Diet Supplemented with Chia Flour did not Modified the Inflammatory Process and Tumor Development in Wistar Rats Inoculated with Walker 256 Cells.

Chia seed (Salvia hispanica L.) contains high amounts of n-3 α-linolenic acid (ALA) and has been associated with many health benefits. The aim of the present study was to evaluate the AIN-93 diet supplemented by chia flour on cancer-cachexia development and tissues inflammatory response. Wistar rats at 30 days old were treated with control diet or diet supplemented with chia flour for eight weeks. After this period, half of the animals in each diet group were inoculated with Walker 256 tumor cells. On the 14th day after tumor inoculation, the animals were euthanized and white adipose tissue depots, liver, gastrocnemius muscle, and tumor were removed. The tumor weight was higher and IL-10 content was lower in chia flour group. The tumor bearing did not modify the cytokines content in gastrocnemius muscle, retroperitoneal and epididymal adipose tissue, however, it decreased IL-1ß and TNF-α content in liver, and IL6R and IL-10R protein content in mesenteric adipose tissue. In conclusion, our results demonstrated that supplementation with chia flour did not prevent the tumor bearing effects in Walker 256 model.

Identification of a role for CLASP2 in insulin action.

Insulin stimulates the mobilization of glucose transporter 4 (GLUT4) storage vesicles to the plasma membrane, resulting in an influx of glucose into target tissues such as muscle and fat. We present evidence that CLIP-associating protein 2 (CLASP2), a protein previously unassociated with insulin action, is responsive to insulin stimulation. Using mass spectrometry-based protein identification combined with phosphoantibody immunoprecipitation in L6 myotubes, we detected a 4.8-fold increase of CLASP2 in the anti-phosphoserine immunoprecipitates upon insulin stimulation. Western blotting of CLASP2 immunoprecipitates with the phosphoantibody confirmed the finding that CLASP2 undergoes insulin-stimulated phosphorylation, and a number of novel phosphorylation sites were identified. Confocal imaging of L6 myotubes revealed that CLASP2 colocalizes with GLUT4 at the plasma membrane within areas of insulin-mediated cortical actin remodeling. CLASP2 is responsible for directing the distal end of microtubules to the cell cortex, and it has been shown that GLUT4 travels along microtubule tracks. In support of the concept that CLASP2 plays a role in the trafficking of GLUT4 at the cell periphery, CLASP2 knockdown by siRNA in L6 myotubes interfered with insulin-stimulated GLUT4 localization to the plasma membrane. Furthermore, siRNA mediated knockdown of CLASP2 in 3T3-L1 adipocytes inhibited insulin-stimulated glucose transport. We therefore propose a new model for CLASP2 in insulin action, where CLASP2 directs the delivery of GLUT4 to cell cortex landing zones important for insulin action.

Novel nutraceutical supplements with yeast ß-glucan, prebiotics, minerals, and Silybum marianum (silymarin) ameliorate obesity-related metabolic and clinical parameters: A double-blind randomized trial.

Purpose: It is known that obesity has a multifactorial etiology that involves genetic and environmental factors. The WHO estimates the worldwide prevalence of 1.9 billion overweight adults and more than 650 million people with obesity. These alarming data highlight the high and growing prevalence of obesity and represent a risk factor for the development and aggravation of other chronic diseases, such as nonalcoholic fatty liver disease (NAFLD) that is frequently considered the hepatic outcome of type 2 diabetes. The use of non-pharmacological therapies such as food supplements, nutraceuticals, and natural integrative therapies has grown as an alternative tool for obesity-related diseases compared to conventional medications. However, it is a still little explored research field and lacks scientific evidence of therapeutic effectiveness. Considering this, the aim is to evaluate whether a new nutraceutical supplement composition can improve and supply essential mineral nutrients, providing an improvement of obesity-related metabolic and endocrine parameters. Methods: Sedentary volunteers (women and men) with body mass index (BMI) ≤34.9 kg/m2 were divided into two groups: Novel Nutraceutical Supplement_(S) (n = 30) and Novel Nutraceutical Supplement (n = 29), differing in the absence (S) or presence of silymarin, respectively. Volunteers were instructed to take two capsules in the morning and two capsules in the evening. No nutritional intervention was performed during the study period. The data (anthropometrics and anamneses) and harvest blood (biochemistry and hormonal exams) were collected at three different time points: baseline time [day 0 (T0)], day 90 (T90), and day 180 (T180) post-supplementation. Results: In the anthropometric analysis, the waist circumference in middle abdomen (WC-mid) and waist circumference in iliac crest (WC-IC) were reduced. Also, the waist-to-height ratio (WHt R) and waist-to-hip ratio (WHR) seem to slightly decrease alongside the supplementation period with both nutraceutical supplements tested as well as transaminase enzyme ratio [aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio (AAR)], a known as a biomarker of NAFLD, and endocrine hormones cortisol and thyroid-stimulating hormone (TSH) at 90 and 180 days post-supplementation. Conclusions: In a condition associated with sedentary and no nutritional intervention, the new nutraceutical supplement composition demonstrated the ability to be a strong and newfangled tool to improve important biomarkers associated with obesity and its comorbidities.

The Symbiotic Effect of a New Nutraceutical with Yeast ß-Glucan, Prebiotics, Minerals, and Silybum marianum (Silymarin) for Recovering Metabolic Homeostasis via Pgc-1α, Il-6, and Il-10 Gene Expression in a Type-2 Diabetes Obesity Model.

The use of natural products and derivatives for the prevention and control of non-communicable chronic diseases, such as type-2 diabetes (T2D), obesity, and hepatic steatosis is a way to achieve homeostasis through different metabolic pathways. Thus, male C57BL/6 mice were divided into the following groups: high-fat diet (HFD) vehicle, HFD + Supplemented, HFD + Supplemented_S, and isolated compounds. The vehicle and experimental formulations were administered orally by gavage once a day over the four weeks of the diet (28 consecutive days). We evaluated the energy homeostasis, cytokines, and mitochondrial gene expression in these groups of mice. After four weeks of supplementation, only the new nutraceutical group (HFD + Supplemented) experienced reduced fasting glycemia, insulin, HOMA index, HOMA-ß, dyslipidemia, ectopic fat deposition, and hepatic fibrosis levels. Additionally, the PPARγ coactivator 1 α (Pgc-1α), interleukin-6 (Il-6), and interleukin-10 (Il-10) gene expression were augmented, while hepatic steatosis decreased and liver parenchyma was recovered. The glutathione-S-transferase activity status was found to be modulated by the supplement. We discovered that the new nutraceutical was able to improve insulin resistance and hepatic steatosis mainly by regulating IL-6, IL-10, and Pgc-1α gene expression.

A novel supplement with yeast ß-glucan, prebiotic, minerals and Silybum marianum synergistically modulates metabolic and inflammatory pathways and improves steatosis in obese mice.

OBJECTIVE: To evaluate the synergic effects of a novel oral supplement formulation, containing prebiotics, yeast ß-glucans, minerals and silymarin (Silybum marianum), on lipid and glycidic metabolism, inflammatory and mitochondrial proteins of the liver, in control and high-fat diet-induced obese mice. METHODS: After an acclimation period, 32 male C57BL/6 mice were divided into the following groups: nonfat diet (NFD) vehicle, NFD supplemented, high-fat diet (HFD) vehicle and HFD supplemented. The vehicle and experimental formulation were administered orally by gavage once a day during the last four weeks of the diet (28 consecutive days). We then evaluated energy homeostasis, inflammation, and mitochondrial protein expression in these groups of mice. RESULTS: After four weeks of supplementation, study groups experienced reduced glycemia, dyslipidemia, fat, and hepatic fibrosis levels. Additionally, proliferator-activated receptor-α, AMP-activated protein kinase-1α, peroxisome proliferator-activated receptor γ co-activator-1α, and mitochondrial transcription factor A expression levels were augmented; however, levels of inhibitor of nuclear factor-κB kinase subunit α and p65 nuclear factor-κB expression, and oxidative markers were reduced. Notably, the cortisol/C-reactive protein ratio, a well-characterized marker of the hypothalamic-pituitary-adrenal axis immune interface status, was found to be modulated by the supplement. CONCLUSION: We discovered that the novel supplement was able to modify different antioxidant, metabolic and inflammatory pathways, improving the energy homeostasis and inflammatory status, and consequently alleviated hepatic steatosis.

A diet including xanthan gum triggers a pro-inflammatory response in Wistar rats inoculated with Walker 256 cells.

OBJECTIVE: The aim of this study was to evaluate the effect of adding xanthan gum to the diet of rats on the production of cytokines and pro-inflammatory factors and on tumor development in rats inoculated with Walker 256 tumor cells. METHODS: Fifty-six rats were divided into 4 groups: control diet (C), control diet with tumor (TC), xanthan gum diet (XG), xanthan gum diet with tumor (TXG). RESULTS: The ingestion of xanthan gum promotes changes in cytokine content: increasing IL-6 TNF-α and IL-10 in retroperitoneal adipose tissue compared to the control group; and increasing TNF-α in the mesenteric adipose tissue compared to the C and TXG groups. On the contrary, the addition of xanthan gum to the diet did not affect the development of Walker 256 tumors in rats. CONCLUSION: The continuous use of xanthan gum triggered a pro-inflammatory response, promoting an increase in pro-inflammatory cytokines in the adipose tissue, but it did not have an effect on the tumor development in the animals inoculated with Walker 256 tumor cells.

Chia flour (Salvia hispanica L.) did not improve the deleterious aspects of hyperlipidic diet ingestion on glucose metabolism, but worsened glycaemia in mice.

Obesity is mainly caused by intake of a high-fat diet and sedentarism, and is considered a public health issue worldwide. Increased intestinal permeability may favour endotoxaemia generated by lipopolysaccharides, a substance present in the cell membrane of Gram-negative bacteria, and, consequently, an increase in systemic inflammation and metabolic diseases. In contrast (On the other hand), consumption of a healthy diet can help in the prevention and treatment of metabolic syndrome. In this way, chia seeds (Salvia hispanica L.), rich in polyunsaturated fatty acids, may present an anti-inflammatory role. In addition, chia is rich in antioxidants like caffeic and gallic acid and fiber. However, few studies have investigated the relationship between chia seeds, inflammatory mechanisms and intestinal permeability. Therefore, the aim of this study was to analyse the effects of chia administration on metabolism in obese mice. Swiss mice were fed a hyperlipidic diet either supplemented with or without 3% chia flour for 16 weeks. The results showed that supplementation could not reduce the deleterious effects of the lipid-rich diet in terms of body composition, glucose intolerance and activity of antioxidants enzymes in the liver. In addition, supplementation with chia in the control diet decreased the amount of occludin in the intestinal colon. In conclusion, although chia did not improve metabolic parameters it seemed to restore the intestinal barriers integrity. The beneficial effects of chia seem to be dependent of the quantity used, since our data conflict with those in the literature; however, it is important to note that other studies, unlike our protocol, used chia in the form of seeds or oil, and not flour.

"Give What the People Want": A Situational Analysis of Condom Distribution and a Feasibility Study of User-Friendly Condoms in Cape Town, South Africa.

Condom use has remained low in South Africa despite efforts by the government to make condoms accessible to all. We assessed condom availability, procurement/distribution, and education practices, and explored perceptions of and interest in an expanded array of condom options, including fitted male condoms in Cape Town, South Africa. We conducted 11 key informant interviews with staff members from two sexual health organizations and four state-owned clinics. Key informants (a) were interested in their organizations offering an expanded variety of male condom options including different sizes, colors, and flavors; and (b) substantiated the capacity that their organizations had to integrate alternative condom options into existing procurement, distribution, and education programs. Challenges discussed included securing funding, educating clients, and addressing logistical issues of stocking and distributing new condom types. Further research should explore whether providing condoms with a greater array of characteristics could improve condom use in South Africa.

A influência da dopamina nos transtornos de depressão: revisão de literatura / The influence of dopamine on depression disorders: review of literature / La influencia de la dopamina en los transportes de la depresión: revisión de la literatura

A depressão é definida como um distúrbio que provoca alterações de ordem social, psicológica, fisiológica e biológica. Pessoas diagnosticadas como depressivas apresentam impactos no funcionamento psicossocial, saúde física, mortalidade e qualidade de vida. Por esta razão, este estudo teve como objetivo demonstrar os mecanismos fisiológicos envolvidos nos transtornos de depressão e sua relação com o neurotransmissor dopamina. Trata-se de uma revisão bibliográfica narrativa realizada no período de março a maio de 2023, através de pesquisas nas bases de dados Scientific Electronic Library Online (SciELO), PubMed via Medical Literature Analysis and Retrieval System Online (MEDLINE), Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS) e Google Scholar. Através desta revisão foi possível evidenciar a importância no conhecimento acerca dos mecanismos fisiopatológicos envolvidos nos transtornos de depressão, sendo essencial para o entendimento e aplicação em tratamentos de pacientes diagnosticados com este transtorno.

Depression is defined as a disorder that causes social, psychological, physiological and biological changes. People diagnosed as depressive have an impact on psychosocial functioning, physical health, mortality, and quality of life. For this reason, this study aimed to demonstrate the physiological mechanisms involved in depression disorders and their relationship with the neurotransmitter dopamine. It is a narrative bibliographic review conducted in the period from March to May 2023, through searches in the databases Scientific Electronic Library Online (SciELO), PubMed via Medical Literature Analysis and Retrieval System Online (MEDLINE), Latin American and Caribbean Literature in Health Sciences (LILACS) and Google Scholar. Through this review it was possible to highlight the importance in the knowledge about the physiopathological mechanisms involved in depression disorders, being essential for the understanding and application in treatments of patients diagnosed with this disorder.

La depresión se define como un trastorno que causa cambios sociales, psicológicos, fisiológicos y biológicos. Las personas diagnosticadas como depresivas tienen un impacto en el funcionamiento psicosocial, la salud física, la mortalidad y la calidad de vida. Por esta razón, este estudio tuvo como objetivo demostrar los mecanismos fisiológicos involucrados en los trastornos de la depresión y su relación con el neurotransmisor dopamina. Esta es una revisión bibliográfica narrativa realizada entre marzo y mayo de 2023, a través de la investigación en las bases de datos Scientific Electronic Library Online (SciELO), PubMed via Medical Literature Analysis and Retrieval System Online (MEDLINE), Latin American and Caribbean Literature in Health Sciences (LILACS) y Google Scholar. A través de esta revisión, se pudo destacar la importancia en el conocimiento de los mecanismos fisiopatológicos involucrados en los trastornos de la depresión, y es esencial para entenderlos y aplicarlos al tratamiento de pacientes diagnosticados con este trastorno.

Characteristics of keratoconus and pellucid marginal degeneration in mean curvature maps.

PURPOSE: To characterize keratoconus and pellucid marginal degeneration (PMD) on mean curvature maps. DESIGN: Observational case series. METHODS: A retrospective analysis of clinical records of 19 keratoconus eyes and eight PMD eyes. Keratometric mean curvature maps were computed from a placido-disk-based corneal topography system. The peak location and amplitude of distortions were quantified by fitting two-dimensional Gaussian functions. Computer simulations of keratoconic corneal topography were used to help interpret the results. RESULTS: Keratoconus and PMD eyes had a characteristic localized increase in convexity (cone) on mean curvature maps. Computer simulations showed that the peak on the mean curvature map accurately represented the peak of the cone-like distortion. However, the peak location and appearance of the same conic distortion on axial and tangential maps were greatly influenced by coexisting astigmatism. The Gaussian function provided an excellent model of keratoconus on mean curvature maps, with a mean cross-correlation of 0.80. According to Gaussian fitting, the vertical cone peak locations were -1.10 +/- 0.43 mm (mean +/- SD) in keratoconus eyes and -1.94 +/- 0.53 mm in PMD eyes (P < .01). The locations of peaks on axial and tangential maps differed considerably from those on mean curvature maps. Advanced keratoconus tended to have higher cones (amplitude 13.31 +/- 6.87 diopters) than early keratoconus (amplitude 10.65 +/- 1.56 diopters). CONCLUSIONS: Gaussian fitting of mean curvature maps accurately quantifies the peak location and amplitude of the cone in keratoconus and PMD eyes. This new topographic analysis might be useful in the diagnosis and tracking of corneal ectatic diseases.

Gene and MicroRNA Expression Responses to Exercise; Relationship with Insulin Sensitivity.

BACKGROUND: Healthy individuals on the lower end of the insulin sensitivity spectrum also have a reduced gene expression response to exercise for specific genes. The goal of this study was to determine the relationship between insulin sensitivity and exercise-induced gene expression in an unbiased, global manner. METHODS AND FINDINGS: Euglycemic clamps were used to measure insulin sensitivity and muscle biopsies were done at rest and 30 minutes after a single acute exercise bout in 14 healthy participants. Changes in mRNA expression were assessed using microarrays, and miRNA analysis was performed in a subset of 6 of the participants using sequencing techniques. Following exercise, 215 mRNAs were changed at the probe level (Bonferroni-corrected P<0.00000115). Pathway and Gene Ontology analysis showed enrichment in MAP kinase signaling, transcriptional regulation and DNA binding. Changes in several transcription factor mRNAs were correlated with insulin sensitivity, including MYC, r=0.71; SNF1LK, r=0.69; and ATF3, r= 0.61 (5 corrected for false discovery rate). Enrichment in the 5'-UTRs of exercise-responsive genes suggested regulation by common transcription factors, especially EGR1. miRNA species of interest that changed after exercise included miR-378, which is located in an intron of the PPARGC1B gene. CONCLUSIONS: These results indicate that transcription factor gene expression responses to exercise depend highly on insulin sensitivity in healthy people. The overall pattern suggests a coordinated cycle by which exercise and insulin sensitivity regulate gene expression in muscle.

Monovision laser in situ keratomileusis for pre-presbyopic and presbyopic patients.

PURPOSE: To evaluate the outcome and success of refractive surgery in pre-presbyopic and presbyopic patients with monovision after laser in situ keratomileusis (LASIK). METHODS: We performed a retrospective analysis of 748 eyes of 374 patients over 35 years of age treated in one or both eyes for a monovision outcome using the Alcon LADARVision 4000 laser. Analysis of age at the time of surgery, gender, bilateral versus unilateral treatment, targeted outcome, and patient acceptance are reported. RESULTS: The success rate of monovision patients in this study was 92.5%. Acceptance rate was higher in older patients, although there was no significantly greater acceptance of monovision for any of the variables studied (P values >.26). CONCLUSIONS: Monovision is a valuable option for pre-presbyopic and presbyopic patients considering refractive surgery.

Ferrara intrastromal corneal ring segments for severe keratoconus.

PURPOSE: Intrastromal Ferrara ring segments were inserted in eyes with severe keratoconus to evaluate safety and efficacy of this procedure. METHODS: Intrastromal Ferrara ring segments were placed in 36 eyes of 35 patients with severe keratoconus, who were evaluated after 12 months of follow-up. All patients had highly disabling visual acuity, contact lens intolerance, and a previous indication for penetrating keratoplasty. Statistical analysis included preoperative and postoperative uncorrected visual acuity, best spectacle-corrected visual acuity, manifest refraction, anterior chamber depth, keratometry, and pachymetry. RESULTS: No patient had a loss of visual acuity. Uncorrected visual acuity improved in 28 eyes (77.78%) and best spectacle-corrected visual acuity improved in 29 eyes (80.56%). Spherical equivalent refraction decreased from -7.29 +/- 3.12 D to -4.80 +/- 3.04 D at 12 months after Ferrara intrastromal ring segment implantation. Corneal topography and ultrasound biomicroscopy showed corneal flattening, demonstrated by thinning of the central cornea and a reduction in anterior chamber depth. Segment decentration occurred in one eye (2.7%), asymmetric positioning of the segments in two eyes (5%), inadequate depth in two eyes (5%), migration of the segments in two eyes (5%), segment extrusion in five eyes (13.8%), conjunctivitis in one eye (2.7%), bacterial keratitis in one eye (2.7%), and hydrops in one eye (2.7%). CONCLUSIONS: With these early results, Ferrara intrastromal ring segments appear to be an alternative for the treatment of severe keratoconus.

Increases in insulin sensitivity among obese youth are associated with gene expression changes in whole blood.

OBJECTIVE: Lifestyle intervention can improve insulin sensitivity in obese youth, yet few studies have examined the molecular signatures associated with these improvements. Therefore, the purpose of this study was to explore gene expression changes in whole blood that are associated with intervention-induced improvements in insulin sensitivity. METHODS: Fifteen (7M/8F) overweight/obese (BMI percentile = 96.3 ± 1.1) Latino adolescents (15.0 ± 0.9 years) completed a 12-week lifestyle intervention that included weekly nutrition education and 180 minutes of moderate-vigorous exercise per week. Insulin sensitivity was estimated by an oral glucose tolerance test and the Matsuda Index. Global microarray analysis profiling from whole blood was performed to examine changes in gene expression and to explore biological pathways that were significantly changed in response to the intervention. RESULTS: A total of 1,459 probes corresponding to mRNA transcripts (717 up, 742 down) were differentially expressed with a fold change ≥1.2. These genes were mapped within eight significant pathways identified, including insulin signaling, type 1 diabetes, and glycerophospholipid metabolism. Participants with increased insulin sensitivity exhibited five times the number of significant genes altered compared with nonresponders (1,144 vs. 230). CONCLUSIONS: These findings suggest that molecular signatures from whole blood are associated with lifestyle-induced health improvements among high-risk Latino youth.

Avaliação da contaminação por cobre e chumbo do lago Aratimbó - Umuarama - Pr / Evaluation of copper and lead contamination of the Aratimbó lake in Umuarama - Paraná

Este trabalho teve como objetivo quantificar os metais Cobre e Chumbo presentes na água e no sedimento do lago Aratimbó, localizado na cidade de Umuarama-PR e analisar os indicadores abióticos de temperatura e pH da água. As coletas foram feitas em triplicatas, em três pontos distintos, em dois diferentes dias, sendo o primeiro em tempo seco, anterior a chuva e outro logo após a primeira chuva. As amostras tiveram sua temperatura e pH verificado in loco e a determinação dos metais ocorreu por meio de Espectrofotometria de Absorção Atômica por chama. Os resultados de pH obtiveram valores satisfatórios em todos os pontos. Em relação à concentração de metais, na água, houve maior concentração de chumbo antes da chuva e uma concentração abaixo do limite apenas para o cobre no ponto 1. No sedimento, a concentração de ambos metais foi maior antes da chuva, exceto no ponto três, onde a concentração foi maior após a chuva. É possível que os maiores valores de concentração para os metais tenham sido superiores no período de seca em virtude do volume aumentado de água após a chuva, com maior diluição. Assim, podendo atribuir a contaminação às atividades antropogênicas, uma vez que o lago se localiza em região urbana central.

The purpose of this paper was to quantify the copper and lead present in the water and sediments of the Aratimbó lake, located in the city of Umuarama, in the state of Paraná, and to analyze the abiotic indicators of temperature and pH of the water. The samples were taken in triplicate at three different points, on two different days - the first during dry weather, before the rain, and the other after the first rain. The temperature and pH of the samples were verified in locu and the metals were determined through Atomic Absorption Spectrophotometry by flame. The pH results obtained satisfactory values ​​at all points. Regarding the concentration of metals in the water, a higher concentration of lead was observed before the rain and a concentration below the limit was observed only for copper in point 1. In the sediment, the concentration of both metals was higher before the rain, except in point three, where the concentration was also higher after the rain. It is possible that the highest concentration values ​​for the metals were higher in the dry period due to the increased volume of water after rainfall, with higher dilution of the metals, as well as the attribution of contamination to the anthropogenic activities, since the lake is located in a central urban region.

Association of Merkel cell polyomavirus-specific antibodies with Merkel cell carcinoma.

BACKGROUND: Merkel cell polyomavirus (MCPyV) has been detected in approximately 75% of patients with the rare skin cancer Merkel cell carcinoma. We investigated the prevalence of antibodies against MCPyV in the general population and the association between these antibodies and Merkel cell carcinoma. METHODS: Multiplex antibody-binding assays were used to assess levels of antibodies against polyomaviruses in plasma. MCPyV VP1 antibody levels were determined in plasma from 41 patients with Merkel cell carcinoma and 76 matched control subjects. MCPyV DNA was detected in tumor tissue specimens by quantitative polymerase chain reaction. Seroprevalence of polyomavirus-specific antibodies was determined in 451 control subjects. MCPyV strain-specific antibody recognition was investigated by replacing coding sequences from MCPyV strain 350 with those from MCPyV strain w162. RESULTS: We found that 36 (88%) of 41 patients with Merkel cell carcinoma carried antibodies against VP1 from MCPyV w162 compared with 40 (53%) of the 76 control subjects (odds ratio adjusted for age and sex = 6.6, 95% confidence interval [CI] = 2.3 to 18.8). MCPyV DNA was detectable in 24 (77%) of the 31 Merkel cell carcinoma tumors available, with 22 (92%) of these 24 patients also carrying antibodies against MCPyV. Among 451 control subjects from the general population, prevalence of antibodies against human polyomaviruses was 92% (95% CI = 89% to 94%) for BK virus, 45% (95% CI = 40% to 50%) for JC virus, 98% (95% CI = 96% to 99%) for WU polyomavirus, 90% (95% CI = 87% to 93%) for KI polyomavirus, and 59% (95% CI = 55% to 64%) for MCPyV. Few case patients had reactivity against MCPyV strain 350; however, indistinguishable reactivities were found with VP1 from strain 350 carrying a double mutation (residues 288 and 316) and VP1 from strain w162. CONCLUSION: Infection with MCPyV is common in the general population. MCPyV, but not other human polyomaviruses, appears to be associated with Merkel cell carcinoma.

ATG-anchored AFLP (ATG-AFLP) analysis in cotton.

Amplified fragment length polymorphism (AFLP) marker system has had broad applications in biology. However, the anonymous AFLP markers are mainly amplified from non-coding regions, limiting their usefulness as a functional marker system. To take advantages of the traditional AFLP techniques, we propose substitution of a restriction enzyme that recognizes a restriction site containing ATG, called ATG-anchored AFLP (ATG-AFLP) analysis. In this study, we chose NsiI (recognizing ATGCAT) to replace EcoRI in combination with MseI to completely digest genomic DNA. One specific adaptor, one pre-selective primer and six selective amplification primers for the NsiI site were designed for ligation and PCR. Six NsiI and eight MseI primers generated a total of 1,780 ATG-AFLP fragments, of which 750 (42%) were polymorphic among four genotypes from two cultivated cotton species (Upland cotton, Gossypium hirsutum and Pima cotton, G. barbadense). The number of ATG-AFLP markers was sufficient to separate the four genotypes into two groups, consistent with their evolutionary and breeding history. Our results also showed that ATG-AFLP generated less number of total and polymorphic fragments per primer combination (2-3 vs. 4-5) than conventional AFLP within Upland cotton. Using a recombination inbred line (RIL) population, 62 polymorphic ATG-AFLP markers were mapped to 19 linkage groups with known chromosome anchored simple sequence repeat (SSR) markers. Of the nine ATG-AFLP fragments randomly chosen, three were found to be highly homologous to cotton cDNA sequences. An in-silico analysis of cotton and Arabidopsis cDNA confirmed that the ATG-anchored enzyme combination NsiI/MseI did generate more fragments than the EcoRI/MseI combination.

Vaginal misoprostol prior to diagnostic hysteroscopy in patients of reproductive age: a randomized clinical trial.

STUDY OBJECTIVE: To assess pain during and after diagnostic hysteroscopy in patients of reproductive age according to use of vaginal misoprostol. DESIGN: A randomized, triple-masked, controlled clinical trial (Canadian Task Force classification I). SETTING: Diagnostic center at Instituto Materno Infantil de Pernambuco-Brazil. PATIENTS: A total of 45 women of reproductive age underwent diagnostic anesthesia-free hysteroscopy. INTERVENTIONS: In all, 400 mg of vaginal misoprostol or placebo was administered to randomized patients before diagnostic hysteroscopy. MEASUREMENTS AND MAIN RESULTS: The clinical trial was interrupted in patients receiving misoprostol because of significant vaginal bleeding precluding the procedure (p = .0006). No significant difference existed in pain scores between the groups. Vaginal bleeding was the main side effect occurring in 11 patients of the misoprostol group. No vaginal bleeding occurred in the placebo group (p = .00002). CONCLUSION: In the doses used, vaginal misoprostol induced vaginal bleeding and precluded diagnostic hysteroscopy in patients of reproductive age that limits continuation of this line of research. Further studies are needed to reassess the use of the drug in patients of this age group.

Comparison of flap thickness reproducibility using microkeratomes with a second motor for advancement.

PURPOSE: To compare flap thickness and reproducibility of four different types of microkeratomes during LASIK. DESIGN: Retrospective, nonrandomized, comparative case series. PARTICIPANTS: Four hundred ninety consecutive eyes underwent LASIK and were evaluated by measuring the central flap thickness by subtractive pachymetry. METHODS: All flaps were created using the Bausch & Lomb (Miami, FL) Hansatome 180 head, the Alcon (Fort Worth, TX) Summit Krumeich Barraquer Microkeratome 160 head, the Moria (Antony, France) Carriazo Barraquer (CB) 130 head, or the Moria M2 110 head. RESULTS: The flap thickness measurements differed according to the microkeratome used and were 131+/-28 microm in 41 eyes (8.4%) with the Bausch & Lomb Hansatome 180 head, 162+/-21 microm in 127 eyes (25.9%) with the Alcon Summit Krumeich Barraquer Microkeratome (SKBM) 160 head, 157+/-40 microm in 65 eyes (13.3%) with the Moria CB 130 head, and 134+/-23 microm in 257 eyes (52.4%) with the Moria M2 110 head. The central flap thickness with the SKBM and Moria M2 was statistically significantly more reproducible than with the Moria CB (P< 0.0005). There is no correlation between flap thickness reproducibility and age, corneal thickness, or corneal keratometric values. However, considering all the microkeratomes, female gender had statistically significantly more variability than male gender (P<0.02). CONCLUSIONS: Based on these results, the greatest predictability of flap thickness was seen with the SKBM and Moria M2 microkeratomes, which both use a second motor for advancement. The greatest variability, noted with the Moria CB, was likely due to the manual translation feature and places further importance on the safety of the second motor and automation when performing LASIK.