H2A.Z controls the stability and mobility of nucleosomes to regulate expression of the LH genes.
The structure and dynamics of promoter chromatin have a profound effect on the expression levels of genes. Yet, the contribution of DNA sequence, histone post-translational modifications, histone variant usage and other factors in shaping the architecture of chromatin, and the mechanisms by which this architecture modulates expression of specific genes are not yet completely understood. Here we use optical tweezers to study the roles that DNA sequence and the histone variant H2A.Z have in shaping the chromatin landscape at the promoters of two model genes, Cga and Lhb. Guided by MNase mapping of the promoters of these genes, we reconstitute nucleosomes that mimic those located near the transcriptional start site and immediately downstream (+1), and measure the forces required to disrupt these nucleosomes, and their mobility along the DNA sequence. Our results indicate that these genes are basally regulated by two distinct strategies, making use of H2A.Z to modulate separate phases of transcription, and highlight how DNA sequence, alternative histone variants and remodelling machinery act synergistically to modulate gene expression.


Carboxy-Terminal Cementum Protein 1-Derived Peptide 4 (cemp1-p4) Promotes Mineralization through wnt/β-catenin Signaling in Human Oral Mucosa Stem Cells.
Human cementum protein 1 (CEMP1) is known to induce cementoblast and osteoblast differentiation and alkaline phosphatase (ALP) activity in human periodontal ligament-derived cells in vitro and promotes bone regeneration in vivo. CEMP1's secondary structure analysis shows that it has a random-coiled structure and is considered an Intrinsic Disordered Protein (IDP). CEMP1's short peptide sequences mimic the biological capabilities of CEMP1. However, the role and mechanisms of CEMP1's C-terminal-derived synthetic peptide (CEMP1-p4) in the canonical Wnt/β-catenin signaling pathway are yet to be described. Here we report that CEMP1-p4 promotes proliferation and differentiation of Human Oral Mucosa Stem Cells (HOMSCs) by activating the Wnt/β-catenin pathway. CEMP1-p4 stimulation upregulated the expression of β-catenin and glycogen synthase kinase 3 beta (GSK-3B) and activated the transcription factors TCF1/7 and Lymphoid Enhancer binding Factor 1 (LEF1) at the mRNA and protein levels. We found translocation of β-catenin to the nucleus in CEMP1-p4-treated cultures. The peptide also penetrates the cell membrane and aggregates around the cell nucleus. Analysis of CEMP1-p4 secondary structure revealed that it has a random-coiled structure. Its biological activities included the induction to nucleate hydroxyapatite crystals. In CEMP1-p4-treated HOMSCs, ALP activity and calcium deposits increased. Expression of Osterix (OSX), Runt-related transcription factor 2 (RUNX2), Integrin binding sialoproptein (IBSP) and osteocalcin (OCN) were upregulated. Altogether, these data show that CEMP1-p4 plays a direct role in the differentiation of HOMSCs to a "mineralizing-like" phenotype by activating the β-catenin signaling cascade.


Multiple determinants and consequences of cohesion fatigue in mammalian cells.
Cells delayed in metaphase with intact mitotic spindles undergo cohesion fatigue, where sister chromatids separate asynchronously, while cells remain in mitosis. Cohesion fatigue requires release of sister chromatid cohesion. However, the pathways that breach sister chromatid cohesion during cohesion fatigue remain unknown. Using moderate-salt buffers to remove loosely bound chromatin cohesin, we show that "cohesive" cohesin is not released during chromatid separation during cohesion fatigue. Using a regulated protein heterodimerization system to lock different cohesin ring interfaces at specific times in mitosis, we show that the Wapl-mediated pathway of cohesin release is not required for cohesion fatigue. By manipulating microtubule stability and cohesin complex integrity in cell lines with varying sensitivity to cohesion fatigue, we show that rates of cohesion fatigue reflect a dynamic balance between spindle pulling forces and resistance to separation by interchromatid cohesion. Finally, while massive separation of chromatids in cohesion fatigue likely produces inviable cell progeny, we find that short metaphase delays, leading to partial chromatid separation, predispose cells to chromosome missegregation. Thus, complete separation of one or a few chromosomes and/or partial separation of sister chromatids may be an unrecognized but common source of chromosome instability that perpetuates the evolution of malignant cells in cancer.


Determination of cytomegalovirus antibodies by enzyme-linked immunosorbent assay (ELISA) in women of fertile period.
Such cytomegalovirus antibodies as the IgG and IgM are investigated in the present work. Investigation was carried out on 301 sera of fertile aged women (between 18 to 35 years of age) under the enzyme-linked immunosorbent assay (ELISA). Only one dilution has been made i.e. 1:40 and the final dilution was worked out from regression graph obtained from a previously titred human positive serum analysis; IgG antibodies were found in 265 cases (88%), appearing IgM antibodies in 4 of them, with the remaining 36 cases showing neither IgG nor IgM antibodies (12%). These results were compared with those obtained in other countries and the importance of the serological diagnosis to this virus was pointed out.


ABCF2, an Nrf2 target gene, contributes to cisplatin resistance in ovarian cancer cells.
Previously, we have demonstrated that NRF2 plays a key role in mediating cisplatin resistance in ovarian cancer. To further explore the mechanism underlying NRF2-dependent cisplatin resistance, we stably overexpressed or knocked down NRF2 in parental and cisplatin-resistant human ovarian cancer cells, respectively. These two pairs of stable cell lines were then subjected to microarray analysis, where we identified 18 putative NRF2 target genes. Among these genes, ABCF2, a cytosolic member of the ABC superfamily of transporters, has previously been reported to contribute to chemoresistance in clear cell ovarian cancer. A detailed analysis on ABCF2 revealed a functional antioxidant response element (ARE) in its promoter region, establishing ABCF2 as an NRF2 target gene. Next, we investigated the contribution of ABCF2 in NRF2-mediated cisplatin resistance using our stable ovarian cancer cell lines. The NRF2-overexpressing cell line, containing high levels of ABCF2, was more resistant to cisplatin-induced apoptosis compared to its control cell line; whereas the NRF2 knockdown cell line with low levels of ABCF2, was more sensitive to cisplatin treatment than its control cell line. Furthermore, transient overexpression of ABCF2 in the parental cells decreased apoptosis and increased cell viability following cisplatin treatment. Conversely, knockdown of ABCF2 using specific siRNA notably increased apoptosis and decreased cell viability in cisplatin-resistant cells treated with cisplatin. This data indicate that the novel NRF2 target gene, ABCF2, plays a critical role in cisplatin resistance in ovarian cancer, and that targeting ABCF2 may be a new strategy to improve chemotherapeutic efficiency.


Cervical Radiculopathy: Incidence and Treatment of 1,420 Consecutive Cases.
Retrospective case series. To determine the incidence of cervical radiculopathy requiring operative intervention by level and to report on the methods of treatment. Cervical radiculopathy is a common cause of pain and can result in progressive neurological deficits. Although the pathology is well understood, the actual incidence of cervical radiculopathy at particular spinal levels ultimately requiring operative intervention is unknown. A large consecutive series of patients operated on by a single surgeon were retrospectively analyzed. The incidence of cervical radiculopathy at each level was defined for every patient. Procedures used for operative treatment were noted. Health related quality of life (HRQL) scores were collected both pre-operatively and postoperatively. There were 1305 primary and 115 revision operations performed. The most common primary procedures performed were anterior cervical discectomy and fusion (ACDF, 50%) and anterior cervical corpectomy and fusion (ACCF, 28%). The most commonly affected levels were C6 (66%) and C7 (62%). Reasons for revision were pseudarthrosis (27%), clinical adjacent segment pathology (CASP, 63%), persistent radiculopathy (11%), and hardware-related (2.6%). The most common procedures performed in the revision group were posterior cervical decompression and fusion (PCDF, 42%) and ACDF (40%). The most commonly affected levels were C7 (43%) and C5 (30%). Among patients that had their index surgery at our institution, the revision rate was 6.4%. In both primary and revision cases there was a significant improvement in Neck Disability Index and visual analogue scale scores postoperatively. Postoperative HRQL scores in the revision cases were significantly worse than those in the primary cases (p <0.01). This study provides the largest description of the incidence of cervical radiculopathy by level and operative outcomes in patients undergoing cervical decompression. The incidence of CASP was 4.2% in 3.3 years in this single institution series.


The effect of dioxyphenylalanine (DOPA), amides and some potential sources of energy on the multiplication of Mycobacterium leprae.
The multiplication of 2 out of 3 M. leprae strains on a medium containing substances from digested non-acid-fast microorganisms or even free of them was significally promoted to D-3,4-dihydroxyphenylalanine (DOPA). The following organic substances exerted growth-promoting effects on several strains: 0.02-0.10% concentrations of succinate greater than fumarate greater than alpha-ketoglutarate and acetate greater than glycerol; 0.2% concentrations of citrate and pyruvate greater than isonicotinamide and benzamide greater than lecithin. 0.5% concentrations of oleate greater than citrate greater than pyruvate greater than acetate greater than fumarate greater than succinate; 5.0% concentrations of butanol and butandiol greater than propanol greater than sorbitol greater than ethanol. However, these effects were variable and strains of various origins acted differently. On media containing DOPA, malachite-green (MG) and at least 0.12 x 10(6) microorganisms/ml the following oxidation-reduction reaction was observed: DOPA was oxidized to a brown compound and malachite-green reduced to an almost colourless product. Consequently, the blue-colour of the cultures turned from blue to brown. This DOPA-MG reaction and the inability to grow on conventional media were used for the identification of 10 cultures (inocula directly from patients) as M. leprae, while a DOPA-MG negative patient-strain grew on media employed for cultivable mycobacteria and was not identical with M. leprae, as proved by the foot-pad test in mice.


Transgene-encoded human CD2 acts in a dominant negative fashion to modify thymocyte selection signals in mice.
CD2 is a cell surface glycoprotein present on all T cells which has been shown to function as an adhesion and signaling molecule. Expressed early in T cell development, human CD2 (HCD2) has been suggested to play a role during thymopoiesis. However, the relevance of CD2 in T cell development has been called into question recently, as neither disruption of the CD2 gene nor anti-CD2 antibody treatment of fetal thymic organ cultures in mouse were shown to have any discernible consequences. We have expressed HCD2 at high levels in transgenic mice and found a profound effect of the transgene on thymocyte differentiation. Transgenic thymuses are considerably reduced in cell number as a consequence of increased apoptosis of double-positive (DP) thymocytes in the cortex. The remaining DP cells have up-regulated levels of T cell receptor (TCR) and are resistant to apoptosis mediated by administration of antigen. These effects are dependent on the cytoplasmic domain of HCD2, as mice expressing comparable levels of a tailless HCD2 transgene have a normal phenotype. The HCD2 cytoplasmic domain contains several regions of identity with mouse CD2 and can interact effciently with mouse intracellular signaling machinery. These results suggest there is considerable cross-talk between CD2 and TCR on developing thymocytes with consequences for the stimulation threshold of mature T cells.


Protein compactness and interaction valency define the architecture of a biomolecular condensate across scales.
Non-membrane-bound biomolecular condensates have been proposed to represent an important mode of subcellular organization in diverse biological settings. However, the fundamental principles governing the spatial organization and dynamics of condensates at the atomistic level remain unclear. The Saccharomyces cerevisiae Lge1 protein is required for histone H2B ubiquitination and its N-terminal intrinsically disordered fragment (Lge11-80) undergoes robust phase separation. This study connects single- and multi-chain all-atom molecular dynamics simulations of Lge11-80 with the in vitro behavior of Lge11-80 condensates. Analysis of modeled protein-protein interactions elucidates the key determinants of Lge11-80 condensate formation and links configurational entropy, valency, and compactness of proteins inside the condensates. A newly derived analytical formalism, related to colloid fractal cluster formation, describes condensate architecture across length scales as a function of protein valency and compactness. In particular, the formalism provides an atomistically resolved model of Lge11-80 condensates on the scale of hundreds of nanometers starting from individual protein conformers captured in simulations. The simulation-derived fractal dimensions of condensates of Lge11-80 and its mutants agree with their in vitro morphologies. The presented framework enables a multiscale description of biomolecular condensates and embeds their study in a wider context of colloid self-organization.


Functional and Structural Brain Correlates of Socioeconomic Status.
Socioeconomic status (SES) is a multidimensional construct that includes not only measures of material wealth, but also education, social prestige, and neighborhood quality. Socioeconomic correlates between wealth and cognitive functions have been well established in behavioral studies. However, functional and structural brain correlates of SES remain unclear. Here, we sought to uncover the most likely neural regions to be affected by low SES, specifically associated with age. Using effect size-seed-based d Mapping, we compiled studies that examined individuals with low SES and performed functional magnetic resonance imaging and voxel-based morphometry meta-analyses. The results revealed that as from early to late age, individuals exposed to low SES are less likely to have sustained executive network activity yet a greater likelihood to enhanced activity within reward-related regions. A similar activity was shown for gray matter volume across early to older age. These findings provide the first quantitative integration of neuroimaging results pertaining to the neural basis of SES. Hypoactivation of the executive network and hyperactivation of the reward network in low SES individuals may support the scarcity hypothesis and animal models of the effects of early adversity.


Gene expression profiling of MicroRNAs in small-bowel transplantation paraffin-embedded mucosal biopsy tissue.
The molecular mechanisms and regulation of immune-mediated rejection of organ allografts remains unclear. Recent studies have reported that small non-coding RNAs, microRNAs (miRNAs) play a critical role in the immune system via modulation of transcription and translation. We hypothesized that particular miRNAs provide regulation of an ensuing intragraft immune effector response. The aim of our study was to detect miRNAs involved in acute cellular rejection (AR) in human small intestinal allografts. We examined 12 small intestinal mucosal biopsies (AR, 7 cases, all grade 2 or 3) and non-rejecting (NR) allografts (5 cases, all grade 0) obtained from recipients after small bowel or multivisceral transplantation. RNA was isolated from the formalin-fixed paraffin-embedded (FFPE) biopsy samples and transcribed to cDNA. After preamplification we utilized a PCR based TaqMan Low Density Array (TLDA) containing 365 mature human miRNAs. Relative quantification was done based on pooled normal intestine using a comparative Ct method. We identified 62 miRNA upregulated genes in small bowels with ACR, and 35 were downregulated. Forty-two miRNA genes were upregulated in non-ACR small bowel biopsy samples (grade IND), and 45 were downregulated. The relative fold change ratio of ACR to non-ACR was calculated, and 50 upregulated and 8 downregulated miRNAs were detected as significant. Several interesting miRNAs will be evaluated further from this preliminary study. Our data suggests that intragraft miRNAs are potentially involved in the activation of a host alloimmune response to donor. These miRNAs may serve as targets for appropriate intervention and may be useful to monitor the allograft status.


Apparent radii of the native, stable intermediates and unfolded conformers of the alpha-subunit of tryptophan synthase from E. coli, a TIM barrel protein.
The urea-induced equilibrium unfolding of the alpha-subunit of tryptophan synthase (alphaTS) from Escherichia coli can be described by a four-state model, N right harpoon over left harpoon I1 right harpoon over left harpoon I2 right harpoon over left harpoon U, involving two highly populated intermediates, I1 and I2 [Gualfetti, P. J., Bilsel, O., and Matthews, C. R. (1999) Protein Sci. 8, 1623-1635]. To extend the physical characterization of these stable forms, the apparent radius was measured by several techniques. Size-exclusion chromatography (SEC), analytical ultracentrifugation (UC), and dynamic light scattering (DLS) experiments yield an apparent Stokes radius, R(s), of approximately 24 A for the native state of alphaTS. The small-angle X-ray scattering (SAXS) experiment yields a radius of gyration, R(g), of 19.1 A, consistent with the value predicted from the X-ray structure and the Stokes radius. As the equilibrium is shifted to favor I1 at approximately 3.2 M and I2 at 5.0 M urea, SEC and UC show that R(s) increases from approximately 38 to approximately 52 A. Measurements of the radius by DLS and SAXS between 2 and 4.5 M urea were complicated by the self-association of the I1 species at the relatively high concentrations required by those techniques. Above 6 M urea, SEC and UC reveal that R(s) increases linearly with increasing urea concentration to approximately 54 A at 8 M urea. The measurements of R(s) by DLS and R(g) by SAXS are sufficiently imprecise that both values appear to be identical for the I2 and U states and, considering the errors, are in good agreement with the results from SEC and UC. Thermodynamic parameters extracted from the SEC data for the N right harpoon over left harpoon I1 and I1 right harpoon over left harpoon I2 transitions agree with those from the optical data, showing that this technique accurately monitors a part of the equilibrium model. The lack of sensitivity to the I2 right harpoon over left harpoon U transition, beyond a simple swelling of both species with increasing urea concentration, implies that the Stokes radii for the I2 and U states are not distinguishable. Surprisingly, the hydrophobic core known to stabilize I2 at 5.0 M urea [Saab-Rincón, G., Gualfetti, P. J., and Matthews, C. R. (1996) Biochemistry 35, 1988-1994] develops without a significant contraction of the polypeptide, i.e., beyond that experienced by the unfolded form at decreasing urea concentrations. Kratky plots of the SAXS data, however, reveal that I2, similar to N and I1, has a globular structure while U has a more random coil-like form. By contrast, the formation of substantial secondary structure and the burial of aromatic side chains in I1 and, eventually, N are accompanied by substantial decreases in their Stokes radii and, presumably, the size of their respective conformational ensembles.


Pass-through of the Oakland, California, sugar-sweetened beverage tax in food stores two years post-implementation: A difference-in-differences study.
Taxes on sugar-sweetened beverages (SSBs) have gained support as a policy response to adverse health effects associated with SSB consumption. On July 1, 2017, Oakland, California, implemented a one-cent/ounce tax on SSBs with ≥25 calories/12 fluid ounces. This study estimated the long-term impact of the tax on taxed and untaxed beverage prices. Data on 5,830 taxed and 5,146 untaxed beverage prices were obtained from 99 stores in Oakland and 111 stores in Sacramento (comparison site), California, in late May-June 2017 and June 2019. Linear regression difference-in-differences models were computed with store and product fixed effects, with robust standard errors clustered on store, weighted based on volume sold by beverage sweetener status, type, and size. Taxed beverage prices increased by 0.73 cents/ounce (95% CI = 0.47,1.00) on average in supermarkets and grocery stores in Oakland relative to Sacramento and 0.74 cents/ounce (95% CI = 0.39,1.09) in pharmacies, but did not change in convenience stores (-0.09 cents/ounce, 95% CI = -0.56,0.39). Untaxed beverage prices overall increased by 0.40 cents/ounce (95% CI = 0.05,0.75) in pharmacies but did not change in other store types. Prices of taxed individual-size soda specifically increased in all store types, by 0.91-2.39 cents/ounce (p<0.05), as did prices of untaxed individual-size soda in convenience stores (0.79 cents/ounce, 95% CI = 0.01,1.56) and pharmacies (1.66 cents/ounce, 95% CI = 0.09,3.23). Two years following SSB tax implementation, there was partial tax pass-through with differences by store type and by beverage type and size within store type.


Elevated serum levels of arachidonoyl-lysophosphatidic acid and sphingosine 1-phosphate in systemic sclerosis.
Systemic sclerosis (SSc) is an often fatal disease characterized by autoimmunity and inflammation, leading to widespread vasculopathy and fibrosis. Lysophosphatidic acid (LPA), a bioactive phospholipid in serum, is generated from lysophospholipids secreted from activated platelets in part by the action of lysophospholipase D (lysoPLD). Sphingosine 1-phosphate (S1P), a member of the bioactive lysophospholipid family, is also released from activated platelets. Because activated platelets are a hallmark of SSc, we wanted to determine whether subjects with SSc have altered serum lysophospholipid levels or lysoPLD activity. Lysophospholipid levels were measured using mass spectrometric analysis. LysoPLD activity was determined by quantifying choline released from exogenous lysophosphatidylcholine (LPC). The major results were that serum levels of arachidonoyl (20:4)-LPA and S1P were significantly higher in SSc subjects versus controls. Furthermore, serum LPA:LPC ratios of two different polyunsaturated phospholipid molecular species, and also the ratio of all species combined, were significantly higher in SSc subjects versus controls. No significant differences were found between other lysophospholipid levels or lysoPLD activities. Elevated 20:4 LPA, S1P levels and polyunsaturated LPA:LPC ratios may be markers for and/or play a significant role in the etiology of SSc and may be future pharmacological targets for SSc treatment.


Histological Features of Flagellate Erythema.
Flagellate erythema is a distinctive eruption characterized by "whip-like" linear or curvilinear streaks and plaques, occurring mainly on the trunk. It has classically been described in 2 disparate clinical settings: chemotherapy with bleomycin and ingestion of mushrooms (most commonly Shiitake mushrooms). Most of the literature comprises single case reports, often with minimal histological description of rather nonspecific features. We describe in detail the histological features of 3 cases of flagellate erythema (2 related to bleomycin therapy and one related to ingestion of mushrooms) and review the findings described in the literature to define the spectrum of histological changes encountered in this eruption. Our 3 cases showed mild epidermal changes, with spongiosis and variable interface inflammation. All 3 showed a relatively prominent dermal lymphohistiocytic infiltrate, with features suggestive of a lymphocytic vasculopathy extending to at least the mid-reticular dermis. Eosinophils were a prominent component of the inflammatory infiltrate in 2 cases. Our review of the literature identified a total of 45 publications, representing reports of 46 patients, containing histological information. As well as bleomycin- and mushroom-related cases, similar eruptions have been reported in the context of connective tissue disease and other drugs. Although cases related to connective tissue disease show features of the underlying condition, cases secondary to drugs or mushrooms predominantly show features compatible with common patterns of exanthematous/morbilliform drug reaction. In particular, subtle spongiosis and/or interface dermatitis combined with a dermal lymphocytic infiltrate that includes increased numbers of eosinophils is a common finding. Features of a lymphocytic vasculopathy may be seen in a subset of these cases.


Long-term outcomes of an abstinence-based, small-group pregnancy prevention program in New York City schools.
Despite drops in U.S. teenage birthrates, questions continue to arise about how best to reduce the country's adolescent birthrate. School-based programs continue to be considered one of the best ways to reach adolescents at risk of early sexual activity. A total of 312 students completed a pretest, a posttest and a follow-up one year after the posttest: 125 who had participated in a 3-4-month-long abstinence-based small-group intervention led by trained social workers, and 187 in a comparison group that received no special services. There were few significant differences between the intervention and comparison groups at posttest. At the one-year follow-up, however, intervention students had significantly better scores on locus of control, their relationship with their parents and (among males only) their attitudes about the appropriateness of teenage sex. Measures of depression, self-esteem, intentions to have sex, attitudes toward teenage pregnancy and various behaviors did not differ significantly between groups. By the time of the one-year follow-up, there was no difference between study groups among females in the initiation of sexual intercourse. Among the males, initiation of sexual intercourse appeared to be higher in the intervention group than in the comparison group, but the difference was not statistically significant. Positive outcomes were especially limited among students who were already sexually active at the start of the study, a finding that emphasizes the difficulties of reaching adolescents who are already at high risk for pregnancy A small-group abstinence-based intervention focusing on mental health can have some impact on adolescents' attitudes and relationships (particularly with their parents). Long-term evaluations are important for determining the effects of an intervention, as it is difficult to change adolescent risk behavior.


Surface complexation modeling of Cu(II) adsorption on mixtures of hydrous ferric oxide and kaolinite.
The application of surface complexation models (SCMs) to natural sediments and soils is hindered by a lack of consistent models and data for large suites of metals and minerals of interest. Furthermore, the surface complexation approach has mostly been developed and tested for single solid systems. Few studies have extended the SCM approach to systems containing multiple solids. Cu adsorption was measured on pure hydrous ferric oxide (HFO), pure kaolinite (from two sources) and in systems containing mixtures of HFO and kaolinite over a wide range of pH, ionic strength, sorbate/sorbent ratios and, for the mixed solid systems, using a range of kaolinite/HFO ratios. Cu adsorption data measured for the HFO and kaolinite systems was used to derive diffuse layer surface complexation models (DLMs) describing Cu adsorption. Cu adsorption on HFO is reasonably well described using a 1-site or 2-site DLM. Adsorption of Cu on kaolinite could be described using a simple 1-site DLM with formation of a monodentate Cu complex on a variable charge surface site. However, for consistency with models derived for weaker sorbing cations, a 2-site DLM with a variable charge and a permanent charge site was also developed. Component additivity predictions of speciation in mixed mineral systems based on DLM parameters derived for the pure mineral systems were in good agreement with measured data. Discrepancies between the model predictions and measured data were similar to those observed for the calibrated pure mineral systems. The results suggest that quantifying specific interactions between HFO and kaolinite in speciation models may not be necessary. However, before the component additivity approach can be applied to natural sediments and soils, the effects of aging must be further studied and methods must be developed to estimate reactive surface areas of solid constituents in natural samples.


Predicting relapse in schizophrenia: Is BDNF a plausible biological marker?
Understanding the biological processes that underlie why patients relapse is an issue of fundamental importance to the detection and prevention of relapse in schizophrenia. Brain Derived Neurotrophic Factor (BDNF), a facilitator of brain plasticity, is reduced in patients with schizophrenia. In the present study, we examined whether decreases in plasma BDNF levels could be used as a biological predictor of relapse in schizophrenia. A total of 221 patients were prospectively evaluated for relapse over 30months in the Preventing Relapse in Schizophrenia: Oral Antipsychotics Compared to Injectables: eValuating Efficacy (PROACTIVE) study. Serial blood samples were collected at a maximum of 23 time points during the 30-month trial and BDNF levels were measured in plasma samples by ELISA. Receiver Operating Characteristic (ROC) curve analysis indicated that BDNF was not a significant predictor of relapse, hospitalization or exacerbation. Regardless of treatment group (oral second generation antipsychotic vs. long-acting injectable risperidone microspheres), baseline BDNF value did not differ significantly between those who experienced any of the adverse outcomes and those who did not. While contrary to the study hypothesis, these robust results offer little support for the use of plasma BDNF alone as a biomarker to predict relapse in schizophrenia.


Virological Control by the CD4-Binding Site Antibody N6 in Simian-Human Immunodeficiency Virus-Infected Rhesus Monkeys.
Passive immunotherapy against HIV-1 will most likely require broadly neutralizing antibodies (bnAbs) with maximum breadth and potency to ensure therapeutic efficacy. Recently, the novel CD4 binding site antibody N6 demonstrated extraordinary neutralization breadth and potency against large panels of cross-clade pseudoviruses. We evaluated the in vivo antiviral activity of N6-LS, alone or in combination with the established V3-glycan antibody PGT121, in chronically simian-human immunodeficiency virus (SHIV)-SF162P3-infected macaques. A single dose of N6-LS suppressed plasma viral loads in 4 out of 5 animals at day 7, while the combination of both antibodies suppressed all animals. The combination of both antibodies had no additive antiviral effect compared to a single dose of PGT121, potentially reflecting the nearly 10-fold-higher potency of PGT121 against this SHIV. Viral rebound occurred in the majority of suppressed animals and was linked to declining plasma bnAb levels over time. In addition to the effect on plasma viremia, bnAb administration resulted in significantly reduced proviral DNA levels in PBMCs after 2 weeks and in lymph nodes after 10 weeks. Autologous neutralizing antibody (nAb) responses and CD8+ T-cell responses were not significantly enhanced in the bnAb-treated animals compared to control animals, arguing against their contribution to the viral effects observed. These results confirm the robust antiviral activity of N6-LS in vivo, supporting the further clinical development of this antibody.IMPORTANCE Monocloncal antibodies (MAbs) are being considered for passive immunotherapy of HIV-1 infection. A critical requirement for such strategies is the identification of MAbs that recognize the diversity of variants within circulating but also reservoir viruses, and MAb combinations might be needed to achieve this goal. This study evaluates the novel bnAb N6-LS alone or in combination with the bnAb PGT121, in rhesus macaques that were chronically infected with SHIV. The results demonstrate that N6-LS potently suppressed plasma viral loads in the majority of animals but that the combination with PGT121 was not superior to PGT121 alone in delaying time to viral rebound or reducing peripheral blood mononuclear cell (PBMC) or lymph node proviral DNA levels. The occurrence of viral escape variants in an N6-LS-monotreated animal, however, argues for the need to maximize breadth and antiviral efficacy by combining bnAbs for therapeutic indications.


The environmental neurotoxin β-N-methylamino-l-alanine (l-BMAA) is deposited into birds' eggs.
The neurotoxic amino acid β-N-methylamino-L-alanine (BMAA) has been implicated in the etiology of neurodegenerative disorders. BMAA is also a known developmental neurotoxin and research indicates that the sources of human and wildlife exposure may be more diverse than previously anticipated. The aim of the present study was therefore to examine whether BMAA can be transferred into birds' eggs. Egg laying quail were dosed with 14C-labeled BMAA. The distribution of radioactivity in the birds and their laid eggs was then examined at different time points by autoradiography and phosphoimaging analysis. To evaluate the metabolic stability of the BMAA molecule, the distribution of 14C-methyl- and 14C-carboxyl-labeled BMAA were compared. The results revealed a pronounced incorporation of radioactivity in the eggs, predominantly in the yolk but also in the albumen. Imaging analysis showed that the concentrations of radioactivity in the liver decreased about seven times between the 24h and the 72h time points, while the concentrations in egg yolk remained largely unchanged. At 72h the egg yolk contained about five times the concentration of radioactivity in the liver. Both BMAA preparations gave rise to similar distribution pattern in the bird tissues and in the eggs, indicating metabolic stability of the labeled groups. The demonstrated deposition into eggs warrants studies of BMAAs effects on bird development. Moreover, birds' eggs may be a source of human BMAA exposure, provided that the laying birds are exposed to BMAA via their diet.


Expression of individual lamins in basal cell carcinomas of the skin.
In this study we used a unique collection of type specific anti-lamin antibodies to study lamin expression patterns in normal human skin and in skin derived from patients with basal cell carcinomas (BCCs). Lamin expression in serial sections from frozen tissue samples was investigated by single and double indirect immunofluorescence. In normal skin, lamin A was expressed in dermal fibroblasts and in suprabasal epithelial cells but was absent from all basal epithelial cells. Lamin C was expressed in dermal fibroblasts, suprabasal epithelial cells and a majority of basal epithelial cells. However, lamin C was not expressed in quiescent basal epithelial cells. Lamin B1 was expressed in all epithelial cells but was not expressed in dermal fibroblasts. Finally, lamin B2 was expressed in all epithelial cells but was not expressed in dermal fibroblasts. Finally, lamin B2 was expressed in all cell types in normal skin. Lamin expression was also investigated in a collection of 16 BCCs taken from a variety of body sites. Based upon patterns of lamin expression the BCCs were classified into four groups: A-negative (10/16 tumours), C-negative (5/16 tumours), A/C-negative (1/16 tumours) and A/B2-negative (1/16 tumours). Lamin expression was also compared to cell proliferation index by staining serial sections with the proliferation marker Ki67. 9/10 of the lamin A negative tumours were highly proliferative, whereas 4/5 of the lamin C negative tumours were slow growing. Thus as a general rule absence of lamin A was correlated with rapid growth within the tumour, while absence of lamin C was correlated with slow growth within the tumour. Our data supports the hypothesis that lamin A has a negative influence on cell proliferation and its down regulation may be a requisite of tumour progression.


Enhanced pressor response to centrally administered vasopressin in WKY rats on high sodium diet.
Four week old Wistar-Kyoto (WKY) rats were divided into two groups. The experimental group (n = 7) was receiving a high sodium diet (3.28% Na) and the control group (n = 7) a normal sodium diet (0.22% Na). After 8 weeks, subjects were chronically implanted with the lateral cerebral ventricle (LCV) cannulas and with the femoral artery catheters. Three series of experiments were carried out on the experimental and control groups. In each series mean arterial pressure (MAP) and heart rate (HR) were recorded for 10 min before and 30 min after the LCV infusion. In series 1 artificial cerebrospinal fluid (aCSF) was administered (2 microl/15 s). In series 2 AVP was infused (20 ng/2 microl aCSF/15 s). In series 3 V1a receptor antagonist (V1 ANT), d(CH2)5[Tyr(Me)2,Ala-NH29]AVP, was applied (80 ng/microl aCSF/15 s). There was no difference in baseline MAP and HR between the experimental and control groups. LCV infusion of aCSF had no effect on MAP and HR. LCV infusion of AVP produced a significant increase of MAP, which was greater in the group on the high sodium diet than in the group on normal sodium diet. The experimental group showed a longer hypertensive effect of centrally applied AVP in comparison to the control. LCV administration of V1 ANT did not exert a significant effect on circulatory parameters. These results suggest that the prolonged high sodium diet does not induce hypertension in WKY rats, but it enhances the pressor function of the central vasopressinergic system.


Evaluation of factors affecting bonding rate of calcium phosphate ceramic coatings for in vivo strain gauge attachment.
The aim of this study was to compare the bone-bonding rates of eight calcium phosphate ceramic (CPC) coatings attached to strain gauges, alone and in conjunction with an OP1 device (Creative BioMolecules, Hopkinton, MA) and autologous concentrated pericyte cells. These coatings were studied to develop faster bone bonding to long-term in vivo strain sensors. Characterization of the CPC powders using electron microscopy and X-ray diffraction showed that they had shapes ranging from spherical to rocklike and properties ranging from highly crystalline to amorphous. CPC coated gauges were placed on the femora of young male dogs during aseptic surgery and were initially held in place using resorbable sutures. Test groups were euthanized after 3, 9, and 12 weeks. Both femora of the dogs were explanted and cantilever loaded. Response of the implanted hydroxyapatite (HA) coated gauges were compared to the response of bench-top glued sets of gauges (controls) attached to the contralateral femur and reported as a percentage of the control values. One CPC coating type showed an average response of 30% of controls after 3 weeks, four showed average responses higher than 75% after 9 weeks, and three showed averages higher than 82% after 12 weeks in vivo. Amorphous CPC coatings bonded more quickly than crystalline ones and particle shape had less effect than crystal structure on bonding rates. When either OP1 or autologous concentrated pericyte cells were placed on selected CPC coated gauge surfaces, the CPC5 coated gauges bonded best after 3 weeks with a response of 59%. After the same time period in vivo, CPC3 and CPC7 provided responses of 40 and 16%, respectively. Comparison of a soluble calcium-coated CPC with an uncoated one that had identical crystal structure and similar particle shape indicated that the calcium coating slowed bone bonding substantially in the young dog model. Optical microscopy of stained undecalcified bone sections and backscattered electron imaging indicated bone formation at all bone-HA interfaces and an increase in the number of areas of bone remodeling adjacent to the gauge at all time periods. Gross bone remodeling due to strain gauge placement was only observed near the distalmost cell-seeded strain gauges. Selection of the type of coating and enhancement system can accelerate bone bonding to strain sensors but must be tailored to the bone of the model in which it is being used. Augmentation of CPC coatings with cells or OP1 resulted in variable enhancement of the bonding rate and depended on the CPC and the enhancement system.


Prevention of sternal wound infection in pediatric cardiac surgery: a protocolized approach.
Sternal wound infections (SWIs) are a costly complication for children after cardiac surgery, increasing morbidity, mortality, and financial cost. There are no pediatric guidelines to reduce the incidence of SWI in this vulnerable population. A quality improvement, multidisciplinary team was formed, and a protocol to prevent SWI was developed. A prospective review of patients who underwent pediatric cardiac surgery was conducted over a two-year period to follow adherence to the protocol and incidence of SWI. The Centers for Disease Control definitions for surgical site infections were used to determine the depth and presence of infection. Three hundred and eight children <18 years of age had sternotomies during the study period. There was a reduction in all SWI between the first and second years of the study (odds ratio [OR] = 0.35; confidence interval [CI] 95% 0.12-1.01; P = .059). Delayed sternal closure (DSC) was associated with increased risk of SWI (OR = 5.4; CI 95% 2.13-14.9; P ≤ .001). Institution of a protocol in patients with DSC was associated with decreased infections during the second year (first year: n = 7 (14%), second year: n = 2 (4%), P = .14). Institution of a protocol was associated with a decreased number of infections in children. A multicenter study of a bundled protocol approach to SWI prevention is needed. Children with DSC had a significantly higher risk of developing a wound infection. Initiating strategies to reduce SWI with a focus on children with DSC may result in improved overall infection rates.


Precipitation effects on microbial pollution in a river: lag structures and seasonal effect modification.
The river Göta Älv is a source of freshwater for 0.7 million swedes. The river is subject to contamination from sewer systems discharge and runoff from agricultural lands. Climate models projects an increase in precipitation and heavy rainfall in this region. This study aimed to determine how daily rainfall causes variation in indicators of pathogen loads, to increase knowledge of variations in river water quality and discuss implications for risk management. Data covering 7 years of daily monitoring of river water turbidity and concentrations of E. coli, Clostridium and coliforms were obtained, and their short-term variations in relation with precipitation were analyzed with time series regression and non-linear distributed lag models. We studied how precipitation effects varied with season and compared different weather stations for predictive ability. Generally, the lowest raw water quality occurs 2 days after rainfall, with poor raw water quality continuing for several more days. A rainfall event of >15 mm/24-h (local 95 percentile) was associated with a three-fold higher concentration of E. coli and 30% higher turbidity levels (lag 2). Rainfall was associated with exponential increases in concentrations of indicator bacteria while the effect on turbidity attenuated with very heavy rainfall. Clear associations were also observed between consecutive days of wet weather and decreased water quality. The precipitation effect on increased levels of indicator bacteria was significant in all seasons. Rainfall elevates microbial risks year-round in this river and freshwater source and acts as the main driver of varying water quality. Heavy rainfall appears to be a better predictor of fecal pollution than water turbidity. An increase of wet weather and extreme events with climate change will lower river water quality even more, indicating greater challenges for drinking water producers, and suggesting better control of sources of pollution.


Mechanism of glycogenolytic action of histamine in rat hepatocytes.
The mechanism by which histamine induces glycogenolysis was investigated in rat hepatocytes. Histamine induced stimulation of glucose output in hepatocytes in a dose-dependent manner. The maximal effect of the glycogenolytic action of histamine, which was approximately 60% of the maximal glucagon action, was obtained at 10(-6) M. These effects were inhibited by H1 receptor antagonists triprolidine hydrochloride and tripelennamine but not by a H2 receptor antagonist cimetidine. Histamine also increased the activity of phosphorylase a. When 10(-6) M histamine and 5 x 10(-9) M glucagon were added simultaneously, the actions of these two agents were additive. In contrast, there was no additivity when 10(-6) M histamine and 10(-8) M angiotensin II were added. Histamine did not increase adenosine 3',5'-cyclic monophosphate at any doses tested but induced a rapid increase in the cytoplasmic free calcium concentration ([Ca2+]c). Histamine increased [Ca2+]c even in the presence of 1 microM extracellular calcium, an observation suggesting that histamine caused calcium release from an intracellular calcium pool(s). When [3H]inositol-labeled hepatocytes were incubated with histamine, radioactivity in the D-myo-inositol trisphosphate fraction was rapidly increased. These results indicate that histamine acts on rat hepatocytes mainly via H1 receptors and stimulates glycogenolysis by activating the calcium messenger system.


The epidemiology of head injuries in English professional rugby union.
To undertake a detailed epidemiological study of head injuries sustained by professional rugby union players in order to define their incidence, nature, severity, and causes. A 3-season prospective cohort design. 13 English Premiership rugby union clubs. 757 male rugby union players. Anatomical location, diagnosis, severity (number of days unavailable for training and match play), injury incident, and incidence of match and training injuries (injuries/1000 player-hours). Playing position, use of headgear, and activity. The overall incidence of match head injury was 6.6 injuries/1000 player-hours, and each injury resulted, on average, in 14 days lost-time. The overall incidence of match concussions was 4.1 injuries/1000 player-hours resulting, on average, in 13 days lost-time. Concussion was the third most common match injury for all players. A large proportion of the players (48%) were able to return to play safely within 7 days. Match concussions were most commonly associated with tackling head-on (28%), collisions (20%), and being tackled head-on (19%). The midfield backs were the playing position at the greatest risk of sustaining a concussion. Only 42% of players were removed from the field of play immediately after a concussion. Foul play was reported by the player to be associated with match head injury in 17% of cases. Mouthguard and headgear usage was associated with a reduced incidence of concussive injury. The overall incidence of training head injury was 0.05 injuries/1000 player-hours, and the overall incidence of training concussions was 0.02 injuries/1000 player-hours. The results showed that rugby union players were exposed to a high risk of noncatastrophic head injury and concussion, particularly whilst tackling and being tackled head-on. In all, 48% of players sustaining a concussion were able to return to play in less than 7 days. The clinical challenge when assessing the potentially concussed player during a game is compounded by the current regulations regarding the permanent replacement of injured players. Injury prevention strategies should be focussed on minimizing the risk and force of direct contact to the head in the tackle.


The stereotactic approach for mapping epileptic networks: a prospective study of 200 patients.
Stereoelectroencephalography (SEEG) is a methodology that permits accurate 3D in vivo electroclinical recordings of epileptiform activity. Among other general indications for invasive intracranial electroencephalography (EEG) monitoring, its advantages include access to deep cortical structures, its ability to localize the epileptogenic zone when subdural grids have failed to do so, and its utility in the context of possible multifocal seizure onsets with the need for bihemispheric explorations. In this context, the authors present a brief historical overview of the technique and report on their experience with 2 SEEG techniques (conventional Leksell frame-based stereotaxy and frameless stereotaxy under robotic guidance) for the purpose of invasively monitoring difficult-to-localize refractory focal epilepsy. Over a period of 4 years, the authors prospectively identified 200 patients with refractory epilepsy who collectively underwent 2663 tailored SEEG electrode implantations for invasive intracranial EEG monitoring and extraoperative mapping. The first 122 patients underwent conventional Leksell frame-based SEEG electrode placement; the remaining 78 patients underwent frameless stereotaxy under robotic guidance, following acquisition of a stereotactic ROSA robotic device at the authors' institution. Electrodes were placed according to a preimplantation hypothesis of the presumed epileptogenic zone, based on a standardized preoperative workup including video-EEG monitoring, MRI, PET, ictal SPECT, and neuropsychological assessment. Demographic features, seizure semiology, number and location of implanted SEEG electrodes, and location of the epileptogenic zone were recorded and analyzed for all patients. For patients undergoing subsequent craniotomy for resection, the type of resection and procedure-related complications were prospectively recorded. These results were analyzed and correlated with pathological diagnosis and postoperative seizure outcomes. The epileptogenic zone was confirmed by SEEG in 154 patients (77%), of which 134 (87%) underwent subsequent craniotomy for epileptogenic zone resection. Within this cohort, 90 patients had a minimum follow-up of at least 12 months; therein, 61 patients (67.8%) remained seizure free, with an average follow-up period of 2.4 years. The most common pathological diagnosis was focal cortical dysplasia Type I (55 patients, 61.1%). Per electrode, the surgical complications included wound infection (0.08%), hemorrhagic complications (0.08%), and a transient neurological deficit (0.04%) in a total of 5 patients (2.5%). One patient (0.5%) ultimately died due to intracerebral hematoma directly ensuing from SEEG electrode placement. Based on these results, SEEG methodology is safe, reliable, and effective. It is associated with minimal morbidity and mortality, and serves as a practical, minimally invasive approach to extraoperative localization of the epileptogenic zone in patients with refractory epilepsy.


On the decomposition of stress and strain tensors into spherical and deviatoric parts.
It is well known that Hooke's law for a linearly elastic, isotropic solid may be written in the form of two relations that involve only the spherical or only the deviatoric parts of the tensors of stress and strain. The example of the linearly elastic, transversely isotropic solid is used to show that this decomposition is not, in general, feasible for linearly elastic, anisotropic solids. The discussion is extended to a large class of work-hardening rigid, plastic solids, and it is shown that the considered decomposition can only be achieved for the incompressible solids of this class.


Coagulation management in patients undergoing neurosurgical procedures.
Management of coagulation in neurosurgical procedures is challenging. In this contest, it is imperative to avoid further intracranial bleeding. Perioperative bleeding can be associated with a number of factors, including anticoagulant drugs and coagulation status but is also linked to the characteristic and the site of the intracranial disorder. The aim of this review will be to focus primarily on the new evidence regarding the management of coagulation in patients undergoing craniotomy for neurosurgical procedures. Antihemostatic and anticoagulant drugs have shown to be associated with perioperative bleeding. On the other hand, an increased risk of venous thromboembolism and hypercoagulative state after elective and emergency neurosurgery, in particular after brain tumor surgery, has been described in several patients. To balance the risk between thrombosis and bleeding, it is important to be familiar with the perioperative changes in coagulation and with the recent management guidelines for anticoagulated patients undergoing neurosurgical procedures, in particular for those taking new direct anticoagulants. We have considered the current clinical trials and literature regarding both safety and efficacy of deep venous thrombosis prophylaxis in the neurosurgical population. These were mainly trials concerning both elective surgical and intensive care patients with a poor grade intracranial bleed or multiple traumas with an associated severe traumatic brain injury (TBI). Coagulation management remains a major issue in patients undergoing neurosurgical procedures. However, in this field of research, literature quality is poor and further studies are necessary to identify the best strategies to minimize risks in this group of patients.


Work group diversity and group performance: an integrative model and research agenda.
Research on the relationship between work group diversity and performance has yielded inconsistent results. To address this problem, the authors propose the categorization-elaboration model (CEM), which reconceptualizes and integrates information/decision making and social categorization perspectives on work-group diversity and performance. The CEM incorporates mediator and moderator variables that typically have been ignored in diversity research and incorporates the view that information/decision making and social categorization processes interact such that intergroup biases flowing from social categorization disrupt the elaboration (in-depth processing) of task-relevant information and perspectives. In addition, the authors propose that attempts to link the positive and negative effects of diversity to specific types of diversity should be abandoned in favor of the assumption that all dimensions of diversity may have positive as well as negative effects. The ways in which these propositions may set the agenda for future research in diversity are discussed.


A programme for the construction of a lambda phage.
Infection of a sensitive host by the lambdoid coliphages can cause death of the bacterial cell by lysis or can lead to a lysogenic cell, with the viral DNA stably integrated into the host chromosome. These alternative responses both require the coordination of several host and phage functions, and lambda infection follows a well controlled developmental plan. The lytic and lysogenic pathways of lambdoid infection are reviewed, with emphasis on the variety of control mechanisms involved in the commitment to a particular pathway.


Assessment of mineralocorticoid activity in the rabbit colon.
Analyses of [3H]corticosteroid binding sites in distal colon indicated high-affinity binding sites or receptors for both [3H]aldosterone (Type I, Kd = 6.5 X 10(-9) M) and [3H]dexamethasone (Type II, Kd = 5.5 X 10(-8) M). The relative affinity of dexamethasone (D) was 1/20 that of aldosterone (A) for Type I sites and the affinity of A for Type II sites was 1/50 that of D at 37 degrees C. Citrate synthase (CS) activity was assayed and found to be reduced in enterocytes harvested from adrenalectomized (ADX) vs. normal colon segments (0.24 vs. 0.44 U/mg protein, P less than 0.025). Aldosterone (10 micrograms/kg body wt) increased CS at 2 h to a level intermediate between normal and ADX animals and thus not significantly different from either group, but was significantly increased over ADX + D values. Transmural potential difference was increased by 10(-8) M A but not by 10(-8) M D. Since both steroids enhanced short-circuit current at this concentration, the dichotomy of the glucocorticoid vs. mineralocorticoid results can be best explained by the pronounced effect of D on resistance (R) across the tissue (R at 4 h + D was 50% that of paired controls). These findings would suggest that the rabbit distal colon is a target segment for both mineralocorticoids and glucocorticoids. Furthermore, as in the kidney, the two steroids may play coordinated but, perhaps in some way, unique roles in the regulation of transport.


Protein complex identification by supervised graph local clustering.
Protein complexes integrate multiple gene products to coordinate many biological functions. Given a graph representing pairwise protein interaction data one can search for subgraphs representing protein complexes. Previous methods for performing such search relied on the assumption that complexes form a clique in that graph. While this assumption is true for some complexes, it does not hold for many others. New algorithms are required in order to recover complexes with other types of topological structure. We present an algorithm for inferring protein complexes from weighted interaction graphs. By using graph topological patterns and biological properties as features, we model each complex subgraph by a probabilistic Bayesian network (BN). We use a training set of known complexes to learn the parameters of this BN model. The log-likelihood ratio derived from the BN is then used to score subgraphs in the protein interaction graph and identify new complexes. We applied our method to protein interaction data in yeast. As we show our algorithm achieved a considerable improvement over clique based algorithms in terms of its ability to recover known complexes. We discuss some of the new complexes predicted by our algorithm and determine that they likely represent true complexes. Matlab implementation is available on the supporting website: www.cs.cmu.edu/~qyj/SuperComplex.


The role of category coherence in experience-based prediction.
Both real-world category knowledge and instance-based sample data are often available as sources of inductive inference. In three experiments using natural social categories, we test the influence of generalcategory knowledge on the use of category instances to make property inductions both to other category members and to others in the population. We find that a category's coherence--the extent to which its features are interrelated through prior knowledge (Murphy & Medin, 1985)--influences inductions positively to new category members and negatively to the population. This effect of coherence is strongest with small as compared with large samples of instances. The results are interpreted from both similarity and explanation-based perspectives.


Nutrition and preparation of blenderized tube feeding in children and adolescents with neurological impairment: A scoping review.
The use of homemade tube feeding formula has become increasingly popular for children requiring enteral nutrition. This project aimed to investigate nutrition and preparation of blenderized tube feeding in the field of children and adolescents with neurological impairment. A scoping review was performed using established methodologies. In January 2021, we searched PubMed, Embase, CINAHL Complete, the Cochrane Central Register of Controlled Trials, and gray literature to identify relevant articles. Twenty-two papers were included describing the composition of food items, preparation procedures, and food safety. No randomized controlled trials and only a few prospective studies were included. A broad variety of food items from all food groups and many examples of recipes were presented. Most recipes provided 1.0 kcal/ml but tended to contain less energy and nutrients than expected, which could be due to preparation issues, such as sieving and the high viscosity of the blend. Preparation requires a commercial-grade household blender and diligence to ensure thorough household hygiene for adequate food safety. This review revealed practical experience in the nutrition and preparation aspects of blenderized tube feeding but minimal empirical evidence. Multiple examples of the composition of food items and preparation procedures for blenderized tube feeding were found, but uncertainty regarding the ideal composition or preparation was also exposed. The future of blenderized tube feeding would benefit from clinically tested recipes that include an evaluation of nutrients, viscosity, and microbial contamination, as well as the effect of the food's appearance and scent on the target group.


Adverse events with radiographic contrast agents: results of the SCVIR Contrast Agent Registry.
To determine prospectively the incidence of adverse events in angiography related to contrast agents, and the relative incidence of events with use of high-osmolality contrast agents (HOCAs) and low-osmolality contrast agents (LOCAs) and to ascertain if risk factors can help identify increased risk of an adverse event and patients likely to benefit from use of LOCAs. From 26 high-volume institutions, data were collected on every patient who underwent angiography from July 1, 1990, to June 30, 1992. Information included demographic and risk factors, general medical status, previous administration of contrast media, procedural information, occurrence and characteristics of all adverse events up to 12 hours after procedures, and relation to contrast agents, treatment, and outcome. In 60,891 patients, there were 75,616 studies, 56% with nonionic LOCAs, 8% with the ionic LOCA, and 36% with HOCAs. Most major risk factors correlated with an increased incidence of adverse events related to contrast media. Incidence of these adverse events varied with type of procedure, with a higher incidence associated with cardiac and interventional procedures. Overall adverse events related to contrast media and those for which treatment was necessary were significantly increased (P < .001) with use of HOCAs for all but arterial interventional procedures. Serious adverse events were not different between the two classes of agents except for cardiac procedures. Previous reaction to contrast medium was the most important risk factor in prediction of an adverse event. The safety benefit of use of LOCAs is limited. Patients most likely to benefit are those with a previous reaction or more than one other major risk factor. Selective use of LOCAs is an appropriate strategy.


Analyzing the movement of the Nauplius 'Artemia salina' by optical tracking of plasmonic nanoparticles.
We demonstrate how optical tweezers may provide a sensitive tool to analyze the fluidic vibrations generated by the movement of small aquatic organisms. A single gold nanoparticle held by an optical tweezer is used as a sensor to quantify the rhythmic motion of a Nauplius larva (Artemia salina) in a water sample. This is achieved by monitoring the time dependent displacement of the trapped nanoparticle as a consequence of the Nauplius activity. A Fourier analysis of the nanoparticle's position then yields a frequency spectrum that is characteristic to the motion of the observed species. This experiment demonstrates the capability of this method to measure and characterize the activity of small aquatic larvae without the requirement to observe them directly and to gain information about the position of the larvae with respect to the trapped particle. Overall, this approach could give an insight on the vitality of certain species found in an aquatic ecosystem and could expand the range of conventional methods for analyzing water samples.


In vivo phosphorylation of FtsZ2 in Arabidopsis thaliana.
The tubulin-like FtsZ protein initiates assembly of the bacterial and plastid division machineries. In bacteria, phosphorylation of FtsZ impairs GTPase activity, polymerization and interactions with other division proteins. Using a proteomics approach, we have shown that AtFtsZ2 is phosphorylated in vivo in Arabidopsis and that PGK1 (phosphoglycerate kinase 1) interacts with AtFtsZ2 in planta, suggesting a possible role in FtsZ phosphorylation.


Usefulness of Platelet-to-Lymphocyte Ratio as a Marker of Sarcopenia for Critical Limb Threatening Ischemia.
Sarcopenia is a factor of poor prognosis for patients with critical limb threatening ischemia (CLTI), but its diagnosis requires imaging measurements and is time consuming. We investigated whether preoperative platelet-to-lymphocyte ratio (PLR) could be an easy and rapid marker of sarcopenia. Patients treated for CLTI between January 2019 and July 2019 were included in this single-center retrospective study. Sarcopenia was defined by a psoas muscle index (PMI) <5.5 cm2/m2 in men, and <4.0 cm2/m2 in women. PLR was calculated for all patients based on their systematic preoperative blood analysis. The diagnostic power of PLR was analyzed through a receiver operating characteristic (ROC) curve. Early outcomes of sarcopenic patients in terms of 30-day mortality and 30-day morbidity were retrieved. Sixty-four patients were included in the study: 48 nonsarcopenic patients (mean PMI 7.34 cm2/m2; interquartile range [IQR] 6.58-8.01) and 16 sarcopenic patients (mean PMI 4.30 cm2/m2; IQR 3.45-5.17). No difference was found between both groups regarding patient demographics, clinical characteristics, cardiovascular risk factors, comorbidities, or revascularization modalities. PLR was significantly higher in the sarcopenic group (mean 332.1; IQR 158.2-320.7) compared with the nonsarcopenic group (mean 204.6; IQR 133.8-265.6) (P = 0.02). A PLR value ≥292.5 was shown to be a diagnostic marker for sarcopenia based on the ROC curve (sensitivity 31.3%, specificity 91.7%). Thirty-day mortality was 12.5% in the sarcopenic group and 2.1% in the nonsarcopenic group (P = 0.15); 30-day morbidity was 56.3% in the sarcopenic group and 10.4% in the nonsarcopenic group (P < 0.001). PLR might help identifying a subgroup of CTLI patients associated with poor prognosis but does not seem appropriate to be used as a marker of sarcopenia given its low sensitivity.


Combined physical, chemical and biological factors shape Alexandrium ostenfeldii blooms in The Netherlands.
Harmful algal blooms (HABs) are globally expanding, compromising water quality worldwide. HAB dynamics are determined by a complex interplay of abiotic and biotic factors, and their emergence has often been linked to eutrophication, and more recently to climate change. The dinoflagellate Alexandrium is one of the most widespread HAB genera and its success is based on key functional traits like allelopathy, mixotrophy, cyst formation and nutrient retrieval migrations. Since 2012, dense Alexandrium ostenfeldii blooms (up to 4500cellsmL-1) have recurred annually in a creek located in the southwest of the Netherlands, an area characterized by intense agriculture and aquaculture. We investigated how physical, chemical and biological factors influenced A. ostenfeldii bloom dynamics over three consecutive years (2013-2015). Overall, we found a decrease in the magnitude of the bloom over the years that could largely be linked to changing weather conditions during summer. More specifically, low salinities due to excessive rainfall and increased wind speed corresponded to a delayed A. ostenfeldii bloom with reduced population densities in 2015. Within each year, highest population densities generally corresponded to high temperatures, low DIN:DIP ratios and low grazer densities. Together, our results demonstrate an important role of nutrient availability, absence of grazing, and particularly of the physical environment on the magnitude and duration of A. ostenfeldii blooms. Our results suggest that predicted changes in the physical environment may enhance bloom development in future coastal waters and embayments.


Insights Into the Mechanisms of Brain Endothelial Erythrophagocytosis.
The endothelial cells which form the inner cellular lining of the vasculature can act as non-professional phagocytes to ingest and remove emboli and aged/injured red blood cells (RBCs) from circulation. We previously demonstrated an erythrophagocytic phenotype of the brain endothelium for oxidatively stressed RBCs with subsequent migration of iron-rich RBCs and RBC degradation products across the brain endothelium in vivo and in vitro, in the absence of brain endothelium disruption. However, the mechanisms contributing to brain endothelial erythrophagocytosis are not well defined, and herein we elucidate the cellular mechanisms underlying brain endothelial erythrophagocytosis. Murine brain microvascular endothelial cells (bEnd.3 cells) were incubated with tert-butyl hydroperoxide (tBHP, oxidative stressor to induce RBC aging in vitro)- or PBS (control)-treated mouse RBCs. tBHP increased the reactive oxygen species (ROS) formation and phosphatidylserine exposure in RBCs, which were associated with robust brain endothelial erythrophagocytosis. TNFα treatment potentiated the brain endothelial erythrophagocytosis of tBHP-RBCs in vitro. Brain endothelial erythrophagocytosis was significantly reduced by RBC phosphatidylserine cloaking with annexin-V and with RBC-ROS and phosphatidylserine reduction with vitamin C. Brain endothelial erythrophagocytosis did not alter the bEnd.3 viability, and tBHP-RBCs were localized with early and late endosomes. Brain endothelial erythrophagocytosis increased the bEnd.3 total iron pool, abluminal iron levels without causing brain endothelial monolayer disruption, and ferroportin levels. In vivo, intravenous tBHP-RBC injection in aged (17-18 months old) male C57BL/6 mice significantly increased the Prussian blue-positive iron-rich lesion load compared with PBS-RBC-injected mice. In conclusion, RBC phosphatidylserine exposure and ROS are key mediators of brain endothelial erythrophagocytosis, a process which is associated with increased abluminal iron in vitro. tBHP-RBCs result in Prussian blue-positive iron-rich lesions in vivo. Brain endothelial erythrophagocytosis may provide a new route for RBC/RBC degradation product entry into the brain to produce iron-rich cerebral microhemorrhage-like lesions.


OPTN/SRTR 2013 Annual Data Report: pancreas.
Pancreas listings and transplants decreased during the past decade, most notably pancreas after kidney transplants. Center-reported outcomes of pancreas transplant across all groups, short-term and long-term, improved during the same period. Changes to the pancreas allocation system creating an efficient, uniform national system will be implemented in late 2014. Pancreas-alone and simultaneous pancreas-kidney (SPK) candidates will form a single match-run list with priority to most SPK candidates ahead of kidney-alone candidates to decrease waiting times for SPK candidates, given their higher waitlist mortality compared with nondiabetic kidney transplant candidates. The changes are expected to eliminate local variability, providing more consistent pancreas allocation nationwide. Outcomes after pancreas transplant are challenging to interpret due to lack of a uniform definition of graft failure. Consequently, SRTR has not published data on pancreas graft failure for the past 2 years. The Organ Procurement and Transplantation Network Pancreas Transplantation Committee is working on a definition that could provide greater validity for future outcomes analyses. Challenges in pancreas transplantation include high risk of technical failures, rejection (early and late), and surgical complications. Continued outcome improvement and innovation has never been more critical, as alternatives such as islet transplant and artificial pancreas move closer to clinical application.


Acupuncture analgesia. I. Review of clinical studies.
Recent clinical studies, both uncontrolled and controlled, of the use of acupuncture for the relief of chronic pain are reviewed. The success rate demonstrated by controlled trials is markedly lower than that claimed in reports of the clinical use of acupuncture in which no attempt is made to control factors which influence the placebo response. Although on balance of evidence it appears that acupuncture has a place in the management of chronic pain resistant to orthodox therapeutic measures, more research is needed both to establish objective criteria for the assessment of the effect, or lack of effect, of acupuncture on chronic pain as well as to delineate clearly the patient groups and the conditions where acupuncture analgesia is indicated.


Observer variation in the radiographic classification of ankle fractures.
We recorded inter- and intra-observer variations in the classification of ankle fractures by the Lauge Hansen and Weber systems. Radiographs of 94 patients were classified independently by four observers. The observer variation was calculated by kappa statistics, which corrects the obtained values for the agreement expected by chance. There was an acceptable level of agreement for the overall classification into both systems. For the staging of supination-adduction and supination-eversion fractures in the Lauge Hansen system the agreement was poor. The results indicate that future classification systems should be subject to reliability analysis before they are accepted.


Saturated hydrocarbons in bovine liver.
A homologous series of n-alkanes (C(14)-C(33)) and two isoprenoid hydrocarbons, 2,6,10,14-tetramethylhexadecane (phytane) and 2,6,10,14-tetramethylpentadecane (pristane) have been identified in bovine liver. Another branched but non-isoprenoid alkane and three isomers of molecular formula C(20)H(40) were partially identified. Phytane and the C(18)-C(22) and C(29)-C(33)n-alkanes were found to be the major components in liver, suggesting that at least the main hydrocarbon components were derived from various plants in the diet. The hydrocarbons were separated and identified by a series of steps involving solvent extraction, saponification, elution chromatography on alumina and silica gel columns, molecular sieving and by infrared and ultraviolet spectroscopy, followed by combined capillary gas chromatography-mass spectrometry.


Impaired fertility and FSH synthesis in gonadotrope-specific Foxl2 knockout mice.
Impairments in pituitary FSH synthesis or action cause infertility. However, causes of FSH dysregulation are poorly described, in part because of our incomplete understanding of mechanisms controlling FSH synthesis. Previously, we discovered a critical role for forkhead protein L2 (FOXL2) in activin-stimulated FSH β-subunit (Fshb) transcription in immortalized cells in vitro. Here, we tested the hypothesis that FOXL2 is required for FSH synthesis in vivo. Using a Cre/lox approach, we selectively ablated Foxl2 in murine anterior pituitary gonadotrope cells. Conditional knockout (cKO) mice developed overtly normally but were subfertile in adulthood. Testis size and spermatogenesis were significantly impaired in cKO males. cKO females exhibited reduced ovarian weight and ovulated fewer oocytes in natural estrous cycles compared with controls. In contrast, ovaries of juvenile cKO females showed normal responses to exogenous gonadotropin stimulation. Both male and female cKO mice were FSH deficient, secondary to diminished pituitary Fshb mRNA production. Basal and activin-stimulated Fshb expression was similarly impaired in Foxl2 depleted primary pituitary cultures. Collectively, these data definitively establish FOXL2 as the first identified gonadotrope-restricted transcription factor required for selective FSH synthesis in vivo.


Effect of altitude exposure on induction of streptococcal endocarditis in young and middle-aged rats.
Young (age 2 months) and middle-aged (age 10 month) rats were injected once with a culture of Streptococcus sanguis and exposed for 24 h to 7620 m altitude. At 6 d 54% of the exposed and 30% of the unexposed middle-aged rats had bacterial endocarditis. Myocarditis developed in 63% of the injected exposed rats of both ages, in 11% of the injected unexposed middle-aged rats, and in none of the unexposed young adults. Interstitial nephritis was found in 46-66% of the injected, unexposed young and middle-aged rats and in 70-86% of the injected, exposed young and middle-aged rats, respectively. About 95% of all injected rats survived 6 d. No evidence of hemoconcentration was found. The increase in cardiac disease induced by altitude was probably due to deleterious effects of hypoxia on the myocardium, and cellular defenses, and to physiological and possible immunological changes associated with aging.


Interpretation of single progesterone measurement in diagnosis of anovulation and defective luteal phase: observations on analysis of the normal range.
Single serum progesterone determinations were made in 79 apparently normal women with a regular menstrual cycle. A normal range (40 subjects) was derived from the concentrations in the follicular phase and used to define an "anovular" range for luteal phase values (nine out of 39 subjects). The remaining luteal phase values were used to construct an "ovular" range for the luteal phase and, within this range, to define a group of values (less than the 20th centile) which could be described as a "defective luteal phase." The cut off limits between ovular and anovular and between normal and defective luteal phases were respectively two and four times the follicular phase median. It is proposed that the numerical findings of this study may be used as a rule of thumb for defining normality and abnormality from a single serum progesterone determination.


[Abdominal bipedicled subepidermal vascular-network flap in repair of degloving injury of hand].
To evaluate the clinical result of repairing degloving injury of hand with abdominal bipedicled subdermal vascular-network flap in emergence. From 1994 to 1997, 19 cases with degloving injury of hand were treated with two flaps, one flaps with the inferior epigastric artery as pedicle, another with superficial epigastric artery. The two skin flaps were designed oppositely to cover the injured hands. All the flaps survived. Followed up for 1 to 3 years, the contour of hands were fine. Extension of fingers was normal and opponers of thumbs was good. Range of flexion of metacarpo-phalangeal joints was from 45 to 60 degrees, and the proximal interphalangeal joints was from 10 to 25 degrees. For the subdermal vascular network, the bipedicled flaps of abdomen in repair of degloving injury of hand have sufficient blood supply, strong resistance to infection, high survival rate, and good contour.


Medical management of diabetic foot infections.
Diabetic foot infections (DFIs) are a commonly encountered medical problem. They are associated with an increased frequency and length of hospitalization and risk for lower-extremity amputation. Furthermore, they have substantial economic consequences. Patients with diabetes mellitus are particularly susceptible to foot infections because of neuropathy, vascular insufficiency, and diminished neutrophil function. The approach to managing DFIs starts with determining if an infection exists. If an infection exists, then the type, severity, extent of infection, and risk factors for resistant organisms should be determined through history, physical examination, and additional laboratory and radiological testing. Optimal management requires surgical debridement, pressure offloading, effective antibiotic therapy, wound care and moisture, maintaining good vascular supply, and correction of metabolic abnormalities, such as hyperglycemia, through a multidisciplinary team. Empiric antibiotics for DFIs vary based on the severity of the infection, but must include anti-staphylococcal coverage.


The breakfast tolerance test: a return to physiology.
The effects of a standardized breakfast on blood glucose, plasma free fatty acid and alpha-amino nitrogen levels, plasma insulin, glucagon and cortisol concentrations as well as serum growth hormone circulating levels are described in a group of six healthy, young, male volunteers. It is suggested to the clinician to use this "breakfast tolerance test" or B.T.T. as a simple means to reconsider on physiological grounds some widely accepted pathophysiological concepts derived from highly unphysiological procedures such as the oral glucose tolerance test or O.G.T.T.


The role of fingers in number processing in young children.
The aim of the present study was to investigate the relationship between finger counting and numerical processing in 4-7-year-old children. Children were assessed on a variety of numerical tasks and we examined the correlations between their rates of success and their frequency of finger use in a counting task. We showed that children's performance on finger pattern comparison and identification tasks did not correlate with the frequency of finger use. However, this last variable correlated with the percentages of correct responses in an enumeration task (i.e., Give-N task), even when the age of children was entered as a covariate in the analysis. Despite this correlation, we showed that some children who never used their fingers in the counting task were able to perform optimally in the enumeration task. Overall, our results support the conclusion that finger counting is useful but not necessary to develop accurate symbolic numerical skills. Moreover, our results suggest that the use of fingers in a counting task is related to the ability of children in a dynamic enumeration task but not to static tasks involving recognition or comparison of finger patterns. Therefore, it could be that the link between fingers and numbers remain circumscribed to counting tasks and do not extent to static finger montring situations.


Coenzyme Q10 supplementation provides mild symptomatic benefit in patients with Parkinson's disease.
Features of Parkinson's disease (PD) include oxidative stress, nigral mitochondrial complex I deficiency and visual dysfunction, all of which are also associated with coenzyme Q(10) (CoQ(10)) deficiency. The objective of this monocenter, parallel group, placebo controlled, double-blind trial was to determine the symptomatic response of daily oral application of 360 mg CoQ(10) lasting 4 weeks on scored PD symptoms and visual function, measured with the Farnsworth-Munsell 100 Hue test (FMT), in 28 treated and stable PD patients. CoQ(10) supplementation provided a significant (P=0.01) mild symptomatic benefit on PD symptoms and a significantly (F((1,24))=8.48, P=0.008) better improvement of FMT performance compared with placebo. Our results indicate a moderate beneficial effect of oral CoQ(10) supplementation in PD patients.


The clotting defect in SLE.
Haemorrhagic complications, though uncommon in SLE, may be life-threatening in individual patients, and they require treatment along appropriate lines. Thrombotic problems are more commonly encountered, and are a significant cause of morbidity and mortality. Not only is clinical thrombosis important in SLE, but there is increasing evidence that low-grade coagulopathy contributes substantially to many of the pathological features seen in lupus. The mechanisms involved in the pathogenesis of thrombosis have been discussed and their possible interrelationship is summarized in Figure 4, though it remains speculation that low-grade coagulopathy predisposes to clinical thrombosis. Several of these mechanisms may be operating during periods of disease activity; this was suggested in a recent study of clinical and histological features in SLE (Kant et al, 1981). The study was designed to look at the prevalence of glomerular thrombosis in SLE, and its significance as a histological feature. A striking association was observed between the presence of a circulating anticoagulant and the appearance of glomerular thrombosis on renal biopsy. Also, factor VIII levels were significantly increased and circulating platelets decreased in association with "active' histological features. On later re-biopsy glomerulosclerosis was a much more common finding if the first biopsy showed thrombosis, suggesting that thrombosis is a marker of more severe disease activity and inflammation. A greater understanding of the mechanisms promoting thrombosis will undoubtedly provide insight into the pathogenesis of SLE, as well as suggesting new therapeutic possibilities.


The Application of "Bone Window" Technique in Endodontic Microsurgery.
Endodontic microsurgery for a tooth with a large periapical lesion and an intact cortical plate may necessitate the removal of extra bone and cause delayed or unfavorable healing. In such cases, the "bone window" technique offers excellent exposure to the operative field and preserves cortical bone without placing any additional graft material. In the reported cases, root-end surgery was performed on the maxillary and mandibular molars with a large periapical lesion. The bone window was fashioned with the aid of thin osteotomy instruments and repositioned to the original site at the end of the procedure, which resulted in minimizing bone loss and acted as an autologous graft for the surgical site. At 12- and 16-month follow-ups, clinical examinations and cone-beam computed tomographic scans revealed the healing of the lesion without symptoms or complications. Cone-beam computed tomographic imaging was used as a presurgical assessment tool for indication selection and precise design of the bone window. The use of a bone window in endodontic microsurgery, which appears to be a reliable technique, should be the method of preference when the large lesion is deeply positioned between intact buccal and lingual cortices.


ACR Appropriateness Criteria® Radiologic Management of Portal Hypertension.
Cirrhosis is a heterogeneous disease that cannot be studied as a single entity and is classified in two main prognostic stages: compensated and decompensated cirrhosis. Portal hypertension, characterized by a pathological increase of the portal pressure and by the formation of portal-systemic collaterals that bypass the liver, is the initial and main consequence of cirrhosis and is responsible for the majority of its complications. A myriad of treatment options exists for appropriately managing the most common complications of portal hypertension, including acute variceal bleeding and refractory ascites. The American College of Radiology Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed annually by a multidisciplinary expert panel. The guideline development and revision include an extensive analysis of current medical literature from peer reviewed journals and the application of well-established methodologies (RAND/UCLA Appropriateness Method and Grading of Recommendations Assessment, Development, and Evaluation or GRADE) to rate the appropriateness of imaging and treatment procedures for specific clinical scenarios. In those instances where evidence is lacking or equivocal, expert opinion may supplement the available evidence to recommend imaging or treatment.


[Role of cerebral oximetry in prediction of hyperperfusion syndrome after carotid endarterectomy].
The purpose of the study was to carry out a retrospective analysis of alterations in regional oxygenation (rSO2) of the brain at all stages of carotid endarterectomy (CEA) in order to detect predictors of ischaemia and hyperperfusion of the brain. In a total of 169 patients during CEA under general anaesthesia we registered rSO2 after induction of narcosis, prior to carotid artery cross-clamping, at 2 minutes and each 10 minutes after cross-clamping, before re-establishing blood flow, at 2 and 5 minutes thereafter, and finally at the end of the operation. We evaluated the baseline parameters and intraoperative alterations in regional oxygenation depending on clinical and instrumental data. We also carried out a multivariate regression analysis in order to reveal significant risk factors for ischaemia and hyperperfusion. We analysed the ROC curves in order to determine the threshold value of a decrease and increase of rSO2 in ischaemia and hyperperfusion of the brain. After the multivariate analysis, an independent predictor turned out to be grade III arterial hypertension (OR 9.5, 95% CI: 1.1-82.7). It was revealed that the most significant predictor of the development of hyperperfusion syndrome was the absolute increase in rSO2 after re-establishing blood flow by more than 11.3. Sensitivity, specificity, positive and negative prognostic value of the parameter revealed amounted to 87.4, 83, 35 and 98.4%, respectively. The multivariate analysis performed demonstrated that only long-standing grade III arterial hypertension was a significant risk factor for the development of hyperperfusion syndrome after CEA. In our study, the most accurate parameter for the Fore-Sight oximeter was the maximum increase of the absolute value of rSO2 after re-establishing blood flow above 11.3 Cerebral oximetry is a non-invasive, simple to use and interpreter method making it possible to evaluate not only ischaemia but hyperperfusion of the brain during CEA and in the early postoperative period.


Gut Microbiota Differences According to Ultra-Processed Food Consumption in a Spanish Population.
Ultra-processed foods (UPFs) consumption could affect gut microbiota diversity and profile. We aimed to evaluate the effects of UPFs on microbiota, considering the role of sex. The consumption of UPFs (using NOVA criteria) was assessed with a validated 137-item food-frequency questionnaire. Participants (n = 359) were classified into less than three servings per day (n = 96) of UPFs and more than five (n = 90). Women and men were subclassified following the same criteria. 16S rRNA sequencing was performed from DNA fecal samples, and differences in microbiota were analyzed using EdgeR. The relationship between UPFs and bacteria was assessed by Spearman correlation and comparison of tertiles of consumption. Women who consumed more than five servings/day of UPFs presented an increase in Acidaminococcus, Butyrivibrio, Gemmiger, Shigella, Anaerofilum, Parabacteroides, Bifidobacterium, Enterobacteriales, Bifidobacteriales and Actinobacteria and a decrease in Melainabacter and Lachnospira. Bifidobacterium, Bifidobacteriales and Actinobacteria was positively associated with pizza and Actinobacteria with industrially processed dairy in women. Men who consumed more than five servings/day presented an increase of Granulicatella, Blautia, Carnobacteriaceae, Bacteroidaceae, Peptostreptococcaceae, Bacteroidia and Bacteroidetes and a decrease of Anaerostipes and Clostridiaceae. Bacteroidia and Bacteroidetes correlated positively with industrially processed meat. This study suggests that UPFs may affect microbiota composition differently in women and men.


Redefining primary hyperaldosteronism as "The Syndrome of Inappropriate Aldosterone Secretion (SIALDS)": A common but unrecognized cause of hypertension.
The current screening and diagnostic recommendations for detecting Primary Hyperaldosteronism (PHA) focus on diagnosing the more severe and overt instances of renin-independent aldosterone production. However, milder forms of autonomous aldosterone secretion have been demonstrated to exist below the diagnostic thresholds of current PHA guidelines, and associate with clinically relevant cardiovascular risk. PHAencompasses a spectrum of renin independent aldosterone production, progressing from a subclinical state in normotensives to a full-blown clinical syndrome representing the resistant hypertension population. The authors propose the Syndrome of Inappropriately Elevated Aldosterone Secretion (SIALDS) concept as a potential new paradigm for understanding and diagnosing PHA and expanded diagnostic approach to improve early detection even in well-controlled hypertension. The authors also delve into the impact of treatments, including mineralocorticoid receptor antagonists and emerging aldosterone synthase inhibitors. Furthermore, The authors outline future research directions, proposing clinical trials to investigate the long-term identification and treatment outcomes of SIALDS.


Warehouse Control of Aspergillus flavus Link and A. parasiticus Speare on Peanuts ( Arachis hypogaea ) by Phosphine Fumigation and its Effect on Aflatoxin Production.
Six stacks of 36 bags containing ca. 50 kg of unshelled peanuts with moisture contents in the range of 18.0 to 21.0% (wet basis) (average 19.3%) were formed in a commercial warehouse located in a peanut-producing area in São Paulo, Brazil. Three stacks were fumigated with phosphine for 7 days. An initial dose of 3.0 g of aluminum phosphide per m3 was applied. A second and a third application of the same dose were carried out 24 and 144 h later. Before fumigation, infection by Aspergillus flavus and/or A. parasiticus was either not detected in the peanut mass or was very low (1 to 13%), but contamination with aflatoxins (up to 191 μg/kg) was found in all stacks. After the fumigation treatments, a striking increase in infection was observed in the nonfumigated stacks (73 to 100% infected kernels) while in the fumigated stacks, A. flavus and/or A. parasiticus were either not detected or were isolated in insignificant amounts, indicating that phosphine was able to control fungal development in spite of the high moisture content of the kernels. After the fumigation period, the contamination levels of aflatoxins in the treated stacks remained unchanged, while the untreated stacks showed a staggering increase (up to 10,000 μg/kg of peanuts). After a month, however, no difference was observed in aflatoxin contamination and infection by A. flavus and A. parasiticus between the untreated and the treated stacks.


Delayed eosinophil apoptosis in asthma.
Eosinophilic inflammation of the airways is a key characteristic of asthma. A defect in apoptosis might contribute to the chronic tissue eosinophilia associated with asthma. Our purpose was to examine whether the rate of apoptosis differs between peripheral blood eosinophils from asthmatic patients and healthy volunteers. Peripheral blood was obtained from volunteers with asthma and from control volunteers. Eosinophils were isolated by CD16-negative selection to >99% purity and were cultured for 48 hours. The number of apoptotic eosinophils in the culture was assessed by flow cytometric analysis of relative DNA content in propidium iodide-stained cells. Eosinophil apoptosis is expressed as apoptosis index (number of apoptotic cells/total number of cells). Eosinophils from asthmatic patients not taking steroid medication survived longer (apoptosis index 0.25) than those of healthy control subjects (apoptosis index 0.40, P <.05). In contrast, the rate of apoptosis in eosinophils from patients concurrently taking steroids (apoptosis index 0.46) is higher than that of those not using steroids (P <.01) and not different from that of healthy subjects. To assess whether endogenous IL-3, IL-5, and GM-CSF production contributes to the delayed eosinophil apoptosis, the effects of the corresponding neutralizing antibodies were studied on eosinophil longevity. Neutralization of GM-CSF, but not of IL-3 or IL-5, increased slightly but significantly (P <.01) the rate of apoptosis in eosinophils obtained from patients with asthma. To assess whether beta(2)-agonist medication could contribute to the observed differences, we determined the in vitro effects of albuterol, fenoterol, and salmeterol on eosinophil apoptosis. All beta(2)-agonists inhibited eosinophil apoptosis by 12% to 19%. A possibility existed that a prior in vivo exposure to IL-5, GM-CSF, or beta(2)-agonists would explain the observed difference. To study this, eosinophils were incubated with GM-CSF, IL-5, and albuterol for 2 to 3 hours, followed by washout of the added compounds, and were subsequently cultured for 48 hours. However, an exposure to GM-CSF (7 pmol/L) or IL-5 (10 pmol/L) for 15 to 180 minutes was not a sufficient signal to prevent eosinophil apoptosis. In contrast, exposure to albuterol (100 nmol/L) for 120 minutes was sufficient to induce a significant (P <.05) decrease in eosinophil apoptosis. The results suggest that eosinophil apoptosis is delayed in asthma. This delay may be partly explained by production of GM-CSF. The in vitro effects of beta(2)-agonists suggest that beta(2)-agonist use might contribute to the prolonged eosinophil survival through inhibition of apoptosis and thus may worsen eosinophilia in asthmatic patients. Use of inhaled glucocorticoids seems to totally reverse the delayed eosinophil apoptosis in asthma.


Significance analysis of time course microarray experiments.
Characterizing the genome-wide dynamic regulation of gene expression is important and will be of much interest in the future. However, there is currently no established method for identifying differentially expressed genes in a time course study. Here we propose a significance method for analyzing time course microarray studies that can be applied to the typical types of comparisons and sampling schemes. This method is applied to two studies on humans. In one study, genes are identified that show differential expression over time in response to in vivo endotoxin administration. By using our method, 7,409 genes are called significant at a 1% false-discovery rate level, whereas several existing approaches fail to identify any genes. In another study, 417 genes are identified at a 10% false-discovery rate level that show expression changing with age in the kidney cortex. Here it is also shown that as many as 47% of the genes change with age in a manner more complex than simple exponential growth or decay. The methodology proposed here has been implemented in the freely distributed and open-source edge software package.