within the ischemic tissue. No significant difference was found between groups. Cerebral ischemia was modelled in vitro by subjecting rat organotypic hippocampal slice cultures to oxygen (5%) and glucose deprivation for 30 min. In vitro, D-JNKI1 was found predominantly in NeuN positive neurons of the CA1 region and in few Isolectin B4 positive microglia. Furthermore, 48 h after OGD, microglia were activated whereas resting microglia were found in controls and in D-JNKI1-treated slices. Our study shows that D-JNKI1 reduces the infarct volume 48 h after transient MCAo and does not act on the activation and accumulation of microglia at this time point. In contrast, in vitro data show an indirect effect of D-JNKI1 on the modulation of microglial activation. PMID- 19903521 TI - Social disruption induces lung inflammation. AB - Social disruption (SDR) is a well-characterized mouse stressor that causes changes in immune cell reactivity in response to inflammatory stimuli. In this study, we found that SDR in the absence of an immune challenge induced pulmonary inflammation and increased pulmonary myeloperoxidase activity. The percentage of neutrophils within the lungs increased 2-fold after social disruption. Monocyte accumulation in the lungs was also significantly increased. In addition, SDR increased the percentage of neutrophils that expressed CD11b, indicating that more neutrophils were in an activated state. In the lungs, we observed an increased level of the inflammatory cytokine, IL-1beta, as well as higher levels of KC/CXCL1, MIP-2/CXCL2, and MCP-1/CCL2, which are chemokines responsible for neutrophil and monocyte recruitment. Furthermore, social disruption led to increased lung expression of the adhesion molecules P-selectin, E-selectin, and ICAM-1, which localize and recruit immune cells. These data support previous findings of an inflammatory environment induced by SDR. We demonstrate that this effect also occurs in the pulmonary milieu and in the absence of an inflammatory stimulus. PMID- 19903522 TI - The antioxidative effect of estradiol therapy on erythrocytes in women with preeclampsia. AB - In the present study, we evaluated changes of both oxidative stress marker concentrations in erythrocytes and values of blood pressure, as well as their relation during short-term estradiol therapy in preeclampsia. Serum estradiol concentrations were also recorded. The results of this study showed significant decrease of mean arterial pressure (MAP) values during estradiol therapy, whereas there was no significant change in serum estradiol concentrations. Decreased concentrations of superoxide anion (O(2)(-)), hydrogen peroxide (H(2)O(2)), nitrite (NO(2)(-)), peroxynitrite (ONOO(-)) and lipid peroxide (LPO) were found during estradiol therapy in erythrocytes. No changes were found in the activity of gluthatione-S-transferase (GST). The decrease of MAP values was positively correlated with the reduction of concentrations of O(2)(-), H(2)O(2), NO(2)(-) and ONOO(-) in erythrocytes during estradiol therapy. The obtained results suggest that short-term intramuscular administration of estradiol shows antioxidative effects in erythrocytes and reduces blood pressure in preeclampsia. PMID- 19903523 TI - Evaluation of a modified comet assay to detect DNA damage in mammalian sperm exposed in vitro to different mutagenic compounds. AB - The final stages of male gametogenesis are sensitive targets of DNA-reactive chemicals, most of which form adducts. Comet assay is a widely applied genotoxicity test that reveals DNA adducts through breaks formed during repair processes. However, sperm cells are essentially devoid of repair enzymes and comet assay is poorly sensitive in detecting chemically induced DNA lesions in sperm. To overcome such limitation, in a previous paper we proposed a modified protocol for comet assay. In this work we further tested the method treating bull sperm with additional mutagens (diethylsulfate, mitomycin C, bleomycin and colchicine) in parallel with the standard comet assay. No treatment-related increase of DNA migration was ever detected with the standard protocol. A dose dependent effect of diethylsulfate, was obtained with the modified assay. As expected, the mitotic poison colchicine resulted negative even by the modified assay. Results with the other two compounds were consistent with their mechanism of action. PMID- 19903524 TI - Inhibition of hyaluronidase activity of human and rat spermatozoa in vitro and antispermatogenic activity in rats in vivo by Terminalia chebula, a flavonoid rich plant. AB - Our interest in development of hyaluronidase inhibitors as male antifertility agents led to identification of Terminalia chebula (T. chebula) plant with hyaluronidase (HAase) inhibitory activity of human spermatozoa ( approximately 93% inhibition) and rat caudal epididymal spermatozoa ( approximately 86% inhibition) in vitro at 30 mg/ml. We further demonstrated inhibition of hyaluronidase activity of testis and epididymal spermatozoa in vivo coincident with antispermatogenic activity and contraceptive efficacy of TC extract administered at 50 and 100mg/kg/day orally for 60 days in male albino rats. The significant decrease in motility, count and increase in morphological abnormalities of epididymal spermatozoa and severe reduction in fertility (-100%) of male rats treated with T. chebula fruit extract at 100mg/kg dose could be attributed to either direct effect on testis or direct or indirect interference with sperm maturation in epididymis, and/or inhibition of testicular and epididymal sperm hyaluronidase enzyme in vivo probably caused by flavonoids like tannins present in T. chebula. PMID- 19903525 TI - Reproductive toxicity of acrylamide-treated male rats. AB - Acrylamide content is elevated in fried, baked and heat-processed starchy foods. The present experiment was conducted to investigate the reproductive toxicity of oral acrylamide in male rats. Thirty weaned SD male rats of 21-day-old were randomly allotted to three groups, and acrylamide was administered to each group at doses of 0, 5 and 10 mg/kg-d for 8 consecutive weeks. The results indicated that the growth of rats treated with acrylamide was retarded (P<0.05), but relative weights of testes and epididymides compared to body weight were not significantly different (P>0.05). Our results also indicate that the epididymal sperm reserves decreased significantly (P<0.05), suggesting partial depletion of germ cells. In addition, histopathologic lesions were also present in the testes of treated rats. Furthermore, distinct expression patterns of sGC heterodimers were observed in this animal model. This may suggest different physiologic roles for sGC subunits in spermiogenesis and steroidogenesis. PMID- 19903526 TI - New insights into the mode of action of ultradeformable vesicles using calcein as hydrophilic fluorescent marker. AB - Whether ultradeformable vesicles pass intact through the stratum corneum and can promote the transdermal absorption of any substance remain open questions. This paper presents different experimental approaches, based on the use of calcein as hydrophilic fluorescent marker, to probe the physicochemical and pharmacokinetic characteristics of these vesicles. Ultradeformable membranes made from natural phosphatidylcholine and sodium cholate were found to be highly permeable to calcein, as a result of the permeabilizing effects of sodium cholate and ethanol. In vitro skin permeation and in vivo transdermal (percutaneous) absorption studies were performed using hairless mice. Both studies indicated that deformable vesicles reduce the transdermal flux of calcein, when compared to a solution containing or not sodium cholate and ethanol. The data support the model that the transdermal absorption of calcein from deformable vesicles is controlled by the release of the drug from the formulation deposited onto the skin surface. Importantly, fluorescence measurements of the receptor fluid of the Franz diffusion cell after addition of Co(2+) quencher revealed that permeated calcein exists essentially under the non-encapsulated form. In conclusion, our results argue against the model that deformable vesicles would carry hydrophilic drugs across the skin and act as a sustained release system in deep tissues. PMID- 19903527 TI - Development of swelling/floating gastroretentive drug delivery system based on a combination of hydroxyethyl cellulose and sodium carboxymethyl cellulose for Losartan and its clinical relevance in healthy volunteers with CYP2C9 polymorphism. AB - The aim of this study was to develop an optimal gastroretentive drug delivery system (GRDDS) for administering Losartan. Additionally, the influence of optimized GRDDS on the bioavailability of Losartan and the formation extent of active metabolite E3174 by CYP2C9 polymorphism was investigated. Swellable and floatable GRDDS tablets combining hydroxyethyl cellulose (HEC), sodium carboxymethyl cellulose (NaCMC), and sodium bicarbonate were prepared at various compression pressures for evaluating swelling characteristics and floating capacity. Then Losartan was incorporated into optimized formulations for in vitro and in vivo characterizations. An appropriate ratio of HEC to NaCMC, addition of sodium bicarbonate, and compression at lower pressures resulted in the tablets floating over SGF for more than 16 h and swelling to 2 cm in diameter within 3h. The release patterns of Losartan from these tablets were pH-dependent. Results of the clinical trials showed that the mean bioavailability from GRD-A (HEC 91.67%, sodium bicarbonate 3.33% and Losartan 8.33%) was approximately 164%, relative to the immediate-release product (Cozaar). MRT and t(max) values were greater and C(max) values were lower for the GRDDS tablets compared with Cozaa. The lower bioavailability of Losartan in the CYP2C9*1/*1 subjects than CYP2C9*1/*3 subjects was found and could be due to the variety of enzymatic activity. PMID- 19903528 TI - An Adaptive Resonance Theory account of the implicit learning of orthographic word forms. AB - An Adaptive Resonance Theory (ART) network was trained to identify unique orthographic word forms. Each word input to the model was represented as an unordered set of ordered letter pairs (open bigrams) that implement a flexible prelexical orthographic code. The network learned to map this prelexical orthographic code onto unique word representations (orthographic word forms). The network was trained on a realistic corpus of reading textbooks used in French primary schools. The amount of training was strictly identical to children's exposure to reading material from grade 1 to grade 5. Network performance was examined at each grade level. Adjustment of the learning and vigilance parameters of the network allowed us to reproduce the developmental growth of word identification performance seen in children. The network exhibited a word frequency effect and was found to be sensitive to the order of presentation of word inputs, particularly with low frequency words. These words were better learned with a randomized presentation order compared with the order of presentation in the school books. These results open up interesting perspectives for the application of ART networks in the study of the dynamics of learning to read. PMID- 19903529 TI - Two-dimensional crystallization conditions of human leukotriene C4 synthase requiring adjustment of a particularly large combination of specific parameters. AB - Human leukotriene C(4) synthase (LTC(4)S) forms highly ordered two-dimensional (2D) crystals under specific reconstitution conditions. It was found that control of a larger number of parameters than is usually observed for 2D crystallization of membrane proteins was necessary to induce crystal formation of LTC(4)S. Here, we describe the parameters that were optimized to yield large and well-ordered 2D crystals of LTC(4)S. Careful fractioning of eluates during the protein purification was essential for obtaining crystals. While the lipid-to-protein ratio was critical in obtaining order, four parameters were decisive in inducing growth of crystals that were up to several microns in size. To obtain a favorable diameter, salt, temperature, glycerol, and initial detergent concentration had to be controlled with great care. Interestingly, several crystal forms could be grown, namely the plane group symmetries of p2, p3, p312, and two different unit cell sizes of plane group symmetry p321. PMID- 19903531 TI - Phylogeny of ladybirds (Coleoptera: Coccinellidae): are the subfamilies monophyletic? AB - The Coccinellidae (ladybirds) is a highly speciose family of the Coleoptera. Ladybirds are well known because of their use as biocontrol agents, and are the subject of many ecological studies. However, little is known about phylogenetic relationships of the Coccinellidae, and a precise evolutionary framework is needed for the family. This paper provides the first phylogenetic reconstruction of the relationships within the Coccinellidae based on analysis of five genes: the 18S and 28S rRNA nuclear genes and the mitochondrial 12S, 16S rRNA and cytochrome oxidase subunit I (COI) genes. The phylogenetic relationships of 67 terminal taxa, representative of all the subfamilies of the Coccinellidae (61 species, 37 genera), and relevant outgroups, were reconstructed using multiple approaches, including Bayesian inference with partitioning strategies. The recovered phylogenies are congruent and show that the Coccinellinae is monophyletic but the Coccidulinae, Epilachninae, Scymninae and Chilocorinae are paraphyletic. The tribe Chilocorini is identified as the sister-group of the Coccinellinae for the first time. PMID- 19903532 TI - Are lowland rainforests really evolutionary museums? Phylogeography of the green hylia (Hylia prasina) in the Afrotropics. AB - A recent trend in the literature highlights the special role that tropical montane regions and habitat transitions peripheral to large blocks of lowland rainforest play in the diversification process. The emerging view is one of lowland rainforests as evolutionary 'museums'; where biotic diversity is maintained over evolutionary time, and additional diversity is accrued from peripheral areas, but where there has been little recent diversification. This leads to the prediction of genetic diversity without geographic structure in widespread taxa. Here, I assess the notion of the lowland rainforest 'museum' with a phylogeographic study of the green hylia (Aves: Sylviidae: Hylia prasina) using 1132 bp of mtDNA sequence data. The distribution of genetic diversity within the mainland subspecies of Hylia reveals five highly divergent haplotype groups distributed in accordance with broad-scale areas of endemism in the Afrotropics. This pattern of genetic diversity within a currently described subspecies refutes the characterization of lowland forests as evolutionary museums. If the pattern of geographic variation in Hylia occurs broadly in widespread rainforest species, conservation policy makers may need to rethink their priorities for conservation in the Afrotropics. PMID- 19903530 TI - Radiation damage effects at four specimen temperatures from 4 to 100 K. AB - Radiation damage is the primary factor that limits resolution in electron cryo microscopy (cryo-EM) of frozen-hydrated biological samples. Negative effects of radiation damage are attenuated by cooling specimens to cryogenic temperatures using liquid nitrogen or liquid helium. We have examined the relationship between specimen temperature and radiation damage across a broad spectrum of resolution by analyzing images of frozen-hydrated catalase crystal at four specimen temperatures: 4, 25, 42, and 100K. For each temperature, "exposure series" were collected consisting of consecutive images of the same area of sample, each with 10 e(-)/A(2) exposure per image. Radiation damage effects were evaluated by examining the correlation between cumulative exposure and normalized amplitudes or IQ values of Bragg peaks across a broad range of resolution (4.0-173.5A). Results indicate that for sub-nanometer resolution, liquid nitrogen specimen temperature (100K) provides the most consistent high-quality data while yielding statistically equivalent protection from radiation damage compared to the three lower temperatures. At lower resolution, suitable for tomography, intermediate temperatures (25 or 42K) may provide a modest improvement in cryo-protection without introducing deleterious effects evident at 4 K. PMID- 19903533 TI - Phylogenetic relationships in the genus Cyprinella (Actinopterygii: Cyprinidae) based on mitochondrial and nuclear gene sequences. AB - Members of the genus Cyprinella are abundant and broadly distributed cyprinids in central and eastern rivers of North America. Currently, 30 species are placed in this genus, including seven species restricted to Mexico and three barbeled species formerly placed in different genera (Hybopsis, Erimonax). Phylogenetic analyses were performed for all recognized species of Cyprinella plus species from 14 related genera using complete nucleotide sequences of one mitochondrial (cytochrome b) and one nuclear (Rag1) gene. Cyprinella revealed high levels of genetic divergence between species in both genes compared with species from other related genera examined. Analyses of mitochondrial sequences from 98 specimens of Cyprinella revealed the gene trees of some species as non-monophyletic (i.e., C. venusta, C. lepida, C. nivea, C. lutrensis, C. whipplei). Mitochondrial analyses recovered nine well-supported species groups within a main clade inclusive of most species of Cyprinella, although resolution regarding the monophyly of the genus or its relationships with other genera was poorly resolved, except for C. callistia and C. monacha, two species that are apparently not part of the genus. Analyses of nuclear gene sequences from selected distinctive mitochondrial clades also recovered these nine clades and resulted in shorter branches but greater resolution at more basal nodes. Nuclear data also rejected the inclusion of C. monacha in Cyprinella and indicated that Cyprinella is not monophyletic if Tampichthys and Codoma are recognized as valid genera. A major Clade A (formed by Cyprinella, Tampichthys, Codoma) is more closely related to Pimephales, Opsopoeodus, Hybognathus and some Notropis than to Luxilus and Lythrurus as previously hypothesized. Although mitochondrial data are highly informative phylogenetically, our research revealed fundamental conflicts among mitochondrial and nuclear data for several species of Cyprinella. Interestingly, nuclear trees recovered some species resolved as polyphyletic with mitochondrial variation as monophyletic. While we remain highly supportive of phylogenetic studies using mitochondrial genes and genomes, this study does highlight the utility of nuclear data by identifying issues of possible hybridization or retained ancestral polymorphisms and lineage sorting that can yield erroneous hypothesis in studies using only mit-DNA analyses, while shedding invaluable light on the phylogeny of this controversial group of shiners. PMID- 19903534 TI - Quantitative assessment of the metabolic activation of alicyclic amines via aldehyde. AB - INTRODUCTION: Recently it has been reported that some drugs that produce reactive intermediates may cause clinical adverse effects following covalent binding to biomacromolecules. For example, Schiff base production mediated by aldehyde is a possible mechanism of drug-protein adducts. However, because thiols do not trap aliphatic aldehydes via hemiacetal or hemiaminal, the glutathione-trapping method cannot be used to determine the covalent bindings of the Schiff base. METHODS: We established a quantitative method to determine covalent binding mediated by aldehydes via hemiaminal or hemiacetal using non-radiolabeled compound and [(14)C]semicarbazide as a hard-trap agent with unique post-incubation. RESULTS: The trapped aldehyde obtained from the post-incubation was almost equivalent to the covalent binding of the radiolabeled tool compound. Our novel method showed its usefulness in quantitative detection of aldehyde's covalent binding ability by several reagents with alicyclic amine and launched drugs as control. DISCUSSION: The post-incubation method is useful for screening newly synthesized compounds to quantitatively assess the bioactivation of aldehydes descending from alicyclic amines. PMID- 19903535 TI - A simple orchidometric method for the preliminary assessment of maturity status in male cynomolgus monkeys (Macaca fascicularis) used for nonclinical safety studies. AB - INTRODUCTION: The identification and use of mature male non-human primates in nonclinical toxicology studies could be important for evaluating candidate drugs for which the profile of toxicity may differ depending on sexual maturity. This investigation sought to establish operational criteria to complement the current standard of histological evaluation for defining sexual maturity in male cynomolgus monkeys (Macaca fascicularis) used for toxicology studies, and to identify a practical non-invasive measure to select mature males for study. METHOD: Retrospectively, the relationships between body weight, testicular weight and testis histology were established in control males (n=126) used in previous toxicology studies. Prospectively, testicular volumes were measured in-life by orchidometry using comparative scrotal palpation (n=23 males used for study), then compared to testicular weights measured at necropsy. RESULTS: Consistent with previous literature, a weak relationship was observed between body weight and testicular weight. There was, however, a very good relationship between testicular weight and histological maturation level, which was based upon microscopic examination of testes, epididymides and prostates. Orchidometric measurement of testicular volume was found to be a reasonable predictor of testicular weight and served to rapidly select sexually mature males for study, and a total testicular volume (left and right combined) of >20 ml correlated with the histological appearance of maturity. CONCLUSION: Based upon this preliminary exploratory study, the initial simple measurement of testicular volume by orchidometry may provide a non-invasive alternative approach for assessing the sexual maturity of male cynomolgus monkeys in research colonies or during toxicology studies that will require more thorough validation. PMID- 19903536 TI - A lactose specific lectin from the sponge Cinachyrella apion: purification, characterization, N-terminal sequences alignment and agglutinating activity on Leishmania promastigotes. AB - Crude extract from the sponge Cinachyrella apion showed cross-reactivity with the polyclonal antibody IgG anti-CvL (Cliona varians lectin) and also a strong haemagglutinating activity towards human erythrocytes of all ABO groups. Thus, it was submitted to acetone fractionation, IgG anti-deglycosylated CvL Sepharose affinity chromatography, and Fast Protein Liquid Chromatography (FPLC-AKTA Purifier) gel filtration on a Superose 6 10/300 column to purify a novel lectin. C. apion lectin (CaL) agglutinated all types of human erythrocytes with preference for papainized type A erythrocytes. The haemagglutinating activity is independent of Ca2+, Mg2+ and Mn2+ ions, and it was strongly inhibited by the disaccharide lactose, up to a minimum concentration of 6.25 mM. CaL molecular mass, determined by FPLC-gel filtration on a Superose 12 10/300 column and SDS gel electrophoresis, was approximately 124 kDa, consisting of eight subunits of 15.5 kDa, assembled by hydrophobic interactions. The lectin was heat-stable between 0 and 60 degrees C and pH-stable. The N-terminal amino acid sequence of CaL was also determined and a blast search on amino acid sequences revealed that the protein showed similarity only with a silicatein. Leishmania chagasi promastigotes were agglutinated by CaL and this activity was abolished by lactose, indicating that lactose receptors could be presented in this parasite stage. These findings are indicative of the potential biotechnological application of CaL as diagnostic of pathogenic protozoa. PMID- 19903537 TI - Syphacia obvelata modifies mitogen-activated protein kinases and nitric oxide synthases expression in murine bone marrow cells. AB - Syphacia obvelata is a rodent nematode parasite with high prevalence in laboratory mice. In our previous work we have demonstrated that this gut-dwelling helminth induces significant hematopoietic changes, characterized by increased myelopoiesis and erythropoiesis in infected animals, and accompanied with altered reactivity of bone marrow hematopoietic progenitors to interleukin (IL)-17. In this study we extended these investigations by demonstrating that naturally acquired S. obvelata infection induces significant alterations in murine bone marrow cells manifested at the molecular level. Namely, S. obvelata infection induced sustained phosphorylation of the members of three major groups of distinctly regulated mitogen-activated protein kinases (MAPKs), the p38, the c Jun amino-terminal kinase (JNK) and the extracellular signal-regulated kinase (ERK), as well as enhanced expression of mRNA for the inducible nitric oxide synthase (iNOS) in the bone marrow cells of infected animals. Furthermore, the infection interfered with the IL-17-mediated effects in bone marrow cells, since in normal mice IL-17 significantly enhanced phosphorylation of p38 MAPK and upregulated the expression of iNOS and the constitutive, endothelial (e)NOS mRNA, while in S. obvelata-infected animals IL-17 did not influence the MAPKs activation, but markedly down-regulated the expression of both NOS isoforms. The data obtained demonstrating that S. obvelata is able to manipulate signal transduction pathways in the hosts' bone marrow cells, pointed to the multiple layers of immunomodulatory ability of this pinworm parasite and highlighted the importance of working under pinworm-free conditions when using experimental murine models for immunohematopoietic investigations. PMID- 19903538 TI - Genetics vs. entropy: longevity factors suppress the NF-kappaB-driven entropic aging process. AB - Molecular studies in model organisms have identified potent longevity genes which can delay the aging process and extend the lifespan. Longevity factors promote stress resistance and cellular survival. It seems that the aging process itself is not genetically programmed but a random process involving the loss of molecular fidelity and subsequent accumulation of waste products. This age related increase in cellular entropy is compatible with the disposable soma theory of aging. A large array of host defence systems has been linked to the NF kappaB system which is an ancient signaling pathway specialized to host defence, e.g. functioning in immune system. Emerging evidence demonstrates that the NF kappaB system is activated during aging. Oxidative stress and DNA damage increase with aging and elicit a sustained activation of the NF-kappaB system which has negative consequences, e.g. chronic inflammatory response, increase in apoptotic resistance, decline in autophagic cleansing, and tissue atrophy, i.e. processes that enhance the aging process. We will discuss the role of NF-kappaB system in the pro-aging signaling and will emphasize that several longevity factors seem to be inhibitors of NF-kappaB signaling and in that way they can suppress the NF kappaB-driven entropic host defence catastrophe. PMID- 19903539 TI - The geoepidemiology of Sjogren's syndrome. AB - Sjogren's syndrome (SS) is a slowly progressing autoimmune disease, affecting predominantly middle-aged women, with a female to male ratio reaching 9:1. It is characterized by lymphocytic infiltration of the exocrine glands, mainly the lacrimal and salivary glands, resulting in reduced secretory functions and oral and ocular dryness. The syndrome can present alone as primary SS (pSS) or in the context of underlying connective tissue disease as secondary SS (sSS). While the pathogenesis of the disease remains elusive, environmental, genetic and hormonal contributors seem to be involved. Over the last years, compelling evidence has suggested a pivotal role of the epithelium in orchestrating the immune response in the histopathological lesion of Sjogren's syndrome and the term "autoimmune epithelitis" has been proposed as an etiological term. Although the clinical manifestations of pSS patients are mainly those of an autoimmune exocrinopathy, almost half of patients develop extraglandular disease, which may be manifested either by epithelial lymphocytic invasion of lung, liver, or kidney (resulting in interstitial nephritis) or by skin vasculitis, peripheral neuropathy, glomerulonephritis, and low C4 levels. The latter reflect immune-complex mediated disease and confer increased risk for lymphoma development. PMID- 19903540 TI - The geoepidemiology of autoimmune intestinal diseases. AB - Inflammatory bowel diseases (IBD) are chronic diseases of the intestinal tract which principally are composed of ulcerative colitis (UC) and Crohn's disease (CD). The prevalence and incidence of both forms of IBD have historically been higher in developed countries with decreasing North-South gradient. However, more recent evidence demonstrate changing demographics as countries become more developed and immigration increases from underdeveloped countries to developed countries. Typically these changes are marked by an increase in ulcerative colitis followed by an increase in CD. Thus, most if not all human populations appear to be susceptible to IBD under certain environmental influences. Several hypothesis have been advanced to explain these changing demographics including alterations in the bowel microflora, but direct experimental evidence is lacking in most cases. Celiac disease or gluten-sensitive enteropathy is a related inflammatory condition which is induced in susceptible individuals when exposed to gluten-containing foods. Similarly, the prevalence of celiac disease is increasing as the consumption of gluten-containing foods is increasing worldwide. PMID- 19903542 TI - Carbonated apatites obtained by the hydrolysis of monetite: influence of carbonate content on adhesion and proliferation of MC3T3-E1 osteoblastic cells. AB - The influence of the carbonate content in apatites on the adhesion and the proliferation of MC3T3-E1 osteoblastic cells was investigated. B-type carbonated apatites (DCAps) were prepared by the hydrolysis of monetite (CaHPO(4), DCP) in solutions with a carbonate concentration ranging from 0.001 to 0.075 mol l(-1). Stoichiometric hydroxyapatite (DCAp0) was synthesized in carbonate-free solution. MC3T3-E1 cells were seeded on the compacted DCAps and cell adhesion and proliferation were analysed after 24h and 7 days, respectively, using a MTS assay and fluorescence microscopy. Cell adhesion tends to increase with increasing carbonate content for carbonate contents between 0 and 6.9 wt.% and levels out to an acceptable value (+ or - 50% compared to the control) for carbonate contents between 6.9 and 16.1 wt.%. Only DCAps with a carbonate content equal to or higher than 11% support high cell proliferation comparable to the control. On the latter DCAps, the cells have a spread morphology and form a near-confluent layer. A decrease in charge density and crystallinity at the apatite surface, as well as the formation of more spheroidal crystals with increasing carbonate content, might attribute to changes in composition and three-dimensional structure of the protein adsorption layer and hence to the observed cell behaviour. Consequently, only DCAps with a high carbonate content, mimicking early in vivo mineralization, are possible candidates for bone regeneration. PMID- 19903543 TI - Proteomic signature of muscle atrophy in rainbow trout. AB - Muscle deterioration arises as a physiological response to elevated energetic demands of fish during sexual maturation and spawning. Previously, we used this model to characterize the transcriptomic mechanisms associated with fish muscle degradation and identified potential biological markers of muscle growth and quality. However, transcriptional measurements do not necessarily reflect changes in active mature proteins. Here we report the characterization of proteomic profile in degenerating muscle of rainbow trout in relation to the female reproductive cycle using a LC/MS-based label-free protein quantification method. A total of 146 significantly changed proteins in atrophying muscles (FDR <5%) was identified. Proteins were clustered according to their gene ontology identifiers. Muscle atrophy was associated with decreased abundance in proteins of anaerobic respiration, protein biosynthesis, monooxygenases, follistatins, and myogenin, as well as growth hormone, interleukin-1 and estrogen receptors. In contrast, proteins of MAPK/ERK kinase, glutamine synthetase, transcription factors, Stat3, JunB, Id2, and NFkappaB inhibitor, were greater in atrophying muscle. These changes are discussed in light of the mammalian muscle atrophy paradigm and proposed fish-specific mechanisms of muscle degradation. These data will help identify genes associated with muscle degeneration and superior flesh quality in rainbow trout, facilitating identification of genetic markers for muscle growth and quality. PMID- 19903544 TI - Combining low- and high-energy tandem mass spectra for optimized peptide quantification with isobaric tags. AB - Isobaric tagging, via TMT or iTRAQ, is widely used in quantitative proteomics. To date, tandem mass spectrometric analysis of isobarically-labeled peptides with hybrid ion trap-orbitrap (LTQ-OT) instruments has been mainly carried out with higher-energy C-trap dissociation (HCD) or pulsed q dissociation (PQD). HCD provides good fragmentation of the reporter-ions, but peptide sequence-ion recovery is generally poor compared to collision-induced dissociation (CID). Herein, we describe an approach where CID and HCD spectra are combined. The approach ensures efficiently both identification and relative quantification of proteins. Tandem mass tags (TMTs) were used to label digests of human plasma and LC-MS/MS was performed with an LTQ-OT instrument. Different HCD collision energies were tested. The benefits to use CID and HCD with respect to HCD alone were demonstrated in terms of number of identifications, subsequent number of quantifiable proteins, and quantification accuracy. A program was developed to merge the peptide sequence-ion m/z range from CID spectra and the reporter-ion m/z range from HCD spectra, and alternatively to separate both spectral data into different files. As parallel CID in the LTQ almost doesn't affect the analysis duty cycle, the procedure should become a standard for quantitative analyses of proteins with isobaric tagging using LTQ-OT instruments. PMID- 19903546 TI - Preface. PMID- 19903545 TI - Immunization of cattle with a combination of purified intimin-531, EspA and Tir significantly reduces shedding of Escherichia coli O157:H7 following oral challenge. AB - Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a human pathogen that can cause gastrointestinal disease with potentially fatal consequences as a result of systemic Shiga toxin activity. Cattle are the main reservoir host of EHEC O157 and interventions need to be developed that prevent cattle colonization or limit shedding of the organism from this host. EHEC O157 predominately colonizes the bovine terminal rectum and requires a type III secretion system (T3SS) for adherence and persistence at this site. A vaccine based on concentrated bacterial supernatant that contains T3S proteins has shown some efficacy. Here we have demonstrated that vaccination with a combination of antigens associated with T3S mediated adherence; the translocon filament protein, EspA, the extracellular region of the outer membrane adhesin, intimin, and the translocated intimin receptor (Tir) significantly reduced shedding of EHEC O157 from experimentally infected animals. Furthermore, this protection may be augmented by addition of H7 flagellin to the vaccine preparation that has been previously demonstrated to be partially protective in cattle. Protection correlates with systemic and mucosal antibody responses to the defined antigens and validates the targeting of these colonization factors. PMID- 19903547 TI - Chapter 1 - Tubular liposomes with variable permeability for reconstitution of FtsZ rings. AB - We have developed a system for producing tubular multilamellar liposomes that incorporate the protein FtsZ on the inside. We start with a mixture of spherical multilamellar liposomes with FtsZ initially on the outside. Shearing forces generated by applying a coverslip most likely distort some of the spherical liposomes into a tubular shape, and causes some to leak and incorporate FtsZ inside. We describe protocols for liposome preparation, and for preparing membrane-targeted FtsZ that can assemble contractile Z rings inside the tubular liposomes. We also describe the characterization of the multilamellar liposomes in terms of the permeability or leakiness for a small fluorescent dye and larger protein molecules. These liposomes may be useful for reconstitution of other biological systems. PMID- 19903548 TI - Chapter 2 - Detection and analysis of protein synthesis and RNA replication in giant liposomes. AB - In living cells, biochemical reaction systems are enclosed in small lipidic compartments. To experimentally simulate various biochemical reactions occurring in extant cells, giant liposomes are used to reconstruct an artificial model cell. We present methods for conducting a protein synthesis reaction, followed by the reaction catalyzed by the synthesized proteins inside liposomes, and for measurement of the in liposome reaction using a fluorescence-activated cell sorter (FACS). These techniques enable us to perform detailed analysis of the biochemical reactions occurring in the microcompartments, and have the potential to reveal the role of compartmentalization in cellular systems. PMID- 19903549 TI - Chapter 3 - Construction of cell-sized liposomes encapsulating actin and actin cross-linking proteins. AB - To shed light on the mechanism underlying the active morphogenesis of living cells in relation to the organization of internal cytoskeletal networks, the development of new methodologies to construct artificial cell models is crucial. Here, we describe the successful construction of cell-sized liposomes entrapping cytoskeletal proteins. We discuss experimental protocols to prepare giant liposomes encapsulating desired amounts of actin and cross-linking proteins including molecular motor proteins, such as fascin, alpha-actinin, filamin, myosin-I isolated from brush border (BBMI), and heavy meromyosin (HMM). Subfragment 1 (S-1) is also studied in comparison to HMM, where S-1 and HMM are single-headed and double-headed derivatives of conventional myosin (myosin-II), respectively. In the absence of cross-linking proteins, actin filaments (F-actin) are distributed homogeneously without any order within the liposomes. In contrast, when actin is encapsulated together with an actin-cross-linking protein, mesh structures emerge that are similar to those in living motile cells. Optical microscopic observations on the active morphological changes of the liposomes are reported. PMID- 19903550 TI - Chapter 4 - Reconstitution of membrane budding with unilamellar vesicles. AB - Enveloped virus particles select their lipid-protein components and egress by budding from the host cell membranes. The matrix protein of many enveloped viruses has been proposed as a crucial element for viral budding; however, molecular mechanisms behind membrane remodeling by the matrix protein are yet to be unraveled. Here, we describe a set of in vitro functional reconstitution assays that allow quantitative evaluation of both, membrane binding and creation of membrane curvature by the matrix protein isolated from Newcastle Disease Virus. Individual budding events orchestrated by the matrix protein can be resolved in real time. The assays may be applied for direct reconstitution of the on-membrane action of cellular proteins involved in membrane curvature induction upon binding in vivo. PMID- 19903551 TI - Chapter 5 - Detection of antimycolic acid antibodies by liposomal biosensors. AB - Antibodies to mycolic acid (MA) antigens can be detected as surrogate markers of active tuberculosis (TB) with evanescent field biosensors where the lipid antigens are encapsulated in liposomes. Standard immunoassay such as ELISA, where the lipid antigen is not encapsulated, but directly adsorbed to the well-bottoms of microtiter plates, does not yield the required sensitivity and specificity for accurate diagnosis of TB. One reason for this is the cross-reactivity of natural anticholesterol antibodies with MAs. MAs are the major cell wall lipids of mycobacteria. Mycobacterial MA has immunomodulatory properties and elicits specific antibodies in TB patients. Liposomes were optimized for their use as carriers both for the presentation of immobilized purified mycobacterial MA on sensor surfaces, and as a soluble inhibitor of antibody binding in inhibition assays. By using an inhibition assay in the biosensor, the interference by anticholesterol antibodies is reduced. Here, we describe the MA carrying capacity of liposomes with and without cholesterol as a stabilizing agent, optimized concentration and size of liposomes for use in the biosensor assay, comparison of the methods for wave-guide and surface plasmon resonance biosensors and how the cholesteroid nature of MA can be demonstrated by the biosensor when Amphotericin B is allowed to bind to MA in liposomes. PMID- 19903552 TI - Chapter 6 - Solid lipid nanoparticle formulations pharmacokinetic and biopharmaceutical aspects in drug delivery. AB - Solid lipid nanoparticles (SLNs) have emerged as important tools to modify the release profile for a large number of drugs including protein and peptide molecules. SLNs are produced from biocompatible and biodegradable lipid materials, making them a promising therapeutic strategy for drug targeting and delivery, and surmounting the inherent limitations of regulation acceptance. Due to their versatility in loading both lipophilic and hydrophilic molecules in the solid lipid matrix, SLNs depict the ability to prolong, extend or sustain the release profile of the loaded molecules, therefore reducing the repeated administration, and increasing the therapeutic value of a certain treatment. Additional advantages include reduction of drug toxicity and increase of drug bioavailability. To develop SLN formulations for drug targeting and delivery, a basic pharmacokinetic understanding of drug distribution is of major relevance, as well as the biopharmaceutical aspects of the administration route. This chapter provides a fundamental understanding of the pharmacokinetic properties of SLNs, which influence both biopharmaceutical and clinical profiles of the loaded molecules. PMID- 19903553 TI - Chapter 7 - Preparation of complexes of liposomes with gold nanoparticles. AB - Liposomes have been widely used as drug carriers. Visible liposomes have recently become more attractive as drug carriers in personalized medicine. Gold nanoparticles (Au NPs) have unique size- and shape-dependent properties based on their surface plasmon resonance. They can be visualized by computed tomography (CT) and laser optoacoustic imaging. In addition, their photothermogenic properties are useful for photothermal therapy and photoresponsive drug release from liposomes. Therefore, complexation of liposomes with Au NPs is of considerable interest. There are three types of complex: Liposomes containing Au NPs in the inner phase, liposomes with Au NPs at the lipid membrane, and liposomes modified with Au NPs on the surface. This chapter focuses on the preparation and characterization of the third type of complex that is prepared by direct mixing of a Au NP dispersion with a liposome suspension. PMID- 19903554 TI - Chapter 8 - Bio-nanocapsule-liposome conjugates for in vivo pinpoint drug and gene delivery. AB - A bio-nanocapsule (BNC) is an ~50-nm hepatitis B virus (HBV) subviral particle comprising HBV envelope L proteins and a lipid bilayer, and is synthesized in recombinant Saccharomyces cerevisiae. When BNCs are administered intravenously in a mouse xenograft model, they can accumulate specifically in human liver-derived tissues and enter cells efficiently by the HBV-derived human liver-specific infection machinery, localized at the outer-membrane pre-S region of the L protein. BNC specificity for the human liver can be altered to other tissues by substituting the pre-S region using targeting molecules (e.g., antibodies, lectins, cytokines). BNCs can spontaneously form complexes with liposomes (LPs) by the membrane fusogenic activity of the pre-S region. LPs containing various therapeutic materials (e.g., chemicals, proteins, DNA, RNA) can therefore be covered with BNCs to form an ~150-nm BNC-LP conjugate. BNC-LP conjugates injected intravenously can deliver incorporated materials to target tissues specifically and efficiently by utilizing the HBV-derived infection machinery. The stability of BNC-LP conjugates in the blood circulation is similar to that of PEGylated LPs. In this chapter, we describe the preparation and in vivo application of BNC LP conjugates, and the potential of BNC-LP conjugates as in vivo pinpoint drug delivery systems. PMID- 19903555 TI - Chapter 9 - Nanoliposomal dry powder formulations. AB - Liposomal dry powder formulations (DPFs) have proven their superiority over conventional DPFs due to favorably improved pharmacokinetics and pharmacodynamics of entrapped drugs, and thus, reduced local and systemic toxicities. Nanoliposomal DPFs (NLDPFs) provide stable, high aerosolization efficiency to deep lung, prolonged drug release, slow systemic dilution, and avoid macrophage uptake of encapsulated drug by carrier-based delivery of nano-range liposomes. This chapter describes methods of preparation of nanoliposomes (NLs) and NLDPFs, using various techniques, and their characterization with respect to size distribution, flow behavior, in vitro drug release profile, lung deposition, cellular uptake and cytotoxicity, and in vivo pharmacokinetics and pharmacodynamics. Some examples have been detailed for better understanding of the methods of preparation and evaluation of NLDPFs by investigators. PMID- 19903556 TI - Chapter 10 - Lanthanide-loaded paramagnetic liposomes as switchable magnetically oriented nanovesicles. AB - Magnetically oriented liposomes can be prepared by exposing unilamellar spherical systems loaded with paramagnetic lanthanide(III) complexes to hyperosmotic stress. The resulting nonspherical, lens-shaped, nanoparticles can orient within a static magnetic field, depending on the magnetic properties of their membrane bilayer. The orientation of the vesicles can be easily determined by measuring the paramagnetic contribution to the (1)H chemical shift of the intraliposomal water proton resonance. As the latter shift is dominated by the bulk magnetic susceptibility contribution, its sign (negative or positive) reports about the preferred orientation adopted by the nanovesicles. The alignment within the field depends upon the magnetic susceptibility anisotropy of the liposome membrane, Delta(chi)(LIPO). When Delta(chi)(LIPO) has a negative value (e.g., for nonspherical liposomes made of conventional phospholipids), the nanoparticles align with their long axis parallel to the field, whereas when Delta(chi)(LIPO)>0 the vesicles flip by 90 degrees . The sign of the chemical shift of the intraliposomal water resonance is positive in the former case and negative in the latter, respectively. The liposome orientation can be switched by incorporating in the liposome bilayer suitable amphiphilic paramagnetic lanthanide(III) complexes. The sign of Delta(chi)(LIPO), and consequently the magnetic alignment, will correspond to the sign of the magnetic susceptibility anisotropy of the metal complex. The magnetic susceptibility anisotropy is dependent on both the electronic configuration of the lanthanide ion and the structural characteristics of the amphiphilic complex incorporated in the liposome membrane. The magnetic orientation of such vesicles is maintained in vivo, thus opening promising perspectives for the application of nonspherical liposomes in medical imaging. PMID- 19903558 TI - Chapter 12 - DNA-controlled assembly of liposomes in diagnostics. AB - DNA-encoding of solid nanoparticles requires surface chemistry, which is often tedious and not generally applicable. In the presented method, noncovalent attachment of DNA is used to assemble soft nanoparticles (liposomes) in solution. This process displays remarkably sharp thermal transitions from the assembled to disassembled state, thus enabling easy and fast detection of polynucleotides (e.g., DNA or RNA), including single nucleotide polymorphisms (SNPs). PMID- 19903557 TI - Chapter 11 - Reconstitution of membrane proteins in phospholipid bilayer nanodiscs. AB - Self-assembled phospholipid bilayer Nanodiscs have become an important and versatile tool among model membrane systems to functionally reconstitute membrane proteins. Nanodiscs consist of lipid domains encased within an engineered derivative of apolipoprotein A-1 scaffold proteins, which can be tailored to yield homogeneous preparations of disks with different diameters, and with epitope tags for exploitation in various purification strategies. A critical aspect of the self-assembly of target membranes into Nanodiscs lies in the optimization of the lipid:protein ratio. Here we describe strategies for performing this optimization and provide examples for reconstituting bacteriorhodopsin as a trimer, rhodopsin, and functionally active P-glycoprotein. Together, these demonstrate the versatility of Nanodisc technology for preparing monodisperse samples of membrane proteins of wide-ranging structure. PMID- 19903559 TI - Chapter 13 - Soft hybrid nanostructures composed of phospholipid liposomes decorated with oligonucleotides. AB - This chapter reports on the design, preparation, and characterization of liposomes decorated with synthetic lipid-oligonucleotide conjugates. Several key parameters should be considered for a successful preparation of these functional nanostructures that can be employed further as building blocks in DNA-directed assembly of nano-objects. These parameters are reviewed explicitly in this report and their contributions are discussed. PMID- 19903560 TI - Chapter 14 - Synthesis, characterization, and optical response of gold nanoshells used to trigger release from liposomes. AB - Liposomes show great promise as intravenous drug delivery vehicles, but it is often difficult to combine stability in the circulation with rapid, targeted release at the site of interest. Targeting to specific tissues requires developing highly specific ligands with strong affinities to receptors overexpressed on diseased cells; a new cellular target requires developing new ligands and identifying new receptors. Novel photoactivated, hollow, gold nanoshell (HGN)/liposome composites provide a new approach to both controlled release and specific targeting. HGN are extremely efficient near infrared (NIR) light absorbers, and are not susceptible to photobleaching like conventional dyes. Near-complete liposome contents release can be initiated within seconds by irradiating HGNs with an NIR pulsed laser. Targeting the drug is limited only by the dimensions of the laser beam; no specific ligands or antibodies are required, so different tissues and cells can be targeted with the same HGN/liposomes. HGNs can be encapsulated within liposomes or tethered to the outer surface of liposomes for the most efficient drug release. HGNs in liposome solutions can also trigger release, but with lower efficiency. Drug release is induced by adsorbing femto- to nanosecond NIR light pulses that cause the HGNs to rapidly increase in temperature. The resulting large temperature gradients lead to the formation of vapor microbubbles in aqueous solutions, similar to the cavitation bubbles induced by sonication. The collapse of the unstable vapor bubbles causes liposome-membrane rupture and contents release, with minimal damage to the surroundings, and little overall heating of the solution. PMID- 19903561 TI - Chapter 15 - Complex nanotube-liposome networks. AB - Surfactant nanotube-vesicle networks (NVN) belong to the smallest artificial devices known to date for performing controlled chemical operations with enzymes. Newly established means for transport of chemical reactants between containers, as well as advancements in initiation and control of chemical reactions in such systems have opened pathways to new devices with a resolution down to the single molecule level. Here, we summarize the fabrication and functionalization of complex nanotube-liposome networks for such devices, and discuss related aspects of their application for studying chemical kinetics and materials transport phenomena in ultrasmall-scale biomimetic environments. PMID- 19903562 TI - Chapter 16 - Bionanotubules formed from liposomes. AB - Bionanotubules are lipid-bound cylindrical structures with typical diameters in the tens of nanometers and length than can span up to hundreds of micrometers. Besides being observed in nature, bionanotubules can be prepared synthetically by various methods, some of which involve the extension of these structures from lipid vesicles. We describe the formation of lipid nanotubules from liposomes prepared with various lipid mixtures including phosphatidylcholine, phosphatidic acid, and various fluorescent phospholipids. We depict the methods used to extend bionanotubules from surface-attached vesicles, using electric fields as the driving force for bilayer extension and tubular growth. These methods include liposome preparation, surface attachment, and tubular extension by applying modest electric fields (<30V/cm). Methods in which lipid tubules are extended from liposomes that are free in solution and subject to higher magnitude fields are also described. In addition, we summarize other protocols of bionanotubule formation from liposomes, including various modes of micromechanical manipulation of lipid vesicles. PMID- 19903563 TI - Chapter 17 - Engineering cationic liposome siRNA complexes for in vitro and in vivo delivery. AB - RNA interference, the sequence-specific silencing of gene expression by introduction of short interfering RNA (siRNA) is a powerful tool that that the potential to act as a therapeutic agent and the advantage of decreasing toxic effects on normal tissue sometimes seen with conventional treatments i.e. small molecule inhibitors. Naked, unmodified siRNA is poorly taken up by cells and is subject to degradation when exposed to blood proteins during systemic administration. It has also been shown to produce non-specific immune response as well as having the potential to generate 'off-target' effects. Therefore there is a requirement for a delivery system to not only protect the siRNA and facilitate its uptake, but additionally to offer the potential for targeted delivery with an aim of exploiting the high specificity afforded by RNA interference. Cationic liposomes are the most studied, non-viral delivery system used for nucleic acid delivery. As such, the use of cationic liposomes is promising for siRNA for delivery. Furthermore, polyethylene glycol (PEG) can be incorporated into the liposome formulation to create sterically stabilized or 'stealth' liposomes. Addition of PEG can reduce recognition by the reticuloendothelial system (RES) thereby prolonging circulation time. Here we describe a methodology for the complexation of siRNA with cationic liposomes and PEGylated liposomes using two protocols: mixing and encapsulation. Moreover, the different formulations are compared head to head to demonstrate their efficacy for gene silencing. PMID- 19903564 TI - Look but don't touch: Tactile disadvantage in processing modality-specific words. AB - Recent neuroimaging research has shown that perceptual and conceptual processing share a common, modality-specific neural substrate, while work on modality switching costs suggests that they share some of the same attentional mechanisms. In three experiments, we employed a modality detection task that displayed modality-specific object properties (e.g., unimodal shrill, warm, crimson, or bimodal jagged, fluffy) for extremely short display times and asked participants to judge whether each property corresponded to a particular target modality (e.g., auditory, gustatory, tactile, olfactory, visual). Results show that perceptual and conceptual processing share a tactile disadvantage: people are less accurate in detecting expected information regarding the sense of touch than any other modality. These findings support embodied assertions that the conceptual system uses the perceptual system for the purposes of representation. We suggest that the tactile disadvantage emerges for linguistic stimuli due to the evolutionary adaptation of endogenous attention to incoming sensory stimuli. PMID- 19903565 TI - Satellite panicum mosaic virus coat protein enhances the performance of plant virus gene vectors. AB - The coat protein of satellite panicum mosaic virus (SPCP) is known to effectively protect its cognate RNA from deleterious events, and here, we tested its stabilizing potential for heterologous virus-based gene vectors in planta. In support of this, a Potato virus X (PVX) vector carrying the SPMV capsid protein (PVX-SPCP) gene was stable for at least three serial systemic passages through Nicotiana benthamiana. To test the effect of SPCP in trans, PVX-SPCP was co inoculated onto N. benthamiana together with a Tomato bushy stunt virus (TBSV) vector carrying a green fluorescent protein (GFP) gene that normally does not support systemic GFP expression. In contrast, co-inoculation of TBSV-GFP plus PVX SPCP resulted in GFP accumulation and concomitant green fluorescent spots in upper, non-inoculated leaves in a temperature-responsive manner. These results suggest that the multifaceted SPMV CP has intriguing effects on virus-host interactions that surface in heterologous systems. PMID- 19903566 TI - Mercury concentration in meconium and risk assessment of fish consumption among pregnant women in Taiwan. AB - Meconium is a matrix that can be obtained easily and noninvasively and is useful for detecting antenatal fetal exposure to environmental toxins. Taiwan is an island with high fish consumption, and many pregnant women would like to enjoy the benefits of fish without jeopardizing their health or that of their child. The aim of this study is to assess the mercury concentration in meconium in relation to the health risk of mercury exposure. A total of 198 mother-infant pairs residing in the city of HsinChu were recruited for the study between January 2007 and June 2007. The average mean concentration of mercury in meconium was 79.2+/-7.3 ng g(-1) dry wt We use the Monte Carlo technique to assess the uncertainty in risk assessment and the impact of these uncertainties on the estimation of expected risk of mercury intake from fish in mothers. Based on the FAO/WHO's tolerable daily intake of methylmercury (0.23 microg kg(-1)d(-1)), we found that 17.3% and 14.0% of the daily mercury exposure estimated exceeded the reference dose for foreign-born and Taiwan-born mothers, respectively. We found that the mercury concentration in meconium was much higher than in other studies, except for one study done in Tagum in the Philippines where mercury is used in gold mining. This may be because Asia is the largest emitter of anthropogenic mercury, accounting for 53% of worldwide emissions. Sensitivity analysis suggests that mercury concentration in fish and the rate of ingesting fish may be the key parameters for governments offering risk management guidance to protect the health of mothers and unborn babies. PMID- 19903567 TI - Size distribution and chemical composition of airborne particles in south-eastern Finland during different seasons and wildfire episodes in 2006. AB - The inorganic main elements, trace elements and PAHs were determined from selected PM(1), PM(2.5) and PM(10) samples collected at the Nordic background station in Virolahti during different seasons and during the wildfire episodes in 2006. Submicron particles are those most harmful to human beings, as they are able to penetrate deep into the human respiratory system and may cause severe health effects. About 70-80%, of the toxic trace elements, like lead, cadmium, arsenic and nickel, as well as PAH compounds, were found in particles smaller than 1 microm. Furthermore, the main part of the copper, zinc, and vanadium was associated with submicron particles. In practice, all the PAHs found in PM(10) were actually in PM(2.5). For PAHs and trace elements, it is more beneficial to analyse the PM(2.5) or even the PM(1) fraction instead of PM(10), because exclusion of the large particles reduces the need for sample cleaning to minimize the matrix effects during the analysis. During the wildfire episodes, the concentrations of particles smaller than 2.5 microm, as well as those of submicron particles, increased, and also the ratio PM(1)/PM(10) increased to about 50%. On the fire days, the mean potassium concentration was higher in all particle fractions, but ammonium and nitrate concentrations rose only in particles smaller than 1.0 microm. PAH concentrations rose even to the same level as in winter. PMID- 19903568 TI - Effects of zinc on molting and body weight of the estuarine crab Neohelice granulata (Brachyura: Varunidae). AB - The semiterrestrial burrowing crab Neohelice granulata is one of the main inhabitants of the supratidal and intertidal zones of brackish salt marshes, estuaries and coastal lagoons from South America's Atlantic littoral. A large population of this species spreads out Mar Chiquita coastal lagoon (in Argentina) and its corresponding wetlands, and is considered as a key species within this system. Since high values of dissolved heavy metals (including Zn) have been recently reported within Mar Chiquita coastal lagoon, with levels unusually higher than those from other coastal systems within Argentina, it has been explored that the existence of a risk of environmental conditions endanger these populations. So, juveniles of this estuarine crab were experimentally exposed to increasing concentrations of dissolved Zn (i.e., 0, 0.5 and 1 mg Zn(2+)L(-1)) during six months, the time involved between two successive molts; in addition, both the size and weight reached after each molt were also studied in this assay. It can be concluded that zinc can be toxic to crabs only at high concentrations. Considering that levels up to 1 mg ZnL(-1) were recently reported in Mar Chiquita coastal lagoon waters, the potential occurrence of mean chronic effects on the crab population within the coastal lagoon is discussed. PMID- 19903569 TI - Nanomaterials for environmental studies: classification, reference material issues, and strategies for physico-chemical characterisation. AB - NanoImpactNet is a European Commission Framework Programme 7 (FP7) funded project that provides a forum for the discussion of current opinions on nanomaterials in relation to human and environmental issues. In September 2008, in Zurich, a NanoImpactNet environmental workshop focused on three key questions: 1. What properties should be characterised for nanomaterials used in environmental and ecotoxicology studies? 2. What reference materials should be developed for use in environmental and ecotoxicological studies? 3. Is it possible to group different nanomaterials into categories for consideration in environmental studies? Such questions have been, at least partially, addressed by other projects/workshops especially in relation to human health effects. Such projects provide a useful basis on which this workshop was based, but in this particular case these questions were reformulated in order to focus specifically on environmental studies. The workshop participants, through a series of discussion and reflection sessions, generated the conclusions listed below. The physicochemical characterisation information identified as important for environmental studies included measures of aggregation/agglomeration/dispersability, size, dissolution (solubility), surface area, surface charge, surface chemistry/composition, with the assumption that chemical composition would already be known. There is a need to have test materials for ecotoxicology, and several substances are potentially useful, including TiO(2) nanoparticles, polystyrene beads labelled with fluorescent dyes, and silver nanoparticles. Some of these test materials could then be developed into certified reference materials over time. No clear consensus was reached regarding the classification of nanomaterials into categories to aid environmental studies, except that a chemistry-based classification system was a reasonable starting point, with some modifications. It was suggested, that additional work may be required to derive criteria that can be used to generate such categories, that would also include aspects of the material structure and physical behaviour. PMID- 19903570 TI - The importance of source configuration in quantifying footprints of regional atmospheric sulphur deposition. AB - An atmospheric transport-chemistry model is applied to investigate the effects of source configuration in simulating regional sulphur deposition footprints from elevated point sources. Dry and wet depositions of sulphur are calculated for each of the 69 largest point sources in the UK. Deposition contributions for each point source are calculated for 2003, as well as for a 2010 emissions scenario. The 2010 emissions scenario has been chosen to simulate the Gothenburg protocol emission scenario. Point source location is found to be a major driver of the dry/wet deposition ratio for each deposition footprint, with increased precipitation scavenging of SO(x) in hill areas resulting in a larger fraction of the emitted sulphur being deposited within the UK for sources located near these areas. This reduces exported transboundary pollution, but, associated with the occurrence of sensitive soils in hill areas, increases the domestic threat of soil acidification. The simulation of plume rise using individual stack parameters for each point source demonstrates a high sensitivity of SO(2) surface concentration to effective source height. This emphasises the importance of using site-specific information for each major stack, which is rarely included in regional atmospheric pollution models, due to the difficulty in obtaining the required input data. The simulations quantify how the fraction of emitted SO(x) exported from the UK increases with source magnitude, effective source height and easterly location. The modelled reduction in SO(x) emissions, between 2003 and 2010 resulted in a smaller fraction being exported, with the result that the reductions in SO(x) deposition to the UK are less than proportionate to the emission reduction. This non-linearity is associated with a relatively larger fraction of the SO(2) being converted to sulphate aerosol for the 2010 scenario, in the presence of ammonia. The effect results in less-than-proportional UK benefits of reducing in SO(2) emissions, together with greater-than-proportional benefits in reducing export of UK SO(2) emissions. PMID- 19903571 TI - Economic valuation of environmental benefits from wastewater treatment processes: an empirical approach for Spain. AB - Economic research into the design and implementation of policies for the efficient management of water resources has been emphasized by the European Water Framework Directive (Directive 2000/60/EC). The efficient implementation of policies to prevent the degradation and depletion of water resources requires determining their value in social and economic terms and incorporating this information into the decision-making process. A process of wastewater treatment has many associated environmental benefits. However, these benefits are often not calculated because they are not set by the market, due to inadequate property rights, the presence of externalities, and the lack of perfect information. Nevertheless, the valuation of these benefits is necessary to justify a suitable investment policy and a limited number of studies exist on the subject of the economic valuation of environmental benefits. In this paper, we propose a methodology based on the estimation of shadow prices for the pollutants removed in a treatment process. This value represents the environmental benefit (avoided cost) associated with undischarged pollution. This is a pioneering approach to the economic valuation of wastewater treatment. The comparison of these benefits with the internal costs of the treatment process will provide a useful indicator for the feasibility of wastewater treatment projects. PMID- 19903572 TI - Promising novel therapies for the treatment of endometrial cancer. AB - OBJECTIVES: To discuss the novel agents which are being developed for the treatment of advanced and recurrent endometrial carcinoma and to review other molecular targets that may be interesting in the treatment of this disease. While the majority of women with endometrial cancer enjoy a relatively good prognosis, the options for those women who suffer from a disease recurrence are limited and there is a need to identify novel agents. METHODS: A review of clinical trials of novel therapeutic agents and their molecular targets is provided. In addition, a review of the current literature on other potential molecular targets for endometrial cancer was performed. RESULTS: Several phase II trials of novel agents, both alone and in combination with traditional cytotoxic chemotherapy, have been completed or are nearing completion. It appears that the targeted agents may have the most efficacy in combination with cytotoxic chemotherapy or in a multi-targeted agent approach. CONCLUSIONS: Chemotherapy offers the opportunity for a meaningful response rate in women with endometrial cancer, but the responses are often short lived and cure is uncommon in the setting of recurrent disease. The recent increase in molecular targets has led to the availability of many novel therapies. Determining how these agents are to be used, alone or in combination with "standard" therapies, needs to be defined and translational studies are needed to develop rational combinations of these novel agents before we can move into clinical trials. PMID- 19903573 TI - Apples to oranges?: a direct comparison between suicide attempters and suicide completers. AB - BACKGROUND: Suicide attempters and completers may represent different but overlapping groups of distressed individuals. Although depression is related to an increased risk of suicide, the presence of depression may not discriminate suicide attempters from completers. The present study compared suicide attempters and suicide completers on symptoms of depression, the presence of suicide-related variables and stressful life events. AIMS: The present study sought to identify the key differences between 50 suicide attempters and 50 completers, all diagnosed with a Major Depressive Disorder at the time of their suicidal act. METHODS: Suicide attempters and family member informants of suicide completers participated in a thorough psychosocial evaluation. To maximize comparisons with completers, suicide attempters were subclassified based on the lethality of their attempt. RESULTS: Suicide attempters and completers were similar on most measures of depressive symptoms. However, suicide completers were significantly more likely to use alcohol or drugs prior to their suicidal act and they were more likely to leave a suicide note. Suicide completers were significantly more likely to have encountered significant job stress and financial problems. CONCLUSIONS: The present findings have documented several similarities and differences between suicide attempters and suicide completers. Future research may help to clarify the key warning signs that reflect the risk of completed suicide in adults who have been diagnosed with a Major Depressive Disorder. PMID- 19903574 TI - Effectiveness of the extended release formulation of quetiapine as monotherapy for the treatment of acute bipolar depression. AB - BACKGROUND: To evaluate the effectiveness of quetiapine extended release once daily in bipolar depression. METHODS: Double-blind, placebo-controlled study in acutely depressed adults with bipolar I or II disorder, with or without rapid cycling. Patients were randomized to 8 weeks of quetiapine extended release (XR) 300 mg daily monotherapy or placebo. The primary outcome measure was change from baseline to Week 8 in MADRS total score. RESULTS: Quetiapine XR 300 mg once daily (N=133) showed significantly greater improvement in depressive symptoms compared with placebo (N=137) from Week 1 (p<0.001) through to Week 8 (p<0.001). Mean change in MADRS total score at Week 8 was -17.4 in the quetiapine XR group and 11.9 in the placebo group (p<0.001). Response (>or=50 reduction in MADRS total score) and remission (MADRS total score32 and >256 mg/L, respectively. All of the isolates produced 5 extended-spectrum beta-lactamase (ESBLs) with pIs of 5.4 (TEM-1), 6.7 (KPC-2), 8.2 (SHV-12), 8.4 (CTX-M-14), and ArmA 16S rRNA methylase. bla(KPC-2) was located on a large nonconjugative plasmid, whereas armA was located on another conjugative plasmid. Although carbapenem-resistant Enterobacteriaceae remains rare, the emergence of this group of organism merits monitoring. PMID- 19903585 TI - Rapid detection and differentiation of the exfoliative toxin A-producing Staphylococcus aureus strains based on phiETA prophage polymorphisms. AB - The exfoliative toxin A (ETA) is encoded by the gene located on Staphylococcus aureus prophages. We have developed a single-reaction multiplex polymerase chain reaction (PCR) assay for rapid and specific detection of various phiETA prophages of serogroup B responsible for dissemination of eta gene and ETA production in clinical strains. This PCR strategy enabled to classify the ETA-positive strains into 6 groups designated ETA-B1, ETA-B2, ETA-B3, ETA-B4, ETA-B5, and ETA-B6. The method was tested on a diverse set of 101 ETA and/or ETB-positive S. aureus strains isolated in 22 Czech maternity hospitals and 1 Slovak maternity hospital between 1998 and 2009. This novel PCR strategy is reliable in the rapid identification of yet undescribed ETA-converting B prophages and differentiation of the closely related ETA-positive strains, and it is a convenient tool for hospital epidermolytic infection control. PMID- 19903586 TI - Surface gene mutations of hepatitis B virus among high-risk patients with occult hepatitis B virus infection. AB - Surface gene mutants of hepatitis B virus (HBV) have been reported in a variety of patient groups. Because of limited data regarding these mutations in patients with occult HBV infections; we aimed to determine these mutations among high-risk patients with occult HBV infection. The presence of HBV-DNA was determined in patients with isolated anti-HBc by real-time polymerase chain reaction (PCR). Then, surface gene region was amplified by nested PCR and mutations were analyzed after sequencing. The mutations that resulted in nonfunctional hepatitis B surface antigen (HBsAg) were insertion of single nucleotide in 2 cases, which causes frameshift and single-nucleotide replacement, and premature stop codons at Leu15 and Gly10 in the other 2 cases. Amino acid substitution at amino acid position 207(S207N) was found in the other isolates. Our study suggested that "a" region mutations did not play a major role in HBsAg detection, and other genetic and nongenetic factors may be responsible for failure to detect HBsAg by routine laboratory tests. PMID- 19903588 TI - High concentrations of resistin in the peripheral blood of patients with acute basal ganglia hemorrhage are associated with poor outcome. AB - PURPOSE: Resistin increases in peripheral blood of patients with intracerebral hemorrhage (ICH). We sought to evaluate its relation with disease outcome. MATERIALS AND METHODS: Thirty healthy controls and 86 patients with acute ICH were included. Plasma samples were obtained on admission. Its concentration was measured by enzyme-linked immunosorbent assay. RESULTS: Thirty-two patients (37.2%) died from ICH in a week. The plasma resistin level (24.2 +/- 9.7 ng/mL) in patients was significantly higher than that (8.8 +/- 2.4 ng/mL) in healthy controls after adjustment by age, sex, hypertension, diabetes mellitus, hyperlipidemia, and body mass index using analysis of covariate (F = 9.507, P = .003).A univariate correlation analysis found Glasgow Coma Scale (GCS) score and ICH volume, but a multivariate linear regression only selected GCS score (t = 4.587, P < .001) to be related to plasma resistin level. On a multivariate logistic regression, plasma resistin level (odds ratio = 1.257, 95% confidence interval = 1.058-1.492, P = .009) was an independent variable predicting 1-week mortality. A receiver operating characteristic curve identified that a plasma resistin level greater than 26.3 ng/mL predicted 1-week mortality of patients with 81.2% sensitivity and 81.5% specificity (P < .001). Areas under curves of GCS score and ICH volume were not statistically significantly larger than that of plasma resistin level (P > .05). CONCLUSIONS: Increased resistin level is found after ICH, in association with a poor clinical outcome. PMID- 19903587 TI - Trends in antibiotic susceptibility of bloodstream pathogens in hospitalized patients in France, 1996 to 2007. AB - Nationwide surveys of antimicrobial susceptibility of bacteria isolated from bloodstream infections are required to fit empiric therapy to recent trends and detect emerging resistance. We report the results of a French national prospective survey based on the College of Bacteriology-Virology and Hygiene study group network performed each October during the 1996 to 2007 period, with focus on Enterobacteriaceae (7708 isolates) and Staphylococcus aureus (2271 isolates). The most relevant antimicrobial susceptibilities trends were i) a decrease in fluoroquinolones susceptibility among Enterobacteriaceae (96-90%, P < 0.0001) and Escherichia coli isolates (98-89%, P < 0.0001), respectively, ii) the slight but significant decrease in cefotaxime susceptibility among E. coli (P = 0.016), and iii) the significant increase in gentamicin susceptibility among S. aureus strains (P = 0.016). This survey reports antibiotic susceptibility of bloodstream pathogens in France. The empiric use of fluoroquinolones in severe infections should be cautiously monitored by thorough clinical and microbiologic follow-up. PMID- 19903589 TI - Analysis of linkage disequilibrium between the 5' and 3' haplotypes of the beta globin gene cluster in Mexican afromestizos. AB - Analysis of the 5' and 3' haplotypes (Hps) of the beta-globin gene cluster was performed in 110 beta(A) chromosomes from unrelated Mexican afromestizo individuals in order to determine Hardy-Weinberg equilibrium, allelic frequencies, linkage disequilibrium (LD) and association between the 5' and 3' haplotypes. All sites were found to be in Hardy-Weinberg equilibrium (p>0.05). In the whole beta-cluster, only 12.87% of the pairs of loci exhibited significant LD (22/171) (r(2)>0.33). Within the 5'Hp, three pairs of loci were associated (epsilonHincII/(G)gammaHindIII, epsilonHincII/3'psibetaHincII and (G)gammaHindIII/3'psibetaHincII). In the 3'Hp, 19 pairs of loci showed significant LD and were distributed mostly among the -551, -340, Exon1nt6 and IVS2nt16 polymorphisms. The absence of pairs of loci significantly linked between both 5' and 3' Hps is noteworthy. The allelic combinations of the 40 studied polymorphisms (5 sites in the 5' Hp and 35 sites in the 3' Hp) displayed 69 distinct haplotypes, 22 of them belonging to group A, 27 to B, 18 to C and 2 to D, which denoted the great heterogeneity of our population. Further, 1a7A1, 1a7B1 and 1a1C1 were the most common sequences with 8, 9 and 9 chromosomes each. Association analysis between both 5' and 3' Hps revealed strong coherence with the proposed evolutionary histories for the beta-globin gene polymorphisms. 5'Hp1 (+----), which is considered to be an ancestral haplotype, was the most frequent haplotype found in our population and was linked to 24 different sequences in the 3'Hp, demonstrating great heterogeneity. A similar result was found in the 3' Hps, where older alleles (a17A1 and a7B1) were linked to a higher number of 5'Hps. This is the first time that an analysis of association among the 5' and 3' haplotypes and the LD has been performed with 40 polymorphisms distributed in the beta-globin gene cluster in the Mexican afromestizo population. The poor LD observed between and within the 5' and 3' Hps show that this region is very prone to recombination events. PMID- 19903590 TI - How are evoked responses generated? The need for a unified mathematical framework. PMID- 19903592 TI - [Current management of pain in pediatric oncology.]. AB - Pain is a symptom well known by all medical and nursing staff in pediatric oncology. Indeed, pain is inseparable of cancer in children. Pain management is now well codified and integrated into the general supportive care of cancer. Pharmacokinetics of some drugs remains rather unknown in children due to a lack of clinical studies in this specific age-cohort. This in turn sometimes limits the availability of these drugs in pediatric patients. Nevertheless, all efforts are made to design pain management protocols based on data available in adults. This article reviews the current management practices for children with pain. Hard-to-treat benefits more and more from non-pharmacological additional methods and from the use of original molecules, which doubtless tend to further develop. PMID- 19903591 TI - Age-related changes in postural responses revealed by support-surface translations with a long acceleration-deceleration interval. AB - OBJECTIVES: The objectives were to examine (a) whether surface translations with a long, compared to a short, acceleration-deceleration interval could reveal more age-related differences in postural control and (b) whether age-related differences were associated with reactive or anticipatory postural mechanisms. METHODS: Ten older (66-81years) and ten young adults (22-39years) stood on a moveable platform that was unexpectedly translated in the backward direction. Subjects' electromyographic (EMG) and kinematic responses were recorded in response to translations with either a SHORT (100ms) or LONG (2s) acceleration deceleration interval presented in either a predictable or random order. RESULTS: Age-related differences in kinematic postural responses were greater during LONG compared to SHORT translations. However, both LONG and SHORT translations elicited a similar change in EMG latencies and amplitudes between the older and young adults. No age effects on the presentation order (predictable or random) of the translations were observed. CONCLUSIONS: LONG compared to SHORT surface translations magnify the age-related kinematic but not the EMG changes in reactive postural control. The anticipatory component of postural control was not affected by age. SIGNIFICANCE: Translations with longer acceleration-deceleration intervals reveal more age-related differences in postural control, which are otherwise masked by the deceleration effects inherent to shorter translations. PMID- 19903593 TI - [Sedation using ketamine for pain procedures in Pediatric Oncology.]. AB - Procedural sedation and analgesia for children is widely practiced. Since 2005 to 2007, we evaluated the safety and efficacy of ketamine to control pain induced by diagnostic procedures in pediatric oncology patients. Eight hundred fifty procedures were carried out in 125 patients aged 2 to 16 years. We associated EMNO (inhaled equimolar mixture of nitrous oxide and oxygen), atropin (oral or rectal), midazolam (oral or rectal) and ketamin (intravenous). An anesthesiologist injected ketamin. Average dose of ketamine was 0.33 to 2 mg/kg depending on number and invasiveness of procedures. This method requires careful monitoring and proper precautions. With these conditions, no complication was observed. All patients were effectively sedated. These results indicate that ketamine - in association with EMNO, atropine and midazolam - is safe and effective in pain management induced by diagnostic procedures in pediatric oncology patients. The sedative regimen of intravenous ketamine has greatly reduced patient, family and practitioners anxiety for diagnostic and therapeutic procedures. PMID- 19903594 TI - [A positioning pillow to improve lumbar puncture in paediatric haematology oncology patients: a randomized controlled trial.]. AB - ObjectivesLumbar punctures (LPs) are common in children with cancer. Although pain management during the LP has been well standardised, dealing with stress and anxiety is not well addressed yet. Our objective was to evaluate the potential improvement of the LPs success rate using a positioning pillow, to ensure maximum lumbar flexion, and allow paravertebral muscles to relax, in children who are awake, with either conscious sedation or no sedation.Patients and methodsChildren aged 2-18 years undergoing LPs were randomly assigned to a positioning pillow or no intervention. The primary outcome was the rate of success, i.e. achieving the LPs (sampling or injection) at the first attempt, without bleeding (RBC < 50/mm3). The secondary outcomes included: the child's pain, assessed by a self administered visual analogical scales (VAS) for children over 6 years of age; the parents' and caregivers' perception of the child's pain; the satisfaction of the children, the parents, the caregivers and the physician. The child's cooperation and the occurrence of post-LP syndrome were also evaluated.ResultsOne hundred twenty-four children (62 in each group) were included. The LP pillow tended to increase the success rate of LPs (67 vs 57%; P = 0.23), and decreased the post-LP syndromes (15 vs 24%; P = 0.17) but the differences were not statistically significant. In children over 6-year of age (N = 72), the rate of success was significantly higher in the pillow group (58.5 vs 41.5%; P = 0.031), with a tendency to feel less pain (for less pain and better satisfaction) [median VAS: 25 vs 15 mm; P = 0.39] and being more satisfied (84.4 vs 75.0%; P = 0.34).ConclusionEven if the results presented here are not as optimistic as we might have wished, we can confirm that there is a benefit in using this pillow for LPs. It is especially promising in children over 6-year of age. PMID- 19903595 TI - [The "do-not-resuscitate order" in paediatric palliative home care: why should the emergency team be involved ?]. AB - Major challenge in paediatric palliative home care is to anticipate management of future events. In our opinion, one of major approach is to avoid medical futility especially resuscitation attempts in terminally-ill children especially if home care will be organized. We therefore prospectively discussed with proxi what should be attempted (e.g. treat symptoms of pain or discomfort) and what should be avoided for the sake of the child. A crucial part of the discussion included anticipating non resuscitation of the terminally-ill child. We informed in writing local emergency unit coordinator on results of the discussion with care takers and suggested a procedure in case of an emergency call. To include the local emergency unit is now a standard in our paediatric oncology department since two situations may occur: 1) Parental panic while facing difficult terminal symptoms. We recommend that the local emergency unit coordinator dispatches an emergency team to the child's home in order to manage symptoms (seizures, pain, etc.) but avoid any futile resuscitation attempt. Parental decision to maintain the child at home should be re-evaluated regularly. 2) Parents who wish to stay at home as long as possible, refusing home-based death of their terminally-ill child. We recommend that the family doctor decides whether or not to refer the child to the hospital. Emergency team may be called upon based on the child's status and need for medicalised transport. Even if it should be rather rare that parents contact directly the emergency unit and not as usually the home care coordinator, such situation may occur and should be anticipated. Therefore, the anticipation of non-resuscitation recommendations is a key approach in paediatric palliative home care. This complex discussion should not be avoided as parental/medical panic may induce unrealistic requests for futile medical procedures. PMID- 19903596 TI - [Pain in adolescents with cancer.]. AB - Adolescence is a transitional period between childhood and adulthood that can be difficult to cross. The disease is often lived as supplementary suffering, as the word cancer is often linked to a long, painful and lethal disease. The teenager presenting cancer has several obstacles to overcome: his adolescence, his cancer disease and his pain. Pain can be present throughout the cancerous disease: at diagnosis, in case of relapse, or in the event of terminal tumor progression. Pain can be difficult to assess in adolescents who are opposed to treatments or to healthcare. Pain management must be discussed with a multidisciplinary team and has to have a holistic approach including drug therapy and complementary approaches. PMID- 19903597 TI - [Neuropathic pain in cancer.]. AB - Neuropathic pain often occurs in the course of cancer evolution, but also as sequelae of surgical treatment, chemotherapy, or radiotherapy, when a nervous structure is impaired. Clinical features of neuropathic pain are specific, including spontaneous and evoked painful symptoms that are localized in an area of sensitive nerve distribution. Neuropathic pain may be concomitant to nociceptive pain, and its diagnosis can be easily performed in clinical practice using the DN4 questionnaire. Treatment of neuropathic pain is also specific, based on certain antidepressant or antiepileptic, often in combination with strong opioids. PMID- 19903598 TI - [Observational study of the initial prescription of level 3 analgesics in cancer patients.]. AB - To evaluate pain management in cancer patients, a study was conducted examining the treatment circumstances and modalities of initial prescription level 3 analgesics used by 122 French cancer specialists. The rationale for moving to level 3, the implementation and the follow-up were evaluated in 1,038 patients. The reasons underlying the initial prescription were in line with recommendations for clinical practice (WHO, SOR) and the professionals generally preferred molecules with which they were already familiar. Though pain intensity was reduced in 67% of patients, treatment follow-up could have been improved in a number of cases. In particular, titration was not systematically performed, and the interruption of the prescribed treatment (owing to inefficacy or negative side-effects) was not sufficiently timely. The awareness-raising campaigns performed over the past few years should be continued, underlining the importance of early follow-up, notably during the titration phase of level 3 analgesic initiation. PMID- 19903599 TI - [Setting up supportive care in oncology: reflexions and suggestions.]. AB - A group of 19 health professionals implicated in supportive care wanted to suggest some reflexions for organization, setting and evaluation of the supportive care in institutions and health territories. The suggested organization must be applicable to any cancer patient and the place of the care whatever the age, the stage of the disease; in the future, must be applicable to any patient with serious chronic illness. This organization must allow to optimize the accompaniment and the care of the patients and their close relations by 1) precise and regular analysis of their needs; 2) the respect of the continuity of the health care; 3) the setting of collaborative practice and transversality in the care. It is not a new medical speciality but a coordination of competences for patients and their families. PMID- 19903600 TI - [End of chemotherapy decision for metastatic breast cancer patients.]. AB - If chemotherapy beyond the third line often gives sum clinical benefits, it is sometimes prescribed only to avoid telling bad news to patients. Palliative chemotherapy can lead to symptom reduction and greater health related quality-of live, but longer survival is unlikely. Physician's questioning about chemotherapy continuation is an ethical duty so is the discussion with patients and caregivers about prognosis and possibility to receive principally palliative care. Medical information about prognosis must be told "step by step", following patients questioning and their capacity listening to the answers. Exhaustion of chemotherapy efficacy is the best argument to explain chemotherapy stop, which does not mean end-of-life, particularly for patients with slow growing tumour. Maintain hope for patients in regard with their medical situation is vital in a psychological way, to stay alive and to be able to project himself in the close future. Modalities of decisions taking about end of chemotherapy have to be defined but consultation of all caregivers implicated in the palliative patients care is essential. PMID- 19903601 TI - [Psycho-oncology and supportive care: psychiatrist's practice.]. AB - From a psychiatric point of view, supportive care does not only involve coordinating care among caregivers, even though the inadequacy between patients' needs and patients' access to psychological support is a sufficient incentive for promoting these supportive services. In patients with comorbidities, only cooperative efforts between medical and psychological teams can ensure efficient patient management. Supportive care is essentially based on a more systematic assessment of symptoms and of their impact on patient quality of life. Symptoms are not so much a clinical manifestation required to establish the diagnosis of the disease, but they represent the disease itself. They are the guiding principle of patient management, which is based more on "dimensional" than on "categorical" measures, and which requires active participation of the patients through auto-evaluation. Patients try to fit clinical events into a "narrative picture of their disease", which can actually add temporality to the fleeting transiency of symptoms. Exchanges between the different caregivers involved in coordinated supportive care and comparisons between the different versions of the patients' narrative make it possible for the caregivers to elaborate their own understanding of the disease which, somehow, becomes the "validated version". This version will be more or less directly fed back to the patients during the next period of care delivery and will likely interfere with facts, or disorders reported by the patients, thus inducing a distortion in the reported picture. This explicit confrontation between the two "narrative lines" is a potent incentive for the patients to rethink their attitude toward their disease, and to accept the changes sustained by the strong determination of caregivers to improve, even only marginally, their clinical status. The patients are challenged to rethink the events, which, in their narrative, were viewed as omens. There is evidently a psychotherapeutic effect involved. PMID- 19903602 TI - [Requests for active euthanasia: which reality in an oncology center.]. AB - Euthanasia is a controversial issue in today's society. In countries where euthanasia is legal, it is mainly associated with people with cancer. We retrospectively studied the frequency and basis of patients' requests for active euthanasia in the oncology setting.MethodsRecurrent requests for euthanasia made by the patients of Leon-Berard cancer center (Lyon, France) between 2001 and 2003 were recorded by questioning the physicians and nurse supervisors in charge or by collecting information from the minutes of multidisciplinary palliative care meetings. We also collected information on the general health status of the patients, their motives and their evolution over time, as well as responses from caregivers.ResultsWe identified 16 requests for euthanasia. These involved 8 men, 7 women and 1 child (median age, 56 years), corresponding to 1% of the total deaths recorded during the period. In 2 cases, the request had come from the family only. The most frequent motives were psychological distress (38%), desire for self-autonomy (31%) and pain (31%). Half of the patients, particularly those striving for autonomy, persisted with their request until death, whereas 2 of 3 requests motivated by physical or psychological distress were not maintained. Sedation was administered to 3 patients in response to recurrent requests.ConclusionRequests for euthanasia in cancer patients are rare but may occur. Sometimes suffering is not relieved by palliative care and the request is maintained. Dealing with these patients puts caregivers in a difficult situation. PMID- 19903603 TI - [Family members integration in patients' care in haematology units: a new application of proximology.]. AB - Hospitalization per se is a stress for the family members of a loved one. While their role as informal caregivers is well known with at home patients, especially in oncology, they usually are only mere visitors at the hospital. Hence the integration of family members of in patients' care might improve both patients and families' quality of life. Such a supportive care concept has been previously investigated in pediatric, neurology and intensive care units, but there is not such data in hematology although the therapeutic in this field is marked by long term stay in hospitals' rooms. Since 2001, we developed in Clermont-Ferrand University hospital such a project of integration. Put together, the literature data and our first results (opinion poll) show that the main risk of this approach is the exhaustion of family caregivers, their fear of miss conduct and the alteration of the relationship they previously had with the patient. Conversely this approach is likely beneficial in improving both patients and family member's experience during hospitalization in lowering both patients' affective social and may be cultural loneliness and family members' feeling of uselessly and guilt. In France the development of supportive care and therapeutic education in oncology is nowadays encouraged and of there is no legal basis to restrain the development of this concept. Whose concept, nevertheless should be carefully set up and studied and its benefit remain to be clearly demonstrated. PMID- 19903604 TI - Planning for tomorrow, living for today. PMID- 19903605 TI - Recording jaw relationship records: Facebow, stick bites, or both? PMID- 19903606 TI - FDA new drug approvals in 2008. PMID- 19903607 TI - Digital cameras are not all the same! PMID- 19903608 TI - Taking a crown and bridge impression: the three-viscosity method. PMID- 19903609 TI - Effectiveness of resin composite polymerization when cured at different depths and with different curing lights. AB - This study evaluated how photocuring devices affected the microhardness of composite resin cylinders. For this study, 120 specimens of composite were fabricated and allocated randomly into 12 groups (n = 10), according to the light source (quartz-tungsten-halogen (QTH), LED, argon laser, plasma arc) and the height of the specimen (2.0, 4.0, or 6.0 mm). Twenty-four hours after the specimens were fabricated, the Knoop microhardness test was performed on bottom and top surfaces. Statistical analysis showed significant interaction among light sources, between light sources and specimen heights, and between the surfaces. Compared to the QTH specimens, the argon laser and plasma arc specimens showed reduced polymerization on the top surface, while the plasma arc specimens showed reduced polymerization on the bottom surface. The 4.0 mm samples demonstrated higher Knoop microhardness than the 2.0 mm and 6.0 mm samples, especially when argon laser and plasma arc curing lights were used. The microhardness was always higher on the top surface than on the bottom surface. No photocuring unit was able to properly polymerize the bottom surface as completely as the top surface. PMID- 19903610 TI - Repeatability of a dental shade-matching instrument when compared to traditional visual methods of shade evaluation. AB - Electronic shade-matching instruments are available for clinical use; however, their accuracy has not been established. This study evaluated a new electronic method of clinical shade matching compared to the standard visual method. The Vita Easyshade system, an electronic method of shade matching, was used on 40 subjects to measure the central region of each patient's maxillary left central incisor (tooth No. 9). Two visual evaluators with predetermined visual shade matching abilities selected a shade from the same area of tooth No. 9 for all subjects. Student's t-test, using a 95% score confidence interval (CI), was used to compare the two methods. The Vita Easyshade system was accurate 85% of the time in this in vivo study; however, the instrument was predictably accurate only 68-91% of the time at the 95% CI. PMID- 19903611 TI - Longitudinal bond strength evaluation using the deproteinized dentin technique. AB - This study evaluated bond strength to dentin as a result of storage time for conventional adhesive systems (with or without collagen) that had been deproteinized with 10% sodium hypochlorite (NaOCl). For this study, 72 human molars were sectioned in a mesiodistal axial plane and embedded in acrylic resin; at that point, the vestibular and lingual surfaces were worn down with abrasive paper. Acid etching was performed for 15 seconds (using 37% phosphoric acid) and the specimens were divided into 12 groups (n = 6), depending on the adhesive system used, the dentin treatment performed, and the length of evaluation (24 hours or six months). A resin composite was inserted over the prepared area with the aid of a metal matrix. Following a mechanical shear test, fractured surfaces were analyzed by stereomicroscope and the data were submitted to ANOVA and Tukey's test. It was concluded that the dentin deproteinization treatment with 10% NaOCl improved the bond strength in five of the six groups. The bond strength after 24 hours was significantly higher than the bond strength measured after six months. Of the three adhesive systems tested in this study, DenTASTIC UNO demonstrated the lowest bond strength. PMID- 19903612 TI - Portable dental equipment: dental units and x-ray equipment. AB - Portable dental equipment makes it possible for clinicians to provide dental care to patients who are unable to travel to a traditional dental clinic. Nonambulatory, homebound, and institutionalized patients benefit greatly when portable equipment is used to address their dental treatment needs on-site. In recent years, more brands of portable equipment have been introduced to the market, some of which are quite sophisticated in terms of their design and uses. This article seeks to describe two kinds of commercially available portable equipment--dental units and x-ray devices--and to discuss their capabilities and features. PMID- 19903613 TI - Temperature variation caused by high-intensity LED curing lights in bovine dentin. AB - This study sought to evaluate the temperature variations in bovine dentin when cured with high-intensity LED appliances and quartz-tungsten-halogen (QTH) appliances. Forty-five slices of bovine dentin (0.7 mm thick) were divided into three groups. Temperature variations were measured during polymerization of the adhesive (10 seconds), during polymerization of the resin composite (40 seconds), and 24 hours after the resin composite polymerization. The data were submitted to the ANOVA repeated measures test, which showed a statistically significant difference in the interaction factor (p = 0.0001). Tukey's test (p = 5%) revealed that the SmartLite PS LED appliance caused a significantly higher temperature increase than the other appliances following polymerization of the adhesive, that both LED appliances produced significantly greater temperature increases than the QTH curing light during polymerization of the resin composite, and that the SmartLite PS produced the greatest temperature increase after 24 hours. PMID- 19903614 TI - Durability of the bond strength of self-adhesive resin cements to human dentin. AB - This study subjected two self-adhesive resin cements and two conventional resin cements to dry and aging conditions, to compare their microtensile bond strengths (microTBS) to dentin. Using four different luting systems (n = 10), 40 composite resin blocks (each 5 x 5 x 4 mm) were cemented to flat human crown dentin surfaces. The specimens were stored in water for 24 hours (37 degrees C), at which point each specimen was sectioned along two axes to obtain beams that were divided randomly into two groups: dry samples, which were tested immediately, and samples that were subjected to accelerated aging conditions (12,000 thermocycles followed by storage for 150 days). The microTBS results were affected significantly by the luting system used (P < 0.0001). Only the microTBS of Rely-X Unicem was reduced significantly after aging; the microTBS remained stable or increased for the other self-adhesive resin cement and the two conventional cements. PMID- 19903615 TI - An evaluation of dentin conditioners for resin-modified glass ionomer cements. AB - This study sought to test the null hypothesis that there was no difference between three dentin conditioners in terms of how they affected the shear bond strength to dentin of three resin-modified glass ionomer (RMGI) cements. One hundred twenty caries-free third molars were mounted in acrylic resin and their occlusal surfaces were reduced to within 2 mm of their cemento-enamel junction (CEJ). Of the three cements tested, Rely-X Plus (22.7 MPa) and Fuji CEM (19.0 MPa) produced the highest bond strengths when dentin was treated with 37% phosphoric acid and followed by the application of Optibond Solo Plus and BondLink. Among the dentin conditioners tested, Fuji Plus Conditioner increased bond strengths for all three cements more than leaving the smear layer intact, although the increase was not statistically significant. Dentin treated with Cavity Conditioner demonstrated the weakest bond strengths. PMID- 19903616 TI - Initial enamel wear of glazed and polished leucite-based porcelains with different fusing temperatures. AB - This study used the radiotracer method to measure the initial enamel wear caused by low- and high-fusing porcelains after glazing or polishing. It also tested the correlation between enamel wear and porcelain surface roughness (Ra). Surface morphology was assessed by optical microscopy. Cylindrical specimens of three porcelains (two high-fusing, one low-fusing) were either autoglazed or polished. Flattened enamel specimens were irradiated with neutrons and submitted to the wear assay for 2,500 cycles in distilled water using a 285 g load; the released beta 32P particles were measured for 10 minutes. For all samples, Ra was recorded with a profilometer before and after testing. Enamel wear was not significantly different for porcelain or finishing method but there was a trend of interaction between the two variables (p = 0.08). A positive correlation was found between enamel wear and the initial Ra of porcelain (r = 0.71). The glazed surfaces of high-fusing porcelains were wavy and had a greater Ra, while the polished surfaces had grooves and pores prior to wear testing. The low-fusing porcelain demonstrated lower Ra and a more homogeneous surface. All abraded surfaces had similar morphology after the wear assay. PMID- 19903617 TI - Sealing ability of three commercial resin-based endodontic sealers. AB - This study sought to compare the leakage of micro-organisms through root canals that had been obturated using one of three commercial endodontic sealers. Forty maxillary anterior human teeth were randomly assigned to one of three groups, depending on the sealer used. This study utilized an in vitro microbial leakage test with a split chamber design. A mixture of bacterial markers was placed in the upper chamber and could reach the lower chamber (which contained brain heart infusion broth) only by leaking through the root canal filling. Microbial leakage was checked daily for 60 days and data were analyzed statistically by Kruskal Wallis and Mann-Whitney tests. The materials were further evaluated for antimicrobial effect by an agar diffusion test. The results of the leakage test showed no statistically significant difference between Sealer 26 and AH Plus, although both materials differed significantly from Resilon/Epiphany, which took less time to display microbial leakage. All three sealers demonstrated microbial leakage in less than 60 days; however, the agar diffusion test showed that AH Plus and Resilon/Epiphany induced significantly larger microbial inhibition zones than those induced by Sealer 26. PMID- 19903618 TI - Resin bond strength to a zirconia-reinforced ceramic after different surface treatments. AB - This study evaluated the microtensile bond strength between a composite resin and a zirconia-reinforced alumina-based glass-infiltrated ceramic after different ceramic surface treatments. Blocks (12 mm x 10 mm x 5 mm3) of a ceramic containing zirconia were fabricated, polished, and divided at random into seven groups, with each group receiving a different surface treatment. The adhesive system and composite resin were applied to the treated ceramic; these composite ceramic blocks were stored in distilled water (37 degrees C) for seven days. At that point, they were cut along two axes to produce specimens with a cross sectional area of 1.0 (+/- 0.1) mm2. The specimens (n = 20) were loaded in tension in a universal testing machine with a cross-head speed of 0.5 mm/minute. The microtensile bond strength values were calculated and statistically analyzed using ANOVA and Tukey's test (alpha = 0.05). Among the seven groups, specimens with both a silica coating and a silane coupling agent showed the highest mean microtensile bond strength values. All specimens treated with 9.5% hydrofluoric acid failed during the cutting procedure; among all specimens, the majority of the failures were cohesive. A positive correlation was observed between the fracture type and the microtensile bond strength (r = 0.63). PMID- 19903619 TI - Attached gingiva: histology and surgical augmentation. AB - The keratinized attached gingiva provides the periodontium with increased resistance to external injury, contributes to the stabilization of the gingival margin, and aids in dissipating physiological forces exerted by the muscular fibers of the alveolar mucosa on the gingival tissues. Increasing attached gingiva should be strongly considered in cases where the patient's plaque control is compromised. The apically positioned flap, free gingival graft, and subepithelial connective tissue graft are the most common surgical procedures used for augmenting the zone of attached gingiva effectively and predictably. The newly obtained keratinized gingiva can be maintained for a long period; in addition, these periodontal procedures halt the progression of gingival recession and could lead to gaining more keratinized gingiva from creeping attachment after the surgery. This article reviews the biology of attached gingiva and presents cases related to the functional role of periodontal plastic surgery. PMID- 19903620 TI - Recognition and management of odontogenic referred pain. AB - There are times when referred pain makes it difficult to locate the source of odontogenic pain. In those situations, additional evaluation or referral is needed to determine the cause of the chief complaint. This article presents two cases that illustrate the difficulty and frustrations of referred pain for both patients and dentists. After a thorough evaluation and diagnosis, the true sources of the patients' chief complaints were identified and treated, leading to complete resolution of their symptoms. These cases emphasize the importance of listening to the patient, obtaining a detailed dental history, collecting comprehensive diagnostic data, and developing an accurate diagnosis before initiating treatment. When a definitive diagnosis cannot be determined, referral may be necessary to provide the best treatment possible. PMID- 19903621 TI - Vital tooth cleaning for cementation of indirect restorations: a review. AB - This article reviews factors that must be considered to select the best technique for cleaning a vital tooth prior to cementation of a definitive restoration. The dental literature offers many suggestions with supporting rationales. In cases where provisional cement has been present or contrast powder has been used, some mechanical cleaning (with fine pumice or prophy paste) appears to be justified. The value of soaps or other chemicals is questionable except perhaps as topical disinfectants. Dentists should be aware that a cleansing agent may have a negative or positive effect on bond strength, depending on the adhesive system chosen. It may be necessary to tailor the particular method of tooth cleaning to the cement that is to be used. PMID- 19903622 TI - Nonvital tooth bleaching with halogen light-activated agents: case reports and discussion. AB - Esthetic dentistry has received increased attention in recent years, as people are more aware of the esthetic appearance of their teeth, including alignment and whiteness. This development, combined with a decrease in the incidence and severity of caries, has directed some clinicians toward conservative and non invasive treatments such as tooth bleaching. A number of methods for nonvital tooth bleaching are described in the literature; these procedures rely on the bleaching agent used, the agent's concentration, product format, and the source of light activation. This article presents two case reports in which dental bleaching with halogen light-activated agents was used to treat a nonvital discolored incisor. The advantages and disadvantages of the technique are discussed. PMID- 19903623 TI - A case of anterior maxillary radiolucency. AB - A 46-year-old woman sought treatment for pain in her maxillary anterior teeth. Physical examination revealed facial tenderness and an expanded anterior maxilla, with palatal swelling in the approximate midline. The lesion had eroded through the midline anterior palate and anterior nasal spine and had shifted the nasal floor in a superior direction. Teeth No. 8-10 tested nonvital to electrical stimulation. The lesion was dissected carefully and lifted off the nasal floor while the patient was under general anesthesia. Histologically, the lesion was cystic with an epithelial lining composed of respiratory and squamous epithelium. As a result, a nasopalatine duct cyst was diagnosed. PMID- 19903624 TI - Orthodontic implantology: a revolutionary approach to high-anchorage orthodontics. AB - Mini-implants have become a well-accepted standard in orthodontics; as a result, an increasing number of general dentists have seen or will see these anchorage devices in their patients. This article seeks to introduce general dentists to this topic and provide information about the fundamentals of this novel treatment approach. PMID- 19903625 TI - Photodynamic therapy for the treatment of recurrent herpes labialis: preliminary results. AB - This study sought to evaluate the clinical outcome of patients who had been diagnosed with recurrent herpes labialis (RHL) after treatment with photodynamic therapy (PDT) associated with low-level laser therapy (LLLT). PDT has shown great effectiveness for treating already-established RHL vesicles, compared to ordinary treatments involving antiviral compounds. Two patients with vesicles on their lips were treated with PDT, followed by irradiation with LLLT. Both patients reported pain relief immediately after the procedure; at a six-month follow-up, neither patient showed signs or symptoms that related to RHL. PMID- 19903626 TI - A drug-free oral hydrogel wound dressing for pain management in immediate denture patients. AB - This article evaluated a drug-free oral hydrogel wound dressing composed entirely of natural food ingredients for its ability to relieve pain in immediate denture patients. Evaluation occurred at a 24-hour postoperative appointment. For this crossover study, 44 patients who were taking oral narcotics evaluated their discomfort (using a scale of 0-10) at 1, 3, 5, and 10 minutes after denture insertion with no topical treatment and again after SockIt! Oral Pain Gel was applied to the dentures. The gel provided statistically significant pain relief at all time points beyond that provided by oral narcotic alone (p < 0.0001). PMID- 19903627 TI - Cancellous osteoma of the maxillary tuberosity: case report. AB - The osteoma is a benign, bone-forming lesion that is not considered a true neoplasm. It almost exclusively affects the flat bones of the skull and face. Jaw lesions may cause facial deformity and impair oral function. Multiple osteomas may be associated with Gardner's syndrome. Clinically and radiographically, osteomas may need to be differentiated from other bone-forming lesions. This article describes a cancellous osteoma of the left maxillary molar and tuberosity area and differentiates it from other benign and malignant lesions, including the parosteal osteosarcoma. PMID- 19903628 TI - Characterization of oral biopsies from a geriatric population. AB - A retrospective search was performed to find the histological diagnosis for all lesions recorded by an oral pathology service for patients 65 and older from 1969 2002. The search was subcategorized into two age groups and the 20 most common diagnoses from these categories were characterized by patient gender and race. Lesions then were classified into one of seven modified etiopathogenic categories. Of the 52,774 cases recorded during this time, 7,854 involved patients aged 65-84 and 393 cases involved patients 85 and older. Inflammatory lesions were the most common category of lesion in patients over the age of 65. Among patients aged 65-84, fibroma was the most common diagnosis, while squamous cell carcinoma was the most frequent diagnosis among patients 85 and older. Neoplastic lesions (epithelial dysplasia and squamous cell carcinoma) comprised 11.7% of the diagnoses among patients aged 65-84 and 18.6% of the diagnoses among patients 85 and older. These data indicate that patients 85 and older demonstrated a statistically significant increased frequency of squamous cell carcinoma compared to patients in the 65-84 age group. PMID- 19903629 TI - Comparison of techniques to evaluate the quantification of Candida spp. in HIV infected children. AB - This study sought to compare techniques used to make a quantified evaluation of Candida spp. in children infected with HIV. Twenty-four HIV-infected children (age 3 to 13) were selected. Three sterilized swabs were used for each child: one for the dorsum of the tongue, one for the hard palate mucosa, and one for the right jugal mucosa; each swab was rubbed for 10 seconds and transferred to sterilized test tubes containing 1 mL of 0.9% saline solution. Candida spp. growth was observed in 95.8% of all samples, including 95.7% of tongue samples (Group T), 87.0% of saliva samples, 56.6% of hard palate mucosa samples (Group P), and 47.8% of right jugal mucosa samples (Group J). There was no statistical difference in Candida spp. growth between saliva samples and Group T samples, although both had higher growth compared to Groups P and J (p < 0.05; chi(2)). Regarding the sensitivity of each site for positive Candida spp. growth, Group T samples showed 69.5%, while saliva samples showed 52.2%, Group P samples showed 21.7%, and Group J samples showed 13.04%, with no significant statistical difference between Group T and saliva; however, both were more sensitive than Groups J and P (p < 0.05, chi(2)). It was concluded that whole stimulated saliva and swabbing the tongue were considered satisfactory for measuring Candida spp. in HIV-infected children. PMID- 19903630 TI - Management of a horizontal root fracture: a 17-year follow-up. AB - Intra-alveolar root fractures in immature permanent teeth are relatively uncommon and often present a complicated and unpredictable prognosis. This case report documents how conservative treatment was used for the successful management (and long-term follow-up) of immature teeth with a horizontal root fracture. PMID- 19903631 TI - Parallelism of abutment teeth on fixed partial dentures. AB - A total of 100 working dies for fixed partial denture (FPD) abutments were prepared to fit 50 three-unit fixed-fixed conventional FPDs. For each FPD, the axial convergence angles between the mesial and distal axial walls of the abutment teeth were measured. For all FPDs, there were no statistically significant differences between the mesial and distal convergence angles of abutment teeth measured, regardless of their location within the oral cavity. The distal convergence angle of posterior FPD abutments was the highest angle, while the lowest was recorded for the mesial convergence angle of the anterior FPD abutments. PMID- 19903632 TI - Gingival red patch. Juvenile spongiotic gingival hyperplasia. PMID- 19903633 TI - Radiopacity with thin radiolucent rim. Focal cemento-osseous dysplasia. PMID- 19903634 TI - Civics and civility. PMID- 19903635 TI - Aspirin as an emergency drug. PMID- 19903636 TI - A discussion of the physical and oral characteristics of Coffin-Lowry syndrome, Stickler syndrome, and Johanson-Blizzard syndrome. PMID- 19903637 TI - Moving beyond clinical appearance: the need for accurate histological diagnosis. AB - This study considered 120 cases of localized gingival overgrowths that had been clinically diagnosed, surgically excised, and submitted for histopathological examination from 2000 to 2006. Data regarding the patient's age and gender and the lesion's anatomical location, clinical diagnosis, and recurrence were retrieved from case records. The percentage of concurrence and discrepancy between clinical and histopathological diagnosis was calculated using a discrepancy index. In 46.60% of cases, the diagnoses were in total agreement; discrepancies were noted in 53.40% of cases. The diagnosis of peripheral giant cell granuloma showed a high discrepancy (88.89%). Recurrence was seen in five cases. The high discrepancy between the clinical and histopathological diagnosis of localized gingival overgrowths indicates the need for a confirmatory histological examination for diagnosis and complete management of the lesion. PMID- 19903638 TI - Using panoramic radiographs to detect carotid artery calcifications: are they a helpful diagnostic tool? AB - More than 50% of cerebrovascular accidents are caused by atherosclerotic disease at the carotid artery.1 Carotid artery stenosis may be detected by the standard method of duplex ultrasound (DUS), while calcifications can be detected by panoramic radiographs.2-4 Panoramic radiographs may be used as a selective tool, reducing health costs by identifying individuals at risk of stroke or myocardial infarction.5-7 The present study sought a correlation between laboratory findings/behaviors and panoramic radiographs and DUS images, thus determining the value of the panoramic radiographs' findings. PMID- 19903639 TI - Fatigue limits for composite restorations with and without glass ionomer cement liners. AB - This laboratory study compared the flexural endurance limits of clinical combinations of dental composite with and without glass ionomer cement (GIC) liners. Using only composite (Filtek Z350), specimens (10 mm long x 2 mm wide x 2 mm thick) in the control group were produced. Two GICs (Vitremer and Vitrebond) were used with the composite to prepare the test groups. Flexural strength and flexural fatigue limit (FFL) tests were performed. The FFL was determined using the staircase method. Data were analyzed by one-way ANOVA and Tukey's test. There was a significant difference in flexural strength values between the composite only specimens and those produced by composite and GIC (p < 0.05). No statistical difference was observed in the flexural strength values between composite with Vitremer and composite with Vitrebond (p < 0.05). No statistically significant differences were detected in FFL values between composite with Vitremer and composite with Vitrebond; in addition, the mean value of the composite-only specimens differed statistically from those of both composite with Vitremer and composite with Vitrebond (p < 0.05). The FFL was lower than the flexural strength, indicating a decrease in flexural strength of 45 to 50%. Using GICs with composite decreased the mechanical properties (FFL and flexural strength) of the composite. PMID- 19903640 TI - Comparative study of the effect of ultrasound on the removal of intracanal posts. AB - This study sought to evaluate how ultrasound affected the removal of stainless steel and titanium posts that had been attached with two different resin cements. The crowns of 32 maxillary canines were removed, the roots were embedded in acrylic resin blocks, and the canals were treated endodontically. The specimens were randomly distributed into two groups (n = 16) according to the brand of cement and subdivided (n = 8) according to the type of post. The specimens were submitted to ultrasonic vibration applied perpendicularly to the long axis of the tooth for 60 seconds. Data were submitted to ANOVA and showed no significant statistical difference among the groups (p > 0.05). It may be concluded that the effects of ultrasonic vibration used to remove intraradicular posts were not significantly different when applied to stainless steel or titanium posts cemented with chemically or dual-activated resin cements. PMID- 19903641 TI - Evaluation of desensitizing agents on dentin permeability and dentinal tubule occlusion: an in vitro study. AB - One hundred twelve specimens from bovine incisors were divided into eight groups: Group 1 (treated with 10% strontium chloride gel), Group 2 (treated with 2% sodium fluoride gel), Group 3 (treated with 2% stannous fluoride gel), Group 4 (treated with 5% potassium nitrate gel), Group 5 (treated with 10% potassium nitrate gel), Group 6 (treated with 3% potassium oxalate gel), Group 7 (treated with hydroxyethylcellulose gel), and Group 8 (which received no treatment). Dentinal tubules were exposed after 0.5 mm of deep abrasion using a carbide bur and EDTA gel application. After each treatment, dentin permeability, tubule occlusion, and chemical elements on dentin were analyzed. There was a significant difference among groups in dentin permeability (p < 0.05 ANOVA). Groups 4, 5, and 6 showed the lowest values, while Groups 1, 7, and 8 exhibited the highest. Groups 1, 2, 3, 7, and 8 showed open dentinal tubules, Groups 4 and 5 had partial tubule occlusion, and most of the tubules in Group 6 were obliterated. Energy dispersive x-rays revealed similar chemical characteristics among the experimental agents used, with traces of strontium, fluoride, sodium, and potassium. Within the limits of the study, 3% potassium oxalate gel showed the best results in terms of dentin permeability and dentinal tubule occlusion. PMID- 19903642 TI - Prevalence of oral lesions in the dental office. AB - Comprehensive head and neck examinations provide the best opportunity to carefully evaluate the oral cavity for the presence of disease. For this study, 40 dental professionals in 20 dental offices in Northern Ohio received a three hour standardization course on oral pathology, performed head and neck examinations on all adult patients over a three-week period, and recorded the presumptive identification of all observed oral lesions. Lesion-positive and lesion-negative patients were compared on patient age, gender, tobacco usage, denture status, and the type of provider (dentist or hygienist) making the observations. Providers performed head and neck examinations on 3,182 adults, 847 (26.7%) of whom had 1,073 lesions. Lesion-positive and negative patients were similar in terms of age and gender, although patients with lesions were almost twice as likely to use tobacco and more than 2.5 times as likely to wear dentures. Most lesions were classified as white (36.6%), followed by red (17%), ulcerated (14%), pigmented (14%), and soft tissue enlargements (12%). Leukoplakia and erythroplakia were observed in 1% of the screened adults, although patients were not aware of the presence of these conditions. Overall, the prevalence of lesions in dental patients (26.7%) was similar to national estimates (27.9%), but the prevalence of some specific lesions varied. PMID- 19903643 TI - Periodontal treatment with a low-level diode laser: clinical findings. AB - This study assessed the effects of low-level laser treatment in combination with scaling and root planing (SRP) in patients with periodontitis. Sixty subjects with chronic advanced periodontitis were assigned randomly to three treatment groups (n = 20) after collecting gingival clinical parameters. Group A received SRP on a single quadrant per day for four consecutive days; on the fifth day, all quadrants were rescaled. Group B received the same treatment as Group A, followed by laser application for five days. Group C received the same treatment as Group B but the laser treatment was administered for a total of 10 days. For Groups B and C, a low-level diode laser (630 to 670 nm) was used. The plaque index, gingival index, and sulcular bleeding index were recorded for all groups. For all clinical parameters, all three groups reported statistically significant differences (p < 0.005) compared to baseline data. Compared to Group A, Groups B and C showed statistically significant improvement for all clinical parameters. These findings suggest that a low-level diode laser can have a beneficial effect for treating inflammatory chronic advanced periodontitis. PMID- 19903644 TI - Vertical dimensional stability and rigidity of occlusal registration materials. AB - Dimensionally accurate occlusal registration records are essential for restorative dentistry; moreover, since records are not used immediately or may be used more than once, the registration material should exhibit accuracy over time (a concept known as dimensional stability). It has been speculated that materials with increased hardness or rigidity should produce more accurate registration records due to an increased resistance to distortion. This study compared the rigidity and associated dimensional accuracy of a recently marketed bisacrylic occlusal registration material and a vinyl polysiloxane (VPS). Maxillary and mandibular typodont arches were mounted on a plasterless articulator from which teeth No. 3, 13, and 15 had been removed to simulate edentulous spaces. After preparing teeth No. 2, 4, 12, and 14 as bridge abutments, the remaining teeth were equilibrated selectively to produce even anterior contact. Four digital photographs were taken to make vertical interarch measurements at four locations (teeth No. 3, 7, 10, and 14). Following initial photos (controls), 10 interocclusal records were made using each registration material, with material placed only in the segments in which teeth were prepared. The records were used for mounting the maxillary arch against the mandibular arch after 48, 72, and 120 hours. There were significant effects on vertical dimensional change related to arch location, material, and mounting time. Both materials demonstrated significantly larger posterior vertical openings than anterior vertical openings, while the bisacrylate produced a larger posterior opening than VPS at 48 and 72 hours and a larger anterior opening at all mounting times. There also was a significant difference in hardness/rigidity due to material and measurement time; at all measurement times, bisacrylate exhibited a significantly higher hardness number. PMID- 19903645 TI - Dental erosion due to excessive wine consumption. AB - Dental erosion can result in serious and irreversible damage to dentition, although it often is not recognized in the early stages. It is important to be aware of the numerous factors that can lead to tooth erosion, as this knowledge forms an important basis for intervention and treatment planning. This article presents a case of dental erosion involving an alcoholic woman who habitually consumed large quantities of white wine and discusses the dental erosion induced by excessive wine consumption. The authors also present the clinical aspects (including different etiologies of erosion) that distinguish extrinsic erosion from intrinsic erosion. PMID- 19903646 TI - Talon cusp: a case series. AB - Talon cusp is an uncommon dental developmental anomaly that typically appears as an accessory cusp-like structure projecting from the cingulum to the cutting edge in primary or permanent anterior teeth. This article presents 11 cases of talon cusp in seven patients. Three patients had unilateral talon cusps, one of which affected a permanent maxillary central incisor and included an unusual concurrent combination of dental anomalies; the other two affected permanent maxillary lateral incisors. The remaining eight talon cusps were observed bilaterally, with six affecting the permanent maxillary lateral incisors and the other two affecting the permanent maxillary canines. PMID- 19903647 TI - Maxillary radiolucency. Mesenchymal chondrosarcoma. PMID- 19903648 TI - Tender tooth. External resorption with granulation tissue. PMID- 19903649 TI - Costs and rehabilitation use of stroke survivors: a retrospective study of Medicare beneficiaries. AB - OBJECTIVE: To examine mortality, costs, and rehabilitation use in patients with stroke and stroke-related hemiparesis during a 4-year period following stroke onset. METHOD: This study was a retrospective, longitudinal claims analysis. Patients newly diagnosed with stroke and discharged from the hospital were identified from a 5% random sample of Medicare beneficiaries. Mortality, total Medicare costs, use of rehabilitation, and associated costs in stroke survivors with or without hemiparesis were the main outcome measures. RESULTS: Out of 4,604 newly diagnosed stroke patients, 1,166 developed hemiparesis. The 4-year mortality rate was significantly higher in the hemiparesis cohort than the nonhemiparesis cohort (55.2% vs. 47.5%; p < .01). The average Medicare cost per patient over the 4-year period was $77,143 for the hemiparesis cohort and $53,319 for the nonhemiparesis cohort (p < .01). A significantly higher proportion of patients in the hemiparesis cohort received rehabilitation than in the nonhemiparesis cohort (84% vs. 36% in Year 1, 30% vs. 10% in Year 2, 21% vs. 9% in Year 3, 16% vs. 7% in Year 4). Among patients who received rehabilitation, costs were significantly higher for the hemiparesis cohort ($17,680) than for the nonhemiparesis cohort ($7,841) in the fi rst year. While most rehabilitation costs for the hemiparesis cohort were incurred in the hospital inpatient setting in the fi rst year, the cost burden shifted to skilled nursing facilities and home health agencies in the following 3 years. CONCLUSIONS: Hemiparesis following stroke onset contributes to a higher mortality rate and higher Medicare costs in both the short and long term. PMID- 19903650 TI - The Three Cities Test: preliminary validation of a short bedside memory test in persons with acute stroke. AB - Screening tests for memory can be administered more quickly than standard tests of memory. They can be particularly useful with patients with acute medical illness or with the elderly who are unable to tolerate complex or lengthy memory testing, such as patients with acute stroke. However, screening measures for memory often lack validation and may have significant psychometric limitations. The purpose of this study was to validate and determine the psychometric properties of the Three Cities Test (TCT), a short test of memory that uses a selective reminding paradigm and the names of well-known cities as stimuli. The TCT was administered to 115 subjects: 60 patients with acute cerebrovascular accidents (Stroke group) and 55 age-matched orthopedic control patients (Ortho group). Results show that the TCT was significantly correlated with general measures of cognition (MMSE), another well-validated measure of learning and memory (HVLT-R), and clinical variables such as length of hospitalization and functional recovery. Compared to the Ortho group, the Stroke group had significantly worse performance on the TCT in terms of number of trials to criterion, delayed recall, and recognition discrimination. Preliminary results suggest that this instrument is well-received by patients with acute medical illness and cognitive impairment and that it possesses good construct and discriminative validity. Sensitivity and specificity performance as well as recommended cut scores are offered for the TCT. PMID- 19903651 TI - Rehabilitation of patients with chronic normal-pressure hydrocephalus after aneurysmal subarachnoid hemorrhage benefits from ventriculoperitoneal shunt. AB - This study compares the effect and rehabilitation prognosis of 6-month combined ventriculoperitoneal shunt (VPS) versus rehabilitation training alone in chronic normal-pressure hydrocephalus patients. METHOD: After suffering from aneurysmal subarachnoid hemorrhage, 39 subjects were diagnosed with chronic normal-pressure hydrocephalus because of clinical deterioration or nonimprovement of gait ataxia, cognitive disturbance, and/or urinary incontinence during rehabilitation. Twenty four subjects underwent ventriculoperitoneal shunt (VPS) placement, while 15 subjects did not undergo the procedure. RESULTS: Statistically significant increases in the Barthel Index (BI) were observed between the treatment and control groups 1 month (p < .05) and 6 months (p < .01) after VPS placement. Similar changes also were observed for the Mini-Mental State Examination (MMSE) at the 1-month (p < .01) and 6-month follow-up visits (p < .01). Significant differences in the change in MMSE and BI at admission, before VPS placement, and at 1 month and 6 months after VPS placement in the treatment group were demonstrated (p < .01). CONCLUSION: Chronic normal-pressure hydrocephalus after aneurysmal subarachnoid hemorrhage is an important issue to which clinicians should pay attention. It negatively influences the functional and behavioral outcome of rehabilitation but can be treated. PMID- 19903652 TI - Pediatric constraint-induced movement therapy: a promising intervention for childhood hemiparesis. AB - Experimental and clinical evidence is accumulating that supports the assertion that the damaged human brain is capable of responding to sensory stimulation in a sufficient manner to result in sustainable and useful benefits. The intensity and duration of therapeutic maneuvers that elicit improvement are under active investigation. Recent studies in animals, adults, and children with hemiparesis have shown that constraint of the less involved upper limb coupled with a behavioral program that repetitively encourages graded unilateral movements can result in long-term "new" functional activities. Constraint-induced movement therapy (CIMT) is a promising approach for treatment of children with stroke related hemiparesis from either prenatal or postnatal causes due to the enhanced neuroplasticity of the brain during early life. PMID- 19903653 TI - Understanding motor impairment in the paretic lower limb after a stroke: a review of the literature. AB - In addition to muscle weakness caused by injury to supraspinal centers, several mechanisms may contribute to motor impairment in the paretic lower limb following a stroke. Physiological changes in the paretic muscles and their motor units, passive or active restraint of agonist activation, and abnormal muscle activation patterns have been shown to occur after a stroke and to reduce muscle force generation. Other factors such as increased passive tone may impede agonist and antagonist muscle torque generation, while abnormal motor activation and altered motor control of muscles can produce abnormal gait patterns. Co-activation of opposing lower limb muscles contributes to joint stiffness and postural stability; abnormal co-activation in paretic lower limbs can lead to deficits in postural stabilization. Abnormal timing of muscle activation can also yield reduced muscle work output and, in turn, reduced limb function. When sensory deficits accompany muscle weakness, impaired processing of afferent signals may contribute to abnormal muscle activation, abnormal gait patterns, and abnormal responses to perturbation during gait and stance. This article reviews the impact of these various factors, individually and in combination, on impaired motor function in the paretic lower limb after a stroke. PMID- 19903655 TI - Gait changes following botulinum toxin A treatment in stroke. AB - PURPOSE: To characterize the effects of botulinum toxin A treatment of spastic plantar flexors in stroke on joint mobility and gait kinematics and kinetics. METHOD: Nine patients with hemiparetic stroke presenting with ankle hypertonicity participated in this exploratory open-label case series study. Comprehensive gait analysis provided bilateral kinematic and kinetic information for the ankle, knee, and hip joints throughout the stance phase. Data were obtained at baseline, 2 weeks, and 10 weeks post botulinum toxin injection of the spastic plantar flexors. RESULTS: Passive ankle range of motion increased post injection (p < .05). The amount of plantarflexion in late stance was significantly reduced (p < .05) while the maximum dorsiflexion increased in midstance at 10 weeks post treatment. The angular displacement profiles for the knee revealed that patients tended to display less hyperextension following treatment (p = .053). No significant changes in kinetic measures were found; however, case-by-case observations suggested that most patients experienced improvements in positive work production. CONCLUSIONS: The findings indicate that botulinum toxin treatment results in improved joint mobility and ankle kinematics and, in some patients, increases in positive work, suggesting better gait performance. PMID- 19903654 TI - Poststroke fear of falling in the hospital setting. AB - PURPOSE: Fear of falling (FoF) has a negative impact on older adults, however there is a paucity of research regarding the development and impact of FoF after stroke. Therefore, our objectives were to determine the proportion of individuals with FoF and the affect of FoF during the immediate poststroke period. METHODS: This observational study of baseline data from a pilot cohort study includes a convenience sample of 28 adults with acute stroke before discharge home. Measures include self-reported FoF, the Falls Efficacy Scale-Swedish Version [FES(S)], Stroke-Specific Quality of Life (SS-QOL), performance and satisfaction with performance (Canadian Occupational Performance Measure), anxiety (Generalized Anxiety Disorder-7), and depression (Patient Health Questionnaire-9). RESULTS: Fifteen (54%) of the participants reported FoF. Those with FoF were more likely to have decreased SS-QOL domain scores for energy (p = .013), personality (p = .015), and thinking (p = .008); decreased performance (self-care, productivity, and leisure) (p = .019) and satisfaction with performance (p = .010); and increased anxiety (p = .002) than those without FoF. CONCLUSIONS: Those with FoF demonstrated significantly increased anxiety and showed decreased performance and satisfaction with performance, energy, thinking, and personality than those without FoF. This suggests that poststroke FoF is related not only to physical challenges but also to cognitive and emotional factors in the poststroke period. Identifying and treating these conditions should be evaluated as a means to decrease FoF and improve outcomes post stroke. PMID- 19903656 TI - Stroke education: promising effects on the health beliefs of those at risk. AB - BACKGROUND: The effect of patient education on reducing stroke has had mixed effects, raising questions about how to achieve optimal benefit. Because past evaluations have typically lacked an appropriate theoretical base, the design of past research may have missed important effects. METHOD: This study used a social cognitive framework to identify variables that might change in response to education. A mixed design was used to evaluate two approaches to an intervention, both of which included education. Fifty seniors completed a measure of stroke knowledge and beliefs twice: before and after an intervention that was either standard (educational brochure plus activities that were not about stroke) or enhanced (educational brochure plus activities designed to enhance beliefs about stroke). Outcome measures were health beliefs, intention to exercise to reduce stroke, and stroke knowledge. RESULTS: Selected beliefs changed significantly over time but not differentially across conditions. Beliefs that changed were (a) perceived susceptibility to stroke and (b) perceived benefit of exercise to reduce risk. Benefit beliefs, in particular, were strongly and positively associated with intention to exercise. CONCLUSION: Findings suggest that basic approaches to patient education may influence health beliefs. More effective stroke prevention programs may result from continued consideration of the role of health beliefs in such programs. PMID- 19903657 TI - Inferring the evolutionary history of gene clusters from phylogenetic and gene order data. AB - Gene duplication is frequent within gene clusters and plays a fundamental role in evolution by providing a source of new genetic material upon which natural selection can act. Although classical phylogenetic inference methods provide some insight into the evolutionary history of a gene cluster, they are not sufficient alone to differentiate single- from multiple gene duplication events and to answer other questions regarding the nature and size of evolutionary events. In this paper, we present an algorithm allowing to infer a set of optimal evolutionary histories for a gene cluster in a single species, according to a general cost model involving variable length duplications (in tandem or inverted), deletions, and inversions. We applied our algorithm to the human olfactory receptor and protocadherin gene clusters, showing that the duplication size distribution differs significantly between the two gene families. The algorithm is available through a web interface at http://www lbit.iro.umontreal.ca/DILTAG/. PMID- 19903658 TI - Bioactive rather than total IGF-I is involved in acute responses to nutritional interventions in CAPD patients. AB - BACKGROUND: Inadequate food intake plays an important role in the development of malnutrition in continuous ambulatory peritoneal dialysis (CAPD) patients. Aim of the study. The aim of the study was to investigate in CAPD patients whether circulating insulin-like growth factor-I (IGF-I) bioactivity may offer a more sensitive index to acute nutritional interventions than total IGF-I. METHODS: An open-label, randomized, crossover study of 2 days-with a 1-week interval-was performed in 12 CAPD patients in the fed state to compare a mixture of amino acids (Nutrineal 1.1%) plus glucose (AA plus G) (Physioneal 1.36% to 3.86%) dialysate versus G only as control dialysate. Fed-state conditions were created by identical liquid hourly meals. IGF-I bioactivity was measured by the kinase receptor activation assay (IGF-I KIRA); total IGF-I was measured by immunoassay. RESULTS: In the fed state, both after AA plus G as well as after G dialysis IGF-I bioactivity increased compared to baseline, while no changes in circulating total IGF-I levels were observed in both treatment arms. However, the increase in IGF-I bioactivity was only significant after AA plus G dialysis (P = 0.02). CONCLUSIONS: Our results provide evidence that in CAPD patients changes in circulating IGF-I bioactivity are associated with nutrient intake and that IGF-I bioactivity rather than total IGF-I is involved in acute responses to nutritional interventions in CAPD patients. PMID- 19903659 TI - Effects of end-stage renal disease and haemodialysis on dendritic cell subsets and basal and LPS-stimulated cytokine production. AB - BACKGROUND: Although bacterial infections have dramatically declined as a cause of death in the general population, they remain a major cause of mortality in patients with end-stage renal disease (ESRD). Moreover, the response to vaccination is profoundly impaired in this population. Dendritic cells (DC) are the major antigen-presenting cells that bridge the innate and adaptive immune responses. Activation of DC by pathogens results in secretion of inflammatory cytokines and up-regulation of co-stimulatory molecules. The activated DC prime naive T and B cells to the captured antigens. METHODS: Using flow cytometry, the number and phenotype of circulating DC [myeloid DC (mDC) and plasmacytoid DC (pDC)] were quantified in pre- and post-dialysis blood samples from 20 ESRD patients maintained on haemodialysis. Ten normal individuals served as controls. In addition, the level of DC activation and their response to lipopolysaccharide (LPS) stimulation were determined by assessing expression of co-stimulatory molecule, CD86, and antigen-presenting molecule, HLA-DR, as well as production of TNFalpha, IFNalpha and IL-6. RESULTS: Compared to the control group, the circulating dendritic cell count was significantly reduced in the ESRD patients before dialysis and declined further after dialysis. The reduction in pDC numbers was more striking than mDC. The magnitude of the LPS-induced up-regulation of CD86 was comparable among the study groups as well as pre- and post-dialysis samples. However, LPS-induced TNFalpha production was significantly reduced in the post-dialysis samples with no significant difference in IL-6 and IFNalpha productions among the study groups and in pre- and post-dialysis samples. CONCLUSIONS: ESRD results in significant DC depletion which is largely due to diminished plasmacytoid DC subset. Haemodialysis procedure intensifies DC depletion and impairs LPS-induced TNFalpha production. PMID- 19903660 TI - Impact of renin-angiotensin-aldosterone blockade by angiotensin-converting enzyme inhibitors or AT-1 blockers on frequency of contrast medium-induced nephropathy: a post-hoc analysis from the Dialysis-versus-Diuresis (DVD) trial. AB - BACKGROUND: After exposure to contrast medium (CM), about 10% of patients will develop contrast medium-induced nephropathy (CIN), with severe consequences for their prognosis. Although numerous studies evaluated risk factors for CIN development, it is still a matter of debate whether treatment with angiotensin converting enzyme inhibitors (ACE-I) or AT-1 blockers increases the frequency of CIN after exposure to CM or not. METHODS: We performed a prospective, single centre study (January 2001-July 2004) to compare different treatments for CIN prevention. Creatinine levels within 72 h after CM application and in-hospital outcomes were documented. The impact of RAAS blockade on the frequency of CIN was assessed retrospectively. RESULTS: Four hundred twelve patients were included (83.5% men, 29.1% diabetes mellitus, 74.6% hypertension). Of these, 269 patients (65.3%) were taking ACE-I (n = 236) or AT-1 blockers (n = 33). There were no significant differences in mean age (P = 0.075), creatinine levels (P = 0.113), gender (P = 0.281), diabetes mellitus (P = 0.172) or left ventricular ejection fraction (P = 0.09) between patients treated or not treated with RAAS blockade. Univariate analyses concerning development of CIN depending on treatment with RAAS blockade within 72 h found CIN to be significantly higher in patients treated with RAAS blockade (11.9 vs 4.2%, P = 0.006). Multivariate analyses (logistic regression) identified RAAS blockade to be an independent predictor of CIN (odds ratio 3.082, 95% confidence interval 1.234-7.698, P = 0.016). CONCLUSION: Patients treated with RAAS blockade before exposure to CM develop significantly more often CIN within 72 h. Even after adjustment for confounding comorbidities, treatment with ACE-I or AT-1 blockers turned out to be an independent risk predictor. PMID- 19903661 TI - Feasibility of combined treatment with enalapril and candesartan in advanced chronic kidney disease. AB - BACKGROUND: Dual blockade of the renin-angiotensin system (RAS) has been claimed to have a specific renal protective effect in chronic kidney disease (CKD). The present short-term study reports on the feasibility of dual blockade in a consecutive group of patients with CKD stage 3-5. METHODS: Forty-seven CKD patients, mean age 59 years, with mean estimated glomerular filtration rate (GFR) 26 ml/min/1.73 m(2) (range 13-49) and blood pressure (BP) 133/78 mmHg, were block randomized in an open study to 16 weeks of monotherapy with increasing doses of RAS blockade aiming at enalapril 20 mg o.d. or candesartan 16 mg o.d. Thereafter, the complementary drug was added in incremental doses over a period of 5 weeks aiming at combined enalapril 20 mg and candesartan 16 mg for 3 weeks. Seventy five percent of the patients were known to be RAS blockade tolerant. Blood samples and BP were measured every 2-3 weeks. Doses of study medication were reduced in case of hyperkalemia >5.5 mmol/l, a sustained rise in p-creatinine >30% or symptomatic hypotension. RESULTS: Twenty-one patients (45%) did not tolerate dual blockade in aimed dosages due to unacceptable p-creatinine increase (n = 12, including two study withdrawals), hypotension (n = 6), general discomfort (n = 2) or unmanageable hyperkalemia (n = 1). Hyperkalemia >5.5 mmol/l was seen in seven patients (15%). The reduced-dose group had baseline lower eGFR and diastolic BP. CONCLUSIONS: Forty-five percent of CKD stage 3-5 patients did not tolerate dual RAS blockade with 20 mg enalapril and 16 mg candesartan daily, primarily due to loss of renal function or hypotension. Hyperkalemia could be managed in most patients. Caution is recommended when giving this treatment to patients with advanced CKD. PMID- 19903662 TI - Rapamycin, unlike cyclosporine A, enhances suppressive functions of in vitro induced CD4+CD25+ Tregs. AB - BACKGROUND: A growing body of data shows that CD4(+)CD25(+) regulatory T cells (Tregs) can induce transplantation tolerance by suppressing immune responses to allograft antigens. However, both the generation and the suppressive capacity of CD4(+)CD25(+) Tregs can be substantially affected by different immunosuppressive drugs used in clinical transplantation. The goal of this study was to compare the effects of cyclosporine A and rapamycin on the induction and suppressive functions of human CD4(+)CD25(+) Tregs in vitro. METHODS: CD4(+)CD25(+) Tregs were induced in two-way mixed lymphocyte reaction (MLR) in the presence of rapamycin (Treg-Rapa) or cyclosporine A (Treg-CsA). Tregs were identified in MLR cultures by flow cytometry using anti-CD4, anti-CD25, anti-CTLA-4, anti-CD122, anti-GITR mAbs and ant-PE-FOXP3 staining sets. Suppressive capacity of induced Tregs was evaluated by their capability to inhibit anti-CD3 Ab-triggered proliferation of peripheral blood mononuclear cells (PBMCs), as measured by flow cytometry. The concentration of TGF-beta1 in culture supernatants was measured by enzyme-linked immunosorbent assay. RESULTS: Although both rapamycin and cyclosporine A suppressed the induction of CD4(+)CD25(+) Tregs during MLRs, this effect was significantly more pronounced in cells cultured with cyclosporine. On the other hand, only rapamycin significantly decreased the percentage of CD4(+)CD25(+) Tregs which expressed GITR, a negative regulator of Treg's suppressive capacity. Importantly, Treg-Rapa, unlike Treg-CsA, displayed significant suppressive activity and were capable of inhibiting the proliferation of anti-CD3 Ab-activated PBMCs. This activity was likely mediated by TGF-beta1. CONCLUSIONS: Rapamycin, unlike cyclosporine A, does not inhibit the function of CD4(+)CD25(+) Tregs. This implies that rapamycin could contribute to the development of transplantation tolerance by promoting the induction of functional CD4(+)CD25(+) Tregs. Moreover, our results suggest that rapamycin could be combined with functional Tregs. PMID- 19903667 TI - A very prolonged asystolic vasovagal syncope is suspended but not aborted by counterpressure manoeuvre. PMID- 19903668 TI - Intrinsic neural reflexes in the post-transplant human heart. AB - Radiofrequency ablation of post-transplant flutter in a centrally denervated donor atrium at a site remote from the AV node resulted in transient worsening of AV nodal conduction, with absent central vagal reinnervation. This could be an electrophysiological marker of intact innervation to the donor AV node from the intrinsic cardiac neuronal plexus, not demonstrated in human hearts earlier. PMID- 19903669 TI - A new approach to confirming or excluding ventricular pre-excitation on a 12-lead ECG. AB - AIMS: The purpose of this study was to determine simple features of the standard 12-lead electrocardiogram (ECG) and incorporate them in a stepwise algorithm that would help confirm or exclude the presence of ventricular pre-excitation. METHODS AND RESULTS: We retrospectively analysed multiple variables on pre- and post ablation ECGs in 238 patients with manifest accessory pathways that had been successfully ablated. A new variable, PR dispersion, was defined as a difference between maximum and minimum PR intervals on a single 12-lead ECG. A logistic regression analysis showed the combination of the following criteria to be powerful in the confirmation of the diagnosis in patients with suspected delta wave: presence of both PR interval < or = 120 ms and PR dispersion > or = 20 ms, absence of initial positive deflection (septal R wave) in lead augmented voltage right arm (aVR), and horizontal QRS transition in lead V1 or before. A stepwise algorithm was developed based on these criteria. Of the total 476 ECGs, seven patients with pre-excitation and one patient with normal ECG were misdiagnosed using the algorithm. Even though the retrospectively determined sensitivity and specificity of the three stepwise criteria were high (97% and 99%, respectively) a prospective study evaluating the algorithm is needed. CONCLUSION: Using a stepwise approach is a very sensitive and specific technique for excluding or confirming ventricular pre-excitation on a 12-lead ECG. PMID- 19903670 TI - Chronic total occlusion of left circumflex artery after radiofrequency ablation of left ventricular outflow tract tachycardia. AB - In this report, we present a 22-year-old female patient referred to our institution for evaluation of anginal chest pain. Her medical history revealed two ablation procedures of the left ventricular outflow tract tachycardia performed 1 month a part, 2 years ago. Coronary angiography revealed chronic total occlusion of the proximal left circumflex artery. To our knowledge, this is the first report of ablation-related chronic total occlusion of a coronary artery. PMID- 19903671 TI - Trend of the main clinical characteristics and pacing modality in patients treated by pacemaker: data from the Italian Pacemaker Registry for the quinquennium 2003-07. AB - AIMS: To assess the impact on clinical practice of the major published studies, we report the information collected by the Italian Pacemaker Registry (IPR) in the quinquennium 2003-07. METHODS AND RESULTS: The IPR collected prospectively main epidemiological, clinical, and electrocardiographic data of patients treated by pacemaker (PM) in Italy on the basis of European PM Card. The number of reported PMs in Italy was 30,820 in 2003, 32,047 in 2004, 31,870 in 2005, 31,813 in 2006, and 31,146 in 2007, respectively. The median age was 79 years in all 5 years. Among the atrio-ventricular (AV) conduction defects, third-degree AV block was the most common occurrence. Of the sick sinus syndrome (SSS), sinus node dysfunction involved the majority of cases followed by bradycardia-tachycardia syndrome. Year-over-year percentages among the different indications remained stable. Syncope and dizzy spells were by far the most common symptoms. Dual chamber pacing showed an increasing utilization in all the examined years. CONCLUSION: Italian PM Registry data for the study period reveal a stable pattern of PM utilization and indications. A higher use of dual-chamber pacing in comparison to single-chamber pacing was reported for all indications, despite inconclusive data of the major randomized trials. PMID- 19903672 TI - Which is more useful nomogram or equation? PMID- 19903673 TI - The utility of a prognostic index for predicting time to first treatment in early chronic lymphocytic leukemia: the GIMEMA experience. AB - BACKGROUND: A prognostic index based on widely available clinical and laboratory features was recently proposed to predict survival in patients with previously untreated chronic lymphocytic leukemia. We assessed the utility of this index for predicting time to first treatment in early chronic lymphocytic leukemia. DESIGN AND METHODS: An observational database of the GIMEMA (Gruppo Italiano Malattie EMatologiche dell'Adulto), which included 310 patients with newly diagnosed Binet stage A chronic lymphocytic leukemia who were observed at different primary hematology centers during the period 1991 - 2000, was used for the purpose of this study. RESULTS: The new prognostic index enabled Binet stage A patients to be divided into two subgroups that differed with respect to time to first treatment (P=0.003). The original prognostic index was derived from a database that included cases observed at a reference academic center; these patients were younger (P<0.0001) and had more advanced disease (P<0.0001) than those in the current investigation, which studied community-based patients whose data were recorded at presentation. With this in mind, we used an optimal cut-off search to determine how best to split patients with Binet stage A disease into different prognostic groups. According to the recursive partitioning (RPART) model, a classification tree was built that identified three subsets of patients who scores were 0-2 (low risk), 3-4 (intermediate risk) and 5-7 (high risk). The probability of remaining free from therapy at 5 years was 100% in the low risk group, 81.2% in the intermediate risk group and 61.3% in the high risk group (P<0.0001). CONCLUSIONS: The results of this study confirm the utility of a new prognostic index for predicting time to first treatment in a large sample series of community-based patients with early stage chronic lymphocytic leukemia at presentation. Our effort to develop a revised scoring method meets the need to separate Binet stage A patients into different prognostic groups in order to devise individualized and tailored follow-up during the treatment-free period. PMID- 19903674 TI - Additional chromosome abnormalities in patients with acute promyelocytic leukemia treated with all-trans retinoic acid and chemotherapy. AB - BACKGROUND: Acute promyelocytic leukemia is a subtype of acute myeloid leukemia characterized by the t(15;17). The incidence and prognostic significance of additional chromosomal abnormalities in acute promyelocytic leukemia is still a controversial matter. DESIGN AND METHODS: Based on cytogenetic data available for 495 patients with acute promyelocytic leukemia enrolled in two consecutive PETHEMA trials (LPA96 and LPA99), we analyzed the incidence, characteristics, and outcome of patients with acute promyelocytic leukemia with and without additional chromosomal abnormalities who had been treated with all-trans retinoic acid plus anthracycline monochemotherapy for induction and consolidation. RESULTS: Additional chromosomal abnormalities were observed in 140 patients (28%). Trisomy 8 was the most frequent abnormality (36%), followed by abn(7q) (5%). Patients with additional chromosomal abnormalities more frequently had coagulopathy (P=0.03), lower platelet counts (P=0.02), and higher relapse-risk scores (P=0.02) than their counterparts without additional abnormalities. No significant association with FLT3/ITD or other clinicopathological characteristics was demonstrated. Patients with and without additional chromosomal abnormalities had similar complete remission rates (90% and 91%, respectively). Univariate analysis showed that additional chromosomal abnormalities were associated with a lower relapse-free survival in the LPA99 trial (P=0.04), but not in the LPA96 trial. However, neither additional chromosomal abnormalities overall nor any specific abnormality was identified as an independent risk factor for relapse in multivariate analysis. CONCLUSIONS: The lack of independent prognostic value of additional chromosomal abnormalities in acute promyelocytic leukemia does not support the use of alternative therapeutic strategies when such abnormalities are found. PMID- 19903675 TI - Class II-associated invariant chain peptide down-modulation enhances the immunogenicity of myeloid leukemic blasts resulting in increased CD4+ T-cell responses. AB - BACKGROUND: Disease recurrence in patients with acute myeloid leukemia may be partially explained by the escape of leukemic blasts from CD4(+) T-cell recognition. The current study investigates the role of aberrant HLA class II antigen presentation on leukemic blasts by determining both the clinical and functional impact of the class II-associated invariant chain peptide (CLIP). DESIGN AND METHODS: The levels of expression of CLIP and HLA-DR on blood and bone marrow samples from 207 patients with acute myeloid leukemia were correlated with clinical outcome. Irradiated CLIP(-) and CLIP(+) leukemic blasts were compared for their ability to induce CD4(+) T cells during mixed leukocyte reactions. To discriminate between these blasts, we down-modulated CLIP expression on myeloid leukemic cell lines by RNA interference of the invariant chain, a chaperone protein critically involved in HLA-DR processing, and performed flow cytometric sorting for their isolation from primary acute myeloid leukemia samples. RESULTS: We found that patients with leukemic blasts characterized by a high amount of HLA DR occupied by CLIP (relative amount of CLIP) had a significantly shortened disease-free survival. The clear reductions in amount of HLA-DR occupied by CLIP on blasts of the THP-1 and Kasumi-1 myeloid leukemic cell lines after treatment with invariant chain short interfering RNA resulted in enhanced rates of allogeneic CD4(+) T-cell proliferation. Similar findings were obtained in an autologous setting, in which there were strong increases in proliferation of remission CD4(+) T cells stimulated with CLIP(-)-sorted leukemic blasts from HLA DR(+) acute myeloid leukemia patients, in contrast to CLIP(+)-sorted leukemic blasts from the same patients. CONCLUSIONS: These data highlight the relevance of CLIP expression on leukemic blasts and the potential of CLIP as a target for immunomodulatory strategies to enhance HLA class II antigen presentation and CD4(+) T-cell reactivity in acute myeloid leukemia. PMID- 19903676 TI - Pregnancy and beta-thalassemia: an Italian multicenter experience. AB - BACKGROUND: Recent advances in the management of thalassemia have significantly improved life expectancy and quality of life of patients with this hemoglobinopathy, with a consequent increase in their reproductive potential and desire to have children. DESIGN AND METHODS: We describe the methods of conception and delivery, as well as the course and outcome of pregnancy including transfusions, iron overload and chelation in 46 women with thalassemia major (58 pregnancies) and in 11 women with thalassemia intermedia (17 pregnancies). Conception was achieved after gonadotrophin-induced ovulation in 33 of the women with thalassemia major and spontaneously in all of those with thalassemia intermedia. RESULTS: Among the women with thalassemia major, 91% of the pregnancies resulted in successful delivery of 45 singleton live-born neonates, five sets of twins and one set of triplets. No secondary complications of iron overload developed or worsened during pregnancy. When considering only the singleton pregnancies, the proportion of babies with intrauterine growth retardation did not differ from that reported in the general Italian population. The high prevalence of pre-term births (32.7%) was mostly related to multiple pregnancies and precautionary reasons. Pregnancy was safe in most women with thalassemia major or intermedia. However, women with thalassemia intermedia who had never previously been transfused or who had received only minimal transfusion therapy were at risk of severe alloimmune anemia if blood transfusions were required during pregnancy. CONCLUSIONS: Provided that a multidisciplinary team is available, pregnancy is possible, safe and usually has a favorable outcome in patients with thalassemia. In women with hypogonadotropic hypogonadism, gonadal function is usually intact and fertility is usually retrievable. PMID- 19903677 TI - IgA and IgG hypogammaglobulinemia in Waldenstrom's macroglobulinemia. AB - BACKGROUND: Hypogammaglobulinemia is common in Waldenstrom's macroglobulinemia. The etiology of this finding remains unclear, but it has been speculated to be based on tumor-induced suppression of the 'uninvolved' immunoglobulin production DESIGN AND METHODS: We evaluated the incidence of IgA and IgG hypogammaglobulinemia in 207 untreated patients with Waldenstrom's macroglobulinemia and investigated the associated clinicopathological findings and impact of therapy. We also sequenced eight genes (AICDA, BTK, CD40, CD154, NEMO, TACI, SH2D1A, UNG) implicated in immunoglobulin deficiency in 19 Waldenstrom's macroglobulinemia patients with IgA and/or IgG hypogammaglobulinemia. RESULTS: At baseline 63.3%, 58.0% and 49.3% of the 207 patients had abnormally low serum levels of IgA, IgG, or both. No association between IgA and IgG hypogammaglobulinemia and disease burden, serum IgM levels, beta(2)-microglobulin, International Prognostic Scoring System score, or incidence of recurrent infections was observed, although the presence of adenopathy and/or splenomegaly was associated with a lower incidence of hypogammaglobulinemia. Lower IgA and IgG levels were associated with disease progression in patients managed with a 'watch and wait' strategy. IgA and/or IgG levels remained abnormally low despite response to treatment, including complete remissions. A missense mutation in the highly conserved catalytic site of UNG was observed in a patient with hypogammaglobulinemia, warranting further study of this pathway in Waldenstrom's macroglobulinemia. CONCLUSIONS: IgA and IgG hypogammaglobulinemia is common in Waldenstrom's macroglobulinemia and persists despite therapeutic intervention and response. IgA and IgG hypogammaglobulinemia does not predict the risk of recurrent infections in patients with Waldenstrom's macroglobulinemia, although lower levels of serum IgA and IgG are associated with disease progression in Waldenstrom's macroglobulinemia patients being managed with a 'watch and wait' strategy. PMID- 19903678 TI - Prognostic impact of pre-transplantation transfusion history and secondary iron overload in patients with myelodysplastic syndrome undergoing allogeneic stem cell transplantation: a GITMO study. AB - BACKGROUND: Transfusion-dependency affects the natural history of myelodysplastic syndromes. Secondary iron overload may concur to this effect. The relative impact of these factors on the outcome of patients with myelodysplastic syndrome receiving allogeneic stem-cell transplantation remains to be clarified. DESIGN AND METHODS: We retrospectively evaluated the prognostic effect of transfusion history and iron overload on the post-transplantation outcome of 357 patients with myelodysplastic syndrome reported to the Gruppo Italiano Trapianto di Midollo Osseo (GITMO) registry between 1997 and 2007. RESULTS: Transfusion dependency was independently associated with reduced overall survival (hazard ratio=1.48, P=0.017) and increased non-relapse mortality (hazard ratio=1.68, P=0.024). The impact of transfusion-dependency was noted only in patients receiving myeloablative conditioning (overall survival: hazard ratio=1.76, P=0.003; non-relapse mortality: hazard ratio=1.70, P=0.02). There was an inverse relationship between transfusion burden and overall survival after transplantation (P=0.022); the outcome was significantly worse in subjects receiving more than 20 red cell units. In multivariate analysis, transfusion dependency was found to be a risk factor for acute graft-versus-host disease (P=0.04). Among transfusion-dependent patients undergoing myeloablative allogeneic stem cell transplantation, pre-transplantation serum ferritin level had a significant effect on overall survival (P=0.01) and non-relapse mortality (P=0.03). This effect was maintained after adjusting for transfusion burden and duration, suggesting that the negative effect of transfusion history on outcome might be determined at least in part by iron overload. CONCLUSIONS: Pre transplantation transfusion history and serum ferritin have significant prognostic value in patients with myelodysplastic syndrome undergoing myeloablative allogeneic stem cell transplantation, inducing a significant increase of non-relapse mortality. These results indicate that transfusion history should be considered in transplantation decision-making in patients with myelodysplastic syndrome. PMID- 19903679 TI - Mutations of JAK2 and TET2, but not CBL are detectable in a high portion of patients with refractory anemia with ring sideroblasts and thrombocytosis. PMID- 19903680 TI - Successful eradication of acquired factor-VIII-inhibitor using single low-dose rituximab. PMID- 19903681 TI - Amyloidosis relapsing after autologous stem cell transplantation treated with bortezomib: normalization of detectable serum-free light chains and reversal of tissue damage with improved suitability for transplant. PMID- 19903682 TI - Chronic nitrate therapy is associated with different presentation and evolution of acute coronary syndromes: insights from 52,693 patients in the Global Registry of Acute Coronary Events. AB - AIMS: Brief episode(s) of ischaemia may increase cardiac tolerance to a subsequent major ischaemic insult ('preconditioning'). Nitrates can pharmacologically mimic ischaemic preconditioning in animals. In this study, we investigated whether antecedent nitrate therapy affords protection toward acute ischaemic events using data from the Global Registry of Acute Coronary Events. METHODS AND RESULTS: The dataset comprised 52,693 patients from 123 centres in 14 countries: 42,138 (80%) were nitrate-naive and 10,555 (20%) were on chronic nitrates at admission. In nitrate-naive patients, admission diagnosis was ST segment elevation myocardial infarction (STEMI) in 41%, whereas 59% presented with non-ST-segment elevation acute coronary syndrome (NSTE-ACS). In contrast, only 18% nitrate users showed STEMI, whereas 82% presented with NSTE-ACS. Thus, among nitrate users clinical presentation was tilted toward NSTE-ACS by more than four-fold, STEMI occurring in less than one of five patients (P < 0.0001). After adjustment (age, sex, medical history, prior therapy, revascularization, previous angina), chronic nitrate use remained independent predictor of NSTE-ACS (OR 1.36; 95% CI 1.26-1.46; P < 0.0001). Furthermore, regardless of presentation, within both STEMI and NSTEMI populations, antecedent nitrate use was associated with significantly lower levels of CK-MB and troponin (P < 0.0001 for all). CONCLUSION: In this large multinational registry, chronic nitrate use was associated with a shift away from STEMI in favour of NSTE-ACS and with less release of markers of cardiac necrosis. These findings suggest that in nitrate users acute coronary events may develop to a smaller extent. Randomized, placebo controlled trials are warranted to establish whether nitrate therapy may pharmacologically precondition the heart toward ischaemic episodes. PMID- 19903683 TI - Continuous positive airway pressure increases haemoglobin O2 saturation after acute but not prolonged altitude exposure. AB - AIMS: It is unknown whether subclinical high-altitude pulmonary oedema reduces spontaneously after prolonged altitude exposure. Continuous positive airway pressure (CPAP) removes extravascular lung fluids and improves haemoglobin oxygen saturation in acute cardiogenic oedema. We evaluated the presence of pulmonary extravascular fluid increase by assessing CPAP effects on haemoglobin oxygen saturation under acute and prolonged altitude exposure. METHODS AND RESULTS: We applied 7 cm H(2)O CPAP for 30 min to healthy individuals after acute (Capanna Margherita, CM, 4559 m, 2 days permanence, and <36 h hike) and prolonged altitude exposure (Mount Everest South Base Camp, MEBC, 5350 m, 10 days permanence, and 9 days hike). At CM, CPAP reduced heart rate and systolic pulmonary artery pressure while haemoglobin oxygen saturation increased from 80% (median), 78-81 (first to third quartiles), to 91%, 84-97 (P < 0.001). After 10 days at MEBC, haemoglobin oxygen saturation spontaneously increased from 77% (74-82) to 86% (82-89) (P < 0.001) while heart rate (from 79, 64-92, to 70, 54-81; P < 0.001) and respiratory rate (from 15, 13-17, to 13, 13-15; P < 0.001) decreased. Under such conditions, these parameters were not influenced by CPAP. CONCLUSION: After ascent excessive lung fluids accumulate affecting haemoglobin oxygen saturation and, in these circumstances, CPAP is effective. Acclimatization implies spontaneous haemoglobin oxygen saturation increase and, after prolonged altitude exposure, CPAP is not associated with HbO(2)-sat increase suggesting a reduction in alveolar fluids. PMID- 19903685 TI - Throw-off instruments for advanced thoracoscopic procedures. AB - Performing complex thoracoscopic procedures can necessitate the use of multiple instruments and, consequently, the use of multiple ports. This results in parietal harm and in overcrowding of the operative field with instrument conflicts. We present the interest of using lung retractors and vascular clamps that can be released inside the chest cavity without blocking a trocar access. PMID- 19903684 TI - Safety and efficacy of drug-eluting vs. bare metal stents in patients with diabetes mellitus: long-term follow-up in the Swedish Coronary Angiography and Angioplasty Registry (SCAAR). AB - AIMS: Patients with diabetes mellitus have more extensive coronary artery disease, more disease progression, and restenosis. The use of drug-eluting stents (DES) in these patients is widespread, despite uncertain long-term safety and efficacy. METHODS AND RESULTS: All consecutive patients with diabetes mellitus in Sweden who underwent percutaneous coronary intervention were entered into the Swedish Coronary Angiography and Angioplasty Registry (SCAAR) during 2003-06 with complete follow-up for 1-4 years (median 2.5). Patients who received at least one DES (n = 4754) were compared with those who received only bare metal stents (BMS) (n = 4956) at the index procedure. Combined outcome of death or myocardial infarction (MI) showed no difference for DES vs. BMS, relative risk (RR), 0.91 [95% confidence interval (CI), 0.77-1.06]. Myocardial infarction was significantly less common with DES in patients who received only one stent RR, 0.80 (95% CI, 0.66-0.96). The restenosis rate was 50% lower in DES-treated patients RR, 0.50 (95% CI, 0.35-0.70) and was associated with a higher adjusted RR of MI, RR, 5.03 (95% CI, 4.25-5.97). DES was associated with reduced restenosis rates in all subgroups of diabetic patients with the greatest benefit in stent diameters <3 mm or stent length >20 mm. The number of lesions treated with DES to prevent one restenosis ranged from 11 to 47 in various subgroups. CONCLUSION: This real-life registry study shows that restenosis was halved by DES in diabetic patients with stable or unstable coronary disease, with similar risk of death or MI up to 4 years compared with BMS. PMID- 19903686 TI - Does a skeletonized or pedicled right gastro-epiploic artery improve patency when used as a conduit in coronary artery bypass graft surgery? AB - A best evidence topic in cardiac surgery was written according to a structured protocol. The question addressed was whether skeletonization of the right gastro epiploic artery (RGEA) improves graft patency in coronary artery bypass grafting (CABG). Altogether >25 papers were found using the reported search, of which 11 papers represented the best evidence to answer this clinical question. The author, journal, date and country of publication, patient group studied, study type, relevant outcomes and results are tabulated. Four out of the 11 papers were comparative studies (skeletonized conduits vs. pedicled conduits) and four studies produced one-year follow-up data. No studies revealed long-term patency rates as there was no follow-up data beyond five years. It is important to note that the evidence in the literature is based in a Japanese population. The vast majority of the target vessel which had been grafted by the RGEA was the right coronary artery and more specifically the posterior descending artery (PDA). The association between off-pump technique, sequential grafting, skeletonization of the RGEA with the harmonic scalpel and angiographic patency has not been adequately assessed. The studies reveal excellent patency rates for both early and mid-term skeletonized RGEA conduits. Overall patency rates were 97.7% within three months, 92.4% at a mean of approximately 1 year, 91.5% at a mean of approximately 2 years, and 86.4% at 4 years. In the four comparative studies, skeletonization patency was at least comparable and in one study superior to pedicled conduits. One study revealed a higher four-year cumulative patency rate for skeletonized conduits in comparison to a previous study by the same author where pedicled grafts were used. In conclusion, patency rates exceeded 95% in 10 studies for a follow-up of up to three months postoperatively. The evidence which supports the use of a 'skeletonized' RGEA is growing and this paper demonstrates clearly that in terms of patency, a skeletonized RGEA to the PDA should be considered as a conduit for CABG surgery especially when total arterial revascularization strategy with in situ conduits and no manipulation of the ascending aorta is the treatment of choice. PMID- 19903687 TI - Ultrasound estimation of volume of postoperative pleural effusion in cardiac surgery patients. AB - The aim of this study was to establish a practical simplified formula to facilitate the management of a frequently occurring postoperative complication, pleural effusion. Chest ultrasonography with better sensitivity and reliability in the diagnosis of pleural effusions than chest X-ray can be repeated serially at the bedside without any radiation risk. One hundred and fifty patients after cardiac surgery with basal pleural opacity on chest X-ray have been included in our prospective observational study during a two-year period. Effusion was confirmed on postoperative day (POD) 5.9+/-3.2 per chest ultrasound sonography. Inclusion criteria for subsequent thoracentesis based on clinical grounds alone and were not protocol-driven. Major inclusion criteria were: dyspnea and peripheral oxygen saturation (SpO(2)) levels < or = 92% and the maximal distance between mid-height of the diaphragm and visceral pleura (D > or = 30 mm). One hundred and thirty-five patients (90%) were drained with a 14-G needle if according to the simplified formula: V (ml)=[16 x D (mm)] the volume of the pleural effusion was around 500 ml. The success rate of obtaining fluid was 100% without any complications. There is a high accuracy between the estimated and drained pleural effusion. Simple quantification of pleural effusion enables time and cost-effective decision-making for thoracentesis in postoperative patients. PMID- 19903688 TI - Aortocoronary bypass graft fistula after surgical treatment of circumflex coronary artery fistula: a unique variation of a rare condition successfully treated with percutaneous embolization. AB - Multiple coronary artery fistulae are rare, complications can be life threatening, and with large or symptomatic fistulae, intervention is mandatory. Both surgical and percutaneous interventions are well-described. We believe this is the first report of the embolization of an acquired fistula following initial surgical treatment of multiple congenital fistulae. PMID- 19903689 TI - Proposed disease activity criteria for mixed connective tissue disease. PMID- 19903690 TI - Regulation of N-type voltage-gated calcium channels (Cav2.2) and transmitter release by collapsin response mediator protein-2 (CRMP-2) in sensory neurons. AB - Collapsin response mediator proteins (CRMPs) mediate signal transduction of neurite outgrowth and axonal guidance during neuronal development. Voltage-gated Ca(2+) channels and interacting proteins are essential in neuronal signaling and synaptic transmission during this period. We recently identified the presynaptic N-type voltage-gated Ca(2+) channel (Cav2.2) as a CRMP-2-interacting partner. Here, we investigated the effects of a functional association of CRMP-2 with Cav2.2 in sensory neurons. Cav2.2 colocalized with CRMP-2 at immature synapses and growth cones, in mature synapses and in cell bodies of dorsal root ganglion (DRG) neurons. Co-immunoprecipitation experiments showed that CRMP-2 associates with Cav2.2 from DRG lysates. Overexpression of CRMP-2 fused to enhanced green fluorescent protein (EGFP) in DRG neurons, via nucleofection, resulted in a significant increase in Cav2.2 current density compared with cells expressing EGFP. CRMP-2 manipulation changed the surface levels of Cav2.2. Because CRMP-2 is localized to synaptophysin-positive puncta in dense DRG cultures, we tested whether this CRMP-2-mediated alteration of Ca(2+) currents culminated in changes in synaptic transmission. Following a brief high-K(+)-induced stimulation, these puncta became loaded with FM4-64 dye. In EGFP and neurons expressing CRMP-2-EGFP, similar densities of FM-loaded puncta were observed. Finally, CRMP-2 overexpression in DRG increased release of the immunoreactive neurotransmitter calcitonin gene-related peptide (iCGRP) by approximately 70%, whereas siRNA targeting CRMP-2 significantly reduced release of iCGRP by approximately 54% compared with control cultures. These findings support a novel role for CRMP-2 in the regulation of N-type Ca(2+) channels and in transmitter release. PMID- 19903691 TI - Eeyarestatin I inhibits Sec61-mediated protein translocation at the endoplasmic reticulum. AB - Production and trafficking of proteins entering the secretory pathway of eukaryotic cells is coordinated at the endoplasmic reticulum (ER) in a process that begins with protein translocation via the membrane-embedded ER translocon. The same complex is also responsible for the co-translational integration of membrane proteins and orchestrates polypeptide modifications that are often essential for protein function. We now show that the previously identified inhibitor of ER-associated degradation (ERAD) eeyarestatin 1 (ES(I)) is a potent inhibitor of protein translocation. We have characterised this inhibition of ER translocation both in vivo and in vitro, and provide evidence that ES(I) targets a component of the Sec61 complex that forms the membrane pore of the ER translocon. Further analyses show that ES(I) acts by preventing the transfer of the nascent polypeptide from the co-translational targeting machinery to the Sec61 complex. These results identify a novel effect of ES(I), and suggest that the drug can modulate canonical protein transport from the cytosol into the mammalian ER both in vitro and in vivo. PMID- 19903693 TI - Arabidopsis Rab-E GTPases exhibit a novel interaction with a plasma-membrane phosphatidylinositol-4-phosphate 5-kinase. AB - Rab GTPases of the Arabidopsis Rab-E subclass are related to mammalian Rab8 and are implicated in membrane trafficking from the Golgi to the plasma membrane. Using a yeast two-hybrid assay, Arabidopsis phosphatidylinositol-4-phosphate 5 kinase 2 (PtdIns(4)P 5-kinase 2; also known as PIP5K2), was shown to interact with all five members of the Rab-E subclass but not with other Rab subclasses residing at the Golgi or trans-Golgi network. Interactions in yeast and in vitro were strongest with RAB-E1d[Q74L] and weakest with the RAB-E1d[S29N] suggesting that PIP5K2 interacts with the GTP-bound form. PIP5K2 exhibited kinase activity towards phosphatidylinositol phosphates with a free 5-hydroxyl group, consistent with PtdIns(4)P 5-kinase activity and this activity was stimulated by Rab binding. Rab-E proteins interacted with PIP5K2 via its membrane occupancy and recognition nexus (MORN) domain which is missing from animal and fungal PtdIns(4)P 5-kinases. In plant cells, GFP:PIP5K2 accumulated at the plasma membrane and caused YFP:RAB-E1d to relocate there from its usual position at the Golgi. GFP:PIP5K2 was rapidly turned over by proteasomal activity in planta, and overexpression of YFP:PIP5K2 caused pleiotropic growth abnormalities in transgenic Arabidopsis. We propose that plant cells exhibit a novel interaction in which PIP5K2 binds GTP-bound Rab-E proteins, which may stimulate temporally or spatially localized PtdIns(4,5)P(2) production at the plasma membrane. PMID- 19903694 TI - Differences in etiological and clinical manifestations in upper extremity and lower limb deep venous thrombosis patients from India. AB - We assessed the clinical manifestations in upper and lower limb deep venous thrombosis patients from India and difference in etiological factors. Fifty-three patients with primary upper extremity deep vein thrombosis (UEDVT; males 30, females 23) and 236 patients with lower limb deep vein thrombosis (LLDVT; males 157, females 79) were included in this study. The thrombophilia markers studied were protein C (PC), protein S (PS), antithrombin (AT) III, and factor V Leiden (FVL) mutation. Females had significantly higher prevalence of prothrombotic markers as compared to males (P = .046) in the UEDVT group. No statistically significant differences in the prevalence of prothrombotic markers were observed between the LLDVT and the UEDVT patients. The clinical picture however revealed greater involvement of thrombus of the iliofemoral vein (P = .009) and the proximal tibial vein (P = .005) in males than females, while no differences were observed in the clinical manifestations between the 2 sexes in UEDVT patients. Our study is able to give a broad perspective of the prevalence data of UEDVT and LLDVT in the city of Mumbai of approximately 5 million population served by this hospital. We conclude that the topology of thrombosis in deep vein thrombosis (DVT) patients in India is different from that of the Western countries. PMID- 19903692 TI - The integrin adhesion complex changes its composition and function during morphogenesis of an epithelium. AB - Cell adhesion to the extracellular matrix (ECM) is mediated by the integrin family of transmembrane receptors. Integrins link ECM ligands to the cytoskeleton, providing strong attachment to enable cell-shape change and tissue integrity. This connection is made possible by an intracellular complex of proteins, which links to actin filaments and controls signalling cascades that regulate cytoskeletal rearrangements. We have identified stress-fibre-associated focal adhesions that change their composition during tissue morphogenesis. Early expression of alphaPS1betaPS integrin decreases the levels of the actin nucleating factors Enabled, Diaphanous and profilin, as well as downregulating the amount of F-actin incorporated into the stress fibres. As follicle cells mature in their developmental pathway and become squamous, the integrin in the focal adhesions changes from alphaPS1betaPS to alphaPS2betaPS. During the switch, stress fibres increase their length and change orientation, first changing by 90 degrees and then reorienting back. The normal rapid reorientation requires new expression of alphaPS2betaPS, which also permits recruitment of the adaptor protein tensin. Unexpectedly, it is the extracellular portion of the alphaPS2 subunit that provides the specificity for intracellular recruitment of tensin. Molecular variation of the integrin complex is thus a key component of developmentally programmed morphogenesis. PMID- 19903695 TI - Phosphatidylethanolamine at the luminal endothelial surface--implications for hemostasis and thrombotic autoimmunity. AB - OBJECTIVE: Accumulating evidence suggests that phosphatidylethanolamine (PE) is physically present at the luminal endothelial surface, where it tentatively functions as a critical anticoagulant. The goal of the current investigation was 3-fold: to characterize the distribution profile of PE at the luminal endothelial surface; to examine the immunoreactivity to the vascular endothelium by anti-PE (aPE) sera from patients presenting with thrombosis; and to discuss the potential mechanism of PE upregulation by endothelial cells. METHODS: The rat aortic arch was selected as major conduit vessel under significant hemodynamic burden. The presence of PE and the antigenic profile of aPE sera at the luminal endothelial surface were examined using duramycin as a PEbinding probe and immunohistochemistry. Phosphatidylethanolamine upregulation at endothelial cell surface was investigated using cultured monolayer subject to laminar shear stress or thrombin treatment. RESULTS: High levels of PE were detected at the luminal endothelial surface of aortic flow dividers, the ascending aorta, and the outer curvature of the aortic arch. The antigenic profiles of aPE sera, which are highly associated with elevated thrombotic risks in patients, are consistent with PE distribution along the endothelial surface. Finally, PE is upregulated at the surface of cultured endothelial cells in response to luminal shear stress but not thrombin. CONCLUSIONS: The current data describe the physical distribution of vascular PE at the blood-endothelium interface. The luminal PE presents a vulnerability to anti-PE autoimmunity and is consistent with the association between aPE and elevated risk for idiopathic thrombosis. PMID- 19903696 TI - Incidence and clinical importance of lupus anticoagulant in children with recurrent upper respiratory tract infection. AB - BACKGROUND: This study aims to understand the incidence and presence of lupus anticoagulant (LA) in children with recurrent upper respiratory tract infection (URTI). METHODS: One hundred and sixty-five patients who were admitted to Departments of Pediatrics and Otolaryngology at Ege University Faculty of Medicine during the last 2 years and 120 age-matched healthy children as a control group were enrolled in the study. RESULTS: The presence of LA in serum was positive in 8 (4.8%) cases in the patient group while only 2 (1.6%) cases in the healthy control group (P = .03). Mean age of patients with LA positive was significantly lower than those of negative cases (P = .02). Of the patients, 92 (55.8%) had adenoid hypertrophy. The annual frequency of URTI did not differ significantly between the LA patients and the LA-negative patients (7.5/year and 6.9/year, respectively). None of the patients with LA positive had adenoid hypertrophy (P = .009). Activated partial thromboplastin time was prolonged in 6 (3.6%) of 165 patients. Of these 6 patients, 2 were also LA positive. The presence of LA disappeared in all the 8 patients 2 months after the diagnosis. Lupus anticoagulant was found negative in all patients at the end of the second month. CONCLUSION: We found that the ratio of the presence of LA is higher in children with recurrent URTI than healthy children. However, the presence of LA does not lead to bleeding and/or thrombosis, and it disappears in a short period of time. PMID- 19903697 TI - Human platelet aggregation and degranulation is induced in vitro by L-thyroxine, but not by 3,5,3'-triiodo-L-thyronine or diiodothyropropionic acid (DITPA). AB - The endogenous thyroid hormones L-thyroxine (T(4)) and 3,5,3'-triiodo-L-thyronine (T(3)) induce angiogenesis via an endothelial cell iodothyronine receptor on integrin alphaVbeta3. This receptor also exists on platelets. Diiodothyropropionic acid (DITPA) and GC-1, a noniodinated thyroid hormone analog, also induce angiogenesis. Here we examined the effects of iodothyronines (L-T(4) vs L-T(3)) and analogs DITPA and GC-1 on human platelet function. Subthreshold aggregation of platelets obtained from healthy human donors was induced with collagen. Platelet activation (proaggregation) and adenosine triphosphate (ATP) secretion (degranulation) induced by L-T( 4), L-T(4)-agarose, L-T(3), DITPA, or GC-1 were determined simultaneously. Platelet aggregation and ATP secretion induced by a subthreshold level of collagen were enhanced 3-fold by either L-T(4) or L-T( 4)-agarose (0.01 micromol/L) as compared to control, whereas, L-T( 3), DITPA, or GC-1 had no effect under the same conditions. The platelet proaggregatory and degranulation effects of L-T(4) were blocked by the alphavbeta3 antagonist XT199 (0.1 micromol/ L) and by tetraiodothyroacetic acid (tetrac; 0.1 micromol/L). Tetrac inhibits binding of thyroid hormone analogs to the receptor on alphavbeta3 and lacks thyromimetic activity at this site; thus, the proaggregatory action of L-T(4) likely involves the cell surface receptor on integrin alphavbeta3. The thyroid hormone receptor (TR) on human platelets but not endothelial cells distinguishes among iodothyronines, reflecting quantitative differences in integrin sites on endothelial cells and platelets or qualitative differences in the phospholipids/protein microenvironment of endothelial and platelet membranes that can affect integrin function. Additional studies in different populations with larger sample sizes are warranted to determine the impact of the current findings on clinical interventions. PMID- 19903698 TI - Steroid-induced iatrogenic disease after treating for pseudothrombocytopenia. AB - Pseudothrombocytopenia, a spontaneous in vitro occurrence after the addition of anticoagulant to blood, causes clumping of platelets resulting in a spurious observation of low platelet counts (<10,000/MUL) without any associated hemorrhagic manifestations. We describe a 46-year-old male patient who was diagnosed with immune thrombocytopenic purpura (ITP) based on a reported platelet count of 22,000/MUL. He was prescribed high-dose glucocorticoid therapy, up to 60 mg of prednisolone daily for over a year. After repeated hospital admissions, he came under our care as an emergency admission for nonketotic hyperosmolar hyperglycemia. He was diabetic, osteopenic, and had been treated for tuberculosis, all likely consequences of prolonged glucocorticoid therapy. In the presence of persistent platelet counts below 10,000/MUL, and without associated clinical hematological manifestations of ITP, a smear of citrated blood was examined and a platelet count of 215,000/MUL was observed. This case highlights the possible consequences of misdiagnosis of pseudothrombocytopenia. Failure to recognize this phenomenon may lead to debilitating iatrogenic disease. PMID- 19903699 TI - Oxidative modification of fibrinogen affects its binding activity to glycoprotein (GP) IIb/IIIa. AB - AIM: Proteins are sensitive biomarkers of human disease condition associated with oxidative stress. Alteration of protein structures by oxidants may result in partial or complete loss of protein functions. We have investigated the effect of structural modifications induced by metal ion catalyzed oxidation of fibrinogen on its binding capacity to glycoprotein IIb/IIIa (GpIIb/IIIa) and human platelets. METHODS: We identified and quantified of binding capacity of native and oxidized fibrinogen to its receptor in vitro by flow cytometer. Dityrosine formation on oxidized fibrinogen were detected spectrophotometrically. Elevated degradation products of fibrinogen after oxidation were revealed in the HPLC analysis. The native and oxidized fibrinogen were analyzed on mass spectrum upon digestion with trypsin. RESULTS: Oxidatively modified fibrinogen showed less binding activity than native fibrinogen to GpIIb/IIIa coated micro beads and human platelets whereas slightly higher binding capacity to ADP induced stimulated platelets. Formation of di-tyrosines in the amino acid side chains of fibrinogen were observed upon oxidation. Decreased binding capacity of oxidized fibrinogen correlated with intensities of dityrosine formation. Oxidized fibrinogen had more ion-mass intensities at higher than native fibrinogen. CLINICAL IMPLICATIONS: Important point is decreased of binding capacity of the oxidized fibrinogen to own receptor. The decreased rate of binding, leading to effect in the diseases of clot formation may account for the association between oxidation of fibrinogen and the incidence of effect in human diseases. PMID- 19903700 TI - PPARdelta and PGC1alpha act cooperatively to induce haem oxygenase-1 and enhance vascular endothelial cell resistance to stress. AB - AIMS: Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear receptor superfamily of ligand-activated transcriptional regulators. PPARdelta has an established role in metabolism, wound healing, and angiogenesis. However, little is known about its function in endothelial homeostasis. We investigated the role of PPARdelta and its co-activator, PPARgamma co-activator 1alpha (PGC1alpha), in vasculoprotection against oxidant-induced injury via induction of haem oxygenase-1. METHODS AND RESULTS: En face confocal microscopy of murine aortas demonstrated that the PPARdelta-selective ligand GW501516 induced endothelial haem oxygenase-1 expression. In vitro PPARdelta ligands induced a significant increase in haem oxygenase-1 mRNA, protein, and enzyme activity, resulting in enhanced human endothelial cell protection against cellular stress induced by hydrogen peroxide or leptin. Moreover, adenoviral mediated overexpression of haem oxygenase-1 increased PPARdelta promoter activity and mRNA levels, amplifying the effect of PPARdelta ligands through a positive feedback loop. Mutation of PPAR response element binding sites in the haem oxygenase-1 promoter/enhancer region revealed haem oxygenase-1 to be a direct PPARdelta target gene. Inhibition of either haem oxygenase-1 or PPARdelta abrogated PPARdelta ligand-induced endothelial cytoprotection. Furthermore, siRNA depletion of PGC1alpha demonstrated that this co-regulator acts as an essential PPARdelta transcriptional co-activator for haem oxygenase-1 induction by PPARdelta ligands and its subsequent cytoprotective actions. CONCLUSION: We have identified an important relationship between PPARdelta, PGC1alpha, and haem oxygenase-1, demonstrating that haem oxygenase-1 induction plays an important role in cytoprotective actions of PPARdelta ligands in vascular endothelium. In light of the protective effects of haem oxygenase-1 against atherogenesis, we suggest that PPARdelta represents a potentially important therapeutic target in the vasculature. PMID- 19903701 TI - Progestin regulates chemokine (C-X-C motif) ligand 14 transcript level in human endometrium. AB - Leukocyte populations change profoundly in the human endometrium during the menstrual cycle. However the predominant cell, the uterine natural killer (uNK) cell does not contain steroid receptors. From gene array analysis we identified a transcript encoding chemokine (C-X-C motif) ligand 14 (CXCL14) which is markedly up-regulated in the secretory phase of the cycle. We confirm this data by northern blotting and quantitative PCR. Using in situ hybridization we localized CXCL14 mRNA to the glandular epithelial cells where it was detected only in the secretory phase of the cycle. Candidate progesterone response elements were identified at positions -2028/-2007 and -722/-697 (PRE1 and PRE2, respectively) relative to the translation start site. These were functionally tested using luciferase reporter deletion constructs, electrophoretic mobility shift assays and site-directed mutagenesis. The deletion/mutation of these sites reduced progesterone induction by 40 and 20%, respectively. Finally, we demonstrated that recombinant CXCL14 stimulated uNK cell chemotaxis in vitro. We therefore conclude that CXCL14 is likely to be regulated by progesterone in human endometrium and that it may exert a chemoattractive effect on uNK cells and in part be responsible for their clustering around the epithelial glands. PMID- 19903702 TI - Is haemoglobin A1c a step forward for diagnosing diabetes? PMID- 19903703 TI - Cervical radiculopathy. Never mind the treatment, what about the tests? PMID- 19903704 TI - Propofol is safely and widely used in emergency departments. PMID- 19903706 TI - Risks at new academic year. Solutions to the August problem. PMID- 19903707 TI - Risks at new academic year. Study may not apply to the UK. PMID- 19903708 TI - Great Danes. It's a Nordic thing. PMID- 19903709 TI - The WMA speaks out on Iran but not on Israel. Why not? PMID- 19903711 TI - Flu vaccine and egg allergy. About available flu vaccines. PMID- 19903712 TI - Future of flu vaccines. Please may we have an RCT now? PMID- 19903713 TI - Future of flu vaccines. Expediting clinical trials in a pandemic. PMID- 19903716 TI - Battle against hospital acquired infections has been too limited, MPs' report says. PMID- 19903717 TI - Induction of spermatogenic cell apoptosis in prepubertal rat testes irrespective of testicular steroidogenesis: a possible estrogenic effect of di(n-butyl) phthalate. AB - Although di(n-butyl) phthalate (DBP), a suspected endocrine disruptor, induces testicular atrophy in prepubertal male rats, whether it exerts estrogenic activity in vivo remains a matter of debate. In the present study, we explored the estrogenic potency of DBP using 3-week-old male rats, and then examined the relationship between estrogen-induced spermatogenic cell apoptosis and testicular steroidogenesis. Daily exposure to DBP for 7 days caused testicular atrophy due to loss of spermatogenic cells, whereas testicular steroidogenesis was almost the same with the control values. A single exposure of DBP decreased testicular steroidogenesis in addition to decreasing the level of serum LH at 3 h after DBP treatment, with an extremely high incidence of apoptotic spermatogenic cells at 6 h after administration. To elucidate the estrogenic activity of DBP, we carried out an inhibition study using pure antiestrogen ICI 182,780 (ICI) in a model of spermatogenic cell apoptosis induced by DBP or estradial-3-benzoate (EB). Although both the DBP- and EB-treated groups showed a significant increase in spermatogenic cell apoptosis, ICI pretreatment significantly decreased the number of apoptotic spermatogenic cells in these two groups. In contrast, testicular steroidogenesis and serum FSH were significantly reduced in all the treated groups, even in the DBP+ICI and EB+ICI groups. Taken together, these findings led us to conclude that estrogenic compounds such as DBP and EB induce spermatogenic cell apoptosis in prepubertal rats, probably by activating estrogen receptors in testis, and that reduction in testicular steroidogenic function induced by estrogenic compounds is not associated with spermatogenic cell apoptosis. PMID- 19903718 TI - About tyrosine kinase inhibitors (TKIs) in prostate cancer: where do we go from here? PMID- 19903719 TI - Does enfuvirtide increase the risk of bacterial pneumonia in patients receiving combination antiretroviral therapy? AB - BACKGROUND: Pooled analysis of the TORO comparative clinical trial data sets showed a significantly higher incidence rate (IR) of bacterial pneumonia (BP) among patients treated with enfuvirtide-containing combination antiretroviral therapy (ENF-cART) than in those treated with other cART regimens. OBJECTIVES: To examine the possible impact of ENF-cART on the risk of BP. METHODS: From the French Hospital Database on HIV, we selected two groups of patients among cART treated patients who were prescribed a new cART regimen during the period 2001 2006, when their CD4 counts were <350 cells/mm(3). The ENF-cART and cART groups consisted of 1220 and 9374 patients, respectively. Poisson regression models were used to quantify the relationship between ENF-cART therapy and the risk of BP. RESULTS: At baseline the median CD4 counts were 100 and 211 cells/mm(3) and the median plasma viral load (pVL) values were 60 276 and 2702 copies/mL in the ENF cART and cART groups, respectively. The respective BP IRs were 0.65 [95% confidence interval (CI) 0.25-1.06] and 0.31 (95% CI 0.25-0.38) cases per 100 person-years. After adjustment for age, the HIV transmission group, the time period, co-trimoxazole prophylaxis, and stratified CD4 cell counts and pVL values, we found that the BP risk ratio was not increased by enfuvirtide treatment (relative rate 1.39; 95% CI 0.46-4.13). In contrast, lower CD4 cell counts and higher pVL values were significantly associated with a higher risk of BP. CONCLUSIONS: ENF-cART is not associated with a significantly higher risk of BP than other cART regimens, although the value of the adjusted risk and the upper limit of the CI do not allow us to exclude a small increased risk. PMID- 19903720 TI - Viral resistance to specifically targeted antiviral therapies for hepatitis C (STAT-Cs). AB - Promising results have been observed with an investigational drug class for hepatitis C (HCV), the specifically targeted antiviral therapies for hepatitis C (STAT-Cs), when combined with peginterferon plus ribavirin (Peg-IFN/RBV). This class has the potential to increase sustained virological response (SVR) rates and reduce therapy duration in genotype 1 chronic HCV patients compared with Peg IFN/RBV alone. However, because of the remarkable sequence variation in HCV (resulting from the high viral replication rate and intrinsically error-prone nature of HCV polymerase), variants with reduced susceptibility to STAT-Cs can occur naturally before treatment, usually at low levels, and can be selected in patients not responding to potent STAT-C treatment. This review first describes how resistance to a STAT-C can develop and then provides an overview of mutations that confer varying levels of resistance to STAT-Cs, which have been identified and characterized using both genotypic and phenotypic tools. We will discuss why an understanding of the selection of variants with reduced susceptibility to a treatment regimen may be important in optimizing the use of this new class of HCV therapy. Strategies for optimizing treatment regimens to increase response rates, and thereby minimize resistance, will be discussed. Finally, although resistance can be a consequence of not achieving an SVR on an initial regimen, there may be alternative treatment options for patients to achieve an SVR in the future. Future potential therapeutic strategies to address patients who do develop resistance to STAT-Cs are discussed, including combination therapy with multiple STAT-Cs with non-overlapping resistance profiles. PMID- 19903721 TI - The impact of pediatric chronic pain on parents' health-related quality of life and family functioning: reliability and validity of the PedsQL 4.0 Family Impact Module. AB - OBJECTIVE: To evaluate the psychometric properties of the Family Impact Module (FIM), a parent self-report measure of health-related quality of life (HRQOL) and family functioning, among parents of youth with chronic pain. METHODS: Parents (N = 458) completed the FIM (Total Impact, HRQOL, and Family Functioning scales); parents and youth (N = 332) completed measures of pain catastrophizing, pediatric quality of life, and emotional/behavioral functioning. RESULTS: The FIM demonstrated strong internal consistency and item-total correlations. All FIM scales were positively associated with pain catastrophizing, functional disability, and emotional/behavioral problems; and inversely related to pediatric quality of life. Mothers reported significantly worse HRQOL than fathers. Mothers and fathers did not differ on reports of Family Functioning. HRQOL and Family Functioning did not differ as a function of pain diagnosis. CONCLUSION: The FIM appears to be a suitable measure of parent self-reported HRQOL and family functioning in pediatric chronic pain. PMID- 19903723 TI - Adherence to the Mediterranean diet and risk of coronary heart disease in the Spanish EPIC Cohort Study. AB - No known cohort study has investigated whether the Mediterranean diet can reduce incident coronary heart disease (CHD) events in a Mediterranean population. This study examined the relation between Mediterranean diet adherence and risk of incident CHD events in the 5 Spanish centers of the European Prospective Investigation into Cancer and Nutrition. Analysis included 41,078 participants aged 29-69 years, recruited in 1992-1996 and followed up until December 2004 (mean follow-up:10.4 years). Confirmed incident fatal and nonfatal CHD events were analyzed according to Mediterranean diet adherence, measured by using an 18 unit relative Mediterranean diet score. A total of 609 participants (79% male) had a fatal or nonfatal confirmed acute myocardial infarction (n = 468) or unstable angina requiring revascularization (n = 141). After stratification by center and age and adjustment for recognized CHD risk factors, high compared with low relative Mediterranean diet score was associated with a significant reduction in CHD risk (hazard ratio = 0.60, 95% confidence interval: 0.47, 0.77). A 1-unit increase in relative Mediterranean diet score was associated with a 6% reduced risk of CHD (95% confidence interval: 0.91, 0.97), with similar risk reductions by sex. Mediterranean diet adherence was associated with a significantly reduced CHD risk in this Mediterranean country, supporting its role in primary prevention of CHD in healthy populations. PMID- 19903724 TI - Importance of routine public health influenza surveillance: detection of an unusual W-shaped influenza morbidity curve. AB - Seasonal influenza causes excess morbidity and mortality at the extremes of age: It disproportionately affects the very young and the very old, typically resulting in "U"-shaped age-distributed curves. By means of a well-established public health department surveillance system using positive influenza tests submitted from sentinel sites, the authors generated annual influenza-specific morbidity curves over a 10-year period (1998-2008) for St. Louis County, Missouri. The authors detected an unusually high incidence of cases of medically attended test-positive influenza, particularly in young adults, during the 2007 2008 season, resulting in an unexpected "W"-shaped age-distributed morbidity curve that was distinctly unique in comparison with the prior 9 influenza seasons. Public health influenza surveillance programs are useful tools for detecting emerging epidemiologic trends that may have clinical importance. PMID- 19903725 TI - Current concepts in the assessment and treatment of hepatic encephalopathy. AB - Hepatic encephalopathy (HE) is defined as a metabolically induced, potentially reversible, functional disturbance of the brain that may occur in acute or chronic liver disease. Standardized nomenclature has been proposed but a standardized approach to the treatment, particularly of persistent, episodic and recurrent encephalopathy associated with liver cirrhosis has not been proposed. This review focuses on the pathogenesis and treatment of HE in patients with cirrhosis. The pathogenesis and treatment of hepatic encephalopathy in fulminant hepatic failure is quite different and is reviewed elsewhere. PMID- 19903726 TI - The pea, the yeast and the prostate. PMID- 19903727 TI - A slowly expanding intracranial lesion. PMID- 19903728 TI - Comment on: Web resources for rare auto-inflammatory diseases: towards a common patient registry. PMID- 19903729 TI - Factors affecting the shear bond strength of metal and ceramic brackets bonded to different ceramic surfaces. AB - The aims of this study were to evaluate the shear bond strength (SBS) of metal and ceramic brackets bonded to two different all-ceramic crowns, IPS Empress 2 and In-Ceram Alumina, to compare the SBS between hydrofluoric acid (HFA), phosphoric acid etched, and sandblasted, non-etched all-ceramic surfaces. Ninety six all-ceramic crowns were fabricated resembling a maxillary left first premolar. The crowns were divided into eight groups: (1) metal brackets bonded to sandblasted 9.6 per cent HFA-etched IPS Empress 2 crowns; (2) metal brackets bonded to sandblasted 9.6 per cent HFA-etched In-Ceram crowns; (3) ceramic brackets bonded to sandblasted 9.6 per cent HFA-etched IPS Empress 2 crowns; (4) ceramic brackets bonded to sandblasted 9.6 per cent HFA-etched In-Ceram crowns; (5) metal brackets bonded to sandblasted 37 per cent phosphoric acid-etched IPS Empress 2 crowns; (6) metal brackets bonded to sandblasted 37 per cent phosphoric acid-etched In-Ceram crowns; (7) metal brackets bonded to sandblasted, non-etched IPS Empress 2 crowns; and (8) metal brackets bonded to sandblasted, non-etched In Ceram crowns. Metal and ceramic orthodontic brackets were bonded using a conventional light polymerizing adhesive resin. An Instron universal testing machine was used to determine the SBS at a crosshead speed of 0.1 mm/minute. Comparison between groups was performed using a univariate general linear model and chi-squared tests. The highest mean SBS was found in group 3 (120.15 +/- 45.05 N) and the lowest in group 8 (57.86 +/- 26.20 N). Of all the variables studied, surface treatment was the only factor that significantly affected SBS (P < 0.001). Acid etch application to sandblasted surfaces significantly increased the SBS in groups 1, 2, 5, and 6. The SBS of metal brackets debonded from groups 1, 3, and 5 were not significantly different from those of groups 2, 4, and 6. All debonded metal brackets revealed a similar pattern of bond failure at the adhesive-restorative interface. However, ceramic brackets had a significantly different adhesive failure pattern with dominant failure at the adhesive-bracket interface. Ceramic fractures after bracket removal were found more often in groups 1-4. No significant difference in ceramic fracture was observed between the IPS Empress 2 and In-Ceram groups. PMID- 19903730 TI - Silencing Sl-EBF1 and Sl-EBF2 expression causes constitutive ethylene response phenotype, accelerated plant senescence, and fruit ripening in tomato. AB - The hormone ethylene regulates a wide range of plant developmental processes and EBF (EIN3-binding F-box) proteins were shown to negatively regulate the ethylene signalling pathway via mediating the degradation of EIN3/EIL proteins. The present study reports on the identification of two tomato F-box genes, Sl-EBF1 and Sl-EBF2 from the EBF subfamily. The two genes display contrasting expression patterns in reproductive and vegetative tissues and in response to ethylene and auxin treatment. Sl-EBF1 and Sl-EBF2 genes are actively regulated at crucial stages in the development of the reproductive organs. Their dynamic expression in flowers during bud-to-anthesis and anthesis-to-post-anthesis transitions, and at the onset of fruit ripening, suggests their role in situations where ethylene is required for stimulating flower opening and triggering fruit ripening. VIGS mediated silencing of a single tomato EBF gene uncovered a compensation mechanism that tends to maintain a threshold level of Sl-EBF expression via enhancing the expression of the second Sl-EBF gene. In line with this compensation, tomato plants silenced for either of the Sl-EBF genes were indistinguishable from control plants, indicating functional redundancy among Sl-EBF genes. By contrast, co-silencing of both Sl-EBFs resulted in ethylene-associated phenotypes. While reports on EBF genes to date have focused on their role in modulating ethylene responses in Arabidopsis, the present study uncovered their role in regulating crucial stages of flower and fruit development in tomato. The data support the hypothesis that protein degradation via the ubiquitin/26S proteasome pathway is a control point of fruit ripening and open new leads for engineering fruit quality. PMID- 19903731 TI - The enigma of Gerstmann's syndrome revisited: a telling tale of the vicissitudes of neuropsychology. AB - Eighty years ago, the Austrian neurologist Josef Gerstmann observed in a few patients a concomitant impairment in discriminating their own fingers, writing by hand, distinguishing left from right and performing calculations. He claimed that this tetrad of symptoms constituted a syndromal entity, assigned it to a lesion of the dominant parietal lobe and suggested that it was due to damage of a common functional denominator. Ever since, these claims have been debated and an astute synopsis and sceptical discussion was presented 40 years ago by MacDonald Critchley in this journal. Nonetheless, Gerstmann's syndrome has continued to intrigue both clinical neurologists and researchers in neuropsychology, and more frequently than not is described in textbooks as an example of parietal lobe damage. In this review, we revisit the chequered history of this syndrome, which can be seen as a case study of the dialectic evolution of concepts in neuropsychology. In light of several modern era findings of pure cases we conclude that it is legitimate to label the conjunction of symptoms first described by Gerstmann as a 'syndrome', but that it is very unlikely that damage to the same population of cortical neurons should account for all of the four symptoms. Instead, we propose that a pure form of Gerstmann's syndrome might arise from disconnection, via a lesion, to separate but co-localized fibre tracts in the subcortical parietal white matter, a hypothesis for which we have recently provided evidence using combined imaging of functional and structural organization in the healthy brain. PMID- 19903732 TI - Executive function and fluid intelligence after frontal lobe lesions. AB - Many tests of specific 'executive functions' show deficits after frontal lobe lesions. These deficits appear on a background of reduced fluid intelligence, best measured with tests of novel problem solving. For a range of specific executive tests, we ask how far frontal deficits can be explained by a general fluid intelligence loss. For some widely used tests, e.g. Wisconsin Card Sorting, we find that fluid intelligence entirely explains frontal deficits. When patients and controls are matched on fluid intelligence, no further frontal deficit remains. For these tasks too, deficits are unrelated to lesion location within the frontal lobe. A second group of tasks, including tests of both cognitive (e.g. Hotel, Proverbs) and social (Faux Pas) function, shows a different pattern. Deficits are not fully explained by fluid intelligence and the data suggest association with lesions in the right anterior frontal cortex. Understanding of frontal lobe deficits may be clarified by separating reduced fluid intelligence, important in most or all tasks, from other more specific impairments and their associated regions of damage. PMID- 19903734 TI - Brain alpha-synuclein accumulation in multiple system atrophy, Parkinson's disease and progressive supranuclear palsy: a comparative investigation. AB - Alpha-synuclein is a major component of Lewy bodies and glial cytoplasmic inclusions, pathological hallmarks of idiopathic Parkinson's disease and multiple system atrophy, and it is assumed to be aetiologically involved in these conditions. However, the quantitative status of brain alpha-synuclein in different Parkinsonian disorders is still unresolved and it is uncertain whether alpha-synuclein accumulation is restricted to regions of pathology. We compared membrane-associated, sodium dodecyl sulfate-soluble alpha-synuclein, both the full-length 17 kDa and high molecular weight species, by western blotting in autopsied brain of patients with Parkinson's disease (brainstem-predominant Lewy body disease: n = 9), multiple system atrophy (n = 11), progressive supranuclear palsy (n = 16), and of normal controls (n = 13). Brain of a patient with familial Parkinsonism-dementia due to alpha-synuclein locus triplication (as positive control) showed increased membrane-associated, sodium dodecyl sulfate-soluble alpha-synuclein levels with abundant high molecular weight immunoreactivity. In multiple system atrophy, a massive increase in 17 kDa membrane-associated, sodium dodecyl sulfate-soluble alpha-synuclein was observed in highly pathologically affected regions, including putamen (+1760%, range +625-2900%), substantia nigra [+1000% (+356-1850%)], and white matter of internal capsule [+2210% (+430-6830%)] together with numerous high molecular weight species. Levels of 17 kDa membrane associated, sodium dodecyl sulfate-soluble alpha-synuclein were only modestly increased in less affected areas (cerebellar cortex, +95%; caudate, +30%; with both also showing numerous high molecular weight species) and were generally normal in cerebral cortices. In both Parkinson's disease and progressive supranuclear palsy, membrane-associated, sodium dodecyl sulfate-soluble alpha synuclein levels were normal in putamen and frontal cortex whereas a trend was observed for variably increased 17 kDa membrane-associated, sodium dodecyl sulfate-soluble alpha-synuclein concentrations [+184% (-60% to +618%)] with additional high molecular weight species in Parkinson's disease substantia nigra. No obvious correlation was observed between nigral membrane-associated, sodium dodecyl sulfate-soluble alpha-synuclein accumulation and Lewy body density in Parkinson's disease. Two progressive supranuclear palsy cases had membrane associated, sodium dodecyl sulfate-soluble alpha-synuclein accumulation in substantia nigra similar to multiple system atrophy. Several Parkinson's disease patients had very modest high molecular weight membrane-associated, sodium dodecyl sulfate-soluble alpha-synuclein accumulation in putamen. Levels of 17-kDa membrane-associated, sodium dodecyl sulfate-soluble alpha-synuclein were generally positively correlated with those of high molecular weight membrane associated, sodium dodecyl sulfate-soluble alpha-synuclein and there was a trend for a positive correlation between striatal dopamine loss and 17-kDa membrane associated, sodium dodecyl sulfate-soluble alpha-synuclein concentrations in multiple system atrophy. Brain membrane-associated, sodium dodecyl sulfate soluble alpha-synuclein accumulations in Parkinson's disease and multiple system atrophy are regionally specific, suggesting that these sporadic alpha synucleinopathies, unlike familial Parkinsonism-dementia, are not associated with a simple global over-expression of the protein. Despite a similar extent of dopamine depletion, the magnitude of brain membrane-associated, sodium dodecyl sulfate-soluble alpha-synuclein changes is disease specific, with multiple system atrophy clearly having the most severe accumulation. Literature discrepancies on alpha-synuclein status in 'Parkinson's disease' might be explained by inclusion of cases not having classic brainstem-predominant Lewy body disease and by variable alpha-synuclein accumulation within this diagnostic classification. PMID- 19903733 TI - Effects of baclofen on motor units paralysed by chronic cervical spinal cord injury. AB - Baclofen, a gamma-aminobutyric acid receptor(B) agonist, is used to reduce symptoms of spasticity (hyperreflexia, increases in muscle tone, involuntary muscle activity), but the long-term effects of sustained baclofen use on skeletal muscle properties are unclear. The aim of our study was to evaluate whether baclofen use and paralysis due to cervical spinal cord injury change the contractile properties of human thenar motor units more than paralysis alone. Evoked electromyographic activity and force were recorded in response to intraneural stimulation of single motor axons to thenar motor units. Data from three groups of motor units were compared: 23 paralysed units from spinal cord injured subjects who take baclofen and have done so for a median of 7 years, 25 paralysed units from spinal cord injured subjects who do not take baclofen (median: 10 years) and 45 units from uninjured control subjects. Paralysed motor unit properties were independent of injury duration and level. With paralysis and baclofen, the median motor unit tetanic forces were significantly weaker, twitch half-relaxation times longer and half maximal forces reached at lower frequencies than for units from uninjured subjects. The median values for these same parameters after paralysis alone were comparable to control data. Axon conduction velocities differed across groups and were slowest for paralysed units from subjects who were not taking baclofen and fastest for units from the uninjured. Greater motor unit weakness with long-term baclofen use and paralysis will make the whole muscle weaker and more fatigable. Significantly more paralysed motor units need to be excited during patterned electrical stimulation to produce any given force over time. The short-term benefits of baclofen on spasticity (e.g. management of muscle spasms that may otherwise hinder movement or social interactions) therefore have to be considered in relation to its possible long term effects on muscle rehabilitation. Restoring the strength and speed of paralysed muscles to pre-injury levels may require more extensive therapy when baclofen is used chronically. PMID- 19903735 TI - Protein disulphide isomerase protects against protein aggregation and is S nitrosylated in amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis is a rapidly progressing fatal neurodegenerative disease characterized by the presence of protein inclusions within affected motor neurons. Endoplasmic reticulum stress leading to apoptosis was recently recognized to be an important process in the pathogenesis of sporadic human amyotrophic lateral sclerosis as well as in transgenic models of mutant superoxide dismutase 1-linked familial amyotrophic lateral sclerosis. Endoplasmic reticulum stress occurs early in disease, indicating a critical role in pathogenesis, and involves upregulation of an important endoplasmic reticulum chaperone, protein disulphide isomerase. We aimed to investigate the involvement of protein disulphide isomerase in endoplasmic reticulum stress induction, protein aggregation, inclusion formation and toxicity in amyotrophic lateral sclerosis. Motor neuron-like NSC-34 cell lines were transfected with superoxide dismutase 1 and protein disulphide isomerase encoding vectors and small interfering RNA, and examined by immunocytochemistry and immunoblotting. Expression of mutant superoxide dismutase 1 induced endoplasmic reticulum stress, predominantly in cells bearing mutant superoxide dismutase 1 inclusions but also in a proportion of cells expressing mutant superoxide dismutase 1 without visible inclusions. Over-expression of protein disulphide isomerase decreased mutant superoxide dismutase 1 aggregation, inclusion formation, endoplasmic reticulum stress induction and toxicity, whereas small interfering RNA targeting protein disulphide isomerase increased mutant superoxide dismutase 1 inclusion formation, indicating a protective role for protein disulphide isomerase against superoxide dismutase 1 misfolding. Aberrant modification of protein disulphide isomerase by S-nitrosylation of active site cysteine residues has previously been shown as an important process in neurodegeneration in Parkinson's and Alzheimer's disease brain tissue, but has not been described in amyotrophic lateral sclerosis. Using a biotin switch assay, we detected increased levels of S-nitrosylated protein disulphide isomerase in transgenic mutant superoxide dismutase 1 mouse and human sporadic amyotrophic lateral sclerosis spinal cord tissues. Hence, despite upregulation, protein disulphide isomerase is also functionally inactivated in amyotrophic lateral sclerosis, which may prevent its normal protective function and contribute to disease. We also found that a small molecule mimic of the protein disulphide isomerase active site, (+/-)-trans-1,2 bis(mercaptoacetamido)cyclohexane, protected against mutant superoxide dismutase 1 inclusion formation. These studies reveal that endoplasmic reticulum stress is important in the formation of mutant superoxide dismutase 1 inclusions, and protein disulphide isomerase has an important function in ameliorating mutant superoxide dismutase 1 aggregation and toxicity. Functional inhibition of protein disulphide isomerase by S-nitrosylation may contribute to pathophysiology in both mutant superoxide dismutase 1-linked disease and sporadic amyotrophic lateral sclerosis. Protein disulphide isomerase is therefore a novel potential therapeutic target in amyotrophic lateral sclerosis and (+/-)-trans-1,2 bis(mercaptoacetamido)cyclohexane and other molecular mimics of protein disulphide isomerase could be of benefit in amyotrophic lateral sclerosis and other neurodegenerative diseases related to protein misfolding. PMID- 19903736 TI - Anterior temporal lobe connectivity correlates with functional outcome after aphasic stroke. AB - Focal brain lesions are assumed to produce language deficits by two basic mechanisms: local cortical dysfunction at the lesion site, and remote cortical dysfunction due to disruption of the transfer and integration of information between connected brain regions. However, functional imaging studies investigating language outcome after aphasic stroke have tended to focus only on the role of local cortical function. In this positron emission tomography functional imaging study, we explored relationships between language comprehension performance after aphasic stroke and the functional connectivity of a key speech-processing region in left anterolateral superior temporal cortex. We compared the organization of left anterolateral superior temporal cortex functional connections during narrative speech comprehension in normal subjects with left anterolateral superior temporal cortex connectivity in a group of chronic aphasic stroke patients. We then evaluated the language deficits associated with altered left anterolateral superior temporal cortex connectivity in aphasic stroke. During normal narrative speech comprehension, left anterolateral superior temporal cortex displayed positive functional connections with left anterior basal temporal cortex, left inferior frontal gyrus and homotopic cortex in right anterolateral superior temporal cortex. As a group, aphasic patients demonstrated a selective disruption of the normal functional connection between left and right anterolateral superior temporal cortices. We observed that deficits in auditory single word and sentence comprehension correlated both with the degree of disruption of left-right anterolateral superior temporal cortical connectivity and with local activation in the anterolateral superior temporal cortex. Subgroup analysis revealed that aphasic patients with preserved positive intertemporal connectivity displayed better receptive language function; these patients also showed greater than normal left inferior frontal gyrus activity, suggesting a possible 'top-down' compensatory mechanism. These results demonstrate that functional connectivity between anterolateral superior temporal cortex and right anterior superior temporal cortex is a marker of receptive language outcome after aphasic stroke, and illustrate that language system organization after focal brain lesions may be marked by complex signatures of altered local and pathway-level function. PMID- 19903737 TI - Smoking cessation for hospitalized smokers: an evaluation of the "Ottawa Model". AB - INTRODUCTION: Interventions for hospitalized smokers can increase long-term smoking cessation rates. The Ottawa Model for Smoking Cessation (the "Ottawa Model") is an application of the "5 A's" approach to cessation, customized to the hospital setting. This study evaluated the impact of implementing the Ottawa Model in 9 hospitals in eastern Ontario. METHODS: The RE-AIM (Reach, Efficacy, Adoption, Implementation, and Maintenance) framework was used to evaluate the intervention. Trained outreach facilitators assisted 9 hospitals to implement the Ottawa Model; program delivery was then monitored over a 1-year period using administrative data and data from a follow-up database. A before-and-after study was conducted to gauge the effect of the Ottawa Model program on cessation rates 6 months after hospitalization. Self-reports of smoking cessation were biochemically confirmed in a random sample of patients, and all cessation rates were corrected for potential misreporting. RESULTS: Sixty-nine percent of the expected number of smokers received the Ottawa Model intervention. Controlling for hospital, the confirmed 6-month continuous abstinence rate was higher after, than before, introduction of the Ottawa Model (29.4% vs. 18.3%; odds ratio = 1.71, 95% CI = 1.11-2.64; Z = 2.43; I(2) = 0%; p = .02). The intervention was more likely to accomplish counseling for smokers than delivery of medications or postdischarge follow-up. Attitudinal, managerial, and environmental challenges to program implementation were identified. DISCUSSION: Trained outreach facilitators successfully implemented the Ottawa Model in 9 hospitals leading to significantly higher long-term cessation rates. The public health implications of systematic cessation programs for hospitalized smokers are profound. PMID- 19903738 TI - Bacterial display enables efficient and quantitative peptide affinity maturation. AB - A quantitative screening method was developed to enable isolation and affinity maturation of peptide ligands specific for a given target from peptide libraries displayed on the outer surface of Escherichia coli using multi-parameter flow cytometry. From a large, random 15-mer peptide library, screening identified a core motif of W-E/D-W-E/D that conferred binding to vascular endothelial growth factor (VEGF). One cycle of affinity maturation resulted in the identification of several families of VEGF-binding peptides having distinct consensus sequences, from which a preferred disulfide constraint emerged. In the second affinity maturation cycle, high affinity peptides were favored by the addition of a decoy protein that bound an adjacent epitope on the display scaffold. The decoy apparently reduced rebinding or avidity effects, and the resulting peptides exhibited consensus at 12 of 19 amino acid positions. Peptides identified and affinity matured using bacterial display were remarkably similar to the best affinity matured using phage display and exhibited comparable dissociation constants (within 2-fold; K(D) = 4.7 x 10(-7) M). Screening of bacterial displayed peptide libraries using cytometry enabled optimization of screening conditions to favor affinity and specificity and rapid clonal characterization. Bacterial display thus provides a new quantitative tool for the discovery and evolutionary optimization of protein-specific peptide ligands. PMID- 19903739 TI - Molecular and metabolic evidence for mitochondrial defects associated with beta cell dysfunction in a mouse model of type 2 diabetes. AB - OBJECTIVE The inability of pancreatic beta-cells to appropriately respond to glucose and secrete insulin are primary defects associated with beta-cell failure in type 2 diabetes. Mitochondrial dysfunction has been implicated as a key factor in the development of type 2 diabetes; however, a link between mitochondrial dysfunction and defective insulin secretion is unclear. RESEARCH DESIGN AND METHODS We investigated the changes in islet mitochondrial function and morphology during progression from insulin resistance (3 weeks old), immediately before hyperglycemia (5 weeks old), and after diabetes onset (10 weeks old) in transgenic MKR mice compared with controls. The molecular and protein changes at 10 weeks were determined using microarray and iTRAQ proteomic screens. RESULTS At 3 weeks, MKR mice were hyperinsulinemic but normoglycemic and beta-cells showed negligible mitochondrial or morphological changes. At 5 weeks, MKR islets displayed abrogated hyperpolarization of mitochondrial membrane potential (DeltaPsi(m)), reduced mitochondrial Ca(2+) uptake, slightly enlarged mitochondria, and reduced glucose-stimulated insulin secretion. By 10 weeks, MKR mice were hyperglycemic and hyperinsulinemic and beta-cells contained swollen mitochondria with disordered cristae. beta-Cells displayed impaired stimulus secretion coupling including reduced hyperpolarization of DeltaPsi(m), impaired Ca(2+)-signaling, and reduced glucose-stimulated ATP/ADP and insulin release. Furthermore, decreased cytochrome c oxidase-dependent oxygen consumption and signs of oxidative stress were observed in diabetic islets. Protein profiling of diabetic islets revealed that 36 mitochondrial proteins were differentially expressed, including inner membrane proteins of the electron transport chain. CONCLUSIONS We provide novel evidence for a critical role of defective mitochondrial oxidative phosphorylation and morphology in the pathology of insulin resistance-induced beta-cell failure. PMID- 19903740 TI - Invariant natural killer T-cell control of type 1 diabetes: a dendritic cell genetic decision of a silver bullet or Russian roulette. AB - OBJECTIVE: In part, activation of invariant natural killer T (iNKT)-cells with the superagonist alpha-galactosylceramide (alpha-GalCer) inhibits the development of T-cell-mediated autoimmune type 1 diabetes in NOD mice by inducing the downstream differentiation of antigen-presenting dendritic cells (DCs) to an immunotolerogenic state. However, in other systems iNKT-cell activation has an adjuvant-like effect that enhances rather than suppresses various immunological responses. Thus, we tested whether in some circumstances genetic variation would enable activated iNKT-cells to support rather than inhibit type 1 diabetes development. RESEARCH DESIGN AND METHODS: We tested whether iNKT-conditioned DCs in NOD mice and a major histocompatibility complex-matched C57BL/6 (B6) background congenic stock differed in capacity to inhibit type 1 diabetes induced by the adoptive transfer of pathogenic AI4 CD8 T-cells. RESULTS: Unlike those of NOD origin, iNKT-conditioned DCs in the B6 background stock matured to a state that actually supported rather than inhibited AI4 T-cell-induced type 1 diabetes. The induction of a differing activity pattern of T-cell costimulatory molecules varying in capacity to override programmed death-ligand-1 inhibitory effects contributes to the respective ability of iNKT-conditioned DCs in NOD and B6 background mice to inhibit or support type 1 diabetes development. Genetic differences inherent to both iNKT-cells and DCs contribute to their varying interactions in NOD and B6.H2(g7) mice. CONCLUSIONS: This great variability in the interactions between iNKT-cells and DCs in two inbred mouse strains should raise a cautionary note about considering manipulation of this axis as a potential type 1 diabetes prevention therapy in genetically heterogeneous humans. PMID- 19903741 TI - Positive regulation of spondin 2 by thyroid hormone is associated with cell migration and invasion. AB - The thyroid hormone 3,3',5-triiodo-L-thyronine (T(3)) regulates growth, development, and differentiation processes in animals. These activities are mediated by the nuclear thyroid hormone receptors (TRs). Microarray analyses were performed previously to study the mechanism of regulation triggered by T(3) treatment in hepatoma cell lines. The results showed that spondin 2 was regulated positively by T(3). However, the underlying mechanism and the physiological role of T(3) in the regulation of spondin 2 are not clear. To verify the microarray results, spondin 2 was further investigated using semi-quantitative reverse transcription-PCR and western blotting. After 48 h of T(3) treatment in the HepG2 TR alpha 1#1 cell line, spondin 2 mRNA and protein levels increased by 3.9- to 5.7-fold. Similar results were observed in thyroidectomized rats. To localize the regulatory region in spondin 2, we performed serial deletions of the promoter and chromatin immunoprecipitation assays. The T(3) response element on the spondin 2 promoter was localized in the -1104/-1034 or -984/-925 regions. To explore the effect of spondin 2 on cellular function, spondin 2 knockdown cell lines were established from Huh7 cells. Knockdown cells had higher migration ability and invasiveness compared with control cells. Conversely, spondin 2 overexpression in J7 cells led to lower migration ability and invasiveness compared with control cells. Furthermore, this study demonstrated that spondin 2 overexpression in some types of hepatocellular carcinomas is TR dependent. Together, these experimental findings suggest that spondin 2, which is regulated by T(3), has an important role in cell invasion, cell migration, and tumor progression. PMID- 19903742 TI - The Ca2+-calmodulin-dependent kinase II is activated in papillary thyroid carcinoma (PTC) and mediates cell proliferation stimulated by RET/PTC. AB - RET/papillary thyroid carcinoma (PTC), TRK-T, or activating mutations of Ras and BRaf are frequent genetic alterations in PTC, all leading to the activation of the extracellular-regulated kinase (Erk) cascade. The aim of this study was to investigate the role of calmodulin-dependent kinase II (CaMKII) in the signal transduction leading to Erk activation in PTC cells. In normal thyroid cells, CaMKII and Erk were in the inactive form in the absence of stimulation. In primary PTC cultures and in PTC cell lines harboring the oncogenes RET/PTC-1 or BRaf(V600E), CaMKII was active also in the absence of any stimulation. Inhibition of calmodulin or phospholipase C (PLC) attenuated the level of CaMKII activation. Expression of recombinant RET/PTC-3, BRaf(V600E), or Ras(V12) induced CaMKII activation. Inhibition of CaMKII attenuated Erk activation and DNA synthesis in thyroid papillary carcinoma (TPC-1), a cell line harboring RET/PTC-1, suggesting that CaMKII is a component of the Erk signal cascade in this cell line. In conclusion, PTCs contain an active PLC/Ca(2+)/calmodulin-dependent signal inducing constitutive activation of CaMKII. This kinase is activated by BRaf(V600E), oncogenic Ras, and by RET/PTC. CaMKII participates to the activation of the Erk pathway by oncogenic Ras and RET/PTC and contributes to their signal output, thus modulating tumor cell proliferation. PMID- 19903743 TI - MicroRNAs in ovarian carcinomas. AB - The molecular mechanisms involved in epithelial ovarian cancer initiation and progression are just beginning to be elucidated. In particular, it has become evident that microRNAs (miRNAs or miRs), a class of molecules that post transcriptionally regulate gene expression, play a major role in ovarian tumorigenesis. Several microRNA profiling studies have identified changes in microRNA patterns that take place during ovarian cancer development. While most deregulated microRNAs are down-regulated in cancer, and may therefore act as tumor suppressors, others are elevated and may represent novel oncogenes in this disease. A number of microRNAs identified as aberrantly expressed in ovarian carcinoma have been shown to have important functional roles in cancer development and may therefore represent targets for therapy. In addition, some of the microRNA patterns may have prognostic significance. The identification of functional targets represents a major hurdle in our understanding of microRNA function in ovarian carcinoma, but significant progress is being made. It is hoped that a better understanding of the microRNA expression and roles in ovarian cancer may provide new avenues for the detection, diagnosis, and therapy of this deadly disease. PMID- 19903744 TI - Plasma sex hormone concentrations and breast cancer risk in an ethnically diverse population of postmenopausal women: the Multiethnic Cohort Study. AB - To add to the existing evidence that comes mostly from White populations, we conducted a nested case-control study to examine the association between sex hormones and breast cancer risk within the Multiethnic Cohort that includes Japanese American, White, Native Hawaiian, African American, and Latina women. Of the postmenopausal women for whom we had a plasma sample, 132 developed breast cancer during follow-up. Two controls per case, matched on study area (Hawaii, Los Angeles), ethnicity/race, birth year, date and time of blood draw and time fasting, were randomly selected from the women who had not developed breast cancer. Levels of estradiol (E(2)), estrone (E(1)), androstenedione, dehydroepiandrosterone (DHEA), and testosterone were quantified by RIA after organic extraction and Celite column partition chromatography. E(1) sulfate, DHEA sulfate (DHEAS), and sex hormone-binding globulin (SHBG) were quantified by direct immunoassays. Based on conditional logistic regression, the sex hormones were positively associated and SHBG was negatively associated with breast cancer risk. All associations, except those with DHEAS and testosterone showed a significant linear trend. The odds ratio (OR) associated with a doubling of E(2) levels was 2.26 (95% confidence interval (CI) 1.58-3.25), and the OR associated with a doubling of testosterone levels was 1.34 (95% CI 0.98-1.82). The associations in Japanese American women, who constituted 54% of our sample, were similar to or nonsignificantly stronger than in the overall group. This study provides the best evidence to date that the association between sex hormones and breast cancer risk is generalizable to an ethnically diverse population. PMID- 19903745 TI - The clusterin paradigm in prostate and breast carcinogenesis. AB - The biological functions of clusterin (CLU, also known as ApoJ, SGP2, TRPM-2, CLI) have been puzzling the researchers since its first discovery in the early 80's. We know that CLU is a single 9-exons gene expressing three protein forms with different sub-cellular localisations and diverse biological functions. Despite the many reports from many research teams on CLU action and its relation to tumourigenesis, contradictions in the data and alternative hypothesis still exist. Understanding the role of CLU in tumourigenesis is complicated not only by the existence of different protein forms but also by the changes of tumours over time and the selection pressures imposed by treatments such as hormone ablation or chemotherapy. This review focuses on recent discoveries concerning the role of CLU in prostate and breast cancer onset and progression. Although CLU acts primarily as a tumour suppressor in the early stages of carcinogenesis, consistent with its role in the involution of the prostate following castration, late stage cancer may overexpress CLU following chemotherapeutic drugs or hormonal ablation therapy. High expression of secreted or cytoplasmic CLU may represent a pro-survival stimulus because it confers increased resistance to killing by anti-cancer drugs or enhances tumour cell survival in specific niches. PMID- 19903747 TI - The dawn of developmental signaling in the metazoa. AB - Intercellular signaling underpins metazoan development by mediating the induction, organization, and cooperation of cells, tissues, and organs. Herein, the origins of the four major signaling pathways used during animal development and differentiation-Wnt, Notch, transforming growth factor-beta (TGF-beta), and Hedgehog-are assessed by comparative analysis of genomes from bilaterians, early branching metazoan phyla (poriferans, placozoans, and cnidarians), and the holozoan sister clade to the animal kingdom, the choanoflagellates. On the basis of the incidence and domain architectures of core pathway ligands, receptors, signal transducers, and transcription factors in representative species of these lineages, it appears that the Notch, Wnt, and TGF-beta pathways are metazoan synapomorphies, whereas the Hedgehog pathway arose in the protoeumetazoan lineage, after its divergence from poriferan and placozoan lineages. Examination of the binding domains and motifs present in signaling pathway components of nonbilaterians reveals cases in which signaling interactions are unlikely to be operating in accordance with bilaterian canons. Overall, this study highlights the stability and antiquity of the core cytosolic components of each pathway, juxtaposed with the more variable and recently evolved molecular interactions taking place at the cell surface. PMID- 19903746 TI - Review of pathological hallmarks of schizophrenia: comparison of genetic models with patients and nongenetic models. AB - Schizophrenia is a condition that impairs higher brain functions, some of which are specific to humans. After identification of susceptibility genes for schizophrenia, many efforts have been made to generate genetics-based models for the disease. It is under debate whether behavioral deficits observed in rodents are sufficient to characterize these models. Alternatively, anatomical and neuropathological changes identified in brains of patients with schizophrenia may be utilized as translatable characteristics between humans and rodents, which are important for validation of the models. Here, we overview such anatomical and neuropathological changes in humans: enlarged ventricles, dendritic changes in the pyramidal neurons, and alteration of specific subtypes of interneurons. In this review, we will overview such morphological changes in brains from patients with schizophrenia. Then, we will describe that some of these alterations are already recapitulated even in classic nongenetic models for schizophrenia. Finally, in comparison with the changes in patients and nongenetic models, we will discuss the anatomical and neuropathological manifestation in genetic models for schizophrenia. PMID- 19903748 TI - Why sex and recombination? AB - Sex and recombination have long been seen as adaptations that facilitate natural selection by generating favorable variations. If recombination is to aid selection, there must be negative linkage disequilibria-favorable alleles must be found together less often than expected by chance. These negative linkage disequilibria can be generated directly by selection, but this must involve negative epistasis of just the right strength, which is not expected, from either experiment or theory. Random drift provides a more general source of negative associations: Favorable mutations almost always arise on different genomes, and negative associations tend to persist, precisely because they shield variation from selection. We can understand how recombination aids adaptation by determining the maximum possible rate of adaptation. With unlinked loci, this rate increases only logarithmically with the influx of favorable mutations. With a linear genome, a scaling argument shows that in a large population, the rate of adaptive substitution depends only on the expected rate in the absence of interference, divided by the total rate of recombination. A two-locus approximation predicts an upper bound on the rate of substitution, proportional to recombination rate. If associations between linked loci do impede adaptation, there can be substantial selection for modifiers that increase recombination. Whether this can account for the maintenance of high rates of sex and recombination depends on the extent of selection. It is clear that the rate of species-wide substitutions is typically far too low to generate appreciable selection for recombination. However, local sweeps within a subdivided population may be effective. PMID- 19903749 TI - Selection, gene interaction, and flexible gene networks. AB - Recent results from a variety of different kinds of experiments, mainly using behavior as an assay, and ranging from laboratory selection experiments to gene interaction studies, show that a much wider range of genes can affect phenotype than those identified as "core genes" in classical mutant screens. Moreover, very pleiotropic genes can produce specific phenotypes when mild variants are combined. These studies also show that gene networks readily change configuration and the relationships between interacting genes in response to the introduction of additional genetic variants, suggesting that the networks range widely and have a high degree of flexibility and malleability. Such flexibility, in turn, offers a plausible mechanism for the molding of phenotypes through microevolution, as a prerequisite to making a suitable environment for the acceptance of newly arising large-effect mutations in the transition from microevolution to macroevolution. PMID- 19903750 TI - Evolving views of DNA replication (in)fidelity. AB - "It has not escaped our notice that the specific pairing we have postulated immediately suggests a possible copying mechanism for the genetic material" (Watson and Crick 1953). In the years since this remarkable understatement, we have come to realize the enormous complexity of the cellular machinery devoted to replicating DNA with the accuracy needed to maintain genetic information over many generations, balanced by the emergence of mutations on which selection can act. This complexity is partly based on the need to remove or tolerate cytotoxic and mutagenic lesions in DNA generated by environmental stress. Considered here is the fidelity with which undamaged and damaged DNA is replicated by the many DNA polymerases now known to exist. Some of these seriously violate Watson-Crick base-pairing rules such that, depending on the polymerase, the composition and location of the error, and the ability to correct errors (or not), DNA synthesis error rates can vary by more than a millionfold. This offers the potential to modulate rates of point mutations over a wide range, with consequences that can be either deleterious or beneficial. PMID- 19903751 TI - Familial risks for type 2 diabetes in Sweden. AB - OBJECTIVE: Our aim was to characterize familial risks for type 2 diabetes by the type and number of affected family members, including half-siblings, adoptees, and spouses, to quantify risks and estimate the contribution of environmental effect. RESEARCH DESIGN AND METHODS: Families were identified from the Multigeneration Register, and type 2 diabetic patients were obtained from the Hospital Discharge Register. Standardized incidence ratios were calculated for offspring with type 2 diabetes whose family members were hospitalized for type 2 diabetes at ages >39 years compared with those lacking affected family members. RESULTS: The number of hospitalized type 2 diabetic patients was 157,549. Among 27,895 offspring, 27.9% had a parent or sibling also hospitalized for type 2 diabetes. The familial relative risk (RR) ranged from 2.0 to >30, depending on the number and type of probands. The highest RRs of type 2 diabetes were found in individuals who had at least two siblings affected by type 2 diabetes, irrespective of the parental disease. Adoptees showed no risk from adopted parents. CONCLUSIONS: The study, the largest yet published, showed that familial RRs varied by the number and type of affected family member. However, much of the familial clustering remains yet to be genetically explained. The high risk should be recognized in clinical genetic counseling. The data from adoptees confirmed the genetic basis of the familial associations, but those from half siblings and spouses suggested that a smaller part of familial clustering may be accounted for by environmental factors. PMID- 19903752 TI - A1C cut points to define various glucose intolerance groups in Asian Indians. AB - OBJECTIVE To determine A1C cut points for glucose intolerance in Asian Indians. RESEARCH DESIGN AND METHODS A total of 2,188 participants without known diabetes were randomly selected from the Chennai Urban Rural Epidemiology Study. All had fasting plasma glucose (FPG) and 2-h postload plasma glucose measurements after a 75-g load and were classified as having impaired fasting glucose (IFG) (American Diabetes Association [ADA] criteria, FPG > or =5.5 and <7 mmol/l, and World Health Organization [WHO] criteria, FPG > or =6.1 and <7 mmol/l), impaired glucose tolerance (IGT) (2-h postload plasma glucose > or =7.8 and <11.1 mmol/l), or diabetes (FPG > or =7 mmol/l and/or 2-h postload plasma glucose > or =11.1 mmol/l). A1C was measured using the Bio-Rad Variant machine. Based on receiver operating characteristic curves, optimum sensitivity and specificity were derived for defining A1C cut points for diabetes, IGT, and IFG. RESULTS Mean +/- SD values of A1C among subjects with normal glucose tolerance, IGT, and diabetes were 5.5 +/- 0.4, 5.9 +/- 0.6, and 8.3 +/- 2.0%, respectively (P(trend) < 0.001) with considerable overlap. To identify diabetes based on 2-h postload plasma glucose, the A1C cut point of 6.1% had an area under the curve (AUC) of 0.941 with 88.0% sensitivity and 87.9% specificity. When diabetes was defined as FPG > or =7.0 mmol/l, the A1C cut point was 6.4% (AUC = 0.966, sensitivity 93.3%, and specificity 92.3%). For IGT, AUC = 0.708; for IFG, AUC = 0.632 (WHO criteria) and 0.708 (ADA criteria), and the A1C cut point was 5.6%. CONCLUSIONS In Asian Indians, A1C cut points of 6.1 and 6.4% defined diabetes by 2-h postload plasma glucose or FPG criteria, respectively. A value of 5.6% optimally identified IGT or IFG but was <70% accurate. PMID- 19903753 TI - Epidemiological perspectives on type 1 diabetes in childhood and adolescence in germany: 20 years of the Baden-wurttemberg Diabetes Incidence Registry (DIARY). AB - OBJECTIVE: To predict the frequency of type 1 diabetes in childhood and adolescence (<15 years of age) in Germany for the next 20 years. RESEARCH DESIGN AND METHODS: Data on diabetes onset has been collected by means of a registry in the federal German state of Baden-Wurttemberg (documentation period, 1987-2006; n = 5,108; completeness of data 98.1%). RESULTS: The current incidence rate (2000 2006) is 19.4 per 100,000 per year (95% CI 18.6-20.2). The annual incidence rate can be expressed as a square of a linear function of the calendar year X [y = (3.05 + 0.0778 x {X-1986})(2); r(2) = 0.90]. The highest increase per year was observed in the age-groups comprising 2- and 3-year-olds (12 and 13% per year, respectively). The incidence rate for the year 2026 is estimated to be 37.9 per 100,000 per year (95% CI 33.3-42.9). CONCLUSIONS: The increase that we found in younger children is characteristic of a left shift toward an earlier age. PMID- 19903754 TI - Clinical heterogeneity in monogenic diabetes caused by mutations in the glucokinase gene (GCK-MODY). AB - OBJECTIVE: To evaluate the heterogeneity in the clinical expression in a family with glucokinase mature-onset diabetes of the young (GCK-MODY). RESEARCH DESIGN AND METHODS: Members (three generations) of the same family presented either with overt neonatal hyperglycemia, marked postprandial hyperglycemia, or glucosuria. Homeostasis model assessment of insulin resistance (HOMA(IR)) and insulinogenic and disposition indexes were calculated. Oral glucose tolerance test (OGTT) results in the GCK mutation carriers from this family were compared with those from other subjects with GCK mutations in the same codon (GCK(261)), with other missense and other types of GCK mutations in different codons from the European MODY Consortium database (GCK(m)). RESULTS: Mutation G261R was found in the GCK gene. During the OGTT, glucose (P = 0.02) and insulin (P = 0.009) response at 2 h as well as at the 2-h glucose increment (GCK(261) versus other missense GCK mutations, P = 0.003) were significantly higher in GCK(261) than in GCK(m) carriers. CONCLUSIONS: Differing from other GCK(m) carriers, the glucose and insulin response to oral glucose was significantly higher in GCK(261) carriers, indicating clinical heterogeneity in GCK-MODY. PMID- 19903756 TI - Effectiveness of acute medical units in hospitals: a systematic review. AB - PURPOSE: To assess the effectiveness of acute medical units (AMUs) in hospitals. DATA SOURCES: (i) Controlled and observational studies in peer-reviewed journals retrieved from PubMed, EPOC, CINAHL and ERIC databases published between January 1990 and July 2008; and (ii) reports from non-peer-reviewed websites combined with Google search. STUDY SELECTION: Articles reporting effects of the introduction of an AMU on mortality, length of stay, discharge disposition, readmissions, resource use and patient and/or staff satisfaction. Data extraction Data on unit operations and outcome measures were extracted by a single author and confirmed by a second author, with disagreement settled by consensus. RESULTS OF DATA SYNTHESIS: Nine peer-reviewed reports of before-after analyses of seven units introduced into the UK and Ireland were analysed. Two studies, one prospective, reported significant reductions in in-patient mortality between 0.6 and 5.6% points following commencement of AMU. Four studies reported significant reductions in the length of stay between 1.5 and 2.5 days. Waiting times for patient transfer from emergency departments to medical beds decreased by 30% in one study. In three studies, the proportion of medical patients discharged directly home from the AMU increased by 8-25% points. Three studies noted no increase in 30-day readmission rates following unit commencement. Two studies described significant improvements in patient and staff satisfaction with care. Eight non-peer-reviewed reports relating to 48 units confirmed reductions in the length of stay. CONCLUSION: Limited observational data suggest AMUs reduce in patient mortality, length of stay and emergency department access block without increasing readmission rates, and improve patient and staff satisfaction. PMID- 19903755 TI - Relations of dietary magnesium intake to biomarkers of inflammation and endothelial dysfunction in an ethnically diverse cohort of postmenopausal women. AB - OBJECTIVE: Although magnesium may favorably affect metabolic outcomes, few studies have investigated the role of magnesium intake in systemic inflammation and endothelial dysfunction in humans. RESEARCH DESIGN AND METHODS: Among 3,713 postmenopausal women aged 50-79 years in the Women's Health Initiative Observational Study and free of cardiovascular disease, cancer, and diabetes at baseline, we measured plasma concentrations of high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), turnor necrosis factor-alpha receptor 2 (TNF-alpha-R2), soluble intercellular adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), and E-selectin. Magnesium intake was assessed using a semiquantitative food frequency questionnaire. RESULTS: After adjustment for age, ethnicity, clinical center, time of blood draw, smoking, alcohol, physical activity, energy intake, BMI, and diabetes status, magnesium intake was inversely associated with hs-CRP (P for linear trend = 0.003), IL-6 (P < 0.0001), TNF-alpha-R2 (P = 0.0006), and sVCAM-1 (P = 0.06). Similar findings remained after further adjustment for dietary fiber, fruit, vegetables, folate, and saturated and trans fat intake. Multivariable-adjusted geometric means across increasing quintiles of magnesium intake were 3.08, 2.63, 2.31, 2.53, and 2.16 mg/l for hs-CRP (P = 0.005); 2.91, 2.63, 2.45, 2.27, and 2.26 pg/ml for IL-6 (P = 0.0005); and 707, 681, 673, 671, and 656 ng/ml for sVCAM-1 (P = 0.04). An increase of 100 mg/day magnesium was inversely associated with hs-CRP (-0.23 mg/l +/- 0.07; P = 0.002), IL-6 (-0.14 +/- 0.05 pg/ml; P = 0.004), TNF-alpha-R2 (-0.04 +/- 0.02 pg/ml; P = 0.06), and sVCAM-1 (-0.04 +/- 0.02 ng/ml; P = 0.07). No significant ethnic differences were observed. CONCLUSIONS: High magnesium intake is associated with lower concentrations of certain markers of systemic inflammation and endothelial dysfunction in postmenopausal women. PMID- 19903757 TI - Determination of health-care teamwork training competencies: a Delphi study. AB - OBJECTIVE: The purpose of this study was to determine the optimum content of a 1 day classroom-based crew resource management (CRM) course for health-care personnel working in ad hoc teams in complex, time-critical hospital departments such as surgery, intensive care or emergency. DESIGN: A two-round modified Delphi panel. PARTICIPANTS: selected teamwork competency components suitable for inclusion in 1 day of training from a list developed via literature review. Participants Fifteen experts in health care, CRM and training. MAIN OUTCOME MEASURE: Knowledge, skill and attitude competency components for a 1-day CRM course. RESULTS: Of the 110 knowledge, skill and attitude CRM competency components, 40 components were selected by greater than 70% of respondents, whereas the remaining 62 components were selected by fewer than 55% of respondents. These 40 competency components ranged across five competency domains: communication, task management, situational awareness, decision-making and leadership, and provided a consensus on the most critical areas for inclusion in training for health-care personnel. CONCLUSIONS: This new competency model is now available for use. Although the sample size was limited, a high degree of consensus was reached after only two rounds. A modified Delphi technique within the context of competencies first refined from the literature was a useful and cost-effective method for determining the content of a 1-day CRM training course for health-care workers. PMID- 19903758 TI - A conserved transcriptional regulator is required for RNA-directed DNA methylation and plant development. AB - RNA-directed DNA methylation (RdDM) is a conserved mechanism for epigenetic silencing of transposons and other repetitive elements. We report that the rdm4 (RNA-directed DNA Methylation4) mutation not only impairs RdDM, but also causes pleiotropic developmental defects in Arabidopsis. Both RNA polymerase II (Pol II) and Pol V-dependent transcripts are affected in the rdm4 mutant. RDM4 encodes a novel protein that is conserved from yeast to humans and interacts with Pol II and Pol V in plants. Our results suggest that RDM4 functions in epigenetic regulation and plant development by serving as a transcriptional regulator for RNA Pol V and Pol II, respectively. PMID- 19903759 TI - Dicer1 functions as a haploinsufficient tumor suppressor. AB - While the global down-regulation of microRNAs (miRNAs) is a common feature of human tumors, its genetic basis is largely undefined. To explore this question, we analyzed the consequences of conditional Dicer1 mutation (Dicer1 "floxed" or Dicer1(fl)) on several mouse models of cancer. Here we show Dicer1 functions as a haploinsufficient tumor suppressor gene. Deletion of a single copy of Dicer1 in tumors from Dicer1(fl/+) animals led to reduced survival compared with controls. These tumors exhibited impaired miRNA processing but failed to lose the wild-type Dicer1 allele. Moreover, tumors from Dicer1(fl/fl) animals always maintained one functional Dicer1 allele. Consistent with selection against full loss of Dicer1 expression, enforced Dicer1 deletion caused inhibition of tumorigenesis. Analysis of human cancer genome copy number data reveals frequent deletion of DICER1. Importantly, however, the gene has not been reported to undergo homozygous deletion, suggesting that DICER1 is haploinsufficient in human cancer. These findings suggest Dicer1 may be an important haploinsufficient tumor suppressor gene and, furthermore, that other factors controlling miRNA biogenesis may also function in this manner. PMID- 19903760 TI - Inferring tumor progression from genomic heterogeneity. AB - Cancer progression in humans is difficult to infer because we do not routinely sample patients at multiple stages of their disease. However, heterogeneous breast tumors provide a unique opportunity to study human tumor progression because they still contain evidence of early and intermediate subpopulations in the form of the phylogenetic relationships. We have developed a method we call Sector-Ploidy-Profiling (SPP) to study the clonal composition of breast tumors. SPP involves macro-dissecting tumors, flow-sorting genomic subpopulations by DNA content, and profiling genomes using comparative genomic hybridization (CGH). Breast carcinomas display two classes of genomic structural variation: (1) monogenomic and (2) polygenomic. Monogenomic tumors appear to contain a single major clonal subpopulation with a highly stable chromosome structure. Polygenomic tumors contain multiple clonal tumor subpopulations, which may occupy the same sectors, or separate anatomic locations. In polygenomic tumors, we show that heterogeneity can be ascribed to a few clonal subpopulations, rather than a series of gradual intermediates. By comparing multiple subpopulations from different anatomic locations, we have inferred pathways of cancer progression and the organization of tumor growth. PMID- 19903761 TI - Differential genomic responses in old vs. young humans despite similar levels of modest muscle damage after resistance loading. AB - Across numerous model systems, aging skeletal muscle demonstrates an impaired regenerative response when exposed to the same stimulus as young muscle. To better understand the impact of aging in a human model, we compared changes to the skeletal muscle transcriptome induced by unaccustomed high-intensity resistance loading (RL) sufficient to cause moderate muscle damage in young (37 yr) vs. older (73 yr) adults. Serum creatine kinase was elevated 46% 24 h after RL in all subjects with no age differences, indicating similar degrees of myofiber membrane wounding by age. Despite this similarity, from genomic microarrays 318 unique transcripts were differentially expressed after RL in old vs. only 87 in young subjects. Follow-up pathways analysis and functional annotation revealed among old subjects upregulation of transcripts related to stress and cellular compromise, inflammation and immune responses, necrosis, and protein degradation and changes in expression (up- and downregulation) of transcripts related to skeletal and muscular development, cell growth and proliferation, protein synthesis, fibrosis and connective tissue function, myoblast-myotube fusion and cell-cell adhesion, and structural integrity. Overall the transcript-level changes indicative of undue inflammatory and stress responses in these older adults were not mirrored in young subjects. Follow-up immunoblotting revealed higher protein expression among old subjects for NF kappaB, heat shock protein (HSP)70, and IL-6 signaling [total and phosphorylated signal transducer and activator of transcription (STAT)3 at Tyr705]. Together, these novel findings suggest that young and old adults are equally susceptible to RL-mediated damage, yet the muscles of older adults are much more sensitive to this modest degree of damage-launching a robust transcriptome-level response that may begin to reveal key differences in the regenerative capacity of skeletal muscle with advancing age. PMID- 19903762 TI - Parent-of-origin effects on voluntary exercise levels and body composition in mice. AB - Despite the health-related benefits of exercise, many people do not engage in enough activity to realize the rewards, and little is known regarding the genetic or environmental components that account for this individual variation. We created and phenotyped a large G(4) advanced intercross line originating from reciprocal crosses between mice with genetic propensity for increased voluntary exercise (HR line) and the inbred strain C57BL/6J. G(4) females (compared to males) ran significantly more when provided access to a running wheel and were smaller with a greater percentage of body fat pre- and postwheel access. Change in body composition resulting from a 6-day exposure to wheels varied between the sexes with females generally regulating energy balance more precisely in the presence of exercise. We observed parent-of-origin effects on most voluntary wheel running and body composition traits, which accounted for 3-13% of the total phenotypic variance pooled across sexes. G(4) individuals descended from progenitor (F(0)) crosses of HRfemale symbol and C57BL/6Jmale symbol ran greater distances, spent more time running, ran at higher maximum speeds/day, and had lower percent body fat and higher percent lean mass than mice descended from reciprocal progenitor crosses (C57BL/6Jfemale symbol x HRmale symbol). For some traits, significant interactions between parent of origin and sex were observed. We discuss these results in the context of sex dependent activity and weight loss patterns, the contribution of parent-of-origin effects to predisposition for voluntary exercise, and the genetic (i.e., X-linked or mtDNA variations), epigenetic (i.e., genomic imprinting), and environmental (i.e., in utero environment or maternal care) phenomena potentially modulating these effects. PMID- 19903764 TI - Opposing effects of 5,7-DHT infusions into the orbitofrontal cortex and amygdala on flexible responding. AB - Central serotonin is implicated in a variety of emotional and behavioral control processes. Serotonin depletion can lead to exaggerated aversive processing and deficient response inhibition, effects that have been linked to serotonin's actions in the amygdala and orbitofrontal cortex (OFC), respectively. However, a direct comparison of serotonin manipulations within the OFC and amygdala in the same experimental context has not been undertaken. This study compared the effects of infusing the serotonin neurotoxin, 5,7-dihydroxytryptamine into the OFC and amygdala of marmosets performing an appetitive test of response inhibition. Marmosets had to learn to inhibit a prepotent response tendency to choose a box containing high-incentive food and instead choose a box containing low-incentive food, to obtain reward. OFC infusions caused long-lasting reductions in serotonin tissue levels, as revealed at postmortem, and exaggerated prepotent responses. In contrast, the significantly reduced prepotent responses following amygdala infusions occurred at a time when serotonin tissue levels had undergone considerable recovery, but there remained residual reductions in extracellular serotonin, in vivo. These opposing behavioral effects of serotonin manipulations in the same experimental context may be understood in terms of the top-down regulatory control of the amygdala by the OFC. PMID- 19903763 TI - Integrated approach for the comprehensive characterization of lipoproteins from human plasma using FPLC and nano-HPLC-tandem mass spectrometry. AB - The implication of the various lipoprotein classes in the development of atherosclerotic cardiovascular disease has served to focus a great deal of attention on these particles over the past half-century. Using knowledge gained by the sequencing of the human genome, recent research efforts have been directed toward the elucidation of the proteomes of several lipoprotein subclasses. One of the challenges of such proteomic experimentation is the ability to initially isolate plasma lipoproteins subsequent to their analysis by mass spectrometry. Although several methods for the isolation of plasma lipoproteins are available, the most commonly utilized techniques require large sample volumes and may cause destruction and dissociation of lipoprotein particle-associated proteins. Fast protein liquid chromatography (FPLC) is a nondenaturing technique that has been validated for the isolation of plasma lipoproteins from relatively small sample volumes. In this study, we present the use of FPLC in conjunction with nano-HPLC ESI-tandem mass spectrometry as a new integrated methodology suitable for the proteomic analysis of human lipoprotein fractions. Results from our analysis show that only 200 microl of human plasma suffices for the isolation of whole high density lipoprotein (HDL) and the identification of the majority of all known HDL associated proteins using mass spectrometry of the resulting fractions. PMID- 19903765 TI - Frequency facilitation at mossy fiber-CA3 synapses of freely behaving rats contributes to the induction of persistent LTD via an adenosine-A1 receptor regulated mechanism. AB - Frequency facilitation (FF), comprising a rapid and multiple-fold increase in the magnitude of evoked field potentials, is elicited by low-frequency stimulation (LFS) at mossy fiber-CA3 synapses. Here, we show that in freely behaving rats, FF reliably occurs in response to 1 and 2Hz but not in response to 0.25-, 0.3-, or 0.5-Hz LFS. Strikingly, prolonged (approximately 600 s) FF was tightly correlated to the induction of long-term depression (LTD) in freely moving animals. Although LFS at 2 Hz elicited unstable FF and unstable LTD, application of LFS at 1 Hz elicited pronounced FF, as well as robust LTD that persisted for over 24 h. This correlation of prolonged FF with LTD was absent at stimulation frequencies that did not induce FF. The adenosine-A1 receptor appears to participate in these effects: Application of adenosine-A1, but not adenosine-A3, receptor antagonists enhanced mossy fiber synaptic transmission and occluded FF. Furthermore, adenosine-A1 receptor antagonism resulted in more stable FF at 1 or 2 Hz and elicited more potent LTD. These data support the fact that FF contributes to the enablement of long-term information storage at mossy fiber-CA3 synapses and that the adenosine-A1 receptor may regulate the thresholds for this process. PMID- 19903766 TI - Nicotine enhances the antiapoptotic function of Mcl-1 through phosphorylation. AB - Lung cancer has a strong etiologic association with cigarette smoking. Nicotine, a major component in tobacco smoke, functions as a survival agonist that inhibits apoptosis following various stresses. However, the mechanism of action remains elusive. Mcl-1, a major antiapoptotic protein of the Bcl2 family, is extensively expressed in both small cell and non-small cell lung cancer cells, suggesting that Mcl-1 may be a therapeutic target of patients with lung cancer. Here, we found that nicotine induces Mcl-1 phosphorylation through activation of extracellular signal-regulated kinase 1/2 in association with increased chemoresistance of human lung cancer cells. Since nicotine stimulates Mcl-1 phosphorylation and survival in cells expressing wild-type but has no such effects in cells expressing T163A Mcl-1 mutant, this indicates that nicotine induces Mcl-1 phosphorylation exclusively at the T163 site and that phosphorylation of Mcl-1 at T163 is required for nicotine-induced survival. Mechanistically, nicotine-induced Mcl-1 phosphorylation significantly enhances the half-life of Mcl-1, which renders Mcl-1 a long-term survival activity. Specific depletion of Mcl-1 by RNA interference blocks nicotine-stimulated survival and enhances apoptotic cell death. Thus, nicotine-enhanced survival of lung cancer cells may occur through activation of Mcl-1 by phosphorylation at T163 site, which may contribute to development of human lung cancer and/or chemoresistance. PMID- 19903767 TI - Caveolin-1 promotes autoregulatory, Akt-mediated induction of cancer-promoting growth factors in prostate cancer cells. AB - Caveolin-1 (cav-1) and the cancer-promoting growth factors vascular endothelial growth factor (VEGF), transforming growth factor beta1 (TGF-beta1), and fibroblast growth factor 2 (FGF2) are often found to be upregulated in advanced prostate cancer and other malignancies. However, the relationship between cav-1 overexpression and growth factor upregulation remains unclear. This report presents, to our knowledge, the first evidence that in prostate cancer cells, a positive autoregulatory feedback loop is established in which VEGF, TGF-beta1, and FGF2 upregulate cav-1, and cav-1 expression, in turn, leads to increased levels of VEGF, TGF-beta1, and FGF2 mRNA and protein, resulting in enhanced invasive activities of prostate cancer cells, i.e., migration and motility. Our results further show that cav-1-enhanced mRNA stability is a major mechanism underlying the upregulation of these cancer-promoting growth factors, and that PI3-K-Akt signaling is required for forming this positive autoregulatory feedback loop. PMID- 19903769 TI - Critical role of endogenous heme oxygenase 1 as a tuner of the invasive potential of prostate cancer cells. AB - Prostate cancer (PCa) is the second leading cause of cancer-associated death in men. Inflammation has been recognized as a risk factor for this disease. Heme oxygenase 1 (HO-1), the inducible isoform of the rate-limiting enzyme in heme degradation, counteracts oxidative and inflammatory damage. Here, we investigated the regulated expression of HO-1 and its functional consequences in PCa. We studied the effect of genetic and pharmacologic disruption of HO-1 in the growth, invasion, and migration in androgen-sensitive (MDA PCa2b and LNCaP) and androgen insensitive (PC3) PCa cell lines. Our results show that HO-1 levels are markedly decreased in PC3 compared with MDA PCa2b and LNCaP. Hemin treatment increased HO 1 at both protein and mRNA levels in all cell lines and decreased cell proliferation and invasion. Furthermore, overexpression of HO-1 in PC3 resulted in markedly reduced cell proliferation and migration. Accordingly, small interfering RNA-mediated silencing of HO-1 expression in MDA PCa2b cells resulted in increased proliferation and invasion. Using reverse transcription-quantitative PCR-generated gene array, a set of inflammatory and angiogenic genes were upregulated or downregulated in response to HO-1 overexpression identifying matrix metalloprotease 9 (MMP9) as a novel downstream target of HO-1. MMP9 production and activity was downregulated by HO-1 overexpression. Furthermore, PC3 cells stably transfected with HO-1 (PC3HO-1) and controls were injected into nu/nu mice for analysis of in vivo tumor xenograft phenotype. Tumor growth and MMP9 expression was significantly reduced in PC3HO-1 tumors compared with control xenografts. Taken together, these results implicate HO-1 in PCa cell migration and proliferation suggesting its potential role as a therapeutic target in clinical settings. PMID- 19903768 TI - Upregulation of MMP-2 by HMGA1 promotes transformation in undifferentiated, large cell lung cancer. AB - Although lung cancer is the leading cause of cancer death worldwide, the precise molecular mechanisms that give rise to lung cancer are incompletely understood. Here, we show that HMGA1 is an important oncogene that drives transformation in undifferentiated, large-cell carcinoma. First, we show that the HMGA1 gene is overexpressed in lung cancer cell lines and primary human lung tumors. Forced overexpression of HMGA1 induces a transformed phenotype with anchorage independent cell growth in cultured lung cells derived from normal tissue. Conversely, inhibiting HMGA1 expression blocks anchorage-independent cell growth in the H1299 metastatic, undifferentiated, large-cell human lung carcinoma cells. We also show that the matrix metalloproteinase-2 (MMP-2) gene is a downstream target upregulated by HMGA1 in large-cell carcinoma cells. In chromatin immunoprecipitation experiments, HMGA1 binds directly to the MMP-2 promoter in vivo in large-cell lung cancer cells, but not in squamous cell carcinoma cells. In large-cell carcinoma cell lines, there is a significant, positive correlation between HMGA1 and MMP-2 mRNA. Moreover, interfering with MMP-2 expression blocks anchorage-independent cell growth in H1299 large-cell carcinoma cells, indicating that the HMGA1-MMP-2 pathway is required for this transformation phenotype in these cells. Blocking MMP-2 expression also inhibits migration and invasion in the H1299 large-cell carcinoma cells. Our findings suggest an important role for MMP-2 in transformation mediated by HMGA1 in large-cell, undifferentiated lung carcinoma and support the development of strategies to target this pathway in selected tumors. PMID- 19903770 TI - Activated platelets provide a functional microenvironment for the antiangiogenic fragment of histidine-rich glycoprotein. AB - The angiogenesis inhibitor histidine-rich glycoprotein (HRG) constitutes one of several examples of molecules regulating both angiogenesis and hemostasis. The antiangiogenic properties of HRG are mediated via its proteolytically released histidine- and proline-rich (His/Pro-rich) domain. Using a combination of immunohistochemistry and mass spectrometry, we here provide biochemical evidence for the presence of a proteolytic peptide, corresponding to the antiangiogenic domain of HRG, in vivo in human tissue. This finding supports a role for HRG as an endogenous regulator of angiogenesis. Interestingly, the His/Pro-rich peptide bound to the vessel wall in tissue from cancer patients but not to the vasculature in tissue from healthy persons. Moreover, the His/Pro-rich peptide was found in close association with platelets. Relesate from in vitro-activated platelets promoted binding of the His/Pro-rich domain of HRG to endothelial cells, an effect mediated by Zn(2+). Previous studies have shown that zinc dependent binding of the His/Pro-rich domain of HRG to heparan sulfate on endothelial cells is required for inhibition of angiogenesis. We describe a novel mechanism to increase the local concentration and activity of an angiogenesis inhibitor, which may reflect a host response to counteract angiogenesis during pathologic conditions. Our finding that tumor angiogenesis is elevated in HRG deficient mice supports this conclusion. PMID- 19903771 TI - The association and nuclear translocation of the PIAS3-STAT3 complex is ligand and time dependent. AB - The epidermal growth factor (EGF) receptor activation of downstream signal transducers and activators of transcription 3 (STAT3) plays a crucial role in the pathogenesis of lung cancer. STAT3 transcriptional activity can be negatively regulated by protein inhibitor of activated STAT3 (PIAS3). We investigated the time-dependent PIAS3 shuffling and binding to STAT3 in an EGF-dependent model in lung cancer by using confocal microscopy, immunoprecipitation, luciferase reporter assay, and protein analysis of segregated cellular components. We also explored the role of phosphorylation at Tyr705 of STAT3 in the formation and intracellular shuffling of the PIAS3-STAT3 complex. In a growth factor-free state, PIAS3 was localized to the cytoplasm and unbound to STAT3 in both H520 and A549 cells. On exposure to EGF, we observed STAT3 phosphorylation and rapid formation of the PIAS3-STAT3 complex. Within 5 minutes, there was a progressive translocation of the complex to the nucleus, and by 10 minutes, PIAS3 was uniquely localized to the nuclear compartment. After 30 minutes, PIAS3 returned to the cytoplasm. Using site-directed mutagenesis, we substituted Tyr705 of STAT3 with a phenylalanine. Despite EGF stimulation, we observed a significant decrease in PIAS3-STAT3 binding and a significant reduction in nuclear translocation of PIAS3. Furthermore, there was a significant reduction in the capacity of PIAS3 to reduce STAT3-mediated gene transcription. In wild-type STAT3 cells, increasing concentrations of PIAS3 resulted in a proportional decrease in STAT3 phosphorylation. These data suggest an important role for the negative regulatory effect of PIAS3 on STAT3 in EGF-driven tumors. PMID- 19903772 TI - p34SEI-1 inhibits doxorubicin-induced senescence through a pathway mediated by protein kinase C-delta and c-Jun-NH2-kinase 1 activation in human breast cancer MCF7 cells. AB - In this study, we describe a novel function of the p34(SEI-1) protein, which is both an oncogenic protein and a positive regulator of the cell cycle. The p34(SEI 1) protein was found to inhibit doxorubicin-induced senescence. We investigated the molecular mechanisms of the inhibitory effect of p34(SEI-1) on senescence. First, we found that the activation of protein kinase C-delta (PKC-delta), which is cleaved into a 38 kDa active form from a 78 kDa pro-form, induced after doxorubicin treatment, was inhibited by p34(SEI-1). Furthermore, p34(SEI-1) induced the ubiquitination of PKC-delta. Yet, there is no interaction between p34(SEI-1) and PKC-delta. We also found that the phosphorylation of c-Jun-NH(2) kinase 1 (JNK1) induced after doxorubicin treatment was suppressed by p34(SEI-1), but not in JNK2. Consistently, pharmacologic or genetic inactivation of either PKC-delta or JNK1 was found to inhibit doxorubicin-induced senescence. In addition, the genetic inactivation of PKC-delta by PKC-delta small interfering RNA resulted in an inhibition of JNK1 activation, but PKC-delta expression was not inactivated by JNK1 small interfering RNA, implying that the activation of JNK1 could be dependently induced by PKC-delta. Therefore, p34(SEI-1) inhibits senescence by inducing PKC-delta ubiquitination and preventing PKC-delta dependent phosphorylation of JNK1. PMID- 19903773 TI - Occurrence of bacteria and viruses on elementary classroom surfaces and the potential role of classroom hygiene in the spread of infectious diseases. AB - The presence of microorganisms on common classroom contact surfaces (fomites) was determined to identify the areas most likely to become contaminated. Six elementary classrooms were divided into control and intervention groups (cleaned daily with a quaternary ammonium wipe) and tested for heterotrophic bacteria. Three classrooms were also tested for norovirus and influenza A virus. Frequently used fomites were the most contaminated; water fountain toggles, pencil sharpeners, keyboards, and faucet handles were the most bacterially contaminated; desktops, faucet handles, and paper towel dispensers were the most contaminated with viruses. Influenza A virus was detected on up to 50% and norovirus on up to 22% of surfaces throughout the day. Children in the control classrooms were 2.32 times more likely to report absenteeism due to illness than children in the intervention classrooms and were absent longer (on average). Improved classroom hygiene may reduce the incidence of infection and thus student absenteeism. PMID- 19903774 TI - Physical activity, alcohol and tobacco use and associated cardiovascular morbidity and mortality in the Second Australian National Blood Pressure study cohort. PMID- 19903775 TI - Health status and risk for depression among the elderly: a meta-analysis of published literature. AB - OBJECTIVE: the goal of this study was to determine the relationship between health status, including self-rated health status and chronic disease, and risk for depression among the elderly. METHOD: MEDLINE, EMBASE and The Cochrane Library Database were used to identify potential studies. The studies were classified into cross-sectional and longitudinal subsets. For each study, the numbers of the total participants, cases (for cross-sectional study) or incident cases (for longitudinal study) of depression in each health status group were extracted and entered into Review Manager 4.2. The quantitative meta-analysis of cross-sectional studies and that of longitudinal studies were performed, respectively. For prevalence and incidence rates of depression, odds risk and relative risk (RR) were calculated, respectively. RESULTS: the quantitative meta analysis showed that, compared with the elderly without chronic disease, those with chronic disease had higher risk for depression (RR: 1.53, 95% confidence intervals (CI): 1.20-1.97). Compared with the elderly with good self-rated health, those with poor self-rated health had higher risk for depression (RR: 2.40, 95% CI: 1.94-2.97). CONCLUSIONS: despite the methodological limitations of this meta-analysis, both poor self-rated health status and the presence of chronic disease are risk factors for depression among the elderly. In the elderly, poor self-reported health status appears to be more strongly associated with depression than the presence of chronic disease. PMID- 19903776 TI - Contribution of degradation products to the anticancer activity of curcumin. PMID- 19903777 TI - Effect of granulocyte/macrophage colony-stimulating factor on vaccination with an allogeneic whole-cell melanoma vaccine. AB - PURPOSE: The availability of a variety of immune response modifiers creates an opportunity for improved efficacy of immunotherapy, but it also leads to uncertainty in how to combine agents and how to assess those combinations. We sought to assess the effect of the addition of granulocyte/macrophage colony stimulating factor (GM-CSF) to vaccination with a melanoma vaccine. EXPERIMENTAL DESIGN: Ninety-seven patients with resected melanoma (stage II-IV) were enrolled, stratified by stage, and randomized to receive a cellular melanoma vaccine with or without GM-CSF. The primary endpoint was delayed-type hypersensitivity (DTH) response to melanoma cells. Antibody responses, peripheral leukocyte counts, and survival were also examined. RESULTS: The GM-CSF arm showed enhanced antibody responses with an increase in IgM titer against the TA90 antigen and increased TA90 immune complexes. This arm also had diminished antimelanoma cell delayed type hypersensitivity response. Peripheral blood leukocyte profiles showed increases in eosinophils and basophils with decreased monocytes in the GM-CSF arm. These immune changes were accompanied by an increase in early melanoma deaths and a trend toward worse survival with GM-CSF. CONCLUSION: These data suggest that GM-CSF is not helpful as an immune adjuvant in this dose and schedule and raise concern that it may be harmful. Based on the discordant findings of an immune endpoint and clinical outcome, the use of such surrogate endpoints in selecting treatments for further evaluation must be done with a great deal of caution. PMID- 19903779 TI - Immunostimulation versus immunosuppression after multiple vaccinations: the woes of therapeutic vaccine development. AB - Three articles in this issue of Clinical Cancer Research show how multiple vaccinations can lead to immunosuppression. Moreover, two studies in patients show that granulocyte macrophage colony-stimulating factor (GM-CSF) as an adjuvant immunostimulant to different kind of vaccines can lead to adverse outcome in terms of relapse-free and overall survival. Modulation of regulatory T cell activity may be required to overcome this outcome and may be crucial for the successful development of therapeutic vaccines. PMID- 19903778 TI - Phase I trial of a combination of the multikinase inhibitor sorafenib and the farnesyltransferase inhibitor tipifarnib in advanced malignancies. AB - PURPOSE: We evaluated the safety, maximum tolerated dose, pharmacokinetics, and biological effects of the combination of the Raf-1, RET, KIT, platelet-derived growth factor receptor, and vascular endothelial growth factor receptor 2 kinase inhibitor sorafenib and the farnesyltransferase inhibitor tipifarnib. EXPERIMENTAL DESIGN: A standard 3 + 3 phase I dose-escalation design was used with a 28-day cycle (sorafenib daily and tipifarnib for 21 days, by mouth). RESULTS: Fifty patients were treated; 43 reached restaging evaluation after cycle 2. The most common side effects were grade 1 to 2 rash, hyperglycemia, and diarrhea. Dose-limiting toxicity was rash, and the recommended phase II dose is sorafenib 400 mg p.o. qam/200 mg p.o. qpm and tipifarnib p.o. 100 mg bd. Despite the low doses of tipifarnib, one quarter of patients had > or =50% reduction in farnesyltransferase levels. Interestingly, six of eight patients with medullary thyroid cancer had durable stable disease (n = 3) or partial remissions (n = 3), lasting 12 to 26+ months. Five of the six responders had available tissue, and RET gene mutations were identified in them. Prolonged (> or =6 months) stable disease was also seen in nine patients as follows: papillary thyroid cancer (n = 4; 18+ to 27+ months), adrenocortical cancer (n = 2; 7 and 11 months), and one each of melanoma (platelet-derived growth factor receptor mutation positive; 14 months), renal (6 months), and pancreatic cancer (6 months). CONCLUSIONS: Our study shows that the combination of tipifarnib and sorafenib is well tolerated. Activity was seen, especially in patients with medullary thyroid cancer, a tumor characterized by RET mutations. PMID- 19903780 TI - Effect of granulocyte/macrophage colony-stimulating factor on circulating CD8+ and CD4+ T-cell responses to a multipeptide melanoma vaccine: outcome of a multicenter randomized trial. AB - PURPOSE: Granulocyte/macrophage colony-stimulating factor (GM-CSF) administered locally together with vaccines can augment T-cell responses in animal models. Human experience has been limited to small and uncontrolled trials. Thus, a multicenter randomized phase II trial was done to determine whether local administration of GM-CSF augments immunogenicity of a multipeptide vaccine. It also assessed immunogenicity of administration in one versus two vaccine sites. EXPERIMENTAL DESIGN: One hundred twenty-one eligible patients with resected stage IIB to IV melanoma were vaccinated with 12 MHC class I-restricted melanoma peptides to stimulate CD8+ T cells plus a HLA-DR-restricted tetanus helper peptide to stimulate CD4+ T cells, emulsified in incomplete Freund's adjuvant, with or without 110 microg GM-CSF. Among 119 evaluable patients, T-cell responses were assessed by IFN-gamma ELIspot assay and tetramer analysis. Clinical outcomes were recorded. RESULTS: CD8+ T-cell response rates to the 12 MHC class I restricted melanoma peptides (by day 50) with or without GM-CSF were 34% and 73%, respectively (P < 0.001), by direct ELIspot assay. Tetramer analyses corroborated the functional data. CD4+ T-cell responses to tetanus helper peptide were higher without GM-CSF (95% versus 77%; P = 0.005). There was no significant difference by number of vaccine sites. Three-year overall and disease-free survival estimates (95% confidence interval) were 76% (67-83%) and 52% (43-61%), respectively, with too few events to assess differences by study group. CONCLUSIONS: High immune response rates for this multipeptide vaccine were achieved, but CD8+ and CD4+ T-cell responses were lower when administered with GM CSF. These data challenge the value of local GM-CSF as a vaccine adjuvant in humans. PMID- 19903782 TI - Survivin as a prognostic marker for urothelial carcinoma of the bladder: a multicenter external validation study. AB - PURPOSE: The aim of the current study was to externally validate the value of survivin as a prognostic marker for bladder cancer in a large multi-institutional cohort of patients treated with radical cystectomy. METHODS: The study comprised 726 patients treated with radical cystectomy and bilateral pelvic lymphadenectomy. Survivin staining and scoring were done with automated systems coupled with advanced color detection software. Specimens showing at least 10% reactivity were considered altered. Predictive accuracy was quantified using the concordance index and 200-bootstrap resamples were used to reduce overfit bias. RESULTS: Survivin was an independent predictor of disease recurrence and cancer specific survival in multivariable analyses that controlled for the effects of standard clinicopathologic features (hazard ratios, approximately 1.6; P values < or = 0.002). In all patients (n = 726), addition of survivin to a model including standard clinicopathologic variables did not improve its predictive accuracy (P = 0.67 for disease recurrence and P = 0.27 for cancer-specific survival). In the subgroup of patients with pT(1-3)N(0)M(0) disease (n = 398), addition of survivin improved the accuracy of standard clinicopathologic features for prediction of disease recurrence and cancer-specific survival (1.3%, P < 0.001 and 1.2%, P < 0.001, respectively). CONCLUSIONS: Survivin expression improves our accuracy for prediction of cancer recurrence and survival in pT(1-3)N(0)M(0) patients by a small but statistically significant margin. Our findings support the need for further evaluation of survivin and its signaling pathways as well as survivin targeted therapies in bladder cancer. PMID- 19903783 TI - Epidermal growth factor receptor is required for colonic tumor promotion by dietary fat in the azoxymethane/dextran sulfate sodium model: roles of transforming growth factor-{alpha} and PTGS2. AB - PURPOSE: Colon cancer is a major cause of cancer deaths. Dietary factors contribute substantially to the risk of this malignancy. Western-style diets promote development of azoxymethane-induced colon cancer. Although we showed that epidermal growth factor receptors (EGFR) controlled azoxymethane tumorigenesis in standard fat conditions, the role of EGFR in tumor promotion by high dietary fat has not been examined. EXPERIMENTAL DESIGN: A/J x C57BL6/J mice with wild-type Egfr (Egfr(wt)) or loss-of-function waved-2 Egfr (Egfr(wa2)) received azoxymethane followed by standard (5% fat) or western-style (20% fat) diet. As F(1) mice were resistant to azoxymethane, we treated mice with azoxymethane followed by one cycle of inflammation-inducing dextran sulfate sodium to induce tumorigenesis. Mice were sacrificed 12 weeks after dextran sulfate sodium. Tumors were graded for histology and assessed for EGFR ligands and proto-oncogenes by immunostaining, Western blotting, and real-time PCR. RESULTS: Egfr(wt) mice gained significantly more weight and had exaggerated insulin resistance compared with Egfr(wa2) mice on high-fat diet. Dietary fat promoted tumor incidence (71.2% versus 36.7%; P < 0.05) and cancer incidence (43.9% versus 16.7%; P < 0.05) only in Egfr(wt) mice. The lipid-rich diet also significantly increased tumor and cancer multiplicity only in Egfr(wt) mice. In tumors, dietary fat and Egfr(wt) upregulated transforming growth factor-alpha, amphiregulin, CTNNB1, MYC, and CCND1, whereas PTGS2 was only increased in Egfr(wt) mice and further upregulated by dietary fat. Notably, dietary fat increased transforming growth factor-alpha in normal colon. CONCLUSIONS: EGFR is required for dietary fat-induced weight gain and tumor promotion. EGFR-dependent increases in receptor ligands and PTGS2 likely drive diet-related tumor promotion. PMID- 19903784 TI - Partial CD4 depletion reduces regulatory T cells induced by multiple vaccinations and restores therapeutic efficacy. AB - PURPOSE: A single vaccination of intact or reconstituted-lymphopenic mice (RLM) with a granulocyte macrophage colony-stimulating factor-secreting B16BL6-D5 melanoma cell line induces protective antitumor immunity and T cells that mediate the regression of established melanoma in adoptive immunotherapy studies. We wanted to study if multiple vaccinations during immune reconstitution of the lymphopenic host would maintain a potent antitumor immune response. EXPERIMENTAL DESIGN: RLM were vaccinated multiple times over a 40-day period. Spleens were isolated from these mice, activated in vitro, and adoptively transferred into mice bearing 3-day experimental pulmonary metastases. RESULTS: Multiple vaccinations, rather than boosting the immune response, significantly reduced therapeutic efficacy of adoptive immunotherapy and were associated with an increased frequency and absolute number of CD3+CD4+Foxp3+ T regulatory (T(reg)) cells. Anti-CD4 administration reduced the absolute number of T(reg) cells 9 fold. Effector T-cells generated from anti-CD4-treated mice were significantly (P < 0.0001) more therapeutic in adoptive transfer studies than T cells from multiply vaccinated animals with a full complement of CD4+ cells. CONCLUSION: These results suggest that CD4+ T(reg) cells limit the efficacy of multiple vaccinations and that timed partial depletion of CD4+ T cells may reduce suppression and "tip-the-balance" in favor of therapeutic antitumor immunity. The recent failure of large phase III cancer vaccine clinical trials, wherein patients received multiple vaccines, underscores the potential clinical relevance of these findings. PMID- 19903785 TI - A phase 1 dose escalation study of the safety and pharmacokinetics of the novel proteasome inhibitor carfilzomib (PR-171) in patients with hematologic malignancies. AB - PURPOSE: Carfilzomib (formerly PR-171) is a novel proteasome inhibitor of the epoxyketone class that is selective and structurally distinct from bortezomib. Proteasome inhibition by carfilzomib is mechanistically irreversible. Consequently, proteasome inhibition is more sustained with carfilzomib than with bortezomib. EXPERIMENTAL DESIGN: In a phase 1 trial evaluating the safety and efficacy of carfilzomib in relapsed or refractory hematologic malignancies, eight dose groups of three to six patients received 5 consecutive days of carfilzomib i.v. push at doses of 1.2, 2.4, 4, 6, 8.4, 11, 15, and 20 mg/m2 within 14-day cycles. RESULTS: Twenty-nine patients enrolled that were relapsed or refractory after at least two prior therapies. Nonhematologic toxicities included fatigue, nausea, and diarrhea in more than one third of patients-mostly grade 1 or 2 in severity. At 20 mg/m2, grade 3 febrile neutropenia and grade 4 thrombocytopenia were reported, establishing 15 mg/m2 as the maximum tolerated dose. No grade 3 or 4 peripheral neuropathies were reported. Antitumor activity was observed at doses > or =11 mg/m2: one unconfirmed complete response (mantle cell), one partial response (multiple myeloma), and two minimal responses (multiple myeloma and Waldenstrom's macroglobulinemia). CONCLUSION: This is the first clinical use of carfilzomib that shows tolerability and clinical activity in multiple hematologic malignancies using consecutive-day dosing. PMID- 19903786 TI - PIK3CA mutations predict local recurrences in rectal cancer patients. AB - PURPOSE: Identifying rectal cancer patients at risk for local recurrence would allow for refinement in the selection of patients who would benefit from preoperative radiotherapy. PIK3CA, KRAS, and BRAF mutations are commonly found in colon cancers, but their prevalence has not been clearly assessed in rectal cancer. In this study, we aim to determine the mutation frequencies of PIK3CA, KRAS, and BRAF and to investigate whether a mutation may be used as a prognostic parameter in rectal cancer patients. EXPERIMENTAL DESIGN: We evaluated DNA mutations in PIK3CA, KRAS, and BRAF in 240 stage I to III rectal tumors obtained from nonirradiated patients from the Dutch Total Mesorectal Excision trial. RESULTS: PIK3CA, KRAS, and BRAF mutations were identified in 19 (7.9%), 81 (33.9%), and 5 (2.1%) rectal cancers. Patients with PIK3CA mutations developed more local recurrences (5-year risks, 27.8% versus 9.4%; P = 0.006) and tended to develop these recurrences more rapidly after surgery (median local recurrence free interval since surgery: 7.9 versus 19.6 months; P = 0.07) than patients without PIK3CA mutations. In multivariate analysis, PIK3CA mutations remained as an independent predictor for the development of local recurrences (hazard ratio, 3.4; 95% confidence interval, 1.2-9.2; P = 0.017), next to tumor-node-metastasis stage. CONCLUSION: PIK3CA mutations can be used as a biomarker in identifying rectal cancer patients with an increased risk for local recurrences. Currently, our findings suggest that prospective evaluation of PIK3CA mutation status could reduce overtreatment by preoperative radiotherapy for the low-risk patients who might otherwise only experience the side effects. PMID- 19903787 TI - Electrocardiographic characterization of the QTc interval in patients with advanced solid tumors: pharmacokinetic- pharmacodynamic evaluation of sunitinib. AB - PURPOSE: To evaluate the effects of sunitinib, a multitargeted tyrosine kinase inhibitor, on the QT interval in patients with cancer. EXPERIMENTAL DESIGN: Patients received sunitinib loading doses (150-200 mg) on days 3 and 9 and maintenance doses (50 mg/d) on days 4 to 8. Moxifloxacin (day 1), placebo (day 2), and granisetron [with placebo (day 2) or sunitinib (days 3 and 9)] were also administered. Treatment effects were evaluated by time-matched, serial electrocardiograms, and manually overread. RESULTS: Twenty-four of 48 patients were QT/PK evaluable. Moxifloxacin produced a time-matched, maximum mean placebo adjusted corrected QT interval (QT(c)F) of 5.6 ms [90% confidence interval (CI), 1.9-9.3]. Sunitinib QT(c)F changes correlated with exposure, but not T(max). Maximum mean time-matched, placebo-adjusted QT(c)F was 9.6 ms (90% CI, 4.1-15.1) at steady state/therapeutic concentrations (day 3) and 15.4 ms (90% CI, 8.4-22.4) at supratherapeutic concentrations (day 9). No patient had a QT(c)F >500 ms. Concomitant granisetron produced no significant QT(c)F prolongation. Sunitinib related adverse events were as previously described. CONCLUSIONS: Sunitinib has a dose-dependent effect on QT interval. The increased risk of ventricular arrhythmias must be weighed against the therapeutic benefit sunitinib provides to patients with advanced cancer. PMID- 19903789 TI - Deregulated GSK3{beta} sustains gastrointestinal cancer cells survival by modulating human telomerase reverse transcriptase and telomerase. AB - PURPOSE: Glycogen synthase kinase-3beta (GSK3beta) regulates multiple cell signaling pathways and has been implicated in glucose intolerance, neurodegenerative disorders, and inflammation. We investigated the expression, activity, and putative pathologic role of GSK3beta in gastrointestinal, pancreatic, and liver cancers. EXPERIMENTAL DESIGN: Colon, stomach, pancreatic, and liver cancer cell lines; nonneoplastic HEK293 cells; and matched pairs of normal and tumor tissues of stomach and colon cancer patients were examined for GSK3beta expression and its phosphorylation at serine 9 (inactive form) and tyrosine 216 (active form) by Western immunoblotting and for GSK3beta activity by in vitro kinase assay. The effects of small-molecule GSK3beta inhibitors and of RNA interference on cell survival, proliferation, and apoptosis were examined in vitro and on human colon cancer cell xenografts in athymic mice. The effects of GSK3beta inhibition on human telomerase reverse transcriptase (hTERT) expression and telomerase activity were compared between colon cancer and HEK293 cells. RESULTS: Cancer cell lines and most cancer tissues showed increased GSK3beta expression and increased tyrosine 216 phosphorylation and activity but decreased serine 9 phosphorylation compared with HEK293 cells and nonneoplastic tissues. Inhibition of GSK3beta resulted in attenuated cell survival and proliferation and increased apoptosis in most cancer cell lines and in HT-29 xenografts in rodents but not in HEK293 cells. GSK3beta inhibition in colon cancer cells was associated with decreased hTERT expression and telomerase activity. CONCLUSION: The results indicate that deregulated GSK3beta sustains gastrointestinal cancer cells survival through modulation of hTERT and telomerase. PMID- 19903788 TI - Subsets of very low risk Wilms tumor show distinctive gene expression, histologic, and clinical features. AB - PURPOSE: Recent studies suggest that children <24 months with stage I favorable histology Wilms tumors <550 g [very low risk Wilms tumors (VLRWT)] have an excellent prognosis when treated with nephrectomy only, without adjuvant chemotherapy. The identification of risk categories within VLRWT may enable refinement of their definition and optimization of their therapy. EXPERIMENTAL DESIGN: To define biologically distinct subsets, global gene expression analysis was done on 39 VLRWT that passed all quality-control parameters and the clusters identified were validated in an independent set of 11 VLRWT. Validation of select differentially expressed genes was done with immunohistochemistry on a tissue microarray from 20 of 39 tumors. Loss of heterozygosity (LOH) for 11p15, 1p, and 16q was analyzed in 52 tumors using PCR. RESULTS: Two distinctive clusters were identified. One cluster included 9 tumors with epithelial differentiated tubular histology, paucity of nephrogenic rests, lack of LOH for 1p, 16q, and 11p, absence of relapse, and a unique gene expression profile consistent with arrest following mesenchymal-to-epithelial transition. The second cluster included 13 tumors with mixed histology, intralobar nephrogenic rests, and decreased expression of WT1. Three of 6 relapses occurred in this cluster. Of 43 informative tumors, 11p LOH was present in 5 of 5 relapses and 11 of 38 nonrelapses. CONCLUSIONS: Two subsets comprising a total of 56% of VLRWT are identified that have pathogenetic and molecular differences and apparent differences in risk for relapse. If these predictors can be prospectively validated, this would enable the refinement of clinical stratification and less arbitrary definition of VLRWT. PMID- 19903790 TI - Getting knit-PI3Ky: PIK3CA mutation status to direct multimodality therapy? AB - There is high morbidity associated with local recurrence of rectal cancer. However, the adjuvant therapies given to prevent such recurrences also have significant side effects and associated risks. The ability to select patients with the highest risk of recurrence and greatest therapeutic response will improve rectal cancer care. PMID- 19903791 TI - A novel toll-like receptor 9 agonist cooperates with trastuzumab in trastuzumab resistant breast tumors through multiple mechanisms of action. AB - PURPOSE: Resistance to anti-HER2 monoclonal antibody trastuzumab is a relevant issue in breast cancer patients. Among the mechanisms implicated in trastuzumab resistance, increasing evidence supports a role of tumor microenvironment. We previously found that a novel toll-like receptor 9 agonist, referred to as immune modulatory oligonucleotide (IMO) and currently under clinical investigation, acts through epidermal growth factor receptor (EGFR) and shows direct antiangiogenic effects by cooperating with anti-EGFR or anti-VEGF drugs, thus interfering with cancer cells and microenvironment. EXPERIMENTAL DESIGN: In this study, we used KPL-4 and JIMT-1 trastuzumab-resistant breast cancer cells to evaluate the combination IMO plus trastuzumab as a therapeutic option for trastuzumab resistant breast cancers. RESULTS: IMO inhibits KPL-4 and JIMT-1 xenografts growth and potentiates trastuzumab antitumor effect, with complete suppression of tumor growth, potent enhancement of trastuzumab-mediated antibody-dependent cell mediated cytotoxicity, and strong inhibition of EGFR/HER2-related signaling. In KPL-4 xenografts, IMO alone interferes with HER signal transduction, whereas trastuzumab is ineffective. IMO induces an HER-dependent signal inhibition also in vitro by modulating a functional interaction between toll-like receptor 9 and HER receptors occurring at membrane level. Finally, IMO plus trastuzumab produces a cooperative antiangiogenic effect related to suppression of endothelial HER related signaling. CONCLUSIONS: We showed a cooperative effect of IMO plus trastuzumab in trastuzumab-resistant breast cancers due to IMO direct antitumor and antiangiogenic activity and antibody-dependent cell-mediated cytotoxicity enhancement. Moreover, we provided first evidence of a toll-like receptor 9/HER interaction at membrane level as novel mechanism of action. Altogether, we propose IMO plus trastuzumab as an effective strategy in trastuzumab-resistant breast cancers. PMID- 19903792 TI - HIF-1alpha and HIF-2alpha are differentially regulated in vivo in neuroblastoma: high HIF-1alpha correlates negatively to advanced clinical stage and tumor vascularization. AB - PURPOSE: Hypoxia is considered to be a major driving force behind tumor angiogenesis. The stabilization and activation at hypoxia of the hypoxia inducible factors HIF-1alpha and HIF-2alpha and the concomitant induction of expression of vascular endothelial growth factor (VEGF) and other proangiogenic factors provide a molecular frame for hypoxia-driven tumor angiogenesis. This study has investigated how HIF and VEGF protein levels relate to each other with regard to vascularization, tumor stage, and overall survival in neuroblastoma. EXPERIMENTAL DESIGN: Tissue cores taken from tumor specimens representing 93 children with neuroblastoma were arranged on a microarray and stained for HIF 1alpha, HIF-2alpha, VEGF, and CD31 proteins. Both fraction of positive cells and staining intensity were evaluated and protein levels were correlated with each other and with clinical variables. RESULTS: Although high levels of both HIF 1alpha (P < 0.001) and HIF-2alpha (P < 0.001) correlated positively to VEGF expression, they did not fully correlate with each other. Moreover, HIF-1alpha (P = 0.002) and VEGF (P < 0.001), but not HIF-2alpha, correlated negatively to vascularization as determined by CD31 staining abundance. VEGF expression or degree of vascularization did not correlate with tumor stage or overall survival. High HIF-1alpha levels correlated with low tumor stage (P < 0.001) and were associated with a favorable patient prognosis (P = 0.08). CONCLUSIONS: The discordant results on expression of HIF-1alpha and HIF-2alpha suggest that these two proteins are differentially regulated in vivo, thus reflecting distinctive protein expression/stabilization mechanisms. The association between HIF-1alpha and favorable outcome stresses the importance of discriminating HIF-2alpha from HIF-1alpha expression and has implications for using HIFs as treatment targets. PMID- 19903794 TI - T-cell responses to cyclin B1 are not restricted to p53-overexpressing tumors. PMID- 19903795 TI - Isolated Addison's disease is unlikely to be caused by mutations in MC2R, MRAP or STAR, three genes responsible for familial glucocorticoid deficiency. AB - BACKGROUND: Familial glucocorticoid deficiency (FGD) is a rare autosomal recessive disease caused by ACTH resistance and leads to isolated glucocorticoid deficiency. Although FGD patients typically have normal mineralocorticoid secretion, subtle alterations in the renin-angiotensin-aldosterone axis have been reported in a subset of patients at presentation. Anecdotally, some patients with FGD have been initially diagnosed as having Addison's disease (AD), with implications for treatment and genetic counselling. Currently, mutations in three genes: the ACTH receptor (MC2R); the melanocortin 2 receptor accessory protein (MRAP); and the steroidogenic acute regulatory protein (STAR) are known to give rise to FGD types 1-3. We investigated a cohort of autoantibody-negative AD patients for mutations in these genes. METHODS: Forty patients with known AD without evidence of autoimmune disease were screened for mutations in MC2R, MRAP and STAR. In addition, patients were genotyped for the MC2R promoter polymorphism previously associated with reduced responsiveness to ACTH. RESULTS: No mutations in MC2R, MRAP or STAR were identified in any patient. The frequencies of the MC2R promoter polymorphism were similar to those reported in healthy controls. CONCLUSIONS: FGD does not appear to be underdiagnosed in the AD population. However, in approximately 50% of patients with FGD, no genetic cause has yet been identified and it is possible that the other, as yet unidentified, genes giving rise to FGD may be implicated in AD. PMID- 19903796 TI - Bevacizumab plus capecitabine as a salvage therapy in advanced adrenocortical carcinoma. AB - OBJECTIVE: No standard therapy for advanced adrenocortical carcinoma (ACC) is established by any randomized trial but a consensus conference 2003 recommended mitotane as monotherapy or combined with etoposide, doxorubicin and cisplatin or with streptozotocin as first-line systemic therapy. However, there is no evidence for any therapy beneficial in patients failing these therapies. Therefore, we evaluated the effects of the anti-VEGF antibody bevacizumab plus capecitabine as salvage therapy in ACC. METHODS: Patients registered with the German ACC Registry with refractory ACC progressing after cytotoxic therapies were offered treatment with bevacizumab (5 mg/kg body weight i.v. every 21 days) and oral capecitabine (950 mg/m(2) twice daily for 14 days followed by 7 days of rest) in 2006-2008. Evaluation of tumour response was performed by imaging according to response evaluation criteria in solid tumours every 12 weeks. RESULTS: Ten patients were treated with bevacizumab plus capecitabine. None of them experienced any objective response or stable disease. Two patients had to stop therapy after few weeks due to hand-foot syndrome, and three patients died on progressive disease within 12 weeks. Other adverse events were mild (grade I-II). Median survival after treatment initiation was 124 days. CONCLUSIONS: Bevacizumab plus capecitabine has no activity in patients with very advanced ACC. Hence, this regimen cannot be recommended as a salvage therapy. PMID- 19903797 TI - Effects of long-term L-thyroxine treatment on endothelial function and arterial distensibility in young adults with congenital hypothyroidism. AB - OBJECTIVE: Patients with congenital hypothyroidism (CH) display subclinical abnormalities of the cardiovascular system that are related to unphysiological fluctuations of TSH levels and occur despite careful replacement therapy. DESIGN: The aim of the present case-control study was to evaluate the effects of long term levothyroxine (l-T(4)) replacement therapy on the vascular district in CH patients by assessing endothelial function with flow-mediated dilation (FMD) and brachial artery distensibility with the measurement of the coefficient of distensibility (DC). METHODS: Thirty-two young adults with CH aged 18.9+/-0.2 years and 32 age- and sex-matched controls underwent brachial Doppler ultrasound examination to measure FMD and DC at the time of the study. Hypothyroidism was diagnosed by neonatal screening, and l-T(4) treatment was initiated within the first month of life. RESULTS: Compared to healthy controls, CH patients had significantly reduced brachial artery reactivity with lower FMD values (8.9+/-5.7 vs 14.1+/-5.1% P=0.003) and decreased vascular distensibility (24.6+/-1.6 vs 27.3+/-3 kPa(-1)x10(-3), P<0.0002). Linear regression analysis revealed that both total and pubertal mean TSH and number of episodes of undertreatment were independent determinants of FMD and DC. Pubertal mean TSH was the best predictor of both FMD and DC (r=0.81 and r=0.87 respectively, P<0.001). CONCLUSIONS: Young adults with CH treated with long-term l-T(4) replacement therapy may have significant impairment of both FMD and DC. Our data suggest that high TSH levels, inadequately corrected by l-T(4) replacement therapy in CH patients especially during puberty, can exert significant effects on the elastic and functional vessel properties. PMID- 19903798 TI - Hypothalamic-pituitary-adrenal axis response to the severity of illness in non critically ill patients: does relative corticosteroid insufficiency exist? AB - OBJECTIVE: Relative corticosteroid insufficiency may be common in critically ill patients and is associated with a poor outcome; however, the response of the hypothalamic-pituitary-adrenal (HPA) axis in nursed patients is not known. Our aim was to evaluate the response of HPA axis to the severity of illness in non critically ill nursed (NCIN) patients and the clinical outcome. SUBJECTS AND METHODS: Fifty-six nursed patients who were divided into four groups (stroke, mild disease, sepsis and severe sepsis) as well as a control group (n=15) were studied. At admission (day 1), cortisol and ACTH were measured and a low-dose (1 microg) corticotrophin test was performed, followed 2 h later by a standard-dose (250 microg) corticotrophin test. Diurnal variation of cortisol was obtained on day 2. A second identical set of low-dose and standard-dose corticotrophin tests were performed on day 5 or 6 (recovery phase). Results In patients with stroke and severe sepsis, cortisol had the highest values and its diurnal variation was abolished. Dissociation of ACTH and cortisol was found in all patients. The Delta(max) of cortisol after the 1-microg corticotrophin test did not differ among the groups, while after the 250-microg corticotrophin test, it was borderline higher in controls. The ratio of responders (Delta(max) of cortisol >or=9 microg/dl) to non-responders after 1- or 250-microg corticotrophin test did not differ among patients and controls. All patients had a good outcome without glucocorticoid treatment. CONCLUSIONS: Depending on the severity of illness, mild alterations in the HPA axis occurred. However, relative corticosteroid insufficiency was not confirmed in NCIN patients. PMID- 19903799 TI - Clinical relevance of thyroid dysfunction in human haematopoiesis: biochemical and molecular studies. AB - OBJECTIVE: Abnormalities in haematological parameters have been noted in patients with thyroid diseases. Nevertheless, the exact mechanism of thyroid hormones' (THs) action on human haematopoiesis is still not entirely clear. DESIGN: The influence of THs through TH receptors (TRalpha-1 and TRbeta-1) on haematopoiesis in patients with hypo- and hyperthyroidism was analysed. METHODS: TR gene expression at the mRNA and protein levels in human CD34(+)-enriched haematopoietic progenitor cells (HPCs) obtained from the peripheral blood of patients with thyroid disorders and healthy volunteers was analysed. The cell populations were also investigated for clonogenic growth of granulocyte macrophage-colony forming units and erythrocyte-burst forming units (BFU-E). The level of apoptosis was determined by annexin V/propidium iodide staining and quantitative RT-PCR. RESULTS: The studies revealed that hypo- and hyperthyroidism modify TR gene expression in HPCs in vivo. TH deficiency resulted in a decrease in total blood counts and clonogenic potential of BFU-E. In contrast, hyperthyroid patients presented increased clonogenic growth and BFU-E number and significantly higher expressions of cell cycle-regulating genes such as those for PCNA and cyclin D1. Finally, an increase in the frequency of apoptotic CD34(+) enriched HPCs in hypo- and hyperthyroidism with a modulation of apoptosis-related genes was detected. CONCLUSIONS: The following conclusions were derived: i) TR expression in human haematopoietic cells depends on TH status, ii) both hypo- and hyperthyroidism significantly influence clonogenicity and induce apoptosis in CD34(+)-enriched HPCs and iii) the molecular mechanism by which THs influence haematopoiesis might provide a basis for designing novel therapeutic interventions in thyroid diseases. PMID- 19903800 TI - Herpesvirus type 7 infection may play an important role in individuals with a genetic profile of susceptibility to Graves' disease. AB - OBJECTIVE: An inherited profile of genes related to the response to aggressive environmental factors such as viruses and chemicals may be related to an increased susceptibility to Graves' disease (GD). DESIGN AND METHODS: This prospective case-control study was designed to examine the relationship between human herpesviruses (HHV) infection, determined by circulating DNA; tumour protein p53 (TP53) apoptotic ability; and detoxification system genes, and GD. We studied 280 confirmed GD patients paired to 284 controls with respect to environmental exposure. Exclusion criteria included medications that could interfere with thyroid function evaluation and a recent history of viral and bacterial infections. RESULTS: A stepwise regression analysis adjusted for age, gender, and ethnicity established the inheritance of glutathione S-transferase pi 1 (GSTP1) (odds ratio (OR)=3.423; 95% confidence interval (CI)=2.120-5.527; P<0.001) and cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1) variants (OR=1.649; 95% CI=1.012-2.686; P=0.0445) as significant risk factors for the disease. HHV-7 infection was much more common in GD patients (64.64%) than in controls (38.73%; chi(2), P<0.0001), and it increased the risk for GD more than three times (OR=3.133; 95% CI=1.959-5.011; P<0.0001). The inheritance of less efficient Pro/Pro TP53 gene variants significantly increased the risk of GD development (OR=5.196; 95% CI=2.112-12.783; P<0.0001) and also favored HHV-7 infection (OR=2.835; 95% CI=1.100-7.310; P=0.0275). In addition, 72TP53 variants augmented the risk of GD relapse (OR=1.860; 95% CI=1.015-3.410; P=0.0446). CONCLUSIONS: We suggest that an inherited genetic profile involving TP53 may favor HHV-7 infection and maintenance, which, in turn, may initiate and perpetuate GD autoimmune process. PMID- 19903801 TI - Trough serum testosterone predicts the development of polycythemia in hypogonadal men treated for up to 21 years with subcutaneous testosterone pellets. AB - OBJECTIVES: Testosterone formulations that have more steady-state pharmacokinetics, such as subcutaneously implanted testosterone pellets, may cause less erythrocytosis than i.m. injections of shorter acting androgen esters. We, therefore, sought to define the prevalence, predictors, and proximate basis (role of erythropoietin) for polycythemia (hematocrit >0.50) in hypogonadal men receiving testosterone implants long term. DESIGN: A cross-sectional study was conducted in an academic andrology center with a longitudinal subgroup analysis. PATIENTS: A total of 158 hypogonadal men aged 14-84 years (mean age 46.7 years) treated on average for 8 years (range 0-21 years). MEASUREMENTS: Trough blood testosterone and hematocrit. Serial serum erythropoietin concentrations were measured in 16 volunteers. RESULTS: Positive univariate associations between polycythemia (hematocrit >0.50) and log(testosterone) (odds ratio (OR) 24.7, 95% confidence interval (CI): 4.3, 141.2, P<0.01) and age (OR 1.1, 95% CI: 1.0, 1.1, P=0.03) and a borderline relationship with current smoking (OR 4.2, 95% CI: 0.9, 20.0, P=0.08) were unveiled. A sensitivity analysis using alternate definitions of polycythemia was performed to capture all potential covariants. Multivariate regression analysis incorporating all potential covariants disclosed the independent OR of developing polycythemia (after adjusting for smoking and age) for log(testosterone) to be 15.0 (95% CI: 2.5, 90.8). Duration of testosterone therapy did not alter the risk of polycythemia. A direct relationship between testosterone and erythropoietin was observed (P=0.05). CONCLUSIONS: Higher trough serum testosterone concentrations but not duration of treatment predict the development of polycythemia in men receiving long-acting depot testosterone treatment. PMID- 19903802 TI - Lower early morning plasma cortisol levels are associated with thyroid autoimmunity in the elderly. AB - OBJECTIVES: Thyroid autoimmunity decreases in the very old. We investigated whether glucocorticoid (GC) activity, which increases in old age, is involved in this process. SUBJECTS AND METHODS: A total of 321 ambulatory subjects (age 51-95 years, median 71, 207 female) were studied. Thyroid function tests, cortisol, glucose, insulin and biochemical parameters were measured. A modified overnight dexamethasone suppression test (0.25 mg) was performed as an index of GC sensitivity. RESULTS: Forty subjects had positive anti-thyroid peroxidase antibodies and 36 had positive anti-thyroglobulin antibodies, while 57 had either one or the other or both thyroid autoantibodies (ThAbs) positive. Mean basal cortisol levels were significantly lower in the ThAbs (+) groups (320+/-125 vs 378+/-128 nmol/l, P=0.002). Triiodothyronine, free thyroxine, post-dexamethasone cortisol levels, C-reactive protein, homeostasis model assessment-insulin resistance-index and body mass index did not differ between these two groups. Mean age of ThAbs (+) subjects was lower compared to the ThAbs (-) group (67.38+/ 7.38 vs 71.64+/-8.57 years, P=0.001). CONCLUSIONS: Reduced GC activity is associated with an increased prevalence of ThAbs positivity in older ambulatory subjects. Subjects without ThAbs in this population sample are relatively older. It is not known whether this is related to increasing GC activity with age. PMID- 19903803 TI - Re: Discriminatory accuracy from single-nucleotide polymorphisms in models to predict breast cancer risk. PMID- 19903804 TI - Age-specific evaluation of primary human papillomavirus screening vs conventional cytology in a randomized setting. AB - BACKGROUND: Human papillomavirus (HPV) DNA testing has shown higher sensitivity than cytology for detecting cervical lesions, but it is uncertain whether the higher sensitivity is dependent on the age of the woman being screened. We compared the age-specific performance of primary HPV DNA screening with that of conventional cytology screening in the setting of an organized population-based cervical cancer screening program in Finland. METHODS: From January 1, 2003, to December 31, 2005, randomized invitations were sent to women aged 25-65 years for routine cervical cancer screening by primary high-risk HPV DNA testing (n = 54 207) with a Hybrid Capture 2 assay followed by cytology triage for women who were HPV DNA positive or by conventional cytology screening (n = 54 218). In both screening arms, cytology results of low-grade squamous intraepithelial lesion or worse triggered a referral for colposcopy. Relative rates (RRs) of detection to assess test sensitivity, specificity, and positive predictive values (PPVs) with 95% confidence intervals (CIs) were calculated for the histological endpoints of cervical intraepithelial neoplasia (CIN) grade 1 or higher (CIN 1+), CIN grade 2 or higher (CIN 2+), and CIN grade 3 or higher (CIN 3+). All statistical tests were two-sided. RESULTS: The overall frequency of colposcopy referrals was 1.2% in both screening arms. Women younger than 35 years were referred more often in the HPV DNA screening vs the conventional screening arm (RR = 1.27, 95% CI = 1.01 to 1.60). The prevalence of histologically confirmed CIN or cancer was 0.59% in the HPV DNA screening arm vs 0.43% in the conventional screening arm. The relative rates of detection for CIN 1, CIN 2, and CIN 3+ for HPV DNA screening with cytology triage vs conventional screening were 1.44 (95% CI = 0.99 to 2.10), 1.39 (95% CI = 1.03 to 1.88), and 1.22 (95% CI = 0.78 to 1.92), respectively. The specificity of the HPV DNA test with cytology triage was equal to that of conventional screening for all age groups (99.2% vs 99.1% for CIN 2+, P = .13). Among women aged 35 years or older, the HPV DNA test with cytology triage tended to have higher specificity than conventional screening. The PPVs for HPV DNA screening with cytology triage were consistently higher than those for conventional screening. In both screening arms, the test specificities increased with increasing age of the women being screening, whereas the highest PPVs were observed among the youngest women being screened. Overall, 7.2% of women in the HPV DNA screening arm vs 6.6% of women in the conventional screening arm were recommended for intensified follow-up, and the percentages were highest among 25- to 29-year-olds (21.9% vs 10.0%, respectively). CONCLUSIONS: Primary HPV DNA screening with cytology triage is more sensitive than conventional screening. Among women aged 35 years or older, primary HPV DNA screening with cytology triage is also more specific than conventional screening and decreases colposcopy referrals and follow-up tests. PMID- 19903805 TI - Detecting an overall survival benefit that is derived from progression-free survival. AB - BACKGROUND: Whether progression-free survival (PFS) or overall survival (OS) is the more appropriate endpoint in clinical trials of metastatic cancer is controversial. In some disease and treatment settings, an improvement in PFS does not result in an improved OS. METHODS: We partitioned OS into two parts and expressed it as the sum of PFS and survival postprogression (SPP). We simulated randomized clinical trials with two arms that had respective medians for PFS of 6 and 9 months. We assumed no treatment difference in median SPP. We found the probability of a statistically significant benefit in OS for various median SPP and observed P values for PFS. We compared the sample sizes required for PFS vs OS for various median SPP. We compare our results with the literature regarding surrogacy of PFS for OS by use of the correlation between hazard ratios for PFS and OS. All statistical tests were two-sided. RESULTS: For a trial with observed P value for improvement in PFS of .001, there was a greater than 90% probability for statistical significance in OS if median SPP was 2 months but less than 20% if median SPP was 24 months. For a trial requiring 280 patients to detect a 3 month difference in PFS, 350 and 2440 patients, respectively, were required to have the same power for detecting a real difference in OS that is carried over from the 3-month benefit in PFS when the median SPP was 2 and 24 months. CONCLUSIONS: Addressing SPP is important in understanding treatment effects. For clinical trials with a PFS benefit, lack of statistical significance in OS does not imply lack of improvement in OS, especially for diseases with long median SPP. Although there may be no treatment effect on SPP, its variability so dilutes the OS comparison that statistical significance is likely lost. OS is a reasonable primary endpoint when median SPP is short but is too high a bar when median SPP is long, such as longer than 12 months. PMID- 19903806 TI - A new generation of studies of human papillomavirus DNA testing in cervical cancer screening. PMID- 19903807 TI - Antitumor activity of the selective MDM2 antagonist nutlin-3 against chemoresistant neuroblastoma with wild-type p53. AB - BACKGROUND: Restoring p53 function by antagonizing its interaction with the negative regulator MDM2 is an appealing nongenotoxic approach to treating tumors with wild-type p53. Mutational inactivation of p53 is rare in neuroblastoma tumors at diagnosis and occurs in only a subset of multidrug-resistant neuroblastomas. METHODS: The antiproliferative and cytotoxic effect of nutlin-3, a small-molecule MDM2 antagonist, was examined in chemosensitive (UKF-NB-3) and matched chemoresistant neuroblastoma cells with wild-type p53 (UKF-NB-3(r)DOX20) or with mutant p53 (UKF-NB-3(r)VCR10). Activation of the p53 pathway was assessed by expression analysis of p53 target genes, flow cytometric cell cycle analysis, and apoptosis assays. Mice with established chemoresistant tumor xenografts were treated orally with nutlin-3 or vehicle control (n = 5-10 mice per group) and were used to evaluate effects on tumor growth, p53 pathway activity, and metastatic tumor burden. All statistical tests were two-sided. RESULTS: Nutlin-3 induced a similar activation of the p53 pathway in UKF-NB-3 and UKF-NB-3(r)DOX20 cells, as evidenced by increased expression of p53 target genes, G1 cell cycle arrest, and induction of apoptosis. No such response was observed in UKF-NB 3(r)VCR10 cells with mutant p53. Oral administration of nutlin-3 to UKF-NB 3(r)DOX20 xenograft-bearing mice led to inhibition of primary tumor growth (mean tumor volume after 3 weeks of treatment, nutlin-3- vs vehicle-treated mice: 772 vs 1661 mm3, difference = 890 mm3, 95% confidence interval = 469 to 1311 mm3, P < .001), p53 pathway activation, and reduction in the extent of metastatic disease. The growth of UKF-NB-3(r)VCR10 xenografts was unaffected by nutlin-3. CONCLUSIONS: Nutlin-3 activates the p53 pathway and suppresses tumor growth in this model system of chemoresistant neuroblastoma, provided that wild-type p53 is present. PMID- 19903808 TI - Patterns of use and risks associated with erythropoiesis-stimulating agents among Medicare patients with cancer. AB - BACKGROUND: Erythropoiesis-stimulating agents (erythropoietin and darbepoietin) have been approved to reduce the number of blood transfusions required during chemotherapy; however, concerns about the risks of venous thromboembolism and mortality exist. METHODS: We identified patients who were aged 65 years or older in the Surveillance, Epidemiology, and End Results-Medicare database; who were diagnosed with colon, non-small cell lung, or breast cancer or with diffuse large B-cell lymphoma from January 1, 1991, through December 31, 2002; and who received chemotherapy. The main outcome measures were claims for use of an erythropoiesis stimulating agent, blood transfusion, venous thromboembolism (ie, deep vein thrombosis or pulmonary embolism), and overall survival. We used multivariable logistic regression models to analyze the association of erythropoiesis stimulating agent use with clinical and demographic variables. We used time dependent Cox proportional hazards models to analyze the association of time to receipt of first erythropoiesis-stimulating agent with venous thromboembolism and overall survival. All statistical tests were two-sided. RESULTS: Among 56,210 patients treated with chemotherapy from 1991 through 2002, 15,346 (27%) received an erythropoiesis-stimulating agent. The proportion of patients receiving erythropoiesis-stimulating agents increased from 4.8% in 1991 to 45.9% in 2002 (P < .001). Use was associated with more recent diagnosis, younger age, urban residence, comorbidities, receipt of radiation therapy, female sex, and metastatic or recurrent cancer. The rate of blood transfusion per year during 1991-2002 remained constant at 22%. Venous thromboembolism developed in 1796 (14.3%) of the 12,522 patients who received erythropoiesis-stimulating agent and 3400 (9.8%) of the 34,820 patients who did not (hazard ratio = 1.93, 95% confidence interval = 1.79 to 2.07). Overall survival was similar in both groups. CONCLUSION: Use of erythropoiesis-stimulating agent increased rapidly after its approval in 1991, but the blood transfusion rate did not change. Use of erythropoiesis-stimulating agents was associated with an increased risk of venous thromboembolism but not of mortality. PMID- 19903809 TI - Targeted molecular therapy for neuroblastoma: the ARF/MDM2/p53 axis. PMID- 19903810 TI - Kidins220/ARMS modulates the activity of microtubule-regulating proteins and controls neuronal polarity and development. AB - In order for neurons to perform their function, they must establish a highly polarized morphology characterized, in most of the cases, by a single axon and multiple dendrites. Herein we find that the evolutionarily conserved protein Kidins220 (kinase D-interacting substrate of 220-kDa), also known as ARMS (ankyrin repeat-rich membrane spanning), a downstream effector of protein kinase D and neurotrophin and ephrin receptors, regulates the establishment of neuronal polarity and development of dendrites. Kidins220/ARMS gain and loss of function experiments render severe phenotypic changes in the processes extended by hippocampal neurons in culture. Although Kidins220/ARMS early overexpression hinders neuronal development, its down-regulation by RNA interference results in the appearance of multiple longer axon-like extensions as well as aberrant dendritic arbors. We also find that Kidins220/ARMS interacts with tubulin and microtubule-regulating molecules whose role in neuronal morphogenesis is well established (microtubule-associated proteins 1b, 1a, and 2 and two members of the stathmin family). Importantly, neurons where Kidins220/ARMS has been knocked down register changes in the phosphorylation activity of MAP1b and stathmins. Altogether, our results indicate that Kidins220/ARMS is a key modulator of the activity of microtubule-regulating proteins known to actively regulate neuronal morphogenesis and suggest a mechanism by which it contributes to control neuronal development. PMID- 19903811 TI - C-reactive protein exists in an NaCl concentration-dependent pentamer-decamer equilibrium in physiological buffer. AB - C-reactive protein (CRP) is an acute phase protein of the pentraxin family that binds ligands in a Ca(2+)-dependent manner, and activates complement. Knowledge of its oligomeric state in solution and at surfaces is essential for functional studies. Analytical ultracentrifugation showed that CRP in 2 mM Ca(2+) exhibits a rapid pentamer-decamer equilibrium. The proportion of decamer decreased with an increase in NaCl concentration. The sedimentation coefficients s(20,w)(0) of pentameric and decameric CRP were 6.4 S and in excess of 7.6 S, respectively. In the absence of Ca(2+), CRP partially dissociates into its protomers and the NaCl concentration dependence of the pentamer-decamer equilibrium is much reduced. By x-ray scattering, the radius of gyration R(G) values ranged from 3.7 nm for the pentamer to above 4.0 nm for the decamer. An averaged K(D) value of 21 microM in solution (140 mM NaCl, 2 mM Ca(2+)) was determined by x-ray scattering and modeling based on crystal structures for the pentamer and decamer. Surface plasmon resonance showed that CRP self-associates on a surface with immobilized CRP with a similar K(D) value of 23 microM (140 mM NaCl, 2 mM Ca(2+)), whereas CRP aggregates in low salt. It is concluded that CRP is reproducibly observed in a pentamer-decamer equilibrium in physiologically relevant concentrations both in solution and on surfaces. Both 2 mM Ca(2+) and 140 mM NaCl are essential for the integrity of CRP in functional studies and understanding the role of CRP in the acute phase response. PMID- 19903812 TI - Proline is not uniquely capable of providing the pivot point for domain swapping in 2G12, a broadly neutralizing antibody against HIV-1. AB - The human monoclonal antibody 2G12 is a member of a small group of broadly neutralizing antibodies against human immunodeficiency virus type 1. 2G12 adopts a unique variable heavy domain-exchanged dimeric configuration that results in an extensive multivalent binding surface and the ability to bind with high affinity to densely clustered high mannose oligosaccharides on the "silent" face of the gp120 envelope glycoprotein. Here, we further define the amino acids responsible for this extraordinary domain-swapping event in 2G12. PMID- 19903813 TI - NFkappaB p50-CCAAT/enhancer-binding protein beta (C/EBPbeta)-mediated transcriptional repression of microRNA let-7i following microbial infection. AB - MicroRNAs, central players of numerous cellular processes, regulate mRNA stability or translational efficiency. Although these molecular events are established, the mechanisms regulating microRNA function and expression remain largely unknown. The microRNA let-7i regulates Toll-like receptor 4 expression. Here, we identify a novel transcriptional mechanism induced by the protozoan parasite Cryptosporidium parvum and Gram(-) bacteria-derived lipopolysaccharide (LPS) mediating let-7i promoter silencing in human biliary epithelial cells (cholangiocytes). Using cultured cholangiocytes, we show that microbial stimulus decreased let-7i expression, and promoter activity. Analysis of the mechanism revealed that microbial infection promotes the formation of a NFkappaB p50 C/EBPbeta silencer complex in the regulatory sequence. Chromatin immunoprecipitation assays (ChIP) demonstrated that the repressor complex binds to the let-7i promoter following microbial stimulus and promotes histone-H3 deacetylation. Our results provide a novel mechanism of transcriptional regulation of cholangiocyte let-7i expression following microbial insult, a process with potential implications for epithelial innate immune responses in general. PMID- 19903815 TI - PB1 domain interaction of p62/sequestosome 1 and MEKK3 regulates NF-kappaB activation. AB - p62/Sequestosome 1 is a scaffold protein involved in the regulation of autophagy, trafficking of proteins to the proteasome, and activation of NF-kappaB. p62 encodes an N-terminal PB1 domain in addition to the ZZ domain, TRAF6-binding domain, LC3 interaction region, and ubiquitin-associated domain, each critical for the physiological function of p62. PB1 domains have a beta-grasp topology where the front end of one PB1 domain binds the back end of a second PB1 domain. The p62 PB1 domain homodimerizes as well as heterodimerizes with other PB1 domains. The front end of the PB1 domain in p62 binds the PB1 domain of atypical protein kinases C, the MAPK kinase, MEK5, and the NBR1 protein. Other than its role in homodimerization, the rear end acidic cluster region of the p62 PB1 domain had no previous defined binding partners. Herein, we demonstrate that the rear end acidic cluster region of the p62 PB1 domain binds the front end basic region of the MAPK kinase kinase, MEKK3. p62 and MEKK3 co-localize in speckles or aggregates that are centers for organizing TRAF6-regulated NF-kappaB signaling and the assembly of polyubiquinated proteins sorting to sequestosomes and proteasomes. The p62-MEKK3 complex binds TRAF6, which regulates the ubiquitination of the IKK complex and NF-kappaB activation. p62 is required for the association of MEKK3 with TRAF6 and short hairpin RNA knockdown of p62 inhibits IL-1 and MEKK3 activation of NF-kappaB. The rear end acidic cluster of the p62 PB1 domain is used to organize cytosolic aggregates or speckles associated TRAF6-p62-MEKK3 complex for control of NF-kappaB activation. PMID- 19903814 TI - Farnesyl pyrophosphate inhibits epithelialization and wound healing through the glucocorticoid receptor. AB - Farnesyl pyrophosphate (FPP), a key intermediate in the mevalonate pathway and protein farnesylation, can act as an agonist for several nuclear hormone receptors. Here we show a novel mechanism by which FPP inhibits wound healing acting as an agonist for glucocorticoid receptor (GR). Elevation of endogenous FPP by the squalene synthetase inhibitor zaragozic acid A (ZGA) or addition of FPP to the cell culture medium results in activation and nuclear translocation of the GR, a known wound healing inhibitor. We used functional studies to evaluate the effects of FPP on wound healing. Both FPP and ZGA inhibited keratinocyte migration and epithelialization in vitro and ex vivo. These effects were independent of farnesylation and indicate that modulation of FPP levels in skin may be beneficial for wound healing. FPP inhibition of keratinocyte migration and wound healing proceeds, in part, by repression of the keratin 6 gene. Furthermore, we show that the 3-hydroxy-3-methylglutaryl-CoA-reductase inhibitor mevastatin, which blocks FPP formation, not only promotes epithelialization in acute wounds but also reverses the effect of ZGA on activation of the GR and inhibition of epithelialization. We conclude that FPP inhibits wound healing by acting as a GR agonist. Of special interest is that FPP is naturally present in cells prior to glucocorticoid synthesis and that FPP levels can be further altered by the statins. Therefore, our findings may provide a better understanding of the pleiotropic effects of statins as well as molecular mechanisms by which they may accelerate wound healing. PMID- 19903816 TI - A biochemical and functional protein complex involving dopamine synthesis and transport into synaptic vesicles. AB - Synaptic transmission depends on neurotransmitter pools stored within vesicles that undergo regulated exocytosis. In the brain, the vesicular monoamine transporter-2 (VMAT(2)) is responsible for the loading of dopamine (DA) and other monoamines into synaptic vesicles. Prior to storage within vesicles, DA synthesis occurs at the synaptic terminal in a two-step enzymatic process. First, the rate limiting enzyme tyrosine hydroxylase (TH) converts tyrosine to di-OH phenylalanine. Aromatic amino acid decarboxylase (AADC) then converts di-OH phenylalanine into DA. Here, we provide evidence that VMAT(2) physically and functionally interacts with the enzymes responsible for DA synthesis. In rat striata, TH and AADC co-immunoprecipitate with VMAT(2), whereas in PC 12 cells, TH co-immunoprecipitates with the closely related VMAT(1) and with overexpressed VMAT(2). GST pull-down assays further identified three cytosolic domains of VMAT(2) involved in the interaction with TH and AADC. Furthermore, in vitro binding assays demonstrated that TH directly interacts with VMAT(2). Additionally, using fractionation and immunoisolation approaches, we demonstrate that TH and AADC associate with VMAT(2)-containing synaptic vesicles from rat brain. These vesicles exhibited specific TH activity. Finally, the coupling between synthesis and transport of DA into vesicles was impaired in the presence of fragments involved in the VMAT(2)/TH/AADC interaction. Taken together, our results indicate that DA synthesis can occur at the synaptic vesicle membrane, where it is physically and functionally coupled to VMAT(2)-mediated transport into vesicles. PMID- 19903817 TI - Clostridium perfringens enterotoxin interacts with claudins via electrostatic attraction. AB - Clostridium perfringens enterotoxin (CPE), a causative agent of food poisoning, is a pore-forming toxin disrupting the selective permeability of the plasma membrane of target cells, resulting in cell death. We previously identified claudin as the cell surface receptor for CPE. Claudin, a component of tight junctions, is a tetratransmembrane protein and constitutes a large family of more than 20 members, not all of which serve as the receptor for CPE. The mechanism by which the toxin distinguishes the sensitive claudins is unknown. In this study, we localized the region of claudin responsible for interaction with CPE to the C terminal part of the second extracellular loop and found that the isoelectric point of this region in sensitive claudins was higher than insensitive claudins. Amino acid substitutions to lower the pI resulted in reduced sensitivity to CPE among sensitive claudins, whereas substitutions to raise the pI endowed CPE insensitive claudins with sensitivity. The steric structure of the claudin binding domain of CPE reveals an acidic cleft surrounded by Tyr(306), Tyr(310), Tyr(312), and Leu(315), which were reported to be essential for interaction with the sensitive claudins. These results imply that an electrostatic attraction between the basic claudin region and the acidic CPE cleft is involved in their interaction. PMID- 19903818 TI - Functional analysis of the Kv1.1 N255D mutation associated with autosomal dominant hypomagnesemia. AB - Mutations in the voltage-gated K(+) channel Kv1.1 have been linked with a mixed phenotype of episodic ataxia and/or myokymia. Recently, we presented autosomal dominant hypomagnesemia as a new phenotypic characteristic associated with a mutation in Kv1.1 (N255D) (Glaudemans, B., van der Wijst, J., Scola, R. H., Lorenzoni, P. J., Heister, A., van der Kemp, A. W., Knoers, N. V., Hoenderop, J. G., and Bindels, R. J. (2009) J. Clin. Invest. 119, 936-942). A conserved asparagine at position 255 in the third transmembrane segment was converted into an aspartic acid, resulting in a non-functional channel. In this study, we explored the functional consequence of this conserved residue by substitution with other hydrophobic, polar, or charged amino acids (N255E, N255Q, N255A, N255V, N255T, and N255H). Upon overexpression in human embryonic kidney (HEK293) cells, cell surface biotinylation revealed plasma membrane expression of all mutant channels. Next, we used the whole-cell patch clamp technique to demonstrate that the N255E and N255Q mutants were non-functional. Substitution of Asn-255 with other amino acids (N255A, N255V, N255T, and N255H) did not prevent ion conduction, and these mutant channels activated at more negative potentials when compared with wild-type channels, -41.5 +/- 1.6, -45.5 +/- 2.0, -50.5 +/- 1.9, and -33.8 +/- 1.3 mV to -29.4 +/- 1.1 mV, respectively. The time constant of activation was significantly faster for the two most hydrophobic mutations, N255A (6.2 +/- 0.2 ms) and N255V (5.2 +/- 0.3 ms), and the hydrophilic mutant N255T (9.8 +/- 0.4 ms) in comparison with wild type (13.0 +/- 0.9 ms). Furthermore, the voltage dependence of inactivation was shifted approximately 13 mV to more negative potentials in all mutant channels except for N255H. Taken together, our data showed that an asparagine at position 255 in Kv1.1 is required for normal voltage dependence and kinetics of channel gating. PMID- 19903819 TI - Isolation of novel animal cell lines defective in glycerolipid biosynthesis reveals mutations in glucose-6-phosphate isomerase. AB - Glycerolipids are structural components for membranes and serve in energy storage. We describe here the use of a photodynamic selection technique to generate a population of Chinese hamster ovary cells that display a global deficiency in glycerolipid biosynthesis. One isolate from this population, GroD1, displayed a profound reduction in the synthesis of phosphatidylcholine, phosphatidylethanolamine, and triglycerides but presented high levels of phosphatidic acid and normal levels of phosphatidylinositol synthesis. This was accompanied by a reduction in phosphatidate phosphatase 1 (PAP1) activity. Expression cloning and sequencing of the cDNA obtained from GroD1 revealed a point mutation, Gly-189 --> Glu, in glucose-6-phosphate isomerase (GPI), a glycolytic enzyme involved in an inherited disorder that results in anemia and neuromuscular symptoms in humans. GPI activity was reduced by 87% in GroD1. No significant differences were found in DNA synthesis, protein synthesis, and ATP levels, whereas glycerol 3-phosphate levels were increased in the mutant. Expression of wild-type hamster GPI restored GPI activity, glycerolipid biosynthesis, and PAP1 activity in GroD1. Two additional, independently isolated GPI-deficient mutants displayed similar phenotypes with respect to PAP1 activity and glycerolipid biosynthesis. These findings uncover a novel relationship between GPI, involved in carbohydrate metabolism, and PAP1, a lipogenic enzyme. These results may also help to explain neuromuscular symptoms associated with inherited GPI deficiency. PMID- 19903820 TI - Molecular basis for association of PIPKI gamma-p90 with clathrin adaptor AP-2. AB - Phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)) is an essential determinant in clathrin-mediated endocytosis (CME). In mammals three type I phosphatidylinositol-4-phosphate 5-kinase (PIPK) enzymes are expressed, with the I gamma-p90 isoform being highly expressed in the brain where it regulates synaptic vesicle (SV) exo-/endocytosis at nerve terminals. How precisely PI(4,5)P(2) metabolism is controlled spatially and temporally is still uncertain, but recent data indicate that direct interactions between type I PIPK and components of the endocytic machinery, in particular the AP-2 adaptor complex, are involved. Here we demonstrated that PIPKI gamma-p90 associates with both the mu and beta2 subunits of AP-2 via multiple sites. Crystallographic data show that a peptide derived from the splice insert of the human PIPKI gamma-p90 tail binds to a cognate recognition site on the sandwich subdomain of the beta2 appendage. Partly overlapping aromatic and hydrophobic residues within the same peptide also can engage the C-terminal sorting signal binding domain of AP-2mu, thereby potentially competing with the sorting of conventional YXXO motif-containing cargo. Biochemical and structure-based mutagenesis analysis revealed that association of the tail domain of PIPKI gamma-p90 with AP-2 involves both of these sites. Accordingly the ability of overexpressed PIPKI gamma tail to impair endocytosis of SVs in primary neurons largely depends on its association with AP 2 beta and AP-2mu. Our data also suggest that interactions between AP-2 and the tail domain of PIPKI gamma-p90 may serve to regulate complex formation and enzymatic activity. We postulate a model according to which multiple interactions between PIPKI gamma-p90 and AP-2 lead to spatiotemporally controlled PI(4,5)P(2) synthesis during clathrin-mediated SV endocytosis. PMID- 19903821 TI - N terminus is key to the dominant negative suppression of Ca(V)2 calcium channels: implications for episodic ataxia type 2. AB - Expression of the calcium channels Ca(V)2.1 and Ca(V)2.2 is markedly suppressed by co-expression with truncated constructs containing Domain I. This is the basis for the phenomenon of dominant negative suppression observed for many of the episodic ataxia type 2 mutations in Ca(V)2.1 that predict truncated channels. The process of dominant negative suppression has been shown previously to stem from interaction between the full-length and truncated channels and to result in downstream consequences of the unfolded protein response and endoplasmic reticulum-associated protein degradation. We have now identified the specific domain that triggers this effect. For both Ca(V)2.1 and Ca(V)2.2, the minimum construct producing suppression was the cytoplasmic N terminus. Suppression was enhanced by tethering the N terminus to the membrane with a CAAX motif. The 11 amino acid motif (including Arg(52) and Arg(54)) within the N terminus, which we have previously shown to be required for G protein modulation, is also essential for dominant negative suppression. Suppression is prevented by addition of an N terminal tag (XFP) to the full-length and truncated constructs. We further show that suppression of Ca(V)2.2 currents by the N terminus-CAAX construct is accompanied by a reduction in Ca(V)2.2 protein level, and this is also prevented by mutation of Arg(52) and Arg(54) to Ala in the truncated construct. Taken together, our evidence indicates that both the extreme N terminus and the Arg(52), Arg(54) motif are involved in the processes underlying dominant negative suppression. PMID- 19903822 TI - Activation of calcitonin receptor and calcitonin receptor-like receptor by membrane-anchored ligands. AB - G protein-coupled receptors (GPCRs) are the most important pharmaceutical targets, and more than 40% of drugs in use today modulate GPCR signaling. A major hurdle in the development of therapies targeting GPCRs is the drug candidate's nonselective actions in multiple tissues. The ability to spatially control GPCR signaling would provide a venue for developing therapies that require targeted GPCR signaling. Here, we show that the fusion of a RAMP1 co-receptor with the calcitonin gene-related peptide (CGRP), or calcitonin, transforms the RAMP1 from a co-receptor to bona fide membrane-anchored ligands (CGRP-RAMP1 and CAL-RAMP1). The CAL-RAMP1 selectively activates the calcitonin receptor (CR), whereas, the CGRP-RAMP1 activates both the calcitonin receptor-like receptor (CLR) and CR. Unlike a free peptide, which moves freely in the extracellular space and differentiates targets based on molecular affinity, the anchored CGRP-RAMP1 and CAL-RAMP1 ligands confine their activities to individual cells. In addition, our study showed that a CGRP8-37-RAMP1 chimera, but not RAMP1, functions as an antagonist for CGRP-RAMP1-mediated signaling, suggesting that the activation of CLR by CGRP-RAMP1 shares similar molecular mechanisms with the CGRP-mediated activation of CLR/RAMP1 receptor complexes. Taken together, our finding thus provides a novel class of ligands that activate CR and CLR exclusively in an autocrine manner and a proof-of-concept demonstration for future development of targeted therapies aimed at these receptors in specific cell populations. PMID- 19903824 TI - Evidence for ligand-mediated selective modulation of aryl hydrocarbon receptor activity. AB - The concept of selective receptor modulators has been established for the nuclear steroid hormone receptors. Such selective modulators have been used therapeutically with great success in the treatment of cancer. However, this concept has not been examined with regard to the aryl hydrocarbon receptor (AHR) because of the latent toxicity commonly associated with AHR activation. AHR mediated toxicity is primarily derived from AHR binding to its dioxin response element (DRE) and driving expression of CYP1 family members, which have the capacity to metabolize procarcinogens to genotoxic carcinogens. Recent evidence using a non-DRE binding AHR mutant has established the DRE-independent suppression of inflammatory markers by the AHR. We wished to determine whether such DRE-independent repression with wild-type AHR could be dissociated from canonical DRE-dependent transactivation in a ligand-dependent manner and, in doing so, prove the concept of a selective AHR modulator (SAhRM). Here, we identify the selective estrogen receptor (ER) modulator Way-169916 as a dually selective modulator, binding both ER and AHR. Inflammatory gene expression associated with the cytokine-inducible acute-phase response (e.g., SAA1 and CRP) are diminished by Way-169916 in an AHR-dependent manner. Furthermore, activation of AHR by Way-169916 fails to stimulate canonical DRE-driven AHR-mediated CYP1A1 expression, thus eliminating the potential for AHR-mediated genotoxic stress. Such anti-inflammatory activity in the absence of DRE-mediated expression fulfills the major criteria of an SAhRM, which suggests that selective modulation of AHR is possible and renders the AHR a therapeutically viable drug target for the amelioration of inflammatory disease. PMID- 19903823 TI - Large T antigen promotes JC virus replication in G2-arrested cells by inducing ATM- and ATR-mediated G2 checkpoint signaling. AB - Large T antigen (TAg) of the human polyomavirus JC virus (JCV) possesses DNA binding and helicase activities, which, together with various cellular proteins, are required for replication of the viral genome. We now show that JCV-infected cells expressing TAg accumulate in the G(2) phase of the cell cycle as a result of the activation of ATM- and ATR-mediated G(2) checkpoint pathways. Transient transfection of cells with a TAg expression vector also induced G(2) checkpoint signaling and G(2) arrest. Analysis of TAg mutants with different subnuclear localizations suggested that the association of TAg with cellular DNA contributes to the induction of G(2) arrest. Abrogation of G(2) arrest by inhibition of ATM and ATR, Chk1, and Wee1 suppressed JCV genome replication. In addition, abrogation of the G(2)-M transition by Cdc2 depletion disabled Wee1 depletion induced suppression of JCV genome replication, suggesting that JCV replication is facilitated by G(2) arrest resulting from G(2) checkpoint signaling. Moreover, inhibition of ATM and ATR by caffeine suppressed JCV production. The observation that oligodendrocytes productively infected with JCV in vivo also undergo G(2) arrest suggests that G(2) checkpoint inhibitors such as caffeine are potential therapeutic agents for JCV infection. PMID- 19903825 TI - Identification of dynamin-2-mediated endocytosis as a new target of osteoporosis drugs, bisphosphonates. AB - Nitrogen-containing bisphosphonates are pyrophosphate analogs that have long been the preferred prescription for treating osteoporosis. Although these drugs are considered inhibitors of prenylation and are believed to exert their effects on bone resorption by disrupting the signaling pathways downstream of prenylated small GTPases, this explanation seems to be insufficient. Because other classes of prenylation inhibitors have recently emerged as potential antiviral therapeutic agents, we first investigated here the effects of bisphosphonates on simian virus 40 and adenovirus infections and, to our surprise, found that viral infections are suppressed by bisphosphonates through a prenylation-independent pathway. By in-house affinity-capture techniques, dynamin-2 was identified as a new molecular target of bisphosphonates. We present evidence that certain bisphosphonates block endocytosis of adenovirus and a model substrate by inhibiting GTPase activity of dynamin-2. Hence, this study has uncovered a previously unknown mechanism of action of bisphosphonates and offers potential novel use for these drugs. PMID- 19903826 TI - A novel roscovitine derivative potently induces G1-phase arrest in platelet derived growth factor-BB-activated vascular smooth muscle cells. AB - Abnormal vascular smooth muscle cell (VSMC) proliferation contributes to the pathogenesis of restenosis. Thus, drugs interfering with cell cycle progression in VSMC are promising candidates for an antirestenotic therapy. In this study, we pharmacologically characterize N-5-(2-aminocyclohexyl)-N-7-benzyl-3-isopropyl 1(2)H-pyrazolo[4,3-d]pyrimidine-5,7-di-amine (LGR1406), a novel derivative of the cyclin-dependent kinase (CDK) inhibitor roscovitine (ROSC), in PDGF-BB-activated VSMC. Cell proliferation was quantified measuring DNA synthesis via 5-bromo-2' deoxyuridine incorporation. Analysis of cell cycle distribution was done by flow cytometry using propidium iodide-stained nuclei. Key regulators of the cell cycle and relevant signaling pathways were dissected by Western blot analyses. In addition, in vitro kinase assays and in silico studies regarding the pharmacokinetic profile of both compounds were performed. LGR1406 shows a stronger (IC(50) = 3.0 muM) antiproliferative activity than ROSC (IC(50) = 16.9 muM), halting VSMCs in G(0)/G(1) phase of the cell cycle, whereas ROSC does not arrest but rather delays cell cycle progression. Neither of the compounds interferes with early PDGF-BB-induced signaling pathways (p38, extracellular signal-regulated kinase 1/2, c-Jun NH(2)-terminal kinase, Akt, signal transducer and activator of transcription 3), and both inhibit CDKs, with LGR1406 exerting a slightly higher potency against CDK1/2 and 4 than ROSC. Expression of cyclins A and E as well as hyperphosphorylation of the pocket proteins retinoblastoma protein and p107 are negatively affected by both compounds, although to a different extent. In silico calculations predicted a much higher metabolic stability for LGR1406 compared with ROSC. Altogether, ROSC derivatives, such as LGR1406 seem to be promising compounds for further development in antirestenotic therapy. PMID- 19903827 TI - G protein-mediated inhibition of Cav3.2 T-type channels revisited. AB - T-type calcium channels are important modulators of both membrane potential and intracellular Ca(2+) concentration, allowing them to play key roles in such diverse processes as aldosterone production from adrenal glomerulosa cells to boosting pain signals in nociceptors. In both these examples, the Ca(v)3.2 isoform mediates Ca(2+) influx. This isoform is also of particular interest because mutations in its gene (CACNA1H) that enhance channel activity have been associated with idiopathic generalized epilepsies, whereas mutations that disrupt its activity have been associated with autism spectrum disorders. Block of T channel activity has been proposed to contribute to the therapeutic usefulness of a wide variety of drugs, such as antihypertensives, antipsychotics, and antidepressants. Recent evidence strongly supports the hypothesis that block of Ca(v)3.2 channels might be useful in the treatment of neuropathic pain. Therefore, it is of particular interest that Ca(v)3.2 channels are exquisitely regulated by G protein-coupled receptors and various downstream effectors. This Perspective summarizes recent findings (p. 202) on this regulation and the novel pathways specifically activated by either neurokinin I, corticotropin-releasing factor receptor 1, or dopamine D(1) receptors. PMID- 19903828 TI - Dependence of multidrug resistance protein-mediated cyclic nucleotide efflux on the background sodium conductance. AB - Anterior pituitary cells fire action potentials and release cyclic nucleotides both spontaneously and in response to agonist stimulation, but the relationship between electrical activity and cyclic nucleotide efflux has not been studied. In these cells, a tetrodotoxin-resistant background N(+) conductance is critical for firing of action potentials, and multidrug resistance proteins (MRPs) MRP4 and MRP5 contribute to cyclic nucleotide efflux. Here, we show that abolition of the background Na(+) conductance in rat pituitary cells by complete or partial replacement of extracellular Na(+) with organic cations or sucrose induced a rapid and reversible hyperpolarization of cell membranes and inhibition of action potential firing, accompanied by a rapid inhibition of cyclic nucleotide efflux. Valinomycin-induced hyperpolarization of plasma membranes also inhibited cyclic nucleotide efflux, whereas depolarization of cell membranes induced by the inhibition of Ca(2+) influx or stimulation of Na(+) influx by gramicidin was accompanied by a facilitation of cyclic nucleotide efflux. In contrast, inhibition of cyclic nucleotide efflux by probenecid did not affect the background Na(+) conductance. In human embryonic kidney 293 cells stably transfected with human MRP4 or MRP5, replacement of bath Na(+) with organic cations also hyperpolarized the cell membranes and inhibited cyclic nucleotide efflux. In these cells, the Na(+)/H(+) antiporter monensin did not affect the membrane potential and was practically ineffective in altering cyclic nucleotide efflux. In both pituitary and MRP4- and MRP5-expressing cells, 3-[[3-[2-(7 chloroquinolin-2-yl)vinyl]phenyl]-(2 dimethylcarbamoylethylsulfanyl)methylsulfanyl] propionic acid (MK571) inhibited cyclic nucleotide efflux. These results indicate that the MRP4/5-mediated cyclic nucleotide efflux can be rapidly modulated by membrane potential determined by the background Na(+) conductance. PMID- 19903829 TI - A novel multilevel statistical method for the study of the relationships between multireceptorial binding affinity profiles and in vivo endpoints. AB - The present work introduces a novel method for drug research based on the sequential building of linked multivariate statistical models, each one introducing a different level of drug description. The use of multivariate methods allows us to overcome the traditional one-target assumption and to link in vivo endpoints with drug binding profiles, involving multiple receptors. The method starts with a set of drugs, for which in vivo or clinical observations and binding affinities for potentially relevant receptors are known, and allows obtaining predictions of the in vivo endpoints highlighting the most influential receptors. Moreover, provided that the structure of the receptor binding sites is known (experimentally or by homology modeling), the proposed method also highlights receptor regions and ligand-receptor interactions that are more likely to be linked to the in vivo endpoints, which is information of high interest for the design of novel compounds. The method is illustrated by a practical application dealing with the study of the metabolic side effects of antipsychotic drugs. Herein, the method detects related receptors confirmed by experimental results. Moreover, the use of structural models of the receptor binding sites allows identifying regions and ligand-receptor interactions that are involved in the discrimination between antipsychotic drugs that show metabolic side effects and those that do not. The structural results suggest that the topology of a hydrophobic sandwich involving residues in transmembrane helices (TM) 3, 5, and 6, as well as the assembling of polar residues in TM5, are important discriminators between target/antitarget receptors. Ultimately, this will provide useful information for the design of safer compounds inducing fewer side effects. PMID- 19903830 TI - Identification of novel small-molecule agonists for human formyl peptide receptors and pharmacophore models of their recognition. AB - N-formyl peptide receptor (FPR1) and N-formyl peptide receptor-like 1 (FPRL1, now known as FPR2) are G protein-coupled receptors involved in host defense and sensing cellular dysfunction. Because of the potential for FPR1/FPR2 as a therapeutic target, our recent high-throughput screening efforts have focused on the identification of unique nonpeptide agonists of FPR1/FPR2. In the present studies, we screened a chemolibrary of drug-like molecules for their ability to induce intracellular calcium mobilization in RBL-2H3 cells transfected with human FPR1 or FPR2. Screening of these compounds resulted in the identification of novel and potent agonists that activated both FPR1 and FPR2, as well as compounds that were specific for either FPR1 or FPR2 with EC(50) values in the low micromolar range. Specificity of the compounds was supported by analysis of calcium mobilization in HL-60 cells transfected with human FPR1 and FPR2. In addition, all but one agonist activated intracellular calcium flux and chemotaxis in human neutrophils, irrespective of agonist specificity for FPR1 or FPR2. Molecular modeling of the group of FPR1 and FPR2 agonists using field point methodology allowed us to create pharmacophore models for ligand binding sites and formulate requirements for these specific N-formyl peptide receptor agonists. These studies further demonstrate that agonists of FPR1/FPR2 include compounds with wide chemical diversity and that analysis of such compounds can enhance our understanding of their ligand/receptor interaction. PMID- 19903831 TI - An inhibitor of protein arginine methyltransferases, 7,7' carbonylbis(azanediyl)bis(4-hydroxynaphthalene-2-sulfonic acid (AMI-1), is a potent scavenger of NADPH-oxidase-derived superoxide. AB - The methylation of proteins is an important post-translational mechanism that has been established to influence the activity of nuclear and nucleic acid binding proteins. Much less is known about the importance of protein methylation in the regulation of cytosolic proteins. Increased methylation of proteins is observed in cardiovascular disease and occurs in conjunction with elevated production of reactive oxygen species. However, the nature of the relationship between reactive oxygen species and protein methylation is poorly understood. Therefore, the goal of the current study was to determine whether protein methylation influences the catalytic activity of the NADPH oxidases (Nox), which are a family of enzymes responsible for the generation of superoxide. We found that the selective inhibitor of protein arginine methyltransferases 7,7'-carbonylbis(azanediyl)bis(4 hydroxynaphthalene-2-sulfonic acid (AMI-1) was a potent antagonist of Nox-derived superoxide production. However, structurally and mechanistically dissimilar inhibitors of protein methylation and coexpression of protein arginine methyltransferase 1 did not influence Nox activity. Rather, the effect of AMI-1 was rapidly reversible and could be demonstrated in an assay using chemically synthesized superoxide. We conclude that protein methylation does not regulate the activity of NADPH-oxidases and that AMI-1 is a potent antioxidant with a greater potency than 4,5-dihydroxy-1,3-benzenedisulfonic acid (Tiron) and 4 hydroxy-2,2,6,6-tetramethylpiperydine-1-oxyl (Tempol). PMID- 19903832 TI - 15-hydroxyeicosatetraenoic acid is a preferential peroxisome proliferator activated receptor beta/delta agonist. AB - Peroxisome proliferator-activated receptor (PPARs) modulate target gene expression in response to unsaturated fatty acid ligands, such as arachidonic acid (AA). Here, we report that the AA metabolite 15-hydroxyeicosatetraenoic acid (15-HETE) activates the ligand-dependent activation domain (AF2) of PPARbeta/delta in vivo, competes with synthetic agonists in a PPARbeta/delta ligand binding assay in vitro, and triggers the interaction of PPARbeta/delta with coactivator peptides. These agonistic effects were also seen with PPARalpha and PPARgamma, but to a significantly weaker extent. We further show that 15-HETE strongly induces the expression of the bona fide PPAR target gene Angptl4 in a PPARbeta/delta-dependent manner and, conversely, that inhibition of 15-HETE synthesis reduces PPARbeta/delta transcriptional activity. Consistent with its function as an agonistic ligand, 15-HETE triggers profound changes in chromatin associated PPARbeta/delta complexes in vivo, including the recruitment of the coactivator cAMP response element-binding protein binding protein. Both 15R-HETE and 15S-HETE are similarly potent at inducing PPARbeta/delta coactivator binding and transcriptional activation, indicating that 15-HETE enantiomers generated by different pathways function as PPARbeta/delta agonists. PMID- 19903833 TI - Prehypertensive preconditioning improves adult antihypertensive and cardioprotective treatment. AB - Transient prehypertensive treatment (TPT) causes prolonged antihypertensive and cardioprotective effects. Reduced angiotensin sensitivity has been attributed to those effects. We hypothesize that prehypertensive preconditioning by angiotensin II type 1 receptor (AT1R) blockade improves cardiovascular protection of late onset AT1R blockade in a model of spontaneous hypertensive heart-failure rats (SHHF/Mcc-fa(cp)). TPT (4-8 weeks of age) consisted of AT1R blockade (5 mg/kg candesartan) or vehicle. Candesartan-pretreated SHHF (5 mg/kg/day candesartan; weeks 4-8) received during adulthood (20-28 weeks of age) either candesartan at a dose of 1.5 or 5 mg/kg/day or vehicle. Vehicle-pretreated SHHF received either candesartan (5 mg/kg/day) or vehicle during adulthood. Blood pressure telemetry and longitudinal echocardiography were performed between weeks 20 and 28. Final examination included cardiac and vascular morphometry and measurement of the AT1R signaling and receptor internalization (ATRAP). Combined juvenile and adult AT1R blockade caused lower mean arterial pressure (MAP) than adult AT1R blockade alone (84 +/- 5 versus 97 +/- 5 mm Hg; P < 0.05). Cardiac and vascular hypertrophy was lower. Juvenile treatments were associated with a reduced cardiovascular AT1R expression and enhanced ATRAP expression. Combined juvenile and reduced adult AT1R blockade resulted in MAP similar to that with adult AT1R blockade alone (92 +/- 3 versus 97 +/- 5 mm Hg). We conclude that prehypertensive preconditioning improves adult treatment effects in SHHF. Those effects correlate with reduced cardiovascular AT1R expression and enhanced receptor internalization, suggesting reduced angiotensin sensitivity in pretreated SHHF. Moreover, preconditioning allows a reduction of adult AT1R blockade without loss of protection. Therefore, prehypertensive preconditioning may offer a tool to improve treatment efficacy in humans. PMID- 19903834 TI - Pharmacologic antagonism of the oral aversive taste-directed response to capsaicin in a mouse brief access taste aversion assay. AB - Chemosensory signaling by the tongue is a primary determinant of ingestive behavior and is mediated by specific interactions between tastant molecules and G protein-coupled and ion channel receptors. The functional relationship between tastant and receptor should be amenable to pharmacologic methods and manipulation. We have performed a pharmacologic characterization of the taste directed licking of mice presented with solutions of capsaicin and other transient receptor potential vanilloid-1 (TRPV1) agonists using a brief access taste aversion assay. Dose-response functions for lick-rate suppression were established for capsaicin (EC(50) = 0.5 microM), piperine (EC(50) = 2 muM), and resiniferatoxin (EC(50) = 0.02 microM). Little or no effect on lick rate was observed in response to the full TRPV1 agonist olvanil. Capsaicin lick rates of wild-type and transient receptor potential melastatin-5 (TRPM5) knockout mice were equivalent, indicating that TRPM5, a critical component of aversive signaling for many bitter tastants, did not contribute to the capsaicin taste response. The selective TRPV1 antagonists N-(4-tertiarybutylphenyl)-4-(3 chloropyridin-2-yl)tetrahydropyrazine-1(2H)-carbox-amide (10 microM) and (E)-3-(4 t-butylphenyl)-N-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)acrylamide (AMG9810) (10 microM) effectively blocked capsaicin- and piperine-mediated lick suppression. However, (E)-3-(4-chlorophenyl)-N-(3-methoxyphenyl)-N-phenylprop-2-enamide (SB 366791) and capsazepine, also TRPV1 antagonists, were without effect at test concentrations of up to 30 and 100 microM, respectively. Our results demonstrate that TRPV1-mediated oral aversiveness presents a pharmacologic profile differing from what has been reported previously for TRPV1 pain signaling and, furthermore, that aversive tastes can be evaluated and controlled pharmacologically. PMID- 19903839 TI - Curcumin potentiates the antitumor effects of Bacillus Calmette-Guerin against bladder cancer through the downregulation of NF-kappaB and upregulation of TRAIL receptors. AB - Although Bacillus Calmette-Guerin (BCG) intravesical therapy is a standard treatment for bladder cancer, eventual failure of response is a major problem. Treatments that can augment BCG therapy are urgently needed. We investigated whether curcumin, a component of Curcuma longa (also called turmeric), has potential to improve the current therapy using in vitro and in vivo MBT-2 murine tumor models. We found that curcumin potentiated BCG-induced apoptosis of human bladder cancer cells. BCG stimulated the release of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) from peripheral mononuclear neutrophils in a dose- and time-dependent manner, whereas curcumin enhanced the upregulation of TRAIL receptors. Electrophoretic mobility shift assay revealed that curcumin also suppressed the BCG-induced activation of the cell survival transcription factor NF-kappaB. In a syngeneic bladder cancer model, curcumin alone reduced the bladder tumor volume, but a significantly greater reduction was observed when BCG and curcumin were used in combination (P < 0.0001 versus control; P < 0.003 versus BCG alone). This was accompanied by a significant decrease in the proliferation marker Ki-67 (P < 0.01 versus control; P < 0.01 versus BCG alone) and microvessel density (CD31; P < 0.01 versus control; P < 0.01 versus BCG alone), decreased NF-kappaB in tumor tissue compared with the control, induced apoptosis, and decreased cyclin D1, vascular endothelial growth factor, cyclooxygenase-2, c-myc, and Bcl-2 expression in the tumor tissue. Upregulation of TRAIL receptor by the combination was also observed in tumor tissues. Overall, our results suggest that curcumin potentiates the antitumor effect of BCG through the inhibition of NF-kappaB and induction of TRAIL receptors in bladder cancer cells. PMID- 19903840 TI - Recognition and killing of brain tumor stem-like initiating cells by CD8+ cytolytic T cells. AB - Solid tumors contain a subset of stem-like cells that are resistant to the cytotoxic effects of chemotherapy/radiotherapy, but their susceptibility to cytolytic T lymphocyte (CTL) effector mechanisms has not been well characterized. Using a panel of early-passage human brain tumor stem/initiating cell (BTSC) lines derived from high-grade gliomas, we show that BTSCs are subject to immunologic recognition and elimination by CD8(+) CTLs. Compared with serum differentiated CD133(low) tumor cells and established glioma cell lines, BTSCs are equivalent with respect to expression levels of HLA class I and ICAM-1, similar in their ability to trigger degranulation and cytokine synthesis by antigen-specific CTLs, and equally susceptible to perforin-dependent CTL-mediated cytolysis. BTSCs are also competent in the processing and presentation of antigens as evidenced by the killing of these cells by CTL when antigen is endogenously expressed. Moreover, we show that CTLs can eliminate all BTSCs with tumor-initiating activity in an antigen-specific manner in vivo. Current models predict that curative therapies for many cancers will require the elimination of the stem/initiating population, and these studies lay the foundation for developing immunotherapeutic approaches to eradicate this tumor population. PMID- 19903841 TI - MiR-15a and MiR-16 control Bmi-1 expression in ovarian cancer. AB - Oncogenic activation of Bmi-1 is found in a wide variety of epithelial malignancies including ovarian cancer, yet a specific mechanism for overexpression of Bmi-1 has not been determined. Thus, realizing the immense pathologic significance of Bmi-1 in cancer, we wanted to investigate if microRNA (miRNA) aberrations played a role in the regulation of Bmi-1 in ovarian cancer. In this report, we identify two miRNAs, miR-15a and miR-16, that are underexpressed in ovarian cell lines and in primary ovarian tissues. We show that these miRNAs directly target the Bmi-1 3' untranslated region and significantly correlate with Bmi-1 protein levels in ovarian cancer patients and cell lines. Furthermore, Bmi-1 protein levels are downregulated in response to miR-15a or miR 16 expression and lead to significant reduction in ovarian cancer cell proliferation and clonal growth. These findings suggest the development of therapeutic strategies by restoring miR-15a and miR-16 expression in ovarian cancer and in other cancers that involve upregulation of Bmi-1. PMID- 19903842 TI - Dynamic gene expression analysis links melanocyte growth arrest with nevogenesis. AB - Like all primary cells in vitro, normal human melanocytes exhibit a physiologic decay in proliferative potential as it transitions to a growth-arrested state. The underlying transcriptional program(s) that regulate this phenotypic change is largely unknown. To identify molecular determinants of this process, we performed a Bayesian-based dynamic gene expression analysis on primary melanocytes undergoing proliferative arrest. This analysis revealed several related clusters whose expression behavior correlated with the melanocyte growth kinetics; we designated these clusters the melanocyte growth arrest program (MGAP). These MGAP genes were preferentially represented in benign melanocytic nevi over melanomas and selectively mapped to the hepatocyte fibrosis pathway. This transcriptional relationship between melanocyte growth stasis, nevus biology, and fibrogenic signaling was further validated in vivo by the demonstration of strong pericellular collagen deposition within benign nevi but not melanomas. Taken together, our study provides a novel view of fibroplasia in both melanocyte biology and nevogenesis. PMID- 19903843 TI - Autophagy is activated by TGF-beta and potentiates TGF-beta-mediated growth inhibition in human hepatocellular carcinoma cells. AB - Transforming growth factor-beta (TGF-beta) is a multifunctional cytokine that regulates cell growth, differentiation, and apoptosis of various types of cells. Autophagy is emerging as a critical response of normal and cancer cells to environmental changes, but the relationship between TGF-beta signaling and autophagy has been poorly understood. Here, we showed that TGF-beta activates autophagy in human hepatocellular carcinoma cell lines. TGF-beta induced accumulation of autophagosomes and conversion of microtubule-associated protein 1 light chain 3 and enhanced the degradation rate of long-lived proteins. TGF-beta increased the mRNA expression levels of BECLIN1, ATG5, ATG7, and death-associated protein kinase (DAPK). Knockdown of Smad2/3, Smad4, or DAPK, or inhibition of c Jun NH(2)-terminal kinase, attenuated TGF-beta-induced autophagy, indicating the involvement of both Smad and non-Smad pathway(s). TGF-beta activated autophagy earlier than execution of apoptosis (6-12 versus 48 h), and reduction of autophagy genes by small interfering RNA attenuated TGF-beta-mediated growth inhibition and induction of proapoptotic genes Bim and Bmf, suggesting the contribution of autophagy pathway to the growth-inhibitory effect of TGF-beta. Additionally, TGF-beta also induced autophagy in some mammary carcinoma cells, including MDA-MB-231 cells. These findings show that TGF-beta signaling pathway activates autophagy in certain human cancer cells and that induction of autophagy is a novel aspect of biological functions of TGF-beta. PMID- 19903844 TI - De novo discovery of a gamma-secretase inhibitor response signature using a novel in vivo breast tumor model. AB - Notch pathway signaling plays a fundamental role in normal biological processes and is frequently deregulated in many cancers. Although several hypotheses regarding cancer subpopulations most likely to respond to therapies targeting the Notch pathway have been proposed, clinical utility of these predictive markers has not been shown. To understand the molecular basis of gamma-secretase inhibitor (GSI) sensitivity in breast cancer, we undertook an unbiased, de novo responder identification study using a novel genetically engineered in vivo breast cancer model. We show that tumors arising from this model are heterogeneous on the levels of gene expression, histopathology, growth rate, expression of Notch pathway markers, and response to GSI treatment. In addition, GSI treatment of this model was associated with inhibition of Hes1 and proliferation markers, indicating that GSI treatment inhibits Notch signaling. We then identified a pretreatment gene expression signature comprising 768 genes that is significantly associated with in vivo GSI efficacy across 99 tumor lines. Pathway analysis showed that the GSI responder signature is enriched for Notch pathway components and inflammation/immune-related genes. These data show the power of this novel in vivo model system for the discovery of biomarkers predictive of response to targeted therapies, and provide a basis for the identification of human breast cancers most likely to be sensitive to GSI treatment. PMID- 19903846 TI - Persistence of betapapillomavirus infections as a risk factor for actinic keratoses, precursor to cutaneous squamous cell carcinoma. AB - Human papillomaviruses from the beta genus (betaPV) are a possible cause of cutaneous squamous cell carcinoma (SCC). We assessed the extent to which betaPV infections persisted long-term in a subtropical Australian community and whether betaPV persistence is positively associated with actinic keratoses, precursor for SCC. Eyebrow hairs were collected from 171 participants of the community-based Nambour Skin Cancer Study in 1996 and 2003. Hair samples were tested for the presence of DNA from 25 different betaPV types and assessed in relation to actinic keratosis presence in 2007. In 1996, a total of 413 betaPV infections were found in 73% of participants, increasing to 490 infections among 85% in 2003. Of the total betaPV infections detected, 211 (30%) were found to persist. Age was significantly associated with betaPV persistence: those ages >60 years had 1.5-fold (95% confidence interval, 1.1-1.9) increased risk of type-specific viral persistence than those ages <40 years. After accounting for actinic keratoses at baseline, persistence of betaPV DNA resulted in a 1.4-fold (95% confidence interval, 1.0-1.9) increase in risk of having actinic keratoses on the face in 2007. In conclusion, persistent betaPV infections in this population were associated with an increased occurrence of actinic keratosis. Additional studies are needed to determine the possible association of betaPV persistence with SCC. PMID- 19903845 TI - Differential enhancement of breast cancer cell motility and metastasis by helical and kinase domain mutations of class IA phosphoinositide 3-kinase. AB - Class IA (p85/p110) phosphoinositide 3-kinases play a major role in regulating cell growth, survival, and motility. Activating mutations in the p110alpha isoform of the class IA catalytic subunit (PIK3CA) are commonly found in human cancers. These mutations lead to increased proliferation and transformation in cultured cells, but their effects on cell motility and tumor metastasis have not been evaluated. We used lentiviral-mediated gene transfer and knockdown to produce stable MDA-MB-231 cells in which the endogenous human p110alpha is replaced with either wild-type bovine p110alpha or the two most common activating p110alpha mutants, the helical domain mutant E545K and the kinase domain mutant H1047R. The phosphoinositide 3-kinase/Akt pathway was hyperactivated in cells expressing physiologic levels of helical or kinase domain mutants. Cells expressing either mutant showed increased motility in vitro, but only cells expressing the helical domain mutant showed increased directionality in a chemotaxis assay. In severe combined immunodeficient mice, xenograft tumors expressing either mutant showed increased rates of tumor growth compared with tumors expressing wild-type p110alpha. However, tumors expressing the p110alpha helical domain mutant showed a marked increase in both tumor cell intravasation into the blood and tumor cell extravasation into the lung after tail vein injection compared with tumors expressing wild-type p110alpha or the kinase domain mutant. Our observations suggest that, when compared with kinase domain mutations in a genetically identical background, expression of helical domain mutants of p110alpha produce a more severe metastatic phenotype. PMID- 19903847 TI - Breast cancer amplified sequence 2, a novel negative regulator of the p53 tumor suppressor. AB - Breast cancer amplified sequence 2 (BCAS2) was reported previously as a transcriptional coactivator of estrogen receptor. Here, we report that BCAS2 directly interacts with p53 to reduce p53 transcriptional activity by mildly but consistently decreasing p53 protein in the absence of DNA damage. However, in the presence of DNA damage, BCAS2 prominently reduces p53 protein and provides protection against chemotherapeutic agent such as doxorubicin. Deprivation of BCAS2 induces apoptosis in p53 wild-type cells but causes G(2)-M arrest in p53 null or p53 mutant cells. There are at least two apoptosis mechanisms induced by silencing BCAS2 in wild-type p53-containing cells. Firstly, it increases p53 retention in nucleus that triggers the expression of apoptosis-related genes. Secondly, it increases p53 transcriptional activity by raising p53 phosphorylation at Ser(46) and decreases p53 protein degradation by reducing p53 phosphorylation at Ser(315). We show for the first time that BCAS2, a small nuclear protein (26 kDa), is a novel negative regulator of p53 and hence a potential molecular target for cancer therapy. PMID- 19903848 TI - Estradiol alters cell growth in nonmalignant colonocytes and reduces the formation of preneoplastic lesions in the colon. AB - Numerous clinical and animal studies show that hormone replacement therapy reduces the risk of colon tumor formation. However, the majority of experiments have shown that estradiol (E(2)) does not inhibit the growth of malignantly transformed colon epithelia. As such, the presented studies focused on evaluating the effects of E(2) in noncancerous colonocytes. E(2) treatments (0-10 nmol/L) reduced cell growth and increased apoptotic activity in young adult mouse colonocytes (YAMC), a nonmalignant cell line, in a dose-responsive manner. These effects were lost in the YAMC-Ras cells, an isogenic cell line with a single malignant transformation. Cotreatment with an estrogen receptor (ER) antagonist inhibited the physiologic effects of E(2) in YAMC cells, suggesting that the response is ER mediated. To further study the effect of E(2) on colonic epithelia, we evaluated the development of preneoplastic lesions in ovariectomized wild-type (WT) and ERbeta knockout (ERbetaKO) mice treated with either vehicle or E(2). WT E(2)-treated animals exhibited significantly fewer aberrant crypt foci and increased apoptotic activity in colonic epithelia when compared with WT control mice or ERbetaKO animals receiving either treatment. For the first time, we showed that E(2) alters the growth of nontransformed colonocytes in vitro and that, through an ERbeta-mediated mechanism, E(2) influences the physiology of noncancerous colonocytes, resulting in fewer preneoplastic lesions. Collectively, these data show that the protective actions of E(2) occur primarily during the initiation/promotion stages of disease development and identify the hormone as an important chemoprotective agent. PMID- 19903849 TI - Tangeretin sensitizes cisplatin-resistant human ovarian cancer cells through downregulation of phosphoinositide 3-kinase/Akt signaling pathway. AB - Combination of innocuous dietary components with anticancer drugs is an emerging new strategy for cancer chemotherapy to increase antitumor responses. Tangeretin is a citrus flavonoid known to inhibit cancer cell proliferation. Here, we show an enhanced response of A2780/CP70 and 2008/C13 cisplatin-resistant human ovarian cancer cells to various combination treatments of cisplatin and tangeretin. Pretreatment of cells with tangeretin before cisplatin treatment synergistically inhibited cancer cell proliferation. This combination was effective in activating apoptosis via caspase cascade as well as arresting cell cycle at G(2)-M phase. Moreover, phospho-Akt and its downstream substrates, e.g., NF-kappaB, phospho-GSK 3beta, and phospho-BAD, were downregulated upon tangeretin-cisplatin treatment. The tangeretin-cisplatin-induced apoptosis in A2780/CP70 cells was increased by phosphoinositide-3 kinase (PI3K) inhibition and siRNA-mediated Akt silencing, but reduced by overexpression of constitutively activated Akt and GSK-3beta inhibition. The overall results indicated that tangeretin exposure preconditions cisplatin-resistant human ovarian cancer cells for a conventional response to low dose cisplatin-induced cell death occurring through downregulation of PI3K/Akt signaling pathway. Thus, effectiveness of tangeretin combinations, as a promising modality in the treatment of resistant cancers, warrants systematic clinical studies. PMID- 19903850 TI - Detection of treatment-induced changes in signaling pathways in gastrointestinal stromal tumors using transcriptomic data. AB - Cell signaling plays a central role in the etiology of cancer. Numerous therapeutics in use or under development target signaling proteins; however, off target effects often limit assignment of positive clinical response to the intended target. As direct measurements of signaling protein activity are not generally feasible during treatment, there is a need for more powerful methods to determine if therapeutics inhibit their targets and when off-target effects occur. We have used the Bayesian Decomposition algorithm and data on transcriptional regulation to create a novel methodology, Differential Expression for Signaling Determination (DESIDE), for inferring signaling activity from microarray measurements. We applied DESIDE to deduce signaling activity in gastrointestinal stromal tumor cell lines treated with the targeted therapeutic imatinib mesylate (Gleevec). We detected the expected reduced activity in the KIT pathway, as well as unexpected changes in the p53 pathway. Pursuing these findings, we have determined that imatinib-induced DNA damage is responsible for the increased activity of p53, identifying a novel off-target activity for this drug. We then used DESIDE on data from resected, post-imatinib treatment tumor samples and identified a pattern in these tumors similar to that at late time points in the cell lines, and this pattern correlated with initial clinical response. The pattern showed increased activity of ETS domain-containing protein Elk-1 and signal transducers and activators of transcription 3 transcription factors, which are associated with the growth of side population cells. DESIDE infers the global reprogramming of signaling networks during treatment, permitting treatment modification that leverages ongoing drug development efforts, which is crucial for personalized medicine. PMID- 19903851 TI - Calcineurin inhibitors activate the proto-oncogene Ras and promote protumorigenic signals in renal cancer cells. AB - The development of cancer is a major problem in immunosuppressed patients, particularly after solid organ transplantation. We have recently shown that calcineurin inhibitors (CNI) used to treat transplant patients may play a critical role in the rapid progression of renal cancer. To examine the intracellular signaling events for CNI-mediated direct tumorigenic pathway(s), we studied the effect of CNI on the activation of proto-oncogenic Ras in human normal renal epithelial cells (REC) and renal cancer cells (786-0 and Caki-1). We found that CNI treatment significantly increased the level of activated GTP-bound form of Ras in these cells. In addition, CNI induced the association of Ras with one of its effector molecules, Raf, but not with Rho and phosphatidylinositol 3 kinase; CNI treatment also promoted the phosphorylation of the Raf kinase inhibitory protein and the downregulation of carabin, all of which may lead to the activation of the Ras-Raf pathway. Blockade of this pathway through either pharmacologic inhibitors or gene-specific small interfering RNA significantly inhibited CNI-mediated augmented proliferation of renal cancer cells. Finally, it was observed that CNI treatment increased the growth of human renal tumors in vivo, and the Ras-Raf pathway is significantly activated in the tumor tissues of CNI-treated mice. Together, targeting the Ras-Raf pathway may prevent the development/progression of renal cancer in CNI-treated patients. PMID- 19903852 TI - Optimized peptide vaccines eliciting extensive CD8 T-cell responses with therapeutic antitumor effects. AB - A major challenge for developing effective therapeutic vaccines against cancer is overcoming immunologic tolerance to tumor-associated antigens that are expressed on both malignant cells and normal tissues. Herein, we describe a novel vaccination approach, TriVax, that uses synthetic peptides representing CD8 T cell epitopes, Toll-like receptor agonists that function as potent immunologic adjuvants and costimulatory anti-CD40 antibodies to generate large numbers of high-avidity antigen-reactive T cells capable of recognizing and killing tumor cells. Our results show that TriVax induced huge numbers of long-lasting antigen specific CD8 T cells that displayed significant antitumor effects in vivo. The administration of a TriVax formulation containing a CD8 T-cell epitope derived from a melanosomal antigen (Trp2(180-188)) elicited antigen-specific CD8 T cells that induced systemic autoimmunity (vitiligo). More important, TriVax immunization was effective in eliciting potent protective antitumor immunity as well as remarkable therapeutic effects against established B16 melanoma. This therapeutic effect was mediated by CD8 T cells via perforin-mediated lysis and required the participation of type-I IFN but not IFNgamma. These results suggest that similar strategies would be applicable for the design of effective vaccination for conducting clinical studies in cancer patients. PMID- 19903853 TI - Improved expression and reactivity of transduced tumor-specific TCRs in human lymphocytes by specific silencing of endogenous TCR. AB - Adoptive T-cell therapy using lymphocytes genetically engineered to express tumor antigen-specific TCRs is an attractive strategy for treating patients with malignancies. However, there are potential drawbacks to this strategy: mispairing of the introduced TCR alpha/beta chains with the endogenous TCR subunits and competition of CD3 molecules between the introduced and endogenous TCRs can impair cell surface expression of the transduced TCR, resulting in insufficient function and potential generation of autoreactive T cells. In addition, the risk of tumor development following the infusion of cells with aberrant vector insertion sites increases with the vector copy number in the transduced cells. In this study, we developed retroviral vectors encoding both small interfering RNA constructs that specifically down-regulate endogenous TCR and a codon-optimized, small interfering RNA-resistant TCR specific for the human tumor antigens MAGE-A4 or WT1. At low copy numbers of the integrated vector, the transduced human lymphocytes exhibited high surface expression of the introduced tumor-specific TCR and reduced expression of endogenous TCRs. In consequence, the vector transduced lymphocytes showed enhanced cytotoxic activity against antigen expressing tumor cells. Therefore, our novel TCR gene therapy may open a new gate for effective immunotherapy in cancer patients. PMID- 19903854 TI - Astrocyte elevated gene-1: far more than just a gene regulated in astrocytes. AB - Since its original cloning by subtraction hybridization in 2002, it is now evident that Astrocyte elevated gene-1 (AEG-1) is a key contributor to the carcinogenic process in diverse organs. AEG-1 protein expression is elevated in advanced stages of many cancers, which correlates with poor survival. In specific cancers, such as breast and liver cancer, the AEG-1 gene itself is amplified, further supporting a seminal role in tumorigenesis. Overexpression and inhibition studies both in in vitro and in in vivo models reveal the importance of AEG-1 in regulating multiple physiologically and pathologically relevant processes including proliferation, invasion, metastasis, and gene expression. AEG-1 is a single-pass transmembrane protein with multiple nuclear localization signals and no known domains or motifs. Although pertinent roles of AEG-1 in the carcinogenic process are established, its potential function (promotion of metastasis only versus functioning as a bona fide oncogene) as well as localization (cell surface versus nucleus) remain areas requiring further clarification. The present review critically evaluates what is currently known about AEG-1 and provides new perspectives relative to this intriguing molecule that may provide a rational target for intervening in the cancer phenotype. PMID- 19903855 TI - The fibroblast growth factor receptor inhibitor PD173074 blocks small cell lung cancer growth in vitro and in vivo. AB - Lung cancer is the commonest cancer killer. Small cell lung cancer (SCLC) is initially chemosensitive, but rapidly relapses in a chemoresistant form with an overall survival of <5%. Consequently, novel therapies are urgently required and will likely arise from an improved understanding of the disease biology. Our previous work showed that fibroblast growth factor-2 induces proliferation and chemoresistance in SCLC cells. Here, we show that the selective fibroblast growth factor receptor (FGFR) inhibitor PD173074 blocks H-510 and H-69 SCLC proliferation and clonogenic growth in a dose-dependent fashion and prevents FGF 2-induced chemoresistance. These effects correlate with the inhibition of both FGFR1 and FGFR2 transphosphorylation. We then determined the efficacy of daily oral administration of PD173074 for 28 days in two human SCLC models. In the H 510 xenograft, tumor growth was impaired similar to that seen with single-agent cisplatin administration, increasing median survival compared with control sham treated animals. Crucially, the effect of cisplatin was significantly potentiated by coadministration of PD173074. More dramatically, in H-69 xenografts, PD173074 induced complete responses lasting >6 months in 50% of mice. These effects were not a consequence of disrupted tumor vasculature but instead correlated with increased apoptosis (caspase 3 and cytokeratin 18 cleavage) in excised tumors. Moreover, in vivo imaging with 3'-deoxy-3'-[(18)F]fluorothymidine-positron emission tomography ([(18)F]FLT-PET) showed decreased intratumoral proliferation in live animals treated with the compound at 7 to 14 days. Our results suggest that clinical trials of FGFR inhibitors should be undertaken in patients with SCLC and that [(18)F]FLT-PET imaging could provide early in vivo evidence of response. PMID- 19903856 TI - Loss of collapsin response mediator Protein1, as detected by iTRAQ analysis, promotes invasion of human gliomas expressing mutant EGFRvIII. AB - Glioblastoma multiforme (GBM) is the most common and lethal primary human brain tumor. GBMs are characterized by a variety of genetic alterations, among which oncogenic mutations of epidermal growth factor receptor (EGFRvIII) is most common. GBMs harboring EGFRvIII have increased proliferation and invasive characteristics versus those expressing wild-type (wt) EGFR. To identify the molecular basis of this increased tumorgenic phenotype, we used iTRAQ-labeling differential proteomic analysis. Among several differentially expressed proteins, we selected CRMP1, a protein implicated in cellular invasion that was markedly decreased in GBMs expressing EGFRvIII, for further study. The differential expression of CRMP1 was confirmed in a panel of human GBM cell lines and operative specimens that express wtEGFR or mutant EGFRvIII by quantitative real time PCR, Western blot, and immunohistochemical analysis. In human GBM samples, decreased expression of CRMP1 correlated with EGFRvIII positivity. Knockdown of CRMP1 by siRNA resulted in increased invasion of wtEGFR expressing human GBM cells (U87 and U373) to those found in isogenic GBM cells. Exogenous expression of EGFRvIII in these wtEGFR-expressing GBM cells promoted their ability to invade and was accompanied by decreased expression of CRMP1. Rescuing CRMP1 expression decreased invasion of the EGFRvIII-expressing GBM cells by tilting the balance between Rac and Rho. Collectively, these results show that the loss of CRMP1 contribute to the increased invasive phenotype of human GBMs expressing mutant EGFRvIII. PMID- 19903857 TI - Targeted disruption of the S1P2 sphingosine 1-phosphate receptor gene leads to diffuse large B-cell lymphoma formation. AB - S1P(2) sphingosine 1-phosphate receptor signaling can regulate proliferation, survival, morphology, and migration in many cell types in vitro. Here, we report that S1P(2)(-/-) mice develop clonal B-cell lymphomas with age, such that approximately half of the animals display this neoplasm by 1.5 to 2 years of age. Histologic, immunophenotypic, and molecular analyses revealed a uniform tumor phenotype with features of germinal center (GC)-derived diffuse large B-cell lymphoma (DLBCL). Tumor formation was preceded by increases in GC B cells and CD69(+) T cells, as well as an increased formation of spontaneous GCs, suggesting that S1P(2) loss may promote lymphomagenesis in part by disrupting GC B-cells homeostasis. With the sole exception of rare lung tumors, the effect of S1P(2) gene disruption is remarkably restricted to DLBCL. In humans, 28 of 106 (26%) DLBCL samples were found to harbor multiple somatic mutations in the 5' sequences of the S1P(2) gene. Mutations displayed features resembling those generated by the IgV-associated somatic hypermutation mechanism, but were not detected at significant levels in normal GC B cells, indicating a tumor-associated aberrant function. Collectively, our data suggest that S1P(2) signaling may play a critical role in suppressing DLBCL formation in vivo. The high incidence of DLBCL in S1P(2)(-/-) mice, its onset at old age, and the relative lack of other neoplasms identify these mice as a novel, and potentially valuable, model for this highly prevalent and aggressive human malignancy. PMID- 19903858 TI - De-N-acetyl GM3 promotes melanoma cell migration and invasion through urokinase plasminogen activator receptor signaling-dependent MMP-2 activation. AB - We have recently discovered that de-N-acetyl GM3 [NeuNH(2)LacCer, d-GM3], a derivative of ganglioside GM3, is specifically expressed in metastatic tumor cells and that its expression correlates with an enhanced metastatic phenotype. Although the classic N-acetylated form of GM3 (NeuAcLacCer, c-GM3) is found in both normal and tumor cells, metastatic tumor cells (but not other cells) predominantly express d-GM3 (82-95% of total GM3). d-GM3 expression is mainly found in metastatic melanomas, but not in benign nevi or the majority of primary melanomas. Using metastatic (d-GM3-positive) and poorly invasive (d-GM3-negative) human melanoma cell lines, we found that d-GM3 stimulates cell migration and invasion by increasing the expression and activation of urokinase-like plasminogen activator (uPA). Further studies showed that d-GM3 activates matrix metalloproteinase-2 (MMP-2), but not MMP-9, when uPA receptor signaling is activated. These results implicate d-GM3 as a specific marker for metastatic melanoma and a novel therapeutic target for neoplastic diseases. PMID- 19903859 TI - An approach for addressing the multiple testing problem in social policy impact evaluations. AB - In social policy evaluations, the multiple testing problem occurs due to the many hypothesis tests that are typically conducted across multiple outcomes and subgroups, which can lead to spurious impact findings. This article discusses a framework for addressing this problem that balances Types I and II errors. The framework involves specifying confirmatory and exploratory analyses in study protocols, delineating confirmatory outcome domains, conducting t tests on composite domain outcomes, and applying multiplicity corrections to composites across domains to obtain summative impact evidence. The article presents statistical background and discusses multiplicity issues for subgroup analyses, designs with multiple treatments, and reporting. PMID- 19903860 TI - Association between exposure to topical tacrolimus or pimecrolimus and cancers. AB - BACKGROUND: The Food and Drug Administration has issued a public health advisory regarding cancer risk from topical calcineurin inhibitors. OBJECTIVE: To compare the rates of cancer among patients with common dermatologic conditions who were exposed or not exposed to topical calcineurin inhibitors. METHODS: A retrospective cohort observational study used data from an integrated healthcare delivery system on 953,064 subjects with diagnoses of atopic dermatitis or eczema between 2001 and December 2004. The main endpoint was initial cancer diagnosis. Chart review was performed to confirm cancer diagnosis in the subjects exposed to topical calcineurin inhibitors when any particular cancer rate was at least 3 times higher than that in unexposed subjects. Data were analyzed using the Cox proportional hazards model. RESULTS: Age- and sex-adjusted hazard ratios for all cancers were 0.93 (95% CI 0.81 to 1.07; p = 0.306) for tacrolimus-exposed versus unexposed subjects and 1.15 (95% CI 0.99 to 1.31; p = 0.054) for pimecrolimus exposed versus -unexposed subjects. T-cell lymphoma was the only cancer associated with a significantly increased risk among subjects exposed to tacrolimus (HR = 5.04, 95% CI 2.39 to 10.63; p < 0.001) or pimecrolimus (HR = 3.76, 95% CI 1.71 to 8.28; p = 0.010). Subsequent chart review of subjects in the exposed group with T-cell lymphoma found that 4 of 16 had skin lesions that were suspected to be the early lesions of T-cell lymphoma prior to exposure to tacrolimus or pimecrolimus. After these 4 cases were excluded, the age and sex hazard ratio for T-cell lymphoma was 5.44 (95% CI 2.51 to 11.79; p < 0.001) for tacrolimus and 2.32 (95% CI 0.89 to 6.07; p = 0.086) for pimecrolimus. There was no statistically significantly increased risk for other subgroups of cancer, including melanoma. CONCLUSIONS: Exposure to topical tacrolimus or pimecrolimus was not associated with an increase in the overall cancer rate. Use of topical tacrolimus may be associated with an increased risk of T-cell lymphoma. PMID- 19903861 TI - Colchicine for the primary and secondary prevention of pericarditis: an update. AB - OBJECTIVE: To review the efficacy and safety of colchicine as primary and secondary prophylaxis for pericarditis. DATA SOURCES: We searched MEDLINE, EMBASE, PubMed, BIOSIS Previews, International Pharmaceutical Abstracts, Web of Science, and CENTRAL for controlled studies from database inception date to July 2009. Search terms included colchicine, pericarditis, and postpericardiotomy syndrome (PPS). STUDY SELECTION AND DATA EXTRACTION: Prospective, randomized, controlled trials investigating the use of colchicine in preventing pericarditis were included. Data extracted included design, inclusion criteria, demographics, interventions, background therapy, and pericarditis-related clinical outcomes. DATA SYNTHESIS: Data were synthesized qualitatively, given variable study designs. Three trials were identified. A single trial examining primary prevention evaluated the use of colchicine versus placebo for preventing PPS in patients undergoing cardiopulmonary bypass grafting. No significant reduction in PPS was found. Two studies examined secondary prevention of pericarditis, comparing colchicine plus aspirin versus aspirin alone. One study examined using these comparators to treat a first episode of pericarditis. After 3 months, there was a significant reduction in recurrent pericarditis with colchicine plus aspirin (11.7% vs 33%; p = 0.009). Another study examined this same regimen in recurrent pericarditis, finding a significant reduction in recurrence after 6 months (21% vs 45%; p = 0.02). CONCLUSIONS: Despite limitations in study designs, current evidence suggests a role for colchicine in the secondary prophylaxis for recurrent pericarditis. The evidence for use of colchicine as primary prophylaxis in PPS is indeterminate; therefore, colchicine cannot be recommended routinely. While colchicine should be recommended for the prevention of recurrent pericarditis, questions regarding the optimal regimen and long-term safety profile need to be further elucidated. PMID- 19903862 TI - Influence of glucosamine on glomerular mesangial cell turnover: implications for hyperglycemia and hexosamine pathway flux. AB - Cells exposed to high glucose may undergo hypertrophy, proliferation, and apoptosis, but the role of hexosamine flux in mediating these effects has not been fully elucidated. Accordingly, we studied the effects of glucose and glucosamine on rat glomerular mesangial cells (MC) turnover. Compared with physiological glucose (5.6 mM), treatment with high glucose (25 mM) for 24 h stimulated MC proliferation, an effect that was mimicked by exposure to low concentrations of glucosamine (0.05 mM). The percentage of cells in G(0)/G(1) phase of the cell cycle was reduced with a concomitant increase of the number of cells in G(2)/M phase. Proliferating cell nuclear antigen, phosphorylated mammalian target of rapamycin [phospho-mTOR (Ser(2448))], and total regulatory associated protein of mTOR were increased by high glucose and glucosamine treatment. Inhibition of glutamine:fructose-6-phosphate amidotransferase (GFAT), the rate-limiting enzyme for hexosamine flux, with 6-diazo-5-oxonorleucine (10 muM) and of mTOR with rapamycin both attenuated glucose-mediated MC proliferation. Higher glucosamine concentrations (0.25-10 mM) caused MC apoptosis after 48 h, and, in addition, GFAT overexpression also increased MC apoptosis (TdT-dUTP nick end-labeling-positive cells: 3.8 +/- 0.3 vs. 1.1 +/- 0.2% for empty vector; P < 0.05). Hence, hexosamine flux is an important determinant of MC proliferation and apoptosis. The proliferative response to high glucose and hexosamine flux is rapamycin-sensitive, suggesting that this effect is associated with signaling through rapamycin-sensitive mTOR complex 1 (mTORC1). PMID- 19903863 TI - Parenteral and enteral metabolism of anaplerotic triheptanoin in normal rats. II. Effects on lipolysis, glucose production, and liver acyl-CoA profile. AB - The anaplerotic odd-medium-chain triglyceride triheptanoin is used in clinical trials for the chronic dietary treatment of patients with long-chain fatty acid oxidation disorders. We previously showed (Kinman RP, Kasumov T, Jobbins KA, Thomas KR, Adams JE, Brunengraber LN, Kutz G, Brewer WU, Roe CR, Brunengraber H. Am J Physiol Endocrinol Metab 291: E860-E866, 2006) that the intravenous infusion of triheptanoin increases lipolysis traced by the turnover of glycerol. In this study, we tested whether lipolysis induced by triheptanoin infusion is accompanied by the potentially harmful release of long-chain fatty acids. Rats were infused with heptanoate +/- glycerol or triheptanoin. Intravenous infusion of triheptanoin at 40% of caloric requirement markedly increased glycerol endogenous R(a) but not oleate endogenous R(a). Thus, the activation of lipolysis was balanced by fatty acid reesterification in the same cells. The liver acyl-CoA profile showed the accumulation of intermediates of heptanoate beta-oxidation and C(5)-ketogenesis and a decrease in free CoA but no evidence of metabolic perturbation of liver metabolism such as propionyl overload. Our data suggest that triheptanoin, administered either intravenously or intraduodenally, could be used for intensive care and nutritional support of metabolically decompensated long-chain fatty acid oxidation disorders. PMID- 19903864 TI - Endothelial function in subjects with isolated low HDL cholesterol: role of nitric oxide and circulating progenitor cells. AB - Epidemiologic studies have shown that a low level of high-density lipoprotein (HDL) cholesterol is a risk factor for cardiovascular diseases. The purpose of this study was to determine the contribution of isolated low HDL cholesterol to endothelial function. Thirty-nine subjects with low HDL cholesterol who had no other cardiovascular risk factors were selected from the 5,417 participants from our population. We evaluated flow-mediated vasodilation (FMD) before and after 4 wk of treatment with the HMG-CoA reductase inhibitor pravastatin in 29 of the 39 subjects with isolated low HDL cholesterol. FMD was lower in the low-HDL cholesterol group (n = 29) than in the control group (n = 29), whereas NTG induced vasodilation was similar in the two groups. Pravastatin increased HDL cholesterol, urinary excretion of nitrite/nitrate, circulating levels of progenitor cells, and cell migration response to vascular endothelial growth factor in 15 subjects with low HDL cholesterol but not in 14 placebo control subjects. FMD increased in the pravastatin treatment group but not in the control group. NTG-induced vasodilation was similar before and after 4 wk of treatment in the two groups. Multiple regression analysis revealed that changes in HDL cholesterol, the number of progenitor cells, and migration of progenitor cells were independent predictors of augmentation of FMD with pravastatin. These findings suggest that low HDL cholesterol is an independent risk factor for endothelial dysfunction and that pravastatin improves endothelial function in individuals with isolated low HDL cholesterol through, at least in part, an increase in circulating progenitor cells. PMID- 19903865 TI - Transcriptional coactivator p300 regulates glucose-induced gene expression in endothelial cells. AB - Sustained hyperglycemia in diabetes causes alteration of a large number of transcription factors and mRNA transcripts, leading to tissue damage. We investigated whether p300, a transcriptional coactivator with histone acetyl transferase activity, regulates glucose-induced activation of transcription factors and subsequent upregulation of vasoactive factors and extracellular matrix (ECM) proteins in human umbilical vein endothelial cells (HUVECs). HUVECs were incubated in varied glucose concentrations and were studied after p300 small interfering RNA (siRNA) transfection, p300 overexpression, or incubation with the p300 inhibitor curcumin. Histone H2AX phosphorylation and lysine acetylation were examined for oxidative DNA damage and p300 activation. Screening for transcription factors was performed with the Luminex system. Alterations of selected transcription factors were validated. mRNA expression of p300, endothelin-1 (ET-1), vascular endothelial growth factor (VEGF), and fibronectin (FN) and its splice variant EDB(+)FN and FN protein production were analyzed. HUVECs in 25 mmol/l glucose showed increased p300 production accompanied by increased binding of p300 to ET-1 and FN promoters, augmented histone acetylation, H2AX phosphorylation, activation of multiple transcription factors, and increased mRNA expression of vasoactive factors and ECM proteins. p300 overexpression showed a glucose-like effect on the mRNA expression of ET-1, VEGF, and FN. Furthermore, siRNA-mediated p300 blockade or chemical inhibitor of p300 prevented such glucose-induced changes. Similar mRNA upregulation was also seen in the organ culture of vascular tissues, which was prevented by p300 siRNA transfection. Data from these studies suggest that glucose-induced p300 upregulation is an important upstream epigenetic mechanism regulating gene expression of vasoactive factors and ECM proteins in endothelial cells and is a potential therapeutic target for diabetic complications. PMID- 19903866 TI - Contraction intensity and feeding affect collagen and myofibrillar protein synthesis rates differently in human skeletal muscle. AB - Exercise stimulates muscle protein fractional synthesis rate (FSR), but the importance of contractile intensity and whether it interplays with feeding is not understood. This was investigated following two distinct resistance exercise (RE) contraction intensities using an intrasubject design in the fasted (n = 10) and fed (n = 10) states. RE consisted of 10 sets of knee extensions. One leg worked against light load (LL) at 16% of one-repetition maximum (1RM), the other leg against heavy load (HL) at 70% 1RM, with intensities equalized for total lifted load. Males were infused with [(13)C]leucine, and vastus lateralis biopsies were obtained bilaterally at rest as well as 0.5, 3, and 5.5 h after RE. Western blots were run on muscle lysates and phosphospecific antibodies used to detect phosphorylation status of targets involved in regulation of FSR. The intramuscular collagen FSR was evenly increased following LL- and HL-RE and was not affected by feeding. Myofibrillar FSR was unaffected by LL-RE, whereas HL-RE resulted in a delayed improvement (0.14 +/- 0.02%/h, P < 0.05). Myofibrillar FSR was increased at rest by feeding (P < 0.05) and remained elevated late in the postexercise period compared with the fasting condition. The Rp-s6k-4E-binding protein-1 (BP1) and the mitogen-activated protein kinase (MAPk) pathways were activated by the HL intensity and were suggested to be responsible for regulating myofibrillar FSR in response to adequate contractile activity. Feeding predominantly affected Rp-s6k and eukaryotic elongation factor 2 phosphorylations in correspondence with the observed changes in myofibrillar FSR, whereas 4E-BP1 remained to respond only to the HL contraction intensity. Thus the study design allows us to conclude that the MAPk- and mammalian target of rapamycin-dependent signaling responds to contractile activity, whereas elongation mainly was found to respond to feeding. Furthermore, although functionally linked, the contractile and the supportive matrix structures upregulate their protein synthesis rate quite differently in response to feeding and contractile activity and intensity. PMID- 19903867 TI - Detecting children with arithmetic disabilities from kindergarten: evidence from a 3-year longitudinal study on the role of preparatory arithmetic abilities. AB - In a 3-year longitudinal study, 471 children were classified, based on their performances on arithmetic tests in first and second grade, as having persistent arithmetic disabilities (AD), persistent low achieving (LA), persistent typical achieving, inconsistent arithmetic disabilities (DF1), or inconsistent low achieving in arithmetic. Significant differences in the performances on the magnitude comparison in kindergarten (at age 5-6) were found between the AD and LA and between the AD and DF1 groups. Furthermore, the percentage of true positive AD children (at age 7-8) correctly diagnosed in kindergarten by combination of procedural counting, conceptual counting, and magnitude comparison tasks was 87.50%. When composing clinical samples, researchers should pay attention when stipulating restrictive or lenient cutoffs for arithmetic disabilities and select children based on their scores in 2 consecutive years, because the results of studies on persistent low achievers or children with inconsistent disabilities cannot be generalized to children with persistent arithmetic disabilities. PMID- 19903869 TI - Rice MicroRNA effector complexes and targets. AB - MicroRNAs (miRNAs) are small silencing RNAs with regulatory roles in gene expression. miRNAs interact with Argonaute (AGO) proteins to form effector complexes that cleave target mRNAs or repress translation. Rice (Oryza sativa) encodes four AGO1 homologs (AGO1a, AGO1b, AGO1c, and AGO1d). We used RNA interference (RNAi) to knock down the four AGO1s. The RNAi lines displayed pleiotropic developmental phenotypes and had increased accumulation of miRNA targets. AGO1a, AGO1b, and AGO1c complexes were purified and further characterized. The three AGO1s all have a strong preference for binding small RNAs (sRNAs) with 5' U and have Slicer activity. We cataloged the sRNAs in each AGO1 complex by deep sequencing and found that all three AGO1s predominantly bound known miRNAs. Most of the miRNAs were evenly distributed in the three AGO1 complexes, suggesting a redundant role for the AGO1s. Intriguingly, a subset of miRNAs were specifically incorporated into or excluded from one of the AGO1s, suggesting functional specialization among the AGO1s. Furthermore, we identified rice miRNA targets at a global level. The validated targets include transcription factors that control major stages of development and also genes involved in a variety of physiological processes, indicating a broad regulatory role for miRNAs in rice. PMID- 19903870 TI - Efficient operation of NAD(P)H dehydrogenase requires supercomplex formation with photosystem I via minor LHCI in Arabidopsis. AB - In higher plants, the chloroplast NAD(P)H dehydrogenase (NDH) complex mediates photosystem I (PSI) cyclic and chlororespiratory electron transport. We reported previously that NDH interacts with the PSI complex to form a supercomplex (NDH PSI). In this study, NDH18 and FKBP16-2 (FK506 Binding Protein 16-2), detected in the NDH-PSI supercomplex by mass spectrometry, were shown to be NDH subunits by the analysis of their knockdown lines. On the basis of extensive mutant characterization, we propose a structural model for chloroplast NDH, whereby NDH is divided into four subcomplexes. The subcomplex A and membrane subcomplex are conserved in cyanobacteria, but the subcomplex B and lumen subcomplex are specific to chloroplasts. Two minor light-harvesting complex I proteins, Lhca5 and Lhca6, were required for the full-size NDH-PSI supercomplex formation. Similar to crr pgr5 double mutants that completely lack cyclic electron flow activity around PSI, the lhca6 pgr5 double mutant exhibited a severe defect in growth. Consistent with the impaired NDH activity, photosynthesis was also severely affected in mature leaves of lhca6 pgr5. We conclude that chloroplast NDH became equipped with the novel subcomplexes and became associated with PSI during the evolution of land plants, and this process may have facilitated the efficient operation of NDH. PMID- 19903871 TI - Silver ions increase auxin efflux independently of effects on ethylene response. AB - Silver nitrate and aminoethoxyvinylglycine (AVG) are often used to inhibit perception and biosynthesis, respectively, of the phytohormone ethylene. In the course of exploring the genetic basis of the extensive interactions between ethylene and auxin, we compared the effects of silver nitrate (AgNO(3)) and AVG on auxin responsiveness. We found that although AgNO(3) dramatically decreased root indole-3-acetic acid (IAA) responsiveness in inhibition of root elongation, promotion of DR5-beta-glucuronidase activity, and reduction of Aux/IAA protein levels, AVG had more mild effects. Moreover, we found that that silver ions, but not AVG, enhanced IAA efflux similarly in root tips of both the wild type and mutants with blocked ethylene responses, indicating that this enhancement was independent of ethylene signaling. Our results suggest that the promotion of IAA efflux by silver ions is independent of the effects of silver ions on ethylene perception. Although the molecular details of this enhancement remain unknown, our finding that silver ions can promote IAA efflux in addition to blocking ethylene signaling suggest that caution is warranted in interpreting studies using AgNO(3) to block ethylene signaling in roots. PMID- 19903872 TI - Challenges to optimal enteral nutrition in a multidisciplinary pediatric intensive care unit. AB - OBJECTIVE: To describe nutrient intake in critically ill children, identify risk factors associated with avoidable interruptions to enteral nutrition (EN), and highlight opportunities to improve enteral nutrient delivery in a busy tertiary pediatric intensive care unit (PICU). Design, Setting, and Measurements: Daily nutrient intake and factors responsible for avoidable interruptions to EN were recorded in patients admitted to a 29-bed medical and surgical PICU over 4 weeks. Clinical characteristics, time to reach caloric goal, and parenteral nutrition (PN) use were compared between patients with and without avoidable interruptions to EN. RESULTS: Daily record of nutrient intake was obtained in 117 consecutive patients (median age, 7 years). Eighty (68%) patients received EN (20% postpyloric) for a total of 381 EN days (median, 2 days). Median time to EN initiation was less than 1 day. However, EN was subsequently interrupted in 24 (30%) patients at an average of 3.7 +/- 3.1 times per patient (range, 1-13), for a total of 88 episodes accounting for 1,483 hours of EN deprivation in this cohort. Of the 88 episodes of EN interruption, 51 (58%) were deemed as avoidable. Mechanically ventilated subjects were at the highest risk of EN interruptions. Avoidable EN interruption was associated with increased reliance on PN and impaired ability to reach caloric goal. CONCLUSIONS: EN interruption is common and frequently avoidable in critically ill children. Knowledge of existing barriers to EN such as those identified in this study will allow appropriate interventions to optimize nutrition provision in the PICU. PMID- 19903873 TI - Nonclinical safety assessment of the histone deacetylase inhibitor vorinostat. AB - Vorinostat (SAHA, Zolinza), a histone deacetylase inhibitor, is assessed in nonclinical studies to support its approval for cutaneous T-cell lymphoma. Vorinostat is weakly mutagenic in the Ames assay; is clastogenic in rodent (ie, CHO) cells but not in normal human lymphocytes; and is weakly positive in an in vivo mouse micronucleus assay. No effects are observed on potassium ion currents in the hERG assay up to 300 microM (safety margin approximately 300-fold the approximately 1 microM serum concentration associated with the 400 mg/d maximum recommended human dose. No rat respiratory or central nervous system effects are found at 150 mg/kg (>2-fold maximum recommended human dose). No cardiovascular effects, including effects on QTc interval, are observed after a single oral dose (150 mg/kg) in dogs. Vorinostat is orally dosed daily in rats (controls, 20, 50, or 150 mg/kg/d) and dogs (controls, 60, 80, or 100/125/160 mg/kg/d) for 26 weeks with a 4-week recovery. Rat vorinostat-related adverse findings are decreased food consumption, weight loss, and hematologic changes; a no observed adverse effects level is not established. In dogs, adverse effects are primarily gastrointestinal; the no observed adverse effects level is 60 mg/kg/d (approximately 6-fold maximum recommended human dose). Toxicities are reversible and can be monitored in the clinic. PMID- 19903874 TI - Deficits in spatial memory correlate with modified {gamma}-aminobutyric acid type A receptor tyrosine phosphorylation in the hippocampus. AB - Fast synaptic inhibition in the brain is largely mediated by gamma-aminobutyric acid receptors (GABA(A)R). While the pharmacological manipulation of GABA(A)R function by therapeutic agents, such as benzodiazepines can have profound effects on neuronal excitation and behavior, the endogenous mechanisms neurons use to regulate the efficacy of synaptic inhibition and their impact on behavior remains poorly understood. To address this issue, we created a knock-in mouse in which tyrosine phosphorylation of the GABA(A)Rs gamma2 subunit, a posttranslational modification that is critical for their functional modulation, has been ablated. These animals exhibited enhanced GABA(A)R accumulation at postsynaptic inhibitory synaptic specializations on pyramidal neurons within the CA3 subdomain of the hippocampus, primarily due to aberrant trafficking within the endocytic pathway. This enhanced inhibition correlated with a specific deficit in spatial object recognition, a behavioral paradigm dependent upon CA3. Thus, phospho-dependent regulation of GABA(A)R function involving just two tyrosine residues in the gamma2 subunit provides an input-specific mechanism that not only regulates the efficacy of synaptic inhibition, but has behavioral consequences. PMID- 19903875 TI - Intramolecular amide bonds stabilize pili on the surface of bacilli. AB - Gram-positive bacteria elaborate pili and do so without the participation of folding chaperones or disulfide bond catalysts. Sortases, enzymes that cut pilin precursors, form covalent bonds that link pilin subunits and assemble pili on the bacterial surface. We determined the x-ray structure of BcpA, the major pilin subunit of Bacillus cereus. The BcpA precursor encompasses 2 Ig folds (CNA(2) and CNA(3)) and one jelly-roll domain (XNA) each of which synthesizes a single intramolecular amide bond. A fourth amide bond, derived from the Ig fold of CNA(1), is formed only after pilin subunits have been incorporated into pili. We report that the domains of pilin precursors have evolved to synthesize a discrete sequence of intramolecular amide bonds, thereby conferring structural stability and protease resistance to pili. PMID- 19903876 TI - Bringing the Hutchinsonian niche into the 21st century: ecological and evolutionary perspectives. AB - G. Evelyn Hutchinson more than a half century ago proposed that one could characterize the ecological niche of a species as an abstract mapping of population dynamics onto an environmental space, the axes of which are abiotic and biotic factors that influence birth and death rates. If a habitat has conditions within a species' niche, a population should persist without immigration from external sources, whereas if conditions are outside the niche, it faces extinction. Analyses of species' niches are essential to understanding controls on species' geographical range limits and how these limits might shift in our rapidly changing world. Recent developments in ecology and evolutionary biology suggest it is time to revisit and refine Hutchinson's niche concept. After reviewing techniques for quantifying niches, I examine subtleties that arise because of impacts species have on their own environments, the density dependent modulation of how individuals experience environments, and the interplay of dispersal and temporal heterogeneity in determining population persistence. Moreover, the evolutionary record over all time scales reveals a spectrum of rates of change in species' niches, from rapid niche evolution to profound niche conservatism. Substantial challenges revolving around the evolutionary dimension of the Hutchinsonian niche include quantifying the magnitude of evolved intraspecific and clade-level variation in niches and understanding the factors that govern where along the spectrum of potential evolutionary rates any given lineage lies. A growing body of theory provides elements of a conceptual framework for understanding niche conservatism and evolution, paving the way for an evolutionary theory of the niche. PMID- 19903877 TI - Precuneus shares intrinsic functional architecture in humans and monkeys. AB - Evidence from macaque monkey tracing studies suggests connectivity-based subdivisions within the precuneus, offering predictions for similar subdivisions in the human. Here we present functional connectivity analyses of this region using resting-state functional MRI data collected from both humans and macaque monkeys. Three distinct patterns of functional connectivity were demonstrated within the precuneus of both species, with each subdivision suggesting a discrete functional role: (i) the anterior precuneus, functionally connected with the superior parietal cortex, paracentral lobule, and motor cortex, suggesting a sensorimotor region; (ii) the central precuneus, functionally connected to the dorsolateral prefrontal, dorsomedial prefrontal, and multimodal lateral inferior parietal cortex, suggesting a cognitive/associative region; and (iii) the posterior precuneus, displaying functional connectivity with adjacent visual cortical regions. These functional connectivity patterns were differentiated from the more ventral networks associated with the posterior cingulate, which connected with limbic structures such as the medial temporal cortex, dorsal and ventromedial prefrontal regions, posterior lateral inferior parietal regions, and the lateral temporal cortex. Our findings are consistent with predictions from anatomical tracer studies in the monkey, and provide support that resting-state functional connectivity (RSFC) may in part reflect underlying anatomy. These subdivisions within the precuneus suggest that neuroimaging studies will benefit from treating this region as anatomically (and thus functionally) heterogeneous. Furthermore, the consistency between functional connectivity networks in monkeys and humans provides support for RSFC as a viable tool for addressing cross species comparisons of functional neuroanatomy. PMID- 19903878 TI - Viscosity of glass-forming liquids. AB - The low-temperature dynamics of ultraviscous liquids hold the key to understanding the nature of glass transition and relaxation phenomena, including the potential existence of an ideal thermodynamic glass transition. Unfortunately, existing viscosity models, such as the Vogel-Fulcher-Tammann (VFT) and Avramov-Milchev (AM) equations, exhibit systematic error when extrapolating to low temperatures. We present a model offering an improved description of the viscosity-temperature relationship for both inorganic and organic liquids using the same number of parameters as VFT and AM. The model has a clear physical foundation based on the temperature dependence of configurational entropy, and it offers an accurate prediction of low-temperature isokoms without any singularity at finite temperature. Our results cast doubt on the existence of a Kauzmann entropy catastrophe and associated ideal glass transition. PMID- 19903879 TI - MRPS18-2 protein immortalizes primary rat embryonic fibroblasts and endows them with stem cell-like properties. AB - We report that the overexpression of mitochondrial ribosomal protein MRPS18-2 (S18-2) can immortalize primary rat embryonic fibroblasts (REFs). The immortalized cells (18IM) lose contact inhibition, form foci, and are capable of anchorage-independent growth. Concurrently, mesodermal markers, such as vimentin, smooth muscle actin, and Fut4, disappear completely. 18IM cells express embryonic stem cell markers, such as SSEA-1, Sox2, and Oct3/4. In confluent cultures, a portion of cells also express ectoderm- and endoderm-specific pan-keratin, ectoderm-specific beta-III-tubulin, mesoderm-specific MHC class II, and become stainable for fat with Oil red O. None of these changes was detected in c-myc+Ha ras (MR)-transformed cells. In immunodeficient mice, 18IM cells formed small transiently growing tumors that have down-regulated SSEA-1 and showed pan-keratin staining. We conclude that S18-2 can immortalize REFs and induces them to express stem cell traits. PMID- 19903880 TI - Her6 regulates the neurogenetic gradient and neuronal identity in the thalamus. AB - During vertebrate brain development, the onset of neuronal differentiation is under strict temporal control. In the mammalian thalamus and other brain regions, neurogenesis is regulated also in a spatially progressive manner referred to as a neurogenetic gradient, the underlying mechanism of which is unknown. Here we describe the existence of a neurogenetic gradient in the zebrafish thalamus and show that the progression of neurogenesis is controlled by dynamic expression of the bHLH repressor her6. Members of the Hes/Her family are known to regulate proneural genes, such as Neurogenin and Ascl. Here we find that Her6 determines not only the onset of neurogenesis but also the identity of thalamic neurons, marked by proneural and neurotransmitter gene expression: loss of Her6 leads to premature Neurogenin1-mediated genesis of glutamatergic (excitatory) neurons, whereas maintenance of Her6 leads to Ascl1-mediated production of GABAergic (inhibitory) neurons. Thus, the presence or absence of a single upstream regulator of proneural gene expression, Her6, leads to the establishment of discrete neuronal domains in the thalamus. PMID- 19903881 TI - Three-dimensional EM structure of an intact activator-dependent transcription initiation complex. AB - We present the experimentally determined 3D structure of an intact activator dependent transcription initiation complex comprising the Escherichia coli catabolite activator protein (CAP), RNA polymerase holoenzyme (RNAP), and a DNA fragment containing positions -78 to +20 of a Class I CAP-dependent promoter with a CAP site at position -61.5 and a premelted transcription bubble. A 20-A electron microscopy reconstruction was obtained by iterative projection-based matching of single particles visualized in carbon-sandwich negative stain and was fitted using atomic coordinate sets for CAP, RNAP, and DNA. The structure defines the organization of a Class I CAP-RNAP-promoter complex and supports previously proposed interactions of CAP with RNAP alpha subunit C-terminal domain (alphaCTD), interactions of alphaCTD with sigma(70) region 4, interactions of CAP and RNAP with promoter DNA, and phased-DNA-bend-dependent partial wrapping of DNA around the complex. The structure also reveals the positions and shapes of species-specific domains within the RNAP beta', beta, and sigma(70) subunits. PMID- 19903882 TI - Toward a quantitative theory of intrinsically disordered proteins and their function. AB - A large number of proteins are sufficiently unstable that their full 3D structure cannot be resolved. The origins of this intrinsic disorder are not well understood, but its ubiquitous presence undercuts the principle that a protein's structure determines its function. Here we present a quantitative theory that makes predictions regarding the role of intrinsic disorder in protein structure and function. In particular, we discuss the implications of analytical solutions of a series of fundamental thermodynamic models of protein interactions in which disordered proteins are characterized by positive folding free energies. We validate our predictions by assigning protein function by using the gene ontology classification--in which "protein binding", "catalytic activity", and "transcription regulator activity" are the three largest functional categories- and by performing genome-wide surveys of both the amount of disorder in these functional classes and binding affinities for both prokaryotic and eukaryotic genomes. Specifically, without assuming any a priori structure-function relationship, the theory predicts that both catalytic and low-affinity binding (K(d) greater, >or= 0(-7) M) proteins prefer ordered structures, whereas only high-affinity binding proteins (found mostly in eukaryotes) can tolerate disorder. Relevant to both transcription and signal transduction, the theory also explains how increasing disorder can tune the binding affinity to maximize the specificity of promiscuous interactions. Collectively, these studies provide insight into how natural selection acts on folding stability to optimize protein function. PMID- 19903883 TI - Reprogramming erythroid cells for lysosomal enzyme production leads to visceral and CNS cross-correction in mice with Hurler syndrome. AB - Restricting transgene expression to maturing erythroid cells can reduce the risk for activating oncogenes in hematopoietic stem cells (HSCs) and their progeny, yet take advantage of their robust protein synthesis machinery for high-level protein production. This study sought to evaluate the feasibility and efficacy of reprogramming erythroid cells for production of a lysosomal enzyme, alpha-L iduronidase (IDUA). An erythroid-specific hybrid promoter provided inducible IDUA expression and release during in vitro erythroid differentiation in murine erythroleukemia cells, resulting in phenotypical cross-correction in an enzyme deficient lymphoblastoid cell line derived from patients with mucopolysaccharidosis type I (MPS I). Stable and higher than normal plasma IDUA levels were achieved in vivo in primary and secondary MPS I chimeras for at least 9 months after transplantation of HSCs transduced with the erythroid-specific IDUA-containing lentiviral vector (LV). Moreover, long-term metabolic correction was demonstrated by normalized urinary glycosaminoglycan accumulation in all treated MPS I mice. Complete normalization of tissue pathology was observed in heart, liver, and spleen. Notably, neurological function and brain pathology were significantly improved in MPS I mice by erythroid-derived, higher than normal peripheral IDUA protein. These data demonstrate that late-stage erythroid cells, transduced with a tissue-specific LV, can deliver a lysosomal enzyme continuously at supraphysiological levels to the bloodstream and can correct the disease phenotype in both viscera and CNS of MPS I mice. This approach provides a paradigm for the utilization of RBC precursors as a depot for efficient and potentially safer systemic delivery of nonsecreted proteins by ex vivo HSC gene transfer. PMID- 19903884 TI - Single molecule detection of direct, homologous, DNA/DNA pairing. AB - Using a parallel single molecule magnetic tweezers assay we demonstrate homologous pairing of two double-stranded (ds) DNA molecules in the absence of proteins, divalent metal ions, crowding agents, or free DNA ends. Pairing is accurate and rapid under physiological conditions of temperature and monovalent salt, even at DNA molecule concentrations orders of magnitude below those found in vivo, and in the presence of a large excess of nonspecific competitor DNA. Crowding agents further increase the reaction rate. Pairing is readily detected between regions of homology of 5 kb or more. Detected pairs are stable against thermal forces and shear forces up to 10 pN. These results strongly suggest that direct recognition of homology between chemically intact B-DNA molecules should be possible in vivo. The robustness of the observed signal raises the possibility that pairing might even be the "default" option, limited to desired situations by specific features. Protein-independent homologous pairing of intact dsDNA has been predicted theoretically, but further studies are needed to determine whether existing theories fit sequence length, temperature, and salt dependencies described here. PMID- 19903885 TI - Identification of CCR4 and other essential thyroid hormone receptor co-activators by modified yeast synthetic genetic array analysis. AB - Identification of thyroid hormone receptor (TR) co-regulators has enhanced our understanding of thyroid hormone (TH) action. However, it is likely that many other co-regulators remained unidentified, and unbiased methods are required to discover these proteins. We have previously demonstrated that the yeast Saccharomyces cerevisiae is an excellent system in which to study TR action, and that defined TR signaling complexes in a eukaryotic background devoid of complicating influences of mammalian cell co-regulators can be constructed and analyzed for endogenous yeast genes, many of which are conserved in mammals. Here, a modified synthetic genetic array analysis was performed by crossing a yeast strain that expressed TRbeta1 and the co-activator GRIP1/SRC2 with 384 yeast strains bearing deletions of known genes. Eight genes essential for TH action were isolated, of which 4 are conserved in mammals. Examination of one, the yeast CCR4 and its human homolog CCR4/NOT6 (hCCR4), confirmed that (i) transfected CCR4 potentiates a TH response in cultured cells more efficiently than established TR co-activators and (ii) knockdown of CCR4 expression strongly inhibited a TH response (>80%). TH treatment promoted rapid and sustained hCCR4 recruitment to the TH-responsive deiodinase 1 promoter and TR co-localizes with hCCR4 in the nucleus and interacts with hCCR4 in 2-hybrid and pull-down assays. These findings indicate that a modified yeast synthetic genetic array strategy is a feasible method for unbiased identification of conserved genes essential for TR and other nuclear receptor hormone functions in mammals. PMID- 19903886 TI - Two size-selective mechanisms specifically trap bacteria-sized food particles in Caenorhabditis elegans. AB - Caenorhabditis elegans is a filter feeder: it draws bacteria suspended in liquid into its pharynx, traps the bacteria, and ejects the liquid. How pharyngeal pumping simultaneously transports and filters food particles has been poorly understood. Here, we use high-speed video microscopy to define the detailed workings of pharyngeal mechanics. The buccal cavity and metastomal flaps regulate the flow of dense bacterial suspensions and exclude excessively large particles from entering the pharynx. A complex sequence of contractions and relaxations transports food particles in two successive trap stages before passage into the terminal bulb and intestine. Filtering occurs at each trap as bacteria are concentrated in the central lumen while fluids are expelled radially through three apical channels. Experiments with microspheres show that the C. elegans pharynx, in combination with the buccal cavity, is tuned to specifically catch and transport particles of a size range corresponding to most soil bacteria. PMID- 19903887 TI - Localization of mRNAs coding for peroxisomal proteins in the yeast, Saccharomyces cerevisiae. AB - Targeted mRNA trafficking and local translation may play a significant role in controlling protein localization. Here we examined for the first time the localization of all ( approximately 50) mRNAs encoding peroxisomal proteins (mPPs) involved in peroxisome biogenesis and function. By using the bacteriophage MS2-CP RNA-binding protein (RBP) fused to multiple copies of GFP, we demonstrated that >40 endogenously expressed mPPs tagged with the MS2 aptamer form fluorescent RNA granules in vivo. The use of different RFP-tagged organellar markers revealed 3 basic patterns of mPP granule localization: to peroxisomes, to the endoplasmic reticulum (ER), and nonperoxisomal. Twelve mPPs (i.e., PEX1, PEX5, PEX8, PEX11 15, DCI1, NPY1, PCS60, and POX1) had a high percentage (52%-80%) of mRNA colocalization with peroxisomes. Thirteen mPPs (i.e., AAT2, PEX6, MDH3, PEX28, etc.) showed a low percentage (30%-42%) of colocalization, and 1 mPP (PEX3) preferentially localized to the ER. The mPPs of the nonperoxisomal pattern (i.e., GPD1, PCD1, PEX7) showed <<30% colocalization. mPP association with the peroxisome or ER was verified using cell fractionation and RT-PCR analysis. A model mPP, PEX14 mRNA, was found to be in close association with peroxisomes throughout the cell cycle, with its localization depending in part on the 3'-UTR, initiation of translation, and the Puf5 RBP. The different patterns of mPP localization observed suggest that multiple mechanisms involved in mRNA localization and translation may play roles in the importation of protein into peroxisomes. PMID- 19903888 TI - DAB2IP coordinates both PI3K-Akt and ASK1 pathways for cell survival and apoptosis. AB - In metastatic prostate cancer (PCa) cells, imbalance between cell survival and death signals such as constitutive activation of phosphatidylinositol 3-kinase (PI3K)-Akt and inactivation of apoptosis-stimulated kinase (ASK1)-JNK pathways is often detected. Here, we show that DAB2IP protein, often down-regulated in PCa, is a potent growth inhibitor by inducing G(0)/G(1) cell cycle arrest and is proapoptotic in response to stress. Gain of function study showed that DAB2IP can suppress the PI3K-Akt pathway and enhance ASK1 activation leading to cell apoptosis, whereas loss of DAB2IP expression resulted in PI3K-Akt activation and ASK1-JNK inactivation leading to accelerated PCa growth in vivo. Moreover, glandular epithelia from DAB2IP(-/-) animal exhibited hyperplasia and apoptotic defect. Structural functional analyses of DAB2IP protein indicate that both proline-rich (PR) and PERIOD-like (PER) domains, in addition to the critical role of C2 domain in ASK1 activity, are important for modulating PI3K-Akt activity. Thus, DAB2IP is a scaffold protein capable of bridging both survival and death signal molecules, which implies its role in maintaining cell homeostasis. PMID- 19903893 TI - Abstracts of the Gerontological Society of America (GSA) 62nd Annual Scientific Meeting. November 18-22, 2009. Atlanta, Georgia, USA. PMID- 19903889 TI - Production of hyperpolarized [1,4-13C2]malate from [1,4-13C2]fumarate is a marker of cell necrosis and treatment response in tumors. AB - Dynamic nuclear polarization of (13)C-labeled cell substrates has been shown to massively increase their sensitivity to detection in NMR experiments. The sensitivity gain is sufficiently large that if these polarized molecules are injected intravenously, their spatial distribution and subsequent conversion into other cell metabolites can be imaged. We have used this method to image the conversion of fumarate to malate in a murine lymphoma tumor in vivo after i.v. injection of hyperpolarized [1,4-(13)C(2)]fumarate. In isolated lymphoma cells, the rate of labeled malate production was unaffected by coadministration of succinate, which competes with fumarate for transport into the cell. There was, however, a correlation with the percentage of cells that had lost plasma membrane integrity, suggesting that the production of labeled malate from fumarate is a sensitive marker of cellular necrosis. Twenty-four hours after treating implanted lymphoma tumors with etoposide, at which point there were significant levels of tumor cell necrosis, there was a 2.4-fold increase in hyperpolarized [1,4 (13)C(2)]malate production compared with the untreated tumors. Therefore, the formation of hyperpolarized (13)C-labeled malate from [1,4-(13)C(2)]fumarate appears to be a sensitive marker of tumor cell death in vivo and could be used to detect the early response of tumors to treatment. Given that fumarate is an endogenous molecule, this technique has the potential to be used clinically. PMID- 19903896 TI - ALK1 signaling regulates early postnatal lymphatic vessel development. AB - In vertebrates, endothelial cells form 2 hierarchical tubular networks, the blood vessels and the lymphatic vessels. Despite the difference in their structure and function and genetic programs that dictate their morphogenesis, common signaling pathways have been recognized that regulate both vascular systems. ALK1 is a member of the transforming growth factor-beta type I family of receptors, and compelling genetic evidence suggests its essential role in regulating blood vascular development. Here we report that ALK1 signaling is intimately involved in lymphatic development. Lymphatic endothelial cells express key components of the ALK1 pathway and respond robustly to ALK1 ligand stimulation in vitro. Blockade of ALK1 signaling results in defective lymphatic development in multiple organs of neonatal mice. We find that ALK1 signaling regulates the differentiation of lymphatic endothelial cells to influence the lymphatic vascular development and remodeling. Furthermore, simultaneous inhibition of ALK1 pathway increases apoptosis in lymphatic vessels caused by blockade of VEGFR3 signaling. Thus, our study reveals a novel aspect of ALK1 signaling in regulating lymphatic development and suggests that targeting ALK1 pathway might provide additional control of lymphangiogenesis in human diseases. PMID- 19903895 TI - Memories that last forever: strategies for optimizing vaccine T-cell memory. AB - For acute self-limiting infections a vaccine is successful if it elicits memory at least as good as the natural experience; however, for persistent and chronic infections such as HIV, hepatitis C virus (HCV), human papillomavirus (HPV), and human herpes viruses, this paradigm is not applicable. At best, during persistent virus infection the person must be able to maintain the integrity of the immune system in equilibrium with controlling replicating virus. New vaccine strategies are required that elicit both potent high-avidity CD8(+) T-cell effector/memory and central memory responses that can clear the nidus of initial virus-infected cells at mucosal surfaces to prevent mucosal transmission or significantly curtail development of disease. The objective of an HIV-1 T-cell vaccine is to generate functional CD8(+) effector memory cells at mucosal portals of virus entry to prevent viral transmission. In addition, long-lived CD8(+) and CD4(+) central memory cells circulating through secondary lymphoid organs and resident in bone marrow, respectively, are needed to provide a concerted second wave of defense that can contain virus at mucosal surfaces and prevent systemic dissemination. Further understanding of factors which can influence long-lived effector and central memory cell differentiation will significantly contribute to development of effective T-cell vaccines. In this review we will focus on discussing mechanisms involved in T-cell memory and provide promising new approaches toward expanding current vaccine strategies to enhance antiviral memory. PMID- 19903897 TI - The effect of prolonged administration of hydroxyurea on morbidity and mortality in adult patients with sickle cell syndromes: results of a 17-year, single-center trial (LaSHS). AB - The aim of this prospective study was to evaluate the long-term efficacy and safety of hydroxyurea (HU) in patients with sickle cell disease (SCD). Thirty four patients with sickle cell anemia (hemoglobin S [HbS]/HbS), 131 with HbS/beta(0)-thal, and 165 with HbS/beta(+)-thal participated in this trial. HU was administered to 131 patients, whereas 199 patients were conventionally treated. The median follow-up period was 8 years for HU patients and 5 years for non-HU patients. HU produced a dramatic reduction in the frequency of severe painful crises, transfusion requirements, hospital admissions, and incidence of acute chest syndrome. The probability of 10-year survival was 86% and 65% for HU and non-HU patients, respectively (P = .001), although HU patients had more severe forms of SCD. The 10-year probability of survival for HbS/HbS, HbS/beta (0)-thal, and HbS/IVSI-110 patients was 100%, 87%, and 82%, respectively, for HU patients and 10%, 54%, and 66%, for non-HU patients. The multivariate analysis showed that fetal hemoglobin values at baseline and percentage change of lactate dehydrogenase between baseline and 6 months were independently predicted for survival in the HU group. These results highlight the beneficial effect of HU, which seems to modify the natural history of SCD and raise the issue of expanding its use in all SCD patients. PMID- 19903898 TI - Quantitative DNA methylation predicts survival in adult acute myeloid leukemia. AB - Acute myeloid leukemia (AML) is characterized by molecular heterogeneity that is not fully reflected in the current classification system. Recent insights point toward a significant role of aberrant DNA methylation in leukemogenesis. Therefore, we investigated the prognostic impact of DNA methylation in AML. To screen for promoter methylation in AML we applied a combination of base-specific cleavage biochemistry and matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF-MS), a powerful methodology allowing for quantitatively investigating DNA methylation status in a large series of both promoter regions and leukemia samples. We analyzed 92 genomic regions in 182 patient samples, correlated findings with clinical and molecular data, and validated the results in an independent cohort of 74 AML samples. Using this approach, we were able to identify novel leukemia subgroups based on distinct DNA methylation patterns. Furthermore, we defined a methylation-based outcome predictor for patient survival (P < .01) that in multivariable analysis provided independent prognostic information (hazard ratio, 1.52; 95% CI, 1.06-2.16). Here, we report the first large-scale methylation-based outcome predictor in AML, and thereby our findings support the use of genomic methylation markers for improved molecular classification and prognostication in adult AML. PMID- 19903899 TI - Galectin-5 is bound onto the surface of rat reticulocyte exosomes and modulates vesicle uptake by macrophages. AB - Reticulocytes release small membrane vesicles termed exosomes during their maturation into erythrocytes. Exosomes are intraluminal vesicles of multivesicular endosomes released into the extracellular medium by fusion of these endosomal compartments with the plasma membrane. This secretion pathway contributes to reticulocyte plasma membrane remodeling by eliminating certain membrane glycoproteins. We show in this study that galectin-5, although mainly cytosolic, is also present on the cell surface of rat reticulocytes and erythrocytes. In addition, in reticulocytes, it resides in the endosomal compartment. We document galectin-5 translocation from the cytosol into the endosome lumen, leading to its secretion in association with exosomes. Galectin-5 bound onto the vesicle surface may function in sorting galactose-bearing glycoconjugates. Fittingly, we found that Lamp2, a major cellular glycoprotein presenting galectin-reactive poly-N-acetylactosamine chains, is lost during reticulocyte maturation. It is associated with released exosomes, suggestive of binding to galectin-5. Finally, we reveal that the uptake of rat reticulocyte exosomes by macrophages is dependent on temperature and the mechanoenzyme dynamin and that exosome uptake is decreased by adding galectin-5. These data imply galectin-5 functionality in the exosomal sorting pathway during rat reticulocyte maturation. PMID- 19903900 TI - Education of human natural killer cells by activating killer cell immunoglobulin like receptors. AB - Expression of inhibitory killer cell immunoglobulin-like receptors (KIRs) specific for self-major histocompatibility complex (MHC) class I molecules provides an educational signal that generates functional natural killer (NK) cells. However, the effects of activating KIRs specific for self-MHC class I on NK-cell education remain elusive. Here, we provide evidence that the activating receptor KIR2DS1 tunes down the responsiveness of freshly isolated human NK cells to target cell stimulation in donors homozygous for human leukocyte antigen (HLA) C2, the ligand of KIR2DS1. The tuning was apparent in KIR2DS1(+) NK cells lacking expression of inhibitory KIRs and CD94/NKG2A, as well as in KIR2DS1(+) NK cells coexpressing the inhibitory MHC class I-specific receptors CD94/NKG2A and KIR2DL3, but not KIR2DL1. However, the tuning of responsiveness was restricted to target cell recognition because KIR2DS1(+) NK cells responded well to stimulation with exogenous cytokines. Our results provide the first example of human NK-cell education by an activating KIR and suggest that the education of NK cells via activating KIRs is a mechanism to secure tolerance that complements education via inhibitory KIRs. PMID- 19903901 TI - The role of the Th1 transcription factor T-bet in a mouse model of immune mediated bone-marrow failure. AB - The transcription factor T-bet is a key regulator of type 1 immune responses. We examined the role of T-bet in an animal model of immune-mediated bone marrow (BM) failure using mice carrying a germline T-bet gene deletion (T-bet(-/-)). In comparison with normal C57BL6 (B6) control mice, T-bet(-/-) mice had normal cellular composition in lymphohematopoietic tissues, but T-bet(-/-) lymphocytes were functionally defective. Infusion of 5 x 10(6) T-bet(-/-) lymph node (LN) cells into sublethally irradiated, major histocompatibility complex-mismatched CByB6F1 (F1) recipients failed to induce the severe marrow hypoplasia and fatal pancytopenia that is produced by injection of similar numbers of B6 LN cells. Increasing T-bet(-/-) LN-cell dose to 10 to 23 x 10(6) per recipient led to only mild hematopoietic deficiency. Recipients of T-bet(-/-) LN cells had no expansion in T cells or interferon-gamma-producing T cells but showed a significant increase in Lin(-)Sca1(+)CD117(+)CD34(-) BM cells. Plasma transforming growth factor-beta and interleukin-17 concentrations were increased in T-bet(-/-) LN cell recipients, possibly a compensatory up-regulation of the Th17 immune response. Continuous infusion of interferon-gamma resulted in hematopoietic suppression but did not cause T-bet(-/-) LN-cell expansion or BM destruction. Our data provided fresh evidence demonstrating a critical role of T-bet in immune mediated BM failure. PMID- 19903902 TI - In vivo biotinylation of the vasculature in B-cell lymphoma identifies BST-2 as a target for antibody-based therapy. AB - The discovery of accessible markers of lymphoma may facilitate the development of antibody-based therapeutic strategies. Here, we describe the results of a chemical proteomic study, based on the in vivo biotinylation of vascular proteins in lymphoma-bearing mice followed by mass spectrometric and bioinformatic analysis, to discover proteins expressed at the tissue-blood border of disseminated B-cell lymphoma. From a list of 58 proteins, which were more than 10 fold up-regulated in nodal and extranodal lymphoma lesions compared with their levels in the corresponding normal host organs, we validated BST-2 as a novel vascular marker of B-cell lymphoma, using immunochemical techniques and in vivo biodistribution studies. Furthermore, targeting BST-2 with 2 independent monoclonal antibodies delayed lymphoma growth in a syngeneic mouse model of the disease. The results of this study delineate a strategy for the treatment of systemic B-cell lymphoma in humans and suggest that anti-BST-2 antibodies may facilitate pharmacodelivery approaches that target the tumor-stroma interface. PMID- 19903903 TI - Autoimmune B-cell lymphopenia after successful adoptive therapy with telomerase specific T lymphocytes. AB - Telomerase reverse transcriptase (TERT) is a good candidate for cancer immunotherapy because it is overexpressed in 85% of all human tumors and implicated in maintenance of the transformed phenotype. TERT-based cancer vaccines have been shown to be safe, not inducing any immune-related pathology, but their impact on tumor progression is modest. Here we show that adoptive cell therapy with the use of high-avidity T lymphocytes reactive against telomerase can control the growth of different established tumors. Moreover, in transgenic adenocarcinoma mouse prostate mice, which develop prostate cancer, TERT-based adoptive cell therapy halted the progression to more aggressive and poorly differentiated tumors, significantly prolonging mouse survival. We also demonstrated that human tumors, including Burkitt lymphoma, and human cancer stem cells, are targeted in vivo by TERT-specific cytotoxic T lymphocytes. Effective therapy with T cells against telomerase, different from active vaccination, however, led to autoimmunity marked by a consistent, although transient, B-cell depletion in primary and secondary lymphoid organs, associated with alteration of the spleen cytoarchitecture. These results indicate B cells as an in vivo target of TERT-specific cytotoxic T lymphocytes during successful immunotherapy. PMID- 19903904 TI - Failure to report financial disclosure information. PMID- 19903905 TI - Communicating with pictures: perceptions of cardiovascular health among Asian immigrants. PMID- 19903906 TI - The cover. Alma sewing. PMID- 19903907 TI - A piece of my mind. Death and life in Afghanistan. PMID- 19903908 TI - Silent epidemic of viral hepatitis may lead to boom in serious liver disease. PMID- 19903909 TI - New clinical guideline for hoarseness offers assessment and treatment advice. PMID- 19903910 TI - New standards reduce heparin potency. PMID- 19903912 TI - Outcomes in patients with prolonged PR interval or first-degree atrioventricular block. PMID- 19903913 TI - Outcomes in patients with prolonged PR interval or first-degree atrioventricular block. PMID- 19903914 TI - Polymyxin B hemoperfusion and mortality in abdominal septic shock. PMID- 19903915 TI - Polymyxin B hemoperfusion and mortality in abdominal septic shock. PMID- 19903916 TI - Polymyxin B hemoperfusion and mortality in abdominal septic shock. PMID- 19903917 TI - Estimate of the carbon footprint of the US health care sector. PMID- 19903918 TI - Prone positioning in patients with moderate and severe acute respiratory distress syndrome: a randomized controlled trial. AB - CONTEXT: Post hoc analysis of a previous trial has suggested that prone positioning may improve survival in patients with severe hypoxemia and with acute respiratory distress syndrome (ARDS). OBJECTIVE: To assess possible outcome benefits of prone positioning in patients with moderate and severe hypoxemia who are affected by ARDS. DESIGN, SETTING, AND PATIENTS: The Prone-Supine II Study, a multicenter, unblinded, randomized controlled trial conducted in 23 centers in Italy and 2 in Spain. Patients were 342 adults with ARDS receiving mechanical ventilation, enrolled from February 2004 through June 2008 and prospectively stratified into subgroups with moderate (n = 192) and severe (n = 150) hypoxemia. INTERVENTIONS: Patients were randomized to undergo supine (n = 174) or prone (20 hours per day; n = 168) positioning during ventilation. MAIN OUTCOME MEASURES: The primary outcome was 28-day all-cause mortality. Secondary outcomes were 6 month mortality and mortality at intensive care unit discharge, organ dysfunctions, and the complication rate related to prone positioning. RESULTS: Prone and supine patients from the entire study population had similar 28-day (31.0% vs 32.8%; relative risk [RR], 0.97; 95% confidence interval [CI], 0.84 1.13; P = .72) and 6-month (47.0% vs 52.3%; RR, 0.90; 95% CI, 0.73-1.11; P = .33) mortality rates, despite significantly higher complication rates in the prone group. Outcomes were also similar for patients with moderate hypoxemia in the prone and supine groups at 28 days (25.5% vs 22.5%; RR, 1.04; 95% CI, 0.89-1.22; P = .62) and at 6 months (42.6% vs 43.9%; RR, 0.98; 95% CI, 0.76-1.25; P = .85). The 28-day mortality of patients with severe hypoxemia was 37.8% in the prone and 46.1% in the supine group (RR, 0.87; 95% CI, 0.66-1.14; P = .31), while their 6 month mortality was 52.7% and 63.2%, respectively (RR, 0.78; 95% CI, 0.53-1.14; P = .19). CONCLUSION: Data from this study indicate that prone positioning does not provide significant survival benefit in patients with ARDS or in subgroups of patients with moderate and severe hypoxemia. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00159939. PMID- 19903919 TI - Prevalence of and factors associated with persistent pain following breast cancer surgery. AB - CONTEXT: Persistent pain and sensory disturbances following surgical treatment for breast cancer is a significant clinical problem. The pathogenic mechanisms are complex and may be related to patient characteristics, surgical technique, and adjuvant therapy. OBJECTIVE: To examine prevalence of and factors associated with persistent pain after surgical treatment for breast cancer. DESIGN, SETTING, AND PATIENTS: A nationwide cross-sectional questionnaire study of 3754 women aged 18 to 70 years who received surgery and adjuvant therapy (if indicated) for primary breast cancer in Denmark between January 1, 2005, and December 31, 2006. A study questionnaire was sent to the women between January and April 2008. MAIN OUTCOME MEASURES: Prevalence, location, and severity of persistent pain and sensory disturbances in 12 well-defined treatment groups assessed an average of 26 months after surgery, and adjusted odds ratio (OR) of reported pain and sensory disturbances with respect to age, surgical technique, chemotherapy, and radiotherapy. RESULTS: By June 2008, 3253 of 3754 eligible women (87%) returned the questionnaire. A total of 1543 patients (47%) reported pain, of whom 201 (13%) had severe pain, 595 (39%) had moderate pain, and 733 (48%) had light pain. Factors associated with chronic pain included young age (18-39 years: OR, 3.62; 95% confidence interval [CI], 2.25-5.82; P < .001) and adjuvant radiotherapy (OR, 1.50; 95% CI, 1.08-2.07; P = .03), but not chemotherapy (OR, 1.01; 95% CI, 0.85 1.21; P = .91). Axillary lymph node dissection (ALND) was associated with increased likelihood of pain (OR, 1.77; 95% CI, 1.43-2.19; P < .001) compared with sentinel lymph node dissection. Risk of sensory disturbances was associated with young age (18-39 years: OR, 5.00; 95% CI, 2.87-8.69; P < .001) and ALND (OR, 4.97; 95% CI, 3.92-6.30; P < .001). Pain complaints from other parts of the body were associated with increased risk of pain in the surgical area (P < .001). A total of 306 patients (20%) with pain had contacted a physician within the prior 3 months for pain complaints in the surgical area. CONCLUSION: Two to 3 years after breast cancer treatment, persistent pain and sensory disturbances remain clinically significant problems among Danish women who received surgery in 2005 and 2006. PMID- 19903921 TI - Statin use and risk of gallstone disease followed by cholecystectomy. AB - CONTEXT: Gallstone disease is a leading cause of morbidity in western countries and carries a high economic burden. Statins decrease hepatic cholesterol biosynthesis and may therefore lower the risk of cholesterol gallstones by reducing the cholesterol concentration in the bile. Data on this association in humans are scarce. OBJECTIVE: To study the association between the use of statins, fibrates, or other lipid-lowering agents and the risk of incident gallstone disease followed by cholecystectomy. DESIGN, SETTING, AND PARTICIPANTS: Case-control analysis using the UK-based General Practice Research Database. Incident patients between 1994 and 2008 and 4 controls per each patient were identified and matched on age, sex, general practice, calendar time, and years of history in the database. The study population was 76% women and the mean (SD) age was 53.4 (15.0) years at the index date. Conditional logistic regression was used to estimate the odds ratio (OR) of developing gallstones followed by cholecystectomy in relation to exposure to lipid-lowering agents, stratified by exposure timing and duration. The ORs and 95% confidence intervals (CIs) were adjusted for smoking, body mass index, ischemic heart disease, stroke, and estrogen use. MAIN OUTCOME MEASURE: The adjusted OR (AOR) for developing gallstone disease followed by cholecystectomy in relation to exposure to lipid lowering agents. RESULTS: A total of 27,035 patients with cholecystectomy and 106,531 matched controls were identified, including 2396 patients and 8868 controls who had statin use. Compared with nonuse, current statin use (last prescription recorded within 90 days before the first-time diagnosis of the disease) was 1.0% for patients and 0.8% for controls (AOR, 1.10; 95% CI, 0.95 1.27) for 1 to 4 prescriptions; 2.6% vs 2.4% (AOR, 0.85; 95% CI, 0.77-0.93) for 5 to 19 prescriptions, and 3.2% vs 3.7% (AOR, 0.64; 95% CI, 0.59-0.70) for 20 or more prescriptions. The AORs for current use of statins defined as 20 or more prescriptions were similar (around 0.6) across age, sex, and body mass index categories, and across the statin class. CONCLUSION: Long-term use of statins was associated with a decreased risk of gallstones followed by cholecystectomy. PMID- 19903922 TI - Shared medical regulation in a time of increasing calls for accountability and transparency: comparison of recertification in the United States, Canada, and the United Kingdom. AB - In the United States, Canada, and the United Kingdom, the medical profession is accountable to the public for the delivery and quality of care provided to patients. Traditionally, this accountability has been achieved through the development and maintenance of professional standards established by the profession itself-self-regulation. Medical self-regulation is being re-examined by regulators, government, and the profession in response to a range of drivers including payers seeking ways to hold physicians accountable for cost-effective care; patients seeking more information about their physician's qualifications; and the emergence of a number of high-profile cases of unacceptable medical practice. This article outlines the current state of medical regulation in the United States, Canada, and the United Kingdom and highlights the increasing external pressure on the self-regulatory framework that is leading to a shift toward shared regulation between the profession and other stakeholders. PMID- 19903923 TI - Update: A 55-year-old woman with osteopenia. PMID- 19903924 TI - Clinical research sites--the underappreciated component of the clinical research system. PMID- 19903925 TI - Genome-wide association studies and human disease: from trickle to flood. PMID- 19903926 TI - Improving outcomes in critically ill patients: the seduction of physiology. PMID- 19903927 TI - The purpose and limits to professional self-regulation. PMID- 19903920 TI - Major lipids, apolipoproteins, and risk of vascular disease. AB - CONTEXT: Associations of major lipids and apolipoproteins with the risk of vascular disease have not been reliably quantified. OBJECTIVE: To assess major lipids and apolipoproteins in vascular risk. DESIGN, SETTING, AND PARTICIPANTS: Individual records were supplied on 302,430 people without initial vascular disease from 68 long-term prospective studies, mostly in Europe and North America. During 2.79 million person-years of follow-up, there were 8857 nonfatal myocardial infarctions, 3928 coronary heart disease [CHD] deaths, 2534 ischemic strokes, 513 hemorrhagic strokes, and 2536 unclassified strokes. MAIN OUTCOME MEASURES: Hazard ratios (HRs), adjusted for several conventional factors, were calculated for 1-SD higher values: 0.52 log(e) triglyceride, 15 mg/dL high density lipoprotein cholesterol (HDL-C), 43 mg/dL non-HDL-C, 29 mg/dL apolipoprotein AI, 29 mg/dL apolipoprotein B, and 33 mg/dL directly measured low density lipoprotein cholesterol (LDL-C). Within-study regression analyses were adjusted for within-person variation and combined using meta-analysis. RESULTS: The rates of CHD per 1000 person-years in the bottom and top thirds of baseline lipid distributions, respectively, were 2.6 and 6.2 with triglyceride, 6.4 and 2.4 with HDL-C, and 2.3 and 6.7 with non-HDL-C. Adjusted HRs for CHD were 0.99 (95% CI, 0.94-1.05) with triglyceride, 0.78 (95% CI, 0.74-0.82) with HDL-C, and 1.50 (95% CI, 1.39-1.61) with non-HDL-C. Hazard ratios were at least as strong in participants who did not fast as in those who did. The HR for CHD was 0.35 (95% CI, 0.30-0.42) with a combination of 80 mg/dL lower non-HDL-C and 15 mg/dL higher HDL-C. For the subset with apolipoproteins or directly measured LDL-C, HRs were 1.50 (95% CI, 1.38-1.62) with the ratio non-HDL-C/HDL-C, 1.49 (95% CI, 1.39-1.60) with the ratio apo B/apo AI, 1.42 (95% CI, 1.06-1.91) with non-HDL-C, and 1.38 (95% CI, 1.09-1.73) with directly measured LDL-C. Hazard ratios for ischemic stroke were 1.02 (95% CI, 0.94-1.11) with triglyceride, 0.93 (95% CI, 0.84-1.02) with HDL-C, and 1.12 (95% CI, 1.04-1.20) with non-HDL-C. CONCLUSION: Lipid assessment in vascular disease can be simplified by measurement of either total and HDL cholesterol levels or apolipoproteins without the need to fast and without regard to triglyceride. PMID- 19903928 TI - Evaluating patients with chronic pain after breast cancer surgery: the search for relief. PMID- 19903929 TI - JAMA patient page. Aortic aneurysms. PMID- 19903930 TI - Pulse wave analysis of the aortic pressure waveform in severe left ventricular systolic dysfunction. AB - BACKGROUND: The effect of moderate left ventricular systolic dysfunction (LVSD) on ventricular/vascular coupling and the aortic pressure waveform (AoPW) has been well described, but the effect of severe LVSD has not. METHODS AND RESULTS: We used noninvasive, high-fidelity tonometry of the radial artery and a mathematical transfer function to generate the AoPW in 25 treated patients with LVSD (mean LV ejection fraction, 24+/-8.8%; range, 11% to 40%; 21 patients <30%). Pulse wave analysis of the AoPW was used to characterize ventricular/vascular coupling and compared with pulse wave analysis performed in 25 normal subjects matched for age, gender, height, body mass index, and heart rate. Measurements obtained using pulse wave analysis in LVSD patients indicated features of poor LV stroke performance and also reduced indices of arterial stiffness: increased travel time of the pressure wave (147+/-10 ms versus 132+/-21 ms; P<0.001); decreased systolic duration of reflected wave (134+/-24 ms versus 167+/-26 ms; P<0.001); ejection duration (277+/-22 ms versus 299+/-25 ms; P<0.008); percent systolic duration (32+/-5.3% versus 35+/-4.0%; P<0.02); aortic systolic pressure (100+/-16 mm Hg versus 121+/-16 mm Hg; P<0.001); unaugmented pressure (24+/-6.3 mm Hg versus 32+/-6.4 mm Hg; P<0.001); augmented pressure (4.8+/-3.1 mm Hg versus 9.6+/ 4.5 mm Hg; P<0.001); pulse pressure (28+/-7.4 mm Hg versus 42+/-9.5 mm Hg; P<0.001); augmentation index (12+/-6.6% versus 23+/-7.6%; P<0.006); wasted LV effort (5.3+/-2.8x10(2) dyne sec/cm(2) versus 17+/-10x10(2) dyne sec/cm(2); P<0.001); systolic pressure time index (17+/-4.1x10(2) mm Hg-sec/min versus 23+/ 4.2x10(2) mm Hg sec/min; P<0.001); and pressure systolic area (383+/-121 mm Hg sec/min versus 666+/-150 mm Hg sec/min; P<0.001). CONCLUSIONS: Severe LVSD causes measurable changes in the AoPW. Standardization of AoPW findings in LVSD patients may allow for the clinical use of radial artery pulse wave analysis to noninvasively determine the severity of dysfunction and aid in logical therapy. PMID- 19903931 TI - Recent national trends in readmission rates after heart failure hospitalization. AB - BACKGROUND: In July 2009, Medicare began publicly reporting hospitals' risk standardized 30-day all-cause readmission rates (RSRRs) among fee-for-service beneficiaries discharged after hospitalization for heart failure from all the US acute care nonfederal hospitals. No recent national trends in RSRRs have been reported, and it is not known whether hospital-specific performance is improving or variation in performance is decreasing. METHODS AND RESULTS: We used 2004-2006 Medicare administrative data to identify all fee-for-service beneficiaries admitted to a US acute care hospital for heart failure and discharged alive. We estimated mean annual RSRRs, a National Quality Forum-endorsed metric for quality, using 2-level hierarchical models that accounted for age, sex, and multiple comorbidities; variation in quality was estimated by the SD of the RSRRs. There were 570 996 distinct hospitalizations for heart failure in which the patient was discharged alive in 4728 hospitals in 2004, 544 550 in 4694 hospitals in 2005, and 501 234 in 4674 hospitals in 2006. Unadjusted 30-day all cause readmission rates were virtually identical over this period: 23.0% in 2004, 23.3% in 2005, and 22.9% in 2006. The mean and SD of RSRRs were also similar: mean (SD) of 23.7% (1.3) in 2004, 23.9% (1.4) in 2005, and 23.8% (1.4) in 2006, suggesting similar hospital variation throughout the study period. CONCLUSIONS: National mean and RSRR distributions among Medicare beneficiaries discharged after hospitalization for heart failure have not changed in recent years, indicating that there was neither improvement in hospital readmission rates nor in hospital variations in rates over this time period. PMID- 19903934 TI - Focus issue: coping with cellular stress. AB - Organisms constantly face potential damage from internal and external sources, thus necessitating signaling cascades that couple specific cellular stresses to the appropriate responses. This Focus Issue of Science Signaling highlights the signaling pathways that are activated by and that mediate responses to diverse types of stresses. PMID- 19903936 TI - Aiming straight for the heart: prolyl hydroxylases set the BAR. AB - The cellular response to a reduced oxygen state (or hypoxia) includes de novo alterations in gene expression patterns, many of which are controlled by hypoxia inducible factor (HIF) transcription factors. HIF signaling is predominantly regulated by the dioxygenase family of prolyl hydroxylases (PHDs), also known as EGL nine homologs (EGLNs). The PHD family in higher eukaryotes, like the HIF alpha family, is composed of multiple members that have some shared biochemical properties yet have unique biological roles. Although HIF members are the major substrates identified to date for the PHD members, a reasonable expectation is that other proteins whose activities are altered by hypoxia may also serve as PHD substrates. Indeed, the beta(2)-adrenergic receptor, a major adrenergic heterotrimeric guanine nucleotide-binding protein-coupled receptor in the heart, has been identified as a substrate for PHD3. PMID- 19903932 TI - Sex steroid hormones, hormonal contraception, and the immunobiology of human immunodeficiency virus-1 infection. AB - Worldwide, an increasing number of women use oral or injectable hormonal contraceptives. However, inadequate information is available to aid women and health care professionals in weighing the potential risks of hormonal contraceptive use in individuals living with HIV-1 or at high risk of infection. Numerous epidemiological studies and challenge studies in a rhesus macaque model suggest that progesterone-based contraceptives increase the risk of HIV-1 infection in humans and simian immunodeficiency virus (SIV) infection in macaques, accelerate disease progression, and increase viral shedding in the genital tract. However, because several other studies in humans have not observed any effect of exogenously administered progesterone on HIV-1 acquisition and disease progression, the issue continues to be a topic of intense research and ongoing discussion. In contrast to progesterone, systemic or intravaginal treatment with estrogen efficiently protects female rhesus macaques against the transmission of SIV, likely by enhancing the natural protective properties of the lower genital tract mucosal tissue. Although the molecular and cellular mechanisms underlying the effect of sex steroid hormones on HIV-1 and SIV acquisition and disease progression are not well understood, progesterone and estrogen are known to regulate a number of immune mechanisms that may exert an effect on retroviral infection. This review summarizes current knowledge of the effects of various types of sex steroid hormones on immune processes involved in the biology of HIV-1 infection. PMID- 19903937 TI - A new mechanism of phosphoregulation in signal transduction pathways. AB - Histidine protein kinases and serine, threonine, or tyrosine protein kinases play essential roles in signal transduction in prokaryotes and eukaryotes. A third type of protein kinase, an arginine protein kinase, has been identified. McsB of Bacillus subtilis phosphorylates the heat shock transcriptional regulator CtsR and can be regarded as the founding member of arginine protein kinases. PMID- 19903938 TI - A gluconeogenic tryst in the nucleus, with ER stress as the third wheel. AB - The transcriptional output of a cell reflects the sum of cooperative and competing interactions among hundreds of transcriptional regulators that are themselves regulated according to cellular conditions. In this way, disparate signaling cascades intersect at the level of gene expression; perturbation in one area of the cell will necessarily and fundamentally affect other areas as the cell strives to integrate information from multiple pathways. The consequences of such transcriptional cross-talk are highlighted by a newly discovered connection, through a common co-regulator, between stress in the endoplasmic reticulum and control of gluconeogenesis. These findings hint at the possible functions of stress pathways as regulators of basal cellular homeostasis. PMID- 19903939 TI - FBXO31: a new player in the ever-expanding DNA damage response orchestra. AB - The DNA damage response (DDR)-a central axis in the maintenance of genomic stability-has emerged as a complex signaling network that affects many aspects of cellular metabolism. A major arm of the DDR activates special checkpoints that temporarily arrest cell cycle progression while damage is being assessed and processed. Many DDR arms are driven by several parallel pathways acting in concert. Such is the case with the damage-induced G(1)/S checkpoint. A new pathway driving this checkpoint draws attention to the complexity of the DDR, which allows tight but fine-tuned control of the cellular response to threats to genomic integrity. PMID- 19903940 TI - Nutrition-minded cell cycle. AB - For decades, the fission yeast Schizosaccharomyces pombe has been used as an excellent model with which to explore how cellular growth is coordinated with the division cycle, a yet-unanswered question in biology. New studies in this organism show that TOR (target of rapamycin) kinase and stress-responsive MAPK (mitogen-activated protein kinase) form a signaling pathway that readjusts the timing of mitotic onset in response to poor nutrient conditions. Nutritional environment appears to be translated into graded activity of the protein kinases that influence the activation of Cdc2, a cyclin-dependent kinase driving cell cycle progression. PMID- 19903943 TI - Influence of different litter materials on cecal microbiota colonization in broiler chickens. AB - A chicken growth study was conducted to determine if litter type influenced gut microbiota and performance in broilers. Seven bedding materials were investigated and included soft and hardwood sawdust, softwood shavings, shredded paper, chopped straw, rice hulls, and reused softwood shavings. Microbial profiling was done to investigate changes in cecal bacterial communities associated with litter material and age. Cecal microbiota were investigated at 14 and 28 d of age (n = 12 birds/litter material). At both ages, the cecal microbiota of chickens raised on reused litter was significantly (P < 0.05) different from that of chickens raised on any of the other litter materials, except softwood shavings at d 28. Cecal microbiota was also significantly different between birds raised on shredded paper and rice hulls at both ages. Age had a significant influence on cecal microbiota composition regardless of litter material. Similarity in cecal microbial communities among birds raised on the same litter treatment was greater at 28 d of age (29 to 40%) than at 14 d of age (25 to 32%). Bird performance on the different litter materials was measured by feed conversion ratio, live weight, and feed intake. Significant (P < 0.05) differences were detected in live weight at 14 d of age and feed intake at 14 and 28 d of age among birds (n = 160/treatment) raised on some of the different litter materials. However, no significant (P > 0.05) differences were observed in feed conversion ratio among birds raised on any of the 7 different litter materials at either 14 or 28 d of age. The type of litter material can influence colonization and development of cecal microbiota in chickens. Litter-induced changes in the gut microbiota may be partially responsible for some of the significant differences observed in early rates of growth; therefore, litter choice may have an important role in poultry gut health particularly in the absence of in-feed antibiotics. PMID- 19903941 TI - H2S signals through protein S-sulfhydration. AB - Hydrogen sulfide (H2S), a messenger molecule generated by cystathionine gamma lyase, acts as a physiologic vasorelaxant. Mechanisms whereby H2S signals have been elusive. We now show that H2S physiologically modifies cysteines in a large number of proteins by S-sulfhydration. About 10 to 25% of many liver proteins, including actin, tubulin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), are sulfhydrated under physiological conditions. Sulfhydration augments GAPDH activity and enhances actin polymerization. Sulfhydration thus appears to be a physiologic posttranslational modification for proteins. PMID- 19903933 TI - The effects of opioids and opioid analogs on animal and human endocrine systems. AB - Opioid abuse has increased in the last decade, primarily as a result of increased access to prescription opioids. Physicians are also increasingly administering opioid analgesics for noncancer chronic pain. Thus, knowledge of the long-term consequences of opioid use/abuse has important implications for fully evaluating the clinical usefulness of opioid medications. Many studies have examined the effect of opioids on the endocrine system; however, a systematic review of the endocrine actions of opioids in both humans and animals has, to our knowledge, not been published since 1984. Thus, we reviewed the literature on the effect of opioids on the endocrine system. We included both acute and chronic effects of opioids, with the majority of the studies done on the acute effects although chronic effects are more physiologically relevant. In humans and laboratory animals, opioids generally increase GH and prolactin and decrease LH, testosterone, estradiol, and oxytocin. In humans, opioids increase TSH, whereas in rodents, TSH is decreased. In both rodents and humans, the reports of effects of opioids on arginine vasopressin and ACTH are conflicting. Opioids act preferentially at different receptor sites leading to stimulatory or inhibitory effects on hormone release. Increasing opioid abuse primarily leads to hypogonadism but may also affect the secretion of other pituitary hormones. The potential consequences of hypogonadism include decreased libido and erectile dysfunction in men, oligomenorrhea or amenorrhea in women, and bone loss or infertility in both sexes. Opioids may increase or decrease food intake, depending on the type of opioid and the duration of action. Additionally, opioids may act through the sympathetic nervous system to cause hyperglycemia and impaired insulin secretion. In this review, recent information regarding endocrine disorders among opioid abusers is presented. PMID- 19903944 TI - Effect of the density of conspecifics on runway social reinstatement behavior of male Japanese quail genetically selected for contrasting adrenocortical responsiveness to stress. AB - Runway tests are considered indicative of underlying sociality in birds and their ability to make social discriminations and establish interactions among conspecifics. Herein, social reinstatement behavior in male juvenile Japanese quail selected for a reduced (LS, low stress) or exaggerated (HS, high stress) adrenocortical response to brief mechanical restraint was evaluated. Individual males were given the choice to reinstate with either 2 (low density, LD) or 8 (high density, HD) unfamiliar randombred conspecifics placed in goal boxes at opposite ends of a two-choice runway (TCRW). Then, the same males were individually retested in a single goal box runway wherein they were exposed to a goal box containing either a LD or a HD of males. In the TCRW, a higher (P < 0.01) number of HS males started their ambulation toward the goal box containing the HD as opposed to LD of conspecifics. The HS males also spent more (P < 0.01) time in close proximity (within a 10-cm close zone; CZ) to the HD (218 s) rather than LD (57 s) of conspecifics. In contrast, the LS males did not differ in their initial direction of travel and they spent similar average amounts of times in the CZ of their stimulus LD (141 s) and HD (124 s) conspecifics. Similar to the TCRW results, in the single goal box runway, HS males spent more (P < 0.01) time in the CZ of HD rather than LD conspecifics, whereas LS quail spent similar amounts of time in the CZ of LD and HD males. Considering that runways are novel (and therefore frightening) environments, our findings suggest that HS quail may find better shelter (i.e., more comfort) in close proximity to a larger rather than smaller group of conspecifics, whereas LS birds find groups of varying conspecific density equally attractive. The results suggest that LS quail possess favorable social adaptive qualities because they appear to be better suited to cope with situations where the density of conspecifics is variable. PMID- 19903945 TI - Effect of the housing system on shedding and colonization of gut and internal organs of laying hens with Salmonella Enteritidis. AB - As a result of welfare considerations, conventional cage systems will be banned in the European Union from 2012 onward. Currently, there is limited information on the level of contamination with zoonotic pathogens related to the laying hen housing system. Therefore, 2 studies were designed to investigate the effect of the housing system on colonization of layers with Salmonella. In both studies, layers were housed in 3 different housing systems: a conventional cage system, a furnished cage, and an aviary. At 18 wk of age, all birds were orally inoculated with Salmonella Enteritidis. Shedding and colonization were measured at regular time points. The results did not show an increased risk for alternative housing systems compared with the conventional battery cage system. In contrast, in one study, a faster decline in shedding was noted for layers housed in the alternative cage systems in comparison with the conventional cage system. This study does not give indications that housing layers in alternative systems will yield a risk for increased Salmonella contamination. PMID- 19903946 TI - Quantitative determination of nutrient content in poultry manure by near infrared spectroscopy based on artificial neural networks. AB - Excessively applied manure contains a considerable amount of nutrient content such as nitrogen and phosphorus that could potentially pollute groundwater and soil. The present paper evaluated the use of nonlinear regression methods, such as artificial neural networks (ANN), for developing near infrared reflectance spectroscopy calibration models to predict nutrient content in poultry manure. Four representative nutrient ingredients (ammonia nitrogen, AN; total potassium, TK; total nitrogen, TN; total phosphorus, TP) in poultry manure were selected for evaluating ANN feasibility using 91 diverse samples in which three-fourths of the samples were used as a training set and one-fourth as a validation set. The performance of the ANN models was compared with the partial least squares (PLS) models. We found that the ANN models for all 4 nutrient contents consistently gave better predictions than PLS models. The ratios of prediction to deviation of 2.62 (AN), 1.51 (TK), 2.75 (TN), and 2.01 (TP) with the PLS models were improved to 3.02 (AN), 1.74 (TK), 3.41 (TN), and 2.71 (TP) with the corresponding ANN models. These findings demonstrated that the near infrared reflectance spectroscopy model based on the ANN method may be an appropriate tool to predict nutrient content in poultry manure. PMID- 19903947 TI - The influence of the lighting program on broiler activity and dust production. AB - Among the factors influencing dust production on the farms, animal activity is probably the least studied. Animal activity is strongly affected by circadian rhythms and it is altered by the rearing conditions, especially the management of light and the feeding strategy. In this study, a broiler flock was evaluated until 35 d of age with the general objective of studying the factors influencing the production of dust, particularly the effect of animal activity, as affected by the lighting program. In an experimental room, 2 daily dark periods of 6 and 4 h with 158 birds divided in 12 groups, reared on wood shavings, were evaluated. A TEOM analyzer was used to measure dust concentrations, whereas animal activity was determined by observation of birds on video tapes. Animal activity was defined by an activity index, which was determined by direct evaluation of birds on video tapes. The animal activity index was closely related to the lighting program (0.084 and 0.556 during dark and light periods, respectively) and was maximum at wk 4 of the rearing period. The dust concentrations during light periods were on average 4 times higher than during dark periods, with a maximum average concentration of 2.82 mg x m(-3) during the last week of the cycle. Particulate matter emission was on average 2.08 mg x animal(-1) x hour(-1). Dust concentration increased linearly with bird weight and daily variations in dust production were characterized. As a result, a direct cause-effect relationship between animal activity and dust concentration was obtained (r(2) = 0.89). PMID- 19903948 TI - Polymorphism of the ovocalyxin-32 gene and its association with egg production traits in the chicken. AB - We performed candidate gene analysis to identify SNP in the chicken ovocalyxin-32 (OCX-32) gene in the F(2) resource population, to develop a PCR-RFLP method for genotyping and to evaluate the associations of the gene polymorphism with egg production traits. The F(2) resource population-comprising 272 chickens-was obtained by crossing White Leghorn (WL) males and Rhode Island Red (RIR) females. They were measured for egg production traits and used for candidate gene analysis. Among parental individuals of the F(2) population, 2 novel nonsynonymous polymorphisms (c.267T>G and c.494A>C) and 1 known nonsynonymous polymorphism (c.381G>C) in the coding sequences of the chicken OCX-32 gene were detected. The PCR-RFLP method was used for screening the chickens of the F(2) population. In parental populations, genotype c.267T>G and c.494A>C were segregated within WL and RIR breeds, respectively, but genotype c.381G>C was breed-specific SNP between WL and RIR breeds. A total of 4 haplotypes were constructed based on the 3 SNP in parental populations, and there was no recombination between c.267T>G and c.494A>C. There was a significant association (P < 0.05) between the OCX-32 gene SNP and egg production traits, but there was no significant association between the haplotypes of the OCX-32 gene and egg production traits in the F(2) population. In the present study, there was the most significant association between c.381G>C of the OCX-32 gene and rate of egg production. The current study is the first step to confirm the relationship between OCX-32 gene polymorphisms and egg production traits. PMID- 19903949 TI - Evaluation of diversity between different Spanish chicken breeds, a tester line, and a White Leghorn population based on microsatellite markers. AB - The present study was conducted to evaluate the genetic variability and the genetic divergence of 13 Spanish chicken breeds, a tester line, and a White Leghorn population, using 24 microsatellite markers. A total of 150 alleles were detected across all population. The number of alleles by locus ranged from 2 to 13, with the mean value being 6.25. The mean polymorphic information content was 0.591, ranging from 0.847 to 0.172. The combined parentage exclusion probability of excluding 1 parent or 2 parents was 99 and 100%, respectively. The observed heterozygosity was lower than the expected heterozygosity for all loci, the mean values being 0.461 and 0.637. The observed and expected heterozygosity ranged from 0.003 to 0.735 and 0.181 to 0.863, respectively. Mean deficit of heterozygotes within populations (F(IS)) was 0.056 and mean fixation index of each population (F(ST)) was 0.244. The mean global deficit of heterozygotes across populations (F(IT)) was 0.286. A total of 15 private alleles in 10 microsatellites were observed, and in some populations, fixed alleles were found for 7 microsatellites. A total of 300 birds (83%) were properly assigned to the source population. The average observed heterozygosity for each population was 0.461, ranging from 0.328 (Quail Castellana) to 0.538 (Red Villafranquina), and the average expected heterozygosity was 0.488, ranging from 0.320 (Quail Castellana) to 0.550 (White-Faced Spanish). All of the Spanish breeds except the Quail Castellana were more polymorphic than the White Leghorn population. The mean value of the deviation of heterozygote number was 0.052. Nei's genetic distance showed a range from 0.109 (between White-Faced Spanish and Black Menorca) to 0.437 (between Buff Prat and White Leghorn). A phylogenetic tree constructed by the neighbor-joining method, based on Nei's genetic distance, showed a clear separation between the White Leghorn and the remaining breeds. The results indicate that the panel of microsatellite markers was useful in studying the genetic diversity of chicken breeds. PMID- 19903950 TI - Multiplex polymerase chain reaction for the detection and differentiation of avian influenza viruses and other poultry respiratory pathogens. AB - A multiplex reverse transcription-PCR (mRT-PCR) was developed and standardized for the detection of type A influenza viruses, avian influenza virus (AIV) subtype H7, H9, and H5 hemagglutinin gene with simultaneous detection of 3 other poultry respiratory pathogens, Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and infectious laryngotracheitis virus (ILTV). Seven sets of specific oligonucleotide primers were used in this study for the M gene of AIV and hemagglutinin gene of subtypes H7, H9, and H5 of AIV. Three sets of other specific oligonucleotide primers were used for the detection of avian respiratory pathogens other than AIV. The mRT-PCR DNA products were visualized by agarose gel electrophoresis and consisted of DNA fragments of 1,023 bp for M gene of AIV, 149 bp for IBV, 320 bp for NDV, and 647 bp for ILTV. The second set of primers used for m-RT-PCR of H7N3, H9N2, and H5N1 provided DNA products of 300 bp for H7, 456 bp for H5, and 808 bp for H9. The mRT-PCR products for the third format consisted of DNA fragments of 149 bp for IBV, 320 bp for NDV, 647 bp for ILTV, 300 bp for H7, 456 bp for H5, and 808 bp for H9. The sensitivity and specificity of mRT-PCR was determined and the test was found to be sensitive and specific for the detection of AIV and other poultry respiratory pathogens. In this present study, multiplex PCR technique has been developed to simultaneously detect and differentiate the 3 most important subtypes of AIV along with the 3 most common avian respiratory pathogens prevalent in poultry in Pakistan. Therefore, a mRT PCR that can rapidly differentiate between these pathogens will be very important for the control of disease transmission in poultry and in humans, along with the identification of 3 of the most common respiratory pathogens often seen as mixed infections in poultry, and hence economic losses will be reduced in poultry. PMID- 19903951 TI - Immunomodulation in gut-associated lymphoid tissue of neonatal chicks by immunobiotic diets. AB - Developmental changes in immunocompetent cells of the gut during the first week posthatch were determined in broiler chicks fed immunobiotic lactic acid bacteria in the form of Lactobacillus jensenii TL2937-, Lactobacillus gasseri JCM1131(T)-, Lactobacillus delbrueckii ssp. bulgaricus NIAIB6-, or L. gasseri TL2919 supplemented diets. The relative weights of spleen and bursa of Fabricius in chicks fed the immunobiotic diets were slightly higher than the control valued at 1 and 3 d of age, with the exception of spleen weight in the L. gasseri JCM1131(T) at 3 d of age, the bursa of Fabricius weight in the L. gasseri JCM1131(T) at 1 and 3 d of age, and bursa of Fabricius weight in the L. gasseri TL2919 group at 1 d of age. There were no significant differences in body and liver weights among the treatments. When chicks were fed the L. jensenii TL2937- or L. gasseri TL2919-supplemented diets, expression of T cell-related mRNA [cluster of differentiation 3 (CD3), interleukin-2 (IL-2), and interferon-gamma (IFN-gamma)] in the foregut was significantly higher than that of control chicks at 3 or 7 d of age. Expression levels of toll-like receptor (TLR) mRNA tended to increase in the foregut of chicks fed the immunobiotic diets, except for the L. delbrueckii ssp. bulgaricus NIAIB6, compared with expression levels in control chicks. The Bu-1 mRNA expression levels in the bursa of Fabricius were not affected by the supplementations with immunobiotic lactic acid bacteria. These results show that immunobiotics, particularly L. gasseri TL2919, might be useful as immunomodulators to stimulate the gut-associated immune system in neonatal chicks, and thereby protect them from disease without decreasing growth performance as a possible substitution of antibiotics. PMID- 19903952 TI - Effects of methyltestosterone on immunity against Salmonella Pullorum in dwarf chicks. AB - This study was conducted to determine effects of methyltestosterone on innate immunity and adaptive immunity against Salmonella Pullorum in dwarf chicks. In vivo experiment, comparisons of pathological sections, viable counts of bacteria, specific antibody levels, and subsets of T lymphocytes were set forth between chicks with or without 10(-7) M methyltestosterone treatment (2 d of age through 21 d of age) and challenged with 5 x 10(8) virulent Salmonella Pullorum (7 d of age), and in vitro experiment, phagocytic and killing abilities, reactive oxygen intermediate production, and reactive nitrogen intermediate production of monocytes-macrophages treated with high (10(-8) M/10(6) cell) or physiological (10(-14) M/10(6) cell) concentration of methyltestosterone were examined after Salmonella Pullorum infection. The results showed that (1) in vivo, administration of methyltestosterone enhanced susceptibility to Salmonella Pullorum infection and depressed cellular immunity against Salmonella Pullorum, whereas it had no effect on humoral immunity in dwarf chicks; (2) in vitro, at high concentration, methyltestosterone reduced (P < 0.05) monocytes-macrophages mediated reactive oxygen intermediate-dependent killing of Salmonella Pullorum, whereas low concentration of methyltestosterone enhanced (P < 0.05) reactive oxygen intermediate-dependent killing of Salmonella Pullorum in male dwarf chicks but not in females; and (3) although challenged with Salmonella Pullorum, phagocytic ability and monocytes-macrophages mediated reactive nitrogen intermediate-dependent killing were not affected by methyltestosterone in vitro. The results indicated that methyltestosterone affected the immune response to Salmonella Pullorum in dwarf chicks by changing monocytes-macrophages mediated reactive oxygen intermediate-dependent killing and cellular immunity, and the effects were dose-dependent; furthermore, the former 2 pathways played important roles in preventing Salmonella Pullorum infection in dwarf chicks, although the mechanism needs further study. PMID- 19903953 TI - Effects of cold stress on the messenger ribonucleic acid levels of peroxisome proliferator-activated receptor-{gamma} in spleen, thymus, and bursa of Fabricius of chickens. AB - This study was to investigate the expression trait of the peroxisome proliferator activated receptor-gamma (PPAR-gamma) gene and the effect of cold stress on the mRNA levels of PPAR-gamma in spleen, thymus, and bursa of Fabricius of chickens. Eighty-four 1-d-old male chickens were randomly allocated to 12 groups (7 chickens per group). There was 1 control group and 5 treatment groups for acute cold stress and 3 control groups and 3 treatment groups for chronic cold stress. Chickens were maintained in our animal facility, kept under a 16L:8D cycle and temperature (30 +/- 2 degrees C), and given free access to standard chow and water. The cold stress was initiated when the birds were 15 d of age, with the duration of the acute cold stress being 1, 3, 6, 12, and 24 h, and the chronic cold stress was 5, 10, and 20 d, respectively. Cold stress temperature was 12 +/- 1 degrees C. Spleen, thymus, and bursa of Fabricius were collected for the assessment of the mRNA levels by real-time PCR after stress termination. The results showed that the PPAR-gamma gene is expressed in spleen, thymus, and bursa of Fabricius, and its expression level is different in different tissues and at different ages. Acute cold stress significantly decreased (P < 0.05) the mRNA levels of the PPAR-gamma gene of spleen and thymus in all treatment groups and significantly increased (P < 0.05) the mRNA levels of the PPAR-gamma gene of bursa of Fabricius in all treatment groups. Compared with the corresponding control groups, chronic cold stress resulted in a significant increase (P < 0.05) of the mRNA levels of the PPAR-gamma gene in spleen and a significant decrease (P < 0.05) of the mRNA levels of the PPAR-gamma gene in thymus and bursa of Fabricius. The results indicate that the PPAR-gamma gene is expressed in all 3 immune organs and has different expression traits. The magnitude and direction of change in PPAR-gamma gene expression differs with the type of cold stress applied and also varies by tissue. PMID- 19903954 TI - Long-term effects of feeding flaxseeds on hepatic lipid characteristics and histopathology of laying hens. AB - The long-term effects of dietary flaxseed and tocopherols on hepatic and blood plasma lipids, fatty acids, tocopherols, hepatic TBA reactive substances, and histopathology of Brown Leghorn hens were investigated. Thirty-two-week-old ISA Brown Leghorn hens (n = 120) were kept in cages and were fed 1 of the 3 corn soybean meal-based diets, a control diet (no flax) or a 10% flax diet with or without 100 IU of tocopherols, until the hens were 64 wk of age. Feeding diets with 10% flaxseed reduced hepatic and plasma fat content, hepatic triglycerides, total number of fat vacuoles, and number of cells with 75% or higher lipid infiltration in hepatocytes (P < 0.05). Addition of tocopherols to the 10% flax diet increased hepatic and plasma tocopherol content. A significant reduction in hepatic TBA reactive substances was observed in the hens supplemented with the basal diet with 10% flax and 100 IU of tocopherols (P < 0.05). Feeding diets containing flaxseed resulted in an increase in the content of alpha-linolenic (18: n-3) and docosahexaenoic acids (22:6n-3), with a concomitant reduction in monounsaturated and saturated fatty acids in the hen liver and plasma. Overall, long-term feeding of hens with flax led to a reduction in liver and plasma lipids and reduced hepatocellular infiltration. Inclusion of tocopherols may be needed to reduce lipid oxidation products in the liver of flax-fed hens. However, tocopherol supplementation had no effect on hepatocellular lipid infiltration or liver total lipid or triglyceride content. PMID- 19903955 TI - Effect of dietary fiber and fat on performance and digestive traits of broilers from one to twenty-one days of age. AB - The influence of fiber source and dietary fat level on digestive traits and productive performance was studied in broilers from 1 to 21 d of age. There were 6 treatments arranged factorially with 3 sources of fiber (none; 3% oat hulls, OH; and 3% sugar beet pulp, SBP) and 2 fat sources (5% soybean oil, SO; and 5% yellow grease, YG). Each treatment was replicated 6 times and the experimental unit was a cage with 18 broilers. Fiber inclusion improved BW gain (P < or = 0.05) and feed:gain ratio (P < or = 0.001) and increased total tract apparent retention (TTAR) of all nutrients measured (P < or = 0.001). The increases observed in TTAR of nitrogen and ether extract and on AME(n) of the diet were more pronounced with OH than with SBP. The increases in nutrient digestibility with OH inclusion were higher at excreta than at ileal level and in fact, SBP inclusion reduced the apparent ileal digestibility of most nutrients. The relative weight (%) of the gizzard was increased (P < or = 0.001) and the pH of its contents was reduced (P < or = 0.001) when additional fiber was included in the diet. The TTAR of nutrients was higher for the SO than for the YG diets (P < or = 0.001). Also, the increases in ether extract digestibility (P < or = 0.05) and AME(n) (P < or = 0.05) of the diet with fiber inclusion were more pronounced with the YG than with the SO. Therefore, the inclusion of moderate amounts of fiber in the diet might improve performance and nutrient digestibility in young chicks, especially when saturated fats are used. PMID- 19903956 TI - Live performance, carcass composition, and blood metabolite responses to dietary nutrient density in two distinct broiler breeds of male chickens. AB - A study was conducted to evaluate the effects of varying nutrient density with constant ME:CP ratio on growing performance, carcass characteristics, and blood responses in 2 distinct broiler breeds of male chickens (Arbor Acres, a commercial line, and Beijing-You, a Chinese nonimproved line). Experimental diets were formulated with high, medium, or low nutrient densities for 3 growing phases. Starter diets (1 to 21 d) contained 23, 21, and 19% CP with 3,059, 2,793, and 2,527 kcal/kg of ME; grower diets (22 to 35 d) contained 21, 19, and 17% CP with 3,150, 2,850, and 2,550 kcal/kg of ME; and finisher diets (36 to 42 d for Arbor Acres and 36 to 91 d for Beijing-You) had 19, 17, and 15% CP with 3,230, 2,890, and 2,550 kcal/kg of ME. Male hatchlings (216 of each breed) were randomly assigned to 6 replicates of 12 birds in each treatment. Arbor Acres broilers had better (P < 0.001) BW gain, feed conversion ratio (FCR), and carcass yield, but had greater (P < 0.001) abdominal and carcass fat deposition. In both breeds, the higher nutrient density increased (P < 0.05) BW gain, protein efficiency ratio, and energy efficiency ratio while decreasing (P < 0.05) feed intake and FCR. The breed differences were increased for FCR, protein efficiency ratio, and energy efficiency ratio in the starter period and decreased for carcass chemical composition, respectively, by higher nutrient density. These findings indicate that 1) genetic improvement has a significant effect on broiler responses to dietary nutrient density, 2) performance differences between breeds are lessened with diets of low nutrient density, 3) carcass quality differences are less when birds were fed diets of high nutrient density, 4) carcass composition is hardly modified by nutrient density and both breeds exhibit similar metabolite responses to dietary concentrations, and 5) optimal diets are deduced for these breeds for the 3 growing phases. PMID- 19903957 TI - Regulation of adipose triglyceride lipase by fasting and refeeding in avian species. AB - Lipolysis in fat tissue is a process that is not fully understood. Increasing knowledge of the process could allow for increased feed efficiency and reduced fat content, which would lower feeding costs for poultry production. Adipose triglyceride lipase (ATGL) is an adipose-specific enzyme that cleaves at the Sn-1 position of triglycerides, releasing nonesterified fatty acids (NEFA) into the bloodstream. Adipose triglyceride lipase has recently been cloned in avian species. For further understanding of how ATGL responds to environmental stimuli, we fasted 21-d-old Ross 308 broiler chickens for 24 h. Adipose and liver tissues were collected before the fasting period and at its conclusion, as well as 4, 8, 12, and 24 h after being refed. Blood samples were also collected at these time points. Additionally, tissue samples were collected from 30 quails subjected to the same fasting period, with refeeding time points of 2, 4, and 8 h. Adipose triglyceride lipase in tissue samples was analyzed via Western blot and quantitative real-time PCR. Protein and RNA levels of ATGL were high in the birds after the fasting period. Ribonucleic acid levels quickly returned to control levels following refeeding. Protein levels, however, remained high in the chicken throughout the 4- and 8-h refeeding time points. For the quail samples, ATGL protein returned to normal levels at 8 h. To relate the release of NEFA into the blood with ATGL expression, plasma analysis was done. Nonesterified fatty acids were significantly higher after the fasting period than the control and returned to control levels by 4 h after refeeding. The quick return of the RNA to control levels suggests that ATGL production was stimulated during the fasting period but inhibited once food was reintroduced. The immediately lowered NEFA levels suggest that the residual high amounts of ATGL protein shown by Western blot were no longer functioning. This suggests the existence of a mechanism to inactivate the active form of ATGL, possibly through posttranslational modification of the protein. PMID- 19903958 TI - Standardized ileal amino acid digestibility in wheat distillers dried grains with solubles for broilers. AB - A study was conducted to estimate the ileal digestibility of amino acids (AA) in 5 different samples of wheat distillers dried grains with solubles (DDGS) fed to broilers. Two hundred sixteen male Ross broiler chicks were fed a commercial starter diet from d 1 to 15 of age followed by the test diets from d 15 to 21. The 5 test diets consisted of a DDGS sample or wheat as the sole source of AA, dextrose, minerals, and vitamins. Chromic oxide (0.3%) was included in all diets as a digestibility marker. Each test diet was randomly assigned to 6 replicate cages, each with 6 birds. On d 21, birds were killed to sample ileal digesta for determining the apparent (AID) and standardized (SID) ileal AA digestibilities. The SID values were calculated using ileal endogenous AA losses previously determined in our laboratory. Among the indispensable AA in wheat DDGS, the lowest and highest AID average values were observed for Lys (35.6%) and Phe (79.2%), respectively. The most variable AID estimates of wheat DDGS samples were observed for Lys (24.4 to 45.7%), Thr (48.2 to 60.9%), and His (57.4 to 69.1%) as indispensable and Asp (32.5 to 50.9%), Gly (49.6 to 63.1%), and Ala (53.6 to 66.8%) as dispensable AA, respectively. Apparent ileal digestibility estimates of the wheat sample for Lys, Thr, His, Gly, and Ala were 77.5, 74, 83.6, 79.3, and 78.9%, respectively. All AA digestibility estimates for both AID and SID determined in wheat were higher than in wheat DDGS samples (P < 0.05). Considering both AID and SID coefficients of wheat DDGS samples, Lys was the least digestible AA, averaging 35.6 and 40.0%, respectively. Using SID values in practical diet formulation can increase accuracy, prevent overformulation of diets, and reduce cost of safety margins. PMID- 19903959 TI - Prehatch intestinal maturation of turkey embryos demonstrated through gene expression patterns. AB - Some of the challenges faced by neonatal turkeys include weakness, reduced feed intake, impaired growth, susceptibility to disease, and mortality. These symptoms may be due to depleted energy reserves after hatch and an immature digestive system unable to replenish energy reserves from consumed feed. To better understand enteric development in turkeys just before hatch, a new method was used to identify the patterns of intestinal gene expression by utilizing a focused microarray. The duodenums of 24 turkey embryos were sampled on embryonic day (E)20, E24, E26, and hatch (E28). The RNA populations of 96 chosen genes were measured at each time point, from which 81 significantly changed (P < 0.01). These genes were clustered by gene expression pattern similarity into 4 groups. The expression pattern of hormone receptors revealed that intestinal tissues may be less responsive to growth hormone, insulin, glucagon, and triiodothyronine during the last 48 h before hatch, when developmental emphasis switches from cell proliferation to functional maturation. Based on gene expression patterns, we concluded that at hatch, poults should have the capacity to 1) digest disaccharides but not oligopeptides, due to increased expression of sucrase isomaltase but decreased expression of aminopeptidases and 2) absorb monosaccharides and small peptides due to high expression of sodium-glucose cotransporter-4 and peptide transporter-1. PMID- 19903960 TI - Comparison of a modern broiler line and a heritage line unselected since the 1950s. AB - Selecting chicken for improved meat production has altered the relative growth of organs in modern broiler lines compared with heritage lines. In this study, we compared the growth and feed efficiency of a heritage line, UIUC, with a modern production line, Ross 708, for 5 wk posthatch. During this period, the BW and feed efficiency of the modern strain was higher than that of the heritage line, indicating that the Ross 708 birds were more efficient than the UIUC birds at converting feed to body mass. The relative growth of the breast, heart, liver, and intestine were also compared during these 5 wk. The breast muscle of the heritage line constituted 9% of the total body mass at 5 wk, whereas in the modern line, the breast muscle was 18% of the total mass of the bird. In contrast, the relative size of the heart decreased after d 14 in the modern line, suggesting that selection for increased breast muscle has translated into relatively less weight of the heart muscle. The liver matured earlier in modern lines, possibly improving nutrient utilization as the birds shift from lipid- to carbohydrate-rich feed. Finally, jejunal and ileal sections of the intestine were 20% longer in the modern line, perhaps allowing for increased nutrient absorption. PMID- 19903961 TI - Effects of turmeric (Curcuma longa) on the expression of hepatic genes associated with biotransformation, antioxidant, and immune systems in broiler chicks fed aflatoxin. AB - The objective of the present study was to evaluate the efficacy of curcumin, an antioxidant found in turmeric (Curcuma longa) powder (TMP), to ameliorate changes in gene expression in the livers of broiler chicks fed aflatoxin B(1) (AFB(1)). Four pen replicates of 5 chicks each were assigned to each of 4 dietary treatments, which included the following: A) basal diet containing no AFB(1) or TMP (control), B) basal diet supplemented with TMP (0.5%) that supplied 74 mg/kg of curcumin, C) basal diet supplemented with 1.0 mg of AFB(1)/kg of diet, and D) basal diet supplemented with TMP that supplied 74 mg/kg of curcumin and 1.0 mg of AFB(1)/kg of diet. Aflatoxin reduced (P < 0.05) feed intake and BW gain and increased (P < 0.05) relative liver weight. Addition of TMP to the AFB(1) diet ameliorated (P < 0.05) the negative effects of AFB(1) on growth performance and liver weight. At the end of the 3-wk treatment period, livers were collected (6 per treatment) to evaluate changes in the expression of genes involved in antioxidant function [catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S-transferase (GST)], biotransformation [epoxide hydrolase (EH), cytochrome P450 1A1 and 2H1 (CYP1A1 and CYP2H1)], and the immune system [interleukins 6 and 2 (IL-6 and IL-2)]. Changes in gene expression were determined using the quantitative real-time PCR technique. There was no statistical difference in gene expression among the 4 treatment groups for CAT and IL-2 genes. Decreased expression of SOD, GST, and EH genes due to AFB(1) was alleviated by inclusion of TMP in the diet. Increased expression of IL-6, CYP1A1 and CYP2H1 genes due to AFB(1) was also alleviated by TMP. The current study demonstrates partial protective effects of TMP on changes in expression of antioxidant, biotransformation, and immune system genes in livers of chicks fed AFB(1). Practical application of the research is supplementation of TMP in diets to prevent or reduce the effects of aflatoxin in chicks fed aflatoxin contaminated diets. PMID- 19903962 TI - Liver X receptor alpha regulates fatty acid synthase expression in chicken. AB - Liver X receptor alpha (LXRalpha), also referred to as nuclear receptor subfamily 1, group H, member 3 is a member of the nuclear hormone receptor superfamily, and has recently been shown to act as a master transcription factor governing hepatic lipogenesis in mammals. Liver X receptor alpha directly regulates both the expression of other lipogenic transcription factors and the expression of lipogenic enzymes, thereby enhancing hepatic fatty acid synthesis (FASN). In birds, like in humans, fatty acid synthesis primarily occurs in the liver. Whether LXRalpha is involved in hepatic regulation of lipogenic genes remained to be investigated in this species. Here we show that fatty acid synthase and the expression of other lipogenic genes (sterol regulatory element binding protein 1 and steroyl coenzyme A desaturase 1) are induced in chicken hepatoma cells in response to a pharmacological liver X receptor agonist, T0901317. A detailed analysis of the chicken FASN promoter revealed a functional liver X response element. These data define the chicken FASN gene as a direct target of LXRalpha and further expand the role of LXRalpha as a regulator of lipid metabolism in this species. PMID- 19903963 TI - A possible role of chorion protease in shell membrane degradation during development of quail embryos. AB - In the eggs of the quail Coturnix japonica, the limiting membrane demarcates the shell membrane at the interface with the albumen and decreases in width during the hatching process. This study was done to identify agents that affect the width of this limiting membrane. Zymography tests on extracts from extraembryonic tissues, yolk sacs, or chorioallantoic membranes, or all three, showed proteolytic activities during d 4 to 10 of incubation. Localization experiments on these activities, performed on d 5 eggs, indicated that they were located in an avascular chorion. Electron microscopic analysis showed there were secretory cells specifically located in the avascular chorion. After partial purification of d 5 avascular chorion extracts using QA52 and Sephadex G-200 column chromatography, the proteolytic activity of 20 kDa was isolated. The protease showed a high level of activity toward succinyl-Gly-Pro-Leu-Gly-Pro-4 methylcoumaryl-7-amide. It had an optimal pH of 9 and digested the limiting membrane. These enzymatic activities were inhibited moderately by EDTA and strongly by leupeptin and aprotinin. It was concluded that it is the 20-kDa protease, showing collagenase-like activity produced by the avascular chorion, that affects the limiting membrane. PMID- 19903964 TI - Densitometric and biochemical values of broiler tibias at different ages. AB - The objective of this experiment was to determine the normal values of bone radiographic density (BRD) by using the optical densitometry in radiographic images and the biochemical values represented by serum calcium, ash percentage, and minerals (calcium, phosphorus, and magnesium) from tibia ash of Cobb broilers at 8, 22, and 43 d of age. A total of 14 broilers were used for densitometric analysis, and 15 were used for biochemical dosages. The BRD values increased (P < 0.05) with age and in all tibia regions (proximal epiphysis, diaphysis, and distal epiphysis), concluding that growth was a determinative factor for bone performance, demanding a higher BRD during broiler development. Tibia proximal epiphysis presented higher BRD values in relation to the other bone regions (P < 0.05), as a result of a possible biomechanical adaptation to ligaments and tension of the muscle tendons at this region, allowing the support of the muscle mass increase. The serum calcium values were kept constant, as a result of the appropriate nutritional levels of the diet that supported the animal homeostasis. The bone ash and mineral percentage increased (P < 0.05) at 22 d of age, due to the higher mineral requirement in this age. The correlation between bone densitometry and the invasive techniques showed that the bone densitometry can substitute the determination of mineral percentage in the ash. This experiment presented normal values of the noninvasive and invasive methods more used in aviculture, allowing us to compare, subsequently, pathological and physiological values or results of broilers fed with different diets. PMID- 19903965 TI - Influence of prestorage incubation on embryonic development, hatchability, and chick quality. AB - Egg storage longer than 7 d is associated with a delay in hatch time and a decline in hatchability and chick quality. Prestorage incubation is suggested as a method to reduce the negative effects of prolonged storage times by altering the developmental stage of the embryo, but earlier research has shown that prestorage incubation can both be detrimental and beneficial for hatchability. The reason for these ambiguous results is not clear and the effect of prestorage incubation on chick quality is not studied extensively. The objective of this study was to investigate changes in developmental stage of embryos during prestorage incubation and the effect of prestorage incubation on hatchability and chick quality. Two experiments were conducted. In experiment I, eggs were stored for 3, 5, 8, or 12 d. In experiment II, eggs were stored for 5 or 11 d. Half of the eggs was stored immediately at 16 to 18 degrees C and the other half was exposed to prestorage incubation for 6 h in experiment I and for 4.5 h in experiment II. According to the classification table of Eyal-Giladi and Kochav (EG), embryonic development was advanced by prestorage incubation from developmental stage EG11.67 to developmental stage EG13.26 in experiment I (P = 0.02) and from developmental stage EG9.22 to developmental stage EG12.63 in experiment II (P < 0.0001). In experiment I, prestorage incubation reduced hatchability of set eggs from 59.3 to 51.5% when storage time was 12 d but did not reduce hatchability when storage time was 3, 5, or 8 d (interaction P = 0.02). Prestorage incubation increased chick length (P = 0.004). In experiment II, prestorage incubation increased hatchability of fertile eggs from 80.6 to 85.9% when storage time was 11 d but did not increase hatchability when storage time was 5 d (interaction P = 0.0009). Prestorage incubation increased percentage of second grade chicks (P = 0.0007). It seems that storage time, embryonic development at egg collection, and prestorage incubation duration determine the effect of prestorage incubation on hatchability and chick quality. PMID- 19903966 TI - Use of the hypo-osmotic swelling test and aniline blue staining to improve the evaluation of seasonal sperm variation in native Spanish free-range poultry. AB - The season may affect the values of fresh semen variables and therefore influence the success of cryopreservation. The aim of this study was to improve the evaluation of seasonal changes in semen quality in Spanish Black Castellana roosters maintained under natural environmental conditions. Semen was collected from 11 Black Castellana roosters (housed under natural photoperiod and temperature conditions) by massage twice every month for 12 mo. In addition to determining ejaculate volume, sperm concentration, and sperm motility (the classic sperm variables), we used the hypo-osmotic swelling test to examine the membrane integrity of the spermatozoa. Further, morphological abnormalities and acrosome integrity were assessed via aniline blue staining. Semen volume (P < 0.05), sperm concentration (P < 0.01), and the percentage of spermatozoa with an intact acrosome (P < 0.01) were significantly affected by the season of the year. The annual profile of the percentage of spermatozoa showing acrosome integrity followed a trend roughly parallel to annual variations in temperature (Spearman rank correlation = 0.77, P < 0.01). According to the hypo-osmotic swelling test, membrane integrity fell in July (P < 0.05 compared with all other months), the month of highest temperatures. Aniline blue staining and the hypo-osmotic swelling test provide an easy and useful means of evaluating sperm abnormalities, including acrosome morphology and membrane integrity, and could be easily introduced into routine avian semen quality assessments. The results show that high semen quality is associated with long day photoperiods. Extreme heat or cold appear to exert a negative influence on sperm quality. PMID- 19903967 TI - The crystal polymorphism of calcium carbonate is determined by the matrix structure in quail eggs. AB - Two calcified structures, the eggshell and sperm-associated body (SB), are present in the eggs of the Japanese quail, Coturnix japonica. X-ray diffractometry showed that calcium carbonates take the form of calcite in the eggshell and aragonite in the SB. The aim of the present study was to identify the factors that determine the morphology of calcium carbonate crystals. The matrix of EDTA-treated eggshell was a meshwork of vesicles, 200 to 500 nm in diameter, connected by fine fibers or fibrous sheets. The matrix of SB cortex was a radiation of rod-shaped projections approximately 130 nm in width. In vitro crystal formation was achieved by adding dissociated matrix substances to test solutions. When eggshell matrix material was added, formation of calcite crystals, which had many vesicular holes on their surface, was observed. When SB matrix material dissociated by sonication was added, rhombohedral calcite crystals formed at protein concentrations of 100 microg/mL or lower, and elongated and bundled crystals formed at concentrations of 150 microg/mL or higher. When SB matrix material dissociated by pipetting was added, aragonite crystals formed. These observations indicate that the matrix structure is the principal factor in determining the crystal polymorphism of calcium carbonate. PMID- 19903968 TI - Thermal manipulations of broiler embryos--the effect on thermoregulation and development during embryogenesis. AB - This study aimed to elucidate the effects of thermal manipulations (TM) of broiler embryos, during the development of the thyroid and adrenal axis, on embryo development and metabolism. Cobb eggs were divided into 3 treatments: control, 24H-continuous TM at 39.5 degrees C and 65% RH from embryonic day 7 to 16 inclusive, and 12H-intermittent TM for 12 h/d in the same period. Only the 24H treatment negatively affected embryo growth and development, with lower relative weights of embryo, liver, and pipping muscle. During TM, eggshell temperature, heart rate, and oxygen consumption were elevated as embryos were in their ectothermic phase, but from the end of the TM until hatch, these parameters were significantly lower in both treatments than in the control. Moreover, plasma concentrations of the thyroid hormones were significantly lower in the 2 treatments during and after TM, until hatch. Plasma corticosterone concentration of the TM-treated embryos was significantly lower after the TM but significantly higher at hatch. It was concluded that TM during the development of the thyroid and adrenal axis lowered their functional set point, thus lowering metabolic rate during embryogenesis and at hatch. PMID- 19903969 TI - Highly efficient dissociation of oxygen from hemoglobin in Tibetan chicken embryos compared with lowland chicken embryos incubated in hypoxia. AB - Oxygen is one of the critical determinants for normal embryonic and fetal development. In avian embryos, lack of oxygen will lead to high fetal mortality, heteroplasia, and cardiovascular dysfunction. Tibetan chicken is a breed native to Tibet that could survive and keep higher hatchability regardless of negative effects of hypoxia. Generally, adaptive animals in high altitudes are characterized by higher hemoglobin concentrations and oxygen affinity. In the present study, the capacity of oxygen supply in late chick embryo (including d 17, 19, and 21) was compared between Tibetan chicken and a lowland breed, Dwarf White chicken, by determining the hemoglobin concentrations and oxygen equilibrium curves in both hypoxic (13% O(2)) and normoxic (21% O(2)) conditions. The results showed that a higher level of hemoglobin concentration was induced by hypoxia in Tibetan chicken embryos, and the hemoglobin could perform with better cooperativity and deliver oxygen to tissues more easily. Further investigation revealed that the carbonic anhydrase II mRNA in red blood cells of Tibetan chicken was increasingly induced to a higher level in hypoxia than that of the lowland breed. These results suggested that the stronger capacity of oxygen dissociation was an important characteristic of Tibetan chicken embryo to survive in hypoxia and the upregulating mode of carbonic anhydrase II mRNA might assist this dissociation. Therefore, for avian at high altitudes, the efficient dissociation of oxygen might reveal another aspect associated with the hypoxia adaptability. PMID- 19903970 TI - Salmonella serotype distribution in the Dutch broiler supply chain. AB - Salmonella serotype distribution can give insight in contamination routes and persistence along a production chain. Therefore, it is important to determine not only Salmonella prevalence but also to specify the serotypes involved at the different stages of the supply chain. For this purpose, data from a national monitoring program in the Netherlands were used to estimate the serotype distribution and to determine whether this distribution differs for the available sampling points in the broiler supply chain. Data covered the period from 2002 to 2005, all slaughterhouses (n = 22), and the following 6 sampling points: departure from hatchery, arrival at the farm, departure from the farm, arrival at the slaughterhouse, departure from the slaughterhouse, and end of processing. Furthermore, retail data for 2005 were used for comparison with slaughterhouse data. The following serotypes were followed throughout the chain: Salmonella Enteritidis, Salmonella Typhimurium, Salmonella Paratyphi B var. Java (Salmonella Java), Salmonella Infantis, Salmonella Virchow, and Salmonella Mbandaka. Results showed that serotype distribution varied significantly throughout the supply chain (P < 0.05). Main differences were found at the farm and at the slaughterhouse (within one stage), and least differences were found between departure from one stage and arrival at the next stage. The most prominent result was the increase of Salmonella Java at farm level. This serotype remained the most prominent pathogen throughout the broiler supply chain up to the retail phase. PMID- 19903971 TI - Buying higher welfare poultry products? Profiling Flemish consumers who do and do not. AB - A substantial number of studies has already investigated differences within the consumer market with regard to attitudes and perceptions in relation to farm animal welfare. Likewise, several studies focused on the gap that exists between positive attitudes and reported consumption or purchase intentions for sustainable food products in general and higher welfare products more specific, and on the factors influencing this attitude-behavior gap. Little or no studies, however, have started from reported pro-welfare behavior to distinguish between consumer groups and to explore the motivations of the respective behavior. With this study, we aim to group consumers according to their reported buying frequency of higher welfare eggs and higher welfare chicken meat. Similarities and dissimilarities between these groups are mapped in terms of individual characteristics, product attribute importance, perceived consumer effectiveness, perception of higher welfare products, and attitude toward a welfare label. The research methodology applied was a quantitative study with cross-sectional consumer survey data collected in Flanders in spring 2007 (n = 469). Pro-welfare behavior was unevenly distributed across different consumer segments, despite a general interest and concern for bird welfare. A consistent choice for standard (no welfare premium) poultry products was related to strong perceived price and availability barriers, to a low importance attached to ethical issues as product attributes, and to a low perceived consumer effectiveness. A consistent choice for products with higher welfare standards to the contrast associated with a high importance attached to ethical issues; a low effect of price and availability perception; a strong association of higher welfare products with product attributes like health, taste, and quality; and a high perceived consumer effectiveness. The identification of market segments with common characteristics is essential for positioning higher welfare products and developing effective communication strategies. Finally, a welfare label emerged as an appropriate communication vehicle for consumers who engage in pro-welfare behavior and who experienced the label as a solution to lower the search costs for higher welfare products. PMID- 19903972 TI - The effects of oviposition time on egg weight loss during storage and incubation, fertility, and hatchability of broiler hatching eggs. AB - An experiment was conducted to determine the effect of time of oviposition, generally representing different positions in the normal egg laying sequence, on egg weight loss during storage and incubation, and on fertility and fertile hatchability of eggs from mid-lay (42 wk) and old (67 wk) broiler breeders. A total of 1,800 eggs (900 eggs per flock age) were collected during 10 consecutive days between 0830 and 1830 h each day. The eggs were individually marked, weighed, and stored for 1 to 10 d before incubation was initiated. Egg weight remained less from collection through incubation for eggs from the mid-lay flock than those from the old flock. Fresh weight of early laid (first-in-sequence; C1) eggs was significantly greater than that for the middle-of-day laid (mid sequence; Cs), or late-in-day laid eggs (terminal-in-sequence; Ct). Percentage of egg weight loss during storage did not differ significantly between the mid-lay and old flocks but percentage of weight loss in the mid-lay flock was greater during incubation. Egg weight loss during storage of eggs from the middle-of-day laid (Cs) eggs was significantly greater than for early laid (C1) eggs, which was greater than for the late-in-day laid (Ct) eggs. Fertility was significantly decreased due to flock age but not due to oviposition time. Fertile hatchability was also significantly decreased due to flock age, but there was no significant effect of oviposition time. Early and late dead embryos increased with flock age, but there was no significant effect of oviposition time. It was concluded that there was no effect of oviposition time on fertility or fertile hatchability even though there were significant differences in egg weight and egg weight loss during storage due to oviposition time. PMID- 19903974 TI - Chronic renal failure: a neglected comorbidity of COPD. AB - BACKGROUND: To the best of our knowledge, the association between COPD and chronic renal failure (CRF) has never been assessed. Lean mass is frequently reduced in COPD, and the glomerular filtration rate (GFR) might be depressed in spite of normal serum creatinine (concealed CRF). We investigated the prevalence and correlates of both concealed and overt CRF in elderly patients with COPD. METHODS: We evaluated 356 consecutive elderly outpatients with COPD enrolled in the Extrapulmonary Consequences of COPD in the Elderly Study and 290 age-matched outpatients free from COPD. The GFR was estimated using the Modification of Diet in Renal Disease Study Group equation. Patients were categorized as having normal renal function (GFR > or = 60 mL/min/1.73 m(2)), concealed CRF (normal serum creatinine and reduced GFR), or overt CRF (increased serum creatinine and reduced GFR). Independent correlates of CRF were investigated by logistic regression analysis. RESULTS: The prevalence of concealed and overt CRF in patients with COPD was 20.8% and 22.2%, respectively. Corresponding figures in controls were 10.0% and 13.4%, respectively. COPD and age were significantly associated with both concealed CRF (COPD: odds ratio [OR] = 2.19, 95% CI = 1.17-4.12; age: OR = 1.06, 95% CI = 1.04-1.09) and overt CRF (COPD: OR = 1.94, 95% CI = 1.01-4.66; age: OR = 1.06, 95% CI = 1.04-1.10). Diabetes (OR = 1.96, 95% CI = 1.02-3.76), hypoalbuminemia (OR = 2.83, 95% CI = 1.70-4.73), and muscle-skeletal diseases (OR = 1.78, 95% CI = 1.01-3.16) were significant correlates of concealed CRF. BMI (OR = 1.05, 95% CI = 1.01-1.10) and diabetes (OR = 2.25, 95% CI = 1.26-4.03) were significantly associated with overt CRF. CONCLUSIONS: CRF is highly prevalent in patients with COPD, even with normal serum creatinine, and might contribute to explaining selected conditions such as anemia that are frequent complications of COPD. PMID- 19903975 TI - Social power and attitude strength over the life course. AB - Past findings indicate that middle-aged adults in the United States tend to be more resistant to attitude change than younger and older adults, but little is known about why this is so. The authors propose that midlife adults' disproportionate occupation of high-power social roles (which call for resoluteness) may partly explain their heightened resistance to persuasion. Using nationally representative data sets, the article first documents that in various domains the possession of social power peaks in midlife. It next documents that middle-aged adults place a high value on resoluteness, which suggests that they have internalized powerful role norms. Next, it shows that directly activating the concept of social power increases the perceived value of resoluteness. Finally, it demonstrates that the possession of powerful social roles partially mediates the relationship between age and resistance to persuasion. This work is the first to uncover a mechanism responsible for changes in attitude strength over the adult life course. PMID- 19903973 TI - Can we predict daily adherence to warfarin?: Results from the International Normalized Ratio Adherence and Genetics (IN-RANGE) Study. AB - BACKGROUND: Warfarin is the primary therapy to prevent stroke and venous thromboembolism. Significant periods of nonadherence frequently go unreported by patients and undetected by providers. Currently, no comprehensive screening tool exists to help providers assess the risk of nonadherence at the time of initiation of warfarin therapy. METHODS: This article reports on a prospective cohort study of adults initiating warfarin therapy at two anticoagulation clinics (university- and Veterans Affairs-affiliated). Nonadherence, defined by failure to record a correct daily pill bottle opening, was measured daily by electronic pill cap monitoring. A multivariable logistic regression model was used to develop a point system to predict daily nonadherence to warfarin. RESULTS: We followed 114 subjects for a median of 141 days. Median nonadherence of the participants was 14.4% (interquartile range [IQR], 5.8-33.8). A point system, based on nine demographic, clinical, and psychosocial factors, distinguished those demonstrating low vs high levels of nonadherence: four points or fewer, median nonadherence 5.8% (IQR, 2.3-14.1); five points, 9.1% (IQR, 5.9-28.6); six points, 14.5% (IQR, 7.1-24.1); seven points, 14.7% (IQR, 7.0-34.7); and eight points or more, 29.3% (IQR, 15.5-41.9). The model produces a c-statistic of 0.66 (95% CI, 0.61-0.71), suggesting modest discriminating ability to predict day level warfarin nonadherence. CONCLUSIONS: Poor adherence to warfarin is common. A screening tool based on nine demographic, clinical, and psychosocial factors, if further validated in other patient populations, may help to identify groups of patients at lower risk for nonadherence so that intensified efforts at increased monitoring and intervention can be focused on higher-risk patients. PMID- 19903976 TI - Implications of reflected appraisals of interpersonal insecurity for suspicion and power. AB - In three studies, the authors tested a model positing that chronically insecure individuals often believe that their relationship partners view them as highly insecure. In turn, because of expectations regarding the social consequences of expressing insecurities, these reflected appraisals of insecurity are thought to predict suspicion of partners' authenticity and feelings of powerlessness within relationships. Results supported these predictions. Self-esteem, attachment anxiety, neuroticism, proclivity for anger, and proclivity for hurt feelings predicted reflected appraisals of insecurity independently of whether partners detected insecurity. In turn, chronically insecure participants were suspicious of their partners' authenticity and felt powerless in their relationships partly because they believed they were viewed as insecure. This research suggests that beliefs that one is perceived as insecure, even when they are misguided, can partially explain interpersonal cognitions associated with actually being insecure. PMID- 19903978 TI - Slow walking speed in elderly people. PMID- 19903979 TI - Greater equality and better health. PMID- 19903980 TI - Slow walking speed and cardiovascular death in well functioning older adults: prospective cohort study. AB - OBJECTIVE: To study the relation between low walking speed and the risk of death in older people, both overall and with regard to the main causes of death. DESIGN: Prospective cohort study. SETTING: Dijon centre (France) of the Three City study. PARTICIPANTS: 3208 men and women aged >or=65 living in the community, recruited from 1999 to 2001, and followed for an average of 5.1 years. MAIN OUTCOME MEASURES: Mortality, overall and according to the main causes of death, by thirds of baseline walking speed (measured at maximum speed over six metres), adjusted for several potential confounders; Kaplan-Meier survival curves by thirds of baseline walking speed. Vital status during follow-up. Causes of death. Results During 16 414 person years of follow-up, 209 participants died (99 from cancer, 59 from cardiovascular disease, 51 from other causes). Participants in the lowest third of baseline walking speed had an increased risk of death (hazard ratio 1.44, 95% confidence interval 1.03 to 1.99) compared with the upper thirds. Analyses for specific causes of death showed that participants with low walking speed had about a threefold increased risk of cardiovascular death (2.92, 1.46 to 5.84) compared with participants who walked faster. There was no relation with cancer mortality (1.03, 0.65 to 1.70). In stratified analyses, cardiovascular mortality was increased across various strata defined by sex, median age, median body mass index (BMI), and level of physical activity. Conclusion Slow walking speed in older people is strongly associated with an increased risk of cardiovascular mortality. PMID- 19903982 TI - Differential effects of oral beta blockade on cardiovascular and sympathetic regulation. AB - In patients with hypertension, beta blockade decreases muscle sympathetic nerve activity (MSNA; micrographic technique) expressed in burst frequency (burst/min) but does not affect MSNA expressed in burst incidence (burst/100 heart beats), because reductions in blood pressure (BP) upon each diastole continue to deactivate the arterial baroreceptors, but at a slower heart rate (HR). We studied the effects of oral beta blockade on MSNA and baroreflex sensitivity (BRS) in normal participants. Bisoprolol (5 mg, 1 week) was administered in 10 healthy young adults, using a double-blind, placebo-controlled, randomized cross over study design. The beat-to-beat mean RR interval (RR) and systolic blood pressure (SBP) series were analyzed by power spectral analysis and power computation over the very low frequency (VLF), low frequency, and high frequency (HF) bands. Baroreflex sensitivity was computed from SBP and RR cross-analysis, using time and frequency domain methods. Bisoprolol increased RR (P < .0005), decreased mean SBP and diastolic blood pressure values (P < .01), did not change the SBP and RR powers, except for RR power in VLF (P < .02) and SBP power in HF (P < .03). The MSNA variability (P > .13) and respiratory pattern (P = .84) did not change from placebo to bisoprolol condition. The bisoprolol-induced bradycardia was associated with higher burst/100 heart beats (P < .05) and bisoprolol did not affect burst/min (P = .80). Time domain BRS estimates were increased after bisoprolol (P < .05), while frequency domain ones did not change (P > .1). Oral bisoprolol induces differential effects on sympathetic burst frequency and incidence in normal participants. Peripheral sympathetic outflow over time is preserved as a result of an increased burst incidence, in the presence of a slower HR. Unchanged BP and HR and MSNA variability suggests that the larger burst incidence is not due to sympathetic activation. PMID- 19903981 TI - Income inequality, mortality, and self rated health: meta-analysis of multilevel studies. AB - OBJECTIVE: To provide quantitative evaluations on the association between income inequality and health. DESIGN: Random effects meta-analyses, calculating the overall relative risk for subsequent mortality among prospective cohort studies and the overall odds ratio for poor self rated health among cross sectional studies. DATA SOURCES: PubMed, the ISI Web of Science, and the National Bureau for Economic Research database. Review methods Peer reviewed papers with multilevel data. Results The meta-analysis included 59 509 857 subjects in nine cohort studies and 1 280 211 subjects in 19 cross sectional studies. The overall cohort relative risk and cross sectional odds ratio (95% confidence intervals) per 0.05 unit increase in Gini coefficient, a measure of income inequality, was 1.08 (1.06 to 1.10) and 1.04 (1.02 to 1.06), respectively. Meta-regressions showed stronger associations between income inequality and the health outcomes among studies with higher Gini (>or=0.3), conducted with data after 1990, with longer duration of follow-up (>7 years), and incorporating time lags between income inequality and outcomes. By contrast, analyses accounting for unmeasured regional characteristics showed a weaker association between income inequality and health. Conclusions The results suggest a modest adverse effect of income inequality on health, although the population impact might be larger if the association is truly causal. The results also support the threshold effect hypothesis, which posits the existence of a threshold of income inequality beyond which adverse impacts on health begin to emerge. The findings need to be interpreted with caution given the heterogeneity between studies, as well as the attenuation of the risk estimates in analyses that attempted to control for the unmeasured characteristics of areas with high levels of income inequality. PMID- 19903983 TI - Neither K+ channels nor PI3K/Akt mediates the vasodilative effect of nebivolol on different types of rat arteries. AB - PURPOSE: Nebivolol is a highly selective beta(1)-adrenoceptor blocker with additional vasodilating properties. It has been shown that the nebivolol-induced vasorelaxation is nitric oxide (NO) dependent. The serine/ threonine protein kinase Akt phosphorylates endothelial cell NO synthase (eNOS) and enhances the ability of eNOS to generate NO. Previous studies have shown that the release of NO from the endothelium may be ascribed to the modulation of different types of K(+) channels. The current study was designed to determine whether K(+) channels or phosphatidylinositol-3-kinase (PI3K)/Akt may affect vasorelaxation induced by nebivolol in different rat arteries. METHODS: Rings of the rat aorta, carotid artery, femoral artery, and renal artery were suspended for isometric force recording. During contraction by KCl (60 mmol/L) or phenylephrine (PE; 10(-6) mol/L; femoral artery and renal artery were precontracted by 10(-5) mol/L), the effect of nebivolol (10(-7)-10(- 5) mol/L) was obtained in the presence of different potassium channel, PI3K/Akt, or NOS inhibitors. RESULTS: Nebivolol (10( 7)-10(-5) mol/L) relaxed precontractions induced by KCl and PE in different rat arteries, which was inhibited by the presence of the NOS inhibitor N(G)-nitro-L arginine methyl ester (L-NAME; 100 micromol/ L). The effect of nebivolol was concentration dependent. The exposure of the vessel rings to a selective inhibitor of PI3K wortmannin (5 x 10(-7) mol/L) or a selective inhibitor of Akt (1L-6-hydroxymethyl-chiro-inositol 2-(R)-2-O-methyl-3-O-octadecylcarbonate, 10( 5) mol/L) did not influence nebivolol-induced vasorelaxation. Similarly, K(+) channels blockers, iberiotoxin (100 nmol/L), glibenclamide (0.1 mmol/L), 4 aminopyridine (1 mmol/L), or BaCl(2) (1 mmol/L) had no influence on the relaxation of nebivolol in arteries precontracted by PE. CONCLUSION: Nebivolol produced a concentration-dependent vasodilation in different rat arteries precontracted by PE or KCl. In the isolated rat aorta, carotid artery, femoral artery, and renal artery, neither K(+) channels nor PI3K/Akt pathway was involved in the relaxation induced by nebivolol. PMID- 19903984 TI - Effect of targeting mitogen-activated protein kinase on cardiac remodeling in rats. AB - BACKGROUND: Increasing evidence suggests that the activation of p38 mitogen activated protein kinase (p38MAPK) plays a role in cardiac remodeling. Targeting p38MAPK using drugs reported to interfere with its phosphorylation, namely statins and all-trans retinoic acid (atRA), might play a role in ameliorating this remodeling. METHODS AND RESULTS: Cardiac remodeling was induced in male albino rats by chronic inhibition of nitric oxide (NO) synthesis by N-nitro L arginine methyl ester (L-NAME). Daily oral administration of L-NAME for 4 weeks resulted in the elevation of mean arterial blood pressure (MABP) together with cardiac remodeling evidenced by an increase in left ventricular-body weight ratio together with an increase in cardiac hydroxyproline concentration and a decrease in left ventricular papillary muscle-developed tension. An elevation in cardiac phosphorylated p38MAPK concentration, tumor necrosis factor alpha concentration and in cardiac caspase 3 activity was also observed. Administration of either rosuvastatin or all-trans retinoic acid (atRA), starting 4 weeks after L-NAME administration, ameliorated remodeling and improved all studied parameters. CONCLUSIONS: Targeting MAPK might represent a useful therapeutic avenue to ameliorate cardiac remodeling and support the notion that atRA and statins are potential candidates for the prevention and therapy of cardiac remodeling. PMID- 19903999 TI - Overview on 5-HT receptors and their role in physiology and pathology of the central nervous system. AB - The present review gives an overview on the serotonin (5-hydroxytryptamine; 5-HT) system, its receptors and their relationship to central nervous system physiology and disorders. Additionally, we also introduce the recent knowledge about the 5 HT receptor ligands in preclinical research, clinical trials and as approved drugs. PMID- 19904000 TI - Inhibitors of type 2 sodium glucose co-transporters--a new strategy for diabetes treatment. AB - In the last few years, the type 2 sodium glucose co-transporters (SGLT2) have been the subject of particular attention as a new, potent group of anti-diabetic drugs. SGLT2 inhibitors block glucose reabsorption in the kidneys, which prompts urinary excretion of glucose and results in lowering of its plasma levels. Although this group of medications is still under investigation, their efficacy in the treatment of type 2 diabetes mellitus (T2D) is very promising, with some of these inhibitors currently undergoing clinical trials. PMID- 19904001 TI - Characterization of the naturally occurring Arg344His variant of the human 5-HT 3A receptor. AB - The present study aimed at examining the function and pharmacological properties of the naturally occurring Arg344His variant of the human 5-HT(3A) receptor, identified in a schizophrenic patient. In intact human embryonic kidney (HEK) 293 cells expressing the wild-type (WT) or the variant receptor, the function was analyzed by indirect measurement of agonist-induced Ca(2+) current through the 5 HT(3A) receptor channel by an aequorin luminescence-based Ca(2+) assay. In cell membrane patches cation currents were determined electrophysiologically including technically demanding single channel analyses. The pharmacological properties were analyzed by [(3)H]GR65630 binding to cell membrane fragments. The density of [(3)H]GR65630 binding sites in cells expressing the variant receptor was reduced to 55% of that in cells expressing the WT receptor, which, however, was not accompanied by an analogous decrease in 5-HT-induced Ca(2+) influx through the receptor channel. However, the single channel analysis suggests an increase in single receptor channel mean open time (which is known to be subject of many variables) but not in unitary current amplitude. Radioligand competition experiments revealed that the affinity of five 5-HT(3) receptor agonists and four antagonists for the variant receptor did not differ from that for the WT receptor. In conclusion, the variant receptor resembles the WT receptor in that it forms functional homopentameric 5-HT(3A) receptors with identical pharmacological properties. In view of the lack of reduction in Ca(2+) flux through the variant receptor channels in spite of the decrease in its density on the cell membrane, the increase in single receptor channel mean open time appears to compensate for the reduction in variant receptor density. PMID- 19904002 TI - Treatment with 1,2,3,4,-tetrahydroisoquinoline affects glutamate release in the striatum but not the binding of [3H]MK-801 to NMDA receptors in the dopaminergic structures of the rat brain. AB - Overactivity of the glutamatergic system is thought to be closely related to the pathogenesis of Parkinson's disease. This study aimed to examine the effect of acute administration of 1,2,3,4-tetrahydroisoquinoline (TIQ), an endo- and exogenous amine suspected of inducing parkinsonism in humans on the release of glutamate in the striatum as well as to assess the impact of its chronic treatment on the binding of [(3)H]MK-801 to NMDA receptors in the dopaminergic structures of the rat brain. Striatal release of glutamate was measured in conscious, freely moving rats using a microdialysis method. [(3)H]MK-801 binding to NMDA receptors was examined in the striatum, nucleus accumbens and prefrontal cortex by autoradiographic method. TIQ administered acutely at a single dose of 100 mg/kg significantly decreased the level of extracellular glutamate, with the concentration decrease starting at 60 min and reaching minimum at 210 min after TIQ injection. TIQ administered chronically at the same dose for 3 weeks did not alter the binding of [(3)H]MK-801 to NMDA receptors in the examined dopaminergic structures at either 4 or 72 h after the last chronic injection. These results indicate that TIQ can modulate glutamate release in the striatum but that it does not affect the level of NMDA receptors in that structure. The latter data complete a list of recently published evidence that distinctly suggest that TIQ acts in the mammalian brain rather as a neuromodulator or even as a neuroprotective agent but not as a parkinsonism-inducing neurotoxin. PMID- 19904003 TI - Analgesic activity of 3-mono-substituted derivatives of dihydrofuran-2-one in experimental rodent models of pain. AB - Three derivatives of dihydrofuran-2-one (L-PP, L-PP1, and L-SAL) were administered by intraperitoneal injection and their analgesic activity was assayed in several models of pain. The activity of these derivatives were tested using a hot plate test, a writhing test, capsaicin- and glutamate-induced nociception, along with two models of local anesthesia, including a test for infiltration anesthesia in guinea pigs and the modified tail immersion test in mice. The results of these in vivo experiments show that these three derivatives of dihydrofuran-2-one possess analgesic activity in rodents. The ED(50) values of the tested compounds are lower or comparable to the ED(50) values of reference compounds (acetylsalicylic acid or morphine). For the most active derivative of dihydrofuran-2-one, L-PP1 (3-[4-(3-trifluoromethylphenyl)-piperazin-1-yl] dihydrofuran-2-one dihydrochloride), the ED(50) value was: 1.34 mg/kg, 0.79 mg/kg, 2.01 mg/kg and 3.99 mg/kg in the hot plate, writhing, capsaicin- and glutamate-induced pain tests, respectively. PMID- 19904004 TI - Effect of caffeine on the anticonvulsant effects of oxcarbazepine, lamotrigine and tiagabine in a mouse model of generalized tonic-clonic seizures. AB - Caffeine has been reported to be proconvulsant and to reduce the anticonvulsant efficacy of a variety of antiepileptic drugs (carbamazepine, phenobarbital, phenytoin, valproate and topiramate) in animal models of epilepsy and to increase seizure frequency in patients with epilepsy. Using the mouse maximal electroshock model, the present study was undertaken so as to ascertain whether caffeine affects the anticonvulsant efficacy of the new antiepileptic drugs lamotrigine, oxcarbazepine and tiagabine. The results indicate that neither acute nor chronic caffeine administration (up to 46.2 mg/kg) affected the ED(50) values of oxcarbazepine or lamotrigine against maximal electroshock. Similarly, caffeine did not modify the tiagabine electroconvulsive threshold. Furthermore, caffeine had no effect on oxcarbazepine, lamotrigine and tiagabine associated adverse effects such as impairment of motor coordination (measured by the chimney test) or long-term memory (measured by the passive avoidance task). Concurrent plasma concentration measurements revealed no significant effect on lamotrigine and oxcarbazepine concentrations. For tiagabine, however, chronic caffeine (4 mg/kg) administration was associated with an increase in tiagabine concentrations. In conclusion, caffeine did not impair the anticonvulsant effects of lamotrigine, oxcarbazepine, or tiagabine as assessed by electroconvulsions in mice. Also, caffeine was without effect upon the adverse potential of the studied antiepileptic drugs. Thus caffeine may not necessarily adversely affect the efficacy of all antiepileptic drugs and this is an important observation. PMID- 19904005 TI - Anti-apoptotic effect of memantine against staurosporine- and low-potassium induced cell death in cerebellar granule cells: a development-dependent effect. AB - Memantine, a NMDA receptor antagonist used in several experimental models of neuronal cell injury, is a neuroprotective agent that can attenuate neuronal apoptosis connected with over-stimulation of NMDA receptors. In the present study, we evaluated the impact of memantine on apoptosis in primary cerebellar granule cell (CGC) cultures at 7 and 12 day in vitro (DIV). Cell death was induced by staurosporine (St, 0.5 microM) or by decreasing the level of potassium in the culture medium (LP, 5 mM KCl). Both treatments induced cell death in CGC with higher cell-damaging effects at 12 DIV and 7 DIV neurons for St and LP, respectively. Memantine (0.1-2 microM) partially attenuated St-induced apoptosis only in 7 DIV CGC as assessed by DNA fragmentation and LDH release, but not caspase-3 activity. During LP-induced apoptosis, memantine decreased LDH release and DNA fragmentation, but not affected caspase-3 activity in 7 and 12 DIV CGC. Interestingly, we found no beneficial effects of other NMDA antagonists, including a competitive antagonist such as AP-5 (100 microM) and an uncompetitive antagonist such as MK-801, (1 microM). In conclusion, our data suggest that the anti-apoptotic effects of memantine in CGC are developmentally regulated and its neuroprotective action occurs through an NMDAR-independent mechanism. PMID- 19904006 TI - Commonly used ryanodine receptor activator, 4-chloro-m-cresol (4CmC), is also an inhibitor of SERCA Ca2+ pumps. AB - 4-Chloro-m-cresol (4CmC) is an extensively used activator of ryanodine receptors (RyRs). Studies have shown that 4CmC, at a concentration of 1 mM, is sufficient to cause Ca(2+) release through RyRs. Here, we show that mM concentrations of 4CmC also inhibit the sarcoplasmic-endoplasmic reticulum Ca(2+)-ATPase (SERCA), (IC(50) 2-3mM) and cause Ca(2+) release. 4CmC also causes increased intracellular [Ca(2+)] levels in COS-7 cells, which lack functional RyRs. Thus, any increase in [Ca(2+)] levels associated with use of 4CmC (>or= 1mM) could lead to non-specific Ca(2+) changes due to SERCA inhibition rather than RyR activation. PMID- 19904007 TI - Influence of ABCB1 gene polymorphisms on the pharmacokinetics of azithromycin among healthy Chinese Han ethnic subjects. AB - The aim of this study was to evaluate the effects of ABCB1 gene polymorphisms on azithromycin pharmacokinetics in Chinese Han ethnic subjects. In total, 20 healthy volunteers with various ABCB1 genotypes (6 with 2677GG/3435CC, 8 with 2677GT/3435CT, 6 with 2677TT/3435TT) were enrolled. Each was given a single oral dose of 500 mg azithromycin. Plasma concentration was measured for up to 96 h by LC/MS/MS. As shown, C(max) was significantly lower among individuals with 2677TT/3435TT genotype (468.0 +/- 173.4 ng x h/ml) than those with 2677GG/3435CC (911.2 +/- 396.4 ng x h/ml, p = 0.013). However, the t(max) value was higher among subjects with 2677TT/3435TT (2.0 +/- 0.5 h) than those with 2677GT/3435CT (1.6 +/- 0.3 h) or 2677GG/3435CC (1.4 +/- 0.4 h) genotypes (p = 0.068 and p = 0.026, respectively). Furthermore, the AUC(last) tended to be higher among subjects with 2677GG/3435CC than those with 2677GT/3435CT or 2677TT/3435TT genotypes (5000.2 +/- 1610.0 vs. 4558.0 +/- 805.0 vs. 4131.0 +/- 995.1 ng/ml). Our results showed for the first time that azithromycin pharmacokinetics may be influenced by particular polymorphisms of the ABCB1 gene. Individualized dosage regimen design incorporating such information may improve the efficacy of the drug while reducing adverse reactions. PMID- 19904008 TI - Perazine at therapeutic drug concentrations inhibits human cytochrome P450 isoenzyme 1A2 (CYP1A2) and caffeine metabolism--an in vitro study. AB - The aim of the present study was to estimate the inhibitory effect of perazine, a phenothiazine neuroleptic with piperazine structure in a side chain, on human CYP1A2 activity measured as a rate of caffeine 3-N- and 1-N-demethylation. Moreover, the influence of perazine on other caffeine metabolic pathways such as 7-N-demethylation (CYP1A2, CYP2C8/9, CYP3A4) and 8-hydroxylation (CYP3A4, CYP1A2, CYP2C8/9) was also determined. The Dixon analysis showed that in both human liver microsomes and Supersomes CYP1A2 perazine potently and to a similar degree inhibited caffeine 3-N-demethylation (K(i) = 3.5 microM) and 1-N-demethylation (K(i) = 5 microM). Perazine moderately diminished the rate of caffeine 7-N demethylation in Supersomes CYP1A2 (K(i) = 11.5 microM) and liver microsomes (K(i) = 20 microM), and attenuated C-8-hydroxylation (K(i) = 15.5 microM) in Supersomes CYP1A2. On the other hand, perazine weakly inhibited caffeine C-8 hydroxylation in liver microsomes (K(i) = 98 microM). About 80% of basal CYP1A2 activity was reduced by the therapeutic concentrations of perazine (5-10 microM). The obtained results show that perazine at its therapeutic concentrations is a potent inhibitor of human CYP1A2. Hence, taking account of CYP1A2 contribution to the metabolism of endogenous substances (steroids), drugs (xanthine derivatives, phenacetin, propranolol, imipramine, phenothiazine neuroleptics, clozapine) and carcinogenic compounds, the inhibition of CYP1A2 by perazine may be of physiological, pharmacological and toxicological importance. PMID- 19904009 TI - Effects of ischemia-reperfusion and pretreatment with mildronate on rat liver mitochondrial function. AB - Mildronate (3-(2,2,2-trimethylhydrazinium) propionate), which is mostly used in cardiological practice and is considered an anti-ischemic drug, was designed to inhibit carnitine biosynthesis in order to prevent accumulation of cytotoxic intermediate products of fatty acid beta-oxidation. Recently it was shown that the mitochondrial respiratory chain may also be a target for mildronate action. In this study, we aimed to investigate whether mildronate can protect the liver against a 90-min normothermic ischemia/30-min reperfusion-induced mitochondrial dysfunction. Rats were pre-treated for one or two weeks with mildronate (100 mg/kg/day or 200 mg/kg/day) or Ringer solution and subjected to ischemia/reperfusion.We found that ischemia/reperfusion caused a decrease in mitochondrial State 3 respiration rate and in the respiratory control index (RCI), and an increase in State 2 respiration rate with succinate, glutamate + malate and palmitoyl-L-carnitine + malate. One or two weeks of pre-treatment of rats with different doses of mildronate did not reduce the ischemia/reperfusion induced decrease in the State 3 respiration rate or RCI; however, a one week pre treatment slightly diminished the increase in the State 2 respiration rate with glutamate + malate substrates. The leakage of the liver enzymes, aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase, was similar in both the untreated and pre-treated with mildronate groups. No steatotic livers were observed in any experimental groups after mildronate pre-treatment. In conclusion, 90 min of liver ischemia followed by a 30 min reperfusion has a deleterious effect on rat liver mitochondrial function. Mildronate pre-treatment of rats at doses of 100 or 200 mg/kg/day for one or two weeks did not prevent ischemia/reperfusion-induced mitochondrial dysfunction and liver injury. PMID- 19904010 TI - Chronic beta-adrenergic activation-induced left ventricular systolic dysfunction is associated with systemic release of TNF-alpha and IL-1-beta in rats. AB - The relationship between systemic cytokine release and chronic beta-adrenergic activation-induced left ventricular dysfunction (LVSD) has not been widely reported in the literature. In the present study,we examined changes in the serum levels of inflammatory cytokines (IL-1-beta, IL-6 and TNF-alpha) following chronic beta-adrenergic activation-induced LVSD. Male Wistar rats were administered isoproterenol (ISO, 5 mg/kg, sc, once daily) for 4 weeks (ISO 4) or 12 weeks (ISO 12). Echocardiography was done and serum levels of IL-1-beta, IL-6 and TNF-alpha were estimated at the end of each protocol. In the ISO 4 group, there was a significant increase in relative wall thickness (p < 0.01) and heart weight: body weight ratio (p < 0.001) without any significant changes in fractional shortening (FS) or serum cytokine levels. However, in the ISO 12 group, there was a 32% decrease in FS along with increased serum levels of IL-1 beta and TNF-alpha. The present findings indicate that LVSD induced by chronic beta-adrenergic activation in rats is accompanied by increased serum cytokine levels. PMID- 19904012 TI - Possible new organoselenium supplement--evaluation of its influence on the kidneys in comparison with inorganic sodium selenite. AB - The aim of this work was to evaluate the possibility of using of the new selenoorganic ring compound, 3-(o-chlorobenzoylamino)-2-(o-tolylimino)-4-methyl-4 selenazoline, as a selenium supplement by investigating the influence of its short-term administration on Se accumulation and antioxidant status in kidney. For 10 days, adolescent male Wistar rats were treated with saline (control group), Na(2)SeO(3) (Se-IN group) or the studied compound (Se-ORG group) (5 x 10( 4) mg Se/g of once a day) via a stomach tube. The selenium concentration, total antioxidant status (TAS), activities of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx), concentrations of ascorbic acid (AA) and reduced glutathione (GSH) and concentration of malonyldialdehyde (MDA) were determined in the kidney homogenates. TAS was significantly reduced in the Se-ORG group compared to the control. Reduced glutathione was markedly decreased in Se-treated animals compared to the control and in the Se-ORG group compared to the Se-IN group. Malonyldialdehyde was significantly increased in the Se supplemented groups compared to the control group but considerably less so in the Se-ORG group. All other studied parameters displayed no significant differences. No increase in the accumulation of selenium and the partial impairment of the antioxidant status and enhancement of lipid peroxidation in the kidneys resulting from Se treatment could suggest that in the first period of administration, excess selenium was excreted with urine, leading to a disturbance of kidney functions. Comparison of the effect of our compound with that exerted by inorganic Na(2)SeO(3) suggests that the studied compound could be considered as a possible supplement after further investigations, including determination of selenium excretion with urine, as well as repetition of this study using a wide range of doses and periods of supplementation. PMID- 19904011 TI - Enhancement of fibrinolysis by gel-filtered platelets and its quenching by cytochalasin B and GPIIb/IIIa antagonists. AB - The effects of gel-filtered platelets on euglobulin clot lysis time (ECLT) were analyzed to elucidate the possible role of platelets in thrombolysis. Gel filtered platelet-supplemented ECLT (plt-ECLT) was significantly shorter than ECLT without platelets (regular ECLT). Abciximab, anti-glycoprotein IIb/IIIa (GPIIb/IIIa) antibody, and cytochalasin B nullified the enhancement of ECLT by platelets, and increased plt-ECLT beyond regular ECLT. When gel-filtered platelets were used after disruption, ECLT was not shortened but rather became longer than regular ECLT, probably due to natural fibrinolysis inhibitors released from platelets. Therefore, for platelets to enhance fibrinolysis, intact cell structure and cytoskeletal reorganization after thrombin stimulation is required. Various GPIIb/IIIa antagonists prolonged plt-ECLT. The concentrations of GPIIb/IIIa antagonists required to prolong plt-ECLT, were varied. Interestingly, the effects of these antagonists were independent of their ability to inhibit thrombin-induced platelet aggregation, but dependent on their ability to induce clot retraction. T-250, a GPIIb/IIIa antagonist, had the smallest effect on plt-ECLT. These drugs do not affect regular ECLT or tissue plasminogen activator (tPA)-catalyzed Glu-plasminogen activation in the presence of thrombin activated platelets. Although their overall effect on thrombolysis is inhibitory, platelets could promote fibrinolysis through a GPIIb/IIIa-dependent mechanism. PMID- 19904013 TI - Effect of atorvastatin on paraoxonase1 (PON1) and oxidative status. AB - It has been proposed that paraoxonase1 (PON1), a high density lipoprotein (HDL) associated esterase/lactonase, has anti-atherosclerotic properties. The activity of PON1 is influenced by PON1 polymorphisms. However, the influence of PON1 polymorphisms on PON1 activity and oxidative stress in response to lipid-lowering drugs remains poorly understood. The objective of the present study was to investigate the effects of atorvastatin on PON1 activity and oxidative status. The influence of PON1 polymorphisms on PON1 activity and oxidative status in response to atorvastatin treatment was also evaluated. In total, 22 hypercholesterolemic patients were treated with atorvastatin at a dose of 10 mg/day for 3 months. Lipid profile, lipid oxidation markers (malondialdehyde (MDA), conjugated diene (CD), total peroxides (TP)), total antioxidant substance (TAS), oxidative stress index (OSI), and paraoxonase1 activity were determined before and after treatment. L55M, Q192R, and T(-107)C PON1 polymorphisms were also determined. Atorvastatin treatment significantly reduced the levels of total cholesterol (24.5%), low density lipoprotein (LDL) cholesterol (25.4%), triglycerides (24.4%), CD (4.4%), MDA (15.2%), TP (13.0%) and OSI (24.0%), and significantly increased the levels of TAS (27.3%), and PON1 activity (14.0%). Interestingly, the increase in PON1 activity and the reduction in oxidative stress in response to atorvastatin were influenced only by the PON1 T-107C polymorphism. Atorvastatin treatment improved the lipid profile, lipid oxidation, and oxidative/antioxidative status markers including the activity of PON1 towards paraoxon. These beneficial effects may be attributed to the antioxidant properties of statins and the increase in PON1 activity. The increase in PON1 activity was enhanced by the PON1 T-107C polymorphism. PMID- 19904014 TI - Antihyperglycemic effects of ASP8497 in streptozotocin-nicotinamide induced diabetic rats: comparison with other dipeptidyl peptidase-IV inhibitors. AB - (2S,4S)-4-Fluoro-1-({[4-methyl-1-(methylsulfonyl)piperidin-4 yl]amino}acetyl)pyrrolidine-2-carbonitrile monofumarate (ASP8497) is a novel dipeptidyl peptidase (DPP)-IV inhibitor. In this study, we investigated the antidiabetic potency, mechanism, and duration of action of ASP8497 both in vitro and in vivo, and compared it with the DPP-IV inhibitors vildagliptin, sitagliptin, and saxagliptin. ASP8497 inhibited rat plasma DPP-IV activity in vitro with an IC(50) value of 2.96 nmol/l, while those for vildagliptin, sitagliptin, and saxagliptin were 2.12, 8.98, and 2.00 nmol/l, respectively. In rats that had streptozotocin-nicotinamide-induced, mildly diabetes, oral administration of ASP8497 dose-dependently and sustainably inhibited plasma DPP IV activity. In addition, ASP8497 dose-dependently and significantly improved glucose tolerance with a concomitant increase in plasma glucagon-like peptide 1 (GLP-1) and insulin levels at both 0.5 h and 8 h after dosing. The order of both potency and duration of action for plasma DPP-IV inhibition and glucose tolerance improvement was as follows: saxagliptin > ASP8497 = vildagliptin = sitagliptin. These results suggest that ASP8497 exerts a potent and long-acting DPP-IV inhibitory effect and improves glucose tolerance through glucose-dependent insulinotropic action via elevation of the GLP-1 level in streptozotocin nicotinamide-induced mildly diabetic rats. This compound is expected to be useful as a therapeutic agent for impaired glucose tolerance and type 2 diabetes. PMID- 19904015 TI - Effects of high-fat mixed-lipid diet and exercise on the antioxidant system in skeletal and cardiac muscles of rats with colon carcinoma. AB - Epidemiological and experimental studies suggest that eating habits and a sedentary lifestyle play a critical role in the incidence of colon carcinoma. In order to investigate the effects of high-fat mixed-lipid (HFML) diet in conjunction with long-term swimming, the antioxidant capacity of skeletal and cardiac muscles were observed in rats with 1,2-dimethylhydrazine (DMH)-induced colon carcinoma. Male Wistar rats were randomly divided into one control group and four cancer groups: sedentary and swimming groups fed low fat corn oil diet and sedentary and swimming groups, fed a HFML diet. After 6 months of swimming, rats were sacrificed and the blood, cardiac and soleus muscle were taken for analysis. Serum cholesterol, triglyceride and glucose concentrations were measured and the activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase as well as levels of malondialdehyde and glutathione were determined. The results showed that endurance swimming prevented lipid peroxidation in the soleus muscle of HFML diet rats due to elevated activities of antioxidant enzymes. On the other hand, increased lipid peroxidation in the hearts of all cancer groups indicated that DMH-induced colon carcinoma impaired the antioxidant status of the heart. This failure in heart tissue indicated that enhanced antioxidant capacity after regular physical activity is not sufficient to offset oxidative stress caused by DMH-induced colon carcinoma. PMID- 19904016 TI - Erythropoietin inhibits liver gelatinases during galactosamine-induced hepatic damage in rats. AB - Matrix metalloproteinase (MMP)-2 and -9 (gelatinases) participate in extracellular protein remodeling. Moreover, they are involved in the development of hepatic fibrosis. The goal of this study was to evaluate liver gelatinase activities after erythropoietin (Epo) treatment (1U/dose, sc) in experimentally damaged livers of rats treated with D-galactosamine (Gal, 800 mg/kg/dose, ip). Sixty rats were divided into six equal groups: I - received 5 doses of Epo and a single dose of Gal [the experiment duration (ED): 10 days]; II - received 5 doses of Epo and 3 doses of Gal (ED: 14 days); III - received only 5 doses of Epo (ED: 9 days); IV - received 3 doses of Gal (ED: 5 days);V - received a single dose of Gal (ED: 1 day); VI - control group (ED: 9 days). The animals were sacrificed and the livers were collected 48 h after the last drug administration. The activity of gelatinases was measured using gelatin zymography. No fluctuations in gelatinase activities were observed after the administration of a single dose of Gal in comparison to the control group. However, a significant increase in gelatinase activities was observed after treatment with three doses of Gal. Five doses of Epo administrated before Gal treatment prevented elevated gelatinase activities: MMP-9 activity was comparable to control, and MMP-2 activity was decreased (group II). The gelatinase activities was lower in group I and II in comparison to the control group. These results revealed that Epo decreases MMP-2 and MMP-9 activity, suggesting that it is a hepatoprotective agent against hepatic damage induced by galactosamine injection. PMID- 19904017 TI - Effect of repeated co-treatment with fluoxetine and amantadine on the behavioral reactivity of the central dopamine and serotonin system in rats. AB - In the present study we found that repeated co-treatment with fluoxetine and amantadine for 14 days (but not for 7 days) enhanced the hyperactivity induced by amphetamine or quinpirole (a dopamine D(2/3) agonist), compared to treatment with either drug alone. Whereas repeated co-treatment with fluoxetine and amantadine for 7 days more potently inhibited the behavioral syndrome evoked by the 5 hydroxytryptamine (5-HT)(1A) receptor agonist (+/-)-8-hydroxy-2(di-n-propylamino) tetralin hydrobromide (8-OH-DPAT), it did not change the action of the 5-HT(2) receptor agonist (+/-)-1-(4-iodo-2,5-dimethoxyphenyl)-2-aminopropane hydrochloride (/+/-/-DOI). The obtained results support the hypothesis that repeated co-treatment with fluoxetine and amantadine may evoke more effective antidepressant activity than treatment with fluoxetine alone. Moreover, our results suggest that 5-HT(1A) receptors are useful targets for the development of more rapidly acting and more effective medication. PMID- 19904018 TI - Lack of effect of sildenafil on cocaine-induced convulsions in mice. AB - The convulsant action of cocaine and the proconvulsant effects of sildenafil, a drug which is widely used in the treatment of erectile dysfunction, have been documented both in humans and mice. Since it was reported that sildenafil alone, and in conjunction with cocaine, is used recreationally, the present study was performed to examine the influence of sildenafil on cocaine-induced seizures in mice.We showed that administration of sildenafil (5-20 mg/kg, ip) did not affect latency to clonic seizures induced by ip administration of cocaine at a dose of 85 mg/kg, nor did it influence seizure incidence and mortality.We conclude that sildenafil does not significantly increase the risk of seizures when co administered with cocaine. PMID- 19904019 TI - Effect of pregnancy and tobacco smoke on the antioxidant activity of rutin in an animal model. AB - Tobacco smoke is a source of free radicals and causes oxidative stress in smokers' tissues. The aim of the current study was to evaluate the effect of rutin on the total antioxidant status (TAS) in pregnant and non-pregnant rats that were exposed to cigarette smoke. TAS in brain, lungs, liver, kidneys and plasma were measured by the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) radical-cation decolorization assay. In pregnant rats, a diversified distribution of endogenous antioxidants was found in comparison to the matched non-pregnant animals. In pregnant rats, TAS was higher in plasma (by 33%) and kidney (by 76%), and lower in brain (by 48%) and liver (by 50%) compared with non pregnant rats. Generally (except liver), exposure to tobacco smoke caused an increase in the antioxidative status of pregnant compared to non-pregnant animals (by 29, 16, 18 and 87% in plasma, brain, lung and kidney, respectively). Overall, rutin had little (plasma, non-pregnant rats) or a no protective effect in the examined tissues. PMID- 19904020 TI - Effects of N-acetylcysteine and ebselen on arachidonic acid release from astrocytes and neurons cultured in normoxic or simulated ischemic conditions. AB - Arachidonic acid (AA) is released from cells after nervous tissue injuries.We treated rat cortical neurons and astrocytes cultured under normoxic or simulated ischemic conditions with N-acetylcysteine (100 or 200 microM) or ebselen (10 or 20 microM). N-acetylcysteine decreased AA release in normoxic astrocytes, while ebselen decreased AA release from astrocytes in both conditions. N-acetylcysteine produced no changes in neuronal AA release. A low dose of ebselen significantly increased AA release from neurons in both conditions. The influence of N acetylcysteine and ebselen on AA release might be implicated in their effects on astrocytes and neurons, however, the exact mechanisms have yet to be explained. PMID- 19904021 TI - Frequency of common CYP3A5 gene variants in healthy Polish newborn infants. AB - Cytochrome P450 monooxygenases catalyze the metabolism of approximately 40-60% of widely used drugs with a A6986G CYP3A5 polymorphism determining expresser (A6986, *1) and reduced- expresser (*3) variants with modified drug metabolism activity. In this report, the allele frequency of CYP3A5 *1 and *3 (A6986 or G6986, respectively) was analyzed by the PCR-RFLP technique in a cohort of 200 Polish newborns from the West Pomeranian region. Of the studied group, 1% (n = 2/200) proved homozygous for the CYP3A5*1 allele, 89% (n = 178/200) for the *3 allele, and 10% (n = 20/200) were heterozygous for *1/*3. Similar frequencies were found in other Caucasian European populations. This study provides basic genetic data related to the metabolism of drugs, with a narrow therapeutic window in a Polish population. PMID- 19904022 TI - Stability and analysis of configuration-tunable bi-directional MWNT bearings. AB - We report on the energetic and structural stability of configuration-tunable, bi directional linear bearings based on cap-less, partial segments engineered within individual multi-walled carbon nanotubes (MWNTs). Using computational models, we show that an externally applied excitation force can be used to select an operating bearing configuration with a desired stiffness and operating frequency. Our models also demonstrate the possibility of simultaneous, independent operation of multiple bearings within a single NT segment, paving the way towards ultra-high device densities with molecular-scale footprints. PMID- 19904023 TI - Synthesis and characterization of silver-nanoparticle-impregnated fiberglass and utility in water disinfection. AB - A number of researchers have deployed silver (Ag) nanoparticles through a number of techniques on various substrates including carbon, zeolites and polymers for water disinfection applications. However, Ag impregnated on an inorganic fiberglass surface through a simple electroless process was only recently reported for the first time. Fiberglass impregnated with Ag nanoparticles displays superior performance over carbon-based silver support systems but little is known about the factors that affect the architecture of the system, its interfacial properties and its consequent bactericidal activity. In this study, Ag content and particle size on a fiberglass substrate were manipulated by adjusting the AgNO(3) concentration, immersion time, temperature, solution pH and reduction temperature. The reduction chemistry of the Ag-nanoparticle-impregnated fiberglass is described and supported with thermal gravimetric analysis (TGA) and photoelectron spectroscopy (XPS) measurements. The Ag content along with the particle size and particle size distribution were characterized with scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy dispersive x-ray spectroscopy (EDS) and x-ray diffraction (XRD). The Ag content on the fiberglass mats ranged from 0.04 to 4.7 wt% Ag/g-fiber with a size distribution of 10-900 nm under standard processing conditions. Inductively coupled plasma mass spectrometry (ICP-MS) was used to analyze the Ag desorption from the fiberglass substrate, while the bactericidal properties were evaluated against Escherichia coli (E. coli). PMID- 19904024 TI - Scanning quantum decoherence microscopy. AB - The use of qubits as sensitive nanoscale magnetometers has been studied theoretically and recently demonstrated experimentally. In this paper we propose a new concept, in which a scanning two-state quantum system is used to probe a sample through the subtle effects of decoherence. Mapping both the Hamiltonian and decoherence properties of a qubit simultaneously provides a unique image of the magnetic (or electric) field properties at the nanoscale. The resulting images are sensitive to the temporal as well as spatial variation in the fields created by the sample. As examples we theoretically study two applications; one from condensed matter physics, the other biophysics. The individual components required to realize the simplest version of this device (characterization and measurement of qubits, nanoscale positioning) have already been demonstrated experimentally. PMID- 19904025 TI - Aligned AlN nanowires by self-organized vapor-solid growth. AB - Highly oriented AlN single crystal nanowires with aspect ratio up to 600, diameter in the range of 40-500 nm, and 100 microm lengths, have been synthesized via a vapor-solid growth mechanism. The results were obtained at 1750 degrees C and 850 mbar nitrogen pressure on vicinal SiC substrates pretreated by SiC sublimation epitaxy in order to attain distinguishable terraces. It was found that the nanowires change in thickness after they have reached a critical length, and this fact contributes to an understanding of the growth mechanism of AlN nanowires. The nanowires are hexagonally shaped and perfectly aligned along the [0001] direction with a small tilt given by the substrate vicinality. Under nitrogen excess a preferential growth along the c-axis of the wurtzite structure takes place while below some critical value of nitrogen pressure the growth mode switches to lateral. The AlN nanowires are shown to have a dislocation free wurtzite crystal structure. Some possible applications are discussed. PMID- 19904026 TI - InSb heterostructure nanowires: MOVPE growth under extreme lattice mismatch. AB - We demonstrate the growth of InSb-based nanowire heterostructures by metalorganic vapour phase epitaxy and use it to integrate InSb on extremely lattice-mismatched III-V nanowire templates made of InAs, InP, and GaAs. Influence of temperature, V/III ratio, and diameter are investigated in order to investigate the growth rate and morphology. The range of growth temperatures used for InSb nanowire growth is very similar to that used for planar growth due to the nature of the precursor decomposition. This makes optimization of growth parameters very important, and more difficult than for most other nanowire III-V materials. Analysis of the InSb nanowire epitaxial quality when grown on InAs, InP, and GaAs, along with InSb segment and particle compositions are reported. This successful direct integration of InSb nanowires, on nanowire templates with unprecedented strain levels show great promise for fabrication of vertical InSb devices. PMID- 19904027 TI - Shaping nano-/micro-particles for enhanced vascular interaction in laminar flows. AB - Non-spherical nano-/micro-particles can drift laterally (hydrodynamic margination) in a linear laminar flow under the concurrent effect of hydrodynamic and inertial forces. Such a feature can be exploited in the rational design of particle-based intravascular and pulmonary delivery systems and for designing new flow fractioning systems for high-throughput particle separation. A general approach is presented to predict the marginating behavior of non-spherical particles. The lateral drift velocity is shown to depend on the particle Stokes number St(a) and to grow with the size, density and rotational inertia of the particle. Elongated particles, in particular, low aspect ratio discoidal particles, exhibit the largest propensity to marginate in a linear laminar flow. In the blood microcirculation, at low shear rates (S<100 s(-1)), non-spherical particles oscillate around their trajectory and margination can only be achieved through the application of external force fields (gravitational, magnetic); whereas for larger S (100 s(-1)or=5 mm) in 21 patients. In detecting and grading GVs, there were moderate agreements (kappa value: 0.514 to 0.563) between MDCT and EGD, but in differentiating large varices requiring prophylactic therapy, a substantial agreement (kappa value: 0.804 for radiologist 1 and 0.796 for radiologist 2) was found. For radiologist 1, the sensitivity, specificity, accuracy, and positive and negative predictive values of MDCT for the identification of large GVs were 85.7%, 96.2%, 94.5%, 81.8%, and 97.1%, respectively; whereas for radiologist 2, they were 81.0%, 97.2%, 94.5%, 85.0%, and 96.3%, respectively. In evaluating the afferent and efferent veins of varices, the sensitivity, specificity, accuracy, and positive predictive value of MDCT portography were more than 80.0%. CONCLUSIONS: MDCT is an effective screening tool for differentiating large GVs and revealing the afferent and efferent veins of varices in patients with cirrhosis. PMID- 19904220 TI - Decomposing N2 NOGO wave of event-related potentials into independent components. AB - Inconsistencies in previous attempts to localize the N2 wave in the GO/NOGO task led to the present investigation. The inconsistencies were probably because of heterogeneity of psychological operations involved in GO/NOGO tasks. We applied the independent component analysis to a collection of individual event-related potentials in response to GO and NOGO cues in the two stimulus visual GO/NOGO task. The selected six independent components with different topographies and time courses constituted 87% of the artifact-free signal variance. Three of them were loaded into the frontally distributed N2 wave. According to standardized low resolution electromagnetic tomography these three independent components were generated in the supplementary motor cortex, left angular gyrus and anterior cingulate cortex. PMID- 19904221 TI - Lipomatous pseudohypertrophy of the pancreas: a clinicopathologically distinct entity. AB - OBJECTIVES: Owing to the challenges in obtaining pancreatic biopsies, pancreatic resection for presumed malignancy is often performed without histological confirmation. As a result, benign lesions are sometimes surgically removed. One such condition, which is poorly defined in the literature, is referred to as lipomatous pseudohypertrophy (LPH) of the pancreas. METHODS: Five cases of LPH were analyzed. RESULTS: Four patients underwent surgical resection, 3 of which were diagnosed preoperatively by radiology as having ductal adenocarcinoma. The fourth case was correctly interpreted by magnetic resonance imaging as LPH, but the patient underwent resection because of the intractable pain due to pancreatitis. The fifth patient has been placed on watchful waiting.Two tumors were in the pancreatic head, one in the tail, one in the uncinate process, and one demonstrated diffuse involvement. Microscopically, they were characterized as having normal lipocytes without lipoblasts or inflammation. Within the adipose tissue, scattered microscopic foci of pancreatic parenchyma could be seen. CONCLUSION: Lipomatous pseudohypertrophy of the pancreas is a distinct entity characterized by localized/diffuse replacement of pancreatic parenchyma with mature adipose tissue. It forms a pseudotumor that may be difficult to distinguish clinically from pancreatic adenocarcinoma. This entity should be considered when evaluating patients with a new diagnosis of a hypodense pancreatic neoplasm on imaging. PMID- 19904222 TI - Protection against chronic pancreatitis and pancreatic fibrosis in mice overexpressing pancreatic secretory trypsin inhibitor. AB - OBJECTIVES: Mutations in the gene encoding for pancreatic secretory trypsin inhibitor (PSTI) can contribute to chronic pancreatitis. In the current study, we tested whether overexpression of PSTI-I in mice protects against chronic pancreatitis and pancreatic fibrosis. METHODS: Rat PSTI-I expression was targeted to pancreatic acinar cells in transgenic mice. Chronic pancreatitis was achieved by intraperitoneal injection of cerulein for 10 weeks. Pancreatitis severity was assessed by histological grading of inflammatory infiltrate, atrophy, and fibrosis; quantitation of myeloperoxidase (MPO) activity; quantitative morphometric analysis of collagen content; and measurements of type I collagen, fibronectin, and transforming growth factor beta mRNA expression. RESULTS: Cerulein administration to nontransgenic mice produced histological evidence of inflammatory infiltrate, glandular atrophy, and parenchymal fibrosis and increased collagen production, MPO activity, and collagen I and fibronectin mRNA levels. In cerulein-treated PSTI transgenic mice, there were significant reductions in inflammatory infiltrate, MPO activity, fibrosis, and collagen I and fibronectin mRNA levels. Transgenic mice treated with cerulein had significantly less collagen than nontransgenic mice. CONCLUSIONS: The severity of chronic pancreatitis and pancreatic fibrosis is significantly reduced in mice expressing rat PSTI-I. We propose that pancreatic trypsin inhibitors play a protective role in the pancreatic response to repeated injurious events. PMID- 19904223 TI - Profiling pancreatic cancer-secreted proteome using 15N amino acids and serum free media. AB - OBJECTIVES: A new method of determining protein turnover by labeling protein with N amino acids was used in conjunction with serum-free cell culture to profile secreted proteins that are released by MIA PaCa-2 pancreatic cancer cells in culture. METHODS: MIA PaCa-2 cells were first cultured in Dulbecco modified Eagle medium (Gibco by Invitrogen, Carlsbad, Calif) with 10% fetal bovine serum, then in serum-free modified Eagle medium with or without 50% N algal amino acid mixture. The effect of oxythiamine chloride on secreteome was studied. Secreteome from cell culture media was analyzed by 2-dimensional (2D) gel electrophoresis. Differentially expressed proteins were detected and identified. Protein turnover rates were calculated according to the newly established method. Western blot and enzyme-linked immunosorbent assay were used to validate identified proteins. RESULTS: Among the 14 differentially expressed proteins after oxythiamine treatment, tissue inhibitor of metalloproteases-1 and cytokeratin-10 were identified as 2 newly synthesized secreted proteins caused by substantial N incorporation. The inhibition of tissue inhibitor of metalloproteases-1 expression in MIA PaCa-2 cells by oxythiamine treatment was first demonstrated by 2D gel electrophoresis and further validated by Western blotting and enzyme linked immunosorbent assay analyses. CONCLUSIONS: Our method of labeling protein with N amino acids in conjunction with serum-free cell culture allows the identification of actively secreted proteins from pancreatic cancer cells and is a useful method for serum biomarker discovery. PMID- 19904224 TI - Highly active state of autoimmune pancreatitis with mikulicz disease. AB - OBJECTIVE: Patients with autoimmune pancreatitis (AIP) sometimes present with Mikulicz disease (MD); however, the clinical features regarding these AIP patients with MD have not yet been fully elucidated. Our aim is to study the clinical differences between AIP with and without MD. METHODS: Twenty-eight AIP patients were divided into 2 groups, one with MD and one without it. The following factors having a possible association with the presence or absence of MD were investigated: sex; serum IgG and IgG4 levels; the presence or absence of antinuclear autoantibodies, jaundice, diabetes mellitus, swollen duodenal papilla, diffuse pancreatic swelling, spontaneous remission, and relapse. RESULTS: The MD and non-MD groups consisted of 5 AIP and 23 AIP patients, respectively. The results of univariate analysis revealed that AIP patients presenting with MD were significantly associated with a younger onset, female predominance, high serum IgG4 titer, and diffuse pancreatic swelling (P < 0.05). In 4 of the MD patients, onset preceded pancreatitis. CONCLUSIONS: Autoimmune pancreatitis patients presenting with MD tended to have different clinical features from the non-MD AIP patients, such as having an earlier onset, female tendency, and diffuse pancreatic swelling with a high titer of serum IgG4. Autoimmune pancreatitis with MD tended to precede gastroenterological events. PMID- 19904225 TI - Total parenteral nutrition attenuates cerulein-induced pancreatitis in rats. AB - OBJECTIVES: Our aim was to determine if total parenteral nutrition (TPN)-induced pancreatic atrophy and Hsp70 expression attenuates cerulein-induced pancreatitis in rats. METHODS: Rats were randomized to a 7-day course of saline infusion plus a semipurified diet or TPN, with or without an intravenous cerulein injection or vehicle on day 7, and killed 1 or 6 hours after the injection. Based on a pilot study, 1 hour was the primary time point. Pancreatic atrophy was determined by mass, protein, and DNA contents. Pancreatic heat shock protein 70 (Hsp70) expression was measured by Western analysis. Histological examination of the pancreas assessed for edema, inflammation, vacuolization, and apoptosis. Serum amylase activity was measured using the Phadebas assay. Pancreatic trypsinogen activation was measured using a fluorometric substrate assay. RESULTS: The saline infused rats fed orally gained significantly more weight than TPN rats. The TPN decreased the pancreatic mass and protein content and the protein-DNA ratio and increased the pancreatic DNA content compared with the saline. The TPN increased the pancreatic Hsp70 expression by 91% compared with the saline. The TPN reduced the cerulein-induced pancreatic histological edema, the vacuolization, and the inflammation compared with the saline. The increase in the serum amylase level after cerulein injection was significantly attenuated, and trypsinogen activation was reduced in TPN animals compared with the saline group. CONCLUSIONS: Lack of luminal nutrients with a 7-day course of TPN provides moderate protection against cerulein-induced pancreatitis in rats. PMID- 19904232 TI - Extraction of high molecular weight genomic DNA from soils and sediments. AB - The soil microbiome is a vast and relatively unexplored reservoir of genomic diversity and metabolic innovation that is intimately associated with nutrient and energy flow within terrestrial ecosystems. Cultivation-independent environmental genomic, also known as metagenomic, approaches promise unprecedented access to this genetic information with respect to pathway reconstruction and functional screening for high value therapeutic and biomass conversion processes. However, the soil microbiome still remains a challenge largely due to the difficulty in obtaining high molecular weight of sufficient quality for large insert library production. Here we introduce a protocol for extracting high molecular weight, microbial community genomic DNA from soils and sediments. The quality of isolated genomic DNA is ideal for constructing large insert environmental genomic libraries for downstream sequencing and screening applications. The procedure starts with cell lysis. Cell walls and membranes of microbes are lysed by both mechanical (grinding) and chemical forces (beta mercaptoethanol). Genomic DNA is then isolated using extraction buffer, chloroform-isoamyl alcohol and isopropyl alcohol. The buffers employed for the lysis and extraction steps include guanidine isothiocyanate and hexadecyltrimethylammonium bromide (CTAB) to preserve the integrity of the high molecular weight genomic DNA. Depending on your downstream application, the isolated genomic DNA can be further purified using cesium chloride (CsCl) gradient ultracentrifugation, which reduces impurities including humic acids. The first procedure, extraction, takes approximately 8 hours, excluding DNA quantification step. The CsCl gradient ultracentrifugation, is a two days process. During the entire procedure, genomic DNA should be treated gently to prevent shearing, avoid severe vortexing, and repetitive harsh pipetting. PMID- 19904233 TI - Adipose-derived stem cells as therapeutic delivery vehicles of an oncolytic virus for glioblastoma. AB - Glioblastoma multiforme (GBM) accounts for the majority of primary malignant brain tumors and remains virtually incurable despite extensive surgical resection, radiotherapy, and chemotherapy. Treatment difficulty is due to its exceptional infiltrative nature and proclivity to integrate into normal brain tissue. Long-term survivors are rare, and median survival for patients is about 1 year. Use of adult stem cells as cellular delivery vehicles for anticancer agents is a novel attractive therapeutic strategy. We hypothesized that adipose-derived stem cells (ADSCs) possess the ability to home and deliver myxoma virus to glioma cells and experimental gliomas. We infected ADSCs with vMyxgfp and found them to be permissive for myxoma virus replication. ADSCs supported single and multiple rounds of replication leading to productive infection. Further, we observed no significant impact on ADSC viability. We cocultured fluorescently labeled GBM cells with myxoma virus-infected ADSCs in three-dimensional assay and observed successful cross infection and concomitant cell death almost exclusively in GBM cells. In vivo orthotopic studies injected with vMyxgfp-ADSCs intracranially away from the tumor demonstrated that myxoma virus was delivered by ADSCs resulting in significant survival increase. Our data suggest that ADSCs are promising new carriers of oncolytic viruses, specifically myxoma virus, to brain tumors. PMID- 19904234 TI - Effect of genome size on AAV vector packaging. AB - Adeno-associated virus (AAV) vector genomes have been limited to 5 kilobases (kb) in length because their packaging limit was thought to be similar to the size of the parent AAV genome. Recent reports claim that significantly larger vector genomes can be packaged intact. We examined the packaged vector genomes from plasmid-encoded AAV vectors that ranged from 4.7 to 8.7 kb in length, using AAV types 2, 5, and 8 capsids. Southern blot analysis indicated that packaged AAV vector genomes never exceeded 5.2 kb in length irrespective of the size of the plasmid-encoded vector or the capsid type. This result was confirmed by vector genome probing with strand-specific oligonucleotides. The packaged vector genomes derived from plasmid-encoded vectors exceeding 5 kb were heterogeneous in length and truncated on the 5' end. Despite their truncated genomes, vector preparations produced from plasmid-encoded vectors exceeding 5.2 kb mediated reporter gene expression in vitro at high multiplicity of infection (MOI). The efficiency of expression was substantially lower than that of reporter vectors with genomes <5 kb in length. We propose that transcriptionally functional, intact vector genomes are generated in cells transduced at high MOI from the fragmentary genomes of these larger vectors, probably by recombination. PMID- 19904235 TI - Proteasome inhibitors decrease AAV2 capsid derived peptide epitope presentation on MHC class I following transduction. AB - Adeno-associated viral (AAV) vectors are an extensively studied and highly used vector platform for gene therapy applications. We hypothesize that in the first clinical trial using AAV to treat hemophilia B, AAV capsid proteins were presented on the surface of transduced hepatocytes, resulting in clearance by antigen-specific CD8+ T cells and consequent loss of therapeutic transgene expression. It has been previously shown that proteasome inhibitors can have a dramatic effect on AAV transduction in vitro and in vivo. Here, we describe using the US Food and Drug Administration-approved proteasome inhibitor, bortezomib, to decrease capsid antigen presentation on hepatocytes in vitro, whereas at the same time, enhancing gene expression in vivo. Using an AAV capsid-specific T-cell reporter (TCR) line to analyze the effect of proteasome inhibitors on antigen presentation, we demonstrate capsid antigen presentation at low multiplicities of infection (MOIs), and inhibition of antigen presentation at pharmacologic levels of bortezomib. We also demonstrate that bortezomib can enhance Factor IX (FIX) expression from an AAV2 vector in mice, although the same effect was not observed for AAV8 vectors. A pharmacological agent that can enhance AAV transduction, decrease T-cell activation/proliferation, and decrease capsid antigen presentation would be a promising solution to obstacles to successful AAV mediated, liver-directed gene transfer in humans. PMID- 19904236 TI - Characterization of genome integrity for oversized recombinant AAV vector. AB - Application of recombinant adeno-associated virus (rAAV) in gene therapy has been limited by its packaging capacity. Recent studies suggested that rAAV could achieve persistent transgene expression beyond 4.7-kb packaging limit. To clarify the mechanism leading to transgene expression from oversized rAAV vector, we constructed a series of rAAV vectors with genomes ranging from 2.9 to 7.2 kb. A plasmid replication origin and an ampicillin-resistant marker were included in the vector to facilitate the recovery of circularized, post-transduction AAV genome. Southern dot-blot analysis and silver staining confirmed that rAAVs could be produced at varying vector size. However, the vector yields decreased approximately tenfold for oversized vectors as compared to regular ones. Alkaline Southern blot hybridization suggested that the packaged genomes for oversized vectors were truncated. In the cells transduced by the above vectors, circularized rAAV monomers could be rescued at 24 hours after infection. Few recovered AAV genomes were >5 kb regardless of the initial vector size. In mice receiving the above vectors, larger circularized rAAV genomes could be recovered for oversized vectors at day 21 after vector administration. Our studies suggested that the partially packaged rAAV sequences may complement each other to restore full expression cassette. PMID- 19904237 TI - Persistent expression of FLAG-tagged micro dystrophin in nonhuman primates following intramuscular and vascular delivery. AB - Animal models for Duchenne muscular dystrophy (DMD) have species limitations related to assessing function, immune response, and distribution of micro- or mini-dystrophins. Nonhuman primates (NHPs) provide the ideal model to optimize vector delivery across a vascular barrier and provide accurate dose estimates for widespread transduction. To address vascular delivery and dosing in rhesus macaques, we have generated a fusion construct that encodes an eight amino-acid FLAG epitope at the C-terminus of micro-dystrophin to facilitate translational studies targeting DMD. Intramuscular (IM) injection of AAV8.MCK.micro-dys.FLAG in the tibialis anterior (TA) of macaques demonstrated robust gene expression, with muscle transduction (50-79%) persisting for up to 5 months. Success by IM injection was followed by targeted vascular delivery studies using a fluoroscopy guided catheter threaded through the femoral artery. Three months after gene transfer, >80% of muscle fibers showed gene expression in the targeted muscle. No cellular immune response to AAV8 capsid, micro-dystrophin, or the FLAG tag was detected by interferon-gamma (IFN-gamma) enzyme-linked immunosorbent spot (ELISpot) at any time point with either route. In summary, an epitope-tagged micro-dystrophin cassette enhances the ability to evaluate site-specific localization and distribution of gene expression in the NHP in preparation for vascular delivery clinical trials. PMID- 19904238 TI - Evidence for the failure of adeno-associated virus serotype 5 to package a viral genome > or = 8.2 kb. AB - Limited packaging capacity hinders adeno-associated virus (AAV) gene therapy. A recent study seems to have provided a solution to this problem. Allocca et al. reported that AAV-5 could package an 8.9 kb vector genome. Here we tested whether this approach can be used to deliver a large genome for Duchenne muscular dystrophy (DMD) gene therapy. We first evaluated AAV-5 packaging of an 8.2 kb genome. This vector carries two independent reporter gene cassettes, one for alkaline phosphatase (AP) and another for LacZ. Viral yield was log-fold lower than that of a regular AAV-5. Nevertheless, both AP and LacZ genes were detected in purified virus. Injection to dystrophic muscle resulted in both AP and LacZ expression. On electron microscopy, virion structure appeared normal. Surprisingly, we did not find the full-length single-stranded viral genome by alkaline gel electrophoresis. Neither did we see the full-length double-stranded replication forms in adenovirus coinfected cells. We suspect that AP and LacZ expression may have come from partially packaged 5' or 3'-half of the genome. Additional studies revealed failure of AAV-5 to package and express an 8.7 kb minidystrophin gene cassette. In summary, our results do not support the extraordinary packaging capacity of AAV-5. PMID- 19904239 TI - Predictors of colorectal cancer after negative colonoscopy: a population-based study. AB - OBJECTIVES: A higher proportion of colorectal neoplasia among women occurs in the proximal colon, which might be more frequently missed by colonoscopy. There are no data on predictors of developing colorectal cancer (CRC) after a negative colonoscopy in usual clinical practice. We evaluated gender differences and predictors of CRC occurring after a negative colonoscopy. METHODS: All individuals 40 years or older with negative colonoscopy were identified from Manitoba's provincial physicians' billing claims database. Individuals with less than 5 years of coverage by the provincial health plan, earlier CRC, inflammatory bowel disease, resective colorectal surgery, or lower gastrointestinal endoscopy were excluded. CRC risk after negative colonoscopy was compared to that in the general population by standardized incidence ratios. Cox regression analysis was performed to determine the independent predictors of CRC occurring after negative colonoscopy. RESULTS: A total of 45,985 individuals (18,606 men; 27,379 women) were followed up for 229,090 person-years. After a negative colonoscopy, men had a 40-50% lower risk of CRC diagnosis through most of the follow-up time. Risk among women was similar to that of women in the general population in the first 3 years and then was 40-50% lower. Older subject age and performance of index colonoscopy by non-gastroenterologists were independent predictors for early/missed CRC (cancers occurring within 3 years of negative colonoscopy). CONCLUSIONS: Women may have a higher rate of missed/early CRCs after negative colonoscopy. Predictors of missed/early CRCs after negative colonoscopy include older age and performance of index colonoscopy by a non-gastroenterologist. PMID- 19904240 TI - IL-1B-511 polymorphism is associated with increased risk of certain subtypes of gastric cancer in Chinese: a case-control study. AB - OBJECTIVES: The association of interleukin-1B (IL-1B)-511 polymorphism with gastric cancer is still controversial, and the association of IL-1B-511 polymorphism with subtypes of gastric cancer is still largely unknown. We investigated whether the association between IL-1B-511 polymorphism and gastric cancer risk varies by clinically important tumor characteristics and the prognostic value of this polymorphism in a large population-based case-control study among Chinese. METHODS: A population-based case-control study was conducted between 1999 and 2006 in Guangdong Province, China. A total of 1,010 gastric cancer patients and 1,500 healthy controls were enrolled in this study. Polymorphism in IL-1B-511 was analyzed by PCR-restriction fragment length polymorphism on 501 gastric cancers and 500 healthy controls. RESULTS: Compared with the CC genotype, carriers of IL-1B-511 TT genotype had an increased gastric cancer risk (odds ratio (OR)=1.97, 95% confidence interval (CI)=1.29-3.01, P=0.0016). TT genotype was significantly associated with intestinal type of gastric cancer (OR=3.16, 95% CI=1.74-5.71, P=0.0001) but not with diffuse or mixed-type gastric cancer. The test for OR heterogeneity between the intestinal type and non-intestinal-type gastric cancers was statistically significant (P=0.02). In subgroup analyses, TT genotype was found to be associated with poorly differentiated gastric cancer (OR=3.31, 95% CI=1.43-3.60, P<0.0001), but not with moderately or well-differentiated gastric cancer. IL-1B-511 genotypes were not associated with the prognosis of gastric cancer patients. CONCLUSIONS: IL-1B polymorphism influences certain subtypes of gastric cancer according to the clinical and pathological features. Understanding the etiologic heterogeneity of gastric cancer may result in improvements in controlling this disorder. PMID- 19904241 TI - Proton pump inhibitor and clopidogrel interaction: fact or fiction? AB - Current consensus recommendations state that patients prescribed clopidogrel plus aspirin should receive a proton pump inhibitor (PPI) to reduce gastrointestinal bleeding. Clopidogrel is converted to its active metabolite by cytochrome P450 (CYP) enzymes. Clopidogrel users with decreased CYP2C19 function have less inhibition of platelet aggregation and increased cardiovascular (CV) events. As PPI metabolism also involves CYP2C19, it was hypothesized that competition by PPIs might interfere with clopidogrel's action. Omeprazole, but not other PPIs, worsens surrogate markers of clopidogrel efficacy. Some (but not all) observational studies show that clopidogrel users prescribed PPIs have increased risks of CV events (hazard/odds ratios=1.25-1.5). When effect sizes are small to moderate (relative risks<1.5-2.0), however, it is only possible to conclude whether statistical associations are valid in randomized trials. A randomized trial of omeprazole vs. placebo in clopidogrel users showed no difference in CV events (hazard ratio=1.02,0.70-1.51). Thus, current evidence does not justify a conclusion that PPIs are associated with CV events among clopidogrel users, let alone a judgment of causality. Nonetheless, positive results from some observational studies and biological plausibility have led some health-care providers to accept that PPIs reduce clopidogrel's efficacy. The US Food and Drug Administration (FDA) recommends that "concomitant use of drugs that inhibit CYP2C19 (e.g., omeprazole) should be discouraged." As the presence of PPIs and clopidogrel in plasma is short lived, separation by 12-20 h should in theory prevent competitive inhibition of CYP metabolism and minimize any potential, though unproven, clinical interaction. PPI may be given before breakfast and clopidogrel at bedtime, or PPI may be taken before dinner and clopidogrel at lunchtime. PMID- 19904242 TI - Reassessment of the principal characteristics of gastroesophageal reflux during the recumbent period using integrated actigraphy-acquired information. AB - OBJECTIVES: Characterization of gastroesophageal reflux (GERD) events during the sleep period has been hampered by lack of any patient-friendly technique that allows accurate assessment of sleep duration and awakening time, without confining patients to a sleep laboratory. Our aim was to compare principal reflux characteristics during the upright, recumbent-awake, and recumbent-asleep periods as well as to determine the effect of sleep awakenings on the principal reflux characteristics of the recumbent-asleep period using novel technology that allows integration of recorded actigraphy data into collected pH information. METHODS: Patients with heartburn at least three times a week for the previous 3 months were invited to participate in this study. All participants were evaluated by the demographics and the GERD Symptom Checklist questionnaires. Thereafter, patients underwent ambulatory 24-h esophageal pH monitoring concomitantly with actigraphy. A novel technique was used to superimpose simultaneously recorded raw actigraphy data over pH data, resulting in more accurate information about reflux events during upright, recumbent-awake, recumbent-asleep, and conscious awakening periods as well as the relationship between symptoms and acid reflux events in the aforementioned periods. RESULTS: Thirty-nine subjects (M/F: 26/13, mean age 56.6+/-14 years) with an abnormal pH test were enrolled into the study. The recumbent period appeared heterogeneous and was clearly divided into recumbent awake (123.0+/-20.2 min) and recumbent-asleep (485.6+/-23.6 min) periods. The percent total time pH<4, the mean number of acid reflux events, and the number of symptoms associated with reflux events were significantly greater in the recumbent-awake as compared with the recumbent-asleep period. The mean duration of an acid reflux event was not different among upright, recumbent-awake, and recumbent-asleep periods. However, short-duration reflux events during the sleep period were associated with conscious awakenings as compared with those during sleep (0.74+/-0.11 min vs. 1.64+/-0.3 min, P=0.01). CONCLUSIONS: The recumbent period is divided into recumbent-awake and recumbent-asleep periods. The recumbent-awake period has significantly different principal reflux characteristics than the recumbent-asleep period. Duration of an acid reflux event during the recumbent-asleep period is not uniformly prolonged. Short duration acid reflux events during the sleep period are likely due to conscious awakenings. PMID- 19904243 TI - Defective paneth cell-mediated host defense in pediatric ileal Crohn's disease. AB - OBJECTIVES: Adult ileal Crohn's disease (CD) is characterized by a specific decrease in ileal Paneth cell alpha-defensins. In addition to NOD2, we previously identified a disturbance of the Wnt-signaling transcription factor TCF-4 as a major mechanism for this deficiency. The aim of this study was to evaluate human alpha-defensin-5 (HD-5) and TCF-4 in an independent cohort of pediatric CD patients. METHODS: Expression levels of HD-5 and TCF-4 mRNA were quantified by real-time PCR in biopsies from newly diagnosed untreated pediatric CD patients (<18 years, n=36) and age-matched symptomatic non-inflammatory bowel disease controls with a histologically normal gut (n=29). To assess the influence of current inflammation, mucosal interleukin-8 (IL-8) and fecal calprotectin levels were determined. RESULTS: Small intestinal HD-5 and TCF-4 mRNA were significantly reduced in pediatric ileal CD (L1+L3) (P=0.022 and P=0.0005, respectively) and were significantly correlated (r=0.499; P=0.0001). In ileal but not colonic CD, TCF-4 was also reduced in the colon (P=0.005). Importantly, both HD-5 and TCF-4 were independent of inflammation, as measured by IL-8 expression or fecal calprotectin. In contrast to the small intestine, colonic Paneth cell HD-5 mRNA was significantly elevated in colonic CD (L2) (P=0.026) and was correlated with fecal calprotectin levels (r=0.481; P=0.020). CONCLUSIONS: In this study, we describe a specific decrease in HD-5 and TCF-4 mRNA expression levels in children with ileal CD. In the small intestine, this decrease was independent of current inflammation, whereas inflammation seems to induce Paneth cell metaplasia in the colon. Our data extend the hypothesis of an important role of antimicrobial host defense in pediatric CD patients. PMID- 19904244 TI - Anti-pituitary antibodies in children with newly diagnosed celiac disease: a novel finding contributing to linear-growth impairment. AB - OBJECTIVES: The possible autoimmune involvement of the pituitary gland in patients with celiac disease (CD) has been suggested but demonstrated in only a few patients on gluten-free diet. We aimed to assess the prevalence and clinical meaning of anti-pituitary antibodies (APA) in children and adolescents with the newly diagnosed CD. METHODS: A total of 119 patients with CD (0.9-15.8 years old) attending the inpatient clinic of University Hospital were recruited for the cross-sectional study. Their height, weight, and body mass index (BMI) were recorded, and insulin-like growth factor-1 (IGF-1) and APA were assayed. APA was also determined in 98 sex- and age-matched controls. RESULTS: APA were detected in 50 patients (42.0%), 15 of them with high titer (30%) and 35 with low titer (70%), and in 2 control subjects at low titer (2%) (P<0.001). IGF-1 was higher in patients with negative than with low titer (P=0.02) or high titer APA (P=0.03). Height was more reduced in high-titer APA patients than in the negative ones (P<0.01). Height was positively correlated with IGF-1 (P<0.01) and negatively with chronological age (P=0.001). IGF-1 was positively correlated with BMI (P<0.001). For height prediction the regression analysis showed the rank order 1 for chronological age and 2 for IGF-1. CONCLUSIONS: In this paper we have shown a remarkable prevalence of positive APA in newly diagnosed CD patients. High APA titers are associated with height impairment, likely mediated by a reduction of IGF-1, thus suggesting that autoimmune pituitary process could induce a linear growth impairment. PMID- 19904245 TI - Transjugular intrahepatic portosystemic shunt for symptomatic refractory hepatic hydrothorax in patients with cirrhosis. AB - OBJECTIVES: We sought to study effectiveness, survival, and complications after transjugular intrahepatic portosystemic shunt (TIPS) in patients with cirrhosis and symptomatic refractory hepatic hydrothorax. METHODS: Consecutive patients who underwent TIPS between January 1992 and December 2008 for refractory hydrothorax were reviewed retrospectively. Clinical, laboratory, and procedural data were collected for all patients by retrospective chart review. Chi-square test was used to compare categorical variables and t-test to compare continuous variables. The Kaplan-Meier method was used for survival analysis. Survival curves were compared using the log-rank test. RESULTS: Seventy-three patients were included in the study, and their mean age at TIPS creation was 55.62 years (s.d. 11.65). The mean pre- and post-TIPS portosystemic gradients were 18.9 (s.d. 4.7) mm Hg and 5.7 (s.d. 2.4) mm Hg (P<0.001), respectively. The rates of favorable clinical response within 1 month and at 6 months after TIPS were 79% (58/73) and 75% (30/40), respectively. Median survival of the study group was 517 days (95% CI 11 626). The short-term survival rates at 30, 60, and 90 days were 81, 78, and 72%, respectively. The long-term survival rates at 1, 3, and 5 years were 48, 26, and 15%, respectively. Multivariate analysis by Cox proportional hazards method showed that pre-TIPS model for end-stage liver disease (MELD) score (P=0.039, HR 1.9 (95% CI 1.0-3.7)) and clinical response (P=0.003, HR 2.5 (95% CI 1.4-4.5)) were significantly and independently associated with overall survival. The 30-day mortality rate was 19%. Pre-TIPS creatinine levels (P=0.024, HR 3.42 (95% CI 1.2 9.9)) were significantly associated with 30-day mortality. CONCLUSIONS: TIPS can be successfully used to achieve symptomatic relief in patients with refractory hepatic hydrothorax. Better clinical response after TIPS and pre-TIPS MELD score less than 15 were associated with longer survival after TIPS. PMID- 19904247 TI - Similar treatment response to peginterferon and ribavirin in Asian and Caucasian patients with chronic hepatitis C. AB - OBJECTIVES: Previous studies have found ethnicity to be an important predictor of outcomes of treatment with peginterferon (PEG-IFN) and ribavirin (RBV) in chronic hepatitis C. Although the expected sustained virological response (SVR) rates of Hispanics and African Americans are lower than those of Caucasians, SVR rates in Asians appear to be more favorable. However, in some of these studies, hepatitis C virus (HCV) genotype was identified by INNO-LiPA assay, which can mistype the easier-to-treat HCV genotype 6 as genotype 1. Our goal was to compare SVR rates among Caucasian and Asian-American patients with genotype 1 and 2/3 infection whose HCV genotypes were accurately classified by core sequencing testing. METHODS: A cohort of 269 consecutive treatment-naive HCV-infected patients with genotype 1 or 2/3 (157 Caucasians and 112 Asians) treated with PEG-IFN+RBV from January 2001 to November 2007 at four community-based gastroenterology clinics in Northern California were studied. The analysis of data was by intention-to-treat. RESULTS: The SVR rates for patients with genotype 1 were 45% for Caucasians and 52% for Asians (P=0.37). The SVR rates for patients with genotype 2/3 infection was 77% for Asians and 74% for Caucasians (P=0.7). On multivariate logistic regression analyses adjusting for age, alanine aminotransferase (ALT), baseline viral load, HCV genotype, and treatment adherence, we did not find Asian ethnicity to predict SVR. On a separate analysis, we found that Asians who had HCV genotype 1 or 1b by the less accurate INNO-LiPA assay had significantly higher SVR rates than Caucasians with genotype 1 (64% vs. 45%, respectively, P=0.03). CONCLUSIONS: SVR rates were similar in Asian Americans and Caucasians infected with HCV genotype 1 or 2/3 when HCV genotype classification was accurately determined. PMID- 19904248 TI - Alcoholic liver disease. AB - These recommendations provide a data-supported approach. They are based on the following: (i) a formal review and analysis of the recently published world literature on the topic (Medline search); (ii) American College of Physicians Manual for Assessing Health Practices and Designing Practice Guidelines (1); (iii) guideline policies, including the American Association for the Study of Liver Diseases (AASLD) Policy on the development and use of practice guidelines and the AGA Policy Statement on Guidelines (2); and (iv) the experience of the authors in the specified topic. Intended for use by physicians, these recommendations suggest preferred approaches to the diagnostic, therapeutic, and preventive aspects of care. They are intended to be flexible, in contrast to the standards of care, which are inflexible policies to be followed in every case. Specific recommendations are based on relevant published information. To more fully characterize the quality of evidence supporting the recommendations, the Practice Guideline Committee of the AASLD requires a Class (reflecting the benefit vs. risk) and Level (assessing the strength or certainty) of Evidence to be assigned and reported with each recommendation (Table 1, adapted from the American College of Cardiology and the American Heart Association Practice Guidelines) (3,4). PMID- 19904246 TI - Long-term outcome of microscopic esophagitis in chronic GERD patients treated with esomeprazole or laparoscopic antireflux surgery in the LOTUS trial. AB - OBJECTIVES: Gastroesophageal reflux disease (GERD)-associated changes in esophageal histology have been reported mainly after short-term medical antireflux therapy, and few individual lesions have been examined. We report detailed histological findings from the LOTUS study, at baseline and at 1 and 3 years after laparoscopic antireflux surgery (LARS) or esomeprazole treatment in patients with chronic GERD. METHODS: LOTUS is a long-term, open, parallel-group, multicenter, randomized, controlled trial conducted in 11 European countries that compared LARS (n=248) with esomeprazole 20-40 mg daily (n=266). Biopsies from the distal esophagus 2 cm above the Z-line and at the Z-line were taken at baseline, and 1 and 3 years. The following lesions were assessed: basal cell hyperplasia (BCH), papillary elongation (PE), intercellular space dilatations (ISDs), intraepithelial eosinophils (EOSs), neutrophils, and necrosis/erosion. A severity score (SS, range 0-2) was calculated by taking the average score of all assessable lesions. RESULTS: All lesions were more severe on Z-line biopsies than at 2 cm, and almost all improved significantly from baseline to 1 and 3 years. The average SS (from 2 cm to Z-line) changed from 0.95 to 0.57 (1 year) and to 0.49 (3 years) on esomeprazole, and from 0.91 to 0.56 (1 year) and to 0.52 (3 years) after LARS (P<0.001 for both treatments at 1 and 3 years, with no significant difference between treatments). The proportions of patients with severe histological changes decreased from approximately 50% at baseline to 11% at 3 years. CONCLUSIONS: Both continuous esomeprazole treatment and laparoscopic fundoplication are associated with significant and similar overall improvement in microscopic esophagitis after 1 year that is maintained at 3 years. PMID- 19904249 TI - Longitudinal profiles of 15 serum bile acids in patients with intrahepatic cholestasis of pregnancy. AB - OBJECTIVES: Increased maternal serum bile acids are implicated in intrahepatic cholestasis of pregnancy. Individual bile acid profiles and their relationship with disease progression, however, remain unknown. The purpose of this prospective study was to determine the temporal changes in bile acids in normal pregnancy and in pregnancies complicated with intrahepatic cholestasis of pregnancy and pruritus gravidarum. METHODS: A validated method for the evaluation of 15 bile acids (conjugated and unconjugated) in a single serum sample was developed using high-performance liquid chromatography/mass spectrometry (HPLC MS) with an electrospray interface. Bile acid concentrations were assessed in samples (16 weeks of gestation to 4 weeks postpartum) from women with, or who later developed, intrahepatic cholestasis of pregnancy (n=63) and were compared with those from normal pregnant women (n=26) and from women with pruritus gravidarum (n=43). RESULTS: Intrahepatic cholestasis of pregnancy was associated with a predominant increase in cholic acid conjugated with taurine and glycine, from 24 weeks of pregnancy. Ursodeoxycholic acid (UDCA) treatment (> or =21 days, n=15) significantly reduced serum taurocholic and taurodeoxycholic acid concentrations (P<0.01). Bile acid profiles were similar in normal pregnancy and pregnancy associated with pruritus gravidarum. CONCLUSIONS: The bile acid profiles and effects of treatment by UDCA implicate a role for taurine-conjugated bile acids in the syndrome of intrahepatic cholestasis of pregnancy. [corrected] With regard to individual bile acid profiles, pruritus gravidarum is a disorder quite distinct from intrahepatic cholestasis of pregnancy. PMID- 19904250 TI - Double-dosed pantoprazole accelerates the sustained symptomatic response in overweight and obese patients with reflux esophagitis in Los Angeles grades A and B. AB - OBJECTIVES: Body mass index (BMI) in the range defined as overweight or obese adversely decreases the sustained symptomatic response (SSR) to proton pump inhibitors for patients with reflux esophagitis of Los Angeles grade A or B (RE AB). We thus investigated whether double-dosed pantoprazole can accelerate SSR in such patients. METHODS: A total of 200 overweight or obese patients with RE-AB were evenly randomized into a double-dosed group (receiving 8-week pantoprazole 40 mg twice daily) or a standard-dosed control group (receiving 8-week pantoprazole 40 mg per day and one blank tablet at night). In each patient, demographic factors and the genotype of S-mephenytoin 4'-hydroxylase (CYP2C19) were checked and defined as poor metabolizer (PM), or homologous extensive metabolizer (HomoEM), or heterologous extensive metabolizer (HeteroEM). The cumulative proportions of patients with SSR were compared during the 8-week period. RESULTS: Both intention-to-treat and per-protocol analyses disclosed that the rates of SSR were higher in the double-dosed group than in the standard-dosed group from week 4 (P=0.005) until week 8 (P=0.01). While using standard-dosed pantoprazole, PMs had better rates of SSR during the 8-week period than both HomoEMs and HeteroEMs (P<0.05). By using double-dosed pantoprazole, the cumulative rates of SSR were improved as early as week 4 for both HomoEMs and HeteroEMs (P<0.005, log-rank test). CONCLUSIONS: For RE-AB in overweight and obese patients, double-dosed pantoprazole effectively accelerates the SSR, especially for those with CYP2C19 genotypes as HeteroEM or HomoEM. Accordingly, it offers an earlier shift into on-demand pantoprazole for RE-AB patients with high BMI. PMID- 19904253 TI - The International Society of Nephrology: what does "philanthropic" mean for members? PMID- 19904251 TI - Elevated serum gastrin is associated with a history of advanced neoplasia in Barrett's esophagus. AB - OBJECTIVES: Proton pump inhibitors (PPIs) are frequently prescribed to patients with Barrett's esophagus (BE), but in a subset, they can induce significant hypergastrinemia. Elevated levels of gastrin have been associated with tumorigenic effects in a number of gastrointestinal cancers. We decided to investigate the association between serum gastrin levels and dysplasia in BE. METHODS: We performed a cross-sectional study and enrolled patients with BE without dysplasia, low-grade dysplasia (LGD), high-grade dysplasia (HGD), or adenocarcinoma (AC), as well as gastroesophageal reflux disease controls, all chronically taking PPIs. Fasting serum gastrin was measured, and data were collected on patient characteristics, medication use, and the highest degree of BE neoplasia. RESULTS: A total of 95 patients were enrolled. The mean age was 64.7 (+/-10.0) years, and 70.5% were male. The median serum gastrin level was 40 pM. There was no significant difference in gastrin levels with increased degrees of BE neoplasia (overall P=0.68). In multivariable analysis, the highest quartile of gastrin was associated with significantly increased odds of advanced neoplasia (HGD or AC) (odds ratio (OR): 5.46, 95% confidence interval (CI): 1.20-24.8). CONCLUSIONS: In BE patients taking PPIs, an elevated serum gastrin is associated with a history of HGD or AC. Prospective studies are needed to determine whether patients with nondysplastic BE and elevated serum gastrin are at increased risk for neoplastic progression. PMID- 19904255 TI - Impact of reporting estimated glomerular filtration rate: it's not just about us. AB - The reporting of estimated glomerular filtration rate (eGFR) is a key component of a public-health strategy for chronic kidney disease (CKD). Jain and colleagues show that eGFR reporting in Ontario increased referrals to nephrologists by 23 consults per year, equivalent to 2.9 additional consults per 100,000 population. In our view, a complete assessment requires a broader perspective. Most patients with CKD do not require referral to a nephrologist. Improving outcomes requires coordinated efforts of all physicians. PMID- 19904256 TI - Atrial fibrillation in end-stage renal disease: an emerging problem. AB - End-stage renal disease is associated with considerable cardiovascular morbidity and mortality. Atrial fibrillation (AF) is prevalent among dialysis patients and adversely affects the clinical outcome. Vazquez et al. report an increased prevalence and incidence of AF in patients who commence dialysis, and an independent association between arrhythmia and mortality risk. Moreover, they examined clinical and laboratory parameters associated with AF. This population study may fuel research aiming at the development of novel preventive and treatment strategies. PMID- 19904257 TI - Preventing repeat hospitalizations in dialysis patients: a call for action. AB - Hospitalizations are frequent among dialysis patients, and reducing repeat hospitalizations could decrease costs and improve outcomes. Chan et al. found that hemoglobin monitoring along with erythropoietin-stimulating agent dose modification and vitamin D administration reduced the risk of repeat hospitalization. These and other interventions, especially those related to close monitoring immediately after hospitalization and better communication between hospital and dialysis providers to assure continuity of care, should be further explored in observational or randomized studies. PMID- 19904259 TI - The case: a female with hyponatremia. Diagnosis: Postpartum panhypopituitarism (Sheehan syndrome). PMID- 19904260 TI - Detection of unrecognized urinothorax with renal scintigraphy. PMID- 19904261 TI - Dissociable cost and benefit encoding of future rewards by mesolimbic dopamine. AB - Reward-predicting cues evoke activity in midbrain dopamine neurons that encodes fundamental attributes of economic value, including reward magnitude, delay and uncertainty. We found that dopamine release in rat nucleus accumbens encodes anticipated benefits, but not effort-based response costs unless they are atypically low. This neural separation of costs and benefits indicates that mesolimbic dopamine scales with the value of pending rewards, but does not encode the net utility of the action to obtain them. PMID- 19904262 TI - Self-production of tissue factor-coagulation factor VII complex by ovarian cancer cells. AB - BACKGROUND: Thromboembolic events are a major complication in ovarian cancer patients. Tissue factor (TF) is frequently overexpressed in ovarian cancer tissue and correlates with intravascular thrombosis. TF binds to coagulation factor VII (fVII), changing it to its active form, fVIIa. This leads to activation of the extrinsic coagulation cascade. fVII is produced by the liver and believed to be supplied from blood plasma at the site of coagulation. However, we recently showed that ovarian cancer cells express fVII transcripts under normoxia and that this transcription is inducible under hypoxia. These findings led us to hypothesise that ovarian cancer cells are intrinsically associated with TF-fVIIa coagulation activity, which could result in thrombosis. METHODS: In this study, we examined whether ectopically expressed fVII could cause thrombosis by means of immunohistochemistry, RT-PCR, western blotting and flow cytometry. RESULTS: Ectopic fVII expression occurs frequently in ovarian cancers, particularly in clear cell carcinoma. We further showed that ovarian cancer cells express TF fVIIa on the cell surface under normoxia and that this procoagulant activity is enhanced by hypoxic stimuli. Moreover, we showed that ovarian cancer cells secrete microparticles (MPs) with TF-fVIIa activity. Production of this procoagulant secretion is enhanced under hypoxia. CONCLUSION: These results raise the possibility that cancer cell-derived TF-fVIIa could cause thrombotic events in ovarian cancer patients. PMID- 19904263 TI - Multicentre phase II studies evaluating imatinib plus hydroxyurea in patients with progressive glioblastoma. AB - BACKGROUND: We evaluated the efficacy of imatinib mesylate in addition to hydroxyurea in patients with recurrent glioblastoma (GBM) who were either on or not on enzyme-inducing anti-epileptic drugs (EIAEDs). METHODS: A total of 231 patients with GBM at first recurrence from 21 institutions in 10 countries were enrolled. All patients received 500 mg of hydroxyurea twice a day. Imatinib was administered at 600 mg per day for patients not on EIAEDs and at 500 mg twice a day if on EIAEDs. The primary end point was radiographic response rate and secondary end points were safety, progression-free survival at 6 months (PFS-6), and overall survival (OS). RESULTS: The radiographic response rate after centralised review was 3.4%. Progression-free survival at 6 months and median OS were 10.6% and 26.0 weeks, respectively. Outcome did not appear to differ based on EIAED status. The most common grade 3 or greater adverse events were fatigue (7%), neutropaenia (7%), and thrombocytopaenia (7%). CONCLUSIONS: Imatinib in addition to hydroxyurea was well tolerated among patients with recurrent GBM but did not show clinically meaningful anti-tumour activity. PMID- 19904264 TI - Trends in non-metastatic prostate cancer management in the Northern and Yorkshire region of England, 2000-2006. AB - BACKGROUND: Our objective was to analyse variation in non-metastatic prostate cancer management in the Northern and Yorkshire region of England. METHODS: We included 21,334 men aged > or = 55, diagnosed between 2000 and 2006. Principal treatment received was categorised into radical prostatectomy (11%), brachytherapy (2%), external beam radiotherapy (16%), hormone therapy (42%) and no treatment (29%). RESULTS: The odds ratio (OR) for receiving a radical prostatectomy was 1.53 in 2006 compared with 2000 (95% CI 1.26-1.86), whereas the OR for receiving hormone therapy was 0.57 (0.51-0.64). Age was strongly associated with treatment received; radical treatments were significantly less likely in men aged > or = 75 compared with men aged 55-64 years, whereas the odds of receiving hormone therapy or no treatment were significantly higher in the older age group. The OR for receiving radical prostatectomy, brachytherapy or external beam radiotherapy were all significantly lower in the most deprived areas when compared with the most affluent (0.64 (0.55-0.75), 0.32 (0.22-0.47) and 0.83 (0.74-0.94), respectively) whereas the OR for receiving hormone therapy was 1.56 (1.42-1.71). CONCLUSIONS: This study highlights the variation and inequalities that exist in the management of non-metastatic prostate cancer in the Northern and Yorkshire region of England. PMID- 19904265 TI - Identification of TNF-alpha and MMP-9 as potential baseline predictive serum markers of sunitinib activity in patients with renal cell carcinoma using a human cytokine array. AB - BACKGROUND: Several drugs are available to treat metastatic renal-cell carcinoma (MRCC), and predictive markers to identify the most adequate treatment for each patient are needed. Our objective was to identify potential predictive markers of sunitinib activity in MRCC. METHODS: We collected sequential serum samples from 31 patients treated with sunitinib. Sera of six patients with extreme phenotypes of either marked responses or clear progressions were analysed with a Human Cytokine Array which evaluates 174 cytokines before and after treatment. Variations in cytokine signal intensity were compared between both groups and the most relevant cytokines were assessed by ELISA in all the patients. RESULTS: Twenty-seven of the 174 cytokines varied significantly between both groups. Five of them (TNF-alpha, MMP-9, ICAM-1, BDNF and SDF-1) were assessed by ELISA in 21 evaluable patients. TNF-alpha and MMP-9 baseline levels were significantly increased in non-responders and significantly associated with reduced overall survival and time-to-progression, respectively. The area under the ROC curves for TNF-alpha and MMP-9 as predictive markers of sunitinib activity were 0.83 and 0.77. CONCLUSION: Baseline levels of TNF-alpha and MMP-9 warrant further study as predictive markers of sunitinib activity in MRCC. Selection of patients with extreme phenotypes seems a valid method to identify potential predictive factors of response. PMID- 19904266 TI - Vitamin D receptor polymorphisms and prognosis of patients with epithelial ovarian cancer. AB - BACKGROUND: Recently, the vitamin D receptor (VDR) polymorphism FokI was shown to be associated with susceptibility to ovarian cancer. We aimed to examine whether VDR FokI polymorphisms influence the survivals of patients with epithelial ovarian cancer (EOC). METHODS: VDR polymorphisms from FokI in 101 patients with EOC were genotyped by sequencing. Overall survival was compared between FokI single nucleotide polymorphism using Kaplan-Meier survival curves with log-rank tests and the Cox proportional hazard model adjusted for ages, stages, histology, and existence of residual tumour. RESULTS: The FokI C/C genotypes were associated with better prognosis compared with the C/T and T/T genotypes (log-rank test: P = 0.008; adjusted hazard ratio, 0.18; 95%CI 0.05-0.61; P = 0.006). CONCLUSIONS: These results suggest that the VDR polymorphisms from the FokI genotype may be associated with improved prognosis of patients with EOC. PMID- 19904267 TI - Gemcitabine, oxaliplatin and 5-FU in advanced bile duct and gallbladder carcinoma: two parallel, multicentre phase-II trials. AB - BACKGROUND: Gemcitabine, oxaliplatin and 5-fluorouracil (5-FU) are active in biliary tract cancer and have a potentially synergistic mode of action and non overlapping toxicity. The objective of these trials was to determine response, survival and toxicity separately in patients with bile duct cancer (BDC) and gallbladder cancer (GBC) treated with gemcitabine/oxaliplatin/5-FU chemotherapy. METHODS: Eligible patients with histologically proven, advanced or metastatic BDC (n=37) or GBC (n=35) were treated with gemcitabine (900 mg m(-2) over 30 min), oxaliplatin (65 mg m(-2)) and 5-FU (1500 mg m(-2) over 24 h) on days 1 and 8 of a 21-day cycle. Tumour response was the primary outcome measure. RESULTS: Response rates were 19% (95% CI: 6-32%) and 23% (95% CI: 9-37%) for BDC and GBC, respectively. Median survivals were 10.0 months (95% CI: 8.6-12.4) and 9.9 months (95% CI: 7.5-12.2) for BDC and GBC, respectively, and 1- and 2-year survival rates were 40 and 23% in BDC and 34 and 6% in GBC (intention-to-treat analysis). Major grade III and IV adverse events were neutropenia, thrombocytopenia, elevated bilirubin and anorexia. CONCLUSION: Triple-drug chemotherapy achieves comparable results for response and survival to previously reported regimens, but with more toxicity. PMID- 19904268 TI - Chemoradiotherapy with concurrent gemcitabine and cisplatin with or without sequential chemotherapy with gemcitabine/cisplatin vs chemoradiotherapy with concurrent 5-fluorouracil in patients with locally advanced pancreatic cancer--a multi-centre randomised phase II study. AB - BACKGROUND: No standard treatment for locally advanced pancreatic cancer (LAPC) is defined. PATIENTS AND METHODS: Within a multi-centre, randomised phase II trial, 95 patients with LAPC were assigned to three different chemoradiotherapy (CRT) regimens: patients received conventionally fractionated radiotherapy of 50 Gy and were randomised to concurrent 5-fluorouracil (350 mg m(-2) per day on each day of radiotherapy, RT-5-FU arm), concurrent gemcitabine (300 mg m(-2)), and cisplatin (30 mg m(-2)) on days 1, 8, 22, and 29 (RT-GC arm), or the same concurrent treatment followed by sequential full-dose gemcitabine (1000 mg m(-2)) and cisplatin (50 mg m(-2)) every 2 weeks (RT-GC+GC arm). Primary end point was the overall survival (OS) rate after 9 months. RESULTS: The 9-month OS rate was 58% in the RT-5-FU arm, 52% in the RT-GC arm, and 45% in the RT-GC+GC arm. Corresponding median survival times were 9.6, 9.3, and 7.3 months (P=0.61) respectively. The intent-to-treat response rate was 19, 22, and 13% respectively. Median progression-free survival was estimated with 4.0, 5.6, and 6.0 months (P=0.21). Grade 3/4 haematological toxicities were more frequent in the two GC containing arms, no grade 3/4 febrile neutropaenia was observed. CONCLUSION: None of the three CRT regimens tested met the investigators' definition for efficacy; the median OS was similar to those previously reported with gemcitabine alone in LAPC. PMID- 19904269 TI - CITED2 and NCOR2 in anti-oestrogen resistance and progression of breast cancer. AB - BACKGROUND: Endocrine therapies of breast cancer are effective but ultimately fail because of the development of treatment resistance. We have previously revealed several genes leading to tamoxifen resistance in vitro by retroviral insertion mutagenesis. To understand the manner in which these genes yield tamoxifen resistance, their effects on global gene expression were studied and those genes resulting in a distinct gene expression profile were further investigated for their clinical relevance. METHODS: Gene expression profiles of 69 human breast cancer cell lines that were made tamoxifen resistant through retroviral insertion mutagenesis were obtained using oligonucleotide arrays and analysed with bioinformatic tools. mRNA levels of NCOR2 and CITED2 in oestrogen receptor-positive breast tumours were determined by quantitative RT-PCR. mRNA levels were evaluated for association with metastasis-free survival (MFS) in 620 patients with lymph node-negative primary breast cancer who did not receive systemic adjuvant therapy, and with clinical benefit in 296 patients receiving tamoxifen therapy for recurrent breast cancer. RESULTS: mRNA expression profiles of most tamoxifen-resistant cell lines were strikingly similar, except for the subgroups of cell lines in which NCOR2 or CITED2 were targeted by the retrovirus. Both NCOR2 and CITED2 mRNA levels were associated with MFS, that is, tumour aggressiveness, independently of traditional prognostic factors. In addition, high CITED2 mRNA levels were predictive for a clinical benefit from first-line tamoxifen treatment in patients with advanced disease. CONCLUSIONS: Most retrovirally targeted genes yielding tamoxifen resistance in our cell lines do not impose a distinctive expression profile, suggesting that their causative role in cell growth may be accomplished by post-transcriptional processes. The associations of NCOR2 and CITED2 with outcome in oestrogen receptor-positive breast cancer patients underscore the clinical relevance of functional genetic screens to better understand disease progression, which may ultimately lead to the development of improved treatment options. PMID- 19904271 TI - Age of onset in familial breast cancer as background data for medical surveillance. AB - BACKGROUND: Familial breast cancers are known to be of early onset. This article provides differences in the age of onset of breast cancer and death by breast cancer between women with and without a family history. METHODS: The Swedish Family-Cancer Database was used to estimate the cumulative risk of breast cancer and death by breast cancer according to family history with a stratified Cox model. Family history was defined separately for affected mother or sister considering their diagnostic ages. RESULTS: The age to reach the same cumulative incidence as women without family history decreased with decreasing diagnostic age of the affected relative. Women with a maternal history reached the risk of women lacking a family history at the age of 50 years between 12.3 (mother affected <40 years) and 3.3 years (mother affected >82 years) earlier. The trend for breast cancer mortality was essentially similar. CONCLUSIONS: Women with mother or sister affected by breast cancer are diagnosed and die at earlier ages than do women without family history. The differences depend on the diagnostic age of the affected relative. The present data may provide a rationale to derive recommendations for the starting age of screening in women with affected family members. PMID- 19904270 TI - A small molecule inhibitor of XIAP induces apoptosis and synergises with vinorelbine and cisplatin in NSCLC. AB - BACKGROUND: Evasion of apoptosis contributes to the pathogenesis of solid tumours including non-small cell lung cancer (NSCLC). Malignant cells resist apoptosis through over-expression of inhibitor of apoptosis proteins (IAPs), such as X linked IAP (XIAP). METHODS: A phenylurea-based small molecule inhibitor of XIAP, XIAP antagonist compound (XAC) 1396-11, was investigated preclincally to determine its ability to sensitise to clinically relevant cytotoxics, potentially allowing dose reduction while maintaining therapeutic efficacy. RESULTS: XIAP protein expression was detected in six NSCLC cell lines examined. The cytotoxicity of XAC 1396-11 against cultured NSCLC cell lines in vitro was concentration- and time-dependent in both short-term and clonogenic assays. XAC 1396-11-induced apoptosis was confirmed by PARP cleavage and characteristic nuclear morphology. XAC 1396-11 synergised with vinorelbine+/-cisplatin in H460 and A549 NSCLC cells. The mechanism of synergy was enhanced apoptosis, shown by increased cleavage of caspase-3 and PARP and by the reversal of synergy by a pan caspase inhibitor. Synergy between XAC 1396-11 and vinorelbine was augmented by optimising drug scheduling with superior effects when XAC 1396-11 was administered before vinorelbine. CONCLUSION: These preclinical data suggest that XIAP inhibition in combination with vinorelbine holds potential as a therapeutic strategy in NSCLC. PMID- 19904272 TI - Overdetection, overtreatment and costs in prostate-specific antigen screening for prostate cancer. AB - BACKGROUND: Prostate cancer screening with prostate-specific antigen (PSA) has shown to reduce prostate cancer mortality in the European Randomised study of Screening for Prostate Cancer (ERSPC) trial. Overdetection and overtreatment are substantial unfavourable side effects with consequent healthcare costs. In this study the effects of introducing widespread PSA screening is evaluated. METHODS: The MISCAN model was used to simulate prostate cancer growth and detection in a simulated cohort of 100,000 men (European standard population) over 25 years. PSA screening from age 55 to 70 or 75, with 1, 2 and 4-year-intervals is simulated. Number of diagnoses, PSA tests, biopsies, treatments, deaths and corresponding costs for 100,000 men and for United Kingdom and United States are compared. RESULTS: Without screening 2378 men per 100,000 were predicted to be diagnosed with prostate cancer compared with 4956 men after screening at 4-year intervals. By introducing screening, the costs would increase with 100% to 60,695,000 euro. Overdetection is related to 39% of total costs (23,669,000 euro). Screening until age 75 is relatively most expensive because of the costs of overtreatment. CONCLUSION: Introduction of PSA screening will increase total healthcare costs for prostate cancer substantially, of which the actual screening costs will be a small part. PMID- 19904273 TI - Preclinical mouse models for BRCA1-associated breast cancer. AB - A substantial part of all hereditary breast cancer cases is caused by BRCA1 germline mutations. In this review, we will discuss the insights into BRCA1 functions that we obtained from mouse models with conventional and conditional mutations in Brca1. The most advanced models closely resemble human BRCA1-related breast cancer and may therefore be useful for addressing clinically relevant questions. PMID- 19904274 TI - Abnormal expression of TRIB3 in colorectal cancer: a novel marker for prognosis. AB - BACKGROUND: TRIB3 is a human homologue of Drosophila tribbles. Previous studies have shown that TRIB3 controls the cell growth through ubiquitination-dependent degradation of other proteins, whereas its significance in the prognosis of colorectal cancer (CRC) is not yet fully understood. MATERIALS: This study comprised 202 patients who underwent surgery for CRC, as well as 22 cell lines derived from human gastrointestinal cancer. The correlation of gene expression with clinical parameters in patients was assessed. The biological significance was evaluated by knockdown experiments in seven colorectal cancer cell lines. RESULTS: A total of 20 cancer cell lines (90.9%) expressed the TRIB3 gene. The assessment in surgical specimens indicated that the gene expression was significantly higher in the cancerous region than in the marginal non-cancerous region. Patients with high TRIB3 expression were statistically susceptible to a recurrence of the disease, and showed poorer overall survival than those with low expression. The assessment of TRIB3 knockdown in five cell lines showed that small interfering RNA (siRNA) inhibition resulted in a statistically significant reduction in cell growth. CONCLUSION: These data strongly suggest the usefulness of TRIB3 as a marker for predicting the prognosis of CRC patients, showing a basis for the development of effective treatments for CRC. PMID- 19904275 TI - Humanised IgG1 antibody variants targeting membrane-bound carcinoembryonic antigen by antibody-dependent cellular cytotoxicity and phagocytosis. AB - BACKGROUND: The effect of glycoengineering a membrane specific anti carcinoembryonic antigen (CEA) (this paper uses the original term CEA for the formally designated CEACAM5) antibody (PR1A3) on its ability to enhance killing of colorectal cancer (CRC) cell lines by human immune effector cells was assessed. In vivo efficacy of the antibody was also tested. METHODS: The antibody was modified using EBNA cells cotransfected with beta-1,4-N acetylglucosaminyltransferase III and the humanised hPR1A3 antibody genes. RESULTS: The resulting alteration of the Fc segment glycosylation pattern enhances the antibody's binding affinity to the FcgammaRIIIa receptor on human immune effector cells but does not alter the antibody's binding capacity. Antibody-dependent cellular cytotoxicity (ADCC) is inhibited in the presence of anti-FcgammaRIII blocking antibodies. This glycovariant of hPR1A3 enhances ADCC 10-fold relative to the parent unmodified antibody using either unfractionated peripheral blood mononuclear or natural killer (NK) cells and CEA-positive CRC cells as targets. NK cells are far more potent in eliciting ADCC than either freshly isolated monocytes or granulocytes. Flow cytometry and automated fluorescent microscopy have been used to show that both versions of hPR1A3 can induce antibody-dependent cellular phagocytosis (ADCP) by monocyte-derived macrophages. However, the glycovariant antibody did not mediate enhanced ADCP. This may be explained by the relatively low expression of FcgammaRIIIa on cultured macrophages. In vivo studies show the efficacy of glycoengineered humanised IgG1 PR1A3 in significantly improving survival in a CRC metastatic murine model. CONCLUSION: The greatly enhanced in vitro ADCC activity of the glycoengineered version of hPR1A3 is likely to be clinically beneficial. PMID- 19904278 TI - A new perspective on the role of the orbitofrontal cortex in adaptive behaviour. AB - The orbitofrontal cortex (OFC) is crucial for changing established behaviour in the face of unexpected outcomes. This function has been attributed to the role of the OFC in response inhibition or to the idea that the OFC is a rapidly flexible associative-learning area. However, recent data contradict these accounts, and instead suggest that the OFC is crucial for signalling outcome expectancies. We suggest that this function--signalling of expected outcomes--can also explain the crucial role of the OFC in changing behaviour in the face of unexpected outcomes. PMID- 19904280 TI - Understanding microRNAs in neurodegeneration. AB - Interest in the functions of microRNAs (miRNAs) in the nervous system has recently expanded to include their roles in neurodegeneration. Investigations have begun to reveal the influence of miRNAs on both neuronal survival and the accumulation of toxic proteins that are associated with neurodegeneration, and are providing clues as to how these toxic proteins can influence miRNA expression. PMID- 19904279 TI - Plasticity and stability of visual field maps in adult primary visual cortex. AB - It is important to understand the balance between cortical plasticity and stability in various systems and across spatial scales in the adult brain. Here we review studies of adult plasticity in primary visual cortex (V1), which has a key role in distributing visual information. There are claims of plasticity at multiple spatial scales in adult V1, but a number of inconsistencies in the supporting data raise questions about the extent and nature of such plasticity. Our understanding of the extent of plasticity in V1 is further limited by a lack of quantitative models to guide the interpretation of the data. These problems limit efforts to translate research findings about adult cortical plasticity into significant clinical, educational and policy applications. PMID- 19904281 TI - Commentary on Perrone et al.: 'vitamin C: not for breakfast anymore...if you have myeloma'. PMID- 19904284 TI - Intrathecal corticoids in permanent focal cerebral ischemia in rats. Part I: a new therapeutic approach in the acute phase. AB - Intrathecally, triamcinolone acetonide (TCA) was suggested to have neuroprotective efficacy on infarction volume in acute focal cerebral ischemia in rats. In the first dose-finding study, TCA in five different doses or saline was administered into the cisterna magna of 12 rats, each 30 mins after endovascular occlusion of the middle cerebral artery (MCAO). In the second magnet resonance controlled confirmation study, the most neuroprotective dose was compared with controls in each of the 15 rats. Infarction volume was calculated at 24 h by 2.3.5 triphenyl-tetrazolium-chloride staining. Compared with controls (18.2%), infarction volume was significantly reduced using TCA at a dose of 0.012 mg/kg body weight (BW) (13.4%, P=0.04). TCA at doses of 0.03 (17.7%, P=0.84), 0.006 (15.9%, P=0.24), and 0.003 mg/kg BW (14.5%, P=0.11) did not significantly reduce infarction size. TCA 0.3 mg/kg BW resulted in bilateral infarction with increased infarction volume (19.8%, P=0.49). Magnetic resonance imaging confirmed successful MCAO and intrathecal administration. In experiment 2 compared with controls (20.0%), infarction volume was significantly reduced using TCA 0.012 mg/kg (13.4%, P=0.02). Intrathecally, TCA may significantly reduce infarction volume in acute focal cerebral ischemia in rats. Further studies are necessary to define the value of this therapy. PMID- 19904283 TI - Role of chemokines in CNS health and pathology: a focus on the CCL2/CCR2 and CXCL8/CXCR2 networks. AB - Chemokines and their receptors have crucial roles in the trafficking of leukocytes, and are of particular interest in the context of the unique immune responses elicited in the central nervous system (CNS). The chemokine system CC ligand 2 (CCL2) with its receptor CC receptor 2 (CCR2), as well as the receptor CXCR2 and its multiple ligands CXCL1, CXCL2 and CXCL8, have been implicated in a wide range of neuropathologies, including trauma, ischemic injury and multiple sclerosis. This review aims to overview the current understanding of chemokines as mediators of leukocyte migration into the CNS under neuroinflammatory conditions. We will specifically focus on the involvement of two chemokine networks, namely CCL2/CCR2 and CXCL8/CXCR2, in promoting macrophage and neutrophil infiltration, respectively, into the lesioned parenchyma after focal traumatic brain injury. The constitutive brain expression of these chemokines and their receptors, including their recently identified roles in the modulation of neuroprotection, neurogenesis, and neurotransmission, will be discussed. In conclusion, the value of evidence obtained from the use of Ccl2- and Cxcr2 deficient mice will be reported, in the context of potential therapeutics inhibiting chemokine activity which are currently in clinical trial for various inflammatory diseases. PMID- 19904285 TI - Measurement of density and affinity for dopamine D(2) receptors by a single positron emission tomography scan with multiple injections of [(11)C]raclopride. AB - Positron emission tomography (PET) with [(11)C]raclopride has been used to investigate the density (B(max)) and affinity (K(d)) of dopamine D(2) receptors related to several neurological and psychiatric disorders. However, in assessing the B(max) and K(d), multiple PET scans are necessary under variable specific activities of administered [(11)C]raclopride, resulting in a long study period and unexpected physiological variations. In this paper, we have developed a method of multiple-injection graphical analysis (MI-GA) that provides the B(max) and K(d) values from a single PET scan with three sequential injections of [(11)C]raclopride, and we validated the proposed method by performing numerous simulations and PET studies on monkeys. In the simulations, the three-injection protocol was designed according to prior knowledge of the receptor kinetics, and the errors of B(max) and K(d) estimated by MI-GA were analyzed. Simulations showed that our method could support the calculation of B(max) and K(d), despite a slight overestimation compared with the true magnitudes. In monkey studies, we could calculate the B(max) and K(d) of diseased or normal striatum in a 150 mins scan with the three-injection protocol of [(11)C]raclopride. Estimated B(max) and K(d) values of D(2) receptors in normal or partially dopamine-depleted striatum were comparable to the previously reported values. PMID- 19904286 TI - The adverse pial arteriolar and axonal consequences of traumatic brain injury complicated by hypoxia and their therapeutic modulation with hypothermia in rat. AB - This study examined the effect of posttraumatic hypoxia on cerebral vascular responsivity and axonal damage, while also exploring hypothermia's potential to attenuate these responses. Rats were subjected to impact acceleration injury (IAI) and equipped with cranial windows to assess vascular reactivity to topical acetylcholine, with postmortem analyses using antibodies to amyloid precursor protein to assess axonal damage. Animals were subjected to hypoxia alone, IAI and hypoxia, IAI and hypoxia before induction of moderate hypothermia (33 degrees C), IAI and hypoxia induced during hypothermic intervention, and IAI and hypoxia initiated after hypothermia. Hypoxia alone had no impact on vascular reactivity or axonal damage. Acceleration injury and posttraumatic hypoxia resulted in dramatic axonal damage and altered vascular reactivity. When IAI and hypoxia were followed by hypothermic intervention, no axonal or vascular protection ensued. However, when IAI was followed by hypoxia induced during hypothermia, axonal and vascular protection followed. When this same hypoxic insult followed the use of hypothermia, no benefit ensued. These studies show that early hypoxia and delayed hypoxia exert damaging axonal and vascular consequences. Although this damage is attenuated by hypothermia, this follows only when hypoxia occurs during hypothermia, with no benefit found if the hypoxic insult proceeds or follows hypothermia. PMID- 19904287 TI - Correlating tissue outcome with quantitative multiparametric MRI of acute cerebral ischemia in rats. AB - Predicting tissue outcome remains a challenge for stroke magnetic resonance imaging (MRI). In this study, we have acquired multiparametric MRI data sets (including absolute T(1), T(2), diffusion, T(1rho) using continuous wave and adiabatic pulse approaches, cerebral blood flow (CBF), and amide proton transfer ratio (APTR) images) during and after 65 mins of middle cerebral artery occlusion (MCAo) in rats. The MRI scans were repeated 24 h after MCAo, when the animals were killed for quantitative histology. Magnetic resonance imaging parameters acquired at three acute time points were correlated with regionally matching cell count at 24 h. The results emphasize differences in the temporal profile of individual MRI contrasts during MCAo and especially during early reperfusion, and suggest that complementary information from CBF and tissue damage can be obtained with appropriate MRI contrasts. The data show that by using three to four MRI parameters, sensitive to both hemodynamic changes and different aspects of parenchymal changes, the fate of the tissue can be predicted with increased correlation compared with single-parameter techniques. Combined multiparametric MRI data and multiparametric analysis may provide an excellent tool for preclinical testing of new treatments and also has the potential to facilitate decision-making in the management of acute stroke patients. PMID- 19904288 TI - Heat acclimation provides sustained improvement in functional recovery and attenuates apoptosis after traumatic brain injury. AB - Heat acclimation (HA) offers functional neuroprotection in mice after traumatic brain injury (TBI). This study further characterizes endogenous neuroprotection acquired by HA (34+/-1 degrees C, 30 d) after TBI. We establish here the ability of HA to induce sustained functional benefits and to reduce activation of apoptotic pathways. Neurobehavioral recovery, assessed by the Neurological Severity Score, was greater in HA mice up to 8 days after injury as compared with normothermic controls (P<0.05) and lesion volume was also smaller in the HA group (P<0.05). Reduced apoptotic cell death in HA mice was confirmed using caspase-3 activity measurements and immunohistochemistry. To investigate the underlying molecular pathways, expression levels of intrinsic apoptotic pathway-related proteins were examined. HA mice displayed higher mitochondrial levels of antiapoptotic Bcl-xL, accompanied by lower proapoptotic Bad levels and decreased cytochrome c release, suggesting a higher apoptotic threshold. Taken together with our previous reports, indicating increased Akt phosphorylation and antioxidative capacity, alongside with reduced tumor necrosis alpha levels after TBI in HA animals, the current results support the involvement of an antiapoptotic effect in HA-induced neuroprotection. Current results warrant further study as TBI-induced apoptosis may persist over weeks after injury, possibly providing a target for belated therapeutic intervention. PMID- 19904290 TI - Validity and reliability testing of a short questionnaire developed to assess consumers' use, understanding and perception of food labels. AB - BACKGROUND: Food labels may have an important function in communicating nutrition information and have considerable potential to influence food choice and dietary behaviour. OBJECTIVES: To assess the validity and reliability of a short (self complete) questionnaire designed to measure consumers' use and understanding of food labels. METHODS: Questionnaire content was determined by gaps highlighted in a literature review of food labelling. Nineteen questions (49 items) assessing frequency of label reading perceived importance of food labels, regularity of dining out, desire to have nutrition information at specific catering outlets and ability to perform nutrition information tasks were formulated and presented on four pages of A4. With the exception of two open-ended questions, all items were presented as closed (field box) structures. Content validity, face validity, item analysis, repeat and internal reliability were assessed. RESULTS: Nutrition experts (26) completed detailed content validity assessment, resulting in high scores for appropriateness, importance and phrasing of questions, although grammar and terminology changes were required. Face validity indicated that the questionnaire was quick to complete (<15 min), easy to follow and comprehensible. Cronbach's alpha scores (internal reliability) for questions with multiple sections ranged from 0.72 to 0.91, indicating good internal consistency. Repeat reliability testing showed Spearman's correlation coefficients ranging from 0.51 to 0.97 (all P<0.001) showing high temporal stability. Item Difficulty analysis indicated that questions (Section C only) were at an appropriate level (with P between 20 and 80% for all items). Item discrimination analysis ranged from r=0.43 to 0.70, highlighting that items were suitable for inclusion. CONCLUSIONS: This questionnaire is a suitable tool for assessing consumers' use, understanding and perception of food labels. PMID- 19904289 TI - Antioxidant CR-6 protects against reperfusion injury after a transient episode of focal brain ischemia in rats. AB - Oxidative and nitrosative stress are targets for intervention after ischemia/reperfusion. The aim of this study was to explore the effect of CR-6, a vitamin-E analogue that is antioxidant and scavenger of nitrogen-reactive species. Sprague-Dawley rats had the middle cerebral artery (MCA) occluded either for 90 mins or permanently. Cortical perfusion was continuously monitored by laser-Doppler flowmetry. CR-6 (100 mg/kg) was administered orally either at 2 and 8 h after MCA occlusion, or at 2 h only. Infarct volume, neurological deficit, and signs of reperfusion injury were evaluated. CR-6 was detected in plasma and brain by HPLC. CR-6 reduced glutathione consumption in the ischemic brain and superoxide generation in the isolated MCA. CR-6 decreased infarct volume and attenuated the neurological deficit at 1 and 7 days after ischemia/reperfusion, but not after permanent ischemia. Immediately after reperfusion, cortical blood flow values returned to their baseline (+/-20%) in several animals, whereas others showed hyper-perfusion (>20% of baseline). Reactive hyperemia was associated with adverse events such as increased cortical BBB leakage, edema, protein nitrotyrosination, COX-2 expression, and neutrophil accumulation; and with a poorer outcome, and CR-6 attenuated these effects. In conclusion, oral CR 6 administration after transient ischemia protects the brain from reperfusion injury. PMID- 19904291 TI - Relationship between body mass index and alanine aminotransferase concentration in non-diabetic Korean adults. AB - BACKGROUND/OBJECTIVES: Obesity has been identified as an important factor of elevated alanine aminotransferase (ALT), a surrogate of non-alcoholic fatty liver disease. We investigated the relationship between obesity and elevated ALT in the general Korean population. SUBJECTS/METHODS: The study sample was comprised of 3098 Korean adults who had participated in the 2005 Korean National Health and Nutrition Examination Survey. Obesity was assessed using body mass index (BMI). Overweight (23< or =BMI<25) and obese (BMI> or =25) were defined by the International Obesity Task Force for Asian adults. ALT was measured by enzymatic methods. Elevated ALT was defined as an enzyme activity >40 IU/l for men and >31 IU/l for women. RESULTS: Among participants, 246 (12.7%) men and 86 (7.4%) women displayed elevated ALT. BMI was significantly higher in men and women with elevated ALT than those with normal ALT (P<0.0001). The unadjusted odds ratio (OR) for elevated ALT increased according to the degree of BMI in men (OR: 1.9, 95% confidence interval (CI): 1.25-2.93 in overweight vs OR: 5.01, 95% CI: 3.49 7.21 in obese; P<0.0001) and women (OR: 2.44, 95% CI: 1.24-4.82 vs OR: 3.94, 95% CI: 2.18-7.13; P<0.0001). This trend did not differ after adjustment for putative risk factors including alcohol intake in men (OR: 1.56 vs OR: 3.47, P<0.0001) and women (OR: 1.55 vs OR: 3.10, P=0.0015). CONCLUSION: BMI is implicated as a strong risk factor of elevated ALT in non-diabetic Korean adults. PMID- 19904292 TI - Dietary antioxidant capacity and concentration of adiponectin in apparently healthy adults: the ATTICA study. AB - BACKGROUND/OBJECTIVES: This study aimed at evaluating the relationship of adiponectin concentration with total dietary antioxidant capacity in free-living, apparently healthy adults from the ATTICA study. SUBJECTS/METHODS: A random subsample from the ATTICA study, consisting of 310 men (40+/-11 years) and 222 women (38+/-12 years), was selected. Adiponectin, along with other inflammatory markers, was measured in fasting participants. Dietary habits were evaluated using a food frequency questionnaire and the dietary antioxidant capacity was based on published values of Italian foods measured by three different assays: ferric-reducing antioxidant power (FRAP), total radical-trapping antioxidant parameters (TRAP) and trolox-equivalent antioxidant capacity (TEAC). RESULTS: Positive associations were observed between dietary antioxidant capacity and adiponectin concentration, as assessed with FRAP (b+/-s.e.=0.012+/-0.005, P=0.018 per 1 mmol Fe (II)/day), TRAP (b+/-s.e.=0.030+/-0.013, P=0.017 per 1 mmol trolox equivalent/day) and TEAC (b+/-s.e.=0.025+/-0.012, P=0.042, per 1 mmol trolox equivalent/day) in multiadjusted analysis. Moreover, a negative relation of dietary antioxidant indices with inflammatory markers was revealed. CONCLUSIONS: Diets with high antioxidant capacity are related to increased adiponectin levels. An adiponectin-mediated route through which antioxidant-rich foods exert beneficial effects against inflammation and cardiovascular diseases can be thus hypothesized. PMID- 19904293 TI - Supplementing iron and zinc: double blind, randomized evaluation of separate or combined delivery. AB - BACKGROUND/OBJECTIVES: Many children have diets deficient in both iron and zinc, but there has been some evidence of negative interactions when they are supplemented together. The optimal delivery approach would maximize clinical benefits of both nutrients. We studied the effectiveness of different iron and zinc supplement delivery approaches to improve diarrhea and anemia in a rural Bangladesh population. STUDY DESIGN: Randomized, double blind, placebo-controlled factorial community trial. RESULTS: Iron supplementation alone increased diarrhea, but adding zinc, separately or together, attenuated these harmful effects. Combined zinc and iron was as effective as iron alone for iron outcomes. All supplements were vomited <1% of the time, but combined iron and zinc were vomited significantly more than any of the other supplements. Children receiving zinc and iron (together or separately) had fewer hospitalizations. Separating delivery of iron and zinc may have some additional benefit in stunted children. CONCLUSIONS: Separate and combined administration of iron and zinc are equally effective for reducing diarrhea, hospitalizations and improving iron outcomes. There may be some benefit in separate administration in stunted children. PMID- 19904294 TI - A technique to assess body composition and sarcopenia using DXA: application for an obese population. AB - The objective of this study was to explore the accuracy of summing multiple dual energy X-ray absorptiometry (DXA) scans to estimate whole-body soft tissue measures and the sarcopenic index in female subjects for application to an obese population. Forty-five women who fit within the DXA scanning region (M+/-s.d.; BMI=24.5+/-4.6 kg/m(2)) were scanned four times (one normal whole body (WB) scan, one head/trunk/leg scan allowing trunk delineation (TRK-H-L), one scan with body shifted right (LA) and one left (RA) to allow arm delineation). Fat mass (FM) and appendicular lean mass were determined from the WB scan (aLM(WB)), with the latter determined by summing arm and leg mineral-free lean mass (MFLM). Strong agreement and no differences (M+/-s.d.(DIFF)) were found between FM(WB) and FM(SUM) (-0.12+/-0.38 kg, P=0.37; r=0.999, P<0.001); MFLM(WB) and MFLM(SUM) (0.02+/-0.36 kg, P=0.80; r=0.998, P<0.001); and aLM(WB) and aLM(SUM) (0.12+/-0.32 kg, P=0.63; r=0.994, P<0.001). Summing DXA scans is a valid method for determining the risk for sarcopenic obesity and may aid research regarding obesity and risk for disability. PMID- 19904295 TI - BMI at 4 years of age is associated with previous and current protein intake and with paternal BMI. AB - OBJECTIVES: To evaluate possible associations between body mass index (BMI) at 4 years of age, current and previous dietary intakes and parental BMI. METHODS: A follow-up of dietary intake and anthropometry in 127 4-year-old children corresponding to 54% of children who completed an initial intervention study at 18 months of age. RESULTS: Fourteen percent of the girls and 13% of the boys were overweight (age-adjusted BMI> or =25) and 2% of the girls and 3% of the boys were obese (age-adjusted BMI> or =30). Thirty-four percent and 9% of the fathers and 19 and 7% of the mothers were overweight and obese, respectively. BMI at 6-18 months was a strong predictor of BMI at 4 years. Univariate regression analyses revealed that intake of protein in particular, and also of total energy and carbohydrates at 17/18 months and at 4 years, was positively associated with BMI at 4 years. Although BMI at 6-18 months was the strongest predictor of BMI at 4 years, in the final multivariate models of the child's BMI, protein intake at 17 18 months and at 4 years, energy intake at 4 years and the father's-but not the mother's-BMI were also independent contributing factors. CONCLUSIONS: Among these healthy children, BMI at 4 years of age tracked from 6 to 18 months of age and were associated with previous and current protein intake as well as parental BMI, particularly that of the father. PMID- 19904296 TI - Consumption of animal products, their nutrient components and postmenopausal circulating steroid hormone concentrations. AB - BACKGROUND/OBJECTIVES: Little is known about nutritional factors that influence circulating concentrations of steroid hormones, which are consistently associated with risk of breast cancer for postmenopausal women. We aimed to investigate the association between consumption of animal products and the plasma concentrations of steroid hormones and sex hormone-binding globulin (SHBG). SUBJECTS/METHODS: Cross-sectional analysis was conducted on plasma from 766 naturally postmenopausal women. We measured plasma concentrations of steroid hormones and SHBG, and estimated dietary intakes using a 121-item food frequency questionnaire. Log-transformed values of hormone concentrations were regressed on quartiles of intake of meat and dairy products among food items, and fats, proteins and cholesterol among nutrient intake. RESULTS: Total red and fresh red meat consumption was negatively associated with SHBG levels (P for trend=0.04 and <0.01, respectively). Mean SHBG concentrations were approximately 8% and 13% lower for women in the highest quartile compared with the lowest quartile of total red and fresh red meat consumption, respectively. Positive associations were observed between dairy product consumption and total and free estradiol concentrations (P for trend=0.02 and 0.03, respectively). Mean concentrations of total and free estradiol were 15 and 14% higher for women in the highest quartile of dairy product consumption than for those in the lowest quartile, respectively. No associations were observed with consumption of processed meat, chicken, fish, eggs, cholesterol, fats or protein. CONCLUSIONS: Our study suggests that greater consumption of total red and fresh red meat and dairy products might influence circulating concentrations of SHBG and estradiol, respectively. Confirmation and further investigation is required. PMID- 19904297 TI - Experimental designs for evaluation of genetic variability and selection of ancient grapevine varieties: a simulation study. AB - Classical methodologies for grapevine selection used in the vine-growing world are generally based on comparisons among a small number of clones. This does not take advantage of the entire genetic variability within ancient varieties, and therefore limits selection challenges. Using the general principles of plant breeding and of quantitative genetics, we propose new breeding strategies, focussed on conservation and quantification of genetic variability by performing a cycle of mass genotypic selection prior to clonal selection. To exploit a sufficiently large amount of genetic variability, initial selection trials must be generally very large. The use of experimental designs adequate for those field trials has been intensively recommended for numerous species. However, their use in initial trials of grapevines has not been studied. With the aim of identifying the most suitable experimental designs for quantification of genetic variability and selection of ancient varieties, a study was carried out to assess through simulation the comparative efficiency of various experimental designs (randomized complete block design, alpha design and row-column (RC) design). The results indicated a greater efficiency for alpha and RC designs, enabling more precise estimates of genotypic variance, greater precision in the prediction of genetic gain and consequently greater efficiency in genotypic mass selection. PMID- 19904298 TI - Integrins: masters and slaves of endocytic transport. AB - Since it has become clear that adhesion receptors are trafficked through the endosomal pathway and that this can influence their function, much effort has been invested in obtaining detailed descriptions of the molecular machinery responsible for internalizing and recycling integrins. New findings indicate that integrin trafficking dictates the nature of Rho GTPase signalling during cytokinesis and cell migration. Furthermore, integrins can exert control over the trafficking of other receptors in a way that drives cancer cell invasion and tumour angiogenesis. PMID- 19904299 TI - Single-sperm analysis for recurrence risk assessment of spinal muscular atrophy. AB - With the detection of a homozygous deletion of the survival motor neuron 1 gene (SMN1), prenatal and preimplantation genetic diagnosis (PGD) for spinal muscular atrophy has become feasible and widely applied. The finding of a de novo rearrangement, resulting in the loss of the SMN1 gene, reduces the recurrence risk from 25% to a lower percentage, the residual risk arising from recurrent de novo mutation or germline mosaicism. In a couple referred to our PGD center because their first child was affected with SMA, the male partner was shown to carry two SMN1 copies. An analysis of the SMN1 gene and two flanking markers was performed on 12 single spermatozoa, to determine whether the father carried a CIS duplication of the SMN1 gene on one chromosome and was a carrier, or if the deletion has occurred de novo. We showed that all spermatozoa that were carriers of the 'at-risk haplotype' were deleted for the SMN1 gene, confirming the carrier status of the father. We provide an original application of single germ cell studies to recessive disorders using coamplification of the gene and its linked markers. This efficient and easy procedure might be useful to elucidate complex genetic situations when samples from other family members are not available. PMID- 19904300 TI - Hereditary haemorrhagic telangiectasia and genetic thrombophilia. PMID- 19904302 TI - Haploinsufficiency of MBD5 associated with a syndrome involving microcephaly, intellectual disabilities, severe speech impairment, and seizures. AB - Microdeletion of chromosome 2q23.1 results in a novel syndrome previously reported in five individuals. Many of the del(2)(q23.1) cases were thought to have other syndromes such as Angelman, Prader-Willi, or Smith-Magenis because of certain overlapping clinical features. We report two new cases of the 2q23.1 microdeletion syndrome, describe the syndrome phenotype, define the minimal critical region, and analyze the expression of critical region genes toward identification of the causative gene(s) for the disorder. Individuals with del(2)(q23.1) have severe developmental and cognitive delays, minimal speech, seizures, microcephaly, mild craniofacial dysmorphism, behavioral disorders, and short stature. The deletions encompassing 2q23.1 range from >4 Mb to <200 kb, as identified by oligonucleotide and BAC whole-genome array comparative hybridization. The minimal critical region includes a single gene, MBD5, deleted in all cases, whereas all but one case also include deletion of EPC2. Quantitative real-time PCR of patient lymphoblasts/lymphocytes showed an approximately 50% reduced expression of MBD5 and EPC2 compared with controls. With similar phenotypes among the 2q23.1 deletion patients, the idea of one or more common genes causing the pathological defect seen in these patients becomes evident. As all five previous cases and the two cases in this report share one common gene, MBD5, we strongly suspect that haploinsufficiency of MBD5 causes most of the features observed in this syndrome. PMID- 19904303 TI - Sensitivity enhancement of fiber loop cavity ring-down pressure sensor. AB - We present a theoretical and experimental study on sensitivity enhancement of a fiber-loop cavity ring-down pressure sensor. The cladding of the sensing fiber is etched in hydrofluoric acid solution to enhance its sensitivity. The experimental results demonstrate that the pressure applied on the sensing fiber is linearly proportional to the difference between the reciprocals of the ring-down time with and without pressure, and the relative sensitivity exponentially increases with decreasing the cladding diameter. When the sensing fiber is etched to 41.15 microm, its sensitivity is about 36 times that of nonetched fiber in the range of 0 to 32.5 MPa. The measured relative standard deviation of the ring-down time is about 0.15% and, correspondingly, the least detectable loss is about 0.00069 dB. PMID- 19904304 TI - Adaptive thresholding and dynamic windowing method for automatic centroid detection of digital Shack-Hartmann wavefront sensor. AB - A Shack-Hartmann wavefront sensor (SWHS) splits the incident wavefront into many subsections and transfers the distorted wavefront detection into the centroid measurement. The accuracy of the centroid measurement determines the accuracy of the SWHS. Many methods have been presented to improve the accuracy of the wavefront centroid measurement. However, most of these methods are discussed from the point of view of optics, based on the assumption that the spot intensity of the SHWS has a Gaussian distribution, which is not applicable to the digital SHWS. In this paper, we present a centroid measurement algorithm based on the adaptive thresholding and dynamic windowing method by utilizing image processing techniques for practical application of the digital SHWS in surface profile measurement. The method can detect the centroid of each focal spot precisely and robustly by eliminating the influence of various noises, such as diffraction of the digital SHWS, unevenness and instability of the light source, as well as deviation between the centroid of the focal spot and the center of the detection area. The experimental results demonstrate that the algorithm has better precision, repeatability, and stability compared with other commonly used centroid methods, such as the statistical averaging, thresholding, and windowing algorithms. PMID- 19904305 TI - Polarization-gated imaging in tissue phantoms: effect of size distribution. AB - We have investigated the effect of size distribution of aqueous solutions of monodisperse and a mixture of polydisperse scatterers of two different sizes on the image quality using linear and circularly polarized light. The contrast and resolution are affected by the size distribution present in the mixture of a polydisperse medium, while they are affected by the refractive index in a monodisperse medium. Circularly polarized light improves image quality of polydisperse scatterers. Images in the polydisperse medium are retrieved for values of optical thickness less than those of the large-sized monodisperse medium. We offer plausible explanations for all the experimental observations. PMID- 19904306 TI - Long-term stabilization of a heterodyne metrology interferometer down to a noise level of 20 pm over an hour. AB - A heterodyne metrology interferometer was stabilized down to a noise level of 20 picometers (pm) as a root-mean-square (RMS) value integrated between 0.3 mHz and 1 Hz. This noise level was achieved by employing active and passive interferometer stabilization techniques. The heterodyne interferometer was built on a 50 mm square ultralow expansion glass plate in order to reduce an optical path length change caused by temperature variation. An optical configuration of the interferometer is a Mach-Zehnder interferometer with a design as symmetric as possible so that a detection signal can be insensitive to homogeneous thermal expansion of the glass plate. The heterodyne frequency is actively controlled in order to suppress residual noises caused by optical path length changes outside of the glass plate as well as phase fluctuations of the heterodyne frequency source. Our stabilization scheme is considered useful in achieving the 20 pm noise level without a stable heterodyne frequency source, as well as temperature stabilization around a whole apparatus. This interferometer can be used in precise metrology applications, such as characterization of deformation for satellite optical components against thermal exposure. PMID- 19904307 TI - Bicoherence: a new lucky region technique in anisoplanatic image restoration. AB - A novel algorithm is proposed to predict "lucky" regions in a sequence of long range camera images affected by atmospheric turbulence. Our new approach is to employ bicoherence as a measure of quality to determine lucky regions or good quality image patches from a recorded sequence of anisoplanatic images. The better-quality image regions are selected according to the magnitude of the average value of the bicoherence of each region. Each image patch is restored using bispectral phase estimation from lucky regions, before mosaicing to an overall restoration. Bicoherence can also be used to predict lucky images in the isoplanatic case. Experiments show that our algorithm performs well with both simulated and naturally degraded data. PMID- 19904308 TI - Around-the-objective total internal reflection fluorescence microscopy. AB - Total internal reflection fluorescence (TIRF) microscopy uses the evanescent field on the aqueous side of a glass/aqueous interface to selectively illuminate fluorophores within approximately 100 nm of the interface. Applications of the method include epi-illumination TIRF, where the exciting light is refracted by the microscope objective to impinge on the interface at incidence angles beyond critical angle, and prism-based TIRF, where exciting light propagates to the interface externally to the microscope optics. The former has higher background autofluorescence from the glass elements of the objective where the exciting beam is focused, and the latter does not collect near-field emission from the fluorescent sample. Around-the-objective TIRF, developed here, creates the evanescent field by conditioning the exciting laser beam to propagate through the submillimeter gap created by the oil immersion high numerical aperture objective and the glass coverslip. The approach eliminates background light due to the admission of the laser excitation to the microscopic optics while collecting near field emission from the dipoles excited by the approximately 50 nm deep evanescent field. PMID- 19904309 TI - Digital simulation of scalar optical diffraction: revisiting chirp function sampling criteria and consequences. AB - Accurate simulation of scalar optical diffraction requires consideration of the sampling requirement for the phase chirp function that appears in the Fresnel diffraction expression. We describe three sampling regimes for FFT-based propagation approaches: ideally sampled, oversampled, and undersampled. Ideal sampling, where the chirp and its FFT both have values that match analytic chirp expressions, usually provides the most accurate results but can be difficult to realize in practical simulations. Under- or oversampling leads to a reduction in the available source plane support size, the available source bandwidth, or the available observation support size, depending on the approach and simulation scenario. We discuss three Fresnel propagation approaches: the impulse response/transfer function (angular spectrum) method, the single FFT (direct) method, and the two-step method. With illustrations and simulation examples we show the form of the sampled chirp functions and their discrete transforms, common relationships between the three methods under ideal sampling conditions, and define conditions and consequences to be considered when using nonideal sampling. The analysis is extended to describe the sampling limitations for the more exact Rayleigh-Sommerfeld diffraction solution. PMID- 19904310 TI - Measurement of axial and transverse trapping stiffness of optical tweezers in air using a radially polarized beam. AB - The trapping efficiency and stiffness of optical tweezers using radial polarization are evaluated; the ray-tracing method and a proposed measurement method are used for numerical and experimental analyses, respectively. The maximum axial trapping efficiency with radial polarization is 1.84 times that with linear polarization, while the maximum transverse trapping efficiency decreases by 0.58 times. Further, the axial and transverse trapping efficiencies are found to be 1.19 times larger and 0.83 times smaller, respectively, than the values with linear polarization. From the experiments, the axial and transverse stiffness values are 1.2 times larger and 0.8 times smaller, respectively, with radial polarization. Hence, radial polarization enhances the axial trapping properties while reducing the transverse trapping properties. PMID- 19904311 TI - Imaging capability of patterned liquid crystals. AB - We demonstrate the ability to make high resolution arbitrary patterned optical retarders using liquid crystal polymer (LCP). Contact lithography is used to define unique LCP alignment domains. Patterned LCP retarders are imaged between crossed polarizers to determine pattern visibility as a function of feature size. It was determined that patterned retarders for wavelengths between 250 nm and 2500 nm can be constructed with feature sizes as small as 4 microm. We also showed that multiple patterns can be created on the same substrate using a combination of patterned LCP and opaque features. Our process has applications in displays, double-patterning lithography, and imaging polarimetry. PMID- 19904312 TI - Monte Carlo simulation of multiple photon scattering in sugar maple tree canopies. AB - Detecting objects hidden beneath forest canopies is a difficult task for optical remote sensing systems. Rather than relying upon the existence of gaps between leaves, as other researchers have done, our ultimate goal is to use light scattered by leaves to image through dense foliage. Herein we describe the development of a Monte Carlo model for simulating the scattering of light as it propagates through the leaves of an extended tree canopy. We measured several parameters, including the gap fraction and maximum leaf-area density, of a nearby sugar maple tree grove and applied them to our model. We report the results of our simulation in both the ground and the receiver planes for an assumed illumination angle of 80 degrees. To validate our model, we then illuminated the sugar maple tree grove at 80 degrees and collected data both on the canopy floor and at our monostatic receiver aperture. Experimental results were found to correlate well with our simulated expectations. PMID- 19904313 TI - Microgrinding of lensed fibers by means of a scanning-probe microscope setup. AB - We describe a precision grinding procedure that allows treating a previously etched fiber tip to conform to predefined shapes, including hemispherical and axial conical (axicon) lenses. The grinding method is based on mechanical polishing with the fiber tip moving in a translational mode inside a conical polishing surface. The grinding procedure is performed in a homemade scanning probe microscope equipped with a shear-force sensor based on a piezoelectric tuning fork as well as with capacitor position sensors. The scanning probe microscope is operated either as atomic force microscope for topographic characterization of the tip shape and the polishing surface or as a scanning near field microscope for measurement of the light focusing properties of the ground microlenses. PMID- 19904314 TI - Retrieval of size and refractive index of spherical particles by multiangle light scattering: neural network method application. AB - A method to retrieve the radius and the relative refractive index of spherical homogeneous nonabsorbing particles by multiangle scattering is proposed. It is based on the formation of noise-resistant functionals of the scattered intensity, which are invariant with respect to the linear homogeneous transformations of an intensity-based signal and approximation of the retrieved parameters' dependence on the functionals by a feed-forward neural network. The neural network was trained by minimization of the mean squared relative error in the range of particle radii from 0.6 mkm up to 13.6 mkm and relative refractive index from 1.015 up to 1.28. In comparison with training on a minimum of the mean squared error, this method enables one to increase the accuracy of the radius retrieval in the range of radii from 0.6 to 2 microm and refractive index in the range from 1.015 to 1.1. The values of intensity of light scattered in the interval of angles 10 degrees-60 degrees are used as input data. If the measurement error is 20%, the mean errors of the radius and relative refractive index are 0.8% and 7%, respectively. The results obtained by the proposed method and by the trial and error method with published experimental data (measured with a scanning flow cytometer) are compared. The maximal difference in the retrieval results of radius and the relative refractive index of particles obtained by both methods is under 5%. PMID- 19904315 TI - Diode-laser-based high-precision absolute distance interferometer of 20 m range. AB - We present a hybrid absolute distance measurement method that is based on a combination of frequency sweeping, variable synthetic, and two-wavelength, fixed synthetic wavelength interferometry. Both experiments were realized by two external cavity diode lasers. The measurement uncertainty was experimentally and theoretically demonstrated to be smaller than 12 microm at a measurement distance of 20 m. PMID- 19904316 TI - Improved differential confocal microscopy with ultrahigh signal-to-noise ratio and reflectance disturbance resistibility. AB - Improved differential confocal microscopy is proposed to improve axial resolution and to enhance disturbance resistibility of confocal microscopy. The subtraction and sum values of the two defocusing detected signals are divided as the response function. Both ultrahigh signal-to-noise ratio (SNR) and wide range can be selectively obtained by controlling the defocusing amount of the two differential detectors more tightly with the reflectance disturbance resistibility. Since the detecting sensitivity of the proposed confocal microscopy is unrelated to the energy loss of the reflected beam, the multiplicative mode disturbance can be used to measure microstructures made of hybrid materials and overcome the power drift of a laser source during long scanning. In the case of ultrahigh SNR, the axial resolution reaches 1 nm when NA=0.75 and lambda=632.8 nm. PMID- 19904317 TI - Beam quality degradation of a higher order transverse mode beam due to spherical aberration of a lens. AB - An analysis of the increase in the beam-propagation factor, M2, of a higher order transverse mode beam caused by quartic phase aberration after transmission through a spherically aberrated lens is reported. The analysis shows that for a given beam size, the increase in the M2 parameter is less for a higher order transverse mode beam as compared to that for a diffraction limited beam. Experimental results using a multimode laser diode beam show good agreement with the theoretical results. PMID- 19904318 TI - Fabrication and transmission characteristics of infrared hollow fiber based on silver-clad stainless steel pipes. AB - Silver-clad stainless steel pipe is used as the supporting tube for the fabrication of infrared hollow fiber. The hollow fiber has high mechanical strength and is highly durable for use in the medical sterilization process. Film of a cyclic olefin polymer layer or silver iodide (AgI) was coated internally to reduce the transmission loss. A liquid-filling method is proposed for coating the AgI layer. Multiple coating processes proved to be effective to increase the AgI film thickness. A treatment of sodium thiosulfate water solution is also proposed to reduce the film thickness. The film thickness can be accurately controlled by combining the coating and decoating techniques. A loss of less than 0.2 dB was obtained for CO2 laser light for a hollow pipe with a length of 280 mm and an inside diameter of 0.75 mm. PMID- 19904319 TI - Design of flat-top interleaver and tunable dispersion compensator using cascaded Sagnac loop mirrors and ring resonators. AB - We propose multicavity reflective Gires-Tournois etalons (MCR-GTE) and generalized multicavity transmissive Gires-Tournois etalons (GMCT-GTE) composed of cascaded Sagnac loop mirrors and ring resonators, respectively. Michelson Gires-Tournois interferometers (MGTI) based on two sets of MCR-GTEs are theoretically studied. As the focal point, we demonstrate the GMCT-GTE, which is a reciprocal and transmissive element as a tunable dispersion compensator (TDC) for the proposed interleaver. The reciprocal and transmissive TDC is superior to reflective ones in terms of saving the number of TDCs. Only one set of TDCs is sufficient for the two output ports of the MGTI proposed. Discussions on fabrication tolerances are given as well, focusing on two main factors that degrade the performance of the interleaver and its TDC in practice, i.e., the length mismatch and loss. PMID- 19904320 TI - Broadband optical pulse scattering from a glass fiber. AB - We report results of scattering measurements using femtosecond pulses to collect a wealth of information in a single experiment. Potential issues with particle scattering, such as variation in particle size, were avoided by using 9 and 50 microm diameter glass fibers. We first establish an angular scattering intensity baseline, and we show that the spectral width of very short pulses leads to smoothing of the angular scattering pattern, consistent with continuous broadband illumination. We then measure the angular scattering pattern from short pulses with a spectrometer and reveal an underlying spectral periodicity of broadband scattered light that is consistent with narrowband cw scattering. Our experimental results compare well with existing theory. We show that such two dimensional experimental data and derived analytic solution can provide robust characterization of scattering objects even in the presence of noise. PMID- 19904321 TI - Theoretical analysis of the image with a local intensity minimum during hot image formation in high-power laser systems. AB - The evolution of an image with a local intensity minimum (LIM image) during hot image formation caused by translucent obscuration is theoretically and numerically investigated for the first time to our knowledge. The existence of the LIM image is proved, and the functional relationship describing the intensity of the LIM image is derived by using transfer matrix theory. Furthermore, the influences of the parameters of the obscuration and the nonlinear medium on the intensity of the LIM image are also discussed. The results show that the intensity of the LIM image increases with the increase of the amplitude coefficients of the obscuration, but declines with increasing the thickness of the nonlinear medium within a certain scope. We also found that the distance from the rear surface of the nonlinear medium to the LIM image plane is approximately equal to that from the obscuration plane to the rear surface of the nonlinear medium. PMID- 19904322 TI - Improving signal-to-noise ratio by use of a cross-shaped aperture in the holographic data storage system. AB - A cross-shaped aperture is proposed to improve signal-to-noise ratio (SNR) in the holographic data storage system (HDSS). Both simulated and experimental results show that higher SNR can be achieved by the cross-shaped aperture than traditional square or circular apertures with the same area. A maximum gain of 20% in SNR is obtained for the optimized cross-shaped aperture. The sensitivities to pixel misalignment and magnification error are also numerically compared. PMID- 19904324 TI - Self-images location of amplitude/phase binary gratings. AB - We analyze the near-field behavior of binary amplitude/phase diffraction gratings, which modulate at the same time the amplitude and phase of the incident light beam. As it is expected, the distance between two consecutive self-images of the grating depends only on the period of the grating and the wavelength of the illumination. However, the location of the self-images depends on the specific properties of the grating. In this work, we analyze the location of the self-images in terms of the Fourier coefficients of the grating, obtaining analytical expressions. This analysis can be useful in applications in which the position of the self-images must be at certain fixed distances from the grating. Finally, an experimental and numerical verification of the proposed theory is performed. PMID- 19904323 TI - Long-range time-of-flight scanning sensor based on high-speed time-correlated single-photon counting. AB - We describe a scanning time-of-flight system which uses the time-correlated single-photon counting technique to produce three-dimensional depth images of distant, noncooperative surfaces when these targets are illuminated by a kHz to MHz repetition rate pulsed laser source. The data for the scene are acquired using a scanning optical system and an individual single-photon detector. Depth images have been successfully acquired with centimeter xyz resolution, in daylight conditions, for low-signature targets in field trials at distances of up to 325 m using an output illumination with an average optical power of less than 50 microW. PMID- 19904325 TI - Design considerations for low-light level low-Fresnel number optical systems. AB - Low-Fresnel number optical systems exhibit significant diffraction effects that cause a shift in the peaks of on-axis irradiance away from the geometric focal point. This is currently interpreted as a change of the focal length of an optical system, leading optical system designers to compensate for the effect by assuming the image plane is coincident with the peak of on-axis irradiance. While this may be an appropriate interpretation for certain applications, I show that despite the shift in peak irradiance away from the geometrical focal point, a change in a system's optical power will not increase the on-axis irradiance at that distance. This is important for low-light level applications where it is necessary to mitigate diffraction induced transmission losses. I also show that low-Fresnel number systems have increased tolerance on system power at the geometrical focal point and as a result are inherently achromatic. PMID- 19904326 TI - Optical current sensor immune to reflection phase shift based on graded-index magneto-optical glass. AB - We propose a new design of bulk glass optical current sensor immune to reflection phase shift, in which an annular graded-index magneto-optical glass with a small prism is used as a sensing head, and the inner and outer layers of the glass possess gradient refractive indices, while the center layer is uniform. Our theoretical analyses show that under certain conditions the light beam will no longer reach the interface between the magneto-optical glass and air during the propagation in the sensing head. Therefore the reflection phase shift could be avoided essentially, resulting in dramatic enhancement of the sensitivity. The influences of the geometrical parameters on the beam traces and the effective range of the initial angle are specified by numerical simulations. PMID- 19904327 TI - Propagation-based phase-contrast enhancement of nanostructure images using a debris-free femtosecond-laser-driven cluster-based plasma soft x-ray source and an LiF crystal detector. AB - We demonstrate in-line phase-contrast imaging of nanothickness foils by using a relatively large, polychromatic, debris-free femtosecond-laser-driven cluster based plasma soft x-ray source, and a high-resolution, large dynamic range LiF crystal detector. The spatial coherence length of radiation in our setup reached a value of 5 microm on the sample plane, which is enough to observe phase contrast enhancement in the images registered by the detector placed only a few hundred micrometers behind the object. We have developed a tabletop soft x-ray emission source, which emits radiation within a 4pi sr solid angle, and which allows one to obtain contact and propagation-based phase-contrast imaging of nanostructures with 700 nm spatial resolutions. This advance could be of utility for metrology applications. PMID- 19904328 TI - Automatic null ellipsometry with an interferometer. AB - A new approach to automatic null ellipsometry is described in which the analyzer of a traditional polarizer compensator sample analyzer (PCSA) null ellipsometer is replaced with a heterodyne Michelson interferometer. One arm of this interferometer is modified such that it produces a fixed, linearly polarized reference beam, irrespective of the input polarization state. This beam is recombined interferometrically with the measurement beam and spatially separated into its p and s polarizations. The relative phase of the resulting temporal fringes is a linear function of the polarizer azimuthal angle P, and thus this component can be driven to its null position without iteration. Once at null, the azimuthal angle of the reflected, linearly polarized light is trivially determined from the relative amplitude of the fringes. Measurements made with this instrument on a native oxide film on a silicon wafer were in excellent agreement with those made with a traditional PCSA null ellipsometer. PMID- 19904329 TI - Algorithm for the identification of malfunctioning sensors in the control systems of segmented mirror telescopes. AB - The active control systems of segmented mirror telescopes are vulnerable to a malfunction of a few (or even one) of their segment edge sensors, the effects of which can propagate through the entire system and seriously compromise the overall telescope image quality. Since there are thousands of such sensors in the extremely large telescopes now under development, it is essential to develop fast and efficient algorithms that can identify bad sensors so that they can be removed from the control loop. Such algorithms are nontrivial; for example, a simple residual-to-the-fit test will often fail to identify a bad sensor. We propose an algorithm that can reliably identify a single bad sensor and we extend it to the more difficult case of multiple bad sensors. Somewhat surprisingly, the identification of a fixed number of bad sensors does not necessarily become more difficult as the telescope becomes larger and the number of sensors in the control system increases. PMID- 19904330 TI - Edge enhancement for in-phase focal modulation microscope. AB - In-phase focal modulation microscopy (IPFMM) with single photon excited fluorescence is presented. Optical transfer functions and images of thin and thick fluorescent edges in IPFMM are investigated. The results show that, compared with confocal microscopy, using IPFMM can result in a sharper image of the edge, and the edge gradient can be increased up to 75.4% and 58.9% for a thick edge and a thin edge, respectively. Signal level is also discussed, and the results show that, to obtain high transverse resolution with IPFMM, the normalized detector pinhole radius should not exceed 2.8. PMID- 19904331 TI - Optimal dark hole generation via two deformable mirrors with stroke minimization. AB - The past decade has seen a significant growth in research targeted at space-based observatories for imaging exosolar planets. The challenge is in designing an imaging system for high contrast. Even with a perfect coronagraph that modifies the point spread function to achieve high contrast, wavefront sensing and control is needed to correct the errors in the optics and generate a "dark hole." The high-contrast imaging laboratory at Princeton University is equipped with two Boston Micromachines Kilo-DMs. We review here an algorithm designed to achieve high contrast on both sides of the image plane while minimizing the stroke necessary from each deformable mirror (DM). This algorithm uses the first DM to correct for amplitude aberrations and uses the second DM to create a flat wavefront in the pupil plane. We then show the first results obtained at Princeton with this correction algorithm, and we demonstrate a symmetric dark hole in monochromatic light. PMID- 19904332 TI - Hybrid robust and fast algorithm for three-dimensional phase unwrapping. AB - We present a hybrid three-dimensional (3D) unwrapping algorithm that combines the strengths of two other fast and robust existing techniques. In particular, a branch-cut surface algorithm and a path-following method have been integrated in a symbiotic way, still keeping execution times within a range that permits their use in real-time applications that need a relatively fast solution to the problem. First, branch-cut surfaces are calculated, disregarding partial residue loops that end at the boundary of the 3D phase volume. These partial loops are then used to define a quality for each image voxel. Finally, unwrapping proceeds along a path determined by a minimum spanning tree (MST). The MST is built according to the quality of the voxels and avoids crossing the branch-cut surfaces determined at the first step. The resulting technique shows a higher robustness than any of the two methods used in isolation. On the one hand, the 3D MST algorithm benefits from the branch-cut surfaces, which endows it with a higher robustness to noise and open-ended wraps. On the other hand, incorrectly placed surfaces due to open loops at the boundaries in the branch-cut surface approach disappear. PMID- 19904333 TI - Systematic errors analysis for a large dynamic range aberrometer based on aberration theory. AB - In Ref. 1, it was demonstrated that the significant systematic errors of a type of large dynamic range aberrometer are strongly related to the power error (defocus) in the input wavefront. In this paper, a generalized theoretical analysis based on vector aberration theory is presented, and local shift errors of the SH spot pattern as a function of the lenslet position and the local wavefront tilt over the corresponding lenslet are derived. Three special cases, a spherical wavefront, a crossed cylindrical wavefront, and a cylindrical wavefront, are analyzed and the possibly affected Zernike terms in the wavefront reconstruction are investigated. The simulation and experimental results are illustrated to verify the theoretical predictions. PMID- 19904334 TI - Comparison of line-peak and line-scanning excitation in two-color laser-induced fluorescence thermometry of OH. AB - Two-line laser-induced-fluorescence (LIF) thermometry is commonly employed to generate instantaneous planar maps of temperature in unsteady flames. The use of line scanning to extract the ratio of integrated intensities is less common because it precludes instantaneous measurements. Recent advances in the energy output of high-speed, ultraviolet, optical parameter oscillators have made possible the rapid scanning of molecular rovibrational transitions and, hence, the potential to extract information on gas-phase temperatures. In the current study, two-line OH LIF thermometry is performed in a well-calibrated reacting flow for the purpose of comparing the relative accuracy of various line-pair selections from the literature and quantifying the differences between peak intensity and spectrally integrated line ratios. Investigated are the effects of collisional quenching, laser absorption, and the integration width for partial scanning of closely spaced lines on the measured temperatures. Data from excitation scans are compared with theoretical line shapes, and experimentally derived temperatures are compared with numerical predictions that were previously validated using coherent anti-Stokes-Raman scattering. Ratios of four pairs of transitions in the A2Sigma+<--X2Pi (1,0) band of OH are collected in an atmospheric-pressure, near-adiabatic hydrogen-air flame over a wide range of equivalence ratios--from 0.4 to 1.4. It is observed that measured temperatures based on the ratio of Q1(14)/Q1(5) transition lines result in the best accuracy and that line scanning improves the measurement accuracy by as much as threefold at low-equivalence-ratio, low-temperature conditions. These results provide a comprehensive analysis of the procedures required to ensure accurate two-line LIF measurements in reacting flows over a wide range of conditions. PMID- 19904335 TI - Determination of cell elasticity through hybrid ray optics and continuum mechanics modeling of cell deformation in the optical stretcher. AB - The optical stretcher is a dual-beam trap capable of stretching individual cells. Previous studies have used either ray- or wave-optical models to compute the optical pressure on the surface of a spherical cell. We have extended the ray optics model to account for focusing by the spherical interface and the effects of multiple internal reflections. Simulation results for red-blood cells (RBCs) show that internal reflections can lead to significant perturbation of the deformation, leading to a systematic error in the determination of cellular elasticity. Calibration studies show excellent agreement between the predicted and measured escape force, and RBC stiffness measurements are consistent with literature values. Measurements of the elasticity of murine osteogenic cells reveal that these cells are approximately 5.4 times stiffer than RBCs. PMID- 19904336 TI - Radiometric versus geometric, linear, and nonlinear vignetting coefficient. AB - We analyze the vignetting phenomenon both for optical systems with objects placed at finite distances and for systems with objects at infinity. Four of the possible definitions of the vignetting coefficient k, only two of them existing in the literature, are discussed. We propose two new definitions, i.e., a nonlinear geometric coefficient that is, in part, an analytical model of the vignetting characterization using optical software and a radiometric vignetting coefficient. The object space of each type of optical systems is studied first, defining its characteristic light circles and cones. Several simplifying assumptions are made for each of the two cases considered to derive analytical equations of the vignetting coefficient and thus to determine the best definition to be used. A geometric vignetting coefficient with two expressions, a linear classical and easy-to-use one and a nonlinear, that we propose for both types of systems is obtained. This nonlinear geometric vignetting coefficient proves to be more adequate in modeling the phenomenon, but it does not entirely fit the physical reality. We finally demonstrate that the radiometric vignetting coefficient we define and derive as a view factor for both types of optical systems is the most appropriate one. The half vignetting level, necessary in most optical design procedures to obtain a satisfactory illumination level in the image plane, is also discussed. PMID- 19904338 TI - Comparative study on the intracavity frequency-doubling 532 nm laser based on gray-tracking-resistant KTP and conventional KTP. AB - A comparative study of a frequency-doubling 532 nm laser based on gray-tracking resistant KTP (GTR-KTP) and conventional KTP is presented. The intracavity GTR KTP was proved to have better temperature characteristics than that of conventional KTP. Within the normalized output power variation range of 0.8-1.0, GTR-KTP has a temperature tolerance of 35 degrees C, broader than the 21 degrees C obtained with conventional KTP. Under the laser diode (LD) pump power of 180 W, the maximum average output power at 532 nm was 40.6 W for GTR-KTP at a repetition frequency of 10 kHz. In the case of conventional KTP, the maximum available LD pump power was limited to 150 W, with the corresponding maximum green average output power of 27.2 W. PMID- 19904337 TI - Real-time polarization-sensitive optical coherence tomography data processing with parallel computing. AB - With the increase of the A-line speed of optical coherence tomography (OCT) systems, real-time processing of acquired data has become a bottleneck. The shared-memory parallel computing technique is used to process OCT data in real time. The real-time processing power of a quad-core personal computer (PC) is analyzed. It is shown that the quad-core PC could provide real-time OCT data processing ability of more than 80 K A-lines per second. A real-time, fiber based, swept source polarization-sensitive OCT system with 20 K A-line speed is demonstrated with this technique. The real-time 2D and 3D polarization-sensitive imaging of chicken muscle and pig tendon is also demonstrated. PMID- 19904339 TI - Systematic error of a large dynamic range aberrometer. AB - Shack-Hartmann aberrometers are routinely used for measuring ocular aberrations. In one configuration, the intermediate images of the Shack-Hartmann spots formed by the lenslet array are relayed by an imaging lens onto a sensor. A systematic introduction of spherical aberration that is strongly related to the power error (defocus) of the incident wavefront is observed in this configuration. We found that the largest component of this error is induced by the pupil aberration of the imaging relay lens. Some simulations and experimental results are demonstrated. PMID- 19904340 TI - Measured turbulent mixing in a small-scale circuit breaker model. AB - The performance of high voltage gas circuit breakers depends on the temperature distribution of hot gas or plasma from the arc zone mixed with cold gas that is present, for example, in the exhausts and mixing volume. Understanding the details of the mixing process is imperative to estimate the temperature distribution within the entire breaker volume. Design studies rely on computational fluid dynamics (CFD) simulations to search for the best way to achieve satisfactory mixing. One key uncertainty in the CFD simulations is the role of turbulence in this process and how to properly account for it. To gain knowledge of the mixing process between hot and cold gases, we have constructed a simplified breaker geometry that is flexible and accessible to diagnostics. Apart from standard measurements of current and arc voltage, we measure pressure in the arc zone and the mixing volume. Further, the mixing volume is specially designed to be transparent, allowing us to make shadowgraphy measurements of the turbulent mixing during and after the arcing phase. We report on experiments performed in air at atmospheric pressure. PMID- 19904341 TI - Active hexagonally segmented mirror to investigate new optical phasing technologies for segmented telescopes. AB - The primary mirror of the future European Extremely Large Telescope will be equipped with 984 hexagonal segments. The alignment of the segments in piston, tip, and tilt within a few nanometers requires an optical phasing sensor. A test bench has been designed to study four different optical phasing sensor technologies. The core element of the test bench is an active segmented mirror composed of 61 flat hexagonal segments with a size of 17 mm side to side. Each of them can be controlled in piston, tip, and tilt by three piezoactuators with a precision better than 1 nm. The context of this development, the requirements, the design, and the integration of this system are explained. The first results on the final precision obtained in closed-loop control are also presented. PMID- 19904345 TI - Electrochemistry at carbon nanotubes: perspective and issues. AB - Electrochemistry at carbon nanotubes (CNTs) is a large and growing field, but one in which there is still uncertainty about the fundamental activity of CNTs as electrode materials. On the one hand, there are many reports which focus on the favourable electrochemical properties of CNT electrodes, such as enhanced detection sensitivity, electrocatalytic effects and reduced fouling. On the other hand, other studies suggest that CNTs may be no more electroactive than graphitic powder. Furthermore, it has been proposed that the catalytic nanoparticles from which CNTs are formed may dominate the electrochemical characteristics in some instances. A considerable body of the literature presumes that the CNT sidewall is inert and that edge-plane-graphite-like open ends and defect sites are responsible for the electron transfer activity observed. In contrast, studies of well characterised single-walled nanotube (SWNT) electrodes, either as individual tubes or as two-dimensional networks, suggest sidewall activity. This review highlights how the various discrepancies in CNT electrochemistry may have arisen, by taking a historical view of the field and identifying crucial issues that still need to be solved. When assessing the behaviour of CNT electrodes, it is vitally important that careful consideration is given to the type of CNT used (SWNT or multi-walled), the quality of the material (presence of impurities), the effect of chemical processing steps in the fabrication of electrodes and the experimental arrangements adopted. Understanding these key features is an essential requirement to develop a fundamental understanding of CNT electrochemistry, to allow a wide range of electroanalytical applications, and to move the field forward rationally. As part of this process, high resolution electrochemical and electrical imaging techniques are expected to play a significant role in the future, as well as theoretical developments which examine the fundamentals of electron transfer at different types of CNTs and their characteristic surface sites. PMID- 19904346 TI - Raman spectroscopy of strained single-walled carbon nanotubes. AB - Due to remarkable rolling structure and distinct rolling direction, the chirality dependent Raman spectra of single-walled carbon nanotubes (SWNTs) show two characteristic features: the radial breathing mode (RBM) and the G-band. Rich information about SWNTs presented by these Raman features makes Raman spectroscopy a general and common tool for characterizing structures and properties of SWNTs and their changes. When exerted by external forces, the geometrical structures of SWNTs will change, which further affects the electronic structures and phonon properties of SWNTs. In this article, emphasis is given to how Raman frequency and resonant-Raman intensity evolve under distinct strains, including uniaxial strain, torsional strain, radial deformation and bending deformation. It is found that depending on different structural variations, Raman spectra of SWNTs have different responses to each strain, showing that resonant Raman spectroscopy is a suitable tool to characterize and study strains in SWNTs. PMID- 19904347 TI - Porous graphenes: two-dimensional polymer synthesis with atomic precision. AB - We demonstrate, by surface-assisted coupling of specifically designed molecular building blocks, the fabrication of regular two-dimensional polyphenylene networks with single-atom wide pores and sub-nanometer periodicity. PMID- 19904348 TI - Magnetic chiral ionic liquids derived from amino acids. AB - Novel room temperature magnetic chiral ionic liquids derived from amino acids were synthesized and their magnetic properties as well as chiral discrimination abilities were investigated. PMID- 19904350 TI - Synthesis, characterization, and self-assembled nanofibers of carbohydrate functionalized mono- and di(2,2':6',2''-terpyridinyl)arenes. AB - Self-assembly of novel linear and branched carbohydrate-functionalized mono- and di(2,2':6',2''-terpyridinyl)arenes afforded access to a series of twisted self organized nanofibers. PMID- 19904349 TI - Synthesis of homogeneous antifreeze glycopeptides via a ligation-desulfurisation strategy. AB - Homogeneous glycopeptide analogues of fish antifreeze glycoproteins of discrete oligomeric length have been synthesised using a native chemical ligation desulfurisation strategy. PMID- 19904351 TI - A triple-stranded helicate and mesocate from the same metal and ligand. AB - A pair of triple-stranded helicates and mesocates were, for the first time, isolated from the same reaction of a novel alpha-free bis(dipyrromethene) ligand with either Co(3+) or Fe(3+). PMID- 19904352 TI - Synthesis and structure of a magnesium-amidoborane complex and its role in catalytic formation of a new bis-aminoborane ligand. AB - A synthetic route to a magnesium-amidoborane complex and its role in the catalytic conversion of a substituted ammonia-borane RNH(2)BH(3) into HB(NHR)(2) is discussed. PMID- 19904353 TI - Controlled growth of novel hyper-branched nanostructures in nanoporous alumina membrane. AB - This paper proposes a novel approach to fabricate hyper-branched anodic aluminium oxide (AAO) nanostructures with different branches on the vertically-aligned trunk and at the trunk terminal. Silver nanowires with different dimensional and multifunctional complexity have been prepared from this hyper-branched AAO template by varying the electrodeposition time. These kinds of novel nanostructure may be used to build blocks for nanoelectronic and nanophotonic devices. PMID- 19904354 TI - Assembling chirality into magnetic nanowires: cyano-bridged iron(III)-nickel(II) chains exhibiting slow magnetization relaxation and ferroelectricity. AB - The assembly of chirality into magnetic nanowires enables the formation of two new cyano-bridged heterobimetallic 3,2-chain enantiomeric complexes, representing the first example of multiferroic compounds bearing both slow magnetization relaxation and ferroelectricity. PMID- 19904355 TI - Reversible solid-gas chemical equilibrium between a 0-periodic deformable molecular tecton and a 3-periodic coordination architecture. AB - The reversible crystal transformation between a hexagonal (R3[combining macron]) porous coordination architecture [Cu(L)(2)](n) (L = 3-(pyridin-4-yl)-5-(pyrazin-2 yl)-1H-1,2,4-triazole), and a monoclinic (C2/c) molecular tecton [Cu(L)(2)(H(2)O)], was chemically implemented and structurally interpreted. PMID- 19904356 TI - A DNA-based electrochemical strategy for label-free monitoring the activity and inhibition of protein kinase. AB - A novel label-free electrochemical strategy for monitoring the activity and inhibition of protein kinase is developed, based on the linkage between the phosphorylated peptide and DNA functionalized Au nanoparticles (DNA-AuNPs) by Zr(4+) and the chronocoulometric response of [Ru(NH(3))(6)](3+) absorbed on the DNA-AuNPs. PMID- 19904357 TI - Protein surface recognition by dendritic ruthenium(II) tris(bipyridine) complexes. AB - We report protein surface binding of dendritic ruthenium(ii) tris(bipyiridine) complexes to alpha-chymotrypsin, resulting in a 1 : 1 and 1 : 2 protein complex formation as well as inhibition of the enzyme activity. PMID- 19904358 TI - Multifunctional perfluorocarbon nanoemulsions for (19)F-based magnetic resonance and near-infrared optical imaging of dendritic cells. AB - A novel type of bimodal imaging nanoprobe based on (19)F-based magnetic resonance imaging and near-infrared optical imaging has been synthesized and applied for the labeling and imaging of dendritic cells both in vitro and in vivo. PMID- 19904359 TI - N-type cascade electron transfer along an oxidative gradient. AB - N-type cascade electron transfer was achieved for the first time along an oxidative gradient in the triad of ferrocene-perylene tetracarboxylic bisimide [60]fullerene, providing a promising new strategy of supramolecular architecture for solar energy conversion. PMID- 19904360 TI - Design of one-to-one recognition triple Au nanoparticles DNA probe and its application in the electrochemical DNA biosensor. AB - A novel and highly sensitive biosensor for sequence specific DNA detection was constructed based on a strategy of one-to-one recognition reaction between Tri AuNPs DNA probe signal amplification unit and target DNA. PMID- 19904361 TI - Synthesis, photophysical properties and color tuning of highly fluorescent 9,10 disubstituted-2,3,6,7-tetraphenylanthracene. AB - We report the synthesis of 9,10-disubstituted-2,3,6,7-tetraphenylanthracenes, of which the photophysical properties are altered significantly by the 9,10 functionalities; the emission wavelengths range from 410 to 610 nm with outstanding quantum yields for most fluorophores. PMID- 19904362 TI - Non-centrosymmetric behavior of a clay film ion-exchanged with chiral metal complexes. AB - SHG measurements on a highly transparent clay film ion-exchanged with chiral metal complexes revealed that the mono-molecular layer of the chiral complexes in an interlayer space acquired a non-centrosymmetric character. PMID- 19904363 TI - ZinCast-1: a photochemically active chelator for Zn(2+). AB - Two strategies were applied to the synthesis of ZinCast-1, a nitrobenzhydrol based caged complex that upon photolysis exhibits a nearly 400-fold difference in binding for Zn(2+). PMID- 19904364 TI - Regioselective synthesis of halohydrin esters from epoxides: reaction with acyl halides and rhodium-catalyzed three-component coupling reaction with alkyl halides and carbon monoxide. AB - Regioselective synthesis of halohydrin esters was achieved by (1) the reaction of acyl halides with epoxides and (2) the rhodium-catalyzed three-component coupling reaction of alkyl halides, carbon monoxide, and epoxides. PMID- 19904365 TI - A novel click chitooligosaccharide for hydrophilic interaction liquid chromatography. AB - A novel chitooligosaccharide stationary phase for hydrophilic interaction liquid chromatography (HILIC) was developed via click chemistry and showed great HILIC characteristics on separation of polar compounds and enrichment of glycopeptides. PMID- 19904366 TI - Enzymatic synthesis of UDP-GlcNAc/UDP-GalNAc analogs using N-acetylglucosamine 1 phosphate uridyltransferase (GlmU). AB - Reports the generation of a library composed of UDP-GlcNAc/UDP-GalNAc and investigates the substrate specificity of Escherichia coli GlcNAc-1-P uridyltransferase GlmU. PMID- 19904367 TI - Borylene-based functionalization of Pt-alkynyl complexes by photochemical borylene transfer from [(OC)(5)Cr=BN(SiMe(3))(2)]. AB - Photochemical borylene transfer from [(OC)(5)Cr=BN(SiMe(3))(2)] to the platinum sigma-alkynyl complex [Cl(PMe(3))(2)Pt-C[triple bond]CPh] gave facile access to the first transition metal-borirene complex. The title compound has been fully characterized in solution and by X-ray crystallography. PMID- 19904368 TI - Fluorescent protein red Kaede chromophore; one-step, high-yield synthesis and potential application for solar cells. AB - The one-step condensation between the Green Fluorescence Protein (GFP) chromophore and 1H-imidazole-5-carbaldehyde catalyzed by ZnCl(2) proved to be a facile method to synthesize the red Kaede chromophore and its derivatives (1a-d, 2, and 3 > 70%), among which 1a then demonstrated its potential as a dye in solar cell, with a moderate conversion efficiency (eta) of 3.04%. PMID- 19904369 TI - Solid-phase synthesis of protected alpha-amino phosphonic acid oligomers. AB - By establishing both a highly efficient phosphonamidate formation and a RuCp catalyzed cleavage of an allyl linker, the solid-phase synthesis of Fmoc (GlyP(OBn))(6)-OH/DIEA, a protected form of a new type of unnatural peptide alpha amino phosphonic acid oligomer (APO), has been realized. PMID- 19904370 TI - Asymmetric Au(i)-catalyzed synthesis of bicyclo[4.1.0]heptene derivatives via a cycloisomerization process of 1,6-enynes. AB - The enantioselective asymmetric gold-catalyzed cycloisomerization reactions of heteroatom tethered 1,6-enynes are conducted in the presence of a chiral cationic Au(i) catalyst ((R)-4-MeO-3,5-(t-Bu)(2)-MeOBIPHEP-(AuCl)(2)/AgOTf system) in toluene under mild conditions and lead to functionalized bicyclo[4.1.0]heptene derivatives in excellent enantiomeric excesses ranging from 90-98%. PMID- 19904371 TI - Umpolung of halide reactivity: efficient (diacetoxyiodo)benzene-mediated electrophilic alpha-halogenation of 1,3-dicarbonyl compounds. AB - An efficient high-yielding (diacetoxyiodo)benzene-mediated alpha-halogenation of 1,3-dicarbonyl compounds utilising titanium tetrahalides as the halide source has been developed. PMID- 19904373 TI - Salt, pH and thermoresponsive supramolecular hydrogel of N-(4-n tetradecyloxybenzoyl)-L-carnosine. AB - A carnosine based amphiphilic hydrogelator has been developed which efficiently gelates water and exhibits salt, pH and thermoresponsive gelation properties. PMID- 19904372 TI - Merging chiral organocatalysts: enantio- and diastereoselective direct vinylogous Mannich reaction of alkylimines. AB - An enantio- and diastereoselective direct vinylogous Mannich reaction of alpha,alpha-dicyanoolefins and N-sulfonyl alkylimines has been developed by the catalysis of a new family of bifunctional organocatalysts merging chiral BINOL and 9-amino-9-deoxyepi-cinchona alkaloid skeletons, from which chiral beta-, delta- or gamma-amino compounds could be efficiently derived. PMID- 19904374 TI - Sulfonic acid functionalized crystal-like mesoporous benzene-silica as a remarkable water-tolerant catalyst. AB - Here we report that sulfonated crystal-like benzene-silica is much more robust and active than conventionally used periodic mesoporous silica for catalyzing aqueous organic reactions. PMID- 19904375 TI - A nanophase segregated mesophase morphology in self-organized novel disc-rod oligomesogens. AB - A nanophase segregated layered phase (SmA) with alternating calamitic and discotic layers was observed for the first time in novel linked disc-rod oligomesogens, containing six cyanobiphenyl moieties radially-attached to a central tricycloquinazoline discotic heteroaromatic core via flexible alkyl spacers. PMID- 19904376 TI - First time-resolved EPR observation of Nafion photochemistry. AB - Time-resolved EPR spectra of UV-irradiated Nafion reveal the formation of spin polarized excited triplet states and allow the detection of photoinduced triplet triplet energy transfer processes through hydrogen bonds between water and sulfonic acid groups. PMID- 19904377 TI - Iodine-mediated cyclization of N-thioacyl-1-(2-pyridyl)-1,2-aminoalcohols and their subsequent condensation leading to the formation of novel bis(1-imidazo[1,5 a]pyridyl)arylmethanes. AB - The treatment of N-thioacyl-1-(2-pyridyl)-1,2-aminoalcohols with iodine and pyridine in THF at room temperature for 30 min leads to the formation of bis(1 imidazo[1,5-a]pyridyl)arylmethanes as green solids in good to high yields. PMID- 19904378 TI - Photo-bleaching immunity encoded photonic suspension array for label-free multiplex analysis. AB - A new type of suspension array for multiplex analysis has been proposed by using polarized luminescent ratios of dichroic dyes as the encoding elements, which provided highly stable codes and advantages of label-free detection. PMID- 19904379 TI - Development of 2,6-carboxy-substituted boron dipyrromethene (BODIPY) as a novel scaffold of ratiometric fluorescent probes for live cell imaging. AB - Ratiometric fluorescent probes based on boron dipyrromethene (BODIPY) were developed based on a novel design strategy, in which a change of the electron withdrawing character of the 2,6-substituents resulting from reaction with a target molecule generates a fluorescence wavelength change. PMID- 19904380 TI - Towards a tunable and switchable water adhesion on a TiO(2) nanotube film with patterned wettability. AB - Tunable water adhesion was realized on a TiO(2) nanotube film with patterned wettability formed via selective illumination through a mask. Meanwhile, the adhesion can be switched between sliding superhydrophobicity and sticky superhydrophobicity by masked illumination and heat annealing. PMID- 19904381 TI - In situ, real-time tracking of cell wall topography and nanomechanics of antimycobacterial drugs treated Mycobacterium JLS using atomic force microscopy. AB - Two antimycobacterial drugs ethambutol (EMB) and isoniazid (INH) were found to result in decreases of biomechanical properties including adhesion force, cell wall spring constant, and Young's modulus in Mycobacterium sp. strain JLS (M. JLS). PMID- 19904382 TI - A F-bridged Mn(II) molecular square. AB - We report on a tetranuclear Mn(ii) complex possessing a F-bridged molecular square topology directed by 1,10-phenanthroline ligands. PMID- 19904383 TI - Thermosensitive hydrogels composed of cyclodextrin pseudorotaxanes. Role of [3]pseudorotaxane in the gel formation. AB - Pseudorotaxanes composed of an alkylpyridinium and alpha-cyclodextrin (alpha-CD) form supramolecular hydrogels which show sol-gel transitions at 7-67 degrees C depending on the type and amount of the guest compound. PMID- 19904384 TI - Visual and colorimetric detection of Hg(2+) by cloud point extraction with functionalized gold nanoparticles as a probe. AB - Association with Hg(2+) enhances the hydrophobicity and triggers the cloud point extraction of approximately 4 nm-diameter gold nanoparticle probes functionalized with mercaptopropionic acid and homocystine, which results in the color change of the TX-114-rich phase from colorless to red, and therefore provides a novel approach for visual and colorimetric detection of Hg(2+) with ultrahigh sensitivity and selectivity. PMID- 19904385 TI - Biological screening of a diverse set of AI-2 analogues in Vibrio harveyi suggests that receptors which are involved in synergistic agonism of AI-2 and analogues are promiscuous. AB - C1-alkyl AI-2 analogues do not induce bioluminescence in V. harveyi on their own but enhance the bioluminescence induced by AI-2 in a synergistic fashion. A new facile synthesis of AI-2 facilitates the synthesis of a diverse set of AI-2 analogues and biological screening suggests that receptors that are involved in the synergistic bioluminescence production in V. harveyi are promiscuous. PMID- 19904386 TI - Enhanced resistance of DNA nanostructures to enzymatic digestion. AB - The ability of nucleases to perform their catalytic functions depends on the sequence and structural features of target DNA substrates. Due to their size and shape, several DNA tetrahedra are resistant to the action of specific and non specific nucleases. Such enhanced stability is a key requirement for DNA nanostructures to be useful as delivery vehicles. PMID- 19904387 TI - A new versatile solvatochromic amino-macrocycle. From metal ions to cell sensing in solution and in the solid state. AB - A new fluorescent NBD-polyaza-macrocycle sensor (L) was synthesized. The coordination of Cu(ii) and Zn(ii) in acetonitrile switches on the fluorescence with different emission wavelengths. Cu(ii) complexes showed solid-state fluorescence. Both L and Cu-complex interact with human cell line (U937) highlighting the cell membrane by fluorescence microscopy. PMID- 19904388 TI - A novel composite hierarchical hollow structure: one-pot synthesis and magnetic properties of W(18)O(49)-WO(2) hollow nanourchins. AB - A novel composite hierarchical hollow structure is reported. The as-prepared products consist of discrete WO(2) hollow core spheres with W(18)O(49) nanorod shells ("hollow urchins"). The products showed unusual magnetic behavior. PMID- 19904389 TI - One-pot cross-coupling of N-acyl N,O-acetals with alpha,beta-unsaturated compounds. AB - The synergistic action of BF(3).OEt(2) and SmI(2) allowed a series of intermolecular cross-couplings of readily available N-acyl N,O-acetals with alpha,beta-unsaturated compounds to be performed in high yields, which was applied to the stereoselective synthesis of pyrrolizidine alkaloid (+) xenovenine. PMID- 19904390 TI - Ultrathin single crystal Pt nanowires grown on N-doped carbon nanotubes. AB - Ultrathin single crystal platinum nanowires with diameters of approximately 2.5 nm and lengths up to 100 nm were synthesized on nitrogen-doped CNTs (N-CNTs) via a straightforward wet-chemical method in environmentally friendly water solution at room temperature, without using any stabilizing agent. These ultrathin nanowires and their composites with N-CNTs hold potential for a wide range of applications. PMID- 19904396 TI - IP or no IP: that is the question. PMID- 19904397 TI - Toward one-step point-of-care immunodiagnostics using capillary-driven microfluidics and PDMS substrates. AB - Point-of-care diagnostics will strongly benefit from miniaturization based on microfluidics because microfluidics integrate functions that can together preserve valuable samples and reagents, increase sensitivity of a test, and accelerate mass transport limited reactions. But a main challenge is to incorporate reagents into microfluidics and to make microfluidics simple to use. Here, we integrate microfluidic functional elements, some of which were developed earlier, and reagents such as detection antibodies (dAbs), capture antibodies (cAbs) and analyte molecules for making one-step immunoassays: the integrated device only requires the addition of sample to trigger a cascade of events powered by capillary forces for effecting a sandwich immunoassay that is read using a fluorescence microscope. The microfluidic elements comprise a sample collector, delay valves, flow resistors, a deposition zone for dAbs, a reaction chamber sealed with a polydimethylsiloxane (PDMS) substrate, and a capillary pump and vents. Parameters for depositing 3.6 nL of a solution of dAb on the chip using an inkjet are optimized and the PDMS substrate is patterned with analytes, which provide a positive control, and cAbs. Various storage conditions of the patterned PDMS are investigated for up to 6 months revealing that storage with a desiccant preserved at least 51% of the activity of the cAbs. C-reactive protein (CRP), a general inflammation and cardiac marker, is detected using this one-step chip using only 5 microL of human serum by measuring fluorescent signals from 30 x 100 microm(2) areas of the PDMS substrate in the wet reaction chamber. The one step chip can detect CRP at a concentration of 10 ng mL(-1) in less than 3 min and below 1 ng mL(-1) within 14 min. The work presented here may spur the adoption of fluorescence immunoassays using capillary driven microfluidics and PDMS substrates for point-of-care diagnostics. PMID- 19904398 TI - Tunable shrink-induced honeycomb microwell arrays for uniform embryoid bodies. AB - Embryoid body (EB) formation closely recapitulates early embryonic development with respect to lineage commitment. Because it is greatly affected by cell-cell and cell-substrate interactions, the ability to control the initial number of cells in the aggregates and to provide an appropriate substrate are crucial parameters for uniform EB formation. Here we report of an ultra-rapid fabrication and culture method utilizing a laser-jet printer to generate closely arrayed honeycomb microwells of tunable sizes for the induction of uniform EBs from single cell suspension. By printing various microwell patterns onto pre-stressed polystyrene sheets, and through heat induced shrinking, high aspect micromolds are generated. Notably, we achieve rounded bottom polydimethylsiloxane (PDMS) wells not easily achievable with standard microfabrication methods, but critical to achieve spherical EBs. Furthermore, by simply controlling the size of the microwells and the concentration of the cell suspension we can control the initial size of the cell aggregate, thus influencing lineage commitment. In addition, these microwells are easily adaptable and scalable to most standard well plates and easily integrated into commercial liquid handling systems to provide an inexpensive and easy high throughput compound screening platform. PMID- 19904400 TI - Continuous particle separation in a microfluidic channel via standing surface acoustic waves (SSAW). AB - This work introduces a method of continuous particle separation through standing surface acoustic wave (SSAW)-induced acoustophoresis in a microfluidic channel. Using this SSAW-based method, particles in a continuous laminar flow can be separated based on their volume, density and compressibility. In this work, a mixture of particles of equal density but dissimilar volumes was injected into a microchannel through two side inlets, sandwiching a deionized water sheath flow injected through a central inlet. A one-dimensional SSAW generated by two parallel interdigital transducers (IDTs) was established across the channel, with the channel spanning a single SSAW pressure node located at the channel center. Application of the SSAW induced larger axial acoustic forces on the particles of larger volume, repositioning them closer to the wave pressure node at the center of the channel. Thus particles were laterally moved to different regions of the channel cross-section based on particle volume. The particle separation method presented here is simple and versatile, capable of separating virtually all kinds of particles (regardless of charge/polarization or optical properties) with high separation efficiency and low power consumption. PMID- 19904399 TI - Multiplex pressure measurement in microsystems using volume displacement of particle suspensions. AB - We demonstrate a simple image-based method to measure pressure in microsystems using volume displacement of fluorescent particle suspensions. These micro pressure-sensors are composed of two layers with a poly(dimethylsiloxane) (PDMS) membrane in between: the flow layer includes a flow channel and the sensor layer contains a detection channel filled with suspensions of fluorescent particles. The pressure increase in the flow channel deflects the membrane, and this membrane deformation can be quantified by measuring the cross-sectional areas at specific focal planes. These simple sensors have the advantage that a broad sensing-range can be achieved by tuning the mechanical property and the geometry of the membrane during design and fabrication, and even simpler by tuning the focal plane or the pressure of a reference chamber while in operation. We also demonstrate here a pressure transduction scheme coupled with the image-based sensing method as a multiplex pressure measurement tool for simultaneously detecting pressures in multiple locations in a microsystem. Overall, the image based pressure sensing method has high precision when operated in both direct and remote detection modes. Compared to conventional mechanical methods of pressure detection, this technique is inexpensive because it does not require complex off chip equipment to quantify the pressure-dependent membrane deformation. In addition, the image analysis using the software code developed here is fast, and it generates data that are simple to interpret. PMID- 19904401 TI - Label-free and highly sensitive biomolecular detection using SERS and electrokinetic preconcentration. AB - In this paper, we present a method combining surface-enhanced Raman scattering (SERS) spectroscopy to detect biomolecules in a label-free way with an electrokinetic preconcentration technique (electrophoresis) to amplify biomolecular signals at low concentrations. A constant electric field is applied to charged biomolecules in solution, attracting them to an oppositely charged electrode, which is also used as a SERS substrate. Within 5 min, we observed that the SERS signal of 10 fM adenine was amplified to the level of the signal of non preconcentrated 1 microM adenine (sensitivity improvement by 8 orders of magnitude) and the method was effective over a wide range of concentrations (10 fM to 1 microM). The signals were further amplified under stronger electric field and longer application: The increase of the signal intensity was observed to be 51 times at -0.6 V cm(-1) after 25 min. The effectiveness of this method allows the creation of label-free, target-specific, and highly sensitive monitoring applications. PMID- 19904402 TI - Rapid automated cell quantification on HIV microfluidic devices. AB - Lab-chip device analysis often requires high throughput quantification of fluorescent cell images, obtained under different conditions of fluorescent intensity, illumination, focal depth, and optical magnification. Many laboratories still use manual counting--a tedious, expensive process prone to inter-observer variability. The manual counting process can be automated for fast and precise data gathering and reduced manual bias. We present a method to segment and count cells in microfluidic chips that are labeled with a single stain, or multiple stains, using image analysis techniques in Matlab and discuss its advantages over manual counting. Microfluidic based cell capturing devices for HIV monitoring were used to validate our method. Captured CD4(+) CD3(+) T lymphocytes were stained with DAPI, AF488-anti CD4, and AF647-anti CD3 for cell identification. Altogether 4788 (76 x 3 x 21) gray color images were obtained from devices using discarded 10 HIV infected patient whole blood samples (21 devices). We observed that the automatic method performs similarly to manual counting for a small number of cells. However, automated counting is more accurate and more than 100 times faster than manual counting for multiple-color stained cells, especially when large numbers of cells need to be quantified (>500 cells). The algorithm is fully automatic for subsequent microscope images that cover the full device area. It accounts for problems that generally occur in fluorescent lab-chip cell images such as: uneven background, overlapping cell images and cell detection with multiple stains. This method can be used in laboratories to save time and effort, and to increase cell counting accuracy of lab-chip devices for various applications, such as circulating tumor cell detection, cell detection in biosensors, and HIV monitoring devices, i.e. CD4 counts. PMID- 19904403 TI - Single cell electric impedance topography: mapping membrane capacitance. AB - Single-cell electric impedance topography (sceTopo), a technique introduced here, maps the spatial distribution of capacitance (i.e. displacement current) associated with the membranes of isolated, living cells. Cells were positioned in the center of a circular recording chamber surrounded by eight electrodes. Electrodes were evenly distributed on the periphery of the recording chamber. Electric impedance measured between adjacent electrode pairs (10 kHz-5 MHz) was used to construct topographical maps of the spatial distribution of membrane capacitance. Xenopus Oocytes were used as a model cell to develop sceTopo because these cells consist of two visually distinguishable hemispheres, each with distinct membrane composition and structure. Results showed significant differences in the imaginary component of the impedance between the two oocyte hemispheres. In addition, the same circumferential array was used to map the size of the extracellular electrical shunt path around the cell, providing a means to estimate the location and shape of the cell in the recording chamber. PMID- 19904404 TI - Three-dimensional cell culture array using magnetic force-based cell patterning for analysis of invasive capacity of BALB/3T3/v-src. AB - A three-dimensional (3D) cell culture system has been fabricated using a magnetic force based cell patterning method, demonstrating a facile approach for the analysis of invasive capacity of BALB/3T3/v-src using an magnetic force and magnetite nanoparticles. The 3D cell patterning was performed using an external magnetic force and a pin holder, which enables the assembly of the magnetically labeled cells on the collagen gel-coated surface as array-like cell patterns, resulting in the development of a 3D in vitro culture model. The cells embedded in type I collagen showed a compacted, spheroid like configuration at each spot, and distinct, accelerated cell growth was observed in cancer model cells compared with the control cells. The developed 3D cell culture array was applied to the susceptibility assay of the GM6001 matrix metalloproteinase (MMP) inhibitor, a collagenase inhibitor; a distinct suppression of cell proliferation was observed, while little change was observed in 2D. The developed 3D cell culture array system is useful to assess the effects of pharmacologic and/or microenvironmental influences on tumor cell invasion. PMID- 19904405 TI - Lipase-catalyzed synthesis of isoamyl acetate in an ionic liquid/n-heptane two phase system at the microreactor scale. AB - A continuously operated psi-shaped microreactor was used for lipase-catalyzed synthesis of isoamyl acetate in the 1-butyl-3-methylpyridinium dicyanamide/n heptane two-phase system. The chosen solvent system with dissolved Candida antarctica lipase B, which was attached to the ionic liquid/n-heptane interfacial area due to its amphiphilic properties, was shown to be highly efficient and enabled simultaneous esterification and product removal. At preliminarily selected conditions regarding the type of acyl donor, its molar ratio to alcohol and enzyme concentration, 48.4 g m(-3) s(-1) of isoamyl acetate was produced, which was almost three-fold better as compared to the intensely mixed batch process. This was mainly a consequence of efficient reaction-diffusion dynamics in the microchannel system, where the developed flow pattern comprising of intense emulsification provided a large interfacial area for the reaction and simultaneous product extraction. PMID- 19904406 TI - A PMMA microfluidic droplet platform for in vitro protein expression using crude E. coli S30 extract. AB - Droplet based microfluidics are promising new tools for biological and chemical assays. In this paper, a high throughput and high sensitivity microfluidic droplet platform is described for in vitro protein expression using crude Escherichia coli S30 extract. A flow-focusing polymethylmethacrylate (PMMA) microchip was designed and integrated with different functions involving droplet generation, storage, separation and detection. The material used for the chip is superior to the previously tested polydimethylsiloxane (PDMS) due to its mechanical and chemical properties. Droplet formation characteristics such as size and generation rate are investigated systematically. The effect of surfactants Abil EM90 and Span80 in the oil phase on droplet formation and optical detection is also studied. The performance of the system is demonstrated by the high throughput and stable droplet generation and ultralow detection limit. The robustness of the system is also demonstrated by the successful synthesis of a green fluorescent protein (GFP) using E. coli S30 extract as a source of RNA translation reagents. PMID- 19904407 TI - Hands-free sample preparation platform for nucleic acid analysis. AB - A Lab-On-Chip system with an instrument is presented which is capable of performing total sample preparation and automated extraction of nucleic acid from human cell samples fixed in a methanol based solution. The target application is extraction of mRNA from cervical liquid based cytology specimens for detection of transformed HPV-infections. The device accepts 3 ml of sample and performs the extraction in a disposable polymer chip of credit card size. All necessary reagents for cell lysis, washing, and elution are stored on-chip and the extraction is performed in two filter stages; one for cell pre-concentration and the other for nucleic acid capture. Tests performed using cancer cell lines and cervical liquid based cytology specimens confirm the extraction of HPV-mRNA by the system. PMID- 19904408 TI - A microwave interferometric system for simultaneous actuation and detection of single biological cells. AB - In biomedical applications ranging from the study of pathogen invasion to drug efficacy assays, there is a growing need to develop minimally invasive techniques for single-cell analysis. This has inspired researchers to develop optical, electrical, microelectromechanical and microfluidic devices for exploring phenomena at the single-cell level. In this work, we demonstrate an electrical approach for single-cell analysis wherein a 1.6 GHz microwave interferometer detects the capacitance changes (DeltaC) produced by single cells flowing past a coplanar interdigitated electrode pair. The experimental and simulated capacitance changes generated by yeast cells are in close agreement. By using the capacitance changes of uniform polystyrene spheres (diameter = 5.7 microm) for calibration purposes, we demonstrate a 0.65 aF sensitivity in a 10 ms response time. Using an RC circuit, a low frequency sinusoidal potential is simultaneously superimposed on the electrode pair to generate a dielectrophoretic force that translates cells. Specifically, when yeast cells suspended in a solution of 90 ppm NaCl in deionized water are exposed to 10 kHz and 3 MHz potentials (ranging from 1-3 V(pp)), they experience negative and positive dielectrophoresis, respectively. The corresponding changes in cell elevation above the interdigitated electrodes are detected using the asymmetry of the capacitance signature produced by the cell. Cell elevation changes can be detected in less than 80 ms. The minimum detectable change in elevation is estimated to be 0.22 microm. This approach will have applications in rapid single-cell dielectrophoretic analysis, and may also prove useful in conjunction with impedance spectroscopy. PMID- 19904409 TI - Micro-fluidic actuation using magnetic artificial cilia. AB - We demonstrate advanced fluid manipulations using magnetic polymeric artificial cilia on the walls of a microfluidic channel. In nature, cilia are little hairs covering the surface of micro-organisms which enable them to manipulate a fluid on the micro-scale. The asymmetric movement of natural cilia is crucial to obtain a net fluid flow. We have developed a ferromagnetic polymer made from iron nanoparticles and polydimethylsiloxane, and describe a process that can structure the material into high aspect ratio lying artificial cilia with a length of 300 microm. These artificial cilia were actuated with a homogeneous rotating magnetic field (micro(0)H < 50 mT) generated with a compact external electromagnet. An asymmetric movement involving torsion could be created when the cilia were provided with a remanent magnetisation perpendicular to the plane of rotation of the magnetic field vector. The artificial cilia could be actuated in fluid up to a frequency of approximately 50 Hz. In an aqueous solution in a microfluidic chamber we were able to generate rotational as well as translational fluid movements with fluid velocities up to approximately 0.5 mm s(-1). PMID- 19904410 TI - Passive microfluidic pumping using coupled capillary/evaporation effects. AB - Controlled pumping of fluids through microfluidic networks is a critical unit operation ubiquitous to lab-on-a-chip applications. Although there have been a number of studies involving the creation of passive flows within lab-on-a-chip devices, none has shown the ability to create temporally stable flows for periods longer than several minutes. Here a passive pumping approach is presented in which a large pressure differential arising from a small, curved meniscus situated along the bottom corners of an outlet reservoir serves to drive fluid through a microfluidic network. The system quickly reaches steady-state and is able to provide precise volumetric flow rates for periods lasting over an hour. A two-step mathematical model provides accurate predictions of fluid and mass transport dynamics in these devices, as validated by particle tracking in laboratory systems. Precise flow rates spanning an order of magnitude are accomplished via control of the microchannel and outlet reservoir dimensions. This flow mechanism has the potential to be applied to many micro-total analytical system devices that utilize pressure-driven flow; as an illustrative example, the pumping technique is applied for the passive generation of temporally stable chemical gradients. PMID- 19904411 TI - Simple practical approach for sample loading prior to DNA extraction using a silica monolith in a microfluidic device. AB - A novel DNA loading methodology is presented for performing DNA extraction on a microfluidic system. DNA in a chaotropic salt solution was manually loaded onto a silica monolith orthogonal to the subsequent flow of wash and elution solutions. DNA was successfully extracted from buccal swabs using electro-osmotic pumping (EOP) coupled with in situ reagents contained within a 1.5% agarose gel matrix. The extracted DNA was of sufficient quantity and purity for polymerase chain reaction (PCR) amplification. PMID- 19904412 TI - Towards an electrowetting-based digital microfluidic platform for magnetic immunoassays. AB - We demonstrate ElectroWetting-On-Dielectric (EWOD) transport and SQUID gradiometer detection of magnetic nanoparticles (MNPs) suspended in a 2 microl de ionized water droplet. This proof-of-concept methodology constitutes the first development step towards a highly sensitive magnetic immunoassay platform with SQUID readout and droplet-based sample handling. Magnetic AC-susceptibility measurements were performed on MNPs with a hydrodynamic diameter of 100 nm using a high-Tc dc Superconducting Quantum Interference Device (SQUID) gradiometer as detector. We observed that the signal amplitude per unit volume is 2.5 times higher for a 2 microl sample droplet compared to a 30 microl sample volume. PMID- 19904413 TI - Removal of background signals from fluorescence thermometry measurements in PDMS microchannels using fluorescence lifetime imaging. AB - We report a method for removing unwanted contributions to fluorescence signals from dyes absorbed in polydimethylsiloxane (PDMS) using fluorescence lifetime imaging microscopy (FLIM). By analysing experimental fluorescence decays using a bi-exponential decay model, we are able to discriminate between emission originating from dye molecules in free solution and those absorbed within the PDMS substrate. Simple image processing allows the unwanted background signal to be removed and thus enables a more accurate assessment of temperature. The efficacy of the approach is demonstrated by measuring temperature changes within a droplet-based PCR device. PMID- 19904419 TI - Solar energy conversion. PMID- 19904414 TI - A microfluidic cell trap device for automated measurement of quantal catecholamine release from cells. AB - Neurons and endocrine cells secrete neurotransmitter and hormones in discrete packets in a process called quantal exocytosis. Electrochemical microelectrodes can detect spikes in current resulting from the oxidation of individual quanta of transmitter only if the electrodes are small and directly adjacent to release sites on the cell. Here we report development of a microchip device that uses microfluidic traps to automatically target individual or small groups of cells to small electrochemical electrodes. Microfluidic channels and traps were fabricated by multi-step wet etch of a silicon wafer whereas Pt electrodes were patterned in register with the trap sites. We demonstrate high-resolution amperometric measurement of quantal exocytosis of catecholamines from chromaffin cells on the device. This reusable device is a step towards developing high-throughput lab-on a-chip instruments for recording quantal exocytosis to increase the pace of basic neuroscience research and to enable screening of drugs that target exocytosis. PMID- 19904420 TI - Introducing a dark reaction to photochemistry: photocatalytic hydrogen from [FeFe] hydrogenase active site model complexes. AB - The light-driven splitting of water into its constituting elements gives access to a valuable fuel from an abundant substrate, using sunlight as the only energy source. Synthetic diiron complexes as functional models of the [FeFe] hydrogenase H2ase enzyme active site have moved into the centre of focus as potentially viable catalysts for the reductive side of this process, i.e. the reduction of protons to molecular hydrogen. The active site of the enzyme, as well as its mimics in an artificial system, are required to accumulate two electrons from single electron transfer events and to combine them with two protons to form hydrogen. Whereas in biology this reaction is not coupled to photosynthesis and thus proceeds in the dark, additional aspects need to be considered when designing a functional artificial system for the light-driven reduction of protons. Suitable photosensitizers have to be chosen that not only provide sufficient driving force for the reduction of the synthetic diiron catalyst, but also allow for selective excitation to minimize photodegradation. Electron transfer efficiencies have to be optimized for all steps and the sequential nature of the catalyst reduction requires a sufficient stability of potentially labile intermediates of the catalytic cycle. In this perspective, systems for the light-driven conversion of protons to molecular hydrogen are discussed where the catalyst is based on model complexes of the [FeFe] H2ase active site. Covalently linked dyads, supramolecular assemblies and multi-component systems will be examined with an emphasis on mechanistic electron transfer schemes, the properties of the individual components, their scope and their potential limitations. PMID- 19904421 TI - How algae produce hydrogen--news from the photosynthetic hydrogenase. AB - Green algae are the only known eukaryotes capable of oxygenic photosynthesis which are equipped with a hydrogen metabolism. Hydrogen production is light dependent, since the [FeFe] hydrogenases are coupled to the photosynthetic electron transport chain via ferredoxin. Algal [FeFe] hydrogenases are one of the most active biocatalysts for the evolution of hydrogen. Therefore, special interest exists in the biophysical characterization and biotechnological usage of these [Fe-S] enzymes. This review traces the discovery of this interesting class of proteins. Recent findings allow insight into the electronic structure and configuration of the [FeFe] hydrogenase active site (H-cluster). Emphasis is placed on novel discoveries of the hydrogenase interaction with its natural electron donor ferredoxin and the mechanism of enzyme inactivation through oxygen. PMID- 19904422 TI - Recombinant and in vitro expression systems for hydrogenases: new frontiers in basic and applied studies for biological and synthetic H2 production. AB - This review focuses on recent progress in developing heterologous and recombinant expression as well as in vitro maturation systems for the biosynthesis of active [FeFe] and [NiFe]-hydrogenases, which catalyze the reversible reaction, H2 <--> 2e- + 2H+. Activities of [FeFe] and [NiFe]-hydrogenases produced from different recombinant and in vitro maturation approaches are compared. Examples of how hydrogenase expression supports basic and applied studies of these enzymes are presented, and barriers to achieving more viable biological and synthetic H2 production systems and catalysts are addressed. PMID- 19904423 TI - Solar energy conversion in a photoelectrochemical biofuel cell. AB - A photoelectrochemical biofuel cell has been developed which incorporates aspects of both an enzymatic biofuel cell and a dye-sensitized solar cell. Photon absorption at a porphyrin-sensitized n-type semiconductor electrode gives rise to a charge-separated state. Electrons and holes are shuttled to appropriate cathodic and anodic catalysts, respectively, allowing the production of electricity, or a reduced fuel, via the photochemical oxidation of a biomass derived substrate. The operation of this device is reviewed. The use of alternate anodic redox mediators provides insight regarding loss mechanisms in the device. Design strategies for enhanced performance are discussed. PMID- 19904424 TI - Transcriptional regulation of the cyanobacterial bidirectional Hox-hydrogenase. AB - The overall processes of transcription and its regulation have advanced significantly in the last years, making our understanding of prokaryotic biology more complex and detailed. In fact, a systematic study of different aspects of transcriptional regulation opens up outstanding opportunities to improve and develop the perception of complex reaction mechanisms, genetic processes and cell functions. In close connection to the cyanobacterial bidirectional hydrogenase, the main hydrogen-evolving enzyme in non-nitrogen fixing strains, two novel transcription factors have received increasing attention over the past five years: a LexA-related protein and the AbrB-like family members. Recent work on these regulators has produced new insights and advances towards the understanding (and possible interconnection) of several regulatory networks in cyanobacteria, namely nitrogen metabolism, redox response, toxin production, CO2 concentrating mechanisms and hydrogen metabolism. The fact that a LexA-related protein and AbrB like family members have been co-purified in independent laboratories studying different sets of cyanobacterial genes suggests a possible common and/or complementary function of these regulators. In this review, we summarize the knowledge gained thus far regarding the transcriptional regulation of the cyanobacterial bidirectional hydrogenase, with special focus on the above mentioned transcription factors. Moreover, we discuss several additional points that warrants further investigation to increase our knowledge in this fast evolving research field. PMID- 19904425 TI - Photoinduced water oxidation sensitized by a tetranuclear Ru(II) dendrimer. AB - A multimetallic ruthenium(II) dendrimer is used for the first time to photosensitize dioxygen production from water by IrO2 nanoparticles; the system is more efficient than an analogous system based on the more commonly used [Ru(bpy)3]2+-type photosensitizers, in particular for the ability of the dendrimer to take advantage of the red portion of the solar spectrum. PMID- 19904426 TI - Charge separation and fullerene triplet formation in blend films of polyfluorene polymers with [6,6]-phenyl C61 butyric acid methyl ester. AB - Transient and steady state optical spectroscopies were used to study thin films made from a series of polyfluorene polymers blended with [6,6]-phenyl C61 butyric acid methyl ester (PCBM) in order to determine the influence of polymer ionisation potential on photoinduced charge separation. We find that the energy of the charge separated state DeltaE(CS), given by the energy difference between the ionisation potential of the polymer and the electron affinity of the fullerene, must be smaller than a threshold value of about 1.6 eV for charge separation to occur. When DeltaE(CS) is greater than this threshold, PCBM triplet formation is observed in preference to charge pair generation. If DeltaE(CS) is similar to the threshold value, both PCBM triplet formation and charge separation occur in the blend film, with a tendency for charge separation to dominate over PCBM triplet formation as PCBM concentration increases. The mechanism of triplet formation is believed to be energy transfer to the PCBM singlet state followed by intersystem crossing. The threshold value of DeltaE(CS) is found to be similar to the PCBM singlet energy. PMID- 19904427 TI - Dendrimer light-harvesting: intramolecular electrodynamics and mechanisms. AB - In the development of highly efficient materials for harvesting solar energy, there is an increasing focus on purpose-built dendrimers and allied multi chromophore systems. A proliferation of antenna chromophores is not the only factor determining the sought light-harvesting efficiency; the internal geometry and photophysics of these molecules are also crucially important. In particular, the mechanisms by means of which radiant energy is ultimately trapped depends on an intricate interplay of electronic, structural, energetic and symmetry properties. To better understand these processes a sound theoretical representation of the intramolecular electrodynamics is required. A suitable formalism, based on quantum electrodynamics, readily delivers physical insights into the necessary excitation channelling processes, and it affords a rigorous basis for modelling the intramolecular flow of energy. PMID- 19904428 TI - Passivation of nanocrystalline TiO2 junctions by surface adsorbed phosphinate amphiphiles enhances the photovoltaic performance of dye sensitized solar cells. AB - We report a new class of molecular insulators that electronically passivate the surface of nanocrystalline titania films for high performance dye sensitized solar cells (DSC). Using electrical impedance measurements we demonstrate that co adsorption of dineohexyl bis-(3,3-dimethyl-butyl)-phosphinic acid (DINHOP), along with the amphiphilic ruthenium sensitizer Z907Na increased substantially the power output of the cells mainly due to a retardation of interfacial recombination of photo-generated charge carriers. The use of phosphinates as anchoring groups opens up new avenues for modification of the surface by molecular insulators, sensitizers and other electro-active molecules to realize the desired optoelectronic performance of devices based on oxide junctions. PMID- 19904429 TI - Calculations of interfacial interactions in pyrene-Ipa rod sensitized nanostructured TiO2. AB - Pyrene chromophores carrying different rigid rod spacer groups (ethynylene, ethynylene-phenylene-ethynylene, and ethynylene-bicyclo[2.2.2]octylene ethynylene) and bound to TiO2 nanostructured materials via an isophthalic acid (Ipa) anchor group have been investigated using quantum chemical calculations in order to elucidate structural and electronic properties of dye-sensitized semiconductor structures capable of long-range photoinduced interfacial electron transfer. The calculations are used to study firstly the effect of the anchor and spacer groups on the electronic properties of the pyrene-dyes, secondly the binding of isophthalic acid to nanostructured TiO2, and thirdly the interfacial electronic interactions for dye-sensitized nanostructured TiO2 relevant to dye sensitized solar cell applications. Together, these calculations provide theoretical insights into the effect of incorporating rigid rod anchor-cum-spacer group motifs in sensitizers for e.g. solar cell applications. In particular, the calculations help to rationalize the strong influence of the rods on the photophysical properties of the sensitizers in terms of electronic interactions between the individual chromophore, spacer, and anchor segments, as well as to provide information about interfacial electronic interactions of interest for the capabilities of the rods to act as efficient mediators of photoinduced interfacial charge separation. PMID- 19904430 TI - Structure-property relationships of small bandgap conjugated polymers for solar cells. AB - Conjugated polymers as electron donors in solar cells based on donor/acceptor combinations are of great interest, partly due to the possibility of converting solar light with a low materials budget. Six small bandgap polymers with optical bandgap ranging from 1.0-1.9 eV are presented in this paper. All polymers utilize an electron donor-acceptor-donor (DAD) segment in the polymer backbone, creating a partial charge-transfer, to decrease the bandgap. The design, synthesis and the optical characteristics as well as the solar cell characteristics of the polymers are discussed. The positions of the energy levels of the conjugated polymer relative to the electron acceptor are of significant importance and determine not only the driving force for exciton dissociation but also the maximum open-circuit voltage. This work also focuses on investigating the redox behavior of the described conjugated polymers and electron acceptors using square wave voltammetry. Comparing the electrochemical data gives important information of the structure-property relationships of the polymers. PMID- 19904431 TI - Probing the nanoscale phase separation in binary photovoltaic blends of poly(3 hexylthiophene) and methanofullerene by energy transfer. AB - The generation of charge carriers in organic photovoltaic devices requires exciton diffusion to an interface of electron donor and acceptor materials, where charge separation occurs. We report a time resolved study of fluorescence quenching in films of poly(3-hexylthiophene) containing a range of fractions of the electron acceptor [6,6]-phenyl-C61-butyric acid methyl ester (PCBM). We show that energy transfer from P3HT to PCBM helps to bring excitons to the interface, where they dissociate into charge carriers. Fluorescence quenching in blends with < or = 50 wt% of PCBM is controlled by exciton diffusion in P3HT. This allows us to estimate the average size of PCBM domains to be about 9 nm in the 1:1 blend. The implications for polymer solar cells are discussed. PMID- 19904432 TI - Two tetranuclear Mn-complexes as biomimetic models of the oxygen evolving complex in Photosystem II. A synthesis, characterisation and reactivity study. AB - In this work we report the preparation of two metallamacrocyclic tetranuclear manganese(II) complexes, [L1(4)Mn4](ClO4)4 and [L2(4)Mn4](ClO4)4 where L1 and L2 are the anions of the heptadentate ligands 2-((2-(bis(pyridin-2 ylmethyl)amino)ethyl)(methyl)amino)acetic acid and 2-(benzyl(2-(bis(pyridin-2 ylmethyl)amino)ethyl)amino)acetic acid), respectively. The complexes have been fully characterized by ESI-MS, elemental analysis, single-crystal X-ray diffraction, magnetic susceptibility, and EPR spectroscopy. Electrochemical reactions as well as reactions with different chemical redox reagents have been performed and a reversible two electron oxidation per manganese ion has been identified. The reaction of [L1(4)Mn4](ClO4)4 with oxone or cerium(IV) results in the evolution of oxygen which makes this system interesting for future studies in the search for a functional mimic of the oxygen evolving complex in Photosystem II. PMID- 19904433 TI - Enhancement of photocatalytic activity of zinc-germanium oxynitride solid solution for overall water splitting under visible irradiation. AB - Overall water splitting to form hydrogen and oxygen over a heterogeneous photocatalyst using solar energy is a promising process for clean and renewable hydrogen production on a large-scale. The ZnGeN2-ZnO solid solution photocatalyst shows photocatalytic activity under visible irradiation. This paper deals with results of photocatalytic reactions over the (Zn(1+x)Ge)(N2O(x)) photocatalyst for overall water splitting. The photocatalyst shows high rates for overall water splitting under visible irradiation, with notable deactivation with time. This study focuses on measuring rates for H2 and O2 evolution to elucidate the cause of the deactivation. Our results showed negligible changes in the structure and the composition of the photocatalyst after the photocatalytic reaction, and thus the changes undetectable by the techniques employed should be the cause of deactivation. This paper also discusses two strategies to improve the photocatalytic activity: metal doping in the oxynitride formulation and post calcination after nitridation. Several characterization carried out in this study led us to conclude, at the current point, that the improvement of photocatalytic activity is ascribed to the reduction in the number of defects in the photocatalyst materials. PMID- 19904435 TI - Symmetric band structures and asymmetric ultrafast electron and hole relaxations in silicon and germanium quantum dots: time-domain ab initio simulation. AB - State-of-the-art time domain density functional theory and non-adiabatic (NA) molecular dynamic simulations are used to study phonon-induced relaxation of photoexcited electrons and holes in Ge and Si quantum dots (QDs). The relaxation competes with productive processes and causes energy and voltage losses in QD solar cells. The ab initio calculations show that quantum confinement makes the electron and hole density of states (DOS) more symmetric in Si and Ge QDs compared to bulk. Surprisingly, in spite of the symmetric DOS, the electron and hole relaxations are quite asymmetric: the electrons decay faster than the holes. The asymmetry arises due to stronger NA coupling in the conduction band (CB) than in the valence band (VB). The stronger NA coupling of the electrons compared to the holes is rationalized by the larger contribution of the high-frequency Ge-H and Si-H surface passivating bonds to the CB relative to the VB. Linear relationships between the electron and hole relaxation rates and the CB and VB DOS are found in agreement with Fermi's golden rule. The faster relaxation of the electrons compared to the holes in the Ge and Si QDs is unexpected and is in contrast with the corresponding dynamics in the majority of binary QDs, such as CdSe. It suggests that Auger processes will transfer energy from holes to electrons rather than in the opposite direction as in CdSe, and that a larger fraction of the photoexcitation energy will be transferred to phonons coupled with electrons rather than holes. The difference in the phonon-induced electron and hole decay rates is larger in Ge than Si, indicating that the Auger processes should be particularly important in Ge QDs. The simulations provide direct evidence that the high-frequency ligand modes on the QD surface play a pivotal role in the electron-phonon relaxation dynamics of semiconductor QDs. PMID- 19904434 TI - Water oxidation in photosystem II: oxygen release, proton release and the effect of chloride. AB - Recent reviews on quantum chemical studies of water oxidation in photosystem II have focused on the energetics of O-O bond formation, including the energies of the different S-states. In the present paper, this is extended by discussing the release pathways for dioxygen and protons from the oxygen evolving complex. Based on recent X-ray structures, the effect of adding chloride has also been investigated. Unlike most earlier suggestions, a small, but probably significant, electrostatic effect of adding chloride is found for the higher S-states. For releasing dioxygen in the S4-state, entropy plays a major role. The suggested pathway for proton release in the S1 to S2 transition involves key roles of an outside water and the motion of Asp170. An electron transfer between manganese centra during proton release is also found to be important for a low barrier. PMID- 19904436 TI - Synergistic effect between anatase and rutile TiO2 nanoparticles in dye sensitized solar cells. AB - A synergistic effect between anatase and rutile TiO2 is known, in which the addition of rutile can remarkably enhance the photocatalytic activity of anatase in the degradation of organic contaminants. In this study, mixed-phase TiO2 nanocomposites consisting of anatase and rutile nanoparticles (NPs) were prepared for use as photoanodes in dye-sensitized solar cells (DSSCs) and were characterized by using UV-vis spectroscopy, powder X-ray diffraction and scanning electron microscopy. The addition of 10-15% rutile significantly improved light harvesting and the overall solar conversion efficiency of anatase NPs in DSSCs. The underlying mechanism for the synergistic effect in DSSCs is now explored by using time-resolved terahertz spectroscopy. It is clearly demonstrated that photo excited electrons injected into the rutile NPs can migrate to the conduction band of anatase NPs, enhancing the photocurrent and efficiency. Interfacial electron transfer from rutile to anatase, similar to that in heterogeneous photocatalysis, is proposed to account for the synergistic effect in DSSCs. Our results further suggest that the synergistic effect can be used to explain the beneficial effect of TiCl4 treatment on DSSC efficiency. PMID- 19904437 TI - Beyond the Forster theory of excitation energy transfer: importance of higher order processes in supramolecular antenna systems. AB - Electronic excitation energy transfer in molecular systems is connected with the de-excitation of one molecule and the excitation of the other. Mostly, it can be understood in terms of Forster (or fluorescence) resonance energy transfer. An increasing interest in the optimization of artificial light harvesting systems, however, requires a more detailed study going beyond the standard Forster scheme. There are two main routes to do this. Considering a Coulombic less-strongly coupled system first, coherences among different chromophores may be considered in the framework of perturbation theory with higher-order mechanisms correcting the standard second-order description. Secondly, if inter-chromophore coherences are dominant and delocalized Frenkel-exciton states are formed, it becomes of some importance to study their decay due to the coupling to vibrational degrees of freedom. While we will also comment on this latter mechanism, the first description based on localized excitations will be the main focus. A general higher-order theory resulting in respective transition rates and rate equations is utilized. Its capability is demonstrated when presenting a systematic description of short-range and long-range corrections to the basic Forster mechanism. Accordingly, a unique description of bridge-mediated and photon mediated long-range electronic excitation energy transfer is offered. Moreover, short-range excitation energy transfer appearing as a two electron exchange is also discussed. And, the exciton-exciton annihilation process present at higher optical excitation intensities is described as a direct higher-order transition. The related higher-order vibrational correlation functions are presented and estimated for the reference case where the coupling to vibrational degrees of freedom either of intra-molecular or inter-molecular type is reduced to a simple electronic state dephasing process. PMID- 19904438 TI - Maximizing H2 production in Photosystem I/dithiol molecular wire/platinum nanoparticle bioconjugates. AB - Photosystem I/dithiol molecular wire/nanoparticle bioconjugates have been shown to photocatalytically generate dihydrogen at linear rates when continuously illuminated. In an effort to maximize H2 production, the pH and ionic concentration of the solution, the mobility of the electron donor, the length and degree of saturation of the molecular wire, and the intensity of the light were systematically investigated. Optimal conditions included a solution buffered at pH 6.0, cross-linked plastocyanin, rebuilt spinach PS I, and the use of 1,4 benzenedithiol to connect PS I to the Pt nanoparticle. Illumination of this optimized Photosystem I/dithiol molecular wire/nanoparticle bioconjugate at a light intensity of 70 microE m(-2) s(-1) generated a measured one time H2 production rate of 312 micromol H2 mg Chl(-1) h(-1). PMID- 19904442 TI - Neonatal screening--a global perspective. PMID- 19904439 TI - Binuclear TiOMn charge-transfer chromophore in mesoporous silica. AB - An all-inorganic heterobinuclear chromophore consisting of Ti(IV) oxo-bridged to a Mn(II) center has been assembled on the surface of silica pores of MCM-41 material. The key step of covalent attachment on the pore surface is the reaction of a Mn(II) precursor featuring weakly held CH3CN ligands with the OH group of a previously anchored titanol site. The optical diffuse reflectance spectrum reveals a Ti(IV)OMn(II) --> Ti(III)OMn(III) metal-to-metal charge-transfer (MMCT) absorption extending from the UV throughout to visible into the red spectral region. FT-IR, FT-Raman and optical spectroscopy confirm that the material is free of Mn oxide clusters, while EPR and Mn K-edge X-ray absorption spectra indicate that the donor center is predominantly in oxidation state +2. In situ FT IR spectroscopy allowed detection of visible light-induced redox chemistry of the MMCT unit using O2 (18O2) and methanol as acceptor and donor probe molecules, respectively. Formate and water were observed as primary products, with methyl formate emerging as a secondary condensation product. The observed photochemistry demonstrates that excitation of the Ti(IV)OMn(II) --> Ti(III)OMn(III) results in complete transfer of an electron from donor to acceptor center, with the charge separation sufficiently long lived for initiation of redox chemistry to occur. With donor and acceptor redox potentials appropriate for driving multi-electron catalysts for water oxidation, proton or CO2 reduction, the TiOMn(II) unit is an attractive candidate as a charge-transfer chromophore in a solar fuel generating system. PMID- 19904443 TI - Newborn screening for all identifiable disorders with tandem mass spectrometry is cost effective: supporting arguments. AB - Tandem mass spectrometry (MS/MS) has become increasingly popular as the preferred technology for detecting inborn errors of metabolism in newborn screening (NBS) programmes. Its sensitivity and specificity for detecting numerous metabolic conditions is well-documented. As a NBS technology, there are continuing questions about whether MS/MS should be utilised to the fullest when such usage may mean detecting and reporting analytical findings that could lead to differentiating and diagnosing for which treatment efficacy may not yet be proven. As part of a friendly debate to educate conference attendees on both sides of a somewhat controversial issue, 2 papers awere presented giving information supporting or questioning the cost effectiveness of full scan usage and reporting when using MS/MS in NBS. Reported here are some of the supporting arguments. PMID- 19904444 TI - Newborn screening for all identifiable disorders with tandem mass spectrometry is cost effective: the negative case. AB - Tandem mass spectrometry has become increasingly popular as the preferred technology for detecting inborn errors of metabolism in newborn screening programmes. Its sensitivity and specificity for detecting numerous inborn errors has been well documented. However, there are continuing questions about whether the technology should be used to the fullest when such usage may mean detecting and reporting analytical findings that could lead to diagnosing conditions for which clinical outcome is unclear and treatment may not be needed, or treatment efficacy may not yet be proven and cost effectiveness is unlikely. As part of a friendly debate to educate conference attendees on both sides of somewhat controversial issues, these 2 papers at the conference presented some of the information supporting or questioning the cost effectiveness of full scan usage and reporting in tandem mass spectrometry newborn screening. Reported here are some of the questioning arguments. PMID- 19904445 TI - Issues on universal screening for galactosemia. AB - Galactosemia is an inborn error of galactose metabolism, caused by an abnormality in the conversion of galactose and uridine diphosphoglucose to glucose-1 phosphate and uridine diphosphogalactose through the action of 3 sequential enzymes: galactokinase (GALK), galactose- 1-phosphate uridyltransferase (GALT), and uridine phosphogalactose 4-epimerase (GALE). The advent of newborn screening brought hope with early diagnosis and prompt treatment. Newborn screening advocates have pushed for inclusion of galactosemia in the newborn screening panel. However, reports of complications despite early treatment have questioned the merits of universal screening. This paper presents issues in favour and against universal newborn screening for galactosemia. PMID- 19904446 TI - Neuro-developmental deficits in early-treated congenital hypothyroidism. AB - This paper summarises the current evidence on neuro-developmental deficits in the early (< 1 month of age) treated congenital hypothyroid and the influencing factors. A literature search revealed only few citations that compared outcome with matched controls. In all but one, the median age of treatment onset was >2 weeks. Mean Global IQ scores are about 10 points lower and remain identifiable in adulthood. Verbal and performance scores are usually similar. Deficits persisting into adolescence and adulthood involve the visuomotor, memory, attention and posture domains. Lower academic performance is common in the early years. Prenatal factors associated with a worse prognosis are aetiology (dysgenesis), low birth weight, associated complications and severity of hypothyroidism. Postnatal factors are age at onset of treatment (>1 month), lower thyroxine dose at onset (<8 mcg/kg/day), late normalisation of thyroid function (>2 weeks after treatment), and a lower socio economic family status. The author proposes the evaluation of a multi centre cohort with a median age of treatment onset <1 week, TSH normalisation by <3 weeks with treatment thyroxine levels maintained in the 3rd quartile for age. The outcome of this cohort should indicate if current targets in management need to be revised. PMID- 19904447 TI - Eliminating iodine deficiency: obstacles and their removal. AB - Iodine deficiency remains a global concern for developing countries and some industrialised countries. Iodine deficiency is the most common cause of preventable mental retardation, posing a threat to the social and economic development of countries. Initiatives were developed and instituted to accelerate progress to achieve the goal of universal salt iodisation (USI). However, these efforts were not successful in eliminating iodine deficiency disorders (IDD) in some countries. Every year, 50 million children are born without the protection that iodine offers to the growing brain and body and about 18 million suffer some significant degree of mental impairment. The World Health Organization (WHO), United Nations Children's Fund (UNICEF) and non-governmental organisations assist to ensure that populations at risk have access to iodised salt. This paper will review the highlights of iodine deficiency and present the experiences in the various countries in Asia, i.e. assessments of the situation, action plans, and obstacles to implementation. PMID- 19904448 TI - Outcome of early cochlear implantation. AB - INTRODUCTION: Universal newborn hearing screening facilitates early detection of congenital hearing loss. A child found to have severe to profound hearing loss may require a cochlear implant to access sounds in the speech frequency range. MATERIALS AND METHODS: This retrospective study compared the speech perception outcomes of children implanted at 2 years and below (C1) with those implanted later (C3). Baseline and post-implant speech perception scores were recorded using IT-MAIS, TACL-R or PPVT. The percentage of improvement was calculated for each group and statistical significance was determined using the Student's t test. RESULTS: The median follow-up period for C1 (n = 29) and C3 (n = 29) was 29 months (range, 6 to 29 months) and 20 months (range, 5 to 32 months) respectively, which was not statistically significant. Although both groups recorded post-implant improvement of speech reception scores, the difference in the degrees of improvement was statistically significant (P = 0.034). CONCLUSION: More rapid development of speech perceptive skills was achieved in children who were implanted early. Early implantation therefore, enables children to develop good core listening skills and to potentially develop spoken language at a young age. This enhances successful integration into mainstream pre-schools which provide the environment for the early nurturing of social and cognitive skills. PMID- 19904449 TI - Impact of the national hearing screening programme in China. AB - China has a large population with different levels of medical care among the eastern, central and western areas. The national universal newborn hearing screening (UNHS) programme was initiated in 1999 and then progressively implemented nationwide. A "National UNHS Experts Group" was set up, formulating the national UNHS administration rules and technological specifications. 3 March had been named as national "ear-care day" since 2000 and such social activities help make deafness prevention work more widely accepted. UNHS in China presently has 3 phases due to disparities in economic development. 1) Implementation in stages: in economically under-developed areas. 2) Implementation completed: in the coastal cities. 3) Beyond basic UNHS: i) Development of a completed UNHS system including follow-up and quality control based on the neonatal disease screening system, ii) Exploration of a new public health care programme: simultaneous screening of newborn hearing and ocular disease, iii) Carrying out of a multi-centre study on high-risk factors and GJB2 gene mutations in newborn with non-syndromic hearing impairment. The incidence of newborn bilateral hearing loss was 2.22 per 1000, and 2.74 per 1000 for unilateral hearing loss. Though UNHS have been carried out widely in the eastern parts of China, there are difficulties for its implementation in the western regions. Economic development and technical expertise are the main restricting factors. PMID- 19904450 TI - Early intervention for hearing impairment: appropriate, accessible and affordable. AB - Before the onset of universal newborn hearing screening, children with hearing loss are not identified until they fail to meet important speech and language milestones at 2 years old and beyond. With the current widespread implementation of universal newborn hearing screening programmes, more infants with hearing loss can now be identified in the first few weeks of life and be fitted with amplification within the first few months. This presentation aims to discuss the adverse effects of hearing loss in a child's development. More importantly, it will highlight the value of early identification and early intervention and how these can maximise a child's healthy development of speech, language, academic, emotional and psychosocial skills, thereby facilitating his/her successful integration into mainstream society. In Singapore, universal newborn hearing screening is in place in major hospitals and polyclinics with childbirth services, making it accessible to all families with newborn babies. There are also a number of early intervention programmes that provide rehabilitation services focusing on the development of communication skills of children with hearing impairment. With the availability of services and abundant government support, any child with hearing loss should be identified as soon as possible and provided with early, appropriate intervention. A hearing impairment is said to be an "invisible disability," yet it is the most common major birth defect. In Singapore, one in 1000 babies are born with severe to profound hearing loss and about 5 in 1000 with lesser degrees of hearing loss. Several surveys indicate that between 1 and 3 percent of all children suffer from hearing loss. PMID- 19904451 TI - A multidisciplinary approach to paediatric hearing loss: programme at the centre for hearing intervention and language development, National University Hospital, Singapore. AB - The objective is to describe the multidisciplinary management programme at the National University Hospital (NUH) in Singapore for children with hearing impairment (HI). Over 99.95% of babies born at NUH have hearing tested with both otoacoustic emission and automated auditory brainstem response tests by 6 weeks of age. The referral rate to Otolaryngology is 0.5%. Acquired causes of congenital HI are decreasing. Thirty percent of patients at NUH with idiopathic congenital sensorineural HI have DFNB1/ GJB6 Connexin 26 HI. CT scan or MRI imaging has a higher diagnostic yield when there is unilateral, fluctuating or non-Connexin 26 related HI. Routine electrocardiogram and Opthalmology evaluations will exclude associations of fatal cardiac rhythm anomaly and retinopathy. Other investigations are directed by history and clinical examination. There is now a very wide range of increasingly sophisticated medication, neuro-otologic external, middle and inner ear surgery, hearing aids, middle ear implants and cochlear implants available to improve hearing. A multidisciplinary team from neonatology, paediatrics, otolaryngology, audiology, auditory verbal and speech therapy, ophthalmology, radiology, and psychology working closely with the child, family and schools is needed to develop a cost effective and comprehensive management programme for paediatric HI. PMID- 19904452 TI - Current understanding of auditory neuropathy. AB - Auditory neuropathy is defined by the presence of normal evoked otoacoustic emissions (OAE) and absent or abnormal auditory brainstem responses (ABR). The sites of lesion could be at the cochlear inner hair cells, spiral ganglion cells of the cochlea, synapse between the inner hair cells and auditory nerve, or the auditory nerve itself. Genetic, infectious or neonatal/perinatal insults are the 3 most commonly identified underlying causes. Children usually present with delay in speech and language development while adult patients present with hearing loss and disproportionately poor speech discrimination for the degree of hearing loss. Although cochlear implant is the treatment of choice, current evidence show that it benefits only those patients with endocochlear lesions, but not those with cochlear nerve deficiency or central nervous system disorders. As auditory neuropathy is a disorder with potential long-term impact on a child's development, early hearing screen using both OAE and ABR should be carried out on all newborns and infants to allow early detection and intervention. PMID- 19904453 TI - Improving child health--newborn screening for all? AB - Over the last 40 years newborn screening has been an undoubted success and many thousands of children have been saved from mental retardation and other problems because of early diagnosis of their disorders. Now many diseases can be diagnosed early by newborn screening and many more are on the horizon. It must be a long term goal to extend newborn screening tests to all children but, in areas of the world where healthcare delivery is insufficient, solving other health problems has to take precedence over introducing newborn screening. If it is decided to introduce newborn screening in a region where currently there is none screening for congenital hypothyroidism alone should be started before anything else at all is attempted so that proper systems can be put in place. There is an exciting future for newborn screening ahead. If new programmes are approached with proper caution maximal benefit will be achieved from newborn screening, which is one of the few clearly effective preventive strategies in healthcare. PMID- 19904454 TI - Establishing a universal newborn hearing screening programme. AB - As congenital hearing impairment has a worldwide incidence of 4 to 5 per 1000 babies and is thus one of the most common congenital problems seen today, universal newborn screening has a crucial role to play in its early detection and intervention. It provides the opportunity for better outcomes and normal language development. Prior to embarking on a screening programme, the newborn population and the current health care system should be analysed to select the best method of coverage. The screening tool and protocol, communication of results, as well as the follow-up measures should be clearly determined and tested. The multidisciplinary team required should be provided with the necessary information. Parents need to be educated about the importance of early hearing screening. Data management and surveillance should be established in a systematic manner. The costs of the programme should be carefully anticipated and funding sources determined. Finally, support for the programme should be sought from governmental or public health bodies, to ensure the success of the programme. Legislation can be considered if necessary. PMID- 19904455 TI - Spectrum of inherited metabolic disorders in Malaysia. AB - Issues pertaining to the diagnosis and management of inborn errors of metabolism (IEM) in Malaysia included low awareness of atypical and variable presentations in IEMs leading to delayed diagnosis or treatment, absence of reliable population data on IEMs and involvement of multiple siblings in the same family due to consanguinity. The importance of careful family history taking and genetic counselling are emphasised. Selected testing of ill infants and children for IEM yielded a positive 2% (264/13,500) results for IEMs in Malaysia. Out of the 264 patients, the spectrum of IEMs in Malaysia included organic acidurias (98), aminoacidopathies (78), urea cycle defects (54), neurotransmitter conditions (12) and lysosomal disorders, mainly mucopolysaccharidosis (14). Confirmatory studies of IEMs are an important aspect of management of IEMs. There is a need for more metabolic specialists and funding for diagnosis and treatment of IEMs in Malaysia. Long-term care issues and cost-effectiveness of IEM therapy, supportive and preventive aspects will need further studies in Malaysia. PMID- 19904456 TI - Disorders of the carnitine cycle and detection by newborn screening. AB - Carnitine is necessary for transport of long-chain fatty acids into mitochondria, to enter the beta-oxidation cycle. Four carnitine cycle defects have been described. The carnitine transporter mediates carnitine transport across the plasma membrane. Symptoms include hypoketotic hypoglycaemia and cardiomyopathy. Some affected subjects are asymptomatic. Newborn screening detects very low levels of free carnitine in some but not all. Carnitine palmitoyltransferase type IA (CPTI) transports long-chain fatty acyl-CoAs across the outer mitochondrial membrane. Affected infants have hypoketotic hypoglycaemia with catabolic stress, but otherwise remain well. Newborn screening tests reveal elevated free carnitine, (elevated C0/C16+C18). Sensitivity is unclear and confirmation needs leukocyte or fibroblast assays. Carnitine-acylcarnitine translocase transfers fatty acylcarnitines across the inner mitochondrial membrane. The most common presentation is sudden death in the first days. Carnitine palmitoyltransferase type II (CPTII) converts long-chain acylcarnitines to long-chain acylCoAs for beta-oxidation. Severe deficiency is lethal. Newborn screening for both disorders reveals elevated palmitoylcarnitine and enzymology or mutation analysis is needed for diagnosis. Late-onset CPTII is the most common disorder, presenting as muscle pain and rhabdomyolysis on severe exercise. All 4 disorders can be detected by newborn screening, with variable sensitivity. Late-onset CPTII probably cannot be detected. Carnitine transporter, CPTI and late-onset CPTII have proven treatment strategies. PMID- 19904457 TI - Fatty Acid oxidation defects. AB - Fatty acid oxidation defects (FAODs) are a group of inborn errors of metabolism that carry a risk for morbidity and mortality. Successful management of these conditions involves early diagnosis, proper management of clinical situations that predisposes to metabolic crisis as well as adequate treatment of a metabolic crisis. The advent of tandem mass spectrometry based newborn screening has reduced the morbidity and mortality in some FAODs. This article discusses how to recognise FAODs and how to manage them. PMID- 19904458 TI - Diagnosis of tetrahydrobiopterin (BH4) responsive mild phenylketonuria in Japan over the past 10 years. AB - BACKGROUND: A novel therapeutic strategy for phenylketonuria (PKU) has been initiated in Japan. Hyperphenylalaninemia (HPA) results from a phenylalanine hydroxylase (PAH) enzyme deficiency or a deficiency of its cofactor, tetrahydrobiopterin (BH4). BH4 can normalize blood phenylalanine levels in BH4 deficiency, but typically not in PKU. However, since 1999 it has been reported that many HPA patients (serum phenylalanine <20 mg/dL) showed a gradual decrease of serum phenylalanine levels after 24 hours from BH4 loading. The BH4 responsiveness seems to be regulated in mild PKU by PAH mutations, and affected by the BH4 dose and administration period. METHODS AND RESULTS: In 2002 we formulated a provisional diagnostic criteria for patients with BH4-responsive PAH deficiency, and newly diagnosed 19 patients in 100 HPA cases between 2002 and 2006. The incidence in the recent 5 years for BH4-responsive mild PKU among patients with PAH deficiency was 25 %. CONCLUSION: A total of 31 patients was detected in the past 10 years, and the incidence detected using the provisional diagnostic criteria had increased to 25% among PAH deficient patients. BH4 treatment for BH4-responsive mild PKU is a new and effective pharmacotherapy, which replaces or liberalises the phenylalanine-restricted diets for a considerable number of mild PKU patients. PMID- 19904459 TI - Disorders of vitamin B12 metabolism presenting through newborn screening. AB - Elevated propionyl C3 carnitine is the most common abnormality seen in tandem mass spectrometry newborn screening profiles, with an incidence of 0.15% seen in our South Australian newborn screening programme. The most common cause for this result in our population is vitamin B12 deficiency but differential diagnoses include the inherited disorders of propionic and methylmalonic acid metabolism and cobalamin deficiencies. An approach to confirmatory testing and subsequent management of infants with elevated propionic carnitine is presented. PMID- 19904460 TI - Glucose-6-phosphate dehydrogenase deficiency: correlation between the genotype, biochemistry and phenotype. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common genetic enzyme defect present in many people from African, Middle Eastern, Mediterranean and Asian countries. Individuals with the enzyme deficiency may remain asymptomatic, develop an acute haemolytic crises to infections or Fava beans, neonatal jaundice or chronic non-spherocytic haemolytic anaemia. Electrophoretic mobility may be fast, slow or normal. Over 160 mutations have been described, mostly due to single amino acid substitution. Although correlation of the genotype and biochemistry with the clinical phenotype of G6PD deficient individuals remains somewhat variable, there is better correlation among individuals presenting with chronic non-spherocytic haemolytic anaemia, which is related to the NADP structure of the enzyme. PMID- 19904461 TI - External quality assurance programme for newborn screening of glucose-6-phosphate dehydrogenase deficiency. AB - INTRODUCTION: The nationwide neonatal screening for glucose-6-phosphate dehydrogenase (G6PD) deficiency in Taiwan was started on 1 July 1987. A network of G6PD referral hospitals distributed all around Taiwan was organised for follow up, confirmatory testing, medical care and genetic counselling. To assess the reliability of confirmatory and screening tests, an external quality assurance (QA) programme for G6PD assay was developed. MATERIALS AND METHODS: Lyophilised quality control (QC) materials and dried blood spots were prepared from erythrocytes and whole blood for confirmatory and screening tests, respectively. The external QA surveys were carried out every 1 to 2 months. The QA results were evaluated and compared to the consensus result and reference value. The test results were submitted through internet by participating laboratories and the summary reports were published on a webpage (http:// www.g6pd.tw) within 2 weeks. RESULTS: Twenty-one referral laboratories in Taiwan and 16 screening laboratories in Germany, Lebanon, Mainland China, Philippines, Thailand, Taiwan, Turkey, and Vietnam have been participating in the QA programme. From 1988 to 2007, 144 QA surveys for confirmatory testing were sent to referral laboratories. Among the 2,622 reports received, 292 (11.1%) were found to be abnormal. Interlaboratory coefficient of variation (CV) for the confirmatory test has reached below 10% in recent years. The significant improvement in interlaboratory CV was found to be correlated with the preventive site visits to the referral laboratories since November 2004. From 1999 to 2007, 52 external QA surveys for the screening test were performed. Among 504 reports received, 97 (19.2%) were found to be abnormal. From the 5040 blood spots tested by the screening laboratories, 95 false negative (1.9%) and 187 false positive (3.7%) results were reported. CONCLUSIONS: The external QA programme has been useful for monitoring the performance of the referral hospitals and screening laboratories and helpful for the participating laboratories to improve their test quality. PMID- 19904462 TI - Neuroblastoma screening in Japan: population-based cohort study and future aspects of screening. AB - INTRODUCTION: It is unknown whether screening for neuroblastoma has the benefit of reducing the incidence of advanced diseaseor mortality due to neuroblastoma.Japanese nationwide massscreening for 6 month old infants was launched in 1985 and was performed using quantitative high-performance liquid chromatography (HPLC) between years 1990 to 2003. MATERIALS AND METHODS: We compared the incidence rates (IR) and the mortality rates (MR) per 100,000 births of neuroblastomas diagnosed before 6 years of age between 2 cohorts: children born during the years 1980 to1984 (Pre-screen cohort, n = 7,620,203) and 1990 to1998 (Screen cohort, n = 10,878,918). We then proposed the optimal timing and procedures for future screening. RESULTS: Cumulative IR in the Screen cohortwas significantly higher than the Pre-screen cohort (29.80 vs. 11.96, P <0.0001). On the other hand, IR of neuroblastoma diagnosed after 24 months old in the Screen cohort was significantly lower than in the Pre-screen cohort (P <0.0001). The cumulative MR of the Pre-screen cohort was 5.35, whereas that of the Screen cohort was 2.82 (P <0.0001). CONCLUSIONS: HPLC mass-screening for neuroblastoma at 6 months of age found a marked increase in incidence in younger children (less than 12 month old) and a significant decrease in mortality rates overall. To reduce overdiagnosis of regressing cases and to identify preclinical stages of unfavourable cases, we propose using HPLC-screening at 18 months of age. PMID- 19904463 TI - Legal issues in neonatal screening. AB - Legal issues arise if some persons or institutions feel wrongfully treated whether or not this feeling is justified. In neonatal screening, the following topics may be causing legal issues: no screening programme where such a programme should be (UN Convention for the right of the child); neonate(s) not screened for conditions within the established programme; no consent when it should have been given; error(s) in sampling, analysis, reporting; no follow-up available, error(s) in confirmatory diagnostics and treatment; irregular storage of dried blood spot specimen. Legal issues can be solved easily when responsibilities of parties concerned have been established and documented. Unfortunately, legal systems vary from country to country and what has become "normal" practice in one jurisdiction may still be battled about in another. The management of a neonatal screening programme should try to define as best as possible the performance criteria and to have the programme assessed and accredited to certain internationally accepted standards. It diminishes the chances for errors and it helps to avoid legal issues. PMID- 19904464 TI - Towards universal newborn screening in developing countries: obstacles and the way forward. AB - Newborn Screening is a well recognised public health programme aimed at the early identification of infants who are affected by certain genetic/metabolic/infectious conditions. Early identification of these conditions is particularly crucial, since timely intervention can lead to a significant reduced morbidity, mortality, and associated disabilities in affected infants. Establishing sustainable newborn screening programmes in developing countries poses major challenges as it competes with other health priorities--infectious disease control, immunisation, malnutrition, etc. Despite this, it is imperative that developing countries recognise the importance of newborn screening based on experiences on both developed and developing countries in saving thousands of babies from mental retardation, death and other complications. Some of the critical factors necessary for a successful national newborn screening programme are inclusion of newborn screening among government priorities, funding (including the possibility of newborn screening fees), public acceptance, health practitioners cooperation, and government participation in institutionalising the newborn screening system. This paper presents a historical review of 4 eras of newborn screening in the Asia Pacific, discusses enabling factors leading to successful newborn screening programme implementation, and identifies obstacles that threaten the programme implementation in developing countries. PMID- 19904465 TI - Inborn errors of metabolism presenting as neonatal encephalopathy: practical tips for clinicians. AB - Inborn errors of metabolism constitute an important cause of neurological disease in the neonatal period and can present clinically as encephalopathy. Although it is relatively rare, it is important to have a high index of suspicion. Appropriate investigations and a step-wise approach to diagnosis allow for early institution of treatment and can prevent significant morbidity and mortality. The aim of this article is to give a brief outline of the various inborn errors of metabolism to consider in neonatal encephalopathy and to provide a framework for investigation and diagnosis. PMID- 19904466 TI - Financing newborn screening systems: US experience. AB - Newborn screening (NBS) in the United States (US) has existed since the early 1960s and is required in all 51 state jurisdictions. It is generally recognised that NBS provides a significant public health benefit by preventing or markedly decreasing the adverse medical consequences of conditions included in the screening panel. There is currently no US national NBS policy, so instead there are 51 independent state programmes that vary widely in their policies, infrastructures, procedures and services. Not surprisingly, US NBS programme costs and methods of financing also vary. Surveys have increasingly found a reliance on fees to pay for screening tests, short-term follow-up and other parts of state NBS systems. This article reviews some of the current US NBS financing issues and methodologies. PMID- 19904467 TI - Developmental dysplasia of the hip: universal or selective ultrasound screening? AB - Developmental dysplasia of the hip (DDH) is an intriguing condition that evolves during infancy. It would be thus foolhardy to expect a screening tool at birth to be both highly sensitive and specific. Uncertainty regarding an optimal screening method is compounded by a general lack of sound epidemiological data. Clinical screening remains widely used. Some reports estimated that it did not pick up 60% of children who eventually needed surgery. Ultrasonography, it was hoped, would improve detection rates. There are 2 approaches to ultrasound; universal screening, which is adopted by some European countries, or selective screening of high risk infants. The problems with universal ultrasound screening are high false positive rates and high costs. The benefit was a possible 6- to 10-fold reduction in surgery for late DDH. Similar reductions though had also been reported if ultrasound was used selectively for infants with clinical and historical risk factors. A literature review on this topic is presented. There are pros and cons for both screening strategies. This is reflected in the different protocols that exist among various countries. For healthcare systems that are considering their options, universal ultrasound screening is generally not cost-effective and should not be the preferred screening strategy. PMID- 19904468 TI - Minimising harm from newborn screening programmes. AB - The challenge of newborn screening programmes is to maximise benefits and minimise harms. These harms include pain inflicted as a result of taking the test, reduced by pain relief and training of specimen takers; from false positive and negative test results (impacting both affected families and healthcare professionals), minimised effectively by taking the sample at the correct time, precise and specific tests, appropriate disorder definition, well chosen cut-offs (which may be informed by a large series of diagnosed cases of the screened disorders) second-tier tests, age adjusted normal ranges and anxiety which may be appropriate but limited by the availability of information. Programme audit is important in early detection of problems. PMID- 19904469 TI - Newborn screening in China: phenylketonuria, congenital hypothyroidism and expanded screening. AB - This study was to investigate the current status of neonatal screening in China, to further clarify the incidences of hyperphenylalaninemia (HPA) and congenital hypothyroidism (CH). From 2000 to 2007, a total of 17,961,826 newborns had been screened for HPA and 1527 cases were detected, giving a HPA prevalence of 1:11,763. At the same time, 18,284,745 newborns had also been tested for CH, with 8918 cases being detected (1:2050). It is remarkable that the mean number of newborns screened per year had increased 5 times between 2000 and 2007. In Shanghai, 116,000 newborns were screened using tandem mass spectrometry and 6 different were detected. The overall prevalence of an inborn errors of metabolism identified was 1 in 5800 healthy newborns, with hyperphenylalaninemia being the most common. Neonatal screening had developed rapidly in China in recent years, and a pilot study using tandem mass spectrometry has been started. The biggest challenge is still to increase coverage to the entire country, especially in the mid-western area. PMID- 19904470 TI - Newborn screening in Bangladesh. AB - Newborn screening started in Bangladesh in 1999. The programme started as part of a regional project of the International Atomic Energy Agency (IAEA) to screen for congenital hypothyroidism (CH). In the beginning the IAEA helped the country with equipment, filter papers, reagents, training and expert services. Since 1999, 2 pilot projects to screen newborns for CH were completed. Under these projects some 30,000 newborns were screened and 16 were identified with hypothyroidism. The government of Bangladesh approved a national project in July 2006 to screen newborns in some selected areas of the country for CH. Under the project some 200,000 newborns will be screened and laboratory facilities for newborn screening will be increased. Bangladesh has a large population of about 140 million. With the current birth rate some 2 million new births take place every year. The socio economic situation of the country is also different. Per capita income of the country is one of the lowest in the world. About 85% of babies are still delivered at home. As such newborn screening is a big challenge for Bangladesh. However, the country is trying to overcome these challenges. PMID- 19904471 TI - Newborn screening in Pakistan - lessons from a hospital-based congenital hypothyroidism screening programme. AB - We are living in a time of unprecedented increase in knowledge and rapidly changing technology. Such biotechnology especially when it involves human subjects raises complex ethical, legal, social and religious issues. The establishment of newborn screening programmes in developing countries poses major challenges as it competes with other health priorities like control of infectious diseases, malnutrition and immunization programmes. Despite this, it is imperative that the importance of newborn screening programmes is recognised by developing countries as it has been proven through decades of experience that it saves thousands of babies from mental retardation, death and other serious complications. Pakistan has an estimated population of 167 million inhabitants, 38.3% of whom are under 15 years of age. Pakistan lacks a national programme for newborn screening. However, as individual practice at the local level, Aga Khan University Hospital (AKUH) and a few other hospitals are doing newborn screening for congenital hypothyroidism. The main hurdle in the implementation of newborn screening in Pakistan is the lack of good infrastructure for health. Eighty percent of deliveries take place at home. Moreover, little resources are available for children identified with a genetic condition due to the non existence of genetic and metabolic services in Pakistan. In a 20-year audit of congenital hypothyroid screening at AKUH we found 10 babies with congenital hypothyroidism. However due to missing data links spanning several years, we were unable to calculate its true incidence during this period. In order to estimate the incidence of congenital hypothyroidism (CH) we reviewed in detail data over 10 months in 2008, a period where there was better compliance for repeat thyroid stimulating hormone (TSH) testing, and found 2 babies with CH. This gave an estimated incidence of 1 in 1600 live births. PMID- 19904472 TI - Organising services for IMD in Thailand: twenty years experience. AB - The study of inherited metabolic disorders (IMD) in Thailand is in its infancy when compare with developed countries. Prior to 1987, majority of these disorders were clinically diagnosed since there were only a handful of clinicians and scientists with expertise in inborn errors of metabolism, lack of well-equipped laboratory facilities and government support. In developing countries, inherited metabolic disorders are not considered a priority due to the prevalence of infectious diseases such as HIV infection and congenital infections. A multicentre survey conducted in 1994 and 2001 revealed the existence of numerous cases of IMD from all over the country. Case reports and publications on IMD in Thai (and international) medical journals in past 20 years had undoubtedly raised its awareness among Thai paediatricians and scientists. In 2001, the Genetic Metabolic Centre was first established in Siriraj Hospital Faculty of Medicine, Thailand. Numerous new cases of IMD had been identified since then. PMID- 19904473 TI - Outcome of organic acidurias in China. AB - From June 1998 to May 2007, 9566 urine samples were collected from patients with psychomotor deficits, seizures, vomiting and unconsciousness in Peking University First Hospital. Their urine organic acids profiles were analysed using gas chromatography - mass spectrometry (GCMS), GCMS solution and Inborn Errors of Metabolism Screening System software. In all patients, blood acylcarnitines were analysed using tandem mass spectrometry. One hundred and sixty-eight patients (1.76%) with organic acidurias were detected. Among them, 116 (116/ 168, 69.0%) had methylmalonic aciduria, 63 (54.3%) of these 116 patients had methylmalonic aciduria combined with homocysteinemia. Sixteen (9.5%) of those patients detected with organic acidurias had propionic aciduria, and 15 (8.9%) had multiple carboxylase deficiency. Seven (4.2%) had glutaric aciduria type 1. After dietary treatment, medicine and rehabilitation, clinical improvements were observed in more than half of the patients. Twenty-eight of the 168 patients (16.7%) recovered and led a normal life. The method of urine organic acid analysis by gas chromatography - mass spectrometry and blood acylcarnitines analysis by tandem mass spectrometry have been established and applied successfully in China, namely Beijing, Shanghai, Wuhan and Guangzhou. The prognoses of Chinese patients with organic acidurias have also improved significantly. PMID- 19904474 TI - Improved health and development of children who are deaf and hard of hearing following early intervention. AB - Infants begin to learn language in the earliest months of life. In the absence of early identification and intervention, infants who are deaf or hard-of-hearing experience significant and lasting deficits in language learning, academic achievement, social-emotional development, and quality of life. Evidence is mounting that early identification of infants who are deaf or hard of hearing through newborn hearing screening and intervention by the age of 6 months improves developmental outcomes for these children, especially in the area of language proficiency. Newborn screening programmes, including newborn hearing screening, are typically public health activities aimed at the early identification of infants who are affected by certain congenital disorders: including genetic, metabolic, haematologic, and infectious diseases. Early identification of these conditions is critical, as timely intervention can lead to a significant reduction of morbidity, mortality and associated disabilities in affected infants. For infants with hearing loss, the goal of early identification is to provide early intervention leading to language development and academic achievement commensurate with cognitive ability, and ultimately an improved quality of life. For newborn hearing screening, the definition of early intervention is initiation of intervention by the age of 6 months. Initiatives for newborn hearing screening have spread to every continent and many countries now have well-developed, comprehensive programmes of screening, diagnosis, and early intervention for childhood hearing loss. Although no exact statistics currently exist, tens of millions of infants probably receive newborn hearing screening each year. Because the developmental effects of early intervention for hearing loss are improved and sustained language development, large-scale studies documenting the language outcomes in early-identified children take decades to collect. Furthermore, because full-scale implementation of universal newborn hearing screening has occurred only within the last 5 years in most countries, many early-identified children are still at preschool-age. Thus, documenting these youngsters' language development is a work in progress. Despite these limitations, evidence is mounting that early intervention for childhood hearing loss improves the developmental outcome of these children. PMID- 19904475 TI - Newborn screening in Japan: restructuring for the new era. AB - Nationwide neonatal mass screening for inherited metabolic diseases has started in Japan since 1977. At least 8000 children have probably been spared from handicaps resulting from such diseases over the past 30 years. Recently remarkable changes have been made to the evolving neonatal screening system. Declining birth rate and economic problems in Japan have demanded a more effective neonatal screening system. Development of new innovative screening methods and treatment tools, e.g. tandem mass spectrometry (MS/MS) technology and enzyme replacement therapy for mucopolysaccharidosis (MPS), have facilitated expansion of target diseases in neonatal screening. We have carried out pilot screening using MS/MS in 6 laboratories in Japan. The incidence of inherited metabolic diseases was found to be 1 in 9330 (65 cases out of 606,380 babies screened) during the period between 1997 and 2007. The incidence was lower than those of Europe or USA (about 1 in 4000 to 5000). The disease frequency between unscreened symptomatic cases and asymptomatic cases detected through MS/MS screening were also found to be different. In MS/MS screening, the most common organic acidemia was propionic acidemia, whereas in symptomatic cases, methylmalonic acidemia was the most common. Further study of ethnic diversity in severity of propionic academia is required. The outcomes of patients detected in the MS/MS screening were significantly favourable. The results showed the benefits of MS/ MS screening. The diagnostic support network for gas chromatography-mass spectrometry (GC/ MS) analysis and enzyme determination has also been developed. We have developed an automated system of GC/MS data processing and auto-diagnosis which allowed the GC/MS data processing to be extremely fast and simple. Enzyme evaluation for diagnostic support for screening, including a method using peripheral blood and high performance liquid chromatography (HPLC), and another method of in-vitro probe assay using cultured cells and MS/MS. Furthermore, re-location of screening laboratories for a more efficient screening network will be required such that at least 30,000 samples can be analysed in each laboratory. PMID- 19904476 TI - Clinical applications of molecular genetics: the model of congenital adrenal hyperplasia. AB - Spectacular advances in molecular genetics have enabled the molecular characterisation of many genetic disorders. The clinical applications include: (i) identification of pre-symptomatic and symptomatic affected individuals (monogenic diseases), allowing for early treatment and prevention of complications, (ii) carrier testing for genetic counselling, (iii) pharmacogenetic testing to guide medical treatment, and (iv) susceptibility testing (in polygenic diseases) to determine the risk of developing future disease. Using the model of congenital adrenal hyperplasia (CAH), direct mutational analysis can be applied to: (i) confirm the diagnosis when hormone assays have been equivocal, which would allow for early treatment and prevention of adrenal crisis, (ii) prenatal diagnosis and prenatal treatment in affected females to prevent or reduce prenatal virilisation, (iii) heterozygote carrier identification for genetic counselling, (iv) novel therapeutic applications to optimise treatment, including adjusting the steroid dose based on consistent genotype-phenotype correlations, so as to reduce the incidence of growth inhibiting effects of steroid excess. However, molecular analysis can occasionally be complicated by multiple mutations on one allele, which may potentially affect genotype-phenotype correlations. Hence, molecular genetic analysis of CAH may eventually be adopted as a second tier confirmation of the disease, but is unlikely to replace the current first tier screening assays of precursor steroid metabolites proximal to the enzyme deficiency. PMID- 19904477 TI - Considerations in choosing screening conditions: one (US) approach. AB - The lack of a national policy on newborn screening (NBS) in the United States has resulted in 51 state-specific NBS policies (including 50 states and the District of Columbia). In 2000, a working group of the American Academy of Pediatrics provided a national NBS blueprint for the future. Using this guidance, the Health Resources and Services Administration contracted with the American College of Medical Genetics to: (i) develop a decision-making algorithm for states to use in selecting conditions for screening panels, and (ii) recommend a panel of tests to guide states in their screening requirements. This report outlines and summarises the processes and outcomes leading to the current NBS recommendations in the United States. PMID- 19904478 TI - Diagnosis and management support for an expanded newborn screening programme. AB - The introduction of tandem mass spectrometry technology expands newborn screening and permits early diagnosis of inborn errors of metabolism. Through measurement of a number of acyl carnitines, amino acids and associated ratios, infants at risk of inborn errors of metabolism can be detected. However the increasing availability of the technology places new challenges to areas with established programmes, as well as those without existing newborn screening programmes. Once the technical aspects of tandem mass spectrometry operation are overcome, the initial challenge lies in determination of whether a borderline result is abnormal. Participation in quality assurance and international collaborative programmes is critical to optimise sensitivity and specificity. Some conditions are readily detected, others are more problematic. All positive results must be confirmed with formal testing but the tests required will vary with the disorder. Even after confirmed diagnosis, the significance of the diagnosis for that child may not be clear, as mild forms of disorders, previously thought to be rare, are being recognised by newborn screening programmes. Parents should be provided with easy to understand written information and a management plan. Education of health professionals who may not be familiar with these conditions raises another challenge. Treatment should be supervised by an expert centre and outcome data must be collected to determine the effectiveness of the screening programme in each area. PMID- 19904479 TI - My early experiences in establishing neonatal screening and the reason for regional meetings of the international society for neonatal screening. AB - When I started the neonatal screening in Japan, I could not obtain the necessary information for establishing the national screening system in my own country. Thus, around 1970, I visited H. Bickel, R. Guthrie and several other experts in the field of the neonatal screening in USA and Europe. Through their help, I could learn: (i) the philosophy of the world of screening, (ii) the way to improve the basic techniques in this field and (iii) the way to improve the level of screening. On the other hand, I realised that in some countries, people received imprecise information from non-authoritative sources. I also realised that it was difficult for people in developing countries to meet experts of other countries. Therefore, when I was appointed the first president of International Society of Neonatal Screening (ISNS), I proposed to have the regional meetings held in many areas. In this report, I explained how we were asked to establish the national screening system in Japan through the support of experts around the world. I hope that people will understand the reason why I proposed that regional meetings of the ISNS be held in various locations. PMID- 19904480 TI - Bone mineral density, body composition, and food intake of adolescent runway models. AB - OBJECTIVE: To evaluate the bone mineral density (BMD) and to relate it to the food intake and body composition of adolescent runway models. METHODS: Cross sectional study evaluating 33 models and 33 non-models aged from 15 to 18 years, paired by age and body mass index (BMI). BMD of spine (L1-L4) was evaluated using the dual-energy X-ray absorptiometry technique (Lunar DPX Alpha), and body composition was assessed by means of plethysmography. Food intake was evaluated by a 3-day-food record. RESULTS: The subjects' mean age was 16.75+/-1.04 years, and 24% had BMI below ideal value for their age. BMD values (g/cm2) were similar between models (1.108+/-0.080) and non-models (1.096+/-0.102) (p > 0.05), and 6% of the participants had low BMD for age. We found that the mean energy intake was lower among models as compared to non-models (1,480.93+/-582.95 vs. 1,973.00+/ 557.63 kcal) (p < 0.05) and that most of the adolescents in both groups presented an inadequate consumption of micronutrients, with emphasis to the low calcium intakes. There was only significant correlation between BMD and lean body mass (kg) (r = 0.362 for models and r = 0.618 for non-models) (p < 0.05). CONCLUSION: Although no association was found between BMD, BMI, and intake of nutrients which are important for the bone mineralization process, inadequacies of food intake have an adverse influence on the acquisition of bone mass, which is more effective at this stage of life. PMID- 19904481 TI - Structural traces of past experience in the cerebral cortex. AB - It is widely assumed that changes in the connections between neurons mediate the integration and storage of information in the brain and thereby underlie our ability to learn and remember. In particular, long-term memory is thought to rely on a structural reorganisation of neuronal circuits, but the proof for such a mechanism in the complex mammalian brain remains elusive. Recent advances in scientists' ability to follow structural dynamics of neuronal networks in the intact brain in vivo by means of 2-photon laser scanning microscopy has provided new insight into how information about new experiences might be stored in brain circuits. PMID- 19904482 TI - Evolution of the 2'-5'-oligoadenylate synthetase family in eukaryotes and bacteria. AB - The 2'-5'-oligoadenylate synthetase (OAS) belongs to a nucleotidyl transferase family that includes poly(A) polymerases and CCA-adding enzymes. In mammals and birds, the OAS functions in the interferon system but it is also present in an active form in sponges, which are devoid of the interferon system. In view of these observations, we have pursued the idea that OAS genes could be present in other metazoans and in unicellular organisms as well. We have identified a number of OAS1 genes in annelids, mollusks, a cnidarian, chordates, and unicellular eukaryotes and also found a family of proteins in bacteria that contains the five OAS-specific motifs. This indicates a specific relationship to OAS. The wide distribution of the OAS genes has made it possible to suggest how the OAS1 gene could have evolved from a common ancestor to choanoflagellates and metazoans. Furthermore, we suggest that the OASL may have evolved from an ancestor of cartilaginous fishes, and that the OAS2 and the OAS3 genes evolved from a mammalian ancestor. OAS proteins function in the interferon system in mammals. This system is only found in jawed vertebrates. We therefore suggest that the original function of OAS may differ from its function in the interferon system, and that this original function of OAS is preserved even in OAS genes that code for proteins, which do not have 2'-5'-oligoadenylate synthetase activity. PMID- 19904483 TI - Talua SINE biology in the genome of the Reticulitermes subterranean termites (Isoptera, Rhinotermitidae). AB - Studies on transposable elements in termites are of interest because their genome is in a permanent condition of inbreeding. In this situation, an increase in transposon copy number should be mainly due to a Muller's ratchet effect, with selection against deleterious insertions playing a major role. Short INterspersed Elements (SINEs) are non-autonomous retrotransposons, known to be stable components of eukaryotic genomes. The SINE Talua, first isolated from Reticulitermes lucifugus (Rhinotermitidae), is the only mobile element described so far in termites. In the present survey, Talua has been found widespread in the Isoptera order. In comparison with other non-termite SINEs, Talua diversity and distribution in the Reticulitermes genome demonstrate that Talua is an ancient component of termite genome and that it is significantly associated with other repeats. In particular, the element is found to be involved with microsatellite motifs either as their generator or because inserted in their nearby. Further, two new SINEs and a putative retrotranscriptase-like sequence were found linked to Talua. Talua's genomic distribution is discussed in the light of the available models on transposable element dynamics within inbred genomes, also taking into account SINE role as drivers of genetic diversity in counteracting inbreeding depression. PMID- 19904484 TI - Cytological and physiological changes in orthodox maize embryos during cryopreservation. AB - Cytological and physiological changes during cryopreservation were studied in maize embryos at 35 days after pollination (DAP). Both dehydration and freezing caused cytological damage, such as plasmolysis, swelled mitochondria, increased heterochromatin, and nuclear shrinkage. Dehydration alone slightly impaired plasma membrane integrity while a drastic increase in electrolyte leakage was observed after freezing of embryos with moisture content above 23%. Damage to cellular ultrastructure and plasmalemma integrity was negatively related to moisture content in unfrozen embryos and positively related in frozen embryos. The pattern of changes in activity of antioxidant enzymes differed from one another during dehydration and/or freezing-thawing treatment. Dehydration increased activity of ascorbate peroxidase (APX) and glutathione reductase (GR) but decreased activity of superoxide dismutase (SOD) and dehydroascorbate reductase (DHAR). Freezing further decreased GR and SOD activity and resulted in extremely low DHAR activity. Embryos at intermediate moisture contents had low catalase (CAT) activity before freezing but highest CAT activity after freeze thaw. Both dehydration and freezing promoted membrane lipid peroxidation which resulted in an approximately threefold increase at most in the malondialdehyde content in postthaw embryos. Changes in viability of postthaw embryos can be closely related to damage in cellular ultrastructure and plasmalemma integrity but directly related neither to antioxidants nor lipid peroxidation levels. PMID- 19904486 TI - Factors affecting the retrieval of famous names. AB - Tests of famous faces are used to study language and memory. Yet, the effect of stimulus properties on performance has not been fully investigated. To identify factors influencing proper name retrieval and to probe stimulus-specific parameters within proper name lexicon, we analysed the results obtained by 300 healthy participants on a test of famous faces that includes 74 personalities. A factor analysis yielded five main factors that were characterized by language (national or foreign names), epoch of peak popularity (current, recent or past) and occupation (politicians, entertainment and sports) of the personalities. Multiple regression analysis showed that participants' education, age and gender accounted for 10-32% of the variance in factor scores. These results indicate that there are variables of the stimulus and participants' that must be taken into account in proper name testing and in designing tests aimed to differentiate age-associated difficulties from cognitive decline. PMID- 19904485 TI - The use of tumour necrosis factor alpha-blockers in daily routine. An Austrian consensus project. AB - To define relevant disease parameters and their respective limits indicating the initiation of TNF-alpha-blockers in individual patients. Subsequently, to analyze retrospectively patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA) or ankylosing spondylitis (AS), who started TNF-alpha inhibition in 2006. Points to consider, regarded relevant for individual treatment decisions as well as their assessment methods, were ascertained by experts' consensus applying the Delphi technique. Subsequently, these parameters' thresholds with respect to the initiation of a TNF-alpha-blocker were identified. Thereafter, the rheumatologists representing 12 centres all over Austria agreed to retrospectively analyze their patients started on a TNF-alpha-blocker in 2006. Experts' opinion regarding disease parameters relevant to initiate TNF-alpha blockers in RA patients only slightly differed from those applied in clinical trials, but the parameters' threshold values were considerably lower. For PsA patients, some differences and for AS patients, considerable differences between experts' opinion and clinical studies appeared, which held also true for decisive parameters' means and thresholds. Six hundred and fifty patients, started on TNF blockers in 2006, could be analyzed retrospectively, 408 RA patients (53.3 years mean, 340 females), 93 PsA patients (48.9 years mean, 59 males) and 149 AS patients AS (42.2 years mean, 108 males), representing approximately 25% of all Austrian patients initiated on a TNF-blocker in this respective year. Far more individualized, patient-oriented treatment approaches, at least in part, are applied in daily routine compared with those derived from clinical trials or recommendations from investigative rheumatologists. PMID- 19904487 TI - Interferon-gamma release assay in the ascites: Early hint for diagnosis of abdominal tuberculosis. AB - We report on a 20-year-old woman with abdominal tuberculosis.Standard microbiological examination of ascites showed no acid-fast bacilli (AFB), and analysis for the Mycobacterium tuberculosis (MTB)-complex DNA by PCR was negative. However,the interferon-gamma release assay (IGRA) of the ascites was positive after specific stimulation with mycobacterial antigens(ESAT-6/CFP 10/TB7.7[p4]), indicating an infection with MTB.The diagnosis of tuberculosis was later confirmed by histology, MTB culture, and PCR analysis of MTB-complex DNA in tissue samples taken during laparoscopy. Thus, the IGRA of ascites may guide the decision to start active treatment for tuberculosis. PMID- 19904488 TI - Impact of a change in antibiotic prophylaxis on total antibiotic use in a surgical intensive care unit. AB - OBJECTIVE: The aim of this study was to evaluate the impact of reducing the length of antibiotic prophylaxis for cerebro spinal shunts on total antibiotic use and key resistant pathogens. METHODS: In January 2004, the use of antibiotic prophylaxis was reduced to a single shot dose with cefuroxime in an intensive care unit (ICU). Prior to this intervention, prophylaxis with second-generation cephalosporins was administered during the entire period of external cerebro spinal fluid (CSF) drainage. The effect on the antibiotic use density (AD: DDD [defined daily doses] per 1,000 patient-days[pd]) was calculated prior to (January 2002-December 2003) and following implementation of the intervention(January 2004-December 2006) by segmented regression analysis of an interrupted time series. Resistance proportions(RP) and resistance densities (RD), defined as resistant pathogen/1,000 pd of methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecalis or E. faecium, third-generation-resistant (3GC) Escherichia coli and Klebsiella pneumoniae, and imipenem-resistant Pseudomonus aeruginosa, were compared by the Fisher's exact test before and after the intervention. RESULTS: Total antibiotic use by 147 DDD/1,000 pd decreased after the intervention when pre-operative prophylaxis was changed into single shot prophylaxis, from an estimated mean of 1,036 DDD/1,000 pd before the intervention to 887DDD/1,000 pd post-intervention. This decrease was primarily due to a significant reduction in the amount of cefuroxime used for prophylaxis. The reduction in total antibiotic consumption was sustainable, and it did not increase again during the next 36 months. The RR and RD of third-generation cephalosporin-resistant E. coli increased after January 2004, whereas the percentage of MRSA significantly decreased. CONCLUSION: Change to single shot prophylaxis along with an ongoing antibiotic stewardship program resulted in a cut-back in total antibiotic use amounting to as much as 15%. It would therefore appear that targeting interventions aimed at reducing antibiotic prophylaxis in surgical ICUs may be very worthwhile. PMID- 19904489 TI - Herpes zoster meningoencephalitis: not only a disease of the immunocompromised? PMID- 19904490 TI - Coverage and predictors of adherence to influenza vaccination among Spanish children and adults with asthma. AB - BACKGROUND: Influenza infections are frequently implicated in the exacerbation of asthma. The aim of this study was to assess vaccination coverage among Spanish children and adults with asthma and to describe the factors associated(predictors) with vaccination, using data from the 2006 Spanish National Health Survey. PATIENTS AND METHODS: Subjects were classified as asthma sufferers if they answered affirmatively to the question, "Has your doctor told you that you (or your child) currently suffer from asthma?" Influenza vaccination status was assessed via the question, "Did you (or your child) have a flu shot in the latest campaign?" The following were analyzed as possible predictors of influenza vaccination: sociodemographic variables;health-related and lifestyle variables; variables linked to the use of healthcare services. RESULTS: A total of 38,329 records of individuals aged over 6 months were analyzed, and of these, 2,337 were classified as asthmatics. Among asthmatics, overall influenza coverage was 32.6% (18.8% among children, and 38% among adults).Whereas the sole variable that predicted a higher coverage among children was parents' negative perception of their child's health, among adults positive predictors included older age, lower educational level, suffering from concomitant chronic disease, and non- or ex-smoker status. CONCLUSIONS: The overall influenza vaccination coverage among subjects who suffer from asthma in Spain is very low. Special attention should be paid to children, younger adults, and smokers, since these subjects have the lowest adherence to vaccine recommendations. Strategies focused on health-care providers and patients must be urgently implemented to improve influenza vaccination coverage among asthma sufferers. PMID- 19904491 TI - Management of hepatitis B virus reactivation in patients with hematological malignancies treated with chemotherapy. AB - INTRODUCTION: Hepatitis B virus (HBV) reactivation is a major cause of morbidity and mortality in patients with hematological malignancies who receive cytotoxic chemotherapy. We have therefore carried out a prospective observational study out to assess the incidence, prevalence, and clinical course ina cohort of these patients. METHODS: HBV and HCV markers and liver function indices were monitored prospectively in 318 consecutive patients(171 males, 147 females; mean age 57 years) with hematological malignancies, who had been referred to the Hematology Division, Perugia University, between October 2005 and March 2007 and followed up for at least 6 months. RESULTS: At diagnosis, 32 patients (10%) had received HBV vaccination; 30 were responders. At least one HBV marker was positive in 70/318 patients (22%): 14 (20%) were HBsAg-positive(HBV surface antigen-positive), 13 (19%) were only anti-HBc positive (antibodies to HB core antigen), and 43(61%)were anti-HBc and anti-HBs positive. Twelve HBsAg+ patients received nucleoside/nucleotide analogs (adefovir [six patients],lamivudine [four], and combined adefovir/lamivudine[two non-responders to lamivudine]). After 6 months of therapy, HBV-DNA was negative and transaminases were normal in nine of these 12 patients (adefovir [six], lamivudina[two], adefovir + lamivudina [one]). Seroreversion was achieved in 3/13 patients (23%) who were only anti-HBc positive;all were on rituximab therapy and received adefovir. Seroreversion was not observed in any of the 43 patients who were anti-HBc- and anti-HBs positive. CONCLUSIONS: Essential to the management of patients with hematological malignancies undergoing chemotherapy are surveillance and prophylaxis of HBV infection together with prompt administration of nucleoside/nucleotide analogs in cases of reactivation and/or seroreversion. PMID- 19904494 TI - Clinical and epidemiological features of tularemia. PMID- 19904493 TI - Streptococcus anginosus (Streptococcus milleri Group) Pyomyositis in a 50-Year Old Man with Acquired Immunodeficiency Syndrome: Case Report and Review of Literature. AB - We describe the first reported case of bacterial pyomyositis of the right thigh caused by Streptococcus anginosus (S. milleri group) in an HIV-infected patient. The clinical presentation was complicated by multiple ring-enhancing lesions detected on magnetic resonance imaging of the brain. Evaluation for central nervous system toxoplasmosis,syphilis, and cryptococcal infection was negative. Aggressive antibiotic therapy directed against S. anginosus and surgical debridement were limb salvaging. Clinicians should considerS. anginosus as a causative pathogen in HIV-associated pyomyositis, particularly in complex presentations. Prompt surgical drainage may minimize complications due to S. anginosus, a pathogen associated with significant sequelae due to its invasive nature. PMID- 19904492 TI - Italian guidelines for the diagnosis and infectious disease management of osteomyelitis and prosthetic joint infections in adults. AB - Bone and joint infections are recognized as difficult-to-treat infections that result in significant morbidity and mortality among patients and increased healthcare costs. This article presents the recommendations for the diagnosis and management of osteomyelitis and prosthetic joint infections in adults developed by Bone and Joint Infections Committee for the Italian Society of Infectious and Tropical Diseases. It contains data published through to November 2007. An evidence-based scoring system that is used by the Infectious Diseases Society of America was applied to treatment recommendations. PMID- 19904495 TI - Adjuvant Gamma Knife radiosurgery following surgical resection of brain metastases: a 9-year retrospective cohort study. AB - Given the potential morbidity of whole brain radiation therapy (WBRT), there has been an increasing trend to defer WBRT and deliver Gamma Knife stereotactic radiosurgery (GKS) to cerebral metastatic lesions. We analyzed our experience delivering GKS to the tumor cavity following surgical resection of brain metastases and compared our results to patients receiving WBRT after surgical resection of a metastatic lesion. We performed a retrospective review of patients undergoing surgical resection of at least one brain metastasis between December 1999 and December 2008. Both univariate and multivariate Cox proportional hazards regression were utilized to analyze the influence of various prognostic factors on survival. Twenty-five patients had a metastatic lesion resected followed by adjuvant GKS to the resection cavity while another 18 had surgical resection followed by WBRT. Aside from a disparity in gender distribution (72% of GKS patients were female while women only constituted 28% of the WBRT group), no significant differences existed between groups. The median survival for patients receiving GKS was 15.00 months as compared to 6.81 months among those receiving WBRT (P = 0.08). Univariate Cox regression analysis identified the number of metastases (HR 1.65, 95% CI 1.07-2.54, P = 0.02) and regional recurrence (RR 5.23, 95% CI 1.78-15.38, P = 0.003) as poor prognostic factors. Multivariate regression analysis showed that regional recurrence (HR 5.17, 95% CI 1.69-15.78, P = 0.004) was again strongly associated with worse survival. Although limited by the retrospective nature of our study and lack of some clinical measures, patients undergoing GKS to the resection cavity had a trend towards longer median survival. PMID- 19904496 TI - A surgically treated case of Lhermitte-Duclos disease with a precise natural history and high uptake of FDG on PET. AB - We describe herein a surgically treated case of Lhermitte-Duclos disease in a 33 year-old man. The clinical presentation seemed typical in terms of symptoms, neurological signs, and neuroimaging. High and heterogenous fluorodeoxyglucose uptake in positron emission tomography study is presented and discussed. Furthermore, we performed volumetric analysis of the tumor with sequential magnetic resonance imaging over the course of 7 years before surgery, making this report the first with a long-term natural history, revealing that this rare disease entity may have a neoplastic nature. PMID- 19904497 TI - Role of genetic polymorphism of estrogen receptor-alpha gene and risk of prostate cancer in north Indian population. AB - Present study depicted the role of polymorphisms in estrogen receptor-alpha gene in association with prostate cancer in north Indian population. The study was performed on 157 cases of prostate cancer, 170 cases of BPH, and 170 healthy Indian males diagnosed with prostate cancer and benign prostatic hyperplasia (BPH) and healthy males as controls. Determination of polymorphism in the ER alpha gene was done by polymerase chain reaction followed by restriction fragment length polymorphism (RFLP) analysis with PvuII and XbaI enzymes. An association was observed between PvuII polymorphism of ER-alpha gene and that of prostate cancer. However, there was no such association with XbaI polymorphism in ER-alpha gene. PMID- 19904498 TI - Side chain: backbone projections in aromatic and ASX residues from NMR cross correlated relaxation. AB - The measurements of cross-correlated relaxation rates between H(N)-N and C(beta) C(gamma) intraresidual and sequential dipolar interactions is demonstrated in ASN, ASP and aromatic residues. The experiment can be used for deuterated samples and no additional knowledge such as Karplus parametrizations is required for the analysis. The data constitutes a new type of information since no other method relates the C(beta)-C(gamma) bond to H(N)-N. Using this method the dominant populations of rotamer states of chi 1 can be readily cross checked provided that phi or psi are known. In addition, dynamics on all timescales can be probed. As opposed to standard dynamics analysis of isolated bonds, the presented observables depend on relative dynamics with an interesting prospect to analyze correlated fluctuations of the two torsion angles phi or psi with chi 1. Experimental rates are compared to single conformer and ensemble representations of GB3 and ubiquitin. In particular, it is found that the recently published ubiquitin ensemble 2k39 improves the agreement obtained for 1UBQ. In general, however, input data restricting ASX and aromatic side chains in structure calculation is sparse highlighting the need for new NMR observables. PMID- 19904499 TI - Rice OsACDR1 (Oryza sativa accelerated cell death and resistance 1) is a potential positive regulator of fungal disease resistance. AB - Rice Oryza sativa accelerated cell death and resistance 1 (OsACDR1) encodes a putative Raf-like mitogen-activated protein kinase kinase kinase (MAPKKK). We had previously reported upregulation of the OsACDR1 transcript by a range of environmental stimuli involved in eliciting defense-related pathways. Here we apply biochemical, gain and loss-of-function approaches to characterize OsACDR1 function in rice. The OsACDR1 protein showed autophosphorylation and possessed kinase activity. Rice plants overexpressing OsACDR1 exhibited spontaneous hypersensitive response (HR)-like lesions on leaves, upregulation of defense related marker genes and accumulation of phenolic compounds and secondary metabolites (phytoalexins). These transgenic plants also acquired enhanced resistance to a fungal pathogen (Magnaporthe grisea) and showed inhibition of appressorial penetration on the leaf surface. In contrast, loss-offunction and RNA silenced OsACDR1 rice mutant plants showed downregulation of defense-related marker genes expressions and susceptibility to M. grisea. Furthermore, transient expression of an OsACDR1:GFP fusion protein in rice protoplast and onion epidermal cells revealed its localization to the nucleus. These results indicate that OsACDR1 plays an important role in the positive regulation of disease resistance in rice. PMID- 19904500 TI - Targeted therapies in thyroid cancer. AB - Differentiated thyroid carcinoma is the most frequent neoplasm of the endocrine system. Although thyroid cancer usually has an excellent prognosis, no therapeutic options are available for patients that develop metastases and are or became resistant to radioiodine therapy. The deeper knowledge of molecular aberrations that characterize tumor growth has provided novel targets in cancer therapy. Several proteins have been implicated as having a crucial role in the carcinogenesis of differentiated thyroid cancer, such as those involved in RET/PTC-RAS-RAF-MAPK pathway. Moreover, vascular aberrations and angiogenesis equilibrium have also been related to tumor growth. The development of new, targeted therapies and their encouraging initial results have opened a hopeful opportunity of treatment for these orphan therapy tumor patients. PMID- 19904501 TI - Effect of fluoride exposure on serum glycoprotein pattern and sialic acid level in rabbits. AB - This study describes the effects of fluoride exposure on the protein profile, glycoprotein pattern, and total sialic acid concentration of serum in rabbits. For this aim; 20 healthy New Zealand rabbits were used. The rabbits were divided into two equal groups each with ten animals according to their weighing: control group and experimental group. The rabbits in control group were given drinking tap water containing 0.29 mg/l sodium fluoride and experimental group received the same tap water to which was added 40 mg/l sodium fluoride for 70 days. Blood samples were taken from each rabbit on day 70. Serum fluoride concentrations were measured by a fluoride-specific ion electrode in serum. The fluoride levels in the serum were found as 18.4 (+/-1.58) microg/L in control and 301.3 (+/-52.18) microg/L in fluoride exposed rabbits. The sialic acid levels were found as 69.2 (+/-0.32) mg/dL in control and 43.4 (+/-0.13) mg/dL in fluoride exposed group. The electrophoretic patterns of serum proteins, glycoproteins, and total sialic acid concentration were determined. Fifteen different protein fractions with molecular weights ranging from 22 to 249 kDa were displayed in the serum protein electrophoretic gel of both groups. The raw concentrations of the protein fractions decreased in fluoride exposed rabbits as compared with the control rabbits. The serum glycoprotein pattern revealed seven major protein bands from 47 to 167 kDa in experimental and control groups. The slight decrease of raw concentration of the protein bands in glycoprotein pattern of serum was observed in fluoride toxication comparing to control. The results suggest that serum TSA determination and serum protein electrophoresis can be used to evaluate prognosis of fluoride exposure as a supplementary laboratory test in combination with clinical and other laboratory findings of fluorosis. PMID- 19904502 TI - Complementary and comparative study on hypoglycemic and antihyperglycemic activity of various extracts of Eugenia jambolana seed, Momordica charantia fruits, Gymnema sylvestre, and Trigonella foenum graecum seeds in rats. AB - In present study, we investigated hypoglycemic and antihyperglycemic potential of five extracts (water, ethanol, methanol, hexane, and chloroform) of four plants (i.e., seeds of Eugenia jambolana, fruits of Momordica charantia, leaves of Gymnema sylvestre, and seeds of Trigonella foenum graecum) alone and/or in combination with glimepiride in rats. Ethanol extract of E. jambolana, water extract of M. charantia, ethanol extract of G. sylvestre, and water extract of T. graecum exhibited highest hypoglycemic and antihyperglycemic activity (most active) in rats among all the extracts, while hexane extracts exhibited least activities. Most active extracts were further studied to dose-dependent (200, 100, and 50 mg/kg body weight (bw)) hypoglycemic and antihyperglycemic effects alone and in combination with glimepiride (20, 10, and 5 mg/kg bw). The combination of most active extracts (200 mg/kg bw) and lower dose of glimepiride (5 mg/kg bw) showed safer and potent hypoglycemic as well as antihyperglycemic activities without creating severe hypoglycemia in normal rats, while higher doses (200 mg/kg bw of most active extracts, and 10 and 20 mg/kg bw of glimepiride) were generated lethal hypoglycemia in normal rats. From this study, it may be concluded that the ethanol extract of E. jambolana seeds, water extract of M. charantia fruits, ethanol extract of G. sylvestre leaves, and water extract of T. graecum seeds have higher hypoglycemic and antihyperglycemic potential and may use as complementary medicine to treat the diabetic population by significantly reducing dose of standard drugs. PMID- 19904503 TI - Visualizing dynamical neural assemblies with a fuzzy synchronization clustering analysis. AB - Phase synchrony has been proposed as a possible communication mechanism between cerebral regions. The participation index method (PIM) may be used to investigate integrating structures within an oscillatory network, based on the eigenvalue decomposition of matrix of bivariate synchronization indices. However, eigenvector orthogonality between clusters may result in categorization difficulties for hub oscillators and pseudoclustering phenomenon. Here, we propose a method of fuzzy synchronization clustering analysis (FSCA) to avoid the constraint of orthogonality by combining the fuzzy c-means algorithm with the phase-locking value. Following mathematical derivation, we cross-validated the FSCA and the PIM using the same multichannel phase time series of event-related EEG from a subject performing a working memory task. Both clustering methods produced consistent findings for the qualitatively salient configuration of the original network-illustrated here by a visualization technique. In contrast to PIM, use of common virtual oscillatory centroids enabled the FSCA to reveal multiple dynamical neural assemblies as well as the unitary phase information within each assembly. PMID- 19904504 TI - Pegylated alpha interferon 2B plus ribavirin in the treatment of HCV genotype 4 infection. AB - OBJECTIVE: To evaluate safety and efficacy of Peg-INF combined with ribavirin for genotype 4 infected children. METHODS: Seven children were included, five were infected parentrally, One vertically and one had both exposures. Clinical and laboratory evaluation were undertaken as well as quantitative PCR for HCV RNA before therapy at, 12, 24 and 52 weeks during treatment and one year after therapy. Liver biopsy was performed before and at the end of therapy. Four children had low and three had moderate viremia. RESULTS: At twelve weeks, two children (28.6%) lost viremia. Another child lost viremia at 52 weeks. ETR was 42.9%. During follow up one relapsed, thus SVR was 28.6%. Children with SVR were the youngest, their mean duration of infection was 4.5 vs 12.7 years in the others. Side effects of both INF & ribavirin was mild, required no reduction in doses. CONCLUSION: Combination therapy of peg interferon-alpha with ribavirin is well tolerated in children and adolescents studied. PMID- 19904505 TI - Etiology of upper gastrointestinal bleeding in young children. AB - OBJECTIVE: Evaluation of children younger than two years old admitted to a pediatric emergency department with upper gastrointestinal bleeding. METHODS: Thirty four (34) children aged < 2 years with upper gastrointestinal bleeding were studied. The demographic features, etiologies, laboratory and endoscopic findings and treatment procedures were evaluated retrospectively. RESULTS: The study consisted of 34 patients (73.5 % male, 26.5 % female) with a median age 12.3 months (1.5-24 months). 30 patients (88 %) had an underlying disease. 21 patients (56 %) had a history of aspirin or nonsteroidal anti-inflammatory drugs intake. Endoscopic findings were pathological in 85% of patients. CONCLUSION: The incidence of upper gastrointestinal bleeding in children aged < 2 years due to antipyretics must be taken into consideration although most of them may also have an underlying disease. PMID- 19904506 TI - Detection of fetal malnutrition using "CAN score". AB - OBJECTIVE: Use of clinical assessment of nutrition status (CAN) score to assess the prevalence of fetal malnutrition among term newborns and to compare other anthropometric criteria used to assess fetal growth. METHODS: Prospective study of 529 term healthy newborns assessed using CAN score. Complete anthropometric assessment and determination of weight for gestation was done. Using CAN score as a standard, the usefulness of birth weight, weight for gestation, length, head circumference (HC), mid arm circumference (MAC), MAC/HC ratio and Ponderal index to assess fetal nutrition was determined. RESULTS: With a cut off value of 25, CAN score identified 148 (27.97%) malnourished neonates. 4% of appropriate for gestational age neonate were malnourished and 42.9 % of small for gestational age neonates were well nourished. When CAN score was taken as a standard, weight for gestation and MAC/HC had the highest sensitivity to identify malnourished neonates (92.5% & 90.5%). CONCLUSION: CAN score identifies malnourished neonates which can be missed by other methods and identifies well nourished neonates classified as growth retarded by other methods. PMID- 19904507 TI - Can the special care neonatal unit admission cut-off be lowered down to 1500g babies? AB - OBJECTIVE: The study was conducted to find out whether there is any significant difference in outcome if low birth weight babies weighing between 1500g and 1800g are managed by keeping them with their mothers, i.e., without special care neonatal unit (SCNU) admission. METHODS: It was a retrospective study for which data was collected from past medical record section of 6 months duration from 01.07.06 to 31.12.06, Medical College, Kolkata. The subject of the study was babies born with weight between 1500g and 2000g, divided into 2 groups. Group A representing babies born between weight 1500g and 1800g, groups B representing babies born between 1801g and 2000g. The groups were compared with regard to four variable namely average material age, sex of the babies, singleton or twin pregnancy, mode of delivery and gestational maturity. Test of one proportion was used for statistical analysis of outcome. RESULTS: Total number of live born babies in group A were 198 and in group B 223. Two group were comparable with respect to average maternal age (23.7 yr), sex distribution, singleton or twin pregnancy and number of cesarian section or vaginal delivery and proportion of small for gestational age (SGA) babies. In both the group 13 babies required SCNU admission after being given to their mothers in the postnatal ward. No significant difference in outcome was observed between the groups. CONCLUSION: We conclude that the recommended guidelines for giving institutional care to babies below 1800g may be lowered down to 1500g. However, more babies should be evaluated prospectively, over a longer duration of time, before changing the standard guidelines. PMID- 19904508 TI - Childhood multiple sclerosis mimicking acute disseminated encephalomyelitis. AB - Multiple sclerosis is a demyelinating disease affecting people in early to middle adulthood and is uncommon in childhood. Here we report a patient of multiple sclerosis who presented much before 10 years of age with features suggestive of acute disseminated encephalomyelitis and altered sensorium which is quite different from the features of multiple sclerosis in adults. Diagnosis in this patient was made on the basis of predominant white matter involvement with clinically remitting and relapsing course of the disease on follow up over 3 1/2 years, substantiated by finding multiple areas of demyelinating plaques in magnetic resonance imaging, while visual evoked potential and cerebrospinal fluid study supported demyelination. Hence, children with initial diagnosis of acute disseminated encephalomyelitis need long term follow up as they may develop multiple sclerosis. PMID- 19904509 TI - Pleuropulmonary blastoma in an infant. AB - Pleuropulmonary blastomas (PPB) are rare and highly aggressive tumors. Herein, we report an infantile case of type III PPB. A 9-mth-old boy presented to our unit with a history of cough and tachypnea for 2 days. Chest computed tomography revealed a mass in the left upper lobe, emphysema in the left upper lung, and mediastinum and heart shifted towards the right. The mass was removed completely by left upper lobectomy and histology confirmed diagnosis of type III PPB. The immature blastematous tissue was positive for vimentin while benign epithelium was positive for epithelial membrane antigen and cytokeratin. No lymph nodule metastasis was found in the 7 lymph nodules obtained from the hilum of the lung near the tumor. Currently, the patient is under close follow-up and is doing well. PMID- 19904510 TI - Treatment of autonomous ovarian follicular cyst with long-term anastrozole therapy. AB - Functional follicular ovarian cysts are frequently reported in girls with peripheral precocious puberty (PP). These cysts are usually self-limiting and resolve spontaneously. Several drugs like antiestrogens (tamoxifen) and new aromatase inhibitors are seldom used for treatment. Here we report a girl with peripheral PP who presented with unilateral enlargement of the ovary and a recurrent autonomous ovarian cyst. No skin pigmentation or bone anomaly was noted. The patient was successfully treated with anastrozole, a highly selective aromatase inhibitor. No adverse reaction was noted. Anastrozole is a safe and tolerable drug especially used to suppress estrogen action. PMID- 19904511 TI - Acute bilateral vision loss as the presenting feature of subacute sclerosing panencephalitis. AB - A 12-yr-old boy with an atypical presentation of subacute sclerosing panencephalitis (SSPE) is described. Bilateral macular chorioretinitis preceded the neurological symptoms by 3 weeks. Both visual and neurological features had an acute onset. Clinicians need to be aware that macular chorioretinitis in a child may be the heralding feature of SSPE. PMID- 19904512 TI - Shaken baby syndrome. AB - A 35-day-old male infant with presumed shaken baby syndrome is reported. This first born child to mother educated upto middle school and father tailor by occupation was brought from a remote village 180 kms away from JIPMER. Poor feeding, focal clonic seizures were the initial symptoms. The fundus examination revealed fresh preretinal and vitreous hemorrhages. CT Brain showed right sided subdural hemorrhage with subarachnoid extension and midline shift. He had a normal platelet count and coagulation profile. The sensorium deteriorated and infant expired despite adequate ventilatory support. PMID- 19904513 TI - Pediatric disorders of sex development. AB - The management of disorders of sexual differentiation (DSD) involves a multidisciplinary approach. The main aim of analysis was to study the phenotype karyotype correlation in North Indian children with DSD. The records of pediatric DSD were retrieved and characteristics noted. Of total of 58 children, 43 (74.1%) and 10 (17.2%) were raised as males and females respectively. The mean age at presentation was 31.3+/-9 months. The karyotype was 46XY in 45 (77.6%) and 46XX in 12 (20.7%). CAH was commonest cause of DSD (36.2%), followed by gonadal dysgenesis. Of the 15 patients of 46 XY CAH, there were 5 with 17-alpha hydroxylase deficiency, 2 with 3-beta HSD deficiency and one case of lipoid adrenal hyperplasia. There was an excess of genetic males, possibly due to prevalent socio-cultural factors and gender bias favoring males. There is a need to improve the diagnostic facilities and incorporate a team approach in management of DSD. PMID- 19904514 TI - Childhood Kikuchi-Fujimoto disease. AB - To investigate the clinical features of Kikuchi-Fujimoto disease (KFD) and the relationship between viral infection and this disease in children will be better as Kikuchi-Fujimoto disease (KFD) is a lymphadenopathy. The aim of study is to investigate the clinical features of KFD and the relationship. The age, gender, clinical features and aetiopathogenesis of 36 Chinese children with FKD were reviewed, and the viral antigens were detected. Mean age was 10.1 +/- 2.8 yr with a male to female ratio of 1.8:1. Fever and lymphadenopathy were the most common complaints, noted in 23 and all cases respectively. Skin rash and hepatosplenomegaly were also noted. Leukopenia, anemia, thrombocytopenia and raised ESR were noted in 21, 6, 4 and 31 cases respectively. Epstein-Barr virus (EBV) IgM and IgG was positive in 1 and 24 of 29 cases respectively. Antigens of EBV and herpes simplex virus 2 (HSV 2) were found in the biopsy tissue section from 2 and 1 case respectively. Autoantibodies were noted in 3 of 15 cases. Steroid hormones were administrated for 19 cases with good efficacy. These results imply that children with lymphadenopathy and/or fever may have KFD and thus excisional biopsy of lymph nodes should be performed earlier on. A hyperimmune reaction of immune cells to EBV and HSV2 may play a role in the pathology of KFD. PMID- 19904515 TI - Complications during enalapril and diuretic therapy for congestive cardiac failure. PMID- 19904516 TI - Spontaneous Renal Vein Thrombosis with anti-thrombin III deficiency. PMID- 19904517 TI - Tuberculosis co-infection in HAART naive HIV infected children - an Indian study. PMID- 19904518 TI - Occult neurovesical dysfunction with anorectal malformations. PMID- 19904519 TI - Carpenter's syndrome: a rare craniofacial dysmorphic syndrome. PMID- 19904520 TI - Gemtuzumab ozogamicin-induced long-term remission in a woman with acute myelomonocytic leukemia and bone marrow relapse following allogeneic transplantation. AB - A 56-year-old woman with acute myelomonocytic leukemia underwent myeloablative allogeneic hematopoietic stem cell transplantation (allo-SCT) from a matched unrelated donor in her first complete remission (CR). Veno-occlusive disease (VOD) prophylaxis consisted of low-dose heparin and ursodeoxycholic acid. Graft versus-host disease (GVHD) prophylaxis comprised tacrolimus and short-term methotrexate. On day 14, VOD developed, but gradually resolved with supportive therapy. On day 58, she showed grade II acute GVHD, but this resolved spontaneously. On day 140, she developed hematological relapse with 40.2% marrow infiltration of CD33-positive blasts. Following the discontinuation of tacrolimus, gemtuzumab ozogamicin (GO) was administered. After GO administration, the patient exhibited mild VOD and severe pancytopenia with a sustained high fever for 6 weeks without evident infection. Bone marrow examination revealed severe hypoplastic marrow with 1.3% blasts 4 weeks after GO administration. Although transfusion-dependent pancytopenia persisted for 8 months after GO administration, bone marrow examination revealed the recovery of normal hematopoietic cells with 0.8% blasts. The patient has remained in CR with incomplete blood count recovery for 7 years following GO administration. Although the standard treatment for acute myeloid leukemia relapse after allo-SCT still remains to be established, GO may be a promising option. PMID- 19904521 TI - Africanized honeybees are slower learners than their European counterparts. AB - Does cognitive ability always correlate with a positive fitness consequence? Previous research in both vertebrates and invertebrates provides mixed results. Here, we compare the learning and memory abilities of Africanized honeybees (Apis mellifera scutellata hybrid) and European honeybees (Apis mellifera ligustica). The range of the Africanized honeybee continues to expand, superseding the European honeybee, which led us to hypothesize that they might possess greater cognitive capabilities as revealed by a classical conditioning assay. Surprisingly, we found that fewer Africanized honeybees learn to associate an odor with a reward. Additionally, fewer Africanized honeybees remembered the association a day later. While Africanized honeybees are replacing European honeybees, our results show that they do so despite displaying a relatively poorer performance on an associative learning paradigm. PMID- 19904522 TI - Prediction of maize single-cross hybrid performance: support vector machine regression versus best linear prediction. AB - Accurate prediction of the phenotypic performance of a hybrid plant based on the molecular fingerprints of its parents should lead to a more cost-effective breeding programme as it allows to reduce the number of expensive field evaluations. The construction of a reliable prediction model requires a representative sample of hybrids for which both molecular and phenotypic information are accessible. This phenotypic information is usually readily available as typical breeding programmes test numerous new hybrids in multi location field trials on a yearly basis. Earlier studies indicated that a linear mixed model analysis of this typically unbalanced phenotypic data allows to construct epsilon-insensitive support vector machine regression and best linear prediction models for predicting the performance of single-cross maize hybrids. We compare these prediction methods using different subsets of the phenotypic and marker data of a commercial maize breeding programme and evaluate the resulting prediction accuracies by means of a specifically designed field experiment. This balanced field trial allows to assess the reliability of the cross-validation prediction accuracies reported here and in earlier studies. The limits of the predictive capabilities of both prediction methods are further examined by reducing the number of training hybrids and the size of the molecular fingerprints. The results indicate a considerable discrepancy between prediction accuracies obtained by cross-validation procedures and those obtained by correlating the predictions with the results of a validation field trial. The prediction accuracy of best linear prediction was less sensitive to a reduction of the number of training examples compared with that of support vector machine regression. The latter was, however, better at predicting hybrid performance when the size of the molecular fingerprints was reduced, especially if the initial set of markers had a low information content. PMID- 19904523 TI - Genetic analyses of the host-pathogen system Turnip yellows virus (TuYV)-rapeseed (Brassica napus L.) and development of molecular markers for TuYV-resistance. AB - The aphid transmitted Turnip yellows virus (TuYV) has become a serious pathogen in many rapeseed (Brassica napus L.) growing areas. Three-years' field trials were carried out to get detailed information on the genetics of TuYV resistance derived from the resynthesised B. napus line 'R54' and to develop closely linked markers. F(1) plants and segregating doubled-haploid (DH) populations derived from crosses to susceptible cultivars were analysed using artificial inoculation with virus-bearing aphids, followed by DAS-ELISA. Assuming a threshold of E (405) = 0.1 in ELISA carried out in December, the results led to the conclusion that pre-winter inhibition of TuYV is inherited in a monogenic dominant manner. However, the virus titre in most resistant lines increased during the growing period, indicating that the resistance is incomplete and that the level of the virus titre is influenced by environmental factors. Bulked-segregant marker analysis for this resistance locus identified two closely linked SSR markers along with six closely linked and three co-segregating AFLP markers. Two AFLP markers were converted into co-dominant STS markers, facilitating efficient marker-based selection for TuYV resistance. Effective markers are particularly valuable with respect to breeding for TuYV resistance, because artificial inoculation procedures using virus-bearing aphids are extremely difficult to integrate into practical rapeseed breeding programs. PMID- 19904524 TI - Characterization of an O-desmethylangolensin-producing bacterium isolated from human feces. AB - A bacterium that converted daidzein to O-desmethylangolensin was isolated from the feces of healthy humans. It was an obligately anaerobic, nonsporeforming, nonmotile and Gram-positive rod. The isolate used glucose, sucrose, raffinose, maltose, and fructose as carbon sources. It did not hydrolyze gelatin, esculin, or starch. The strain was urease, acid phosphatase, and arginine dihydrolase positive. It was catalase, oxidase, H(2)S, and indole negative. The major products of glucose fermentation were butyrate and lactate. Its mol% G+C was 51.2. The major cellular fatty acids were C(16:0) DMA, C(16:0), and C(16:0) aldehyde. The structural type of cell wall peptidoglycan was suggested to be A1gamma. The isolate was susceptible to beta-lactam, cefem, and macrolide antibiotics and resistant to aminoglycoside and quinolone antibiotics. The bacterium was related to Eubacterium ramulus ATCC29099(T), Eubacterium rectale ATCC33656(T), and species of the genus Roseburia, but the highest 16S rRNA gene similarity to these described species was only 94.4%, consistent with its being classified as a novel genus. Based on the above, the isolate, named strain SY8519, was identified as belonging to a novel genus in the Clostridium rRNA cluster XIVa. PMID- 19904525 TI - Dectin-1 and inflammation-associated gene transcription and expression in mouse lungs by a toxic (1,3)-beta-D glucan. AB - The form of (1-3)-beta-D glucan found in the cell walls of the anamorphic Trichocomaceae that grow on damp building materials is considered to have potent toxic and inflammatory effects on cells of the respiratory system. It is also considered to have a potential role in the development of non-allergenic respiratory health effects. While human studies involving experimental exposures all point to the inflammatory potential of pure curdlan, a linear (1-3)-beta-D glucan in a triple helix configuration, animal experiments result in conflicting conclusions concerning the inflammatory potency of this glucan. However, because mice appear to be a better model than guinea pigs for exploring the respiratory effects of curdlan and because molecular mechanisms associated with this glucan remain largely unknown, we conducted further work to clarify the role of curdlan on the inflammatory response using our mouse model of lung disease. This study used in situ hybridization (ISH) to probe dectin-1 mRNA transcription with a digoxigenin-labeled cDNA probe, with reverse transcription (RT)-PCR based arrays used to measure inflammation gene and receptor transcriptional responses. Also, immunohistochemistry (IHC) was used to probe dectin-1 as well as anti-mouse Ccl3, Il1-alpha, and TNF-alpha expression to evaluate dose and time-course (4 and 12 h) postexposure (PE) response patterns in the lungs of intratracheally instilled mice exposed to a single 50 mul dose of curdlan at 10(-7), 10(-8), 10(-9), and 10(-10) M/animal (=4 mug to 4 ng curdlan/kg lung wt). Dectin-1 mRNA transcription and expression was observed in bronchiolar epithelium, alveolar macrophages (AMs), and alveolar type II cells (ATIIs) of lungs exposed to 4 mug to 40 ng curdlan/kg lung wt, at both time points. Compared to controls, array analysis revealed that 54 of 83 genes assayed were significantly modulated by curdlan. mRNA transcription patterns showed both dose and time dependency, with highest transcription levels in 10(-7) and 10(-8) M treatment animals, especially at 4-h PE. Nine gene mRNA transcripts (Ccl3, Ccl11, Ccl17, Ifng, Il1alpha, Il-20, TNF alpha, Tnfrsf1b, and CD40lg) were significantly expressed at all doses suggesting they may have a central role in immunomodulating curdlan exposures. IHC revealed Ccl3, Il1-alpha, and TNF-alpha expression in bronchiolar epithelium, AMs and ATIIs illustrate the important immunomodulatory role that these cells have in the recognition of, and response to glucan. Collectively, these results confirm the inflammatory nature of curdlan and demonstrate the complex of inflammation associated gene responses induced by (1-3)-beta-D glucan in triple helical forms. These observations also provide a biological basis for the irritant and inflammatory response to curdlan observed in humans and animals in experimental studies. PMID- 19904526 TI - Protective effect of verbascoside in activated C6 glioma cells: possible molecular mechanisms. AB - The glycosylated phenylpropanoid verbascoside (VB), isolated from cultured cells of the medicinal plant Syringa vulgaris (Oleaceae), has previously been characterized as an effective scavenger of biologically active free radicals and an inhibitor of lipid peroxidation. The aim of the present study was to evaluate in a rat glioma cell line (C6) the effect of VB biotechnologically produced by S. vulgaris plant cell cultures in the regulation of the inflammatory response. We used a model of central nervous system inflammation induced by bacterial endotoxin/cytokine (lipopolysaccharide (LPS)/interferon (IFN)-gamma, 1 microg/ml and 100 U/ml, respectively). Our results show that the treatment with LPS/IFN gamma for 24 h elicited the induction of inducible nitric oxide synthase (iNOS) activity as determined by NO(x) accumulation in the culture medium. Preincubation with VB (10-100 microg/ml) abrogated the mixed cytokine-mediated induction of iNOS. The effect was concentration-dependent. Our studies also showed an inhibitory effect of VB on neuronal nitric oxide synthase expression. Moreover, Western blot analysis showed that this glycoside prevents specifically the activation of the proinflammatory enzyme cyclooxygenase (COX)-2 in glioma cells without simultaneous inhibition of COX-1 enzyme. Moreover, we found that VB reduced the expression of proinflammatory enzymes in LPS/IFN-gamma through the inhibition of the activation of nuclear factor kappa B and mitogen-activated protein kinase signaling pathway. The mechanisms underlying in vitro the neuroprotective properties of VB involve modulation of transcription factors and consequent altered gene expression, resulting in downregulation of inflammation. These findings provide support that VB may provide a promising approach for the treatment of oxidative-stress-related neurodegenerative diseases. PMID- 19904527 TI - Apoptosis induction by doxazosin and other quinazoline alpha1-adrenoceptor antagonists: a new mechanism for cancer treatment? AB - Doxazosin and related, quinazoline-based alpha(1)-adrenoceptor antagonists can induce apoptosis in prostate and various other normal, benign, smooth muscle, endothelial and malignant cells. Such apoptosis-inducing effects occur independently of alpha(1)-adrenoceptor antagonism and typically require much high concentrations than those required for receptor occupancy. Several studies have invested efforts towards the elucidation of the molecular mechanisms underlying doxazosin-induced apoptosis. These include various tumor cells, cardiomyocytes, endothelial cells and bladder smooth muscle cells. While the high concentrations of doxazosin required to induce apoptosis challenge the use of this and related drugs for clinical optimization of apoptosis induction, such quinazoline structure may represent chemical starting points to develop more potent apoptosis inducing agents free of alpha(1)-adrenoceptor antagonistic action and suitable for cancer treatment with minimal and well-tolerated side effects. PMID- 19904528 TI - Memory-guided saccade processing in visual form agnosia (patient DF). AB - According to Milner and Goodale's model (The visual brain in action, Oxford University Press, Oxford, 2006) areas in the ventral visual stream mediate visual perception and oV-line actions, whilst regions in the dorsal visual stream mediate the on-line visual control of action. Strong evidence for this model comes from a patient (DF), who suffers from visual form agnosia after bilateral damage to the ventro-lateral occipital region, sparing V1. It has been reported that she is normal in immediate reaching and grasping, yet severely impaired when asked to perform delayed actions. Here we investigated whether this dissociation would extend to saccade execution. Neurophysiological studies and TMS work in humans have shown that the posterior parietal cortex (PPC), on the right in particular (supposedly spared in DF), is involved in the control of memory-guided saccades. Surprisingly though, we found that, just as reported for reaching and grasping, DF's saccadic accuracy was much reduced in the memory compared to the stimulus-guided condition. These data support the idea of a tight coupling of eye and hand movements and further suggest that dorsal stream structures may not be sufficient to drive memory-guided saccadic performance. PMID- 19904529 TI - Fitts's Law violation and motor imagery: are imagined movements truthful or lawful? AB - Fitts's Law for the timing of targeted movements states that, when target width is held constant, movement time (MT) will increase as the travelled distance increases. Even imagined movements, mentally simulated actions without actual actions, obey Fitts's Law. Recently, a violation of Fitts's Law has been reported; when targets occur in a structured array, MT to the farthest target is shorter than that predicted by Fitts's Law. We conducted two experiments to determine if the violation also occurs for imagined movements. Results showed a close correspondence between real and imaginary MTs across target locations, including the Fitts's violation for the farthest target. We conclude that the violation of Fitts's Law occurs in motor imagery and that the locus of the violation is in the planning stage of action. PMID- 19904532 TI - Identification and characterization of a HER-2/neu epitope as a potential target for cancer immunotherapy. AB - Our aim is to develop peptide vaccines that stimulate tumor antigen-specific T lymphocyte responses against frequently detected cancers. We describe herein a novel HLA-A*0201-restricted epitope, encompassing amino acids 828-836 (residues QIAKGMSYL), which is naturally presented by various HER-2/neu (+) tumor cell lines. HER-2/neu(828-836), [HER-2(9(828))], possesses two anchor residues and stabilized HLA-A*0201 on T2 cells in a concentration-dependent Class I binding assay. This peptide was stable for 3.5 h in an off-kinetic assay. HER-2(9(828)) was found to be immunogenic in HLA-A*0201 transgenic (HHD) mice inducing peptide specific and functionally potent CTL and long-lasting anti-tumor immunity. Most important, using HLA-A*0201 pentamer analysis we could detect increased ex vivo frequencies of CD8(+) T-lymphocytes specifically recognizing HER-2(9(828)) in 8 out of 20 HLA-A*0201(+) HER-2/neu (+) breast cancer patients. Moreover, HER 2(9(828))-specific human CTL recognized the tumor cell line SKOV3.A2 as well as the primary RS.A2.1.DR1 tumor cell line both expressing HER-2/neu and HLA-A*0201. Finally, therapeutic vaccination with HER-2(9(828)) in HHD mice was proven effective against established transplantable ALC.A2.1.HER tumors, inducing complete tumor regression in 50% of mice. Our data encourage further exploitation of HER-2(9(828)) as a promising candidate for peptide-based cancer vaccines. PMID- 19904530 TI - Striatal action-learning based on dopamine concentration. AB - The reinforcement learning hypothesis of dopamine function predicts that dopamine acts as a teaching signal by governing synaptic plasticity in the striatum. Induced changes in synaptic strength enable the cortico-striatal network to learn a mapping between situations and actions that lead to a reward. A review of the relevant neurophysiology of dopamine function in the cortico-striatal network and the machine reinforcement learning hypothesis reveals an apparent mismatch with recent electrophysiological studies. It was found that in addition to the well described reward-related responses, a subpopulation of dopamine neurons also exhibits phasic responses to aversive stimuli or to cues predicting aversive stimuli. Obviously, actions that lead to aversive events should not be reinforced. However, published data suggest that the phasic responses of dopamine neurons to reward-related stimuli have a higher firing rate and have a longer duration than phasic responses of dopamine neurons to aversion-related stimuli. We propose that based on different dopamine concentrations, the target structures are able to decode reward-related dopamine from aversion-related dopamine responses. Thereby, the learning of actions in the basal-ganglia network integrates information about both costs and benefits. This hypothesis predicts that dopamine concentration should be a crucial parameter for plasticity rules at cortico-striatal synapses. Recent in vitro studies on cortico-striatal synaptic plasticity rules support a striatal action-learning scheme where during reward related dopamine release dopamine-dependent forms of synaptic plasticity occur, while during aversion-related dopamine release the dopamine concentration only allows dopamine-independent forms of synaptic plasticity to occur. PMID- 19904533 TI - Does computer-assisted surgery benefit leg length restoration in total hip replacement? Navigation versus conventional freehand. AB - Leg length discrepancy following total hip replacement (THR) can contribute to poor hip function. Abnormal gait, pain, neurological disturbance and patient dissatisfaction have all been described as a result of leg length inequality after THR. The purpose of this study was to determine whether the use of computer navigation in THR can improve limb length restoration and early clinical outcomes. We performed a matched-pair study comparing 48 computer-assisted THR with 48 THRs performed using a traditional freehand alignment method. The same implant with a straight non-modular femoral stem was used in all cases. The navigation system used allowed the surgeon to monitor both acetabular cup placement and all the phases of femoral stem implantation including rasping. Patients were matched for age, sex, arthritis level, pre-operative diagnosis and pre-operative leg length discrepancy. At a minimum follow-up of six months, limb length discrepancy was measured using digital radiographs and a standardised protocol. The number of patients with a residual discrepancy of 10 mm or more and/or a post-operative over-lengthening were measured. The clinical outcome was evaluated using both the Harris Hip Score and the normalised Western Ontario and McMaster Universities (WOMAC) Arthritis Index. Restoration of limb length was significantly better in the computer-assisted THR group. The number of patients with a residual limb length discrepancy greater than 10 mm and/or a post operative over-lengthening was significantly lower. No significant difference in the Harris Hip Score or normalised WOMAC Arthritis Index was seen between the two groups. The surgical time was significantly longer in the computer-assisted THR group. No post-operative dislocations were seen. PMID- 19904534 TI - ABO discrepancy in an elderly patient with IgA kappa-type multiple myeloma. PMID- 19904535 TI - Jumping translocation involving 1q21 during long-term complete remission of acute myeloid leukemia. PMID- 19904536 TI - Fatal gemcitabine-induced pulmonary toxicity in metastatic gallbladder adenocarcinoma. AB - Gemcitabine is a chemotherapy agent that may cause unpredictable side effects. In this report, we describe a fatal gemcitabine-induced pulmonary toxicity in a patient with gallbladder metastatic adenocarcinoma. A 72-year-old patient was submitted to an elective laparoscopic cholecystectomy, and a tubular adenocarcinoma in the gallbladder was incidentally diagnosed. CT scan and ultrasound before the surgery did not show any tumor. After the surgery a Pet scan was positive for a hot-spot in the left colon. The colonic lesion was conveniently removed and the histology evaluation confirmed the diagnosis of adenocarcinoma tubular. The patient was then submitted to three sections of 1,600 mg/m2 of gemcitabine with intervals of 1 week. Three weeks later he developed severe respiratory distress. A helicoidal CT scan showed diffuse and severe interstitial pneumonitis, and lung biopsy confirmed accelerated usual interstitial pneumonia consistent with drug-induced toxicity. The patient presented unfavorable evolution with progressive worsening of respiratory function, hypotension, and renal failure. He died 1 month later in spite of methylprednisolone pulse therapy, large spectrum antimicrobial therapy, and full support of respiratory, hemodynamic and renal systems. Gemcitabine-induced pulmonary toxicity is usually a dramatic condition. Physicians should suspect pulmonary toxicity in patients with respiratory distress after gemcitabine chemotherapy, mainly in elderly patients. PMID- 19904537 TI - Preclinical study of prolonged administration of trastuzumab as combination therapy after disease progression during trastuzumab monotherapy. AB - PURPOSE: The clinical relevance of prolonged trastuzumab administration in combination therapy beyond progressive disease (PD) has been suggested. Here, we examined whether trastuzumab treatment is effective in combination after failing to show antitumor activity as monotherapy in HER2-positive human breast cancer xenograft models. METHODS: We established trastuzumab PD models with HER2 positive breast cancer xenograft models and compared the antitumor activity of trastuzumab in combination with a taxane versus monotherapy with a taxane in the models subsequent to tumor progression under trastuzumab monotherapy. RESULTS: We established trastuzumab PD model using the HER2-positive human breast cancer line MDA-MB-361 and KPL-4 in in vivo. In these models, trastuzumab at the same dose as the initial treatment showed no significant antitumor activity at 3 weeks after start of treatment. Re-inoculated tumor tissues showing PD regained sensitivity to trastuzumab. In the trastuzumab PD models, the HER2 status of the tumor tissues did not decrease. Also, the pAKT level continued to decrease, as with the initial treatment, and IGF-1R was not found to be up-regulated. Instead, differences were observed in the gene-expression profiles of the tumor tissues showing PD. Trastuzumab in combination with G-CSF, which is expected to enhance antibody-dependent cellular cytotoxicity (ADCC), showed significant antitumor activity, even though the single agents alone showed no antitumor activity in the PD model. In the MDA-MB-361 trastuzumab PD model, the combination of trastuzumab with paclitaxel showed significantly more potent antitumor activity compared with paclitaxel or docetaxel monotherapy. In the KPL-4 trastuzumab PD model as well, trastuzumab showed significant antitumor activity in combination with taxanes or capecitabine after PD had developed in response to trastuzumab monotherapy. CONCLUSION: We established in vivo trastuzumab PD models, in which trastuzumab monotherapy ceases to have antitumor activity during the treatment. The mechanisms of PD with trastuzumab are considered to involve both reversible changes in the gene expression profiles in tumor tissues and a decrease of ADCC activity in the host. Our present results demonstrated that trastuzumab showed antitumor activity in combination with taxanes or capecitabine even though it showed no antitumor activity as a monotherapy, suggesting a clinical relevance of treatment with trastuzumab as a combination therapy beyond PD. PMID- 19904538 TI - Phase I clinical trial to determine maximum tolerated dose of oral albendazole in patients with advanced cancer. AB - PURPOSE: Albendazole is a potential anticancer agent that is currently under development for the treatment of cancer. We carried out a dose-finding phase I study of oral albendazole in patients with advanced malignancies. PATIENTS AND METHODS: Thirty-six patients with refractory solid tumors were enrolled. Albendazole was given orally on a day 1-14 of a 3 weekly cycle, starting at 400 mg BD with dose escalation until 1,200 mg BD. Serial blood samples were collected up to 96 h and also on day 8 of cycles 1 and 4. RESULTS: The maximum tolerated dose was 2,400 mg per day (1,200 BD). Myelosuppression was the main dose limiting toxicity. Fatigue and mild gastrointestinal upset were the other major adverse effects. 4 out of 24 assessable patients (16%) had a tumor marker response with a fall of at least 50% from baseline values and another patient had a prolonged period of stable marker response. A decline in plasma vascular endothelial growth factor levels was observed. CONCLUSIONS: Albendazole was well tolerated on the schedule tested in this trial. The results of this study suggest that the recommended dose for further study is 1,200 mg twice daily for 14 days in a 21 day cycle. PMID- 19904539 TI - Asymptotic behavior of a Moran model with mutations, drift and recombination among multiple loci. AB - In this paper, we extend the theoretical treatment of the Moran model of genetic drift with recombination and mutation, which was previously introduced by us for the case of two loci, to the case of n loci. Recombination, when considered in the Wright-Fisher model, makes it considerably less tractable. In the works of Griffiths, Hudson and Kaplan and their colleagues important properties were established using the coalescent approach. Other more recent approaches form a body of work to which we would like to contribute. The specific framework used in our paper allows finding close-form relationships, which however are limited to a set of distributions, which jointly characterize allelic states at a number of loci at the same or different chromosome(s) but which do not jointly characterize allelic states at a single locus on two or more chromosomes. However, the system is sufficiently rich to allow computing, albeit in general numerically, all possible multipoint linkage disequilibria under recombination, mutation and drift. We explore the algorithms enabling construction of the transition probability matrices of the Markov chain describing the process. We find that asymptotically the effects of recombination become indistinguishable, at least as characterized by the set of distributions we consider, from the effects of mutation and drift. Mathematically, the results are based on the foundations of the theory of semi-groups of operators. This approach allows generalization to any Markov-type mutation model. Based on these fundamental results, we explore the rates of convergence to the limit distribution, using Dobrushin's coefficient and spectral gap. PMID- 19904541 TI - Facial palsy in a 2-month-old infant with Kawasaki disease. PMID- 19904542 TI - Perspectives on overactive bladder in the elderly population. AB - INTRODUCTION: Overactive bladder (OAB) represents a disruption in the storage function of the lower urinary tract. This bothersome condition occurs more commonly in the elderly. Since population forecasts predict a worldwide increase in the proportion of people aged over 65 years, it is reasonable to expect that the healthcare burden associated with OAB will also increase. The pathophysiology of OAB in the elderly is thought to be multifactorial, with an abnormality occurring in the nervous supply and/or the structure/function of the urothelium or bladder smooth muscle, leading to bladder hypersensitivity, abnormalities in bladder sensation (urgency) and involuntary detrusor contraction. METHODS: A review of some of the key aspects relating to management of this growing population was undertaken. RESULTS: The potential for an elderly patient to present with a number of concomitant conditions means that careful characterization of their overall status is required before deciding on the most appropriate management option for their urinary tract pathology. Lifestyle interventions and pharmacological agents have shown success in treating OAB in the elderly, but as this patient group often has many concomitant conditions they are more likely to be seen by a non-urology specialist. CONCLUSIONS: It is therefore important to raise awareness of the condition and an appreciation of its impact among healthcare professionals to ensure the most appropriate care. PMID- 19904543 TI - Treatment of cystic craniopharyngioma with phosphorus-32 intracavitary irradiation. AB - PURPOSE: The aim of the study was to evaluate the effect of phosphorus-32 colloid ([(32)P]) intracavitary irradiation on the treatment of patients with cystic craniopharyngiomas. METHODS: Twenty patients with predominantly cystic craniopharyngiomas were admitted from 1981 to 2006. Eleven patients had [(32)P] intracavitary irradiation by stereotactic injection or Ommaya cyst instillation as the primary treatment, and the remaining nine had the same internal irradiation as an adjuvant treatment after tumor resection. A calculated irradiation dose of 400 approximately 500 Gy per once was delivered to the cyst wall. CONCLUSION: The patients were followed up ranging from 36 to 336 months; no operative morbidity or mortality was found from [(32)P] intracavitary irradiation. Fourteen patients (70%) had tumor progression and required further two to four times intracavitary irradiation. All 20 cases achieved tumor shrinkage or stabilization with effective outcome 3-6 months after the last [(32)P] therapy. For patients with cystic craniopharyngioma, [(32)P] administration by stereotactic injection or Ommaya cyst instillation is a safe and helpful option, which could improve the life quality, prolong the life span, and enhance the survival rate of cystic craniopharyngioma patients. PMID- 19904544 TI - Intraoperative neurophysiology of the conus medullaris and cauda equina. AB - PURPOSE: Intraoperative neurophysiological techniques are becoming routine tools for neurosurgical practice. Procedures affecting the lumbosacral nervous system are frequent in adult and pediatric neurosurgery. This review provides an overview of the techniques utilized in cauda and conus operations. METHODS: Two basic methodologies of intraoperative neurophysiological testing are utilized during surgery in the lumbosacral spinal canal. Mapping techniques help identify functional neural structures, namely, nerve roots and their respective spinal levels. Monitoring is referred to as the technology to continuously assess the functional integrity of pathways and reflex circuits. For mapping direct electrical stimulation of a structure within the surgical field and recording at a distant site, usually a muscle is the most commonly used setup. Sensory nerve roots or spinal cord areas can be mapped by stimulation of a distant sensory nerve or skin area and recording from a structure within the surgical field. Continuous monitoring of the motor system is done with motor evoked potentials. These are evoked by transcranial electrical stimulation and recorded from lower extremity and sphincter muscles. Presence or absence of muscle responses are the monitored parameters. To monitor the sensory pathways, sensory potentials evoked by tibial, peroneal, or pudendal nerve stimulation and recorded from the dorsal columns with a spinal electrode or as cortical responses from scalp electrodes are used. Amplitudes and latencies of these responses are measured for interpretation. The bulbocavernosus reflex, with stimulation of the pudendal nerve and recording from the external anal sphincter, is used for continuous monitoring of the reflex circuitry. The presence of absence of this response is the pertinent parameter monitored. Stimulation of individual dorsal nerve roots is used to identify those segments that generate spastic activity and which may be cut during selective dorsal rhizotomy. Electromyographic activity can be continuously observed during surgery, and monitoring concepts developed in cranial nerve surgery may be used in the cauda equina as well. CONCLUSION: A range of intraoperative neurophysiological techniques are available for neurophysiological testing of the neural structures of conus medullaris and cauda equina. PMID- 19904545 TI - Symphysiotomy: a valuable approach in children with prostate rhabdomyosarcoma. AB - Surgery for large prostate rhabdoyasarcoma in children is a challenging procedure. We discussed the value of pubic symphysiotomy in affected patients. The symphysiotomy approach was used in two children with a large rhabdomyosarcoma of the prostate. In each case, the initial exposure was obtained through a lower midline incision, but, due to technical difficulties, resulting from the size of the tumor, surgery was completed via a symphysiotomy approach. In each case, the bladder was preserved and a radical prostatectomy was facilitated by the excellent exposure provided by the symphysiotomy. The patients have been followed for 6 years and 26 months, respectively. Both are tumor free. Neither has developed orthopedic complications. In conclusion, the symphysiotomy approach, for large prostate rhabdomyosarcoma in children, results in an excellent surgical exposure, thus, facilitating the performance of a radical prostatectomy with bladder preservation. Orthopedic complications have not developed throughout the follow up period. PMID- 19904546 TI - Pilot study on the effectiveness of the conventional CROS, the transcranial CROS and the BAHA transcranial CROS in adults with unilateral inner ear deafness. AB - The objective of the present pilot study is to evaluate the effectiveness of three conventional contralateral routing of sound (CROS) hearing aids in adults with unilateral inner ear deafness. The study included tertiary referral center. Ten patients with unilateral inner ear deafness and normal hearing in the contralateral ear were selected to evaluate three different methods of amplification: the CROS hearing aid, the completely in the canal hearing aid and the bone-anchored hearing aid CROS (BAHA). Each of the three hearing aids was tried in a random order for a period of 8 weeks. Audiometric performance, including speech-in-noise, directional hearing and subjective benefit were measured after each trial period, using the APHAB, SSQ and single-sided deafness questionnaire. Sound localization performance was essentially at chance level in all four conditions. Mixed results were seen on the other patient outcome measures that alternated in favor of one of the three CROS devices. After the trial, three patients chose to be fitted with the BAHA CROS and one with the conventional CROS. In conclusion, most of the patients experienced some degree of benefit with each of the three hearing aids. Preference for one of the three hearing aids was independent of the order in which they were tried. It would be worthwhile to formulate selection criteria; still, we recommend that all patients with unilateral inner ear deafness should be offered a trial with at least the BAHA CROS. PMID- 19904547 TI - How many nodes are needed to stage a neck? A critical appraisal. AB - Reliable staging of the neck is an important factor for the estimation of prognosis of head and neck cancer patients. A total of 608 patients with oral squamous cell carcinomas treated from March 1975 to December 2000 were enrolled. In radical neck dissection (RND) group, the number of lymph nodes ranged from 6 to 116; while in selective neck dissection (SND) group, from 1 to 87 (P < 0.001). In SND group, the number of metastatic nodes ranged from 0 to 8 nodes, while in RND group, from 0 to 47 (P < 0.001). The number of dissected lymph nodes correlates with the presence of positive nodes (P = 0.001). In RND group, this correlation is described by the equation Y = -0.0117X (2) + 1.7262X. Factors affecting neck metastasis were number of dissected nodes (P < 0.001), lymphatic embolization (P = 0.044) and neural invasion (P = 0.030). In SND group, this equation is Y = -0.012X (2) + 1.5102X; the number of dissected nodes (P = 0.002) and lymphatic embolization (P = 0.001) were significant for metastasis finding. For patients with tumors at stages I and II, a significant impact on survival and neck recurrence rates were observed. In conclusion, we report the importance of the number of retrieved nodes in likelihood of positive cervical node finding. Node yield is an important factor in oral cancer staging, and, more important, in early stage carcinomas, it is associated with survival and recurrence rates. PMID- 19904548 TI - Chromosomal and telomeric reprogramming following ES-somatic cell fusion. AB - Chromosomal and telomeric reprogramming was assessed in intraspecies hybrids obtained by fusion of embryonic stem (ES) cells and mouse embryonic fibroblasts. Evaluation of the ploidy of ES-somatic hybrids revealed that 21 of 59 clones had a tetraploid DNA profile while the remaining clones showed deviations from the expected profile of fusion between two diploid cells. Microsatellite polymerase chain reaction analysis of four of these clones demonstrated no random loss of somatic chromosome pairs in the ES-somatic cell hybrids. Pluripotential of ES somatic hybrids was assessed by gene expression analysis, antibody staining for Oct4 and SSEA-1 and teratoma formation containing derivatives of the three germ layers. Reprogramming of telomeric maintenance was observed with ES-somatic hybrids showing high telomerase activity and increased telomere lengths. However, we detected no significant increase in the expression of the three critical telomerase subunits: telomerase reverse transcriptase (TERT), telomerase RNA component (TERC), and dyskerin. This indicates that activation of telomerase and telomere maintenance is not reliant on changes in gene expression of TERT, TERC, and dyskerin following ES-somatic cell fusion or sister chromatid recombination and may arise through elimination of negative regulation of telomerase activity. This is the first demonstration of telomere lengthening following cell fusion and offers a new model for studying and identifying new regulators of telomere maintenance. PMID- 19904549 TI - Re-examination of siRNA specificity questions role of PICH and Tao1 in the spindle checkpoint and identifies Mad2 as a sensitive target for small RNAs. AB - The DNA-dependent adenosine triphosphatase (ATPase) Plk1-interacting checkpoint helicase (PICH) has recently been implicated in spindle checkpoint (SAC) signaling (Baumann et al., Cell 128(1):101-114, 2007). Depletion of PICH by siRNA abolished the SAC and resulted in an apparently selective loss of Mad2 from kinetochores, suggesting a role for PICH in the regulation of the Mad1-Mad2 interaction. An apparent rescue of SAC functionality by overexpression of PICH in PICH-depleted cells initially seemed to confirm a role for PICH in the SAC. However, we have subsequently discovered that all PICH-directed siRNA oligonucleotides that abolish the SAC also reduce Mad2 mRNA and protein expression. This reduction is functionally significant, as PICH siRNA does not abolish SAC activity in a cell line that harbors a bacterial artificial chromosome driving the expression of murine Mad2. Moreover, we identified several siRNA duplexes that effectively deplete PICH but do not significantly affect SAC functionality or Mad2 abundance or localization. Finally, we discovered that the ability of overexpressed PICH to restore SAC activity in PICH-depleted cells depends on sequestration of the mitotic kinase Plk1 rather than ATPase activity of PICH, pointing to an underlying mechanism of "bypass suppression." In support of this view, depletion or inhibition of Plk1 also rescued SAC activity in cells harboring low levels of Mad2. This observation suggests that a reduction of Plk1 activity partially compensates for reduced Mad2 levels and argues that Plk1 normally reduces the strength of SAC signaling. Collectively, our results question the role of PICH in the SAC and instead identify Mad2 as a sensitive off target for small RNA duplexes. In support of the latter conclusion, our evidence suggests that an off-target effect on Mad2 may also contribute to explain the apparent role of the Tao1 kinase in SAC signaling. PMID- 19904550 TI - Amniotic fluid aspiration in cases of SIDS. AB - The scope of this study was to evaluate the incidence and the eventual consequences of amniotic fluid aspiration (AFA) in cases of sudden infant death. Cases of sudden infant death syndrome (SIDS; n = 113: 39 females, 74 males; mean age 4.6 months) were compared to a control group of 39 cases of explained death (14 females, 25 males; mean age 5.6 months). In each case, sections of the lung stained with hematoxylin and eosin and with the immunohistochemical reaction 34BE12 specific for cytokeratins were available. The microscope slides were observed at x200 magnification and semi-quantitatively classified into four categories(-, +, ++, and +++). In both groups, rests of amniotic fluid could be observed up to the fourth month of life. The comparison between the two groups did not show any significant difference. In the SIDS group, immunohistochemical reactions with the antibodies CD68, MRP8, MRP14, 27E10, 25F9, CD3, CD20Cy, and CD45R0 were available for the lungs. Twelve cases with AFA were compared to a group of SIDS cases without AFA with similar age and pathological distribution to evaluate whether the presence of amniotic remnants induced inflammatory changes in the lungs. No differences emerged. This study shows that AFA is not a rare event. Even moderate to severe AFA does not necessary cause death. A correlation between AFA and SIDS could not be shown. PMID- 19904551 TI - Haplotype-assisted characterization of germline mutations at short tandem repeat loci. AB - In this study, 98 families with 101 mutations were analyzed in depth in which a mutation had been observed at one of the four loci D3S1358, FGA, ACTBP2, and VWA. To determine the origin (male/female) of the mutation, five to seven polymorphic flanking markers were selected for each locus concerned and used to construct family-specific haplotypes. Additionally, all alleles of the STR system concerned were sequenced. With this duplicate approach, it was possible to identify the mutated structure and/or mutation event in the vast majority of cases. The ratio of one-step to two-step mutations was 100:1. The ratio of paternal to maternal mutations was 76:8. The ratio of gains to losses was 47:50. Also, the mutation rates in two systems, ACTBP2 and VWA, were clearly higher than those given in the literature. PMID- 19904552 TI - Organizational justice, psychological distress, and work engagement in Japanese workers. AB - PURPOSE: To investigate the cross-sectional association between organizational justice (i.e., procedural justice and interactional justice) and psychological distress or work engagement, as well as the mediating roles of other job stressors (i.e., job demands and job control, or their combination, effort-reward imbalance [ERI], and worksite support). METHODS: A total of 243 workers (185 males and 58 females) from a manufacturing factory in Japan were surveyed using a self-administered questionnaire including the Organizational Justice Questionnaire, Job Content Questionnaire, Effort-Reward Imbalance Questionnaire, K6 scale, Utrecht Work Engagement Scale, and other covariates. Multiple mediation analyses with the bootstrap technique were conducted. RESULTS: In the bivariate analysis, procedural justice and interactional justice were significantly and negatively associated with psychological distress; they were significantly and positively associated with work engagement. In the mediation analysis, reward at work (or ERI) significantly mediated between procedural justice or interactional justice and psychological distress; worksite support significantly mediated between procedural justice or interactional justice and work engagement. CONCLUSION: The effects of organizational justice on psychological distress seem to be mediated by reward at work (or ERI) while those regarding work engagement may be mediated by worksite support to a large extent, at least in Japanese workers. PMID- 19904553 TI - Association of heart rate recovery after exercise with indices of obesity in healthy, non-obese adults. AB - We aimed at determining whether body mass index (BMI), waist circumference (WC), and waist-to-hip ratio (WHR) are associated with heart rate recovery (HRR) and to demonstrate which of the three indices of obesity, is the strongest predictor of HRR in apparently healthy non-obese adults. Three hundred and twenty-five subjects aged 18-66 years participated in the study. Anthropometric indices were measured, and subjects performed cycle ergometer exercise at 75-85% maximum heart rate. Heart rate (HR) was measured during the last minute of exercise and in the first minute of post-exercise recovery. A partial correlation test and a multiple linear regression analysis, after adjusting for age and peak oxygen uptake indicated that the best predictors of HRR were BMI in males and WHR in females. The present data suggest that, HRR is independently related to indices of obesity BMI, WC, and WHR and strengthen the usefulness of these anthropometric indices in predicting cardiovascular risks. In addition, the findings suggest that BMI in men and WHR in women best express the relationship between obesity and cardiovascular risks. PMID- 19904554 TI - Hypoxia application in athletes is not doping. PMID- 19904555 TI - Development of an equation to predict muscle volume of elbow flexors for men and women with a wide range of age. AB - The present study examined the age-related changes in muscle thickness (MT) and volume (MV) of elbow flexors and developed a prediction equation of the MV based on the MT applicable to men and women with a wide range of age. The MT and MV were determined from a single ultrasonographic image and multiple magnetic resonance imaging scans, respectively, in 72 men and 75 women aged 19-77 year. As a result of examining the age-MT and age-MV relationships by calculation of partial correlation coefficients with the control variable of gender, MV was decreased with aging whereas the corresponding decline in MT was not significant. The subjects were randomly separated into either a validation (38 men and 42 women) or a cross-validation (34 men and 33 women) group, and a multiple regression equation to estimate MV using not only MT but also upper arm length (L), age and gender as independent variables [MV (cm(3)) = 60.8 x MT (cm) + 6.48 x L (cm) - 0.709 x age (year) + 51.4 x gender (0 women, 1 men) - 187.4] was validated and cross-validated. Thus, the prediction equation for MV of elbow flexors newly developed was shown to be applicable to men and women with a wide range of age. PMID- 19904557 TI - The polyembryonic wasp Copidosoma floridanum produces two castes by differentially parceling the germ line to daughter embryos during embryo proliferation. AB - Eggs of the polyembryonic wasp Copidosoma floridanum undergo a clonal phase of proliferation, which results in the formation of thousands of embryos called secondary morulae and two castes called reproductive and soldier larvae. C. floridanum establishes the germ line early in development, and prior studies indicate that embryos with primordial germ cells (PGCs) develop into reproductive larvae while embryos without PGCs develop into soldiers. However, it is unclear how embryos lacking PGCs form and whether all or only some morulae contribute to the proliferation process. Here, we report that most embryos lacking PGCs form by division of a secondary morula into one daughter embryo that inherits the germ line and another that does not. C. floridanum embryos also incorporate 5-bromo-2' deoxyuridine (BrdU), which allows PGCs and other cell types to be labeled during the S phase of the cell cycle. Continuous BrdU labeling indicated that all secondary morulae cycle during the proliferation phase of embryogenesis. Double labeling with BrdU and the mitosis marker anti-phospho-histone H3 indicated that the median length of the G2 phase of the cell cycle was 18 h with a minimum duration of 4 h. Mitosis of PGCs and presumptive somatic stem cells in secondary morulae was asynchronous, but cells of the inner membrane exhibited synchronous mitosis. Overall, our results suggest that all secondary morulae contribute to the formation of new embryos during the proliferation phase of embryogenesis and that PGCs are involved in regulating both proliferation and caste formation. PMID- 19904556 TI - Extended expression of B-class MADS-box genes in the paleoherb Asarum caudigerum. AB - Asarum caudigerum (Aristolochiaceae) is a paleoherb species that is important for research in origin and evolution of angiosperm flowers due to its basal position in the angiosperm phylogeny. In this study, a subtracted floral cDNA library from floral buds of A. caudigerum was constructed and cDNA arrays by suppression subtractive hybridization were generated. cDNAs of floral buds at different stages before flower opening and of leaves at the seedling stage were used. The macroarray analyses of expression profiles of isolated floral genes showed that 157 genes out of the 612 unique ESTs tested revealed higher transcript abundance in the floral buds and uppermost leaves. Among them, 78 genes were determined to be differentially expressed in the perianth, 62 in the stamens, and 100 genes in the carpels. Quantitative real-time PCR of selected genes validated the macroarray results. Remarkably, APETALA3 (AP3) B-class genes isolated from A. caudigerum were upregulated in the perianth, stamens and carpels, implying that the expression domain of B-class genes in this basal angiosperm was broader than those in their eudicot counterparts. PMID- 19904558 TI - The importance of -460 C/T and +405 G/C single nucleotide polymorphisms to the function of vascular endothelial growth factor A in colorectal cancer. AB - PURPOSE: The present study investigated the functional influence of the single nucleotide polymorphisms (SNPs) -460 C/T and +405 G/C at vascular endothelial growth factor A (VEGF-A), mRNA and protein levels in colorectal cancer (CRC) and normal colorectal tissue. METHODS: Blood and tissue were collected from 113 patients surgically resected for colorectal cancer. SNPs were analysed from genomic DNA by PCR, the VEGF-A gene expression analysis was performed by RT-PCR and protein analysis by ELISA. RESULTS: The T-allele in the -460 C/T SNP and the C-allele in the +405 G/C SNP were associated with significantly lower VEGF-A protein levels in normal colorectal tissue. There were no differences in protein levels in the malignant tissue according to genotypes. No differences were observed at the gene expression levels either. CONCLUSION: The results indicate that the two SNPs have a functional influence on the VEGF-A protein levels in normal colorectal tissue. The possible clinical implications of the findings need further investigation. PMID- 19904559 TI - Effect of bevacizumab in older patients with metastatic colorectal cancer: pooled analysis of four randomized studies. AB - BACKGROUND: Bevacizumab is frequently combined with 5-fluorouracil-based chemotherapy for patients with metastatic colorectal cancer (mCRC). The relative benefit of bevacizumab in older patients has not been widely studied and is of interest. PATIENTS AND METHODS: This retrospective analysis used data from three first-line randomized controlled studies and one second-line randomized controlled study of bevacizumab plus chemotherapy in medically fit (Eastern Cooperative Oncology Group performance status 0 or 1) patients with mCRC. Overall survival (OS) and on-treatment progression-free survival (PFS) were assessed in patients aged <65, > or =65, and > or =70 years. Results were compared using unstratified hazard ratios (HRs). Grade 3-5 adverse events were also assessed. RESULTS: Bevacizumab statistically significantly improved PFS [HR 0.58; 95% confidence interval (CI) 0.49-0.68] and OS (HR 0.85; 95% CI 0.74-0.97) in patients aged > or =65 years; patients aged > or =70 years had similar improvements. Benefits were consistent across the studies, irrespective of setting, bevacizumab dose, or chemotherapy regimen. Increases in thromboembolic events were observed in patients aged > or =65 and > or =70 years in the bevacizumab group compared with the control group, mainly as a result of increases in arterial thromboembolic events. No other substantial age-related increases in grade 3-5 adverse events were observed. CONCLUSIONS: In medically fit older patients, bevacizumab provides similar PFS and OS benefits as in younger patients. PMID- 19904560 TI - Circulating IL-1beta levels, polymorphisms of IL-1B, and risk of cervical cancer in Chinese women. AB - PURPOSE: Long-term human papillomavirus (HPV) infection is a prerequisite for cervical cancer. IL-1beta and IL-1Ra expression levels play an important role in cervical carcinogenesis. Several functional genetic variants in IL1B and IL-RN have been reported to be associated with IL-1beta expression and cancer susceptibility. In the current study, we hypothesized that plasma IL-1beta levels, IL-1B and IL-RN polymorphisms were candidate biomarkers for cervical cancer. METHODS: We measured plasma IL-1beta levels and genotyped IL-1B and IL-RN polymorphisms in a case-control study of 404 cervical cancer cases and 404 controls in Chinese women. RESULTS: The mean plasma IL-1beta levels in cervical cancer cases (42.19 +/- 31.55 pg/ml) was significantly higher than those in controls (34.86 +/- 22.68 pg/ml, P = 0.0002), and plasma IL-1beta levels above the 75% quartiles in controls (IL-1beta > or = 46.94 pg/ml) were associated with a 1.74-fold significantly increased risk of cervical cancer [95% confidence interval (CI), 1.28-2.36], compared with those of lowest quartile. Multivariate logistic regression analyses revealed that the variant genotypes, IL-1B T-31C TC/CC and C-511T CT/TT, were associated with a significantly increased risk of cervical cancer [adjusted odds ratio (OR), 1.60; 95% CI, 1.16-2.21 for -31TC/CC, and adjusted OR, 1.52; 95% CI, 1.10-2.09 for -511CT/TT, respectively), especially among subjects having higher levels of IL-1beta. However, IL-RN VNTR polymorphism was not associated with cervical cancer risk in the current study. Furthermore, the significant differences of IL-1beta concentration between cervical cancer cases and controls were observed only among subjects carrying T-31C or C-511T variant genotypes. CONCLUSION: Functional IL-1B genotypes may modify plasma IL 1beta concentrations to contribute to the etiology of cervical cancer in Chinese women; however, further perspective studies are warranted to test the causal effects of IL-1beta concentration in cervical carcinogenesis. PMID- 19904561 TI - Cervine genotype is the major Cryptosporidium genotype in sheep in China. AB - To identify Cryptosporidium species/genotypes in sheep in China and to elucidate the endemic transmission of cryptosporidiosis, a total of 1,701 fecal samples from five farms in four prefectures in Henan Province (central China) were examined. Eighty-two Cryptosporidium-positive samples were analyzed by polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis of the small subunit (SSU) rRNA gene and PCR analysis of the 60 kDa glycoprotein (gp60) gene, and 41 were further analyzed by DNA sequencing of the PCR products. The SSU rRNA-based PCR identified two Cryptosporidium species and one genotype, including the Cryptosporidium cervine genotype (74/82), Cryptosporidium andersoni (4/82), and Cryptosporidium xiaoi (4/82). The cervine genotype was found in all age groups, C. xiaoi in lambs, and C. andersoni in ewes. There were intragenetic differences in the SSU rRNA gene sequences of the Cryptosporidium cervine genotype and C. xiaoi. No Cryptosporidium parvum was detected by both SSU rRNA- and gp60-based PCR assays. These findings suggest that sheep are a potential source for zoonotic infections of the Cryptosporidium cervine genotype. PMID- 19904563 TI - Promoting evidence-based management of anemia in cancer patients: concurrent and discriminant validity of RESPOND, a web-based clinical guidance system based on the EORTC guidelines for supportive care in cancer. AB - GOAL OF WORK: The goal of this study is to test the validity of RESPOND, a web based decision support system to assess and manage anemia in cancer patients as per the European Organisation for Research and Treatment of Cancer (EORTC) guidelines. The intraclass correlation metrics for the algorithmic definitions were reported previously. Reported here are the concurrent validity, the extent to which clinicians' anemia management is guidelines-congruent when using the system; and discriminant validity, the extent to which clinicians practice in congruence with guidelines when vs. when not using the system. PATIENTS AND METHODS: Hybrid matched design with precohort (retrospective; clinicians not using RESPOND) and postcohort (prospective; clinicians using RESPOND) of anemic patients matched on cancer type and chemotherapy regimen and followed up over 4 months after treatment initiation with erythropoietic proteins (34 patients per cohort; total N = 68). Congruence scores quantified the extent to which anemia management was congruent with the EORTC guidelines (range 0-10). MAIN RESULTS: Hemoglobin (Hb) increased significantly for both cohorts, but the postcohort group showed more rapid rate of Hb increase over time (p < 0.006), higher Hb by visit 4 (p = 0.007), and greater Hb increase by visit 4 (p = 0.006). Concurrent validity was high with mean postcohort congruence scores of 8.18 +/- 1.38. Discriminant validity was inferred from statistically significant differences in mean congruence scores between cohorts (p < 0.001) and from the postcohort having odds ratios of 3.64 for patients to reach Hb >or= 11 g/dL and 2.91 to achieve Hb >or= 12 g/dL. CONCLUSIONS: RESPOND, a validated computerized clinical guidance system with an incremental effect beyond the pharmacotherapeutic effect of erythropoietic proteins, offers clinicians accurate and safe guidance in managing anemia in cancer patients. PMID- 19904562 TI - Prediction of outcome of patients with metastatic breast cancer: evaluation with prognostic factors and Nottingham prognostic index. AB - GOALS OF WORK: The purpose of this study is to analyze the survival rate of patients with metastatic breast cancer and to evaluate the outcome of these patients using prognostic factors and Nottingham prognostic index. MATERIALS AND METHODS: From February 1992 to August 2008, 135 patients with metastatic breast cancer were treated at the Changhua Christian Hospital. In these patients, we evaluated the significance of the following factors in predicting the survival rate after the occurrence of metastasis: age, initial stage at primary diagnosis, histological grade, Karnofsky performance status (KPS), estrogen receptor (ER), progesterone receptor status, human epidermoid growth factor receptor 2 overexpression status, number of axillary lymph node metastasis, history of adjuvant radiotherapy and/or chemotherapy, disease-free interval, status of local recurrence, status of various sites of distant metastases, number of distant metastases, and Nottingham prognostic index. MAIN RESULTS: The 1-, 2-, and 5-year survival rates were 53.3%, 25.2%, and 1.5%, respectively. In the univariate analysis, KPS, histological grade, ER status, initial stage at primary diagnosis, number of axillary lymph node metastasis, liver metastasis, disease-free interval, first-/second-/third-line chemotherapy for recurrence or metastasis, number of metastases, and Nottingham prognostic index had significant impact on survival. The median survival of patients determined as corresponding to Nottingham low-risk group, intermediate-risk group, and high-risk group was 29.3, 17.9, and 4.6 months, respectively. In our multivariate analysis, Karnofsky performance status (p = 0.030) and Nottingham prognostic index (p <= 0.0001) were significant prognostic factors for survival, while first-/second-/third-line chemotherapy for recurrence or metastasis (p = 0.002) was a significant predictor for the outcome of the treatment. CONCLUSIONS: The prognosis of patients with metastatic breast cancer is poor. In spite of the fact that many advances in treatment have been made, numerous additional questions have arisen; new drugs and therapeutic regimens are needed to improve the outcomes of patients, and further well-designed randomized trials are warranted. PMID- 19904564 TI - Protein recognition in ferredoxin-P450 electron transfer in the class I CYP199A2 system from Rhodopseudomonas palustris. AB - CYP199A2 from Rhodopseudomonas palustris CGA009 is a heme monooxygenase that catalyzes the oxidation of para-substituted benzoic acids. CYP199A2 activity is reconstituted by a class I electron transfer chain consisting of the associated [2Fe-2S] ferredoxin palustrisredoxin (Pux) and a flavoprotein palustrisredoxin reductase (PuR). Another [2Fe-2S] ferredoxin, palustrisredoxin B (PuxB; RPA3956) has been identified in the genome. PuxB shares sequence identity and motifs with vertebrate-type ferredoxins involved in Fe-S cluster assembly but also 50% identity with Pux and it mediates electron transfer from PuR to CYP199A2, albeit with lower steady-state turnover activity: 99 nmol (nmol P450)(-1)min(-1) for 4 methoxybenzoic acid oxidation compared with 1,438 nmol (nmol P450)(-1 )min(-1) for Pux. This difference mainly arises from weak CYP199A2-PuxB binding (K (m) 34.3 vs. 0.45 microM for Pux) rather than slow electron transfer (k (cat) 19.1 vs. 37.9 s(-1) for Pux). Comparison of the 2.0-A-resolution crystal structure of the PuxB A105R mutant with other vertebrate-type, P450-associated ferredoxins revealed similar protein folds but also significant differences in some loop regions. Therefore, PuxB offers a platform for studying ferredoxin-P450 recognition in class I P450 systems. Substitution of PuxB residues at key locations with those in Pux shows that Ala42, Cys43, and Ala44 in the [2Fe-2S] cluster binding loop and Met66 are important in electron transfer from PuxB to CYP199A2, whereas Phe73 and the C-terminal Ala105 were involved in both protein binding and electron transfer. PMID- 19904565 TI - DFT and MP2 study on the electrophilic addition reaction of bromine to exo tricyclo[3.2.1.0(2.4)]oct-6-ene. AB - The geometry and electronic structure of exo-tricyclo[3.2.1.0(2,4)]oct-6-ene (exo TCO) was investigated using DFT methods. The two faces of the endo-pyramidalised double bond of the molecule are not equivalent. The exo face of the double bond has regions with high electron density (q (i,HOMO)) and greater negative potential. Molecular complexes of exo-TCO with bromine were investigated using the B3LYP/6-311++G(d,p) method; the exo-TCO . . . Br(2)(exo) molecular complex was found to be relatively more stable than the exo-TCO . . . Br(2)(endo) complex. The cationic intermediates of the reaction were studied by DFT and MP2 methods. The solvent effect was evaluated using the self-consistent isodensity polarised continuum model (SCI-PCM). The exo-bromonium cation was found to be more stable than the endo-bromonium cation. Exo-facial selectivity due to electronic and steric factors was observed upon addition of bromine to exo-TCO. The multicentre nonclassical delocalised bromocarbonium cation IV and the exo bridged-bromonium cation I are more stable than the rearrangement cation V. The reaction products are formed via exo-bridged-bromonium I and nonclassical IV cations, which are the most stable intermediates and whose stabilities barely differ. The mechanism of the addition reaction is also discussed. PMID- 19904566 TI - High-yield fermentative preparation of tetramethylpyrazine by Bacillus sp. using an endogenous precursor approach. AB - A spore-forming Bacillus sp. was isolated from a high-temperature Daqu, a starter culture of Chinese Maotai-flavor liquor, using an endogenous precursor screening strategy. The Bacillus sp. was capable of producing a high level of 2,3,5,6 tetramethylpyrazine (TTMP) via a precursor of 3-hydroxy-2-butanone (HB). The strain was characterized as Bacillus subtilis based on morphological, physiological, and biochemical properties as well as on partial 16S rRNA gene sequences. Different carbon and nitrogen sources as well as fermentation conditions were investigated. Optimization tests showed that oxygen supply and fermentation temperature were the most important parameters determining the production process. The production of >4.08 g/l TTMP was achieved together with a high level of endogenous precursor HB accumulation (>20 g/l) in both flask and fermentor cultures when the optimized medium and cultivation conditions were applied. Our data demonstrates the effectiveness of the endogenous precursor strategy for screening microorganisms that produce flavor compounds with structure-related precursors. The high yield of TTMP and the inexpensiveness of the agro-industrial product used as the substrate (soybean meal) indicate the potential of this process for industrial application. PMID- 19904567 TI - Plant stanol supplementation decreases serum triacylglycerols in subjects with overt hypertriglyceridemia. AB - Evidence is accumulating that high serum concentrations of triacylglycerols (TAG) are, like LDL cholesterol, causally related to cardiovascular disease. A recent meta-analysis has indicated that plant stanol ester (PSE) intake not only lowered LDL cholesterol, but also serum TAG concentrations, especially in subjects with high baseline TAG concentrations. We therefore evaluated the effects of PSE supplementation on lipid metabolism in a population with elevated fasting TAG concentrations. In a randomized, placebo-controlled, parallel study, 28 subjects with elevated TAG concentrations (>1.7 mmol/L) were studied. After a 1-week run in period during which a control margarine was used, subjects consumed for 3 weeks either control or PSE-enriched margarine (2.5 g/day of plant stanols). Serum plant stanol concentrations increased in all subjects receiving the PSE enriched margarines, demonstrating good compliance. PSE supplementation significantly decreased serum total (6.7%, P = 0.015) and LDL cholesterol (9.5%, P = 0.041). A significant interaction between baseline TAG concentrations and PSE intake was found; PSE intake lowered TAG concentrations, particularly in subjects with high baseline TAG concentrations (>2.3 mmol/L; P = 0.009). Additionally, a significant interaction between baseline total number of LDL particles (LDL-P) and PSE intake was found (P = 0.020). PSE consumption lowered LDL-P, primarily in subjects with elevated baseline values; this was mainly due to a non-significant decrease in the number of atherogenic small LDL-P. Circulating levels of hs-CRP, glucose, and insulin were not changed after PSE intake. Taken together, PSE supplementation not only lowered LDL cholesterol, but also serum TAG concentrations, especially in subjects with overt hypertriglyceridemia. PMID- 19904568 TI - Heavy metal concentrations in food chain of Lake Velenjsko jezero, Slovenia: an artificial lake from mining. AB - The concentration of metals (Pb, Cd, Zn, Hg) in different ecosystem components (lake water, sediment, plankton, macrophytes, and fish tissues) has been determined in Velenjsko jezero, an artificial lake resulting from mining activity. The risk to humans from consuming fish has been evaluated from the heavy metal load of fish muscle tissue. Heavy metals are transferred through the food chain at different levels, and Hg is found to be the element of the highest ecotoxicological concern. Although both sediment and plankton contain relatively low concentrations of Hg, this element accumulates in high levels in fish, especially in the benthivorous species Abramis brama danubii and predator species Perca fluviatilis. Moreover, Hg appears to be very mobile in the fish organism. Whereas the other metals remained mostly in liver (Cd) or gills (Zn, Pb), levels of Hg in fish muscle and liver were the same and markedly higher than in gills. However, in muscle, the average concentrations of each metal were below their maximum limits, determined either by Slovenian legislation or by the Food and Agriculture Organization. PMID- 19904569 TI - East Tibetan lakes harbour novel clusters of picocyanobacteria as inferred from the 16S-23S rRNA internal transcribed spacer sequences. AB - Planktonic picocyanobacteria abundance and diversity were investigated in nine lakes on the East Tibetan Plateau spanning a salinity gradient of 0.4-22.6 g l( 1). The investigation was conducted using epifluorescence microscopy (EFM) and terminal restriction fragment polymorphism analysis of 16S-23S rRNA internal transcribed spacer (ITS) PCR amplicons followed by sequence analyses of large ITS clone libraries of seven selected samples. EFM showed that picocyanobacteria comprised 7-19% of the total prokaryotic cells found in surface water. Most of the clones were classified into six clusters and grouped within the "picocyanobacterial clade", which consists exclusively of freshwater Synechococcus. Four new phylogenetic clusters and one new subcluster of Synechococcus spp. were found, none of which are members of any known picocyanobacterial clusters. The new clusters and subcluster were the most abundant picocyanobacteria (about 96% of the sequences) in the samples collected. Sequence analyses indicated that members of the four new Synechococcus groups were only found in freshwater lakes (<1.0 g l(-1) of total dissolved solid), while members of the new subcluster were found in all the investigated Tibetan lakes, over a large salinity gradient of 0.4-22.6 g l( -1). This suggests that there is ecologically significant microdiversity within the observed Synechococcus group as defined by ITS sequences. Collectively our study demonstrated abundant and potentially novel Synechococcus in East Tibetan lakes that are likely the result of evolutionary adaptations to regional conditions. PMID- 19904570 TI - Swainsoninine concentrations and endophyte amounts of Undifilum oxytropis in different plant parts of Oxytropis sericea. AB - Locoweeds are Astragalus and Oxytropis species that contain the toxic alkaloid swainsonine. Swainsonine accumulates in all parts of the plant with the highest concentrations found in the above ground parts. A fungal endophyte, Undifilum oxytropis, found in locoweed plant species, is responsible for the synthesis of swainsonine. By using quantitative PCR, the endophyte can be quantified in locoweed species. Endophyte amounts differ between plant parts and in some instances do not mirror the concentrations of swainsonine in the corresponding parts. Two groups of Oxytropis sericea were identified: one that accumulated high concentrations of swainsonine and another where swainsonine was not detected, or concentrations were near the detection threshold. The plants with high swainsonine concentrations had quantitatively higher amounts of endophyte. Alternatively, plants with low or no swainsonine detected had quantitatively lower endophyte amounts. In addition, swainsonine and endophyte concentrations were not distributed uniformly within the same plant when separated into stalks (leaves, scape(s), and flowers/pods). These findings provide evidence as to why plants in the same population accumulate different concentrations of swainsonine, and they have important implications for sampling of locoweed plants. PMID- 19904572 TI - Postoperative morbidity and mortality of head and neck cancers in patients with liver cirrhosis undergoing surgical resection followed by microsurgical free tissue transfer. AB - BACKGROUND: The aim of this study was to evaluate the association and the related risk factors between postoperative complications and mortality and the severity of liver cirrhosis in head and neck cancer patients undergoing tumor ablation followed by microsurgical free tissue transfer. METHODS: Between January 2000 and December 2008, a total of 3108 patients were retrospectively reviewed. The diagnosis of liver cirrhosis was made mainly by abdominal ultrasonography. The Child's classification was used to assess the severity of liver cirrhosis. RESULTS: There were 60 men and 2 women enrolled. Preoperatively, 42, 17, and 3 patients were classified as Child's class A, B, and C, respectively. Class B patients had statistically significantly prolonged stay in the intensive care unit and hospital stay compared to class A patients. Patients with class B or C cirrhosis had more complications than those with class A cirrhosis (80% vs. 19.1%, P < .001). This included significantly increased rates of pulmonary complications, acute renal failure, and sepsis. The mortality rate was also significantly higher for patients with class B/C cirrhosis than for those with class A cirrhosis (30% vs. 4.8%, P = .011). By logistic regression model, preoperative platelet count, intraoperative blood transfusion > or =2 units, and Child's class were found to be significant predictive factors for morbidities. Likewise, Child's class, albumin level, intraoperative blood transfusion > or =2 units, intraoperative blood loss >500 ml, and prothrombin time were significant predictive factors for mortality. CONCLUSIONS: Child's class, along with its several components, and intraoperative blood transfusion of > or =2 units are predictive factors for morbidity and mortality. PMID- 19904573 TI - Recurrence after curative resection of early gastric cancer. AB - BACKGROUND: Few studies have evaluated the recurrence of EGC after curative gastrectomy, due to its relatively low incidence. This study evaluated recurrence patterns and independent predictive factors for recurrence in order to determine appropriate follow-up and early detection of recurrence. METHODS: We reviewed the medical records of 3,883 consecutive patients who underwent curative gastrectomy for EGC at Samsung Medical Center between February 1995 and January 2006 and were followed up until January 2008. The clinical and pathological characteristics and the predictive factors for recurrence were evaluated retrospectively. RESULTS: Eighty-five (2.2%) patients had recurrence, and liver was the most common site of recurrence (45.9%). The recurrence rates within 2, 3, and 5 years were 43.5%, 67.1%, and 85.6%, respectively. There were 106 overall recurrences, with 86 (81.1%) being detected by computed tomography (CT). Second primary cancer was the primary cause of death after gastrectomy, followed by recurrence. Old age (>60 years), tumor size (>3 cm), multiple tumor, N category, and N2 station metastasis were significant factors for recurrence by multivariate analysis, but depth of invasion was not a significant factor. CONCLUSIONS: Age, tumor size, number of tumors, N category, and N2 station metastasis were predictive factors for recurrence, with lymph node metastasis being the most significant factor. After curative gastrectomy, follow-up programs should be applied for more than 5 years for EGC patients, and computed tomography could be an essential diagnostic tool. PMID- 19904571 TI - Low temperature dormancy affects the quantity and quality of the female sexual attractiveness pheromone in red-sided garter snakes. AB - Low temperature dormancy is a necessary requirement of the annual cycle of most nonmigratory, temperate vertebrates. The red-sided garter snake, Thamnophis sirtalis parietalis, overwinters in communal dens during its prolonged winter dormancy (8 mo), and upon emergence, reproductive behavior of both sexes is maximal. Previous work on this species showed that male courtship behavior is maximally induced after simulated low temperature dormancy. The purpose of this study was to determine whether low temperature dormancy affects the pheromone profiles of individual female red-sided garter snakes. We collected females in the fall at den sites in Manitoba, Canada, and extracted pheromones from individuals at three different time points: fall (field), winter (lab), and spring (lab). Total skin lipid and pheromone fraction masses increased from fall to spring, and pheromone profiles were distinctly different in the fall and spring. Pheromone profiles became dominated by the long-chain, unsaturated methyl ketone components of the blend by the time snakes emerged in the spring. Further, the amounts of both saturated and unsaturated components increased from fall to spring, suggesting significant sex pheromone synthesis was induced by low temperature dormancy. PMID- 19904574 TI - Crushed dreams. PMID- 19904575 TI - Fruit for thought. PMID- 19904576 TI - Short-term outcomes of Roux-en-Y stapled anastomosis after distal gastrectomy for gastric adenocarcinoma. AB - BACKGROUND: Since 2003, we have begun to perform gastrojejunostomy by mechanical stapling for Roux-en-Y reconstruction in distal gastrectomy. We performed a retrospective study to compare the short-term outcomes of anastomosis by mechanical stapling and hand suturing. METHODS: We evaluated the data of 701 consecutive patients of gastric adenocarcinoma who underwent conventional open distal gastrectomy with Roux-en-Y reconstruction. The data collected included details on the method used for the Roux-en-Y reconstruction, the disease stage, extent of lymph node dissection, performance rate of truncal vagotomy, operation time, operative blood loss, length of hospital stay, and postoperative complications. RESULTS: The operation time was significantly shorter in the group in which mechanical stapling was used for the anastomosis (MS group) than in the group in which anastomosis was performed by hand suturing (HS group; 241.1 +/- 56.8 vs. 166.4 +/- 48.3 min; p < 0.05). Postoperatively, delayed gastric emptying occurred in 14 (1.9%) patients, including seven (4.2%) from the MS group and seven (1.3%) from the HS group (p = 0.038). CONCLUSION: There were no significant disadvantages of employing mechanical stapling for anastomosis, except for the high rate of delayed gastric emptying. More consideration therefore needs to be given to decreasing the frequency of gastric emptying disturbance post surgery using mechanical staples. PMID- 19904578 TI - [Severe hyponatremia associated with transcervical resection of a uterine myoma]. PMID- 19904577 TI - Recent developments in the understanding of astrocyte function in the cerebellum in vivo. AB - Several studies have contributed to our understanding of astrocytes, especially Bergmann glia, in the cerebellum; but, until recently, none has looked at their function in vivo. Multicell bolus loading of fluorescent calcium indicators in combination with the astrocytic marker SR101 has allowed imaging of up to hundreds of astrocytes at once in the intact cerebellum. In addition, the selective targeting of astrocytes with fluorescent calcium indicator proteins has enabled the study of their function in vivo without the confounding effects of other neuropil signals and with a resolution that surpasses multicell bolus loading and SR101 staining. The two astrocyte types of the cerebellar cortex, Bergmann glia, and velate protoplasmic astrocytes display a diverse signaling repertoire in vivo, which ranges from localized calcium elevations in subcellular processes to waves, triggered by the release of purines and mediated by purinergic receptors that span multiple processes and can involve tens of astrocytes. During locomotor behavior, even larger numbers of astrocytes display calcium increases that are driven by neuronal activity and correlate with global changes in blood flow. In this review, we give an overview of our current understanding of the function of Bergmann glia and velate protoplasmic astrocytes and the promise of the tools used to study their calcium dynamics and function in vivo. PMID- 19904579 TI - Spinal anesthesia for Cesarean delivery following pre-existing epidural labour analgesia. PMID- 19904580 TI - Stem cell plasticity, osteogenic differentiation and the third dimension. AB - Different cues present in the cellular environment control basic biological processes. A previously established 3D microwell array was used to study dimensionality-related effects on osteogenic differentiation and plasticity of marrow stromal cells. To enable long-term culture of single cells in the array a novel surface functionalization technique was developed, using the principle of subtractive micro contact printing of fibronectin and surface passivation with a triblock-copolymer. Immunohistochemical stainings showed that when cultivated in 3D microenvironments, marrow stromal cells can be maintained in the wells for up to 7 days and be induced to commit to the osteogenic lineage. In conclusion, this work shows the modification of a 3D microwell array allowing the long term study of single stem cell plasticity and fate in controlled microenvironments. PMID- 19904581 TI - Mathematical model to predict skin concentration of drugs: toward utilization of silicone membrane to predict skin concentration of drugs as an animal testing alternative. AB - PURPOSE: To calculate the skin concentration of active ingredients in cosmetics and topical pharmaceuticals using silicone membrane permeation. METHODS: A series of parabens were used as model ingredients. Skin concentration of parabens was calculated using silicone membrane permeability. Their partition coefficient from formulations to the silicone membrane was determined by the membrane permeation profiles, and used to calculate their silicone membrane concentration, under an assumption that the membrane is one homogenous diffusion layer. The same procedure was applied for hairless rat skin. RESULTS: The calculated concentration of parabens in silicone membrane was very close to their observed values. However, the skin concentration calculated by skin permeability was not similar to the observed concentration. Re-calculation was performed under the assumption that the skin consists of two diffusion layers. This modification using permeation data through full-thickness and stripped skin enabled precise prediction of the skin concentration of parabens. In addition, the partition coefficient to the silicone membrane was useful to estimate their skin concentration. CONCLUSIONS: Ingredient concentration in skin can be precisely predicted using diffusion equations and partition coefficients through permeation experiments using a silicone membrane. The calculated in-skin concentration is useful for formulation studies of cosmetics and topical pharmaceuticals. PMID- 19904582 TI - The targeting behavior of folate-nanohydrogel evaluated by near infrared imaging system in tumor-bearing mouse model. AB - PURPOSE: To synthesize P[(Folate-Allylamine)-co-(N-isopropylacrylamine)- co Acrylamide] (P(FoAAn-co-NIPA-AAm), folate-NHG) with appropriate diameter and lower critical solution temperature (LCST) for targeting to folate receptor (FR) expressing tumors. METHODS: Folate-NHG was synthesized by free-radical precipitation polymerization method reported in our previous work and other reports. LCST, diameter and morphology of folate-NHG were characterized by UV-vis spectrophotometer, laser particle size analyzer (LPSA) and transmission electron microscope (TEM), respectively. No.12 near infrared dye (NIRD-12) was entrapped into folate-NHG by hydrophobic association to trace the in vivo dynamic behavior of folate-NHG. This process was evaluated by a homemade near infrared (NIR) imaging system. RESULTS: Spherical folate-NHG with diameter of about 50 nm and LCST of about 40 degrees C was successfully synthesized. The photo stability of NIRD-12 was strengthened after being entrapped into folate-NHG, which enabled NIRD-12 to better trace the in vivo dynamic process of folate-NHG. Folate-NHG showed good targeting capability for all three folate receptor expressing tumor models (SMMC-7721, Bel-7402 and HeLa) with different sizes, and this accumulation could last for more than 96 h. D-folate-NHG, synthesized with double amount of FoAAn, showed better targeting effect for SMMC-7721 tumor model than that of folate-NHG. CONCLUSIONS: Folate-NHG could actively accumulate in three models of folate receptor positive tumors with different sizes and keep retention for more than 96 h, which enables it to be used as a diagnostic reagent or anti-tumor drug carrier for tumor therapy. PMID- 19904583 TI - Joint modeling of dizziness, drowsiness, and dropout associated with pregabalin and placebo treatment of generalized anxiety disorder. AB - Dizziness and drowsiness are cited as being predictors of dropout from clinical trials for the medicine pregabalin. These adverse events are typically recorded daily on a four point ordinal scale (0 = none, 1 = mild, 2 = moderate, 3 = severe), with most subjects never reporting either adverse event. We modeled the dizziness, drowsiness, and dropout associated with pregabalin use in generalized anxiety disorder using piecewise Weibull distributions for the time to first non zero dizziness or drowsiness score, after which the dizziness or drowsiness was modeled with ordinal regression with a Markovian element. Dropout was modeled with a Weibull distribution. Platykurtosis was encountered in the estimated random effects distributions for the ordinal regression models and was addressed with dynamic John-Draper transformations. The only identified predictor for the time to first non-zero dizziness or drowsiness score was daily titrated dose. Predictors for dropout included creatinine clearance and maximum daily adverse event score. Tolerance to adverse events over time was modeled by including a non stationary component for the dizziness ordinal Markov regression while the piecewise Weibull distributions allowed a change point in the median time to first non-zero dizziness or drowsiness score. PMID- 19904584 TI - Pharmacokinetic role of protein binding of mycophenolic acid and its glucuronide metabolite in renal transplant recipients. AB - Mycophenolic acid (MPA), the active compound of mycophenolate mofetil (MMF), is used to prevent graft rejection in renal transplant recipients. MPA is glucuronidated to the metabolite MPAG, which exhibits enterohepatic recirculation (EHC). MPA binds for 97% and MPAG binds for 82% to plasma proteins. Low plasma albumin concentrations, impaired renal function and coadministration of cyclosporine have been reported to be associated with increased clearance of MPA. The aim of the study was to develop a population pharmacokinetic model describing the relationship between MMF dose and total MPA (tMPA), unbound MPA (fMPA), total MPAG (tMPAG) and unbound MPAG (fMPAG). In this model the correlation between pharmacokinetic parameters and renal function, plasma albumin concentrations and cotreatment with cyclosporine was quantified. tMPA, fMPA, tMPAG and fMPAG concentration-time profiles of renal transplant recipients cotreated with cyclosporine (n = 48) and tacrolimus (n = 45) were analyzed using NONMEM. A 2- and 1-compartment model were used to describe the pharmacokinetics of fMPA and fMPAG. The central compartments of fMPA and fMPAG were connected with an albumin compartment allowing competitive binding (bMPA and bMPAG). tMPA and tMPAG were modeled as the sum of the bound and unbound concentrations. EHC was modeled by transport of fMPAG to a separate gallbladder compartment. This transport was decreased in case of cyclosporine cotreatment (P < 0.001). In the model, clearance of fMPAG decreased when creatinine clearance (CrCL) was reduced (P < 0.001), and albumin concentration was correlated with the maximum number of binding sites available for MPA and MPAG (P < 0.001). In patients with impaired renal function cotreated with cyclosporine the model adequately described that increasing fMPAG concentrations decreased tMPA AUC due to displacement of MPA from its binding sites. The accumulated MPAG could also be reconverted to MPA by the EHC, which caused increased tMPA AUC in patients cotreated with tacrolimus. Changes in CrCL had hardly any effect on fMPA exposure. A decrease in plasma albumin concentration from 0.6 to 0.4 mmol/l resulted in ca. 38% reduction of tMPA AUC, whereas no reduction in fMPA AUC was seen. In conclusion, a pharmacokinetic model has been developed which describes the relationship between dose and both total and free MPA exposure. The model adequately describes the influence of renal function, plasma albumin and cyclosporine co-medication on MPA exposure. Changes in protein binding due to altered renal function or plasma albumin concentrations influence tMPA exposure, whereas fMPA exposure is hardly affected. PMID- 19904585 TI - D-optimal designs for parameter estimation for indirect pharmacodynamic response models. AB - This report generates efficient experimental designs (dose, sampling times) for parameter estimation for four basic physiologic indirect pharmacodynamic response (IDR) models. The principles underlying IDR models and their response patterns have been well described. Each IDR model explicitly contains four parameters, k (in) (production), k (out) (loss), I (max)/S (max) (capacity) and IC (50)/SC (50) (sensitivity). The pharmacokinetics of an IV dose of drug described by a monoexponential function of time with two parameters, V and k (el), is assumed. The random errors in the response variable are assumed to be additive, independent, and normal with zero mean and variance proportional to some power of the mean response. Optimal design theory was used extensively to assess the role of both dose and sampling times. Our designs were generated in Mathematica (ADAPT 5 typically produces identical results). G-optimality was used to verify that the generated designs were indeed D-optimal. Such designs are efficient and robust when good prior knowledge of the estimated parameters is available. The efficiency of unconstrained D-optimal designs (4 dose, sampling time pairs) does not improve much when the drug doses are allowed to differ, compared with constrained single dose designs (4 sampling times) with one maximal feasible dose. Also, explored were efficiencies of alternative study designs and results from parameter misspecification. This analysis substantiates the importance of larger doses yielding greater certainty in parameter estimation in pharmacodynamics. PMID- 19904587 TI - Examining the relationship between genetic counselors' attitudes toward deaf people and the genetic counseling session. AB - Given the medical and cultural perspectives on deafness it is important to determine if genetic counselors' attitudes toward deaf people can affect counseling sessions for deafness genes. One hundred fifty-eight genetic counselors recruited through the National Society of Genetic Counselors Listserv completed an online survey assessing attitudes toward deaf people and scenario specific comfort levels discussing and offering genetic testing for deafness. Respondents with deaf/Deaf friends or who work in prenatal or pediatric settings had more positive attitudes toward deaf people than those without deaf/Deaf friends or those working in 'other' settings. More positive attitudes toward deaf people correlated with higher comfort level talking about genetic testing for the two scenarios involving culturally Deaf clients; and correlated with higher comfort level offering genetic testing to culturally Deaf clients wishing to have a deaf child. Attitudes and comfort level were not correlated in the scenarios involving hearing or non-culturally deaf clients. These results suggest that genetic counselors' attitudes could affect information provision and the decision making process of culturally Deaf clients. Cultural sensitivity workshops in genetic counseling training programs that incorporate personal interactions with culturally Deaf individuals are recommended. Additional suggestions for fostering personal interactions are provided. PMID- 19904586 TI - Clinical characteristics and genotype-phenotype correlation in 62 patients with X linked agammaglobulinemia. AB - INTRODUCTION: X-linked agammagobulinemia (XLA) is a primary immunodeficiency disorder caused by Bruton's tyrosine kinase (Btk) gene mutation. Recent studies suggested genotype-phenotype correlation in XLA, but a definitive association remains controversial. PATIENTS AND METHODS: We examined the relationship between specific Btk gene mutations and severity of clinical presentation in 62 patients with XLA. Disease severity was assessed by the age of disease onset and the presence of severe infections, while mutations were classified into severe and mild based on structural and functional consequence by bioinformatics analysis. RESULTS: Fifty-six Btk mutations were identified in 62 patients from 57 kindreds. Variation in phenotypes was observed, and there was a tendency of association between genotype and age of disease onset as well as occurrence of severe infections. CONCLUSION: A critical analysis of the circumstances upon presentation also revealed that under-recognition of recurrent infections and relevant family history are important hurdles to timely diagnosis of XLA. PMID- 19904588 TI - Mitochondrial respiratory dysfunction and mutations in mitochondrial DNA in PINK1 familial parkinsonism. AB - A summary is presented of the cellular function and topology of the protein products of genes whose mutations are associated with familial forms of parkinsonism, with particular emphasis on mitochondrial involvement. Observations are reviewed which show mitochondrial respiratory depression in the fibroblasts of a patient affected by familial parkinsonism associated with homozygous PINK1 mutation. The respiratory depression, which was due to loss of mitochondrial cytochrome c, was associated with decreased capacity of respiratory chain oxidative phosphorylation and enhanced cellular level of ROS. Sequence analysis of the overall mtDNA revealed coexistence with the PINK1 mutation of homoplasmic point mutations in the ND5 and ND6 genes of complex I. The presence of these mutations appears to have an impact on the development of the parkinsonism, which can also occur in the heterozygous PINK1 mutation state. PMID- 19904589 TI - PET studies of cerebral metabolism in Parkinson disease. AB - A defect in cerebral energy production due to dysfunction of the mitochondrial electron transport system (ETS) has been postulated to be important in the pathogenesis of Parkinson Disease (PD). However, direct in vivo measurements of cerebral mitochondrial function are scant and inconsistent. We directly investigated cerebral mitochondrial function in vivo with positron emission tomography (PET) in 12 patients with early, never-medicated PD and 12 age-matched normal controls by combined measurements of the cerebral metabolic rate of oxygen (CMRO(2)) and the cerebral metabolic rate of glucose (CMRglc). Instead of the decrease in CMRO(2) and CMRO(2)/CMRglc molar ratio characteristic of defects in mitochondrial oxidative metabolism, there was a statistically significant 24% general increase in CMRO(2) and no change in CMRO(2)/CMRglc. Since PD symptoms were already manifest, reduced oxidative activity of the mitochondrial ETS cannot be a primary mechanism of neuronal death in early PD. This increase in metabolism could reflect the increased energy requirements of an injured brain or an uncoupling of ATP production from oxidation in the terminal stage of oxidative phosphorylation. Which is the case in early PD and whether these metabolic abnormalities are important in the pathogenesis of PD will require further study. PMID- 19904594 TI - Subject of the year: who are we missing, who are we overtreating, and who is best served? Refining the prescription of implantable cardioverter-defibrillator therapy. PMID- 19904590 TI - Parkinson's disease and mitochondrial complex I: a perspective on the Ndi1 therapy. AB - Mitochondrial impairment has been collecting more and more attention as a contributing factor to the etiology of Parkinson's disease. Above all, the NADH quinone oxidoreductase, complex I, of the respiratory chain seems to be most culpable. Complex I dysfunction is translated to an increased production of reactive oxygen species and a decreased energy supply. In the brain, the dopaminergic neurons are one of the most susceptible cells. Their death is directly linked to the disease apparition. Developing an effective gene therapy is challenged by harmful actions of reactive oxygen species. To overcome this problem a therapeutic candidate must be able to restore the NADH-quinone oxidoreductase activity regardless of how complex I is impaired. Here we discuss the potency of the yeast alternative NADH dehydrogenase, the Ndi1 protein, to reinstate the mitochondrial respiratory chain compensating for disabled complex I and the benefit Ndi1 brings toward retardation of Parkinson's disease. PMID- 19904593 TI - An examination of prepartying and drinking game playing during high school and their impact on alcohol-related risk upon entrance into college. AB - Prepartying and drinking game playing are associated with excessive alcohol consumption and alcohol-related negative consequences in college populations; however, research exploring the prevalence of these high risk drinking contexts among high school students, and how such engagement may impact both high school and subsequent college drinking risk, is lacking. The current study, which is the first study to assess prepartying during high school, examined how engaging in either prepartying or drinking game playing during high school was associated with risky high school drinking as well as alcohol use and consequences during the transitional first month of college. The study involved 477 first-year college students, the majority of whom were 18 years old (94%), female (66%), and Caucasian (59%). Prepartying was found to be highly prevalent in high school (45%). Further, students who prepartied or played drinking games during high school drank significantly more in high school than students who did not engage in these high risk activities. Finally, prepartying and game playing during high school were associated with greater collegiate alcohol consumption (controlling for high school drinking) and consequences (controlling for both high school and college drinking). This study establishes prepartying and drinking games as common high risk activities among both high school and incoming first-year college students, and addresses implications for prevention and targeted interventions. PMID- 19904595 TI - Comparison of brain-computer interface decoding algorithms in open-loop and closed-loop control. AB - Neuroprosthetic devices such as a computer cursor can be controlled by the activity of cortical neurons when an appropriate algorithm is used to decode motor intention. Algorithms which have been proposed for this purpose range from the simple population vector algorithm (PVA) and optimal linear estimator (OLE) to various versions of Bayesian decoders. Although Bayesian decoders typically provide the most accurate off-line reconstructions, it is not known which model assumptions in these algorithms are critical for improving decoding performance. Furthermore, it is not necessarily true that improvements (or deficits) in off line reconstruction will translate into improvements (or deficits) in on-line control, as the subject might compensate for the specifics of the decoder in use at the time. Here we show that by comparing the performance of nine decoders, assumptions about uniformly distributed preferred directions and the way the cursor trajectories are smoothed have the most impact on decoder performance in off-line reconstruction, while assumptions about tuning curve linearity and spike count variance play relatively minor roles. In on-line control, subjects compensate for directional biases caused by non-uniformly distributed preferred directions, leaving cursor smoothing differences as the largest single algorithmic difference driving decoder performance. PMID- 19904596 TI - Brief Report: Randomized test of the efficacy of picture exchange communication system on highly generalized picture exchanges in children with ASD. AB - A randomized control trial comparing two social-communication interventions in young children with autism examined far-transfer of the use of picture exchange to communicate. Thirty-six children were randomly assigned to one of two treatment conditions, one of which was the Picture Exchange Communication System (PECS). All children had access to picture symbols during assessments. Post treatment measurement of the number of picture exchanges in a far-transfer, assessment context favored the PECS intervention. These findings were interpreted as support for the hypothesis that the PECS curriculum can successfully teach a generalized means of showing coordinated attention to object and person without requiring eye contact to children with ASD. PMID- 19904597 TI - Do gaze cues in complex scenes capture and direct the attention of high functioning adolescents with ASD? Evidence from eye-tracking. AB - Visual fixation patterns whilst viewing complex photographic scenes containing one person were studied in 24 high-functioning adolescents with Autism Spectrum Disorders (ASD) and 24 matched typically developing adolescents. Over two different scene presentation durations both groups spent a large, strikingly similar proportion of their viewing time fixating the person's face. However, time-course analyses revealed differences between groups in priorities of attention to the region of the face containing the eyes. It was also noted that although individuals with ASD were rapidly cued by the gaze direction of the person in the scene, this was not followed by an immediate increase in total fixation duration at the location of gaze, which was the case for typically developing individuals. PMID- 19904598 TI - Parents' views and experiences about complementary and alternative medicine treatments for their children with autistic spectrum disorder. AB - Use of complementary and alternative medicine (CAM) treatments have been increasing for children with autistic spectrum disorder (ASD). In this study, 38 Turkish parents of children with ASD were surveyed related with their use of CAM treatments, experiences, and views for each treatment. They mentioned "Vitamins and minerals", "Special Diet", "Sensory Integration", "Other Dietary Supplements", and "Chelation" as five frequently used CAM treatments. Communication, learning, health, and behavior were the main four areas rated as "improved" after five CAM treatments. Negative sides of treatments were listed as being expensive, difficult to apply, or harmful. The parents' views on some treatments have varied from great improvement to worse. Reported improvements were considerably higher than the negative sides of the treatments. PMID- 19904599 TI - HIV in the leather community: rates and risk-related behaviors. AB - There exist many subcultures of men who have sex with men (MSM), all with differing values and health behaviors. The Leathermen comprise one such subculture, which is characterized by a heightened valuation of hypersexuality and adherence to sexual control dynamics (i.e., submission and dominance). No previous research has specifically examined this community for differences in sexual health (e.g., HIV rates) and sexual health behaviors (e.g., condom use). We conducted a large survey of men (N = 1,554) at one leather and non-leather event, collecting data from 655 Submissives, Dominants, Switches, and non orienting Leathermen. Leathermen were 61% more likely to be HIV-positive than non Leathermen. Decreased condom use found in HIV-positive Leathermen (relative to HIV-positive non-Leathermen) was a potential factor contributing to heightened HIV rates. Universal low condom use in Submissives engaging in receptive, and Dominants engaging in insertive, anal intercourse was an additional trend that potentially contributed to increased numbers of HIV-positive Leathermen. Our recommendation is for heightened awareness of the risks associated with sex among Leathermen, especially unprotected anal intercourse with sero-uncertain Submissives. PMID- 19904600 TI - Vitrification of bovine oocytes: implications of follicular size and sire on the rates of embryonic development. AB - PURPOSE: The objectives were to test how the source of oocytes and semen impacted vitrification of large numbers of bovine oocytes and subsequent IVF and early embryo development to test procedures that may assist with assisted reproductive technologies in humans. METHODS: Bovine oocytes were vitrified from follicles of different diameters, small (< or =4 mm) and medium (4 to 10 mm), using nylon mesh. Oocytes were exposed to the cryoprotectant composed of 40% (v/v) ethylene glycol, 18% (w/v) Ficoll-70, and 0.3 M sucrose in three stepwise dilutions. Thawing was conducted with a series of 0.5, 0.25 and 0.125 M sucrose dilutions in 20% fetal bovine serum. RESULTS: The cleavage (39.1% vs. 58.5%) and blastocyst rates (5.1% vs. 22.9%) were significantly lower for the vitrified oocytes. Follicle size had a significant impact on the development of embryos. Sires had significant effects on embryonic developmental rates. CONCLUSIONS: We conclude that differences in development exist due to follicle source and sire used for IVF after vitrification. PMID- 19904601 TI - An auto-controlled prospective comparison of two embryos culture media (G III series versus ISM) for IVF and ICSI treatments. AB - PURPOSE: To compare the effects of 2 different media on embryo morphology and development at days 2/3. METHOD: Six hundred seventy-six attempts from 512 couples were included in this prospective auto-controlled study. Sibling oocytes of all couples undergoing an IVF (n = 286) or ICSI (n = 390) attempt were randomly assigned to either GIII series (Vitrolife) or ISM (Medicult) media. Primary end points were fertilization and embryo morphology rates. RESULTS: Fertilization rates in GIII series and ISM (IVF: 59.9 vs 62.0% and ICSI: 65.7 vs 66.8%) respectively were not different. GIII series showed an increase, compared to ISM, of early cleavage rate, (IVF: 25.8 vs 16.2% (p = 0.005); ICSI: 40.8 vs 25.5% (p < 0.0001), and good embryo morphology rate at day 2 [IVF: 64.6 vs 57.3% (p = 0.01); ICSI: 74.2 vs 69.4 (p = 0.03)] and at day 3 [IVF: 57.5 vs 49.0% (p = 0.02); ICSI: 67.2 vs 61.6% (p = 0.01)]. CONCLUSIONS: Embryo morphology at days 2/3 was significantly enhanced when the embryos were cultured in GIII series. PMID- 19904602 TI - Environmental contributions to the stability of antisocial behavior over time: are they shared or non-shared? AB - It has recently been argued that shared environmental influences are moderate, identifiable, and persistent sources of individual differences in most forms of child and adolescent psychopathology, including antisocial behavior. Unfortunately, prior studies examining the stability of shared environmental influences over time were limited by possible passive gene-environment correlations, shared informants effects, and/or common experiences of trauma. The current study sought to address each of these limitations. We examined adolescent self-reported antisocial behavior in a 3.5 year longitudinal sample of 610 biological and adoptive sibling pairs from the Sibling Interaction and Behavior Study (SIBS). Results revealed that 74-81% of shared environmental influences present at time 1 were also present at time 2, whereas most non-shared environmental influences (88-89%) were specific to a particular assessment period. Such results provide an important constructive replication of prior research, strongly suggesting that shared environmental contributions to antisocial behavior are systematic in nature. PMID- 19904603 TI - Some common mutations of RAD50 and NBS1 in western populations do not contribute significantly to Chinese non-BRCA1/2 hereditary breast cancer. PMID- 19904605 TI - Fatty acid composition of frontal, temporal and parietal neocortex in the normal human brain and in Alzheimer's disease. AB - Dietary omega3-polyunsaturated fatty acids are thought to influence the risk of Alzheimer's disease (AD), and supplemental docosahexaenoic acid (DHA; 22:6n-3) has been reported to reduce neurodegeneration in mouse models of AD. We have analysed the fatty acid composition of frontal, temporal and parietal neocortex in 58 normal and 114 AD brains. Significant reductions were found for stearic acid (18:0) in frontal and temporal cortex and arachidonic acid (20:4n-6) in temporal cortex in AD, and increases in oleic acid in frontal and temporal cortex (18:1n-9) and palmitic acid (16:0) in parietal cortex. DHA level varied more in AD than controls but the mean values were not significantly different. Fatty acid composition was not related to APOE genotype, age, gender or post-mortem delay. Further research is needed to distinguish between alterations that are secondary to AD and those that contribute to the disease process. PMID- 19904606 TI - In celebration of the life and work of Donald C. Klein. PMID- 19904607 TI - Paul Tillich and psychoanalysis. AB - Paul Tillich (1886-1965) was one of the leading theologians of the twentieth century. Tillich was born in Germany and received his education and first academic appointments there. Tillich left Germany in 1933 to teach at Union Theological Seminary after having been dismissed from his university position by the National Socialist government for his radical views and political associations. In the United States, he became a highly successful lecturer, preacher, and public intellectual who reached numbers of persons who had departed or who had doubts regarding traditional religious belief and practice. Tillich underwent a series of traumatic losses in the early decades of his life that powerfully shaped his subsequent contributions to religious and cultural discourse. This essay outlines this pattern of loss and speculates about its impact upon his theological work. It lifts up Tillich's perspective of living and working "on the boundary" of disciplines, eras, and cultures, most particularly where psychoanalytic ideas contributed to his "theology of culture." It also stresses Tillich's role in initiating the ongoing dialogue between religion and psychiatry and psychoanalysis. The essay concludes with a summary critique of Tillich's work along with an affirmation of his considerable legacy. This essay was originally a presentation for the Richardson Research Seminar in the History of Psychiatry at Weill Cornell Medical College. PMID- 19904609 TI - Serum deprivation-induced reactive oxygen species production is mediated by Romo1. AB - Serum deprivation-triggered increases in reactive oxygen species (ROS) are known to induce apoptotic cell death. However, the mechanism by which serum deprivation causes ROS production is not known. Since mitochondria are the main source of ROS and since mitochondrial ROS modulator 1 (Romo1) is involved in ROS production, we sought to determine if serum deprivation triggered ROS production through Romo1. To examine the relationship between Romo1 and the serum deprivation-triggered increase in ROS, we transfected Romo1 siRNA into various cell lines and looked for inhibition of mitochondrial ROS generation. Romo1 knockdown by Romo1 siRNA blocked the mitochondrial ROS production caused by serum deprivation, which originates in the mitochondrial electron transport chain. We also found that Romo1 knockdown inhibited serum deprivation-induced apoptosis. These findings suggest that Romo1-derived ROS play an important role in apoptotic cell death triggered by withdrawal of cell survival factors. PMID- 19904608 TI - Is religious coping associated with cumulative health risk? An examination of religious coping styles and health behavior patterns in Alzheimer's dementia caregivers. AB - The current study explored the relationship between religious coping and cumulative health risk associated with health behavior patterns in a sample of 256 Latina and Caucasian female caregivers of elderly relatives with dementia. Primary analyses examined the relationship between religious coping (both positive and negative) and an overall index of cumulative health risk. Secondary analyses were conducted on the individual health behaviors subsumed in the broader index. Findings revealed that negative religious coping was significantly associated with increased cumulative health risk. Positive religious coping was predictive of decreased cumulative health risk among Latina caregivers but not among Caucasians. Negative religious coping was significantly associated with both an increased likelihood for weight gain and increased dietary restriction. Positive religious coping was associated with decreased likelihood for weight gain in Latinas. Implications for both caregivers and clinicians are discussed. PMID- 19904610 TI - Inhibition of peptidyl-prolyl cis/trans isomerase Pin1 induces cell cycle arrest and apoptosis in vascular smooth muscle cells. AB - This study was undertaken to determine the in vitro effect of lentivirus-mediated siPin1 on cell cycle and apoptosis of vascular smooth muscle cells (VSMCs). Further we sought to provide insight into the mechanisms behind these processes. Human umbilical artery smooth muscle cells (HUASMCs) were transfected with lentiviral siPin1. Real-time RT-PCR and Western blotting were used to examine Pin1 mRNA and protein expression. MTT and [(3)H]thymidine incorporation assays were employed to observe cell proliferation status. The apoptotic rate and cell cycle were analyzed by Hoechst33258 staining and flow cytometry. Finally we measured the expression of cyclin D1, beta-catenin, CDK4, cytochrome c, procaspase-3, cleaved caspase-3, procaspase-9, cleaved caspase-9, Bcl-2, Bax, STAT3, phosphorylated STAT3 and VEGF in lentiviral siPin1 infected VSMCs. Lentivirus-mediated siPin1 effectively diminished endogenous Pin1 expression in VSMCs resulting in cell cycle arrest and enhancement of apoptosis. This was accompanied by downregulation of cyclin D1, beta-catenin, CDK4, increase of Bax/Bcl-2 ratio, release of cytochrome c, and activation of caspase-3 and -9. We concluded that this effect was mediated, at least in part, via the beta catenin/cyclin D1/CDK4 cascade, and that the mitochondrial pathway was responsible for VSMC apoptosis in the absence of Pin1. Our observations raised the possibility that Pin1 might be a potential therapeutic target to prevent stenosis. PMID- 19904612 TI - Acromegaly with negative pituitary MRI and no evidence of ectopic source: the role of transphenoidal pituitary exploration? AB - Growth hormone (GH) producing adenomas of the pituitary gland are usually macroadenomas (>10 mm in size). Often these adenomas are locally invasive by the time of diagnosis. Acromegaly secondary to a very small pituitary microadenoma not visualized on pituitary magnetic resonance (MR) imaging is rare. We report a patient with acromegaly and an unremarkable pituitary MR imaging who had negative work up for ectopic growth hormone-releasing hormone (GHRH) or GH secreting tumors. Transsphenoidal pituitary exploration revealed a pituitary adenoma located on the left side of the sella against the medial wall of the cavernous sinus extending posteriorly along the floor of the sella all the way to the right side. The acromegaly was treated with resection of the pituitary adenoma and normalization of serum insulin-like growth factor 1 (IGF-1) and GH levels. In a patient with acromegaly and unremarkable pituitary MR imaging, with no evidence of ectopic GH and GHRH production, transsphenoidal pituitary exploration is a reasonable approach and may result in clinical improvement and biochemical cure in the hand of experienced surgeon. This approach may avoid long term medical treatment with its associated cost. PMID- 19904611 TI - Antioxidant activity and total phenolic content of Moringa oleifera leaves in two stages of maturity. AB - Antioxidants play an important role in inhibiting and scavenging free radicals, thus providing protection to human against infections and degenerative diseases. Current research is now directed towards natural antioxidants originated from plants due to safe therapeutics. Moringa oleifera is used in Indian traditional medicine for a wide range of various ailments. To understand the mechanism of pharmacological actions, antioxidant properties of the Moringa oleifera leaf extracts were tested in two stages of maturity using standard in vitro models. The successive aqueous extract of Moringa oleifera exhibited strong scavenging effect on 2, 2-diphenyl-2-picryl hydrazyl (DPPH) free radical, superoxide, nitric oxide radical and inhibition of lipid per oxidation. The free radical scavenging effect of Moringa oleifera leaf extract was comparable with that of the reference antioxidants. The data obtained in the present study suggests that the extracts of Moringa oleifera both mature and tender leaves have potent antioxidant activity against free radicals, prevent oxidative damage to major biomolecules and afford significant protection against oxidative damage. PMID- 19904614 TI - Uptake of biodegradable gel-assisted LBL nanomatrix by Leishmania donovani infected macrophages. AB - The aim of this study was to develop novel gel-assisted layer-by-layer (LBL) nanomatrix with high payload of doxorubicin (DOX) and to assess its efficacy against Leishmania donovani. The biodegradable LBL nanomatrix was fabricated using LBL technique using polyions (protamine and sodium alginate) on decomposable core. The developed system was characterized in vitro in terms of layer-by-layer growth and payload efficiency. The efficacy of optimized formulations was evaluated against L. donovani strain in terms of inhibitory concentration (IC50). Uptake studies by infected macrophages were investigated both qualitatively and quantitatively using fluorescence microscopy and flow cytometry. The autogelling property subsequent to core removal inside the nanomatrix resulted in high payload efficiency of DOX (i.e., >70%). The reversal in charge followed the same trend with additional layers, and the magnitude of the charge remained constant up to five complete bilayers of polyions. The DOX can be effectively encapsulated, delivered, and subsequently taken up by L. donovani-infected macrophage cells. The matrix is completely internalized into macrophages showing improved efficacy (IC50 of formulation is almost 2500. A rank and percentile procedure resulted in a 50th percentile WPF of 270 and a lower 5th percentile estimate > 50. These WPFs exceed the assigned protection factor of 10 for half facepieces published by the Occupational Safety and Health Administration. This study's results support the APF of 10 and indicate the respirator provided appropriate protection as it was used in this study. The comparability of the two analytical methods commonly used together in WPF studies was also evaluated. The samples collected outside the respirators were analyzed for lead by proton-induced X-ray emission analysis (PIXE) followed by inductively coupled plasma spectrometry (ICP). While the two methods were highly correlated (r(2) = 0.965), the mean PIXE lead mass was approximately 45% higher than the mean ICP value. This systematic bias was explained by the assumptions used to interpret the PIXE analytical results. When WPF studies use ICP and PIXE for C(o) and C(i) samples, respectively, the calculated WPF values are conservative estimates of respirator performance. PMID- 19904660 TI - Occupational exposure to cobalt: a population toxicokinetic modeling approach validated by field results challenges the biological exposure index for urinary cobalt. AB - This study modeled the urinary toxicokinetics of cobalt exposure based on 507 urine samples from 16 workers, followed up for 1 week, and 108 related atmospheric cobalt measurements to determine an optimal urinary cobalt sampling strategy at work and a corresponding urinary exposure threshold (UET). These data have been used to calibrate a population toxicokinetic model, taking into account both the measurement uncertainty and intra- and interindividual variability. Using the calibrated model, urinary sampling sensitivity and specificity performance in detecting exposure above the 20 microg/m(3) threshold limit value time-weighted average (TLV-TWA) has been applied to identify an optimal urine sampling time. The UET value is obtained by minimizing misclassification rates in workplace exposures below or above the TLV. Total atmospheric cobalt concentrations are in the 5-144 microg/m(3) range, and total urinary cobalt concentrations are 0.5-88 microg/g creatinine. A two-compartment toxicokinetic model best described urinary elimination. Terminal elimination half-time from the central compartment is 10.0 hr (95% confidence interval [8.3-12.3]). The optimal urinary sampling time has been identified as 3 hr before the end of shift at the end of workweek. If we assume that misclassification errors are of equal cost, the UET associated with the TLV of 20 microg/m(3) is 5 microg/L, which is lower than the ACGIH-recommended biological exposure index of 15 microg/L. PMID- 19904661 TI - Diminished speech intelligibility associated with certain types of respirators worn by healthcare workers. AB - This study sought to determine the level of communication interference associated with commonly used disposable and reusable respirators and surgical masks worn by healthcare workers. Speech intelligibility was assessed using the modified rhyme test in an intensive care unit environment. Respirators decreased speech intelligibility by a range of 1% to 17%, although not all were statistically significant. Differences in speech intelligibility associated with surgical masks and disposable filtering facepiece respirators (without exhalation valves) were not statistically significant compared with controls. Wearing half-face elastomeric respirators with voice augmentation equipment was associated with higher speech intelligibility than models without this equipment (OR = 2.81). Hearing clarity while wearing a powered air-purifying respirator (PAPR) was 79% compared with 90% with no PAPR (OR = 0.40). While some respirators appear to have little or no effect on speech intelligibility, interference with speech intelligibility associated with certain types of respirators commonly worn by U.S. healthcare workers may be substantial. PMID- 19904663 TI - Mindfulness-based therapies for substance use disorders: part 1. PMID- 19904664 TI - Mindfulness meditation for substance use disorders: a systematic review. AB - Relapse is common in substance use disorders (SUDs), even among treated individuals. The goal of this article was to systematically review the existing evidence on mindfulness meditation-based interventions (MM) for SUDs. The comprehensive search for and review of literature found over 2000 abstracts and resulted in 25 eligible manuscripts (22 published, 3 unpublished: 8 randomized controlled trials, 7 controlled nonrandomized, 6 noncontrolled prospective, and 2 qualitative studies, and 1 case report). When appropriate, methodological quality, absolute risk reduction, number needed to treat, and effect size were assessed. Overall, although preliminary evidence suggests MM efficacy and safety, conclusive data for MM as a treatment of SUDs are lacking. Significant methodological limitations exist in most studies. Further, it is unclear which persons with SUDs might benefit most from MM. Future trials must be of sufficient sample size to answer a specific clinical question and should target both assessment of effect size and mechanisms of action. PMID- 19904665 TI - Mindfulness-based relapse prevention for substance use disorders: a pilot efficacy trial. AB - The current study is the first randomized-controlled trial evaluating the feasibility and initial efficacy of an 8-week outpatient Mindfulness-Based Relapse Prevention (MBRP) program as compared to treatment as usual (TAU). Participants were 168 adults with substance use disorders who had recently completed intensive inpatient or outpatient treatment. Assessments were administered pre-intervention, post-intervention, and 2 and 4 months post intervention. Feasibility of MBRP was demonstrated by consistent homework compliance, attendance, and participant satisfaction. Initial efficacy was supported by significantly lower rates of substance use in those who received MBRP as compared to those in TAU over the 4-month post-intervention period. Additionally, MBRP participants demonstrated greater decreases in craving, and increases in acceptance and acting with awareness as compared to TAU. Results from this initial trial support the feasibility and initial efficacy of MBRP as an aftercare approach for individuals who have recently completed an intensive treatment for substance use disorders. PMID- 19904666 TI - Mindfulness training and stress reactivity in substance abuse: results from a randomized, controlled stage I pilot study. AB - Stress is important in substance use disorders (SUDs). Mindfulness training (MT) has shown promise for stress-related maladies. No studies have compared MT to empirically validated treatments for SUDs. The goals of this study were to assess MT compared to cognitive behavioral therapy (CBT) in substance use and treatment acceptability, and specificity of MT compared to CBT in targeting stress reactivity. Thirty-six individuals with alcohol and/or cocaine use disorders were randomly assigned to receive group MT or CBT in an outpatient setting. Drug use was assessed weekly. After treatment, responses to personalized stress provocation were measured. Fourteen individuals completed treatment. There were no differences in treatment satisfaction or drug use between groups. The laboratory paradigm suggested reduced psychological and physiological indices of stress during provocation in MT compared to CBT. This pilot study provides evidence of the feasibility of MT in treating SUDs and suggests that MT may be efficacious in targeting stress. PMID- 19904667 TI - Associations of mindfulness with nicotine dependence, withdrawal, and agency. AB - Quitting smoking is a major life stressor that results in numerous aversive consequences, including persistently increased level of post-cessation negative affect and relapse. The identification of factors that may enhance behavioral and emotional regulation after quitting may be useful in enhancing quit rates and preventing relapse. One factor broadly linked with behavioral and emotional regulation is mindfulness. This study examined baseline associations of mindfulness with demographic variables, smoking history, dependence, withdrawal severity, and agency among 158 smokers enrolled in a cessation trial. Results indicated that mindfulness was negatively associated with level of nicotine dependence and withdrawal severity, and positively associated with a sense of agency regarding cessation. Moreover, mindfulness remained significantly associated with these measures even after controlling for key demographic variables. Results suggest that low level of mindfulness may be an important predictor of vulnerability to relapse among adult smokers preparing to quit; thus, mindfulness-based interventions may enhance cessation. PMID- 19904668 TI - Associations between mindfulness and implicit cognition and self-reported affect. AB - Theory suggests that mindful individuals exhibit enhanced attentional processing (e.g., attentional control) and that they maintain a detached perspective to problematic stimuli. For smokers, smoking and affective stimuli are problematic stimuli when they try to quit. In this cross-sectional study, smokers (n = 158) completed 3 modified Stroop tasks (to assess attentional control), 3 Implicit Association Tests (IATs; to assess detached perspective), and a battery of self report assessments. Degree of mindfulness was negatively associated (P < .05) with self-reported negative affect, perceived stress, and depressive symptom severity, and positively associated (P < .05) with positive affect. Degree of mindfulness was not associated with the ability to disengage attention from smoking or affective stimuli. On the depression IAT, more mindful participants exhibited a more negative IAT effect, suggesting that they may have developed a detached perspective to depression-related stimuli. Theoretical and clinical implications of the data are discussed. PMID- 19904671 TI - [Abstracts from conference on quality measurement and quality management with routine data]. PMID- 19904670 TI - Pushing quality in health care management. PMID- 19904697 TI - [Abstracts of the 3rd National Congress on Preventive Medicine. November 27-28. 2009. Dresden, Germany]. PMID- 19904698 TI - Osteopetrosis due to homozygous chloride channel ClCN7 mutation mimicking metabolic disease with haematological and neurological impairment. AB - We report on the fatal clinical course of a 3 year old male Turkish patient suffering from osteopetrosis caused by a homozygous mutation in the chloride channel gene ClCN7 with developing pancytopenia and severe neurological impairment. Hepatosplenomegaly due to extramedullary hematopoesis, severe transfusion-dependent anemia and growth failure initially suggested metabolic or oncologic disorder. Particular haematological parameters like tear drop cells basophilic punctation of the polymorphonuclear cells in the absence of haemolysis caused the diagnostic X-ray investigations of the skull and vertebral column. Raised serum creatinkinase-BB isoenzyme and genetic testing were in line with the diagnose of osteopetrosis at an age of 2(1/2) years. CONCLUSION: Osteopetrosis is a rare but considerable differential diagnose for unclarified change in haematopoetic cell lines combined with severe neurological symptoms mimicking metabolic or haematological disease. Because of this rare disease a consensus protocol for diagnostics, treatment and follow up of patients suffering from osteopetrosis is recently worked out from the European Group of Blood and Marrow Transplantation (EBMT) and the European Society for Immundeficiencies (ESID) to build up a central registry for this disease (available by ansgar.schulz@uniklinik-ulm.de). PMID- 19904699 TI - Ulcers and gastritis. PMID- 19904700 TI - Laser-supported diaphanoscopy: an innovative technique for locating gastric stromal tumors in gastroscopic-laparoscopic rendezvous: a case series. AB - Development of an innovative method of endoscopic laser-supported diaphanoscopy, for precise demonstration of the location of gastrointestinal stromal tumors (GISTs) at laparoscopy is described. The equipment consists of a light transmission cable with an anchoring system for the gastric mucosa, a connecting system for the light source, and the laser light source itself. During surgery, transillumination by laser is used to show the shape of the tumor. The resection margins are then marked by electric coagulation. Ten patients have been successfully treated using this technique in laparoscopic-endoscopic rendezvous procedures. Average time of surgery was 123 minutes. The time for marking the shape of the tumor averaged 16 minutes. Depending on tumor location and size, 4-7 marks were used, and resection margins were 4-15 mm. This new and effective technique facilitates precise locating of gastric GISTs leading to exact and tissue-sparing transmural laparoscopic resections. PMID- 19904701 TI - Significant clinical implications of prophylactic pancreatic stent placement in previously normal pancreatic ducts. AB - Pancreatic duct stent placement is increasingly performed for the prevention of pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP); however stents can result in injury especially in normal ducts. The clinical significance and outcomes of subsequent endoscopic therapy are unknown. This study was a retrospective review of the management of symptomatic stent-induced pancreatic duct injury following stent placement for prevention of post-ERCP pancreatitis in eight patients with previously normal pancreatic ducts. Subsequent treatment included pancreatic sphincterotomy, balloon dilation of stricture, and placement of multiple 3 - 5-Fr soft polymer pancreatic stents. All patients showed improvement or resolution of pancreatic strictures. Five patients had resolution or substantial improvement of pain, one patient showed a fair response with repeated ERCPs, and two patients failed to respond and underwent total pancreatectomy with islet autotransplantation. Pancreatic duct stent-induced ductal injury with significant clinical consequences can occur with conventional polyethylene stents. Endoscopic therapy is moderately effective but some patients develop irreversible damage. Caution should be used when placing standard polyethylene stents in normal ducts. Further research is required to identify safer materials and configurations of pancreatic stents. PMID- 19904702 TI - Thalidomide-related vessel thrombosis in microsurgical free tissue transfer. AB - This article describes a 49-year-old man with lower gum cancer who received tumor ablation by an otolaryngologist and immediate reconstruction with microsurgical free tissue transfer. The thrombosis occurred at the arterial anastomotic site at least twice during operation by an experienced microsurgeon and progressed to skin paddle necrosis following surgery. The thalidomide-induced recipient vessel thrombosis is highly suspected after excluding other hypercoagulation problems and technique errors. While thalidomide is increasingly used as an adjuvant therapy agent in head and neck cancer treatment, its potential adverse effect resulting in recipient vessel thrombosis is worth consideration in microsurgery. Prophylatic antithrombosis agents are thus recommended to prevent this potential side effect. PMID- 19904703 TI - Distally based posterior tibial artery cross-bridge flap for reconstruction of contralateral leg soft tissue defects. AB - A cross-leg or cross-bridge free flap is frequently used for the reconstruction of leg defects in cases with no suitable recipient vessels in the vicinity of the defect. Here we report eight cases of leg reconstruction with a distally based posterior tibial artery cross-bridge flap without microvascular anastomosis technology. A distally based posterior tibial artery flap was elevated and used to reconstruct contralateral leg defects. A skin incision was made near the recipient wound to create a skin bridge, and a subcutaneous tunnel was made between the skin incision and the recipient wound for the vascular pedicle to pass over. All flaps survived completely without complications, except for a case with minor erosion in the donor site. Compared with a cross-leg or cross-bridge free flap, a distally based posterior tibial artery cross-bridge flap is reliable and versatile, leading to shorter operation duration and a lower risk of thrombosis in the vessel because microvascular anastomosis is not required. PMID- 19904704 TI - [Why sexuality stays enigmatic]. PMID- 19904705 TI - [Drop-out from after-care following compulsory hospitalisation--how serious is the problem?]. AB - BACKGROUND: Little is known about the amount and quality of after-care provided for patients hospitalised as a result of a court authorisation. AIM: To obtain insight into the rate of drop-out from after-care and the quality of after-care. METHOD: In our study we included all patients in Rotterdam Rijnmond who, in the last 3 months of 2004, had been compulsorily hospitalised for at least one day by reason of a court authorisation. A retrospective study of patients' records let us ascertain whether drop-out from after-care occurred and let us check on the quality of the after-care provided. RESULTS: 214 patients were included. Of these, 33 (15.4%) dropped out of after-care. Prior to discharge, the drop-out group received an outpatient appointment at a local clinic less often and waited longer for their first appointment at that clinic than did the 'non-drop-out' group. The medical records of the drop-out group were less accurate and there was less cooperation between community care and clinical mental health care professionals. CONCLUSION: The quality of after-care can be improved if community care and clinical health care professionals cooperate more intensively in drawing up conditions for discharge and in arranging the transfer of patients from clinical care to community care. Further investigations are needed to find out whether these steps will have a beneficial effect on the drop-out percentage. PMID- 19904706 TI - [Addiction and recidivism in forensic psychiatry]. AB - BACKGROUND: The prevalence of addiction in forensic psychiatric patients (adolescents and adults) is unknown. Published figures are nearly always based on estimates or on old or select samples. AIM: To determine the importance of addiction as a risk factor for recidivism in forensic psychiatric patients in the Netherlands and to determine the prevalence of addiction in this group and the prevalence of drug intoxication in group-members at the time they committed the offence. METHOD: We analysed one database relating to forensic psychiatric adolescents and two sets of data relating to tbs-patients (tbs='detained at her Majesty's pleasure'). RESULTS: The prevalence of addiction among forensic psychiatric patients could only be given approximately, but it was certainly high (more than 60%). Substance abuse or addiction often played a major role (in more than 30% of cases) at the time the offence was committed for which the detention order was issued. CONCLUSION: Because substance abuse often played a role in the delinquent behaviour that led to the detention order, it may also be a risk factor for recidivism. More research is needed so that we can increase our knowledge about the most effective ways of treating substance abuse. PMID- 19904707 TI - [Physical training to treat depression]. AB - BACKGROUND: According to the Dutch multidisciplinary guideline of 2005 on depression, physical training (including running therapy) is an effective method for treating depression. However, very few reports have appeared about how this therapy should be applied and so far the intervention does not seem to be in general use. AIM: To review the literature from 2001 onwards relating to the extent and method of applying intensive physical training to the treatment of depression. METHOD: We searched various databases for articles published since 2001 on the effect of physical training on symptoms of depression and on possible underlying mechanisms. Articles were selected for their relevance according to title and abstract. RESULTS: The selected articles (n=51) seemed to show that physical training does reduce the symptoms of depression and they provided strong evidence that physical training markedly reduces the chance of subsequent somatic illnesses. We found two recent meta-analyses, one recent well-organised study showing only a slight effect, two later studies showing a dose-response relationship and three studies forming part of an addition strategy. There are interesting neurobiological explanatory models. If health professionals are thinking of introducing physical training for a patient they must realise that the patient will require close supervision by an expert. There are risks involved in making a patient participate in intensive physical exercise. CONCLUSION: Physical training is a valuable part of a stepped-care programme for treating patients with mild or moderate depression. PMID- 19904708 TI - [Serotonin-noradrenalin reuptake inhibitors in the treatment of non-malignant pain syndromes; a systematic review]. AB - BACKGROUND: Chronic pain and depressive symptoms are often comorbid. Antidepressants seem to influence not only the symptoms of depression but also the perception of pain. AIM: To give a systematic overview of the efficacy and safety of serotonin-noradrenalin reuptake inhibitors (SNRI) in the treatment of chronic non-malignant pain syndromes. METHODS: We reviewed the literature by means of PubMed and PsycInfo using combinations of the words 'pain' 'venlafaxine' and 'duloxetine'. We selected clinical studies that investigated the influence of SNRIs on pain perception. RESULTS: Fourteen articles met our selection criteria. Medical conditions involved were fibromyalgia, diabetic neuropathy and post mastectomy pain. Twelve studies demonstrated the efficacy and safety of venlafaxine and duloxetine in the treatment of non-malignant pain. CONCLUSION: The results revealed that SNRIs are effective in reducing pain particularly in the treatment of diabetic neuropathy. Results with regard to the other medical conditions are less clear. More research is needed to find out in which medical conditions SNRIs have a significant pain reducing effect and why this effect does not hold in the case of other medical conditions. PMID- 19904709 TI - [Becoming a psychiatrist: an intensive induction course for trainee house officers]. AB - Since October 2006 the specialist training for house officers in psychiatry in the province of North Holland has started with a one-week intensive induction course. The course has been designed to facilitate the transition to specialist training and to provide trainees with the tools they need for dealing with psychiatric emergencies. Fifteen house officers, divided into three focus groups, were asked for their views on the course. They gave the course a high rating and stated that it had had a positive effect on group-bonding and functioning in their subsequent training. PMID- 19904710 TI - [Frontal syndrome resulting from an intracerebral amyloidoma]. AB - A 51-year-old woman suffering an epileptic seizure came to the emergency unit. A ct scan showed a mass lesion in the right frontal lobe. The psychiatric examination indicated a frontal syndrome with severe cognitive impairment. A stereotactical biopsy was carried out. Histopathology produced an unusual diagnosis, namely an intracerebral amyloidoma. By reviewing the literature on intracerebral amyloidoma we were able to compare the psychiatric symptoms of published cases with those of this case. PMID- 19904711 TI - [Music as an uninvited guest: the auditive variant of the Charles Bonnet syndrome]. AB - Visually handicapped patients can be tormented by complex visual hallucinations (Charles Bonnet syndrome). Likewise, deaf patients and patients with impaired hearing can be plagued by auditory hallucinations, mostly involving music. Our article focuses on three female patients who suffered from musical hallucinations. In one of these patients the hallucinations ceased when her hearing was restored. In the second patient the hallucinations ceased when carbamazepine was prescribed. Quetiapine reduced the musical hallucinations in the third patient. The differential diagnoses and therapeutic options are discussed. PMID- 19904712 TI - [Cannabis-induced mania? A case study and literature review]. AB - A 22-year-old man with a long history of cannabis-use presented with a full blown mania. According to recent population-based studies, someone who starts using cannabis early in life runs the risk of developing not only psychosis but also manic symptoms and bipolar disorder. Further literature research showed that the use of cannabis influences both the onset and the course of bipolar disorder. When a patient presents with mania it might be advisable for the psychiatrist to take cannabis-use into account when making a diagnosis and prescribing treatment. PMID- 19904713 TI - Nicotinoid and pyrethroid insecticide resistance in houseflies (Diptera: Muscidae) collected from Florida dairies. AB - BACKGROUND: The housefly, Musca domestica L., continues to be a major pest of confined livestock operations. Houseflies have developed resistance to most chemical classes, and new chemistries for use in animal agriculture are increasingly slow to emerge. Five adult housefly strains from four Florida dairy farms were evaluated for resistance to four insecticides (beta-cyfluthrin, permethrin, imidacloprid and nithiazine). RESULTS: Significant levels of tolerance were found in most field strains to all insecticides, and in some cases substantial resistance was apparent (as deduced from comparison with prior published results). At the LC(90) level, greater than 20-fold resistance was found in two of the fly strains for permethrin and one fly strain for imidacloprid. Beta-cyfluthrin LC(90) resistance ratios exceeded tenfold resistance in three fly strains. The relatively underutilized insecticide nithiazine had the lowest resistance ratios; however, fourfold LC(90) resistance was observed in one southern Florida fly strain. Farm insecticide use and its impact on resistance selection in Florida housefly populations are discussed. CONCLUSION: Housefly resistance to pyrethroids is widespread in Florida. Imidacloprid resistance is emerging, and tolerance was observed to both imidacloprid and nithiazine. If these insecticides are to retain efficacy, producer use must be restrained. PMID- 19904714 TI - Potential of piperonyl butoxide-synergised pyrethrins against psocids (Psocoptera: Liposcelididae) for stored-grain protection. AB - BACKGROUND: Piperonyl butoxide (PB)-synergised natural pyrethrins (pyrethrin:PB ratio 1:4) were evaluated both as a grain protectant and a disinfestant against four Liposcelidid psocids: Liposcelis bostrychophila Badonnel, L. entomophila (Enderlein), L. decolor (Pearman) and L. paeta Pearman. These are key storage pests in Australia that are difficult to control with the registered grain protectants and are increasingly being reported as pests of stored products in other countries. Firstly, mortality and reproduction of adults were determined in wheat freshly treated at 0.0, 0.75, 1.5, 3 and 6 mg kg(-1) of pyrethrins + PB (1:4) at 30 +/- 1 degrees C and 70 +/- 2% RH. Next, wheat treated at 0.0, 1.5, 3 and 6 mg kg(-1) of pyrethrins + PB (1:4) was stored at 30 +/- 1 degrees C and 70 +/- 2% RH and mortality and reproduction of psocids were assessed after 0, 1.5, 3 and 4.5 months of storage. Finally, the potential of synergised pyrethrins as a disinfestant was assessed by establishing time to endpoint mortality for adult psocids exposed to wheat treated at 3 and 6 mg kg(-1) of synergised pyrethrins after 0, 3, 6, 9 and 12 h of exposure. RESULTS: Synergised pyrethrins at 6 mg kg( 1) provided 3 months of protection against all four Liposcelis spp., and at this rate complete adult mortality of these psocids can be achieved within 6 h of exposure. CONCLUSION: Piperonyl butoxide-synergised pyrethrins have excellent potential both as a grain protectant and as a disinfestant against Liposcelidid psocids. PMID- 19904715 TI - On-line solid-phase extraction coupled with liquid chromatography/electrospray ionization mass spectrometry for the determination of trace tributyltin and triphenyltin in water samples. AB - On-line solid-phase extraction (SPE) for pre-concentration and sample cleanup is one strategy to reduce matrix effects and to simultaneously improve detection sensitivity in liquid chromatography/mass spectrometry (LC/MS). This paper describes an on-line SPE-LC/MS method for the determination of tributyltin (TBT) and triphenyltin (TPhT) at trace levels in water samples. The direct coupling of an on-line C18 pre-column to LC/MS was used to pre-concentrate TBT and TPhT at trace levels from waters and to remove interfering matrix effects. Pre concentration was followed by separation of TBT and TPhT on a C18 column using a mobile phase containing 0.1% (v/v) HCOOH/5 mM HCOONH4 and methanol. While both electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) can be interfaced with MS for the detection of TBT and TPhT, ESI-MS was preferred for this application. The calibration curve for the targets was linear in the concentration range 0.1-30 microg L(-1). The detection limit (signal-to-noise (S/N) ratio = 3) was 0.02 microg L(-1) when 3.0 mL of sample was enriched on the C18 pre-column. The recoveries of TBT and TPhT in spiked waters were from 81.0 to 101.9%. The reproducibilities for the analysis of the standard mixture (10 microg L(-1)) for TBT and TPhT were 13.1 and 5.0%, respectively. The developed method was an easy and fast way to analyze TBT and TPhT in water samples. PMID- 19904716 TI - Development of a method to measure methadone enantiomers and its metabolites without enantiomer standard compounds for the plasma of methadone maintenance patients. AB - A liquid chromatography-photodiode array (LC-PDA) method using a chiral analytical column was developed to determine the plasma levels of enantiomers of methadone and its chiral metabolite, 2-ethylidene-1,5-dimethyl-3,3 diphenylpyrrolidine (EDDP), without the standard compounds of R-form or S-form enantiomers. This method was established by the characteristics of recombinant cytochrome P-450 (CYP) isozymes, where CYP2C19 prefers to metabolize R-methadone and CYP2B6 prefers to metabolize S-methadone. We incubated the racemic methadone standard with either enzyme for 24 h. We identified the retention times of R- and S-methadone to be around 10.72 and 14.46 min, respectively. Furthermore, we determined the retention times of R- and S-EDDP to be approximately 6.76 and 7.72 min, respectively. No interferences were shown through the retention times of morphine, buprenorphine and diazepam. With the high recovery rate of a solid phase extraction procedure, this method was applied in analyzing plasma concentrations of seven methadone maintenance patients where R- and S-methadone and R- and S-EDDP were 233.4 +/- 154.9 and 185.9 +/- 136.3 ng/mL and 84.4 +/- 99.4 and 37.6 +/- 22.9 ng/mL, respectively. These data suggest that the present method can be applied for routine assay for plasma methadone and EDDP concentrations for patients under treatment. PMID- 19904717 TI - Factors controlling the activity of the SERCA2a pump in the normal and failing heart. AB - Heart failure is the leading cause of death in western countries and is often associated with impaired Ca(2+) handling in the cardiomyocyte. In fact, cardiomyocyte relaxation and contraction are tightly controlled by the activity of the cardiac sarco(endo)plasmic reticulum (ER/SR) Ca(2+) pump SERCA2a, pumping Ca(2+) from the cytosol into the lumen of the ER/SR. This review addresses three important facets that control the SERCA2 activity in the heart. First, we focus on the alternative splicing of the SERCA2 messenger, which is strictly regulated in the developing heart. This splicing controls the formation of three SERCA2 splice variants with different enzymatic properties. Second, we will discuss the role and regulation of SERCA2a activity in the normal and failing heart. The two well-studied Ca(2+) affinity modulators phospholamban and sarcolipin control the activity of SERCA2a within a narrow window. An aberrantly high or low Ca(2+) affinity is often observed in and may even trigger cardiac failure. Correcting SERCA2a activity might therefore constitute a therapeutic approach to improve the contractility of the failing heart. Finally, we address the controversies and unanswered questions of other putative regulators of the cardiac Ca(2+) pump, such as sarcalumenin, HRC, S100A1, Bcl-2, HAX-1, calreticulin, calnexin, ERp57, IRS-1, and -2. PMID- 19904718 TI - Renal glutathione transport: Identification of carriers, physiological functions, and controversies. AB - Glutathione (GSH) is an endogenous tripeptide composed of the amino acids L glutamate, L-cysteine, and glycine. It is found in virtually all aerobic cells and plays critical roles in maintenance of cellular redox homeostasis and drug metabolism. An important component of its regulation is transport across biological membranes. Because GSH is a charged, hydrophilic molecule, transport occurs via catalysis by specific carrier proteins rather than by simple diffusion. Although it has been clearly understood that efflux of GSH across membranes such as the canalicular and sinusoidal plasma membranes in hepatocytes and the brush-border plasma membrane in renal proximal tubules is a key step in GSH turnover and interorgan metabolism, the existence and physiological functions of uptake of GSH across various epithelial plasma membranes has been subject to some debate. Besides transport across plasma membranes, GSH transport across intracellular membranes, most notably the mitochondrial inner membrane, has received some attention in recent years because of the importance of mitochondrial redox status and the mitochondrial GSH pool in cellular physiology and pathology. This commentary will focus on renal transport processes for GSH and will discuss some of the controversies that have existed and still seem to exist in the literature, specifically regarding uptake of intact GSH by basolateral membranes of renal proximal tubular cells and uptake of intact GSH by the mitochondrial inner membrane. PMID- 19904719 TI - CD40/CD40L signaling and its implication in health and disease. AB - CD40, a transmembrane receptor of the tumor necrosis factor gene superfamily is expressed on a variety of cells, such as monocytes, B-cells, antigen presenting cells, endothelial, smooth muscle cells, and fibroblasts. The interaction between CD40 and CD40 ligand (CD40L) enhances the expression of cytokines, chemokines, matrix metalloproteinases, growth factors, and adhesion molecules, mainly through the stimulation of nuclear factor kappa B. The aim of this review is to summarize the molecular and cellular characteristics of CD40 and CD40L, the mechanisms that regulate their expression, the cellular responses they stimulate and finally their implication in the pathophysiology of inflammatory and autoimmune diseases. PMID- 19904720 TI - High-performance liquid chromatography with solid-phase extraction for the quantitative determination of nilotinib in human plasma. AB - A simple, rapid and sensitive high-performance liquid chromatography (HPLC)-based method with ultraviolet detection was developed for the quantitation of nilotinib, a tyrosine kinase inhibitor, in human plasma. Nilotinib and the internal standard dasatinib were separated using a mobile phase of 0.5% KH(2)PO(4) (pH2.5)-acetonitrile-methanol (55:25:20, v/v/v) on a Capcell Pak MG II column (250 x 4.6 mm) at a flow rate of 0.5 mL/min and optical measurement at 250 nm. Analysis required only 100 microL of plasma and involved a rapid and simple solid-phase extraction with an Oasis HLB cartridge, which gave recoveries from 72 to 78% for nilotinib and from 74 to 76% for dasatinib. The lower limit of quantification for nilotinib was 10 ng/mL. The linear range of this assay was between 10 and 5000 ng/mL (r(2) > 0.9992 for the regression line). Intra- and inter-day coefficients of variation were less than 10.0% and accuracies were within 10.4% over the linear range. Our results indicate that this method is applicable to the monitoring of plasma levels of nilotinib in a clinical setting. PMID- 19904721 TI - Xylan-based nanoparticles: prodrugs for ibuprofen release. AB - Esterification of xylan with ibuprofen via activiation of the carboxylic acid with N,N'-carbonyldiimidazole (CDI) yields products of high drug loadings. Subsequent sulfation of xylan ibuprofen esters using the gentle agent SO(3)/DMF was successfully carried out in order to modify hydrophobicity of the xylan esters. The structure of the novel xylan esters was evaluated by means of NMR spectroscopy. The resulting xylan derivatives self assemble into spherical nanoparticles with mean diameters ranging from 162 to 472 nm. Preliminary stability measurements indicate that hydrolytic stability decreases with increase in degree of substitution of sulfate groups. Thus, a new concept toward improved drug delivery from polysaccharide-based nanoparticles can be established here. PMID- 19904722 TI - Amphiphilic block copolymers bearing ortho ester side-chains: pH-dependent hydrolysis and self-assembly in water. AB - A new type of pH-responsive block copolymer nanoparticle has been synthesized and characterized. The amphiphilic diblock copolymer, PEG-b-PMYM, contains acid labile ortho ester side-chains in the hydrophobic block and can self-assemble into micelle-like nanoparticles in water at neutral pH. Hydrolysis of the ortho ester side-chains follows a distinct exocyclic mechanism and shows pH-dependent kinetics, which triggers changes in nanoparticle size and morphology. The nanoparticles have been found to be non-toxic to cells in vitro. The ability to tune the size and morphology of biocompatible block copolymer nanoparticles by controlling the pH-sensitive side-chain hydrolysis represents a unique approach that may be exploited to improve the efficacy of nanometer-scale drug delivery. PMID- 19904723 TI - A novel temperature-responsive polymer as a gene vector. AB - A temperature-responsive polymer poly{2-(dimethylamino)ethyl methacrylate-co-[cis butenedioic anhydride-poly[(N-isopropylacrylamide)-co-(butyl methacrylate)]]} (PDMNIB) was synthesized by free radical polymerization. The polymer had a significant temperature-responsive behavior with a lower critical solution temperature (LCST) at 20 degrees C. Gel retardation assay showed that PDMNIB could efficiently interact with DNA. Dynamic light scattering (DLS) and zeta potential measurement indicated that the average sizes and the surface electric charges of the PDMNIB/DNA complexes could be changed by temperature. Due to the thermosensitive interaction between PDMNIB and DNA, the gene transfection efficiency of PDMNIB could be improved by temperature. PMID- 19904724 TI - Reduction-sensitive reversibly crosslinked biodegradable micelles for triggered release of doxorubicin. AB - Reduction-responsive reversibly crosslinked biodegradable micelles were developed and applied for triggered release of doxorubicin (DOX). An amphiphilic block copolymer of poly(ethylene glycol) (PEG) and poly(epsilon-caprolactone) (PCL) that contains two lipoyl functional groups at their interface (PEG-L(2)-PCL) has been synthesized. (1)H NMR spectroscopy and gel permeation chromatography (GPC) measurements show that the PEG-L(2)-PCL block copolymer had a controlled composition (PEG 5 kDa and PCL 5.4 kDa) and a polydispersity index (PDI) of 1.36. PEG-L(2)-PCL formed micelles with sizes that ranged from 20 to 150 nm in aqueous solutions, wherein a critical micelle concentration (CMC) of 16 mg.L(-1) was determined. The micelles were readily crosslinked by adding 7.6 mol % of dithiothreitol (DTT) relative to the lipoyl groups. Notably, micelles after crosslinking demonstrated a markedly enhanced stability against dilution, physiological salt concentration, and organic solvent. In the presence of 10 x 10(-3) M DTT, however, micelles were subject to rapid de-crosslinking. In vitro release studies showed minimal release of DOX from crosslinked micelles at a concentration of 10 mg L(-1) (C < CMC, analogous to intravenous injection), wherein less than 15% of the DOX was released in 10 h. In contrast, rapid release of DOX was observed for DOX-loaded non-crosslinked micelles under otherwise the same conditions ( approximately 80% release in 0.5 h). In the presence of 10 x 10(-3) M DTT mimicking an intracellular reductive environment, sustained release of DOX from crosslinked micelles was achieved, in which 75% of the DOX was released in 9 h. These novel reduction-sensitive reversibly crosslinked biodegradable micelles are highly promising for targeted intracellular delivery of anticancer drugs. PMID- 19904726 TI - Functional L-lysine dendritic macromolecules as liver-imaging probes. AB - Liver-imaging probes are prepared through the conjugation of Gd chelates and galactosyl moieties to peptide dendrimers. The dendritic probes possessing highly controlled structures and a single molecular weight have a two-fold increase in T(1) relaxivity to 9.1 x 10(3) (Gd M)(-1) s(-1) compared to Gd-DTPA. No obvious cytotoxicity of this multifunctional dendritic agent is discovered in vitro. The dendrimer bearing galactosyl moieties leads to a much-higher hepatocyte-cell uptake in vitro and provides good signal-intensity enhancement (35%) of mouse liver in vivo especially at 60 min after intravenous injection. In comparison, non-targeting Gd dendrimers provide only an 11% enhancement of imaging contrast at the same time point. Overall, the dendrimers bearing galactosyl moieties may be used as liver-imaging probes. PMID- 19904725 TI - Water soluble polymer protected lipofectamine 2000/DNA complexes for solid-phase transfection. AB - A fast degrading cholic acid-functionalized star poly(DL-lactide) has been used to fabricate polymer films to support Lipofectamine 2000/DNA complexes for mediating solid-phase transfection. To improve the gene expression activity, a water-soluble polymer, poly-alpha,beta-[N-(2-hydroxyethyl)-L-aspartamide (PHEA), was added to protect the complexes. The in vitro gene transfection in 293T cells, HeLa cells, and 3T3 cells showed that the gene expressions could be effectively mediated by the deposited Lipofectamine 2000/DNA complexes encapsulated in polymer films. The degradation of the polymer films that occurred during gene transfection did not show any unfavorable effects on the gene expression. PMID- 19904727 TI - Performance evaluation of a 32-element head array with respect to the ultimate intrinsic SNR. AB - The quality of an RF detector coil design is commonly judged on how it compares with other coil configurations. The aim of this article is to develop a tool for evaluating the absolute performance of RF coil arrays. An algorithm to calculate the ultimate intrinsic signal-to-noise ratio (SNR) was implemented for a spherical geometry. The same imaging tasks modeled in the calculations were reproduced experimentally using a 32-element head array. Coil performance maps were then generated based on the ratio of experimentally measured SNR to the ultimate intrinsic SNR, for different acceleration factors associated with different degrees of parallel imaging. The relative performance in all cases was highest near the center of the samples (where the absolute SNR was lowest). The highest performance was found in the unaccelerated case and a maximum of 85% was observed with a phantom whose electrical properties are consistent with values in the human brain. The performance remained almost constant for 2-fold acceleration, but deteriorated at higher acceleration factors, suggesting that larger arrays are needed for effective highly-accelerated parallel imaging. The method proposed here can serve as a tool for the evaluation of coil designs, as well as a tool to guide the development of original designs which may begin to approach the optimal performance. PMID- 19904728 TI - Tropane alkaloid profiling of hydroponic Datura innoxia Mill. Plants inoculated with Agrobacterium rhizogenes. AB - INTRODUCTION: Hydroponics has been shown as a possible way to produce high quality plant biomass with improved phytochemical levels. Nevertheless, effects of plant biotic and abiotic environment can lead to drastic changes and plant growth conditions must be optimised. OBJECTIVE: To evaluate how much microbes and Agrobacterium rhizogenes TR7 wild strain may affect the tropane alkaloid profile in Datura innoxia Mill. plants cultivated in hydroponic conditions. METHODOLOGY: Datura innoxia Mill. plants were cultivated in hydroponic with sterile or non sterile conditions. For half of the non-sterile plants, Agrobacterium rhizogenes TR7 strain was added to the nutrient solution for hydroponics. The tropane alkaloid content of leaves and roots was analysed by UFLC/ESI-HRMS and MS/MS. The metabolite profiles were compared using partial least square-discriminant analysis. RESULTS: In sterile conditions, aerial parts contained more scopolamine than the roots. However, the diversity of tropane alkaloids was greater in roots. Furthermore, 21 known compounds and four non-elucidated tropane alkaloids were found. The tropane alkaloid profile was shown to be statistically different between sterile and non-sterile hydroponic conditions. The levels of 3-acetoxy-6 hydroxytropane and 3-hydroxylittorine were higher in plants inoculated with A. rhizogenes. Five other tropane compounds were found in higher amounts in non axenic control plants. Hyoscyamine and scopolamine total contents were much higher in the whole plant co-cultivated with A. rhizogenes TR7 than in controls. Furthermore, the leaves and roots of axenic plants contained more alkaloids than non-sterile ones. CONCLUSION: In hydroponic conditions, microbes induced variations of the phytochemical levels. Addition of A. rhizogenes TR7 into the nutrient solutions improved the total hyoscyamine and scopolamine production. PMID- 19904729 TI - Enantioresolution of dl-penicillamine. AB - Penicillamine (PenA) is a nonproteinogenic amino acid containing a thiol group. The three functional groups in penicillamine undergo characteristic chemical reactions and differ in their ability to participate in various chemical and biochemical reactions. d-penicillamine is more active pharmacologically, while the l-isomer occurs 'naturally'. This review deals with the enantioresolution of PenA both by direct and indirect methods using liquid chromatography. HPLC separation of its diastereomers prepared with different chiral derivatizing reagents (on reversed-phase columns) and separation of the derivatives prepared with achiral reagents (on chiral columns or via ligand exchange mode) has been discussed. Separation of enantiomers tagged with achiral reagent (to add a chromophore for ehanced detection) when there is no diastereomer formation has been considered separately. In addition, application of PenA and its derivatives as chiral selector for enentioresolution of certain other compounds has also been discussed. PMID- 19904730 TI - Two-dimensional J-resolved NMR spectroscopy: review of a key methodology in the metabolomics toolbox. AB - One-dimensional (1D) (1)H NMR spectroscopy remains a leading analytical technology in metabolomics. Advantages of this approach include relatively rapid spectral acquisition and NMR resonances that provide a direct measure of metabolite concentration based upon a single internal standard. Severe spectral congestion can, however, significantly hinder both metabolite identification and quantification. Two-dimensional (1)H J-resolved (JRES) NMR spectroscopy retains many of the benefits of 1D NMR, but additionally disperses the overlapping resonances into a second dimension, reducing congestion and increasing metabolite specificity. The usefulness of this approach to metabolomics was first realised six years ago, and since then it has been used in biological, medical and environmental studies of plants and animals. Here we provide a basic introduction to the 2D JRES NMR experiment and then discuss strategies for spectral acquisition and processing in the context of metabolomics applications, concluding with some key recommendations: acquisition using a double spin-echo sequence with excitation sculpting; processing using the SEM window function, tilting and symmetricising, optionally followed by a skyline projection. Strategies for implementing JRES spectroscopy into the metabolomics toolbox are then considered, including its roles in metabolic fingerprinting, metabolite identification and metabolite quantification. Public resources and data standards for JRES metabolomics are reviewed. We conclude by evaluating the advantages (e.g. increased spectral dispersion and confidence in metabolite identification; fully automated processing; reduced batch-to-batch variation) and disadvantages (e.g. longer acquisition times; higher technical variability; phase-twisted lineshapes resulting in quantification errors) of 2D JRES NMR vs the established 1D approach for metabolomics. PMID- 19904731 TI - Application of NMR in plant metabolomics: techniques, problems and prospects. AB - The present state-of-the-art of NMR in plant metabolomics is reviewed. Attention is paid to the different practical aspects of the application of NMR. The sample preparation, the measurement of the spectrum, quantitative aspects and data analysis are discussed. Each stage has its specific problems, which are pointed out and recommendations are made. PMID- 19904732 TI - The photographer and the greenhouse: how to analyse plant metabolomics data. AB - INTRODUCTION: Plant metabolomics experiments yield large amounts of data, too much to be interpretable by eye. Multivariate data analyses are therefore essential to extract and visualise the information of interest. OBJECTIVE: Because multivariate statistical methods may be remote from the expertise of many scientists working in the metabolomics field, this overview provides a step-by step description of a multivariate data analysis, starting from the experiment and ending with the figures appearing in scientific journals. METHODOLOGY: We developed a thought experiment that explores the relationship between the differences in nutrient levels and three plant developmental descriptors through photography of the greenhouse they grow in. Through this, multivariate data analysis, data preprocessing and model validation are illustrated. Finally some of the presented methods are illustrated by the analysis of a plant metabolomics dataset. CONCLUSION: This paper will familiarize non-specialised researchers with the main concepts in multivariate data analysis and allow them to develop and evaluate metabolomic data analyses more critically. PMID- 19904733 TI - Sample preparation for plant metabolomics. AB - Sample preparation in plant metabolomics is a fundamental and critical step with important consequences for the accuracy of results. Depending on the analytical tools and the metabolites of interest, sample preparation has to be decided. However, the various methods reported in the literature have many steps in common and consequently the practical considerations concerning the pros and cons are similar. In this review, each step of the sample preparation - harvesting, drying, extraction and purification - will be discussed in detail. PMID- 19904734 TI - Differences between bisphosphonates in binding affinities for hydroxyapatite. AB - Bisphosphonates (BPs) inhibit bone resorption and are widely used for the treatment of bone diseases, including osteoporosis. BPs are also being studied for their effects on hydroxyapatite (HAP)-containing biomaterials. There is a growing appreciation that there are hitherto unexpected differences among BPs in their mineral binding affinities that affect their pharmacological and biological properties. To study these differences, we have developed a method based on fast performance liquid chromatography using columns of HAP to which BPs and other phosphate-containing compounds can adsorb and be eluted by using phosphate buffer gradients at pH 6.8. The individual compounds emerge as discrete and reproducible peaks for a range of compounds with different affinities. For example, the peak retention times (min; mean +/- SEM) were 22.0 +/- 0.3 for zoledronate, 16.16 +/- 0.44 for risedronate, and 9.0 +/- 0.28 for its phosphonocarboxylate analog, NE10790. These results suggest that there are substantial differences among BPs in their binding to HAP. These differences may be exploited in the development of biomaterials and may also partly explain the extent of their relative skeletal retention and persistence of biological effects observed in both animal and clinical studies. PMID- 19904735 TI - UV curable polyester polyol acrylate/bentonite nanocomposites: synthesis, characterization, and drug release. AB - Polyesterpolyolacrylate/bentonite nanocomposites, capable of in situ photo polymerization, were synthesized and characterized. The organically modified bentonite clay was prepared by an ion exchange process, in which sodium ions were replaced by alkyl ammonium ions. Organo modification of bentonite was confirmed from X-ray diffraction and fourier transform-infrared data. Microstructures were characterized by XRD data and transmission electron microscopy (TEM). Both XRD data and TEM images of polyester polyol acrylate/organo modified bentonite nanocomposites indicated that most of silicate layers were intercalated into the acrylate matrix. The resulting nanocomposites were characterized by gel content, water equilibrium swell, tensile strength, and in vitro degradation. The results showed that water equilibrium swell and in vitro degradation of these nanocomposites decreased with increase in the clay content. The tensile strength of these nanocomposites also increased with increase in the clay content. Release of two model drugs namely sulfamethoxazole and diclofenac sodium (DS) from these nanocomposites was studied in phosphate buffer saline pH = 7.4 at 37 degrees C. The drug release studies showed that sulfamethoxazole released slower than DS from polyester polyol acrylate nanocomposites. Therefore, these materials may be useful for mucoadhesive drug carriers and other biomedical applications. PMID- 19904736 TI - Gas chromatography coupled to electron capture negative ion mass spectrometry with nitrogen as the reagent gas--an alternative method for the determination of polybrominated compounds. AB - Gas chromatography in combination with electron capture negative ion mass spectrometry (GC/ECNI-MS) is a sensitive method for the determination of polybrominated compounds in environmental and food samples via detection of the bromide ion isotopes m/z 79 and 81. The standard reagent gas for inducing chemical ionization in GC/ECNI-MS is methane. However, the use of methane has some drawbacks as it promotes carbonization of the filament and ion source. In this study, we explored the suitability of nitrogen as reagent gas for the determination of brominated flame retardants (polybrominated diphenyl ethers (PBDEs), polybrominated biphenyls (PBBs), allyl-2,4,6-tribromophenyl ether (ATE) and 2,3-dibromopropyl-2,4,6-tribromophenyl ether (DPTE)) and halogenated natural products (for instance, methoxylated tetrabrominated diphenylethers and polybrominated hexahydroxanthene derivatives). An ion source temperature of 250 degrees C and a nitrogen pressure of 7 Torr in the ion source gave the highest response for m/z 79 and 81 of virtually all investigated polybrominated compounds. Using these conditions, nitrogen-mediated GC/ECNI-MS usually gave higher sensitivity than the method with methane previously used in our lab. In addition, the ion source was not contaminated to the same degree and the lifetime of the filament was significantly increased. Moreover, the response factors of the different polybrominated compounds with the exception of 2,4,6-tribromophenol were more uniform than with methane. Nitrogen is available at very high purity at relatively low price. PMID- 19904737 TI - Validation of pentaacetylaldononitrile derivative for dual 2H gas chromatography/mass spectrometry and 13C gas chromatography/combustion/isotope ratio mass spectrometry analysis of glucose. AB - A reference method to accurately define kinetics in response to the ingestion of glucose in terms of total, exogenous and endogenous glucose is to use stable isotope-labelled compounds such as 2H and 13C glucose followed by gas chromatography/mass spectrometry (GC/MS) and gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) analysis. The use of the usual pentaacetyl (5Ac) derivative generates difficulties in obtaining accurate and reproducible results due to the two chromatographic peaks for the syn and anti isomers, and to the isotopic effect occurring during acetylation. Therefore, the pentaacetylaldononitrile derivative (Aldo) was validated for both isotopes, and compared with the 5Ac derivative. A correction factor including carbon atom dilution (stoichiometric equation) and the kinetic isotopic effect (KIE) was determined. Analytical validation results for the 2H GC/MS and 13C GC/C/IRMS measurements produced acceptable results with both derivatives. When 2H enrichments of plasma samples were < or = 1 mol % excess (MPE), the repeatability (RSD(Aldo Intra assay and Intra day) <0.94%, RSD(5Ac Intra assay and Intra day) <3.29%), accuracy (Aldo <3.4%, 5Ac <29.0%), and stability of the derivatized samples were significantly better when the Aldo derivatives of the plasma samples were used (p < 0.05). When the glucose kinetics were assessed in nine human subjects, after glucose ingestion, the plasma glucose 2H enrichments were identical with both derivatives, whereas the 13C enrichments needed a correction factor to fit together. Due to KIE variation, this correction factor was not constant and had to be calculated for each batch of analyses, to obtain satisfactory results. Mean quantities of exogenous glucose exhibit marked difference (20.9 +/- 1.3g (5Ac) vs. 26.7 +/- 2.5g (Aldo)) when calculated with stoichiometric correction, but fit perfectly when calculated after application of the correction factor (22.1 +/- 1.3g (5Ac) vs. 22.9 +/- 1.9g (Aldo)). Finally, the pentaacetylaldononitrile derivative, used here in GC/C/IRMS for the first time, enables measurement of 2H and 13C enrichments in plasma glucose with a single sample preparation. PMID- 19904738 TI - Challenges of stem cell therapy for spinal cord injury: human embryonic stem cells, endogenous neural stem cells, or induced pluripotent stem cells? AB - Spinal cord injury (SCI) causes myelopathy, damage to white matter, and myelinated fiber tracts that carry sensation and motor signals to and from the brain. The gray matter damage causes segmental losses of interneurons and motoneurons and restricts therapeutic options. Recent advances in stem cell biology, neural injury, and repair, and the progress toward development of neuroprotective and regenerative interventions are the basis for increased optimism. This review summarizes the pathophysiological mechanisms following SCI and compares human embryonic, adult neural, and the induced pluripotent stem cell based therapeutic strategies for SCI. PMID- 19904739 TI - Improved biological performance of microarc-oxidized low-modulus Ti-24Nb-4Zr 7.9Sn alloy. AB - Ti-24Nb-4Zr-7.9Sn (TNZS) is a newly developed beta-titanium alloy with low modulus and it has been considered as good material for dental or orthopedic implant. The purpose of the current study is to evaluate the effect of micro-arc oxidation (MAO) treatment on the biological performance of TNZS surface. The phases, morphology and chemical composition of the MAO-treated surface were characterized by X-ray diffraction, energy dispersive spectroscope and scanning electron microscopy analysis respectively. Then we tested the biocompatibility by examining the cell morphology and viability of osteoblast cells growing on MAO TNZS surface. The bone binding strength of the specimens was evaluated by removal torque test after implantation in rabbit tibiae for 6 weeks. Compared with the none-treated titanium and TNZS specimens, MAO treated TNZS specimens showed a significant increase (p<0.05) in hydrophilicity, roughness, cell viability and removal torque forces. In summary, MAO treatment helps to form a porous surface with a biologically active bone-like apatite layer on TNZS specimens, which may improve the biological response of MAO-TNZS implants. PMID- 19904740 TI - Effect of growth factors in combination with injectable silicone resin particles on the biological activity of dermal fibroblasts: a preliminary in vitro study. AB - Injections of silicone fluid have been clinically evaluated to treat and prevent foot ulcers due to diminished plantar fat-pad in neuropathic diabetics. The objective of this study was to determine preliminary in vitro effects of an injectable form of silicone resin particles in combination with growth factors to determine the suitability of this potential therapy for prevention of diabetic foot ulcers. Basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) and platelet-derived growth factor (PDGF-BB) were added to monolayer culture along with silicone resin particles (12 microm average diameter). Growth factors were also combined as follows: bFGF+PDGF-BB, EGF+PDGF-BB, and bFGF+EGF. Growth factors alone and in combination increased fibroblast proliferation, but the presence of particles did not significantly affect cellular proliferation. The addition of particles significantly increased fibronectin production 117% in the control group and 151% in the PDGF only group. Collagen production was increased with exposure to EGF and growth factor combinations, but the presence of particles did not lead to any significant differences, except an 81% increase in the bFGF group. These preliminary results suggest that a combination of PDGF and bFGF may be effective in stimulating proliferation and matrix production around injectable silicone resin particles to generate a fibrous tissue pad to alleviate the abnormal distribution of high pressures that contribute to diabetic foot ulcer formation. PMID- 19904741 TI - Structural differences between apolipoprotein E3 and E4 as measured by (19)F NMR. AB - The apolipoprotein E family contains three major isoforms (ApoE4, E3, and E2) that are directly involved with lipoprotein metabolism and cholesterol transport. ApoE3 and apoE4 differ in only a single amino acid with an arginine in apoE4 changed to a cysteine at position 112 in apoE3. Yet only apoE4 is recognized as a risk factor for Alzheimer's disease. Here we used (19)F NMR to examine structural differences between apoE4 and apoE3 and the effect of the C-terminal domain on the N-terminal domain. After incorporation of 5-(19)F-tryptophan the 1D (19)F NMR spectra were compared for the N-terminal domain and for the full length proteins. The NMR spectra of the N-terminal region (residues 1-191) are reasonably well resolved while those of the full length wild-type proteins are broad and ill defined suggesting considerable conformational heterogeneity. At least four of the seven tryptophan residues in the wild type protein appear to be solvent exposed. NMR spectra of the wild-type proteins were compared to apoE containing four mutations in the C-terminal region that gives rise to a monomeric form either of apoE3 under native conditions (Zhang et al., Biochemistry 2007; 46: 10722-10732) or apoE4 in the presence of 1 M urea. For either wild-type or mutant proteins the differences in tryptophan resonances in the N-terminal region of the protein suggest structural differences between apoE3 and apoE4. We conclude that these differences occur both as a consequence of the Arg158Cys mutation and as a consequence of the interaction with the C-terminal domain. PMID- 19904742 TI - SYUNZ-16, a newly synthesized alkannin derivative, induces tumor cells apoptosis and suppresses tumor growth through inhibition of PKB/AKT kinase activity and blockade of AKT/FOXO signal pathway. AB - Alkannin is the major bioactive compound of Arnebia euchroma roots, which is used in many therapeutic remedies in Chinese traditional medicine. SYUNZ-16 is a new derivative of alkannin. In this study, anticancer effects of SYUNZ-16 on human lung adenocarcinoma cell line GLC-82 and human hepatocarcinoma cell line Hep3B were tested in vitro. The results showed SYUNZ-16 could obviously inhibit the proliferation of these cancer cell lines via induction of apoptosis, with the evidence of increasing AnnexinV-positive cells and cleaved caspase-3 and PARP fragments. More importantly, we found that SYUNZ-16 could inhibit AKT activity in cell-free system. Treatment of cancer cells with SYUNZ-16 decreased the phosphorylation of AKT. Additionally, SYUNZ-16 partially attenuated the phosphorylation levels of FKHR and FKHRL1 in a dose-dependent and time-dependent fashion, and led to an increase in the nuclear accumulation of exogenous FKHR, and upregulated the mRNA expression of Bim and TRADD in cancer cells. Further study showed that constitutively activated AKT1 transfection could reduce apoptosis induction mediated by SYUNZ-16. The in vivo experiments showed that SYUNZ-16 had inhibitory effects on S-180 sarcoma implanted to mice. And in GLC-82 xenograft models, SYUNZ-16 at 20 mg/kg/qod remarkably inhibited the tumor growth with the T/C value of 45.3%. Taken together, SYUNZ-16 might be a potent inhibitor of AKT signaling pathway in tumor cells. These data provide evidence for the development of SYUNZ-16 as a potential antitumor drug candidate for further research and development. PMID- 19904743 TI - High expression of EZH2 is associated with tumor aggressiveness and poor prognosis in patients with esophageal squamous cell carcinoma treated with definitive chemoradiotherapy. AB - The enhancer of zeste homolog 2 (EZH2), a known repressor of gene transcription, has been reported to be associated with biological malignancy in several cancers. The potential oncogenic role of EZH2 and its clinical/prognostic significance, however, in esophageal squamous cell carcinoma (ESCC) are unclear. In this study, the methods of immunohistochemistry and fluorescence in-situ hybridization were used to examine protein expression and amplification of EZH2 in 98 pretreatment biopsy specimens of ESCC who received definitive chemoradiotherapy (CRT). High expression of EZH2 and amplification of EZH2 was found in 54.1% and 12.0% of ESCCs, respectively. High EZH2 expression was significantly correlated with increased cell proliferation (p = 0.009), high histopathological grade (p = 0.002), regional (p = 0.025) and distant lymph node metastasis (p < 0.001) and lack of clinical complete response to CRT (p = 0.028). Univariate analysis revealed that high expression of EZH2 was associated with poor metastasis-free survival (MFS) (p = 0.003), poor progression-free survival (PFS) (p = 0.001) and poor disease-specific survival (DSS) (p < 0.001). In multivariate analysis, high expression of EZH2, together with lack of clinical complete response, were evaluated as significant independent prognostic factors of MFS, PFS and DSS for patients with ESCC. These findings suggest that high expression of EZH2 correlates with tumor aggressiveness and adverse patient outcome in ESCC treated with definitive CRT. Evaluation of EZH2 expressions might be useful for predicting tumor response to CRT and prognosis for patients with ESCC. PMID- 19904744 TI - N-glycan based models improve diagnostic efficacies in hepatitis B virus-related hepatocellular carcinoma. AB - The early diagnosis of hepatocellular carcinoma (HCC) is of great clinical desirable due to lack of specific and sensitive markers. Alterations in the sugar chains of glycoprotein synthesized by the liver contribute to the molecular basis of abnormalities in carcinogenesis. This study aims to construct and assess the diagnostic value of N-glycan based diagnostic model in HCC identification and follow-up. A total of 393 subjects including HBV-related HCC, liver fibrosis and healthy controls were recruited. Follow-up was carried out before and after surgical treatment in HCC. N-glycome of serum glycoprotein was profiled by DNA sequencer-assisted fluorophore-assisted carbohydrate electrophoresis (DSA-FACE). Multiparameters diagnostic models were constructed based on N-glycan markers. The result found that 2 N-glycan structure abundances (NG1A2F, Peak 4; NA3Fb, Peak 9) were useful as N-glycan markers. The diagnostic efficacy of the log ratio [log(p9/4)] was similar to that of AFP in differentiating HCC from fibrosis. The accuracy and sensitivity of the diagnostic model combining AFP and N-glycan markers (Cscore B) were increased 7-10% compared with that of AFP. Log(p9/4) was more efficient in monitoring the progression of HCC with regarding to vascular invasion at improved specificity (16%) and accuracy (8%) compared with that of AFP. The N-glycan markers were found to be changed significantly after surgical resection in HCC follow-up. We conclude that the branching alpha (1,3) fucosylated triantennary glycan and a biantennary glycan are promising as N glycan markers. The diagnostic models based on the N-glycan markers and AFP improve the efficacy in HCC diagnosis and progression monitoring. PMID- 19904745 TI - Tumor inhibition by sodium selenite is associated with activation of c-Jun NH2 terminal kinase 1 and suppression of beta-catenin signaling. AB - Epidemiological and clinical studies suggest that an increased intake of dietary selenium significantly reduces overall cancer risk, but the anticancer mechanism of selenium is not clear. In this study, we fed intestinal cancer mouse model. Muc2/p21 double mutant mice with a selenium-enriched (sodium selenite) diet for 12 or 24 weeks, and found that sodium selenite significantly inhibited intestinal tumor formation in these animals (p < 0.01), which was associated with phosphorylation of JNK1 and suppression of beta-catenin and COX2. In vitro studies showed that sodium selenite promoted cell apoptosis and inhibited cell proliferation in human colon cancer cell lines HCT116 and SW620. These effects were dose- and time course-dependent, and were also linked to an increase of JNK1 phosphorylation and suppression of beta-catenin signaling. Reduced JNK1 expression by small RNA interference abrogated sufficient activation of JNK1 by sodium selenite, leading to reduced inhibition of the beta-catenin signaling, resulting in reduced efficacy of inhibiting cell proliferation. Taken together, our data demonstrate that sodium selenite inhibits intestinal carcinogenesis in vivo and in vitro through activating JNK1 and suppressing beta-catenin signaling, a novel anticancer mechanism of selenium. PMID- 19904746 TI - Secretin inhibits cholangiocarcinoma growth via dysregulation of the cAMP dependent signaling mechanisms of secretin receptor. AB - Secretin plays a key role in the regulation of normal cholangiocyte physiology via secretin receptor (SCTR). SCTR expression is upregulated during extrahepatic cholestasis induced by bile duct ligation and closely associated with cholangiocyte proliferative responses. Although well studied in normal cholangiocytes, the role of secretin and the expression of SCTR in the regulation of cholangiocarcinoma proliferation are unknown. In vitro, secretin (10(-7) M) displayed differential effects on normal cholangiocyte [H-69 and human intrahepatic biliary epithelial cell line (HIBEpiC)] and cholangiocarcinoma (Mz ChA-1, HuH-28, TFK-1, SG231, CCLP1 and HuCC-T1) cell lines as such secretin is mitogenic for normal cholangiocytes and antiproliferative for cholangiocarcinoma. As expected in normal cholangiocytes (HIBEpiC), secretin increased intracellular cyclic adenosine monophosphate (cAMP) levels. However, the effect of secretin on intracellular cAMP levels was suppressed in Mz-ChA-1 cells. Secretin-stimulated intracellular cAMP levels in Mz-ChA-1 were restored by pretreatment with pertussis toxin, suggesting that the receptor coupled to Galpha(i) rather than Galpha(s). SCTR expression was found to be downregulated in 4 of the 6 cholangiocarcinoma cell lines evaluated and in human cholangiocarcinoma biopsy samples. In vivo, secretin significantly inhibited the tumor size and more than doubled tumor latency, which was associated with a decrease in proliferating cell nuclear antigen and an increase in cleaved-caspase 3 expression levels. Our results demonstrate that secretin and/or the modulation of SCTR expression might have potential as a therapeutic tool in the treatment of cholangiocarcinoma. PMID- 19904747 TI - Association between polymorphisms in interleukin-17A and interleukin-17F genes and risks of gastric cancer. AB - Chronic inflammation is the hallmark of the pathogenesis of Helicobacter pylori induced gastric cancer. Interleukin (IL)-17A and IL-17F are inflammatory cytokines expressed by a novel subset of CD4+ Th cells and play critical function in inflammation and probably in cancer. We conducted a case-control study including 1,010 gastric cancer patients and 800 healthy controls to assess the association between IL-17A G197A and IL-17F A7488G polymorphisms and risk of gastric cancer. Genotypes were determined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing. Logistic regression and Cox-proportional hazards analyses were used to evaluate the associations between polymorphisms and gastric cancer susceptibility, clinicopathological features and survival. After adjusted for age and gender, IL 17F 7488GA and GG genotypes were associated with an increased risk of gastric cancer compared with AA genotype [OR 1.51, 95% confidence interval (CI): 1.22 1.87 for GA; OR 1.61, 95% CI: 1.03-2.51 for GG]. Further stratification analyses indicated that the effect of IL-17F 7488GA genotype was noteworthy in gastric cancer patients of noncardia, intestinal type, poorly and moderately differentiated, age older than 40, large tumor size and lymph node metastasis. IL 17A 197AG genotype was associated with increased risk of poorly differentiated, TNM I/II, age of 40-65-year subtypes of gastric cancer, but not with total gastric cancer risk (p = 0.098). No significant relationship was observed between polymorphisms and survival of gastric cancer patients. These findings suggest that polymorphism of IL-17F 7488 involved in susceptibility to gastric cancer, which also influenced certain subtypes according to clinicopathological features, whereas IL-17A 197 may be less relevant. PMID- 19904748 TI - Tumor angiogenesis and novel antiangiogenic strategies. AB - Angiogenesis is essential for the development and growth of tumors. It is a highly regulated process that requires cross-talk between signaling pathways at all stages of blood vessel development and tumor growth, from the recruitment of endothelial cells to vessel maturation. This review summarizes tumor angiogenesis and describes the key signaling pathways governing blood vessel development. The role of angiogenesis in various tumor types is discussed, but the focus is on invasive breast cancer, a disease that will affect approximately 182,000 women in the USA in 2008. Research efforts over the past decade have identified numerous potential, as well as proven therapies with activity in breast cancer. These include chemotherapeutics as well as therapies that inhibit specific angiogenic pathways known as targeted agents. Some of the data from single- and multitargeted antiangiogenic agents are described in this review. "Published 2008 Wiley-Liss, Inc. This article is a US Government work, and, as such, is in the public domain in the United States of America." PMID- 19904749 TI - The effect of focal adhesion kinase gene silencing on 5-fluorouracil chemosensitivity involves an Akt/NF-kappaB signaling pathway in colorectal carcinomas. AB - Multicellular resistance (MCR) is produced because multicellular spheroids (MCSs) are formed with a broad cell-cell connection when cultured in three-dimensions, which limits the clinical treatment efficacy in solid tumors. Focal adhesion kinase (FAK) plays an important role in apoptosis, survival and cell adhesion between cells and their extracellular matrix. In this study, we investigated the expressions of FAK, Akt and NF-kappaB in human colorectal cancer (CRC), and the effects of FAK gene silencing on MCSs formation and 5-fluorouracil (5-FU) chemosensitivity in colon carcinoma MCSs culture cells. In CRC samples, FAK, Akt and NF-kappaB were overexpressed. The positive expression of FAK correlated notably with lymph node metastasis and cellular differentiation. Positive expressions of Akt and NF-kappaB were significantly related to cellular differentiation and lymph node metastasis, respectively. Furthermore, positive expression of FAK correlated with that of Akt and NF-kappaB. The expression of FAK was inhibited significantly by a small hairpin RNA targeting FAK. Knockdown of FAK reversed the formation and aggregation of MCSs, significantly decreased the 50% inhibitory concentration of 5-FU, and markedly increased MCS culture cells apoptosis. These effects were associated with reduced levels of Akt and NF kappaB. These results indicate that suppressing FAK expression potentiated 5-FU induced cytotoxicity and contributed to its chemosensitizing effect by suppressing Akt/NF-kappaB signaling in colon carcinoma MCS culture cells. These data also imply that FAK mediates MCR of CRC through the survival signaling pathway FAK/Akt/NF-kappaB. PMID- 19904750 TI - Latitudinal variation in cranial dimorphism in Macaca fascicularis. AB - This study examines latitudinal and insular variation in the expression of sexual dimorphism in cranial length in three geographical groupings of Macaca fascicularis. In addition, the relationship between cranial length dimorphism (CLD) and sex-specific size is examined. The results of the study identified a significant relationship between CLD and latitude for only one of the three geographic groupings. Sex-specific relationships between cranial length and CLD were detected. The pattern of these relationships varied by geographic grouping. This study is important because it demonstrates that despite very similar levels of CLD in a single primate species, there exists important geographic variability in the correlates of that dimorphism. I suggest that geographically varying ecological factors may influence sex-specific natural selection and the intensity of CLD in M. fascicularis. Gaining a better understanding of this geographical variability will require that future research examines morphological variation, including CLD, within its corresponding ecological and social contexts. Such research should be comparative, and incorporate multiple geographically separated populations with disparate environmental settings. PMID- 19904751 TI - Primate conservation: integrating communities through environmental education programs. AB - Environmental education has evolved over the years to respond to the varied complexities found in the different localities where it is practiced. In many parts of the world where biodiversity is rich, social conditions are poor, so educators have included sustainable development alternatives to better the environment and the livelihoods of local communities. Primate conservation education programs, which are often based in areas that face such challenges, have been a vanguard in creating means to integrate people with their natural environment and thus conquer supporters for the protection of natural habitats. In the search for effectiveness they have adopted evaluation methods to help assess what was offered. An example from Brazil is described in this commentary. PMID- 19904752 TI - Psychopathy and axis I psychiatric disorders among criminal offenders: relationships to impulsive and proactive aggression. AB - Both psychopathology and aggression are heterogeneous constructs. Determining which forms of psychopathology relate to risk for different classes of aggressive behavior has implications for risk recognition and management. This study examined the relationships of impulsive aggression (IA) and proactive aggression (PA) to psychopathy and symptoms of several Diagnostic and Statistical Manual Axis I disorders in a sample of criminal offenders. Results replicated prior findings from community samples of a broad relationship between psychopathology and IA. PA was related only to psychopathy. An interaction was found whereby IA was associated with impulsive-antisocial traits of psychopathy only for individuals with moderate to high levels of generalized anxiety. Results indicate that assessing and treating several Axis I disorders in offenders may decrease risk for IA. Moreover, current findings raise the possibility that generalized anxiety is a key, modifiable component of the relationship between IA and impulsive-antisocial traits. PMID- 19904753 TI - Autoantibody potential of cancer therapeutic monoclonal antibodies. AB - We and others have reported that multiple autoantibodies are unmasked in human polyclonal antibody preparations after exposure to physiological oxidizing agents (hemin) or electromotive force. We now have asked if oxidation unmasks autoantibody reactivities in monoclonal antibodies (mAb). To do this, we have studied 9 FDA approved mAb used therapeutically, including 4 chimeric, 4 humanized and 1 chemically modified chimeric Fab that were exposed to the physiological oxidizing agent hemin at 36 degrees C for 20 hr. These mAb were studied for autoantibody activity to phospholipids and DNA before and after oxidation with hemin and found to develop autoantibody activities after oxidation, while retaining their original specificity as measured by mAb anti glycophorin A binding of erythrocytes, CD 19 binding to B lymphocytes and anti HLA-A29 binding to A29-positive lymphocytes. The finding that certain mAb have the potential to unmask autoantibody activities as a consequence of exposure to physiological redox reactions in vitro gives pause to our present understanding of the immunological basis of tolerance and concern for potential autoimmune side effects in patients receiving mAb for diagnosis or treatment. PMID- 19904754 TI - Oxidative stress-dependent increase in ICAM-1 expression promotes adhesion of colorectal and pancreatic cancers to the senescent peritoneal mesothelium. AB - Intercellular adhesion molecule-1 (ICAM-1) has been implicated in adhesion of colorectal and pancreatic cancer cells (of the SW480 and PSN-1 line, respectively) to the peritoneal mesothelium. It has been demonstrated that ICAM-1 expression increases with senescence in some cell types, however, the significance of this phenomenon in the context of malignant dissemination remains elusive. In this report we show that the adherence of SW480 and PSN-1 cells to senescent human omentum-derived mesothelial cells (HOMCs) in vitro is greater than to early-passage cells and that the effect is mediated by ICAM-1. Senescent HOMCs display increased expression of ICAM-1 mRNA and cell surface protein. The development of this phenotype is related to increased oxidative stress in senescent cells. The augmented ICAM-1 expression in HOMCs can be reduced by culturing cells with antioxidants; in contrast, exposure of HOMCs to an oxidant, t-BHP, leads to cellular senescence and increased ICAM-1 expression. The effect is partly mediated by activation of p38 MAPK and AP-1 signaling pathways. Finally, culture of HOMCs in the presence of a strong antioxidant, PBN, significantly reduces the senescence-associated increase in SW480 and PSN-1 cancer cell binding. These results indicate that increased oxidative stress and increased expression of ICAM-1 in senescent HOMCs may facilitate peritoneal adhesion of selected colorectal and pancreatic cancers. PMID- 19904755 TI - GBV-C/hepatitis G virus infection and non-Hodgkin lymphoma: a case control study. AB - We investigated whether there was an association between GBV-C viremia and the development of non-Hodgkin lymphoma (NHL) in 553 NHL cases and 438 controls from British Columbia, Canada. Cases were aged 20-79, diagnosed between March 2000 and February 2004, and resident in Greater Vancouver or Victoria. Cases and controls were tested for GBV-C RNA by RT-PCR and positive samples were genotyped. Overall, GBV-C RNA was detected in 4.5% of NHL cases vs. 1.8% of controls [adjusted odds ratio (OR) = 2.72, 95% confidence interval (CI) = 1.22-6.69]. The association between GBV-C RNA detection and NHL remained even after individuals with a history of prior transfusion, injection drug use and hepatitis C virus sero positivity were excluded. GBV-C viremia showed the strongest association with diffuse large B cell lymphoma (adjusted OR = 5.18, 95% CI = 2.06-13.71). Genotyping was performed on 29/33 GBV-C RNA positive individuals; genotypes 2a (n = 22); 2b (n = 5) and 3 (n = 2) were identified, consistent with the distribution of genotypes found in North America. This is the largest case-control study to date associating GBV-C viremia and NHL risk. As GBV-C is known to be transmitted through blood products this may have important implications for blood safety. PMID- 19904756 TI - Establishment of the 1st World Health Organization international standards for human papillomavirus type 16 DNA and type 18 DNA. AB - A World Health Organization collaborative study was conducted to evaluate candidate international standards for human papillomavirus (HPV) Type 16 DNA (NIBSC code 06/202) and HPV Type 18 DNA (NIBSC code 06/206) for use in the amplification and detection steps of nucleic acid-based assays. The freeze-dried candidate international standards were prepared from bulk preparations of cloned plasmid containing full-length HPV-16 or HPV-18 genomic DNA. Nineteen laboratories from 13 countries participated in the study using a variety of commercial and in-house quantitative and qualitative assays. The data presented here indicate that, upon freeze-drying, there is no significant loss in potency for the candidate HPV-18 DNA and a slight loss in potency for the candidate HPV 16 DNA; although this is likely not scientifically relevant when assay precision is considered. In general, the individual laboratory mean estimates for each study sample were grouped +/- approximately 2 log(10) around the theoretical HPV DNA concentration of the reconstituted ampoule (1 x 10(7) HPV genome equivalents/mL). The agreement between laboratories is improved when potencies are made relative to the candidate international standards, demonstrating their utility in harmonizing amplification and detection steps of HPV-16 and -18 DNA assays. Degradation studies indicate that the candidate international standards are extremely stable and suitable for long-term use. Based on these findings, the candidate standards were established as the 1st WHO international standards for HPV-16 DNA and HPV-18 DNA, each with a potency of 5 x 10(6) international units (IU) per ampoule or 1 x 10(7) IU mL(-1) when reconstituted as directed. PMID- 19904757 TI - Parent of origin effects on age at colorectal cancer diagnosis. AB - Genomic imprinting refers to a parent-of-origin specific effect on gene expression. At least 1% of genes in the human genome are modulated in this manner. We sought evidence for genomic imprinting in colorectal cancer by studying the ages at diagnosis in the offspring of 2,061 parent-child pairs in which both parent and child were affected by nonsyndromic colorectal cancer. Families were ascertained through the colon Cancer Family Registry [http://epi.grants.cancer.gov/CFR/] from both population-based and clinic-based sources. We found that the affected offspring of affected fathers were on average younger than offspring of affected mothers (55.8 vs. 53.7 years; p = 0.0003), but when divided into sons and daughters, this difference was driven entirely by younger age at diagnosis in daughters of affected fathers compared to sons (52.3 years vs. 55.1 years; p = 0.0004). A younger age at diagnosis in affected daughters of affected fathers was also observable in various subsets including families that met Amsterdam II Criteria, families that did not meet Amsterdam Criteria, and in families with documented normal DNA mismatch repair in tumors. Imprinting effects are not expected to be affected by the sex of the offspring. Possible explanations for these unexpected findings include: (i) an imprinted gene on the pseudoautosomal regions of the X chromosome; (ii) an imprinted autosomal gene that affects a sex-specific pathway; or (iii) an X-linked gene unmasked because of colonic tissue-specific preferential inactivation of the maternal X chromosome. PMID- 19904758 TI - High CCND1 amplification identifies a group of poor prognosis women with estrogen receptor positive breast cancer. AB - CCND1 encodes for the cyclin D1 protein involved in G1/S cell cycle transition. In breast cancer the mechanism of CCND1 amplification, relationship between cyclin D1 protein expression and the key clinical markers estrogen receptor (ER) and HER2 requires elucidation. Tissue microarrays of primary invasive breast cancer from 93 women were evaluated for CCND1 amplification by fluorescent in situ hybridization and cyclin D1 protein overexpression by immunohistochemistry. CCND1 amplification was identified in 27/93 (30%) cancers and 59/93 (63%) cancers had overexpression of cyclin D1. CCND1 amplification was significantly associated with cyclin D1 protein overexpression (p < 0.001; Fisher's exact test) and both CCND1 amplification and cyclin D1 protein expression with oestrogen receptor (ER) expression (p = 0.003 and p < 0.001; Fishers exact test). Neither CCND1 amplification nor cyclinD1 expression was associated with tumor size, pathological node status or HER2 amplification, but high CCND1 amplification (Copy Number Gain (CNG) > or = 8) was associated with high tumor grade (p = 0.005; chi square 7.915, 2 df) and worse prognosis by Nottingham Prognostic Index (p = 0.001; 2 sample t-test). High CCND1 amplification (CNG > or = 8) may identify a subset of patients with poor prognosis ER-positive breast cancers who should be considered for additional therapy. PMID- 19904759 TI - Dietary flaxseed lignan or oil combined with tamoxifen treatment affects MCF-7 tumor growth through estrogen receptor- and growth factor-signaling pathways. AB - This study aimed to elucidate which component of flaxseed, i.e. secoisolariciresinol diglucoside (SDG) lignan or flaxseed oil (FO), makes tamoxifen (TAM) more effective in reducing growth of established estrogen receptor positive breast tumors (MCF-7) at low circulating estrogen levels, and potential mechanisms of action. In a 2 x 2 factorial design, ovariectomized athymic mice with established tumors were treated for 8 wk with TAM together with basal diet (control), or basal diet supplemented with SDG (1 g/kg diet), FO (38.5 g/kg diet), or combined SDG and FO. SDG and FO were at levels in 10% flaxseed diet. Palpable tumors were monitored and after animal sacrifice, analyzed for cell proliferation, apoptosis, ER-mediated (ER-alpha, ER-beta, trefoil factor 1, cyclin D1, progesterone receptor, AIBI), growth factor-mediated (epidermal growth factor receptor, human epidermal growth factor receptor-2, insulin-like growth factor receptor-1, phosphorylated mitogen activated protein kinase, PAKT, BCL2) signaling pathways and angiogenesis (vascular endothelial growth factor). All treatments reduced the growth of TAM-treated tumors by reducing cell proliferation, expression of genes, and proteins involved in the ER- and growth factor-mediated signaling pathways with FO having the greatest effect in increasing apoptosis compared with TAM treatment alone. SDG and FO reduced the growth of TAM-treated tumors but FO was more effective. The mechanisms involve both the ER- and growth factor-signaling pathways. PMID- 19904760 TI - Garlic components inhibit angiotensin II-induced cell-cycle progression and migration: Involvement of cell-cycle inhibitor p27(Kip1) and mitogen-activated protein kinase. AB - Garlic has been used for prevention and treatment of hypertension; however, the molecular mechanisms of garlic's effects remain to be elucidated. In this study, the mechanisms of the in vitro effect of organosulphur compounds derived from garlic on growth and migration of cultured aortic smooth muscle cells isolated from spontaneously hypertensive rats were investigated. We demonstrated that allyl methyl sulphide (AMS) and diallyl sulphide (DAS) inhibited aortic smooth muscle cell angiotensin II-stimulated cell-cycle progression and migration. Neither cell viability nor annexin-V-binding analysis revealed cytotoxic effects of both organosulphur compounds at the used concentrations. Instead, their inhibitory effects were associated to the prevention of the cell-cycle inhibitor p27(Kip1) (p27) downregulation and the reduction of extracellular signal regulated kinase 1/2 phosphorylation. When we assessed the antioxidant activity of AMS and DAS, we found that both organosulphur compounds inhibited angiotensin II-reactive oxygen species generation. Our findings show that AMS and DAS, compounds derivate from garlic, could be effective antioxidants targeted at the arterial remodelling seen in hypertension. PMID- 19904761 TI - Effect of sulforaphane on glutathione-adduct formation and on glutathione_S_transferase-dependent detoxification of acrylamide in Caco-2 cells. AB - The toxicity of dietary acrylamide (AA) depends on its biotransformation pathways, in which phase I cytochrome P-450 enzymes transform AA into glycidamide. The phase II enzyme glutathione_S_transferase (GST) catalyses the conjugation of AA with glutathione (GSH). GST induction by phytochemicals like sulforaphane (SFN) plays a role in chemoprevention. Here, the effect of SFN on the detoxification of AA through GSH conjugation was studied in Caco-2 cells. GSH adducts with AA and SFN were synthesized, identified by NMR and quantified by LC MS/MS. Caco-2 cells were treated with either 2.5 mM AA, 10 microM SFN or the combination of both for 24 h. Concentrations of GSH conjugates (GSH-AA, GSH-SFN, SFN-GSH-AA), AA and SFN were analysed by LC-MS/MS. GSH contents and GST activity were determined photometrically. GST activity was increased after treatment of the cells with SFN (38+/-6%, p< or =0.05) or AA (25+/-4%, p< or =0.05). GSH concentrations decreased after all treatments. Quantitative data of GSH adduct formation showed that the reaction between GSH and SFN is favoured over that between GSH and AA. The data suggest that SFN might impair the GSH-dependent detoxification of AA by SFN-GSH adduct formation and, thus, lower the GSH concentrations available for its reaction with AA. PMID- 19904762 TI - Perspectives and challenges of emerging single-molecule DNA sequencing technologies. AB - The growing demand for analysis of the genomes of many species and cancers, for understanding the role of genetic variation among individuals in disease, and with the ultimate goal of deciphering individual human genomes has led to the development of non-Sanger reaction-based technologies towards rapid and inexpensive DNA sequencing. Recent advancements in new DNA sequencing technologies are changing the scientific horizon by dramatically accelerating biological and biomedical research and promising an era of personalized medicine for improved human health. Two single-molecule sequencing technologies based on fluorescence detection are already in a working state. The newly launched and emerging single-molecule DNA sequencing approaches are reviewed here. The current challenges of these technologies and potential methods of overcoming these challenges are critically discussed. Further research and development of single molecule sequencing will allow researchers to gather nearly error-free genomic data in a timely and inexpensive manner. PMID- 19904763 TI - Multifunctional silver-embedded magnetic nanoparticles as SERS nanoprobes and their applications. AB - In this study, surface-enhanced Raman spectroscopy (SERS)-encoded magnetic nanoparticles (NPs) are prepared and utilized as a multifunctional tagging material for cancer-cell targeting and separation. First, silver-embedded magnetic NPs are prepared, composed of an 18-nm magnetic core and a 16-nm-thick silica shell with silver NPs formed on the surface. After simple aromatic compounds are adsorbed on the silver-embedded magnetic NPs, they are coated with silica to provide them with chemical and physical stability. The resulting silica encapsulated magnetic NPs (M-SERS dots) produce strong SERS signals and have magnetic properties. In a model application as a tagging material, the M-SERS dots are successfully utilized for targeting breast-cancer cells (SKBR3) and floating leukemia cells (SP2/O). The targeted cancer cells can be easily separated from the untargeted cells using an external magnetic field. The separated targeted cancer cells exhibit a Raman signal originating from the M SERS dots. This system proves to be an efficient tool for separating targeted cells. Additionally, the magnetic-field-induced hot spots, which can provide a 1000-times-stronger SERS intensity due to aggregation of the NPs, are studied. PMID- 19904764 TI - Size-selective effects on fullerene adsorption by nanoporous molecular networks. AB - A new hierarchical self-assembling molecular template, which can size-selectively immobilize fullerene molecules, is reported. The molecular template is fabricated from 1,3,5-tris(10-carboxydecyloxy)benzene (TCDB) and triangle-shaped macrocycles. It is observed that the two-dimensional hydrogen-bonded achiral TCDB network affected by the 3NN-Macrocycle becomes a chiral network. Host and guest molecules both form chiral arrangements with hexagonal empty pores. In addition, fullerenes and other molecules such as coronene can be entrapped in the empty pores or on the 3NN-Macrocycle molecules. The adsorption constant (K) is estimated, from which it is concluded that the different filling behaviors of the fullerenes are associated with the different sizes of the guest species. This method provides a facile approach to molecularly designed surfaces and the study of fullerene molecular arrays on the single-molecule level. PMID- 19904765 TI - Proteolytic fluorescent signal amplification on gold nanoparticles for a highly sensitive and rapid protease assay. AB - A new strategy for highly sensitive and rapid protease assay is developed by mediating proteolytic formation of oligonucleotide duplexes and using the duplexes for signal amplification. In the presence of matrix metalloprotease-2 (MMP-2), fragmentation of the intact DNA-peptide on gold nanoparticles (GNP) by hydrolytic cleavage of a peptide bond within the substrate allows diffusion of DNA away from the GNP and the formation of a DNA/RNA heteroduplex, leading to digestion of RNA by RNase H. Because of the high quenching efficacy of GNP to the fluorophore in RNA and multiple digestions of the RNA, the fluorescence signal recovery is amplified. This method permits the assessment of the activity of MMP 2 at concentrations as low as 10 pM within 4 h. Compared with the reported protease nanosensors using quantum dots, GNP, and magnetic nanoparticles with the same peptide sequence, the assay time of this method is sixfold faster and the limit of detection is 100-fold more sensitive. The formulations for proteolytic formations of oligonucleotides duplexes for signal amplification on GNP could lead to the development of more sensitive and rapid protease assay techniques, thus extending the role of proteases as therapeutic targets and disease indicators. PMID- 19904766 TI - Colostrum and bioactive, colostral peptides differentially modulate the innate immune response of intestinal epithelial cells. AB - Characterization and identification of peptides with bioactivity from food have received considerable interest recently since such bioactive components must be adequately documented if they are part of functional food claims. We have characterized peptides from colostrum or those generated by a simulated gastrointestinal digest (GI) and tested them for bioactivity using murine intestinal (mIC(c12)) cells and compared with bioactivity of intact colostrum. The peptides were recovered in the permeate after dialysis. The presence of peptides in the permeate was confirmed by C(18) RP-HPLC, determination of free amino termini and MALDI MS. The bioactivity of the intact colostrum and colostral peptides in the permeate was tested using mIC(c12) cells stimulated in the absence or presence of different bacterial ligands that mediate cellular activation through stimulation of Toll-like receptors (TLR). Whereas intact colostrum generally reduced TLR-mediated signaling, the isolated peptides seemed to either stimulate or reduce the immune response depending on the bacterial ligand used for stimulation. Interestingly, the most potent bioactive peptides originated from nondigested colostrum, which had only been subject to endogenous protease activity. Identified peptides in the nondigested colostrum originated exclusively from the casein fraction of colostrum as shown by MALDI MS/MS identification. Thus, multiple components with different bioactivities towards the innate immune response appear in bovine colostrum. PMID- 19904767 TI - Activating NK-cell receptors co-stimulate CD4(+)CD28(-) T cells in patients with rheumatoid arthritis. AB - Effector T-cell responses can be modulated by competing positive or negative signals transduced by NK-cell receptors (NKR). In the CD4(+) T-cell population, the expression of NKR is primarily found in the CD4(+)CD28(-) T-cell subset, also known as CD28(null) T cells. These T cells are frequently found in rheumatoid arthritis (RA) and other inflammatory disorders, suggesting that signaling through NKR may play a role in the autoimmune reaction. Here we aimed to dissect the phenotype and function of NKR-expressing CD4(+)CD28(-) T cells in patients with RA. By analyzing a broad array of NKR on CD4(+)CD28(-) T cells we found a significant expression of the co-activating receptors 2B4 (CD244), DNAM-1 (CD226), and CRACC. Pair-wise ligations of 2B4 with DNAM-1 and/or NKG2D lead to increased effector functions of primary CD4(+)CD28(-) T cells to suboptimal levels of anti-CD3 stimulation. Using multi-parameter flow cytometry, we demonstrate that such co-ligation led to an increased magnitude in overall responsiveness without changing qualitative aspects of the response. Altogether these results demonstrate a pattern of additive effects in NKR-mediated functional modulation of CD4(+)CD28(-) T cells in RA. This may have consequences for the inflammatory responses imposed by these cells, thus influencing disease manifestations. PMID- 19904768 TI - MYD88-dependent and -independent activation of TREM-1 via specific TLR ligands. AB - Triggering receptor expressed on myeloid cells (TREM)-1 plays an important role in myeloid cell-activated inflammatory responses. Although TLR ligands such as LPS and lipoteichoic acid have been shown to upregulate TREM-1 expression in macrophage and neutrophils, the role of specific TLR in inducing the expression of TREM-1 remains unclear. In this study, we investigated whether the presence of TLR is necessary for the expression of TREM-1. We show that BM-derived macrophages from TLR4 and TLR2 KO mice failed to induce expression of TREM-1 message and protein in response to their specific ligands. Interestingly, the expression of TREM-1 in response to LPS is not altered in myeloid differentiation factor 88 (MyD88) KO macrophages, suggesting that downstream of TLR a MyD88 independent pathway induces the expression of TREM-1. Inhibiting toll/IL-1R domain-containing adaptor-inducing IFN-beta (TRIF) expression by siRNA decreased TREM-1 expression in response to LPS, suggesting that the expression of TREM-1 in response to LPS was mediated by the TRIF signaling pathway. On the other hand, the expression of TREM-1 in response to lipoteichoic acid is dependent on MyD88 expression. These data indicate that the expression of TREM-1 in response to TLR ligands occurs secondary to downstream signaling events and that the presence of TLR is necessary for the expression of TREM-1 in response to their specific ligands. However, the downstream signaling required for the expression of TREM-1 is dependent on the stimulus and the surface receptor through which the signaling is initiated. PMID- 19904769 TI - Unaltered negative selection and Treg development of self-reactive thymocytes in TCR transgenic Fyn-deficient mice. AB - The tyrosine kinase Fyn has been implicated as playing an important role in the generation of both stimulatory and inhibitory signaling events induced by TCR engagement. To assess the role of Fyn for antigen-driven negative selection and Treg development, which are both dependent on the strength and nature of TCR signaling, we generated mice that co-express the transgenes for OVA and the OT-II TCR, which recognizes a peptide from OVA. In mice expressing both transgenes, negative selection, Treg development in the thymus, and the number of Treg in the periphery were each unaffected by ablation of Fyn. Moreover, fyn(-/-) Treg were functional, as assessed in vitro. We further tested the role of Fyn for the adaptor function of c-Cbl, using mice containing a point mutation in c-Cbl that abolishes its E3 ubiquitin ligase function but maintains its adaptor function. The functional and signaling properties of this mutant c-Cbl were unaltered in fyn(-/-) thymocytes. Combined, these data indicate that Fyn was not required for the induction of central tolerance by negative selection, the adaptor protein role of c-Cbl, or the normal development and function of Treg. PMID- 19904770 TI - The Tregs' world according to GARP. AB - Naturally occurring CD4+CD25(high) regulatory T cells (nTreg) are essential for maintaining tolerance. FOXP3 has been established as a molecular marker of nTreg; however, FOXP3 cannot be used as a reliable marker for bona fide human nTreg since effector T cells also up-regulate FOXP3 expression upon activation. Despite the important function of nTreg, the underlying molecular mechanisms of nTreg mediated suppression are far from defined. Previous studies have demonstrated that the TGF-beta latency-associated peptide (LAP) is expressed on the surface of nTreg, and that immunosuppression can be mediated by membrane TGF-beta; however, it remains unknown how LAP is bound to nTreg and what is the functional significance of its selective expression on activated nTreg. The nTreg's world may now change according to GARP, an orphan toll-like receptor composed of leucine-rich repeats. In this issue of the European Journal of Immunology, a study provides further demonstration that GARP is selectively expressed only in activated human nTreg and nTreg cell clones but not in activated effector T cells, confirming GARP as a bona fide nTreg marker. In addition, GARP binds directly to LAP; yet, GARP over-expression is insufficient to induce modification of latent TGF-beta into active TGF-beta further clarifying its role in nTreg mediated suppression. PMID- 19904771 TI - The loss of phenol sulfotransferase 1 in hepatocellular carcinogenesis. AB - Biomarkers for the detection of early hepatocellular carcinoma (HCC) are urgently needed. To identify biomarkers of HCC, we performed a comparative proteomics analysis, based on 2-DE of HCC tissues and surrounding non-tumor tissues. Six xenobiotic enzymes were significantly down-regulated in the HCC tissue. Among these, phenol sulfotransferase (SULT1A1) was confirmed by Western blot analysis in 105 HCC patients. SULT1A1 showed a significant decrease in 98.1% of the HCC tissues, with 88.6% sensitivity and 66.7% specificity for the detection of HCC. Immunohistochemistry for SULT1A1 was performed and compared with glypican-3, which is a well-known marker of HCC. The results showed down-regulation of SULT1A1 and up-regulation of glypican-3 in 52.6 and 71.9% of the HCCs, and the use of both markers improved the sensitivity up to 78.9%. Moreover, SULT1A1 was useful in differentiating early HCC from benign dysplastic nodules. Clinically, the down-regulation of SULT1A1 was closely associated with an advanced International Union Against Cancer stage and high levels of serum alpha fetoprotein. In conclusion, the results of this study demonstrate that the loss of SULT1A1 appears to be a characteristic molecular signature of HCC. SULT1A1 might be a useful biomarker for the detection of early HCC and help predict the clinical outcome of patients with HCC. PMID- 19904772 TI - Bayesian spatial modeling of disease risk in relation to multivariate environmental risk fields. AB - The relationship between exposure to environmental chemicals during pregnancy and early childhood development is an important issue that has a spatial risk component. In this context, we have examined mental retardation and developmental delay (MRDD) outcome measures for children in a Medicaid population in South Carolina and sampled measures of soil chemistry (e.g. As, Hg, etc.) on a network of sites that are misaligned to the outcome residential addresses during pregnancy. The true chemical concentration at the residential addresses is not observed directly and must be interpolated from soil samples. In this study, we have developed a Bayesian joint model that interpolates soil chemical fields and estimates the associated MRDD risk simultaneously. Having multiple spatial fields to interpolate, we have considered a low-rank Kriging method for the interpolation that requires less computation than the Bayesian Kriging. We performed a sensitivity analysis for a bivariate smoothing, changing the number of knots and the smoothing parameter. These analyses show that a low-rank Kriging method can be used as an alternative to a full-rank Kriging, reducing the computational burden. However, the number of knots for the low-rank Kriging model needs to be selected with caution as a bivariate surface estimation can be sensitive to the choice of the number of knots. PMID- 19904773 TI - Optimal combination of estimating equations in the analysis of multilevel nested correlated data. AB - Multilevel nested, correlated data often arise in biomedical research. Examples include teeth nested within quadrants in a mouth or students nested within classrooms in schools. In some settings, cluster sizes may be large relative to the number of independent clusters and the degree of correlation may vary across clusters. When cluster sizes are large, fitting marginal regression models using Generalized Estimating Equations with flexible correlation structures that reflect the nested structure may fail to converge and result in unstable covariance estimates. Also, the use of patterned, nested working correlation structures may not be efficient when correlation varies across clusters. This paper describes a flexible marginal regression modeling approach based on an optimal combination of estimating equations. Particular within-cluster and between-cluster data contrasts are used without specification of the working covariance structure and without estimation of covariance parameters. The method involves estimation of the covariance matrix only for the vector of component estimating equations (which is typically of small dimension) rather than the covariance matrix of the observations within a cluster (which may be of large dimension). In settings where the number of clusters is large relative to the cluster size, the method is stable and is highly efficient, while maintaining appropriate coverage levels. Performance of the method is investigated with simulation studies and an application to a periodontal study. PMID- 19904774 TI - A bifunctional Pd/MgO solid catalyst for the one-pot selective N-monoalkylation of amines with alcohols. AB - It has been found that a bifunctional metal Pd/base (MgO) catalyst performs the selective monoalkylation of amines with alcohols. The reaction goes through a series of consecutive steps in a cascade mode that involves: 1) the abstraction of hydrogen from the alcohol that produces the metal hydride and the carbonyl compound; 2) condensation of the carbonyl with the amine to give an imine, and 3) hydrogenation of the imine with the surface hydrogen atoms from the metal hydride. Based on isotopic and spectroscopic studies and on the rate of each elementary step, a global reaction mechanism has been proposed. The controlling step of the process is the hydride transfer from the metal to the imine. By changing the crystallite size of the Pd, it is demonstrated that this is a structure-sensitive reaction, whereas the competing processes that lead to subproducts are not. On these bases, a highly selective catalyst has been obtained with Pd crystallite size below 2.5 nm in diameter. The high efficiency of the catalytic system has allowed us to extend the process to the one-pot synthesis of piperazines. PMID- 19904775 TI - Mechanistic insights into the substrate-controlled stereochemistry of glycals in one-pot rhodium-catalyzed aziridination and aziridine ring opening. AB - We carried out a principle study on the reaction mechanism of rhodium-catalyzed intramolecular aziridination and aziridine ring opening at a sugar template. A sulfamate ester group was introduced at different positions of glycal to act as a nitrene source and, moreover, to allow the study of the relative reactivity of the nitrene transfer from different sites of the glycal molecule. The structural optimization of each intermediate along the reaction pathway was extensively done by using BPW91 functional. The crucial step in the reaction is the Rh-catalyzed nitrene transfer to the double bond of the glycal. We found that the reaction could proceed in a stepwise manner, whereby the N atom initially induced a single bond formation with C1 on the triplet surface or in a single step through intersystem crossing (ISC) of the triplet excited state of the rhodium-nitrene transition state to the singlet ground state of the aziridine complexes. The relative reactivity for the conversion of the nitrene species to the aziridine obtained from the computed potential energy surface (PES) agrees well with the reaction time gained from experimental observation. The aziridine ring opening is a spontaneous process because the energy barrier for the formation of the transition state is very small and disappears in the solution calculations. The regio- and stereoselectivity of the reaction product is controlled by the electronic property of the anomeric carbon as well as the facial preference for the nitrene insertion, and the nucleophilic addition. PMID- 19904776 TI - Palladium-catalyzed hydroalkynylation of alkylidenecyclopropanes. PMID- 19904778 TI - Stereoselective control by face-to-face versus edge-to-face aromatic interactions: the case of C(3)-Ti(IV) amino trialkolate sulfoxidation catalysts. AB - The stereoselective oxidation of differently functionalised benzyl phenyl sulfides has been examined by using enantiopure Ti(IV) trialkanolamine complexes. These complexes efficiently catalyse the sulfoxidation with good stereoselectivities. The data highlight the contribution to the stereoselectivity of steric effects and non-covalent pi-pi interactions between the aromatic rings of the Ti(IV) complex and those pertaining to the substrates. Enantiomeric excesses have been correlated with the electrostatic potential surfaces (EPS) of the reacting sulfides. The overall study leads to a mechanistic interpretation that explains the stereoselectivity of the system and dissects the role of aromatic and steric interactions in the stereoselective process. PMID- 19904777 TI - The palladium-catalyzed aerobic kinetic resolution of secondary alcohols: reaction development, scope, and applications. AB - The first palladium-catalyzed enantioselective oxidation of secondary alcohols has been developed, utilizing the readily available diamine (-)-sparteine as a chiral ligand and molecular oxygen as the stoichiometric oxidant. Mechanistic insights regarding the role of the base and hydrogen-bond donors have resulted in several improvements to the original system. Namely, addition of cesium carbonate and tert-butyl alcohol greatly enhances reaction rates, promoting rapid resolutions. The use of chloroform as solvent allows the use of ambient air as the terminal oxidant at 23 degrees C, resulting in enhanced catalyst selectivity. These improved reaction conditions have permitted the successful kinetic resolution of benzylic, allylic, and cyclopropyl secondary alcohols to high enantiomeric excess with good-to-excellent selectivity factors. This catalyst system has also been applied to the desymmetrization of meso-diols, providing high yields of enantioenriched hydroxyketones. PMID- 19904779 TI - Structure-reactivity relationships in negishi cross-coupling reactions. AB - Competition experiments have been performed to determine the relative reactivities of substituted bromobenzenes and of different arylzinc reagents in the [Pd(PPh(3))(4)]-catalyzed Negishi cross-coupling reaction in THF at 25 degrees C. The cross-coupling reactions are accelerated by electron acceptors in the bromobenzenes, the effect of which increases in the order ortho or =2 cycles of systemic chemotherapy. Thirty (55.6%) patients had a disease control rate noted as a complete response, partial response, or stable disease. After a median follow-up of 24 months, the median progression-free survival (PFS) and overall survival were determined to be 7.6 months (95% confidence interval [CI], 4-11) and 56 months (95% CI, 36-not applicable), respectively. CONCLUSIONS: Systemic chemotherapy has a role in appendiceal neoplasm patients who are suboptimal candidates for cytoreductive surgery. The intermediate PFS indicates the challenges that exist for appendiceal neoplasm patients in this setting. Prospective randomized trials including systemic chemotherapy are needed to provide further insight into this malignancy, for which no standard exists. PMID- 19904806 TI - Empirically supported religious and spiritual therapies. AB - This article evaluated the efficacy status of religious and spiritual (R/S) therapies for mental health problems, including treatments for depression, anxiety, unforgiveness, eating disorders, schizophrenia, alcoholism, anger, and marital issues. Religions represented included Christianity, Islam, Taoism, and Buddhism. Some studies incorporated a generic spirituality. Several R/S therapies were found to be helpful for clients, supporting the further use and research on these therapies. There was limited evidence that R/S therapies outperformed established secular therapies, thus the decision to use an R/S therapy may be an issue of client preference and therapist comfort. PMID- 19904807 TI - In reference to Transcricothyroid electromyographic monitoring of the recurrent laryngeal nerve. PMID- 19904808 TI - Concerns about substandard training for prescription privileges for psychologists. AB - This Commentary on the Muse and McGrath study (this issue, pp. 96-103) refutes its conclusion that the amount of training for prescription privileges for psychologists (RxP) is equal to or greater than that for psychiatric nurse practitioners and physicians. First, the sample failed to include only training programs for nurses and psychologists that lead to independent prescribing. Second, training was defined by an arbitrary, nonvalidated list of "key content areas" that excluded much of the standard medical curricula for nurses and physicians to prescribe. Third, the levels of training for which the "key content areas" were assessed omitted undergraduate prerequisites, apprenticeships, supervised practice, and residencies that are a standard part of the nursing and physician programs studied. RxP training remains substandard. PMID- 19904809 TI - Rumination fosters indecision in dysphoria. AB - This study investigated the effects of rumination on indecision, assessed as high levels of perceived decision difficulty, low confidence in a decision, and decision latency. Dysphoric and nondysphoric participants were assigned to either a rumination or a distraction induction. Subsequently, they made four decisions with alleged real-life consequences. As predicted, rumination exhibited a negative effect on dysphoric participants' decision-making process. They experienced the decisions as more difficult and had less confidence in their choices. No effects emerged on the measure of decision time. Mediation analyses revealed that increased difficulty of the decisions was due to self-focused thinking as a cognitive consequence of rumination, while reduced confidence in the decisions was partly mediated by negative affect that resulted from rumination. The finding that rumination affects the important life domain of decision making by fostering indecision in dysphoric individuals is a central extension of previous studies on rumination's consequences. In addition, these results provide insight into the depressive symptom of indecisiveness by revealing its underlying mechanisms. PMID- 19904810 TI - The impact of dichotomization in longitudinal data analysis: a simulation study. AB - In this paper, a simulation study is conducted to systematically investigate the impact of dichotomizing longitudinal continuous outcome variables under various types of missing data mechanisms. Generalized linear models (GLM) with standard generalized estimating equations (GEE) are widely used for longitudinal outcome analysis, but these semi-parametric approaches are only valid under missing data completely at random (MCAR). Alternatively, weighted GEE (WGEE) and multiple imputation GEE (MI-GEE) were developed to ensure validity under missing at random (MAR). Using a simulation study, the performance of standard GEE, WGEE and MI-GEE on incomplete longitudinal dichotomized outcome analysis is evaluated. For comparisons, likelihood-based linear mixed effects models (LMM) are used for incomplete longitudinal original continuous outcome analysis. Focusing on dichotomized outcome analysis, MI-GEE with original continuous missing data imputation procedure provides well controlled test sizes and more stable power estimates compared with any other GEE-based approaches. It is also shown that dichotomizing longitudinal continuous outcome will result in substantial loss of power compared with LMM. PMID- 19904812 TI - Diabetes-related changes in auditory brainstem responses. AB - OBJECTIVES/HYPOTHESIS: Determine effects on auditory brainstem response (ABR) of diabetes mellitus (DM) severity. STUDY DESIGN: A cross-sectional study investigating DM severity and ABR in military Veteran subjects with (166) and without (138) DM and with no more than moderate hearing loss. METHODS: Subjects were classified by three age tertiles (<50, 50-56, and 57+). DM severity was classified as insulin-dependent (IDDM), non-insulin-dependent (NIDDM), or no DM. Other DM measures included serum glucose, HbA1c, and several DM-related complications. ABR measures included wave I, III, and V latencies; I-III, III-V, and I-V latency intervals; and wave V amplitude; for each ear at three repetition rates (11, 51, and 71 clicks/second), and both polarities. Outcomes were stratified by age tertile and adjusted for pure tone threshold at 3 kHz. Repeated measures multivariate analysis of covariance modeled the ABR response at each repetition rate for DM severity (main effect) and hearing at 3 kHz (covariate). Modeled contrasts between ABR variables in subjects with and without DM were examined. RESULTS: Significant differences existed between no DM and IDDM groups in the younger tertile only. Adjusting for threshold at 3 kHz had minimal effect. Self-reported noise exposure was not related to ABR differences, but HbA1c and poor circulation were. CONCLUSIONS: IDDM is associated with an increased wave V latency, wave I-V interval, and reduced wave V amplitude among Veterans under 50 years. Results were related to several DM complications. PMID- 19904813 TI - Steroid receptor phosphorylation: Assigning function to site-specific phosphorylation. AB - Steroid receptors (SRs) are hormone-activated transcription factors important for a wide variety of cellular functions. Post-translational modifications of SRs, including phosphorylation, ubiquitination, acetylation, and sumoylation regulate their expression and function. The remarkable number of phosphorylation sites in these receptors and the wide variety of kinases shown to modulate phosphorylation influence the integration between cell-signaling pathways and SR action. These phosphorylation sites have been identified in all of the functional domains with the majority being located within the amino-terminal portions of the receptors. The regulation of function is receptor specific, site specific, and often dependent on the cellular context. Numerous roles for site-specific phosphorylation have been elucidated including sensitivity of hormone response, DNA binding, expression, stability, subcellular localization, dimerization, and protein-protein interactions that can determine the regulation of specific target genes. This review summarizes the current knowledge regarding receptor site specific phosphorylation and regulation of function. As functional assays become more sophisticated, it is likely that additional roles for phosphorylation in receptor function will be identified. PMID- 19904815 TI - Characterization of skin inflammation induced by repeated exposure of toluene, xylene, and formaldehyde in mice. AB - Volatile organic compounds (VOCs) are considered the main cause of sick building syndrome and they are likely to irritate the skin, eyes, and mucous membrane; however, the toxic threshold and the mechanisms of cutaneous reaction induced by long-time VOC exposure have not been clarified. In the present study, we investigated the effect of repeated painting of VOCs onto mouse skin. Various concentrations of toluene, xylene, and formaldehyde (FA) were applied once a week for 5 weeks. While FA solution (2-10%) induced remarkable ear swelling and caused evident infiltration of inflammatory cells, high concentrations of toluene and xylene (50 or 100%) evoked mild ear swelling and marginal inflammatory cell invasion. In addition, FA exposure markedly increased the expression of interleukin-4 (IL-4), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and transient receptor potential vanilloid-1 (TRPV-1) mRNAs in the ears and IL-4 and NT-3 mRNAs in the cervical lymph nodes. Furthermore, capsazepine, a TRPV-1 antagonist, significantly suppressed ear swelling caused by repeated painting of 5% FA. These findings demonstrate that FA has more potent irritancy against skin than toluene or xylene and suggest that the Th2 response, neurotrophins and TRPV-1 play important roles in FA-induced skin inflammation. PMID- 19904816 TI - Central and peripheral oxygen transmissibility thresholds to avoid corneal swelling during open eye soft contact lens wear. AB - This study was designed to derive central and peripheral oxygen transmissibility (Dk/t) thresholds for soft contact lenses to avoid hypoxia-induced corneal swelling (increased corneal thickness) during open eye wear. Central and peripheral corneal thicknesses were measured in a masked and randomized fashion for the left eye of each of seven subjects before and after 3 h of afternoon wear of five conventional hydrogel and silicone hydrogel contact lens types offering a range of Dk/t from 2.4 units to 115.3 units. Curve fitting for plots of change in corneal thickness versus central and peripheral Dk/t found threshold values of 19.8 and 32.6 units to avoid corneal swelling during open eye contact lens wear for a typical wearer. Although some conventional hydrogel soft lenses are able to achieve this criterion for either central or peripheral lens areas (depending on lens power), in general, no conventional hydrogel soft lenses meet both the central and peripheral thresholds. Silicone hydrogel contact lenses typically meet both the central and peripheral thresholds and use of these lenses therefore avoids swelling in all regions of the cornea. PMID- 19904817 TI - Controlled release of tetracycline from biodegradable beta-tricalcium phosphate composites. AB - For use in the prevention of bone infections, a novel controlled release system composed of beta-tricalcium phosphate (TCP) granules with biodegradable coatings incorporating the antibiotic drug tetracycline (TC) was developed. Six formulations using poly(D,L-lactide) and poly(D,L-lactide-co-glycolide) as coating materials to incorporate TC were prepared and tested in vitro and in vivo. Release of TC from TCP composites was dependent on the biodegradability of the used polymers and on physical-chemical interactions of TC with the polymer materials. Three characteristic release profiles were obtained: slow release lasting up to 67 days, intermediate release with 60% of the total dose released up to day 20, and fast release with a high initial burst and 90% of TC released within 4 days. Even though TC decomposition products had formed during in vitro release, no cytotoxic effects on osteoblast-like cells were observed. The biological activity of TC after incorporation into PL(G)A films was confirmed using a TC-repressible promoter system in genetically engineered Chinese Hamster Ovary (CHO) cells. TC-loaded TCP composites implanted in ovine cancellous bone defects showed good biocompatibility and new bone formation in the histological evaluation. No differences in the cellular reactions were seen between antibiotic loaded composites and the control group. These experimental results indicate the potential of coated TCP composites to be used as local carrier system for controlled TC delivery with different release kinetics and good in vitro and in vivo biocompatibility. PMID- 19904818 TI - In vivo antibacterial and silver-releasing properties of novel thermal sprayed silver-containing hydroxyapatite coating. AB - One of the serious postoperative complications associated with joint replacement is bacterial infection. In addressing this problem, we have previously described the development of a novel thermal spraying technology combining silver (Ag) showing antibacterial activity with hydroxyapatite (HA) displaying good biocompatibility and osteoconductivity, and reported the in vitro properties. This study evaluated serum Ag ion concentrations and antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) using a subcutaneous rat model. HA loaded with 3 wt % of silver oxide (Ag-HA) and plain HA were sprayed on the surface of titanium disks. Ag-HA- or HA-coated samples were implanted into the back subcutaneous pockets of male Sprague-Dawley rats. Mean serum Ag ion concentration in the Ag-HA group increased to more than 50 ppb by 48 h after implantation, then decreased gradually to baseline levels. Mean (+/- standard error of the mean) number of viable MRSA on HA coating was (1.5 +/- 0.5) x 10(5), which is significantly more than the (1.1 +/- 0.4) x 10(4) on Ag-HA coating (p < 0.001). Ag-HA coating offers good abilities to release Ag ions and kill MRSA in vivo. PMID- 19904819 TI - IL-6 adsorption dynamics in hemoadsorption beads studied using confocal laser scanning microscopy. AB - Sepsis is characterized by a systemic inflammatory response caused by infection, and can result in organ failure and death. Removal of inflammatory mediators such as cytokines from the circulating blood is a promising treatment for severe sepsis. We are developing an extracorporeal hemoadsorption device to remove cytokines from the blood using biocompatible, polymer sorbent beads. In this study, we used confocal laser scanning microscopy (CLSM) to directly examine adsorption dynamics of a cytokine (IL-6) within hemoadsorption beads. Fluorescently labeled IL-6 was incubated with sorbent particles, and CLSM was used to quantify spatial adsorption profiles of IL-6 within the sorbent matrix. IL-6 adsorption was limited to the outer 15 microm of the sorbent particle over a relevant clinical time period, and intraparticle adsorption dynamics was modeled using classical adsorption/diffusion mechanisms. A single model parameter, alpha = q(max) K/D, was estimated by fitting CLSM intensity profiles to our mathematical model, where q(max) and K are Langmuir adsorption isotherm parameters, and D is the effective diffusion coefficient of IL-6 within the sorbent matrix. Given the large diameter of our sorbent beads (450 microm), less than 20% of available sorbent surface area participates in cytokine adsorption. Development of smaller beads may accelerate cytokine adsorption by maximizing available surface area per bead mass. PMID- 19904814 TI - Current status of mucins in the diagnosis and therapy of cancer. AB - Mucins are the most abundant high molecular weight glycoproteins in mucus. Their nature and glycosylation content dictates the biochemical and biophysical properties of viscoelastic secretions, pointing out an important role in diverse biological functions, such as differentiation, cell adhesions, immune responses, and cell signaling. Mucins are expressed in tubular organs by specialized epithelial cells in the body. Their aberrant expression is well documented in a variety of inflammatory or malignant diseases. From a prognosis point of view, their expression and alterations in glycosylation are associated with the development and progression of malignant diseases. Therefore, mucins can be used as valuable markers to distinguish between normal and disease conditions. Indeed, this alteration in glycosylation patterns generates several epitopes in the oligosaccharide side chains that can be used as diagnostic and/or prognostic markers. Furthermore, these characteristic tumor-associated epitopes are extensively used as appropriate immunotargets of malignant epithelial cells. Therefore, in an effort to detect and treat cancer at the earliest stage possible, mucins are analyzed as potential markers of disease for diagnosis, progression, and for therapeutic purposes. In this review, we focused on the current status of the distribution of mucins in normal and pathologic conditions and their clinical use both in cancer diagnosis and therapeutics treatments. PMID- 19904820 TI - Physicochemical characterization of biomaterials commonly used in dentistry as bone substitutes--comparison with human bone. AB - The present work focuses on the physicochemical characterization of selected mineral-based biomaterials that are frequently used in dental applications. The selected materials are commercially available as granules from different biological origins: bovine, porcine, and coralline. Natural and calcined human bone were used for comparison purposes. Besides a classical rationalization of chemical composition and crystallinity, a major emphasis was placed on the measurement of various morphostructural properties such as particle size, porosity, density, and specific surface area. Such properties are crucial to acquiring a full interpretation of the in vivo performance. The studied samples exhibited distinct particle sizes (between 200 and 1000 microm) and shapes. Mercury intrusion revealed not only that the total sample porosity varied considerably (33% for OsteoBiol, 50% for PepGen P-15, and 60% for BioOss) but also that a significant percentage of that porosity corresponded to submicron pores. Biocoral was not analyzed by this technique as it possesses larger pores than those of the porosimeter upper limit. The density values determined for the calcined samples were close to the theoretical values of hydroxyapatite. However, the values for the collagenated samples were lower, in accordance with their lower mineral content. The specific surface areas ranged from less than 1 m(2)/g (Biocoral) up to 60 m(2)/g (BioOss). The chemical and phase composition of most of the samples, the exception being Biocoral (aragonite), were hydroxyapatite based. Nonetheless, the samples exhibited different organic material content as a consequence of the distinct heat treatments that each had received. PMID- 19904821 TI - Synthesis and characterization of functionalized magnetic maghemite nanoparticles with fluorescent probe capabilities for biological applications. AB - Nanoparticles with innovative optical, chemical, and magnetic properties combined in a single nanoparticle may be useful as biosensors, targeting agents, and therapeutic agents in the biomedical field. This study describes new magnetic nanoparticles (MNPs) containing the fluorescent dye rhodamine isothiocyanate (RITC) covalently encapsulated within the nanoparticles. These nanoparticles have been prepared by nucleation followed by controlled growth of iron oxide layers onto iron oxide/gelatin-RITC nuclei. The formed RITC labeled MNPs (R-MNPs) are of narrow size distribution, exhibit the fluorescent spectrum of RITC, yet are more photostable. Because of the covalent encapsulation of RITC within the MNPs no detectable leakage of the fluorescent dye into the aqueous continuous phase was observed. This manuscript also demonstrates that the surface of the R-MNPs retains similar ligand binding efficiency as the equivalent nonfluorescent MNPs. Specific cell labeling was obtained by incubating glia cells with R-MNPs conjugated to glial cell line-derived neurotrophic factor (GDNF) protein. We further showed that the R-MNPs may be used for pH sensing between the pH range of 5 and 9. This feature may enable the use of the R-MNPs as a pH sensor of animal tissues and cell compartments. Thus, these functional narrow size distribution R MNPs with both magnetic and fluorescent properties may provide an important research tool for biological sensing. PMID- 19904822 TI - Osteotomy of distal radius fracture malunion using a fast remodeling bone substitute consisting of calcium sulphate and calcium phosphate. AB - Malunion after a distal radius fracture can be treated with an osteotomy of the distal radius. Autologous iliac crest bone graft is often used to fill the gap, but the procedure is associated with donor site morbidity. In this study a novel fast resorbing biphasic bone substitute consisting of a mixture of calcium sulphate and calcium phosphate is used (Cerament BoneSupport AB, Sweden). Fifteen consecutive patients, with a mean age of 52 (27-71) years were included. All had a malunion after a distal radius fracture and underwent an osteotomy. A fragment specific fixation system, TriMed (TriMed, Valencia, CA), consisting of a Buttress Pin and a Radial Pin Plate were used for fixation and a calcium sulphate and calcium phosphate mixture as bone substitute. The patients were followed for 1 year. Grip strength increased from 61 (28-93)% of the contralateral hand to 85 (58-109)%, p < 0.001. DASH scores decreased from 37 (22-61) to 24 (2-49) p = 0.003. Radiographically all osteotomies healed. An increase of ulnar variance was noted during healing from 1.8 mm immediately postoperatively to 2.6 mm at final follow up. Osteotomy can increase grip strength and decease disability after a malunited fracture. In the present series the bone substitute was replaced by bone, but a minor loss of the achieved radiographic correction was noted in some patients during osteotomy healing. A more rigid fixation may improve the radiographic outcome with this kind of bone substitute. PMID- 19904823 TI - The mechanical and biological properties of an injectable calcium phosphate cement-fibrin glue composite for bone regeneration. AB - Calcium phosphate cement (CPC) that can be injected to form a scaffold in situ has promise for the repair of bone defects. However, its low-strength limits the CPC to non-stress-bearing repairs. Fibrin glue (FG) with good sticking property and biocompatibility is possible used to reinforce the CPC. The objective of this study was to investigate the effects of FG on the mechanical and biological properties of CPC in an injectable CPC-FG composite. The initial setting time of this CPC-FG was delayed compared with the CPC control at different powder/liquid (P/L) mass ratio (p > 0.05). At a P/L of 5, the strength was (38.41 +/- 4.32) MPa for the CPC-FG, much higher than (27.42 +/- 2.85) MPa for the CPC alone (p < 0.05). SEM showed bone marrow stromal cells (BMSCs) with healthy spreading and anchored on the CPC-FG composite. After 14 days, the alkaline phosphatase (ALP) activity was (538 +/- 33) for the BMSCs on the CPC-FG and (517 +/- 27) for the BMSCs on the CPC alone. Both ALPs were higher than the baseline ALP (93 +/- 10) for the undifferentiated BMSCs (p < 0.05). The results demonstrate that this stronger CPC-FG scaffold may be useful for stem cell-based bone regeneration in moderate load-bearing orthopedic applications. PMID- 19904824 TI - A review--micromorphological evidence of degradation in resin-dentin bonds and potential preventional solutions. AB - Resin adhesion to dentin was first achieved in 1982 through mechanical hybridization between resin and collagen fibrils using an adhesive resin containing a functional monomer. Over the last 2 decades, newly developed adhesive resins have attempted to improve the bond strength at least in the first 24 h after bonding. Although much is known about the initial bond strength, learned through morphological analysis, the long-term durability of bonds has not yet been established analytically. However, numerous recent studies have shown chemical biodegradation of resin-dentin bonds under various testing regimes. In general, studies have shown that dentin bond strength decreases over time due to degradation of the resin and the collagen fibrils within the bonds. Furthermore, crystal formation around cured bonding resins has been reported in a number of adhesives after long-term water storage. However, the extent and process of degradation or crystallization is adhesive type specific. This review mainly summarizes the most recent and state of the art work in degradation of the bonding of dental restorative compounds with dentin based on micromorphological data of scanning and transmission electron microscopy. PMID- 19904825 TI - Moisture-activated rheological structuring of nonaqueous poloxamine-poly(acrylic acid) systems designed as novel biomedical implants. AB - This study reports the formulation/characterisation of novel polymeric platforms designed to behave as low-viscosity systems in the nonaqueous state, however, following uptake of aqueous fluids, exhibit rheological structuring and mucoadhesion. The rheological/mechanical and mucoadhesive properties of platforms containing poly(acrylic acid) (PAA, 1%, 3%, 5%, w/w) and poloxamines (Tetronic 904, 901, 704, 701, 304), both in the absence and presence of phosphate buffered saline (PBS, pH 7.4) are described. With the exception of Tetronic 904, all formulations exhibited Newtonian flow in the nonaqueous state, whereas, all aqueous formulations displayed pseudoplastic flow. The consistency and viscoelastic properties were dependent on the concentrations of PAA and PBS and Tetronic grade. PBS significantly increased the consistency, viscoelasticity and mucoadhesion, reaching a maximum at a defined concentration of PBS that was dependent on PAA concentration and Tetronic grade. Formulations containing Tetronic 904 exhibited greatest consistency and elasticity both prior to and after dilution with PBS. Increasing PAA concentration enhanced the mucoadhesive properties. Prolonged drug release of metronidazole was observed from formulations containing 10% (w/w) PBS, 3% and, particularly, 5% (w/w) PAA. It is suggested that the physicochemical properties of formulations containing 3% or 5% (w/w) PAA and Tetronic 904, would render them suitable platforms for administration to body cavities. PMID- 19904826 TI - Influence of pH on transungual passive and iontophoretic transport. AB - The present study investigated the effects of pH on nail permeability and the transport of ions such as sodium (Na) and chloride (Cl) ions endogenous to nail and hydronium and hydroxide ions present at low and high pH, which might compete with drug transport across hydrated nail plate during iontophoresis. Nail hydration and passive transport of water across the nail at pH 1-13 were assessed. Subsequently, passive and iontophoretic transport experiments were conducted using (22)Na and (36)Cl ions under various pH conditions. Nail hydration was independent of pH under moderate pH conditions and increased significantly under extreme pH conditions (pH >11). Likewise, nail permeability for water was pH independent at pH 1-10 and an order of magnitude higher at pH 13. The results of passive and iontophoretic transport of Na and Cl ions are consistent with the permselective property of nail. Interestingly, extremely acidic conditions (e.g., pH 1) altered nail permselectivity with the effect lasting several days at the higher pH conditions. Hydronium and hydroxide ion competition in iontophoretic transport was generally negligible at pH 3-11 was significant at the extreme pH conditions studied. PMID- 19904827 TI - Liposomes with high encapsulation capacity for paclitaxel: Preparation, characterisation and in vivo anticancer effect. AB - Paclitaxel (PTX) is approved for the treatment of ovarian and breast cancer. The commercially available preparation of PTX, Cremophor EL(R) is associated with hypersensitivity reactions in spite of a suitable premedication. In general, the developed liposomal PTX formulations are troubled with low PTX encapsulation capacity (maximal content, 3 mol%) and accompanied by PTX crystallisation. The application of "pocket-forming" lipids significantly increased the encapsulation capacity of PTX in the liposomes up to 10 mol%. Stable lyophilised preparation of PTX (7 mol%) encapsulated in the liposomes composed of SOPC/POPG/MOPC (molar ratio, 60:20:20) doped with 5 mol% vitamin E had the size distribution of 180-190 nm (PDI, 0.1) with zeta-potential of -31 mV. Sucrose was found to be a suitable cryoprotectant at the lipid:sugar molar ratios of 1:5-1:10. This liposomal formulation did not show any evidence of toxicity in C57BL/6 mice treated with the highest doses of PTX (100 mg/kg administered as a single dose and 150 mg/kg as a cumulative dose applied in three equivalent doses in 48-h intervals). A dose dependent anticancer effect was found in both hollow fibre implants and syngenic B16F10 melanoma mouse tumour models. PMID- 19904828 TI - Pharmacokinetic/pharmacodynamic modeling and simulation to determine effective dosage regimens for doripenem. AB - The aim of this study was to obtain information on effective dosage regimens of doripenem by a modeling and simulation approach based on pharmacokinetic (PK)/pharmacodynamic (PD) theory. The PK/PD model we have already developed was modified to explain in vitro bactericidal kinetics of doripenem for several Pseudomonas aeruginosa strains. Time-course profiles of bacterial counts in patients infected with P. aeruginosa were simulated for typical clinical dosage regimens in Japan considering the variability of PK and the patients' backgrounds by a Monte Carlo simulation. Moreover, time-course profiles of probability achieving the criterion (log(CFU/mL) < 0) were predicted for the evaluation of antibacterial efficacy by renal function. The in vitro bacterial profiles at various dosage regimens could be well explained by the PK/PD model. The simulations suggested the dependence of antibacterial efficacy on the frequency of administration, indicating time-dependent antibacterial activity. It was also suggested that 500 mg t.i.d. showed significant bacterial reduction in patients for any degree of renal function and any severities in 2 weeks after the start of treatment. Our approach to simulate time-course profiles of bacterial counts should be useful for determining and examining effective dosage regimens, including the treatment period, in drug development. PMID- 19904830 TI - Excision of reprogramming transgenes improves the differentiation potential of iPS cells generated with a single excisable vector. AB - The residual presence of integrated transgenes following the derivation of induced pluripotent stem (iPS) cells is highly undesirable. Here we demonstrate efficient derivation of iPS cells free of exogenous reprogramming transgenes using an excisable polycistronic lentiviral vector. A novel version of this vector containing a reporter fluorochrome allows direct visualization of vector excision in living iPS cells in real time. We find that removal of the reprogramming vector markedly improves the developmental potential of iPS cells and significantly augments their capacity to undergo directed differentiation in vitro. We further propose that methods to efficiently excise reprogramming transgenes with minimal culture passaging, such as those demonstrated here, are critical since we find that iPS cells may acquire chromosomal abnormalities, such as trisomy of chromosome 8, similar to embryonic stem cells after expansion in culture. Our findings illustrate an efficient method for the generation of transgene-free iPS cells and emphasize the potential beneficial effects that may result from elimination of integrated reprogramming factors. In addition, our results underscore the consequences of long-term culture that will need to be taken into account for the clinical application of iPS cells. PMID- 19904829 TI - NOTCH pathway blockade depletes CD133-positive glioblastoma cells and inhibits growth of tumor neurospheres and xenografts. AB - Cancer stem cells (CSCs) are thought to be critical for the engraftment and long term growth of many tumors, including glioblastoma (GBM). The cells are at least partially spared by traditional chemotherapies and radiation therapies, and finding new treatments that can target CSCs may be critical for improving patient survival. It has been shown that the NOTCH signaling pathway regulates normal stem cells in the brain, and that GBMs contain stem-like cells with higher NOTCH activity. We therefore used low-passage and established GBM-derived neurosphere cultures to examine the overall requirement for NOTCH activity, and also examined the effects on tumor cells expressing stem cell markers. NOTCH blockade by gamma secretase inhibitors (GSIs) reduced neurosphere growth and clonogenicity in vitro, whereas expression of an active form of NOTCH2 increased tumor growth. The putative CSC markers CD133, NESTIN, BMI1, and OLIG2 were reduced following NOTCH blockade. When equal numbers of viable cells pretreated with either vehicle (dimethyl sulfoxide) or GSI were injected subcutaneously into nude mice, the former always formed tumors, whereas the latter did not. In vivo delivery of GSI by implantation of drug-impregnated polymer beads also effectively blocked tumor growth, and significantly prolonged survival, albeit in a relatively small cohort of animals. We found that NOTCH pathway inhibition appears to deplete stem-like cancer cells through reduced proliferation and increased apoptosis associated with decreased AKT and STAT3 phosphorylation. In summary, we demonstrate that NOTCH pathway blockade depletes stem-like cells in GBMs, suggesting that GSIs may be useful as chemotherapeutic reagents to target CSCs in malignant gliomas. PMID- 19904831 TI - The role of two putative nitroreductases, Frm2p and Hbn1p, in the oxidative stress response in Saccharomyces cerevisiae. AB - The nitroreductase family is comprised of a group of FMN- or FAD-dependent enzymes that are able to metabolize nitrosubstituted compounds using the reducing power of NAD(P)H. These nitroreductases can be found in bacterial species and, to a lesser extent, in eukaryotes. There is little information on the biochemical functions of nitroreductases. Some studies suggest their possible involvement in the oxidative stress response. In the yeast Saccharomyces cerevisiae, two nitroreductase proteins, Frm2p and Hbn1p, have been described. While Frm2p appears to act in the lipid signalling pathway, the function of Hbn1p is completely unknown. In order to elucidate the functions of Frm2p and Hbn1p, we evaluated the sensitivity of yeast strains, proficient and deficient in both oxidative stress proteins, for respiratory competence, antioxidant-enzyme activities, intracellular reactive oxygen species (ROS) production and lipid peroxidation. We found reduced basal activity of superoxide dismutase (SOD), ROS production, lipid peroxidation and petite induction and higher sensitivity to 4 nitroquinoline-oxide (4-NQO) and N-nitrosodiethylamine (NDEA), as well as higher basal activity of catalase (CAT) and glutathione peroxidase (GPx) and reduced glutathione (GSH) content in the single and double mutant strains frm2Delta and frm2Delta hbn1Delta. These strains exhibited less ROS accumulation and lipid peroxidation when exposed to peroxides, H(2)O(2) and t-BOOH. In summary, the Frm1p and Hbn1p nitroreductases influence the response to oxidative stress in S. cerevisae yeast by modulating the GSH contents and antioxidant enzymatic activities, such as SOD, CAT and GPx. PMID- 19904832 TI - Extraction of genotype-phenotype-drug relationships from text: from entity recognition to bioinformatics application. AB - Advances in concept recognition and natural language parsing have led to the development of various tools that enable the identification of biomedical entities and relationships between them in text. The aim of the Genotype Phenotype-Drug Relationship Extraction from Text workshop (or GPD-Rx workshop) is to examine the current state of art and discuss the next steps for making the extraction of relationships between biomedical entities integral to the curation and knowledge management workflow in Pharmacogenomics. The workshop will focus particularly on the extraction of Genotype-Phenotype, Genotype-Drug, and Phenotype-Drug relationships that are of interest to Pharmacogenomics. Extracting and structuring such text-mined relationships is a key to support the evaluation and the validation of multiple hypotheses that emerge from high throughput translational studies spanning multiple measurement modalities. In order to advance this agenda, it is essential that existing relationship extraction methods be compared to one another and that a community wide benchmark corpus emerges; against which future methods can be compared. The workshop aims to bring together researchers working on the automatic or semi-automatic extraction of relationships between biomedical entities from research literature in order to identify the key groups interested in creating such a benchmark. PMID- 19904833 TI - Infection, inflammation, and mental illness. The body's immune response to infectious agents may imperil the brain. PMID- 19904834 TI - Treating premenstrual dysphoric disorder. Options include drugs targeting serotonin and various types of hormone therapy. PMID- 19904835 TI - Lithium-induced kidney problems. Serious problems are unusual, but monitoring is key. PMID- 19904836 TI - Drug fails to subdue repetitive behavior in children with autism spectrum disorders. PMID- 19904838 TI - Health tips. Preventing bedsores. PMID- 19904837 TI - To prevent post-traumatic stress disorder, is it helpful to provide psychotherapy to everyone who has been exposed to a significant trauma? PMID- 19904839 TI - Traffic exposure--heart attack trigger? PMID- 19904840 TI - Prostatitis. The trouble with inflammation. PMID- 19904842 TI - What are the benefits of colon cleansing? I've seen a number of advertisements promoting it for good health and as a way to clear toxins from your body. PMID- 19904841 TI - Iron overload. Subtle signs, early treatment. PMID- 19904843 TI - [Pathology of salivary gland tumors]. PMID- 19904844 TI - [Total laryngectomy]. PMID- 19904845 TI - The new adult orphan: issues and considerations for health care professionals. AB - The death of the last parent has a profound effect on survivors. Health care workers are often the first source of anticipatory guidance for newly orphaned adults as they cope with grief, loss, and awareness that their lives are forever changed. It is estimated that more than 80 million Americans were born between 1946 and 1964. As this Baby Boomer generation, often defined as seeing themselves as culturally special, becomes "orphaned," they may be less aware, less prepared, and less supported than any previous group of Americans regarding this life event. For a number of adults, the loss creates many unexpected results that can destabilize life in profound ways. This article describes the unique new realities of helping adult orphans as they relate to health care providers and discusses the problems associated with prolonged and complicated grief. Implications for geriatric caregivers, mental health providers, health educators, and others are proposed. PMID- 19904846 TI - The 10 o'clock news. PMID- 19904850 TI - Health promotion course a valuable tool. PMID- 19904851 TI - Acute confusion/delirium. PMID- 19904852 TI - Palliative care for advanced dementia. AB - Advanced dementia is a terminal condition; however, research has revealed that affected individuals are subjected to multiple intrusive and burdensome interventions. Individuals with advance dementia require palliative care long before their death. This article reviews selected approaches that support personhood, explain behavior management principles, assure meaningful connections for activities of daily living, minimize weight loss, and explain advance directives and medical management best practices in palliative care for advanced dementia. PMID- 19904853 TI - Clearing the err. AB - Absent an infinitesimal percentage, most Americans seek health care services due to a legitimate health issue. Fundamental within this relationship is the understanding that health care professionals will do everything within their power and expertise to alleviate the suffering of each patient they treat. Unfortunately, preventable medical errors do occur, and the innocent patient is left to suffer. In 1999, the Institute of Medicine released To ERR Is Human: Building A Safer Health System, the first mainstream publication calling for a change in the culture of health care and the eradication of preventable medical errors. In the 10 years since its publication, federal and state governments and agencies have been proactive in attempting to meet the recommendations originally proposed in To ERR Is Human. This article will review what has been accomplished in this time frame. PMID- 19904854 TI - Reflection on photographs. AB - Nursing students' exposure to clinical placements with older adults is instrumental in helping them adopt positive attitudes toward care of that population. This qualitative pilot study analyzed perceptions and expectations of a group of first-year students prior to a clinical placement with older adults. A photo-elicitation technique, involving viewing of realistic photographs of older adults being cared for, was used to help students clarify expectations. This was followed by thematic analysis of their perceptions and expectations. Analysis revealed five main themes: Dissecting What It Means to Be a Nurse, Revisioning Therapeutic Relationships in Terms of Dignity, Youthful Reflection on the Differences Between Young and Old, Feeling Challenged and Confronted, and Experiencing Sensitivity and Awkwardness Toward Older Adults' Nakedness. Engagement with images of older adults encouraged students to anticipate their clinical placement in an aged care setting in a more meaningful, reflective way than they may have done without prior exposure, suggesting a need for realistic pre-practice education. PMID- 19904856 TI - Why not just ask the resident? AB - This research evaluated a draft preference assessment tool (draft-PAT) designed to replace the current Customary Routine section of the Minimum Data Set (MDS) for nursing homes. The draft-PAT was tested with a sample of nursing home residents to evaluate survey-level administration time and noncompletion rates, as well as item-level nonresponse rates, response distributions, and test-retest reliability. Modifications to the draft-PAT were then retested with a subsample of residents. Completion times were brief (generally less than 10 minutes), and only a small percentage of residents were unable to complete the interview. Item level nonresponse rates were low for the draft-PAT (0% to 8%) and even lower during retesting for items advanced to the national field trial (0% to 4%). Item response distributions indicated reasonable use of all options across both testing occasions, and item-level test-retest reliability was high. This study found that nursing home residents can reliably report their preferences. Eighteen items from the modified draft-PAT were advanced to the national field trial of the MDS 3.0. Inclusion of the PAT in the MDS revision underscores increased emphasis on including residents' voice in the assessment process. PMID- 19904858 TI - Our journey from a modular to a multimillion dollar education center: a rural hospital clinical education department's road to excellence. AB - This column discusses how a Midwest rural hospital's education department made the journey from a staff of four individuals located in a modular unit outside of the hospital to a multidisciplinary team reaching the hospital and the community from one of the most highly technologically advanced education centers in the nation. PMID- 19904859 TI - The EMPOWER project: an initiative to educate and mentor paraprofessional staff. AB - This column describes a new program designed to enhance patient safety by teaching paraprofessional staff (PPS) how to communicate changes in patient status to professional staff. Phase 1 of the project included assessment of PPS perceptions of the patient safety culture at the hospital through survey and focus group discussions. Results from Phase 1 and plans for Phase 2 are described. PMID- 19904860 TI - Teaching and learning in virtual worlds. AB - This column explores virtual worlds and their use in nursing. PMID- 19904861 TI - The effect of high-fidelity simulation training on medical-surgical graduate nurses' perceived ability to respond to patient clinical emergencies. AB - BACKGROUND: Recognition of and early intervention for patients with acutely deteriorating conditions is often the responsibility of medical-surgical nurses. This study examined the effect of simulation on medical-surgical graduate nurses' perceived ability and confidence in responding to patient clinical emergencies. METHOD: Fifty medical-surgical graduate students participated in high-fidelity immersive simulations. Questionnaires completed before and after simulation asked participants to rate their perceived ability and confidence. RESULTS: After simulation, participants reported increased confidence in their ability to perform both technical and nontechnical aspects of responding to patient clinical emergencies. Ninety-four percent of participants identified formal debriefing as the most useful aspect of the simulation experience. CONCLUSION: Medical-surgical graduate nurses' confidence and perceived technical and nontechnical skills during patient clinical emergencies are enhanced following simulation. The ability of graduates to transfer the increased confidence and perceived advanced resuscitation skills following simulation to the clinical environment needs to be investigated. PMID- 19904863 TI - Creating a culture of professional development: a milestone pathway tool for registered nurses. AB - The nursing shortage continues to be a significant threat to health care. Creating a culture of professional development in health care institutions is one way to combat this shortage. Professional development refers to a constant commitment to maintain one's knowledge and skill base. Increasing professional development opportunities in the health care setting has been shown to affect nurse retention and satisfaction. Several approaches have been developed to increase professional development among nurses. However, for the most part, these are "one size fits all" approaches that direct nurses to progress in lock step fashion in skill and knowledge acquisition within a specialty. This article introduces a milestone pathway tool for registered nurses designed to enhance professional development that is unique to the individual nurse and the specific nursing unit. This tool provides a unit-specific concept map, a milestone pathway template, and a personal professional development plan. PMID- 19904864 TI - Supporting preceptors: a three-pronged approach for success. AB - This article describes the development and implementation of the Nursing Preceptor Program (NPP), an innovative program that provides training and support to preceptors working with undergraduate nursing students. The main objective of the NPP is to support the learning outcomes of students by preparing and supporting preceptors for their roles. The University of Pittsburgh School of Nursing created a three-pronged program: a self-paced, modular, online preceptor course; a dedicated web page for both preceptors and students; and a relational database that can be queried for information on clinical sites and preceptors, catalogued by semester and year. A task force, consisting of local academic and clinical experts, was created to help with program development. To date, 25 preceptors have participated in the online course. Evaluations have been positive. Preceptors believe the included information is applicable and the modules, as a whole, are well planned and written. PMID- 19904865 TI - Fostering a smooth transition to the faculty role. AB - Both new and experienced nursing faculty across the United States are struggling to meet the individual needs of an increasingly diverse and growing population of nursing students, while simultaneously attempting to balance the research, scholarship, and stewardship requirements of their institutions. Faculty development is part of the professional career journey, and involves mentorship, guidance, and more formal educational experiences. One university's faculty development program is described and addresses the role of mentors and administrators in the faculty development process, the recognized need for continued development and partnering opportunities, and outcome data from two cohorts of new nursing faculty. PMID- 19904866 TI - Structured group reminiscence: an intervention for older adults. AB - Group reminiscence is an intervention recommended for care of older adults in structured and unstructured settings. One problem experienced by nurses is how to organize, facilitate, and evaluate reminiscence groups for older people. Hence, there is a need for further research on reminiscence to determine how to use this as an intervention to improve the well-being of older adults. There is also a need for continuing education to provide nurses with education on this intervention for older persons. This article provides an overview of qualitative and quantitative research on group reminiscence and offers a suggested evidence based protocol for a 6-week group intervention based on this research. PMID- 19904867 TI - The multigenerational household: implications for nursing education. PMID- 19904868 TI - Syllabus selection: innovative learning activity. PMID- 19904873 TI - Step-by-step practice transformation. PMID- 19904874 TI - Creating a high-quality, low-cost health care system: lessons from Grand Junction, Colorado. PMID- 19904875 TI - The 2009 EHR user satisfaction survey: responses from 2,012 family physicians. PMID- 19904876 TI - Ten steps to a patient-centered medical home. PMID- 19904877 TI - "Vital signs" for assessing your practice's financial health. PMID- 19904887 TI - What if Quiznos were run like health care? PMID- 19904889 TI - Is telmisartan effective for stroke prevention? PMID- 19904890 TI - Effects of marital status on cancer incidence and survival rates. PMID- 19904891 TI - Family physicians' present and future role in caring for older patients. AB - The population of patients older than 65 years is projected to increase substantially in the coming years, particularly in rural areas. Family physicians are essential providers of geriatric care, especially in rural areas, but need payment reform to improve their capacity to meet the needs of older patients. PMID- 19904892 TI - Endometrial cancer. AB - Endometrial cancer is the leading cause of gynecologic cancer in the United States. Etiologically, endometrial carcinoma usually results from unopposed estrogen stimulation of the endometrium, although non-estrogen-related forms occur as well. The most common presentation of endometrial cancer is postmenopausal bleeding. A variety of diagnostic modalities are available to aid in the detection of the disease, each with its own strengths and limitations. These modalities include endometrial biopsy, ultrasonography, saline infusion sonography, and hysteroscopy. A definitive diagnosis requires pathologic confirmation via endometrial biopsy or dilatation and curettage. Surgical staging of endometrial cancer will dictate how physicians manage the condition. For most women, staging and initial treatment are accomplished with total hysterectomy, bilateral salpingo-oophorectomy, and peritoneal washings. Surgery, radiation, and chemotherapy play a role in treatment, depending on tumor stage and grade. At present, there are no recommendations for screening the general population. PMID- 19904893 TI - Catheter ablation of supraventricular arrhythmias and atrial fibrillation. AB - Supraventricular arrhythmias are relatively common, often persistent, and rarely life-threatening cardiac rhythm disturbances that arise from the sinus node, atrial tissue, or junctional sites between the atria and ventricles. The term "supraventricular arrhythmia" most often is used to refer to supraventricular tachycardias and atrial flutter. The term "supraventricular tachycardia" commonly refers to atrial tachycardia, atrioventricular nodal reentrant tachycardia, and atrioventricular reciprocating tachycardia, an entity that includes Wolff Parkinson-White syndrome. Atrial fibrillation is a distinct entity classified separately. Depending on the arrhythmia, catheter ablation is a treatment option at initial diagnosis, when symptoms develop, or if medical therapy fails. Catheter ablation of supraventricular tachycardias, atrial flutter, and atrial fibrillation offers patients high effectiveness rates, durable (and often permanent) therapeutic end points, and low complication rates. Catheter ablation effectiveness rates exceed 88 percent for atrioventricular nodal reentrant tachycardia, atrioventricular reciprocating tachycardia, and atrial flutter; are greater than 86 percent for atrial tachycardia; and range from 60 to 80 percent for atrial fibrillation. Complication rates for supraventricular tachycardias and atrial flutter ablation are 0 to 8 percent. The complication rates for atrial fibrillation ablation range from 6 to 10 percent. Complications associated with catheter ablation result from radiation exposure, vascular access (e.g., hematomas, cardiac perforation with tamponade), catheter manipulation (e.g., cardiac perforation with tamponade, thromboembolic events), or ablation energy delivery (e.g., atrioventricular nodal block). PMID- 19904894 TI - Common forearm fractures in adults. AB - Fractures of the forearm are common injuries in adults. Proper initial assessment includes a detailed history of the mechanism of injury, a complete examination of the affected arm, and appropriate radiography. Open fractures, joint dislocation or instability, and evidence of neurovascular injury are indications for emergent referral. Fractures demonstrating significant displacement, comminution, or intra articular involvement may also warrant orthopedic consultation. In the absence of these findings, many forearm fractures can be managed by a primary care physician. Initial management of forearm fractures should follow the PRICE (protection, rest, ice, compression, and elevation) protocol, with the exception of compression, which should be avoided in the acute setting. Distal radius fractures with minimal displacement can be treated with a short arm cast. Isolated ulnar fractures can usually be managed with a short arm cast or a functional forearm brace. Mason type I radial head fractures can be treated with a splint for five to seven days or with a sling as needed for comfort, along with early range-of-motion exercises. Patients with an olecranon fracture are candidates for nonsurgical treatment if the elbow is stable and the extensor mechanism is intact. PMID- 19904895 TI - Tendinopathies of the foot and ankle. AB - Because our understanding of tendinopathy has evolved in recent years, the condition is now considered a degenerative process; this affects the approach to treatment. Initial therapy should always involve relative rest and modification of physical activity, use of rehabilitative exercises, and evaluation of intrinsic and extrinsic causes of injury. The posterior tibial tendon is a dynamic arch stabilizer; injury to this tendon can cause a painful flat-footed deformity with hindfoot valgus and midfoot abduction (characterized by the too many toes sign). Treatment of posterior tibial tendinopathy is determined by its severity and can include immobilization, orthotics, physical therapy, or subspecialty referral. Because peroneal tendinopathy is often misdiagnosed, it can lead to chronic lateral ankle pain and instability and should be suspected in a patient with either of these symptoms. Treatment involves physical therapy and close monitoring for surgical indications. Achilles tendinopathy is often caused by overtraining, use of inappropriate training surfaces, and poor flexibility. It is characterized by pain in the Achilles tendon 4 to 6 cm above the point of insertion into the calcaneus. Evidence from clinical trials shows that eccentric strengthening of the calf muscle can help patients with Achilles tendinopathy. Flexor hallucis longus tendinopathy is most common among ballet dancers. Patients may complain of an insidious onset of pain in the posteromedial aspect of the ankle; treatment involves correcting physical training errors, focusing on body mechanics, and strengthening the body's core. Anterior tibial tendinopathy is rare, but is typically seen in patients older than 45 years. It causes weakness in dorsiflexion of the ankle; treatment involves short-term immobilization and physical therapy. PMID- 19904897 TI - Nephrotic syndrome in adults: diagnosis and management. AB - Nephrotic syndrome may be caused by primary (idiopathic) renal disease or by a variety of secondary causes. Patients present with marked edema, proteinuria, hypoalbuminemia, and often hyperlipidemia. In adults, diabetes mellitus is the most common secondary cause, and focal segmental glomerulosclerosis and membranous nephropathy are the most common primary causes. Venous thromboembolism is a possible complication; acute renal failure and serious bacterial infection are also possible, but much less common. There are no established guidelines on the diagnostic workup or management of nephrotic syndrome. Imaging studies are generally not needed, and blood tests should be used selectively to diagnose specific disorders rather than for a broad or unguided workup. Renal biopsy may be useful in some cases to confirm an underlying disease or to identify idiopathic disease that is more likely to respond to corticosteroids. Treatment of most patients should include fluid and sodium restriction, oral or intravenous diuretics, and angiotensin-converting enzyme inhibitors. Some adults with nephrotic syndrome may benefit from corticosteroid treatment, although research data are limited. Intravenous albumin, prophylactic antibiotics, and prophylactic anticoagulation are not currently recommended. PMID- 19904896 TI - Saline nasal irrigation for upper respiratory conditions. AB - Saline nasal irrigation is an adjunctive therapy for upper respiratory conditions that bathes the nasal cavity with spray or liquid saline. Nasal irrigation with liquid saline is used to manage symptoms associated with chronic rhinosinusitis. Less conclusive evidence supports the use of spray and liquid saline nasal irrigation to manage symptoms of mild to moderate allergic rhinitis and acute upper respiratory tract infections. Consensus guidelines recommend saline nasal irrigation as a treatment for a variety of other conditions, including rhinitis of pregnancy and acute rhinosinusitis. Saline nasal irrigation appears safe, with no reported serious adverse events. Minor adverse effects can be avoided with technique modification and salinity adjustment. PMID- 19904899 TI - Screening for type 2 diabetes mellitus in adults. PMID- 19904900 TI - Headache in a well-appearing young woman. Idiopathic intracranial hypertension. PMID- 19904901 TI - Diagnosing lumbar spinal stenosis. PMID- 19904904 TI - Reactivity of halo(pyridinium)carbenes. AB - The reactivity of chloro- and fluoro(N-methyl-3-pyridinium)carbenes was examined by laser flash photolysis, where the halo(pyridinium)carbenes formed ylides with pyridine, acetonitrile, and acetone. Although the halo(pyridinium)carbenes reacted within the time of the laser pulse, their relative reactivities with a series of alkenes could be obtained from quenching experiments by using carbene pyridine ylides. Their relative order of reactivity with the alkenes and their poor overall selectivity showed that the halo(pyridinium)carbenes are strongly reactive electrophilic species. Computational studies demonstrated that the alkene (HOMO)-carbene (LUMO) interaction is predominant in halo(pyridinium)carbene-alkene reactions, supporting the electrophilic nature of these species. PMID- 19904906 TI - Liquid-crystalline nematic phase in aqueous suspensions of a disk-shaped natural beidellite clay. AB - After size-selection and osmotic pressure measurements at fixed ionic strength, the behavior of aqueous colloidal suspensions of anisotropic disklike beidellite clay particles has been investigated by combining optical observations under polarized light, rheological, and small angle X-ray scattering (SAXS) experiments. The obtained phase diagrams (volume fraction/ionic strength) reveal, for ionic strength below 10(-3) M/L, a first-order isotropic/nematic (I/N) phase transition before gel formation at low volume fractions, typically around 0.5%. This I/N transition line displays a positive slope for increasing ionic strength and shifts toward lower volume fraction with increasing particle size, confirming that the system is controlled by repulsive interactions. The swelling laws, derived from the interparticle distances obtained by SAXS, display a transition from isotropic swelling at low volume fractions to lamellar swelling at higher volume fractions. The liquid-crystal properties have then been investigated in detail. Highly aligned nematic samples can be obtained in three different ways, by applying a magnetic field, an ac electric field, and by spontaneous homeotropic anchoring on surfaces. The birefringence of the fluid nematic phase is negative with typical values around 5 x 10(-4) at a volume fraction of about 0.6%. High nematic order parameters have been obtained as expected for well aligned samples. The nematic director is aligned parallel to the magnetic field and perpendicular to the electric field. PMID- 19904905 TI - Lewis acid catalyzed indole synthesis via intramolecular nucleophilic attack of phenyldiazoacetates to iminium ions. AB - Lewis acids catalyze the cyclization of methyl phenyldiazoacetates with an ortho imino group, prepared from o-aminophenylacetic acid, to give 2,3-substituted indoles in quantitative yields. PMID- 19904907 TI - A spectroscopic and computational investigation of the conformational structural changes induced by hydrogen bonding networks in the glycidol-water complex. AB - Rotational spectra were recorded in natural abundance for the (13)C isotopomers of two conformers of glycidol. Moments of inertia from the (13)C isotopomers were used to calculate the substitution coordinates and C-C bond lengths of two glycidol monomer conformations. The structures of seven different conformational minima were found from ab initio (MP2/6-311++G(d,p)) optimizations of glycidol water. The rotational spectrum of glycidol-water was recorded using microwave spectroscopy, and the rotational constants were determined to be A = 3902.331 (11) MHz, B = 2763.176 (3) MHz, and C = 1966.863 (3) MHz. Rotational spectra were also recorded for glycidol-H(2)(18)O, glycidol-D(b)OH, and glycidol-d(O)-D(2)O. The rotational spectra were assigned to the lowest-energy ab initio structure, and the structure was improved by fitting to the experimental moments of inertia. The best-fit structure shows evidence for structural changes in glycidol to accommodate formation of the intermolecular hydrogen bonding network: the O-C-C-O torsional angle in glycidol was found to increase from 40.8 degrees for the monomer to 49.9 degrees in the water complex. PMID- 19904908 TI - Molecular tuning of phenylene-vinylene derivatives for two-photon photosensitized singlet oxygen production. AB - Substituent-dependent features and properties of the sensitizer play an important role in the photosensitized production of singlet oxygen, O(2)(a(1)Delta(g)). In this work, we systematically examine the effect of molecular changes in the sensitizer on the efficiency of singlet oxygen production using, as the sensitizer, oligophenylene-vinylene derivatives designed to optimally absorb light in a nonlinear two-photon process. We demonstrate that one cannot always rely on rule-of-thumb guidelines when attempting to construct efficient two photon singlet oxygen sensitizers. Rather, as a consequence of behavior that can deviate from the norm, a full investigation of the photophysical properties of the system is generally required. For example, it is acknowledged that the introduction of a ketone moiety to the sensitizer chromophore often results in more efficient production of singlet oxygen. However, we show here that the introduction of a carbonyl into a given phenylene-vinylene can, rather, have adverse effects on the yield of singlet oxygen produced. Using these molecules, we show that care must also be exercised when using qualitative symmetry-derived arguments to predict the relationship between one-and two-photon absorption spectra. PMID- 19904909 TI - Enantioselective total synthesis of (-)-acutumine. AB - An account of the total synthesis of the tetracyclic alkaloid (-)-acutumine is presented. A first-generation approach to the spirocyclic subunit was unsuccessful as a result of incorrect regioselectivity in a radical cyclization. However, this work spawned a second-generation strategy in which the spirocycle was fashioned via a radical-polar crossover reaction. This process merged an intramolecular radical conjugate addition with an enolate hydroxylation and created two stereocenters with excellent diastereoselectivity. The reaction was promoted by irradiation with a sunlamp, and a ditin reagent was required for aryl radical formation. These facts suggest that the substrate may function as a sensitizer, thereby facilitating homolytic cleavage of the ditin reagent. The propellane motif of the target was then installed via annulation of a pyrrolidine ring onto the spirocycle. The sequence of reactions used included a phenolic oxidation, an asymmetric ketone allylation mediated by Nakamura's chiral allylzinc reagent, an anionic oxy-Cope rearrangement, a one-pot ozonolysis reductive amination, and a Lewis acid promoted cyclization of an amine onto an alpha,beta-unsaturated dimethyl ketal. Further studies of the asymmetric ketone allylation demonstrated the ability of the Nakamura reagent to function well in a mismatched situation. A TiCl(4)-catalyzed regioselective methyl enol etherification of a 1,3-diketone completed the synthesis. PMID- 19904911 TI - "Self-preservation" of CO(2) gas hydrates--surface microstructure and ice perfection. AB - Gas hydrates can exhibit an anomalously slow decomposition outside their thermodynamic stability field; the phenomenon is called "self-preservation" and is mostly studied at ambient pressure and at temperatures between approximately 240 K and the melting point of ice. Here, we present a combination of in situ neutron diffraction studies, pVT work, and ex situ scanning electron microscopy (SEM) on CO(2) clathrates covering a much broader p-T field, stretching from 200 to 270 K and pressures between the hydrate stability limit and 0.6 kPa (6 mbar), a pressure far outside stability. The self-preservation regime above 240 K is confirmed over a broad pressure range and appears to be caused by the annealing of an ice cover formed in the initial hydrate decomposition. Another, previously unknown regime of the self-preservation exists below this temperature, extending however only over a rather narrow pressure range. In this case, the initial ice microstructure is dominated by a fast two-dimensional growth covering rapidly the clathrate surface. All observations lend strong support to the idea that the phenomenon of self-preservation is linked to the permeability of the ice cover governed by (1) the initial microstructure of ice and/or (2) the subsequent annealing of this ice coating. The interplay of the microstructure of newly formed ice and its annealing with the ongoing decomposition reaction leads to various decomposition paths and under certain conditions to a very pronounced preservation anomaly. PMID- 19904910 TI - Noncovalent interactions involving histidine: the effect of charge on pi-pi stacking and T-shaped interactions with the DNA nucleobases. AB - Detailed (gas-phase) MP2/6-31G*(0.25) potential energy surface scans and CCSD(T) energy calculations at the complete basis set (CBS) limit were used to analyze the (face-to-face) stacking and (edge-to-face) T-shaped interactions between histidine (modeled as imidazole) and the DNA nucleobases. For the first time, a variety of relative monomer arrangements between both neutral and protonated histidine and the natural nucleobases were considered to determine the effects of charge on the optimum dimer geometry and binding strength. Our results reveal that protonation of histidine changes the preferred relative orientations of the monomers and propose that these geometric differences may be combined with experimental crystal structures to assess the protonation state of histidine in different environments. It is also found that protonation affects the nucleobase binding preference, as well as the magnitude of the stacking and T-shaped interactions. Indeed, the maximum possible stacking and T-shaped interactions involving the neutral histidine range between approximately 20 and 45 kJ mol(-1), while this range increases to 40-105 kJ mol(-1) upon protonation, which represents an up to 330% enhancement. Although an increase in the interaction energies upon protonation of histidine is expected, the present work provides a measure of the magnitude of this enhancement in the gas phase and reveals that the amplification is almost entirely due to larger electrostatic contributions. The relative strengthening of different classifications of dimers upon protonation leads to stronger T-shaped interactions than stacking energies for protonated histidine, while the stacking and T-shaped interactions involving neutral histidine are of comparable magnitude. Thus, there is a significant difference in the nature of the pi(cation)-pi interactions involving protonated histidine and the pi-pi interactions involving neutral histidine. The calculated strengths of the interactions studied in the present work suggest that both neutral and cationic histidine contacts will provide significant stabilization to DNA-protein complexes. Although solvation effects will decrease the magnitude of the reported interactions, our results are applicable to a variety of low polarity, biologically-relevant environments such as nonpolar enzyme active sites. Therefore, our calculations suggest that these interactions may also be important for many biological processes. The proposed significance of these interactions is supported by the large number of histidine-nucleobase contacts that appear in experimental crystal structures. The highly accurate (MP2/6 31G*(0.25)) preferred structures and (CCSD(T)/CBS) binding strengths reported in the present work can be used as benchmarks to analyze the performance of existing, or to develop new, molecular mechanics force fields for use in large scale molecular dynamics (MD) studies of DNA-protein complexes. PMID- 19904912 TI - Total mass difference statistics algorithm: a new approach to identification of high-mass building blocks in electrospray ionization Fourier transform ion cyclotron mass spectrometry data of natural organic matter. AB - The ultrahigh-resolution Fourier transform ion cyclotron resonance (FTICR) mass spectrum of natural organic matter (NOM) contains several thousand peaks with dozens of molecules matching the same nominal mass. Such a complexity poses a significant challenge for automatic data interpretation, in which the most difficult task is molecular formula assignment, especially in the case of heavy and/or multielement ions. In this study, a new universal algorithm for automatic treatment of FTICR mass spectra of NOM and humic substances based on total mass difference statistics (TMDS) has been developed and implemented. The algorithm enables a blind search for unknown building blocks (instead of a priori known ones) by revealing repetitive patterns present in spectra. In this respect, it differs from all previously developed approaches. This algorithm was implemented in designing FIRAN-software for fully automated analysis of mass data with high peak density. The specific feature of FIRAN is its ability to assign formulas to heavy and/or multielement molecules using "virtual elements" approach. To verify the approach, it was used for processing mass spectra of sodium polystyrene sulfonate (PSS, M(w) = 2200 Da) and polymethacrylate (PMA, M(w) = 3290 Da) which produce heavy multielement and multiply-charged ions. Application of TMDS identified unambiguously monomers present in the polymers consistent with their structure: C(8)H(7)SO(3)Na for PSS and C(4)H(6)O(2) for PMA. It also allowed unambiguous formula assignment to all multiply-charged peaks including the heaviest peak in PMA spectrum at mass 4025.6625 with charge state 6- (mass bias 0.33 ppm). Application of the TMDS-algorithm to processing data on the Suwannee River FA has proven its unique capacities in analysis of spectra with high peak density: it has not only identified the known small building blocks in the structure of FA such as CH(2), H(2), C(2)H(2)O, O but the heavier unit at 154.027 amu. The latter was identified for the first time and assigned a formula C(7)H(6)O(4) consistent with the structure of dihydroxyl-benzoic acids. The presence of these compounds in the structure of FA has so far been numerically suggested but never proven directly. It was concluded that application of the TMDS-algorithm opens new horizons in unfolding molecular complexity of NOM and other natural products. PMID- 19904913 TI - Naphthalimide-porphyrin hybrid based ratiometric bioimaging probe for Hg2+: well resolved emission spectra and unique specificity. AB - In this paper, we unveil a novel naphthalimide-porphyrin hybrid based fluorescence probe (1) for ratiometric detection of Hg(2+) in aqueous solution and living cells. The ratiometric signal change of the probe is based on a carefully predesigned molecule containing two independent Hg(2+)-sensitive fluorophores with their maximal excitation wavelengths located at the same range, which shows reversibly specific ratiometric fluorescence responses induced by Hg(2+). In the new developed sensing system, the emissions of the two fluorophores are well-resolved with a 125 nm difference between two emission maxima, which can avoid the emission spectra overlap problem generally met by spectra-shift type probes and is especially favorable for ratiometric imaging intracellular Hg(2+). It also benefits from a large range of emission ratios and thereby a high sensitivity for Hg(2+) detection. Under optimized experimental conditions, the probe exhibits a stable response for Hg(2+) over a concentration range from 1.0 x 10(-7) to 5.0 x 10(-5) M, with a detection limit of 2.0 x 10(-8) M. The response of the probe toward Hg(2+) is reversible and fast (response time less than 2 min). Most importantly, the ratiometric fluorescence changes of the probe are remarkably specific for Hg(2+) in the presence of other abundant cellular metal ions (i.e., Na(+), K(+), Mg(2+), and Ca(2+)), essential transition metal ions in cells (such as Zn(2+), Fe(3+), Fe(2+), Cu(2+), Mn(2+), Co(2+), and Ni(2+)), and environmentally relevant heavy metal ions (Ag(+), Pb(2+), Cr(3+), and Cd(2+)), which meets the selective requirements for biomedical and environmental monitoring application. The recovery test of Hg(2+) in real water samples demonstrates the feasibility of the designed sensing system for Hg(2+) assay in practical samples. It has also been used for ratiometric imaging of Hg(2+) in living cells with satisfying resolution, which indicates that our novel designed probe has effectively avoided the general emission spectra overlap problem of other ratiometric probes. PMID- 19904914 TI - Scanning mass spectrometry probe: a scanning probe electrospray ion source for imaging mass spectrometry of submerged interfaces and transient events in solution. AB - The scanning mass spectrometry (SMS) probe is a new electrospray ion source. Motivated by the need for untargeted chemical imaging of dynamic events in solution, we have exploited an approach to electrospray ionization (ESI) that allows continuous sampling from a highly localized volume (approximately picoliters) in a liquid environment, softly ionizes molecules in the sample to render them amenable for mass spectrometric analysis, and sends the ions to the mass spectrometer. The key underlying concepts for our approach are (1) treating the electrospray capillary inlet as a chemical scanning probe and (2) locating the electrospray point as close as possible to the sampling point, thus providing the shortest response time possible. This approach enables chemical monitoring or imaging of submerged interfaces, providing access to details of spatial heterogeneity and temporal changes within liquid samples. It also permits direct access to liquid/ liquid interfaces for ESI-MS analysis. In this letter we report the first demonstrations of these capabilities of the SMS probe and describe some of the probe's basic characteristics. PMID- 19904915 TI - Matrix-assisted laser desorption/ionization mass spectrometry method for selectively producing either singly or multiply charged molecular ions. AB - Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is noted for its ability to produce primarily singly charged ions. This is an attribute when using direct ionization for complex mixtures such as protein digests or synthetic polymers. However, the ability to produce multiply charged ions, as with electrospray ionization (ESI), has advantages such as extending the mass range on mass spectrometers with limited mass-to-charge (m/z) range and enhancing fragmentation for structural characterization. We designed and fabricated a novel field free transmission geometry atmopsheric pressure (AP) MALDI source mounted to a high-mass resolution Orbitrap Exactive mass spectrometer. We report the ability to produce at will either singly charged ions or highly charged ions using a MALDI process by simply changing the matrix or the matrix preparation conditions. Mass spectra with multiply charged ions very similar to those obtained with ESI of proteins such as cytochrome c and ubiquitin are obtained with low femtomole amounts applied to the MALDI target plate and for peptides such as angiotensin I and II with application of attomole amounts. Single scan acquisitions produce sufficient ion current even from proteins. PMID- 19904917 TI - Three-dimensional frameworks of cubic (NH4)5Ga4SbS10, (NH4)4Ga4SbS9(OH) x H2O, and (NH4)3Ga4SbS9(OH2) x 2 H2O. AB - Three new isostructural open-framework sulfides, (NH(4))(5)Ga(4)SbS(10) (1), (NH(4))(4)Ga(4)SbS(9)(OH) x H(2)O (2a), and (NH(4))(3)Ga(4)SbS(9)(H(2)O) x 2 H(2)O (2b), were synthesized under basic hydrothermal conditions using ammonium hydroxide as the structure-directing agent. The structures feature a three dimensional open framework comprised of adamantane [Ga(4)Q(10)](8-) clusters linked with Sb(3+) centers. The compounds are wide gap semiconductors, crystallize in the chiral space group P2(1)3, and represent a new structure type. They exhibit nonlinear optical properties. PMID- 19904916 TI - Site-preferential dissociation of peptides with active chemical modification for improving fragment ion detection. AB - Multiple reaction monitoring tandem mass spectrometry becomes an important strategy for measuring protein targets in complex biomatrixes. Active chemical modification of peptides like phenylthiocarbamoylation has unique potential for improving the measurement. This potential is enabled by active participation of a modifying group in site-preferential dissociation of modified peptides, which produces certain fragment ions at very high yields and in a sequence-independent manner. In this work, a novel combination of energy-resolved mass spectrometry with substituent effect investigation is used to analyze important factors that control the specificity of the site-preferential dissociation of phenylthiocarbamoyl peptides. On the basis of the linear correlation between collision energy and the Hammett constant as well as computational studies, it is found that the initial enhanced capture of a mobile proton and the subsequent, site-directed intramolecular proton transfer are important to the high yields (approximately 70-90%) for producing two types of fragment ions of phenylthiocarbamoyl peptides: the modified b(1) ion and the complementary y(n-1) ion. This understanding will help the design of new modification reagents. When integrated with the throughput and the signal-enhancing potential of peptide modification, active chemical modification of peptides will significantly advance mass spectrometry-based, targeted proteome analysis. PMID- 19904918 TI - Fabrication and electrical properties of si nanowires synthesized by Al catalyzed vapor-liquid-solid growth. AB - The synthesis of epitaxially oriented Si nanowires at high growth rates (>1 microm/min) was demonstrated on (111) Si substrates using Al as the catalyst. The use of high H(2) and SiH(4) partial pressures was found to be effective at reducing problems associated with Al oxidation and nanowire nucleation, enabling growth of high aspect ratio structures at temperatures ranging from 500 to 600 degrees C with minimal tapering of the diameter. Because of the high growth rate observed, the Al catalyst is believed to be in the liquid state during the growth. Four-point resistance measurements and back-gated current-voltage measurements indicate that the wires are p-type with an average resistivity of 0.01 +/- 0.004 Omega-cm. These results suggest that Al is incorporated into the Si nanowires under these conditions at concentrations higher than the solubility limit (5-6 x 10(18) cm(-3)) for Al in Si at 550 degrees C. This work demonstrates that Al can serve as both an effective catalyst and p-type dopant for the growth of Si nanowires. PMID- 19904919 TI - Bioactive constituents of the stem bark of Beilschmiedia zenkeri. AB - Phytochemical investigation of the stem bark of Beilschmiedia zenkeri led to the isolation of four new methoxylated flavonoid derivatives, (2S,4R)-5,6,7 trimethoxyflavan-4-ol (1), (2S,4R)-4,5,6,7-tetramethoxyflavan (2), beilschmieflavonoid A (3), and beilschmieflavonoid B (4), together with seven known compounds. The structures of 1-4 were established by spectroscopic methods, and their relative configurations confirmed by X-ray crystallographic and CD analysis. The isolated compounds were evaluated in vitro for their antibacterial activity against three strains of bacteria, Pseudomonas agarici, Bacillus subtilis, and Streptococcus minor, and for their antiplasmodial activity against Plasmodium falciparum, chloroquine-resistant strain W2. PMID- 19904920 TI - Spatial resolution of a type II heterojunction in a single bipolar molecule. AB - Bipolar molecules incorporating donor and acceptor components within a single molecule create exciting device opportunities due to their possible use as nanoscale p-n heterojunctions. Here we report a direct characterization of the internal electronic structure of a single bipolar molecular heterojunction, including subnanometer features of the intramolecular donor-acceptor interface. Angstrom-resolved scanning tunneling spectroscopy was used to map the energy levels and spatial extent of molecular orbitals across the surface of an individual bipolar molecule, bithiophene naphthalene diimide (BND). We find that individual BND molecules exhibit type II heterojunction behavior with orbital energy shifts occurring over subnanometer intramolecular interface distances. Comparison of this behavior with first-principles theoretical modeling provides new insights into the optimization of these molecular systems. PMID- 19904922 TI - A new intermolecular interaction: unconventional hydrogen bonds with element hydride bonds as proton acceptor. PMID- 19904921 TI - Benzo[cd]azulene skeleton: azulene, heptafulvene, and tropone derivatives. AB - Due to our interest in protein kinase modulating compounds, we developed syntheses for benzo[cd]azulenes. By using very common catalysts and reagents, such as t-BuOK, HCl, and mCPBA, the commercially available guaiazulene is converted in three steps into tricyclic tropone derivatives. Electrophilic aromatic substitution reactions of guaiazulene proceed without a catalyst. Complex one-pot reactions convert 1'-hydroxyalkyl azulenes into tricyclic heptafulvenes, and finally, the mild oxidant mCPBA cleaves the semicyclic C horizontal lineC double bonds to furnish tropones. PMID- 19904923 TI - Selective synthesis of eight-membered cyclic ureas by the [6 + 2] cycloaddition reaction of 2-vinylazetidines and electron-deficient isocyanates. AB - The [6 + 2] cycloaddition reaction of 2-vinylazetidines with electron-deficient isocyanates such as tosyl isocyanate proceeded smoothly in the absence of the catalyst at room temperature, and various cyclic ureas were isolated in good to high yields. Electron-deficient allenes also reacted with the 2-vinylazetidine, and the corresponding azocine derivatives were isolated. PMID- 19904924 TI - 4-Fluoro-2,4-methanoproline. AB - The first fluorinated analogue of the naturally occurring 2,4-methanoproline, 4 fluoro-2,4-methanoproline, has been synthesized in five steps from commercially available methyl 2-fluoroacrylate through a photochemical cyclization as a key step in generating a 2-azabicyclo[2.1.1]hexane skeleton. PMID- 19904925 TI - An unexpected highly stereoselective bisaziridination of (E,E)-1,4-dialkyl-2,3 dinitrobutadienes followed by a nitro group driven ring enlargement. AB - (+/-)-2,2'-Dinitro-2,2'-biaziridines were obtained by a direct aza-MIRC (Michael initiated ring closure) reaction on (E,E)-1,4-dialkyl-2,3-dinitro-l,3-butadienes under very mild conditions. The reactions occur with high stereoselectivity as shown by the enantioselective HPLC analyses performed on the crude mixtures. Ring enlargement to 3,3'-bi(1,2,4-oxadiazole) derivatives was easily obtained by a simple treatment with sodium iodide in DMSO, with an unforeseen regioselective aziridine C-C cleavage. PMID- 19904926 TI - Boron-catalyzed direct aldol reactions of pyruvic acids. AB - Interactions between pyruvic acids and diphenylborinic acid form the basis of an efficient, direct, boron-catalyzed aldol reaction that takes place in water at room temperature with low catalyst loadings. Both boronic and borinic acids function as catalysts, with the latter demonstrating particularly high activity. A wide range of aldehydes, including enolizable species, may be employed, delivering useful isotetronic acid derivatives in high yields. PMID- 19904927 TI - Synthesis of benzo[c] and naphtho[c]heterocycle diesters and dinitriles via homoelongation. AB - Syntheses of benzo[c] and naphtho[c]heterocycle diesters and dinitriles were achieved via our newly developed iterative elongation protocol. The photophysical and electrochemical properties of these conjugated systems are explored. PMID- 19904928 TI - Structural relaxation around Cr3+ in YAlO3-YCrO3 perovskites from electron absorption spectra. AB - The structural relaxation around Cr(3+) in YAl(1-x)Cr(x)O(3) perovskites was investigated and compared with analogous Cr-Al joins (corundum, spinel, garnet). Eight compositions (x(Cr)((3+)) from 0 to 1) were prepared by sol-gel combustion and were analyzed by a combined X-ray diffraction (XRD) and electron absorption spectroscopy (EAS) approach. The unit cell parameters and the XRD averaged octahedral (Cr,Al)-O and ([VIII])Y-O bond distances scale linearly with the chromium fraction. The optical parameters show an expected decrease of crystal field strength (10Dq) and an increase of covalency (B(35)) and polarizability (B(55)) toward YCrO(3), but a nonlinear trend outlines some excess 10Dq below x(Cr)((3+)) approximately 0.4. The local Cr-O bond lengths, as calculated from EAS, indicate a compression from 1.98 A (x(Cr)((3+)) = 1.0) down to 1.95 A (x(Cr)((3+)) = 0.035) so that the relaxation coefficient of perovskite (epsilon = 0.54) is the lowest in comparison with garnet (epsilon = 0.74), spinel (epsilon = 0.68), and corundum (epsilon = 0.58) in contrast with its structural features. The enhanced covalent character of the Cr(3+)-O-Cr(3+) bond in the one dimensional arrangement of corner-sharing octahedra can be invoked as a factor limiting the perovskite polyhedral network flexibility. The increased probability of Cr-O-Cr clusters for x(Cr)((3+)) greater than approximately 0.4 is associated to diverging trends of nonequivalent interoctahedral angles. The relatively low relaxation degree of Y(Al,Cr)O(3) can be also understood by considering an additional contribution to 10Dq because of the electrostatic potential of the rest of the lattice ions upon the localized electrons of the CrO(6) octahedron. Such an "excess" of 10Dq increases when the point symmetry of the Cr site is low, as in perovskite, and would be affected by the change of yttrium effective coordination number observed by XRD for x(Cr)((3+)) greater than approximately 0.4. This would justify the systematic underestimation of local Cr-O bond distances, as inferred from EAS, compared to what is derived from X-ray absorption (XAS) studies implying a stronger degree of relaxation around Cr(3+) of all the structures considered and supporting the hypothesis that 10Dq from EAS contains more information than previously retained particularly an additional contribution from the next nearest neighboring ions. PMID- 19904929 TI - Discovery of (Z)-5-(4-methoxybenzylidene)thiazolidine-2,4-dione, a readily available and orally active glitazone for the treatment of concanavalin A-induced acute liver injury of BALB/c mice. AB - A large amount of evidence suggests that monocytes/macrophages infiltration is implicated in a variety of inflammatory diseases including acute liver injury. Monocyte chemoattractant protein 1 (MCP-1) plays a crucial role in the process of macrophages recruitment. We herein presented a small-molecule library and a feasible quick screening method of evaluating potency of inhibition of chemotaxis of RAW264.7 cells stimulated by MCP-1. Fifty-three small molecules were synthesized and screened, and four compounds (2g, 2h, 4f, and 6h) showed inhibitory effects with IC(50) values range from 0.72 to 20.47 microM, with compound 4f being the most efficient. Further in vivo studies demonstrated that oral administration of 2g, 2h, 4f, or 6h decreases, most significantly for 4f, the serum levels of alanine aminotransaminase (ALT) and asparate aminotransaminase (AST) in ConA-induced acute livery injury BALB/c mice. Histopathological evaluation liver sections confirmed 4f as a potent, orally active compound for hepatoprotective effects against ConA-induced acute liver injury in BALB/c mice. PMID- 19904930 TI - 1H NMR-based metabonomics for the classification of Greek wines according to variety, region, and vintage. Comparison with HPLC data. AB - A sensitive and simple method was developed for the classification of wines according to variety, geographical origin, and vintage using NMR-based metabonomics. Polyphenol-rich extracts were prepared from 67 varietal wines from the principal wine-producing regions of Greece, using adsorption resin XAD-4. 1D (1)H NMR spectra obtained from the corresponding extracts were segmented, integrated, and normalized, and the data were subjected to principal component analysis. The chemometric classification of wines according to their phenolic profile allows discrimination between wines from different wineries of the same wine-producing zone and between different vintages for wines of the same variety. PMID- 19904932 TI - Determination of pesticide residues in integrated pest management and nonintegrated pest management samples of apple (Malus pumila Mill.). AB - Studies were undertaken to analyze the residues of commonly used pesticides viz. chlorpyrifos, endosulfan, dicofol, cypermethrin, fenvalerate, propargite, malathion, phorate, carbendazim, carbosulfan, thiamethoxam, and mancozeb in apple of integrated pest management (IPM) and non-IPM samples collected from the IPM and non-IPM fields of Shimla. We also present a method for the determination of these pesticides in apple samples. Residues of chlorpyrifos, endosulfan, dicofol, cypermethrin, fenvalerate, and propargite were analyzed by gas chromatography, while residues of carbendazim, carbosulfan, and thiamethoxam were analyzed by high-performance liquid chromatography. Residues of mancozeb were determined by a colorimetric method. Recoveries of all of the pesticides ranged from 61.30 to 95.46% at 0.1, 0.2, and 1.0 microg g(-1) levels of fortification with relative standard deviations ranging between 0.8 and 8.7. Apples from IPM and non-IPM orchards were analyzed for these pesticides using a developed method. Except for carbendazim and chlorpyrifos, the residues of all of the pesticides analyzed were below detectable limits. Although residues of carbendazim and chlorpyrifos were below the prescribed limits of maximum residue levels in both IPM and non-IPM orchards, residues were lower in apples from IPM orchards. PMID- 19904933 TI - Relationship between dieldrin uptake in cucumber and solvent-extractable residue in soil. AB - To prevent the distribution of cucumbers with dieldrin contamination exceeding the limit set by the Japanese Food Sanitation Law, the extraction solvent for dieldrin-contaminated soil was selected prior to cultivation so that the dieldrin residue level in cucumber could be predicted. The exhaustive extraction from soil could not explain the dieldrin uptake by cucumber plants. However, significant correlation (R(2) = 0.966, P < 0.001) was observed between dieldrin concentrations in cucumber and dieldrin concentrations extracted with 50% (v/v) methanol-water solution from soils. This was a result of the phytoavailability of dieldrin to the cucumber plants. The extractability of soil dieldrin with the methanol-water solution decreased as the organic carbon content in the soils increased. This suggested that a 50% (v/v) methanol-water solution is the optimal solution for predicting dieldrin concentrations in cucumbers by soil analysis. PMID- 19904934 TI - 4-Oxo-beta-lactams (azetidine-2,4-diones) are potent and selective inhibitors of human leukocyte elastase. AB - Human leukocyte elastase (HLE) is a serine protease stored in and secreted from neutrophils that plays a determinant role in the pathogenesis of several lung diseases. 4-Oxo-beta-lactams, previously reported as acylating agents of porcine pancreatic elastase, were found to be selective and potent inhibitors of HLE. Structure-activity relationship analysis showed that inhibitory activity is very sensitive to the nature of C-3 substituents, with small alkyl substituents such as a gem-diethyl group improving the inhibitory potency when compared to gem methyl benzyl or ethyl benzyl counterparts. 4-Oxo-beta-lactams containing a heteroarylthiomethyl group on the para position of an N(1)-aryl moiety afforded highly potent and selective inhibition of HLE, even at a very low inhibitor to enzyme ratio, as shown by the k(on) value of 3.24 x 10(6) M(-1) s(-1) for 6f. The corresponding ortho isomers were 40- to 90-fold less potent. PMID- 19904935 TI - Antioxidant and alpha-glucosidase inhibitory activity of colored grains in China. AB - Colored grains including red, purple, and black rice, purple corn, black barley, and black soybean contain anthocyanins. The present study was designed to (i) identify and quantify the individual anthocyanins and measure the total phenolic content (TPC), (ii) evaluate the antioxidant and alpha-glucosidase inhibitory activity, and (iii) correlate the TPC with total antioxidant activity and alpha glucosidase inhibitory potency in these colored grains. The TPC was measured using a Folin-Ciocalteu assay, while the total antioxidant activity was determined by a method based on the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. Among all of the studied colored grains, black rice possessed the highest TPC, which was 86 times greater than that of red rice. In addition, black rice had the highest total anthocyanin contents and alpha glucosidase inhibitory activity. A significant positive correlation of the antioxidant activity and alpha-glucosidase inhibitory activity with total anthocyanin content and TPC was observed in this study. It is concluded that black rice possesses the highest antioxidant activity and alpha-glucosidase inhibitory among all of the colored grains tested and can be further explored as a functional food. PMID- 19904936 TI - Molecular weights of cyclic and hollow clusters measured by DOSY NMR spectroscopy. AB - The Stokes-Einstein expression of the diffusion coefficient as a function of the hydrodynamic radius of the diffusing object does not explicitly carry the mass dependency of the object. It is possible to correlate the translational self diffusion coefficients D with the molecular weight M for an ensemble of cyclic or hollow clusters ranging from about 200 to 30,000 g x mol(-1). From this correlation, the mass of a cluster can be deduced from its diffusion coefficient. Consistency of diffusion as a power law of mass and Stokes-Einstein formulation is completely fulfilled with the selected compounds of this contribution. PMID- 19904937 TI - Tea catechins induce the conversion of preformed lysozyme amyloid fibrils to amorphous aggregates. AB - Natural polyphenols are major constituents of plant foods and herbs. Numerous studies have demonstrated that natural polyphenols inhibited amyloid formation and destabilized the preformed amyloid fibrils. However, the molecular mechanism for the antiamyloidogenesis of polyphenols is still unclear and remains to be further explored. In the present study, the preformed lysozyme fibrils were used as an in vitro model to study the disruptive effects of tea catechins on amyloid fibrils. Results showed that tea catechins induced the conversion of lysozyme fibrils to amorphous aggregates and inhibited fibril-induced hemolysis. Hydroquinone also showed disruptive effect on the fibrils, whereas phenol and two typical antioxidants, ascorbic acid and alpha-tocopherol, did not affect the fibrillar structure, suggesting that polyphenolic structure is essential for fibril deposition. Correlation analyses indicate that the fibril-depositing effects were related to both the antioxidative potency and hydrophobicity of tea catechins. These findings provide new evidence for comprehensive understanding of the interaction between natural polyphenols and amyloid fibrils. PMID- 19904938 TI - Buckyballs meet viral nanoparticles: candidates for biomedicine. AB - Fullerenes such as C(60) show promise as functional components in several emerging technologies. For biomedical applications, C(60) has been used in gene- and drug-delivery vectors, as imaging agents, and as photosensitizers in cancer therapy. A major drawback of C(60) for bioapplications is its insolubility in water. To overcome this limitation, we covalently attached C(60) derivatives to Cowpea mosaic virus and bacteriophage Qbeta virus-like particles, which are examples of naturally occurring viral nanoparticle (VNP) structures that have been shown to be promising candidates for biomedicine. Two different labeling strategies were employed, giving rise to water-soluble, stable VNP-C(60) and VNP PEG-C(60) conjugates. Samples were characterized using a combination of transmission electron microscopy, scanning transmission electron microscopy (STEM), gel electrophoresis, size-exclusion chromatography, dynamic light scattering, and Western blotting. "Click" chemistry bioconjugation using a poly(ethylene glycol) (PEG)-modified propargyl-O-PEG-C(60) derivative gave rise to high loadings of fullerene on the VNP surface, as indicated by the imaging of individual C(60) units using STEM. The cellular uptake of dye-labeled VNP-PEG C(60) complexes in a human cancer cell line was found by confocal microscopy to be robust, showing that cell internalization was not inhibited by the attached C(60) units. These results open the door for the development of novel therapeutic devices with potential applications in photoactivated tumor therapy. PMID- 19904939 TI - Development and evaluation of granule and emulsifiable concentrate formulations containing Derris elliptica extract for crop pest control. AB - Derris elliptica Benth. extracts containing rotenone have long been used as natural insecticides, but time-consuming preparation processes and the short shelf life of the extract limit their use in pest control. In this study, stable water-dispersible granules and emulsifiable concentrate liquids containing Derris extract (equivalent to 5% w/w of rotenone) were developed with simple techniques. Accelerated degradation kinetics of rotenone in the Derris extract, and in both formulations, indicated that its degradation followed first-order kinetics. The predicted half-life (t(1/2)) and shelf life (t(90%)) at 30 degrees C of rotenone in Derris extract were 520 and 79 days, respectively. Derris granules and emulsifiable concentrate clearly prolong the stability of rotenone 8-fold (t(90%) = 633 days) and 1.4-fold (t(90%) = 110 days), respectively. The study of rotenone degradation after application onto plants indicated that both formulations would be effective for up to 3 days after spraying. Preliminary efficacy testing indicated that the Derris emulsifiable concentrate was clearly more effective than Derris water-dispersible granules in controlling Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae). PMID- 19904940 TI - Controlling the specificity of modularly assembled small molecules for RNA via ligand module spacing: targeting the RNAs that cause myotonic muscular dystrophy. AB - Myotonic muscular dystrophy types 1 and 2 (DM1 and DM2, respectively) are caused by expansions of repeating nucleotides in noncoding regions of RNA. In DM1, the expansion is an rCUG triplet repeat, whereas the DM2 expansion is an rCCUG quadruplet repeat. Both RNAs fold into hairpin structures with periodically repeating internal loops separated by two 5'GC/3'CG base pairs. The sizes of the loops, however, are different: the DM1 repeat forms 1 x 1 nucleotide UU loops while the DM2 repeat forms 2 x 2 nucleotide 5'CU/3'UC loops. DM is caused when the expanded repeats bind the RNA splicing regulator Muscleblind-like 1 protein (MBNL1), thus compromising its function. Therefore, one potential therapeutic strategy for these diseases is to prevent MBNL1 from binding the toxic RNA repeats. Previously, we designed nanomolar inhibitors of the DM2-MBNL1 interaction by modularly assembling 6'-N-5-hexyonate kanamycin A (K) onto a peptoid backbone. The K ligand binds the 2 x 2 pyrimidine-rich internal loops found in the DM2 RNA with high affinity. The best compound identified from that study contains three K modules separated by four propylamine spacing modules and is 20-fold selective for the DM2 RNA over the DM1 RNA. Because the modularly assembled K-containing compounds also bound the DM1 RNA, albeit with lower affinity, and because the loop size is different, we hypothesized that the optimal DM1 RNA binder may display K modules separated by a shorter distance. Indeed, here the ideal DM1 RNA binder has only two propylamine spacing modules separating the K ligands. Peptoids displaying three and four K modules on a peptoid scaffold bind the DM1 RNA with K(d)'s of 20 nM (3-fold selective for DM1 over DM2) and 4 nM (6-fold selective) and inhibit the RNA-protein interaction with IC(50)'s of 40 and 7 nM, respectively. Importantly, by coupling the two studies together, we have determined that appropriate spacing can affect binding selectivity by 60-fold (20- x 3-fold). The trimer and tetramer also bind approximately 13- and approximately 63-fold more tightly to DM1 RNAs than does MBNL1. The modularly assembled compounds are cell permeable and nontoxic as determined by flow cytometry. The results establish that for these two systems: (i) a programmable modular assembly approach can provide synthetic ligands for RNA with affinities and specificities that exceed those of natural proteins; and, (ii) the spacing of ligand modules can be used to tune specificity for one RNA target over another. PMID- 19904941 TI - Analysis of free fatty acids in beer: comparison of solid-phase extraction, solid phase microextraction, and stir bar sorptive extraction. AB - Solid-phase extraction (SPE), solid-phase microextraction (SPME) using carbowax/divinylbenzen fiber, and stir bar sorptive extraction (SBSE) followed by solvent back extraction have been used for the extraction of free fatty acids (caproic, caprylic, pelargonic, capric, lauric, myristic, palmitic, stearic, oleic, linoleic, and linolenic acids) from beer. Subsequent gas chromatographic analyses with flame ionization detection were used for the determination of these compounds. Medium-chain fatty acids (caproic-lauric acid) were determined as free acids, and long-chain fatty acids (myristic-linolenic acids) were determined as methyl esters after methylation by BF(3)-methanol 14%. Linearity, recovery, and repeatability of all methods have been determined and compared with the SPE method used as a reference (SPME method was used only for medium-chain fatty acid determination). All three procedures provide similar working parameters characterized by high repeatability (2.3-16.3%) and good linearity (correlation coefficient ranging from 0.9919 to 0.9999). Results of beer analyses obtained by using these three methods were highly correlated. Although all methods provide compatible alternatives, for medium-chain fatty acid analysis SPME may be a more appropriate technique due to its operational simplicity, repeatability, and low cost. PMID- 19904942 TI - Protonation-dissociation reactions of imazamethabenz-methyl and imazamethabenz Acid in relation to their soil sorption and abiotic degradation. AB - This paper present ultraviolet-visible absorption spectra of imazamethabenz methyl (IMBM) (mixture of the isomers methyl 6-[(RS)-4-isopropyl-4-methyl-5-oxo-2 imidazolin-2-yl]-m-toluate, m-imazamethabenz, and methyl 2-[(RS)-4-isopropyl-4 methyl-5-oxo-2-imidazolin-2-yl]-p-toluate, p-imazamethabenz) and the corresponding carboxylic acid, imazamethabenz-acid (IMBA). The spectral characteristics are determined as functions of the pH. The appreciable absorbance in the visible (or near-ultraviolet) region of the spectra indicates that the natural photolytic degradation is possible. From variations of the maximum absorbances of the bands, the pK values of 3.4 +/- 0.2 and 9.4 +/- 0.2 were obtained for protonation of the imidazol (=N-) nitrogen and dissociation of the NH imidazol nitrogen of IMBM, respectively. For IMBA, the dissociation pK of the carboxylic group is very close to that of the imidazol (=N-) nitrogen, both being close to 3.3. The dissociation pK of the NH imidazol nitrogen for IMBA is 9.6 +/- 0.2. The role of the acid-base reactions on the sorption on soils of these herbicides is discussed. PMID- 19904943 TI - Modular synthesis of heparan sulfate oligosaccharides for structure-activity relationship studies. AB - Although hundreds of heparan sulfate binding proteins have been identified and implicated in a myriad of physiological and pathological processes, very little information is known about the ligand requirements for binding and mediating biological activities by these proteins. This difficulty results from a lack of technology for establishing structure-activity relationships, which in turn is due to the structural complexity of natural heparan sulfate (HS) and difficulties of preparing well-defined HS oligosaccharides. To address this deficiency, we developed a modular approach for the parallel combinatorial synthesis of HS oligosaccharides that utilizes a relatively small number of selectively protected disaccharide building blocks, which can easily be converted into glycosyl donors and acceptors. The utility of the modular building blocks has been demonstrated by the preparation of a library of 12 oligosaccharides, which has been employed to probe the structural features of HS for inhibiting the protease, BACE-1. The complex variations in activity with structural changes support the view that important functional information is embedded in HS sequences. Furthermore, the most active derivative provides an attractive lead compound for the preparation of more potent compounds, which may find use as a therapeutic agent for Alzheimer's disease. PMID- 19904945 TI - Reaction of Br2 with adsorbed CO on Pt, studied by the surface interrogation mode of scanning electrochemical microscopy. AB - Scanning electrochemical microscopy surface interrogation (SI-SECM) in the cyclic voltammetry mode was successfully used to detect and quantify adsorbed CO on a Pt electrode by reaction with electrogenerated Br(2). The two-electrode setup used in this new technique allowed the production of Br(2) on an interrogator tip, which reported a transient positive feedback above a Pt substrate at open circuit as an indication of the reactivity of this halogen with CO((ads)). Br(-) and CO(2) are shown to be the main products of the reaction (in the absence of O(2)), which may involve the formation of bromophosgene as a hydrolyzable intermediate. Under saturation conditions, CO((ads)) was reproducibly quantified at the polycrystalline Pt surface with theta(CO) approximately = 0.5. The reaction is shown to be blocked by the action of pre-adsorbed cyanide, which demonstrates the surface character of the process. The formation of CO(2) as an end product was further tested in a bulk experiment: addition of Pt black to a mixture of Br(2) in 0.5 M H(2)SO(4) through which CO was bubbled gave a precipitate of BaCO(3) in a saturated solution of Ba(OH)(2). The use of SI-SECM allowed access to a reaction that would otherwise be difficult to prove through conventional electrochemistry on a single electrode. PMID- 19904944 TI - Co-adsorption and separation of CO2-CH4 mixtures in the highly flexible MIL 53(Cr) MOF. AB - The present study attempts to understand the use of the flexible porous chromium terephthalate Cr(OH)(O(2)C-C(6)H(4)-CO(2)) denoted MIL-53(Cr) (MIL = Material from Institut Lavoisier) for the separation of mixtures of CO(2) and CH(4) at ambient temperature. The coadsorption of CO(2) and CH(4) was studied by a variety of different techniques. In situ synchrotron X-ray Powder Diffraction allowed study of the breathing of the solid upon adsorption of the gas mixtures and simultaneously measured Raman spectra yielded an estimation of the adsorbed quantities of CO(2) and CH(4), as well as a quantification of the fraction of the narrow pore (NP) and the large pore (LP) form of MIL-53. Quantitative coadsorption data were then measured by gravimetry and by breakthrough curves. In addition, computer simulation was performed to calculate the composition of the adsorbed phase in comparison with experimental equilibrium isotherms and breakthrough results. The body of results shows that the coadsorption of CO(2) and CH(4) leads to a similar breathing of MIL-53(Cr) as with pure CO(2). The breathing is mainly controlled by the partial pressure of CO(2), but increasing the CH(4) content progressively decreases the transformation of LP to NP. CH(4) seems to be excluded from the NP form, which is filled exclusively by CO(2) molecules. The consequences in terms of CO(2)/CH(4) selectivity and the possible use of MIL-53(Cr) in a PSA process are discussed. PMID- 19904946 TI - Low-temperature addition of organolithiums to functionalized vinylsilanes under formation of secondary alpha-lithiated alkylsilanes. AB - At -78 degrees C and in apolar solvents, the presence and quality of a coordinating side arm in vinylsilanes dictates both their reactivity toward alkyllithiums and the structural motif in the solid state, deciding whether a defined carbolithiation, slow polymerization or no reaction at all occurs. The crystal structures of the carbolithiation products of two vinylsilanes and methyllithium are presented, and the energetic barriers of the addition step with different alkyllithiums are investigated. PMID- 19904947 TI - Predicting the structure of supramolecular dendrimers via the analysis of libraries of AB3 and constitutional isomeric AB2 biphenylpropyl ether self assembling dendrons. AB - The synthesis of 4'-hydroxy-4-biphenylpropionic, 3',4'-dihydroxy-4 biphenylpropionic, 3',5'-dihydroxy-4-biphenylpropionic, and 3',4',5'-trihydroxy-4 biphenylpropionic methyl esters via three efficient and modular strategies including one based on Ni-catalyzed borylation and sequential cross-coupling is reported. These building blocks were employed in a convergent iterative approach to the synthesis of one library of 3,4,5-trisubstituted and two libraries of constitutional isomeric 3,4- and 3,5-disubstituted biphenylpropyl ether dendrons. Structural and retrostructural analysis of supramolecular dendrimers revealed that biphenylpropyl ether dendrons self-assemble and self-organize into the same periodic lattices and quasi-periodic arrays observed in previously reported libraries, but with larger dimensions, different mechanisms of self-assembly, and improved solubility, thermal, acidic, and oxidative stability. The different mechanisms of self-assembly led to the discovery of two new supramolecular structures. The first represents a new banana-like lamellar crystal with a four layer repeat. The second is a giant vesicular sphere self-assembled from 770 dendrons that exhibits an ultrahigh molar mass of 1.73 x 10(6) g/mol. Thus, the enhanced size of the self-assembled structures constructed from biphenylpropyl ether dendrons permitted for the first time discrimination of various molecular mechanisms of spherical self-assembly and elaborated a continuum between small filled spheres and very large hollow spheres that is dictated by the primary structure of the dendron. The comparative analysis of libraries of biphenylpropyl ether dendrons with the previously reported libraries of benzyl-, phenylpropyl-, and biphenyl-4-methyl ether dendrons demonstrated biomimetic self-assembly wherein the primary structure of the dendron and to a lesser extent the structure of its repeat unit determines the supramolecular tertiary structure. A "nanoperiodic table" of self-assembling dendrons and supramolecular dendrimers that allows the prediction of the general features of tertiary structures from primary structures was elaborated. PMID- 19904948 TI - Deactivation pathways of an isolated green fluorescent protein model chromophore studied by electronic action spectroscopy. AB - The mechanism of fluorescence and fluorescence quenching of the green fluorescent protein (GFP) is not well-understood. To gain insight into the effect of the surrounding protein on the chromophore buried at its center, the intrinsic electronic absorption and deactivation pathways of a gaseous model chromophore, p hydroxybenzylidene-2,3-dimethylimidazolone (HBDI) were investigated. No fluorescence from photoactivated gaseous HBDI(-) was detected in the range 480 1100 nm, in line with the ultrafast rate of internal conversion of HBDI(-) in solution. Two different gas-phase deactivation pathways were found: photofragmentation and electron photodetachment. Electronic action spectra for each deactivation pathway were constructed by monitoring the disappearance of HBDI(-) and appearance of product ions as a function of excitation wavelength. The action spectra measured for each pathway are distinct, with electron photodetachment being strongly favored at higher photon energies. The combined (total) gas-phase action spectrum has a band origin at 482.5 nm (23340 cm(-1)) and covers a broad spectral range, 390-510 nm. This extended gas-phase action spectrum exhibits vibronic activity that matches well with the results of previous cold condensed-phase experiments and high-level in vacuo computations, with features evident at +550, +1500, and +2800 cm(-1) with respect to the band origin. PMID- 19904949 TI - Differential sensing using proteins: exploiting the cross-reactivity of serum albumin to pattern individual terpenes and terpenes in perfume. AB - There has been a growing interest in the use of differential sensing for analyte classification. In an effort to mimic the mammalian senses of taste and smell, which utilize protein-based receptors, we have introduced serum albumins as nonselective receptors for recognition of small hydrophobic molecules. Herein, we employ a sensing ensemble consisting of serum albumins, a hydrophobic fluorescent indicator (PRODAN), and a hydrophobic additive (deoxycholate) to detect terpenes. With the aid of linear discriminant analysis, we successfully applied our system to differentiate five terpenes. We then extended our terpene analysis and utilized our sensing ensemble for terpene discrimination within the complex mixtures found in perfume. PMID- 19904950 TI - Controlled release and entrapment of enantiomers in self-assembling scaffolds composed of beta-sheet peptides. AB - As a first step toward utilizing self-assembling peptide scaffolds to create tunable matrices for drug delivery, peptide RADAFI and RADAFII, containing the same amino acid composition but different positions of one phenylalanine residue, scaffolds were prepared for controlled release of chiral enantiomers. The release behaviors depended on the network nanostructures and the guest chirality and were well tailored via loading different amounts of guests. This contribution addressed the relationships among the peptide sequence, the network nanoarchitecture, and the controlled release. RADAFII systems provided a means of controlling the release kinetics for L- and D-phenylalanine, which was achieved through the facile pi-pi stacking between the aromatic rings of L-isomer and those in RADAFII sequence and through the appropriate scaffold nanoarchitecture. The concept of controlled release for enantiomers via dominating the network nanostructures can also be harnessed in the de novo design of delivery systems with specific structural features for some special biomolecules. PMID- 19904951 TI - Evidence of mineralization activity and supramolecular assembly by the N-terminal sequence of ACCBP, a biomineralization protein that is homologous to the acetylcholine binding protein family. AB - Several biomineralization proteins that exhibit intrinsic disorder also possess sequence regions that are homologous to nonmineral associated folded proteins. One such protein is the amorphous calcium carbonate binding protein (ACCBP), one of several proteins that regulate the formation of the oyster shell and exhibit 30% conserved sequence identity to the acetylcholine binding protein sequences. To gain a better understanding of the ACCBP protein, we utilized bioinformatic approaches to identify the location of disordered and folded regions within this protein. In addition, we synthesized a 50 AA polypeptide, ACCN, representing the N-terminal domain of the mature processed ACCBP protein. We then utilized this polypeptide to determine the mineralization activity and qualitative structure of the N-terminal region of ACCBP. Our bioinformatic studies indicate that ACCBP consists of a ten-stranded beta-sandwich structure that includes short disordered sequence blocks, two of which reside within the primarily helical and surface accessible ACCN sequence. Circular dichroism studies reveal that ACCN is partially disordered in solution; however, ACCN can be induced to fold into an alpha helix in the presence of TFE. Furthermore, we confirm that the ACCN sequence is multifunctional; this sequence promotes radial calcite polycrystal growth on Kevlar threads and forms supramolecular assemblies in solution that contain amorphous-appearing deposits. We conclude that the partially disordered ACCN sequence is a putative site for mineralization activity within the ACCBP protein and that the presence of short disordered sequence regions within the ACCBP fold are essential for function. PMID- 19904952 TI - Binding of beta-lactoglobulin to pectins varying in their overall and local charge density. AB - The formation of complexes between proteins and polysaccharides is of great importance for many food systems like foams, emulsions, acidified milk drinks, and so on. The complex formation between beta-lactoglobulin (beta-lg) and pectins with a well-defined physicochemical fine structure has been studied to elucidate the influence of overall charge and local charge density of pectin on the complex formation. Binding isotherms of beta-lg to pectin are constructed using fluorescence anisotropy, which is shown to be an excellent technique for this purpose, as it is fast and requires low sample volumes. From the binding isotherms the maximal adsorbed amount, binding constant (k(obs)) and the cooperativity of binding are obtained at different ionic strengths. The Hill model is used to fit the binding isotherms and is shown to be preferable over a Langmuir fit. At pH 4.25, k(obs) shows a maximum at an ionic strength of 10 mM when using a low methyl esterified pectin (LMP) due to the balance of attractive and repulsive electrostatic forces between beta-lg and pectin and beta-lg neighbors. For two high methyl esterified pectins, one with a blockwise distribution of methyl esters (HMP(B)) and one with a random distribution (HMP(R)), this ionic strength maximum is absent and k(obs) decreases with increasing ionic strength. k(obs) is found to be largest for LMP and HMP(B) and considerably lower for HMP(R). A positive cooperativity is observed for both LMP (above an ionic strength of 45 mM) and HMP(R) (above an ionic strength of 15 mM) but not for HMP(B). Positive cooperativity is thought to be caused by a rearrangement of the pectin helix structure caused by binding of beta-lg, thus creating new or binding sites with a higher affinity. To attain strong binding of beta-lg to pectin it is preferable to use a pectin with a blockwise distribution of methyl esters. When complex formation takes place in high ionic strength media an LMP gives the best results, while at low ionic strength a high methyl esterified pectin with blockwise distribution may give better results, due to reduced electrostatic repulsion between both pectin and beta-lg and beta-lg neighbors. PMID- 19904953 TI - Aqueous electrodeposition of Ge monolayers. AB - The electrodeposition of germanium on Au(111) in aqueous solutions has been investigated by means of cyclic voltammetry, Auger electron spectroscopy, and in situ scanning tunneling microscopy (STM). The data yield a picture of germanium deposition, which starts with the formation of two well-ordered hydroxide phases, with 1/3 ML and 4/9 ML coverages upon initial reduction of the Ge(IV) species (probably H(2)GeO(3) at pH 4.7). Those structures appear to result from a three electron reduction to form surface-limited structures with (square root(3) x square root(3))R30 degrees or (3 x 3) unit cells, respectively. Further reduction, probably in a two-electron process from the hydroxide structures, resulted in a germanium hydride structure, again surface-limited, with a coverage of close to 0.8 ML. The hydride structure is very flat, though with the periodic modulation characteristic of a Moire pattern. Longer deposition times and lower potentials resulted in increased coverage of Ge in some cases, but with apparently limited coverage as a function of pH. The maximum Ge coverage, about 4 ML, was observed using a pH 9.32 deposition solution. At potentials negative of the Moire pattern, about -850 mV versus Ag/AgCl, a "corruption" of the smooth Moire pattern occurred. This roughening appears to mark the initial formation of a Au-Ge alloy, accounting for the observation of coverage in excess of that needed to form the Moire pattern at some pH values. PMID- 19904954 TI - Synthesis and evaluation of phenylalanine-modified hyperbranched poly(amido amine)s as promising gene carriers. AB - Hyperbranched poly(amido amine) (HPAMAM), which is structurally analogous to PAMAM dendrimers, has been proposed to be an effective agent for gene delivery. The facile synthesis of HPAMAM with scalable productivity by one-pot polymerization of monomers of methyl acrylate (MA) and diethylenetriamine (DETA) has been set up previously. In this study, the HPAMAM was further modified on the terminal amino groups with phenylalanine to various degrees (HPAMAM-PHE30, PHE45, PHE60). We showed that HPAMAM and HPAMAM-PHEs were all able to form complexes with plasmid DNA (pDNA) at various mass ratios. The cytotoxicity and transfection efficiencies of these polymers were evaluated in SMMC-7721 and COS-7 cell lines. The PHE modifications affected the cell transfection efficiency significantly. The HPAMAM-PHE60 was the most efficient, with transfection activities consistently higher than the commercial transfection reagent PEI. Our study demonstrated that HPAMAM-PHEs may be good new materials for gene delivery and other applications because of its large-scale availability, economical cost, and low toxicity. PMID- 19904955 TI - Long time spreading of a microdroplet on a smooth solid surface. AB - The long time spreading of microdroplets on a smooth solid surface is studied experimentally. An empirical expression is obtained for the spreading area as a function of time showing a final area when spreading stops. The mean film thickness of this final area appears to be independent of the initial volume of the droplet and of the spreading dynamics. A theoretical model is developed to predict this final uniform film thickness, based on volume conservation and the principle of minimum energy. Good agreement is found between the theoretical and experimental results. PMID- 19904956 TI - Selective formation and structural properties of rhombic dodecahedral [70]fullerene microparticles formed by reaction with aliphatic diamines. AB - We have accomplished the selective formation of rhombic dodecahedral microparticles on the submicrometer to micrometer scale by the reaction of [70]fullerene (C(70)) with primary aliphatic diamines. The morphology of the resultant microparticles was analyzed by scanning electron microscopy, powder X ray diffraction, and other spectroscopic methods, demonstrating that the resultant particles held a rhombic dodecahedral shape having a simple cubic lattice structure and that primary aliphatic amines were mostly trapped inside the particles through electronic interaction between C(70) and amines. Furthermore, we have discovered interesting structural characteristics in which the incorporated amines could be removed from the C(70) microparticles or exchanged with other primary aliphatic diamines. PMID- 19904957 TI - Hydration of lysozyme: the protein-protein interface and the enthalpy-entropy compensation. AB - Water sorption isotherms of proteins are usually interpreted with such models as BET or GAB that imply the formation of multilayers at solid-gas interface. However, this approach is not applicable to globular proteins such as humid lysozyme where a solid-gas interface does not exist. Another popular approach is the D'Arcy-Watt model, where besides the formation of multilayers the heterogeneity of energies of sorption sites of proteins is taken into account. Here we present sorption calorimetric data on the hydration of lysozyme that confirms the existence of the heterogeneity. The magnitude of the heterogeneity is, however, lower than one can expect on the basis of the existence of a solid gas interface. Moreover, the calorimetric data show a strong enthalpy-entropy compensation that leads to almost constant effective free energy of hydration in the activity range normally used for fitting the data to sorption models. This allows the use of the Langmuir equation for the fitting of the initial part of the sorption isotherm of lysozyme. Assuming the formation of a monolayer of water at the protein-protein interface, one can estimate the size of the lysozyme molecules from the sorption isotherm. The result of this estimation is in good agreement with the structural data on lysozyme, which supports the presented approach. PMID- 19904958 TI - Homopairings of the artificial nucleobase 1H-benzoimidazole-4,7-dione. AB - All planar homopairings of the artificial nucleobase 1H-benzoimidazole-4,7-dione are reported for the first time in this study. Using the idea of binding sites discussed in our previous work and an ab initio density functional theory method we predict 13 homopairs. The stabilization energies of the homopairs vary from 0.13 to -0.69 eV. The collected data on all the planar homopairs reported here may be useful when constructing assemblies of this artificial base on various solid substrates. PMID- 19904959 TI - Peroxyl-radical-scavenging activity of garlic: 2-propenesulfenic acid versus allicin. AB - The OOH radical reactions with allicin and its Cope elimination products (2 propenesulfenic acid and thioacrolein) in aqueous solution have been studied. The CBS-QB3 quantum chemistry method has been used, with geometries and frequencies at BHandHLYP/6-311++G(d,p) level and conventional transition state theory. 2 Propenesulfenic acid is predicted to be over 1000 times more reactive toward OOH radical than allicin (2.60 x 10(7) vs 7.38 x 10(3) L mol(-1) s(-1), at 298 K). Accordingly, our results strongly support the novel suggestion by Vaidya et al. (Angew. Chem., Int. Ed. 2009, 48, 157) that the active ingredient responsible for the free radical scavenging activity of garlic is actually 2-propenesulfenic acid and not allicin. In addition, direct reaction branching ratios and product distribution for the three studied reactions are proposed for the first time. PMID- 19904960 TI - Palladium-catalyzed 2-arylation of pyrroles. AB - A methodology that affords N-alkyl-2-arylpyrroles and N-aryl-2-arylpyrroles via direct coupling from aryl iodides has been developed. After examining various reaction parameters: solvent, ratio of reagents, catalyst, base and additives the optimal conditions for the condensation were identified. Two crucial factors, (a) anhydrous DMSO as solvent and (b) 5 M excess of pyrrole counterpart, were found to strongly influence the reaction outcome. The conditions identified (PdCl(2)(PPh(3))(2), AgOAc, anhyd DMSO, KF, 100 degrees C, 5 h) resulted in the formation of 2-arylpyrroles in 14-80% yield. Furthermore, the synthesis is compatible with electron-withdrawing and electron-donating groups on the aryl moiety. PMID- 19904962 TI - Slow solvation dynamics in the microheterogeneous water channels of nafion membranes. AB - Solvation dynamics in Nafion membrane is studied using the well-known solvation probe, coumarin 102 (C102). In native Nafion membrane, the fluorescence maximum of C102 occurs at 525 nm. The decays recorded at different wavelengths are superimposable. There is no time-dependent Stokes shift (TDSS) in the time scale of the experiment. This is rationalized in light of the strongly acidic environment in Nafion membrane, which causes the C102 molecules to become protonated. The protonated molecules are bound tightly to the negatively charged sulfonate groups. In Na(+)- and Me(4)N(+)-exchanged Nafion membranes, the fluorescence gets blue-shifted by 65 nm, indicating the deprotonation of the cation and formation of neutral C102 in these membranes. TDSS is observed in the picosecond-nanosecond time scale, in the cation-exchanged Nafion membranes, although the amount of Stokes' shift is rather small, as compared to that observed in organic solvents, indicating that a significant amount of the solvation is ultrafast and is missed in the present experiment. The observed solvation dynamics is bimodal with fast ( approximately 1 ns) and slow (>10 ns) components. The ultraslow component is ascribed to the quasi-static water molecules in the Nafion membrane. The difference in the extents of apparently missing ultrafast components, between Me(4)N(+)- and Na(+)-substituted membranes is rationalized by a model involving the difference in distributions of the cations in the water channel. PMID- 19904961 TI - Lanthanide oleates: chelation, self-assembly, and exemplification of ordered nanostructured colloidal contrast agents for medical imaging. AB - Eight lanthanide(III) oleates have been prepared and characterized. The chelation and self-assembly structures of these rare-earth oleates have been studied by elemental analysis, Fourier transfer infrared spectroscopy (FTIR), and X-ray powder diffraction (XRD) analysis. Elemental analysis and FTIR results indicate that three oleate anions are complexed with one lanthanide cation and, with the exception of anhydrous cerium(III) oleate, form either a mono- or a hemihydrate. The X-ray analysis showed that the neat lanthanide soaps have a lamellar bilayer structure at room temperature. The thermal behavior has been investigated by cross-polarized optical microscopy (POM), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). POM scans showed that all the lanthanide oleates form a lamellar phase in the presence of excess water. Small angle X-ray scattering (SAXS) and XRD were used to investigate the internal structure of the bulk lanthanide oleates in excess water, and these X-ray results confirmed that the lanthanide oleates do not swell in water. Select lanthanide oleates were dispersed in water to form nonswelling lamellar submicrometer particles, confirmed by dynamic light scattering (DLS) and synchrotron SAXS measurements. NMR results indicated that colloidal dispersions of lanthanide oleates containing paramagnetic ions, such as gadolinium(III), terbium(III), and dysprosium(III), have a significant effect on the longitudinal (T(1)) and transverse (T(2)) relaxation times of protons in water. Time-resolved fluorescence measurements have demonstrated that colloidal dispersions of europium(III) oleate exhibit strong luminescence. The rare earth metal soaps exemplify the potential of self-assembled chelating amphiphiles as contrast agents in medical imaging modalities such as magnetic resonance imaging (MRI) and fluorescence imaging. PMID- 19904963 TI - Vibrational spectroscopy of secondary amine salts: 1. Assignment of NH(2)(+) stretching frequencies in crystalline phases. AB - The NH(2)(+) stretching modes of secondary amine salts have been previously studied, but the band assignments are inconsistent between the various studies. This paper assigns characteristic NH(2)(+) group frequencies between approximately 2500 and 2400 cm(-1). Crystal structures of four diamine salts are reported here. Vibrational frequencies were calculated with the B3LYP hybrid Hartree-Fock/density functional method and the 6-31G(d) split-valence plus polarization basis set, and the results are in agreement with the experimental frequencies. Deuterium dilution experiments result in a group of sharply featured bands between the NH(2)(+) and the ND(2)(+) stretching bands. These bands, located between 2200 and 2100 cm(-1), are attributed to modes that contain contributions from coupled N-H and N-D stretching motions. PMID- 19904964 TI - Detection of in vivo matrix metalloproteinase activity using microdialysis sampling and liquid chromatography/mass spectrometry. AB - Matrix metalloproteinases (MMPs) are a family of endoproteases that break down extracellular matrix and whose upregulation contributes to several diseases. A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed to quantify MMP-1 and MMP-9 substrates and their N-terminal peptide products in samples obtained from implanted microdialysis sampling probes. In vitro studies with purified human MMP-1 and MMP-9 were used to optimize the assay and determine the effectiveness of the local delivery of a broad-spectrum MMP inhibitor, GM 6001. Localized delivery of GM 6001 at 10 microM was sufficient to completely inhibit product formation in vitro. In vivo studies in male Sprague-Dawley rats were performed with microdialysis probes implanted into the subcutaneous tissue. Directly after microdialysis probe implantation, infusions of the MMP-1 and MMP-9 substrates (50 microM each) resulted in recovered product concentrations of approximately 2 microM. During a 50 microM GM 6001 coinfusion with the substrates, a 30% and 25% reduction in product formation for the MMP-1 and MMP-9 substrates was obtained, respectively. Blank dialysates were negative for enzymatic activity that could cleave the MMP substrates. This method allowed for the activity of different MMPs surrounding the microdialysis probe to be observed during in vivo sampling. PMID- 19904966 TI - Malonamide-functionalized gold nanoparticles for selective, colorimetric sensing of trivalent lanthanide ions. AB - A selective and sensitive molecular sensor for trivalent lanthanide (Ln(3+)) ions based upon a malonamide-functionalized gold nanoparticle was developed for colorimetric detection in water. A new synthetic approach permits nanoparticle synthesis, stabilization, and incorporation of a selective lanthanide binding site in a single, direct step. The design incorporates a specifically tailored dual function precursor ligand that bears a sodium thiosulfate (Bunte salt) group that links to the gold nanoparticle core and a tetramethylmalonamide (TMMA) group that serves as a selective Ln(3+) binding site. The sensor's colorimetric response to lanthanide ions is immediate, and it is sensitive down to approximately 50 nM for Eu(3+) and Sm(3+). This study demonstrates a general strategy for direct, convenient nanoparticle synthesis that enables the incorporation of analyte binding groups directly to the nanoparticle surface, allowing colorimetric sensors to be developed for widespread use. The one-step synthesis offers uniform surface ligand composition, reduces the volume of waste generated during nanoparticle synthesis and purification, produces functionalized gold nanoparticles that are stable in nonmodified aqueous environments, and offers colorimetric detection at ambient temperature. PMID- 19904967 TI - Experimental and theoretical investigation of the charge-separation energies of hydrated zinc(II): redefinition of the critical size. AB - In the preceding article, the hydration energies of Zn(2+)(H(2)O)(n) complexes, where n = 6-10, were measured using threshold collision-induced dissociation (CID) in a guided ion beam tandem mass spectrometer (GIBMS) coupled with an electrospray ionization (ESI) source. The present investigation explores the charge-separation processes observed, Zn(2+)(H(2)O)(n) --> ZnOH(+)(H(2)O)(m) + H(+)(H(2)O)(n-m-1), and the competition between this process and the loss of water. Our results demonstrate that charge-separation processes occur at variable complex sizes of n = 6, 7, and 8, prompting a redefinition of the critical size for charge separation. Experimental kinetic energy-dependent cross sections are analyzed to yield 0 K threshold energies for the charge-separation products and the effects of competition with this channel on the energies for losing one and two water ligands after accounting for multiple collisions, kinetic shifts, and energy distributions. A complete reaction coordinate is calculated for the n = 7 complex dissociating into ZnOH(+)(H(2)O)(3) + H(+)(H(2)O)(3). Calculated rate limiting transition states for n = 6-8 are also compared to experimental threshold measurements for the charge-separation processes. PMID- 19904969 TI - Hydration energies of zinc(II): threshold collision-induced dissociation experiments and theoretical studies. AB - The first experimentally determined sequential bond dissociation energies of Zn(2+)(H(2)O)(n) complexes, where n = 6-10, are measured using threshold collision-induced dissociation in a guided ion beam tandem mass spectrometer coupled with an electrospray ionization source. Kinetic energy dependent cross sections are obtained and analyzed to yield 0 K threshold measurements for the loss of one and two water ligands after accounting for multiple collisions, kinetic shifts, and energy distributions. The threshold measurements are then converted from 0 to 298 K values to give the hydration energies for sequentially losing one water from each parent complex. Theoretical geometry optimizations and single-point energy calculations are performed using several levels of theory for comparison to experiment. Although different levels of theory disagree on the ground-state conformation of most complexes examined here leading to potential ambiguities in the final thermochemical values, calculations at the MP2(full) level provide the best agreement with experiment. On this basis, the present experiments are most consistent with the inner solvent shell of Zn(2+) being five waters, except for Zn(2+)(H(2)O)(6) where all waters bind directly to the metal ion. The charge separation process, Zn(2+)(H(2)O)(n) --> ZnOH(+)(H(2)O)(m) + H(+)(H(2)O)(n-m-1), which is in competition with the loss of water from the parent complex, is also observed for n = 6-8. These processes are analyzed in detail in the following paper. PMID- 19904968 TI - The 8 and 5 kDa fragments of plasma gelsolin form amyloid fibrils by a nucleated polymerization mechanism, while the 68 kDa fragment is not amyloidogenic. AB - Familial amyloidosis of Finnish type (FAF), or gelsolin amyloidosis, is a systemic amyloid disease caused by a mutation (D187N/Y) in domain 2 of human plasma gelsolin, resulting in domain 2 misfolding within the secretory pathway. When D187N/Y gelsolin passes through the Golgi, furin endoproteolysis within domain 2 occurs as a consequence of the abnormal conformations that enable furin to bind and cleave, resulting in the secretion of a 68 kDa C-terminal fragment (amino acids 173-755, C68). The C68 fragment is cleaved upon secretion from the cell by membrane type 1 matrix metalloprotease (MT1-MMP), affording the 8 and 5 kDa fragments (amino acids 173-242 and 173-225, respectively) comprising the amyloid fibrils in FAF patients. Herein, we show that the 8 and 5 kDa gelsolin fragments form amyloid fibrils by a nucleated polymerization mechanism. In addition to demonstrating the expected concentration dependence of a nucleated polymerization reaction, the addition of preformed amyloid fibrils, or "seeds", was shown to bypass the requirement for the formation of a high-energy nucleus, accelerating 8 and 5 kDa D187N gelsolin amyloidogenesis. The C68 fragment can form small oligomers, but not amyloid fibrils, even when seeded with preformed 8 kDa fragment plasma gelsolin fibrils. Because the 68 kDa fragment of gelsolin does not form amyloid fibrils in vitro or in a recently published transgenic mouse model of FAF, we propose that administration of an MT1-MMP inhibitor could be an effective strategy for the treatment of FAF. PMID- 19904970 TI - Novel one-pot three component reaction for the synthesis of [2 (alkylsulfanyl)imidazo[1,2-a]pyridin-3-yl](aryl)methanone. AB - A one-pot, three-component reaction between pyridine, phenacyl bromide, and thiocyanate is described. The reaction afforded the corresponding special type of fully substituted imidazo[1,2-a]pyridine derivatives in good yields without using any catalyst or activation. PMID- 19904972 TI - Nonadiabatic dynamics of A-state photodissociation of ammonia: a four-dimensional wave packet study. AB - The nonadiabatic photodissociation dynamics of the A-state ammonia (NH(3)) was investigated using a four-dimensional wave packet model. The branching ratio between the excited NH(2)(A(2)A(1)) and ground NH(2)(X(2)B(1)) products was obtained as a function of energy for photodissociation mediated by several low lying vibrational states in the ground electronic state of NH(3). The calculated results could not fully account for the experimental observations of strong mode specificity in nonadiabatic dynamics but agree qualitatively with a recent trajectory-based coupled-surface study using the same potential energy surfaces. Several possible sources of inaccuracy are discussed. PMID- 19904971 TI - Ultrasound-promoted copper-catalyzed azide-alkyne cycloaddition. PMID- 19904973 TI - Atomic force spectroscopy in biological complex formation: strategies and perspectives. AB - Atomic force spectroscopy has become a widely used technique for investigating forces, energies, and dynamics of biomolecular interactions. These studies provide dissociation kinetic parameters by pulling apart proteins involved in a complex. Biological complexes are studied under near-physiological conditions, without labeling procedures, and are probed one at time, the latter allowing to one obtain results which are not averaged over the ensemble. However, to gain reliable information, some experimental aspects have to be carefully controlled. In particular, the immobilization of molecular partners to AFM tips and supports, required to force the molecular dissociation, plays a crucial role in determining the success of the experiments. To actually resolve single interactions, multiple simultaneous complex dissociations have to be avoided, and nonspecific adhesions, commonly found in these studies, have to be recognized and discarded. This article is aimed at offering a critical revisitation of the atomic force spectroscopy technique applied to the study of biomolecular interactions, highlighting the critical points, identifying strategies to be adopted for a more reliable data extraction and interpretation, and pointing out the experimental and theoretical aspects which still need to be refined. To this purpose, we take advantage of the vast landscape of literature and then proceed into the details of our works. In this respect, we describe the general principles of the technique, the procedures for protein immobilization, and how they can affect the results. We emphasize the use of computational docking to predict molecular complex configurations, when unknown, as a useful approach to select proper anchorage architectures. Additionally, we deal with data acquirement and analysis, with regard to the force curve selection, to the force histograms interpretation, and to the theoretical frameworks used to extract kinetic parameters. Through this, we outline that AFS can be successfully used both to investigate complexes having very different affinities and also to reveal competitive binding mechanisms, thus gaining deeper information about molecular interactions. PMID- 19904974 TI - Redox reactions of Pu(IV) and Pu(III) in the presence of acetohydroxamic acid in HNO(3) solutions. AB - The reduction of Pu(IV) in the presence of acetohydroxamic acid (HAHA) was monitored by vis-NIR spectroscopy. All experiments were performed under low HAHA/Pu(IV) ratios, where only the Pu(IV)-monoacetohydroxamate complex and Pu uncomplexed with HAHA were present in relevant concentrations. Time dependent concentrations of all absorbing species were resolved using molar extinction coefficients for Pu(IV), Pu(III), and the Pu(AHA)(3+) complex by deconvolution of spectra. From fitting of the experimental data by rate equations integrated by a numeric method three reactions were proposed to describe a mechanism responsible for the reduction and oxidation of plutonium in the presence of HAHA and HNO(3). Decomposition of Pu(AHA)(3+) follows a second order reaction mechanism with respect to its own concentration and leads to the formation of Pu(III). At low HAHA concentrations, a two-electron reduction of uncomplexed Pu(IV) with HAHA also occurs. Formed Pu(III) is unstable and slowly reoxidizes back to Pu(IV), which, at the point when all HAHA is decomposed, can be catalyzed by the presence of nitrous acid. PMID- 19904975 TI - Chemical composition of gas- and aerosol-phase products from the photooxidation of naphthalene. AB - The current work focuses on the detailed evolution of the chemical composition of both the gas- and aerosol-phase constituents produced from the OH-initiated photooxidation of naphthalene under low- and high-NO(x) conditions. Under high NO(x) conditions ring-opening products are the primary gas-phase products, suggesting that the mechanism involves dissociation of alkoxy radicals (RO) formed through an RO(2) + NO pathway, or a bicyclic peroxy mechanism. In contrast to the high-NO(x) chemistry, ring-retaining compounds appear to dominate the low NO(x) gas-phase products owing to the RO(2) + HO(2) pathway. We are able to chemically characterize 53-68% of the secondary organic aerosol (SOA) mass. Atomic oxygen-to-carbon (O/C), hydrogen-to-carbon (H/C), and nitrogen-to-carbon (N/C) ratios measured in bulk samples by high-resolution electrospray ionization time-of-flight mass spectrometry (HR-ESI-TOFMS) are the same as the ratios observed with online high-resolution time-of-flight aerosol mass spectrometry (HR ToF-AMS), suggesting that the chemical compositions and oxidation levels found in the chemically-characterized fraction of the particle phase are representative of the bulk aerosol. Oligomers, organosulfates (R-OSO(3)), and other high-molecular weight (MW) products are not observed in either the low- or high-NO(x) SOA; however, in the presence of neutral ammonium sulfate seed aerosol, an organic sulfonic acid (R-SO(3)), characterized as hydroxybenzene sulfonic acid, is observed in naphthalene SOA produced under both high- and low-NO(x) conditions. Acidic compounds and organic peroxides are found to account for a large fraction of the chemically characterized high- and low-NO(x) SOA. We propose that the major gas- and aerosol-phase products observed are generated through the formation and further reaction of 2-formylcinnamaldehyde or a bicyclic peroxy intermediate. The chemical similarity between the laboratory SOA and ambient aerosol collected from Birmingham, Alabama (AL) and Pasadena, California (CA) confirm the importance of PAH oxidation in the formation of aerosol within the urban atmosphere. PMID- 19904976 TI - Combined effects of one 8-hydroxyquinoline/picolinate and "CH"/N substitutions on the geometry, electronic structure and optical properties of mer-Alq(3). AB - With the aim of evaluating the combined effect of one 8-hydroxyquinoline (q)/picolinate (p) and "CH"/N substitutions on the molecular geometry, electronic structure, and optical properties of tris-(8-hydroxyquinoline)aluminum [Alq(3)], the density functional theory (B3LYP) and time-dependent density functional theory (TD-B3LYP), using the 6-31G(d) and 3-21+G(d,p) basis sets were applied on Alq(3), Alq(2)p, and its "CH"/N-substitution derivatives. A comparison of the optimized ground-state (S(0)) geometries has shown that the molecular shape is conserved upon such a substitution. On the basis of the frontier molecular orbital and gap energy (E(g)) calculations, it was shown that, comparatively to the pristine Alq(2)p (and to the original parent Alq(3)), the HOMO and LUMO are stabilized, the net effect being an increasing or a decreasing E(g), depending on the position of the substituted group. The substitution of q(B) by p (from Alq(3) to Alq(2)p) was also found to induce the same feature. Starting from the S(0) and S(1) (first excited state) geometries, the effect of the substitution on the absorption (and emission) spectra was evaluated. It was found that the "CH"/N substitution in different positions on the two 8-hydroxyquinoline ligands may also constitute an efficient approach of tuning the Alq(2)p emitting color. In comparison with both Alq(3) and Alq(2)p, an important blue shift was predicted for the 5-substituted derivative, an important red shift being observed for the 4 substituted one. Also, relatively significant blue and red shifts were predicted for the 7- and 2-substituted derivatives. Finally, revisiting the correlation between the spectrum shifts and the metal-ligand bonding, our recent findings (2) were confirmed. PMID- 19904977 TI - {[Ga(en)3]2(Ge2Te15)}n: a polymeric semiconducting polytelluride with boat-shaped Te8(4-) rings and cross-shaped Te5(6-) units. AB - The reaction of the Zintl compound K(4)Ge(9) with Te and Ga(2)Te(3) in ethylenediamine (en) at 190 degrees C gave the germanium polytelluride {[Ga(en)(3)](2)[(GeTe)(2)(Te(5))(6-)(Te(8))(4-)]}(n) (1). The single-crystal structure analysis revealed that 1 has two different polytelluride fragments: cross-shaped 36-e(-) TeTe(4)(6-) anions and boat-shaped 52-e(-) Te(8)(4-) rings. The new material is a p-type semiconductor at room temperature and switches to n type at 380 K. PMID- 19904978 TI - Phosphorylation of serine 205 by the protein kinase CK2 persists on Pax3-FOXO1, but not Pax3, throughout early myogenic differentiation. AB - The myogenic transcription factor Pax3 plays an essential role in early skeletal muscle development and is a key component in alveolar rhabdomyosarcoma (ARMS), a childhood solid muscle tumor. ARMS is characterized by a t(2;13) chromosomal translocation resulting in the fusion of the 5' Pax3 sequences to the 3' FOXO1 sequences to encode the oncogenic fusion protein, Pax3-FOXO1. Posttranslational modifications such as phosphorylation are common mechanisms by which transcription factors are regulated. Consistent with this fact, we demonstrated in a previous report that Pax3 is phosphorylated on Ser205 in proliferating, but not differentiated, primary myoblasts. However, the kinase that mediates this phosphorylation event has yet to be identified. In addition, it is not known whether Pax3-FOXO1 is phosphorylated at this site or how the phosphorylation of the fusion protein changes during early myogenic differentiation. In this report we identify CK2 (formerly termed "casein kinase II") as the kinase responsible for phosphorylating Pax3 and Pax3-FOXO1 at Ser205 in proliferating mouse primary myoblasts. Furthermore, we demonstrate that, in contrast to wild-type Pax3, phosphorylation at Ser205 persists on Pax3-FOXO1 throughout early myogenic differentiation. Finally, we show that Pax3-FOXO1 is phosphorylated at Ser205 in a variety of translocation-containing ARMS cell lines. The results presented in this report not only suggest a possible mechanism by which the disregulation of Pax3-FOXO1 may contribute to tumorigenesis but also identify a novel target for the development of therapies for the treatment of ARMS. PMID- 19904980 TI - Templated self-assembly of square symmetry arrays from an ABC triblock terpolymer. AB - Self-assembly provides the ability to create well-controlled nanostructures with electronic or chemical functionality and enables the synthesis of a wide range of useful devices. Diblock copolymers self-assemble into periodic arrays of microdomains with feature sizes of typically 10-50 nm, and have been used to make a wide range of devices such as silicon capacitors and transistors, photonic crystals, and patterned magnetic media(1-3). However, the cylindrical or spherical microdomains in diblock copolymers generally form close-packed structures with hexagonal symmetry, limiting their device applications. Here we demonstrate self-assembly of square-symmetry patterns from a triblock terpolymer in which one organometallic block imparts high etch selectivity and etch resistance. Long-range order is imposed on the microdomain arrays by self assembly on topographical substrates, and the orientation of both square lattices and in-plane cylinders is controlled by the substrate chemistry. Pattern transfer is demonstrated by making an array of square-packed 30 nm tall, 20 nm diameter silica pillars. Templated self-assembly of triblock terpolymers can generate nanostructures with geometries that are unattainable from diblock copolymers, significantly enhancing the capabilities of block copolymer lithography. PMID- 19904979 TI - Photo-targeted nanoparticles. AB - We report a novel and simple proof-of-concept of a nanoparticulate system that targets any tissue selectively upon illumination. Nanoparticles were covalently functionalized with the amino acid sequence YIGSR, which adheres to the beta1 integrins present on most cell surfaces. This peptide was masked with a caging group, rendering it biologically inert. Illumination with UV light released the caging group from the YIGSR, allowing binding to cells. PMID- 19904981 TI - Daphhimalenine A, a new alkaloid with an unprecedented skeleton, from Daphniphyllum himalense. AB - Daphhimalenine A (1), a novel alkaloid with a rearrangement C-21 skeleton, containing a unique 1-azabicyclo[5.2.1]decane ring system, was isolated from the leaves of Daphniphyllum himalense, along with biogenetically related alkaloids daphhimalenine B (2) and daphnezomine T. Their structures were established on the basis of spectroscopic data, and the absolute configuration of 1 was assigned by computational methods. A plausible biosynthetic pathway of 1 was also proposed. PMID- 19904982 TI - Evidence of diketopiperazine and oxazolone structures for HA b2+ ion. AB - Peptide fragmentation can lead to an oxazolone or diketopiperazine b(2)(+) ion structure. IRMPD spectroscopy combined with computational modeling and gas-phase H/D exchange was used to study the structure of the b(2)(+) ion from protonated HAAAA. The experimental spectrum of the b(2)(+) ion matches both the experimental spectrum for the protonated cyclic dipeptide HA (a commercial diketopiperazine) and the theoretical spectrum for a diketopiperazine protonated at the imidazole pi nitrogen. A characteristic band at 1875 cm(-1) and increased abundance of the peaks at 1619 and 1683 cm(-1) indicate a second population corresponding to an oxazolone species. H/D exchange also shows a mixture of two populations consistent with a mixture of b(2)(+) ion diketopiperazine and oxazolone structures. PMID- 19904983 TI - Effect of NaCl, myoglobin, Fe(II), and Fe(III) on lipid oxidation of raw and cooked chicken breast and beef loin. AB - Chicken breast and beef loin were ground, and no, NaCl, NaCl+myoglobin, NaCl+Fe(II), or NaCl+Fe(III) additions were made; patties were then prepared. Half of the patties were packaged in oxygen-permeable bags and stored at 4 degrees C for 10 days, and the other half were cooked in a 95 degrees C water bath to an internal temperature of 75 degrees C, packaged in oxygen-permeable zipper bags, and stored at 4 degrees C for 7 days. The oxidative stability of raw and cooked chicken breast and beef loin were determined during storage. Chicken breast was more resistant to various exogenous oxidative factors than beef loin: addition of NaCl did not increase TBARS values and nonheme content of raw chicken breast, but significantly increased those of raw beef loin patties during storage. Addition of NaCl+Mb did not affect lipid oxidation in raw chicken breast patties, but decreased the TBARS of beef loin during storage. Addition of NaCl+Fe(III) or NaCl+Fe(II) increased the TBARS values of both raw chicken breast and beef loin during storage, but the increase was greater in beef loin than in chicken breast. The TBARS values of all cooked chicken breast and beef loin increased during 7 days of storage, but the increases in cooked chicken patties were significantly smaller than those of cooked beef loin patties with the same treatments. Addition of NaCl and cooking caused severe degradation of myoglobin, leading to a significant increase in free ionic iron content in beef loin. It is suggested that free ionic iron is the major catalyst for lipid oxidation, and the low "storage-stable and heat-stable" ferric ion reducing capacity in chicken breast were responsible for the high oxidative stability for raw and cooked chicken breast compared with beef loin under prooxidants, cooking, and storage conditions. PMID- 19904984 TI - Spin-selective charge transport pathways through p-oligophenylene-linked donor bridge-acceptor molecules. AB - A series of donor-bridge-acceptor (D-B-A) triads have been synthesized in which the donor, 3,5-dimethyl-4-(9-anthracenyl)julolidine (DMJ-An), and the acceptor, naphthalene-1,8:4,5-bis(dicarboximide) (NI), are linked by p-oligophenylene (Ph(n)) bridging units (n = 1-5). Photoexcitation of DMJ-An produces DMJ(+*)-An( *) quantitatively, so that An(-*) acts as a high potential electron donor, which rapidly transfers an electron to NI yielding a long-lived spin-coherent radical ion pair (DMJ(+*)-An-Ph(n)-NI(-*)). The charge transfer properties of 1-5 have been studied using transient absorption spectroscopy, magnetic field effects (MFEs) on radical pair and triplet yields, and time-resolved electron paramagnetic resonance (TREPR) spectroscopy. The charge separation (CS) and recombination (CR) reactions exhibit exponential distance dependencies with damping coefficients of beta = 0.35 A(-1) and 0.34 A(-1), respectively. Based on these data, a change in mechanism from superexchange to hopping was not observed for either process in this system. However, the CR reaction is spin-selective and produces the singlet ground state and both (3*)An and (3*)NI. A kinetic analysis of the MFE data shows that superexchange dominates both pathways with beta = 0.48 A(-1) for the singlet CR pathway and beta = 0.35 A(-1) for the triplet CR pathway. MFEs and TREPR experiments were used to measure the spin-spin exchange interaction, 2J, which is directly related to the electronic coupling matrix element for CR, V(CR)(2). The magnitude of 2J also shows an exponential distance dependence with a damping coefficient alpha = 0.36 A(-1), which agrees with the beta values obtained from the distance dependence for triplet CR. These results were analyzed in terms of the bridge molecular orbitals that participate in the charge transport mechanism. PMID- 19904985 TI - Use of methanol for the efficient extraction and analysis of melamine and cyanuric acid residues in dairy products and pet foods. AB - The recent worldwide shortage of acetonitrile has prompted the development of a new method using methanol as an alternative organic solvent in the extraction and liquid chromatographic analysis of melamine and cyanuric acid that may be present as contaminants in dairy products and pet foods. A simple extraction of melamine and cyanuric acid residues in fortified samples was successfully achieved, using a methanol-water mixture and analysis by isotopic dilution high-performance liquid chromatography-triple-quadrupole mass spectrometry (HPLC-MS/MS). A two step centrifugation procedure was employed to remove matrix components from extracts. The separation of melamine and cyanuric acid was carried out on a Dionex Acclaim Trinity P1 column, with a methanol and ammonium acetate buffer used as the mobile phase. Excellent linearity was achieved for both the melamine and cyanuric acid calibrations. A variety of dairy products and pet foods were fortified with melamine and cyanuric acid at three levels, 1, 2.5, and 10 microg/g, producing recovery yields of 101-119% for melamine and 84-123% for cyanuric acid. The lower limit of quantification (LLOQ) of melamine was 0.03 microg/g for liquid milk and 0.05 microg/g for dry infant milk formula. The quantitative results were comparable with those derived from previous methods that have been proposed by the U.S. Food and Drug Administration for the screening of melamine and its analogues in foods. PMID- 19904986 TI - Analyzing the molecular weight distribution in supramolecular polymers. AB - We have investigated the formation process of supramolecular linear polymer chains and its influence on the resulting chain length distribution function. For this purpose, we explored the migration of excitation energy between oligofluorene units coupled together through quadruple hydrogen-bonding groups to form linear chains that are terminated by oligophenylene vinylene end-caps acting as energy traps. The energy transfer dynamics from the main chain to the chain end was monitored experimentally using time-resolved PL spectroscopy and compared to an equivalent Monte Carlo simulation incorporating information on the structure of the chains, the transition transfer rates, and various weight distribution trial functions. We find that the assumption of a Flory distribution of chain lengths leads to excellent agreement between experimental and simulated data for a wide range of end-cap concentrations. On the other hand, both a Poisson function and a simplified assumption of a monodisperse distribution significantly underestimate the presence of long chains in the ensemble. Our results therefore show that supramolecular polymerization is a steplike process equivalent to polycondensation reactions in linear covalent polymers. These findings emphasize that equal reactivity of the supramolecular building blocks leads to a dynamic growth process for the supramolecular chain involving all chain components at all times. PMID- 19904987 TI - Chromium speciation analysis in bread samples. AB - Chromium is a controversial element with important essentiality and toxicity, depending on its different species; its speciation analysis in principal human foodstuffs, as in the case of bread, is of utmost importance. With this purpose, a method was validated, including a wet acid digestion procedure for total chromium dissolution, a selective alkaline extraction of hexavalent chromium, and ETAAS determination. The method was applied to the determination of total and hexavalent chromium in 152 bread samples. The total chromium contents were 47.3 +/- 20.0 and 50.9 +/- 22.2 microg/kg of dry weight for white and whole bread samples, respectively; those for hexavalent chromium were 5.65 +/- 5.44 and 6.82 +/- 4.88 microg/kg of dry weight. On the basis of a mean daily ingestion of three bread units, the calculated daily intake was up to 12.7 microg/day for total chromium and 1.98 microg/day for hexavalent chromium. Referring to total chromium, bread can contribute up to 10% of the Reference Daily Intake, 120 microg/day. PMID- 19904988 TI - Original and residual phytotoxicity of olive mill wastewater revealed by fractionations before and after incubation with Pleurotus ostreatus. AB - Concentrations of dissolved organic carbon (DOC) and total phenols (TP), and the phytotoxicity to cress (Lepidium sativum L.) were determined for three molecular sized fractions of olive mill wastewater (OMW), <1000, 1000-5000, and >5000 Da, before and after incubation with Pleurotus ostreatus. The <1000-Da fraction contained 82% of the total DOC and 48% of the TP, and was the most phytotoxic. Ethyl acetate separation of aqueous and solvent fractions showed that the aqueous fraction contained 93% of the total DOC, 83% of the TP, and was most phytotoxic, indicating low importance of monomeric phenols. Incubation of whole OMW and of the separate size fractions with P. ostreatus mycelia reduced TP by factors of 4.3-5.3, but exerted diverse impact on phytotoxicity; overall, P. ostreatus efficacy in organic load removal and OMW detoxification was limited. Additional size fractionation of the incubated fractions revealed that most residual phytotoxicity was associated with low-molecular weight (MW) compounds originated from the <1000 Da fraction and not with low-MW byproducts from the degradation of higher-MW fractions and that polymerized metabolites were nonphytotoxic. Total phenols should not be used as sole indicators of the successful remediation of OMW. PMID- 19904989 TI - Aptamer-nanoparticle strip biosensor for sensitive detection of cancer cells. AB - We report an aptamer-nanoparticle strip biosensor (ANSB) for the rapid, specific, sensitive, and low-cost detection of circulating cancer cells. Known for their high specificity and affinity, aptamers were first selected from live cells by the cell-SELEX (systematic evolution of ligands by exponential enrichment) process. When next combined with the unique optical properties of gold nanoparticles (Au-NPs), ANSBs were prepared on a lateral flow device. Ramos cells were used as a model target cell to demonstrate proof of principle. Under optimal conditions, the ANSB was capable of detecting a minimum of 4000 Ramos cells without instrumentation (visual judgment) and 800 Ramos cells with a portable strip reader within 15 min. Importantly, ANSB has successfully detected Ramos cells in human blood, thus providing a rapid, sensitive, and low-cost quantitative tool for the detection of circulating cancer cells. ANSB therefore shows great promise for in-field and point-of-care cancer diagnosis and therapy. PMID- 19904990 TI - Petroleum crude oil characterization by IMS-MS and FTICR MS. AB - Here, complementary ion mobility/mass spectrometry (IM/MS) and ultrahigh resolution Fourier transform ion cyclotron resonance (FTICR) MS analyses of light, medium, and heavy petroleum crude oils yielded distributions of the heteroatom-containing hydrocarbons, as well as multiple conformational classes. The IM/MS technique provides unique fingerprints for fast identification of signature conformational/compositional patterns, whereas FTICR MS analysis provides comprehensive heteroatom class distributions. IM/MS and FTICR MS results reveal an increase in compositional complexity in proceeding from light to medium to heavy crude oils. Inspection of the mobility results shows a high structural diversity for the C(n)H(h)XY (XY = N(1), S(1), N(1), O(1), NS, SO(1-2), NO(1-2), etc.) series, as well as a shift from planar to more compact three-dimensional structures with increasing mass. PMID- 19904992 TI - Architecture and dynamics of C18 bonded interphases with small molecule spacers. AB - The relationship between alkyl phase structure and chromatographic performance is investigated for a series of octadecyl (C(18))-modified silica surfaces with defined spacing of the alkyl surface by a "pre-end-capping" technique. Stationary phases were prepared by a two step process with (1) reaction with less than stochiometric amounts of a small monofunctional silane, followed by (2) solution or surface polymerization with octadecyltrichlorosilane. The results of solid state and suspension nuclear magnetic resonance (NMR) spectroscopy are correlated with the chromatographic behavior regarding shape selective separations. Two sets of six different stationary phases were prepared by solution and surface polymerization approaches, yielding materials with surface coverages from 2.7 to 5.6 micromol/m(2). (13)C cross-polarization magic angle spinning (CP/MAS) NMR spectra show a predominance of trans conformations for the set of surface polymerized phases with a C(18) coverage greater than 4.5 micromol/m(2). For the solution polymerized phases, no predominance for the trans conformation was observed, even for surface coverages greater than 5.1 micromol/m(2). Proton spectra in suspension indicate the trend that a higher coverage for the surface polymerized materials correlates with a more rigid alkyl chain conformation. The set of solution polymerized stationary phases confirms this tendency but minor deviations are observed for high coverages. These structural abnormalities are confirmed by differences in the (29)Si CP/MAS spectra. Furthermore, the (29)Si CP/MAS spectra indicate a lower amount of cross-linking for the materials with the highest amount of placeholder (spacer). The use of the different spectroscopic and chromatographic methods provides a wealth of information on the surface morphology of the systematically prepared C(18) materials and extends the understanding of surface morphology of alkyl modified silica and its influences of the molecular recognition process in liquid chromatography. PMID- 19904991 TI - Asymmetric construction of rings A-D of daphnicyclidin-type alkaloids. AB - The aza-Cope-Mannich reaction and ring-closing metathesis are key steps in the assembly of intermediates containing rings A-D of Daphniphyllum alkaloids of the daphnicyclidin type such as daphnipaxinin and oldhamine A. PMID- 19904993 TI - Patternable nanowire sensors for electrochemical recording of dopamine. AB - Spatially resolved electrochemical recording of neurochemicals is difficult due to the challenges associated with producing nanometer-scale patternable and integrated sensors. We describe the lithographic fabrication and characterization of patternable gold (Au) nanowire (NW) based sensors for the electrochemical recording of dopamine (DA). We demonstrate a straightforward NW-size-independent approach to align contact pads to NWs. Sensors, with NW widths as small as 30 nm, exhibited considerable insensitivity to scan rates during cyclic voltammetry, a nonlinear increase in oxidation current with increasing NW width, and the selectivity to measure submaximal synaptic concentrations of DA in the presence of interfering ascorbic acid. The electrochemical sensitivity of Au NW electrode sensors was much larger than that of Au thin-film electrodes. In chronoamperometric measurements, the NW sensors were found to be sensitive for submicromolar concentration of DA. Hence, the patternable NW sensors represent an attractive platform for electrochemical sensing and recording. PMID- 19904995 TI - Methicillin-resistant Staphylococcus aureus (MRSA)-active metabolites from Platanus occidentalis (American Sycamore). AB - One known and three new potent, selective, and nontoxic anti-MRSA metabolites, kaempferol 3-O-alpha-l-(2'',3''-di-E-p-coumaroyl)rhamnoside (1) (IC(50) 2.0 microg/mL), kaempferol 3-O-alpha-l-(2''-E-p-coumaroyl-3''-Z-p coumaroyl)rhamnoside (2) (IC(50) 0.8 microg/mL), kaempferol 3-O-alpha-l-(2''-Z-p coumaroyl-3''-E-p-coumaroyl)rhamnoside (3) (IC(50) 0.7 microg/mL), and kaempferol 3-O-alpha-l-(2'',3''-di-Z-p-coumaroyl)rhamnoside (4) (IC(50) 0.4 microg/mL), were isolated from the leaves of the common American sycamore, Platanus occidentalis. Compounds 2-4 are new. Due to the unusual selectivity, potency, and safety of the pure compounds and the semipure glycoside mixture against MRSA, it is clear that this represents a viable class of inhibitors to prevent growth of MRSA on surfaces and systemically. PMID- 19904994 TI - Rapid LC-MS drug metabolite profiling using microsomal enzyme bioreactors in a parallel processing format. AB - Silica nanoparticle bioreactors featuring thin films of enzymes and polyions were utilized in a novel high-throughput 96-well plate format for drug metabolism profiling. The utility of the approach was illustrated by investigating the metabolism of the drugs diclofenac (DCF), troglitazone (TGZ), and raloxifene, for which we observed known metabolic oxidation and bioconjugation pathways and turnover rates. A broad range of enzymes was included by utilizing human liver (HLM), rat liver (RLM) and bicistronic human-cyt P450 3A4 (bicis.-3A4) microsomes as enzyme sources. This parallel approach significantly shortens sample preparation steps compared to an earlier manual processing with nanoparticle bioreactors, allowing a range of significant enzyme reactions to be processed simultaneously. Enzyme turnover rates using the microsomal bioreactors were 2-3 fold larger compared to using conventional microsomal dispersions, most likely because of better accessibility of the enzymes. Ketoconazole (KET) and quinidine (QIN), substrates specific to cyt P450 3A enzymes, were used to demonstrate applicability to establish potentially toxic drug-drug interactions involving enzyme inhibition and acceleration. PMID- 19904996 TI - Lamiridosins, hepatitis C virus entry inhibitors from Lamium album. AB - Phytochemical study of the aqueous extract of the flowering tops of Lamium album led to identification of the antiviral iridoid isomers lamiridosins A and B (1, 2). These compounds were found to significantly inhibit hepatitis C virus entry (IC(50) 2.31 muM) in vitro. Studies of 14 iridoid analogues showed that, while the parent iridoid glucosides demonstrated no anti-HCV entry activity, the aglycones of shanzhiside methyl ester (4), loganin (5), loganic acid (6), geniposide (10), verbenalin (12), eurostoside (15), and picroside II (17) exhibited significant anti-HCV entry and anti-infectivity activities. PMID- 19904997 TI - Set-reset flip-flop memory based on enzyme reactions: toward memory systems controlled by biochemical pathways. AB - The enzyme-based set-reset flip-flop memory system was designed with the core part composed of horseradish peroxidase and diaphorase biocatalyzing oxidation and reduction of redox species (2,6-dichloroindophenol or ferrocyanide). The biocatalytic redox reactions were activated by H(2)O(2) and NADH produced in situ by different enzymatic reactions allowing transformation of various biochemical signals (glucose, lactate, d-glucose-6-phosphate, ethanol) into reduced or oxidized states of the redox species. The current redox state of the system, controlled by the set and reset signals, was read out by optical and electrochemical means. The multiwell setup with the flip-flop units separately activated by various set/reset signals allowed encoding of complex information. For illustrative purposes, the words "Clarkson" and then "University" were encoded using ASCII character codes. The present flip-flop system will allow additional functions of enzyme-based biocomputing systems, thus enhancing the performance of multisignal biosensors and actuators controlled by logically processed biochemical signals. The integrated enzyme logic systems and flip-flop memories associated with signal-responsive chemical actuators are envisaged as basic elements of future implantable biomedical devices controlled by immediate physiological conditions. PMID- 19904998 TI - Two-terminal nonvolatile memories based on single-walled carbon nanotubes. AB - Reproducible current hysteresis is observed in semiconducting single-walled carbon nanotubes (SWCNTs) measured in a two-terminal configuration without a gate electrode. On the basis of this hysteresis, a two-terminal nonvolatile memory is realized by applying voltage pulses of opposite polarities across the SWCNT. Charge trapping at the SWCNT/SiO(2) interface is proposed to account for the observed phenomena; this explanation is supported by the direct correlation between the switching behaviors and SWCNT carrier types. In particular, a change in dominant carrier type induced by adsorbates in air leads to the direct transition of hysteresis evolution in the same device, providing further evidence for the proposed mechanism. PMID- 19904999 TI - Interfacing microchip electrophoresis to a growth tube particle collector for semicontinuous monitoring of aerosol composition. AB - Semicontinuous monitoring of aerosol chemical composition has continually increased in demand because of the high spatial and temporal variability of atmospheric particles and the effects these aerosols have on human health and the environment. To address this demand, we describe the preliminary development of a semicontinuous aerosol composition analyzer consisting of a growth tube particle collector coupled to a microfluidic device for chemical analysis. The growth tube enlarges particles through water condensation in a laminar flow, permitting inertial collection into the microchip sample reservoir. Analysis is done by electrophoresis with conductivity detection. To avoid hydrodynamic interference from the sampling pressure, the microchip was operated isobarically by sealing the buffer reservoirs from the atmosphere and interconnecting all the reservoirs with air ducts. The collector samples at 1 L min(-1) and deposits particles into 30 microL of solution. Sample accumulates with time, and sequential injections are performed as aerosol concentration increases. For extended analyses, a sample rinsing system flushes the sample collection reservoir periodically. For inorganic anions, temporal resolution of 1 min and estimated detection limits of 70-140 ng m(-3) min were obtained. The system was used to measure sulfate and nitrate, and results were compared to a particle-into-liquid-sampler running in parallel. Results indicate that the prototype growth tube-microchip system (termed aerosol chip electrophoresis, ACE) could provide a useful complement to existing aerosol monitoring technologies, especially when less expensive and/or rapid analyses are desired. PMID- 19905000 TI - Visible light water splitting using dye-sensitized oxide semiconductors. AB - Researchers are intensively investigating photochemical water splitting as a means of converting solar to chemical energy in the form of fuels. Hydrogen is a key solar fuel because it can be used directly in combustion engines or fuel cells, or combined catalytically with CO(2) to make carbon containing fuels. Different approaches to solar water splitting include semiconductor particles as photocatalysts and photoelectrodes, molecular donor-acceptor systems linked to catalysts for hydrogen and oxygen evolution, and photovoltaic cells coupled directly or indirectly to electrocatalysts. Despite several decades of research, solar hydrogen generation is efficient only in systems that use expensive photovoltaic cells to power water electrolysis. Direct photocatalytic water splitting is a challenging problem because the reaction is thermodynamically uphill. Light absorption results in the formation of energetic charge-separated states in both molecular donor-acceptor systems and semiconductor particles. Unfortunately, energetically favorable charge recombination reactions tend to be much faster than the slow multielectron processes of water oxidation and reduction. Consequently, visible light water splitting has only recently been achieved in semiconductor-based photocatalytic systems and remains an inefficient process. This Account describes our approach to two problems in solar water splitting: the organization of molecules into assemblies that promote long-lived charge separation, and catalysis of the electrolysis reactions, in particular the four-electron oxidation of water. The building blocks of our artificial photosynthetic systems are wide band gap semiconductor particles, photosensitizer and electron relay molecules, and nanoparticle catalysts. We intercalate layered metal oxide semiconductors with metal nanoparticles. These intercalation compounds, when sensitized with [Ru(bpy)(3)](2+) derivatives, catalyze the photoproduction of hydrogen from sacrificial electron donors (EDTA(2-)) or non sacrificial donors (I(-)). Through exfoliation of layered metal oxide semiconductors, we construct multilayer electron donor-acceptor thin films or sensitized colloids in which individual nanosheets mediate light-driven electron transfer reactions. When sensitizer molecules are "wired" to IrO(2).nH(2)O nanoparticles, a dye-sensitized TiO(2) electrode becomes the photoanode of a water-splitting photoelectrochemical cell. Although this system is an interesting proof-of-concept, the performance of these cells is still poor (approximately 1% quantum yield) and the dye photodegrades rapidly. We can understand the quantum efficiency and degradation in terms of competing kinetic pathways for water oxidation, back electron transfer, and decomposition of the oxidized dye molecules. Laser flash photolysis experiments allow us to measure these competing rates and, in principle, to improve the performance of the cell by changing the architecture of the electron transfer chain. PMID- 19905001 TI - Simple and convenient approach to the Kreohnke pyridine type synthesis of functionalized indol-3-yl pyridine derivatives using 3-cyanoacetyl indole. AB - Access to privileged heterocyclic scaffolds involving 4-aryl-6-(1H-indol-3-yl) 2,2-bipyridine-5-carbonitriles and 6-(2-furyl)-2-(1H-indol-3-yl)-4-arylpyridine-3 carbonitriles frameworks has been achieved via a single-step multicomponent reaction of structurally diverse aldehydes, 2-acetylpyridine (or) 2-acetylfuran and 3-cyanoacetyl indole in ammonium acetate under neat condition. Also a series of 6,6'-di(1H-indol-3-yl)-4,4'-diaryl-2,2'-bipyridine-5,5'-dicarbonitrile and 7,7,7',7'-tetramethyl-4,4'-bis(aryl)-4,6,7,8,4',6',7',8'-octahydro-1H,1H [2,2']biquinolinyl-5,5'-dione derivatives are synthesized using cinnamil, 3 cyanoacetyl indole (or) dimedone and ammonium acetate. PMID- 19905002 TI - Bromo-directed N-2 alkylation of NH-1,2,3-triazoles: efficient synthesis of poly substituted 1,2,3-triazoles. AB - Reaction of 4-bromo-NH-1,2,3-triazoles 2 with alkyl halides in the presence of K(2)CO(3) in DMF produced the corresponding 2-substituted 4-bromo-1,2,3-triazoles 5 in a regioselective process. Subsequent Suzuki cross-coupling reaction of these bromides provided an efficient synthesis of 2,4,5-trisubstituted triazoles 3. In addition, reduction of the bromotriazoles by hydrogenation furnished an efficient synthesis of 2,4-disubstituted triazoles 8. PMID- 19905003 TI - Modular construction of 2-substituted benzo[b]furans from 1,2-dichlorovinyl ethers. AB - (E)-1,2-Dichlorovinyl ethers and amides are easily accessible from trichloroethylene via nucleophilic addition across in situ synthesized dichloroacetylene. A one-pot, sequential Suzuki-Miyaura coupling/intramolecular direct arylation between dichlorovinyl ethers and organoboronic acids provides easy access to a variety of benzofurans in only two steps from inexpensive commercially available compounds. The method is extendable to the preparation of indoles from the analogous dichlorovinyl amides. PMID- 19905004 TI - Pd(II)/(t)Bu-quinolineoxazoline: an air-stable and modular chiral catalyst system for enantioselective oxidative cascade cyclization. AB - An air-stable and structurally tunable chiral (t)Bu-quinolineoxazoline/Pd(II) catalyst system has been developed for the enantioselective oxidative cascade cyclization of a variety of disubstituted olefinic substrates, with the apparent advantages of good yields and excellent enantioselectivities (up to 98% ee) and diastereoselectivities (dr >24:1). A transition-state model has also been proposed to account for the excellent stereocontrol. PMID- 19905005 TI - Ring-size-selective enyne metathesis as a tool for desymmetrization of an enantiopure C2-symmetric building block. AB - The enantiomerically pure C(2)-symmetrical hexa-1,5-diene-3,4-diol is selectively monopropargylated. The products undergo ring-closing enyne metathesis to give exclusively dihydropyrans as single stereoisomers. An unprotected hydroxy group is identified as the factor controlling the ring-size selectivity. PMID- 19905006 TI - Site-specific installation and study of electroactive units in every layer of dendrons. AB - Whereas encapsulation of functional groups at the core of dendrimers is well understood, very little is known about their intermediate layers or even the periphery. Here we report on a systematic investigation of every layer of dendrimers by incorporating a single ferrocene unit in well-defined locations in dendrons. Site-specific incorporation of the ferrocene unit was achieved by utilizing the dendrimer sequencing methodology. We show here that the redox potential values of ferrocene at intermediate layers were remarkably different from those at the core and the periphery. Although redox potential values were location-dependent, no significant change in the rate of heterogeneous electron transfer (k(0)) was observed with respect to locations. This was attributed to the possibility that free rotation of dendrimer nullifies the distance between the electrode and ferrocene unit. Finally, we also show that no Faradaic current was observed for the amphiphilic assemblies of these dendrons, whereas the same dendron did exhibit significant Faradaic current in nonassembling solvent environments. PMID- 19905007 TI - Enhanced activity of enzymes immobilized in thermoresponsive core-shell microgels. AB - We present a quantitative study of the catalytic activity of beta-d-glucosidase from almonds adsorbed on thermosensitive microgels. The core-shell particles used as a carrier system consist of a solid polystyrene core onto which a poly(N isopropylacrylamide) (PNiPA) network is grafted. In the swollen state of this microgel, i.e., below the critical solution temperature (LCST) of PNiPA, high amounts of enzyme can be immobilized into the PNiPA network without loss of colloidal stability. The enzymatic activity of beta-d-glucosidase in its native form and in the adsorbed state was analyzed in terms of Michaelis-Menten kinetics. Moreover, the dependence of the enzymatic activity on temperature was investigated. We demonstrate that the enzymatic activity of beta-d-glucosidase adsorbed on such a core-shell microgel is increased by a factor of more than three compared to its activity in solution. This is in marked contrast to other carrier systems that usually lead to a strong decrease of enzymatic activity. Both the high loading capacity of the carrier observed and the increase of the catalytic activity of immobilized beta-d-glucosidase are traced back to the formation of strong interactions between the enzyme and microgel. Studies by Fourier-transform infrared (FT-IR) spectroscopy identify the formation of hydrogen bonds as driving forces for the adsorption. Hydrogen bonding may also be the reason for the enhanced activity. PMID- 19905008 TI - The involvement of intracellular calcium in the MCT-mediated uptake of lactic acid by HeLa cells. AB - The main object of this study was to evaluate the role of intracellular free calcium ion [Ca2+](in) in monocarboxylate transporter (MCT) mediated drug uptake by HeLa cells. It was hypothesized that alterations in the [Ca2+](in) levels affect Na+-H+ exchanger (NHE) regulated pH(in) and thereby produce the proton motivated driving force for monocarboxylate mediated substrate transport. The changes in intracellular pH (pH(in)) and MCT mediated uptake rates of L-lactic acid by HeLa cells, a human cervical adenocarcinoma cell line, were evaluated under the conditions, whose [Ca2+](in) concentrations were altered by various calcium modulators, such as EGTA-AM (a chelator), nifedipine (a Ca2+ channel antagonist) and A23187 (an ionophore). For the purpose of comparison, the L lactic acid uptake by HeLa cells was also evaluated under various pH(in) conditions induced by dexamethasone. The effects of the extracellular sodium concentration on the L-lactic acid uptake by HeLa cells were evaluated to determine the involvement of NHE-regulated pH changes in the MCT mediated drug uptake process. The [Ca2+](in) concentrations and pH(in) in HeLa were assessed using fluorescent probes fura-2 and 2',7'-bis[2-carboxyethyl-5 carboxyfluorescein] (BCECF), respectively. The treatment of HeLa cells with A23187 at concentrations of 50 and 100 microM enhanced [Ca2+](in) by 100% and 200% of the control, respectively. EGTA/AM (50 microM) or nifedipine (100 microM) did not cause any significant changes in the [Ca2+](in) levels, whereas EGTA/AM (100 microM) and nifedipine (200 microM) reduced the [Ca2+](in) levels by 30% and 25%, respectively, as compared with the control. A23187 at a concentration of 100 microM in the incubation medium lowered pH(in) (pH 5) and subsequently the uptake rate of lactic acid by 50% (0.47 +/- 0.03 micromol/mg protein/min) of the control. In contrast, nifedipine (200 microM) and EGTA-AM (100 microM), the calcium modulators that lowered the [Ca2+](in) levels and maintained the higher pH(in) (pH > 6) of HeLa cells, enhanced the uptake rate of lactic acid by 60% and 130% of the control, respectively. The results of this study demonstrated that there was a close correlation between the [Ca2+](in) level and pH(in) and that NHEs were involved with the MCT mediated uptake process in HeLa cells. An understanding of the role of [Ca2+](in) in the MCT mediated transport process could provide an efficient strategy to improve the systemic delivery of monocarboxylate substrates through the cervical mucosa. PMID- 19905009 TI - Microsecond time-resolved circular dichroism of rhodopsin photointermediates. AB - Time-resolved circular dichroism measurements, over a spectral range from 300 to 700 nm, were made at delays of 5, 100, and 500 micros after room-temperature photoexcitation of bovine rhodopsin in a lauryl maltoside suspension. The purpose was to provide more structural information about intermediate states in the activation of rhodopsin and other G protein-coupled receptors. In particular, information was sought about photointermediates that are isochromic or nearly isochromic in their unpolarized absorbance. The circular dichroism spectrum of lumirhodopsin, obtained after correcting the 5 micros difference CD data for the bleached rhodopsin, was in reasonable agreement with the lumirhodopsin CD spectrum obtained previously by thermal trapping at -76 degrees C. Similarly, the metarhodopsin II spectrum obtained with a 500 micros delay was also in agreement with the results of previous work on the temperature-trapped form of metarhodopsin II. However, the CD of the mixture formed with a 100 micros delay after photoexcitation, whose only visible absorbing component is lumirhodopsin, could not be accounted for near 480 nm in terms of the initially formed, 5 micros lumirhodopsin CD spectrum. Thus, the CD spectrum of lumirhodopsin changes on the time scale from 5 to 100 micros, showing reduced rotational strength in its visible band, possibly associated with either a process responsible for a small spectral shift that occurs in the lumirhodopsin absorbance spectrum at earlier times or the Schiff base deprotonation-reprotonation which occurs during equilibration of lumirhodopsin with the Meta I(380) photointermediate. Either explanation suggests a chromophore conformation change closely associated with deprotonation which could be the earliest direct trigger of activation. PMID- 19905011 TI - Helical polymers: synthesis, structures, and functions. PMID- 19905013 TI - Optimizing conical intersections by spin-flip density functional theory: application to ethylene. AB - Conical intersections (CIs) of ethylene have been successfully determined using spin-flip density functional theory (SFDFT) combined with a penalty-constrained optimization method. We present in detail three structures, twisted pyramidalized, hydrogen-migrated, and ethylidene CIs. In contrast to the linear response time-dependent density functional theory, which predicts a purely twisted geometry without pyramidalization as the S(1) global minimum, SFDFT gives a pyramidalized structure. Therefore, this is the first correct optimization of CI points of twisted ethylene by the DFT method. The calculated energies and geometries are in good agreement with those obtained by the multireference configuration interaction (MR-CI) method and the multistate formulation of second order multireference perturbation theory (MS-CASPT2). PMID- 19905014 TI - Vibrational dynamics of LiBH(4) by infrared pump-probe and 2D spectroscopy. AB - Employing femtosecond IR pump-probe and 2D spectroscopy, we measure the vibrational dynamics of LiBH(4) and several of its deuterium isotopomers. We find that the vibrational lifetime of various BH and BD stretching modes uniformly is approximately 1.5 ps for all BH(4-x)D(x)(-) units (0 2) it can strongly depend on the target position, and such dependence is explicitly calculated. PMID- 19905048 TI - Crossover transition in flowing granular chains. AB - We report on the dynamical and statistical behavior of flowing collections of granular chains confined two-dimensionally (2D) within a rotating tumbler. Experiments are conducted with systems of chains of fixed length, but various lengths are considered. The dynamics are punctuated by cascades of chains along a free-surface cascades, which drive the development of mixed porous/laminar packing arrangements in bulk. We investigate the conformation of the system, as characterized by the porosity of the flow region occupied by the chains and the mean-square end-to-end distance of the chains during flow. Both of these measures show crossover transitions from a 2D self-avoiding walk to a 2D random walk when the chain length becomes long enough to allow self-contact. PMID- 19905049 TI - Scenario for equilibrium solid-stabilized emulsions. AB - We show theoretically that under certain conditions colloidal particles can give rise to spontaneous emulsification of oil/water systems. The capillary penalty to create a large interface is compensated by entropic contributions connected to ionic dissociation on the colloid surfaces. The colloids themselves are absorbed on the oil/water interface. The conditions for spontaneous emulsification are: (1) oil-water interfacial tension is low (a few mN/m or lower); (2) interfacial tension between colloids and oil is smaller than between colloids and water (in the absence of charge effects); (3) density of chargeable groups on the colloids is large (order 1 nm-2); (4) Debye length is comparable to colloid size. PMID- 19905050 TI - Structure studies of the nematic phase formed by bent-core molecules. AB - In recent years there are several reports showing that bent-core mesogenic molecules are able to form biaxial nematic phase in which molecular rotation around the long molecular axis is strongly hindered. The x-ray pattern with azimuthally split signals at low angle region of diffraction is usually given as evidence for the biaxial nematic phase. We show experimentally and theoretically that such x-ray pattern is due to the local smectic- C fluctuations ("cybotactic" groups) in the uniaxial nematic phase. PMID- 19905051 TI - Deformation mechanism of nanocomposite gels studied by contrast variation small angle neutron scattering. AB - Contrast-variation small-angle neutron scattering (CV-SANS) was applied to investigate the deformation mechanism of high-performance nanocomposite polymer hydrogels (NC gels) consisting of polymer chains and inorganic clay platelets. Anisotropic SANS functions were obtained at various stretching ratios, lambda 's up to lambda=9 and were decomposed to three partial structure factors, S(ij)(Q parallel,Q perpendicular). Here, the subscripts i and j denote the polymer (P) or clay (C) and Q parallel and Q perpendicular are the magnitude of the scattering vectors along and perpendicular to the stretching directions, respectively. SCC(Q parallel,Q perpendicular) and S_{PP}(Q parallel,Q perpendicular) suggested that the orientation of clay platelets saturated by lambda approximately 3 , while the polymer chain stretching continued by further stretching. On the other hand, SCP(Q parallel,Q perpendicular) , only available by CV-SANS, indicated the presence of a polymer-enriched layer adsorbed to clay surface, which are responsible for large extensibility of NC gels over 1000% strain and large toughness exceeding 780 kPa. PMID- 19905052 TI - Lambda-prophage induction modeled as a cooperative failure mode of lytic repression. AB - We analyze a system-level model for lytic repression of lambda phage in E. coli using reliability theory, showing that the repressor circuit comprises four redundant components whose failure mode is prophage induction. Our model reflects the specific biochemical mechanisms involved in regulation, including long-range cooperative binding, and its detailed predictions for prophage induction in E. coli under ultraviolet radiation are in good agreement with experimental data. PMID- 19905053 TI - Robust ecological pattern formation induced by demographic noise. AB - We demonstrate that demographic noise can induce persistent spatial pattern formation and temporal oscillations in the Levin-Segel predator-prey model for plankton-herbivore population dynamics. Although the model exhibits a Turing instability in mean-field theory, demographic noise greatly enlarges the region of parameter space where pattern formation occurs. To distinguish between patterns generated by fluctuations and those present at the mean-field level in real ecosystems, we calculate the power spectrum in the noise-driven case and predict the presence of fat tails not present in the mean-field case. These results may account for the prevalence of large-scale ecological patterns, beyond that expected from traditional nonstochastic approaches. PMID- 19905054 TI - Cells, cancer, and rare events: homeostatic metastability in stochastic nonlinear dynamical models of skin cell proliferation. AB - A recently proposed model for skin cell proliferation [E. Clayton, Nature (London) 446, 185 (2007)] is extended to incorporate mitotic autoregulation, and hence homeostasis as a fixed point of the dynamics. Unlimited cell proliferation in such a model can be viewed as a model for carcinogenesis. One way in which this can arise is homeostatic metastability, in which the cell populations escape from the homeostatic basin of attraction by a large but rare stochastic fluctuation. Such an event can be viewed as the final step in a multistage model of carcinogenesis. Homeostatic metastability offers a possible explanation for the peculiar epidemiology of lung cancer in ex-smokers. PMID- 19905055 TI - Free-running period of neurons in the suprachiasmatic nucleus: Its dependence on the distribution of neuronal coupling strengths. AB - The suprachiasmatic nucleus (SCN) pacemaker shows a free-running period ranging from 20 to 28 h for different species, which was usually explained from the angle of coupling strength. Based on the assumption of nonidentical coupling strengths in SCN, we find an alternative mechanism that the diversity of free-running period can be also caused by the distribution of coupling strengths. The free running period is proportional to the average coupling strength and inverse proportional to the dispersion of couplings. Moreover, we present an analytic phase model to confirm our finding, which shows a solid foundation of our finding and opens a window to study the collective behaviors of SCN oscillators. PMID- 19905056 TI - Investigating quantum transport with an initial value representation of the semiclassical propagator. AB - Quantized systems whose underlying classical dynamics possess an elaborate mixture of regular and chaotic motion can exhibit rather subtle long-time quantum transport phenomena. In a short wavelength regime where semiclassical theories are most relevant, such transport phenomena, being quintessentially interference based, are difficult to understand with the system's specific long-time classical dynamics. Fortunately, semiclassical methods applied to wave packet propagation can provide a natural approach to understanding the connections, even though they are known to break down progressively as time increases. This is due to the fact that some long-time transport properties can be deduced from intermediate-time behavior. Thus, these methods need only retain validity and be carried out on much shorter time scales than the transport phenomena themselves in order to be valuable. The initial value representation of the semiclassical propagator of Herman and Kluk [Chem. Phys. 91, 27 (1984)] is heavily used in a number of molecular and atomic physics contexts, and is of interest here. It is known to be increasingly challenging to implement as the underlying classical chaos strengthens, and we ask whether it is possible to implement it well enough to extract the kind of intermediate-time information that reflects wave packet localization at long times. Using a system of two coupled quartic oscillators, we focus on the localizing effects of transport barriers formed by stable and unstable manifolds in the chaotic sea and show that these effects can be captured with the Herman-Kluk propagator. PMID- 19905057 TI - Coarse-graining the dynamics of a driven interface in the presence of mobile impurities: effective description via diffusion maps. AB - Developing effective descriptions of the microscopic dynamics of many physical phenomena can both dramatically enhance their computational exploration and lead to a more fundamental understanding of the underlying physics. Previously, an effective description of a driven interface in the presence of mobile impurities, based on an Ising variant model and a single empirical coarse variable, was partially successful [M. Haataja, Phys. Rev. Lett. 92, 160603 (2004)]; yet it underlined the necessity of selecting additional coarse variables in certain parameter regimes. In this paper we use a data mining approach to help identify the coarse variables required. We discuss the implementation of this diffusion map approach, the selection of a similarity measure between system snapshots required in the approach, and the correspondence between empirically selected and automatically detected coarse variables. We conclude by illustrating the use of the diffusion map variables in assisting the atomistic simulations and we discuss the translation of information between fine and coarse descriptions using lifting and restriction operators. PMID- 19905058 TI - Arbitrary-order corrections for finite-time drift and diffusion coefficients. AB - We address a standard class of diffusion processes with linear drift and quadratic diffusion coefficients. These contributions to dynamic equations can be directly drawn from data time series. However, real data are constrained to finite sampling rates and therefore it is crucial to establish a suitable mathematical description of the required finite-time corrections. Based on Ito Taylor expansions, we present the exact corrections to the finite-time drift and diffusion coefficients. These results allow to reconstruct the real hidden coefficients from the empirical estimates. We also derive higher-order finite time expressions for the third and fourth conditional moments that furnish extra theoretical checks for this class of diffusion models. The analytical predictions are compared with the numerical outcomes of representative artificial time series. PMID- 19905059 TI - Critical behavior of the Ising model on the four-dimensional cubic lattice. AB - In this paper we investigate the nature of the singularity of the Ising model of the four-dimensional cubic lattice. It is rigorously known that the specific heat has critical exponent alpha=0 but a nonrigorous field-theory argument predicts an unbounded specific heat with a logarithmic singularity at Tc. We find that within the given accuracy the canonical ensemble data are consistent both with a logarithmic singularity and a bounded specific heat but that the microcanonical ensemble lends stronger support to a bounded specific heat. Our conclusion is that either much larger system sizes are needed for Monte Carlo studies of this model in four dimensions or the field-theory prediction of a logarithmic singularity is wrong. PMID- 19905060 TI - Interactions leading to disordered ground states and unusual low-temperature behavior. AB - We have shown that any pair potential function v(r) possessing a Fourier transform V(k) that is positive and has compact support at some finite wave number K yields classical disordered ground states for a broad density range [R. D. Batten, F. H. Stillinger, and S. Torquato, J. Appl. Phys. 104, 033504 (2008)]. By tuning a constraint parameter chi (defined in the text), the ground states can traverse varying degrees of local order from fully disordered to crystalline ground states. Here, we show that in two dimensions, the " k -space overlap potential," where V(k) is proportional to the intersection area between two disks of diameter K whose centers are separated by k , yields anomalous low-temperature behavior, which we attribute to the topography of the underlying energy landscape. At T=0 , for the range of densities considered, we show that there is continuous energy degeneracy among Bravais-lattice configurations. The shear elastic constant of ground-state Bravais-lattice configurations vanishes. In the harmonic regime, a significant fraction of the normal modes for both amorphous and Bravais-lattice ground states have vanishing frequencies, indicating the lack of an internal restoring force. Using molecular-dynamics simulations, we observe negative thermal-expansion behavior at low temperatures, where upon heating at constant pressure, the system goes through a density maximum. For all temperatures, isothermal compression reduces the local structure of the system unlike typical single-component systems. PMID- 19905061 TI - Mapping of forced diffusion in quasi-one-dimensional systems. AB - Diffusion in an external potential in a two-dimensional channel of varying cross section is considered. We show that a rigorous mapping procedure applied on the corresponding Smoluchowski equation yields a one-dimensional evolution equation of the Fick-Jacobs type corrected by an effective coefficient D(x). The procedure enables us to derive this function within a recurrence scheme. We test this result on a model of stationary diffusion in a linear cone in a homogeneous potential, which is exactly solvable. Extension of the approximate formulas for D(x) derived for the diffusion alone is discussed. PMID- 19905062 TI - Explicit solution of the optimal fluctuation problem for an elastic string in a random medium. AB - The free-energy distribution function of an elastic string in a quenched random potential, PL(F) , is investigated with the help of the optimal fluctuation approach. The form of the far-right tail of PL(F) is found by constructing the exact solution of the nonlinear saddle-point equations describing the asymptotic form of the optimal fluctuation. The solution of the problem is obtained for two different types of boundary conditions and for an arbitrary dimension of the imbedding space 1+d with d from the interval 0>1 and k>>1 . For the J1>J2 case, the kinetic evolution of inert polymers is very complex and ck(t) can take one of the three forms: monotone decreasing, single peak (Poisson-like distribution), and double peak. For the special J1=J2 case, ck(t) exhibits an exponential decay in size. PMID- 19905070 TI - Phase transition in a long-range antiferromagnetic model. AB - We consider an Ising model where longitudinal components of every pair of spins have antiferromagnetic interaction of the same magnitude. When subjected to a transverse magnetic field at zero temperature, the system undergoes a phase transition of second order to an ordered phase and if the temperature is now increased, there is another phase transition to disordered phase. We provide derivation of these features by perturbative treatment up to the second order and argue that the results are nontrivial and not derivable from the known results about related models. PMID- 19905071 TI - Extended defects in the Potts-percolation model of a solid: renormalization group and Monte Carlo analysis. AB - We extend the model of a 2d solid to include a line of defects. Neighboring atoms on the defect line are connected by springs of different strength and different cohesive energy with respect to the rest of the system. Using the Migdal-Kadanoff renormalization group we show that the elastic energy is an irrelevant field at the bulk critical point. For zero elastic energy this model reduces to the Potts model. By using Monte Carlo simulations of the three- and four-state Potts model on a square lattice with a line of defects, we confirm the renormalization-group prediction that for a defect interaction larger than the bulk interaction the order parameter of the defect line changes discontinuously while the defect energy varies continuously as a function of temperature at the bulk critical temperature. PMID- 19905072 TI - Stochastic analysis of dimerization systems. AB - The process of dimerization, in which two monomers bind to each other and form a dimer, is common in nature. This process can be modeled using rate equations, from which the average copy numbers of the reacting monomers and of the product dimers can then be obtained. However, the rate equations apply only when these copy numbers are large. In the limit of small copy numbers the system becomes dominated by fluctuations, which are not accounted for by the rate equations. In this limit one must use stochastic methods such as direct integration of the master equation or Monte Carlo simulations. These methods are computationally intensive and rarely succumb to analytical solutions. Here we use the recently introduced moment equations which provide a highly simplified stochastic treatment of the dimerization process. Using this approach, we obtain an analytical solution for the copy numbers and reaction rates both under steady state conditions and in the time-dependent case. We analyze three different dimerization processes: dimerization without dissociation, dimerization with dissociation, and heterodimer formation. To validate the results we compare them with the results obtained from the master equation in the stochastic limit and with those obtained from the rate equations in the deterministic limit. Potential applications of the results in different physical contexts are discussed. PMID- 19905073 TI - Theory of single-file multiparticle diffusion in narrow pores. AB - Single-file diffusion of multiple strongly interacting particles in a one dimensional pore is described within a general analytical framework. The theory accounts for nonequilibrium conditions, explicit particle-particle interactions, external potential acting on the particles and the fluctuations of the number of particles due to their exchange with external equilibrium reservoirs. It is shown that the problem can be reduced to a closed hierarchical set of partial differential equations of increasing dimensionality, which can be solved numerically. Our framework allows computing any macroscopic characteristic of multiparticle diffusion in the pore. It is shown that the pore occupancy probabilities and the current are rational functions of external concentrations in the steady state. The theory is tested on a simplified model of the narrow rigid pore inspired by the selectivity filter of biological ion channel. Perspectives and limitations of the theory are discussed. PMID- 19905074 TI - Finite-size effects in presence of gravity: the behavior of the susceptibility in 3He and 4He films near the liquid-vapor critical point. AB - We study critical-point finite-size effects on the behavior of susceptibility of a film placed in the Earth's gravitational field. The fluid-fluid and substrate fluid interactions are characterized by van der Waals type power-law tails, and the boundary conditions are consistent with bounding surfaces that strongly prefer the liquid phase of the system. Specific predictions are made with respect to the behavior of 3He and 4He films in the vicinity of their respective liquid gas critical points. We find that for all film thicknesses of current experimental interest the combination of van der Waals interactions and gravity leads to substantial deviations from the behavior predicted by models in which all interatomic forces are very short ranged and gravity is absent. In the case of a completely short-ranged system exact mean-field analytical expressions are derived, within the continuum approach, for the behavior of both the local and the total susceptibilities. PMID- 19905075 TI - Glassy properties of frustrated arrays of nonlinear devices. AB - We study linear arrays of different number of quartic oscillators shaped in the form of a ring when Gaussian noise (temperature) is added. Frustration is introduced through periodic boundary conditions and repulsive, directional interactions between neighboring oscillators. We show that these systems have similar dynamic properties than the arrays of fluxgates magnetometers. We find that there is a critical number of oscillators separating the regimes arising for systems with few and many oscillators and show that they reach an optimum ordering for a nonvanishing temperature. We also find that they have a relaxation process with an infinite mean life that is typical of glassy systems. PMID- 19905076 TI - Geometry of phase separation. AB - We study the domain geometry during spinodal decomposition of a 50:50 binary mixture in two dimensions. Extending arguments developed to treat nonconserved coarsening, we obtain approximate analytic results for the distribution of domain areas and perimeters during the dynamics. The main approximation is to regard the interfaces separating domains as moving independently. While this is true in the nonconserved case, it is not in the conserved one. Our results can therefore be considered as a "first-order" approximation for the distributions. In contrast to the celebrated Lifshitz-Slyozov-Wagner distribution of structures of the minority phase in the limit of very small concentration, the distribution of domain areas in the 50:50 case does not have a cutoff. Large structures (areas or perimeters) retain the morphology of a percolative or critical initial condition, for quenches from high temperatures or the critical point, respectively. The corresponding distributions are described by a cA-tau tail, where c and tau are exactly known. With increasing time, small structures tend to have a spherical shape with a smooth surface before evaporating by diffusion. In this regime, the number density of domains with area A scales as A1/2 , as in the Lifshitz-Slyozov Wagner theory. The threshold between the small and large regimes is determined by the characteristic area A approximately t2/3. Finally, we study the relation between perimeters and areas and the distribution of boundary lengths, finding results that are consistent with the ones summarized above. We test our predictions with Monte Carlo simulations of the two-dimensional Ising model. PMID- 19905077 TI - Dynamics of a microorganism moving by chemotaxis in its own secretion. AB - The Brownian dynamics of a single microorganism coupled by chemotaxis to a diffusing concentration field that is secreted by the microorganism itself is studied by computer simulations in spatial dimensions d=1,2,3 . Both cases of a chemoattractant and a chemorepellent are discussed. For a chemoattractant, we find a transient dynamical arrest until the microorganism diffuses for long times. For a chemorepellent, there is a transient ballistic motion in all dimensions and a long-time diffusion. These results are interpreted with the help of a theoretical analysis. PMID- 19905078 TI - Topology, symmetry, phase transitions, and noncollinear spin structures. AB - We use a topological approach to describe the frustration- and field-induced phase transitions exhibited by the infinite-range XY model on the AB2 chain, including noncollinear spin structures. For this purpose, we have computed the Morse number and the Euler characteristic, as well as other topological invariants, which are found to behave similarly as a function of the energy level in the context of Morse theory. In particular, we use a method based on an analogy with statistical mechanics to compute the Euler characteristic, which proves to be quite feasible. We also introduce topological energies which help us to clarify several properties of the transitions, both at zero and finite temperatures. In addition, we establish a nontrivial direct connection between the thermodynamics of the systems, which have been solved exactly under the saddle-point approach, and the topology of their configuration space. This connection allows us to identify the nondegeneracy condition under which the divergence of the density of Jacobian's critical points [jl(E)] at the critical energy of a topology-induced phase transition, proposed by Kastner and Schnetz [Phys. Rev. Lett. 100, 160601 (2008)] as a necessary criterion, is suppressed. Finally, our findings and those available in the literature suggest that the cusplike singularity exhibited both by the Euler characteristic and the topological contribution for the entropy at the critical energy, put together with the divergence of jl(E) , and emerge as necessary and sufficient conditions for the occurrence of the finite-temperature topology-induced phase transitions examined in this work. The general character of this proposal should be subject to a more rigorous scrutiny. PMID- 19905079 TI - Dynamic scaling functions and amplitude ratios of stochastic models with energy conservation above Tc. AB - Dynamical scaling functions above Tc for the characteristic frequencies and the dynamical correlation functions of the order parameter and the conserved density of model C are calculated in one loop order. By a proper exponentiation procedure these results can be extended in order to consider the changes in these functions using the fixed point values and exponents in two loop order. The dynamical amplitude ratio R of the characteristic frequencies is generalized to the critical region. Surprisingly the decay of the shape functions at large scaled frequency does not behave as expected from applying scaling arguments. The exponent upsilon of the decay does not change when going from the critical to the hydrodynamic region although the shape functions change. The value of upsilon for the order parameter is in agreement with its value in the critical region, whereas for the conserved density it is equal to 2, the value in the hydrodynamic region. PMID- 19905080 TI - Random walks on deterministic scale-free networks: exact results. AB - We study the random walk problem on a class of deterministic scale-free networks displaying a degree sequence for hubs scaling as a power law with an exponent gamma=log 3/log 2. We find exact results concerning different first-passage phenomena and, in particular, we calculate the probability of first return to the main hub. These results allow to derive the exact analytic expression for the mean time to first reach the main hub, whose leading behavior is given by tau approximately V1-1/gamma, where V denotes the size of the structure, and the mean is over a set of starting points distributed uniformly over all the other sites of the graph. Interestingly, the process turns out to be particularly efficient. We also discuss the thermodynamic limit of the structure and some local topological properties. PMID- 19905081 TI - Critical exponents from cluster coefficients. AB - For a large class of repulsive interaction models, the Mayer cluster integrals can be transformed into a tridiagonal real symmetric matrix R_{mn} , whose elements converge to two constants. This allows for an effective extrapolation of the equation of state for these models. Due to a nearby (nonphysical) singularity on the negative real z axis, standard methods (e.g., Pade approximants based on the cluster integrals expansion) fail to capture the behavior of these models near the ordering transition, and, in particular, do not detect the critical point. A recent work [E. Eisenberg and A. Baram, Proc. Natl. Acad. Sci. U.S.A. 104, 5755 (2007)] has shown that the critical exponents sigma and sigma;{'} , characterizing the singularity of the density as a function of the activity, can be exactly calculated if the decay of the R matrix elements to their asymptotic constant follows a 1/n;{2} law. Here we employ renormalization group (RG) arguments to extend this result and analyze cases for which the asymptotic approach of the R matrix elements toward their limiting value is of a more general form. The relevant asymptotic correction terms (in RG sense) are identified, and we then present a corrected exact formula for the critical exponents. We identify the limits of usage of the formula and demonstrate one physical model, which is beyond its range of validity. The formula is validated numerically and then applied to analyze a number of concrete physical models. PMID- 19905082 TI - Kramers-Moyal coefficients in the analysis and modeling of heart rate variability. AB - Modeling of recorded time series may be used as a method of analysis for heart rate variability studies. In particular, the extraction of the first two Kramers Moyal coefficients has been used in this context. Recently, the method was applied to a wide range of signal analysis: from financial data to physiological and biological time series. Modeling of the signal is important for the prediction and interpretation of the dynamics underlying the process. The method requires the determination of the Markov time. Obtaining the drift and diffusion term of the Kramers-Moyal expansion is crucial for the modeling of the original time series with the Langevin equation. Both Tabar [Comput. Sci. Eng. 8, 54 (2006)] and T. Kuusela [Phys. Rev. E 69, 031916 (2004)] suggested that these terms may be used to distinguish healthy subjects from those with heart failure. The research groups applied a somewhat different methodology and obtained substantially different ranges of the Markov time. We show that the two studies may be considered consistent with each other as Kuusela analyzed 24 h recordings while Tabar analyzed daytime and nighttime recordings, separately. However, both groups suggested using the Langevin equation for modeling of time series which requires the fluctuation force to be a Gaussian. We analyzed heart rate variability recordings for ten young male (age 26-4+3 y ) healthy subjects. 24 h recordings were analyzed and 6-h-long daytime and nighttime fragments were selected. Similar properties of the data were observed in all recordings but all the nighttime data and seven of the ten 24 h series exhibited higher-order, non negligible Kramers-Moyal coefficients. In such a case, the reconstruction of the time series using the Langevin equation is impossible. The non-negligible higher order coefficients are due to autocorrelation in the data. This effect may be interpreted as a result of a physiological phenomenon (especially occurring for nighttime data): respiratory sinus arrhythmia (RSA). We detrended the nighttime recordings for the healthy subjects and obtained an asymmetry in the dependence of the diffusion term on the rescaled heart rate. This asymmetry seems to be an effect of different time scales during the inspiration and the expiration phase of breathing. The asymmetry was significantly decreased in the diffusion term found for detrended nighttime recordings obtained from five hypertrophic cardiomyopathy (HCM) patients. We conclude that the effect of RSA is decreased in the heart rate variability of HCM patients-a result which may contribute to a better medical diagnosis by supplying a new quantitative measure of RSA. PMID- 19905083 TI - Rocking feedback-controlled ratchets. AB - We investigate the different regimes that emerge when a periodic driving force, the rocking force, acts on a collective feedback flashing ratchet. The interplay of the rocking and the feedback control gives a rich dynamics with different regimes presenting several unexpected features. In particular, we show that for both the one-particle ratchet and the collective version of the ratchet an appropriate rocking increases the flux. This mechanism gives the maximum flux that has been achieved in a ratchet device without an a priori bias. PMID- 19905084 TI - Long delay times in reaction rates increase intrinsic fluctuations. AB - In spatially distributed cellular systems, it is often convenient to represent complicated auxiliary pathways and spatial transport by time-delayed reaction rates. Furthermore, many of the reactants appear in low numbers necessitating a probabilistic description. The coupling of delayed rates with stochastic dynamics leads to a probability conservation equation characterizing a non-Markovian process. A systematic approximation is derived that incorporates the effect of delayed rates on the characterization of molecular noise valid in the limit of long delay time. By way of a simple example, we show that delayed reaction dynamics can only increase intrinsic fluctuations about the steady state. The method is general enough to accommodate nonlinear transition rates allowing characterization of fluctuations around a delay-induced limit cycle. PMID- 19905085 TI - Dissipative dynamics of a harmonic oscillator: a nonperturbative approach. AB - Starting from a microscopic theory, we derive a master equation for a harmonic oscillator coupled to a bath of noninteracting oscillators. We follow a nonperturbative approach, proposed earlier by us for the free Brownian particle. The diffusion constants are calculated analytically and the positivity of the master equation is shown to hold above a critical temperature. We compare the long time behavior of the average kinetic and potential energies with known thermodynamic results. In the limit of vanishing oscillator frequency of the system, we recover the results of the free Brownian particle. PMID- 19905086 TI - Nonequilibrium thermodynamics of driven amorphous materials. I. Internal degrees of freedom and volume deformation. AB - This is the first of three papers devoted to the nonequilibrium thermodynamics of amorphous materials. Our focus here is on the role of internal degrees of freedom in determining the dynamics of such systems. For illustrative purposes, we study a solid whose internal degrees of freedom are vacancies that govern irreversible volume changes. Using this model, we compare a thermodynamic theory based on the Clausius-Duhem inequality to a statistical analysis based directly on the law of increase of entropy. The statistical theory is used first to derive the Clausius Duhem inequality. We then use the theory to go beyond those results and obtain detailed equations of motion, including a rate factor that is enhanced by deformation-induced noisy fluctuations. The statistical analysis points to the need for understanding how both energy and entropy are shared by the vacancies and their environments. PMID- 19905087 TI - Nonequilibrium thermodynamics of driven amorphous materials. II. Effective temperature theory. AB - We develop a theory of the effective disorder temperature in glass-forming materials driven away from thermodynamic equilibrium by external forces. Our basic premise is that the slow configurational degrees of freedom of such materials are weakly coupled to the fast kinetic-vibrational degrees of freedom and therefore that these two subsystems can be described by different temperatures during deformation. We use results from the preceding paper on the nonequilibrium thermodynamics of systems with internal degrees of freedom to derive an equation of motion for the effective temperature and to learn how this temperature couples to the dynamics of the system as a whole. PMID- 19905088 TI - Nonequilibrium thermodynamics of driven amorphous materials. III. Shear transformation-zone plasticity. AB - We use the internal-variable, effective-temperature thermodynamics developed in two preceding papers to reformulate the shear-transformation-zone (STZ) theory of amorphous plasticity. As required by the preceding analysis, we make explicit approximations for the energy and entropy of the STZ internal degrees of freedom. We then show that the second law of thermodynamics constrains the STZ transition rates to have an Eyring form as a function of the effective temperature. Finally, we derive an equation of motion for the effective temperature for the case of STZ dynamics. PMID- 19905089 TI - Thermal breakage of a discrete one-dimensional string. AB - We study the thermal breakage of a discrete one-dimensional string, with open and fixed ends, in the heavily damped regime. Basing our analysis on the multidimensional Kramers escape theory, we are able to make analytical predictions on the mean breakage rate and on the breakage propensity with respect to the breakage location on the string. We then support our predictions with numerical simulations. PMID- 19905090 TI - Site-diluted Ising model in four dimensions. AB - In the literature, there are five distinct fragmented sets of analytic predictions for the scaling behavior at the phase transition in the random-site Ising model in four dimensions. Here, the scaling relations for logarithmic corrections are used to complete the scaling pictures for each set. A numerical approach is then used to confirm the leading scaling picture coming from these predictions and to discriminate between them at the level of logarithmic corrections. PMID- 19905091 TI - One-dimensional hard-point gas as a thermoelectric engine. AB - We demonstrate the possibility to build a thermoelectric engine using a one dimensional gas of molecules with unequal masses and hard-point interaction. Most importantly, we show that the efficiency of this engine is determined by a parameter YT which is different from the well known figure of merit ZT . Even though the efficiency of this particular model is low, our results shed light on the problem and open the possibility to build efficient thermoelectric engines. PMID- 19905092 TI - Geometric and projection effects in Kramers-Moyal analysis. AB - Kramers-Moyal coefficients provide a simple and easily visualized method with which to analyze nonlinear stochastic time series. One mechanism that can affect the estimation of the coefficients is geometric projection effects. For some biologically inspired examples, these effects are predicted and explored with a nonstochastic projection operator method and compared with direct numerical simulation of the systems' Langevin equations. General features and characteristics are identified, and the utility of the Kramers-Moyal method is discussed. Projections of a system are in general non-Markovian, but here the Kramers-Moyal method remains useful, and in any case the primary examples considered are found to be close to Markovian. PMID- 19905093 TI - Equilibrium properties of disordered spin models with two-scale interactions. AB - Methods for understanding classical disordered spin systems with interactions conforming to some idealized graphical structure are well developed. The equilibrium properties of the Sherrington-Kirkpatrick model, which has a densely connected structure, have become well understood. Many features generalize to sparse Erdos-Renyi graph structures above the percolation threshold and to Bethe lattices when appropriate boundary conditions apply. In this paper, we consider spin states subject to a combination of sparse strong interactions with weak dense interactions, which we term a composite model. The equilibrium properties are examined through the replica method, with exact analysis of the high temperature paramagnetic, spin-glass, and ferromagnetic phases by perturbative schemes. We present results of replica symmetric variational approximations, where perturbative approaches fail at lower temperature. Results demonstrate re entrant behaviors from spin glass to ferromagnetic phases as temperature is lowered, including transitions from replica symmetry broken to replica symmetric phases. The nature of high-temperature transitions is found to be sensitive to the connectivity profile in the sparse subgraph, with regular connectivity a discontinuous transition from the paramagnetic to ferromagnetic phases is apparent. PMID- 19905094 TI - Equilibrium long-ranged charge correlations at the interface between media coupled to the electromagnetic radiation. AB - We continue studying long-ranged quantum correlations of surface charge densities on the interface between two media of distinct dielectric functions which are in thermal equilibrium with the radiated electromagnetic field. Two regimes are considered: the nonretarded one with the speed of light c taken to be infinitely large and the retarded one with a finite value of c . The analysis is based on our results obtained by using fluctuational electrodynamics [L. Samaj and B. Jancovici, Phys. Rev. E 78, 051119 (2008)]. Using an integration method in the complex plane and the general analytical properties of dielectric functions in the frequency upper half plane, we derive explicit forms of prefactors to the long-range decay of the surface charge correlation functions for all possible media (conductor, dielectric, and vacuum) configurations. The main result is that the time-dependent quantum prefactor in the retarded regime takes its static classical form for any temperature. PMID- 19905095 TI - Soft random solids and their heterogeneous elasticity. AB - Spatial heterogeneity in the elastic properties of soft random solids is examined via vulcanization theory. The spatial heterogeneity in the structure of soft random solids is a result of the fluctuations locked-in at their synthesis, which also brings heterogeneity in their elastic properties. Vulcanization theory studies semimicroscopic models of random-solid-forming systems and applies replica field theory to deal with their quenched disorder and thermal fluctuations. The elastic deformations of soft random solids are argued to be described by the Goldstone sector of fluctuations contained in vulcanization theory, associated with a subtle form of spontaneous symmetry breaking that is associated with the liquid-to-random-solid transition. The resulting free energy of this Goldstone sector can be reinterpreted as arising from a phenomenological description of an elastic medium with quenched disorder. Through this comparison, we arrive at the statistics of the quenched disorder of the elasticity of soft random solids in terms of residual stress and Lame-coefficient fields. In particular, there are large residual stresses in the equilibrium reference state, and the disorder correlators involving the residual stress are found to be long ranged and governed by a universal parameter that also gives the mean shear modulus. PMID- 19905096 TI - Harmonic measure for critical Potts clusters. AB - We present a technique, which we call "etching," which we use to study the harmonic measure of Fortuin-Kasteleyn clusters in the Q-state Potts model for Q=1 4 . The harmonic measure is the probability distribution of random walkers diffusing onto the perimeter of a cluster. We use etching to study regions of clusters which are extremely unlikely to be hit by random walkers, having hitting probabilities down to 10-4600. We find good agreement between the theoretical predictions of Duplantier and our numerical results for the generalized dimension D(q) including regions of small and negative q . PMID- 19905097 TI - Competition between multiple totally asymmetric simple exclusion processes for a finite pool of resources. AB - Using Monte Carlo simulations and a domain-wall theory, we discuss the effect of coupling several totally asymmetric simple exclusion processes (TASEPs) to a finite reservoir of particles. This simple model mimics directed biological transport processes in the presence of finite resources such as protein synthesis limited by a finite pool of ribosomes. If all TASEPs have equal length, we find behavior which is analogous to a single TASEP coupled to a finite pool. For the more generic case of chains with different lengths, several unanticipated regimes emerge. A generalized domain-wall theory captures our findings in good agreement with simulation results. PMID- 19905098 TI - Stochastic Langevin equations: Markovian and non-Markovian dynamics. AB - Non-Markovian stochastic Langevin-like equations of motion are compared to their corresponding Markovian (local) approximations. The validity of the local approximation for these equations, when contrasted with the fully nonlocal ones, is analyzed in detail. The conditions for when the equation in a local form can be considered a good approximation are then explicitly specified. We study both the cases of additive and multiplicative noises, including system-dependent dissipation terms, according to the fluctuation-dissipation theorem. PMID- 19905099 TI - Reducing the heterogeneity of payoffs: an effective way to promote cooperation in the prisoner's dilemma game. AB - In this paper, the accumulated payoff of each agent is regulated so as to reduce the heterogeneity of the distribution of all such payoffs. It is found that there exists an optimal regulation strength at which cooperation in the prisoner's dilemma game is optimally promoted. If the heterogeneity is regulated to be either too weak or too strong, the promotive effect disappears and the evolution of cooperation may even be impaired if compared to the absence of the proposed regulatory mechanism. An explanation of the observed results is provided. In particular, it is found that cooperators on the spatial grid are not isolated but form compact clusters and that the distribution of these clusters is crucial for the promotion of cooperation. Our work provides insights into relations between the distribution of payoffs and the evolution of cooperative behavior in situations constituting a social dilemma. PMID- 19905100 TI - Finite bath fluctuation theorem. AB - We demonstrate that a finite bath fluctuation theorem of the Crooks type holds for systems that have been thermalized via weakly coupling them to a bath with energy independent finite specific heat. We show that this theorem reduces to the known canonical and microcanonical fluctuation theorems in the two respective limiting cases of infinite and vanishing specific heat of the bath. The result is elucidated by applying it to a two-dimensional hard disk colliding elastically with few other hard disks in a rectangular box with perfectly reflecting walls. PMID- 19905101 TI - Robustness of optimal intermittent search strategies in one, two, and three dimensions. AB - Search problems at various scales involve a searcher, be it a molecule before reaction or a foraging animal, which performs an intermittent motion. Here we analyze a generic model based on such type of intermittent motion, in which the searcher alternates phases of slow motion allowing detection and phases of fast motion without detection. We present full and systematic results for different modeling hypotheses of the detection mechanism in space in one, two, and three dimensions. Our study completes and extends the results of our recent letter [Loverdo, Nat. Phys. 4, 134 (2008)] and gives the necessary calculation details. In addition, another modeling of the detection case is presented. We show that the mean target detection time can be minimized as a function of the mean duration of each phase in one, two, and three dimensions. Importantly, this optimal strategy does not depend on the details of the modeling of the slow detection phase, which shows the robustness of our results. We believe that this systematic analysis can be used as a basis to study quantitatively various real search problems involving intermittent behaviors. PMID- 19905102 TI - Noise-induced escape from bifurcating attractors: Symplectic approach in the weak noise limit. AB - The effect of noise is studied in one-dimensional maps undergoing transcritical, tangent, and pitchfork bifurcations. The attractors of the noiseless map become metastable states in the presence of noise. In the weak-noise limit, a symplectic two-dimensional map is associated with the original one-dimensional map. The consequences of their noninvertibility on the phase-space structures are discussed. Heteroclinic orbits are identified which play a key role in the determination of the escape rates from the metastable states. Near bifurcations, the critical slowing down justifies the use of a continuous-time approximation replacing maps by flows, which allows the analytic calculation of the escape rates. This method provides the universal scaling behavior of the escape rates at the bifurcations. PMID- 19905103 TI - Continuous-time multidimensional Markovian description of Levy walks. AB - The paper presents a multidimensional model for nonlinear Markovian random walks that generalizes the one we developed previously [I. Lubashevsky, R. Friedrich, and A. Heuer, Phys. Rev. E 79, 011110 (2009)] in order to describe the Levy-type stochastic processes in terms of continuous trajectories of walker motion. This approach may open a way to treat Levy flights or Levy random walks in inhomogeneous media or systems with boundaries in the future. The proposed model assumes the velocity of a wandering particle to be affected by a linear friction and a nonlinear Langevin force whose intensity is proportional to the magnitude of the velocity for its large values. Based on the singular perturbation technique, the corresponding Fokker-Planck equation is analyzed and the relationship between the system parameters and the Levy exponent is found. Following actually the previous paper we demonstrate also that anomalously long displacements of the wandering particle are caused by extremely large fluctuations in the particle velocity whose duration is determined by the system parameters rather than the duration of the observation interval. In this way we overcome the problem of ascribing to Levy random-walk non-Markov properties. PMID- 19905104 TI - Measurement of granular entropy. AB - Recently, Dean and Lefevre [Phys. Rev. Lett. 90, 198301 (2003)] developed a method for testing the statistical mechanical theory of granular packings proposed by Edwards and co-workers [Physica A 157, 1080 (1989); Phys. Rev. E 58, 4758 (1998)]. The method relies on the prediction that the ratio of two overlapping volume histograms should be exponential in volume. We extend the method by showing that one can also calculate the entropy of the packing and also that the method can yield false positive results when the histograms are Gaussians with nearly identical variances. We then apply the method to simulations and experiments of granular compaction. The distribution of global volumes (the volume of the entire packing) is nearly Gaussian and it is difficult to conclude if the theory is valid. On the other hand, the distribution of Voronoi volumes clearly obeys the theoretical prediction. PMID- 19905105 TI - Unsteady granular flows in a rotating tumbler. AB - The characteristics of steady granular flow in quasi-two-dimensional rotating tumblers have been thoroughly investigated and are fairly well understood. However, unsteady time-varying flow has not been studied in detail. The linear response of granular flow in quasi-two-dimensional rotating tumblers is presented for periodic forcing protocols via sinusoidal variation in the rotational speed of the tumbler and for step changes in rotational speed. Variations in the tumbler radius, particle size, and forcing frequency are explored. Similarities to steady flow include the fastest flow occurring at the free surface of the flowing layer and an instantaneous approximately linear velocity profile through the depth. The flowing layer depth varies by 2-5 particle diameters between minimum and maximum rotation rates. However, unsteady forcing also causes the flow to exhibit dynamic properties. For periodic rotational speeds, the phase lag of the flowing layer depth increases linearly with increasing input forcing frequency up to nearly 2.0 rad over 0-20 cycles per tumbler revolution. The amplitude responses of the velocity and shear rate show a resonance behavior unique to the system level parameters. The phase lag of all flow properties appears to be related to the number of particle contacts from the edge of the rotating tumbler. Characterization via step changes in rotational speed shows dynamic properties of overshoot (up to 35%) and rise times on the order of 0.2 0.7 s. The results suggest that the unsteady granular flow analysis may be beneficial for characterizing the "flowability" and "rheology" of granular materials based on particle size, moisture content, or other properties. PMID- 19905106 TI - Pulse propagation in tapered granular chains: an analytic study. AB - We study pulse propagation in one-dimensional tapered chains of spherical granules. Analytic results for the pulse velocity and other pulse features are obtained using a binary collision approximation. Comparisons with numerical results show that the binary collision approximation provides quantitatively accurate analytic results for these chains. PMID- 19905107 TI - Orientational ordering in sheared inelastic dumbbells. AB - Using even driven simulations, we show that homogeneously sheared inelastic dumbbells in two dimensions are randomly orientated in the limit of low density. As the packing fraction is increased, particles first tend to orient along the extensional axis, and then as the packing fraction is further increased, the alignment shifts closer to the flow axis. The orientational order parameter displays a continuous increase with packing fraction and does not appear to exhibit a universal scaling with elongation. Except at the highest packing fractions, the orientational distribution function can be reconstructed with only the first coefficient of the Fourier expansion. PMID- 19905108 TI - Physical test of a particle simulation model in a sheared granular system. AB - We report a detailed comparison of a slow gravity-driven sheared granular flow with a discrete-element simulation performed in the same geometry. In the experiments, grains flow inside a silo with a rectangular cross section and are sheared by a rough boundary on one side and smooth boundaries on the other sides. Individual grain position and motion are measured using a particle index-matching imaging technique where a fluorescent dye is added to the interstitial liquid which has the same refractive index as the glass beads. The simulations use a Cundall-Strack contact model between the grains using contact parameters that have been used in many other previous studies and ignore the hydrodynamic effects of the interstitial liquid. Computations are performed to understand the effect of particle coefficient of friction, elasticity, contact model, and polydispersity on mean flow properties. We then perform a detailed comparison of the particle fluctuation properties as measured by the displacement probability distribution function and the mean square displacement. All in all, our study suggests a high level of quantitative agreement between the simulations and experiments. PMID- 19905109 TI - Dilute wet granular particles: nonequilibrium dynamics and structure formation. AB - We investigate a gas of wet granular particles covered by a thin liquid film. The dynamic evolution is governed by two-particle interactions, which are mainly due to interfacial forces in contrast to dry granular gases. When two wet grains collide, a capillary bridge is formed and stays intact up to a certain distance of withdrawal when the bridge ruptures, dissipating a fixed amount of energy. A freely cooling system is shown to undergo a nonequilibrium dynamic phase transition from a state with mainly single particles and fast cooling to a state with growing aggregates such that bridge rupture becomes a rare event and cooling is slow. In the early stage of cluster growth, aggregation is a self-similar process with a fractal dimension of the aggregates approximately equal to Df approximately 2 . At later times, a percolating cluster is observed which ultimately absorbs all the particles. The final cluster is compact on large length scales, but fractal with Df approximately 2 on small length scales. PMID- 19905110 TI - Contact and stress anisotropies in start-up flow of colloidal suspensions. AB - Spatiotemporal correlations in start-up flows of attractive colloids are explored by numerical simulations as a function of their volume fraction and shear rate. The suspension is first allowed to flocculate during a time tw, then the stress necessary to induce its flow is computed. We find that, at low volume fractions, the stress is a universal function of the strain. On the contrary, at high volume fractions, this scaling behavior is no longer observed and a supplementary stress becomes necessary to induce flow. To better understand the physical origin of the supplementary stress, we examine the creation, disruption, and orientation of contacts between the particles and the corresponding contribution to stress as a function of strain. Our simulations show that the onset of flow is dominated by the creation of contacts between the particles at low shear rates and by their disruption at high shear rates. However, neither the evolution of the number of contacts with strain nor their orientation can fully account for the nonscaling behavior of the stress at high volume fractions. At small strains, the relative importance of forcing in the compression quadrant increases with volume fraction and with flocculation time. This mechanism of stress transmission through the compression quadrant is not accounted for in the usual description of yield stress, which considers the breaking of bonds oriented in the extension quadrant. PMID- 19905111 TI - Supersaturated dispersions of rodlike viruses with added attraction. AB - The kinetics of isotropic-nematic (I-N) and nematic-isotropic (N-I) phase transitions in dispersions of rodlike fd viruses are studied. Concentration quenches were applied using pressure jumps in combination with polarization microscopy, birefringence, and turbidity measurements. The full biphasic region could be accessed, resulting in the construction of an experimental analog of the bifurcation diagram. The N-I spinodal points for dispersions of rods with varying concentrations of depletion agent (dextran) were obtained from orientation quenches using cessation of shear flow in combination with small-angle light scattering. We found that the location of the N-I spinodal point is independent of the attraction, which was confirmed by theory. Surprisingly, the experiments showed that also the absolute induction time, the critical nucleus, and the growth rate are insensitive of the attraction if the concentration is scaled to the distance to the phase boundaries. PMID- 19905112 TI - Scattering information obtained by optical microscopy: differential dynamic microscopy and beyond. AB - We describe the use of a bright-field microscope for dynamic light scattering experiments on weakly scattering samples. The method is based on collecting a time sequence of microscope images and analyzing them in the Fourier space to extract the characteristic time constants as a function of the scattering wave vector. We derive a theoretical model for microscope imaging that accounts for (a) the three-dimensional nature of the sample, (b) the arbitrary coherence properties of the light source, and (c) the effect of the finite numerical aperture of the microscope objective. The model is tested successfully against experiments performed on a colloidal dispersion of small spheres in water, by means of the recently introduced differential dynamic microscopy technique [R. Cerbino and V. Trappe, Phys. Rev. Lett. 100, 188102 (2008)]. Finally, we extend our model to the class of microscopy techniques that can be described by a linear space-invariant imaging of the density of the scattering centers, which includes, for example, dynamic fluorescence microscopy. PMID- 19905113 TI - Ground state structures in ferrofluid monolayers. AB - A combination of analytical calculations and Monte Carlo simulations is used to find the ground state structures in monodisperse ferrofluid monolayers. Taking into account the magnetic dipole-dipole interaction between all particles in the system we observe different topological structures that are likely to exist at low temperatures. The most energetically favored structures we find are rings, embedded rings, and rings side by side, and we are able to derive analytical expressions for the total energy of these structures. A detailed analysis of embedded rings and rings side by side shows that the interring interactions are negligible. We furthermore find that a single ideal ring is the ground state structure for a ferrofluid monolayer. We compared our theoretical predictions to the results of simulated annealing data and found them to be in excellent agreement. PMID- 19905114 TI - Phase behavior of the hard-sphere Maier-Saupe fluid under spatial confinement. AB - The Maier-Saupe hard-sphere fluid is one of the simplest models that accounts for the isotropic-nematic transition characteristic of liquid crystal phases. At low temperatures the model is known to present a gas-liquid-like transition with a large difference between the densities of the coexistence phases, whereas at higher temperature the transition becomes a weak first-order transition resembling the typical order-disorder (nematic-isotropic) phase change of liquid crystals. Spatial dimensionality directly conditions the character of the orientational phase change (i.e., the high temperature transition), that goes from a first-order transition in the purely three-dimensional case, to a Berezinskii-Kosterlitz-Thouless-like continuous transition which occurs when the three dimensional Maier-Saupe spins are constrained to lie on a plane. In the latter instance, the ordered phase is not endowed with true long-range order. In this work we investigate how the continuous transition transforms into a true first-order phase change, by analyzing the phase behavior of a system of three dimensional Maier-Saupe hard spheres confined between two parallel plates, with separations ranging from the quasi-two-dimensional regime to the bulk three dimensional limit. Our results indicate that spatial confinement in one direction induces the change from first order to a continuous transition with a corresponding decrease of the transition temperatures. As to the gas-liquid transition, the estimated "critical" temperatures and densities also decrease as the fluid is confined, in agreement with previous results for other simple systems. PMID- 19905115 TI - Anomalous phase behavior of a soft-repulsive potential with a strictly monotonic force. AB - We study the phase behavior of a classical system of particles interacting through a strictly convex soft-repulsive potential which, at variance with the pairwise softened repulsions considered so far in the literature, lacks a region of downward or zero curvature. Nonetheless, such interaction is characterized by two length scales, owing to the presence of a range of interparticle distances where the repulsive force increases, for decreasing distance, much more slowly than in the adjacent regions. We investigate, using extensive Monte Carlo simulations combined with accurate free-energy calculations, the phase diagram of the system under consideration. We find that the model exhibits a fluid-solid coexistence line with multiple re-entrant regions, an extremely rich solid polymorphism with solid-solid transitions, and waterlike anomalies. In spite of the isotropic nature of the interparticle potential, we find that among the crystal structures in which the system can exist, there are also a number of non Bravais lattices, such as cI16 and diamond. PMID- 19905116 TI - Universality in multicomponent glass-forming liquids near the glass transition. AB - The slow dynamics of a single particle in multicomponent glass-forming systems including fragile and strong glasses is studied from a unified point of view. The simulation results on two different systems, bulk glass-forming Cu60Ti20Zr20 melt and network-forming SiO2 , melt are analyzed by the mean-field theory (MFT) recently proposed and are compared with other systems near the glass transition. It is shown that the simulation results for the mean-square displacement are all collapsed into a master curve given by MFT if a long-time self-diffusion coefficient has the same value in each system. It is also shown that each long time self-diffusion coefficient is described well by a singular function predicted recently from first principles. Thus, we conclude that there exists a simple universal mechanism near the glass transition even among any diversely different glass-forming systems. PMID- 19905117 TI - Self-organized dendritic sidebranching in directional solidification: sidebranch coherence within uncorrelated bursts. AB - We experimentally study the level of organization of dendritic sidebranching in directional solidification. For this, we extract successive interface positions at a fixed distance from the dendrite tips and we perform various correlation analyses. The sidebranching signals appear composed of randomly distributed bursts in which sidebranching coherence is surprisingly large and robust. This is attested by the large autocorrelation found in single bursts and the large cross correlation found in any couple of bursts, even belonging to different sides of a dendrite or to different dendrites. However, the phase coherence of sidebranching breaks down at the transition between bursts. This restricts the coherence of extended sidebranching signals to a mean burst length and prevents the occurrence of large scale cross-correlation between them. This balanced view on sidebranching coherence stresses the capability of self-organization of dendrites in material science and sheds light on the nature of sidebranching on curved growing forms. PMID- 19905118 TI - Thermodynamics of bcc metals in phase-field-crystal models. AB - We examine the influence of different forms of the free-energy functionals used in the phase-field-crystal (PFC) model, and compare them with the second-order density-functional theory (DFT) of freezing, by using bcc iron as an example case. We show that there are large differences between the PFC and the DFT and it is difficult to obtain reasonable parameters for existing PFC models directly from the DFT. Therefore, we propose a way of expanding the correlation function in terms of gradients that allows us to incorporate the bulk modulus of the liquid as an additional parameter in the theory. We show that this functional reproduces reasonable values for both bulk and surface properties of bcc iron, and therefore it should be useful in modeling bcc materials. As a further demonstration, we also calculate the grain boundary energy as a function of misorientation for a symmetric tilt boundary close to the melting transition. PMID- 19905119 TI - Quantized liquid density-functional theory for hydrogen adsorption in nanoporous materials. AB - We develop a finite-temperature quantized version of density-functional theory of atomic and molecular liquids (QLDFT). Following the Kohn-Sham partitioning of the free energy, we introduce a noninteracting reference fluid of particles obeying the Maxwell-Boltzmann statistics. The kinetic and potential energy of the reference fluid are evaluated exactly. All remaining contributions, including interactions between fluid particles and corrections due to the appropriate quantum statistics are subsumed by an excess (in electronic DFT called exchange correlation) functional. Two variants to approximate the excess functional are presented: the simplest local-interaction expression (LIE-0) avoids the direct calculation of interparticle interactions and includes them in the excess functional, which is parametrized to reproduce experimental equation of state of normal hydrogen. The more sophisticated LIE-1 approximation is based on the weighted local-density approximation and includes the explicit interparticle interaction potential as well as the local approximation of the excess functional, the latter being weighted by the average over a spherical environment to include nonlocal effects in an approximate way. We apply LIE-0 and LIE-1 to two benchmark systems, bulk fluid hydrogen and hydrogen in a slit pore, and compare it with classical molecular-dynamics simulations employing the same potential. Both functionals produce similar results for direct quantum effects in adsorption free energy. At the same time, LIE-1 also yields a reasonable description of the fluid structure and classical packing effects, which are not reproduced by LIE-0. The source code of our implementation of the LIE-QLDFT is distributed under the GNU public license and is included as a supporting material. PMID- 19905120 TI - Condensation-induced jumping water drops. AB - Water droplets can jump during vapor condensation on solid benzene near its melting point. This phenomenon, which can be viewed as a kind of micro scale steam engine, is studied experimentally and numerically. The latent heat of condensation transferred at the drop three phase contact line melts the substrate during a time proportional to R (the drop radius). The wetting conditions change and a spontaneous jump of the drop results in random direction over length approximately 1.5R , a phenomenon that increases the coalescence events and accelerates the growth. Once properly rescaled by the jump length scale, the growth dynamics is, however, similar to that on a solid surface. PMID- 19905121 TI - Freezing in the bulk controlled by prefreezing at a surface. AB - We use Monte Carlo simulations of the Lennard-Jones model to study the nucleation of a crystal phase at a flat surface. Our motivation is the observation that crystal phases almost always nucleate at a surface. We find that a surface phase transition (prefreezing) can control nucleation of the bulk crystal. This finding should be general and so surface phase behavior should be considered whenever nucleation of bulk phases at surfaces is considered. Also, nucleation of the bulk crystal transforms smoothly into the nucleation of a surface crystal layer as the bulk transition is crossed. PMID- 19905122 TI - Observation of local thickness fluctuations in surfactant membranes using neutron spin echo. AB - Experimental evidence of local thickness fluctuations of a surfactant membrane, as observed by neutron scattering, is reported. A swollen lamellar structure consisting of nonionic surfactant, water, and oil was investigated by neutron spin echo spectroscopy. Different dynamical processes are recognized at three different length scales. At length scales larger than the membrane thickness, the bending motion is observed, which follows the Zilman and Granek theory [Phys. Rev. Lett. 77, 4788 (1996)]. At the length scale corresponding to the membrane thickness, a clear excess dynamics in addition to the bending motion is observed. This mode is interpreted as the local thickness fluctuations. At even shorter length scales, smaller than the membrane thickness, intramembrane dynamics such as protrusion motions may have been observed. PMID- 19905123 TI - Understanding adsorption and desorption processes in mesoporous materials with independent disordered channels. AB - Using a lattice-gas model in mean-field theory, we discuss the problem of how adsorption and desorption of fluids in independent cylinderlike pores is influenced by variations in the pore diameter along the length of the pore, surface roughness of the pore walls, and chemical heterogeneity. We also consider the impact of contact with the bulk phase via the pore opening and the possibility of interactions between neighboring pores via a liquid film on the external surface of the material. We find that a combination of pore size variation along the length of the pore and surface roughness yields sorption hysteresis similar to that found in systems with three-dimensional disordered pore networks such as porous glasses. Our results are especially relevant to adsorption and desorption in porous silicon materials with independent linear pores and apparently anomalous features of the behavior in these systems can be accounted for within the context of the present model. PMID- 19905124 TI - Shear rate threshold for the boundary slip in dense polymer films. AB - The shear rate dependence of the slip length in thin polymer films confined between atomically flat surfaces is investigated by molecular dynamics simulations. The polymer melt is described by the bead-spring model of linear flexible chains. We found that at low shear rates the velocity profiles acquire a pronounced curvature near the wall and the absolute value of the negative slip length is approximately equal to the thickness of the viscous interfacial layer. At higher shear rates, the velocity profiles become linear and the slip length increases rapidly as a function of shear rate. The gradual transition from no slip to steady-state slip flow is associated with faster relaxation of the polymer chains near the wall evaluated from decay of the time autocorrelation function of the first normal mode. We also show that at high melt densities the friction coefficient at the interface between the polymer melt and the solid wall follows a power-law decay as a function of the slip velocity. At large slip velocities the friction coefficient is determined by the product of the surface induced peak in the structure factor, the temperature, and the contact density of the first fluid layer near the solid wall. PMID- 19905125 TI - Spontaneously chiral domains of an achiral bent-core nematic liquid crystal in a planar aligned device. AB - An achiral fixed bent-core nematic liquid crystal in an antiparallel aligned planar device is observed to form three distinct domains. Regions are observed which are achiral planar, as well as domains displaying both positive and negative chirality that are stable over a significant temperature interval. In such a device the presence of spontaneous positive and negatively twisted domains is highly unusual. By studying the electro-optics of the device relevant director structures are inferred, and it is additionally found that a sufficiently large applied electric field will drive the chiral domains into the achiral state. PMID- 19905126 TI - Mesogenic lattice models with partly antinematic interactions producing uniaxial nematic phases. AB - The present paper considers nematogenic lattice models, involving particles of D_{2h} symmetry, whose centers of mass are associated with a three-dimensional simple cubic lattice; the pair potential is isotropic in orientation space and restricted to nearest neighbors. Let two orthonormal triads define orientations of a pair of interacting particles; the simplest potential models proposed in the literature can be reduced to a linear combination involving the squares of the scalar products between corresponding unit vectors only and depending on three parameters. By now, various sets of potential parameters have been proposed and studied in the literature, some of which capable of producing biaxial orientational order at sufficiently low temperature. On the other hand, in experimental terms, mesogenic biaxial molecules mostly produce uniaxial mesophases; thus we address here two very simple cases, involving a nematic (calamitic) term as well as one (model P0M) or two (model PPM) antinematic ones, whose coefficients are set equal in magnitude; when only one antinematic coefficient is used, the third one is set to zero. The calamitic term favors the alignment of two corresponding molecular axes, whereas antinematic terms or geometric constraints tend to keep two other pairs of axes mutually orthogonal. The models were investigated by molecular-field treatments and Monte Carlo simulation and found to predict a first- or second-order transitions between uniaxial nematic and isotropic phases; the molecular-field treatments yielded results in reasonable agreement with simulation. PMID- 19905127 TI - Liquid-crystal transitions: a first-principles multiscale approach. AB - A rigorous theory of liquid-crystal transitions is developed starting from the Liouville equation. The starting point is an all-atom description and a set of order-parameter field variables that are shown to evolve slowly via Newton's equations. The separation of time scales between that of atomic collision or vibrations and the order-parameter fields enables the derivation of rigorous equations for stochastic order-parameter field dynamics. When the fields provide a measure of the spatial profile of the probability of molecular position, orientation, and internal structure, a theory of liquid-crystal transitions emerges. The theory uses the all-atom/continuum approach developed earlier to obtain a functional generalization of the Smoluchowski equation wherein key atomic details are embedded. The equivalent nonlocal Langevin equations are derived, and the computational aspects are discussed. The theory enables simulations that are much less computationally intensive than molecular dynamics and thus does not require oversimplification of the system's constituent components. The equations obtained do not include factors that require calibration and can thus be applicable to various phase transitions which overcomes the limitations of phenomenological field models. The relation of the theory to phenomenological descriptions of nematic and smectic phase transitions, and the possible existence of other types of transitions involving intermolecular structural parameters are discussed. PMID- 19905128 TI - Dispersions of pyrogenic alumina in pentylcyanobiphenyl studied by deuteron NMR. AB - Dispersions of hydrophilic (Aeroxide Alu C) and hydrophobic (Aeroxide Alu C 805) pyrogenic alumina (Al2O3) in liquid crystal 4;{'} -n-pentyl-4-cyanobiphenyl (5CB) were investigated with deuteron nuclear magnetic resonance. The disorder effects of Al2O3 particles on the orientational order of liquid-crystal media and on the field-induced director configuration were studied as a function of alumina density in samples prepared by zero-field cooling and in-field cooling procedures. The order parameters and their variation with alumina density suggest a stronger disordering effect from the nonpolar surface of Alu C 805 particles. For dispersions of hydrophobic Alu C 805 experiments involving in-field cooling from the isotropic phase indicate that the director of "disordered" domains can be aligned, though not perfectly, by the field-aided annealing process. But the same in-field cooling procedure has shown rather limited alignment effects for hydrophilic Alu C/5CB samples. The more robust network of hydrophilic gel possibly coupled with weak liquid-crystal-network interactions could be responsible for the observed behavior. Spectra recorded during in-field cooling and within the isotropic-nematic coexistence region reveal the augmentation of the disorder strength during the transition and illustrate the effect of field aided annealing. The stability of the aligned states as revealed by deuteron NMR is described. The results are discussed in comparison with previous studies of aerosil dispersions in alkylcyanobiphenyl. PMID- 19905129 TI - Variational theory for nematoacoustics. AB - The effect of an ultrasonic wave on the nematic texture has long been known, but its interpretation in terms of a coherent dynamical theory has not yet been achieved. A proposal for such a theory is made in this paper. The diverse theoretical approaches attempted in the past to describe the interaction between sound and nematic molecular orientation are briefly summarized. A theory for second-grade fluids, which provides the appropriate theoretical background for nematoacoustics, is also revived. An explicit application of the proposed theory to a simple computable case is given, which yields predictions that are qualitatively confirmed by a number of experimental results. PMID- 19905130 TI - Simulation of cholesteric blue phases using a Landau-de Gennes theory: effect of an applied electric field. AB - We investigate numerically static and dynamic properties of cholesteric blue phases. Our study is based on a Landau-de Gennes theory describing the orientational order of a liquid crystal in terms of a second-rank tensor. To find the shape and size of the unit cell conforming to the minimum of the free energy, we let the geometrical parameters characterizing the unit cell relax in the course of the time evolution via a simple relaxational equation. We investigate the effect of an electric field on the structure of cholesteric blue phases. We study how the deformation of the unit cell in response to the electric field E depends on the strength and direction of the electric field and the original structure of cholesteric blue phases. Our results qualitatively agree with the experimental findings. Although in a weak field, the strain tensor is proportional to E2 as previously argued, for a moderate field the distortion is no longer proportional to E2 and can be even nonmonotonic with respect to E2 . Furthermore, we investigate the kinetic processes of the deformation, rearrangement, and extinction of disclination lines under a strong electric field. We show that the kinetics of disclination lines is highly complicated and sensitively depends on the initial structure of blue phases, the direction of the electric field, and the sign of dielectric anisotropy epsilon(a). In most cases, a strong field aligns the liquid crystals in a uniform (positive epsilon(a)) or helical (negative epsilon(a)) manner without disclination lines. However, for negative epsilon(a) and the direction of the electric field parallel to the body diagonal of the unit cell, disclination lines do not disappear and form a two dimensional hexagonal lattice. PMID- 19905131 TI - Comparison between experiment and theory in the temperature variation of film tension above the bulk isotropic transition in free-standing liquid-crystal films. AB - Using differential scanning calorimetry, the transition enthalpies and temperatures for the bulk smectic-isotropic phase transition have been measured for a series of liquid-crystal compounds. For five compounds, those values were used as parameters in a microscopic mean-field model to predict the temperature dependence of the difference in free-energy density between a sample of material in a free-standing smectic film and that in the bulk. The model predicts a weak temperature dependence below the bulk clearing point and a pronounced monotonic increase with temperature above the transition temperature. The compounds used in this study were chosen specifically because they were also the subject of a previous independent experimental study [M. Veum, Phys. Rev. E 74, 011703 (2006)] that demonstrated a sudden monotonic increase in the free-standing film tension with temperature, which is qualitatively consistent with the predictions of the above-mentioned mean-field model. This study presents a direct and quantitative comparison between the predictions of the mean-field model and the results from previous tension experiments. PMID- 19905132 TI - Liquid-crystal pumping in a cylindrical capillary with radial temperature gradient. AB - Dynamic field pumping principle has been developed utilizing the interactions of both the director and velocity fields and a temperature gradient inverted deltaT. The orientational dynamics in the hybrid-oriented liquid-crystal (HOLC) microvolume confined between two infinitely long coaxial cylinders under the influence of the radially directed inverted deltaT has been investigated. We have carried out a numerical study of a system of hydrodynamic equations including director reorientation, fluid flow, and temperature redistribution across the HOLC cavity between two cylinders under the influence of inverted deltaT, when the liquid-crystal cavity is heated both from outer (inner) to inner (outer) bounding cylinders. Calculations show that under the influence of inverted deltaT the initially quiescent HOLC drop settles down to a stationary flow regime, with the horizontal u(st)(r) component of velocity. The effects of inverted deltaT , of the character of the preferred anchoring of the average molecular direction to the restricted cylinders, and of the size of the HOLC cavity on magnitude and direction of hydrodynamic flow--for a number of hydrodynamic regimes--has been investigated. PMID- 19905133 TI - Flexoelectro-optic properties of chiral nematic liquid crystals in the uniform standing helix configuration. AB - The flexoelectro-optic effect describes the rotation of the optic axis of a short pitch chiral nematic liquid crystal under the application of an electric field. We investigate the effect in the uniform standing helix, or "Grandjean" configuration. An in-plane electric field is applied. The director profile is determined numerically using a static one-dimensional continuum model with strong surface anchoring. The Berreman method is used to solve for plane-wave solutions to Maxwell's equations, and predict the optical properties of the resulting structure in general cases. By using a chiral nematic with short pitch between crossed polarizers an optical switch may be generated. With no applied field the configuration is nontransmissive at normal incidence, but becomes transmissive with an applied field. For this case, numerical results using the Berreman method are supplemented with an analytic theory and found to be in good agreement. The transmitted intensity as a function of tilt, the contrast ratio, and the tilt required for full intensity modulation are presented. The angular dependence of the transmission is calculated and the isocontrast curves are plotted. For typical material and cell parameters a switching speed of 0.017 ms and contrast ratio of 1500:1 at normal incidence are predicted, at a switch-on tilt of 41.5 degrees. Experimental verification of the analytic and numerical models is provided. PMID- 19905134 TI - Mechanically induced Helfrich-Hurault effect in lamellar systems. AB - Layered phases are a common pattern of self-organization for several soft materials. These phases undergo buckling instability when subjected to dilatative strain: beyond a critical threshold, layers, initially flat, exhibit a periodical undulation. By using a continuum model, in a finite deformation framework, an expression for the critical threshold is provided, which differs from that predicted by the Helfrich-Hurault theory and yet it reverts to it in a thick specimen limit. With respect to the relevant literature, an analogous disagreement is found in the undulation amplitude expression as well. The obtained results appear particularly relevant when dealing with layered materials whose intrinsic coherence length is comparable to the cell thickness. PMID- 19905135 TI - Director reorientation in a hybrid-oriented liquid-crystal film induced by thermomechanical effect. AB - We have carried out a numerical study of a system of hydrodynamic equations including director reorientation, fluid flow, and temperature redistribution across a two-dimensional (2D) hybrid-oriented liquid-crystal (HOLC) cell under the influence of a heat flow directed normal to the upper bounding surface, whereas on the rest boundaries the temperature is kept constant. Calculations based upon the nonlinear extension of the classical Ericksen-Leslie theory shows that the HOLC material under the influence of the heat flow, after some time, more than the time of relaxation, for instance, of the director field in the HOLC cell, settles down to the rest state regime, where the horizontal and vertical components of the velocity vector are equal to zero, and the temperature field across the LC cell finally reaches the value of temperature on the lower and two lateral bounding surfaces. The role of hydrodynamic flow in the relaxation processes of the temperature field to its equilibrium distribution across the 2D HOLC cell, containing 4-n-pentyl-4'-cyanobiphenyl, has been investigated, for a number of dynamic regimes. PMID- 19905136 TI - Extension of the Hamaneh-Taylor model using the macroscopic polarization for the description of chiral smectic liquid crystals. AB - Chiral smectic liquid crystals exhibit a series of phases, including ferroelectric, antiferroelectric, and ferrielectric commensurate structures as well as an incommensurate Sm-Calpha* phase. We carried out an extension of the phenomenological model recently presented by Hamaneh and Taylor based on the distorted-clock model. The salient feature of this model is that it links the appearance of phases to a spontaneous microscopic twist: i.e., an increment alpha of the azimuthal angle from layer to layer. The balance between this twist and an orientational order parameter J gives the effective phase. We introduce a second orientational order parameter I , which physical meaning comes from the macroscopic polarization; the effect of an applied electric is also studied. We derive phase diagrams and correlate them to our experimental results under field showing the sequence of phases versus temperature and electric field in some compounds. PMID- 19905137 TI - Gel-like elasticity in glass-forming side-chain liquid-crystal polymers. AB - We study the complex shear modulus G of two side-chain liquid-crystal polymers (SCLCPs), a methoxy-phenylbenzoate substituted polyacrylate (thereafter called PAOCH3 ), and a cyanobiphenyl substituted polyacrylate supplied by Merck (thereafter called LCP105) using a piezoelectric rheometer. Two methods of filling the cell are used: (a) a capillary method, which can be used only at high temperature because of the low value of the viscosity, and (b) the classical one, thereafter called compression method, which consists in placing the sample between the two slides of the cell and to bring them closer. By filling the cell at high temperature either with the compression or the capillary method, we show that the response of both compounds is liquidlike ( G' approximately f2 and G'' approximately f , where f is the frequency) for temperatures higher than a certain temperature T0 and gel-like (G' approximately const, G'' approximately f) below T0. This change in behavior from the conventional flow response to a gel like response, when approaching the glass transition, is observed for nonsliding conditions and for very weak-imposed shear strains. It can be explained by a percolation-type mechanism of preglassy elastic clusters, which correspond to long-range and long-lived density fluctuations that are frozen at the time scale of the experiment. The sample response is therefore the sum of two contributions: one is due to the flow response of the polymer melt and the other to the elastic response of the network formed by the preglassy elastic clusters. By filling the cell below T0 with the compression method, both compounds exhibit a gel-type behavior by gently bringing closer the slides of the cell and an anomalous low frequency behavior characterized by G'=const and G''=const by increasing the pressure used to bring closer the slides of the cell. A compression-assisted aggregation of the preglassy elastic clusters can explain both the increase in the low-frequency elastic plateau when the sample thickness is decreased and the anomalous low-frequency behavior. Further evidence for the existence of these elastic clusters is provided by the following results: (a) the nonlinear response of the samples as a function of the strain amplitude, which can be explained by the Payne effect, and (b) the aggregation effects, which can be mimicked by a polydimethylsiloxane melt filled with silica particles, the silica particles playing the role of the preglassy elastic clusters. All these observations show that PAOCH3 is not a macroscopically solidlike material with an unconventional type of elasticity, as claimed by Mendil [Phys. Rev. Lett. 96, 077801 (2006)]. The gel-type behavior observed here on two SCLCPs ( PAOCH3 and LCP105) and previously on some conventional flexible polymers (atactic polystyrene, poly-n butylacrylate) seems to be a generic effect of the glass transition. The presence of the preglassy elastic clusters questions the widely accepted hypothesis of ergodicity in the supercooled state. PMID- 19905138 TI - Glass transition and molecular mobility in polymer thin films. AB - Extensive studies on polymer thin films to date have revealed their interesting but unusual properties such as film thickness dependence of glass transition temperature Tg and thermal expansivity. Recent studies have shown that the lower Tg is not always related to the higher mobility in polymer thin films, which contradicts our current understanding of the glass transition process. In this work, we report the results of inelastic neutron-scattering measurements on polystyrene thin films using two spectrometers with different energy resolutions as well as ellipsometry measurements. The results are interpreted in terms of cooperatively rearranging region and motional slowing down due to the surface effect that explain plausibly the anomalous relationship between the glass transition temperature Tg and the molecular mobility in thin films. PMID- 19905139 TI - Topological analysis of polymeric melts: chain-length effects and fast-converging estimators for entanglement length. AB - Primitive path analyses of entanglements are performed over a wide range of chain lengths for both bead spring and atomistic polyethylene polymer melts. Estimators for the entanglement length N_{e} which operate on results for a single chain length N are shown to produce systematic O(1/N) errors. The mathematical roots of these errors are identified as (a) treating chain ends as entanglements and (b) neglecting non-Gaussian corrections to chain and primitive path dimensions. The prefactors for the O(1/N) errors may be large; in general their magnitude depends both on the polymer model and the method used to obtain primitive paths. We propose, derive, and test new estimators which eliminate these systematic errors using information obtainable from the variation in entanglement characteristics with chain length. The new estimators produce accurate results for N_{e} from marginally entangled systems. Formulas based on direct enumeration of entanglements appear to converge faster and are simpler to apply. PMID- 19905140 TI - Budding and vesiculation induced by conical membrane inclusions. AB - Conical inclusions in a lipid bilayer generate an overall spontaneous curvature of the membrane that depends on concentration and geometry of the inclusions. Examples are integral and attached membrane proteins, viruses, and lipid domains. We propose an analytical model to study budding and vesiculation of the lipid bilayer membrane, which is based on the membrane bending energy and the translational entropy of the inclusions. If the inclusions are placed on a membrane with similar curvature radius, their repulsive membrane-mediated interaction is screened. Therefore, for high inclusion density the inclusions aggregate, induce bud formation, and finally vesiculation. Already with the bending energy alone our model allows the prediction of bud radii. However, in case the inclusions induce a single large vesicle to split into two smaller vesicles, bending energy alone predicts that the smaller vesicles have different sizes whereas the translational entropy favors the formation of equal-sized vesicles. Our results agree well with those of recent computer simulations. PMID- 19905141 TI - Isotropic-nematic phase transition in amyloid fibrilization. AB - We carry out a theoretical study on the isotropic-nematic phase transition and phase separation in amyloid fibril solutions. Borrowing the thermodynamic model employed in the study of cylindrical micelles, we investigate the variations in the fibril length distribution and phase behavior with respect to changes in the protein concentration, fibril's rigidity, and binding energy. We then relate our theoretical findings to the nematic ordering experimentally observed in Hen Lysozyme fibril solution. PMID- 19905142 TI - Enhanced mixing and spatial instability in concentrated bacterial suspensions. AB - High-resolution optical coherence tomography is used to study the onset of a large-scale convective motion in free-standing thin films of adjustable thickness containing suspensions of swimming aerobic bacteria. Clear evidence is found that beyond a threshold film thickness there exists a transition from quasi-two dimensional collective swimming to three-dimensional turbulent behavior. The latter state, qualitatively different from bioconvection in dilute bacterial suspensions, is characterized by enhanced diffusivities of oxygen and bacteria. These results emphasize the impact of self-organized bacterial locomotion on the onset of three-dimensional dynamics, and suggest key ingredients necessary to extend standard models of bioconvection to incorporate effects of large-scale collective motion. PMID- 19905143 TI - Continuous model for microtubule dynamics with catastrophe, rescue, and nucleation processes. AB - Microtubules are a major component of the cytoskeleton distinguished by highly dynamic behavior both in vitro and in vivo referred to as dynamic instability. We propose a general mathematical model that accounts for the growth, catastrophe, rescue, and nucleation processes in the polymerization of microtubules from tubulin dimers. Our model is an extension of various mathematical models developed earlier formulated in order to capture and unify the various aspects of tubulin polymerization. While attempting to use a minimal number of adjustable parameters, the proposed model covers a broad range of behaviors and has predictive features discussed in the paper. We have analyzed the range of resultant dynamical behavior of the microtubules by changing each of the parameter values at a time and observing the emergence of various dynamical regimes that agree well with the previously reported experimental data and behavior. PMID- 19905144 TI - Structural variability and the incoherent addition of scattered intensities in single-particle diffraction. AB - X-ray lasers may allow structural studies on single particles and biomolecules without crystalline periodicity in the samples. We examine here the effect of sample dynamics as a source of structural heterogeneity on the resolution of the reconstructed image of a small protein molecule. Structures from molecular dynamics simulations of lysozyme were sampled and aligned. These structures were then used to calculate diffraction patterns corresponding to different dynamic states. The patterns were incoherently summed and the resulting data set was phased using the oversampling method. Reconstructed images of hydrated and dehydrated lysozyme gave resolutions of 3.7 A and 7.6 A , respectively. These are significantly worse than the root-mean-square deviation of the hydrated ( 2.7 A for all atoms and 1.45 A for C-alpha positions) or dehydrated ( 3.7 A for all atoms and 2.5 A for C-alpha positions) structures. The noise introduced by structural dynamics and incoherent addition of dissimilar structures restricts the maximum resolution to be expected from direct image reconstruction of dynamic systems. A way of potentially reducing this effect is by grouping dynamic structures into distinct structural substates and solving them separately. PMID- 19905145 TI - Interaction of electrically evoked responses in networks of dissociated cortical neurons. AB - In this work we describe the interaction of the responses of neuronal networks to pairs of electrical stimuli. For this we use networks of dissociated cortical neurons cultured on planar microelectrode arrays. We compare the response to pairs of stimuli with the response to a stimulus in isolation. To evaluate the influence of both stimuli we introduce a normalization of the root-mean square of the response. Furthermore we consider the response to a pair of stimuli as the linear superposition of the two constituents. The two methods combined show that the neuronal network strongly suppresses the second stimulus. At the same time we find a possible window of integration in which two stimuli from separate locations in the network can interact to form a new response. PMID- 19905146 TI - Inhibition and modulation of rhythmic neuronal spiking by noise. AB - We investigated the effects of noise on periodic firing in the Hodgkin-Huxley nonlinear system. With mean input current mu as a bifurcation parameter, a bifurcation to repetitive spiking occurs at a critical value microc approximately 6.44 . The firing behavior was studied as a function of the mean and variance of the input current, firstly with initial resting conditions. Noise of a small amplitude can turn off the spiking for values of micro close to microc, and the number of spikes undergoes a minimum as a function of the noise level. The robustness of these phenomena was confirmed by simulations with random initial conditions and with random time of commencement of the noise. Furthermore, their generality was indicated by their occurrence when additive noise was replaced by conductance-based noise. For long periods of observation, many frequent transitions may occur from spiking to nonspiking activity when the noise is sufficiently strong. Explanations of the above phenomena are sought in terms of the underlying bifurcation structure and the probabilities that noise shifts the process from the basin of attraction of a stable limit cycle to that of a stable rest state. The waiting times for such transitions depend strongly on the values of mu and sigma and on the forms of the basins of attraction. The observed effects of noise are expected to occur in diverse fields in systems with the same underlying dynamical structure. PMID- 19905147 TI - Fluctuation dynamics of spherical vesicles: frustration of regular bulk dissipation into subdiffusive relaxation. AB - Spherical lipid vesicles obtained by the extrusion method are nonequilibrium membrane structures more curved than the zero spontaneous curvature equilibrium state of the bilayer. Furthermore, these structures are quite rigid as compared to spontaneous vesicles or microemulsion droplets made of soluble surfactants. The dynamical description of the shape fluctuations derived by Milner and Safran (MS) [Phys. Rev. A 36, 4371 (1987)], which is based on the elastic Helfrich energy referred to the equilibrium state, could be misleading in these cases. In the present contribution, shape fluctuations of unilamellar palmitoyl-oleyl phosphocholine (POPC) vesicles (radius Rh