Search

OBJECTIVES: The aims of the present study were to (i) cross-culturally adapt a Danish consensus version of the 10-item Perceived Stress Scale (PSS-10) and (ii) evaluate its psychometric properties in terms of agreement, reliability, validity, responsiveness, and interpretability among patients with work-related stress complaints. METHODS: A consensus-building process was performed involving the authors of the three previous Danish translations and the consensus version was back-translated into English and pilot-tested. Psychometric properties of the final version were examined in a sample of 64 patients with work-related stress complaints. RESULTS: The face validity, reliability, and internal consistency of the Danish consensus version of the PSS-10 were satisfactory, and convergent construct validity was confirmed. Receiver operating characteristic (ROC) curves of the change scores showed that the ability of the PSS-10 to correctly classify patients as improved or unchanged according to the patients’ own judgment was acceptable. The estimates of minimal clinically important change were 11 points and 28% for absolute and relative change scores, respectively. CONCLUSION: The Danish consensus version of the PSS-10 appears to be feasible for use in clinical research settings and has good psychometric properties in terms of agreement, reliability, validity, responsiveness, and interpretability.

Although the importance of humoral immunity to malaria has been established, factors that control antibody production are poorly understood. Follicular helper T cells (Tfh cells) are pivotal for generating high-affinity, long-lived antibody responses. While it has been proposed that expansion of antigen-specific Tfh cells, interleukin (IL) 21 production and robust germinal center formation are associated with protection against malaria in mice, whether Tfh cells are found during Plasmodium vivax (P. vivax) infection and if they play a role during disease remains unknown. Our goal was to define the role of Tfh cells during P. vivax malaria. We demonstrate that P. vivax infection triggers IL-21 production and an increase in Tfh cells (PD-1+ICOS+CXCR5+CD45RO+CD4+CD3+). As expected, FACS-sorted Tfh cells, the primary source of IL-21, induced immunoglobulin production by purified naïve B cells. Furthermore, we found that P. vivax infection alters the B cell compartment and these alterations were dependent on the number of previous infections. First exposure leads to increased proportions of activated and atypical memory B cells and decreased frequencies of classical memory B cells, whereas patients that experienced multiple episodes displayed lower proportions of atypical B cells and higher frequencies of classical memory B cells. Despite the limited sample size, but consistent with the latter finding, the data suggest that patients who had more than five infections harbored more Tfh cells and produce more specific antibodies. P. vivax infection triggers IL-21 production by Tfh that impact B cell responses in humans.

Introduction: The automated counting of reticulocytes has some advantages over the manual method routinely used in clinical laboratories. Technological innovations provide more statistically reliable results, while optimizing the time to perform this test. However, the cost for implementing the automated procedure in laboratory routines still constitutes a barrier to its use in small- and medium-size Brazilian laboratories. Objective: This study evaluated the performance of a new laboratory protocol for reticulocyte counting by flow cytometry using acridine orange (FC/AO), compared with the manual method and with another automated one by flow cytometry using the commercial kit BD Retic-Count (FC/RC) Conclusion: The results showed that, besides being comparable to the manual method, still considered standard, the evaluated new protocol is economically more advantageous than the automated methods currently available, and its cost is comparable to that of the manual method for laboratories that already have appropriate equipment and infrastructure.

Introduction: Currently, the reticulocyte counting is a challenge for clinical laboratories in Brazil, mainly for the ordinary ones, which still use the manual method. This method has some limitations, since it consists of a laborious method, time consuming, with low accuracy. Objectives: This study has developed and evaluated the performance of a New Laboratory Protocol for flow cytometry (FC) reticulocytes counting using acridine orange (AO) as dye, aiming to standardize a more precise, easy, fast implementation, and low cost protocol. After standardization of the New Protocol (FC/AO), it was compared with the manual method. The results were analyzed according to the recommendations of the National Committee for Clinical Laboratory Standards (NCCLS), now known as Clinical and Laboratory Standards Institute (CLSI), to evaluate the interchangeability of methods in linear regression analysis and paired t test, besides other quality control tests. Conclusion: Based on these results concerning to the correlation between the methods and the tests related to quality control, we can admit that FC/AO for reticulocyte counting shows undeniable advantages when compared to the preexisting manual method.

Flavonoids are one of the most important compound groups applied as medicine given their antioxidant properties, but several intrinsic properties can be improved through structural modifications to their molecules. Here, the synthesis and characterization of a new gallium (III) complex with quercetin is described. Electrochemical properties, as well as antioxidant and cytotoxic activities, were investigated and compared to the free flavonoid molecule. The mononuclear complex obtained, [Ga(C15H9O7)3].2H2O.2CH3OH.CH3CH2OH, seems more active as a DPPH radical scavenger given its lower oxidation potential compared to quercetin. The new complex cytotoxic responses have shown to be more effective than those of the free flavonoid and of lapachol used as a control.

O objetivo deste trabalho consistiu em elaborar e avaliar a aceitação de bebida mista de extrato hidrossolúvel de soja (EHS) e suco de amora (SA). As bebidas, produzidas com diferentes proporções de EHS e SA (1:1; 1:1,5; 1:2 m/m), e diferentes concentrações de sólidos solúveis (10, 12 e 14 °Brix), foram analisadas sensorialmente e seus resultados foram submetidos à análise de variância e regressão. As diferentes proporções de EHS e SA nas bebidas produzidas interferiram no aroma e na aparência das bebidas; porém, não existiu uma relação direta entre essas proporções e a aceitabilidade desses atributos. O teor de sólidos solúveis, entretanto, influenciou o sabor e a impressão global. A preferência dos provadores, tanto no sabor como na impressão global, recaiu nas bebidas mais doces, havendo uma relação direta entre o teor de açúcar e a aceitação. Todas as bebidas formuladas foram aprovadas pela equipe de provadores.The purpose of this study was to prepare and evaluate the acceptability of a mixed hydrosoluble soybean extract (EHS) and mulberry juice (SA) drink. The drinks, made with different proportions of EHS and SA (1:1, 1:1.5 and 1:2 w/w) and different concentrations of soluble solids (10, 12 and 14 °Brix), were analyzed by a sensory evaluation and the results submitted to an analysis of variance and regression. The different proportions of EHS and SA interfered in the beverage aroma and appearance, but there was no direct relationship between these proportions and acceptability of the attributes. However, the soluble solids content affected the taste and overall impression. The preference of the panellists, both with respect to flavour and overall impression, was for the sweeter drinks, and there was a direct relationship between the sugar content and acceptance. All the beverages were approved by the panellists.

Resilience can be an important factor in health promotion. The aim of the present study was to carry out a review of the literature in the Pubmed and PsycINFO databases, using the descriptors “resilience” and “chronic disease”. The research contemplated publications conducted in the past 20 years from June 1993 to June 2013. Twelve articles that met the inclusion criteria were identified. These articles pointed towards a negative relationship between resilience and depression, anxiety, incapacitation, and somatization, and also found an inverse correlation between resilience scores and the progression of illness (activity of the disease, control of glycemic level, and severity of depression), and an association between resilience and quality of life and health promotional behavior. In conclusion, resilience may influence the process of illness and outcome in health. It is necessary to develop preventive interventions that allow protective factors for resilience to be developed, which could improve the outcomes in health.

The high mortality rate of candidemia and the limited option for the treatment of Candida spp. infection have been driving the search for new molecules with antifungal property. In this context, coordination complexes of metal ions and ligands appear to be important. Therefore, this study aimed to synthesize two new copper(II) complexes with 2-thiouracil and 6-methyl-2-thiouracil ligands and to evaluate their mutagenic potential and antifungal activity against Candida. The complexes were synthesized and characterized by infrared vibrational spectroscopy, CHN elemental analysis, UV-Vis experiments and ESI-HRMS spectrometry studies. The antifungal activity was evaluated by broth microdilution against 21 clinical isolates of Candida species. The mutagenic potential was evaluated by the Ames test. The complexes were Cu(Bipy)Cl2(thiouracil) (Complex 1) and Cu(Bipy)Cl2(6-methylthiouracil) (Complex 2). Complex 1 showed fungicidal and fungistatic activities against all isolates. Furthermore, the Minimum Inhibitory Concentration (MIC) from 31 to 125 µg/mL and inhibition percentage of 9.9% against the biofilms of C. krusei and C. glabrata were demonstrated. At the concentrations tested, complex 1 exhibited no mutagenic potential. Complex 2 and the free ligands exhibited no antifungal activity at the concentrations evaluated. Since complex 1 presented antifungal activity against all the tested isolates and no mutagenic potential, it could be proposed as a potential new drug for anti-Candida therapy.

The paper characterizes the issues related to compensation of reactive power measures, which protect the smart microgrids from the loss of voltage stability. Slower forms of voltage instability are analyzed with power distribution simulations. The simulations represent the behavior of the system after preset shutdowns; P-U and Q-U charts are drawn to assess the voltage stability reserve in the given time. The purpose of the compensation is to decrease the reactive power transmission and the losses on the smart grid related to this power. This most often translates into introduction of new sources to achieve the established goal. This paper explains the algorithm for optimization of artificial measures of reactive power compensation with the use of decision trees, which are the primary method of induction education of machines due to their high effectiveness and the capability of a simple programming implementation.

Few studies have been done to determine Agaricus brasiliensis Wasser et al. (A. blazei; A. subrufescens) basic mycelial growth characteristics on axenic cultivation. This study aimed to determine the optimal temperature and initial pH for mycelial growth of A. brasiliensis on malt extract agar medium to develop axenic cultivation techniques. Studied initial pH values for mycelial growth were adjusted to 3.0, 4.0, 5.0, 5.5, with HCl, 6.0, 7.0, 8.0, with NaOH, and again 7.0 and 8.0, with CaCO3. Studied temperatures for mycelial growth were 22 ºC, 25 ºC, 28 ºC, 31 ºC and 34 ºC. It was concluded that A. brasiliensis can grow in axenic cultivation at temperature range from 22 oC to 34 ºC, with optimal temperature range from 28 oC to 31 ºC and optimal temperature value of 30.5 ºC ± 0.3 ºC. It also grows in initial pH range from 4.0 to 7.0, adjusted with HCl or NaOH but not CaCO3, with optimal initial pH range from 5.5 to 6.0 and optimal initial pH value of 5.56 ± 0.05. Mycelial growth is inhibited with pH of 3.0 or lower, 8.0 or higher, or when CaCO3 is used to adjust pH in the substratum to 7.0 or higher.Poucos estudos foram desenvolvidos para determinar as condições básicas de crescimento micelial do fungo Agaricus brasiliensis Wasser et al. (A. blazei, A. subrufescens). O objetivo deste trabalho foi determinar a faixa ótima de temperatura e pH para o crescimento micelial, em agar-extrato-de-malte, de A. brasiliensis, visando o desenvolvimento de técnicas de cultivo axênica. Os valores de pH estudados foram 3,0, 4,0, 5,0 e 5,5, ajustados com HCl, 6,0, 7,0 e 8,0, ajustados com NaOH, e 7,0 e 8,0, ajustados com CaCO3. As temperaturas de crescimento estudadas foram 22 ºC; 25 ºC; 28 ºC; 31 ºC e 34 oC. Concluiu-se que A. brasiliensis cresce em uma faixa de temperatura ótima de 28 oC a 31 ºC, com valor ótimo de temperatura de 30,5 ºC ± 0,3 ºC. A faixa de pH inicial ótimo no substrato é de 5,5 a 6,0 e o valor de pH inicial ótimo é de 5,56 ± 0,05. O crescimento do micélio é inibido com pH de 3,0 ou inferior, 8,0 ou superior, ou quando CaCO3 é utilizado para ajuste do pH para 7,0 ou superior.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper. [ABSTRACT FROM AUTHOR]

Background. Recent studies have shown that the inflammatory process, including the biomarker production, and the intense activation of innate immune responses are greater in the malaria caused by Plasmodium vivax than other species. Here, we examined the levels of serum biomarkers and their interaction during acute malaria. Material and Methods. Blood samples were collected from P. vivax-infected patients at admission and from healthy donors. Levels of serum biomarkers were measured by Cytometric Bead Assay or ELISA. Results. P. vivax infection triggered the production of both inflammatory and regulatory biomarkers. Levels of IL-6, CXCL-8, IFN-γ, IL-5, and IL-10 were higher in P. vivax-infected patients than in healthy donors. On the other hand, malaria patients produced lower levels of TNF-α, IL-12p70, and IL-2 than healthy individuals. While the levels of IL-10 and IL-6 were found independent on the number of malaria episodes, higher levels of these cytokines were seen in patients with higher parasite load. Conclusion. A mixed pattern of proinflammatory and regulatory biomarkers is produced in P. vivax malaria. Analysis of biomarker network suggests that IL-10 and IL-6 are a robust axis in malaria patients and that this interaction seems to be associated with the parasite load.

The origin of de novo synthesized proteins in the interface between pea and a Rhizobium sp. in root nodules has been determined. The symbiotic interface is defined as the peribacteroid space including proteins associated with either the symbiosome membrane or the bacteroid outer membrane. Two approaches have been used to study the origin of proteins in the symbiotic interface. First, to determine the localization of de novo synthesized plant-produced proteins in the symbiosomes, an in vitro protein translocation assay was established. To produce plant proteins poly A+ RNA was isolated from root nodules followed by in vitro translation in the presence of [35S]methionine. Subsequently, purified symbiosomes were incubated with the [35S]methionine-labeled plant proteins. The symbiosomes were subfractionated and de novo synthesized plant proteins in the different fractions were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorography. These import studies demonstrate the presence of plant proteins in the symbiosome membrane, peribacteroid space, and bacteroid membrane pellet. In contrast, no proteins of plant origin were detected in the bacteroid cytosol. Second, the presence of de novo synthesized bacteroid proteins in the interface was examined by incubation of symbiosomes isolated under micro-aerobic or aerobic conditions with [35S]methionine. Analysis of symbiosome compartments by SDS-PAGE and phosphor image revealed that proteins of bacteroid origin are primarily detected in the bacteroid cytosol and bacteroid membrane pellet. However, a few bacteroid-produced proteins are also observed in the symbiosome membrane. Together these data demonstrate that the majority of de novo synthesized proteins in the pea-Rhizobium sp. symbiotic interface are of plant origin.

A highly sensitive, automated, purely add-on, high-throughput pseudovirion-based neutralization assay (HT-PBNA) with excellent repeatability and run-to-run reproducibility was developed for human papillomavirus types (HPV) 16, 18, 31, 45, 52, 58 and bovine papillomavirus type 1. Preparation of 384 well assay plates with serially diluted sera and the actual cell-based assay are separated in time, therefore batches of up to one hundred assay plates can be processed sequentially. A mean coefficient of variation (CV) of 13% was obtained for anti-HPV 16 and HPV 18 titers for a standard serum tested in a total of 58 repeats on individual plates in seven independent runs. Natural antibody response was analyzed in 35 sera from patients with HPV 16 DNA positive cervical intraepithelial neoplasia grade 2+ lesions. The new HT-PBNA is based on Gaussia luciferase with increased sensitivity compared to the previously described manual PBNA (manPBNA) based on secreted alkaline phosphatase as reporter. Titers obtained with HT-PBNA were generally higher than titers obtained with the manPBNA. A good linear correlation (R(2) = 0.7) was found between HT-PBNA titers and anti-HPV 16 L1 antibody-levels determined by a Luminex bead-based GST-capture assay for these 35 sera and a Kappa-value of 0.72, with only 3 discordant sera in the low titer range. In addition to natural low titer antibody responses the high sensitivity of the HT-PBNA also allows detection of cross-neutralizing antibodies induced by commercial HPV L1-vaccines and experimental L2-vaccines. When analyzing the WHO international standards for HPV 16 and 18 we determined an analytical sensitivity of 0.864 and 1.105 mIU, respectively.

BACKGROUND:The activation of innate immune responses by Plasmodium vivax results in activation of effector cells and an excessive production of pro-inflammatory cytokines that may culminate in deleterious effects. Here, we examined the activation and function of neutrophils during acute episodes of malaria. MATERIALS AND METHODS:Blood samples were collected from P. vivax-infected patients at admission (day 0) and 30-45 days after treatment with chloroquine and primaquine. Expression of activation markers and cytokine levels produced by highly purified monocytes and neutrophils were measured by the Cytometric Bead Assay. Phagocytic activity, superoxide production, chemotaxis and the presence of G protein-coupled receptor (GRK2) were also evaluated in neutrophils from malaria patients. PRINCIPAL FINDINGS:Both monocytes and neutrophils from P. vivax-infected patients were highly activated. While monocytes were found to be the main source of cytokines in response to TLR ligands, neutrophils showed enhanced phagocytic activity and superoxide production. Interestingly, neutrophils from the malaria patients expressed high levels of GRK2, low levels of CXCR2, and displayed impaired chemotaxis towards IL-8 (CXCL8). CONCLUSION:Activated neutrophils from malaria patients are a poor source of pro-inflammatory cytokines and display reduced chemotactic activity, suggesting a possible mechanism for an enhanced susceptibility to secondary bacterial infection during malaria.

Abstract Background The early peritoneal invasion of epithelial ovarian cancer (EOC) by tumoral aggregates presents in ascites is a major concern. The role of the microenvironment seems to be important in this process but the lack of adequate models to study cellular interactions between cancer cells and stromal cells does not allow to uncover the molecular pathways involved. Our goal was to study the interactions between ovarian cancer cells (OCC) and mesenchymal stem cells (MSC) using a 3D model. Methods We used millimetric pieces of amniochorionic membrane - referred to as amniotic membrane scaffold (AMS) - to create 3D peritoneal nodules mimicking EOC early invasion. We were able to measure the distribution and the depth of infiltration using confocal microsopy. We extracted MSC from the amniochorionic membrane using the markers CD34-, CD45-, CD73+, CD90+, CD105+ and CD29+ at the Fluorescence Activated Cell Sorting (FACS) analysis. We used transwell and wound healing tests to test OCC migration and invasion in vitro. Results Here we show that OCC tumors were located in regions rich in MSC (70%). The tumors infiltrated deeper within AMS in regions rich in MSC (p Conclusions The use of tridimensional models using AMS could be a useful tool to decipher early molecular events in ovarian cancer metastasis. Cytokine inhibitors interrupting the cross-talk between OCCs and MSCs such as IL6 should be investigated as a new therapeutic approach in ovarian cancer.

Resumo Paciente do sexo feminino, 29 anos, ex-tabagista, diagnosticada em setembro de 2012 com doença de Graves e apresentação rápida de exoftalmia bilateral. Na avaliação oftalmológica, apresentava motilidade preservada, proptose e bolsa de gordura superior em AO com retração de PPSS e PPII e exoftalmetria em OD de 26 mm,e em OE de 24 mm. Em maio de 2014,fez o mapeamento da retina que evidenciou cicatrizes de coriorretinite em ambos os olhos e campimetria computadorizada, apresentando degrau nasal em OD, contração superior, depressão centro-inferior. Em junho de 2016, realizou cirurgia de descompressão orbitária de paredes medial e inferior bilateral por via endoscópica com uso de endoscópio nasal Karl Storz, em 30 graus de óptica. A abordagem cirúrgica da oftalmopatia de Graves deve ser empregada na fase cicatricial exceto nos casos com risco de perda da visão. Antes realizada por acesso externo, atualmente a descompressão orbitária pode ser realizada via endoscópica, com mínima invasividade e permite a remoção da parede inferior e medial sem necessidade de incisões externas. É um procedimento seguro para o tratamento da orbitopatia distireoidiana associada a menor morbidade, no qual se evita lesões ao ducto nasolacrimal, nasofrontal ou ao infraorbital e se possibilita redução da proptose entre 3 a 4 mm. Os benefícios da descompressão estão relacionados com a melhora da acuidade visual, além do resultado estético. A continuidade do tratamento cirúrgico será realizada por meio de correção de retração palpebral seguida de blefaroplastia.

Abstract: We investigated the role of different TLRs and MyD88 in host resistance to infection and malaria pathogenesis. TLR2−/−, TLR4−/−, TLR6−/−, TLR9−/− or CD14−/− mice showed no change in phenotypes (parasitemia, body weight and temperature) when infected with Plasmodium chabaudi chabaudi (AS). MyD88−/− mice displayed comparable ability to wild type animals in controlling and clearing parasitemia. Importantly, MyD88−/− mice exhibited impaired production of TNF-α and IFN-γ as well as attenuated symptoms, as indicated by changes in body weight and temperature during parasitemia. Consistently, CD11b+ monocytes and CD11c+ dendritic cells from infected MyD88−/− mice were shown impaired for production of pro-inflammatory cytokines, and in initiating CD4+ T cell responses. Importantly, the inhibition of T cell activation with anti-CD134L, mostly inhibited IFN-γ, partially inhibited TNF-α production, and protected the animals from malaria symptoms. Our findings suggest that MyD88 and possibly its associated TLRs expressed by dendritic cells play an important role in pro-inflammatory responses, T cell activation, and pathogenesis of malaria, but are not critical for the immunological control of the erythrocytic stage of P. chabaudi. [Copyright &y& Elsevier]

Since the introduction of highly active antiretroviral therapies (ART), the prognosis for HIV-1 patients has improved immensely. However, approximately 25% of patients can experience a variety of inflammatory symptoms that are collectively known as immune reconstitution inflammatory syndrome (IRIS). Studying the etiology and immunopathology of IRIS has been hampered by the fact that the symptoms and associated opportunistic infections are highly varied. We hypothesized that there is a common mechanism underlying IRIS pathogenesis and investigated a patient group with IRIS related to different pathogens. Functional and phenotypic characterization of PBMC samples was performed by polychromatic flow cytometry after in vitro stimulation with relevant antigenic preparations. In most patients, IRIS events were characterized by the robust increase of preexisting polyfunctional, highly differentiated effector CD4+ T-cell responses that specifically targeted the antigens of the underlying co-infection. T-cell responses to HIV-1 or other underlying infections were not affected and did not differ between IRIS and non-IRIS patients. These data suggest that patients with IRIS do not have a generalized T-cell dysfunction; instead, IRIS represents a dysregulated CD4+ T-cell response against residual opportunistic infection antigen. These studies were registered at www.clinical-trials.gov as NCT00557570 and NCT00286767. [ABSTRACT FROM AUTHOR]

We have previously shown that mice with loss of C-type lectin-like type II (CLEC2), which have lymphatic dysfunction due to the role of CLEC2 in platelets for maintaining separation between the venous and lymphatic system, develop lung tertiary lymphoid organ (TLO) formation and lung injury that resembles an emphysema phenotype of chronic obstructive pulmonary disease (COPD). We now sought to investigate whether and how TLOs in these mice may play a pathogenic role in lung injury that is relevant to human disease. We found that inhibiting TLO formation using an anti-CD20 antibody in CLEC2-deficient mice partially blocked the development of emphysema. TLOs in CLEC2-deficient mice were rich in plasma cells and were a source of a broad array of autoantibodies. Chronic cigarette smoke exposure increased the size and number of lung TLOs in CLEC2-deficient mice, and was associated with increased markers of antigen presentation and maturation, leading to increased autoantibody deposition. Using lung tissue from COPD patients, we found an increase in lymphatic markers in patients with an emphysema phenotype and autoreactive TLOs compared to COPD patients without emphysema that lack prominent TLOs. Taken together, these results demonstrate that emphysema in mice with lymphatic dysfunction can be partially rescued by blocking TLO formation, and that these TLOs are source of autoantibodies that are exacerbated by cigarette smoke. Our work suggests that lymphatic dysfunction in mice may recapitulate some aspects an autoimmune emphysema phenotype that is seen in a subset of patients with COPD.

Dark fermentation (DF) is a waste treatment bioprocess which produces biohydrogen and volatile fatty acids (VFAs) such as acetate or butyrate. DF can be coupled with microalgae cultivation, allowing VFA conversion into valuable biomass. Nevertheless, the process is hindered by slow butyrate consumption. In this study, novel artificial microalgae-bacteria consortia were used as a strategy to accelerate butyrate removal. Three microalgal strains with various trophic metabolisms, Chlorella sorokiniana, Euglena gracilis and Ochromonas danica, were cultivated on DF effluent that was either sterile or contained endogenous bacteria. Bacteria did not impact microalgal biomass production of C. sorokiniana or E. gracilis while accelerating butyrate removal rates 2 to 10-fold. O. danica greatly impacted microbial diversity, probably due to its phagotrophic metabolism. These results show that bacteria in organic rich effluents can greatly aid in substrate removal while allowing microalgal growth, inspiring bioprocesses coupling raw fermentation effluents with microalgae biomass production and valorization., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The Gleason score is an important predictor of prognosis in prostate cancer. However, its subjective nature can result in over- or under-grading. Our objective was to train an artificial intelligence (AI)-based algorithm to grade prostate cancer in specimens from patients who underwent radical prostatectomy (RP) and to assess the correlation of AI-estimated proportions of different Gleason patterns with biochemical recurrence-free survival (RFS), metastasis-free survival (MFS), and overall survival (OS). Training and validation of algorithms for cancer detection and grading were completed with three large datasets containing a total of 580 whole-mount prostate slides from 191 RP patients at two centers and 6218 annotated needle biopsy slides from the publicly available Prostate Cancer Grading Assessment dataset. A cancer detection model was trained using MobileNetV3 on 0.5 mm × 0.5 mm cancer areas (tiles) captured at 10× magnification. For cancer grading, a Gleason pattern detector was trained on tiles using a ResNet50 convolutional neural network and a selective CutMix training strategy involving a mixture of real and artificial examples. This strategy resulted in improved model generalizability in the test set compared with three different control experiments when evaluated on both needle biopsy slides and whole-mount prostate slides from different centers. In an additional test cohort of RP patients who were clinically followed over 30 years, quantitative Gleason pattern AI estimates achieved concordance indexes of 0.69, 0.72, and 0.64 for predicting RFS, MFS, and OS times, outperforming the control experiments and International Society of Urological Pathology system (ISUP) grading by pathologists. Finally, unsupervised clustering of test RP patient specimens into low-, medium-, and high-risk groups based on AI-estimated proportions of each Gleason pattern resulted in significantly improved RFS and MFS stratification compared with ISUP grading. In summary, deep learning-based quantitative Gleason scoring using a selective CutMix training strategy may improve prognostication after prostate cancer surgery., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

The lymphatic vasculature regulates lung homeostasis through drainage of fluid and trafficking of immune cells and plays a key role in the response to lung injury in several disease states. We have previously shown that lymphatic dysfunction occurs early in the pathogenesis of chronic obstructive pulmonary disease (COPD) caused by cigarette smoke (CS) and that this is associated with increased thrombin and fibrin clots in lung lymph. However, the direct effects of CS and thrombin on lymphatic endothelial cells (LECs) in COPD are not entirely clear. Studies of the blood vasculature have shown that COPD is associated with increased thrombin after CS exposure that causes endothelial dysfunction characterized by changes in the expression of coagulation factors and leukocyte adhesion proteins. Here, we determined whether similar changes occur in LECs. We used an in vitro cell culture system and treated human lung microvascular lymphatic endothelial cells with cigarette smoke extract (CSE) and/or thrombin. We found that CSE treatment led to decreased fibrinolytic activity in LECs, which was associated with increased expression of plasminogen activator inhibitor 1 (PAI-1). LECs treated with both CSE and thrombin together had a decreased expression of tissue factor pathway inhibitor (TFPI) and increased expression of adhesion molecules. RNA sequencing of lung LECs isolated from mice exposed to CS also showed upregulation of prothrombotic and inflammatory pathways at both acute and chronic exposure time points. Analysis of publicly available single-cell RNA sequencing of LECs as well as immunohistochemical staining of lung tissue from COPD patients supported these data and showed increased expression of inflammatory markers in LECs from COPD patients compared to those from controls. These studies suggest that in parallel with blood vessels, the lymphatic endothelium undergoes inflammatory changes associated with CS exposure and increased thrombin in COPD. Further research is needed to unravel the mechanisms by which these changes affect lymphatic function and drive tissue injury in COPD., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Trivedi, Lu, Summers, Kim, Rhee, Houghton, Byers, Lis and Reed.)

Schizophrenia (SCZ) is a complex neurodevelopmental disorder characterized by the manifestation of psychiatric symptoms in early adulthood. While many research avenues into the origins of SCZ during brain development have been explored, the contribution of endothelial/vascular dysfunction to the disease remains largely elusive. To model the neuropathology of SCZ during early critical periods of brain development, we utilized patient-derived induced pluripotent stem cells (iPSCs) to generate 3D cerebral organoids and define cell-specific signatures of disease. Single-cell RNA sequencing revealed that while SCZ organoids were similar in their macromolecular diversity to organoids generated from healthy controls (CTRL), SCZ organoids exhibited a higher percentage of endothelial cells when normalized to total cell numbers. Additionally, when compared to CTRL, differential gene expression analysis revealed a significant enrichment in genes that function in vessel formation, vascular regulation, and inflammatory response in SCZ endothelial cells. In line with these findings, data from 23 donors demonstrated that PECAM1 + microvascular vessel-like structures were increased in length and number in SCZ organoids in comparison to CTRL organoids. Furthermore, we report that patient-derived endothelial cells displayed higher paracellular permeability, implicating elevated vascular activity. Collectively, our data identified altered gene expression patterns, vessel-like structural changes, and enhanced permeability of endothelial cells in patient-derived models of SCZ. Hence, brain microvascular cells could play a role in the etiology of SCZ by modulating the permeability of the developing blood brain barrier (BBB)., (© 2024. The Author(s).)

Abstract: Lis, R, Szymanski, DJ, Qiao, M, and Crotin, RL. An exploratory investigation into the impact of bilateral and unilateral jump characteristics on ground reaction force applications in baseball pitching. J Strength Cond Res 37(9): 1852-1859, 2023-Jump tests are effective, valid, and reliable in examining lower-body power that can influence ground reaction forces (GRFs) in baseball pitching. Relationships between drive and stride leg GRFs and fastball velocities while pitching from the wind-up and stretch were evaluated for performance influences across the following conditions: (a) lower-body GRFs from unilateral countermovement jumps (UCMJ) and bilateral countermovement jumps (BCMJ) and (b) BCMJ and drive and stride leg UCMJ height. Nineteen Division I collegiate baseball pitchers (age; 19.9 ± 1.5 years, height; 1.86 ± 0.06 m, body mass; 90.7 ± 13.8 kg) completed BCMJ and UCMJ tests and threw four-seam fastballs from a pitching mound with 2 embedded force plates. Three moderate associations ( r ≥ 0.47) were statistically significant ( p < 0.05) for BCMJ and UCMJ heights to pitching GRFs. Stride leg UCMJ height was significantly greater than drive leg UCMJ height, p < 0.01, η 2 = 0.34. Wind-up and stretch GRFs were statistically similar. Fastball velocities showed a statistically significant association that was moderately high with wind-up and stretch stride leg anterior-posterior GRFs ( r ≥ 0.65, p < 0.01). Collegiate pitchers jumped significantly higher with their stride leg, and the sum of vertical unilateral jump height of both legs was significantly greater (27%) than BCMJ height to indicate greater single-leg dominance in jumping. Although stride leg height was higher, improving stride leg jumping performance may be functionally more important in producing greater momentum into foot strike that has the potential to increase fastball velocity., (Copyright © 2022 National Strength and Conditioning Association.)

Context: Postoperative hyponatremia leads to prolonged hospital length of stay and readmission within 30 days., Objective: To assess 3 strategies for reducing rates of postoperative hyponatremia and analyze risk factors for hyponatremia., Design: Two retrospective analyses and 1 prospective study., Setting: Tertiary referral hospital., Patients: Patients undergoing transsphenoidal surgery for pituitary adenomas and other sellar and parasellar pathologies., Intervention(s): Phase 1: no intervention. Phase 2: postoperative day (POD) 7 sodium testing and patient education. Phase 3: fluid restriction to 1 L/day on discharge in addition to phase 2 interventions., Main Outcome Measures: Rates of early and delayed hyponatremia and readmissions. Secondary outcomes were risk factors for hyponatremia and readmission costs., Results: In phase 1, 296 patients underwent transsphenoidal surgery. Twenty percent developed early and 28% delayed hyponatremia. Thirty-eight percent underwent POD 7 sodium testing. Readmission rates were 15% overall and 4.3% for hyponatremia. In phase 2 (n = 316), 22% developed early and 25% delayed hyponatremia. Eighty-nine percent complied with POD 7 sodium testing. Readmissions were unchanged although severity of hyponatremia was reduced by 60%. In phase 3 (n = 110), delayed hyponatremia was reduced 2-fold [12.7%, relative risk (RR) = 0.52] and readmissions 3-fold [4.6%, RR = 0.30 (0.12-0.73)]; readmissions for hyponatremia were markedly reduced. Hyponatremia readmission increased costs by 30%., Conclusions: Restricting fluid to 1 L/day on discharge decreases rates of delayed hyponatremia and readmissions by 50%. Standardized patient education and POD 7 sodium testing decreases severity of hyponatremia but does not impact readmission rates. These protocols should be considered standard practice for patients undergoing transsphenoidal surgery., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Endocrine Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The implementation of a sustainable bio-based economy is considered a top priority today. There is no doubt about the necessity to produce renewable bioenergy and bio-sourced chemicals to replace fossil-derived compounds. Under this scenario, strong efforts have been devoted to efficiently use organic waste as feedstock for biohydrogen production via dark fermentation. However, the technoeconomic viability of this process needs to be enhanced by the valorization of the residual streams generated. The use of dark fermentation effluents as low-cost carbon source for microalgae cultivation arises as an innovative approach for bioproducts generation (e.g., biodiesel, bioactive compounds, pigments) that maximizes the carbon recovery. In a biorefinery context, after value-added product extraction, the spent microalgae biomass can be further valorised as feedstock for biohydrogen production. This integrated process would play a key role in the transition towards a circular economy. This review covers recent advances in microalgal cultivation on dark fermentation effluents (DFE). BioH 2 via dark fermentation processes and the involved metabolic pathways are detailed with a special focus on the main aspects affecting the effluent composition. Interesting traits of microalgae and current approaches to solve the challenges associated to the integration of dark fermentation and microalgae cultivation are also discussed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this review., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Volatile fatty acids found in effluents of the dark fermentation of biowastes can be used for mixotrophic growth of microalgae, improving productivity and reducing the cost of the feedstock. Microalgae can use the acetate in the effluents very well, but butyrate is poorly assimilated and can inhibit growth above 1 gC.L -1 . The non-photosynthetic chlorophyte alga Polytomella sp. SAG 198.80 was found to be able to assimilate butyrate fast. To decipher the metabolic pathways implicated in butyrate assimilation, quantitative proteomics study was developed comparing Polytomella sp. cells grown on acetate and butyrate at 1 gC.L -1 . After statistical analysis, a total of 1772 proteins were retained, of which 119 proteins were found to be overaccumulated on butyrate vs. only 46 on acetate, indicating that butyrate assimilation necessitates additional metabolic steps. The data show that butyrate assimilation occurs in the peroxisome via the β-oxidation pathway to produce acetyl-CoA and further tri/dicarboxylic acids in the glyoxylate cycle. Concomitantly, reactive oxygen species defense enzymes as well as the branched amino acid degradation pathway were strongly induced. Although no clear dedicated butyrate transport mechanism could be inferred, several membrane transporters induced on butyrate are identified as potential condidates. Metabolic responses correspond globally to the increased needs for central cofactors NAD, ATP and CoA, especially in the peroxisome and the cytosol., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Lacroux, Atteia, Brugière, Couté, Vallon, Steyer and van Lis.)

Introduction: The emergency department (ED) is at the forefront for treatment of sexual assault patients. Many require treatment for injuries sustained during the assault, ranging from mild to severe. Our objective in this study was to characterize types of injuries associated with sexual assault and identify associated factors., Methods: We reviewed ED charts from an inner-city trauma center and nearby community hospital from 2019-2020 for patients age ≥13 years with a chief complaint of sexual assault. We used descriptive statistics, chi square, and logistic regression to characterize demographics and identify factors associated with trauma., Results: A total of 157 patients met inclusion criteria. The mean age was 27.9 years old (range 13-79 years) and 92.4% were female. Adult patients (age >18 years) comprised 77.5% of assaults vs adolescents (age 13-18 years) at 22.3%. Most patients presented to the trauma center compared to the community hospital (69.4% vs 30.6%). The assailants were reported as 61.2% acquaintance, 22.9% stranger, and 15.9% intimate partner. A forensic rape kit was performed in 92 (58.6%) cases. The patient was intoxicated with alcohol in 39 (24.8%) cases, and 22 (14%) patients reported drug-facilitated assault where an unknown substance was given to them. Alcohol (P = 0.95) and drug-facilitated assault (P = 0.64) did not change the occurrence of injuries. Fifty-seven (36.3%) patients exhibited physical trauma on presentation. Forty-five (28.6%) patients had minor injuries of abrasions, lacerations, or contusions. Major trauma was defined as fracture, brain injury, hemorrhage, strangulation, or injury requiring surgical consultation. There were 12 patients with major trauma consisting of fracture injury or nonfatal strangulation. None of the patients required admission. Sexual assault by an intimate partner (odds ratio [OR] 2.6; 95% CI: 1.1-6.5) and being an adult patient compared to adolescent (OR 3.0; 95% CI, 1.1-7.7) was significantly associated with physical trauma. Sexual assault by an intimate partner was also associated with nonfatal strangulation (OR 4.0; 95% CI, 1.1-15.4)., Conclusion: Physical injuries that resulted from sexual assault were mostly minor and occurred in 36% of rape victims. Intimate partner violence was found to be associated with physical trauma as well as nonfatal strangulation. Overall, this study helps us to understand key factors associated with sexual violence.