Biotypes and serotypes of Haemophilus influenzae ocular isolates.
To determine which subtypes of Haemophilus influenzae are most commonly associated with ocular disease, and whether the site of ocular H influenzae infection is correlated with specific subtypes of the organism. The biotypes and serotypes of ocular H influenzae isolates collected at the Francis I Proctor Foundation between March 1989 and January 2000 were examined. A total of 62 ocular isolates were retrieved from frozen storage and plated on chocolate agar. Biotypes were assigned based upon the ability of the isolates to produce indole, urease, and ornithine decarboxylase. Capsular subtypes a-f were determined by slide agglutination using commercially available subtype specific antisera. Identified biotypes and serotypes were then analysed with regard to site of infection. Patient age ranged from 1 to 92 years with a median age of 45 years. 38 (61%) of the isolates were biotype II, 23 (37%) were biotype III, and one (2%) was biotype VII. All of the isolates were non-encapsulated and thus serologically non-typable. H influenzae biotype II was found in 28 of 48 (58%) conjunctivitis cases, five of eight (63%) keratitis cases, and two of two (100%) endophthalmitis cases. Biotype III was found in 20 of 48 (42%) conjunctivitis cases, two of eight (25%) keratitis cases, and a single case of dacryocystitis. Biotype VII was associated with one of eight (13%) keratitis cases. Most ocular H influenzae isolates appear to be serologically non-typable strains from biotypes II and III, less virulent subtypes that frequently colonise the nasopharynx. In addition, the site of ocular H influenzae infections appears to be largely independent of species subtype.


Xuezhikang, an extract of cholestin, decreases plasma inflammatory markers and endothelin-1, improve exercise-induced ischemia and subjective feelings in patients with cardiac syndrome X.
Previous studies have demonstrated that Xuezhikang, an extract of cholestin, available from Chinese red yeast rice, could effectively modify lipid profile. The present study was undertaken to investigate whether Xuezhikang could modify endothelin-1 (ET-1), interleukin-6 (IL-6), high-sensitivity C-reactive protein (CRP) and exercise-induced ischemia in patients with cardiac syndrome X (CSX). Thirty-six patients with CSX were randomly assigned to 1200 mg/d of Xuezhikang or placebo group (n=18 respectively). Blood samples were drawn at day 0 and day 90 for measuring above parameters. The treadmill exercise tests and subjective feelings were also assessed at day 0 and day 90. The data showed that Xuezhikang therapy resulted in significant reductions in total cholesterol (TC, 19%), low-density lipoprotein cholesterol (LDL-C) (26%), and triglycerides (TG) compared with baseline (16%, p<0.01 respectively). The data also showed that Xuezhikang led significantly to reductions in median and log-CRP levels (38% and 44%, p<0.01 respectively), IL-6 (20%, p<0.01), and ET-1 (47%, p<0.01) compared with baseline. The exercise duration, and time to 1 mm ST-segment depression was significantly prolonged after Xuezhikang therapy (9% and 6%, p<0.05 respectively) accompanied by improvement of subjective feelings. Data suggested that the benefit of Xuezhikang resulted in significant modification vascular function by reduction of ET-1, inflammatory markers and LDL cholesterol, which may be clinically important for patients with CSX.


Faculty Support for Self-Directed Learning in Internal Medicine Residency: A Qualitative Study Using Grounded Theory.
Self-directed learning (SDL) is part of residency training, which residents desire guidance in implementing. To characterize SDL within the clinical context, this study explored residents' perceptions of faculty members' role in promoting and supporting resident SDL. Using a constructivist grounded theory approach, the authors conducted seven focus groups with 46 internal medicine residents at the Mayo Clinic Internal Medicine Residency Program from October 2014 to January 2015. Focus group transcripts were deidentified and processed through open coding and analytic memo writing. Guided by a previously developed SDL model, data were analyzed regarding faculty member involvement in resident SDL. Themes were organized and patterns were discussed at team meetings, with constant comparison with new data. Trustworthiness was established using two member-check sessions. The authors identified themes within the categories of faculty guidance for SDL, SDL versus other-directed learning (ODL), and faculty archetypes for supporting SDL. Clinical teachers play a key role in facilitating resident SDL and can provide guidance at each step in the SDL process. Residents discussed the distinction between SDL and ODL, highlighting the integrated nature of learning and interplay between the two approaches to learning. Residents identified themes relating to three archetypal approaches faculty implement to support resident SDL in the clinical environment (directed, collaborative, and role model SDL), with benefits and challenges of each approach. This study underscores the importance of external guidance for resident SDL and expands on approaches faculty members can use to support SDL in the clinical context.


Enhancing the delivery of cancer care to the disadvantaged: the challenge to providers.
Few attempts have been made to document and highlight the challenges faced by professionals who work with medically and economically disadvantaged persons seeking cancer care. This report analyzes and summarizes the testimony presented by such professionals at a series of nationwide hearings in 1989 sponsored by the American Cancer Society. The purpose of the hearings was to document the effects of economic and medical disadvantage on the development of cancer and cancer care. Among those presenting testimony were persons with personal cancer experiences, family members, and partners of cancer patients, healthcare providers, and related community liaisons. Professionals who worked with disadvantaged persons shared their experiences during the hearings. They suggested that the beliefs and attitudes of their peers were a major deterrent to providing cancer care. Processes and systems discouraged referrals. Lack of access to resources to serve the needs of the disadvantaged was the greatest obstacle to care.


High incidence of ICA anterior wall aneurysms in patients with an anomalous origin of the ophthalmic artery: possible relevance to the pathogenesis of aneurysm formation.
Surgery for aneurysms at the anterior wall of the internal carotid artery (ICA), which are also referred to as ICA anterior wall aneurysms, is often challenging. A treatment strategy needs to be determined according to the pathology of the aneurysm-namely, whether the aneurysm is a saccular aneurysm with firm neck walls that would tolerate clipping or coiling, a dissecting aneurysm, or a blood blister-like aneurysm. However, it is not always possible to properly evaluate the condition of the aneurysm before surgery solely based on angiographic findings. The authors focused on the location of the ophthalmic artery (OA) in determining the pathology of ICA anterior wall aneurysms. Between January 2006 and December 2012, diagnostic cerebral angiography, for any reason, was performed on 1643 ICAs in 855 patients at Saitama Medical Center. The authors also investigated the relationship between the origin of the OA and the incidence of ICA anterior wall aneurysms. The pathogenesis was also evaluated for each aneurysm based on findings from both angiography and open surgery to identify any correlation between the location where the OA originated and the conditions of the aneurysm walls. Among 1643 ICAs, 31 arteries (1.89%) were accompanied by an anomalous origin of the OA, including 26 OAs originating from the C3 portion, 3 originating from the C4 portion, and 2 originating from the anterior cerebral artery. The incidence of an anomalous origin of the OA had no relationship to age, sex, or side. Internal carotid artery anterior wall aneurysms were observed in 16 (0.97%) of 1643 ICAs. Female patients had a significantly higher risk of having ICA anterior wall aneurysms (p = 0.026). The risk of ICA anterior wall aneurysm formation was approximately 50 times higher in patients with an anomalous origin of the OA (25.8% [8 of 31]) than in those with a normal OA (0.5% [8 of 1612], p < 0.0001). Based on angiographic classifications, saccular aneurysms were significantly more common in patients with an anomalous origin of the OA than in those with a normal OA (p = 0.041). Ten of 16 patients with ICA anterior wall aneurysms underwent craniotomies. Based on the intraoperative findings, all 6 aneurysms with normal OAs were dissecting or blood blister-like aneurysms, not saccular aneurysms. There was a close relationship between the location of the OA origin and the predisposition to ICA anterior wall aneurysms. Developmental failure of the OA and subsequent weakness of the vessel wall might account for this phenomenon, as previously reported regarding other aneurysms related to the anomalous development of parent arteries. The data also appear to indicate that ICA anterior wall aneurysms in patients with an anomalous origin of the OA tend to be saccular aneurysms with normal neck walls. These findings provide critical information in determining therapeutic strategies for ICA anterior wall aneurysms.


Mitochondrial donation, patient engagement and narratives of hope.
This article develops the sociology of hope and patient engagement by exploring how patients' perceptions and actions are shaped by narratives of hope surrounding the clinical introduction of novel reproductive techniques. In 2015, after extensive public debates, the UK became the first country to legalise a mitochondrial donation technique aimed at preventing the transmission of inherited disorders. The article draws on the accounts of twenty-two women of reproductive age who are at risk of having a child with mitochondrial disease and would be the potential target of the technique. We explore the extent to which our participants engaged with the public debates and how they accounted for their support of mitochondrial donation. We show that while the majority of our participants were in favour of legalisation, they did not necessarily wish to use the technique themselves. We found that hope was multi-faceted, involving hope for self, hope for family and hope for society. We conclude by considering the implications of hope narratives for patients and families and the important but potentially limited role that patients can play as advocates for technology.


Flexibility and resilience of great tit (Parus major) gut microbiomes to changing diets.
Gut microbial communities play important roles in nutrient management and can change in response to host diets. The extent of this flexibility and the concomitant resilience is largely unknown in wild animals. To untangle the dynamics of avian-gut microbiome symbiosis associated with diet changes, we exposed Parus major (Great tits) fed with a standard diet (seeds and mealworms) to either a mixed (seeds, mealworms and fruits), a seed, or a mealworm diet for 4 weeks, and examined the flexibility of gut microbiomes to these compositionally different diets. To assess microbiome resilience (recovery potential), all individuals were subsequently reversed to a standard diet for another 4 weeks. Cloacal microbiomes were collected weekly and characterised through sequencing the v4 region of the 16S rRNA gene using Illumina MiSeq. Initial microbiomes changed significantly with the diet manipulation, but the communities did not differ significantly between the three diet groups (mixed, seed and mealworm), despite multiple diet-specific changes in certain bacterial genera. Reverting birds to the standard diet led only to a partial recovery in gut community compositions. The majority of the bacterial taxa that increased significantly during diet manipulation decreased in relative abundance after reversion to the standard diet; however, bacterial taxa that decreased during the manipulation rarely increased after diet reversal CONCLUSIONS: The gut microbial response and partial resilience to dietary changes support that gut bacterial communities of P. major play a role in accommodating dietary changes experienced by wild avian hosts. This may be a contributing factor to the relaxed association between microbiome composition and the bird phylogeny. Our findings further imply that interpretations of wild bird gut microbiome analyses from single-time point sampling, especially for omnivorous species or species with seasonally changing diets, should be done with caution. The partial community recovery implies that ecologically relevant diet changes (e.g., seasonality and migration) open up gut niches that may be filled by previously abundant microbes or replaced by different symbiont lineages, which has important implications for the integrity and specificity of long-term avian-symbiont associations.


APOBEC3G governs the generation of truncated AATF protein to ensure oncogenic transformation.
The oncogenic potential of Apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3G (APOBEC3G) was recently appreciated by the finding that revealed its ability to downregulate Kruppel-like factor 4 (KLF4) gene translation through its affinity for 3'UTR of KLF4 mRNA. Keeping in view the fact that KLF4 is known to repress apoptosis antagonizing transcription factor (AATF) gene expression, the present study employed stem cells as archetype model to explore the effect of APOBEC3G over-expression upon AATF gene expression within these cells as well as on the genes involved in oncogenic transformation. Such a study revealed that APOBEC3G had the ability to bind AATF mRNA within its third exon to facilitate the generation of truncated 23 kDa AATF translation product which, in turn, had the inherent capacity to be the crucial mediator of APOBEC3G induced oncogenic transformation within such cells.


In-Vehicle Alcohol Detection Using Low-Cost Sensors and Genetic Algorithms to Aid in the Drinking and Driving Detection.
Worldwide, motor vehicle accidents are one of the leading causes of death, with alcohol-related accidents playing a significant role, particularly in child death. Aiming to aid in the prevention of this type of accidents, a novel non-invasive method capable of detecting the presence of alcohol inside a motor vehicle is presented. The proposed methodology uses a series of low-cost alcohol MQ3 sensors located inside the vehicle, whose signals are stored, standardized, time-adjusted, and transformed into 5 s window samples. Statistical features are extracted from each sample and a feature selection strategy is carried out using a genetic algorithm, and a forward selection and backwards elimination methodology. The four features derived from this process were used to construct an SVM classification model that detects presence of alcohol. The experiments yielded 7200 samples, 80% of which were used to train the model. The rest were used to evaluate the performance of the model, which obtained an area under the ROC curve of 0.98 and a sensitivity of 0.979. These results suggest that the proposed methodology can be used to detect the presence of alcohol and enforce prevention actions.


Anal incontinence and unrecognized anal sphincter injuries after vaginal delivery- a cross-sectional study in Norway.
Aim of the study was to estimate the prevalence of postpartum anal incontinence among women who delivered vaginally, and to assess the extent to which obstetric injuries to the anal sphincters are missed. All women (both primiparous and multiparous) who delivered vaginally and received any kind of sutures in the perineal area at Innlandet Hospital Trust Elverum in Norway between January 1, 2015 and June 30, 2016 were invited to answer a questionnaire on St. Mark's incontinence score and to participate in a clinical examination of the pelvic floor including endoanal sonography. In total 52,3% (n = 207) of the 396 invited women participated in the study. Mean St. Mark's score was 1.8 points (95% CI 1.4 to 2.1) at examination 14 months (mean) postpartum, and none of the participants suffered from weekly fecal leakage. Fecal urgency affected 11.7% (95% CI 7.1 to 16.3) of the participants, and 8.7% (95%CI 5.1 to 12.8) had weekly involuntary leakage of flatus. Nine women (9.3%, 95% CI 4.1 to 15.5) had a previously undetected third degree obstetric anal sphincter injury. The prevalence of anal incontinence among women who have delivered vaginally and received sutures due to 1st and 2nd degree perineal lacerations is low. Some obstetric anal sphincter injuries remain unrecognized at the time of delivery, but the symptoms of anal incontinence due to these injuries are in the lower half of the St. Mark's incontinence score. Women with persistent symptoms like fecal urgency or leakage of gas and/or feces should be referred to evaluation by a colorectal surgeon in order to achieve optimal treatment.


Overuse Injuries of the Pediatric and Adolescent Throwing Athlete.
In the hypercompetitive environment of year round youth baseball, arm pain is commonplace. Although much research has been done about injuries in the overhead throwing athlete, the emphasis has been on the more elite levels, where athletes have reached full development. The anatomy of the skeletally immature athlete, including open physeal plates and increased tissue laxity, raises unique issues in the presentation and treatment of repetitive throwing injuries of the elbow and shoulder. With a focus on "little leaguers," this discussion evaluates five of the most common elbow and shoulder injuries-Little Leaguer's elbow, ulnar collateral ligament sprain or tear, osteochondritis dissecans/Panner's disease, Little Leaguer's shoulder, and multidirectional instability. In the developmentally distinct pediatric athlete, pathogenesis, diagnosis, and treatment may differ from that established for adults. Offering early diagnosis and treatment appropriate to a child's level of development will enable youth to not only continue to play sports but to also maintain full functionality as active adults.


Genetic diversity of the Japanese wood pigeon, Columba janthina, endemic to islands of East Asia, estimated by newly developed microsatellite markers.
The Japanese wood pigeon Columba janthina is endemic to islands of East Asia and is listed as Near Threatened by the International Union for Conservation of Nature (IUCN). One subspecies, C. janthina nitens, in particular, is at the greatest risk of extinction due to its small population size. To reduce the extinction risk of C. janthina, it is important to understand the species' present genetic status and to develop an appropriate conservation plan based on genetic data. We developed seven new microsatellite markers for two subspecies of C. janthina: C. janthina janthina and C. janthina nitens. We also confirmed the cross-use of one microsatellite marker developed for Columba livia var. domestica. Seven loci were polymorphic in C. janthina janthina, while two loci were polymorphic in C. janthina nitens. Using the markers, we performed preliminary analysis of genetic diversity and genetic structure within each subspecies. The expected heterozygosity ranged from 0.00 to 0.64 in C. janthina janthina and from 0.00 to 0.08 in C. janthina nitens. Each subspecies and each population within C. janthina janthina had different allele frequencies. C. janthina nitens exhibited far lower genetic diversity than C. janthina janthina. Furthermore, C. janthina nitens appears to have experienced strong genetic drift from a common ancestral population, inferred by STRUCTURE analysis. The markers described here may be useful for investigating genetic diversity and genetic structure of C. janthina populations, and could be used to estimate appropriate evolutionary significant unit and to guide development of a captive breeding program based on genetic information.


Maternal mental health mediates the effects of pandemic-related stressors on adolescent psychopathology during COVID-19.
This study examined whether COVID-19-related maternal mental health changes contributed to changes in adolescent psychopathology. A community sample of 226 adolescents (12 years old before COVID-19) and their mothers were asked to complete COVID-19 surveys early in the pandemic (April-May 2020, adolescents 14 years) and approximately 6 months later (November 2020-January 2021). Surveys assessed pandemic-related stressors (health, financial, social, school, environment) and mental health. Lower pre-pandemic family income-to-needs ratio was associated with higher pre-pandemic maternal mental health symptoms (anxiety, depression) and adolescent internalizing and externalizing problems, and with experiencing more pandemic-related stressors. Pandemic-related stressors predicted increases in maternal mental health symptoms, but not adolescent symptoms when other variables were covaried. Higher maternal mental health symptoms predicted concurrent increases in adolescent internalizing and externalizing. Maternal mental health mediated the effects of pre-pandemic income and pandemic-related stressors on adolescent internalizing and externalizing problems. Results indicate that adolescent mental health is closely tied to maternal mental health during community-level stressors such as COVID-19, and that pre-existing family economic context and adolescent symptoms increase risk for elevations in symptoms of psychopathology.


Activation of electropermeabilized neutrophils by adenosine 5'-[gamma-thio]triphosphate (ATP[S]). Role of phosphatases in stimulus-response coupling.
Electrically permeabilized human neutrophils were used to study the mechanism of activation of the NADPH oxidase by chemotactic factors. The respiratory burst elicited by formyl-methionyl-leucyl-phenylalanine (fMLP) was strictly dependent on the addition of ATP. The response was also supported by adenosine 5'-[gamma-thio]triphosphate (ATP[S]), but not by the non-hydrolysable analogue (p[NH]ppA). In the presence of ATP, displacement of fMLP from its receptor by antagonist peptides resulted in the abrupt termination of the O2-consumption burst. In contrast, the response persisted after displacement of fMLP when ATP[S] was present. This finding is consistent with the formation of biologically active thiophosphoproteins which are resistant to cleavage by cellular phosphatases. Accordingly, lower concentrations of ATP[S], as compared with ATP, were required to support the fMLP response. The data indicate that protein phosphatases control the extent and duration of the response in cells stimulated with chemoattractants. Unlike ATP, sub-millimolar concentrations of ATP[S] elicited a spontaneous respiratory burst in the absence of fMLP or other stimuli. This effect was inhibited by p[NH]ppA and was not observed in intact (non-permeabilized) cells, indicating interaction of ATP[S] with an intracellular adenine-nucleotide-binding site, possibly a protein kinase. These results suggest that protein kinases are active in neutrophils in the absence of exogenous stimuli, but that accumulation of the essential phosphoprotein(s) is normally prevented by the ongoing vigorous phosphatase activity. It is conceivable that control of the respiratory burst is exerted by inhibition of phosphatase activity, instead of or in addition to the more commonly postulated activation of protein kinases.


Confronting and Coping with Multidrug-Resistant Tuberculosis: Life Experiences in Thailand.
In this article, we aimed to understand the life experiences of Thai persons diagnosed with multi-drug-resistant tuberculosis (MDR-TB). A qualitative study using a face-to-face in-depth interview was conducted at a hospital in Thailand which has the highest prevalence of MDR-TB in the country between January and February 2019. Twenty persons living with MDR-TB in Thailand were purposively selected to represent a variety of experiences based on different gender, ages, and treatment phases. Qualitative data were transcribed and thematic analysis was applied to identify common themes and sub-themes. The results indicated that all participants faced emotional difficulties, such as fear of death, fear of stigmatization, confusion, and sadness when first knowing of their diagnosis. Family and social support were the main ways that the patients coped with difficult situations. Suicidal ideas were more prevalent among patients with poor family support. Screening for mental health problems should be routinely performed in MDR-TB patients. Proper health education should be provided to patients and families to reduce emotional difficulties and stigmatization.


Effects of heating on composition, degree of darkness, and stacking nanostructure of soil humic acids.
Wildfires and prescribed burning can affect both the quality and the quantity of organic matter in soils. In this study, we investigated qualitative and quantitative changes of soil humic substances in two different soils (an Entisol from a paddy field and an Inceptisol from a cedar forest) under several controlled heating conditions. Soil samples were heated in a muffle furnace at 200, 250, or 300 °C for 1, 3, 5, or 12h. The humic acid and fulvic acid contents of the soil samples prior to and after heating were determined. The degree of darkness, elemental composition, carbon and nitrogen stable isotope ratios, (13)C nuclear magnetic resonance spectra, and X-ray diffraction patterns of humic acids extracted from the soils before and after heating were measured. The proportion of humic acids in total carbon decreased with increasing heating time at high temperature (300 °C), but increased with increasing heating time at ≤ 250 °C. The degree of darkness of the humic acids increased with increasing heating time and temperature. During darkening, the H/C atomic ratios, the proportion of aromatic C, and the carbon and nitrogen stable isotope ratios increased, whereas the proportions of alkyl C and O-alkyl C decreased. X-ray diffraction analysis verified that a stacking nanostructure developed by heating. Changes in the chemical structure of the humic acids from the heated soils depended on the type of soil. The major structural components of the humic acids from the heated Entisol were aromatic C and carboxylic C, whereas aliphatic C, aromatic C, and carboxylic C structural components were found in the humic acids from the heated Inceptisol. These results suggest that the heat-induced changes in the chemical structure of the humic acids depended on the source plant.


Sorafenib for the treatment of advanced renal cell carcinoma.
This report describes the U.S. Food and Drug Administration (FDA) review and approval of sorafenib (Nexavar, BAY43-9006), a new small-molecule, oral, multi-kinase inhibitor for the treatment of patients with advanced renal cell carcinoma (RCC). After meeting with sponsors during development studies of sorafenib, the FDA reviewed the phase 3 protocol under the Special Protocol Assessment mechanism. Following new drug application submission, FDA independently analyzed the results of two studies in advanced RCC: a large, randomized, double-blinded, phase 3 international trial of single-agent sorafenib and a supportive phase 2 study. In the phase 3 trial, 902 patients with advanced progressive RCC after one prior systemic therapy were randomized to 400 mg sorafenib twice daily plus best supportive care or to a matching placebo plus best supportive care. Primary study end points included overall survival and progression-free survival (PFS). A PFS analysis, pre-specified and conducted after a total of 342 events, showed statistically significant superiority for the sorafenib group (median = 167 days) compared with that for the controls (median = 84 days, log-rank P < 0.000001); the sorafenib/placebo hazard ratio was 0.44 (95% confidence interval, 0.35-0.55). Results were similar regardless of patient risk score, performance status, age, or prior therapy. The (partial) response rate to sorafenib was 2.1%. Overall survival results are preliminary. The principal toxicities in the sorafenib patients included reversible skin rashes in 40% and hand-foot skin reaction in 30%; diarrhea was reported in 43%, treatment-emergent hypertension was reported in 17%, and sensory neuropathic changes were reported in 13%. Grade 4 adverse events were uncommon. Grade 3 adverse events were hand-foot skin reaction (6%), fatigue (5%), and hypertension (3%). Laboratory findings included asymptomatic hypophosphatemia in 45% of sorafenib patients versus 11% in the placebo arm and elevation of serum lipase in 41% of sorafenib patients versus 30% in the placebo arm. Grade 4 pancreatitis was reported in two sorafenib patients, although both patients subsequently resumed sorafenib, with one at full dose. Sorafenib received FDA regular approval on December 20, 2005 for the treatment of advanced RCC based on the persuasive magnitude of improvement in PFS with acceptable safety. The recommended dose is 400 mg (two 200-mg tablets) twice daily taken either 1 h before or 2 h after meals. Adverse events were accommodated by temporary dose interruptions or reductions.


Bayesian biclustering for microbial metagenomic sequencing data via multinomial matrix factorization.
High-throughput sequencing technology provides unprecedented opportunities to quantitatively explore human gut microbiome and its relation to diseases. Microbiome data are compositional, sparse, noisy, and heterogeneous, which pose serious challenges for statistical modeling. We propose an identifiable Bayesian multinomial matrix factorization model to infer overlapping clusters on both microbes and hosts. The proposed method represents the observed over-dispersed zero-inflated count matrix as Dirichlet-multinomial mixtures on which latent cluster structures are built hierarchically. Under the Bayesian framework, the number of clusters is automatically determined and available information from a taxonomic rank tree of microbes is naturally incorporated, which greatly improves the interpretability of our findings. We demonstrate the utility of the proposed approach by comparing to alternative methods in simulations. An application to a human gut microbiome data set involving patients with inflammatory bowel disease reveals interesting clusters, which contain bacteria families Bacteroidaceae, Bifidobacteriaceae, Enterobacteriaceae, Fusobacteriaceae, Lachnospiraceae, Ruminococcaceae, Pasteurellaceae, and Porphyromonadaceae that are known to be related to the inflammatory bowel disease and its subtypes according to biological literature. Our findings can help generate potential hypotheses for future investigation of the heterogeneity of the human gut microbiome.


Small and large B cells respond differently to T cell-derived B cell growth and differentiation factors.
A major problem in B cell biology is the determination of the roles played by helper T (TH) cells vs cytokines in the activation, replication, and differentiation of B lymphocytes. There is general agreement that activated B cells in cycle can replicate and terminally differentiate when provided with appropriate T cell-derived lymphokines. There is considerable controversy, however, as to whether cytokines can induce resting G0 B cells to secrete IgM. Some reports claim that TH cells are required before cytokines can act. In contrast, other reports claim that some T cell-derived supernatants (SN) have the capacity to activate resting B cells to become immunoglobulin-secreting cells. In the present study, we have examined one such SN (S26.5) as well as the EL-4 and PK 7.1 SN for their capacity to activate resting G0 B cells and a population of less dense B cells containing activated cells. These two populations, separated by Percoll density centrifugation, were characterized for size, stage in the cell cycle, and cell surface phenotype. It was shown that the most dense population contained predominantly G0 B cells, whereas the less dense population contained a subset of cells in cycle. Our studies show that neither T cell SN nor, indeed, any combination of cytokines and anti-immunoglobulins caused a major increase in the number of cells secreting IgM in the population enriched in G0 cells. In contrast, the T cell SN caused marked increases in the generation of IgM secreting cells in the population that contained a large proportion of activated cells. Limiting dilution analysis confirmed that the number of responding cells in the dense cell population was substantially lower than the number of responding cells in the less dense population. The small number of precursors in the dense B cell population may be attributed to contamination of that population with cells that have undergone activation steps in vivo. The present results, therefore, add further evidence to an existing large body of evidence that TH cells are essential for the terminal differentiation of G0 resting B cells in response to thymus-dependent antigens.


Effects of eicosapentaenoic and docosahexaenoic n-3 fatty acids from fish oil and preferential Cox-2 inhibition on systemic syndromes in patients with advanced lung cancer.
Under the common denomination of Systemic Immune-Metabolic Syndrome (SIMS), we grouped many symptoms that share a similar pathophysiologic background. SIMS is the result of the dysfunctional interaction of tumor cells, stroma cells, and the immune system, leading to the release of cytokines and other systemic mediators such as eicosanoids. SIMS includes systemic syndromes such as paraneoplastic hemopathies, hypercalcemia, coagulopathies, fatigue, weakness, cachexia, chronic nausea, anorexia, and early satiety among others. Eicosapentaenoic and docosahexaenoic n-3 fatty acids from fish oil can help in the management of persistent chronic inflammatory states, but treatment's compliance is generally poor. Preferentially, Cox-2 inhibition can create a favorable pattern of cytokines by decreasing the production of certain eicosanoids, although their role in SIMS is unknown. The aim of this study was to test the hypothesis that by modulating systemic inflammation through an eicosanoid-targeted approach, some of the symptoms of the SIMS could be controlled. We exclusively evaluated 12 patients for compliance. Patients were assigned 1 of the 4 treatment groups (15-, 12-, 9-, or 6-g dose, fractionated every 8 h). For patients assigned to 15 and 12 doses, the overall compliance was very poor and unsatisfactory for patients receiving the 9-g dose. The maximum tolerable dose was calculated to be around 2 capsules tid (6 g of fish oil per day). A second cohort of 22 patients with advanced lung cancer and SIMS were randomly assigned to receive either fish oil, 2 g tid, plus placebo capsules bid (n = 12) or fish oil, 2 g tid, plus celecoxib 200 mg bid (n = 10). All patients in both groups received oral food supplementation. After 6 wk of treatment, patients receiving fish oil + placebo or fish oil + celecoxib showed significantly more appetite, less fatigue, and lower C-reactive protein (C-RP) values than their respective baselines values (P < 0.02 for all the comparisons). Additionally, patients in the fish oil + celecoxib group also improved their body weight and muscle strength compared to baseline values (P < 0.02 for all the comparisons). Comparing both groups, patients receiving fish oil + celecoxib showed significantly lower C-RP levels (P = 0.005, t-test), higher muscle strength (P = 0.002, t-test) and body weight (P = 0.05, t-test) than patients receiving fish oil + placebo. The addition of celecoxib improved the control of the acute phase protein response, total body weight, and muscle strength. Additionally, the consistent nutritional support used in our patients could have helped to maximize the pharmacological effects of fish oil and/or celecoxib. This study shows that by modulating the eicosanoid metabolism using a combination of n-3 fatty acids and cyclooxygenase-2 inhibitor, some of the signs and symptoms associated with a SIMS could be ameliorated.


Hydrostatic pressure and the actomyosin cortex drive mitotic cell rounding.
During mitosis, adherent animal cells undergo a drastic shape change, from essentially flat to round. Mitotic cell rounding is thought to facilitate organization within the mitotic cell and be necessary for the geometric requirements of division. However, the forces that drive this shape change remain poorly understood in the presence of external impediments, such as a tissue environment. Here we use cantilevers to track cell rounding force and volume. We show that cells have an outward rounding force, which increases as cells enter mitosis. We find that this mitotic rounding force depends both on the actomyosin cytoskeleton and the cells' ability to regulate osmolarity. The rounding force itself is generated by an osmotic pressure. However, the actomyosin cortex is required to maintain this rounding force against external impediments. Instantaneous disruption of the actomyosin cortex leads to volume increase, and stimulation of actomyosin contraction leads to volume decrease. These results show that in cells, osmotic pressure is balanced by inwardly directed actomyosin cortex contraction. Thus, by locally modulating actomyosin-cortex-dependent surface tension and globally regulating osmotic pressure, cells can control their volume, shape and mechanical properties.


Phytochrome A Mediates the Disassembly of Processing Bodies in Far-Red Light.
Phytochromes are red- and far-red light receptors that control the growth and development of plants, enabling them to respond adequately to changing light conditions. It has been shown that halted mRNAs stored in RNA granules called processing bodies are released upon light perception and contribute to the adaptation to the light environment. However, the photophysiological background of this process is largely unknown. We found that light of different wavelengths can trigger the disassembly of processing bodies in a dose- and time-dependent manner. We show that phytochromes control this process in red- and far-red light and that cytoplasmic phytochrome A is sufficient and necessary for the far-red light-induced disassembly of processing bodies. This adds a novel, unexpected cytoplasmic function to the processes controlled by phytochrome A. Overall, our findings suggest a role of phytochromes in the control of translationally halted mRNAs that are stored in processing bodies. We expect our findings to facilitate understanding of how light and environmental cues control the assembly and disassembly of processing bodies, which could have broader implications for the regulation of non-membranous organelles in general.


Profile of Kawasaki disease at a tertiary care center in India.
Kawasaki disease (KD) is the most common cause of acquired heart disease in developed countries. KD is increasingly being reported from India; however, studies involving the large number of patients are few. All children presenting to the center from January 2017 to December 2019, diagnosed to have KD, were retrospectively included in the study. Clinical and laboratory profiles, including echocardiograms, were reviewed. Factors contributing to intravenous immunoglobulin (IVIg) refractoriness and the development of coronary artery abnormalities (CAA) were assessed. A total of 39 children with KD presented to the center during the study. While 32 received initial treatment at our center, seven were referred after the initial IVIg infusion. The age range was 2 months to 11 years (mean 42.15 ± 38.51 months). More than two-thirds of the cohort was male (n = 27/39). Mucosal involvement was the commonest clinical abnormality for the group, followed by rash. Hemoglobin was significantly lower in the group with coronary artery involvement (P = 0.001). CAA (61.5%), incomplete KD, and atypical features were much more common in infants compared to the rest. Refractoriness to treatment was significantly more common in infants (P = 0.029). A significant proportion of infants with KD had cardiac involvement. Infants were more likely to have IVIg-resistant disease.


Development of a total colonoscopy rat model with endoscopic submucosal injection of the cecal wall.
Experimental models of colorectal tumor require either laparotomy for induction or anastomosis following resection. The long murine cecum avoids the need for an anastomosis, making the cecum the preferred site for induction. This study aimed to evaluate total colonoscopy with submucosal injection of cecal wall (TCWI) in rats in terms of failure rate (FR), complication rate (CR), and reproducibility (R). A bolus of bowel prep was given. Anesthesia was injected intraperitoneally. A video fiberscope (5.9 mm outer diameter, 180/90 degrees up/down bending, 100/100 degrees right/left bending, 103 cm working length, 120 degrees view field, and 2.0 mm channel) allowed for irrigation and suction. Saline 1 ml was injected in the cecal wall through a 4-mm-long, 23-gauge needle placed on a 3-mm wire, resulting in a blister. FR was a failure to reach and inject the cecum. Rats were allowed to recover. CR was measured at necropsy. R was assessed by comparing TCWI time, FR, and CR for three investigators. Sample size of 120 (type I error, 0.05; power, 80%) was based on a pilot study. Data are presented as median (range). A total of two of 122 rats (1.6%) died after prep or anesthesia. Bowel prep resulted in 99.1% evacuation of solid feces. A total of 120 male Sprague-Dawley retired breeders weighing 592 g (range, 349-780) underwent TCWI. Scope depth was 28 cm (range, 20-36). Irrigating fluid was 290 ml (range, 100-600). TCWI time was 7 min (range, 4-28). FR was 4%. In three failed cases, the scope reached the ascending colon. CR was 2%. There were two perforations in the ascending colon. All three operators had similar TCWI time (p = 0.673), FR (p > 0.1), and CR (p > 0.1). A total of 98.3% of rats survived to planned sacrifice. At 48-h necropsy, the injection site was macroscopically identified in 118 rats. A safe and reproducible TCWI rat model has been achieved, which may provide a valuable tool in the future for studies of solid colorectal tumors.


[Past, present and future of zidovudine, the first antiretroviral drug].
The zidovudine (ZDV) was the first drug approved to treat the VIH infection. The ZDV prescriptions have been changing throughout the years. Actually it's known that its efficacy is limited over time, and same substances have been discovered which inhibtes more strongly the VIH replication. However the ZDV appears in majority the combined therapy regimens as a first line drug. Even in same special situations there is no existence of another antiretroviral drugs which have showed utility. In this essay it s revised the mainly studies which have provided new knowledge about the ZDV treatment and it s also considered the possibilities of this drug in the more new therapies against the VIH.


Generation of an attenuated H5N1 avian influenza virus vaccine with all eight genes from avian viruses.
In the face of disease outbreaks in poultry and the potential pandemic threat to humans caused by the highly pathogenic avian influenza viruses (HPAIVs) of H5N1 subtype, improvement in biosecurity and the use of inactivated vaccines are two main options for the control of this disease. Vaccine candidates of influenza A viruses of H5N1 subtype have been generated in several laboratories by plasmid-based reverse genetics with hemagglutinin (HA) and neuraminidase (NA) genes from the epidemic strains of avian viruses in a background of internal genes from the vaccine donor strain of human strains, A/Puerto Rico/8/34 (PR8). These reassortant viruses containing genes from both avian and human viruses might impose biosafety concerns, also may be do if C4/F AIV would be a live attenuated vaccine or cold-adaptive strain vaccine. In order to generate better and safer vaccine candidate viruses, we genetically constructed attenuated reassortant H5N1 influenza A virus, designated as C4/F AIV, by plasmid-based reverse genetics with all eight genes from the avian strains. The C4/F AIV virus contained HA and NA genes from an epidemic strain A/Chicken/Huadong/04 (H5N1) (C4/H5N1) in a background of internal genes derived from a low pathogenic strain of A/Chicken/F/98(H9N2). The reassortant virus was attenuated by removal of the multibasic amino acid motif in the HA gene by mutation and deletion (from PQRERRRKKR (downward arrow) G to PQIETR (downward arrow) G). The intravenous pathogenicity index (IVPI) of C4/F AIV virus was 0, whereas that of the donor virus C4/H5N1 was 3.0. The virus HA titer of C4/H5N1 in the allantoic fluid from infected embryonated eggs was as high as 1:2048. The inactivated vaccine prepared from the reassortant virus C4/F AIV-induced high HI titer in vaccinated chickens and gave 100% protection when challenged with highly pathogenic avian influenza virus of H5N1 subtype.


Conserved ERAD-like quality control of a plant polytopic membrane protein.
The endoplasmic reticulum (ER) of eukaryotic cells serves as a checkpoint tightly monitoring protein integrity and channeling malformed proteins into different rescue and degradation routes. The degradation of several ER lumenal and membrane-localized proteins is mediated by ER-associated protein degradation (ERAD) in yeast (Saccharomyces cerevisiae) and mammalian cells. To date, evidence for the existence of ERAD-like mechanisms in plants is indirect and based on heterologous or artificial substrate proteins. Here, we show that an allelic series of single amino acid substitution mutants of the plant-specific barley (Hordeum vulgare) seven-transmembrane domain mildew resistance o (MLO) protein generates substrates for a postinsertional quality control process in plant, yeast, and human cells, suggesting conservation of the underlying mechanism across kingdoms. Specific stabilization of mutant MLO proteins in yeast strains carrying defined defects in protein quality control demonstrates that MLO degradation is mediated by HRD pathway-dependent ERAD. In plants, individual aberrant MLO proteins exhibit markedly reduced half-lives, are polyubiquitinated, and can be stabilized through inhibition of proteasome activity. This and a dependence on homologs of the AAA ATPase CDC48/p97 to eliminate the aberrant variants strongly suggest that MLO proteins are endogenous substrates of an ERAD-related plant quality control mechanism.


Oxytocin determination by radioimmunoassay. III. Improvement to subpicogram sensitivity and application to blood levels in cyclic cattle.
An improved RIA for measurement of oxytocin in blood is described by using an extraction method with SEP-PAK C18 cartridges, which allows also concentration of the sample, a new antiserum with a higher sensitivity to standard oxytocin and preparation of the standard curve in buffer. The lower limit of assay sensitivity was 0.25 pg/tube, corresponding to 0.25-1.0 pg/ml plasma depending on the amount of plasma extracted. Hence, it was no problem to measure oxytocin basal concentrations in peripheral blood in the range of 0.6-4 pg/ml plasma depending on the stage of the oestrous cycle. The highest oxytocin concentrations occurred during the early and mid-luteal phase. The method has been applied also for samples from women, sheep, pigs and horses. Mean (+/- SD) recovery of oxytocin added to plasma or only buffer after extraction was 71.3 +/- 8.1%, and the coefficient of variation (CV) = 11.4% (n = 27 assays). The intra-assay CV of two control samples was 7.9 +/- 2.8 and 7.8 +/- 2.4% (n = 17 assays). The inter-assay CV of 5 control samples with low and high oxytocin concentrations varied between 10.8 +/- 17.3% (n = 25 assays). The 50% intercept was 2.5 +/- 0.3 pg, CV = 11.3% (n = 29 assays).


Initial control of chemotherapy-induced nausea and vomiting in patient quality of life.
The side effects commonly experienced by patients receiving chemotherapy for the treatment of cancer can challenge many aspects of daily life. Nausea and vomiting, the most common side effects reported by patients, affect the ability to continue with usual life activities and, thus have a pronounced impact on quality of life. This paper reviews studies of the impact of nausea and emesis on quality of life, and highlights the importance of prevention of these side effects by presenting new data on how persistent uncontrolled nausea and vomiting can be. The Morrow Assessment of Nausea and Emesis (MANE) was used to collect information on symptoms experienced by consecutive patients starting chemotherapy between September 1987 and December 1995 at any of 18 geographically diverse member sites of the University of Rochester Cancer Center Community Clinical Oncology Program. Data from 1,413 patients were collected after each of four successive chemotherapy treatments. Reported incidences of posttreatment nausea and posttreatment vomiting after the first treatment were 59.4% and 28.6%, respectively. Occurrence of nausea/vomiting at the first treatment was a strong predictor of nausea/vomiting at later treatments. Of the 839 patients reporting initial nausea, 763 (90.9%) reported nausea at at least one subsequent treatment, and approximately 59% reported nausea after all three subsequent treatments. Fewer than half (45.6%) of the patients who had no nausea at the first treatment developed it later. The majority (72.0%) of patients reporting vomiting at the first treatment also reported subsequent vomiting, 30.7% of whom experienced emesis at all remaining treatments. Conversely, 76.2% of patients who were emesis-free at the first treatment remained so for all later treatments. These findings show a continuing need for further progress in controlling nausea and vomiting, and demonstrate the importance of aggressive nausea/vomiting control at the first treatment. In addition, more emphasis on controlling chemotherapy-induced nausea after its initial occurrence is necessary.


Evaluation of two new enzyme immunoassay reagents for diagnosis of histoplasmosis in a cohort of clinically characterized patients.
The performance characteristics of the recently available analyte-specific reagent based enzyme immunoassay (ASR-EIA) and in vitro diagnostic (IVD) kit for urine Histoplasma antigen detection were evaluated in a cohort of 50 clinically characterized patients with histoplasmosis and 50 control patients. Overall sensitivity and specificity of the ASR-EIA were significantly improved compared with those of the IVD kit (sensitivity 72% vs. 22%, P<.001, specificity 98% vs. 84%, P = .014). Fourteen specimens from patients with clinically characterized histoplasmosis (five with pulmonary histoplasmosis and nine with progressive disseminated histoplasmosis) were falsely negative by ASR-EIA. All 10 specimens from patients with severe symptoms of progressive disseminated histoplasmosis were positive by ASR-EIA, although the average reading value of these 10 specimens was not significantly different from that of others with positive results. Compared to the MiraVista antigen assay, both the IVD kit and the ASR-EIA were significantly less sensitive in detecting Histoplasma antigen in the urine of patients with histoplasmosis. The ASR-EIA and MiraVista assay had comparable specificity. In conclusion, the ASR-EIA has improved performance compared with the IVD kit in the detection of Histoplasma antigen in the urine. However, users should be aware of the potential for false negative results using the currently recommended cutoff value.


Implementation of cancer prevention guidelines in clinical practice.
Data from several sources, including consumer surveys, physician surveys, and medical record audits, indicate that consumers do not receive cancer screening tests as recommended by the National Cancer Institute, the American Cancer Society, and the U.S. Preventive Services Task Force. Performance rates are consistently below published standards for all tests except Pap tests. Major reasons physicians do not perform the recommended tests include physician forgetfulness, disagreement with recommendations, lack of time, and patient refusal. Physicians also tend to overestimate their own performance rates. Barriers to screening test performance can be categorized into patient factors, physician factors, test factors, and health care delivery system factors. Interventions, such as computerized reminder systems, physician audits with feedback, and patient education and reminders, can be effective in promoting performance of such screening. Interventions that target both physician and patient may be particularly effective.


Myocardial deformation analysis in contrast echocardiography: first results using two-dimensional cardiac performance analysis.
Contrast echocardiography (CE) provides closer agreement with magnetic resonance imaging (MRI) for left ventricular (LV) volumes and ejection fraction (EF) than noncontrast echocardiography. However, the feasibility and role of myocardial deformation analysis on contrast echocardiographic images have not been well established. The aim of this study was to assess the feasibility of deformation analysis on CE using a new software tool that provides simultaneous measurements for LV volumes and EF. Data from 52 patients who were recruited for the Alberta Heart Failure Etiology and Analysis Research Team Study (34 men; mean age, 64 ± 9 years) and underwent CE and MRI were considered. Contrast bolus injections were administered for optimal endocardial definition. Offline LV volume analysis was performed by standard manual tracing. A single frame was traced manually for two-dimensional (2D) cardiac performance analysis (CPA), which automatically calculated LV volumes, EF, and global longitudinal strain (GLS). Volumes obtained with 2D CPA were compared with those measured with standard CE and MRI. GLS from noncontrast echocardiographic recordings was also calculated with 2D CPA and compared with CE-derived and MRI-derived GLS. Tracing of contrast echocardiographic images with 2D CPA was possible in 49 out of 52 patients, and measurements correlated well with standard CE and MRI (EF: r = 0.93, P < .001, and r = 0.85, P < .001, respectively). Mean GLS from noncontrast echocardiographic and contrast echocardiographic recordings was -13.4 ± 5.8 and -15.3 ± 4.64, respectively (P = .056), and the latter correlated well with MRI-derived GLS (r = 0.78 vs 0.81, respectively). Simultaneous volumetric and deformation analysis on contrast echocardiographic recordings is feasible and reproducible. While volumes and EF obtained with the new software compare well with those obtained from standard CE and MRI, GLS from CE shows a good correlation with strain measured with MRI.


[Critical analysis of respiratory infectious disease investigations related to children attending day care centers]
OBJECTIVE: Social-economic and cultural changes are increasingly leading families to share the children's care and education with daycare centers. Nevertheless, this practice is being widely criticized. Medical literature, for example, points to the high risk of illnesses in these environments. Searching to assess theoretical- methodological aspects underlying disease/daycare research, a bibliography review was done. METHOD: The last five years production was reviewed, through Current Contents, Med-Line and Lilacs, using creche / daycare center as keywords. RESULTS: 665 papers were obtained and organized in subgroups. Selecting the group of papers on respiratory infectious diseases, 34 papers were obtained, being 22 of them empirical, and 12 theoretical or review article, etc. The empirical studies used large populations; data were collected by interviews, medical reports and laboratory analyses; 50% were cross-sectional. Disease/daycare relation was obtained mainly by quantitative analysis, based on a multivariate model and logistic regression. Seventy per cent concluded that attending daycare implies an illness high risk, especially for youngers than 2 years. The papers with a preventive character, and-in more subtler ways - 50% of the others point alternative solutions for this clinical and social problem (vaccination, immunity stimulation, environmental organization). CONCLUSION: The structure and the discussion of the majority of the studies seem to be guided by implicit assumptions, based on conceptions of an ideal situation of care (at house, by the mother). The results are legitimized by large populations and sophisticated statistic systems. Nevertheless, they express a prediction risk, which highlights static aspects, not the dynamic ones, of health/ disease processes. At last, they treat a social problem on clinical individual assumptions.


Synchronization of intermittent behavior in ensembles of multistable dynamical systems.
We propose a methodology to analyze synchronization in an ensemble of diffusively coupled multistable systems. First, we study how two bidirectionally coupled multistable oscillators synchronize and demonstrate the high complexity of the basins of attraction of coexisting synchronous states. Then, we propose the use of the master stability function (MSF) for multistable systems to describe synchronizability, even during intermittent behavior, of a network of multistable oscillators, regardless of both the number of coupled oscillators and the interaction structure. In particular, we show that a network of multistable elements is synchronizable for a given range of topology spectra and coupling strengths, irrespective of specific attractor dynamics to which different oscillators are locked, and even in the presence of intermittency. Finally, we experimentally demonstrate the feasibility and robustness of the MSF approach with a network of multistable electronic circuits.


Relationships of quality-of-life measures to long-term lifestyle and drug treatment in the Treatment of Mild Hypertension Study.
To compare 5 antihypertensive drugs and placebo for changes in quality of life (QL). To assess the relationship of lifestyle factors and change in lifestyle factors to QL in participants with stage I diastolic hypertension. The Treatment of Mild Hypertension Study (TOMHS) was a randomized, double-blind, placebo-controlled clinical trial with minimum participant follow-up of 4 years. It was conducted at 4 hypertension screening and treatment academic centers in the United States. The cohort consisted of 902 men and women with hypertension, aged 45 to 69 years, with diastolic blood pressures less than 100 mm Hg. Informed consent was obtained from each participant after the nature of the procedures had been fully explained. Sustained nutritional-hygienic intervention was administered to all participants to reduce weight, to reduce dietary sodium and alcohol intake, and to increase physical activity. Participants were randomized to take (1) acebutolol (n = 132); (2) amlodipine maleate (n = 131); (3) chlorthalidone (n = 126); (4) doxazosin mesylate (n = 134); (5) enalapril maleate (n = 135); or placebo (n = 234). Changes in 7 QL indexes were assessed based on a 35-item questionnaire: (1) general health; (2) energy or fatigue; (3) mental health; (4) general functioning; (5) satisfaction with physical abilities; (6) social functioning; and (7) social contacts. At baseline, higher QL was associated with older age, more physical activity, lower obesity level, male gender, non-African American race, and higher educational level. Improvements in QL were observed in all randomized groups, including the placebo group during follow-up; greater improvements were observed in the acebutolol and chlorthalidone groups and were evident throughout follow-up. The amount of weight loss, increase in physical activity, and level of attained blood pressure control during follow-up were related to greater improvements in QL. In patients with stage I hypertension, antihypertensive treatment with any of 5 agents used in TOMHS does not impair QL. The diuretic chlorthali-done and the cardioselective beta-blocker acebutolol appear to improve QL the most. Success with lifestyle changes affecting weight loss and increase in physical activity relate to greater improvements in QL and show that these interventions, in addition to contributing to blood pressure control, have positive effects on the general well-being of the individual.


The synthesis of export-defective proteins can interfere with normal protein export in Escherichia coli.
We have analyzed the kinetics of maturation for certain bacterial envelope proteins in Escherichia coli strains that are also concomitantly producing an export-defective protein. Our data indicate that proteins with defective signal peptides, rendered nonfunctional by either point mutation or deletion, interfere with the normal export of other envelope proteins. Expression of interference requires that the interfering protein: (i) exhibit a major export defect; (ii) be synthesized at a high rate; and (iii) be actively synthesized at the time interference is being measured. The latter data suggest that interference is a cotranslational process. Intragenic or extragenic suppression of the export defect exhibited by the interfering protein relieves interference in a manner that is directly related to strength of suppression. These and additional data suggest that interference occurs at a very early step in the secretory process. We interpret these results to indicate that proteins with defective signal peptides are still recognized as proteins destined for secretion and are, therefore, at least transiently incorporated into the cell's secretory pathway. The incorporation of an export-defective protein into the secretory pathway disrupts the normal protein traffic from the cytoplasm to the various extracellular compartments.


Aedes albopictus in Muzquiz city, Coahuila, Mexico.
Aedes albopictus was found in M. Muzquiz, Coahuila, Mexico, in the summer of 1993. Aedes albopictus larvae were collected in tires and bottles at low density. Aedes aegypti, Toxorhynchites theobaldi, Culex quinquefasciatus, and Culiseta incidens were collected in the same containers.


[Problems in medical care for patients with cardiovascular diseases].
Despite showing that cardiovascular disease mortality in Russia is declining evaluation of medical care for cardiovascular patients, using different information sources, revealed a lot of problems in this field need it's solving. Together with modernization of medical services it is urgently necessary to transform the medical education, including post graduate, information support for medical professionals in field of modern medical technologies, using in countries with low mortality rates, creation the conditions for regular updating professional knowledge. It is necessary to reconsider formal criteria for medical care evaluation, especially taking into account co morbidity of cardiovascular diseases. Our data illustrate that social disparities influence on outcomes of diseases and so for further decreasing mortality rates and increasing the life expectancy adequate treatment must be provide for all patients independently of their material wellbeing.


A Case of Lingua Villosa Nigra (Black Hairy Tongue) in a 3-Month-Old Infant.
BACKGROUND Black hairy tongue is a self-limiting, usually asymptomatic, benign lesion that most often affects men and people aged over 30-40 years. The lesion is extremely rare among newborns and infants. Lingua villosa nigra is characterized by the presence of elongated filiform papillae of the dorsum of the tongue, which gives a hairy appearance. The overgrown papillae can accumulate fungi, bacteria, pigments originating from food, or any other debris that can contribute to the discoloration of the tongue. The prognosis for black hairy tongue is positive. Gentle tongue brushing or scraping as well as the elimination of predisposing factors usually leads to cessation of the lesion. CASE REPORT A generally healthy, exclusively breastfed 3-month-old female baby was admitted to the Oral Pathology Department of Wrocław Medical University due to a persistent tongue lesion. Intraorally, dark, blackish, elongated tongue papillae were observed. Three weeks earlier, the baby's pediatrician had diagnosed thrush and prescribed systemic antifungal treatment with Nystatinum, without prior mycological examination. The lesion did not resolve and the girl was referred to the Department of Oral Pathology. A meticulous medical and dietary interview revealed that since the 28th day of life the baby had been supplemented with vitamin C. This, together with an intraoral examination, led to the diagnosis of black hairy tongue. The lesion disappeared partially after 4 weeks of tongue brushing. CONCLUSIONS To prevent infants from undergoing persistent and unnecessary treatment (topical or systemic drugs) or additional diagnostic procedures, such as biopsy, it is essential to be familiar with the characteristics of lingua villosa nigra as well as its origin and management. The consideration of this condition is invaluable for babies' health and safety.


Antibodies against GABA and glutamate label neurons with morphologically distinct synaptic vesicles in the locust central nervous system.
Antibodies raised against GABA and glutamate were used to stain sections through locust thoracic ganglia for light and electron microscopy. Using a peroxidase-antiperoxidase method for light microscopy, the GABA antibody was shown to label inhibitory motor neurons thought to use GABA as their neurotransmitter, and the glutamate antibody to label excitatory motor neurons thought to use glutamate. An immunogold method was used to reveal labelled neuropilar processes in the electron microscope. Each antibody specifically labels a particular population of processes. With the GABA antibody, labelling is equally clear whether the processes concerned contain synaptic vesicles or not and is strongly contrasted against very low background levels. With the glutamate antibody, most processes show some affinity for the antibody, probably reflecting the presence of metabolic glutamate, however one population can be clearly distinguished by the presence of a much greater density of gold particles over synaptic vesicles. In the locust it appears, therefore, that the antibody can distinguish clearly between the metabolic and neurotransmitter pools of glutamate. It has been proposed that synaptic vesicles in GABAergic neurons have a different shape to those in glutamatergic neurons. This was supported by the electron microscope immunocytochemistry. Those showing GABA-like immunoreactivity contain predominantly pleomorphic agranular vesicles approximately 21 x 30 nm in diameter. Those showing glutamate-like immunoreactivity contain round agranular vesicles of about 38 nm in diameter. The GABA antibody appears to label all processes containing pleomorphic agranular vesicles. By contrast, some processes containing round agranular vesicles are not labelled by the glutamate antibody, even though the vesicles they contain are statistically identical in size to those in labelled profiles. With neither antibody was the labelling of glial cells greater than the background level.


Comparison of Percentage Prolonged Times to Tracheal Extubation Between a Japanese Teaching Hospital and One in the United States, Without and With a Phase I Postanesthesia Care Unit.
Prolonged times to tracheal extubation are those from end of surgery (dressing on the patient) to extubation 15 minutes or longer. They are so long that others in the operating room (OR) generally have exhausted whatever activities can be done. They cause delays in the starts of surgeons' to-follow cases and are associated with longer duration workdays. Anesthesiologists rate them as being inferior quality. We compare prolonged times to extubation between a teaching hospital in the United States with a phase I postanesthesia care unit (PACU) and a teaching hospital in Japan without a PACU. Our report is especially important during the coronavirus disease 2019 (COVID-19) pandemic. Anesthesiologists with some patients undergoing general anesthetics and having initial PACU recovery in the ORs where they had surgery can learn from the Japanese anesthesiologists with all patients recovering in ORs. The historical cohort study included all patients undergoing gynecological surgery at a US hospital (N = 785) or Japanese hospital (N = 699), with the time from OR entrance to end of surgery of at least 4 hours. The mean times from end of surgery to OR exit were slightly longer at the US hospital than at the Japanese hospital (mean difference 1.9 minutes, P < .0001). The mean from end of surgery to discharge to surgical ward at the US hospital also was longer (P < .0001), mean difference 2.2 hours. The sample standard deviations of times from end of surgery until tracheal extubation was 40 minutes for the US hospital versus 4 minutes at the Japanese hospital (P < .0001). Prolonged times to tracheal extubation were 39% of cases at the US hospital versus 6% at the Japanese hospital; relative risk 6.40, 99% confidence interval (CI), 4.28-9.56. Neither patient demographics, case characteristics, surgeon, anesthesiologist, nor anesthesia provider significantly revised the risk ratio. There were 39% of times to extubation that were prolonged among the patients receiving neither remifentanil nor desflurane (all such patients at the US hospital) versus 6% among the patients receiving both remifentanil and desflurane (all at the Japanese hospital). The relative risk 7.12 (99% CI, 4.59-11.05) was similar to that for the hospital groups. Differences in anesthetic practice can facilitate major differences in patient recovery soon after anesthesia, useful when the patient will recover initially in the OR or if the phase I PACU is expected to be unable to admit the patient.


Structure, Function and Regulation of Outer Membrane Proteins Involved in Drug Transport in Enterobactericeae: the OmpF/C - TolC Case.
Antibiotic translocation across membranes of Gram-negative bacteria is a key step for the activity on their specific intracellular targets. Resistant bacteria control their membrane permeability as a first line of defense to protect themselves against external toxic compounds such as antibiotics and biocides. On one hand, resistance to small hydrophilic antibiotics such as ß-lactams and fluoroquinolones frequently results from the « closing » of their way in: the general outer membrane porins. On the other hand, an effective way out for a wide range of antibiotics is provided by TolC-like proteins, which are outer membrane components of multidrug efflux pumps. Accordingly, altered membrane permeability, including porin modifications and/or efflux pumps' overexpression, is always associated to multidrug resistance (MDR) in a number of clinical isolates. Several recent studies have highlighted our current understanding of porins/TolC structures and functions in Enterobacteriaceae. Here, we review the transport of antibiotics through the OmpF/C general porins and the TolC-like channels with regards to recent data on their structure, function, assembly, regulation and contribution to bacterial resistance. Because MDR strains have evolved global strategies to identify and fight our antibiotic arsenal, it is important to constantly update our global knowledge on antibiotic transport.


Pregnancy as foreground in cystic fibrosis carrier testing decisions in primary care.
Cystic fibrosis carrier testing (CFCT) is among the first of the DNA tests offered prenatally in primary care settings. This paper from a descriptive qualitative study describes the influence of pregnancy in CFCT decisions by women receiving community-based prenatal care. Twenty-seven women receiving prenatal care in Midwestern U.S. primary care clinics completed semistructured interviews. Audiotaped interviews were analyzed using content analysis. Participants described decision-making influences and strategies from the perspective of "being pregnant." Patterns of attitudes and beliefs include (1) dealing with emotions, (2) pregnancy is natural, and (3) thinking about the baby. Strategies in the decision-making process included (1) reducing stress, (2) choosing what is relevant, (3) doing everything right, (4) wanting to be prepared, (5) delaying information, and (6) trusting God. While other factors were mentioned by some women, major themes reflect the influence of currently being pregnant on the decision-making process. These findings suggest that pregnancy is a powerful influence on the decision-making process and may not be the optimal time to make fully informed decisions regarding genetic carrier testing. Further understanding of factors influencing the genetic testing decision-making process is needed. Offering CFCT prior to conception is advocated.


Cardiolipin, conformation, and respiratory complex-dependent oligomerization of the major mitochondrial ADP/ATP carrier in yeast.
The phospholipid cardiolipin has pleiotropic structural and functional roles that are collectively essential for mitochondrial biology. Yet, the molecular details of how this lipid supports the structure and function of proteins and protein complexes are poorly understood. To address this property of cardiolipin, we use the mitochondrial adenosine 5'-diphosphate/adenosine 5'-triphosphate carrier (Aac) as a model. Here, we have determined that cardiolipin is critical for both the tertiary and quaternary assembly of the major yeast Aac isoform Aac2 as well as its conformation. Notably, these cardiolipin-provided structural roles are separable. In addition, we show that multiple copies of Aac2 engage in shared complexes that are largely dependent on the presence of assembled respiratory complexes III and IV or respiratory supercomplexes. Intriguingly, the assembly state of Aac2 is sensitive to its transport-related conformation. Together, these results expand our understanding of the numerous structural roles provided by cardiolipin for mitochondrial membrane proteins.


A MEG investigation of somatosensory processing in the rhesus monkey.
The use of minimally and non-invasive neuroimaging methods in animal models has sharply increased over the past decade. Such studies have enhanced understanding of the neural basis of the physical signals quantified by these tools, and have addressed an assortment of fundamental and otherwise intractable questions in neurobiology. To date, these studies have almost exclusively utilized positron-emission tomography or variants of magnetic resonance based imaging. These methods provide largely indirect measures of brain activity and are strongly reliant on intact vasculature and normal blood-flow, which is known to be compromised in many clinical conditions. The current study provides the first demonstration of whole-head magnetoencephalography (MEG), a non-invasive and direct measure of neuronal activity, in a rhesus monkey, and in the process supplies the initial data on systems-level dynamics in somatosensory cortices. An adult rhesus monkey underwent three separate studies of tactile stimulation on the pad of the right second or fifth digit as whole-head MEG data were acquired. The neural generators of the primary neuromagnetic components were localized using an equivalent-current-dipole model. Second digit stimulation produced an initial cortical response peaking approximately 16 ms after stimulus onset in the contralateral somatosensory cortices, with a later response at approximately 96 ms in an overlapping or nearby neural area with a roughly orthogonal orientation. Stimulation of the fifth digit produced similar results, the main exception being a substantially weaker later response. We believe the 16 ms response is likely the monkey homologue of the human M50 response, as both are the earliest cortical response and localize to the contralateral primary somatosensory area. Thus, these data suggest that mechanoreception in nonhuman primates operates substantially faster than that in adult humans. More broadly, these results demonstrate that it is feasible to use current human whole-head MEG instrumentation to record neuromagnetic responses in adult rhesus monkeys. Nonhuman primate models of human disease provide the closest phylogenetic link to humans. The present, non-invasive imaging study could promote exciting translational integration of invasive animal studies and non-invasive human studies, allowing experimentally induced deficits and pharmacological treatments to be interpreted in light of resulting brain network interactions.


Calcium [corrected] channel blockers reduce angiotensin II-induced superoxide generation and inhibit lectin-like oxidized low-density lipoprotein receptor-1 expression in endothelial cells.
Calcium channel blockers have been shown to limit the progression of atherosclerosis and decrease the incidence of cardiovascular events. To investigate vasoprotective effects beyond the blood pressure-lowering effects of these agents, amlodipine (10(-6) mol/) and manidipine (10(-6) mol/l) were used to pretreat angiotensin (Ang) II-stimulated rat cultured aortic endothelial cells. A 3-h period of Ang II treatment enhanced superoxide generation and the expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase protein, as detected by dihydroethidium staining and Western blotting, respectively. Pretreatment with amlodipine or manidipine attenuated the increased production of superoxide and the overexpression of NADPH oxidase. The enhanced expression of heme oxygenase-1 (HO-1) mRNA induced by Ang II was further increased by amlodipine, whereas pretreatment with manidipine led to a reduction in the expression of HO-1. Furthermore, Ang II increased vascular cell adhesion molecule-1 (VCAM-1), intracellular adhesion molecule-1 (ICAM-1), and monocyte chemoattractant protein-1 (MCP-1) mRNA levels, as determined by reverse transcription (RT)-polymerase chain reaction (PCR). Pretreatment with either amlodipine or manidipine decreased the overexpression of VCAM-1, ICAM-1, and MCP-1. We also demonstrated that amlodipine or manidipine prevented the Ang II-induced increase in lectin-like oxidized low-density lipoprotein receptor1 (LOX-1) content, thereby restoring control levels. These observations showed that amlodipine and manidipine reduced superoxide generation by the inhibition of the overexpression of NADPH oxidase in Ang II-stimulated endothelial cells. Such antioxidant effects of these agents might in turn have led to a decrease in the expression of VCAM-1, ICAM-1 and MCP-1. The salutary effects of calcium channel blockers in atherogenesis include the inhibition of the expression of LOX-1.


Adaptative capacity of mitochondrial biogenesis and of mitochondrial dynamics in response to pathogenic respiratory chain dysfunction.
Cellular energy homeostasy relies on mitochondrial plasticity, the molecular determinants of which are multiple. Yet, the relative contribution of and possible cooperation between mitochondrial biogenesis and morphogenesis to cellular energy homeostasy remains elusive. Here we analyzed the adaptative capacity of mitochondrial content and dynamics in muscle biopsies of patients with a complex IV defect, and in skin fibroblasts challenged with complex IV inhibition. We observed a biphasic variation of the mitochondrial content upon complex IV inhibition in muscle biopsies and in skin fibroblasts. Adjustment of mitochondrial content for respiratory maintenance was blocked by using a dominant negative form of CREB (CREB-M1) and by L-NAME, a blocker of NO production. Accordingly, cells treated with KCN 6 μM showed higher levels of phospho-CREB, PGC1α mRNA, eNOS mRNA, and mtTFA mRNA. We also observed the increased expression of the fission protein DRP1 during fibroblasts adaptation, as well as mitochondrial ultrastructural defects indicative of increased fission in patients muscle micrographs. Accordingly, the expression of a dominant negative form of DRP1 (K38A mutant) reduced the biogenic response in fibroblasts challenged with 6 μM KCN. Our findings indicate that mitochondrial biogenesis and mitochondrial fission cooperate to promote cellular adaptation to respiratory chain inhibition. Our data show for the first time that DRP1 intervenes during the initiation of the mitochondrial adaptative response to respiratory chain defects. The evidenced pathway of mitochondrial adaptation to respiratory chain deficiency provides a safety mechanism against mitochondrial dysfunction.


Receptor occupancy regulates Ca2+ entry and intracellular Ca2+ redistribution in activated human platelets.
Fura-2-loaded human platelets were used to study Ca2+ release from intracellular compartments, as well as Ca2+ influx from the extracellular space. We investigated the response towards the endoperoxide/thromboxane-receptor agonist. U46619, and the inhibitor of the endoplasmic-reticulum Ca(2+)-ATPase, thapsigargin. U46619 dose-dependently depleted intracellular Ca2+ stores, followed by active sequestration of released Ca2+. Ca2+ influx induced by U46619 largely relies on receptor occupancy. Removing the thromboxane analogue from its receptor by using the endoperoxide/thromboxane-receptor antagonist BM 13177 largely blunted U46619-mediated Ca2+ influx. The Ca(2+)-ATPase inhibitor thapsigargin evoked a gradual rise in intracellular Ca2+, which was potentiated by a preceding activation of platelets with the receptor agonist U46619. This agonist-sensitizing effect also depends on receptor occupancy. Removing U46619 from its receptor by addition of the endoperoxide/thromboxane-receptor antagonist BM13177 suppressed the sensitizing effect completely. Furthermore, interrupting downstream receptor signalling events by raising intracellular levels of cyclic nucleotides (cyclic AMP, cyclic GMP) again suppressed the U46619-sensitizing effect on thapsigargin-induced Ca2+ release. This study indicates that the process of Ca2+ release followed by resequestration in response to a platelet agonist by its own is not sufficient to produce the sensitizing effect. Rather, a continuously occupied receptor triggering sustained downstream signalling events seems to be required for sensitization. The presence of a receptor agonist may induce an increased cycling of Ca2+ between the agonist-responsive and the thapsigargin-dischargeable compartment, leading to faster and more intense accumulation of Ca2+ in the cytosolic compartment after inhibition of the Ca(2+) ATPase. Suggestively, receptor occupancy increases the Ca(2+)-releasing potency of thapsigargin by coupling the thapsigargin-sensitive Ca(2+)-storing compartments with an agonist-responsive compartment that exhibits a high leakage rate in stimulated platelets.


The regulation of exopolysaccharide production is important at two levels of nodule development in Rhizobium meliloti.
We show that two exopolysaccharide overproducing Tn5 mutants of Rhizobium meliloti, exoR and exoS, have distinct symbiotic defects. While the exoR mutant is unable to colonize nodules, the exoS mutant retains that ability but varies in its ability to produce nitrogen-fixing nodules. We correlate these defects with different degrees of exopolysaccharide overproduction and growth impairment. We further show that the exoR mutant is able to enter developing infection threads but is unable to invade nodule cells. The exoR mutant gives rise to spontaneous pseudorevertants containing second-site suppressor mutations that decrease exopolysaccharide synthesis. These pseudorevertants form nitrogen-fixing nodules. Although the suppressor mutations have the opposite effect on exopolysaccharide production compared to the exoS::Tn5 mutation, they consistently map to the exoS::Tn5 region and belong to the same genetic complementation group as defined by transposon insertion mutations. The effect of the suppressor mutations on exopolysaccharide production is correlated with effects on the expression of exo genes involved in exopolysaccharide synthesis. Finally, we provide evidence that the exoR gene is not required for the regulation of exopolysaccharide synthesis by ammonia.


Pharmaceutical interventions in the emergency department: cost-effectiveness and cost-benefit analysis.
It has been shown that pharmacists, as members of multidisciplinary patient care teams, can decrease the number of medicine errors. The objective of the present study was to analyse pharmaceutical interventions (PI) in emergency departments, to assess their clinical relevance, the cost-effectiveness and the potential economic benefits. We designed a 5-month observational prospective study of PI in the emergency department (ED) of a 330-bed hospital in Spain. We analysed PI made by a pharmacist during a period of 3 hours a day from Monday to Friday in the ED, and classified detected medication errors according to their relevance and severity using the National Coordinating Council for Medication Error Reporting and Prevention (NCC MERP) severity index, and whether or not the drug involved was on the High-Alert Medications Institute for Safe Medication Practices (ISMP) list. We used statistical analysis to study the relationship between the relevance of PI and age, gender, the number of interventions per patient, and whether or not the drug was on the High-Alert Medications ISMP list. We also estimated the incremental cost incurred for each PI (cost-effectiveness) and the potential economic benefits (cost-benefit). A total of 529 interventions for 390 patients (median age 72.7±8.6 years, 53.1% women) were performed during the study period, representing 1.4 interventions per patient with an acceptance rate of 84.9%. Of all potential medication errors, 112 (21.2%) were related to drugs on the High-Alert Medications ISMP list, and using the NCC MERP severity index, we found that 150 (28.3%) of those errors could cause harm. We also found a relationship between patients on high-risk medications according to the ISMP and the relevance of PI. Finally, this study showed an incremental cost for each PI of 20.23 Euros and a cost-benefit ratio of 3.46 Euros per intervention. These results show that clinical pharmacist can positively identify and reduce medication errors and costs associated, considering the number of interventions observed and those of clinical relevance. Based on these results, drug safety therapy in the ED can be improved by the revision of prescriptions by a clinical pharmacist.


Insulin resistance is associated with tissue-specific regulation of HIF-1α and HIF-2α during mild chronic intermittent hypoxia.
Chronic intermittent hypoxia (CIH) is a feature of obstructive sleep apnea (OSA). Whereas clinical studies have demonstrated the association between OSA and insulin resistance, the molecular mechanisms behind it are still unknown. Herein we investigated the effect of mild CIH on insulin sensitivity and we evaluated the changes in insulin and HIF signaling pathways that occur in CIH-induced insulin resistance. We showed that mild CIH obtained by 5/6 hypoxic (5%O2) cycles/h, 10.5h/day during 28 and 35 days increased arterial blood pressure. Insulin resistance and insulinemia increased with CIH duration, being significantly different after 35 days of CIH. Thirty-five days of CIH decreased insulin receptor expression and phosphorylation in skeletal muscle and adipose tissue, but not in the liver. Conversely, Glut2 expression increased in the liver of CIH-animals. Thirty-five days of CIH up-regulated HIF-1α in the liver and down-regulated HIF-1α and HIF-2α in skeletal muscle. We concluded that the effect of CIH on insulin sensitivity and signaling is time-dependent and is associated with changes in HIF signaling in insulin-sensitive tissues.


The structural and gene expression hypotheses in laminopathic diseases-not so different after all.
Laminopathies are a diverse group of rare diseases with various pathologies in different tissues, which are linked to mutations in the LMNA gene. Historically, the structural disease model proposed mechanical defects of the lamina and nuclear fragility, the gene expression model impairment of spatial chromatin organization and signaling pathways as underlying mechanisms leading to the pathologies. Exciting findings in the past few years showing that mechanical forces are directly transmitted into the nucleus, where they affect chromatin organization and mechanoresponsive signaling molecules, have led to a revised concept of an integrative unified disease model, in which lamin-mediated pathways in mechanotransduction and chromatin regulation are highly interconnected and mutually dependent. In this Perspective we highlight breakthrough findings providing new insight into lamin-linked mechanisms of mechanotransduction and chromatin regulation and discuss how a combined and interrelated impairment of these functions by LMNA mutations may impair the complex mechanosignaling network and cause tissue-specific pathologies in laminopathies.


VP2-segment sequence analysis of some isolates of bluetongue virus recovered in the Mediterranean basin during the 1998-2003 outbreak.
The complete nucleotide sequences of the VP2 segments of bluetongue virus (BTV) isolates recovered from Italy, Greece and Israel, from 1998 to 2003, were determined. Phylogenetic analysis of these sequences, those from related viruses and the South African vaccine strains, were used to determine the probable geographic origin of BTV incursions into Italy. Results indicated that viruses from each of the four serotypes isolated in Italy (2, 4, 9 and 16) possibly had a different origin. Analysis of the bluetongue virus serotype 2 (BTV-2) isolates gave evidence that this serotype probably moved from Tunisia. BTV-4 results showed probable incursion from the southwest and not from Greece or Israel. BTV-9 isolates clearly have an eastern origin (most probably Greece), whereas BTV-16 isolates are indistinguishable from the BTV-16 live attenuated vaccine strain. The phylogenetic findings were supported by polyacrylamide gel electrophoresis (PAGE) analysis of the complete amplified genome of each isolate except for BTV-16 Italian field isolate, which showed a slightly different PAGE profile. A combination of the complete VP2 sequencing and PAGE analysis of complete genomes, allowed not only phylogenetic analysis, but also vaccine detection and assessment of reassortment events.


Stratification of complexity: the Risk Adjustment for Congenital Heart Surgery-1 method and the Aristotle Complexity Score--past, present, and future.
Meaningful evaluation of quality of care must account for variations in the population of patients receiving treatment, or "case-mix". In adult cardiac surgery, empirical clinical data, initially from tens of thousands, and more recently hundreds of thousands of operations, have been used to develop risk-models, to increase the accuracy with which the outcome of a given procedure on a given patient can be predicted, and to compare outcomes on non-identical patient groups between centres, surgeons and eras. In the adult cardiac database of The Society of Thoracic Surgeons, algorithms for risk-adjustment are based on over 1.5 million patients undergoing isolated coronary artery bypass grafting and over 100,000 patients undergoing isolated replacement of the aortic valve or mitral valve. In the pediatric and congenital cardiac database of The Society of Thoracic Surgeons, 61,014 operations are spread out over greater than 100 types of primary procedures. The problem of evaluating quality of care in the management of pediatric patients with cardiac diseases is very different, and in some ways a great deal more challenging, because of the smaller number of patients and the higher number of types of operations. In the field of pediatric cardiac surgery, the importance of the quantitation of the complexity of operations centers on the fact that outcomes analysis using raw measurements of mortality, without adjustment for complexity, is inadequate. Case-mix can vary greatly from program to program. Without stratification of complexity, the analysis of outcomes for congenital cardiac surgery will be flawed. Two major multi-institutional efforts have attempted to measure the complexity of pediatric cardiac operations: the Risk Adjustment in Congenital Heart Surgery-1 method and the Aristotle Complexity Score. Both systems were derived in large part from subjective probability, or expert opinion. Both systems are currently in wide use throughout the world and have been shown to correlate reasonably well with outcome. Efforts are underway to develop the next generation of these systems. The next generation will be based more on objective data, but will continue to utilize subjective probability where objective data is lacking. A goal, going forward, is to re-evaluate and further refine these tools so that, they can be, to a greater extent, derived from empirical data. During this process, ideally, the mortality elements of both the Aristotle Complexity Score and the Risk Adjustment in Congenital Heart Surgery-1 methodology will eventually unify and become one and the same. This review article examines these two systems of stratification of complexity and reviews the rationale for the development of each system, the current use of each system, the plans for future enhancement of each system, and the potential for unification of these two tools.


[Significance of 2-hour blood glucose after standardized steamed bread meal in diabetic screening].
To explore the significance of 2-hour blood glucose after standardized steamed bread meal (SB-2 hBG) in diabetic screening. A retrospective study was conducted for diabetic screening data of annual check-up at PLA General Hospital from May 1996 to June 2002. And 100 g standardized steamed bread meal test was performed for non-diabetic subjects. Those subjects with SB-2 h BG ≥ 7.2 mmol/L underwent a 75 g oral glucose tolerance test (OGTT) within 2 weeks to determine whether the diagnosis of diabetes mellitus (DM) could be established (WHO, 1985, 1999, Diagnostic Criteria for Diabetes). By extracting the data for 7 consecutive years, we analyzed the significance and the cut-off point of SB-2 hBG in the diagnosis of DM and investigated the changes of blood glucose curves in different glucose tolerance status after different glucose loading tests. A total of 3 343 subjects with complete information were recruited. There were 3 101 males and 242 females with an age range of 40-94 years. According to the results of OGTT, 429 (12.8%) subjects were diagnosed as DM, 1 405 (42.1%) were diagnosed as impaired glucose regulation (IGR) and 1 509(45.1%) had normal glucose tolerance (NGT).With a deterioration of glucose tolerance status, the difference between SB-2 hBG and OGTT-2 hBG increased gradually in 3 group (P < 0.01), namely the NGT group 1.7 (0.8-2.8) mmol/L, IGR group -0.4 (-1.2-0.6) mmol/L, DM group -2.7(-3.8-1.1) mmol/L. The cut-off points of FBG for the diagnosis of IGR and DM were 5.3 (sensitivity of 46.2%, specificity of 68.5%) and 5.6 (sensitivity of 57.4%, specificity of 76.4%) mmol/L respectively. The cut-off points of SB-2 h BG were 8.2 mmol/L for the diagnosis of IGR (sensitivity of 63.8%, specificity of 59.9%) and 9.2 mmol/L for the diagnosis of DM (sensitivity of 66.4%, specificity of 76.4%).If the cut-off point of SB-2 h BG was set at 7.2 mmol/L, the diagnostic specificity became quite low.However, at 11.1 mmol/L, the sensitivity was 31.5% and the specificity 95.7% for the diagnosis of DM. The coincidences of cut-off points of FBG and SB-2 hBG for the diagnosis of IGR and DM were equal (P > 0.05).When the cut-off point of SB-2 h BG was set at 7.8 mmol/L, the sensitivity was 77.4% and the specificity 41.8% for the diagnosis of IGR. And it was much better than FBG at 5.6 mmol/L (P < 0.01). With a deterioration of glucose tolerance, the difference between SB-2 hBG and OGTT-2 hBG increases gradually. Compared to the diagnostic criteria of OGTT, the optimal cut-off points for the diagnosis of IGR and DM were 5.3 vs 5.6 mmol/L for FBG and 8.2 vs 9.2 mmol/L for SB-2 hBG respectively.For diabetic screening in middle-aged and elders, the cut-off points of FBG at 5.3 mmol/L and SB-2 hBG at 7.8 mmol/L are indicators for further OGTT.


Folate-Mediated Targeted Delivery of siPLK1 by Leucine-Bearing Polyethylenimine.
siRNA-mediated polo-like kinase 1 (PLK1) silencing has been proposed as a promising therapeutic method for multiple cancers. However, the clinic application of this method is still hindered by the low specific delivery of siPLK1 to desired tumor lesions. Herein, folate (FA)-modified and leucine-bearing polyethylenimine was successfully synthesized and showed excellent targeted silencing to folate receptor overexpressed cells. The condensation of siPLK1 by FA-N-Ac-L-Leu-PEI (NPF) was detected by the gel retardation assay. The targeted and silencing efficiency was evaluated by flow cytometry and confocal laser scanning microscope. The PLK1 expressions at gene or protein levels were detected by quantitative real-time PCR and Western blotting assay. Further impacts of the PLK1 silencing on cell viability, cell cycle, migration, and invasion were studied by MTT, colony formation, wound healing and transwell assays. The NPF and siPLK1 could efficiently assemble to stable nanoparticles at a weight ratio of 3.0 and showed excellent condensation and protection effect. Owing to the FA-mediated targeted delivery, the uptake and silencing efficiency of NPF/siPLK1 to SGC-7901 cells was higher than that without FA modification. Moreover, NPF-mediated PLK1 silencing showed significant antitumor activity in vitro. The anti-proliferation effect of PLK1 silencing was induced via the mitochondrial-dependent apoptosis pathway with the cell cycle arrest of 45% at G2 phase and the apoptotic ratio of 28.3%. FA-N-Ac-L-Leu-PEI (NPF) could generate targeted delivery siPLK1 to FA receptor overexpressed cells and dramatically downregulate the expression of PLK1 expression.


[Cochlear function study by registration of otoacoustic emission of different classes in persons exposed to intensive industrial noise].
To investigate the condition of the acoustic analyzer and to identify early signs of affection of the internal ear receptors in persons exposed to intensive industrial noise, we examined engine-drivers exposed to noise for 1 to 20 and more years. The findings suggest that registration of otoacoustic emission of various classes may hold perspective for obtaining information about the condition of preneural micromechanical elements of the internal ear in hypoacusis of noise origin. This method detects the earliest cochlear involvement when tonic thresholds are low informative for diagnosis and prediction of hearing loss. Further study of the issue by regular audiological monitoring of persons exposed to intensive industrial noise including registration of otoacoustic emission is required.