The effect of valproic acid in combination with irradiation and temozolomide on primary human glioblastoma cells.
Glioblastoma multiforme (GBM) has nearly uniformly fatal with a median survival of less than 2 years. While there have not been any novel anti-GBM therapeutics approved for many years, there has been the gradual accumulation of clinical data suggesting that the widely used anti-convulsant agent, valproic acid (VPA) may significantly prolong survival in GBM patients. This pre-clinical study aimed to determine the potential clinical utility of VPA in the treatment of GBM. Primary GBM cells were treated with VPA as a monotherapy and in combination with temozolomide and irradiation. At clinically achievable concentrations, VPA was shown to be effective as a monotherapy agent in the five primary lines tested. VPA was then used as a sensitizing agent to in vitro radiation and showed significant augmentation of in vitro irradiation therapy. In addition, when VPA, radiation and temozolomide were combined an additive, rather than synergistic effect was noted. Gene expression profiling demonstrated close clustering of triple treated cells with VPA mono-treated cells while untreated cells clustered closer with TMZ-irradiation dual treated cells. These microarray data suggest a dominant role of VPA at the gene expression level when combining these different treatment options. Moreover, in an in vivo tumor transplantation model, we were able to demonstrate an increase in animal survival when cells were pre-treated with irradiation-VPA and when triple treated. These findings provide a significant rationale for the investigation of VPA in the treatment of GBM patients.


Popliteal artery repair in massively transfused military trauma casualties: a pursuit to save life and limb.
Popliteal artery war wounds can bleed severely and historically have high rates of amputation associated with ligation (72%) and repair (32%). More than before, casualties are now surviving the initial medical evacuation and presenting with severely injured limbs that prompt immediate limb salvage decisions in the midst of life-saving maneuvers. A modern analysis of current results may show important changes because previous limb salvage strategies were limited by the resuscitation and surgical techniques of their eras. Because exact comparisons between wars are difficult, the objective of this study was to calculate a worst-case (a pulseless, fractured limb with massive hemorrhage from popliteal artery injury) amputation-free survival rate for the most severely wounded soldiers undergoing immediate reconstruction to save both life and limb. We performed a retrospective study of trauma casualties admitted to the combat support hospital at Ibn Sina Hospital in Baghdad, Iraq, between 2003 and 2007. US military casualties requiring a massive transfusion (> or = 10 blood units transfused within 24 hours of injury) were identified. We extracted data on the subset of casualties with a penetrating supra or infrageniculate popliteal arterial vascular injury. Demographics, injury mechanism, Injury Severity Score, tourniquet use, physiologic parameters, damage control adjuncts, surgical repair techniques, operative time, and outcomes (all-cause 30-day mortality, amputation rates, limb salvage failure, and graft patency) were investigated. Forty-six massively transfused male casualties, median age 24 years (range, 19-54 years; mean Injury Severity Score, 19 +/- 8.0), underwent immediate orthopedic stabilization and vascular reconstruction. There was one early death. The median operative time for the vascular repairs was 217 minutes (range, 94-630 minutes) and included all damage control procedures. Combined arterial and venous injuries occurred in 17 (37%). Ligation was performed for no arterial and 9 venous injuries. Amputations (transtibial or transfemoral) were considered limb salvage failures (14 of 48, 29.2%) and were grouped as immediate (< or = 48 hours, 5), early (>48 hours and < or = 30 days, 6), or late (>30 days, 3). Limb losses were from graft thrombosis, infection, or chronic pain. Combined arterial and venous injuries occurred in 17 (37%). Ligation was performed for no arterial and nine venous injuries. For a median follow-up (excluding death) of 48 months (range, 23-75 months), the amputation-free survival rate was 67%. This study, a worst-case study, showed comparable results to historical controls regarding limb salvage rates (71% for Iraq vs. 56-69% for the Vietnam War). Thirty-day survival (98%), 4-year amputation-free survival (67%), and complication-free rates (35%) fill knowledge gaps. Guidelines for managing popliteal artery injuries show promising results because current resuscitation practices and surgical care yielded similar amputation rates to prior conflicts despite more severe injuries. Significant transfusion requirements and injury severity may not indicate a life-over-limb strategy for popliteal arterial repairs. Future studies of limb salvage failures may help improve casualty care by reducing the complications that directly impact amputation-free survival.


Accuracy of screening for diabetic retinopathy by family physicians.
We wanted to examine the accuracy of family physicians' screening for diabetic retinopathy using standardized criteria and a nonmydriatic ophthalmoscope. Eleven family physicians assessed 28 standardized patients with diabetes mellitus using the PanOptic ophthalmoscope. Their assessments of whether the patients required referral to an ophthalmologist were compared with the reference standard of retinal diagrams. The mean sensitivity for the family physicians was 87% (95% confidence interval [CI], 83%-91%) with a specificity of 57% (95% CI, 46%-68%). Overall agreement was moderate, with a mean kappa = .43 (95% CI, 0.39%-0.47%). Using standardized criteria and a nonmydriatic ophthalmoscope, family physicians were fairly accurate in screening patients for diabetic retinopathy. Whereas this technique is not sufficiently accurate to replace routine referral for all patients with diabetes, it can be used to improve care for those patients who fail to get routine eye screenings.


Effects of zinc on intestinal injury and some serum parameters in ethanol-administered rats.
This study was designed to determine the morphological and biochemical effects of zinc sulfate and the role of metallothionein in ethanol-induced intestinal injury. Rats received zinc sulfate (100 mg/kg/d) for 3 consecutive d, 2 h prior to the administration of ethanol by gavage. Ethanol administration caused intestinal injury as determined by increased serum lactate dehydrogenase activity, urea, creatinine, uric acid, and sialic acid levels, intestinal lipid peroxidation level, decreased serum catalase activity, intestinal glutathione level, and metallothionein expression. Zinc sulfate pretreatment of the ethanol group caused a decrease in histological damage, serum lactate dehydrogenase activity, urea, creatinine, uric acid, sialic acid levels, and intestinal lipid peroxidation level, but increases in serum catalase activity, intestinal glutathione level, and metallothionein expression. The present study indicates that zinc sulfate has a protective effect against ethanol-induced intestinal injury. In addition, the protective effect of zinc on ethanol-induced intestinal injury might be mediated by metallothionein, as well as having antioxidative potential.


Health-related quality of life of immune thrombocytopenic purpura patients: results from a web-based survey.
To assess the health-related quality of life (HRQOL) of immune thrombocytopenic purpura (ITP) patients. This was a cross-sectional, descriptive study comparing ITP patients' HRQOL to age and gender matched controls. ITP patients from the Platelet Disorder Support Association were recruited until 1000 surveys had been completed. Controls were randomly sampled from the Harris Interactive Online Panel. ITP patients and controls completed a one-time web-based survey, including a comprehensive HRQOL assessment. ITP patients completed the SF-36, the EQ-5D, and the ITP-Patient Assessment Questionnaire (ITP-PAQ). Controls completed the SF-36 and EQ-5D only. ITP patients' SF-36 and EQ-5D scores were compared to controls in unadjusted and adjusted analyses. Associations between splenectomy status, duration of illness, and platelet count with ITP patients' HRQOL scores were also examined. This analysis included 1002 ITP patients and 1031 controls. ITP patients scored worse on seven of eight SF-36 domains and the Physical and Mental Summary scores (all p < 0.05) and on the EQ-5D visual analog scale (65.5 vs. 82.3; p = 0.002). ITP patients who had undergone splenectomy had similar SF-36 and EQ-5D scores to non-splenectomy patients but scored significantly worse on 5 of 10 ITP-PAQ scales: Bother, Psychological, Fear, Social Activity, and Work (all p < 0.05). ITP patients diagnosed within the past 5 years had worse Bother and Overall Quality of Life scores than less recently diagnosed patients but were similar on other ITP-PAQ scales. Lower platelet count was consistently associated with worse ITP-PAQ scores and had weaker associations with SF-36 and EQ-5D scores. ITP was associated with consistent and statistically significant deficits on generic HRQOL measures. The ITP-PAQ demonstrated differences based on disease severity and treatments. The self-selection bias in the two samples limits the generalizability of the results to all patients with ITP. Further research is needed in more generalizable samples.


Activation of murine CD4+ and CD8+ T lymphocytes leads to dramatic remodeling of N-linked glycans.
Differentiation and activation of lymphocytes are documented to result in changes in glycosylation associated with biologically important consequences. In this report, we have systematically examined global changes in N-linked glycosylation following activation of murine CD4 T cells, CD8 T cells, and B cells by MALDI-TOF mass spectrometry profiling, and investigated the molecular basis for those changes by assessing alterations in the expression of glycan transferase genes. Surprisingly, the major change observed in activated CD4 and CD8 T cells was a dramatic reduction of sialylated biantennary N-glycans carrying the terminal NeuGcalpha2-6Gal sequence, and a corresponding increase in glycans carrying the Galalpha1-3Gal sequence. This change was accounted for by a decrease in the expression of the sialyltransferase ST6Gal I, and an increase in the expression of the galactosyltransferase, alpha1-3GalT. Conversely, in B cells no change in terminal sialylation of N-linked glycans was evident, and the expression of the same two glycosyltransferases was increased and decreased, respectively. The results have implications for differential recognition of activated and unactivated T cells by dendritic cells and B cells expressing glycan-binding proteins that recognize terminal sequences of N-linked glycans.


Deep learning-based phenotyping reclassifies combined hepatocellular-cholangiocarcinoma.
Primary liver cancer arises either from hepatocytic or biliary lineage cells, giving rise to hepatocellular carcinoma (HCC) or intrahepatic cholangiocarcinoma (ICCA). Combined hepatocellular- cholangiocarcinomas (cHCC-CCA) exhibit equivocal or mixed features of both, causing diagnostic uncertainty and difficulty in determining proper management. Here, we perform a comprehensive deep learning-based phenotyping of multiple cohorts of patients. We show that deep learning can reproduce the diagnosis of HCC vs. CCA with a high performance. We analyze a series of 405 cHCC-CCA patients and demonstrate that the model can reclassify the tumors as HCC or ICCA, and that the predictions are consistent with clinical outcomes, genetic alterations and in situ spatial gene expression profiling. This type of approach could improve treatment decisions and ultimately clinical outcome for patients with rare and biphenotypic cancers such as cHCC-CCA.


Drospirenone--a new progestogen with antimineralocorticoid activity, resembling natural progesterone.
In the second half of a normal menstrual cycle, progesterone levels rise. Progesterone binds to its specific receptor, but also to the mineralocorticoid receptor; thus progesterone acts as a mineralocorticoid antagonist. For this reason, natriuresis is slightly enhanced in the luteal phase, and, as a reflection of the negative sodium balance, plasma renin and aldosterone rise by 20-50%. This rise is of a compensatory nature, and prevents further sodium losses. All conventional synthetic progestogens, whether they are derivatives of 17alpha-hydroxyprogesterone or 19-nortestosterone, lack the antimineralocorticoid effect of natural progesterone. Ethinylestradiol, as the estrogenic component of combined oral contraceptives, is a sodium-retaining drug. This effect is mainly due to a significant increase of the hepatic synthesis of renin substrate (angiotensinogen). Even with low-dose oral contraceptives, systolic and diastolic blood pressure may be raised in susceptible individuals. Drospirenone is a new progestogen, derived from 17alpha-spirolactone, and the relationship between its progestogenic and its antimineralocorticoid potency is almost identical to that of natural progesterone. In an early preclinical study in 12 normal young women, it was found that the oral administration of 2 mg drospirenone for 6 days led to a cumulative sodium loss of 84 mmol and a significant rise in plasma renin and aldosterone, compared with placebo. In a second experiment, it was found that 2 mg drospirenone given from cycle days 5 to 25 to six regularly menstruating women suppressed ovulation and led to a slight natriuresis without a change in blood pressure, while renin and aldosterone levels slightly increased. The natriuresis and the increase in renin and aldosterone levels did not occur in six other women who received 1 mg cyproterone acetate (a progestogen with antiandrogenic properties), instead of drospirenone. Consequently, an oral contraceptive was designed containing 30 microg ethinylestradiol and 3 mg drospirenone (Yasmin, Schering AG, Berlin, Germany) in the hope of developing a contraceptive that might prevent the sodium retention brought about by the effect of ethinylestradiol. In a 6-month study involving 20 regularly menstruating women, it was shown that the addition of drospirenone to ethinylestradiol did indeed prevent the small rise in body weight and blood pressure observed in some women taking a conventional oral contraceptive. In all studies conducted so far with Yasmin, cycle control and tolerability have been found to be good. In conclusion, Yasmin may become an especially well-tolerated combined oral contraceptive, due to the striking similarity between its progestogenic component drospirenone and progesterone.


Elevated Choroidal Thickness and Central Serous Chorioretinopathy in the Fellow Eyes of Patients with Circumscribed Choroidal Hemangioma.
To determine whether increased choroidal thickness (CT) is associated with circumscribed choroidal hemangioma (CCH) and whether patients with CCH are predisposed to central serous chorioretinopathy (CSCR). We performed a retrospective consecutive observational study of 15 fellow eyes of 15 patients who had CCH. A- and B-scan ultrasonography, fluorescein angiography (FA), and optical coherence tomography (OCT) findings were reviewed to confirm the diagnosis of CCH. Fellow eye CT was measured using enhanced depth imaging OCT. FA, indocyanine green angiography, and OCT images of fellow eyes were reviewed for signs of CSCR. Mean subfoveal CT was significantly greater in the fellow eyes of patients with CCH than in age-matched normal eyes (p = 0.004). Three of 15 (20.0%) fellow eyes of CCH patients had retinal pigment epithelium disruptions and/or pigment epithelial detachments indicative of previous asymptomatic CSCR. In this pilot study, elevated CT was associated with a risk of developing CCH as well as CSCR in patients of varying ages. Patients diagnosed with CCH should be screened for CSCR in the fellow eye. Further exploration of this association may reveal useful clues about the biology of abnormally elevated choroidal hyperpermeability and its various clinical manifestations.


Greater decrements in neuromuscular function following interval compared to continuous eccentric cycling.
Our aim was to determine the demands and consequences of a single session of continuous (CONT) or interval (INT) eccentric cycling. Fourteen healthy males performed 'work-matched' CONT and INT eccentric cycling in a cross over design. Measures of maximal voluntary contraction (MVC), resting twitch force, voluntary activation (VA), muscle soreness and creatine kinase (CK) were taken at baseline, immediately post, and 24, 48 and 72 h post the first exercise bout. The second bout was used to characterise within session demands. Decreases in MVC (INT 19%, CONT 13%), twitch force (INT 31%, CONT 18%), and VA (INT 10%, CONT 6%) were observed immediately post session (p < 0.05). Reductions in twitch force were greater after INT (p < 0.05) and lasted 48 h. Muscle soreness was greater following INT, versus CONT (p < 0.05), although no differences in CK were observed. Metabolic demands (% of V̇O2peak and [BLa]) were greater during INT vs. CONT (32 ± 6% 28 ± 6%; p < 0.001), [BLa] (1.0 ± 0.4 vs. 0.8 ± 0.2 mmol·L-1; p < 0.001) and RPE (12 ± 1 vs. 11 ± 1; p < 0.001), respectively. Total time under tension was 48% greater in CONT compared to INT (p < 0.001), whereas average torque (during exercise) was 40% greater during INT compared to CONT (p < 0.001). Interval eccentric cycling exacerbates muscle soreness, decrements in muscle function and lengthens recovery compared to a work matched continuous bout, which is attributable to increased force rather than time under tension.


Dimethylnitrosamine demethylation by reconstituted liver microsomal cytochrome P-450 enzyme system.
Oxidative demethylation of dimethylnitosamine was studied with both reconstituted and unresolved liver microsomal cytochrome P-450 enzyme systems from rats and hamsters. Proteinase treatment of liver microsomal preparations yielded cytochrome P-450 particulate fractions. Both cytochrome P-450 and NADPH- cytochrome c reductase fractions were required for optimum demethylation activity. Particulate cytochrome P-450 fractions were more effecient than either Triton X-100- or cholatesolubilized preparations of these particles in demethylation activity with rat and hamster liver preparations appear to be due to differences in specificity in their cytochrome P-450 fractions.


Thioredoxin-1 binds to the C2 domain of PTEN inhibiting PTEN's lipid phosphatase activity and membrane binding: a mechanism for the functional loss of PTEN's tumor suppressor activity.
Thioredoxin-1 (Trx-1) is a 12 kDa redox protein that is overexpressed in a large number of human tumors. Elevated Trx-1 is associated with increased tumor cell proliferation, inhibited apoptosis, aggressive tumor growth, and decreased patient survival. The molecular mechanisms for the promotion of tumorigenesis by Trx-1 are not known. PTEN is a major tumor suppressor of human cancer that acts by hydrolyzing membrane phosphatidylinositol (PtdIns)-3-phosphates, thus, preventing the activation of the survival signaling kinase Akt by PtdIns-3-kinase. We show that Trx-1 binds in a redox dependent manner to PTEN to inhibit its PtdIns-3-phosphatase activity which results in increased Akt activation in cells. Molecular docking and site-specific mutation studies show that the binding of Trx-1 to PTEN occurs through a disulfide bond between the active site Cys(32) of Trx-1 and Cys(212) of the C2 domain of PTEN leading to steric interference by bound Trx-1 of the catalytic site of PTEN and of the C2 lipid membrane-binding domain. The results of the study suggest that the increased levels of Trx-1 in human tumors could lead to functional inhibition of PTEN tumor suppressor activity providing an additional mechanism for tumorigenesis with loss of PTEN activity.


Predictors of partying prior to bar attendance and subsequent BrAC.
Pre-drinking (e.g., pre-gaming, pre-loading) occurs frequently among young adult and college-aged drinkers and is associated with increased intoxication as well as alcohol-related problems. The purpose of the following study was to examine pre-drinking behaviors in situ, and to test whether drinking intentions mediate the relationship between heavy episodic drinking history and pre-drinking behaviors. We randomly selected a sample of 1040 young adults bar patrons at 32 bars in a major metropolitan city in Southern California and asked participants to complete an interview concerning drinking behavior as well as provide a breath sample used to measure breath alcohol concentration. We used multilevel path analysis to test two meditational models. Results showed drinking intentions mediated the relationship between heavy episodic drinking history and pre-drinking behavior as well as the relationship between heavy episodic drinking history and level of pre-drinking intoxication. Gender did not moderate these meditational relationships suggesting similar pre-drinking behaviors for both men and women. Potential methods to prevent pre-drinking behaviors in this population of young people are discussed.


The general surgery job market: analysis of current demand for general surgeons and their specialized skills.
The majority of general surgery residents pursue fellowships. However, the relative demand for general surgical skills vs more specialization is not understood. Our objective was to describe the current job market for general surgeons and compare the skills required by the market with those of graduating trainees. Positions for board eligible/certified general surgeons in Oregon and Wisconsin from 2011 to 2012 were identified by review of job postings and telephone calls to hospitals, private practice groups, and physician recruiters. Data were gathered on each job to determine if fellowship training or specialized skills were required, preferred, or not requested. Information on resident pursuit of fellowship training was obtained from all residency programs within the represented states. Of 71 general surgery positions available, 34% of positions required fellowship training. Rural positions made up 46% of available jobs. Thirty-five percent of positions were in nonacademic metropolitan settings and 17% were in academic metropolitan settings. Fellowship training was required or preferred for 18%, 28%, and 92% of rural, nonacademic, and academic metropolitan positions, respectively. From 2008 to 2012, 67% of general surgery residents pursued fellowship training. Most general surgery residents pursue fellowship despite the fact that the majority of available jobs do not require fellowship training. The motivation for fellowship training is unclear, but residency programs should tailor training to the skills needed by the market with the goal of improving access to general surgical services.


Assessing the effect of mHealth on child feeding practice in African countries: systematic and meta-analysis.
Poor child feeding practice is a public health problem in Africa. Mobile health (mHealth) is a supportive intervention to improve this problem; however, the evidence available in the current literature is inconsistent and inconclusive in Africa. Some studies state that exclusive breastfeeding is not different between controls and mHealth interventions in the first month. Other studies state that health providers need additional training for the success of mHealth interventions. This systematic review and meta-analysis aims to provide the summarized effect of mHealth on child-feeding practices in Africa to improve future planning and decisions. We conducted a systematic review and meta-analysis based on the published and unpublished evidence gathered from PubMed, Web of Science, Cochrane Library, and Embase databases between January 1, 2000, and March 1, 2022. Studies included were randomized control trials and experimental studies that compared mHealth to standards of care among postpartum women. Preferred Reporting Items for Systematic Review and Meta-analysis guidelines followed for the reporting. After screening 1188 studies, we identified six studies that fulfilled the study criteria. These studies had 2913 participants with the number of total intervention groups 1627 [1627/2913 = 56%]. Five studies were completed within 24 weeks while one required 12 weeks. We included two RCTs, two cluster RCTs, and two quasi-experimental studies all used mHealth as the major intervention and usual care as controls. We found significant improvement in child-feeding practices among intervention groups. This systematic review and meta-analysis showed that the application of mHealth improved child-feeding practices in Africa. Although the finding is compelling, the authors recommend high-quality studies and mHealth interventions that consider sample size, design, regional differences, and environmental constraints to enhance policy decisions. The place of residence, access, low socioeconomic development, poor socio-demographic characteristics, low women empowerment, and low women's education might cause high heterogeneity in the included regions and need consideration during interventions. PROSPERO: CRD42022346950.


Contrasting treatment-specific survival using double-robust estimators.
In settings where a randomized trial is infeasible, observational data are frequently used to compare treatment-specific survival. The average causal effect (ACE) can be used to make inference regarding treatment policies on patient populations, and a valid ACE estimator must account for imbalances with respect to treatment-specific covariate distributions. One method through which the ACE on survival can be estimated involves appropriately averaging over Cox-regression-based fitted survival functions. A second available method balances the treatment-specific covariate distributions through inverse probability of treatment weighting and then contrasts weighted nonparametric survival function estimators. Because both methods have their advantages and disadvantages, we propose methods that essentially combine both estimators. The proposed methods are double robust, in the sense that they are consistent if at least one of the two working regression models (i.e., logistic model for treatment and Cox model for death hazard) is correct. The proposed methods involve estimating the ACE with respect to restricted mean survival time, defined as the area under the survival curve up to some prespecified time point. We derive and evaluate asymptotic results through simulation. We apply the proposed methods to estimate the ACE of donation-after-cardiac-death kidney transplantation with the use of data obtained from multiple centers in the Netherlands.


Criminal conviction among offspring with parental history of mental disorder.
Offspring of parents with mental disorder are at risk of a range of adverse outcomes. We sought to establish whether such risks extend to offending by examining rates of criminal conviction, including conviction for violent and sexual offences, among offspring of parents with mental disorder compared to offspring without parental disorder. From a random sample of the Danish population, a cohort aged ≤15 years (n=412,117) was followed for the occurrence of conviction between January 1981 and December 2006. Incidence rate ratios (IRRs) and cumulative incidences for offspring conviction by parental mental disorder status were calculated using a Cox regression model. Analyses were repeated for conviction for a serious first offence. Offspring with history of parental mental disorder had higher rates of conviction than those without parental disorder; rates were highest for those with two affected parents [IRR 3.39, 95% confidence interval (CI) 3.08-3.73]. The association persisted when parental gender, offspring gender and the nature of parental disorder were considered. Absolute rates were lower but relative rates higher for female offspring (IRR 3.26 for males with two affected parents, 4.52 for females). Similar patterns were seen for conviction for serious offences. Associations were attenuated after adjustment was made for family socio-economic position (SEP) and parental criminality. Offspring of parents with mental disorder represent a group at elevated risk of criminality. This raises the possibility of shared familial vulnerability for mental disorder and criminal behaviour, and highlights the need to consider early identification and intervention in this group.


Evaluation of VDRL test in Sudanese blood donors.
2201 blood donors and 199 patients with sexually transmitted diseases (STD) were tested with the Venereal Disease Research Laboratory (VDRL) and the fluorescent treponemal antibody-absorption (FTA-ABS) tests. The VDRL test was found positive in 30 (1.36%) of the blood donors and 11 (5.5%) of the patients. Syphilis was confirmed by FTA-ABS test in one patient (0.5%) but not in blood donors. The biological false positive (BFP) reactions were 1.36% in blood donors and 5.0% in control patients. The place of VDRL test as a screening test for syphilis and the low rate of its seroreactivity among blood donors was discussed. The BFP reactions were reviewed to assist in the interpretation of the VDRL test which was evaluated on the basis of the control specimens. It was concluded that syphilis should be excluded before blood is transfused and the VDRL test should remain the screening test of choice in the blood transfusion service.


Localization and characterization of flavivirus envelope glycoprotein cross-reactive epitopes.
The flavivirus E glycoprotein, the primary antigen that induces protective immunity, is essential for membrane fusion and mediates binding to cellular receptors. Human flavivirus infections stimulate virus species-specific as well as flavivirus cross-reactive immune responses. Flavivirus cross-reactive antibodies in human sera create a serious problem for serodiagnosis, especially for secondary flavivirus infections, due to the difficulty of differentiating primary from secondary cross-reactive serum antibodies. The presence of subneutralizing levels of flavivirus cross-reactive serum antibodies may result in a dramatic increase in the severity of secondary flavivirus infections via antibody-dependent enhancement. An understanding of flavivirus E-glycoprotein cross-reactive epitopes is therefore critical for improving public health responses to these serious diseases. We identified six E-glycoprotein residues that are incorporated into three distinct flavivirus cross-reactive epitopes. Two of these epitopes which are recognized by distinct monoclonal antibodies contain overlapping continuous residues located within the highly conserved fusion peptide. The third epitope consists of discontinuous residues that are structurally related to the strictly conserved tryptophan at dengue virus serotype 2 E-glycoprotein position 231.


Dramatic Conformer-Dependent Reactivity of the Acetaldehyde Oxide Criegee Intermediate with Dimethylamine Via a 1,2-Insertion Mechanism.
The reactivity of carbonyl oxides has previously been shown to exhibit strong conformer and substituent dependencies. Through a combination of synchrotron-multiplexed photoionization mass spectrometry experiments (298 K and 4 Torr) and high-level theory [CCSD(T)-F12/cc-pVTZ-F12//B2PLYP-D3/cc-pVTZ with an added CCSDT(Q) correction], we explore the conformer dependence of the reaction of acetaldehyde oxide (CH3CHOO) with dimethylamine (DMA). The experimental data support the theoretically predicted 1,2-insertion mechanism and the formation of an amine-functionalized hydroperoxide reaction product. Tunable-vacuum ultraviolet photoionization probing of anti- or anti- + syn-CH3CHOO reveals a strong conformer dependence of the title reaction. The rate coefficient of DMA with anti-CH3CHOO is predicted to exceed that for the reaction with syn-CH3CHOO by a factor of ∼34,000, which is attributed to submerged barrier (syn) versus barrierless (anti) mechanisms for energetically downhill reactions.


Study of chromatic adaptation using memory color matches, Part I: neutral illuminants.
Twelve corresponding color data sets have been obtained using the long-term memory colors of familiar objects as target stimuli. Data were collected for familiar objects with neutral, red, yellow, green and blue hues under 4 approximately neutral illumination conditions on or near the blackbody locus. The advantages of the memory color matching method are discussed in light of other more traditional asymmetric matching techniques. Results were compared to eight corresponding color data sets available in literature. The corresponding color data was used to test several linear (von Kries, RLAB, etc.) and nonlinear (Hunt & Nayatani) chromatic adaptation transforms (CAT). It was found that a simple two-step von Kries, whereby the degree of adaptation D is optimized to minimize the DEu'v' prediction errors, outperformed all other tested models for both memory color and literature corresponding color sets, whereby prediction errors were lower for the memory color sets. The predictive errors were substantially smaller than the standard uncertainty on the average observer and were comparable to what are considered just-noticeable-differences in the CIE u'v' chromaticity diagram, supporting the use of memory color based internal references to study chromatic adaptation mechanisms.


Multiclass screening in urine by comprehensive two-dimensional liquid chromatography time of flight mass spectrometry for residues of sulphonamides, beta-agonists and steroids.
Nowadays routine residue monitoring involves the analysis of many compounds from different classes, mainly in urine. In the past two decades, developments heavily focused on the use of mass spectrometers (MS) and faster and more sensitive MS detectors have reached the market. However, chromatographic separation (CS) was rather ignored and the cognate developments in CS were not in line. As a result, residue analysis did not improve to the extent anticipated. CS by LC x LC is a promising technique and will enable a further increase in the range of compounds and compound classes that can be detected in a single run. In the present study, a self-built LC x LC system, using a 10 port valve, was connected to a single quadrupole MS with electrospray interface. Standards containing a mixture of sulphonamides, β-agonists and (steroid) hormones, 53 compounds, in total, were analysed. Results demonstrated that these compounds were well separated and could be detected at low levels in urine, i.e. limit of detection (LOD) from 1 µg L-1 for most β-agonists to 10 µg L-1 for some sulphonamides and most hormones. To enhance the sensitivity, optimisation was performed on an advanced commercial LC x LC system connected to a full scan accurate MS. This ultimately resulted in a fast high throughput untargeted method, including a simple sample clean-up in a 96-well format, for the analysis of urine samples.


Apolipoprotein E gene polymorphisms are strong predictors of inflammation and dyslipidemia in rheumatoid arthritis.
Rheumatoid arthritis (RA), a condition with a strong genetic etiology, is associated with excess cardiovascular disease (CVD). Dyslipidemia in RA may be driven by inflammation and genetic factors. Apolipoprotein E (ApoE) is important for the regulation of lipid levels and CVD risk and immune function in the general population. We compared the frequency of 2 ApoE single-nucleotide polymorphisms (SNP) in patients with RA and controls, and studied the relationship of ApoE genotypes with lipids and inflammation in RA. A total of 387 patients with well-characterized RA and 420 non-RA controls were studied. Two ApoE SNP, rs7412 (ApoE2) and rs429358 (ApoE4), were identified. Genotypic (p = 0.908) and allelic (p = 0.894) frequencies did not differ between RA and controls. Within RA, the E2 allele was associated with the lowest and E4 allele with the highest levels of total cholesterol (p = 0.007), low-density lipoproteins (p = 0.004), and apolipoprotein B (p = 0.009). The E4 allele was also associated with lower C-reactive protein (p = 0.007), erythrocyte sedimentation rate (p = 0.001), and Disease Activity Score (p = 0.015) compared to the E3 allele. E2 or E4 alleles were not associated with CVD in RA, although a trend was observed (p = 0.074). The frequency of ApoE polymorphisms did not differ between patients with RA and controls. ApoE genotypes are strongly linked to inflammation and lipid levels in RA, raising interest in the prognostic implications of ApoE genotypes.


Close genetic similarity between two sympatric species of tephritid fruit fly reproductively isolated by mating time.
Two sibling species of tephritid fruit fly, Bactrocera tryoni and B. neohumeralis, occur sympatrically throughout the range of B. neohumeralis in Australia. Isolation between the two species appears to be maintained by a difference in mating time: B. tryoni mates at dusk, whereas B. neohumeralis mates during the middle of the day. A morphological difference in humeral callus color also distinguishes the two species. Despite clear phenotypic evidence that B. tryoni and B. neohumeralis are distinct species, genetic differentiation as measured by four markers--nuclear DNA sequences from the white gene and the ribosomal internal transcribed spacer (ITS2), and mitochondrial DNA sequences from the cytochrome b (cytb) and cytochrome oxidase subunit II (COII) genes--is very small. Minor fixed differences occur in the ITS2 sequence, however, in all other cases the two species exhibit a high level of shared polymorphic variation. The close genetic similarity suggests either that speciation has occurred very rapidly and recently in the absence of any mitochondrial DNA sorting or that the sharing of polymorphisms is due to hybridization or introgression. A third species within the tryoni complex, B. aquilonis, is geographically isolated. Bactrocera aquilonis is also genetically very similar, but in this case there is clear differentiation for the mitochondrial loci. The three species form a group of considerable interest for investigation of speciation mechanisms.


Motor simulation and the bodily self.
Previous studies demonstrated the human ability to implicitly recognize their own body. When submitted to a visual matching task, participants showed the so-called self-advantage, that is, a better performance with self rather than others' body or body parts. Here, we investigated whether the body self-advantage relies upon a motor representation of one's body. Participants were submitted to a laterality judgment of self and others' hands (Experiment 1 and 3), which involves a sensory-motor mental simulation. Moreover, to investigate whether the self-advantage emerges also when an explicit self processing is required, the same participants were submitted to an explicit self-body recognition task (Experiment 2). Participants showed the self-advantage when performing the laterality judgment, but not when self-recognition was explicitly required. Thus, implicit and explicit recognition of the bodily self dissociate and only an implicit recognition of the bodily self, mapped in motor terms, allows the self-advantage to emerge.


Effects of antihypertensive drug treatment on the risk of dementia and cognitive impairment.
Dementia is a common and serious health problem that affects 33 million persons globally. With the increase in life expectancy, the prevalence of dementia is expected to reach 81.1 million persons by 2040. Dementia impairs quality of life and is associated with profound disease burden, morbidity, and mortality in both patients and caregivers. Therefore, identifying measures to prevent dementia is a research priority. Midlife hypertension has increased the risk of dementia in large prospective cohort studies. Researchers have investigated the blood pressure-lowering effects of antihypertensive drugs on the incidence of dementia. Although prospective cohort studies have shown that use of antihypertensive drugs was associated with a reduced rate of cognitive impairment and dementia, these studies were not placebo controlled. Four randomized, placebo-controlled studies-the Systolic Hypertension in Europe (Syst-Eur) study, Study on Cognition and Prognosis in the Elderly (SCOPE), Systolic Hypertension in the Elderly Program (SHEP), and Perindopril Protection Against Recurrent Stroke Study (PROGRESS)-investigated the effects of antihypertensive agents on the incidence of dementia. The Syst-Eur study found that active treatment with nitrendipine, enalapril, and/or hydrochlorothiazide reduced the rate of dementia by 50% compared with placebo (p=0.05). The PROGRESS study showed that active treatment with perindopril and indapamide was associated with reduced cognitive decline compared with placebo (risk ratio 19%, p=0.01). In contrast, the SCOPE study (candesartan or hydrochlorothiazide vs placebo) and the SHEP trial (chlorthalidone, atenolol, or reserpine vs placebo) found no significant difference between the active treatment and placebo groups on the incidence of dementia. Some researchers have suggested that certain antihypertensive drug classes, such as angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers, diuretics, and calcium channel blockers, may offer benefit in reducing dementia risk in addition to their blood pressure-lowering effect. Further prospective randomized studies comparing different antihypertensive classes are needed to provide more evidence regarding the effects of antihypertensive drugs on dementia risk and to determine whether certain antihypertensive classes provide greater benefits than others.


Iron acquisition by phytosiderophores contributes to cadmium tolerance.
Based on the ability of phytosiderophores to chelate other heavy metals besides iron (Fe), phytosiderophores were suggested to prevent graminaceous plants from cadmium (Cd) toxicity. To assess interactions between Cd and phytosiderophore-mediated Fe acquisition, maize (Zea mays) plants were grown hydroponically under limiting Fe supply. Exposure to Cd decreased uptake rates of 59Fe(III)-phytosiderophores and enhanced the expression of the Fe-phytosiderophore transporter gene ZmYS1 in roots as well as the release of the phytosiderophore 2'-deoxymugineic acid (DMA) from roots under Fe deficiency. However, DMA hardly mobilized Cd from soil or from a Cd-loaded resin in comparison to the synthetic chelators diaminetriaminepentaacetic acid and HEDTA. While nano-electrospray-high resolution mass spectrometry revealed the formation of an intact Cd(II)-DMA complex in aqueous solutions, competition studies with Fe(III) and zinc(II) showed that the formed Cd(II)-DMA complex was weak. Unlike HEDTA, DMA did not protect yeast (Saccharomyces cerevisiae) cells from Cd toxicity but improved yeast growth in the presence of Cd when yeast cells expressed ZmYS1. When supplied with Fe-DMA as a Fe source, transgenic Arabidopsis (Arabidopsis thaliana) plants expressing a cauliflower mosaic virus 35S-ZmYS1 gene construct showed less growth depression than wild-type plants in response to Cd. These results indicate that inhibition of ZmYS1-mediated Fe-DMA transport by Cd is not related to Cd-DMA complex formation and that Cd-induced phytosiderophore release cannot protect maize plants from Cd toxicity. Instead, phytosiderophore-mediated Fe acquisition can improve Fe uptake in the presence of Cd and thereby provides an advantage under Cd stress relative to Fe acquisition via ferrous Fe.


Glutamatergic postsynaptic density in early life stress programming: Topographic gene expression of mGlu5 receptors and Homer proteins.
Type-5 metabotropic glutamate receptors (mGlu5) have been implicated in the mechanism of resilience to stress. They form part of the postsynaptic density (PSD), a thickening of the glutamatergic synapse that acts as a multimodal hub for multiple cellular signaling. Perinatal stress in rats triggers alterations that make adult offspring less resilient to stress. In the present study, we examined the expression of gene encoding the mGlu5 (Grm5), as well as those encoding the short and long isoforms of Homer proteins in different brain regions of the offspring of dams exposed to repeated episodes of restraint stress during pregnancy ("perinatally stressed" or PRS offspring). To this end, we investigated unconditioned behavioral response using the light/dark box test, as well as the expression of PSD genes (Homer1a, Homer1b, and Grm5), in the medial prefrontal cortex, cortex, caudate-putamen, amygdala, and dorsal hippocampus. PRS rats spent significantly less time in the light area than the control group. In the amygdala, Homer1a mRNA levels were significantly increased in PRS rats, whereas Homer1b and Grm5 mRNA levels were reduced. In contrast, the transcript encoding for Homer1a was significantly reduced in the medial prefrontal cortex, caudate-putamen, and dorsal hippocampus of PRS rats. We also evaluated the relative ratio between Homer1a and Homer1b/Grm5 expression, finding a significant shift toward the expression of Homer1a in the amygdala and toward Homer1b/Grm5 in the other brain regions. These topographic patterns of Homer1a, Homer1b, and mGlu5 gene expression were significantly correlated with risk-taking behavior measured in the light/dark box test. Remarkably, in the amygdala and in other brain regions, Homer1b and Grm5 expression showed positive correlation with time spent in the light box, whereas Homer1a in the amygdala showed a negative correlation with risk-taking behavior, in contrast with all other brain regions analyzed, wherein these correlations were positive. These results suggest that perinatal stress programs the developmental expression of PSD molecules involved in mGlu5 signaling in discrete brain regions, with a predominant role for the amygdala.


Surviving chromosome replication: the many roles of the S-phase checkpoint pathway.
Checkpoints were originally identified as signalling pathways that delay mitosis in response to DNA damage or defects in chromosome replication, allowing time for DNA repair to occur. The ATR (ataxia- and rad-related) and ATM (ataxia-mutated) protein kinases are recruited to defective replication forks or to sites of DNA damage, and are thought to initiate the DNA damage response in all eukaryotes. In addition to delaying cell cycle progression, however, the S-phase checkpoint pathway also controls chromosome replication and DNA repair pathways in a highly complex fashion, in order to preserve genome integrity. Much of our understanding of this regulation has come from studies of yeasts, in which the best-characterized targets are the stimulation of ribonucleotide reductase activity by multiple mechanisms, and the inhibition of new initiation events at later origins of DNA replication. In addition, however, the S-phase checkpoint also plays a more enigmatic and apparently critical role in preserving the functional integrity of defective replication forks, by mechanisms that are still understood poorly. This review considers some of the key experiments that have led to our current understanding of this highly complex pathway.


A genomic analysis of Drosophila somatic sexual differentiation and its regulation.
In virtually all animals, males and females are morphologically, physiologically and behaviorally distinct. Using cDNA microarrays representing one-third of Drosophila genes to identify genes expressed sex-differentially in somatic tissues, we performed an expression analysis on adult males and females that: (1) were wild type; (2) lacked a germline; or (3) were mutant for sex-determination regulatory genes. Statistical analysis identified 63 genes sex-differentially expressed in the soma, 20 of which have been confirmed by RNA blots thus far. In situ hybridization experiments with 11 of these genes showed they were sex-differentially expressed only in internal genital organs. The nature of the products these genes encode provides insight into the molecular physiology of these reproductive tissues. Analysis of the regulation of these genes revealed that their adult expression patterns are specified by the sex hierarchy during development, and that doublesex probably functions in diverse ways to set their activities.


Educational needs assessment for pediatric health care providers on pesticide toxicity.
This pilot project, carried out under the National Strategies for Health Care Providers: Pesticides Initiative, assessed the attitudes, beliefs and practices of pediatricians, nurse practitioners, physicians assistants, and nurses in the metropolitan Washington, D.C., area and the surrounding rural counties regarding health effects of pesticide toxicity and continuing education on pesticide toxicity in the years 2001-2002. Data were collected from practitioners (physicians, physician assistants, and nurse practitioners) and nurses using questionnaires as well as from practitioners using focus groups. Sites for questionnaire distribution and focus groups were selected to represent a variety of practice types and geographic settings. One-hundred-sixty questionnaires from practitioners and 43 from nurses were analyzed. These issues were probed further in six focus groups with 29 participants. Most respondents in both groups did not frequently diagnose or ask questions about pesticide toxicity on patient histories. Most focus group participants were more comfortable answering questions about acute pesticide toxicity, and many relied on poison control centers for assistance with management of acute cases. They expressed less understanding and more uncertainties about chronic toxicity. When asked questions by patients, 64% of practitioners and 69% of nurses felt poorly prepared to answer them. Forty percent of practitioners but only 26% of nurses felt it was important to obtain more information on pesticides. There were divergent preferences on ways to obtain continuing medical education (CME) in general, but a recurrent theme was the need to make CME on pesticide toxicity clinically relevant and one topic among several in a CME conference. Lectures and short courses were the most commonly preferred modes of education among both practitioners and nurses. Educational materials to reach this population of pediatric clinicians on pesticides, as well as other environmental health topics, should make the case justifying the importance of the topics, highlight information of clinical relevance, and use a variety of media. These results should be confirmed before being generalized to a broader group of clinicians, although the consistency of findings between focus groups suggests they are robust, at least for this geographic area.


Low plasma eicosapentaenoic acid and depressive symptomatology are independent predictors of dementia risk.
The potential preventive role of polyunsaturated fatty acids (PUFAs) in Alzheimer disease has aroused increasing interest. Plasma n-3 PUFAs have been shown to be inversely related to the risk of dementia and to depression, which is frequently associated with dementia. The objective was to ascertain whether plasma PUFAs predict the risk of incident dementia in a cohort of older persons, independently of their depressive status. Of 1214 nondemented participants in the Three-City Study from Bordeaux (France) who were followed up for 4 y, 65 developed dementia. The association between the proportion of plasma fatty acids at baseline and the risk of incident dementia was assessed by multivariate proportional hazard models, taking into account depressive status assessed on the basis of the Center for Epidemiologic Studies Depression scale. A higher plasma eicosapentaenoic acid (EPA) concentration was associated with a lower incidence of dementia [hazard ratio (HR) for 1 SD = 0.69; 95% CI: 0.48, 0.98], independently of depressive status and after adjustment for age, education, apolipoprotein E epsilon4 allele, diabetes, and baseline plasma vitamin E and triacylglycerol. The relations between docosahexaenoic acid (DHA), total n-3 PUFAs, and incident dementia did not remain significant in multivariate models. Higher ratios of arachidonic acid (AA) to DHA and of n-6 to n-3 fatty acids were related to an increased risk of dementia, particularly in depressive subjects (n = 90): ratio of AA to DHA (HR: 2.65; 95% CI: 1.07, 6.56) and ratio of n-6 to n-3 (HR: 1.61; 95% CI: 1.04, 2.47). A high plasma EPA concentration may decrease the risk of dementia, whereas high ratios of n-6 to n-3 fatty acids and of AA to DHA may increase the risk of dementia, especially in depressed older persons. The role of EPA in dementia warrants further research.


Current modulated volume-of-interest imaging for kilovoltage intrafaction monitoring of the prostate.
The focus of this work was to improve the available kV image quality for continuous intrafraction monitoring of the prostate during volumetric modulated arc therapy. This is investigated using a novel blade collimation system enabling tube current modulated (TCM) volume-of-interest (VOI) imaging of prostate fiducial markers during radiotherapy, and Monte Carlo simulation of MV scatter. A four-blade dynamic kV collimator was used to track a VOI containing gold fiducial markers embedded in a dynamic pelvis phantom during gantry rotation. For each fiducial, a VOI margin around each marker was set to be 2σ of the population covariance matrix characterizing prostate motion. This was used to conform to a single or several fiducials and compared to a static field. DRRs were used to calculate the kV attenuation for each VOI as a function of angle and used to optimize x-ray tube current during acquisition. Image quality was assessed with regard to contrast-to-noise ratio (CNR), fiducial detectability, and imaging dose. Monte Carlo simulations in EGSnrc were used to calculate the imaging dose to the phantom and MV scatter fluence to the imaging panel. Fiducials can be accurately located using a VOI containing a single or several fiducials using a relatively high constant kV output. However, when using a 6 × 6 cm2 field the dose can be upwards of 1.5 Gy in bone for constant kV output and 3.1 Gy when applying TCM at 1 Hz imaging over the course of 40 fractions. This can be mitigated through tailoring the imaging field to a single or several fiducials, in which the integral dose is reduced by a factor of 15.6 and 3.7, respectively. For a constant MV treatment field size, the scattered fluence reaching the kV panel varies by less than a factor of two for a completely rotation of the gantry. However, the MV scatter spectrum overlaps with the detector response for a deleterious effect, with a peak MV scatter energy of approximately 100 keV. TCM can be used to overcome the variability in image quality throughout the rotation and therefore improve fiducial CNR and detectability during periods of high kV attenuation. The combination of VOI and TCM introduces an advantageous approach in intrafraction monitoring of the prostate during radiotherapy by both reducing and localizing the imaging dose, while improving image quality and fiducial detectability during periods of high kV attenuation. In addition, the influence of MV scatter has been shown to be most important in low attenuation regions, with a variation by a factor of two.


Semaphorin-Mediated Corticospinal Axon Elimination Depends on the Activity-Induced Bax/Bak-Caspase Pathway.
Axon guidance molecules and neuronal activity have been implicated in the establishment and refinement of neural circuits during development. It is unclear, however, whether these guidance molecule- and activity-dependent mechanisms interact with one another to shape neural circuit formation. The formation of corticospinal (CS) circuits, which are essential for voluntary movements, involves both guidance molecule- and activity-dependent components during development. We previously showed that semaphorin6D (Sema6D)-plexinA1 (PlexA1) signaling eliminates ipsilateral projections of CS neurons in the spinal cord, while other studies demonstrate that CS projections to the spinal cord are eliminated in an activity-dependent manner. Here we show that inhibition of cortical neurons during postnatal development causes defects in elimination of ipsilateral CS projections in mice. We further show that mice that lack the activity-dependent Bax/Bak pathway or caspase-9 similarly exhibit defects in elimination of ipsilateral CS projections, suggesting that the activity-dependent Bax/Bak-caspase-9 pathway is essential for the removal of ipsilateral CS projections. Interestingly, either inhibition of neuronal activity in the cortex or deletion of Bax/Bak in mice causes a reduction in PlexA1 protein expression in corticospinal neurons. Finally, intracortical microstimulation induces activation of only contralateral forelimb muscles in control mice, whereas it induces activation of both contralateral and ipsilateral muscles in mice with cortical inhibition, suggesting that the ipsilaterally projecting CS axons that have been maintained in mice with cortical inhibition form functional connections. Together, these results provide evidence of a potential link between the repellent signaling of Sema6D-PlexA1 and neuronal activity to regulate axon elimination.SIGNIFICANCE STATEMENT Both axon guidance molecules and neuronal activity regulate axon elimination to refine neuronal circuits during development. However, the degree to which these mechanisms operate independently or cooperatively to guide network generation is unclear. Here, we show that neuronal activity-driven Bax/Bak-caspase signaling induces expression of the PlexA1 receptor for the repellent Sema6D molecule in corticospinal neurons (CSNs). This cascade eliminates ipsilateral projections of CSNs in the spinal cord during early postnatal development. The absence of PlexA1, neuronal activity, Bax and Bak, or caspase-9 leads to the maintenance of ipsilateral projections of CSNs, which can form functional connections with spinal neurons. Together, these studies reveal how the Sema6D-PlexA1 signaling and neuronal activity may play a cooperative role in refining CS axonal projections.


Magnetoencephalographic spike sources associated with auditory auras in paediatric localisation-related epilepsy.
To characterise magnetoencephalographic spike sources in paediatric patients with auditory auras and recurrent localisation-related epilepsy. Six patients (four boys and two girls (ages 7-14 years) were retrospectively studied. All patients had auditory auras as part of their initial seizure manifestation, including four patients who underwent previous brain surgery. Scalp video electroencephalography and magnetoencephalography (MEG) were carried out in six patients, intraoperative electrocorticography in three patients and extraoperative intracranial video electroencephalography in one patient. MEG auditory-evoked fields (AEFs) were studied in four patients. Three patients had elementary auditory auras, one had complex auditory aura and two had both complex and elementary auras. All six patients had clustered MEG spike sources with coexisting scattered spike sources. MEG clusters were localised in the superior temporal gyrus with surrounding scatters in four patients (two left and two right); two patients had scattered spikes in the superior temporal gyrus in addition to clustered MEG spike sources in the left inferior and middle frontal gyri or parieto-occipital region. AEFs were located within an MEG cluster in one patient and within 3 cm of a cluster in two patients. Surgical resection, including the regions of MEG clusters, was carried out in four patients. Three of four patients who had previous surgeries were seizure free at 2 years after excision of the MEG cluster region. MEG spike sources clustered in the superior temporal gyrus in six patients with auditory auras. These spike sources were in close proximity or seemed to engulf the magnetic AEF. Areas with MEG spike sources contained the residual or recurrent epileptogenic zone after incomplete cortical excision for lesional epilepsy.


[A path analysis on factors influencing second primary cancer screening practices in stomach, colon, and breast cancer survivors].
This study was conducted to identify the factors influencing second primary cancer (SPC) screening practice by examining the relationships of physical symptoms, knowledge and attitudes regarding SPC screening, perceived risk, primary cancer type, and demographic factors of cancer survivors. Participants were 308 survivors of stomach, colon, or breast cancer recruited from 2 university hospitals in Korea. Data were collected using a questionnaire and analyzed using IBM SPSS 21.0 and AMOS 18.0. The proportion of participants taking all cancer screenings according to national guidelines was 40%. They had moderate knowledge and a relatively positive attitude regarding SPC screening and high cancer risk perception. The participants had taken fewer SPC screenings after than before cancer diagnosis. The factors influencing cancer risk perception were age, physical symptoms, knowledge regarding SPC and primary cancer type (stomach). The factors influencing SPC screening practice were age, gender, economic status, knowledge regarding SPC screening, and primary cancer types (colon). It is important for clinical professionals to recognize that survivors of cancer are susceptible to another cancer. Education on SPC screening for these survivors should focus on communicating with and encouraging them to have regular cancer screenings.


Typhoid vaccination for international travelers from Greece visiting developing countries.
Typhoid fever is one of the most common diagnoses in returned international travelers. Our aim was to study the typhoid vaccine prescription practices for travelers from Greece visiting developing countries. A prospective questionnaire-based study was conducted during 2009-2012 in 57 Public Health Departments, which are the only sources of typhoid vaccine in Greece. A total of 3,680 travelers were studied (median age: 38.1 years). Typhoid vaccine was delivered to 1,108 (30.1%) of them. Of those who traveled to sub-Saharan Africa, South America, the Middle East, the Indian subcontinent, Southeast Asia, South Africa, East Asia, North Africa, and Central America, 31.6, 17.1, 35, 44.2, 36.9, 31, 17.7, 31.6, and 36.8% received typhoid vaccine, respectively. Of travelers who stayed <1 month, 1 to <3 months, 3 to <6 months, and ≥6 months, 21.4, 63.1, 32.3, and 34.9% were vaccinated, respectively. According to the purpose of travel, typhoid vaccine was administered to 32.7% of those who traveled for leisure, to 28.8% of those who traveled for business, and to 24.1% of those visiting friends and relatives (VFRs). Of travelers who stayed in urban areas, rural areas, and urban and rural areas, 36.3, 30.1, and 26.8% were vaccinated, respectively. The majority of travelers who received the typhoid vaccine stayed in camps (62.9%) or at local residences (41%). Typhoid vaccine administration was statistically significantly associated with destination, duration of travel, purpose of travel, area of stay, and type of accommodation. There is a need to increase awareness of travelers and public health professionals for typhoid vaccination and particularly for high-risk groups of travelers, such as travelers to the Indian subcontinent and VFRs. Strategies for continuing professional education should be developed for travel health professionals.


On the Thermodynamic Condition for Adsorption Azeotropes.
Adsorption azeotropy is a common phenomenon in mixed-gas adsorption equilibria. Since the first literature report of binary adsorption azeotropes in 1933, numerous studies investigated the thermodynamic conditions for adsorption azeotropes but failed to reach definitive conclusions. Based on the generalized Langmuir isotherm model for multicomponent adsorption equilibria which takes into account the vacant site as part of the adsorbed phase, this study presents the thermodynamic condition for adsorption azeotropes as derived from the generalized Langmuir isotherm to be the equality of the ratios of adsorbed phase activity coefficient γi and adsorption equilibrium constant Kio for the two adsorbates in the binary 1-2 adsorption system, i.e., γ1/K1o = γ2/K2o. This adsorption azeotropic condition is analogous to the vapor-liquid equilibrium azeotropic condition, i.e., the equality of the products of liquid phase activity coefficient and saturation vapor pressure Pisat for the two components, i.e., γ1P1sat = γ2P2sat. We validated the thermodynamic condition for adsorption azeotropes with 14 azeotrope-forming adsorption systems in this study. Furthermore, we investigated the effects of the pressure, temperature, and adsorbed phase nonideality on azeotrope formation.


Intracellular pH in the toad Bufo marinus following hypercapnia.
We investigated the effects of hypercapnia on intracellular acid-base regulation in brain and liver of the toad Bufo marinus L. After 1 h at 5% CO2, arterial PCO2 increased significantly, from 1.6 +/- 0.04 to 5.7 +/- 0.23 kPa, while brain and liver intracellular pH (pHi) decreased significantly. Reductions in pHi of both tissues were partially compensated by increased levels of bicarbonate. Surprisingly, however, compensation was lower than expected in brain and higher than expected in liver. We suggest that compensation in brain may be limited by secondary effects of bicarbonate loading in this tissue.


Cerebrospinal Fluid Findings in Patients With Autoimmune Encephalitis-A Systematic Analysis.
Autoimmune encephalitides (AIE) comprise a group of inflammatory diseases of the central nervous system (CNS), which can be further characterized by the presence of different antineuronal antibodies. Recently, a clinical approach for diagnostic criteria for the suspected diagnosis of AIE as well as definitive AIE were proposed. These are intended to guide physicians when to order the antineuronal antibody testing and/or facilitate early diagnosis even prior to the availability of the specific disease-confirming test results to facilitate prompt treatment. These diagnostic criteria also include the results of basic cerebrospinal fluid (CSF) analysis. However, the different antibody-defined AIE subtypes might be highly distinct with regard to their immune pathophysiology, e.g., the pre-dominance of specific IgG subclasses, IgG1, or IgG4, or frequency of paraneoplastic compared to idiopathic origin. Thus, it is conceivable that the results of basic CSF analysis might also be very different. However, this has not been explored systematically. Here, we systematically reviewed the literature about the 10 most important AIE subtypes, AIE with antibodies against NMDA, AMPA, glycine, GABAA, and GABAB receptors as well as DPPX, CASPR2, LGI1, IgLON5, or glutamate decarboxylase (GAD), with respect to the reported basic CSF findings comprising CSF leukocyte count, total protein, and the presence of oligoclonal bands (OCB) restricted to the CSF as a sensitive measure for intrathecal IgG synthesis. Our results indicate that these basic CSF findings are profoundly different among the 10 different AIE subtypes. Whereas, AIEs with antibodies against NMDA, GABAB, and AMPA receptors as well as DPPX show rather frequent inflammatory CSF changes, in AIEs with either CASPR2, LGI1, GABAA, or glycine receptor antibodies CSF findings were mostly normal. Two subtypes, AIEs defined by either GAD, or IgLON5 antibodies, did not fit into this general pattern. In AIE with GAD antibodies, positive OCBs in the absence of other changes were typical, while the CSF in IgLON5 antibody-positive AIE was characterized by elevated protein.


Primary cerebral neuroblastoma: a case treated with adjuvant chemotherapy and radiotherapy.
Neuroblastoma is the most common extracranial solid tumor of childhood. A wide variety of tumor locations and clinical presentations have been described. However, neuroblastoma is rarely located in the central nervous system, except in the case of esthesioneuroblastoma. We report a child with primary central nervous system (CNS) neuroblastoma who admitted to the hospital in coma. The tumor could be partially removed in our patient. After surgery, we treated the patient successfully with combined radiotherapy and eight courses of cisplatin-based chemotherapy. Our patient was followed-up for 34 months without any evidence of relapse.


Phase I trial of sequential topotecan followed by etoposide in adults with myeloid leukemia: a National Cancer Institute of Canada Clinical Trials Group Study.
Prolonged exposure to a topoisomerase I inhibitor may increase expression of topoisomerase II, making cells more susceptible inhibitors of that enzyme. This study was undertaken to establish the maximum tolerated dose (MTD) of a topotecan/topoisomerase II inhibitor sequential combination that may be active in acute leukemia, and to evaluate the effects of in vivo exposure to topotecan on topoisomerase II levels in leukemic blast cells as measured by image cytometry. Patients who were eligible for this phase I study had relapsed or refractory acute myeloid leukemia (< or = 2 prior regimens) or CML blast crisis (0 or 1 prior regimen). Topotecan was given as a 5 day continuous i.v. infusion and was to be escalated through three levels (1.5, 1.75 and 2.0 mg/m2 day), followed by etoposide at two dose levels (100 and 150 mg/m2) i.v. bolus days 6, 7 and 8. Topoisomerase IIalpha levels in leukemic blasts from bone marrow were measured by image cytometry prior to starting treatment, on day 5 of topotecan infusion and on day 28; and daily during topotecan in peripheral blood blasts. Dose-limiting toxicity was seen in two of six patients at the first dose level (topotecan 1.5 mg/m2/day, etoposide 100 mg/m2/day; > or = grade 3 mucositis in both cases). This cohort was expanded to 10 patients; no further non-hematologic dose-limiting toxicity was observed, but given the extent of toxicity seen, further dose escalation was judged not to be feasible. Topo IIalpha levels increased in peripheral blood blasts during the first 72 h of topotecan infusion and returned to near baseline by day 5, whereas levels appeared to decrease in bone marrow blasts by day 5 compared to pretreatment. One complete hematologic and cytogenetic remission in a patient with CML blast crisis was observed in the 10 patients evaluable for response. The sequential administration of topotecan 1.5 mg/m2/day continuous infusion for 5 days followed by etoposide 100 mg/m2/day x 3 is the recommended phase II dose for this schedule. Topotecan increases topo IIalpha expression in vivo in leukemia cells, but levels of the enzyme are cell cycle dependent. Pharmacodynamic evaluation of the sequential or combination administration of novel antileukemic agents may help improve treatment strategies in acute leukemia.


Generation of targeted nonpolar gene insertions and operon fusions in Pasteurella haemolytica and creation of a strain that produces and secretes inactive leukotoxin.
An efficient method for targeted gene inactivation and generation of chromosomal gene fusions in Pasteurella haemolytica has been devised and used to create an lktC::cat operon fusion by allelic exchange at the leukotoxin gene cluster (lktCABD). A copy of the lktC gene was insertionally inactivated by using a nonpolar, promoterless cat cassette and then delivered into P. haemolytica on a shuttle vector. Plasmid incompatibility was used to detect clones where double recombination events had occurred at the chromosomal locus. The insertion in lktC did not affect expression of the downstream genes, and the mutant strain secreted an antigenic proleukotoxin that was neither leukotoxic nor hemolytic. Expression of the lktC gene in trans restored the wild-type phenotype, confirming that LktC is required for activation of the proleukotoxin to the mature leukotoxin. Construction of the lktC::cat operon fusion allowed us to quantitate leukotoxin promoter activity in P. haemolytica and to demonstrate that transcription was maximal during early logarithmic growth phase but was reduced following entry into late logarithmic phase. This allelic exchange system should be useful for future genetic studies in P. haemolytica and could potentially be applied to other members of Haemophilus-Actinobacillus-Pasteurella family, where genetic manipulation is limited.


Mechanical response and conformational amplification in α-helical coiled coils.
α-Helical coiled coils (CCs) are ubiquitous tertiary structural domains that are often found in mechanoproteins. CCs have mechanical rigidity and are often involved in force transmission between protein domains. Although crystal structures of CCs are available, information about their conformational flexibility is limited. The role of hydrophobic interactions in determining the CC conformation is not clear. In this work we examined the mechanical responses of typical CCs and constructed a coarse-grained mechanical model to describe the conformation of the protein. The model treats α-helices as elastic rods. Hydrophobic bonds arranged in a repeated pattern determine the CC structure. The model is compared with molecular-dynamics simulations of CCs under force. We also estimate the effective bending and twisting persistence length of the CC. The model allows us to examine unconventional responses of the CC, including significant conformational amplification upon binding of a small molecule. We find that the CC does not behave as a simple elastic rod and shows complex nonlinear responses. These results are significant for understanding the role of CC structures in chemoreceptors, motor proteins, and mechanotransduction in general.


The relationship between sun protection policies and practices in schools with primary-age students: the role of school demographics, policy comprehensiveness and SunSmart membership.
Schools can implement evidence-based sun protection policies that guide practices to help protect children from harmful sun exposure. This national study assessed the relationship between the existence and comprehensiveness of written policies and the comprehensiveness of sun protection practices. The impact of school demographics on the strength of the relationship was also examined, as was the possibility that 'SunSmart' membership would have an additional impact on practices, beyond having any formal policy. In 2011-12, staff members of 1573 schools catering to primary-age students completed a self-administered survey about sun protection policies and practices (response rate of 57%). Results showed that schools with a written policy had more comprehensive practices than schools without a written policy. The relationship between having a written policy and sun protection practices was stronger for remote schools compared with metropolitan and regional schools, and for schools catering to both primary and secondary students compared with primary students only. In addition, policy comprehensiveness was associated with practice comprehensiveness, and SunSmart membership was indirectly related to practice comprehensiveness via policy comprehensiveness. These results indicate that written policies relate to practice comprehensiveness, but the strength of the association can vary according to the characteristics of the organization.


An investigation into the effect of the structure of bile salt aggregates on the binding interactions and ESIHT dynamics of curcumin: a photophysical approach to probe bile salt aggregates as a potential drug carrier.
This work demonstrates the utilization of bile salt aggregates as a potential biological host system for studying the binding interactions and dynamics of the poorly-water-soluble drug curcumin by means of photophysical techniques. We found that the level of degradation of curcumin is greatly suppressed upon encapsulation into the nanocavities of three different bile salt aggregates. However, NaTC aggregates are more effective to suppress the level of degradation of curcumin than NaCh and NaDC aggregates. We also report the modulation of the photophysical and dynamical properties of curcumin into the nanocavities of bile salt aggregates using steady-state and time-resolved fluorescence spectroscopy. The reduced level of interaction of curcumin with water upon incorporation into the different binding sites of bile salt aggregates results in an enhanced fluorescence intensity along with the blue shift in the emission maxima of curcumin. However, the observation of higher fluorescence quantum yield as well as longer fluorescence lifetime in NaTC aggregates compared to that in NaCh and NaDC aggregates clearly indicates a more effective decrease in the excited-state intramolecular hydrogen atom transfer (ESIHT) mediated nonradiative deactivation of curcumin by the interaction with the anionic headgroup of NaTC. The binding and location of curcumin into the bile salt aggregates has been further confirmed from the steady-state fluorescence anisotropy measurements. In addition, we have shown the effect of addition of salt on the photophysical properties of curcumin in the confined environments of bile salt aggregates. Our results indicate that on addition of salt the time scale of ESIHT process of curcumin in bile salt aggregates is markedly increased.


In vivo Ca2+ dynamics in a cricket auditory neuron: an example of chemical computation.
Fura-2 calcium imaging in the cricket omega neuron revealed increased intracellular free calcium ion concentration in response to simulated cricket calling songs and other sound stimuli. The time course of the increase and decrease in intracellular calcium coincided with the time course of forward masking, a time-dependent modulation of auditory sensitivity. The buffering of calcium transients with high concentrations of a kinetically fast calcium buffer eliminated the post-stimulus hyperpolarization associated with forward masking, whereas the uncaging of calcium inside the neuron produced a hyperpolarization. The results suggest that sound-stimulated intracellular calcium accumulation acts by means of a calcium-activated hyperpolarizing current to produce forward masking. These findings underscore the importance of chemical dynamics in neural computation by demonstrating a behaviorally relevant role of calcium dynamics in vivo.


Zuclopenthixol decanoate in schizophrenia: serum levels and clinical state.
Serum concentrations of zuclopenthixol were determined in a group of 20 patients treated with a depot preparation, zuclopenthixol decanoate in Viscoleo. Clinical assessments according to a Clinical Global Impression (CGI) scale, Comprehensive Psychological Rating Scale (CPRS), 16-item subscale for schizophrenia, and the UKU side effect scale were performed on 3 consecutive days of injection. The serum concentrations showed a limited individual variation and a high and significant correlation between dose and serum concentration. One patient had a particularly high serum concentration of zuclopenthixol. This patient also had an elevated concentration of the N-dealkyl metabolite, but a low concentration of the sulphoxide. For serum concentrations versus clinical state and side effects some significant correlations were found. All correlations were positive, which means that the higher the serum concentration the poorer the clinical state of the patient. We think that this probably reflects a common clinical pattern of increasing the dose, when the antipsychotic response is unsatisfactory. The study also showed that for moderately ill patients, who were given the optimum dose of drug, the subgroup of patients not experiencing side effects had significantly lower serum concentrations than the subgroup with side effects.


Effects of an animated educational video on knowledge of cell-free DNA screening among Thai pregnant women: a randomized control trial.
In developing countries, pregnant women have insufficient knowledge about cell-free DNA screening. Reports from developed countries have found that various tools in prenatal genetic counseling can improve the knowledge of pregnant women who undergo cell-free DNA screening. Data are limited from developing countries where women have different baseline socio-educational backgrounds. The objective of this study was to compare the effects of an animated educational video combined with traditional counseling versus traditional counseling alone in changing pregnant women's knowledge of cell-free DNA screening. This study was a randomized control trial at an antenatal clinic. Eligible subjects who were Thai pregnant women, were randomized to either view or not view the 4-minute animated educational video explaining cell-free DNA screening. Both groups received traditional counseling. The women were asked to complete a Thai questionnaire assessing knowledge of the screening before and after intervention. The questionnaire consisted of three sections: demographic data of the research participants and their existing awareness about cell-free DNA testing; performance and limitations of cell-free DNA screening; and participants' attitudes toward the positive screening. Primary outcome was the change in knowledge scores. Secondary outcomes were attitudes toward positive screening test, levels of satisfaction with counseling, and screening acceptance rates. Data from 83 women in the video group and 82 in the non-video group were analyzed. The knowledge score (range 0-18) change after counseling was significantly higher in the video group than the non-video group (+ 7.1 ± 3.3 vs + 4.2 ± 2.5; p = 0.03). There were no significant differences in attitudes toward positive screening test (p = 0.83), levels of satisfaction (p = 0.24), or screening acceptance rates (p = 0.15) between the groups. Adding the video to traditional counseling was better than traditional counseling alone in improving pregnant women's knowledge about cell-free DNA screening. The study was retrospectively registered with the Thai Clinical Trials Registry (TCTR20210917001, 17/09/2021).


[Relation between cervical mucus sperm toxicity and pregnancy in cows].
Cervical secretion was collected from 99 dairy cows of the Black Pied Lowland breed at an age of 24 to 109 months with a minimum performance of 4000 kg of milk annually in spontaneous oestrus 42 days post partum at the minimum. It was demonstrated by the radioimmunological determination of progesterone in skim milk that from all the breeding cows (n = 99) the oestral secretion was collected during an optimum insemination time (x = 0.19 ng.ml-1 progesterone). Of the statistical set of 99 cows, 55 (i.e. 56.57%) got in calf and 43 (i.e. 43.43%) remained barren. As found, the conception results were not influenced by the rank of the insemination after parturition and the conception capacity of the cows was not influenced by the length of time from parturition to the respective inseminations. It was demonstrated in our investigation that the conception capacity of the cows increases as the time of sperm survival prolongs in the cervical secretion in vitro. The highest conception capacity (80%) was recorded in the cows whose cervical secretion was free of spermiotoxic action. In such cases the motility of the spermatozoa in the oestral mucus was maintained at a temperature of 38 degrees C for longer than 300 minutes. On the other hand, it was demonstrated that the conception capacity of the cows declines with shortening time of sperm survival in the cervical mucus.


[Cosmetic dentistry and adhesive prosthodontics].
A better chemical bonding to metals offers the opportunity to bind composites onto metal and metals onto tooth structures. This may lead to an increase of adhesive prosthetic and cosmetic treatments. In this article some applications of this modern treatment method are described, anterior and posterior, in solitary elements as well as multiple bridge constructions.


Results of a 2-year retrospective cohort study of newly prescribed triptan users in European nationwide practice databases.
This study was conducted to characterize prescription refill patterns for triptans among European patients with new prescriptions of triptans. Persistency with prescriptions of triptan monotherapy for migraine headache among newly prescribed users in European primary-care practices has not been well described. Using electronic medical databases in the UK (N = 3618), France (N = 2051) and Germany (N = 954), we conducted a retrospective cohort analysis to identify refill patterns over 2 years among migraineurs receiving new prescriptions of triptan monotherapy in 2006. Of all patients, >33% of migraineurs with new triptan prescriptions received ≥1 refill of their index triptan prescriptions (UK, 44.3%; France, 34.2%; Germany, 37.7%). More than 50% never received index-triptan refill prescriptions (UK, 55.7%; France, 65.8%; Germany, 63.3%). Small proportions of patients (<7.0%) switched to alternative triptans, and even fewer switched to different prescription-medication classes (UK and Germany, 2.3%; France, 4.0%). More than 48% of patients received no further prescriptions for migraine after index prescriptions (UK, 48.5%; France, 54.9%; Germany, 54.7%). After the second year, >83.0% of patients in each country had no further prescriptions for migraine medications, <14.0% remained persistent with index prescriptions, <4.0% switched to other triptans, and <3.0% switched to alternative medication classes. In migraine patients who received new prescriptions of triptan monotherapy from their primary-care physicians, poor triptan prescription refill frequency was observed in Europe. Although consistent with potential clinical challenges in migraine management, our findings should be interpreted with caution given certain inherent limitations associated with the database study design. Further research is warranted to confirm our findings and to identify reasons for, or predictors of, triptan discontinuation.


Increased Skeletal Muscle Capillarization Independently Enhances Insulin Sensitivity in Older Adults After Exercise Training and Detraining.
Intramuscular signaling and glucose transport mechanisms contribute to improvements in insulin sensitivity after aerobic exercise training. This study tested the hypothesis that increases in skeletal muscle capillary density (CD) also contribute to exercise-induced improvements in whole-body insulin sensitivity (insulin-stimulated glucose uptake per unit plasma insulin [M/I]) independent of other mechanisms. The study design included a 6-month aerobic exercise training period followed by a 2-week detraining period to eliminate short-term effects of exercise on intramuscular signaling and glucose transport. Before and after exercise training and detraining, 12 previously sedentary older (65 ± 3 years) men and women underwent research tests, including hyperinsulinemic-euglycemic clamps and vastus lateralis biopsies. Exercise training increased Vo2max (2.2 ± 0.2 vs. 2.5 ± 0.2 L/min), CD (313 ± 13 vs. 349 ± 18 capillaries/mm(2)), and M/I (0.041 ± 0.005 vs. 0.051 ± 0.007 μmol/kg fat-free mass/min) (P < 0.05 for all). Exercise training also increased the insulin activation of glycogen synthase by 60%, GLUT4 expression by 16%, and 5' AMPK-α1 expression by 21%, but these reverted to baseline levels after detraining. Conversely, CD and M/I remained 15% and 18% higher after detraining, respectively (P < 0.05), and the changes in M/I (detraining minus baseline) correlated directly with changes in CD in regression analysis (partial r = 0.70; P = 0.02). These results suggest that an increase in CD is one mechanism contributing to sustained improvements in glucose metabolism after aerobic exercise training.


Modifiable Risk Factors and Trends in Changes in Glucose Regulation during the First Three Years Postdelivery: The St Carlos Gestational Diabetes Mellitus Prevention Cohort.
Evaluation of the influence of potential risk factors (RFs) on glycemic changes at 3 years postpartum. The glycemic status of 1400 women, in absence of a new pregnancy, was evaluated at 3 months (3 m) and 3 years (3 y) postpartum, after participation in the St. Carlos Gestational Study (2228 normoglycemic pregnant women followed from before gestational week 12 to delivery, from 2015-2017). Abnormal glucose regulation (AGR) was defined as fasting serum glucose ≥ 100 mg/dL and/or HbA1c ≥ 5.7% and/or 2 h 75 g OGTT glucose ≥ 140 mg/dL. In total, 12 modifiable and 3 unmodifiable RFs were analyzed. 3 m postpartum, 110/1400 (7.9%) women had AGR; 3 y postpartum, 137 (9.8%) women exhibited AGR (110 with 3 m normal glucose tolerance [NGT]); 1263 (90.2%) had NGT (83 with 3 m AGR). More women with gestational diabetes mellitus (GDM) progressed to AGR at 3 y (OR: 1.60 [1.33-1.92]) than women without GDM. Yet, most women with 3 m and/or 3 y AGR had no GDM history. Having ≥2 unmodifiable RFs was associated with increased risk for progression to AGR (OR: 1.90 [1.28-2.83]) at 3 y postpartum. Having >5/12 modifiable RFs was associated with increased progression from NGT to AGR (OR: 1.40 [1.00-2.09]) and AGR persistence (OR: 2.57 [1.05-6.31]). Pregestational BMI ≥ 25 kg/m2 (OR: 0.59 [0.41-0.85]), postdelivery weight gain (OR: 0.53 [0.29-0.94]), and waist circumference > 89.5 cm (OR: 0.54 [0.36-0.79]) reduced the likelihood of NGT persisting at 3 y. 3-month and/or 3-year postpartum AGR can be detected if sought in women with no prior GDM. Modifiable and unmodifiable RF predictors of AGR at 3 y postpartum were identified. Universal screening for glycemic alterations should be considered in all women following delivery, regardless of prior GDM. These findings could be useful to design personalized strategies in women with risk factors for 3 y AGR.


Temporal bone histopathological findings in trisomy 13 syndrome.
This study reports the histopathological findings of 14 temporal bones from infants with trisomy 13 syndrome. The most primitive anomalies in the structures of the inner and middle ears in the present series are those of the semicircular canals, particularly of the horizontal canals: flattened horizontal canal cristae, absence or opening of the utricular endolymphatic valve, small facial nerve, and obtuse angle of the geniculate area of the facial nerve. Each ear demonstrated more than one of those anomalies. The anomalies present features similar to those found in the structures of the normal six to ten-week fetus. Many other mild anomalies observed appear to demonstrate features similar to those seen in the same structures in later fetal life. Reviewing these findings, most of the anomalies that were found in the inner and middle ears appear to be the result of poor development of the structures for reasons which are now unclear. In addition, middle ear infection was found in all cases.


Comparative Serum Challenges Show Divergent Patterns of Gene Expression and Open Chromatin in Human and Chimpanzee.
Humans experience higher rates of age-associated diseases than our closest living evolutionary relatives, chimpanzees. Environmental factors can explain many of these increases in disease risk, but species-specific genetic changes can also play a role. Alleles that confer increased disease susceptibility later in life can persist in a population in the absence of selective pressure if those changes confer positive adaptation early in life. One age-associated disease that disproportionately affects humans compared with chimpanzees is epithelial cancer. Here, we explored genetic differences between humans and chimpanzees in a well-defined experimental assay that mimics gene expression changes that happen during cancer progression: A fibroblast serum challenge. We used this assay with fibroblasts isolated from humans and chimpanzees to explore species-specific differences in gene expression and chromatin state with RNA-Seq and DNase-Seq. Our data reveal that human fibroblasts increase expression of genes associated with wound healing and cancer pathways; in contrast, chimpanzee gene expression changes are not concentrated around particular functional categories. Chromatin accessibility dramatically increases in human fibroblasts, yet decreases in chimpanzee cells during the serum response. Many regions of opening and closing chromatin are in close proximity to genes encoding transcription factors or genes involved in wound healing processes, further supporting the link between changes in activity of regulatory elements and changes in gene expression. Together, these expression and open chromatin data show that humans and chimpanzees have dramatically different responses to the same physiological stressor, and how a core physiological process can evolve quickly over relatively short evolutionary time scales.


Evaluation of predicted network modules in yeast metabolism using NMR-based metabolite profiling.
Genome-scale metabolic models promise important insights into cell function. However, the definition of pathways and functional network modules within these models, and in the biochemical literature in general, is often based on intuitive reasoning. Although mathematical methods have been proposed to identify modules, which are defined as groups of reactions with correlated fluxes, there is a need for experimental verification. We show here that multivariate statistical analysis of the NMR-derived intra- and extracellular metabolite profiles of single-gene deletion mutants in specific metabolic pathways in the yeast Saccharomyces cerevisiae identified outliers whose profiles were markedly different from those of the other mutants in their respective pathways. Application of flux coupling analysis to a metabolic model of this yeast showed that the deleted gene in an outlying mutant encoded an enzyme that was not part of the same functional network module as the other enzymes in the pathway. We suggest that metabolomic methods such as this, which do not require any knowledge of how a gene deletion might perturb the metabolic network, provide an empirical method for validating and ultimately refining the predicted network structure.


Role of quinones in the branch of the Escherichia coli respiratory chain that terminates in cytochrome o.
The role of quinones in the cytochrome o branch of the Escherichia coli respiratory chain was investigated by using mutant strains lacking the cytochrome d terminal oxidase complex. The only cytochromes present were cytochrome b556 and the cytochrome o complex, consisting of cytochrome b555-b562. Mutant strains missing ubiquinone, menaquinone, or both were constructed in the cytochrome d-minus (cyd) background. The steady-state levels of cytochrome b reduction were examined and compared in these strains to assess the effects of the quinone deficiencies. The data clearly show that a ubiquinone deficiency results in a lower level of cytochrome b reduction in the steady state. The data are consistent with a simple model in which ubiquinone is placed on the dehydrogenase side of all the cytochromes in this branch of the respiratory chain. There is no evidence from these experiments for a role of quinones in the respiratory chain at any site besides this one.


Determination of Pesticides in Soybean Seeds Incorrectly Discarded Near a Spring of the Paranaíba River, GO-Brazil.
The objective of this research was to evaluate the contamination of pesticides of carbamate and benzimidazole classes in soil, water and soybean seeds, incorrectly discarded near a spring in the state of Goiás-Brazil. The Solid-Liquid and Liquid-Liquid Extraction with Low Temperature Partition (SLE/LTP or LLE/LTP) methods were used for pesticide extraction and the analyses were performed by HPLC-UV. A high resolution mass spectrometer was used to confirm the identity of the compounds present in the seeds. The results showed that the soybeans were treated with the pesticide carbendazim and the dosage was three times higher than established by Brazilian legislation. In the soil and water analyzed there was no presence of the pesticides researched, nor of the carbendazim detected in the seeds. Since this was an environmental crime due to incorrect disposal, it is not known how long ago it occurred. Thus, depending on how long the seeds had been exposed there, the pesticide could have leached out, and caused contamination in the spring and soil. Thus, we conclude that the incorrect disposal of seeds treated with pesticides can be a risk to the permanence of life on the site.


Circadian regulation of hippocampal long-term potentiation.
The goal of this study is to investigate the possible circadian regulation of hippocampal excitability and long-term potentiation (LTP) measured by stimulating the Schaffer collaterals (SC) and recording the field excitatory postsynaptic potential (fEPSP) from the CA1 dendritic layer or the population spike (PS) from the soma in brain slices of C3H and C57 mice. These 2 strains of mice were of interest because the C3H mice secrete melatonin rhythmically while the C57 mice do not. The authors found that the magnitude of the enhancement of the PS was significantly greater in LTP recorded from night slices compared to day slices of both C3H and C57 mice. They also found significant diurnal variation in the decay of LTP measured with fEPSPs, with the decay slower during the night in both strains of mice. There was evidence for a diurnal rhythm in the input/output function of pyramidal neurons measured at the soma in C57 but not C3H mice. Furthermore, LTP in the PS, measured in slices prepared during the day but recorded during the night, had a profile remarkably similar to the night group. Finally, PS recordings were carried out in slices from C3H mice maintained in constant darkness prior to experimentation. Again, the authors found that the magnitude of the enhancement of the PS was significantly greater in LTP recorded from subjective night slices compared to subjective day slices. These results provide the 1st evidence that an endogenous circadian oscillator modulates synaptic plasticity in the hippocampus.