The left intraparietal sulcus adapts to symbolic number in both the visual and auditory modalities: Evidence from fMRI.
A growing body of evidence from functional Magnetic Resonance Imaging adaptation (fMRIa) has implicated the left intraparietal sulcus (IPS) as a crucial brain region representing the semantic of number symbols. However, it is currently unknown to what extent the left IPS brain activity can be generalized across modalities (e.g., Arabic digits and spoken number words) and how robust and reproducible numerical adaptation effects are. In two separate fMRIa experiments we habituated the brain response of 20 native English-speaking (Experiment 1) and 34 native German-speaking (Experiment 2) adults to Arabic digits or spoken number words. Consistent with previous findings, experiment 1 revealed numerical ratio dependent adaptation to Arabic numerals in the left IPS using both conventional and cortex-based alignment techniques. Experiment 2 revealed numerical ratio dependent signal recovery in the left IPS following adaptation to both Arabic numerals and spoken number words using both conventional and cortex-based alignment techniques. Together, these findings suggest that the left IPS is involved in symbolic number processing across modalities.


[Mitral valve replacement in an infant with disseminated lupus erythematosus and Libman-Sacks endocarditis].
The authors report a case of Libman-Sacks endocarditis of the posterior mitral leaflet in a child treated for disseminated lupus erythematosus for 8 years. Mitral valve replacement was performed. Libman-Sacks vegetations are hardly ever recognised in vivo and have never been previously reported in a child. The incidence of this cardiac complication of lupus erythematosus is probably higher than indicated in the literature. Attentive cardiac examination and systematic echocardiography should be part of routine follow-up of patients with disseminated lupus erythematosus.


Molecular modeling of metal complexation by a fluoroquinolone antibiotic.
An understanding of the factors controlling the chemodynamics of fluoroquinolone antibiotics in different environmental matrices is a necessary prerequisite to the assessment of their potential impact on nontarget organisms in soils and receiving waters. Of particular interest are the complexes formed between fluoroquinolones and metal cations, which are believed to be important in the mechanism of sequestration of the antibiotic by minerals and natural organic matter. The structures of these complexes have not been fully resolved by conventional spectroscopy; therefore, molecular simulations may provide useful complementary insights. We present results from apparently the first molecular dynamics simulations of a widely used fluoroquinolone antibiotic, ciprofloxacin (Cipro), in aqueous complexes with five metal cations typically found in soils and surface waters: Ca2+, Mg2+, Fe2+, Na+, and K+. The interatomic potential functions employed in the simulations were validated by comparison with available structural data for solid-phase Cipro-hexahydrate and for the metal cations in aqueous solution. Although no comprehensive structural data on the aqueous complexes appear to be available, properties of the metal complexes predicted by our simulations agree with available data for solid-phase metal-Cipro complexes. Our results indicate that the ionic potential of the metal cation controls the stability of the complex formed and that the hydration number of the metal cation in aqueous solution determines its coordination number with O atoms in the metal-Cipro complex. In respect to environmental chemodynamics, our results imply that Cipro will form two configurations of bidendate chelates with metal centers on exposed surfaces of mineral oxides, water-bridged surface complexes with exchangeable cations in clay mineral interlayers, and cation-bridged complexes with functional groups in natural organic matter.


Aetiology of headache in clinical ophthalmic practice at a tertiary care hospital of Karachi.
To determine the aetiology of headache in patients seen for an ocular examination. This cross-sectional, descriptive study was conducted at Ophthalmology Department of Karachi Medical and Dental College, Abbasi Shaheed Hospital, Karachi, from January to December 2014, and comprised patients with headache. Patients were registered through non-probability consecutive sampling technique. A predesigned proforma was used to collect data. Complete ocular examination and investigations were conducted along with neuro-ophthalmological examination. Data analysis was done using SPSS 16. Of the 379 patients, 225(59.4%) were female and 154(40.6%) were male. The mean age was 35.12±16.387 years (range: 6-75 years). Conditions associated with headache were divided as ocular, non-ocular and combined pathologies. Among ocular causes, asthenopia was a major entity as 62(16.36%) asthenopic patients presented with refractive errors. These were followed by presbyopics who presented with complaint of headache 56(14.78%). The number of computer users with similar complaint was 18(4.76%). Among other ocular causes of headache, the number of patients with corneal ulcers was 22(5.80%), glaucoma 15(3.96%) and endophthalmitis 4(1.06%). Among the non-ocular causes were hypertension 59(15.57%), sinusitis 41(10.82%) and migraine 47(12.4%). The majority of patients had associate ocular causes such as refractive errors and anterior segment pathologies.


Current management of congenital diaphragmatic hernia.
The treatment of congenital diaphragmatic hernia (CDH) still represents a challenge, even for the specialised multidisciplinary teams in centres that provide treatment for CDH. Despite significant progress in the fields of pathophysiology, prenatal diagnosis, surgical techniques and intensive care, CDH is a disease still burdened with a high mortality. Due to the paucity of randomised studies, there are no standard guidelines for treatment. The present review looks at existing diagnostic and therapeutic principles based on the available literature.


Is the multicolored Asian ladybeetle, Harmonia axyridis, the most abundant natural enemy to aphids in agroecosystems?
The multicolored Asian ladybeetle, Harmonia axyridis Pallas (Coleoptera: Coccinellidae), was introduced into Western Europe in the late 1990s. Since the late 2000s, this species has been commonly considered one of the most abundant aphid predators in most Western European countries. In spite of the large amount of research on H. axyridis, information concerning its relative abundance in agroecosystems is lacking. This study aims to evaluate the abundance of H. axyridis within the aphidophage community in four crops situated in southern Belgium: wheat, Triticum aestivum L. (Poales: Poaceae), corn, Zea mays, potato, Solanum tuberosum (Solanales: Solanaceae), and broad bean Vicia faba (Fabales: Fabaceae). In order to assess the species diversity, the collected data were analyzed by considering (1) the species richness and (2) the evenness according to the Shannon diversity index. Eleven aphidophages were observed in every inventoried agroecosystem, including five abundant species: three coccinellids, the seven-spotted ladybug, Coccinella septempunctata L. (Coleoptera: Coccinellidae), the 14-spotted Ladybird, Propylea quatuordecimpunctata, and H. axyridis; one hoverfly, the marmalade hoverfly, Episyrphus balteatus De Geer (Diptera: Syrphidae); and one lacewing, the common green lacewing, Chrysoperla carnea Stephens sensu lato (= s.l.) (Neuroptera: Chrysopidae). Harmonia axyridis has been observed to thrive, breed, and reproduce on the four studied crops. Harmonia axyridis is the most abundant predator of aphids in corn followed by C. septempunctata, which is the main aphid predator observed in the three other inventoried crops. In wheat and potato fields, H. axyridis occurs in low numbers compared to other aphidophage. These observations suggest that H. axyridis could be considered an invasive species of agrosystems, and that potato and wheat may intermittently act as refuges for other aphidophages vulnerable to intraguild predation by this invader. Harmonia axyridis is not the most abundant aphid predator in the main Belgian crops.


Effect of methylation on the stability and solvation free energy of amylose and cellulose fragments: a molecular dynamics study.
Molecular dynamics (MD) simulations were used to study the stability and solvation of amylose and cellulose fragments. The recently developed gromos carbohydrate force field was further tested by simulating maltose, cellobiose, and maltoheptaose. The MD simulations reproduced fairly well the favorable conformations of disaccharides defined by the torsional angles related with the glycosidic bond and the radius gyration of maltoheptaose. The effects of methylation at different hydroxyl groups on the stability of amylose and cellulose fragments were investigated. The methylations of O-2 and O-3 reduce the stability of a single helix more than methylation at O-6, while the latter reduces the stability of a double helix more. Solvation free-energy differences between the unsubstituted amylose and cellulose fragments and the methylated species were studied using the single-step perturbation method. It was found that methylation at O-2 has the biggest effect, in agreement with experiment.


Effect of denosumab on bone mineral density and biochemical markers of bone turnover: 8-year results of a phase 2 clinical trial.
In a phase 2 study, continued denosumab treatment for up to 8 years was associated with continued gains in bone mineral density and persistent reductions in bone turnover markers. Denosumab treatment was well tolerated throughout the 8-year study. The purpose of this study is to present the effects of 8 years of continued denosumab treatment on bone mineral density (BMD) and bone turnover markers (BTM) from a phase 2 study. In the 4-year parent study, postmenopausal women with low BMD were randomized to receive placebo, alendronate, or denosumab. After 2 years, subjects were reallocated to continue, discontinue, or discontinue and reinitiate denosumab; discontinue alendronate; or maintain placebo for two more years. The parent study was then extended for 4 years where all subjects received denosumab. Of the 262 subjects who completed the parent study, 200 enrolled in the extension, and of these, 138 completed the extension. For the subjects who received 8 years of continued denosumab treatment, BMD at the lumbar spine (N = 88) and total hip (N = 87) increased by 16.5 and 6.8 %, respectively, compared with their parent study baseline, and by 5.7 and 1.8 %, respectively, compared with their extension study baseline. For the 12 subjects in the original placebo group, 4 years of denosumab resulted in BMD gains comparable with those observed during the 4 years of denosumab in the parent study. Reductions in BTM were sustained over the course of continued denosumab treatment. Reductions also were observed when the placebo group transitioned to denosumab. Adverse event profile was consistent with previous reports and an aging cohort. Continued denosumab treatment for 8 years was associated with progressive gains in BMD, persistent reductions in BTM, and was well tolerated.


Rhizobium sp. strain NGR234 NodZ protein is a fucosyltransferase.
Rhizobium sp. strain NGR234 produces a large family of lipochitooligosaccharide Nod factors carrying specific substituents. Among them are 3-O- (or 4-O-) and 6-O-carbamoyl groups, an N-methyl group, and a 2-O-methylfucose residue which may bear either 3-O-sulfate or 4-O-acetyl substitutions. Investigations on the genetic control of host specificity revealed a number of loci which directly affect Nod factor structure. Here we show that insertion and frameshift mutations in the nodZ gene abolish fucosylation of Nod factors. In vitro assays using GDP-L-fucose as the fucose donor show that fucosyltransferase activity is associated with the nodZ gene product (NodZ). NodZ is located in the soluble protein fraction of NGR234 cells. Together with extra copies of the nodD1 gene, the nodZ gene and its associated nod box were introduced into ANU265, which is NGR234 cured of the symbiotic plasmid. Crude extracts of this transconjugant possess fucosyltransferase activity. Fusion of a His6 tag to the NodZ protein expressed in Escherichia coli yielded a protein able to fucosylate both nonfucosylated NodNGR factors and oligomers of chitin. NodZ is inactive on monomeric N-acetyl-D-glucosamine and on desulfated Rhizobium meliloti Nod factors. Kinetic analyses showed that the NodZ protein is more active on oligomers of chitin than on nonfucosylated NodNGR factors. Pentameric chitin is the preferred substrate. These data suggest that fucosylation occurs before acylation of the Nod factors.


Brain-derived neurotrophic factor/tropomyosin receptor kinase B signaling in spinal muscular atrophy and amyotrophic lateral sclerosis.
Tropomyosin receptor kinase B (TrkB) and its primary ligand brain-derived neurotrophic factor (BDNF) are expressed in the neuromuscular system, where they affect neuronal survival, differentiation, and functions. Changes in BDNF levels and full-length TrkB (TrkB-FL) signaling have been revealed in spinal muscular atrophy (SMA) and amyotrophic lateral sclerosis (ALS), two common forms of motor neuron diseases that are characterized by defective neuromuscular junctions in early disease stages and subsequently progressive muscle weakness. This review summarizes the current understanding of BDNF/TrkB-FL-related research in SMA and ALS, with an emphasis on their alterations in the neuromuscular system and possible BDNF/TrkB-FL-targeting therapeutic strategies. The limitations of current studies and future directions are also discussed, giving the hope of discovering novel and effective treatments.


Intracranial hemorrhage from giant aneurysm in pregnancy: A rare association.
We report a case of giant aneurysm causing subarachnoid hemorrhage in a pregnant female. A 25-year-old female presented with sudden onset of severe headache and vomiting with altered sensorium and right hemiplegia. On investigation, she had a giant supraclinoid segment internal carotid artery (ICA) aneurysm. She was planned for digital substraction angiography, but during the procedure she deteriorated neurologically and went into spontaneous labor. The baby was a male child with weight of 1.1 kg. She was taken up for surgery and aneurysm was clipped. We discuss the rare occurrence of intracranial hemorrhage in pregnancy due to a giant ICA aneurysm.


Suppression of cyclin D1 by hypoxia-inducible factor-1 via direct mechanism inhibits the proliferation and 5-fluorouracil-induced apoptosis of A549 cells.
Hypoxia-inducible factor (HIF) and cyclin D1 are both key mediators of cell growth and proliferation in normal and cancer cells. However, the interrelation between HIF and cyclin D1 remains unclear. In the present study, we observed the inverse correlation between cyclin D1 and HIF-1 in hypoxia condition. Overexpression of the dominant negative mutant of HIF-1alpha (DN-HIF) significantly enhanced cyclin D1 expression upon hypoxia or arsenite exposure, suggesting the negative regulation of cyclin D1 by HIF-1. Furthermore, we found that the impairment of HIF-1 increased cyclin D1 expression in A549 pulmonary cancer cells, which in turn promoted G1-S cell cycle transition and cell proliferation. Cyclin D1 expression was increased in s.c. xenograft of DN-HIF stably transfected A549 cells in nude mice compared with that of control cells. Chromatin immunoprecipitation assay revealed that HIF-1 was able to directly bind to the promoter region of cyclin D1, which indicates that the negative regulation of cyclin D1 by HIF-1 is through a direct mechanism. Inhibition of histone deacetylase (HDAC) by pretreatment of cells with trichostatin A or specific knockdown of HDAC7 by its shRNA antagonized the suppression of cyclin D1 by HIF-1, suggesting that HDAC7 is required for HIF-1-mediated cyclin D1 downregulation. Moreover, we found that 5-fluorouracil-triggered apoptosis of DN-HIF-transfected A549 cells was reduced by sicyclin D1 (cyclin D1-specific interference RNA) introduction, suggesting that clinical observation of HIF-1 overexpression-associated chemoresistance might be, at least partially, due to the negative regulation of cyclin D1.


Overview of chemoradiation clinical trials for locally advanced non-small cell lung cancer in Japan.
The standard of care for unresectable stage III non-small cell lung cancer (NSCLC) is combined-modality therapy with both chemotherapy and thoracic radiation therapy (TRT). A phase III trial by the West Japan Lung Cancer Group revealed that the combination of mitomycin, vindesine, and cisplatin (MVP) with concurrent TRT yielded a median survival time of 16.6 months and a 5-year survival rate of 16% in patients with unresectable stage III NSCLC. Although evidence indicates that concurrent chemotherapy and TRT (chemoradiation) increases survival to a moderately greater extent than sequential therapeutic approaches, the optimal strategies for such concurrent treatment remain to be defined, and differ between full-dose systemic and low-dose radio-enhancing protocols. Two phase III trials have been initiated in Japan to address these issues and they have recently reported preliminary data. Early results of the Okayama Lung Cancer Study Group (OLCSG) trial, comparing chemoradiation based on divided docetaxel and cisplatin chemotherapy with MVP-based chemoradiation, have been reported. The West Japan Oncology Group (WJOG) is comparing the efficacy and toxicity of TRT and concurrent chemotherapy with either carboplatin-paclitaxel or carboplatin-irinotecan, followed by full-dose consolidation chemotherapy, with the efficacy and toxicity of MVP-based chemoradiation. Several phase I/II studies to test the optimal use of new agents such as S-1 (an oral anticancer drug combining tegafur, 5-chloro-2, 4-dihydroxypyridine, and potassium oxonate) and gefitinib (an inhibitor of the tyrosine kinase activity of the epidermal growth factor receptor) are also ongoing. In addition, radiation dose intensification with three-dimensional planning approaches is currently under evaluation. A phase I clinical trial by WJOG to establish, prospectively, the maximum tolerated dose of three-dimensional hyperfractionated radiotherapy with concurrent weekly chemotherapy (carboplatin-paclitaxel) is thus currently under way. This overview of ongoing trials highlights new directions in the treatment of locally advanced NSCLC.


A novel test for gene-ancestry interactions in genome-wide association data.
Genome-wide association study (GWAS) data on a disease are increasingly available from multiple related populations. In this scenario, meta-analyses can improve power to detect homogeneous genetic associations, but if there exist ancestry-specific effects, via interactions on genetic background or with a causal effect that co-varies with genetic background, then these will typically be obscured. To address this issue, we have developed a robust statistical method for detecting susceptibility gene-ancestry interactions in multi-cohort GWAS based on closely-related populations. We use the leading principal components of the empirical genotype matrix to cluster individuals into "ancestry groups" and then look for evidence of heterogeneous genetic associations with disease or other trait across these clusters. Robustness is improved when there are multiple cohorts, as the signal from true gene-ancestry interactions can then be distinguished from gene-collection artefacts by comparing the observed interaction effect sizes in collection groups relative to ancestry groups. When applied to colorectal cancer, we identified a missense polymorphism in iron-absorption gene CYBRD1 that associated with disease in individuals of English, but not Scottish, ancestry. The association replicated in two additional, independently-collected data sets. Our method can be used to detect associations between genetic variants and disease that have been obscured by population genetic heterogeneity. It can be readily extended to the identification of genetic interactions on other covariates such as measured environmental exposures. We envisage our methodology being of particular interest to researchers with existing GWAS data, as ancestry groups can be easily defined and thus tested for interactions.


Examination of autoantibodies to type I interferon in patients suffering from long COVID.
Long COVID (LC) is an emerging global health concern. The underlying mechanism and pathophysiology remain unclear. Presence of neutralizing autoantibodies against type 1 interferons (IFN) has been established as a predictor of critical COVID-19. We hypothesized that persistent autoimmune activity with autoantibodies against type 1 IFN may contribute to symptoms in patients with LC. Plasma samples and clinical information were obtained from a Danish LC cohort consisting of adult patients with confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Information on symptoms and quality of life was derived from an LC-specific questionnaire and the EQ-5D-5L questionnaire. Detection of type 1 IFN autoantibodies in plasma were performed by ELISA. Samples collected between June, 2020, and September, 2021, from 279 patients were analyzed and compared to a control group of 94 individuals with prior mild SARS-CoV-2 infection who did not develop LC symptoms. In total, five LC patients (1.8%) and 3 (3.2%) of the controls had detectable circulating type 1 IFN autoantibodies. Collectively, prevalence of autoantibodies against type 1 IFN subtypes in our LC cohort were primarily driven by men and did not exceed the prevalence in controls. Thus, in our cohort, anti-type I IFN autoantibodies are unlikely to drive LC symptoms.


Patients with naproxen-induced liver injury display T-cell memory responses toward an oxidative (S)-O-Desmethyl Naproxen metabolite but not the acyl glucuronide.
Exposure to nonsteroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen (IBU) and naproxen (NAP) is associated with idiosyncratic drug-induced liver injury (DILI). Carboxylate bioactivation into reactive metabolites (e.g., acyl glucuronides, AG) and resulting T-cell activation is hypothesized as causal for this adverse event. However, conclusive evidence supporting this is lacking. In this work, we identify CD4+ and CD8+ T-cell hepatic infiltration in a biopsy from an IBU DILI patient. Lymphocyte transformation test and IFN-γ ELIspot, conducted on peripheral blood mononuclear cells (PBMCs) of patients with NAP-DILI, were used to explore drug-specific T-cell activation. T-cell clones (TCC) were generated and tested for drug specificity, phenotype/function, and pathways of T-cell activation. Cells were exposed to NAP, its oxidative metabolite 6-O-desmethyl NAP (DM-NAP), its AG or synthesized NAP-AG human-serum albumin adducts (NAP-AG adduct). CD4+ and CD8+ T-cells from patients expressing a range of different Vβ receptors were stimulated to proliferate and secrete IFN-γ and IL-22 when exposed to DM-NAP, but not NAP, NAP-AG or the NAP-AG adduct. Activation of the CD4+ TCC was HLA-DQ-restricted and dependent on antigen presenting cells (APC); most TCC were activated with DM-NAP-pulsed APC, while fixation of APC blocked the T-cell response. Cross-reactivity was not observed with structurally-related drugs. Our results confirm hepatic T-cell infiltrations in NSAID-induced DILI, and show a T-cell memory response toward DM-NAP indicating an immune-mediated basis for the adverse event. Whilst bioactivation at the carboxylate group is widely hypothesized to be pathogenic for NSAID associated DILI, we found no evidence of this with NAP.


Pregnancy Outcomes in Isolated Oligohydramnios during Second Trimester: A Case Series.
Oligohydramnios is associated with increased maternal and foetal morbidities. However, some of the recent studies have shown no adverse effect of isolated oligohydramnios on perinatal outcome and recommends continuation of pregnancy. Pregnancies between 18-28 weeks with isolated Oligohydramnios were included. History and physical examination was recorded in a pre-designed proforma. All the cases received care as per the protocol. A total of seven patients were recruited of which one was a twin pregnancy with Oligohydramnios in both sacs. The mean age at presentation was 30 years. Three patients went into spontaneous explusion at an average gestational age of 22-24 weeks. One patient with twins delivered vaginally at 32 weeks. Rest were delivered by caesarean section between 34-35 weeks (indication in majority of the cases was foetal distress and cord compression). None of the babies suffered any complication and were discharged in good condition. Isolated oligohydramnios during second trimester does not increase adverse perinatal outcome significantly (but increases the caesarean section rate) and therefore, should not be an indication for termination of pregnancy.


Rationale and design of the Primary pREvention strategies at the community level to Promote Adherence of treatments to pREvent cardiovascular diseases trial number (CTRI/2012/09/002981).
Cardiovascular diseases (CVDs) are a leading cause of morbidity and mortality in low-income countries including India. There is a need for effective, low-cost methods to prevent CVDs in rural India. One strategy is to identify and implement interventions at high-risk individuals using community health workers (CHWs). There is a paucity of CHW-based CVD intervention trials from low-income countries. We designed a multicenter, household-level, cluster-randomized trial with 1:1 allocation to intervention and control arms. The CHWs undertook a door-to-door survey and screened 5,699 households in 28 villages from 3 rural regions in India to identify at-risk households. The households were defined as those with ≥1 individual aged ≥35 years and at moderate or high risk for CVD based on the non-laboratory-based National Health and Nutrition Examination Survey score. All at-risk individuals were invited to attend a physician-led village clinic that provided a CVD risk reduction prescription and education about target risk factor levels for CVD control. All households in which at least 1 member at moderate to high risk for CVD had received a risk reduction prescription were eligible for randomization. Households randomized to the CHW-based intervention will receive 1 household visit by a CHW every 2 months, for 12 months. During these visits, CHWs will measure blood pressure, ascertain and reinforce adherence to prescribed therapies, and modify therapy to meet targets. Households randomized to the control arm do not receive CHW visits. At 12 months after randomization, we will evaluate 2 primary outcomes of systolic blood pressure and adherence to antihypertensive drugs and secondary outcomes of INTERHEART risk score, body mass index, and waist-to-hip ratios. At 18 to 24 months after randomization and 6 to 12 months after the last intervention, we will record these outcomes to evaluate sustainability of intervention. Community health workers screened a total of 5,033 households that included 9,248 individuals and identified 2,571 households with 3,784 at-risk individuals. We randomized 2,438 households (1,219 to intervention and 1,219 to control groups). Our large trial of CHWs in rural India will provide important information regarding a promising approach to primary prevention of CVDs.


Modulation of the digestive lysosomal system in Paramecium caudatum. III. Morphological effects of cytochalasin B.
Morphological explanations for the effects of cytochalasin B (CB) on the digestive vacuole system of Paramecium caudatum were sought using electron-microscopic studies. Cytochalasin B (0.3 mM) essentially stopped vacuole release without stopping vacuole growth. Acidosome fusion with the new vacuoles was inhibited and was accompanied by the formation of discoidal vesicle-lined clumps of microfilament-like material next to these vacuoles. Vacuole membrane retrieval at the open cytoproct was inhibited and was accompanied by the loss of the microfilament network typical of this region. At 0.15 mM CB vacuole release and fusion of the acidosomes and lysosomes with the vacuoles were not stopped, but membrane retrieval at the cytoproct was affected. Cytochalasin B at the higher concentration also affected the movement of the horseradish peroxidase (HRP)-labeled vesicles away from the nascent vacuole. Membrane retrieval by the formation of tubules of 35 to 50 nm diameter from membrane of early and late vacuole stages was also affected by CB. It is conceivable that all of these effects were the result of the disruption of membrane associated actin networks. By contrast, several highly organized microfilamentous systems in the cortex and oral region of these cells appeared to be unaffected by CB or dimethyl sulfoxide (DMSO) at the concentrations used. These results show that CB-susceptible structures assume a key role in the normal functioning of the digestive vacuole system and in vacuole membrane retrieval and reutilization in Paramecium.


Altered stationary-phase response in a Borrelia burgdorferi rpoS mutant.
The homolog of the chromosomally encoded stationary-phase sigma factor RpoS in Borrelia burgdorferi was inactivated using gyrB(r) as a selectable marker. Two-dimensional nonequilibrium pH gradient electrophoresis of stationary-phase cell lysates identified at least 11 differences between the protein profiles of the rpoS mutant and wild-type organisms. Wild-type B. burgdorferi had a growth phase-dependent resistance to 1 N NaCl, similar to the stationary-phase response reported for other bacteria. The B. burgdorferi rpoS mutant strain was less resistant to osmotic stress in stationary phase than the isogenic rpoS wild-type organism. The results indicate that the B. burgdorferi rpoS homolog influences protein composition and participates in stationary-phase-dependent osmotic resistance. This rpoS mutant will be useful for studying regulation of gene expression in response to changing environmental conditions.


Mechanism for cyclization reaction by clavaminic acid synthase. Insights from modeling studies.
The mechanism of the oxidative cyclization reaction catalyzed by clavaminic acid synthase (CAS) was studied in silico. First, a classical molecular dynamics (MD) simulation was performed to obtain a realistic structure of the CAS-Fe(IV)=O-succinate-substrate complex; then potential of mean force (PMF) was calculated to assess the feasibility of the beta-lactam ring, more specifically its C4' corner, approaching the oxo atom. Based on the MD structure, a relatively large model of the active site region was selected and used in the B3LYP investigation of the reaction mechanism. The computational results suggest that once the oxoferryl species is formed, the oxidative cyclization catalyzed by CAS most likely involves either a mechanism involving C4'(S)-H bond cleavage of the monocyclic beta-lactam ring, or a biosynthetically unprecedented mechanism comprising (1) oxidation of the hydroxyl group of PCA to an O-radical, (2) retro-aldol-like decomposition of the O-radical to an aldehyde and a C-centered radical, which is stabilized by the captodative effect, (3) abstraction of a hydrogen atom from the C4'(S) position of the C-centered radical by the Fe(III)-OH species yielding an azomethine ylide, and (4) 1,3-dipolar cycloaddition to the ylide with aldehyde acting as a dipolarophile. Precedent for the new proposed mechanism comes from the reported synthesis of oxapenams via 1,3-dipolar cycloaddition reactions of aldehydes and ketones.


Ethanol inhibits muscarinic receptor-stimulated phosphoinositide metabolism and calcium mobilization in rat primary cortical cultures.
In recent years, it has been hypothesized that muscarinic receptor-stimulated phosphoinositide (PI) metabolism may represent a relevant target for the developmental neurotoxicity of ethanol. Age-, brain region-, and receptor-specific inhibitory effects of ethanol on this system have been found, both in vitro and after in vivo administration. As a direct consequence of this action, alterations of calcium homeostasis would be expected, through alterations of inositol trisphosphate formation, which mediates intracellular calcium mobilization. In the present study, the effects of ethanol (50-500 mM) on carbachol-stimulated PI metabolism and free intracellular calcium levels were investigated in rat primary cortical cultures, by measuring release of inositol phosphates and utilizing the two calcium probes fluo-3 and indo-1 on an ACAS (Adherent Cell Analysis and Sorting) Laser Cytometer. Ethanol exerted a concentration-dependent inhibition of carbachol-stimulated PI metabolism. In addition, ethanol's inhibitory effect paralleled the temporal development of the muscarinic receptor signal transduction system, with the strongest inhibition (25-50%) occurring when maximal stimulation by carbachol occurs (days 5-7). Ethanol also exerted a concentration-dependent decrease in free intracellular calcium levels following carbachol stimulation. Both initial calcium spike amplitude, seen in all responsive cells, as well as the total number of cells responding to carbachol, were decreased by ethanol. The inhibitory effects of ethanol seemed dependent upon preincubation time, in that a longer preincubation (30 min) with the lowest dose (50 mM), showed almost the same decrease in responding cell number and reduction in spike amplitude in responding cells, as a shorter incubation (10 min) with the highest ethanol dose (500 mM). The specificity of the response to carbachol was demonstrated by blocking the response with 10 microM atropine. Moreover, experiments with carbachol in calcium-free buffer with 1 mM EGTA indicated that the initial calcium spike was due to intracellular calcium mobilization from intracellular stores. Since calcium is believed to play important roles in cell proliferation and differentiation, these results support the hypothesis that this intracellular signal-transduction pathway may be a target for ethanol, contributing to its developmental neurotoxicity.


Inactivation of the alpha C protein antigen gene, bca, by a novel shuttle/suicide vector results in attenuation of virulence and immunity in group B Streptococcus.
The alpha C protein of group B Streptococcus (GBS) is a major surface-associated antigen. Although its role in the biology and virulence of GBS has not been defined, it is opsonic and capable of eliciting protective immunity. The alpha C protein is widely distributed among clinical isolates and is a potential protein carrier and antigen in conjugate vaccines to prevent GBS infections. The structural gene for the alpha C protein, bca, has been cloned and sequenced. The protein encoded by bca is related to a class of surface-associated proteins of gram-positive cocci involved in virulence and immunity. To investigate the potential roles of the alpha C protein, bca null mutants were generated in which the bca gene was replaced with a kanamycin resistance cassette via homologous recombination using a novel shuttle/suicide vector. Studies of lethality in neonatal mice showed that the virulence of the bca null mutants was attenuated 5- to 7-fold when compared with the isogenic wild-type strain A909. Significant differences in mortality occurred in the first 24 h, suggesting that the role of the alpha antigen is important in the initial stages of the infection. In contrast to A909, bca mutants were no longer killed by polymorphonuclear leukocytes in the presence of alpha-specific antibodies in an in vitro opsonophagocytic assay. In contrast to previous studies, alpha antigen expression does not appear to play a role in resistance to opsonophagocytosis in the absence of alpha-specific antibodies. In addition, antibodies to the alpha C protein did not passively protect neonatal mice from lethal challenge with bca mutants, suggesting that these epitopes are uniquely present within the alpha antigen as expressed from the bca gene. Therefore, the alpha C protein is important in the pathogenesis of GBS infection and is a target for protective immunity in the development of GBS vaccines.


Cleaning and sterilisation of infant feeding equipment: a systematic review.
To assess the clinical and cost-effectiveness of different methods of cleaning and sterilisation of infant feeding equipment used in the home. Systematic review of studies from developed countries on the effectiveness of methods of cleaning and sterilisation of infant feeding equipment used in the home. A brief telephone survey of UK-based manufacturers of infant feeding equipment and formula to ascertain the evidence base used for their recommendations, and a comparison of current relevant guidelines in developed countries, informed the work. National guidelines from six countries demonstrated variation and lack of evidence to support current guidance. Manufacturers did not report evidence of effectiveness to support their recommendations. Nine studies were identified; eight conducted between 1962 and 1985 and one in 1997. All had methodological weaknesses. Hand-washing was identified as fundamentally important. Health professionals were reported as not providing appropriate education on the importance and methods of cleaning and sterilisation. Mothers of subsequent babies and women from lower socio-economic groups were less likely to follow recommended procedures. There is a lack of good-quality evidence on effective ways of cleaning and sterilising infant feeding equipment in the home. The evidence base does not answer the question about which of the methods in common use is most effective or most likely to be used by parents. Hand-washing before handling feeding equipment remains important. Further research on the range of methods used in the home environment, including assessment of the views of parents and carers, is required.


DPP4 Inhibitors Can Be a Drug of Choice for Type 3 Diabetes: A Mini Review.
As well known to the scientific community, Alzheimer's disease (AD) is an irreversible neurodegenerative disease that ends up with impairment of memory and cognition due to neuronal and synapse loss. Patient's quality of life can be enhanced by targeting neurogenesis as a therapeutic paradigm. Moreover, several research evidences support the concept that AD is a type of metabolic disorder mediated by impairment in brain insulin responsiveness and energy metabolism. Growing evidence suggests that endogenous peptides such as glucagon-like peptide-1 (GLP-1) and stromal-derived factor-1α (SDF-1α) provide neuroprotection across a range of experimental models of AD. So, preserving functional activity of SDF-1α and GLP-1 by dipeptidyl peptidase-4 inhibition will enhance the homing/recruitment of brain resident and nonresident circulating stem cells/progenitor cells, a noninvasive approach for promoting neurogenesis. So, herewith we provide this in support of dipeptidyl peptidase-4 inhibitors as a new target of attention for treating AD.


Racial and ethnic differences in the association between pregnancy dysglycemia and cardiometabolic risk factors 10-14 years' postpartum in the HAPO follow-up study.
Associations between pregnancy dysglycemia and subsequent maternal cardiometabolic factors 10-14 years postpartum were largely similar across self-identified racial and ethnic groups among birthing people in the U.S. enrolled in the Hyperglycemia and Adverse Pregnancy Outcome (HAPO) Follow-up Study.


B cell clones that sustain long-term plasmablast growth in T-independent extrafollicular antibody responses.
Some antigens induce Ab responses without T lymphocyte help. Among these, many polysaccharide-based antigens cause marginal zone B cells to proliferate and differentiate into plasma cells. B1 cells also respond to some of these antigens. In this article, we report that antigen-specific B1b cells, in response to the T-independent antigen (4-hydroxy-3-nitrophenyl)-acetyl (NP)-Ficoll, develop into clones that sustain Ab production for months with continued production of plasma cells and the accumulation of antigen-specific B cells in follicles. The persistence of this T-independent plasmablast response contrasts with the short-term plasmablast growth associated with T-dependent extrafollicular responses. The nature of the cells responding to NP-Ficoll was probed by using chimeras that have B1 cells but lack primary B lymphopoietic capacity and have very few B2 cells or T cells. The chimeras were constructed by transferring 10(5) IgM(+) IgD(-) peritoneal exudate cells into mice unable to produce their own T and B cells because of deficiency in recombinase-activating gene 1 (RAG-1). The chimeras mounted sustained IgM and IgG3 anti-NP Ab responses to NP-Ficoll. This finding was associated with continued NP-specific extrafollicular plasmablast growth and the accumulation of NP-specific B cells in follicles. B cells were not found in the marginal zones of chimeras, and they also lacked recirculating IgD(+) cells and CD3(+) cells. The absence of B2 and T cells confirms that hemopoietic cell chimerism leading to primary lymphopoiesis had not been established.


Prokaryote-eukaryote relationship and the amino acid sequence of plastocyanin from Anabaena variabilis.
The amino acid sequence of plastocyanin from the prokaryotic blue-green alga Anabaena variabilis was determined. The protein consists of a single polypeptide chain of 105 residues. The amino acid sequence of the plastocyanin was compared with that of the eukaryotic green alga Chlorella fusca and with those of higher-plant plastocyanins. The considerable similarity between the prokaryotic and eukaryotic plastocyanins is discussed. Detailed evidence for the sequence of the protein has been deposited as Supplementary Publication SUP 50051 (13 pages) at the British Library (Lending Division), Boston Spa, Wetherby, W. Yorkshire LS23 7BQ, U.K., from whom copies may be obtained on the terms given in Biochem J. (1975) 145, 5.


Adaptive observer backstepping control using neural networks.
This paper extends the application of neurocontrol approaches to a new class of nonlinear systems diffeomorphic to output feedback nonlinear systems with unmeasured states. A neural-based adaptive observer is introduced for state estimation as well as system identification using only output measurements during online operation. System identification is achieved via the online approximation of a priori unknown functions. The controller is designed using the backstepping control design procedure. Leakage terms in the adaptive laws and nonlinear damping terms in the backstepping controller are introduced to prevent instability from arising due to the inherent approximation error. A primary benefit of the online function approximation is the reduction of approximation errors, which allows reduction of both the observer and controller gains. A semi-global stability analysis for the proposed approach is provided and the feasibility is investigated by an illustrative simulation example.


The 80-hour work week for residents: views from obstetric and gynecology program directors.
In 2003, the Accreditation Council for Graduate Medical Education mandated an 80-hour work week restriction for residency programs. We examined program directors' views on how this mandate affects the education of Obstetrics and Gynecology residents. A 25 question survey was administered via Survey Monkey to Obstetrics and Gynecology program directors in the United States over three months in 2011. Fifty program directors (response rate of 28%) completed it with more men (62%) than women (38%) respondents. Overall, only 28% (14/50) responded that the program had improved, with significantly fewer men (5/14; 16.1%) than women (47.4% 9/19; p < 0.0169) directors reporting this. There was little perceived improvement in any of the six core ACGME performance objectives and in the CREOG scores, with the improvement ranging from 8% to 12%. In fact, while we observed the percentage of women directors reporting improvement in patient care and interpersonal and communication skills significantly higher compared with their male counterparts, the majority of women still reported either no improvement or a decline in these areas. Though our sample size was small, we found some significant difference between the views of male and female program directors. Both groups nonetheless responded with the majority with a decline or no change rather than a perceived improvement in any of the educational endeavors studied.


Chronology, dynamics and period of permanent tooth eruption in Zagreb children (Part II).
The aim of the present paper is to determine the beginning, order, median time, termination, dynamics, continuity and intensity of the permanent tooth eruption, as well as its calendar, in a sample of Zagreb children. The study included 2768 children (1398 boys and 1370 girls). All subjects were classified in one-year age groups. The continuity of tooth eruption occurs from 5.1 years to 14.97 years, i.e., for a period of 9.87 years. Each particular tooth has its own dynamics of eruption (V5-V95) which on the average amounts to 4.25 years. The continuity of eruption is 10.1 years in boys and 9.8 years in girls. The dynamics of eruption is 4.15 years in boys and 4.35 years in girls. A certain asymmetry in eruption has been described.


Urea-induced denaturation of beta-trypsin: an evidence for a molten globule state.
The denaturation of beta-trypsin induced by urea was investigated by fluorescence and circular dichroism. A transient denatured state was found at 2 M urea in both intrinsic fluorescence spectrum and bis-(8-anilino-1-naphtalene sulfonate) (bis-ANS) binding. In addition, the absence of tertiary contacts and presence of secondary structure for this state, are consistent with an intermediate equilibrium state having features of molten globule.


Miconazole therapy for endemic fungal disease.
Miconazole nitrate (2.0%) cream was evaluated in the treatment of endemic dermatophytosis in an overcrowded prison. The active drug was compared in a random double-blind study with its control vehicle in the treatment of 99 inmates who had moderate to severe symptomatic disease. Tinea cruris and tinea pedis caused by Trichophyton rubrum predominated, although Trichophyton mentagrophytes and Candida albicans were also isolated and identified. Prompt clinical clearing occurred in 75.5% of those treated with miconazole versus 12.0% of those treated with the vehicle alone. Results of mycological examination closely paralleled and confirmed these clinical results.


Cytokeratin expression in smooth muscle and smooth muscle tumours.
The expression of cytokeratin intermediate filaments by a tumour has been accepted as evidence of an epithelial origin. Although there have been anecdotal reports of cytokeratin expression within tissues and neoplasms of non-epithelial origin, particularly muscle, there have been no comprehensive studies of its frequency and distribution. In order to investigate this we have studied 51 cases of normal smooth muscle and benign and malignant smooth muscle tumours using a panel of monoclonal antibodies against a range of intermediate filaments (cytokeratins, desmin and vimentin). Cytokeratin expression was noted overall in 50% of normal, benign and malignant smooth muscle tissues. Such expression tended to have a focal or patchy distribution. No case expressed cytokeratins in the absence of both desmin and vimentin. The implication of these findings for diagnostic immunocytochemistry is that intermediate filaments alone are not completely reliable markers of tumour histogenesis and should be used as part of a larger panel of monoclonal antibodies.


Directed evolution of AraC for improved compatibility of arabinose- and lactose-inducible promoters.
Synthetic biological systems often require multiple, independently inducible promoters in order to control the expression levels of several genes; however, cross talk between the promoters limits this ability. Here, we demonstrate the directed evolution of AraC to construct an arabinose-inducible (P(BAD)) system that is more compatible with IPTG (isopropyl-beta-D-1-thiogalactopyranoside) induction of a lactose-inducible (P(lac)) system. The constructed system is 10 times more sensitive to arabinose and tolerates IPTG significantly better than the wild type. Detailed studies indicate that the AraC dimerization domain and C terminus are important for the increased sensitivity of AraC to arabinose.


[The role of insects (Blattodea, Diptera, and Hymenoptera) as possible mechanical vectors of helminths in the domiciliary and peridomiciliary environment].
Helminths can be transmitted to human beings in several ways, but little attention has been given to vector or mechanical transmission of infective forms by insects. The present study surveys the helminth species present in three orders of insects that coexist in proximity with the human environment. A total of 700 insects (54 Blattodea, 275 Diptera, and 371 Hymenoptera) were collected and examined externally and individually. In the Blattodea order, only specimens of Periplaneta americana were collected, and 58.3% were carrying the following helminth forms: Oxyuridae eggs (36.4%), Ascaridae eggs (28.04%), Nematoda larvae (4.8%), Cestoda eggs (3.5%), other Nematoda (0.08%), and Toxocaridae eggs (0.08%). No Diptera and Hymenoptera were found to contain parasitic forms. This study evaluates the importance and role of insects as mechanical vectors of helminth parasites, correlated with social and environmental conditions, and suggests the use of these data for preventive purposes.


Regulation of the action of hydrocortisone in airway epithelial cells by 11beta-hydroxysteroid dehydrogenase.
11beta-hydroxysteroid dehydrogenase (11betaHSD) reversibly converts hydrocortisone, the predominant active endogenous glucocorticoid in humans, to its inactive metabolite cortisone by oxidizing the 11-hydroxy group to an 11-keto group. Because this enzyme is highly expressed in human bronchial epithelial cells, we hypothesized that it regulates epithelial responses to glucocorticoids by reducing levels of hydrocortisone available to bind to the glucocorticoid receptor. Primary human bronchial epithelial cells (PBECs) were isolated from seven autopsy specimens and cultured in F12/Dulbecco's modified Eagle's medium with 5% fetal bovine serum until approximately 80% confluent. Cells were preincubated with 10(-9) M to 10(-5) M hydrocortisone for 24 h in the presence or absence of 10(-6) M of the 11betaHSD inhibitor glycyrrhetinic acid, after which the cells were stimulated with 5 ng/ml interleukin-1beta for 24 h. Granulocyte macrophage colony-stimulating factor (GM-CSF) levels were quantitated in the resulting supernatants by enzyme-linked immunosorbent assay. Hydrocortisone inhibited GM-CSF release in stimulated PBEC with a concentration that produces 50% inhibition of maximum effect (IC(1/2)max) of 5.0 x 10(-8) M. In the presence of glycyrrhetinic acid, the potency of hydrocortisone was increased approximately 33-fold (IC(1/2)max with glycyrrhetinic acid, 1.5 x 10(-9) M). Hydrocortisone activity was maximally enhanced at concentrations between 10(-9) M and 10(-8) M, levels that are comparable to plasma levels of hydrocortisone not bound to plasma proteins. Glycyrrhetinic acid had no effect on the suppression of GM-CSF release by hydrocortisone in the transformed cell line BEAS-2B, which does not express the 11betaHSD enzyme. Glycyrrhetinic acid also had no effect on the inhibition of GM-CSF release in PBECs by the synthetic glucocorticoids budesonide, beclomethasone dipropionate, fluticasone propionate, mometasone furoate, and triamcinolone acetonide, steroids not metabolized by 11betaHSD. Together, these findings suggest that metabolism of hydrocortisone by 11betaHSD may regulate glucocorticoid activity in human airway epithelial cells.


Systematic Review of Three Electrical Stimulation Techniques for Rehabilitation After Total Knee Arthroplasty.
The comparative effectiveness of neuromuscular electrical stimulation (NMES), transcutaneous electrical nerve stimulation (TENS), and electroacupuncture (EA) for improving patient rehabilitation following total knee arthroplasty (TKA) is controversial. Therefore we conducted this systematic review to assess the available evidence. The PubMed, OVID, and ScienceDirect databases were comprehensively searched and studies were selected and analyzed according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) recommendations. Data were extracted and qualitatively synthesized for several outcomes. Data were analyzed from 17 randomized controlled trials involving 1285 procedures: 8 NMES studies (608 procedures), 7 TENS studies (560 procedures), and 2 EA studies (117 procedures). Qualitative analysis suggested that NMES was associated with higher quadriceps strength and functional recovery after TKA. Recovery benefits were maximal when the stimulation was performed once or twice a day for 4-6 weeks at an intensity of 100-120 mA and frequency of 30-100 Hz. The electrode should be sufficiently large (100-200 cm2) to reduce discomfort. TENS at an intensity of 15-40 mA and frequency of 70-150 Hz provided effective analgesia after TKA. EA at an intensity of 2 mA and frequency of 2 Hz may also provide postoperative analgesia of TKA. As adjunct modalities, NMES and TENS can effectively improve rehabilitation after TKA without triggering significant intolerance, and maximal benefits depend on optimized parameters and intervention protocols. EA may be an effective adjunct modality for analgesia after TKA.


The microglial NLRP3 inflammasome is activated by amyotrophic lateral sclerosis proteins.
Microglial NLRP3 inflammasome activation is emerging as a key contributor to neuroinflammation during neurodegeneration. Pathogenic protein aggregates such as β-amyloid and α-synuclein trigger microglial NLRP3 activation, leading to caspase-1 activation and IL-1β secretion. Both caspase-1 and IL-1β contribute to disease progression in the mouse SOD1G93A model of amyotrophic lateral sclerosis (ALS), suggesting a role for microglial NLRP3. Prior studies, however, suggested SOD1G93A mice microglia do not express NLRP3, and SOD1G93A protein generated IL-1β in microglia independent to NLRP3. Here, we demonstrate using Nlrp3-GFP gene knock-in mice that microglia express NLRP3 in SOD1G93A mice. We show that both aggregated and soluble SOD1G93A activates inflammasome in primary mouse microglia leading caspase-1 and IL-1β cleavage, ASC speck formation, and the secretion of IL-1β in a dose- and time-dependent manner. Importantly, SOD1G93A was unable to induce IL-1β secretion from microglia deficient for Nlrp3, or pretreated with the specific NLRP3 inhibitor MCC950, confirming NLRP3 as the key inflammasome complex mediating SOD1-induced microglial IL-1β secretion. Microglial NLRP3 upregulation was also observed in the TDP-43Q331K ALS mouse model, and TDP-43 wild-type and mutant proteins could also activate microglial inflammasomes in a NLRP3-dependent manner. Mechanistically, we identified the generation of reactive oxygen species and ATP as key events required for SOD1G93A -mediated NLRP3 activation. Taken together, our data demonstrate that ALS microglia express NLRP3, and that pathological ALS proteins activate the microglial NLRP3 inflammasome. NLRP3 inhibition may therefore be a potential therapeutic approach to arrest microglial neuroinflammation and ALS disease progression.


Maternally inherited X chromosome is not inactivated in mouse blastocysts due to parental imprinting.
Mouse embryos having an additional maternally inherited X chromosome (X(M)) invariably die before midgestation with the deficient extraembryonic ectoderm of the polar trophectoderm lineage, whereas postnatal mice having an additional paternally inherited X chromosome (X(P)) survive beyond parturition. A cytogenetic study led us to hypothesize that abnormal development of such embryos disomic for X(M) (DsX(M)) is attributable to two doses of active X(M) chromosome in extraembryonic tissues. To test the validity of this hypothesis, we examined the initial X chromosome inactivation pattern in embryos at the blastocyst stage by means of replication banding method as well as RNA FISH detecting Xist transcripts. X(P) was the only asynchronously replicating X chromosome, if any, in X(M)X(M)X(P) blastocysts, and no such allocyclic X chromosome was ever detected in X(M)X(M)Y blastocysts. In agreement with these findings, only one Xist paint signal was detected in 79% of X(M)X(M)X(P) cells, whereas no such signal was found in X(M)X(M)Y embryos. Thus, the present study supports the hypothesis that two X chromosomes remaining active in the extraembryonic cell lineages due to the maternal imprinting explain the underdevelopment of extraembryonic structures and hence early postimplantation death of DsX(M) embryos.


Correlates of smoking among youth: the role of parents, friends, attitudes/beliefs, and demographics.
Family engagement has been shown to play a crucial role in youth cigarette use prevention and uptake. We examine cross-sectional and longitudinal data to determine whether changes in parental monitoring factors influence changes in smoking susceptibility. Two cross-sectional surveys of Florida youth (12-17 years) were conducted in 2009, with a follow-up survey in 2010. Multivariable analyses examined demographics, parent characteristics, family engagement, and parental monitoring on youth susceptibility to smoke. Cross-sectional data show eating together 6+ times/week and doing something for fun 5+ times/week were related to an increased likelihood of Very Low and decreased likelihood of High susceptibility, respectively. Parental monitoring factors and parents tell on a friend who smokes was significantly related to having Very Low susceptibility in both surveys. Mother's education, parent smokes, family engagement factors, and parental monitoring were significant in both survey rounds. Longitudinal analyses showed change in eating together did not significantly affect the odds of change in smoking susceptibility; however, change in the frequency of doing things for fun with a parent showed decreased odds of susceptibility (OR = .63 [.49-.82]), opposite of the hypothesized direction. Lastly, as youth aged, they were more likely to experience a greater odds of decreased susceptibility (OR14-15y = 1.47 [1.08-1.99] and OR≥16y = 1.40 [1.05-1.84], respectively) and less likely to experience an increased odds of susceptibility (OR14-15y = .65 [.49-.86] and OR≥16y = .72 [.56-.93], respectively). We found mixed results for family engagement and parental monitoring on changes in youth smoking susceptibility. Cross-sectional data showed general associations in the expected direction; however, longitudinal analyses showed family engagement variables had significance, but in the opposite hypothesized direction.


HGF and c-MET as potential orchestrators of invasive growth in head and neck squamous cell carcinoma.
Head and neck squamous cell carcinoma (HNSCC) constitutes 90% of all head and neck cancers and is associated with high mortality rates, due to the high infiltrative potential of these tumors. Despite advances in treatment approaches, there has been no improvement in survival rates. As empiricism in the treatment of HNSCC is unlikely to improve the prognosis of HNSCC patients, understanding the pathogenesis behind the local invasion of these tumors on the cellular and molecular levels has become an important goal in the field of head and neck surgery. It is believed that the invasive growth of neoplastic cells is a deregulated form of the physiological program that occurs during the formation and patterning of an embryo, which is largely facilitated by HGF induced signaling through its receptor c-MET. This review investigates whether HGF and c-MET are deregulated in HNSCC and whether they confer an invasive potential to these tumors. It is concluded that both molecules are mis- and over-expressed in HNSCC and probably induce the program of invasive growth in HNSCC.


Safety and efficacy of extracorporeal shock wave therapy for chronic lateral epicondylitis.
Efficacy and safety of extracorporeal shock wave therapy (ESWT) were investigated in 36 patients with chronic lateral epicondylitis--9 patients receiving and 27 patients not receiving worker's compensation. All patients were treated with a single application of 3200 shock waves. Twelve weeks after treatment, the mean visual analog scale score for the entire group improved from 8.0 to 2.5 (P < .05), and the mean RAND 36-Item Health Survey (Physical Functioning) score improved from 65.6 to 88.0 (P < .05). Outcomes for 28 elbows (77.8%) were rated excellent or good on the Roles and Maudsley scale. There were no significant differences in outcome measures among the subgroups. There were no significant complications. ESWT is an effective treatment for chronic lateral epicondylitis. Worker's compensation status did not affect outcomes.


Responses of carotid artery in mice deficient in expression of the gene for endothelial NO synthase.
We examined the hypotheses that responses to acetylcholine are impaired and responses to NO are enhanced in carotid artery from mice made deficient in endothelial nitric oxide synthase (eNOS) by gene targeting (eNOS-deficient mice). We also tested the hypothesis that deletion of one copy of the eNOS gene is sufficient to alter vascular responses. Vessels were studied in vitro from heterozygous (+/-) and homozygous (-/-) eNOS-deficient mice as well as wild-type [eNOS(+/+)] littermates. After precontraction with prostaglandin F2 alpha, acetylcholine produced marked relaxation of carotid arteries in eNOS(+/+) mice, with impaired vasorelaxation in eNOS(+/-) mice. For example, 1 microM acetylcholine relaxed carotid arteries by 55 +/- 5% (mean +/- SE) in eNOS(+/-) mice (n = 13) compared with 83 +/- 3% in eNOS(+/+) mice (n = 14, P < 0.001 vs. +/-). In contrast, acetylcholine caused no relaxation in carotid arteries from eNOS(-/-) mice (P < 0.001 vs. +/+ and +/-). Relaxation of the carotid artery in response to nitroprusside [a nitric oxide (NO) donor] was enhanced (P < 0.001) in eNOS-deficient mice. For example, in response to 10 nM nitroprusside, the carotid artery relaxed by 18 +/- 2% in eNOS(+/+) mice (n = 14), 33 +/- 2% in eNOS(+/-) mice (n = 13), and 47 +/- 4% in eNOS(-/-) mice (n = 5). Thus relaxation of the carotid artery is impaired with acetylcholine and enhanced with the NO donor nitroprusside in eNOS-deficient mice. Enhanced responses to NO may represent a compensatory response expressed in the absence of eNOS. The findings that vascular responses to acetylcholine and NO are altered in eNOS(+/-) mice compared with those observed in eNOS(+/+) mice suggest a "gene-dosing" effect.


[Acute coronary syndrome. Physiopathology and genetics].
The acute coronary syndrome (ACS) is characterized by a spectrum of arterial diseases that include unstable angina and myocardial infarction. In the last 10 years, ACS has become the cause of up to 29% of deaths in the industrialized countries, becoming the main cause of death, and it will most probably stay that way for the year 2020. The physiopathogenesis of ACS include oxidative, inflammatory, and thrombotic processes. Diverse molecules participate in the processes, increasing of decreasing the damage. The genes that encode these molecules have been associated with the disease. However, in some cases inconsistent results in different populations have been reported. In this review the physiopathogenesis and the role of several candidate genes involved in the pathogenesis of ACS are discussed.


Fabrication of a biochar-doped monolithic adsorbent and its application for the extraction and determination of coumarins from Angelicae Pubescentis Radix.
A monolithic adsorbent was designed aiming to the structure of osthole and columbianadin, and fabricated using diallyl phthalate as the monomer and ethylene dimethacrylate as the crosslinker with the addition of bamboo biochar, via polymerization reaction in a stainless-steel tube. The prepared composite adsorbent packed in the tube was used as a solid-phase extraction column for the extraction and determination of two coumarins (osthole and columbianadin) in Angelicae Pubescentis Radix, combing with a C18 analytical column through an HPLC instrument, which show excellent matrix-removal ability and good selectivity to osthole and columbianadin. Furthermore, the present adsorbent shows good applicability, which was used for the extraction of osthole from Duhuo Jisheng Pill. Compared to the commercial C18 and phenyl adsorbent, the present adsorbent own better selectivity and higher resolution. These results attributed to the enhanced specific surface area (141 m2/g) and enriched interaction sites of the resulting composite adsorbent, due to the doping of bamboo biochar, which can produce hydrogen bond, dipole-dipole, π-π and hydrophobic force interactions with the osthole and columbianadin. The methodology validation indicated that the present method showed good precision and good accuracy, and the composite adsorbent showed good preparative repeatability, which can be reused for no less than 100 times with the relative standard deviation ≤4.6 % (n = 100). The present work provided a simple and efficient method for the extraction and determination osthole and columbianadin from Angelicae Pubescentis Radix.


Return to Sports After Athletes Undergo Meniscal Surgery: A Systematic Review.
The aim of this systematic review was to determine the time to and rate of the return to sports (RTS) after meniscal surgery and to compare these values among the different types of meniscal surgeries. Systematic review. The MEDLINE, EMBASE, and Cochrane databases were searched. Studies that met the following criteria were included: (1) English article, (2) full-text available, (3) study on athletes, and (4) primary outcome is RTS after meniscal surgery, such as meniscectomy, meniscal repair, and meniscal allograft transplantation (MAT). Eleven articles were included in the final analysis. Most players returned to the preinjury activity level from 7 to 9 weeks after partial meniscectomy. After meniscal repair, 81% to 88.9% of athletes returned to sports. The time to RTS after isolated meniscal repair was on average 5.6 months, and the time to RTS was longer for athletes who required concurrent anterior cruciate ligament reconstruction (ACLR). After MAT, 67% to 85.7% of athletes returned to sports, and the time to RTS ranged from 7.6 to 16.5 months. The shortest time to RTS and the highest RTS rate were observed after partial meniscectomy. The time to RTS was shorter, and the RTS rate was higher after meniscal repair than after MAT. Concurrent procedures such as ACLR prolonged the time to RTS, but it had no effect on the RTS rate and the level of sports activity at the time of RTS.


How much tubal factor infertility is caused by Chlamydia? Estimates based on serological evidence corrected for sensitivity and specificity.
To estimate the proportion of tubal factor infertility (TFI) caused by Chlamydia trachomatis (CT), the etiologic fraction, from a retrospective study of CT antibody prevalence in TFI cases and controls, adjusted for sensitivity and specificity. We use published data on sensitivity and specificity to estimate the performance of assays in (a) women with a previous CT infection without sequelae and (b) women with TFI caused by CT. A model was developed and applied to antibody prevalence in TFI cases and controls from 1 published case-control study to estimate the proportion of TFI caused by CT. The proportion of TFI episodes that were due to CT infection was estimated to be 45% (credible intervals: 28%, 62%). Models which assume that test sensitivity is higher in women with CT-related TFI than women with previous infection and no sequelae fit the data significantly better than models that assume the same sensitivity in all those previously infected. Greater attention needs to be paid to methods for characterizing the performance of CT antibody tests. Serological studies could be given a greater role both in CT etiology and in monitoring the effects of prevention and control programmes.


Friction on incommensurate substrates: Role of anharmonicity and defects.
We present molecular dynamics simulations of one- and two-dimensional bead-spring models sliding on incommensurate substrates after an initial kick, in the case where the coupling to the underlying substrate is weak, i.e., energy can dissipate only into the internal degrees of freedom of the sliding object, but not into the substrate below. We investigate how sliding friction is affected by structural defects and interaction anharmonicity. In their absence, we confirm earlier findings, namely, that at special resonance sliding velocities, friction is maximal. When sliding off-resonance, partially thermalized states are possible, whereby only a small number of vibrational modes becomes excited, but whose kinetic energies are already Maxwell-Boltzmann distributed. Anharmonicity and defects typically destroy partial thermalization and instead lead to full thermalization, implying much higher friction. For sliders with periodic boundaries, thermalization begins with vibrational modes whose spatial modulation is compatible with the incommensurate lattice. For a disk-shaped slider, modes corresponding to modulations compatible with the slider radius are initially the most dominant. By tuning the mechanical properties of the slider's edge, this effect can be controlled, resulting in significant changes in the sliding distance covered.


Metabotropic glutamate receptor-mediated hippocampal phosphoinositide turnover is blunted in spatial learning-impaired aged rats.
Maximal phosphoinositide (PI) turnover was examined in the hippocampus of young and aged Long-Evans rats that were behaviorally characterized for spatial learning in the Morris water maze. The type 1 metabotropic glutamate receptor (mGluR) agonist 1S,3R ACPD was used to stimulate PI turnover and to determine the E(MAX) for each rat. Protein levels in hippocampus for type 1 mGluRs, Galphaq11, and phospholipase Cbeta-1 (PLCbeta-1) were also measured by quantitative Western blotting. The results show that PI turnover mediated by the mGluRs was blunted in the aged rats. The magnitude of the decrement in PI turnover was also significantly correlated with age-related spatial memory decline. The decrease in mGluR-mediated PI turnover occurred without changes in the protein level of either the mGluRs or the G-protein coupled to those receptors, Galphaq11. A significant decrease in the immunoreactivity of PLCbeta-1, however, was observed in the hippocampus of aged rats; PLCbeta-1 immunoreactivity was significantly correlated with spatial learning only when the young and aged rats were considered together. The decrement in mGluR-mediated signal transduction in the hippocampus that is related to cognitive impairment in aging may be attributable, at least in part, to a deficiency in the enzyme PLCbeta-1. That deficiency may also contribute to a blunted response in muscarinic stimulation of hippocampal PI turnover that we previously found in this same study population. An age-related alteration in this signal transduction system may provide a functional basis for cognitive decline independent of any loss of neurons in the hippocampus.


Longitudinal limb deficiencies and the sclerotomes. An analysis of 378 dysmelic malformations induced by thalidomide.
The pathogenesis of longitudinal reduction deformities of the limbs, or dysmelia, is still a matter of debate. Their morphological pattern was defined from a large collection of radiographs of children with dysmelia following the thalidomide disaster. We compared radiographs of 378 of these limbs with the sclerotomes which are areas of segmental sensory innervation of the limb skeleton defined by the radiation of referred pain. The pattern of dysmelia matched the sclerotomes closely in 279 limbs (73.5%). The principles of skeletal reduction in dysmelia are explained by the arrangement of the sclerotomes. The congruence between two separate and independent data sets shows that both patterns are expressions of the underlying segmental sensory innervation of the skeleton, and that the sensory nervous system is involved in the process of limb morphogenesis and teratogenesis.


Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings.
Multichannel electrode arrays offer insight into the working brain and serve to elucidate neural processes at the single-cell and circuit levels. Development of these tools is crucial for understanding complex behaviors and cognition and for advancing clinical applications. However, it remains a challenge to densely record from cell populations stably and continuously over long time periods. Many popular electrodes, such as tetrodes and silicon arrays, feature large cross-diameters that produce damage upon insertion and elicit chronic reactive tissue responses associated with neuronal death, hindering the recording of stable, continuous neural activity. In addition, most wire bundles exhibit broad spacing between channels, precluding simultaneous recording from a large number of cells clustered in a small area. The carbon fiber microelectrode arrays described in this protocol offer an accessible solution to these concerns. The study provides a detailed method for fabricating carbon fiber microelectrode arrays that can be used for both acute and chronic recordings in vivo. The physical properties of these electrodes make them ideal for stable and continuous long-term recordings at high cell densities, enabling the researcher to make robust, unambiguous recordings from single units across months.


Community structure of zooplankton in the main entrance of Bahía Magdalena, México during 1996.
The zooplankton community structure, including copepods, euphausiids, chaetognaths, and decapod larvae, was monitored during six circadian cycles using Bongo net (500 microns mesh net) samples from Bahía Magdalena, on the southwest coast of Baja California, México. Samples were obtained during three oceanographic surveys (March, July, and December 1996) to describe the changes in the zooplankton community structure throughout the main mouth of Bahía Magdalena. The zooplankton community structure showed strong changes with a close relation to environmental conditions. During March, a well-mixed water column with low temperature and salinity indicated an influence of the California Current water and local upwelling processes. During July, temperature increased and a wide salinity range was recorded. The stratification of the water column was intense during summer, enhancing the thermocline. The highest temperatures and salinity were recorded in December, related to the presence of the Costa Rica Coastal Current (CRCC). The thermocline deepened as water temperature increased. A typical temperate community structure with low specific richness dominated by Calanus pacificus, Nyctiphanes simplex, and Acartia clausi and high zooplankton biomass (average 9.3 and 5.5 ml 1000 m-3 respectively) during March and July shifted to a more complex tropical community structure with a low zooplankton biomass in December (average 0.37 ml 1000 m-3). The mouth of Bahía Magdalena has a vigorous exchange of water caused by tidal currents. The zooplankton community structure was not significantly different between the central part of Bahía Magdalena and the continental shelf outside the bay for all months. The results suggest a more dynamic inside-outside interaction of zooplankton assemblages than first thought.


Chronic Recurrent Multifocal Osteomyelitis Involving the Mandible - A Rare Case Report.
Chronic recurrent multifocal osteomyelitis (CRMO) is an uncommon, aseptic, autoinflammatory condition characterized by multifocal bone lesions with pain, swelling, and frequent exacerbations and remissions. It is noteworthy that these lesions occur without any identifiable etiology or microbiologic finding. The clavicle and metaphyses of the long bones are often involved whereas involvement of the mandible is considered rare. It is usually diagnosed by exclusion of other diseases. As it shares most of its features with the more commonly occurring infective osteomyelitis, patients are often unnecessarily subjected to prolonged courses of antibiotics, serial radiation exposures, and repeated bone biopsies. We present a case of CRMO involving the mandible. Our primary objective is to demonstrate the clinical features of this uncommon disorder, highlighting the radiographic appearance. Familiarity with this condition among radiologists greatly increases the likelihood for early diagnosis and formulating an appropriate treatment plan.


Adoptive cell therapy for patients with melanoma, using tumor-infiltrating lymphocytes genetically engineered to secrete interleukin-2.
Adoptive cell transfer of tumor-infiltrating lymphocytes (TILs) after lymphodepletion mediates regression in 50% of patients with metastatic melanoma. In vivo persistence and telomere length of the transferred cells correlate with antitumor response. In an attempt to prolong the in vivo survival of the transferred cells, TILs were genetically engineered to produce interleukin (IL)-2. In vitro, these transduced TILs secreted IL-2 while retaining tumor specificity and exhibited prolonged survival after IL-2 withdrawal. In a phase I/II clinical trial, seven evaluable patients received transduced TILs and one patient experienced a partial response associated with in vivo persistence of IL-2-transduced TILs in circulating lymphocytes. An additional five patients received transduced TILs in conjunction with IL-2 administration. Persistence of IL-2-transduced TILs was observed in three patients, including one partial responder. The transgene DNA as well as vector-derived IL2 mRNA could be detected for 4 months in responding patients. The low response rate in this trial was possibly due to a reduction in telomere length in cells as a result of prolonged in vitro culture. In this study, insertion of the IL-2 gene into antitumor TILs increased their ability to survive after IL-2 withdrawal in vitro but did not increase their in vivo persistence or clinical effectiveness.


Anaerovorax odorimutans gen. nov., sp. nov., a putrescine-fermenting, strictly anaerobic bacterium.
The strictly anaerobic, gram-positive, non-spore-forming bacterium strain NorPut1T ferments putrescine to acetate, butyrate, molecular hydrogen and ammonia. It also utilizes 4-aminobutyrate and 4-hydroxybutyrate as growth substrates. Comparative 16S rDNA sequence analysis confirmed a phylogenetic affiliation of this strain to the phylum of gram-positive bacteria with low DNA G+C content. Together with its closest relative, 'Clostridium aminobutyricum' (DSM 2634), and several Eubacterium species, strain NorPut1T represents a well-defined monophyletic group. Moderate overall 16S rRNA sequence similarities (< 91%) were found for the NorPut1T/'Clostridium aminobutyricum' pair and several Eubacterium species. The type species, Eubacterium limosum, is not a member of the group and, together with Eubacterium barkeri and Pseudoramibacter alactolyticus, represents a distant phylogentic cluster. Therefore, a new genus, Anaerovorax, is proposed as harbouring strain NorPut1T (= DSM 5092T), which is described as a new species, i.e. Anaerovorax odorimutans.


Attenuation of human immunodeficiency virus type 1 cytopathic effect by a mutation affecting the transmembrane envelope glycoprotein.
The cytopathic effects of human immunodeficiency virus type 1 (HIV-1) infection are specific for cells that express the CD4 viral receptor and consist of syncytium formation and single-cell lysis. Here we report that a mutation (517A) affecting the amino terminus of the HIV-1 gp41 transmembrane envelope glycoprotein resulted in a virus that was markedly less cytopathic than was wild-type HIV-1. In systems in which cell-to-cell transmission of HIV-1 occurred, the replication ability of the 517A virus was comparable with that of the wild-type virus. Even though the levels of viral protein expression, virion production, and interaction of the envelope glycoproteins with CD4 were similar for the 517A and wild-type viruses, both syncytium formation and single-cell lysis were attenuated for the 517A mutant virus. These results demonstrate that an envelope glycoprotein region important for mediating post-receptor binding events in cell membrane fusion is important for the induction of cytopathic effects by HIV-1. These results also indicate that levels of HIV-1 viral proteins or viral particles produced in infected cells are in themselves not sufficient to induce cytopathic effects.


Targeting of dicer-2 and RNA by a viral RNA silencing suppressor in Drosophila cells.
RNA interference (RNAi) is a eukaryotic gene-silencing mechanism that functions in antiviral immunity in diverse organisms. To combat RNAi-mediated immunity, viruses encode viral suppressors of RNA silencing (VSRs) that target RNA and protein components in the RNAi machinery. Although the endonuclease Dicer plays key roles in RNAi immunity, little is known about how VSRs target Dicer. Here, we show that the B2 protein from Wuhan nodavirus (WhNV), the counterpart of Flock House virus (FHV), suppresses Drosophila melanogaster RNAi by directly interacting with Dicer-2 (Dcr-2) and sequestering double-stranded RNA (dsRNA) and small interfering RNA (siRNA). Further investigations reveal that WhNV B2 binds to the RNase III and Piwi-Argonaut-Zwille (PAZ) domains of Dcr-2 via its C-terminal region, thereby blocking the activities of Dcr-2 in processing dsRNA and incorporating siRNA into the RNA-induced silencing complex (RISC). Moreover, we uncover an interrelationship among diverse activities of WhNV B2, showing that RNA binding enhances the B2-Dcr-2 interaction by promoting B2 homodimerization. Taken together, our findings establish a model of suppression of Drosophila RNAi by WhNV B2 targeting both Dcr-2 and RNA and provide evidence that an interrelationship exists among diverse activities of VSRs to antagonize RNAi.


Lateralizing language function with pre-operative functional magnetic resonance imaging in early proficient bilingual patients.
Research on bilinguals with brain lesions is complicated by high patient variability, making it difficult to find well-matched controls. We benefitted from a database of over 700 patients and conducted an analysis of pre-operative functional magnetic resonance imaging data to assess language dominance in 25 early, highly proficient Spanish-English bilinguals, and 25 carefully matched monolingual controls. Our results showed that early bilingualism is associated with greater bilateral hemispheric involvement, and monolingualism is associated with stronger left hemisphere lateralization (p=0.009). The bilinguals showed more pronounced right hemisphere activation (p=0.008). Although language dominance values were concordant in the bilingual group, there were a few (12%) atypical cases with different lateralization patterns in L1 and L2. Finally, we found distinct areas of activity in first and second language within the language network, in addition to regions of convergence. These data underscore the need to map all languages proficiently spoken by surgical candidates.


What Explains Socioeconomic Differences in the Speed of Heart Rate Recovery to Postural Challenge?
Much recent work has focused on the value of heart rate recovery (HRR) as a marker of cardiovascular health and a predictor of mortality. This article explores socioeconomic variation in HRR following exposure to a potent physiological stressor. The sample involved a nationally representative cohort of 4,475 community-dwelling older persons aged 50 years and older participating in the Irish Longitudinal Study on Ageing (TILDA). Participants completed an active stand (ie, vertical stand from a supine position) as part of a detailed clinic-based cardiovascular health assessment. Beat-to-beat HRR to standing was monitored over a 2-minute time horizon using a finometer. Highest level of educational achievement served as the indicator variable for socioeconomic status and mediation analysis was undertaken to explore the pathways through which social inequality comes to affect the speed of HRR using the extensive array of covariates available in TILDA. Participants with primary level education were characterized by a significantly slower HRR after standing compared with the tertiary educated (B = -1.15 bpm, CI95 = -1.78, -0.52; p < .001). Mediation analysis revealed that lifetime smoking accounted for a sizeable proportion (40.4%) of the educational differential. Adjustment for other objectively measured markers of lifestyle measured during the clinic visit accounted for only a small proportion (5.2%) of the difference. Smoking may represent a major pathway through which the social environment becomes biologically embedded in the tissues and organs of the body precipitating earlier vascular ageing among more socially disadvantaged groups, emphasizing the need to address the causes of these inequalities.


Optogenetic stimulation reveals a latent tipping point in cortical networks during ictogenesis.
Brain-state transitions are readily apparent from changes in brain rhythms,1 but are difficult to predict, suggestive that the underlying cause is latent to passive recording methods. Among the most important transitions, clinically, are the starts of seizures. We here show that an 'active probing' approach may have several important benefits for epileptic management, including by helping predict these transitions. We used mice expressing the optogenetic actuator, channelrhodopsin, in pyramidal cells, allowing this population to be stimulated in isolation. Intermittent stimulation at frequencies as low as 0.033 Hz (period = 30 s) delayed the onset of seizure-like events in an acute brain slice model of ictogenesis, but the effect was lost if stimulation was delivered at even lower frequencies (1/min). Notably, active probing additionally provides advance indication of when seizure-like activity is imminent, revealed by monitoring the postsynaptic response to stimulation. The postsynaptic response, recorded extracellularly, showed an all-or-nothing change in both amplitude and duration, a few hundred seconds before seizure-like activity began-a sufficient length of time to provide a helpful warning of an impending seizure. The change in the postsynaptic response then persisted for the remainder of the recording, indicative of a state change from a pre-epileptic to a pro-epileptic network. This occurred in parallel with a large increase in the stimulation-triggered Ca2+ entry into pyramidal dendrites, and a step increase in the number of evoked postsynaptic action potentials, both consistent with a reduction in the threshold for dendritic action potentials. In 0 Mg2+ bathing media, the reduced threshold was not associated with changes in glutamatergic synaptic function, nor of GABAergic release from either parvalbumin or somatostatin interneurons, but simulations indicate that the step change in the optogenetic response can instead arise from incremental increases in intracellular [Cl-]. The change in the response to stimulation was replicated by artificially raising intracellular [Cl-], using the optogenetic chloride pump, halorhodopsin. By contrast, increases in extracellular [K+] cannot account for the firing patterns in the response to stimulation, although this, and other cellular changes, may contribute to ictal initiation in other circumstances. We describe how these various cellular changes form a synergistic network of positive feedback mechanisms, which may explain the precipitous nature of seizure onset. This model of seizure initiation draws together several major lines of epilepsy research as well as providing an important proof-of-principle regarding the utility of open-loop brain stimulation for clinical management of the condition.


Iron deficiency.
Iron deficiency is one of the leading contributors to the global burden of disease, and particularly affects children, premenopausal women, and people in low-income and middle-income countries. Anaemia is one of many consequences of iron deficiency, and clinical and functional impairments can occur in the absence of anaemia. Iron deprivation from erythroblasts and other tissues occurs when total body stores of iron are low or when inflammation causes withholding of iron from the plasma, particularly through the action of hepcidin, the main regulator of systemic iron homoeostasis. Oral iron therapy is the first line of treatment in most cases. Hepcidin upregulation by oral iron supplementation limits the absorption efficiency of high-dose oral iron supplementation, and of oral iron during inflammation. Modern parenteral iron formulations have substantially altered iron treatment and enable rapid, safe total-dose iron replacement. An underlying cause should be sought in all patients presenting with iron deficiency: screening for coeliac disease should be considered routinely, and endoscopic investigation to exclude bleeding gastrointestinal lesions is warranted in men and postmenopausal women presenting with iron deficiency anaemia. Iron supplementation programmes in low-income countries comprise part of the solution to meeting WHO Global Nutrition Targets.


AACC Guidance Document on Laboratory Investigation of Acute Kidney Injury.
Acute kidney injury (AKI) is a sudden episode of kidney damage or failure affecting up to 15% of hospitalized patients and is associated with serious short- and long-term complications, mortality, and health care costs. Current practices to diagnose and stage AKI are variable and do not factor in our improved understanding of the biological and analytical variability of creatinine. In addition, the emergence of biomarkers, for example, cystatin C, insulin-like growth factor binding protein 7, and tissue inhibitor of metalloproteinases 2, and electronic notification tools for earlier detection of AKI, highlights the need for updated recommendations to address these developments. This AACC Academy guidance document is intended to provide laboratorians and clinicians up-to-date information regarding current best practices for the laboratory investigation of AKI. Topics covered include: clinical indications for further investigating potential AKI, analytical considerations for creatinine assays, the impact of biological variability on diagnostic thresholds, defining "baseline" creatinine, role of traditional markers (urine sodium, fractional excretion of sodium, fractional excretion of urea, and blood urea-to-creatinine ratio), urinary microscopic examination, new biomarkers, improving AKI-associated test utilization, and the utility of automated AKI alerts. The previous decade brought us a significant number of new studies characterizing the performance of existing and new biomarkers, as well as potential new tools for early detection and notification of AKI. This guidance document is intended to inform clinicians and laboratorians on the best practices for the laboratory investigation of AKI, based on expert recommendations where the preponderance of evidence is available.


BCG and protection against inflammatory and auto-immune diseases.
Bacillus Calmette-Guérin (BCG) is the only available vaccine against tuberculosis. Although its protective efficacy against pulmonary tuberculosis is still under debate, it provides protection against other mycobacterial diseases. BCG is also an effective therapy against superficial bladder cancer and potentially decreases overall childhood mortality. Areas covered: The purpose of this paper is to provide a state-of-the-art summary of the beneficial effects of BCG in inflammatory and auto-immune diseases. As a strong inducer of Th1 type immunity, BCG has been reported to protect against atopic conditions, such as allergic asthma, a Th2-driven disorder. Its protective effect has been well documented in mice, but still awaits definitive evidence in humans. Similarly, murine studies have shown a protective effect of BCG against auto-immune diseases, such as multiple sclerosis and insulin-dependent diabetes, but studies in humans have come to conflicting conclusions. Expert commentary: Studies in mice have shown a beneficial effect of the BCG vaccine against allergic asthma, multiple sclerosis and diabetes. However, the understanding of its mechanism is still fragmentary and requires further in depth research. Some observational or intervention studies in humans have also suggested a beneficial effect, but definitive evidence for this requires confirmation in carefully conducted prospective studies.


Quadratic Gabor filters for object detection.
We present a new class of quadratic filters that are capable of creating spherical, elliptical, hyperbolic and linear decision surfaces which result in better detection and classification capabilities than the linear decision surfaces obtained from correlation filters. Each filter comprises of a number of separately designed linear basis filters. These filters are linearly combined into several macro filters; the output from these macro filters are passed through a magnitude square operation and are then linearly combined using real weights to achieve the quadratic decision surface. For detection, the creation of macro filters (linear combinations of multiple single filters) allows for a substantial computational saving by reducing the number of correlation operations required. In this work, we consider the use of Gabor basis filters; the Gabor filter parameters are separately optimized. The fusion parameters to combine the Gabor filter outputs are optimized using an extended piecewise quadratic neural network (E-PQNN). We demonstrate methods for selecting the number of macro Gabor filters, the filter parameters and the linear and nonlinear combination coefficients. We present preliminary results obtained for an infrared (IR) vehicle detection problem.


Oxidative damage increases with degree of simulated bacterial infection, but not ectoparasitism, in tree swallow nestlings.
The purpose of mounting an immune response is to destroy pathogens, but this response comes at a physiological cost, including the generation of oxidative damage. However, many studies on the effects of immune challenges employ a single high dose of a simulated infection, meaning that the consequences of more mild immune challenges are poorly understood. We tested whether the degree of immunological challenge in tree swallows (Tachycineta bicolor) affects oxidative physiology and body mass, and whether these metrics correlate with parasitic nest mite load. We injected 14 day old nestlings with 0, 0.01, 0.1 or 1 mg lipopolysaccharide (LPS) per kg body mass, then collected a blood sample 24 h later to quantify multiple physiological metrics, including oxidative damage (i.e. d-ROMs), circulating amounts of triglyceride and glycerol, and levels of the acute phase protein haptoglobin. After birds had fledged, we identified and counted parasitic nest mites (Dermanyssus spp. and Ornithonyssus spp.). We found that only nestlings injected with 1 mg LPS kg-1 body mass, which is a common dosage in ecoimmunological studies, lost more body mass than individuals from other treatment groups. However, every dose of LPS resulted in a commensurate increase in oxidative damage. Parasitic mite abundance had no effect on oxidative damage across treatments. The amount of oxidative damage correlated with haptoglobin levels, suggesting compensatory mechanisms to limit self-damage during an immune response. We conclude that while only the highest-intensity immune challenges resulted in costs related to body mass, even low-intensity immune challenges result in detectable increases in oxidative damage.


Association of a MET genetic variant with autism-associated maternal autoantibodies to fetal brain proteins and cytokine expression.
The contribution of peripheral immunity to autism spectrum disorders (ASDs) risk is debated and poorly understood. Some mothers of children with ASD have autoantibodies that react to fetal brain proteins, raising the possibility that a subset of ASD cases may be associated with a maternal antibody response during gestation. The mechanism by which the maternal immune system breaks tolerance has not been addressed. We hypothesized that the mechanism may involve decreased expression of the MET receptor tyrosine kinase, an ASD risk gene that also serves as a key negative regulator of immune responsiveness. In a sample of 365 mothers, including 202 mothers of children with ASD, the functional MET promoter variant rs1858830 C allele was strongly associated with the presence of an ASD-specific 37+73-kDa band pattern of maternal autoantibodies to fetal brain proteins (P=0.003). To determine the mechanism of this genetic association, we measured MET protein and cytokine production in freshly prepared peripheral blood mononuclear cells from 76 mothers of ASD and typically developing children. The MET rs1858830 C allele was significantly associated with MET protein expression (P=0.025). Moreover, decreased expression of the regulatory cytokine IL-10 was associated with both the MET gene C allele (P=0.001) and reduced MET protein levels (P=0.002). These results indicate genetic distinction among mothers who produce ASD-associated antibodies to fetal brain proteins, and suggest a potential mechanism for how a genetically determined decrease in MET protein production may lead to a reduction in immune regulation.