How to use Chlamydia antibody testing in subfertility patients.
Screening for tubal factor subfertility by means of Chlamydia antibody testing (CAT) was introduced into the initial work-up of subfertile couples several years ago. The results reported, however, are heterogeneous, and no uniformity exists in cut-off levels of titres, or in definitions of tubal factor subfertility. We performed a prospective cohort study to evaluate the implications of varying the definitions of tubal pathology and of modifying the cut-off levels on the clinical impact of CAT in predicting tubal factor subfertility. In 227 consecutive patients who attended our fertility clinic, the Chlamydia IgG antibody titre was determined and related to tuboperitoneal abnormalities at laparoscopy as a reference standard. According to received operating characteristic (ROC) curve analysis, a titre of 16 is the optimum cut-off level. Increasing the cut-off level improves specificity and positive likelihood ratio (LR+), at the expense of sensitivity and negative LR (LR-). Changing the definition of tubal factor subfertility from unspecified tuboperitoneal abnormalities into extensive adhesions and/or bilateral distal tubal occlusion improves LR+, LR- and kappa significantly. We conclude that CAT is more accurate in predicting severe distal tubal pathology than unspecified tuboperitoneal abnormalities. Although from a statistical point of view a titre of 16 is the optimum cut-off level, from a clinical point of view 32 or 64 may be preferable, depending on the aim of screening and the inception cohort.


Antimicrobial resistance of fecal indicators in disinfected wastewater.
The main objective of the study was to assess the potential of three systems (UV irradiation, ozonation, and micro/ultrafiltration) operated in a pilot scale in removal of antimicrobial-resistant fecal bacteria from secondary effluent of the local wastewater treatment plant (700,000 population equivalent). The effectiveness of the processes was analysed using the removal ratio of fecal indicators (Escherichia coli and Enterococcus spp.). The susceptibility of fecal indicators to antimicrobial agents important in human therapy was examined. Resistance to nitrofurantoin and erythromycin was common among enterococci and followed by resistance to fluoroquinolones and tetracycline. Resistance to high-level aminoglycosides and glycopeptides was also observed. E. coli isolates were most frequently resistant to penicillins and tetracycline. The extended-spectrum beta-lactamase-producing E. coli was detected once, after ozonation. Substantial attention should be paid to the E. coli and enterococci resistant to three or more chemical classes of antimicrobials (MAR), which in general constituted up to 15 and 49% of the tested isolates, respectively. Although the applied methods were effective in elimination of fecal indicators (removal efficiency up to 99.99%), special attention has to be paid to the application of sufficient disinfection and operation conditions to avoid selection of antimicrobial resistant bacteria.


[Clinical significance of the procalcitonin test in differential diagnosis of systemic inflammatory reaction in pancreatonecrosis].
The significance of procalcitonin test (PCT) in the differential diagnosis of sterile and infectious pancreonecrosis was evaluated. Twenty-four patients with destructive pancreatitis were examined. Plasma procalcitonin concentrations were increased in the patients with infectious pancreonecrosis and correlated with the clinical status of patients evaluated by APACHE II and SOFA scores. The sensitivity and specificity of this test in the diagnosis of infectious pancreonecrosis were 84.4 and 83.3%, respectively, its prognostic significance being as high as 93.1%. Plasma concentrations of procalcitonin in sterile pancreonecrosis were normal or slightly increased. These results indicate that measurement of procalcitonin concentrations is an optimal laboratory test for differential diagnosis of sterile and infectious pancreonecrosis. PCT helps determine the treatment strategy in patients with various forms of destructive pancreatitis.


Pharmacokinetic interaction between rifampin and zidovudine.
A potential pharmacokinetic interaction between rifampin (Rimactan, Rifadin) and zidovudine (AZT, Retrovir) was investigated in the population of human immunodeficiency virus-infected patients at our hospital. The results from four patients who were on long-term (> or = 6 months) combination therapy with zidovudine and rifampin are presented. In all cases of combined use of zidovudine and rifampin, a lower area under the plasma concentration-time curve (AUC) and, consequently, a higher apparent clearance of zidovudine were found, compared with a reference population of zidovudine users. Patients had a low to normal maximum concentration of zidovudine in plasma. Elimination half-lives were normal in all but one patient. Zidovudine glucuronide concentrations were determined in three patients and three control subjects. The patients all had relatively higher peak plasma concentrations and higher AUCs of zidovudine glucuronide than the control subjects. In one patient, zidovudine and zidovudine glucuronide were also measured 2.5 months after discontinuation of rifampin. The AUC of zidovudine increased by a factor of 2. These data are in agreement with an enzyme-inducing effect of rifampin on the glucuronidation of zidovudine. They indicate that long-term combination therapy of rifampin and zidovudine leads to increased clearance of zidovudine, which may have therapeutic consequences.


Strategies for improving family engagement during family-centered rounds.
Family-centered rounds (FCR) are recommended as standard practice in the pediatric inpatient setting; however, limited data exist on best practices promoting family engagement during rounds. To identify strategies to enhance family engagement during FCR using a recognized systems engineering approach. In this qualitative study, stimulated recall interviews using video-recorded rounding sessions were conducted with participants representing the various stakeholders on rounds (15 parents/children and 22 healthcare team [HCT] members) from 4 inpatient services at a children's hospital in Wisconsin. On video review, participants were asked to provide strategies that would increase family engagement on FCR. Qualitative content analysis of interview transcripts was performed in an iterative process. We identified 21 categories of strategies corresponding to 2 themes related to the structure and process of FCR. Strategies related to the structure of FCR were associated with all five recognized work system elements: people (HCT composition), tasks (HCT roles), organization (scheduling of rounds and HCT training), environment (location of rounds and HCT positioning), and tools and technologies (computer use). Strategies related to the FCR process were associated with three rounding phases: before (HCT and family preparation), during (eg, introductions, presentation content, communication style), and after (follow-up) FCR. We identified a range of strategies to enhance family engagement during FCR. These strategies both confirm prior work on the importance of the content and style of communication on rounds and highlight other factors within the hospital work system, like scheduling and computer use, which may affect family engagement in care.


Effect of language on colorectal cancer screening among Latinos and non-Latinos.
Language barriers among some Latinos may contribute to the lower rates of colorectal cancer (CRC) screening between Latinos and non-Latino Whites. The purpose of this study was to examine the relationship between language and receipt of CRC screening tests among Latinos and non-Latinos using a geographically diverse, population-based sample of adults. Cross-sectional analysis of the Behavioral Risk Factor Surveillance System (BRFSS) survey. Analysis included adults age 50 years and older, who completed the 2006 BRFSS in a state that recorded data from English- and Spanish-speaking participants. The primary outcome measure was receipt of colorectal screening tests (fecal occult blood testing within prior 12 months and/or lower endoscopy within 10 years). Of the 99,895 respondents included in the study populations, 33% of Latinos responding-in-Spanish reported having had CRC testing, whereas 51% of Latinos responding-in-English and 62% of English-speaking non-Latinos reported test receipt. In multivariable analysis, compared with non-Latinos, Latinos responding-in-English were 16% less likely to have received CRC testing [odds ratio (OR), 0.84; 95% confidence interval (95% CI), 0.73-0.98], and Latinos responding-in-Spanish were 43% less likely to have received CRC testing (OR, 0.57; 95% CI, 0.44-0.74). Additionally, compared with Latinos responding-in-English, Latinos responding-in-Spanish were 36% less likely to have received CRC testing (OR, 0.64; 95% CI, 0.48-0.84). Latinos responding to the 2006 BRFSS survey in Spanish had a significantly lower likelihood of receiving CRC screening tests compared with non-Latinos and to Latinos responding-in-English. Based on this analysis, Spanish language use is negatively associated with CRC screening and may contribute to disparities in CRC screening.


Cross sectional study on cytokine production (TNF-α, IL-8) in German coalminers with progressive massive fibrosis and in control miners using a rapid wholeblood assay.
Whole-blood release of tumour necrosis factor (TNF-α) and interleukin 8 (IL-8) was studied in 26 German ex-coalminers with progressive massive fibrosis (≥A, ILO 1980; cases) and 26 ex-miners free of pneumoconiosis (≤ 0/1; controls) using a simple wholeblood assay. Cases and controls were matched individually by age and duration of coalmine dust exposure (5-year window). Whole-blood cytokine release was determined (blinded to case control status) in incubations without additions (spontaneous) and with endotoxin (LPS, 3 ng ml(-1)) or with coalmine dust (CMD; 5 mg ml(-1)). CMD-stimulated TNF-α release was significantly increased and LPS-induced IL-8 release was significantly decreased in cases (matched t-tests: p < 0.01). No effect of duration of exposure was detectable in an unmatched analysis. No clear relationship with lung function parameters independent from case/control-status was observed, although a possible positive association with central airway resistance was indicated by multiple regression for both CMD stimulated TNF-α and LPS-stimulated IL-8. This study on individually matched coalminers validates previous findings on monocyte TNF release as a marker for pneumoconiosis using a method (whole-blood assay) that is more feasible for epidemiological studies. The different response of TNF-α and IL-8 may be useful in studying the occurrence of different endpoints like pneumoconiosis and lung function decrease.


Treatment of rheumatoid arthritis with methotrexate alone, sulfasalazine and hydroxychloroquine, or a combination of all three medications.
Rheumatoid arthritis is a common disease that causes substantial morbidity and mortality. The responses of patients with rheumatoid arthritis to treatment with a single so-called disease-modifying drug, such as methotrexate, are often suboptimal. Despite limited data, many patients are treated with combinations of these drugs. We enrolled 102 patients with rheumatoid arthritis and poor responses to at least one disease-modifying drug in a two-year, double-blind, randomized study of treatment with methotrexate alone (7.5 to 17.5 mg per week), the combination of sulfasalazine (500 mg twice daily) and hydroxychloroquine (200 mg twice daily), or all three drugs. The dose of methotrexate was adjusted in an attempt to achieve remission in all patients. The primary and point of the study was the successful completion of two years of treatment with 50 percent improvement in composite symptoms of arthritis and no evidence of drug toxicity. Fifty of the 102 patients had 50 percent improvement at nine months and maintained at least that degree of improvement for two years without evidence of major drug toxicity. Among them were 24 of 31 patients treated with all three drugs (77 percent), 12 of 36 patients treated with methotrexate alone (33 percent, P < 0.001 for the comparison with the three-drug group), and 14 of 35 patients treated with sulfasalazine and hydroxychloroquine (40 percent), P = 0.003 for the comparison with the three-drug group). Seven patients in the methotrexate group and three patients in each of the other two groups discontinued treatment because of drug toxicity. In patients with rheumatoid arthritis, combination therapy with methotrexate, sulfasalazine, and hydroxychloroquine is more effective than either methotrexate alone or a combination of sulfasalazine, and hydroxychloroquine.


High Throughput Screening of FDA-Approved Drug Library Reveals the Compounds that Promote IRF3-Mediated Pro-Apoptotic Pathway Inhibit Virus Replication.
Interferon (IFN) regulatory factor 3 (IRF3) is the key transcription factor for the induction of IFN and antiviral genes. The absence of antiviral genes in IRF3 deficiency leads to susceptibility to a wide range of viral infections. Previously, we uncovered a function for nontranscriptional IRF3 (nt-IRF3), RLR (RIG-I-like receptor)-induced IRF3-mediated pathway of apoptosis (RIPA), which triggers apoptotic killing of virus-infected cells. Using knock-in mice expressing a transcriptionally inactive, but RIPA-active, IRF3 mutant, we demonstrated the relative contribution of RIPA to host antiviral defense. Given that RIPA is a cellular antiviral pathway, we hypothesized that small molecules that promote RIPA in virus-infected cells would act as antiviral agents. To test this, we conducted a high throughput screen of a library of FDA-approved drugs to identify novel RIPA activators. Our screen identified doxorubicin as a potent RIPA-activating agent. In support of our hypothesis, doxorubicin inhibited the replication of vesicular stomatitis virus, a model rhabdovirus, and its antiviral activity depended on its ability to activate IRF3 in RIPA. Surprisingly, doxorubicin inhibited the transcriptional activity of IRF3. The antiviral activity of doxorubicin was expanded to flavivirus and herpesvirus that also activate IRF3. Mechanistically, doxorubicin promoted RIPA by activating the extracellular signal-regulated kinase (ERK) signaling pathway. Finally, we validated these results using another RIPA-activating compound, pyrvinium pamoate, which showed a similar antiviral effect without affecting the transcriptional activity of IRF3. Therefore, we demonstrate that the RIPA branch of IRF3 can be targeted therapeutically to prevent virus infection.


Sensitivity of rabbit platelet and aortic cyclooxygenase to inhibition by aspirin.
It has been suggested that aspirin (ASA) would be more effective as an antithrombotic agent if employed in a dose which inhibits platelet thromboxane synthesis selectively, sparing vascular synthesis of prostaglandin I2 (PGI2). We have studied the effect of ASA concentration on rabbit platelet and aortic cyclooxygenase activity in vitro and the effect of administration of varying doses of ASA to rabbits on the cyclooxygenase activity of these tissues ex vivo. We also measured plasma levels of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) after infusion of arachidonic acid into rabbits pretreated with varying doses of ASA. Concentrations or doses of ASA required for 50% inhibition were about 10 times greater for the arterial enzyme than for the platelet enzyme in the in vitro and ex vivo studies. However, the dose required for complete inhibition of the platelet enzyme was 1 mg/kg and this dose inhibited the vascular enzyme by 62%. We conclude that meaningful selective inhibition of cyclooxygenase activity of the two tissues is difficult to achieve in the rabbit. Since doses of ASA which are antithrombotic appear to be high enough to almost totally inhibit vascular PGI2 synthesis, PGI2 synthesis may be a relatively unimportant mechanism for prevention of thrombosis.


Dynamic clustering of lipids in hydrated two-component membranes: results of computer modeling and putative biological impact.
Delineation and analysis of lateral clustering of lipids in model bilayers is an important step toward understanding of the physical processes underlying formation of lipid domains and rafts in cell membranes. Computer modeling methods represent a powerful tool to address the problem since they can detect clusters of only few lipid molecules - this issue still resists easy characterization with modern experimental techniques. In this work, we propose a computational method to detect and analyze parts of membrane with different packing densities and hydrogen bonding patterns. A series of one- and two-component fluid systems containing lipids with the same polar heads and different acyl chains, dioleoylphosphatidylcholine (18:1) and dipalmitoylphosphatidylcholine (16:0), or with same acyl chains and different polar heads, dioleoylphosphatidylserine (18:1) and dioleoylphosphatidylcholine (18:1), were studied via molecular dynamics simulations. Four criteria of clustering were considered. It was shown that the water-lipid interface of biomembranes represents a highly dynamic and "mosaic" picture, whose parameters depend on the bilayer composition. Some systems (e.g. with 20-30% of the anionic lipid) demonstrate unusual clustering properties and demand further investigation at molecular level. Lateral microheterogeneities in fluid lipid bilayers seem to be among the most important factors determining the nature of the membrane-water interface in a cell.


Cloning and characterization of Xenopus laevis drg2, a member of the developmentally regulated GTP-binding protein subfamily.
The developmentally regulated GTP-binding protein (DRG) subfamily is an uncharacterized member of the Obg family, an evolutional branch of GTPase superfamily proteins. GTPases act as molecular switches regulating diverse cellular processes. DRG2 and DRG1 comprise the DRG subfamily in eucaryotes. Although drg1 was first identified as a gene predominantly expressed during early development of the mouse central nervous system, comparative analysis of drg2 and drg1 expression during embryogenesis has never been reported, and the biochemical properties of the DRG family proteins remain to be elucidated. Thus, we first cloned Xenopus drg2 (Xdrg2) and examined the temporal and spatial expression patterns of Xdrg2 mRNA in comparison to those of Xdrg1. Both Xdrg2 and Xdrg1 are induced at late gastrula and subsequently increased during later stages of embryos (stage 13-41). Whole-mount in situ hybridization showed that Xdrg2 and Xdrg1 expression patterns are almost identical except that only Xdrg2 expression is detected in the stage 22 pronephric anlage. Strong transcripts of both genes are also observed at this stage in neural crest cells, blood islands, and developing eyes, and in brain, eyes, otic vesicle, branchial arches, pronephroses, spinal cord, notochord, head mesenchyme, and somites at stages 27 and 32. Northern blot analysis of adult tissues revealed that both genes are expressed highly in ovary and testis and rather moderately in other organs, except that Xdrg1 transcripts are scarcely detected in heart, lung, and liver. Accordingly, transcription or stability of Xdrg2 and Xdrg1 mRNAs may be regulated by different mechanisms. In addition, by generating recombinant XDRG2 and XDRG1 proteins, we found the RNA binding activity of these proteins in vitro. Our results suggest that the DRG proteins may play their physiological roles via RNA binding.


Surface-exposed silver nanoclusters inside molecular metal oxide cavities.
The surfaces of metal nanoclusters, including their interface with metal oxides, exhibit a high reactivity that is attractive for practical purposes. This high reactivity, however, has also hindered the synthesis of structurally well-defined hybrids of metal nanoclusters and metal oxides with exposed surfaces and/or interfaces. Here we report the sequential synthesis of structurally well-defined {Ag30} nanoclusters in the cavity of ring-shaped molecular metal oxides known as polyoxometalates. The {Ag30} nanoclusters possess exposed silver surfaces yet are stabilized both in solution and the solid state by the surrounding ring-shaped polyoxometalate species. The clusters underwent a redox-induced structural transformation without undesirable agglomeration or decomposition. Furthermore, {Ag30} nanoclusters showed high catalytic activity for the selective reduction of several organic functional groups using H2 under mild reaction conditions. We believe that these findings will serve for the discrete synthesis of surface-exposed metal nanoclusters stabilized by molecular metal oxides, which may in turn find applications in, for example, the fields of catalysis and energy conversion.


[The evaluation of ratio of oncomarkers in search of initial focus of tumor: on the occasion of fiftieth anniversary of discovery of alpha-fetoprotein as an oncomarker].
The article presents research data testifying the dominant value of HAFP behavior in diagnostic of oncological diseases. The importance of study of profile of main oncomarkers in patient is demonstrated. The method of hydrohelium biochips, developed in the institute of molecular biology, was used to determine 9 key oncomarkers. The application of this method made it possible to essentially complete the information map in 8 patients according to clinical interpretation of disease. In economically justified variant, this method is able to shorten period of study of patient, to specify character of pathological process and to transfer examination load of patients to the out-patient level.


ERP evidence for conflict in contingency learning.
The proportion congruency effect refers to the observation that the magnitude of the Stroop effect increases as the proportion of congruent trials in a block increases. Contemporary work shows that proportion effects can be driven by both context and individual items, and are referred to as context-specific proportion congruency (CSPC) and item-specific proportion congruency (ISPC) effects, respectively. The conflict-modulated Hebbian learning account posits that these effects manifest from the same mechanism, while the parallel episodic processing model posits that the ISPC can occur by simple associative learning. Our prior work showed that the neural correlates of the CSPC is an N2 over frontocentral electrode sites approximately 300 ms after stimulus onset that predicts behavioral performance. There is strong consensus in the field that this N2 signal is associated with conflict detection in the medial frontal cortex. The experiment reported here assesses whether the same qualitative electrophysiological pattern of results holds for the ISPC. We find that the spatial topography of the N2 is similar but slightly delayed with a peak onset of approximately 300 ms after stimulus onset. We argue that this provides strong evidence that a single common mechanism-conflict-modulated Hebbian learning-drives both the ISPC and CSPC.


Brain microemboli during cardiac surgery or aortography.
We have observed many focal dilatations or very small aneurysms in terminal arterioles and capillaries of 4 of 5 patients and 6 dogs who had recently undergone cardiopulmonary bypass. A smaller number of sausagelike dilatations distended medium-sized arterioles. Two other patients had a small number of the same microvascular changes following proximal aortography. Thirty-four patients and 6 dogs not undergoing cardiopulmonary bypass had none. (A 35th patient who had not undergone cardiopulmonary bypass or aortography showed a small number of dilatations; mediastinal air was a suggested source.) Some of the dilatations exhibited various forms of birefringence. Because most of the dilatations appear empty, we speculate that they are the sites of gas bubbles or fat emboli that have been removed by the solvents used in processing. These microvascular events, occurring only in conjunction with major arterial interventions, may be the anatomical correlate of the neurological deficits or moderate to severe intellectual dysfunction seen in at least 24% of patients after cardiac surgical procedures assisted by cardiopulmonary bypass.


Purification of human adult and foetal intestinal alkaline phosphatases by monoclonal antibody immunoaffinity chromatography.
We have used the technique of monoclonal antibody immunoaffinity chromatography to purify adult and foetal intestinal alkaline phosphatases. Pure adult intestinal enzyme was obtained from a crude tissue extract with a single immunoaffinity chromatographic step in yields exceeding 95%. An additional ion-exchange chromatographic step was necessary for purification of the foetal enzyme, but yields still exceeded 70%. Experiments to optimize the efficiency of the monoclonal antibody immunoaffinity chromatography procedure suggest that the relative strength of binding of an antibody to its antigen is the most important factor to consider when constructing such columns. A column made from an antibody of too low an avidity will not retain the enzyme, while one of too high an avidity will make elution of enzyme in the active state difficult. A scheme is suggested for the application of this technique to a general approach to enzyme purification.


Fatigue in Rheumatoid Arthritis.
The purpose of this study was to review the current information on fatigue in rheumatoid arthritis (RA). Severe fatigue is common among individuals with RA and has a significant impact on quality of life (QOL). RA-related factors (e.g., inflammation, pain) are associated with greater fatigue, but other factors, such as obesity, physical inactivity, sleep disturbance, and depression, explain the majority of variation in fatigue. Medications targeting RA have little effect on fatigue. Instead, the most effective interventions seem to address non-RA-specific factors such as physical inactivity or use cognitive behavioral approaches. No recommendations have been made for tools to measure fatigue in RA, leading to potential difficulty comparing studies. Although fatigue has great impact on patients' QOL, effective interventions that are feasible for broad dissemination remain elusive. Additional multi-faceted research is needed to identify modifiable sources of fatigue. Such research would be enhanced by harmonization of fatigue measurement across studies.


Abnormal left ventricular wall motion at mid-ejection in patients with coronary heart disease.
The degree of inward motion at mid-ejection was examined for seven segments on the silhouettes of left ventriculograms taken in the 30 degrees RAO projection in patients with normal coronary arteries. The pattern of wall motion described in these patients was used to distinguish abnormalities in mid-systolic wall motion. One or more abnormally contracting segments were found at mid-ejection of 27 of 42 patients with obstructive coronary artery disease and normal end-systolic wall motion. Of the 57 segments found in these patients, 41 or 72% corresponded to sites of significant coronary artery obstruction. Seven patients had electrocardiographic evidence of prior infarction. Following coronary graft surgery in eight patients improved motion was found in association with graft patency in seven priviously delayed segments and two new areas of delayed wall motion associated with nonpatent grafts and electrocardiographic changes of infarction appeared. We postulate that some of the myocardial fibers in late contracting segments have been injured or infarcted and are able to contract effectively only during the latter half of ejection when ventricular wall tension is reduced.


p21(WAF1) is component of a positive feedback loop that maintains the p53 transcriptional program.
The regulation of p53 activity through the MDM2 negative feedback loop is driven in part by an extrinsic ATM-pulse that maintains p53 oscillations in response to DNA damage. We report here that the p53 pathway has evolved an intrinsic positive feedback loop that is maintained by the p53-inducible gene product p21(WAF1). p21-null cancer cells have defects in p53 protein turnover, reductions in MDM2-mediated degradation of p53, and reduced DNA damage-induced ubiquitination of p53. TLR3-IRF1 or ATM-dependent signaling to p53 is defective in p21-null cells and complementation of the p21 gene in p21-null cancer cells restores the p53 transcriptional response. The mechanism of p53 inactivity in p21-null cells is linked to a p53 protein equilibrium shift from chromatin into cytosolic fractions and complementation of the p21 gene into p21-null cells restores the nuclear localization of p53. A loss of p53 transcriptional function in murine B-cells heterozygous or homozygous null for p21 highlights a p21-gene dosage effect that maintains the full p53 transcriptional response. ATM inhibition results in nuclear exclusion of p53 highlighting a positive genetic interaction between ATM and p21. P21 protein oscillates in undamaged proliferating cells, and reductions of p21 protein using siRNA eliminate the DNA damage-induced p53 pulse. The p53 transcription program has evolved a negative feedback loop maintained by MDM2 that is counteracted by a positive feedback loop maintained by ATM-p21 the balance of which controls the specific activity of p53 as a transcription factor.


Childhood-onset systemic lupus erythematosus: a comparative study of African Americans and Latin Americans.
This study compared the clinical and serologic features in two different ethnic groups of patients with childhood-onset systemic lupus erythematosus (SLE). One hundred seventy-one SLE patients comprised the study population; 61 (55 girls and 6 boys) were African American with age at onset of 13 +/- 2.9 years, and 110 (97 girls and 13 boys) were Latin American (Colombian) with age at onset of 13 +/- 3.2 years. Clinical, demographic, and laboratory data were obtained by chart review using a standard data collection form. African-American patients more commonly manifested discoid skin lesions, malar rash, pulmonary fibrosis, and pleuritis, and less commonly manifested photosensitivity, livedo reticularis, and vascular thrombosis than did Latin Americans. In addition, there was a higher frequency of anti-dsDNA, anti-Sm, anti-RNP, and anti-Ro positivity among African-Americans compared with Latin-American patients. These results suggest the presence of ethnic differences in the clinical expression of SLE.


The global burden of respiratory infections in indigenous children and adults: A review.
This review article focuses on common lower respiratory infections (LRIs) in indigenous populations in both developed and developing countries, where data is available. Indigenous populations across the world share some commonalities including poorer health and socio-economic disadvantage compared with their non-indigenous counterparts. Generally, acute and chronic respiratory infections are more frequent and more severe in both indigenous children and adults, often resulting in substantial consequences including higher rates of bronchiectasis and poorer outcomes for patients with chronic obstructive pulmonary disease (COPD). Risk factors for the development of respiratory infections require recognition and action. These risk factors include but are not limited to socio-economic factors (e.g. education, household crowding and nutrition), environmental factors (e.g. smoke exposure and poor access to health care) and biological factors. Risk mitigation strategies should be delivered in a culturally appropriate manner and targeted to educate both individuals and communities at risk. Improving the morbidity and mortality of respiratory infections in indigenous people requires provision of best practice care and awareness of the scope of the problem by healthcare practitioners, governing bodies and policy makers.


Investigation of photoplethysmographic changes using a static compression model of spinal cord injury.
Little is known about cell death in spinal cord tissue following compression injury, despite compression being a key component of spinal injuries. Currently models are used to mimic compression injury in animals and the effects of the compression evaluated by observing the extent and duration of recovery of normal motor function in the days and weeks following the injury. A fiber-optic photoplethysmography system was used to investigate whether pulsation of the small arteries in the spinal cord occurred before, during and after compressive loads were applied to the tissue. It was found that the signal amplitudes were reduced and this reduction persisted for at least five minutes after the compression ceased. It is hoped that results from this preliminary study may improve knowledge of the mechanism of spinal cord injury.


The evolution of the G matrix: selection or drift?
The evolution of quantitative characters can be described by the equation Deltaz=GP-1S where Deltaz is the vector of mean responses, G is the matrix of additive genetic variances and covariances, P is the matrix of phenotypic variances and covariances and S is the vector of selection differentials. This equation can be used to predict changes in trait values or to retrospectively estimate the selection gradient and is thus a central equation of evolutionary quantitative genetics. Genetic variances and covariances will be reduced by stabilizing selection, directional selection and drift, and increased by mutation. Changes in trait values resulting from directional selection that are comparable with differences observed among species are readily obtainable in short geological time spans (<5000 generations) with selection intensities so small that they would have an insignificant effect on the G matrix (of course it is possible that such changes came about by strong selection over a few generations, followed by long periods of stasis; there is insufficient evidence to presently distinguish these two possibilities). On the other hand, observed effective population sizes are sufficiently small that considerable changes in G can be expected from drift alone. The action of drift can be distinguished from selection because the former produces a proportional change in G whereas the latter, in general, will not. A survey of studies examining variation in G suggests that the null hypothesis that most of the variation can be attributed to drift rather than selection cannot be rejected. However, more research on the predicted statistical distribution of G as a result of selection and/or drift is required and further development of statistical tests to distinguish these two forces needs to be made.


Yeast (1 → 3)-(1 → 6)-β-d-glucan alleviates immunosuppression in gemcitabine-treated mice.
Gemcitabine (2'-deoxy-2',2'-difluorocytidine, dFdC) is one of the most effective chemotherapy drugs commonly used for treatment of various tumors. Despite its significant anticancer effects, some adverse effects create obstacles to treatment. The main toxicity of gemcitabine is myelosuppression, which not only reduces patient quality of life, but also hinders further anticancer treatment. In this respect, immunotherapy can address these drawbacks because of its ability to enhance the patient's immune system. To improve immune system function, yeast-derived β-glucans, which are well-known biologic response modifiers, were administered to gemcitabine-treated mice. The in vivo experiment revealed that orally administered yeast (1 → 3)-(1 → 6)-β-d-glucan effectively alleviated myelosuppression associated with gemcitabine-induced pancytopenia. Moreover, analysis of myelopoiesis-related cytokine expression through real-time PCR demonstrated that β-glucan treatment significantly upregulated hematopoietic responses in gemcitabine-treated mice. Furthermore, orally administered β-glucan significantly induced the expression of IFN-γ and IL-2 in splenocytes of gemcitabine-treated mice. It also restored the cytotoxicity of splenocytes against YAC-1 in gemcitabine-treated mice and displayed a positive effect on gemcitabine-damaged bone marrow tissue. In conclusion, yeast β-glucans have the potential to be used as adjuvants for alleviating chemotherapy-induced immunosuppression in patients.


Investigating the association between obstructive sleep apnea and periodontitis.
Obstructive sleep apnea (OSA) is a sleep disorder characterized by disruptions of normal sleep architecture. Chronic periodontitis is a chronic disease of the periodontium that elicits a general inflammatory response to local dental plaque. It has been suggested that periodontal disease may increase in severity with increasingly severe OSA because both disease entities share common inflammatory pathways, acting synergistically to alter the host response. The aim of this study is to analyze the association between severity of OSA and the prevalence/severity of periodontitis. One hundred patients from a large veterans administration sleep study center (n = 26 normal, n = 21 mild, n = 19 moderate, n = 34 severe) diagnosed with an overnight polysomnogram underwent a comprehensive periodontal examination. Periodontal parameters measured included the following: 1) mean periodontal probing depth (PD); 2) clinical attachment level (CAL); 3) gingival recession; and 4) percentage of sites with bleeding on probing, plaque, PD ≥5 mm, and CAL ≥3 mm. Seventy-three percent of the sampled population had moderate/severe periodontal disease. χ(2) analyses revealed no significant differences in the prevalence of periodontal disease between the apnea-hypopnea index (AHI) groups, with a negligible Spearman correlation coefficient of 0.246 between AHI severity and periodontal disease severity categories. Analysis of covariance indicated a significant association between AHI severity categories and percentage of sites with plaque, after adjusting for age. Multivariable logistic regression analysis predicting moderate/severe periodontitis with AHI score, age, and smoking status indicated a significant association with age (P = 0.028) but no significant association with the other two predictors. OSA was not significantly associated with the prevalence of moderate/severe periodontitis and the periodontal parameters examined, except percentage plaque.


Photobiomodulation therapy reduces apoptotic factors and increases glutathione levels in a neuropathic pain model.
Neuropathic pain (NP) is caused by damage to the nervous system due to reactive oxygen spices (ROS) increase, antioxidants reduction, ATP production imbalance, and induction of apoptosis. In this investigation, we applied low-level laser 660 nm (photobiomodulation therapy) as a new strategy to modulate pain. In order to study the effects of photobiomodulation therapy (660 nm) on NP, chronic constriction injury (CCI) model was selected. Low-level laser of 660 nm was used for 2 weeks. Thermal and mechanical hyperalgesia were measured before and after surgery on days 7 and 14, respectively. Paw withdrawal thresholds were also evaluated. Expression of p2x3, Bax, and bcl2 protein was measured by western blotting. The amount of glutathione (GSH) was measured in the spinal cord by continuous spectrophotometric rate determination method. The results are presented as mean ± SD. Statistical analysis of data was carried out using SPSS 21. CCI decreased the pain threshold, 2-week photobiomodulation therapy significantly increased mechanical and thermal threshold, decreased P2X3 expression (p < 0.001), and increased bcl2 expression (p < 0.01), but it was not effective on the Bax expression. We speculated that although photobiomodulation therapy increased ROS generation, it increased antioxidants such as GSH. Increase in bcl2 is another mitochondrial protection mechanism for cell survival and that pain relief and decrease in P2X3 expression confirm it.


The relationship between sales of SSRI, TCA and suicide rates in the Nordic countries.
In the period 1990-2006, strong and almost equivalent increases in sales figures of selective serotonin re-uptake inhibitors (SSRIs) were observed in all Nordic countries. The sales figures of tricyclic antidepressants (TCAs) dropped in Norway and Sweden in the nineties. After 2000, sales figures of TCAs have been almost constant in all Nordic countries. The potentially toxic effect of TCAs in overdose was an important reason for replacing TCAs with SSRIs when treating depression. We studied whether the rapid increase in sales of SSRIs and the corresponding decline in TCAs in the period 1990-98 were associated with a decline in suicide rates. Aggregated suicide rates for the period 1975-2006 in four Nordic countries (Denmark, Finland, Norway and Sweden) were obtained from the national causes-of-death registries. The sales figures of antidepressants were provided from the wholesale registers in each of the Nordic countries. Data were analysed using Fisher's exact test and Pearson's correlation coefficient. There was no statistical association (P = 1.0) between the increase of sales figures of SSRIs and the decline in suicide rates. There was no statistical association (P = 1.0) between the decrease in the sale figures of TCAs and change in suicide rates either. We found no evidence for the rapid increase in use of SSRIs and the corresponding decline in sales of TCAs being associated with a decline in the suicide rates in the Nordic countries in the period 1990-98. We did not find any inverse relationship between the increase in sales of SSRIs and declining suicide rates in four Nordic countries.


Regulation of protein-protein binding by coupling between phosphorylation and intrinsic disorder: analysis of human protein complexes.
Phosphorylation offers a dynamic way to regulate protein activity, subcellular localization, and stability. The majority of signaling pathways involve an extensive set of protein-protein interactions, and phosphorylation is widely used to regulate protein-protein binding by affecting the stability, kinetics and specificity of interactions. Previously it was found that phosphorylation sites tend to be located on protein-protein binding interfaces and may orthosterically modulate the strength of interactions. Here we studied the effect of phosphorylation on protein binding in relation to intrinsic disorder for different types of human protein complexes with known structure of the binding interface. Our results suggest that the processes of phosphorylation, binding and disorder-order transitions are coupled to each other, with about one quarter of all disordered interface Ser/Thr/Tyr sites being phosphorylated. Namely, residue site disorder and interfacial states significantly affect the phosphorylation of serine and to a lesser extent of threonine. Tyrosine phosphorylation might not be directly associated with binding through disorder, and is often observed in ordered interface regions which are not predicted to be disordered in the unbound state. We analyze possible mechanisms of how phosphorylation might regulate protein-protein binding via intrinsic disorder, and specifically focus on how phosphorylation could prevent disorder-order transitions upon binding.


Beef customer satisfaction: USDA quality grade and marination effects on consumer evaluations of top round steaks.
An in-home beef study evaluated consumer ratings of top round steaks (semimembranosus) as influenced by USDA quality grade (top Choice or high Select), city (Chicago or Philadelphia), consumer segment (beef loyalists = heavy consumers of beef; budget rotators = cost-driven and split meat consumption between beef and chicken; and variety rotators = higher incomes and education and split meat consumption among beef, poultry, and other foods), degree of doneness, cooking method, and marination. Consumers evaluated each steak for overall like, tenderness, juiciness, flavor like, and flavor amount using 10-point scales (1 = dislike extremely, not at all tender, not at all juicy, dislike extremely, and none at all to 10 = like extremely, extremely tender, extremely juicy, like extremely, and an extreme amount of flavor, respectively). Quality grade affected several consumer sensory traits, with top Choice receiving higher (P < or = 0.004) tenderness, juiciness, and flavor like scores than high Select. Consumers in Chicago rated steaks cooked "medium and less" higher for overall like, tenderness, juiciness, flavor like, and flavor amount than those in Philadelphia (city x degree of doneness; P < or = 0.020). Steaks braised by customers in Philadelphia received among the highest scores for overall like, tenderness, juiciness, flavor like, and flavor amount compared with any cooking method used by customers in Chicago (cooking method x city; P < or = 0.026). Overall like and flavor amount ratings were least (P < 0.05) for steaks that were marinated and cooked to "medium and less" degree of doneness (marination x degree of doneness; P < or = 0.014). Braised steaks received among the highest values for overall like, tenderness, juiciness, flavor like, and flavor amount when cooked to "medium and less" or "medium well and more" (cooking method x degree of doneness; P < or = 0.008). Correlation and stepwise regression analysis indicated that flavor like was pivotal in customers' satisfaction with top round steaks, and was the sensory trait most highly correlated to overall like, followed by tenderness, flavor amount, and juiciness. Preparation of top round steaks was crucial in consumers' likes and dislikes, and by improving flavor, higher consumer satisfaction may be achieved.


Age-dependent change of coalitionary strategy in male Barbary macaques.
Inter- and intra-specific variation in the propensity to form coalitions has been explained by variation in the availability of suitable partners, distribution of fighting ability, coalition profitability, and costs of coordination. Male coalition formation can be an alternative reproductive strategy to one-on-one competition to maximize male reproductive success. Here we focus on age as a state variable to explain within-group variation in individual propensity to form coalitions against other group members. We specifically test the prediction that males conditionally switch from a solo strategy for achievement of high mating success to a cooperative strategy after reaching post-prime age in male Barbary macaques (Macaca sylvanus). We combined new observations with data collected in 2006 and 2008 on the same individuals from one captive group living in semi-natural conditions at Affenberg Salem, Germany, and found that in all years males between 5 and 13 years formed significantly fewer coalitions than males 14 years and older (post-prime). More importantly, we found those males that aged into the post-prime phase to have switched their reproductive strategy and to form significantly more coalitions in 2014 compared to 2008. These first longitudinal data together with earlier cross-sectional analyses in this and other primate species suggest that group-level measures of coalition propensity may be strongly affected by the age composition of groups and that male coalition formation can be a conditional reproductive strategy.


Life-Saving Threads: Advances in Textile-Based Analytical Devices.
Novel approaches that incorporate electrofluidic and microfluidic technologies are reviewed to illustrate the translation of traditional enclosed structures into open and accessible textile based platforms. Through the utilization of on-fiber and on-textile microfluidics, it is possible to invert the typical enclosed capillary column or microfluidic "chip" platform, to achieve surface accessible efficient separations and fluid handling, while maintaining a microfluidic environment. The open fiber/textile based fluidics approach immediately provides new possibilities to interrogate, manipulate, redirect, extract, characterize, and quantify solutes and target species at any point in time during such processes as on-fiber electrodriven separations. This approach is revolutionary in its simplicity and provides many potential advantages not otherwise afforded by the more traditional enclosed platforms.


An automated field phenotyping pipeline for application in grapevine research.
Due to its perennial nature and size, the acquisition of phenotypic data in grapevine research is almost exclusively restricted to the field and done by visual estimation. This kind of evaluation procedure is limited by time, cost and the subjectivity of records. As a consequence, objectivity, automation and more precision of phenotypic data evaluation are needed to increase the number of samples, manage grapevine repositories, enable genetic research of new phenotypic traits and, therefore, increase the efficiency in plant research. In the present study, an automated field phenotyping pipeline was setup and applied in a plot of genetic resources. The application of the PHENObot allows image acquisition from at least 250 individual grapevines per hour directly in the field without user interaction. Data management is handled by a database (IMAGEdata). The automatic image analysis tool BIVcolor (Berries in Vineyards-color) permitted the collection of precise phenotypic data of two important fruit traits, berry size and color, within a large set of plants. The application of the PHENObot represents an automated tool for high-throughput sampling of image data in the field. The automated analysis of these images facilitates the generation of objective and precise phenotypic data on a larger scale.


Modulation of membrane curvature by phosphatidic acid and lysophosphatidic acid.
The local generation of phosphatidic acid plays a key role in the regulation of intracellular membrane transport through mechanisms which are largely unknown. Phosphatidic acid may recruit and activate downstream effectors, or change the biophysical properties of the membrane and directly induce membrane bending and/or destabilization. To evaluate these possibilities, we determined the phase properties of phosphatidic acid and lysophosphatidic acid at physiological conditions of pH and ion concentrations. In single-lipid systems, unsaturated phosphatidic acid behaved as a cylindrical, bilayer-preferring lipid at cytosolic conditions (37 degrees C, pH 7.2, 0.5 mM free Mg2+), but acquired a type-II shape at typical intra-Golgi conditions, a mildly acidic pH and submillimolar free Ca2+ (pH 6.6-5.9, 0.3 mM Ca2+). Lysophosphatidic acid formed type-I lipid micelles in the absence of divalent cations, but anhydrous cation-lysophosphatidic acid bilayer complexes in their presence. These data suggest a similar molecular shape for phosphatidic acid and lysophosphatidic acid at cytosolic conditions; however, experiments in mixed-lipid systems indicate that their shape is not identical. Lysophosphatidic acid stabilized the bilayer phase of unsaturated phosphatidylethanolamine, while the opposite effect was observed in the presence of phosphatidic acid. These results support the hypothesis that a conversion of lysophosphatidic acid into phosphatidic acid by endophilin or BARS (50 kDa brefeldin A ribosylated substrate) may induce negative spontaneous monolayer curvature and regulate endocytic and Golgi membrane fission. Alternative models for the regulation of membrane fission based on the strong dependence of the molecular shape of (lyso)phosphatidic acid on pH and divalent cations are also discussed.


EMC10 (Endoplasmic Reticulum Membrane Protein Complex Subunit 10) Is a Bone Marrow-Derived Angiogenic Growth Factor Promoting Tissue Repair After Myocardial Infarction.
Clinical trials of bone marrow cell-based therapies after acute myocardial infarction (MI) have produced mostly neutral results. Treatment with specific bone marrow cell-derived secreted proteins may provide an alternative biological approach to improving tissue repair and heart function after MI. We recently performed a bioinformatic secretome analysis in bone marrow cells from patients with acute MI and discovered a poorly characterized secreted protein, EMC10 (endoplasmic reticulum membrane protein complex subunit 10), showing activity in an angiogenic screen. We investigated the angiogenic potential of EMC10 and its mouse homolog (Emc10) in cultured endothelial cells and infarcted heart explants. We defined the cellular sources and function of Emc10 after MI using wild-type, Emc10-deficient, and Emc10 bone marrow-chimeric mice subjected to transient coronary artery ligation. Furthermore, we explored the therapeutic potential of recombinant Emc10 delivered by osmotic minipumps after MI in heart failure-prone FVB/N mice. Emc10 signaled through small GTPases, p21-activated kinase, and the p38 mitogen-activated protein kinase (MAPK)-MAPK-activated protein kinase 2 (MK2) pathway to promote actin polymerization and endothelial cell migration. Confirming the importance of these signaling events in the context of acute MI, Emc10 stimulated endothelial cell outgrowth from infarcted mouse heart explants via p38 MAPK-MK2. Emc10 protein abundance was increased in the infarcted region of the left ventricle and in the circulation of wild-type mice after MI. Emc10 expression was also increased in left ventricular tissue samples from patients with acute MI. Bone marrow-derived monocytes and macrophages were the predominant sources of Emc10 in the infarcted murine heart. Emc10 KO mice showed no cardiovascular phenotype at baseline. After MI, however, capillarization of the infarct border zone was impaired in KO mice, and the animals developed larger infarct scars and more pronounced left ventricular remodeling compared with wild-type mice. Transplanting KO mice with wild-type bone marrow cells rescued the angiogenic defect and ameliorated left ventricular remodeling. Treating FVB/N mice with recombinant Emc10 enhanced infarct border-zone capillarization and exerted a sustained beneficial effect on left ventricular remodeling. We have identified Emc10 as a previously unknown angiogenic growth factor that is produced by bone marrow-derived monocytes and macrophages as part of an endogenous adaptive response that can be enhanced therapeutically to repair the heart after MI.


Testosterone depresses innate and acquired resistance to ticks in natural rodent hosts: a force for aggregated distributions of parasites.
The effects of testosterone on acquired resistance to ticks, Ixodes ricinus, in their natural rodent hosts (voles, Clethrionomys glareolus, and wood-mice, Apodemus sylvaticus) were investigated by manipulating testosterone levels and exposing the hosts to repeated tick infestations. Testosterone reduced both innate and acquired resistance to tick feeding. During primary infestations, attachment rates were higher on rodents with high testosterone levels than on oil-implanted controls. Successive infestations on voles were accompanied by a decrease in tick feeding success and survival, but this decrease was significantly greater in ticks fed on control voles than in those fed on voles implanted with testosterone. When reduced feeding success had been induced, either by vaccination with tick salivary gland extract or by 4 successive infestations, implantation with testosterone partially reversed the acquired resistance. These effects of testosterone will generate heterogeneities within the rodent population with respect to tick distribution and microparasite transmission. The lowest innate and acquired resistance to tick feeding occurs in that fraction of the host population, i.e., sexually active males, most actively involved in the transmission of both Babesia microti and Borrelia burgdorferi s.l.


Conformational dissection of a viral intrinsically disordered domain involved in cellular transformation.
Intrinsic disorder is abundant in viral genomes and provides conformational plasticity to its protein products. In order to gain insight into its structure-function relationships, we carried out a comprehensive analysis of structural propensities within the intrinsically disordered N-terminal domain from the human papillomavirus type-16 E7 oncoprotein (E7N). Two E7N segments located within the conserved CR1 and CR2 regions present transient α-helix structure. The helix in the CR1 region spans residues L8 to L13 and overlaps with the E2F mimic linear motif. The second helix, located within the highly acidic CR2 region, presents a pH-dependent structural transition. At neutral pH the helix spans residues P17 to N29, which include the retinoblastoma tumor suppressor LxCxE binding motif (residues 21-29), while the acidic CKII-PEST region spanning residues E33 to I38 populates polyproline type II (PII) structure. At pH 5.0, the CR2 helix propagates up to residue I38 at the expense of loss of PII due to charge neutralization of acidic residues. Using truncated forms of HPV-16 E7, we confirmed that pH-induced changes in α-helix content are governed by the intrinsically disordered E7N domain. Interestingly, while at both pH the region encompassing the LxCxE motif adopts α-helical structure, the isolated 21-29 fragment including this stretch is unable to populate an α-helix even at high TFE concentrations. Thus, the E7N domain can populate dynamic but discrete structural ensembles by sampling α-helix-coil-PII-ß-sheet structures. This high plasticity may modulate the exposure of linear binding motifs responsible for its multi-target binding properties, leading to interference with key cell signaling pathways and eventually to cellular transformation by the virus.


Socioeconomic inequalities in completion of hepatitis B vaccine series among Korean women: results from a nationwide interview survey.
Hepatitis B virus (HBV) infection and its sequelae are major global health problems. This study was conducted to investigate the association between factors related to socioeconomic status and HBV vaccination in the general population of women in Korea. Data from annual nationwide cross-sectional interview surveys conducted between 2005 and 2008 were reviewed. These surveys included representative samples of cancer-free people and 4350 women interviewed within the study period were included in the present report. Polychotomous logistic regression was used in the analysis. The complete HBV coverage rate was 39.8%. Older age, lower household income and lower education level showed significant unfavorable influences on the completion of the HBV vaccine series (P-trend <0.001 for all three variables). Lack of private insurance and having a routine and manual job were also associated with a lower complete vaccination rate (OR: 0.69, 95% CI: 0.57-0.84; OR: 0.56, 95% CI: 0.38-0.82, respectively), whereas living in an urban area was related to a higher rate (OR: 1.18, 95% CI: 1.02-1.37). Older age, lower education level, and not having private insurance were associated with incomplete vaccination compared with unvaccination (P-trend 0.001, <0.001, OR: 0.68, 95% CI: 0.53-0.78, respectively). Among women of reproductive age, similar results were seen for education level, household income, and occupation. Significant disparities in vaccination status according to socioeconomic status, as indicated by the combination of household income and education level, were seen (P-trend <0.001). The most common reason for unvaccination was lack of knowledge about the necessity for HBV vaccination. Considering that vertical transmission is the most common source of HBV infection, vaccination of women is important to prevent HBV transmission to newborn infants. Therefore governmental strategies to reduce socioeconomic inequalities related to the completion of the HBV vaccine series among women should be implemented.


Wound-induced expression of a tobacco peroxidase is not enhanced by ethephon and suppressed by methyl jasmonate and coronatine.
In tobacco plants, wounding induces production of a set of defense-related proteins such as basic pathogenesis-related (PR) proteins and proteinase inhibitors (PIs) via the jasmonate/ethylene pathway. Although class III plant peroxidase (POX) is also wound-inducible, the regulatory mechanism for its wound-induced expression is not fully understood. Here, we describe that a tobacco POX gene (tpoxN1), which is constitutively expressed in roots, is induced locally 30 min after wounding and then systemically in tobacco plants. Infection of necrotizing virus also induced tpoxN1 gene. The wound-induced expression was not enhanced by known wound-signal compounds such as methyl jasmonate (MeJA) and ethephon in contrast to other wound-inducible genes such as basic PR-1 and PI-II genes. And treatment with MeJA and coronatine, biological analogs of jasmonate, rather suppressed the tpoxN1 expression. Salicylic acid, an antagonist of jasmonate-based wound signaling, did not suppress the wound-induced expression of tpoxN1. Only spermine, which is reported as an endogenous inducer for acidic PR genes in tobacco mosaic virus-infected tobacco leaves, could induce tpoxN1 gene expression. These results suggest that wound-induced expression of the tpoxN1 gene is regulated differently from that of the basic PR and PI-II genes.


Assessing recall bias and measurement error in high-frequency social data collection for human-environment research.
A major impediment to understanding human-environment interactions is that data on social systems are not collected in a way that is easily comparable to natural systems data. While many environmental variables are collected with high frequency, gridded in time and space, social data is typically conducted irregularly, in waves that are far apart in time. These efforts typically engage respondents for hours at a time, and suffer from decay in participants' ability to recall their experiences over long periods of time. Systematic use of mobile and smartphones has the potential to transcend these challenges, with a critical first step being an evaluation of where survey respondents experience the greatest recall decay. We present results from, to our knowledge, the first systematic evaluation of recall bias in components of a household survey, using the Open Data Kit (ODK) platform on Android smartphones. We tasked approximately 500 farmers in rural Bangladesh with responding regularly to components of a large household survey, randomizing the frequency of each task to be received weekly, monthly, or seasonally. We find respondents' recall of consumption and experience (such as sick days) to suffer much more greatly than their recall of the use of their households' time for labor and farm activities. Further, we demonstrate a feasible and cost-effective means of engaging respondents in rural areas to create and maintain a true socio-economic "baseline" to mirror similar efforts in the natural sciences.


Genomic organization, rapid evolution and meiotic instability of nucleotide-binding-site-encoding genes in a new fruit crop, "chestnut rose".
From chestnut rose, a promising fruit crop of the Rosa genus, powdery mildew disease-resistant and susceptible genotypes and their F(1) progeny were used to isolate nucleotide-binding-site (NBS)-encoding genes using 19 degenerate primer pairs and an additional cloning method called overlapping extension amplification. A total of 126 genes were harvested; of these, 38 were from a resistant parent, 37 from a susceptible parent, and 51 from F(1) progeny. A phylogenetic tree was constructed, which revealed that NBS sequences from parents and F(1) progeny tend to form a mixture and are well distributed among the branches of the tree. Mapping of these NBS genes suggested that their organization in the genome is a "tandem duplicated cluster" and, to a lesser extent, a "heterogeneous cluster." Intraspecific polymorphisms and interspecific divergence were detected by Southern blotting with NBS-encoding genes as probes. Sequencing on the nucleotide level revealed even more intraspecific variation: for the R4 gene, 9.81% of the nucleotides are polymorphic. Amino acid sites under positive selection were detected in the NBS region. Some NBS-encoding genes were meiotically unstable, which may due to recombination and deletion events. Moreover, a transposon-like element was isolated in the flanking region of NBS genes, implying a possible role for transposon in the evolutionary history of resistance genes.


An autoantibody to single-stranded DNA: comparison of the three-dimensional structures of the unliganded Fab and a deoxynucleotide-Fab complex.
Crystal structures of the Fabs from an autoantibody (BV04-01) with specificity for single-stranded DNA have been determined in the presence and absence of a trinucleotide of deoxythymidylic acid, d(pT)3. Formation of the ligand-protein complex was accompanied by small adjustments in the orientations of the variable (VL and VH) domains. In addition, there were local conformational changes in the first hypervariable loop of the light chain and the third hypervariable loop of the heavy chain, which together with the domain shifts led to an improvement in the complementarity of nucleotide and Fab. The sugar-phosphate chain adopted an extended and "open" conformation, with the base, sugar, and phosphate components available for interactions with the protein. Nucleotide 1 (5'-end) was associated exclusively with the heavy chain, nucleotide 2 was shared by both heavy and light chains, and nucleotide 3 was bound by the light chain. The orientation of phosphate 1 was stabilized by hydrogen bonds with serine H52a and asparagine H53. Phosphate 2 formed an ion pair with arginine H52, but no other charge-charge interactions were observed. Insertion of the side chain of histidine L27d between nucleotides 2 and 3 resulted in a bend in the sugar-phosphate chain. The most dominant contacts with the protein involved the central thymine base, which was immobilized by cooperative stacking and hydrogen bonding interactions. This base was intercalated between a tryptophan ring (no. H100a) from the heavy chain and a tyrosine ring (no. L32) from the light chain. The resulting orientation of thymine was favorable for the simultaneous formation of two hydrogen bonds with the backbone carbonyl oxygen and the side chain hydroxyl group of serine L91 (the thymine atoms were the hydrogen on nitrogen 3 and keto oxygen 4).


p34cdc2 phosphorylation sites in histone H1 are dephosphorylated by protein phosphatase 2A1.
The protein phosphatase activity in rat liver cytosol or nuclear extracts that dephosphorylates histone H1 which has been phosphorylated by p34cdc2 is inhibited completely by okadaic acid, but unaffected by inhibitor-2 or magnesium ions, demonstrating that the only enzyme in this tissue capable of dephosphorylating this substrate is a type 2A phosphatase. Fractionation of the cytosol by anion-exchange chromatography and gel filtration demonstrated that histone H1 phosphatase activity coeluted with the major species of protein phosphatase 2A, termed PP2A1 and PP2A2. PP2A1 was the most active histone H1 phosphatase, its histone phosphatase phosphorylase phosphatase activity ratio being 6-fold higher than PP2A2 and 30-fold higher than the free catalytic subunit PP2AC. It is concluded that PP2A1 is likely to be the enzyme which dephosphorylates p34cdc2-labelled histone H1 in vivo and that the A and B subunits which interact with PP2AC in this species each play a key role in facilitating dephosphorylation of this substrate. The results demonstrate that PP2A, in addition to being involved in suppressing the activation of p34cdc2 in vivo, can also function to reverse at least one of its actions.


Dynamic structure of locomotor behavior in walking fruit flies.
The function of the brain is unlikely to be understood without an accurate description of its output, yet the nature of movement elements and their organization remains an open problem. Here, movement elements are identified from dynamics of walking in flies, using unbiased criteria. On one time scale, dynamics of walking are consistent over hundreds of milliseconds, allowing elementary features to be defined. Over longer periods, walking is well described by a stochastic process composed of these elementary features, and a generative model of this process reproduces individual behavior sequences accurately over seconds or longer. Within elementary features, velocities diverge, suggesting that dynamical stability of movement elements is a weak behavioral constraint. Rather, long-term instability can be limited by the finite memory between these elementary features. This structure suggests how complex dynamics may arise in biological systems from elements whose combination need not be tuned for dynamic stability.


Early or delayed reconstruction in multi-ligament knee injuries: A systematic review and meta-analysis.
Whether early or delayed surgical intervention in patients with multi-ligament injuries results in better outcomes, is of current and controversial debate. The purpose of this study was to perform a meta-analysis comparing early versus late surgical treatment of multi-ligament knee injuries. We performed a systematic review of Medline, Embase, Scopus, and Google Scholar to identify relevant studies in the English and German literature. Eligibility criteria included studies comparing early or delayed surgical interventions for multi-ligament knee injuries, with a minimum follow-up of two years, reporting the primary clinical outcome using a validated functional scoring system and range of motion. Exclusion criteria were patients treated with multi-trauma, head injury, non-union, lower extremity fractures, or a documented history of previous knee injuries. Publication bias was assessed by funnel plot, and the risk of bias was established using the Cochrane Collaboration's risk of bias tool. Heterogeneity was assessed using χ2 and I2 statistic. Eight studies (n=260 patients) were included in the analysis. 149 patients were treated early with a mean of 10.6days; 111 patients were treated late with a mean of 294days. The pooled estimate for clinical outcome demonstrated that early surgery resulted in significantly higher Lysholm scores (SMD 0.669, 95% CI: 0.379 to 0.959, p=0.0001, I2=0%). Thirty-one per cent of all patients with early surgery had a normal or near normal knee, whereas only 15% of patients with late reconstruction reported the knee to be normal or near normal. The pooled estimates for total ROM did not demonstrate a significant difference between the groups (SMD 0.113, 95% CI: -0.271 to 0.498, p=0.564, I2=35.57%). The results of this meta-analysis suggest that early surgical intervention in multi-ligament injuries of the knee produces a significantly superior clinical outcome, compared to late reconstruction. Although an overall trend of improved total range of knee motion was also demonstrated, this was very small and unlikely to be clinically relevant. Level 4; Systematic review and meta-analysis.


Analysis of tobacco-specific nitrosamines in Moldovan cigarette tobacco.
Tobacco-specific nitrosamines (TSNA) are among the most important and abundant strongly carcinogenic agents in unburned tobacco. It has been established that 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) induces lung tumors in rodents independent of the route of administration. N'-Nitrosonornicotine (NNN) causes tumors of the esophagus and nasal cavity in rats, lung in mice, and respiratory tract in hamsters. Although the manufacturing of cigarettes is an important domain of Moldovan industry, there are no reports in the literature on TSNA analysis in Moldovan tobacco. The main purpose of the present study was an initial evaluation of TSNA levels in Moldovan cigarette tobacco. Eighteen brands of Moldovan cigarettes, representing 78% of all brands produced in Moldova, were analyzed. Four TSNA-NNN, NNK, N'-nitrosoanatabine (NAT), and N'-nitrosoanabasine (NAB)-were analyzed by gas chromatography with nitrosamine selective detection (GC-TEA). Levels of TSNA in most Moldovan cigarettes were substantially lower than in American brands. Mean levels of NNN in three commercial American brands were 3.32 +/- 0.88 (SD) microg/g as compared to 0.579 +/- 0.548 microg/g, range 0.093-2.09 microg/g (N = 18), in the cigarettes produced in Moldova. For NNK and NAT, mean levels in the American brands were 1.57 +/- 0.178 and 1.99 +/- 0.579 microg/g, respectively, while the corresponding values for Moldovan cigarettes were 0.193 +/- 0.089, range 0.104-0.484 microg/g, and 0.160 +/- 0.114 microg/g, range 0.055-0.481 microg/g. The highest levels of NNN-1.10-2.09 microg/g-were observed in "American type" cigarettes manufactured from high-quality tobacco. The results of this study should be useful in heightening the awareness of the dangers of smoking in Moldova and can be envisioned as the initial step in the control of tobacco-related cancer in this republic.


High intracellular c-di-GMP levels antagonize quorum sensing and virulence gene expression in Burkholderia cenocepacia H111.
The opportunistic human pathogen Burkholderia cenocepacia H111 uses two chemically distinct signal molecules for controlling gene expression in a cell density-dependent manner: N-acyl-homoserine lactones (AHLs) and cis-2-dodecenoic acid (BDSF). Binding of BDSF to its cognate receptor RpfR lowers the intracellular c-di-GMP level, which in turn leads to differential expression of target genes. In this study we analysed the transcriptional profile of B. cenocepacia H111 upon artificially altering the cellular c-di-GMP level. One hundred and eleven genes were shown to be differentially expressed, 96 of which were downregulated at a high c-di-GMP concentration. Our analysis revealed that the BDSF, AHL and c-di-GMP regulons overlap for the regulation of 24 genes and that a high c-di-GMP level suppresses expression of AHL-regulated genes. Phenotypic analyses confirmed changes in the expression of virulence factors, the production of AHL signal molecules and the biosynthesis of different biofilm matrix components upon altered c-di-GMP levels. We also demonstrate that the intracellular c-di-GMP level determines the virulence of B. cenocepacia to Caenorhabditis elegans and Galleria mellonella.


The ribonucleic acids of the mammary gland of the guinea pig.
1. 32-P-labelled polyribosome preparations were made from the mammary glands of lactating and late-pregnant guinea pigs after injection of (32-P)i into the animals. 2. The RNA of polyribosomes, ribosomal subunits and that released from polyribosomes by EDTA were analysed by zone velocity centrifugation and by polyacrylamide-gel electrophoresis. 3. RNA species which have the physical properties expected for the milk protein mRNA were detected. RNA species of a size which could code for the caseins were present in lactating but not in pre-lactating mammary-gland polyribosomes.


Urinary Sugars--A Biomarker of Total Sugars Intake.
Measurement error in self-reported sugars intake may explain the lack of consistency in the epidemiologic evidence on the association between sugars and disease risk. This review describes the development and applications of a biomarker of sugars intake, informs its future use and recommends directions for future research. Recently, 24 h urinary sucrose and fructose were suggested as a predictive biomarker for total sugars intake, based on findings from three highly controlled feeding studies conducted in the United Kingdom. From this work, a calibration equation for the biomarker that provides an unbiased measure of sugars intake was generated that has since been used in two US-based studies with free-living individuals to assess measurement error in dietary self-reports and to develop regression calibration equations that could be used in future diet-disease analyses. Further applications of the biomarker include its use as a surrogate measure of intake in diet-disease association studies. Although this biomarker has great potential and exhibits favorable characteristics, available data come from a few controlled studies with limited sample sizes conducted in the UK. Larger feeding studies conducted in different populations are needed to further explore biomarker characteristics and stability of its biases, compare its performance, and generate a unique, or population-specific biomarker calibration equations to be applied in future studies. A validated sugars biomarker is critical for informed interpretation of sugars-disease association studies.


Insulin-like growth factor-1 induces lipid production in human SEB-1 sebocytes via sterol response element-binding protein-1.
An understanding of the molecular signaling involved in sebaceous gland lipid production is needed to develop therapeutic targets to improve acne. Treatment with methylisobutylxanthine, dexamethasone, and a high dose of insulin (MDI) has been shown to differentiate 3T3-L1 preadipocytes into adipocytes, a differentiation marked by an increase in lipid production. The present study has the following aims: (1) Since high doses of insulin, as found in MDI, will activate the IGF-1 receptor, we sought to determine if IGF-1 is capable of reproducing the lipogenic effect seen with MDI treatment, and (2) to determine if the sterol response element-binding protein-1 (SREBP-1) pathway mediates the increase in lipogenesis. Here we report that MDI increases lipogenesis and that this effect can be attributed wholly to the high-dose insulin in SEB-1 cells. Further, we show that a physiologically relevant dose of IGF-1 or high-dose (1 microM) insulin induces an increase in SREBP-1 mRNA, protein, and total lipid production; while 100 nM insulin induces lipogenesis yet the SREBP protein levels remain unchanged. These data indicate that activation of the IGF-1 receptor increases lipogenesis in SEB-1 cells through both SREBP-dependent and SREBP-independent pathways.


Improved synergic therapeutic effects of chemoradiation therapy with the aid of a co-drug-loaded nano-radiosensitizer under conventional-dose X-ray irradiation.
The goal of this work is to harness the advantages of a targeted hybrid nanostructure, BSA-coated Fe3O4 (F)-Au heterodimer, as a radiosensitizer and co-delivery vehicle of chemotherapeutic drugs for enhanced synergic cancer therapy and protection of healthy tissues. F-Au-BSA-MTX-CUR combines the abilities of enhanced X-ray radiation therapy (F-Au), long blood circulation time (BSA), tumor targeting (MTX), enhanced chemotherapy (MTX and CUR), and protection of normal cells against the harmful effects of radiation (CUR). In this work, we present the radioprotective and radiosensitizing effects of CUR on normal tissues and the tumor site, respectively. After technical evaluation, drug loading, drug release behavior, hemolysis assay, transfection efficacy, and cellular uptake studies with fluorescence microscopy, the biosafety and toxicity of the nanostructure was assessed in vitro and in vivo. Also, to confirm its power to improve synergistic chemoradiation therapy in mice, the antitumor effects of the designed treatment plan were assessed in a 4T1-tumor bearing mouse model. The in vivo antitumor effect evaluation interestingly reveals outstanding therapeutic power of the final formulation (F-Au-BSA-MTX-CUR) and further requirement of CUR as a radioprotective. This result importantly revealed the radioprotection effect of CUR. Co-delivery of the chemotherapeutic drugs MTX and CUR, combined with the radiosensitizing effect of the F-Au heterodimer and the radioprotective effect of CUR, showed promising prospects in cancer therapy.


Can Polyhydroxyalkanoates Be Produced Efficiently From Waste Plant and Animal Oils?
Polyhydroxyalkanoates (PHAs) are a potential replacement for some petrochemical-based plastics. PHAs are polyesters synthesized and stored by various bacteria and archaea in their cytoplasm as water-insoluble inclusions. PHAs are usually produced when the microbes are cultured with nutrient-limiting concentrations of nitrogen, phosphorus, sulfur, or oxygen and excess carbon sources. Such fermentation conditions have been optimized by industry to reduce the cost of PHAs produced commercially. Industrially, these biodegradable polyesters are derived from microbial fermentation processes utilizing various carbon sources. One of the major constraints in scaling-up PHA production is the cost of the carbon source metabolized by the microorganisms. Hence, cheap and renewable carbon substrates are currently being investigated around the globe. Plant and animal oils have been demonstrated to be excellent carbon sources for high yield production of PHAs. Waste streams from oil mills or the used oils, which are even cheaper, are also used. This approach not only reduces the production cost for PHAs, but also makes a significant contribution toward the reduction of environmental pollution caused by the used oil. Advancements in the genetic and metabolic engineering of bacterial strains have enabled a more efficient utilization of various carbon sources, in achieving high PHA yields with specified monomer compositions. This review discusses recent developments in the biosynthesis and classification of various forms of PHAs produced using crude and waste oils from the oil palm and fish industries. The biodegradability of the PHAs produced from these oils will also be discussed.


USP7 promotes decidualization of ESCs by STAT3/PR axis during early pregnancy.
Decidualization is an essential process for embryo implantation during early pregnancy. Defective decidualization is a critical leading cause of early pregnancy loss (EPL). Ubiquitin-specific protease 7 (USP7) is a deubiquitinating enzyme that is involved in uterine function. This study aimed to explore the underlying mechanism by which USP7 regulates decidualization in EPL. We found that USP7 was downregulated in the decidual tissue of EPL patients. Upregulation of USP7 enhanced decidualization in endometrial stromal cells (ESCs), with increased decidualized biomarkers IGFBP1 and PRL and progesterone receptor A/B (PR-A/B) expression. Moreover, we found that signal transducer and activator of transcription 3 (STAT3) is a direct target of USP7 in ESCs. USP7 bound to STAT3 by deubiquitination and increased STAT3 levels in ESCs. Suppression of STAT3 impeded the USP7-promoted cell viability, decidualization, and PR-A/B expression of ESCs. USP7 promoted the decidualization of ESCs through the STAT3/PR signaling pathway during early pregnancy, which provides new insight into the pathological mechanism of EPL and may contribute to the clinical treatment of EPL.


Effects of temperature and conditioning on contact lens wetting angles.
Because wettability is not always examined under standard conditions, we investigated the temperature dependence of saline wettability on unconditioned and conditioned polymethylmethacrylate (PMMA), cellulose acetate butyrate (CAB), and three silicone acrylate lens materials. Sessile drop contact angles were measured in a humidity chamber at 23 degrees C and 34 degrees C using laser-assisted contact angle goniometry. In separate experiments, saline-stored and preconditioned lenses were examined either with or without rinsing. Sessile drop contact angles at 34 degrees C were within 2 degrees to 5 degrees of the room temperature values for both conditioned and unconditioned lenses, demonstrating a negligible temperature dependence. At both temperatures, the conditioned PMMA, CAB, silafocon A, and pasifocon C lenses wet slightly better, by 1 degree to 12 degrees, than unconditioned lenses. However, this increase was only significant with PMMA and silafocon A (P less than 0.05) and reversed when the preconditioned lenses were rinsed repeatedly in saline and reexamined. The results suggest that for these materials: 1) in vitro saline contact angles do not approach those seen on the eye, and this discrepancy can not be explained by temperature or conditioning; and 2) conditioning does not increase material wettability but merely forms a temporary hydrophilic interface that is more wettable than the lens material.


New hexatic liquid phase observed in lyotropic thin films.
An intermediate surface hexatic phase between the liquid and the crystalline phases has been found for the first time in a lyotropic lamellar liquid-crystal system. This phase is highly unusual in that it has long-range sixfold bond-orientational order but liquidlike nearest-neighbor positional correlations, and could represent a significant departure from our current understanding of defect-mediated melting in two dimensions.


The logistics of voucher management: the underreported component in family planning voucher discussions.
The purpose of health care vouchers or coupons is to receive a health service in exchange which is fully or partially subsidized, such as any treatment offered for communicable disease; for immunization; antenatal care-/postnatal care-related maternal health services; a family planning (FP) service; or to get a health commodity like a medicine. Vouchers are targeted for a group of people who can benefit the most such as on the basis of poverty ranking, marginalized or living in rural areas. According to the World Health Organization, voucher schemes in the area of sexual and reproductive health are considered of high value if they are implemented to address the issues of contraceptive commodity or service unavailability or to address the barriers to access such services through contracting out health services, for example, through social franchising (SF). FP vouchers can substantially expand contraceptive access and choice and empower the underserved populations. Literature cites voucher's effectiveness in better targeting, increasing use, and improving program outcomes in FP programs; however, there is little research or explanation of how voucher management is done in practice. The paper attempts to describe various components of voucher management system and its functioning using example of a voucher program in Pakistan. There are challenges such as high upfront cost, targeting the appropriate clients, validation of vouchers, and quality assurance, but these can be managed with better preparation at the planning and design stage. Strong monitoring and evaluation are integral to successful implementation of the voucher program. Also, voucher interventions that are targeted and adopt a pro-poor strategy have been found to improve access to care within poor and marginalized populations. Such programs have the capacity to bridge health inequities in developing nations. Targeted voucher schemes such as those which are designed as pro-poor or pro-rural are known to reduce barriers to access for those living with poverty or for the ones considered as marginalized population. Hence, such interventions have the capacity to fulfill the gaps in health inequities, especially, in low- and/or middle-income countries. Voucher programs should report the voucher logistics and management to build a larger evidence base of best practices. All voucher schemes must be designed, implemented, and evaluated on the basis of set objectives through addressing the local context. But any voucher implementing organization also conducting the in-house voucher management simultaneously may be considered as a weakness in program design, in turn providing rationale for either failure or success of that particular voucher intervention. Therefore, separating implementation and management of a voucher initiative can lead to enhanced transparency, improved accountability, allow for independent validation of services, and facilitate compliance for payments.


Stanniocalcin-1 regulates re-epithelialization in human keratinocytes.
Stanniocalcin-1 (STC1), a glycoprotein hormone, is believed to be involved in various biological processes such as inflammation, oxidative responses and cell migration. Riding on these emerging evidences, we hypothesized that STC1 may participate in the re-epithelialization during wound healing. Re-epithelialization is a critical step that involves keratinocyte lamellipodia (e-lam) formation, followed by cell migration. In this study, staurosporine (STS) treatment induced human keratinocyte (HaCaT) e-lam formation on fibronectin matrix and migration via the activation of focal adhesion kinase (FAK), the surge of intracellular calcium level [Ca²⁺]i and the inactivation of Akt. In accompanied with these migratory features, a time- and dose-dependent increase in STC1 expression was detected. STC1 gene expression was found not the downstream target of FAK-signaling as illustrated by FAK inhibition using PF573228. The reduction of [Ca²⁺]i by BAPTA/AM blocked the STS-mediated keratinocyte migration and STC1 gene expression. Alternatively the increase of [Ca²⁺]i by ionomycin exerted promotional effect on STS-induced STC1 gene expression. The inhibition of Akt by SH6 and GSK3β by lithium chloride (LiCl) could respectively induce and inhibit the STS-mediated e-lam formation, cell migration and STC1 gene expression. The STS-mediated e-lam formation and cell migration were notably hindered or induced respectively by STC1 knockdown or overexpression. This notion was further supported by the scratched wound assay. Collectively the findings provide the first evidence that STC1 promotes re-epithelialization in wound healing.


Multiphysics simulation of ion concentration polarization induced by a surface-patterned nanoporous membrane in single channel devices.
Microfluidic devices utilize ion concentration polarization (ICP) phenomena for a variety of applications, but a comprehensive understanding of the generation of ICP is still necessary. Recently, the emergence of a novel single channel ICP (SC-ICP) device has stimulated further research on the mechanism of ICP generation, so that we developed a 2-D model of an SC-ICP device that integrates a nanoporous membrane on the bottom surface of the channel, allowing bulk flow over the membrane. We solved a set of coupled governing equations with appropriate boundary conditions to explore ICP numerically. As a result, we not only showed that the simulation results held a strong qualitative agreement with experimental results, but also found the distribution of ion concentrations in the SC-ICP device that has never been reported in previous studies. We confirmed again that the electrophoretic mobility (EPM) of counterions in the membrane is the most dominant factor determining the generation and strength of ICP, whereas the charge density of the membrane was dominant to the ICP strength only when a high EPM value was assumed. From the viewpoint of practical applications, an SC-ICP device with a long membrane under low buffer strength showed enhanced performance in the preconcentration of charged molecules. Therefore, we believe that the simulation results could not only provide sharp insight into ICP phenomena but also predict and optimize the performance of SC-ICP devices in various microfluidic applications.


Gross intraoperative evaluation (GIE): a reliable method for the evaluation of surgical margins at partial nephrectomy.
To determine the efficacy of a new method called by us as "gross intra-operative evaluation (GIE)" for the assessment of surgical margin (SM) status. A total of 26 consecutive patients operated with cT1a-b renal tumors at a single center were included in this study. After the excision, the tumors were uniformly divided into two halves in the longitudinal axis ex vivo. In this way, margins were exposed for GIE for the evaluation of the safety of SMs. Findings of GIE were compared with the permanent section analysis in terms of SM status. Mean patient age, tumor size and margin thickness was 59 (38-79), 3.1 (1.5-6) cm and 3.7 (0.1-12) mm, respectively. In all patients, GIE showed intact margins and none of the patients had positive SM in the final pathological examination. There was no evidence of local recurrence or distant metastasis with a mean follow-up of 25 (4-104) months. All patients are alive. GIE of resected specimen without FS analysis is a safe and effective method for the evaluation of SMs in partial nephrectomy patients.


Quantitative and qualitative signals determine T-cell cycle entry and progression.
Cell growth and proliferation as well as cell cycle arrest and apoptosis all play integral roles in the cellular immune response. The signals that lead to cytokine production by antigen- or mitogen-stimulated T cells have been studied in detail. However, it is not fully understood how these signals promote cell cycle entry and progression to DNA synthesis in T lymphocytes, especially in primary cells. We used a model distinguishing between competence and progression phases to examine quantitative and qualitative differences in signal transduction that resulted in cell cycle entry and G1 phase arrest or led to DNA synthesis in human T cells. Resting peripheral blood T cells were rendered competent by stimulation with submitogenic concentrations of phytohemagglutinin (PHA) or they were stimulated to proliferate using mitogenic concentrations of PHA. The competent state (that is, the capacity to proliferate in response to exogenous IL-2) was characterized by calcium mobilization, a protein kinase C-dependent internalization of CD3, increased mitogen-activated protein kinase (MAPK) activity, transient translocation of AP-1 transcription factors to the nucleus, expression of immediate early genes, activation of G1-phase cyclin-dependent kinases, and increased CD25 (IL-2Ralpha) expression. However, all of these events were of lesser magnitude in T cells rendered competent than in T cells stimulated to proliferate. Furthermore, the mitogenic stimulus induced a different pattern of MAPK activation and sustained translocation of AP-1 to the nucleus with concomitant IL-2 production. The data indicate that quantitative and qualitative differences in early signaling events distinguish the acquisition of the competent state or the induction of cytokine production with a commitment to T-cell proliferation.


Mitochondrial dysfunction and endoplasmic reticulum stress in calf hepatocytes are associated with fatty acid-induced ORAI calcium release-activated calcium modulator 1 signaling.
The store-operated Ca2+ entry (SOCE) moiety ORAI calcium release-activated calcium modulator 1 (ORAI1) located in the endoplasmic reticulum (ER) participates in key cellular functions such as protein folding, transport, and secretion, and lipid metabolism. We used an in vitro approach to test whether exogenous fatty acids alter ORAI1 signaling and to explore potential consequences on mitochondrial dysfunction and ER stress. First, hepatocytes isolated from 4 healthy female calves (1 d old, 40-50 kg) were challenged with a 1.2 mM mixture of oleic, linoleic, palmitic, stearic, and palmitoleic acids for 0.5, 1, 3, 6, 9, and 12 h to measure oxidative stress [intracellular reduced glutathione (GSH), superoxide dismutase (SOD), malondialdehyde (MDA), and hydrogen peroxide] and ER stress (protein abundance of PERK, IRE, ATF6, and GRP78). Concentrations of GSH and SOD decreased at 0.5 h, and MDA and hydrogen peroxide increased at 1 h; ER stress proteins increased at 6 h. To determine whether ER stress was caused by oxidative stress, primary calf hepatocytes were treated with the same 1.2 mM fatty acid mix or the reactive oxygen species (ROS) inhibitor N-acetylcysteine (NAC) for 6 h. We found that NAC prevented an increase in ER stress protein abundance. Next, the role of ORAI1 on ER stress was measured by transfecting hepatocytes with small interfering (si)ORAI1 or the ORAI1 inhibitor BTP2, followed by a challenge with 1.2 mM fatty acids for 3 h. Without inhibiting ORAI1, exogenous fatty acids upregulated ORAI1 mRNA and protein abundance, oxidative stress, ER stress proteins, and protein abundance of marker indicators of an opened mitochondrial permeability transition pore (mPTP). Inhibition with BPT2 or silencing via siORAI1 abrogated oxidative stress, including increased GSH concentration and SOD activity, decreased MDA, hydrogen peroxide, and ROS concentration; ER stress protein abundance was downregulated, and mitochondrial function was restored. Last, changes in markers of mPTP opening were evaluated by culturing hepatocytes for 6 h with the sarcoendoplasmic Ca2+ ATPase inhibitor thapsigargin or the calcium ionophore ionomycin. We detected an increase in VDAC1, CLPP, and CypD protein abundance, all of which indicated opening of the mPTP. Overall, data from these in vitro studies suggest that ORAI1 mediates ER stress induced by high concentrations of fatty acids, in part through alleviating mitochondrial dysfunction caused by oxidative stress.


[Improvement of ultrasound study in the diagnosis of pleurisies of various etiologies].
The study was undertaken to improve ultrasonography using needle biopsy under ultrasound control in the diagnosis of pleurisies of various etiologies. The patients of the Department of Thoracic Surgery, who had been divided by the nosological principle (nonspecific pleurisy, tuberculosis effusions, and tumorous and posttraumatic pleurisies), were followed up. The study and statistical processing yielded major ultrasound criteria for pleural effusions and a final diagnosis was made on the basis of needle biopsy under ultrasound control with subsequent morphological verification.