Medium-dependent control of the bacterial growth rate.
By combining results from previous studies of nutritional up-shifts we here re-investigate how bacteria adapt to different nutritional environments by adjusting their macromolecular composition for optimal growth. We demonstrate that, in contrast to a commonly held view the macromolecular composition of bacteria does not depend on the growth rate as an independent variable, but on three factors: (i) the genetic background (i.e. the strain used), (ii) the physiological history of the bacteria used for inoculation of a given growth medium, and (iii) the kind of nutrients in the growth medium. These factors determine the ribosome concentration and the average rate of protein synthesis per ribosome, and thus the growth rate. Immediately after a nutritional up-shift, the average number of ribosomes in the bacterial population increases exponentially with time at a rate which eventually is attained as the final post-shift growth rate of all cell components. After a nutritional up-shift from one minimal medium to another minimal medium of higher nutritional quality, ribosome and RNA polymerase syntheses are co-regulated and immediately increase by the same factor equal to the increase in the final growth rate. However, after an up-shift from a minimal medium to a medium containing all 20 amino acids, RNA polymerase and ribosome syntheses are no longer coregulated; a smaller rate of synthesis of RNA polymerase is compensated by a gradual increase in the fraction of free RNA polymerase, possibly due to a gradual saturation of mRNA promoters. We have also analyzed data from a recent publication, in which it was concluded that the macromolecular composition in terms of RNA/protein and RNA/DNA ratios is solely determined by the effector molecule ppGpp. Our analysis indicates that this is true only in special cases and that, in general, medium adaptation also depends on factors other than ppGpp.


Sutures, incisions, and anastomoses.
The properties of different types of suture material are reviewed, with particular reference to the complications which may result from their use. The aetiology, prevention, diagnosis, and treatment of wound dehiscence are discussed. The factors which are believed to be important in performing successful bowel anastomosis are considered.


Stress and burnout in community mental health nursing: a review of the literature.
There is a growing body of evidence that suggests that many community mental health nurses (CMHNs) experience considerable stress and burnout. This review aimed to bring together the research evidence in this area for CMHNs working within the UK. Seventeen papers were identified in the literature, seven of which looked at stress and burnout for all members of community mental health teams (CMHTs) and the remaining 10 papers focused on CMHNs. The evidence indicates that those health professionals working as part of community teams are experiencing increasing levels of stress and burnout as a result of increasing workloads, increasing administration and lack of resources. For CMHNs specific stressors were identified. These included increases in workload and administration, time management, inappropriate referrals, safety issues, role conflict, role ambiguity, lack of supervision, not having enough time for personal study and NHS reforms, general working conditions and lack of funding and resources. Areas for future research are described and the current study of Welsh CMHNs is announced. This review has been completed against a background of further significant changes in the health service. In the mental health field, specific new initiatives will have a significant impact on the practice of community mental health nursing. A new National Framework for Mental Health, along with a review of the Mental Health Act (1983), will undoubtedly help to shape the future practice of mental health nursing.


Gene expression patterns of Salpa thompsoni reveal remarkable differences in metabolism and reproduction near the Antarctic Polar Front.
Salpa thompsoni is an important grazer in the Southern Ocean and most abundant in the Antarctic Polar Front (APF) region. During recent decades, their distribution expanded southwards. However, it is unclear whether salps can maintain their populations in the high Antarctic regions throughout the year owing to a poor understanding of their physiological responses to changing environmental conditions. We examined gene expression signatures of salps collected in two geographically close regions south of the APF that differed in water mass composition and productivity. The observed differences in the expression of genes related to reproductive, cellular and metabolic processes reflect variations in water temperature and food supply between the two regions studied here. Our study contributes to a better understanding of the physiological responses of S. thompsoni to changing environmental conditions, and how the species may adapt to a changing environment through potential geographical population shifts under future climate change scenarios.


Job satisfaction of psychiatric hospital employees: a new measure of an old concern.
The authors report on the factor structure of an employee satisfaction questionnaire designed for use with psychiatric hospital employees. The actions and attitudes of management were, by far, the single most prominent factor. This factor captures the extent to which management respects workers, operates with honesty and integrity, promotes efficiency, and has open lines of communication with employees. By surveying employees about the 15 items of this factor, psychiatric hospital managers can obtain a sense of their employees' overall satisfaction. Hospitals must hire and train managers who can convey the qualities represented in this satisfaction factor. It is also important for hospital managers to recognize that employee satisfaction is strongly related to how employee complaints are handled, how employees perceive the quality of patient care, and the extent to which employees believe the hospital serves the greater community.


The pain threshold of high-threshold mechanosensitive receptors subsequent to maximal eccentric exercise is a potential marker in the prediction of DOMS associated impairment.
Delayed-onset muscle soreness (DOMS) refers to dull pain and discomfort in people after participating in exercise, sport or recreational physical activities. The aim of this study was to detect underlying mechanical thresholds in an experimental model of DOMS. Randomised study to detect mechanical pain thresholds in a randomised order following experimentally induced DOMS of the non-dominant arm in healthy participants. Main outcome was the detection of the pressure pain threshold (PPT), secondary thresholds included mechanical detection (MDT) and pain thresholds (MPT), pain intensity, pain perceptions and the maximum isometric voluntary force (MIVF). Twenty volunteers (9 female and 11 male, age 25.2 ± 3.2 years, weight 70.5 ± 10.8 kg, height 177.4 ± 9.4 cm) participated in the study. DOMS reduced the PPT (at baseline 5.9 ± 0.4 kg/cm2) by a maximum of 1.5 ± 1.4 kg/cm2 (-24%) at 48 hours (p < 0.001). This correlated with the decrease in MIVF (r = -0.48, p = 0.033). Whereas subjective pain was an indicator of the early 48 hours, the PPT was still present after 72 hours (r = 0.48, p = 0.036). Other mechanical thresholds altered significantly due to DOMS, but did show no clinically or physiologically remarkable changes. Functional impairment following DOMS seems related to the increased excitability of high-threshold mechanosensitive nociceptors. The PPT was the most valid mechanical threshold to quantify the extent of dysfunction. Thus PPT rather than pain intensity should be considered a possible marker indicating the athletes' potential risk of injury.


Predicting Adverse Drug Reactions on Distributed Health Data using Federated Learning.
Using electronic health data to predict adverse drug reaction (ADR) incurs practical challenges, such as lack of adequate data from any single site for rare ADR detection, resource constraints on integrating data from multiple sources, and privacy concerns with creating a centralized database from person-specific, sensitive data. We introduce a federated learning framework that can learn a global ADR prediction model from distributed health data held locally at different sites. We propose two novel methods of local model aggregation to improve the predictive capability of the global model. Through comprehensive experimental evaluation using real-world health data from 1 million patients, we demonstrate the effectiveness of our proposed approach in achieving comparable performance to centralized learning and outperforming localized learning models for two types of ADRs. We also demonstrate that, for varying data distributions, our aggregation methods outperform state-of-the-art techniques, in terms of precision, recall, and accuracy.


[Diabetes therapy in pregnant women].
Diabetes type 2 has increased in adult women due to higher frequency of obesity and sedentary lifestyle, and thus cardiovascular diseases have increased. Hyperglycemia control and, collaterally, high blood pressure and dyslipidemia correction are the basis of its therapy. This paper evaluates oral antidiabetic drugs effectiveness and preference. Gestational diabetes is a cause of morbidity and mortality in fetuses and newborns, it has to be timely diagnosed, and needs a strict control, same to diabetes type 1. Gyneco-obstetrical doctor must be alert to diabetes signs and closely work in its therapy.


Effect of calcium deficiency on acyl-CoA synthetase activity in microsomes from various rat tissues.
The specific activity of long-chain acyl-CoA synthetase in microsomes from various tissues of control and calcium-deficient rats was determined. It was found that the saturated acids, palmitic and stearic, were preferential substrates compared to the non-saturated linoleic, alpha-linolenic and eicosa-8,11,14-trienoic acids. All of them showed similar Vm values with different affinity constants. After 60-day treatment on a calcium-deficient diet (0.5 g Ca/Kg diet), a significant increase in the acyl-CoA synthetase activity was observed for all the tested fatty acids in liver and kidney microsomes. These changes were evoked without any modification in the substrate selectivities shown for the control microsomes, and they were well-correlated with calcium level in both tissues. Under the calcium deficient state an increase in Vm values was observed for palmitic and eicosatrienoic acids with no changes in the corresponding Km, suggesting an increment in the number of active enzyme molecules within the microsomal membrane.


Assessment of resident surgical skills: is testing feasible?
We have previously shown that in a single residency program objective structured assessment of technical skills (OSATS) is a reliable and valid method of assessing surgical competency. Our goal was to establish feasibility of this evaluation instrument when administered at multiple residency programs throughout the US, and assess the impact of a laboratory-based surgical curriculum on results. An OSATS exam was administered to 116 residents from 5 residency programs. One of the residency programs had participated in a comprehensive surgical curriculum over a 4-year period of time. The exam consisted of 3 open and 3 laparoscopic tasks. Residents were graded by both a blinded and unblinded examiner with task-specific checklist, global rating scale, pass/fail assessment, and tasks were timed. All tasks were performed on life-like models. Examinations were successfully completed at all sites. Each exam required 90 minutes of resident time. Reliability indices calculated with Cronbach's alpha were .97 for overall global rating and .95 for checklists. Interrater reliability between blinded and unblinded examiners ranged from .71 to .97 for individual tasks and was .95 overall. Assessment of construct validity (the ability to distinguish among residency levels) found significant differences among the residents for both blinded and unblinded examiners for all evaluation outcomes except time. For the test overall, the global rating scale showed significant differences among all 4 residency levels. The checklist showed significant differences at three levels (PGY3-4 >PGY2 >PGY1). Approximate cost for replaceable items was 40 US dollars to 150 per resident depending on which tasks were chosen. Comparison of scores between residents who received a laboratory-based curriculum and those who did not revealed significantly higher scores and shorter time to complete tasks for the group who received additional training. Large-scale testing has confirmed that OSATS is an objective, reliable, and valid method to assess surgical skills, and can easily be administered in most residency programs. A laboratory-based surgical curriculum improved test results and reduced time to complete tasks.


Hyperphosphorylation of pRb: a mechanism for RB tumour suppressor pathway inactivation in bladder cancer.
Loss of heterozygosity, mutations or deletions of the RB1 gene usually result in loss of pRb expression, which has been regarded as an indicator of loss of pRb function in human tumours. It has previously been shown that in addition to loss of pRb expression, aberrantly high (pRb2+) pRb expression also indicates loss of pRb function in bladder tumours compared with moderate (normal, pRb1+) pRb expression. The aim of this study was to elucidate the mechanism by which pRb is functionally inactivated in bladder tumours expressing aberrantly high levels of pRb. Constitutive phosphorylation was therefore investigated as a mechanism of pRb inactivation in bladder tumours. Of 28 bladder tumours examined, western blotting demonstrated pRb hyperphosphorylation in 5/7 (71%) pRb2+ bladder tumours compared with only 4/11 (36%) pRb1+ tumours (p = 0.002). All cases with undetectable pRb showed moderate to high p16 expression and none showed cyclin D1 expression by immunohistochemistry. All pRb1+ tumours with underphosphorylated pRb showed p16 but not cyclin D1 expression. All pRb2+ tumours with hyperphosphorylated pRb showed loss of p16 expression and/or cyclin D1 overexpression. Thus, elevated pRb expression was associated with pRb hyperphosphorylation, which, in turn, was associated with loss of p16 expression and/or increased cyclin D1 expression. In order to analyse this association in vitro, T24 cells, which express high levels of pRb, were transfected with p16 cDNA. Transfection with p16 cDNA resulted in a marked decrease in pRb phosphorylation, decreased cell proliferation, and a change in expression of pRb from high to moderate phenotype as assessed by immunohistochemistry. This paper gives the biological basis for constitutive alteration of pRb function in human tumours in the presence of an intact, expressed pRb protein; the mechanism of pRb inactivation is through hyperphosphorylation, which results from loss of p16 expression and/or cyclin D1 overexpression. Immunohistochemical expression of pRb appears to be a reliable indicator of pRb function.


Trends in unilateral breast reconstruction and management of the contralateral breast: the Emory experience.
Recent trends in breast reconstruction have transitioned toward the skin-sparing type of mastectomy and immediate reconstruction using autologous tissue. This study was designed to document trends in the management of patients with unilateral breast cancer and to determine how they influence management of the contralateral breast. All patients who underwent unilateral breast reconstruction at Emory University Hospitals from January of 1975 to December of 1999 were reviewed. The cohort was stratified by timing of reconstruction (immediate versus delayed), method of reconstruction, and mastectomy type (skin-sparing versus non-skin-sparing). The methods of reconstruction included implant, latissimus dorsi flap, and transverse rectus abdominis musculocutaneous (TRAM) flap. Contralateral procedures to achieve symmetry included augmentation, mastopexy, augmentation/mastopexy, and reduction. A total of 1394 patients were evaluated, including 689 delayed and 705 immediate reconstructions. Sixty-seven percent of delayed-reconstruction patients (462 of 689) had a symmetry procedure performed on the opposite breast, compared with 22 percent for the immediate-reconstruction patients (155 of 705) (p </= 0.001). The percentage of times a contralateral procedure was performed was highest for implant reconstructions (89 percent delayed and 57 percent immediate) and lowest for TRAM flap reconstructions (59 percent delayed and 18 percent immediate). Augmentation mammaplasty was the most common symmetry procedure for implant reconstruction (41 percent), whereas reduction was the most common procedure for autologous tissue reconstruction (57 percent). Immediate unilateral breast reconstructions were stratified into non-skin-sparing mastectomy (n = 205) and skin-sparing mastectomy (n = 500). Thirty-four percent of patients with a non-skin-sparing mastectomy defect (70 of 205) underwent a contralateral breast procedure, compared with 17 percent of patients with a skin-sparing mastectomy defect (85 of 500) (p = 0.001). The percentage of times a contralateral procedure was performed in immediate reconstruction, stratified by mastectomy and reconstruction type, was only significant for TRAM flap reconstructions (25 versus 11 percent). Trends in the management of unilateral breast cancer from delayed to immediate reconstruction and from implants to autologous tissue have reduced the incidence of contralateral symmetry procedures. Reduction mammaplasty is the most common symmetry procedure used for autologous tissue reconstruction, with augmentation predominating when implants are used. The type of mastectomy also effects the management of the opposite breast, with skin-sparing mastectomy further reducing the incidence of contralateral procedures in immediate TRAM flap reconstruction, compared with non-skin-sparing mastectomy.


Reversible and high-contrast thermal conductivity switching in a flexible covalent organic framework possessing negative Poisson's ratio.
The ability to dynamically and reversibly control thermal transport in solid-state systems can redefine and propel a plethora of technologies including thermal switches, diodes, and rectifiers. Current material systems, however, do not possess the swift and large changes in thermal conductivity required for such practical applications. For instance, stimuli responsive materials, that can reversibly switch between a high thermal conductivity state and a low thermal conductivity state, are mostly limited to thermal switching ratios in the range of 1.5 to 4. Here, we demonstrate reversible thermal conductivity switching with an unprecedented 18× change in thermal transport in a highly flexible covalent organic framework with revolving imine bonds. The pedal motion of the imine bonds is capable of reversible transformations of the framework from an expanded (low thermal conductivity) to a contracted (high thermal conductivity) phase, which can be triggered through external stimuli such as exposure to guest adsorption and desorption or mechanical strain. We also show that the dynamic imine linkages endow the material with a negative Poisson's ratio, thus marking a regime of materials design that combines low densities with exceptional thermal and mechanical properties.


Biological Activities of Stilbenoids.
Stilbenoids are a group of naturally occurring phenolic compounds found in various plant species. They share a common backbone structure known as stilbene, but differ in the nature and position of substituents. Stilbenoids are classified as phytoalexins, which are antimicrobial compounds produced de novo in plants to protect against fungal infection and toxins. In this review, the biological effects of stilbenoids such as resveratrol, pterostilbene, gnetol and piceatannol are discussed. Stilbenoids exert various biological activities ranging from cardioprotection, neuroprotection, anti-diabetic properties, depigmentation, anti-inflammation, cancer prevention and treatment. The results presented cover a myriad of models, from cell culture to animal studies as well as clinical human trials. Although positive results were obtained in most cell culture and animal studies, further human studies are needed to substantiate beneficial effects of stilbenoids. Resveratrol remains the most widely studied stilbenoid. However, there is limited information regarding the potential of less common stilbenoids. Therefore, further research is warranted to evaluate the salutary effects of various stilbenoids.


A comparison of Australian families' expenditure on active and screen-based recreation using the ABS Household Expenditure Survey 2003/04.
This study aimed to investigate how much households with dependent children spend on active recreation (physical activity) compared with screen-based (sedentary) recreation, according to their household socioeconomic and demographic characteristics. The study analysed data from the 2003-04 Australian Bureau of Statistics Household Expenditure Survey, which collected information on household expenditure from a representational cross-section of private dwellings across Australia. In 2003-04, Australian households with dependent children spent an average of 1.5% and 3.3% of their weekly disposable income on active and screen recreation respectively, and 24.9% of their total active and screen recreation expenditure on active recreation. There was significant variation across household characteristics, with higher income and socioeconomic status households, and families with more than one dependent child more likely to spend a larger portion of their recreation budget on active recreation instead of screen recreation. Overall, Australian families spend more money on screen recreation items than they do on active recreation, although there are strong economic and cultural gradients in their patterns of expenditure on both active and screen recreation. This suggests that while the costs of active recreation may be a barrier to participation for some families, there are also social and cultural values influencing recreational choices. For the first time, specific information on Australian families' expenditure on active and screen recreation is available. These results contribute to identifying cultural and economic barriers influencing families' health-related behaviours and their participation in organised physical activity.


Crossed-beam experiment for the scattering of low- and intermediate-energy electrons from BF3: A comparative study with XF3 (X = C, N, and CH) molecules.
Absolute differential cross sections (DCSs) for electron interaction with BF3 molecules have been measured in the impact energy range of 1.5-200 eV and recorded over a scattering angle range of 15°-150°. These angular distributions have been normalized by reference to the elastic DCSs of the He atom and integrated by employing a modified phase shift analysis procedure to generate integral cross sections (ICSs) and momentum transfer cross sections (MTCSs). The calculations of DCSs and ICSs have been carried out using an independent atom model under the screening corrected additivity rule (IAM-SCAR). The present elastic DCSs have been found to agree well with the results of IAM-SCAR calculation above 20 eV, and also with a recent Schwinger multichannel calculation below 30 eV. Furthermore, in the comparison with the XF3 (X = B, C, N, and CH) molecules, the elastic DCSs reveal a similar angular distribution which are approximately equal in magnitude from 30 to 200 eV. This feature suggests that the elastic scattering is dominated virtually by the 3-outer fluorine atoms surrounding the XF3 molecules. The vibrational DCSs have also been obtained in the energy range of 1.5-15 eV and vibrational analysis based on the angular correlation theory has been carried out to explain the nature of the shape resonances. Limited experiments on vibrational inelastic scattering confirmed the existence of a shape resonance with a peak at 3.8 eV, which is also observed in the vibrational ICS. Finally, the estimated elastic ICSs, MTCSs, as well as total cross sections are compared with the previous cross section data available.


Radioprotective effect of ursolic acid in radiation-induced impairment of neurogenesis, learning and memory in adolescent BALB/c mouse.
The effect of acute irradiation with 5Gy or fractionated exposure with 0.5Gy continuously for 10days (a total dose of 5Gy) was evaluated in an immature BALB/c mouse model. Radioprotective effect of ursolic acid (at 25mg/kg/daily administered 1h after acute or each of fractionated irradiations, and continuously for 30days) was also investigated. We found that both acute and fractionated irradiation at a total dose of 5Gy did not induce any mortality within 30days after exposure to postnatal day 26 (P26) BALB/c mice, but reduced animal weigh gain in the first few weeks. At 90days after irradiation, the weight of animals with acute irradiation was still significantly lower than the control group; no significant difference though was observed for those fractionatedly exposed mice compared to the control group. Behavioral tests indicated that acute irradiation at 5Gy induced deficits in learning and memory in the contextual fear conditioning test. The memory for novel object recognition was also impaired. Similar changes were not observed in mice with fractionated irradiation. Immunohistochemical study demonstrated clearly that acute and fractionated irradiations induced impairment of neurogenesis in the subgranular zone (SGZ) of the dentate gyrus although fractionated exposure induced much lesser loss of newly generated neurons. Ursolic acid administered at 25mg/kg/daily for 30days after irradiation greatly improved acute irradiation-induced deficits in contextual learning and memory and in novel object recognition memory although it exacerbated radiation-induced reduction of neurogenesis in SGZ.


Adding insult to injury: neural sensitivity to social exclusion is associated with internalizing symptoms in chronically peer-victimized girls.
Despite evidence documenting activation of the social pain network in response to social rejection and its link to temporary distress, far less is known regarding its role in pervasive emotional difficulties. Moreover, research has not considered the intersection between neural activation to experimentally induced social exclusion and naturally occurring social adversity. This study examined an integrated social pain model of internalizing symptoms, which posits that (i) neural sensitivity in the social pain network is associated with internalizing symptoms, (ii) this linkage is more robust in youth with than without a history of social adversity, and (iii) heightened avoidance motivation serves as one pathway linking neural sensitivity and internalizing symptoms. During a functional magnetic resonance imaging scan, 47 adolescent girls (M age = 15.46 years, SD = .35) with well-characterized histories of peer victimization were exposed to social exclusion. Whole-brain analyses revealed that activation to exclusion in the social pain network was associated with internalizing symptoms. As anticipated, this linkage was stronger in chronically victimized than non-victimized girls and was partially accounted for by avoidance motivation. This research indicates the importance of integrating neural, social and psychological systems of development in efforts to elucidate risk for internalizing symptoms among adolescent girls.


Neisseria gonorrhoeae survive intraleukocytic oxygen-independent antimicrobial capacities of anaerobic and aerobic granulocytes in the presence of pyocin lethal for extracellular gonococci.
The resistance of a piliated, transparent variant of Neisseria gonorrhoeae FA19 to intraleukocytic killing by human polymorphonuclear neutrophils (PMN) was examined. In both aerobic and anaerobic PMN monolayers, approximately 2% of the intracellular gonococci survived for as long as 165 min. Anaerobic PMN were as effective as aerobic PMN in the intracellular killing of gonococci. Hence, O2-independent antimicrobial systems of PMN performed a significant role in the intraleukocytic killing of gonococci were intracellular was supported by the elimination of extracellular bacteria by the addition of pyocin 103 and confirmed by the fluorescent antibody staining of intact gonococci after the PMN were permeabilized to antibody with a Formalin-acetone treatment of PMN monolayers. Our data indicate that while the majority of ingested gonococci are killed by O2-independent antimicrobial systems, a small number (about 2%), survive even when care is taken to eliminate extracellular bacteria.


The acute effects of grape polyphenols supplementation on endothelial function in adults: meta-analyses of controlled trials.
The acute effects of grape polyphenols on endothelial function in adults are inconsistent. Here, we performed meta-analyses to determine these acute effects as measured by flow-mediated dilation (FMD). Trials were searched in PubMed, Embase and the Cochrane Library database. Summary estimates of weighted mean differences (WMDs) and 95% CIs were obtained by using random-effects models. Meta-regression and subgroup analyses were performed to identify the source of heterogeneity. The protocol details of our meta-analysis have been submitted to the PROSPERO register and our registration number is CRD42013004157. Nine studies were included in the present meta-analyses. The results showed that the FMD level was significantly increased in the initial 120 min after intake of grape polyphenols as compared with controls. Meta-regression and subgroup analyses were performed and showed that a health status was the main effect modifier of the significant heterogeneity. Subgroups indicated that intake of grape polyphenols could significantly increase FMD in healthy subjects, and the increased FMD appeared to be more obviously in subjects with high cardiovascular risk factors. Moreover, the peak effect of grape polyphenols on FMD in healthy subjects was found 30 min after ingestion, which was different from the effect in subjects with high cardiovascular risk factors, in whom the peak effect was found 60 min after ingestion. Endothelial function can be significantly improved in healthy adults in the initial 2 h after intake of grape polyphenols. The acute effect of grape polyphenols on endothelial function may be more significant but the peak effect is delayed in subjects with a smoking history or coronary heart disease as compared with the healthy subjects.


Matrix metalloproteinase 9 is a mediator of epidermal growth factor-dependent e-cadherin loss in ovarian carcinoma cells.
Epidermal growth factor (EGF) receptor (EGFR) is frequently elevated in epithelial ovarian cancer, and E-cadherin expression is often reduced in advanced disease. In this study, we investigated a mechanism by which EGFR activation promotes disruption of adherens junctions through induction of matrix metalloproteinase 9 (MMP-9). We show that EGFR activation down-modulates E-cadherin, and broad spectrum MMP inhibition ameliorates EGF-stimulated junctional disruption and loss of E-cadherin protein. MMP-9 involvement in EGF-dependent down-regulation of E-cadherin was determined by siRNA specifically directed against MMP-9. Furthermore, treatment with recombinant MMP-9 or transient expression of MMP-9 is sufficient to reduce E-cadherin levels in differentiated ovarian tumor cells. Stable overexpression of MMP-9 led to a loss of E-cadherin and junctional integrity, and promoted a migratory and invasive phenotype. Thus, elevated MMP-9 protein expression is sufficient for junctional disruption and loss of E-cadherin in these cells. The associations between EGFR activation, MMP-9 expression, and E-cadherin were investigated in human ovarian tumors and paired peritoneal metastases wherein immunohistochemical staining for activated (phospho) EGFR and MMP-9 colocalized with regions of reduced E-cadherin. These data suggest that regulation of MMP-9 by EGFR may represent a novel mechanism for down-modulation of E-cadherin in ovarian cancer.


Using sequence clustering to identify clinically relevant subphenotypes in patients with COVID-19 admitted to the intensive care unit.
The novel coronavirus disease 2019 (COVID-19) has heterogenous clinical courses, indicating that there might be distinct subphenotypes in critically ill patients. Although prior research has identified these subphenotypes, the temporal pattern of multiple clinical features has not been considered in cluster models. We aimed to identify temporal subphenotypes in critically ill patients with COVID-19 using a novel sequence cluster analysis and associate them with clinically relevant outcomes. We analyzed 1036 confirmed critically ill patients with laboratory-confirmed SARS-COV-2 infection admitted to the Mount Sinai Health System in New York city. The agglomerative hierarchical clustering method was used with Levenshtein distance and Ward's minimum variance linkage. We identified four subphenotypes. Subphenotype I (N = 233 [22.5%]) included patients with rapid respirations and a rapid heartbeat but less need for invasive interventions within the first 24 hours, along with a relatively good prognosis. Subphenotype II (N = 418 [40.3%]) represented patients with the least degree of ailments, relatively low mortality, and the highest probability of discharge from the hospital. Subphenotype III (N = 259 [25.0%]) represented patients who experienced clinical deterioration during the first 24 hours of intensive care unit admission, leading to poor outcomes. Subphenotype IV (N = 126 [12.2%]) represented an acute respiratory distress syndrome trajectory with an almost universal need for mechanical ventilation. We utilized the sequence cluster analysis to identify clinical subphenotypes in critically ill COVID-19 patients who had distinct temporal patterns and different clinical outcomes. This study points toward the utility of including temporal information in subphenotyping approaches.


The in vivo effect of bryostatin-1 on paclitaxel-induced tumor growth, mitotic entry, and blood flow.
Pretreatment of tumor cells with the protein kinase C (PKC) inhibitor bryostatin-1 enhances the cytotoxicity of most chemotherapeutic agents. However, in the case of paclitaxel, this effect has been shown in vitro to be best achieved when bryostatin-1 follows (rather than precedes) paclitaxel treatment. With combination trials of bryostatin-1 and paclitaxel planned for clinical trials and with only in vitro data available regarding drug sequence, we elected to undertake an in vivo study evaluating the effect of sequential bryostatin-1 and paclitaxel in a tumor-bearing mouse model and to correlate this effect to cell cycle events, tumor metabolism, and tumor blood flow. At the maximum tolerated i.p. dose, bryostatin-1 at 80 microg/kg resulted in a small but significant increase in tumor doubling time (4.2 +/- 0.3 days) compared with control tumors (3.0 +/- 0.3 days; P < 0.01). Mice treated with i.v. paclitaxel, administered at a dose of 12 mg/kg every 12 h for three doses, weekly for 3 weeks, had a tumor doubling time of 23.4 +/- 1.7 days. Mice pretreated with i.p. bryostatin-1 (80 microg/kg) followed 12 h later by i.v. paclitaxel (12 mg/kg every 12h for three doses) weekly for 3 weeks had a tumor doubling time of 9.7 +/- 1.1 days. This was significantly less (P < .001) than paclitaxel alone, which indicated an inhibitory effect by bryostatin-1 on paclitaxel therapy. In comparison, tumor-bearing mice that were treated with the same dose but with the sequence of paclitaxel followed by bryostatin-1 had a tumor doubling time of 29.6 +/- 0.6 days. This was significantly greater than the tumor doubling times for any condition tested (P < 0.01), demonstrating the sequence dependence of this combination. The efficacy of paclitaxel is dependent on mitotic entry, a step that requires activation of p34cdc2 kinase activity. Treatment with paclitaxel in vivo increased p34 cdc2 kinase activity in the mouse mammary tumors, whereas administration of bryostatin-1 before paclitaxel prevented the p34cdc2 kinase activation by paclitaxel. This was further evaluated in vitro by flow cytometry in MKN-74 human gastric cancer cells. As determined by MPM-2 labeling, which identifies cells in mitosis, pretreatment with bryostatin-1 prevented paclitaxel-treated cells from entering mitosis. Bryostatin-1 has been reported to induce changes in muscle metabolism and to decrease muscle blood flow. These events could impact on the interaction of bryostatin-1 with paclitaxel. Using proton-decoupled phosphorus nuclear magnetic resonance (31P-NMR) spectroscopy in vivo, bryostatin-1 at 80 micro1g/kg induced a decrease in both intratumoral pH and high-energy phosphates. In vivo perfusion studies, using dynamic enhanced NMR imaging with gadolinium diethylenetriamine pentaacetic acid, also demonstrated decreased tumor blood flow. These studies suggest that the inhibition of tumor response to paclitaxel by bryostatin-1 is multifactorial and includes such diverse factors as inhibition of cell entry into mitosis, a decrease in pH and energy metabolism, and a decrease in tumor blood flow. These results indicate that, as this combination enters Phase I clinical trials, the sequence of paclitaxel followed by bryostatin-1 will be critical in the clinical trial design.


Efficacy of decontaminants and disinfectants against Clostridium difficile.
Clostridium difficile is a common nosocomial pathogen transmitted mainly via its spores. These spores can remain viable on contaminated surfaces for several months and are resistant to most commonly used cleaning agents. Thus, effective decontamination of the environment is essential in preventing the transmission of C. difficile in health-care establishments. However, this emphasis on decontamination must also be extended to laboratories due to risk of exposure of staff to potentially virulent strains. Though few cases of laboratory-acquired infection have been reported, the threat of infection by C. difficile in the laboratory is real. Our aim was to test the efficacy of four disinfectants, Actichlor, MicroSol 3+, TriGene Advance and Virkon, and one laboratory decontaminant, Decon 90, against vegetative cells and spores of C. difficile. Five strains were selected for the study: the three most commonly encountered epidemic strains in Scotland, PCR ribotypes 106, 001 and 027, and control strains 630 and VPI 10463. MICs were determined by agar dilution and broth microdilution. All the agents tested inhibited the growth of vegetative cells of the selected strains at concentrations below the recommended working concentrations. Additionally, their effect on spores was determined by exposing the spores of these strains to different concentrations of the agents for different periods of time. For some of the agents, an exposure of 10 min was required for sporicidal activity. Further, only Actichlor was able to bring about a 3 log(10) reduction in spore numbers under clean and dirty conditions. It was also the only agent that decontaminated different hard, non-porous surfaces artificially contaminated with C. difficile spores. However, this too required an exposure time of more than 2 min and up to 10 min. In conclusion, only the chlorine-releasing agent Actichlor was found to be suitable for the elimination of C. difficile spores from the environment, making it the agent of choice for the decontamination of laboratory surfaces.


The effects of Bt Cry1Ie toxin on bacterial diversity in the midgut of Apis mellifera ligustica (Hymenoptera: Apidae).
The honey bee has been regarded as a key species in the environmental risk assessment of biotech crops. Here, the potential adverse effects of Cry1Ie toxin on the midgut bacteria of the worker bees (Apis mellifera ligustica) were investigated under laboratory conditions. Newly emerged bees were fed with different concentrations of Cry1Ie toxin syrups (20 ng/mL, 200 ng/mL, and 20 μg/mL), pure sugar syrup, and 48 ppb of imidacloprid syrups, then sampled after 15 and 30 d. We characterized the dominant midgut bacteria and compared the composition and structure of the midgut bacterial community in all samples using the Illumina MiSeq platform targeting the V3-V4 regions of 16S rDNA. No significant differences in the diversity of the midgut bacteria were observed between the five treatments. This work was the first to show the effects of Cry1Ie toxin on honey bees, and our study provided a theoretical basis for the biosafety assessment of transgenic Cry1Ie maize.


Multi-omics analyses of early liver injury reveals cell-type-specific transcriptional and epigenomic shift.
Liver fibrosis is a wound-healing response to tissue injury and inflammation hallmarked by the extracellular matrix (ECM) protein deposition in the liver parenchyma and tissue remodelling. Different cell types of the liver are known to play distinct roles in liver injury response. Hepatocytes and liver endothelial cells receive molecular signals indicating tissue injury and activate hepatic stellate cells which produce ECM proteins upon their activation. Despite the growing knowledge on the molecular mechanism underlying hepatic fibrosis in general, the cell-type-specific gene regulatory network associated with the initial response to hepatotoxic injury is still poorly characterized. In this study, we used thioacetamide (TAA) to induce hepatic injury in adult zebrafish. We isolated three major liver cell types - hepatocytes, endothelial cells and hepatic stellate cells - and identified cell-type-specific chromatin accessibility and transcriptional changes in an early stage of liver injury. We found that TAA induced transcriptional shifts in all three cell types hallmarked by significant alterations in the expression of genes related to fatty acid and carbohydrate metabolism, as well as immune response-associated and vascular-specific genes. Interestingly, liver endothelial cells exhibit the most pronounced response to liver injury at the transcriptome and chromatin level, hallmarked by the loss of their angiogenic phenotype. Our results uncovered cell-type-specific transcriptome and epigenome responses to early stage liver injury, which provide valuable insights into understanding the molecular mechanism implicated in the early response of the liver to pro-fibrotic signals.


Superoxide is an associated signal for apoptosis in axonal injury.
Optic neuropathy is the leading cause of irreversible blindness, and a paradigm for central nervous system axonal disease. The primary event is damage to retinal ganglion cell axons, with subsequent death of the cell body by apoptosis. Trials of neuroprotection for these and other neuronal diseases have mostly failed, primarily because mechanisms of neuroprotection in animals do not necessarily translate to humans. We developed a methodology for imaging an intracellular transduction pathway that signals neuronal death in the living animal. Using longitudinal confocal scanning multilaser ophthalmoscopy, we identified the production of superoxide within retrograde-labelled rat retinal ganglion cells after optic nerve transection. Superoxide was visualized by real-time imaging of its reaction product with intravitreally administered hydroethidine and confirmed by differential spectroscopy of the specific product 2-hydroxyethidium. Retinal ganglion cell superoxide increased within 24 h after axotomy, peaking at 4 days, and was not observed in contralateral untransected eyes. The superoxide signal preceded phosphatidylserine externalization, indicating that superoxide generation was an early event and preceded apoptosis. Intravitreal pegylated superoxide dismutase blocked superoxide generation after axotomy and delayed retinal ganglion cell death. Together, these results are consistent with superoxide being an upstream signal for retinal ganglion cell apoptosis after optic nerve injury. Early detection of axonal injury with superoxide could serve as a predictive biomarker for patients with optic neuropathy.


[Recent advances in research on Malassezia microbiota in humans].
Malassezia species of lipophilic yeasts account for most fungal microbiota. Although they colonize healthy skin, they are also associated with several skin diseases, including pityriasis versicolor, seborrheic dermatitis, Malassezia folliculitis, and atopic dermatitis. To date, 14 members of the Malassezia genus have been identified. Of these, both M. globosa and M. restricta predominate, regardless of skin-disease type. Comprehensive analysis of fungal microbiota in the skin of patients with atopic dermatitis using an rRNA clone library method revealed that fungal microbiota cluster according to disease severity. The external ear canal and sole of the foot are colonized by specific Malassezia microbiota.


Microglial inflammation after chronic spinal cord injury is enhanced by reactive astrocytes via the fibronectin/β1 integrin pathway.
After spinal cord injury (SCI), glial scarring is mainly formed around the lesion and inhibits axon regeneration. Recently, we reported that anti-β1 integrin antibody (β1Ab) had a therapeutic effect on astrocytes by preventing the induction of glial scar formation. However, the cellular components within the glial scar are not only astrocytes but also microglia, and whether or not β1Ab treatment has any influence on microglia within the glial scar remains unclear. To evaluate the effects of β1Ab treatment on microglia within the glial scar after SCI, we applied thoracic contusion SCI to C57BL/6N mice, administered β1Ab in the sub-acute phase, and analyzed the injured spinal cords with immunohistochemistry in the chronic phase. To examine the gene expression in microglia and glial scars, we selectively collected microglia with fluorescence-activated cell sorting and isolated the glial scars using laser-captured microdissection (LMD). To examine the interaction between microglia and astrocytes within the glial scar, we stimulated BV-2 microglia with conditioned medium of reactive astrocytes (RACM) in vitro, and the gene expression of TNFα (pro-inflammatory M1 marker) was analyzed via quantitative polymerase chain reaction. We also isolated both naïve astrocytes (NAs) and reactive astrocytes (RAs) with LMD and examined their expression of the ligands for β1 integrin receptors. Statistical analyses were performed using Wilcoxon's rank-sum test. After performing β1Ab treatment, the microglia were scattered within the glial scar and the expression of TNFα in both the microglia and the glial scar were significantly suppressed after SCI. This in vivo alteration was attributed to fibronectin, a ligand of β1 integrin receptors. Furthermore, the microglial expression of TNFα was shown to be regulated by RACM as well as fibronectin in vitro. We also confirmed that fibronectin was secreted by RAs both in vitro and in vivo. These results highlighted the interaction mediated by fibronectin between RAs and microglia within the glial scar. Microglial inflammation was enhanced by RAs via the fibronectin/β1 integrin pathway within the glial scar after SCI. Our results suggested that β1Ab administration had therapeutic potential for ameliorating both glial scar formation and persistent neuroinflammation in the chronic phase after SCI.


Conserved and nonconserved epitopes among Haemophilus influenzae type b pili.
We investigated the binding of antibodies raised against four different Haemophilus influenzae type b (Hib) plus antigen preparations to the native pili and denatured pilins of 21 Hib isolates. Antibodies against live piliated Hib M43p+, adsorbed with a nonpiliated variant to remove nonpilus antibodies, bound to 18 of the 21 piliated Hib isolates in immunodot assays but failed to recognize the denatured pilins from any of the strains in Western immunoblot assays. Similarly, antibodies against purified native pili of strain E1ap+ bound to 11 of 21 piliated strains in immunodot assays but to only 2 of 21 piliated strains in Western blot assays. The native pili of all 21 strains were recognized by one or both of the antisera. These observations suggest that the immunodominant epitopes of native Hib pili are dependent on conformation and are moderately conserved. In contrast, antibodies against denatured M43p+ pilin or against a peptide derived from amino acids 5 through 17 of M43p+ pilin failed to bind to native pili from any of the 21 piliated isolates on immunodot assay. However, both sera recognized the denatured pilins from all the piliated strains on Western blot assay. These data indicate that the immunodominant epitopes of denatured pilins are highly conserved among different strains of Hib but are unavailable on intact pili for antibody binding.


Alpha-adrenergic receptors in platelets from patients with Raynaud's syndrome.
The pathophysiologic basis for spastic Raynaud's syndrome remains undefined but may be related to altered adrenergic activity. The relationship between Raynaud's syndrome and alterations in adrenergic receptor populations was examined in the present study by evaluation of alpha 2-adrenergic receptors in platelets. Direct binding assays revealed 78 +/- 8 fm/mg protein of alpha 2-adrenergic receptors in the platelets from control subjects. Similar levels of alpha 2-adrenergic receptors were observed in patients with obstructive Raynaud's syndrome. In contrast, platelet alpha 2-adrenergic receptor levels were significantly elevated (182 +/- 15 fm/mg protein) in the platelets from patients with spastic Raynaud's syndrome. These results indicate the presence of an altered alpha 2-adrenergic receptor population in the platelets from patients with spastic Raynaud's syndrome, which may be related to the vasospasm experienced by these patients.


Thyroid dysfunction alters gene expression of proteins related to iron homeostasis and metabolomics in male rats.
Thyroid hormones (THs) are crucial in bodily functions, while iron is essential for processes like oxygen transport. Specialized proteins maintain iron balance, including ferritin, transferrin, ferroportin, and hepcidin. Research suggests that THs can influence iron homeostasis by affecting mRNA and protein expression, such as ferritin and transferrin. Our study focused on male rats to assess mRNA expression of iron homeostasis-related proteins and metabolomics in thyroid dysfunction. We found altered gene expression across various tissues (liver, duodenum, spleen, and kidney) and identified disrupted metabolite patterns in thyroid dysfunction. These findings highlight tissue-specific effects of thyroid dysfunction on essential iron homeostasis proteins and provide insights into associated metabolic changes. Our research contributes to understanding the intricate interplay between thyroid hormones and iron balance. By unveiling tissue-specific gene expression alterations and metabolic disruptions caused by thyroid dysfunction, our work lays a foundation for future investigations to explore underlying mechanisms and develop targeted strategies for managing iron-related complications in thyroid disorders.


Magnetic resonance imaging: an accurate, radiation-free, alternative to computed tomography for the primary imaging and three-dimensional reconstruction of the bony orbit.
To determine the extent to which the accuracy of magnetic resonance imaging (MRI) based virtual 3-dimensional (3D) models of the intact orbit can approach that of the gold standard, computed tomography (CT) based models. The goal was to determine whether MRI is a viable alternative to CT scans in patients with isolated orbital fractures and penetrating eye injuries, pediatric patients, and patients requiring multiple scans in whom radiation exposure is ideally limited. Patients who presented with unilateral orbital fractures to the Royal Brisbane and Women's Hospital from March 2011 to March 2012 were recruited to participate in this cross-sectional study. The primary predictor variable was the imaging technique (MRI vs CT). The outcome measurements were orbital volume (primary outcome) and geometric intraorbital surface deviations (secondary outcome) between the MRI- and CT-based 3D models. Eleven subjects (9 male) were enrolled. The patients' mean age was 30 years. On average, the MRI models underestimated the orbital volume of the CT models by 0.50 ± 0.19 cm(3). The average intraorbital surface deviation between the MRI and CT models was 0.34 ± 0.32 mm, with 78 ± 2.7% of the surface within a tolerance of ±0.5 mm. The volumetric differences of the MRI models are comparable to reported results from CT models. The intraorbital MRI surface deviations are smaller than the accepted tolerance for orbital surgical reconstructions. Therefore, the authors believe that MRI is an accurate radiation-free alternative to CT for the primary imaging and 3D reconstruction of the bony orbit.


Psychological and social correlates of doping attitudes among Italian athletes.
This study aims to identify the main psychological and social correlates of doping attitudes among Italian athletes. It is well recognized that athlete disposition and attitude towards doping is one of the factors responsible for doping behavior. Less is known, however, about the factors that sustain the level of athletes' attitudes towards doping. The main psychological (i.e., perfectionism, sport motivation, self-confidence and life satisfaction) and social correlates (i.e., social network and contact with people who use sports drugs) of attitudes towards doping among Italian athletes are examined in this paper. Differences are hypothesized regarding the type of sport (resistance sport vs. non-resistance sport) and athlete participation in competitive sport (i.e., agonistics) or in non-competitive sport (i.e., amateurs) on the level of attitude towards doping. The research hypothesis is that each of these constructs affects the level of athletes' attitudes toward doping. Data were collected from a sample of athletes (N=109), aged from 15 to 45 (M=31.5; SD=13.78) recruited in a Sports Medicine Center. Socio-demographic information, attitude towards doping, psychological and social variables were assessed through self-report questionnaire. Hierarchical multiple regression showed that both psychological (i.e., extrinsic motivation, perfectionism) and social variables (i.e., athletes' contact with doping users) were associated with athletes' attitudes towards doping. The results highlighted that athletes with excessive perfectionism, extrinsically motivated and who have contact with doping users have a positive attitude toward doping. Athletes who exhibit these characteristics should be considered at risk and monitored to prevent possible future sports drug use.


Flange-Based Hand-Eye Calibration Using a 3D Camera With High Resolution, Accuracy, and Frame Rate.
Point cloud data provides three-dimensional (3D) measurement of the geometric details in the physical world, which relies heavily on the quality of the machine vision system. In this paper, we explore the potentials of a 3D scanner of high quality (15 million points per second), accuracy (up to 0.150 mm), and frame rate (up to 20 FPS) during static and dynamic measurements of the robot flange for direct hand-eye calibration and trajectory error tracking. With the availability of high-quality point cloud data, we can exploit the standardized geometric features on the robot flange for 3D measurement, which are directly accessible for hand-eye calibration problems. In the meanwhile, we tested the proposed flange-based calibration methods in a dynamic setting to capture point cloud data in a high frame rate. We found that our proposed method works robustly even in dynamic environments, enabling a versatile hand-eye calibration during motion. Furthermore, capturing high-quality point cloud data in real-time opens new doors for the use of 3D scanners, capable of detecting sensitive anomalies of refined details even in motion trajectories. Codes and sample data of this calibration method is provided at Github (https://github.com/ancorasir/flange_handeye_calibration).


Serum lipoprotein measurement--liquid chromatography and sequential floatation (ultracentrifugation) compared.
An HPLC method [J. Biochem. (Tokyo) 91:1381, 1982] was used for evaluating serum lipoproteins, with on-line monitoring of either cholesterol or phospholipids. Five well-distinguished lipoprotein fractions were observed, based on their particle sizes. Serum of 15 normal persons, 12 subjects with various types of hyperlipidemia, 20 patients with various liver diseases, and two cases of familial LCAT deficiency were examined and the results compared with those by a sequential ultracentrifugal floatation technique. In the normal group, the amounts of fractions 2, 3, and 4 by the HPLC method correlated well with concentrations of the LDL, HDL2, and HDL3 fractions as measured by the ultracentrifugal method, respectively. In the hyperlipidemic group, similar good correlations were observed between fractions 1, 2, 3, 4 and chylomicrons + VLDL, LDL, HDL2, and HDL3 fractions, respectively. For those with liver diseases or LCAT deficiency, the corresponding fractions correlated less well, and characteristically the elution profile of lipoproteins in these groups showed heterogeneity of particle size within each lipoprotein density class, especially in LDL and HDL2.


Low income as a protective factor against asthma in children and adolescents treated via the Brazilian Unified Health System.
To analyze the role that low income plays in the development of asthma in children and adolescents. A case-control study using a questionnaire. A total of 687 participants were studied at a tertiary hospital via the Brazilian Unified Health System. Ages ranged from 5 to 15 years, and 54.7% of the participants were male. Nearly half of the individuals (49.1%) lived in the metropolitan area of the city of Recife, and the remainder lived in the countryside. Most (98.1%) lived in concrete houses or apartments, with a mean of 5.7 rooms and 4.8 occupants per residence. Mean maternal level of education was 6.8 years of schooling. The median monthly per capita income was R$ 103.75 (Brazilian reals). The sample was stratified according to cut-off points related to the national minimum wage (R$350.00): <25% of minimum wage = very low income (39% of the sample); <50% of minimum wage = low income (37.3% of the sample); and > or =50% of minimum wage = satisfactory income (23.7% of the sample). There was no association between poverty and development of asthma. Poverty is not protective against asthma in children and adolescents treated via the Brazilian Unified Health System, as we might suppose based on the hygiene hypothesis. However, cohort studies are needed in order to confirm these findings.


Morphologic and morphometric analysis of testis of Pseudis limellum (Cope, 1862) (Anura, Hylidae) during the reproductive cycle in the Pantanal, Brazil.
The spermatogenesis of Pseudis limellum, from the Southern Pantanal, Brazil, was studied from July 1995 to May 1996, through histological sections of the testis. The cells could be differentiated as: primary spermatogonia, large cells, generally with bilobed nucleus; secondary spermatogonia, smaller cells, with darker cytoplasm, chromatin of radial form; primary and secondary spermatocytes, differentiated according to the different stages of the nucleus during the successive cells divisions. Furthermore, we observed cells in process of morphologic differentiation: rounded spermatids much smaller, with nucleus containing chromatin in compacting process and cytoplasm reduction; elongated spermatids, generally parallel organized in well defined bundles, with the anterior region directed toward the periphery of the seminiferous tubule and the tail directed toward the lumen. Spermatozoa are free in the lumen of the seminiferous tubule. All the cells are organized as cysts, which are supported by a large amount of Sertoli cells. The spermatogenesis in P. limellum is very similar to that of other anurans, but peculiarities were observed regarding the organization of the germ cells, the great amount of free Sertoli cells in the lumen of testis collected in May, and the long cytoplasmatic extensions of the cells bearing pigments and involving the seminiferous tubule. The diameter of the seminiferous tubule (SD) exhibited an annual mean of 251.79 +/- 37.57 microm. Spermatozoa number by seminiferous tubule (SN) exhibited an annual mean of 306.66 +/- 39.83, also with higher and lower values at each month. Variations in SD and SN were not significantly correlated with climatic variables. In this species, reproduction occurs throughout the year in ponds and flooded areas, despite the seasonal climate of the Pantanal. Although males varied in their annual reproductive activity, they were considered potentially reproductive in all months throughout the year.


Changes in histamine synthetic activity, histamine content and responsiveness to compound 48/80 with maturation of rat peritoneal mast cells.
Intact rat peritoneal mast cells were separated by elutriation into fractions differing in cell size, the smallest cells (less than 12 microns) exhibited high histamine synthetic activity (200-300 pmol of histidine decarboxylated per hr/10(6) cells) and contained little histamine (less than 2 pg/cell). With increasing cell size, histamine synthetic activity diminished to less than 20 pmol/hr/10(6) cells and histamine content increased to greater than 12 pg/mast cell. A gradation in histamine release in response to Compound 48/80 was also observed; small mast cells were resistant, those of intermediate size (12-13 microns diameter) were partially responsive and large mast cells (14-17 microns) were fully responsive to Compound 48/80. These changes were related to the state of maturity of the mast cells as indicated by their histochemical reactions to Alcian blue and safranin, which stain nonsulfated heparin precursors and heparin, respectively. Fractions of small mast cells were predominantly immature cells (70-85% of the cells were Stage I) with few granules, whereas fractions of large cells contained mostly mature cells (greater than 80% Stages III and IV) with numerous heparin-containing granules. Fractions in between contained mast cells at intermediate stages (Stages II and III) of development. Degenerative changes, namely decreased viability and diminished responsiveness to Compound 48/80, were evident, however, in fractions of the largest cells (greater than 17 microns). This and other work indicate that, even in adult rats, peritoneal mast cells are heterogeneous and can be separated on the basis of size into subpopulations that differ markedly in histamine content, histamine synthetic activity and ability to release histamine in response to Compound 48/80.


Diet quality, school attendance, and body weight status in adolescent girls in rural Guatemala.
A cluster of factors affects nutritional status among adolescent girls in low- and middle-income countries (LMICs). We investigated the association between school attendance and diet quality among 498 rural adolescent girls (352 attending and 146 not attending school) in Tecpán, Guatemala. In a cross-sectional study, we collected sociodemographic and anthropometric data and characterized the dietary intake using a Food Frequency Questionnaire. We then calculated diet quality using the Healthy Eating Score (HES). Multiple linear regression models were conducted to evaluate the effects of school attendance on diet quality. We found that the overall diet quality among the study participants was poor, according to the HES. However, those who attended school had significantly higher intakes of vitamin A-rich fruits and vegetables (P = 0.04), other fruits (P = 0.01), and milk and milk products (P = 0.004), but a higher intake of fast foods, chips, and saturated fatty acids (P < 0.001). Furthermore, the effects of school attendance on diet quality were significant after adjusting for sociodemographic and lifestyle factors (β coefficient = -1.70, 95% CI: -3.30 to -0.11) but was attenuated when further adjusted for weight status (β coefficient = -1.58, 95% CI: -3.17 to 0.02). Our findings suggest that diet quality among girls in rural Guatemala is poor, particularly among those who do not attend school. To advance our understanding of adolescent diet in LMICs, future studies should include adolescents who are out of school.


Thermal Stresses in Maize: Effects and Management Strategies.
Climate change can decrease the global maize productivity and grain quality. Maize crop requires an optimal temperature for better harvest productivity. A suboptimal temperature at any critical stage for a prolonged duration can negatively affect the growth and yield formation processes. This review discusses the negative impact of temperature extremes (high and low temperatures) on the morpho-physiological, biochemical, and nutritional traits of the maize crop. High temperature stress limits pollen viability and silks receptivity, leading to a significant reduction in seed setting and grain yield. Likewise, severe alterations in growth rate, photosynthesis, dry matter accumulation, cellular membranes, and antioxidant enzyme activities under low temperature collectively limit maize productivity. We also discussed various strategies with practical examples to cope with temperature stresses, including cultural practices, exogenous protectants, breeding climate-smart crops, and molecular genomics approaches. We reviewed that identified quantitative trait loci (QTLs) and genes controlling high- and low temperature stress tolerance in maize could be introgressed into otherwise elite cultivars to develop stress-tolerant cultivars. Genome editing has become a key tool for developing climate-resilient crops. Moreover, challenges to maize crop improvement such as lack of adequate resources for breeding in poor countries, poor communication among the scientists of developing and developed countries, problems in germplasm exchange, and high cost of advanced high-throughput phenotyping systems are discussed. In the end, future perspectives for maize improvement are discussed, which briefly include new breeding technologies such as transgene-free clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas)-mediated genome editing for thermo-stress tolerance in maize.


Temporal increase in D614G mutation of SARS-CoV-2 in the Middle East and North Africa.
Phylogeny construction can help to reveal evolutionary relatedness among molecular sequences. The spike (S) gene of SARS-CoV-2 is the subject of an immune selective pressure which increases the variability in such region. This study aimed to identify mutations in the S gene among SARS-CoV-2 sequences collected in the Middle East and North Africa (MENA), focusing on the D614G mutation, that has a presumed fitness advantage. Another aim was to analyze the S gene sequences phylogenetically. The SARS-CoV-2 S gene sequences collected in the MENA were retrieved from the GISAID public database, together with its metadata. Mutation analysis was conducted in Molecular Evolutionary Genetics Analysis software. Phylogenetic analysis was done using maximum likelihood (ML) and Bayesian methods. A total of 553 MENA sequences were analyzed and the most frequent S gene mutations included: D614G = 435, Q677H = 8, and V6F = 5. A significant increase in the proportion of D614G was noticed from (63.0%) in February 2020, to (98.5%) in June 2020 (p < 0.001). Two large phylogenetic clusters were identified via ML analysis, which showed an evidence of inter-country mixing of sequences, which dated back to February 8, 2020 and March 15, 2020 (median estimates). The mean evolutionary rate for SARS-CoV-2 was about 6.5 × 10-3 substitutions/site/year based on large clusters' Bayesian analyses. The D614G mutation appeared to be taking over the COVID-19 infections in the MENA. Bayesian analysis suggested that SARS-CoV-2 might have been circulating in MENA earlier than previously reported.


Integrative transcriptomic, proteomic, and phosphoproteomic analysis on the defense response to Magnaporthe oryzae reveals different expression patterns at the molecular level of durably resistant rice cultivar Mowanggu.
Rice blast, caused by Magnaporthe oryzae is one of the most destructive diseases of rice (Oryza sativa L.) in most rice-cultivated areas worldwide. Mowanggu (MWG) is a traditional landrace rice variety in Yunnan with broad-spectrum and durable blast resistance against rice blast fungus. However, the underlying disease-resistance mechanisms remain unknown. An integrative transcriptomic, proteomic, and phosphoproteomic analysis of MWG was performed after inoculation with M. oryzae in this study. The transcriptomic and proteomic results revealed that MWG was moderately correlated at the transcriptional and protein levels. Differentially expressed genes and proteins were up-regulated and significantly enriched in protein phosphorylation, peroxisome, plant-pathogen interactions, phenylpropanoid metabolism and phenylalanine biosynthesis pathways. The phosphoproteomic profile and phosphorylated-protein-interaction network revealed that the altered phosphoproteins were primarily associated with reactive oxygen species (ROS), glycolysis, MAPK signaling pathways, and amino acid biosynthesis. In addition, a series of physiological and biochemical parameters, including ROS, soluble sugars, soluble protein and callus accumulation and defense-related enzyme activities, were used to validate the possible blast resistance mechanisms of MWG. The integrative transcriptomic, proteomic, and phosphoproteomic analysis revealed the different expression patterns at the molecular level of the durably resistant rice cultivar MWG after inoculation with M. oryzae, which provides insight into the molecular mechanisms of rice blast resistance.


[The problem of adequate analgesia in the postoperative period].
Despite numerous methods of drug and non-drug analgesia, acute pain relief remains a pressing problem, particularly for the postoperative period. Individual protocols of analgesia are still to be developed. Standard doses of analgesics, administered in some intensive care wards, may be inadequate in some patients. An increase of an opioid dose may lead to untoward reactions. Therefore, we consider therapy with nonsteroid antiinflammatory drugs (ketorolac) justified. An important trend in improvement of the efficacy of drug analgesia is evaluation of a sufficient analgesic dose. Patient-controlled analgesia (PCA), administered in accordance with the patient's request, is an alternative to the traditional analgesia administered according to indications (planned analgesia). The main advantage of PCS in comparison with traditional administration of analgesics are effective analgesia meeting the individual requirements of a patient, rapid desired effect, a shorter period without analgesia, stable concentration of the analgesic in the plasma, time saving for the staff, and a lower incidence of side effects. PCA has been used in 227 patients in intensive care wards of Research Center of Surgery. The majority of patients appreciate this method high, which results in adequate analgesia in 82-95% cases.


[Physical and technical bases of tumor hemostasis with a CO2 laser in neurosurgery].
The particular thermal properties of CO2 laser make it an useful instrument for neurological surgery. Necrosis and carbonization zones, as well as its thermal diffusion in the nervous tissue are dramatically less important with a CO2-laser unit than with other lasers (Nd-Yag or Argon) and moreover with the electrical mono--or bipolar coagulator. That is why the authors chosed this instrumentation for achieving a good hemostasis. It was particularly useful during removal of intracranial tumors situated close to the functional structures of the basis of the brain and the brainstem. The authors present their experience with the french CO2-laser unit Ercelas 80 (Robert et Carrière Biomedical) that they have been using routinely since 1983 in 60 neurosurgical procedures. They present the technical standards necessary to perform such an hemostasis either with the hand-piece or using the micromanipulator. In all cases the output power must be much lower for hemostasis than for tumoral cutting or vaporization. Depending on the type of tumor, radiation power should vary from 2 to 10 watts if a linear emission is used, and from 10 to 30 watts in case of pulsed emission. On the other hand, CO2-laser beam must be unfocussed so as to avoid cutting the vessels less than 0.5 mm in diameter before coagulating them properly.


Disease-free and overall survival at 3.5 years for neoadjuvant bevacizumab added to docetaxel followed by fluorouracil, epirubicin and cyclophosphamide, for women with HER2 negative early breast cancer: ARTemis Trial.
The ARTemis trial previously reported that addition of neoadjuvant bevacizumab (Bev) to docetaxel (D) followed by fluorouracil, epirubicin and cyclophosphamide (D-FEC) in HER2 negative breast cancer improved the pathological complete response (pCR) rate. We present disease-free survival (DFS) and overall survival (OS) with central pathology review. Patients were randomized to 3 cycles of D followed by 3 cycles of FEC (D-FEC), ±4 cycles of Bev (Bev + D-FEC). DFS and OS were analyzed by treatment and by central pathology reviewed pCR and Residual Cancer Burden (RCB) class. A total of 800 patients were randomized [median follow-up 3.5 years (IQR 3.2-4.4)]. DFS and OS were similar across treatment arms [DFS hazard ratio (HR)=1.18 (95% CI 0.89-1.57), P = 0.25; OS HR = 1.26 (95% CI 0.90-1.76), P = 0.19). Both local pathology report review and central histopathology review confirmed a significant improvement in DFS and OS for patients who achieved a pCR [DFS HR = 0.38 (95% CI 0.23-0.63), P < 0.001; OS HR = 0.43 (95% CI 0.24-0.75), P = 0.003]. However, significant heterogeneity was observed (P = 0.02); larger improvements in DFS were obtained with a pCR achieved with D-FEC than a pCR achieved with Bev + D-FEC. As RCB class increased, significantly worse DFS and OS was observed (P for trend <0.0001), which effect was most marked in the ER negative group. The addition of short course neoadjuvant Bev to standard chemotherapy did not demonstrate a DFS or OS benefit. Achieving a pCR with D-FEC is associated with improved DFS and OS but not when pCR is achieved with Bev + D-FEC. At the present time therefore, Bev is not recommended in early breast cancer. NCT01093235.


Clinical features of 36 cases of amelanotic melanomas and considerations about the relationship between histologic subtypes and diagnostic delay.
Amelanotic melanomas (AM) are a difficult diagnostic challenge for clinicians. To consider the clinical presentation of AM, the histologic subtypes involved, the relationship with the diagnostic delay and the possible involvement in overall prognosis. Patients who were observed in our department to be affected by cutaneous melanomas were recorded. Sex, age, the clinical features, the site of presentation, the suspected diagnosis, the clinical course, the histological type, the Clark level and the Breslow thickness were recorded. AM were divided in three main clinical types: an erythematous macule or patch on sun-exposed skin, a dermal plaque or nodule without a particular epidermal change, an exophytic nodule. Only pure AM were considered. Histological subtypes considered were superficial spreading melanoma, nodular melanoma, and lentigo maligna melanoma. Diagnostic delay considered from when the patients first noticed the lesion on the site where the melanoma was diagnosed and when the physician or the patient first proposed the removal was recorded. The chi-squared test was used for statistical evaluation with P < 0.05 as level of significance. Thirty-six cases of AM out of a total of 500 melanomas (7.2%) were collected. The most frequent morphology of clinical presentation was the papulo-nodular form, followed by the plaque form. Mean Breslow thickness of AM was 1.72 mm compared to 0.61 mm of pigmented cases. Nodular histotype was highly represented in AM (30.5% of cases) with respect to pigmented nodular melanomas (2.9%). The diagnostic delay did not differ between amelanotic and pigmented melanomas, nor between nodular AM and nodular pigmented melanomas. The great prevalence of clinical and histological nodular cases, the higher mean Breslow thickness (considered as the most important factor of prognosis) of AM compared with a not significant greater diagnostic delay may point out that a good percentage of AM have an intrinsic faster speed of growth with a worse prognosis irrespectively of the diagnostic performance. The importance of educational campaign for patient and physicians is stressed.


Neuropsychological assessment of patients with late onset GM2 gangliosidosis.
To characterize cognitive status in a sample of individuals with late-onset GM2 gangliosidosis (commonly referred to as late-onset Tay-Sachs disease). Seventeen subjects (13 men, 4 women) diagnosed with GM2 gangliosidosis were evaluated. Subjects ranged in age from 18 to 56 years and were in various stages of disease progression. Subjects underwent comprehensive neuropsychological assessment. Impairment was defined as performance more than 1.6 SD below the normative mean. Group mean performance was within the denoted normal range on all measures except on a task assessing visual sequencing and set shifting. Approximately one-half of the sample scored in the impaired range on measures of processing speed, visual sequencing, and set shifting. One-third of the sample also scored in the impaired range on measures of delayed verbal recall. Impairment tended to be restricted to a subset of the sample, as 5 of the 14 subjects able to undergo formal testing accounted for 70% of the total number of impaired scores. If the three subjects unable to participate in formal testing are also considered impaired, 47% of the current sample exhibited significant cognitive impairment in at least one cognitive domain. In late-onset GM2 gangliosidosis, there is a risk of impairment in executive functioning and memory as well as cerebellar dysfunction. Dementia was not present in any subjects in the current sample.


[Clinical and epidemiologic description of a severe outbreak of Salmonellosis in an urban nursery school].
We describe clinically and epidemiologically an outbreak of gastrointestinal infection by Salmonella enterica ser. (serotype) Enteritidis in an urban infant school, which led to high morbidity and significant social alarm. The immediate communication, as well as the adequate study of the outbreak, in both aspects, allowed identifying the pathogen and establishing control measures in a reasonable period of time. Controversial aspects such as the indication of antibiotherapy or the moment of closing the center are discussed. We retrospectively collected clinical, analytical and epidemiological information and we reviewed the methodology of the outbreak study and its results. A total of 57 children (3-45 months), were affected and had microbiological confirmation. Diarrhea and fever were the main symptoms. 74% went to the hospital and 37% were admitted (mean stay 3.3 days). Factors associated with admission were: dehydration, significant elevation of acute phase reactants and coagulopathy. Twelve patients received parenteral cefotaxime. There were 2 complications: 1 bacteremia and 1 readmission. The initial suspicion of the origin of the outbreak was food, but the analysis of the control samples was negative. Five workers were positive (2 symptomatic). Epidemiologic Surveillance concluded that the probable origin of the outbreak was an asymptomatic carrier and improper diapers handling. The center was closed for 8 days. Cleaning and disinfection measures were carried out, as well as instruction on diaper changing, and the carriers were followed. Clustering in time and space of cases should be reported immediately for early control of the outbreak. Children may present severe forms of Salmonella gastroenteritis.


Suspension and optical properties of the crystalline lens in the eyes of basal vertebrates.
We have investigated the apparatus suspending the crystalline lens in the eyes of basal vertebrates. Data are presented for Holocephali (Chondrichthyes) and the actinopterygians Polypteriformes, Polyodontidae (Acipenseriformes), Lepisosteiformes, Amiiformes, and one teleost species, the banded archerfish (Toxotes jaculatrix). We also studied the optical properties of the lens in Polypteriformes, Lepisosteiformes, and the archerfish. Together with previously published results, our findings show that there are three basic types of lens suspension in vertebrates. These are i) a rotationally symmetric suspension (Petromyzontida, lampreys; Ceratodontiformes, lungfishes; Tetrapoda), ii) a suspension with a dorso-ventral axis of symmetry and a ventral papilla (all Chondrichthyes and Acipenseriformes), and iii) an asymmetric suspension with a ventral muscle and a varying number of ligaments (all Actinopterygii except for Acipenseriformes). Large eyes with presumably high spatial resolution have evolved in all groups. Multifocal lenses creating well-focused color images are also present in all groups studied. Stable and exact positioning of the lens, in many cases in combination with accommodative changes in lens position or shape, is achieved by all three types of lens suspension. It is somewhat surprising that lens suspensions are strikingly similar in Chondrichthyes and Acipenseriformes (Actinopterygii), while the suspension apparatus in Polypteriformes, usually being regarded as an actinopterygian group more basal than Acipenseriformes, are considerably more teleostean-like. This study completes a series of investigations on lens suspensions in nontetrapod vertebrates, covering all major groups except for the rare and highly derived coelacanths.


Rhodococcus kyotonensis sp. nov., a novel actinomycete isolated from soil.
A polyphasic study was undertaken to establish the taxonomic position of an isolate, strain DS472(T), from soil in Kyoto, Japan. Phylogenetic analysis, based on the 16S rRNA gene sequences, revealed that this strain constitutes a new subline within the genus Rhodococcus, with Rhodococcus yunnanensis YIM 70056(T) and Rhodococcus fascians DSM 20669(T) as its nearest phylogenetic neighbours (98.2 and 97.8 % sequence similarity, respectively). DNA-DNA hybridization experiments revealed 36 and 29 % relatedness between the isolate and its phylogenetic relatives, R. yunnanensis and R. fascians, respectively. Chemotaxonomic characteristics, including the major quinone MK-8(H(2)), predominant fatty acids C(16 : 0), C(18 : 1)omega9c and 10-methyl C(18 : 0), the presence of cell-wall chemotype IV and mycolic acids, were consistent with the properties of members of the genus Rhodococcus. The DNA G+C content was 64.5 mol%. On the basis of both phenotypic and genotypic evidence, strain DS472(T) represents a novel species of the genus Rhodococcus, for which the name Rhodococcus kyotonensis sp. nov. is proposed. The type strain is strain DS472(T) (=IAM 15415(T)=CCTCC AB206088(T)).


Rictor is a novel target of p70 S6 kinase-1.
The rapamycin-insensitive companion of mammalian target of rapamycin (mTOR) (Rictor) is a key member of mTOR complex-2 (mTORC2), which phosphorylates the AGC kinases Akt/PKB, PKC and SGK1 at a C-terminal hydrophobic motif. We identified several novel sites on Rictor that are phosphorylated, including Thr1135, which is conserved across all vertebrates. Phosphorylation of this site on Rictor is stimulated by amino acids and growth factors through a rapamycin-sensitive signaling cascade. We demonstrate here that Rictor is a direct target of the ribosomal protein S6 kinase-1 (S6K1). Rictor phosphorylation at Thr1135 does not lead to major changes in mTORC2-kinase activity. However, phosphorylation of this site turns over rapidly and mediates 14-3-3 binding to Rictor and mTORC2, providing possibility for altered interactions of the complex. These findings reveal an unexpected signaling input into mTORC2, which is regulated by amino acids, growth factors and rapamycin.


An EEG/ERP investigation of the development of empathy in early and middle childhood.
Empathic arousal is the first ontogenetic building block of empathy to appear during infancy and early childhood. As development progresses, empathic arousal becomes associated with an increasing ability to differentiate between self and other, which is a critical aspect of mature empathetic ability (Decety and Jackson, 2004). This allows for better regulation of contagious distress and understanding others mental states. In the current study, we recorded electroencephalographic event-related potentials and mu suppression induced by short visual animations that depicted painful situations in 57 typically developing children aged between 3 and 9 years as well as 15 young adults. Results indicate that the difference wave of an early automatic component (N200), indexing empathic arousal, showed an age-related decrease in amplitude. In contrast, the difference wave of late-positive potentials (LPP), associated with cognitive appraisal, showed an age-related gain. Only early LPP was detected in children, whereas both early and late LPP were observed in adults. Furthermore, as compared with adults, children showed stronger mu suppression when viewing both painful and non-painful stimuli. These findings provide neurophysiological support for the development of empathy during childhood, as indicated by a gradual decrease in emotional arousal and an increase in cognitive appraisal with age.


Kinase activity, heat shock protein 27 phosphorylation, and lung epithelial cell glutathione.
The 27-kDa heat shock protein (Hps27) is phosphorylated in a way that appears to regulate antioxidant defenses by mitogen-activated protein kinase (MAPK)-activated protein kinase 2 (MK2), a component of the p38(MAPK) pathway. To investigate the role of Hsp27 in cellular resistance to oxidant stress, lung cells (A549) were incubated with MAPK inhibitors to investigate the pathway's role in antioxidant defense. Cells were harvested for measurement of reduced gluthathione and glutathione disulfide (GSH and GSSH); or, exposed to 2,3-dimethoxy-1,4-napthoquinone (DMNQ). Inhibition of MAPK with SB203580 decreased total cellular glutathione (mean +/- SE): Vehicle, 150 +/- 20 mu M; SB203580, 57 +/- 10* (*P < .01). Inhibition of MAPK tripled [GSSG]/[GSH]: Vehicle, 0.29 +/- 0.09; SB203580, 1.06 +/- 0.43* (*P > .05; n = 6 per group). Hsp27 protein content did not change significantly after MAPK inhibition: Vehicle 2.20 +/- 0.24 ng/mg protein; SB203580, 2.03 +/- 0.34 (P > .05). Transfection of epithelial cells with wild-type (pcDNA-HA-Hsp27) or phosphomimic (pcDNA-HA-Hsp27-S3D) vector increased Hsp27 protein, which significantly protected cells from oxidant stress. Inhibition of the MAPK system, including p38(MAPK), results in cellular oxidant stress. Hsp27, which is phosphorylated by MK2 in the MAPK pathway, protects epithelial cells from oxidant stress.


Novel clues on abiotic stress tolerance emerge from embryo proteome analyses of rice varieties with contrasting stress adaptation.
Cereal embryos sustain severe water deficit at the final stage of seed maturation. The molecular mechanisms underlying the acquisition of desiccation tolerance in seed embryos are similar to those displayed during water deficit in vegetative tissues. The genetic variation among six rice genotypes adapted to diverse environmental conditions was analysed at the proteome level to get further clues on the mechanisms leading to water-stress tolerance. MS analysis allowed the identification of 28 proteins involved in stress tolerance (late embryogenesis abundant proteins), nutrient reservoir activity, among other proteins implicated in diverse cellular processes potentially related to the stress response (e.g., mitochondrial import translocase). Hierarchical clustering and multidimensional scaling analyses revealed a close relationship between the stress-sensitive genotypes, whereas the stress-tolerant varieties were more distantly related. Besides qualitative and significant quantitative changes in embryo proteins across the distinct varieties, we also found differences at post-translational level. The results indicated that late embryogenesis abundant Rab21 was more strongly phosphorylated in the embryos of the sensitive varieties than in the embryos of the tolerant ones. We propose that the differences found in the phosphorylation status of Rab21 are related to stress tolerance.


Studies on the molecular recognition between bioactive peptides and angiotensin-converting enzyme.
High blood pressure or hypertension is a condition affecting many individuals and represents a controllable risk factor for cardiovascular diseases such as coronary heart disease and stroke. A non-pharmacological approach to manage these includes the application of food components with antihypertensive activity. Milk protein-derived peptides have been exploited as natural hypotensive agents, namely the peptides Val-Pro-Pro (VPP) and Ile-Pro-Pro (IPP), already commercialized in functional foods as a potential alternative to synthetic drugs. These bioactive peptides inhibit in vitro and in vivo the Angiotensin I-converting enzyme (ACE), a protein with an important role in blood pressure regulation. In this work, we attempted to elucidate the possible mode of interaction between the peptides and ACE, including mechanisms of binding to the cofactor Zn2+, and further contrast this with the known mode of inhibition exerted by synthetic drugs (Captopril, Enalaprilat and Lisinopril). The bioactive peptide Ala-Leu-Pro-Met-His-Ile-Arg (ALPMHIR), also known to inhibit the enzyme ACE but with a lower efficiency than VPP and IPP, was utilized in the docking studies for comparison. It was observed that the best docking poses obtained for VPP and IPP were located at the ACE catalytic site with very high resemblance to the drugs mode of interaction, including the coordination with Zn2+. As for ALPMHIR, the best docking poses were located in the narrow ACE channel outside the catalytic site, representing higher affinity energies and fewer resemblances with the interaction established by drugs.


A late, solitary brain metastasis of epithelial ovarian carcinoma.
Brain metastasis from epithelial ovarian cancer (EOC) is very rare with a reported incidence of less than 2%. It is usually associated with a poor prognosis that is related to several factors, the most important including: single vs multiple lesions, performance status, platinum-sensitive disease, tumor grade, extracranial disease, and multimodal approach treatment. At the time of diagnosis, an extracranial disease is found in over half of patients. The most common histology is the serous type. The median time from primary diagnosis to development of cerebral lesions is directly correlated to initial tumor grade and stage. Several therapeutic approaches can be proposed, including best supportive care +/- corticosteroids, surgery, radiotherapy and chemotherapy. A multimodal therapy approach may achieve an improved outcome and should therefore be utilized whenever applicable. We present the case of a patient with a solitary brain metastasis which appeared 11 years after a locally advanced and aggressive EOC (FIGO stage III C) and which totally regressed after surgery and adjuvant chemotherapy. Clinically, she showed progressive headaches, decreased visual acuity, balance and memory disorders associated with a confusional state. Brain CT scan and MRI documented a solitary, necrotic lesion in the left central parietal region with an important cerebral surrounding edema and initial cranial herniation. No other extracranial metastases were observed at the PET scan. Laboratory tests were in the normal range and CA 125 was moderatly increased at 81 UI/ml. The patient underwent surgical removal of tumor lesion, post-surgical whole-brain radiotherapy (WBRT) and systemic chemotherapy with carboplatin alone for six cycles. At a follow-up of 13 months, she is alive, in good clinical condition and tumor progression free. The peculiarity of this case relies on the isolated brain relapse of a BRCA-1/BRCA-2 non-mutated EOC, which is uncommon and rare, and to the very long time, of 11 years, from diagnosis of primary cancer and development of brain metastasis. A multimodal, aggressive approach of this isolated brain metastasis led to a complete and prolonged tumor control.


Fast minimization of structural risk by nearest neighbor rule.
In this paper, we present a novel nearest neighbor rule-based implementation of the structural risk minimization principle to address a generic classification problem. We propose a fast reference set thinning algorithm on the training data set similar to a support vector machine (SVM) approach. We then show that the nearest neighbor rule based on the reduced set implements the structural risk minimization principle, in a manner which does not involve selection of a convenient feature space. Simulation results on real data indicate that this method significantly reduces the computational cost of the conventional SVMs, and achieves a nearly comparable test error performance.


The Fabrication of a Probe-Integrated Electrochemiluminescence Aptasensor Based on Double-Layered Nanochannel Array with Opposite Charges for the Sensitive Determination of C-Reactive Protein.
The effective and sensitive detection of the important biomarker, C-reactive protein (CRP), is of great significance in clinical diagnosis. The development of a convenient and highly sensitive electrochemiluminescence (ECL) aptasensor with an immobilized emitter probe is highly desirable. In this work, a probe-integrated ECL aptamer sensor was constructed based on a bipolar silica nanochannel film (bp-SNF) modified electrode for the highly sensitive ECL detection of CRP. The bp-SNF, modified on an ITO electrode, consisted of a dual-layered SNF film, including the negatively charged inner SNF (n-SNF) and the outer SNF with a positive charge and amino groups (p-SNF). The ECL emitter, tris(bipyridine) ruthenium (II) (Ru(bpy)32+), was stably immobilized in a nanochannel of bp-SNF using the dual electrostatic interactions with n-SNF attracting and p-SNF repelling. The amino groups on the outer surface of bp-SNF were aldehyde derivatized, allowing for the covalent immobilization of recognitive aptamers (5'-NH2-CGAAGGGGATTCGAGGGGTGATTGCGTGCTCCATTTGGTG-3'), leading to the recognition interface. When CRP bound to the aptamer on the recognition interface, the formed complex increased the interface resistance and reduced the diffusion of the co-reactant tripropylamine (TPA) into the nanochannels, leading to a decrease in the ECL signal. Based on this mechanism, the constructed aptamer sensor could achieve a sensitive ECL detection of CRP ranging from 0.01 to 1000 ng/mL, with a detection limit (DL) of 8.5 pg/mL. The method for constructing this probe-integrated ECL aptamer sensor is simple, and it offers a high probe stability, good selectivity, and high sensitivity.


The time-dependent reconstructed evolutionary process with a key-role for mass-extinction events.
The homogeneous reconstructed evolutionary process is a birth-death process without observed extinct lineages. Each species evolves independently with the same diversification rate-speciation rate, λ(t), and extinction rate, μ(t)-that may change over time. The process is commonly applied to model species diversification where the data are reconstructed phylogenies, e.g. trees estimated from present-day molecular data, and used to infer diversification rates. In the present paper I develop the general probability density of a reconstructed tree under any homogeneous, time-dependent birth-death process. I demonstrate how to adapt this probability density when conditioning on the survival of one or two initial lineages, or on the process realizing n species, and also how to transform between the probability density of a reconstructed tree and the probability density of the speciation times. I demonstrate the use of the general time-dependent probability density functions by deriving the probability density of a reconstructed tree under a birth-death-shift model with explicit mass-extinction events. I extend these functions to several special cases, including the pure-birth process, the pure-death process, the birth-death process, and the critical-branching process. Thus, I specify equations for the most commonly used birth-death models in a unified framework (e.g. same condition and same data) using a common notation.


Comparison of risk factor reduction and tolerability of a full-dose polypill (with potassium) versus low-dose polypill (polycap) in individuals at high risk of cardiovascular diseases: the Second Indian Polycap Study (TIPS-2) investigators.
A daily single capsule (polycap) of 3 blood pressure (BP) lowering drugs (hydrochlorthiazide, 12.5 mg; atenolol, 50 mg; ramipril, 5 mg) at low doses, simvastatin (20 mg), and aspirin (100 mg) has been demonstrated to be well tolerated and to reduce BP and low-density lipoprotein cholesterol. We examined the incremental effects of 2 (full dose) plus K(+) supplementation versus single polycap (low dose) on risk factors and tolerability. After a run-in period, 518 individuals with previous vascular disease or diabetes mellitus from 27 centers in India were randomly assigned to a single-dose polycap or to 2 capsules of the polycap plus K(+) supplementation for 8 weeks. The effects on BP, heart rate (HR), serum lipids, serum and urinary K(+), and tolerability were assessed using an intention-to-treat analysis. The full-dose polycap (plus K(+) supplementation) reduced BP by a further 2.8 mm Hg systolic and 1.7 mm Hg diastolic, compared with that observed with the low-dose polycap (P=0.003; P=0.001), but there were no differences in HR (0.1 bpm). The differences in total and low-density lipoprotein cholesterol between the full-dose and low-dose polycap was 7.2 mg/dL (P=0.014) and 6.6 mg/dL (P=0.006), respectively, but there were no differences in high-density lipoprotein cholesterol or triglycerides. The rates of discontinuation of the study drug after randomization were similar in the 2 groups (6.9% low dose versus 7.8% full dose). The full-dose polycap (plus K(+) supplementation) reduces BP and low-density lipoprotein cholesterol to a greater extent compared with the low dose, with similar tolerability. Therefore, the full-dose polycap should potentially lead to larger benefits. Clinical Trial Registration- URL: http://www.ctri.nic.in. Unique identifier: CTRI/2010/091/000054.