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The presently employed synthetic cutting fluids in machining applications lead to skin irritations, hazardous aerosols, ingestion etc., to the machine tool operators. Accomplishing sustainable green machining through bio-cutting fluids, is the key motivating factor behind this work, even though the phenomenon of heat generation is inevitable in the machining process. In the present investigation, Citrullus lanatus extract is chosen as a potential candidate for bio-cutting fluid, since it offers multiple benefits such as increased tool life, cleaner production atmosphere, efficacious heat dissipation, healthy operating conditions, etc. Hybrid LM0-6SiCp-4Grp composite is selected for study since; it has potential applications in marine, aerospace, defence and automotive sectors. Experimentation is carried out with Citrullus lanatus extract as bio-cutting fluid in turning of hybrid LM0-6SiCp-4Grp composite under different processing conditions. The presence of green solution based on Citrullus lanatus extract curtails the rise of temperature significantly during turning operation. Higher magnitude of temperature decrease is observed when the percentage of the concentration of Citrullus lanatus extract is increased, irrespective of depth of turning. Bio-nature and cooling qualities of this green-cutting fluid can contribute its share for green machining and sustainable turning environment. The investigations are further progressed using machine learning algorithms based on the different categories of logical regressions implemented through the Python program. Random forest regression shows better results with 99.8% prediction accuracy compared to other machine learning approaches and the decision tree is constructed through the respective regression model. Decision tree has the ability to handle process parameters systematically and predicts the implications of Citrullus lanatus extract in decreasing the temperature at machine-tool interface during machining operation. This tree structured analysis provides a better inference that delivers flexibility in turning parameters and opens up wider options for operation. Thus, decision tree approach ensures optimized usage of production resources and increases the production capacity through judicial selection of cooling approaches associated with the turning process.

Mesorhizobium phage vB_MloS_Cp1R7A-A1 was isolated from soil planted with chickpea in Saskatchewan. It is dissimilar in sequence and morphology to previously described rhizobiophages. It is a B3 morphotype virus with a distinct prolate capsid and belongs to the tailed phage family Siphoviridae. Its genome has a GC content of 60.3% and 238 predicted genes. Putative functions were predicted for 57 genes, which include 27 tRNA genes with anticodons corresponding to 18 amino acids. This represents the highest number of tRNA genes reported yet in a rhizobiophage. The gene arrangement shows a partially modular organization. Most of the structural genes are found in one module, whereas tRNA genes are in another. Genes for replication, recombination, and nucleotide metabolism form the third module. The arrangement of the replication module resembles the replication module of Enterobacteria phage T5, raising the possibility that it uses a recombination-based replication mechanism, but there is also a suggestion that a T7-like replication mechanism could be used. Phage termini appear to be long direct repeats of just over 12 kb in length. Phylogenetic analysis revealed that Cp1R7A-A1 is more closely related to PhiCbK-like Caulobacter phages and other B3 morphotype phages than to other rhizobiophages sequenced thus far.

Background & aims: Genome-wide association studies in primary biliary cholangitis (PBC) have failed to find X chromosome (chrX) variants associated with the disease. Here, we specifically explore the chrX contribution to PBC, a sexually dimorphic complex autoimmune disease. Methods: We performed a chrX-wide association study, including genotype data from 5 genome-wide association studies (from Italy, United Kingdom, Canada, China, and Japan; 5244 case patients and 11,875 control individuals). Results: Single-marker association analyses found approximately 100 loci displaying P < 5 × 10-4, with the most significant being a signal within the OTUD5 gene (rs3027490; P = 4.80 × 10-6; odds ratio [OR], 1.39; 95% confidence interval [CI], 1.028-1.88; Japanese cohort). Although the transethnic meta-analysis evidenced only a suggestive signal (rs2239452, mapping within the PIM2 gene; OR, 1.17; 95% CI, 1.09-1.26; P = 9.93 × 10-8), the population-specific meta-analysis showed a genome-wide significant locus in East Asian individuals pointing to the same region (rs7059064, mapping within the GRIPAP1 gene; P = 6.2 × 10-9; OR, 1.33; 95% CI, 1.21-1.46). Indeed, rs7059064 tags a unique linkage disequilibrium block including 7 genes: TIMM17B, PQBP1, PIM2, SLC35A2, OTUD5, KCND1, and GRIPAP1, as well as a superenhancer (GH0XJ048933 within OTUD5) targeting all these genes. GH0XJ048933 is also predicted to target FOXP3, the main T-regulatory cell lineage specification factor. Consistently, OTUD5 and FOXP3 RNA levels were up-regulated in PBC case patients (1.75- and 1.64-fold, respectively). Conclusions: This work represents the first comprehensive study, to our knowledge, of the chrX contribution to the genetics of an autoimmune liver disease and shows a novel PBC-related genome-wide significant locus.

Microbes are critical drivers of all ecosystems and many biogeochemical processes, yet little is known about how the three-dimensional (3D) organization of these dynamic organisms contributes to their overall function. To probe how biofilm structure affects microbial activity, we developed a technique for patterning microbes in 3D geometries using projection stereolithography to bioprint microbes within hydrogel architectures. Bacteria were printed and monitored for biomass accumulation, demonstrating postprint viability of cells using this technique. We verified our ability to integrate biological and geometric complexity by fabricating a printed biofilm with two E. coli strains expressing different fluorescence. Finally, we examined the target application of microbial absorption of metal ions to investigate geometric effects on both the metal sequestration efficiency and the uranium sensing capability of patterned engineered Caulobacter crescentus strains. This work represents the first demonstration of the stereolithographic printing of microbials and presents opportunities for future work of engineered biofilms and other complex 3D structured cultures.

The nitrogen-fixing symbiosis fixation between legumes and Rhizobium helps the plant to survive and to compete effectively on nitrogen poor soils. The soil environment attached to the root system is characterized by intense microbial activity. This work aimed to study the diversity of indigenous rhizobia living in the rhizosphere of forage pea in eastern and central Algeria. Bacteria were trapped in vitro from collected soils. They were isolated from nodules by direct isolation or by bacterial trapping, and tested for nodulation. Further characterization was carried out to identify the isolates. Ninety-four isolates were obtained. Thirty-four were rhizobial strains that could, with two exceptions, establish root-nodule symbioses with the host plant. Different profiles of susceptibility to rhizobiophages indicated that the majority of isolates were likely to be members of the Rhizobium leguminosarum group of species. The strains revealed diverse plasmid profiles, and diverse restriction patterns of cpn60 gene amplicons and on Southern blot analysis of nodABC genes. Sequences of 16S rRNA gene showed that all isolates exhibited 99.85% identity with R. leguminosarum sv. viciae USDA 2370T, R. laguerreae FB206T and R. anhuiense CCBAU 23252T. Analysis of cpn60 sequences revealed two groups. One of them clustered with ≤99% identity with R. laguerreae FB403. The second was clustered with ≤99% identity with R. laguerreae FB206T. Our findings suggest that the isolates may belong to the species R. laguerreae.

RNA interference (RNAi) is mediated by an ∼21-nt double-stranded small interfering RNA (siRNA) and shows great promise in delineating gene functions and in developing therapeutics for human diseases. However, effective gene silencing usually requires the delivery of multiple siRNAs for a given gene, which is often technically challenging and time-consuming. In this study, by exploiting the type IIS restriction endonuclease-based synthetic biology methodology, we developed the fast assembly of multiplex siRNAs (FAMSi) system. In our proof-of-concept experiments, we demonstrated that multiple fragments containing three, four, or five siRNA sites targeting common Smad4 and/or BMPR-specific Smad1, Smad5, and Smad8 required for BMP9 signaling could be assembled efficiently. The constructed multiplex siRNAs effectively knocked down the expression of Smad4 and/or Smad1, Smad5, and Smad8 in mesenchymal stem cells (MSCs), and they inhibited all aspects of BMP9-induced osteogenic differentiation in bone marrow MSCs (BMSCs), including decreased expression of osteogenic regulators/markers, reduced osteogenic marker alkaline phosphatase (ALP) activity, and diminished in vitro matrix mineralization and in vivo ectopic bone formation. Collectively, we demonstrate that the engineered FAMSi system provides a fast-track platform for assembling multiplexed siRNAs in a single vector, and thus it may be a valuable tool to study gene functions or to develop novel siRNA-based therapeutics., Graphical Abstract, RNA interference shows great promise in delineating gene functions and in developing therapeutics for human diseases. He et al. exploited the type IIS restriction endonuclease-based synthetic biology and developed the fast assembly of multiplex siRNAs (FAMSi) system, which streamlines RNAi-based gene function studies and the development of siRNA-based therapeutics.

There is conflicting data regarding the association of pre-transplant AT1R antibody levels and long-term outcomes following kidney transplantation.We examined the association between pre-transplant antibodies and long-term graft outcome by assaying pre-transplant sera from 125 kidney transplant recipients from 1999 to 2009.The mean age at transplant was 55.7 ± 13 years; 67.2% were male, 87.2% were Caucasian, and 67.2% received a deceased donor transplant. Induction therapy included 44.8% thymoglobulin. Human leukocyte antigen (HLA) donor-specific antibodies (DSA) were present in 22 (17.6%) patients, while AT1R antibodies 17 U/mL were present in 24 (19.2%). The mean AT1R antibodies level was 13 ± 7.2 U/mL. Patients were followed-up for 7.1 ± 1.9 years after transplant. Pre-transplant AT1R antibodies were associated with rejection (p 0.0001), antibody-mediated rejection (ABMR) (p 0.0001), and death-censored graft failure (DCGF) (p = 0.01). This was confirmed by univariate Cox regression analyses for AT1R antibodies 10 U/mL (HR 2.64, 95% Cl 1.35 - 5.17, p = 0.04) and AT1R antibodies 17 U/mL (HR = 1.74, 95% Cl 1.061 - 2.98, p = 0.04). Multivariable analyses did not retain AT1R antibodies as independent predictors of DCGF; however, pre-transplant HLA, DSA, and acute rejection during the first year were associated with DCGF (HR 2.07, 95% Cl 1.13 - 3.78, p = 0.02 and HR 3.03, 95% Cl 1.13 - 3.78, p = 0.0002, respectively).Our study indicates that in patients with a functioning kidney allograft 5 years, pre-transplant AT1R antibodies may be associated with a greater risk of rejection and late graft failure.

In this study, we present the performance as an inhibitor of plant extract of Aerva lanata flowers (ALF) used as an environmentally benign compound for protection of corrosion of low-carbon steel in 1.0 mol L−1 HCl solution. The investigation was accomplished using conventional mass loss, electrochemical measurements and scanning electron microscopy (SEM). The investigation of weight loss demonstrated that the inhibition performance had been improved via raising the concentration of the inhibitor. The efficiency of inhibition has been found to exceed 88% at 600 ppm of the ALF formulation. Using anodic polarization curves, it was demonstrated that flower extract of Aerva lanata acted as a mixed-type inhibitor. The green inhibitor compounds covered the steel surface following Langmuir adsorption isotherm. The surface morphology analysis was conducted to validate the results.

In the developing peripheral nervous system, Schwann cells (SCs) extend their processes to contact, sort, and myelinate axons. The mechanisms that contribute to the interaction between SCs and axons are just beginning to be elucidated. Using a SC-neuron coculture system, we demonstrate that Arg-Gly-Asp (RGD) peptides that inhibit αV -containing integrins delay the extension of SCs elongating on axons. αV integrins in SC localize to sites of contact with axons and are expressed early in development during radial sorting and myelination. Short interfering RNA-mediated knockdown of the αV integrin subunit also delays SC extension along axons in vitro, suggesting that αV -containing integrins participate in axo-glial interactions. However, mice lacking the αV subunit in SCs, alone or in combination with the potentially compensating α5 subunit, or the αV partners β3 or β8 , myelinate normally during development and remyelinate normally after nerve crush, indicating that overlapping or compensatory mechanisms may hide the in vivo role of RGD-binding integrins.

Mesenchymal stem cells (MSCs) are multipotent progenitors that have the ability to differentiate into multiple lineages, including bone, cartilage, and fat. We previously demonstrated that the least known bone morphogenetic protein (BMP)9 (also known as growth differentiation factor 2) is one of the potent osteogenic factors that can induce both osteogenic and adipogenic differentiation of MSCs. Nonetheless, the molecular mechanism underlying BMP9 action remains to be fully understood. Leptin is an adipocyte-derived hormone in direct proportion to the amount of body fat, and exerts pleiotropic functions, such as regulating energy metabolism, bone mass, and mineral density. In this study, we investigate the potential effect of leptin signaling on BMP9-induced osteogenic differentiation of MSCs. We found that exogenous leptin potentiated BMP9-induced osteogenic differentiation of MSCs both in vitro and in vivo, while inhibiting BMP9-induced adipogenic differentiation. BMP9 was shown to induce the expression of leptin and leptin receptor in MSCs, while exogenous leptin upregulated BMP9 expression in less differentiated MSCs. Mechanistically, we demonstrated that a blockade of JAK signaling effectively blunted leptin-potentiated osteogenic differentiation induced by BMP9. Taken together, our results strongly suggest that leptin may potentiate BMP9-induced osteogenesis by cross-regulating BMP9 signaling through the JAK/STAT signaling pathway in MSCs. Thus, it is conceivable that a combined use of BMP9 and leptin may be explored as a novel approach to enhancing efficacious bone regeneration and fracture healing.

Rhizobium leguminosarum strain VF39 contains a plasmid, pRleVF39b, which encodes a distinctive type of conjugation system (rhizobial type IVa) that is relatively widespread among rhizobial genomes. The cluster of genes encoding the transfer functions lacks orthologs to genes such as traCD, traF and traB, but contains 15 conserved genes of unknown function. We determined the importance of these genes in conjugation by constructing marked and unmarked mutations in each gene, and established that six genes, now designated trcA-F, played a significant role in plasmid transfer. Like the relaxase gene, traA, and the genes encoding the MPF system (trb genes), five of these genes, located in two divergently transcribed operons, are regulated by the Xre family repressor TrbR. The other gene, trcF encodes a protein with similarity to histidinol phosphatases, and its role in conjugation is unclear, but mutations in trcF are severely impaired for conjugation. TrcF does not play a role in regulation of other conjugation genes.

IQGAP1 is a scaffold protein involved in a range of cellular activities, including migration, invasion, adhesion and proliferation. It is also oncogenic in a variety of cancers, promoting primary tumor growth and invasiveness. However, the role of IQGAP1 in tumor progression and metastasis remains unclear. In this study, we use both knockdown and knockout of IQGAP1 to investigate its role in the metastatic cascade of both melanoma and breast cancer cells in vivo. We find that reduction of IQGAP1 expression decreases the formation of both spontaneous and experimental metastases, without limiting primary or metastatic tumor growth. Furthermore, IQGAP1 knockout significantly inhibits extravasation of tumor cells from circulation, possibly involving invadopodial function. By expressing mutant forms of IQGAP1 in a knockout context, we also determine that IQGAP1’s pro-metastatic functions are dependent on multiple domains and functions. These data demonstrate that IQGAP1 is crucial for metastasis in vivo through regulation of extravasation and suggest that it may represent a valid therapeutic target for inhibiting metastasis.

The extracellular matrix (ECM) is a complex meshwork of insoluble fibrillar proteins and signaling factors interacting together to provide architectural and instructional cues to the surrounding cells. Alterations in ECM organization or composition and excessive ECM deposition have been observed in diseases such as fibrosis, cardiovascular diseases, and cancer. We provide here optimized protocols to solubilize ECM proteins from normal or tumor tissues, digest the proteins into peptides, analyze ECM peptides by mass spectrometry, and interpret the mass spectrometric data. In addition, we present here two novel R-script-based web tools allowing rapid annotation and relative quantification of ECM proteins, peptides, and intensity/abundance in mass spectrometric data output files. We illustrate this protocol with ECMs obtained from two pairs of tissues, which differ in ECM content and cellularity: triple-negative breast cancer and adjacent mammary tissue, and omental metastasis from high-grade serous ovarian cancer and normal omentum. The complete proteomics data set generated in this study has been deposited to the public repository ProteomeXchange with the data set identifier: PXD005554.

In the stem-cell niche, the extracellular matrix (ECM) serves as a structural support that additionally provides stem cells with signals that contribute to the regulation of stem-cell function, via reciprocal interactions between cells and components of the ECM. Recently, cell-derived ECMs have emerged as in vitro cell culture substrates to better recapitulate the native stem-cell microenvironment outside the body. Significant changes in cell number, morphology and function have been observed when mesenchymal stem cells (MSC) were cultured on ECM substrates as compared to standard tissue-culture polystyrene (TCPS). As select ECM components are known to regulate specific stem-cell functions, a robust characterization of cell-derived ECM proteomic composition is critical to better comprehend the role of the ECM in directing cellular processes. Here, we characterized and compared the protein composition of ECM produced in vitro by bone marrow-derived MSC, adipose-derived MSC and neonatal fibroblasts from different donors, employing quantitative proteomic methods. Each cell-derived ECM displayed a specific and unique matrisome signature, yet they all shared a common set of proteins. We evaluated the biological response of cells cultured on the different matrices and compared them to cells on standard TCPS. The matrices lead to differential survival and gene-expression profiles among the cell types and as compared to TCPS, indicating that the cell-derived ECMs influence each cell type in a different manner. This general approach to understanding the protein composition of different tissue-specific and cell-derived ECM will inform the rational design of defined systems and biomaterials that recapitulate critical ECM signals for stem-cell culture and tissue engineering., National Cancer Institute (Grants U54-CA126515 and U54-CA163109)

Rhizobia are one of the most important and best studied groups of bacterial symbionts. They are defined by their ability to establish nitrogen-fixing intracellular infections within plant hosts. One surprising feature of this symbiosis is that the bacterial genes required for this complex trait are not fixed within the chromosome, but are encoded on mobile genetic elements (MGEs), namely plasmids or integrative and conjugative elements. Evidence suggests that many of these elements are actively mobilizing within rhizobial populations, suggesting that regular symbiosis gene transfer is part of the ecology of rhizobial symbionts. At first glance, this is counterintuitive. The symbiosis trait is highly complex, multipartite and tightly coevolved with the legume hosts, while transfer of genes can be costly and disrupt coadaptation between the chromosome and the symbiosis genes. However, horizontal gene transfer is a process driven not only by the interests of the host bacterium, but also, and perhaps predominantly, by the interests of the MGEs that facilitate it. Thus understanding the role of horizontal gene transfer in the rhizobium–legume symbiosis requires a ‘mobile genetic element's-eye view' on the ecology and evolution of this important symbiosis. This article is part of the theme issue ‘The secret lives of microbial mobile genetic elements’.

Simple Summary In breast cancer development, the expression of Ki-67 is strongly associated with cancer proliferation and is a known indicator of prognosis and outcome. Ki-67 expression levels are also useful to inform treatment decision making in some cases. As a result, routine measurement of Ki-67 is now widely performed during pathological tumour evaluation. However, the Ki-67 appraisal is not without its limitations and shortcomings—the aim of this study was to provide an overview of Ki-67 use in the clinical setting, the current challenges associated with its measurement, and the novel strategies that will hopefully enhance Ki-67 proliferation indices for prospective breast cancer patients. Abstract The advent of molecular medicine has transformed breast cancer management. Breast cancer is now recognised as a heterogenous disease with varied morphology, molecular features, tumour behaviour, and response to therapeutic strategies. These parameters are underpinned by a combination of genomic and immunohistochemical tumour factors, with estrogen receptor (ER) status, progesterone receptor (PgR) status, human epidermal growth factor receptor-2 (HER2) status, Ki-67 proliferation indices, and multigene panels all playing a contributive role in the substratification, prognostication and personalization of treatment modalities for each case. The expression of Ki-67 is strongly linked to tumour cell proliferation and growth and is routinely evaluated as a proliferation marker. This review will discuss the clinical utility, current pitfalls, and promising strategies to augment Ki-67 proliferation indices in future breast oncology.

The rise in harms associated with misuse of substances such as cannabis, alcohol and opioids is a public health issue gaining increasing importance in Canada. Taking a closer look at who is being hospitalized, and for which substances, helps inform efforts to improve access to services for youth. Between 2017 and 2018, hospitalizations for harm caused by substance use accounted for about one in 20 of all hospital stays among youths aged 10-24 years in Canada. Cannabis use was documented in nearly 40% of these hospitalizations, while alcohol was associated with 26%. Approximately one in every six youths (17%), who were hospitalized for harm caused by substance use, was hospitalized more than once for substance use within the same year.

SeaFlow is an underway flow cytometer that provides continuous shipboard observations of the abundance and optical properties of small phytoplankton (, Measurement(s)planktonic material • temperature of water • salinity • light irradiance • plankton size distributionTechnology Type(s)flow cytometer • Temperature Sensor Device • salinometer • photometerFactor Type(s)time • locationSample Characteristic - OrganismCyanobacteria • Micromonas pusilla • Navicula transitans • Thalassiosira pseudonana • Thalassiosira weissflogii • Geminifera cryophila • Crocosphaera • Synechococcus sp. WH 8012 • Synechococcus sp. WH 6501 • Prochlorococcus eMED4 • Prochlorococcus eMIT9312 • Prochlorococcus AS9601 • Prochlorococcus 1314Sample Characteristic - Environmentoceanic sea surface microlayer biome • sea waterSample Characteristic - LocationNorth Pacific Ocean Machine-accessible metadata file describing the reported data: 10.6084/m9.figshare.10110755

Although bone morphogenetic proteins (BMPs) initially showed effective induction of ectopic bone growth in muscle, it has since been determined that these proteins, as members of the TGF-β superfamily, play a diverse and critical array of biological roles. These roles include regulating skeletal and bone formation, angiogenesis, and development and homeostasis of multiple organ systems. Disruptions of the members of the TGF-β/BMP superfamily result in severe skeletal and extra-skeletal irregularities, suggesting high therapeutic potential from understanding this family of BMP proteins. Although it was once one of the least characterized BMPs, BMP9 has revealed itself to have the highest osteogenic potential across numerous experiments both in vitro and in vivo, with recent studies suggesting that the exceptional potency of BMP9 may result from unique signaling pathways that differentiate it from other BMPs. The effectiveness of BMP9 in inducing bone formation was recently revealed in promising experiments that demonstrated efficacy in the repair of critical sized cranial defects as well as compatibility with bone-inducing bio-implants, revealing the great translational promise of BMP9. Furthermore, emerging evidence indicates that, besides its osteogenic activity, BMP9 exerts a broad range of biological functions, including stem cell differentiation, angiogenesis, neurogenesis, tumorigenesis, and metabolism. This review aims to summarize our current understanding of BMP9 across biology and the body.

Anthropogenic acidification in SW‐Scotland, from the early 19th Century onwards, led to the extinction of several loch (lake) brown trout (Salmo trutta) populations and substantial reductions in numbers in many others. Higher altitude populations with no stocking influence, which are isolated above natural and artificial barriers and subjected to the greatest effect of acidification, exhibited the least intrapopulation genetic diversity (34% of the allelic richness of the populations accessible to anadromous S. trutta). These, however, were characterised by the greatest interpopulation divergence (highest pairwise D EST 0.61 and F ST 0.53 in contemporary samples) based on 16 microsatellite loci and are among the most differentiated S. trutta populations in NW‐Europe. Five lochs above impassable waterfalls, where S. trutta were thought to be extinct, are documented as having been stocked in the late 1980s or 1990s. All five lochs now support self‐sustaining S. trutta populations; three as a direct result of restoration stocking and two adjoining lochs largely arising from a small remnant wild population in one, but with some stocking input. The genetically unique Loch Grannoch S. trutta, which has been shown to have a heritable increased tolerance to acid conditions, was successfully used as a donor stock to restore populations in two acidic lochs. Loch Fleet S. trutta, which were re‐established from four separate donor sources in the late 1980s, showed differential contribution from these ancestors and a higher genetic diversity than all 17 natural loch populations examined in the area. Genetically distinct inlet and outlet spawning S. trutta populations were found in this loch. Three genetically distinct sympatric populations of S. trutta were identified in Loch Grannoch, most likely representing recruitment from the three main spawning rivers. A distinct genetic signature of Loch Leven S. trutta, the progenitor of many Scottish farm strains, facilitated detection of stocking with these strains. One artificially created loch was shown to have a population genetically very similar to Loch Leven S. trutta. In spite of recorded historical supplemental stocking with Loch Leven derived farm strains, much of the indigenous S. trutta genetic diversity in the area remains intact, aside from the effects of acidification induced bottlenecks. Overall genetic diversity and extant populations have been increased by allochthonous stocking.

The aim of this work was to evaluate the impact of two factors on the ripening profiles of hard cooked cheeses: (F1) the growth medium for the primary and adjunct cultures, constituted by autochthonous strains: Lactobacillus helveticus 209 (Lh209) and Lactobacillus paracasei 90 (Lp90), respectively, and (F2) the addition of L. paracasei Lp90 as adjunct culture. Four types of cheeses were made: W and M cheeses in which only Lh209 was added after its growth in whey and MRS, respectively; Wa and Ma cheeses in which both strains (Lh209 and Lp90) were added after their growth in whey and MRS, respectively. Physicochemical and microbial composition, proteolysis and profiles of organic acids and volatile compounds were analyzed. According to the methodology of the cultures preparation, W and Wa cheeses showed a higher level of secondary proteolysis and lower level of primary proteolysis (P < 0·05), lower content of citric and acetic acids and higher amount of propionic acid (P < 0·05), in comparison with M and Ma cheeses. The incorporation of Lp90 increased the secondary proteolysis (P < 0·05), decreased the citric acid (P < 0·05), and increased the propionic acid only when was added after their growth in whey (P < 0·05). Both factors significantly modified the percentages of the volatile compounds grouped in chemical families; in addition, for the half of the compounds detected, significant differences were found. Based on the obtained results, the use of Lp90 as an adjunct in hard cooked cheeses, and the preincubation of the cultures in whey are strategies to accelerate the cheese ripening and to enhance the production of some characteristic compounds of this type of cheeses, such as propan-2-one, hexan-2-one, 2- A nd 3-methyl butanal, heptan-2-ol, acetic and 3-methylbutanoic acids and 3-hydroxy butan-2-one. Fil: Cuffia, Facundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; Argentina Fil: Bergamini, Carina Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; Argentina Fil: Wolf, Irma Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; Argentina Fil: Hynes, Erica Rut. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; Argentina Fil: Perotti, Maria Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; Argentina

White-tailed deer Odocoileus virginianus are the most popular big game animal in the United States. Recreational harvest of these animals is a critical tool in population management, as well as an important financial resource for state economies and wildlife agencies. Thus, herd health evaluations can provide information to wildlife managers tasked with developing sustainable harvest practices while monitoring for emergent problems. The purpose of our study was to document causes of illness and natural mortality in New York white-tailed deer submitted for post mortem evaluation. Animals were presented by members of the public and wildlife management personnel due to abnormal behavior or unexplained death. We describe demographic and seasonal associations among gross and histologic evaluation and diagnostic testing. Post mortem examinations were performed on 735 white-tailed deer submitted for necropsy in New York from January 2011 to November 2017. Causes of euthanasia or mortality were classified into nine categories. The most common findings were bacterial infections, trauma not evident at time of collection, and nutritional issues, primarily starvation. Using a multinomial logistic regression model, we looked for associations between the mortality categories and age, sex and season. Compared to the baseline of bacterial deaths, adults were less likely to have died from nutritional and parasitic causes, males were less likely to have died from other causes, and risk of death from nutritional reasons decreased from season to season, with lowest risk in winter. These methods can help wildlife biologists track changes in disease dynamics over time.

Immunosuppressive tumour microenvironments (TME) reduce the effectiveness of immune responses in cancer. Non-haematopoietic mesenchymal stromal cells (MSC), the precursor to cancer associated fibroblasts (CAFs), dictate tumour progression by enhancing immune cell suppression. Hyper-sialylation of glycans promotes immune evasion in cancer, but the role of sialyation in stromal cell-mediated immunosuppression is unknown.Here we study changes in sialyltransferase (ST) enzymes and associated surface expressed sialic acid in stromal cells following inflammatory and tumour secretome conditioning. We show that tumour conditioned stromal cells have increased levels of sialyltransferases, α2,3/6 linked sialic acid and siglec ligands. In tumour models of solid (colorectal cancer) and haematological (multiple myeloma) stromal rich tumours, stromal cell sialylation is associated with enhanced immunosuppression. Using datasets and patient samples, we confirm that targeting sialylation in tumour stromal cells reverses immune cell exhaustion. Targeting stromal cell sialylation may represent a novel immune checkpoint to reactivate anti-tumour immunity.

1.AbstractMesoscale eddies have been shown to support elevated dinitrogen (N2) fixation rates (NFRs) and abundances of N2-fixing microorganisms (diazotrophs), but the mechanisms underlying these observations are not well understood. We explored relationships among NFRs and cyanobacterial diazotroph abundances in eddy pairs of opposite polarity sampled in the North Pacific Subtropical Gyre and compared our observations to seasonal trends from the Hawaii Ocean Time-series (HOT) program. Consistent with previous reports, we found that NFRs were anomalously high for this region (up to 3.7-fold above previous monthly HOT observations) in the centers of the sampled anticyclones, coinciding with elevated abundances of Crocosphaera in the summertime. We then coupled our field-based observations, together with transcriptomic analyses of nutrient stress marker genes and ecological models, to evaluate potential mechanisms controlling diazotroph abundance and activity at the mesoscale. Specifically, we evaluated the role of biological (via estimates of growth and grazing rates) and physical controls on populations of Crocosphaera, Trichodesmium, and diatom symbionts. Our results suggest that increased Crocosphaera abundances associated with summertime anticyclones resulted from the alleviation of phosphate limitation, allowing cells to grow at rates exceeding grazing losses. In contrast, distributions of larger, buoyant taxa (Trichodesmium and diatom symbionts) appeared less affected by eddy-driven biological controls. Instead, they appeared driven by physical dynamics along frontal boundaries that separate cyclonic and anticyclonic eddies. Together, the interplay of eddy-specific changes in bottom-up control, top-down control, and the physical accumulation of cells likely explains the elevated diazotroph abundances and NFRs associated with anticyclones and eddy fronts.

Pneumocystis jirovecii can cause severe pneumonia in immunocompromised patients, which can be life threatening if left untreated. Despite the widespread use of polymerase chain reaction (PCR) within the clinical laboratory setting, FDA-approved PCR assays are not readily available for the detection of Pneumocystis from respiratory samples. Using the Luminex ARIES system-an open-channel, automated, sample-to-answer PCR platform-the cell division cycle 2 (cdc-2) gene can be targeted for the detection of Pneumocystis. This novel TaqMan-based, real-time PCR assay offers improved sensitivity compared to staining or immunofluorescence while reducing turnaround time and eliminating the challenges surrounding microscopic identification. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Primer/probe master mix preparation Basic Protocol 2: Positive control (cdc-2) plasmid preparation Basic Protocol 3: Mucus digestion Basic Protocol 4: Cell lysis Basic Protocol 5: Carrier RNA/proteinase K preparation Basic Protocol 6: Cassette assembly Basic Protocol 7: Running the assay Basic Protocol 8: Interpreting results.

Many bacteria use the second messenger cyclic diguanylate (c-di-GMP) to control motility, biofilm production and virulence. Here, we identify a thermosensory diguanylate cyclase (TdcA) that modulates temperature-dependent motility, biofilm development and virulence in the opportunistic pathogen Pseudomonas aeruginosa. TdcA synthesizes c-di-GMP with catalytic rates that increase more than a hundred-fold over a ten-degree Celsius change. Analyses using protein chimeras indicate that heat-sensing is mediated by a thermosensitive Per-Arnt-SIM (PAS) domain. TdcA homologs are widespread in sequence databases, and a distantly related, heterologously expressed homolog from the Betaproteobacteria order Gallionellales also displayed thermosensitive diguanylate cyclase activity. We propose, therefore, that thermotransduction is a conserved function of c-di-GMP signaling networks, and that thermosensitive catalysis of a second messenger constitutes a mechanism for thermal sensing in bacteria., Many bacteria use the second messenger cyclic diguanylate (c-di-GMP) to control motility, biofilm production and virulence. Here, the authors identify a thermosensitive enzyme that synthesizes c-di-GMP and modulates temperature-dependent motility, biofilm development and virulence in the opportunistic pathogen Pseudomonas aeruginosa.

Highlights • A non-human primate protective head cap that promotes wound healing after cranial implants. • Use of the head cap reduced wound dehiscence and the need to re-suture surgical wounds. • The head cap is easily adjustable to cover most primate cranial implants. • The head cap facilitates primate cranial implant wound management in neuroscience., Background Neuroscience studies with macaque monkeys may require cranial implants to stabilize the head or gain access to the brain for scientific purposes. Wound management that promotes healing after the cranial implant surgery in non-human primates can be difficult as it is not necessarily possible to cover the wound margins. New method Here, we developed an easily modifiable head cap that protects the sutured skin margins after cranial implant surgery and contributes to wound healing. The protective head cap was developed in response to monkeys picking at sutured skin margins around an implant, complicating healing. The user-friendly protective cap, made from Klarity- R™ Sheet (3.2 mm thick with 36 % or 42 % perforation) is affixed to the implant post-surgically. Once secured and while the monkey is still anesthetized, the plastic sheeting is molded around the implant. The protective head cap restricts the monkey’s finger access to its’ wound margins while allowing air to circulate to promote wound healing. Results and comparison with existing methods Across two UK primate facilities, the protective head cap promoted wound healing. In monkeys that did not wear the head cap, re-suturing was necessary in ∼30 % of cases. In contrast, none of the monkeys that wore the head cap required re-suturing. The monkeys wearing the head cap also had reduced numbers of days of prescribed antibiotics and analgesia. Conclusion This bespoken, easily adaptable, protective head cap supports postoperative wound healing, and enhances the welfare of monkeys involved in neuroscience research.

Fibronectin containing alternatively spliced EIIIA and EIIIB domains is largely absent from mature quiescent vessels in adults, but is highly expressed around blood vessels during developmental and pathological angiogenesis. The precise functions of fibronectin and its splice variants during developmental angiogenesis however remain unclear due to the presence of cardiac, somitic, mesodermal and neural defects in existing global fibronectin KO mouse models. Using a rare family of surviving EIIIA EIIIB double KO mice, as well as inducible endothelial-specific fibronectin-deficient mutant mice, we show that vascular development in the neonatal retina is regulated in an autocrine manner by endothelium-derived fibronectin, and requires both EIIIA and EIIIB domains and the RGD-binding α5 and αv integrins for its function. Exogenous sources of fibronectin do not fully substitute for the autocrine function of endothelial fibronectin, demonstrating that fibronectins from different sources contribute differentially to specific aspects of angiogenesis. Electronic supplementary material The online version of this article (doi:10.1007/s10456-017-9563-8) contains supplementary material, which is available to authorized users.

Gambling and substance use disorders are highly comorbid. Both clinical populations are impulsive and exhibit risky decision-making. Drug-associated cues have long been known to facilitate habitual drug-seeking, and the salient audiovisual cues embedded within modern gambling products may like-wise encourage problem gambling. The dopamine neurons of the ventral tegmental area (VTA) are exquisitely sensitive to drugs of abuse, uncertain rewards, and reward-paired cues, and may therefore be the common neural substrate mediating synergistic features of both disorders. To test this hypothesis, we first gained specific inhibitory control over VTA dopamine neurons by transducing a floxed inhibitory DREADD (AAV5-hSyn-DIO-hM4D(Gi)-mCherry) in rats expressing Cre recombinase in tyrosine hydroxylase neurons. We then trained rats in our cued rat gambling task (crGT), inhibiting dopamine neurons throughout task acquisition and performance, before allowing them to self-administer cocaine in the same diurnal period as crGT sessions. The trajectories of addiction differ in women and men, and the dopamine system may differ functionally across the sexes, therefore we used male and female rats here. We found that inhibition of VTA dopamine neurons decreased cue-induced risky choice and reduced motor impulsivity in males, but surprisingly, enhanced risky decision making in females. Inhibiting VTA dopamine neurons also prevented cocaine-induced deficits in decision making in both sexes, but nevertheless drove all animals to consume more cocaine. These findings show that chronic dampening of dopamine signalling can have both protective and deleterious effects on addiction-relevant behaviours, depending on biological sex and dependent variable of interest.

The contribution of biological nitrogen fixation to the total N requirement of food and feed crops diminished in importance with the advent of synthetic N fertilizers, which fueled the “green revolution”. Despite being environmentally unfriendly, the synthetic versions gained prominence primarily due to their low cost, and the fact that most important staple crops never evolved symbiotic associations with bacteria. In the recent past, advances in our knowledge of symbiosis and nitrogen fixation and the development and application of recombinant DNA technology have created opportunities that could help increase the share of symbiotically-driven nitrogen in global consumption. With the availability of molecular biology tools, rapid improvements in symbiotic characteristics of rhizobial strains became possible. Further, the technology allowed probing the possibility of establishing a symbiotic dialogue between rhizobia and cereals. Because the evolutionary process did not forge a symbiotic relationship with the latter, the potential of molecular manipulations has been tested to incorporate a functional mechanism of nitrogen reduction independent of microbes. In this review, we discuss various strategies applied to improve rhizobial strains for higher nitrogen fixation efficiency, more competitiveness and enhanced fitness under unfavorable environments. The challenges and progress made towards nitrogen self-sufficiency of cereals are also reviewed. An approach to integrate the genetically modified elite rhizobia strains in crop production systems is highlighted.

This paper presents a methodology that uses fishery data collected for the purpose of administering and monitoring harvest quotas in a recreational fishery to give additional insights into effectiveness of various fishing methods, and expected catch rates associated with different licence types. The empirical application is based on the Atlantic salmon (Salmo salar) recreational fishery in Ireland but the statistical analysis is easy to replicate and the models are flexible enough to allow different specifications applicable to other fisheries. The output of the analysis facilitates a better understanding of the factors associated with recreational catches, which in turn provides supplementary information to inform the regulation and management of recreational fisheries.

Bacteria currently included in Rhizobium leguminosarum are too diverse to be considered a single species, so we can refer to this as a species complex (the Rlc). We have found 429 publicly available genome sequences that fall within the Rlc and these show that the Rlc is a distinct entity, well separated from other species in the genus. Its sister taxon is R. anhuiense. We constructed a phylogeny based on concatenated sequences of 120 universal (core) genes, and calculated pairwise average nucleotide identity (ANI) between all genomes. From these analyses, we concluded that the Rlc includes 18 distinct genospecies, plus 7 unique strains that are not placed in these genospecies. Each genospecies is separated by a distinct gap in ANI values, usually at approximately 96% ANI, implying that it is a &lsquo, natural&rsquo, unit. Five of the genospecies include the type strains of named species: R. laguerreae, R. sophorae, R. ruizarguesonis, &ldquo, R. indicum&rdquo, and R. leguminosarum itself. The 16S ribosomal RNA sequence is remarkably diverse within the Rlc, but does not distinguish the genospecies. Partial sequences of housekeeping genes, which have frequently been used to characterize isolate collections, can mostly be assigned unambiguously to a genospecies, but alleles within a genospecies do not always form a clade, so single genes are not a reliable guide to the true phylogeny of the strains. We conclude that access to a large number of genome sequences is a powerful tool for characterizing the diversity of bacteria, and that taxonomic conclusions should be based on all available genome sequences, not just those of type strains.

Gut microbiota have myriad roles in host physiology, development, and immunity. Though confined to the intestinal lumen by the epithelia, microbes influence distal systems via poorly characterized mechanisms. Recent work has considered the role of extracellular vesicles in interspecies communication, but whether they are involved in systemic microbe-host interaction is unclear. Here, we show that distinctive nanoparticles can be isolated from mouse blood within 2.5 h of consuming Lacticaseibacillus rhamnosus JB-1. In contrast to blood nanoparticles from saline-fed mice, they reproduced lipoteichoic acid-mediated immune functions of the original bacteria, including activation of TLR2 and increased IL-10 expression by dendritic cells. Like the fed bacteria, they also reduced IL-8 induced by TNF in an intestinal epithelial cell line. Though enriched for host neuronal proteins, these isolated nanoparticles also contained proteins and viral (phage) DNA of fed bacterial origin. Our data strongly suggest that oral consumption of live bacteria rapidly leads to circulation of their membrane vesicles and phages and demonstrate a nanoparticulate pathway whereby beneficial bacteria and probiotics may systemically affect their hosts.

Memo1deletion in mice causes premature aging and an unbalanced metabolism partially resemblingFgf23andKlotholoss-of-function animals. We report a role for Memo’s redox function in renal FGF23-Klotho signaling using mice with postnatally induced Memo deficiency in the whole body (cKO). Memo cKO mice showed impaired FGF23-driven renal ERK phosphorylation and transcriptional responses. FGF23 actions involved activation of oxidation-sensitive protein phosphotyrosyl phosphatases (PTP) in the kidney. Redox proteomics revealed excessive thiols of Rho-GDP dissociation inhibitor 1 (Rho-GDI1) in Memo cKO, and we detected a functional interaction between Memo’s redox function and oxidation at Rho-GDI1 Cys79. In isolated cellular systems, Rho-GDI1 did not directly affect FGF23-driven cell signaling, but we detected disturbed Rho-GDI1 dependent small Rho-GTPase protein abundance and activity in the kidney of Memo cKO mice. Collectively, this study reveals previously unknown layers in the regulation of renal FGF23 signaling and connects Memo with the network of small Rho-GTPases.

Background Immunosuppressive tumour microenvironments (TME) inhibit the effectiveness of cancer immunotherapies. Sialic acids, which exist as terminal sugars of glyco-conjugates, are highly expressed on cancer cells and are involved in various pathological processes including increased immune evasion, tumour invasiveness and tumour cell metastasis.1 Siglecs (Sialic acid-binding immunoglobulin-type lectins) are expressed on immune cell surfaces and bind sialic acid. Siglec binding to hypersialylated tumour glycans blocks immune cell activation to promote immunosuppression.1 2Intestinal stromal cells (iSCs), precursors to cancer-associated fibroblasts (CAFs), are a key component of the TME and play a vital role in tumour progression by enhancing a tumour-promoting microenvironment. The aim of this study was therefore to investigate if iSC/CAF sialylation contributes to enhanced immunosuppression in the TME. Methods iSCs were isolated from colorectal cancer patient biopsies and cultured ex vivo. Informed consent was obtained from all patients prior to sampling. Tumour-derived iSCs were termed CAFs while control iSCs, isolated from tumour-adjacent non-cancerous tissue, were termed normal-associated fibroblasts (NAFs). NAFs/CAFs were then co-cultured with healthy allogeneic PBMCs and their immunosuppressive properties were assessed by flow cytometry. Results CAFs significantly supressed the proliferation of CD8+ and CD4+ T-cells and induced a more exhausted T-cell phenotype as evidenced by increased expression of the exhaustion markers TIM-3, LAG-3 and PD-1 when compared to co-culture with control NAFs, thereby demonstrating their potent immunosuppressive properties. Strikingly, CAFs also induced significantly higher expression of both Siglec-7 and Siglec-9 receptors on CD8+ T-cells specifically. To elucidate the role of sialylation on CAF-mediated immunosuppression, NAFs/CAFs were treated with the sialyltransferase inhibitor (SI) P-3FAX-Neu5Ac prior to co-culture. Reduction of sialic acid expression on NAFs/CAFs was confirmed by flow cytometry and the SI-treated NAFs/CAFs were then co-cultured with allogeneic T-cells to assess the functional consequences of reduced NAF/CAF sialylation. SI-treated CAFs induced significantly less CD4+TIM-3+ and both CD4+LAG-3+ and CD8+LAG-3+ T-cells compared to their untreated counterparts. Interestingly, SI-treated CAFs also induced significantly less Siglec-7 and -9 receptor-expressing CD8+ T-cells. Conclusions These results demonstrate that non-haematopoietic stromal cells in the tumour-microenvironment can suppress activated T-cells and that this immunosuppressive effect can be significantly reversed through the modulation of sialylation on the stromal cell surface. These results support the hypothesis that stromal cell sialylation plays a role in their immunosuppressive properties. Understanding how sialylation of stromal cells is regulated and functions to enhance immunosuppression in the TME could uncover novel immune checkpoints to reactivate anti-tumour immunity, allowing for tumour cell clearance. Ethics Approval This study was approved by Galway University Hospitals’ Clinical Research Ethics Committee, approval number C.A 2074. Consent N/A References Wang L, Liu Y, Wu L, Sun XL. Sialyltransferase inhibition and recent advances. Biochim Biophys Acta 2016 Jan; 1864(1):143-53. Munkley J, Scott E. Targeting aberrant sialylation to treat cancer. Medicines (Basel) 2019 Oct 13;6(4):102.

The ecology of vector-borne diseases in a region can be attributed to vector-host interactions. In the United States, tick-borne pathogens are the cause of the highest number of reported vector-borne diseases. In the mid-Atlantic region of the eastern United States, tick-borne diseases such as Lyme disease, have increased in incidence, with tick-host-pathogen interactions considered a contributing factor to this increase. Ticks become infected with pathogens after taking a blood meal from a systemically infected host or through a localized infection while co-feeding on a host with other infected ticks. The host not only plays a role in pathogen acquisition by the tick, but can also facilitate dispersal of the tick locally within a region or over greater distances into new geographical ranges outside of their historical distributional range. In this study conducted in southeastern Virginia (USA), we examined the interaction between both resident and migratory bird species and Ixodes ticks, the primary vectors of Borrelia burgdorferi sensu stricto (s.s.) the main causative agent of Lyme disease on the East coast of the United States. Over a two-year period (2012-2014), 1879 passerine birds were surveyed, with 255 Ixodes ticks tested for the presence of Borrelia spp. Eighty passerine birds (4.3 %) representing 17 bird species were parasitized by at least one Ixodes tick, but only three bird species were parasitized by Ixodes ticks that tested positive for B. burgdorferi s.s. Twenty Ixodes ticks (7.8 %) tested positive for B. burgdorferi s.s. with nearly all collected from resident bird species including the Carolina wren (Thryothorus ludovicianus) and brown thrasher (Toxostoma rufum). Given that millions of birds pass through southeastern Virginia during migration, even with the low number of Ixodes ticks parasitizing passerine birds and the low prevalence of B. burgdorferi s.s. found within Ixodes ticks collected, the sheer volume of passerine birds suggests they may play a role in the maintenance and dispersal of B. burgdorferi s.s. in southeastern Virginia.

Mounting stress from multidrug-resistant bacteria has renewed interest in bacteriophage (bacterial virus, phage) therapy as an alternative to antibiotics. While virulent phages quickly infect and kill their host through a lytic lifecycle, temperate phages can stably integrate into the host’s genome until exposed to stressors that can cause a switch to a lytic cycle. Dormant phages can protect their host, known as lysogen, from further phage infections. In vitro this results in rapid host regrowth, which would likely complicate treatment. All case studies and clinical trials to date use virulent phages. However, temperate phages are far easier to find and more abundant in (and on) humans, playing a vital role in vivo. To make use of these abundant entities, we propose a synergy using stressors such as antibiotics, which are known to awaken dormant phages. Here we demonstrate that co-administration of the temperate phage HK97 with sub-inhibitory concentrations of a fluoroquinolone antibiotic, ciprofloxacin, results in bacterial eradication (≥8 log reduction) in vitro. This synergy is up to 4.5 x105 fold greater than the multiplicative effects of the phage and antibiotic alone. Furthermore, there is no large increase in final phage titre, latency period or burst size – as such, this synergy is distinct from traditional phage-antibiotic-synergy described with virulent phages. We demonstrate the presence of the antibiotic prevents lysogen colony formation over time, the biggest hurdle in the current use of temperate phages in therapy. As the interaction between temperate phages and stressors such as ciprofloxacin are known to be widespread, the findings from this work may allow for the use of these phages in combination with antibiotics for the treatment of antimicrobial-resistant infections.

Endocrine disrupting chemicals (EDCs) are exogenous substances that interfere with the endocrine system and cause adverse effects. We aimed to classify the effects of 24 known EDCs, prevalent in certain occupations, according to four modes of action (estrogenic, antiestrogenic, androgenic, and/or antiandrogenic). A literature search, stratified into four types of literature was conducted (namely: national and international agency reports; review articles; primary studies; ToxCastTM). The state of the evidence of each EDC on sex hormone function was summarized and reviewed by an expert panel. For each mode of action, the experts evaluated the likelihood of endocrine disruption in five categories: “No”, “Unlikely”, “Possibly”, “Probably”, and “Yes”. Seven agents were categorized as “Yes,” or having strong evidence for their effects on sex hormone function (antiandrogenic: lead, arsenic, butylbenzyl phthalate, dibutyl phthalate, dicyclohexyl phthalate; estrogenic: nonylphenol, bisphenol A). Nine agents were categorized as “Probable,” or having probable evidence (antiandrogenic: bis(2-ethylhexyl)phthalate, nonylphenol, toluene, bisphenol A, diisononyl phthalate; androgenic: cadmium; estrogenic: copper, cadmium and; anti-estrogenic: lead). Two agents (arsenic, polychlorinated biphenyls) had opposing conclusions supporting both “probably” estrogenic and antiestrogenic effects. This synthesis will allow researchers to evaluate the health effects of selected EDCs with an added level of precision related to the mode of action.

Coordination of a number of molecular mechanisms including transcription, alternative splicing, and class switch recombination are required to facilitate development, activation, and survival of B cells. Disruption of these pathways can result in malignant transformation. Recently, next-generation sequencing has identified a number of novel mutations in mantle cell lymphoma (MCL) patients including mutations in the ubiquitin E3 ligase UBR5. Approximately 18% of MCL patients were found to have mutations in UBR5, with the majority of mutations within the HECT domain of the protein that can accept and transfer ubiquitin molecules to the substrate. Determining if UBR5 controls the maturation of B cells is important to fully understand malignant transformation to MCL. To elucidate the role of UBR5 in B-cell maturation and activation, we generated a conditional mutant disrupting UBR5′s C-terminal HECT domain. Loss of the UBR5 HECT domain leads to a block in maturation of B cells in the spleen and upregulation of proteins associated with messenger RNA splicing via the spliceosome. Our studies reveal a novel role of UBR5 in B-cell maturation by stabilization of spliceosome components during B-cell development and suggests UBR5 mutations play a role in MCL transformation.

Introduction International travellers contribute to the rapid spread of Zika virus (ZIKV) and its sentinel identification globally. We describe ZIKV infections among international travellers seen at GeoSentinel sites with a focus on ZIKV acquired in the Americas and the Caribbean, describe countries of exposure and traveller characteristics, and assess ZIKV diagnostic testing by site. Methods Records with an international travel-related diagnosis of confirmed or probable ZIKV from January 2012 through December 2019 reported to GeoSentinel with a recorded illness onset date were included to show reported cases over time. Records from March 2016 through December 2019 with an exposure region of the Americas or the Caribbean were included in the descriptive analysis. A survey was conducted to assess the availability, accessibility and utilization of ZIKV diagnostic tests at GeoSentinel sites. Results GeoSentinel sites reported 525 ZIKV cases from 2012 through 2019. Between 2012 and 2014, eight cases were reported, and all were acquired in Asia or Oceania. After 2014, most cases were acquired in the Americas or the Caribbean, a large decline in ZIKV cases occurred in 2018–19. Between March 2016 and December 2019, 423 patients acquired ZIKV in the Americas or the Caribbean, peak reporting to these regions occurred in 2016 [330 cases (78%)]. The median age was 36 years (range: 3–92); 63% were female. The most frequent region of exposure was the Caribbean (60%). Thirteen travellers were pregnant during or after travel; one had a sexually acquired ZIKV infection. There was one case of fetal anomaly and two travellers with Guillain-Barré syndrome. GeoSentinel sites reported various challenges to diagnose ZIKV effectively. Conclusion ZIKV should remain a consideration for travellers returning from areas with risk of ZIKV transmission. Travellers should discuss their travel plans with their healthcare providers to ensure ZIKV prevention measures are taken.

Women and men can differ in their propensity to take risks and develop impulse control and addiction disorders. Sexual dimorphisms in behavioral control by the mesolimbic dopaminergic reward system may underlie these phenomena, given its sensitivity to gonadal hormones. However, this is hard to test experimentally using humans. Using the rat gambling task (rGT), we investigated what impact acute inhibition of accumbal dopamine had on decision-making and impulsivity in animals of both sexes. We expressed an inhibitory designer receptor exclusively activated by designer drugs (hM4D[Gi]) in the accumbal dopaminergic efferents of female and male transgenic (Tg) rats, engineered to selectively express cre recombinase in neurons synthesizing tyrosine hydroxylase. We then trained the rats to perform the rGT and assessed the effect of an acute clozapine-n-oxide (0-3 mg/kg) challenge. Chemogenetic inhibition of dopaminergic projections to the accumbens did not affect choice in females, perhaps due to low levels of risky choice in Tg+ animals at baseline, but induced a switch from risky to optimal decision-making in males performing the cued rGT. This manipulation also decreased motor impulsivity but only in females. These data support the hypothesis that cue-driven risky choice is mediated, at least in males, by activity of accumbal dopaminergic neurons. However, motor impulsivity is more sensitive to inhibition of accumbal dopamine neurons in female rats. These data may help explain differences in the manifestation of addictions across gender and reinforce the importance of examining both sexes when seeking to attribute control of behavior to specific monoaminergic pathways. (PsycInfo Database Record (c) 2020 APA, all rights reserved).

Hematopoiesis is responsible for numerous functions, ranging from oxygen transportation to host defense, to injury repair. This process of hematopoiesis is maintained throughout life by hematopoietic stem cells and requires a controlled balance between self-renewal, differentiation, and quiescence. Disrupting this balance can result in hematopoietic malignancies, including anemia, immune deficiency, leukemia, and lymphoma. Recent work has shown that FBOX E3 ligases, a substrate recognition component of the ubiquitin proteasome system (UPS), have an integral role in maintaining this balance. In this review, we detail how FBOX proteins target specific proteins for degradation to regulate hematopoiesis through cell processes, such as cell cycle, development, and apoptosis.

Phages vB_RleM_RL38JI and vB_RleM_RL2RES are known to mediate generalized transduction in Rhizobium leguminosarum. The RL38JI genome consists of 158,577 nucleotides and 270 predicted genes, whereas RL2RES has a 156,878-bp genome with 262 predicted genes. The two genomes are similar, with 82.88% nucleotide identity to each other.

Notch is a cell–cell signaling pathway that is involved in a host of activities including development, oncogenesis, skeletal homeostasis, and much more. More specifically, recent research has demonstrated the importance of Notch signaling in osteogenic differentiation, bone healing, and in the development of the skeleton. The craniofacial skeleton is complex and understanding its development has remained an important focus in biology. In this review we briefly summarize what recent research has revealed about Notch signaling and the current understanding of how the skeleton, skull, and face develop. We then discuss the crucial role that Notch plays in both craniofacial development and the skeletal system, and what importance it may play in the future.

Summary Bacteriophages encoding anti-CRISPR proteins (Acrs) must cooperate to overcome phage resistance mediated by the bacterial immune system CRISPR-Cas, where the first phage blocks CRISPR-Cas immunity in order to allow a second Acr phage to successfully replicate. However, in nature, bacteria are frequently not pre-immunized, and phage populations are often not clonal, exhibiting variations in Acr presence and strength. We explored how interactions between Acr phages and initially sensitive bacteria evolve, both in the presence and absence of competing phages lacking Acrs. We find that Acr phages benefit “Acr-negative” phages by limiting the evolution of CRISPR-based resistance and helping Acr-negative phages to replicate on resistant host sub-populations. These benefits depend on the strength of CRISPR-Cas inhibitors and result in strong Acrs providing smaller fitness advantages than weaker ones when Acr phages compete with Acr-negative phages. These results indicate that different Acr types shape the evolutionary dynamics and social interactions of phage populations in natural communities., Graphical Abstract, Highlights • Acr-positive phages limit evolution of CRISPR resistance during clonal and mixed infections • Acr-positive phages provide benefits to Acr-negative phages present in the community • Strong Acr help Acr-negative phages to amplify on immunosuppressed CRISPR-resistant cells • Weaker Acr provides larger advantages during competition with Acr-negative phages, Chevallereau et al. find that phages encoding anti-CRISPR proteins (Acrs) protect the larger phage community, including Acr-negative phages, from bacterial CRISPR-Cas immunity by immunosuppressing resistant cells and limiting immune adaptation. Because they do not allow for such exploitation, low-efficiency Acrs provide a greater advantage during competition with Acr-negative phages.

Multiple treatments for dorsal wrist ganglia (DWG) exist but have high recurrence rates. We investigated whether aspiration followed by injection of Tisseel is more effective at reducing the DWG recurrence rate than aspiration alone.Adults with untreated DWG were randomly assigned to aspiration alone (A) or aspiration followed by Tisseel injection (AT). Patients were measured at baseline, 1, 6, and 12 months. Primary outcome was the rate of recurrence. Secondary outcomes included recurrent ganglion size and maximum pain experienced from the ganglion. Continuous data were compared with StudentSeventy-nine patients were randomized to the A (n = 39) or AT (n = 40) group. Twenty-five patients were lost to follow-up. There were no differences in recurrence risk between groups at 1, 6, or 12 months (Aspiration and Tisseel injection does not decrease DWG recurrence versus aspiration alone. Both interventions produced a significant decrease in pain. Although aspiration alone carries a high recurrence risk, it may provide sufficient symptomatic relief for patients with DWG.Malgré les nombreux traitements des ganglions de la face dorsale du poignet (GFDP), les taux de récurrence sont élevés. Les auteurs ont évalué si la ponction suivie d’une injection de Tisseel réduisait le taux de récurrence de GFDP avec plus d’efficacité que la ponction seule.Les adultes présentant un GFDP non traité ont été répartis au hasard entre la ponction seule (P) et la ponction suivie par une injection de Tisseel (PT). Les chercheurs les ont mesurés au départ, puis au bout de un, six et 12 mois. Le taux de récurrence représentait le résultat clinique primaire, et la taille des ganglions récurrents et la douleur maximale provoquée par le ganglion, les résultats cliniques secondaires. Les chercheurs ont comparé les données continues avec les tests de Student et les tests Mann-Whitney et les données catégorielles avec les tests du chi carré et les tests exacts de Fisher. Ils ont déclaré l’effet du traitement sous forme de risque relatif (RR) ou de différences moyennes (DM), selon des intervalles de confiance à 95 %.Au total, les chercheurs ont réparti 79 patients au hasard entre le groupe P (n = 39) ou PT (n = 40). Ils ont perdu 25 patients au suivi. Ils n’ont constaté aucune différence quant au risque de récurrence entre les groupes au bout de un, six ou 12 mois (La ponction et l’injection de Tisseel ne réduisent pas la récurrence de GFDP par rapport à la ponction seule. Les deux interventions entraînaient une diminution significative de la douleur. Même si la ponction seule s’associe à un fort risque de récurrence, elle peut apporter un soulagement symptomatique suffisant pour les patients ayant un GFDP.

The development of new treatments for castrate resistant prostate cancer (CRPC) must address such challenges as intrinsic tumor heterogeneity and phenotypic plasticity. Combined PTEN/TP53 alterations represent a major genotype of CRPC (25–30%) and are associated with poor outcomes. Using tumor-derived, castration-resistant Pten/Tp53 null luminal prostate cells for comprehensive, high-throughput, mechanism-based screening, we identified several vulnerabilities among >1900 compounds, including inhibitors of: PI3K/AKT/mTOR, the proteasome, the cell cycle, heat shock proteins, DNA repair, NFκB, MAPK, and epigenetic modifiers. HSP90 inhibitors were one of the most active compound classes in the screen and have clinical potential for use in drug combinations to enhance efficacy and delay the development of resistance. To inform future design of rational drug combinations, we tested ganetespib, a potent second-generation HSP90 inhibitor, as a single agent in multiple CRPC genotypes and phenotypes. Ganetespib decreased growth of endogenous Pten/Tp53 null tumors, confirming therapeutic activity in situ. Fifteen human CRPC LuCaP PDX-derived organoid models were assayed for responses to 110 drugs, and HSP90 inhibitors (ganetespib and onalespib) were among the select group of drugs (75% of models) at high potency (IC50