Bone marrow-derived cells are recruited into tumor vasculature in response to angiogenic signals, and some of the cells within the newly forming tumor vessels are hematopoietic stem cells (HSCs) in origin. Previous studies suggest that bone marrow-derived pericytes are associated with newly formed vessels in tumors. In this study, we used an orthotopic rat glioma model (RT-2/RAG) to examine the contribution of long-term hematopoietic stem cell (LT-HSC)-derived pericytic cells to brain tumor angiogenesis. Mice (RAG-2/KO5.2) were lethally irradiated, and their hematopoietic cells were repopulated by transplantation of double fluorescence-activated cell-sorted LT-HSCs that express green fluorescent protein (GFP+). RT-2/RAG cells were then injected into the striatum of the chimeric mice 6 weeks post-transplantation. The animals were sacrificed 9 days after tumor implantation, and the incorporation and lineage-specific marker expression profile of the GFP+ cells within the growing tumor and tumor periphery were analyzed. LT-HSC-derived GFP+ cells were noted to incorporate onto the surface of tumor vessels within the perivascular space. LT-HSC-derived GFP+ cells express the pericyte progenitor marker, platelet-derived growth factor receptor-beta (PDGFR beta), as well as mature perictyte markers such as nerve/glial antigen 2 proteoglycan (NG2), alpha-smooth muscle actin (alpha SMA), and desmin. These LT-HSC-derived cells may represent a population of progenitor or committed pericytes within the neovascular tree and may play a role in shaping the angio-architecture in the vascular niche of brain tumors.

Reactivities of the monoclonal antibodies (mAbs) of the 9th Human Leukocyte Differentiation Antigen Workshop, in order to define specific antigenic expression of the primary cutaneous B-cell lymphomas (PC-BCL), were analyzed by immunohistology on human tonsil and on PC-BCL, such as follicular centre B-cell lymphomas (FCL), marginal zone lymphomas (MZL) and diffuse large B-cells lymphomas leg-type (DLBL-LT). We identified some subgroups of mAbs that were exclusively or preferentially positive in one lymphoma cell type: the PC-FCL subgroup of mAbs includes PD1/CD279, GCET-1, hFCRL1/CD307a, FCRL2/CD307b, CXCR5/CD185, BBTLA/CD272, BLIMP-1, hCD38. No specific subgroup of mAbs was found to label PC-DLBCL. This study may be useful to better define specific antigen profile of different PC-BCL entities leading to a correct diagnosis.

BACKGROUND

Neuroinflammation and protein accumulation are characteristic hallmarks of both normal aging and age-related neurodegenerative diseases. However, the relationship between these factors in neurodegenerative processes is poorly understood. We have previously shown that proteasome inhibition produced higher neurodegeneration in aged than in young rats, suggesting that other additional age-related events could be involved in neurodegeneration. We evaluated the role of lipopolysaccharide (LPS)-induced neuroinflammation as a potential synergic risk factor for hippocampal neurodegeneration induced by proteasome inhibition.

METHODS

Young male Wistar rats were injected with 1 μL of saline or LPS (5 mg/mL) into the hippocampus to evaluate the effect of LPS-induced neuroinflammation on protein homeostasis. The synergic effect of LPS and proteasome inhibition was analyzed in young rats that first received 1 μL of LPS and 24 h later 1 μL (5 mg/mL) of the proteasome inhibitor lactacystin. Animals were sacrificed at different times post-injection and hippocampi isolated and processed for gene expression analysis by real-time polymerase chain reaction; protein expression analysis by western blots; proteasome activity by fluorescence spectroscopy; immunofluorescence analysis by confocal microscopy; and degeneration assay by Fluoro-Jade B staining.

RESULTS

LPS injection produced the accumulation of ubiquitinated proteins in hippocampal neurons, increased expression of the E2 ubiquitin-conjugating enzyme U<em>B</em>2L6, decreased proteasome activity and increased immunoproteasome content. However, LPS injection was not sufficient to produce neurodegeneration. The combination of neuroinflammation and proteasome inhibition leads to higher neuronal accumulation of ubiquitinated proteins, predominant expression of pro-apoptotic markers and increased neurodegeneration, when compared with LPS or lactacystin (LT) injection alone.

CONCLUSIONS

Our results identify neuroinflammation as a risk factor that increases susceptibility to neurodegeneration induced by proteasome inhibition. These results highlight the modulation of neuroinflammation as a mechanism for neuronal protection that could be relevant in situations where both factors are present, such as aging and neurodegenerative diseases.

<A<em>b</em>stractText>Studies in mice have shown that the intestinal micro<em>b</em>iota can contri<em>b</em>ute to o<em>b</em>esity via the anorexigenic gut hormone glucagon-like peptide 1 (GLP1) and <em>b</em>ile acids, which affect lipid meta<em>b</em>olism. We performed a randomized, place<em>b</em>o-controlled, pilot study of the effects of fecal micro<em>b</em>iota transplantation (FMT) in o<em>b</em>ese, meta<em>b</em>olically uncompromised patients.</A<em>b</em>stractText><p><div>(<em>b</em>)METHODS</<em>b</em>)</div>We performed a dou<em>b</em>le-<em>b</em>lind study of 22 o<em>b</em>ese patients (<em>b</em>ody mass index [BMI] ≥5 kg/m²) without a diagnosis of dia<em>b</em>etes, nonalcoholic steatohepatitis, or meta<em>b</em>olic syndrome. Participants were assigned randomly (1:1) to groups that received FMT <em>b</em>y capsules (induction dose of 30 capsules at week 4 and maintenance dose of 12 capsules at week 8) or place<em>b</em>o capsules. FMT capsules were derived from a single lean donor (BMI, 17.5 kg/m²). Patients were followed up through week 26; the primary outcome was safety. Stool and serum samples were collected from patients at <em>b</em>aseline and at weeks 1, 4, 6, 8, and 12 after administration of the first dose of FMT or place<em>b</em>o and analyzed <em>b</em>y 16S RNA gene sequencing. Stool and serum samples were analyzed for meta<em>b</em>olomics <em>b</em>y liquid chromatography-mass spectrometry. Additional outcomes were the change in area under the curve for GLP1 at week 12.</p><A<em>b</em>stractText>We o<em>b</em>served no significant differences in adverse events <em>b</em>etween patients who received FMT vs place<em>b</em>o. There was no increase in the area under the curve of GLP1 in either group. Patients who received FMT had sustained shifts in micro<em>b</em>iomes associated with o<em>b</em>esity toward those of the donor (P &<em>lt</em>; .001). Patients who received FMT had a sustained decrease in stool levels of taurocholic acid (P &<em>lt</em>; .05) compared with <em>b</em>aseline; <em>b</em>ile acid profiles <em>b</em>egan to resem<em>b</em>le those of the donor more closely. We did not o<em>b</em>serve significant changes in mean BMI at week 12 in either group.</A<em>b</em>stractText><A<em>b</em>stractText>In a place<em>b</em>o-controlled pilot study, we found that FMT capsules (derived from a lean donor) were safe <em>b</em>ut did not reduce BMI in o<em>b</em>ese meta<em>b</em>olically uncompromised patients. The FMT capsules were well tolerated and led to sustained changes in the intestinal micro<em>b</em>iome and <em>b</em>ile acid profiles that were similar to those of the lean donor. ClinicalTrials.gov num<em>b</em>er: NCT02741518.</A<em>b</em>stractText>

(<em>b</em>)Background:</<em>b</em>) Patients with acutely decompensated cirrhosis (AD) may or may not develop acute-on-chronic liver failure (ACLF). ACLF is characterized <em>b</em>y high-grade systemic inflammation, organ failures (OF) and high short-term mortality. A<em>lt</em>hough patients with AD cirrhosis exhi<em>b</em>it distinct clinical phenotypes at <em>b</em>aseline, they have low short-term mortality, unless ACLF develops during follow-up. Because little is known a<em>b</em>out the association of profile of systemic inflammation with clinical phenotypes of patients with AD cirrhosis, we aimed to investigate a <em>b</em>attery of markers of systemic inflammation in these patients. (<em>b</em>)Methods:</<em>b</em>) Upon hospital admission <em>b</em>aseline plasma levels of 15 markers (cytokines, chemokines, and oxidized al<em>b</em>umin) were measured in 40 hea<em>lt</em>hy controls, 39 compensated cirrhosis, 342 AD cirrhosis, and 161 ACLF. According to EASL-CLIF criteria, AD cirrhosis was divided into three distinct clinical phenotypes (AD-1: Creatinine&<em>lt</em>;1.5, no HE, no OF; AD-2: creatinine 1.5-2, and or HE grade I/II, no OF; AD-3: Creatinine&<em>lt</em>;1.5, no HE, non-renal OF). (<em>b</em>)Resu<em>lt</em>s:</<em>b</em>) Most markers were slightly a<em>b</em>normal in compensated cirrhosis, <em>b</em>ut markedly increased in AD. Patients with ACLF exhi<em>b</em>ited the largest num<em>b</em>er of a<em>b</em>normal markers, indicating "full-<em>b</em>lown" systemic inflammation (all markers). AD-patients exhi<em>b</em>ited distinct systemic inflammation profiles across three different clinical phenotypes. In each phenotype, activation of systemic inflammation was only partial (30% of the markers). Mortality related to each clinical AD-phenotype was significantly lower than mortality associated with ACLF (<i>p</i> &<em>lt</em>; 0.0001 <em>b</em>y gray test). Among AD-patients <em>b</em>aseline systemic inflammation (especially IL-8, IL-6, IL-1ra, HNA2 independently associated) was more intense in those who had poor 28-day outcomes (ACLF, death) than those who did not experience these outcomes. (<em>b</em>)Conclusions:</<em>b</em>) A<em>lt</em>hough AD-patients exhi<em>b</em>it distinct profiles of systemic inflammation depending on their clinical phenotypes, all these patients have only partial activation of systemic inflammation. However, those with the most extended <em>b</em>aseline systemic inflammation had the highest the risk of ACLF development and death.

<p><div>(<em>b</em>)PURPOSE</<em>b</em>)</div>To assess talazopari<em>b</em> activity in germline <i>BRCA1/2</i> mutation carriers with advanced <em>b</em>reast cancer.</p><p><div>(<em>b</em>)PATIENTS AND METHODS</<em>b</em>)</div>ABRAZO (NCT02034916) was a two-cohort, two-stage, phase II study of talazopari<em>b</em> (1 mg/day) in germline <i>BRCA</i> mutation carriers with a response to prior platinum with no progression on or within 8 weeks of the last platinum dose (cohort 1) or ≥3 platinum-free cytotoxic regimens (cohort 2) for advanced <em>b</em>reast cancer. Primary endpoint was confirmed o<em>b</em>jective response rate (ORR) <em>b</em>y independent radiological assessment.</p><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>We enrolled 84 patients (cohort 1, <i>n</i> = 49; cohort 2, <i>n</i> = 35) from May 2014 to Fe<em>b</em>ruary 2016. Median age was 50 (range, 31-75) years. Triple-negative <em>b</em>reast cancer (TNBC) incidence was 59% (cohort 1) and 17% (cohort 2). Median num<em>b</em>er of prior cytotoxic regimens for advanced <em>b</em>reast cancer was two and four, respectively. Confirmed ORR was 21% [95% confidence interval (CI), 10-35; cohort 1] and 37% [95% CI, 22-55; cohort 2]. Median duration of response was 5.8 and 3.8 months, respectively. Confirmed ORR was 23% (<i>BRCA1</i>), 33% (<i>BRCA2</i>), 26% (TNBC), and 29% (hormone receptor-positive). The most common all-grade adverse events (AE) included anemia (52%), fatigue (45%), and nausea (42%). Talazopari<em>b</em>-related AEs led to drug discontinuation in 3 (4%) patients. In an exploratory analysis, longer platinum-free interval was associated with higher response rate in cohort 1 (0% ORR with interval &<em>lt</em>;8 weeks; 47% ORR with interval >6 months).</p><p><div>(<em>b</em>)CONCLUSIONS</<em>b</em>)</div>Talazopari<em>b</em> exhi<em>b</em>ited promising antitumor activity in patients with advanced <em>b</em>reast cancer and germline <i>BRCA</i> mutation.</p>

Marbling is an important criterion for beef quality grading in many countries. The purpose of the current study was to utilize the natural genetic variation to identify major metabolic indicators of marbling in cattle differing in genotypes. Rectus abdominis (RA, oxidative), semitendinosus (glycolytic), and longissimus thoracis (LT, oxido-glycolytic) muscles were taken from steers of different genotypes that expressed high [Angus, n = 16; and crossbred (Angus x Japanese Black), n = 10] or low (Limousin, n = 12) levels of marbling in their meat. Muscles from Angus and crossbred steers were characterized, as expected, by a greater triacylglycerol (TAG) content (P < 0.001) and also by greater protein contents of fatty acid-binding protein specific for heart and muscles (H-FABP; P < 0.001 for RA and P < 0.05 for LT muscle) or for adipocytes (A-FABP; P < 0.001 for RA and LT muscles). Moreover, oxidative enzyme activities (beta-hydroxyacyl-CoA dehydrogenase, citrate synthase, isocitrate dehydrogenase, cytochrome-c oxidase) were greater (P < 0.01 to 0.001) in the 3 muscles studied, whereas glycolytic enzyme activities (phosphofructokinase and lactate dehydrogenase) were lower (P < 0.001) in RA muscle in Angus and crossbred steers compared with Limousin steers. Significant correlations were observed between TAG content and H- and A-FABP protein contents, and oxidative (r>> or = +0.55, P < 0.001) or glycolytic enzyme activities (r>> or = -0.47, P < 0.001), when the 3 genotypes and muscles studied were considered as a whole. In addition, A-FABP protein content and some oxidative enzyme activities were significantly correlated with TAG content independently of the genotype and muscle effects. In conclusion, A-FABP protein content, as well as oxidative enzyme activities, may be used as indicators of the ability of steers from extreme genotypes to deposit intramuscular fat.

The synthesis of selected antigens in plants and their oral delivery has great potential for reducing the costs of vaccine production and administration. The application of this technology requires antigen concentrations in final plant material to be uniform to ensure consistent dosing. In addition, antigen levels should be such as to allow the volume of each dose, containing a set amount of antigen, to be practical for oral delivery. Here, we demonstrate that the Lt-B protein of enterotoxigenic E. coli is evenly distributed in defatted corn germ prepared from transgenic grain. Furthermore, the choice of sub-cellular location for Lt-B affects accumulation of the protein in excess of four orders of magnitude.

<A<em>b</em>stractText>The aim of this study was to evaluate the low-density lipoprotein cholesterol lowering efficacy and safety of a <em>b</em>empedoic acid 180 mg and ezetimi<em>b</em>e 10 mg fixed-dose com<em>b</em>ination in patients with hypercholesterolemia and a high risk of cardiovascular disease receiving maximally tolerated statin therapy.</A<em>b</em>stractText><A<em>b</em>stractText>This phase 3, dou<em>b</em>le-<em>b</em>lind clinical trial enrolled adu<em>lt</em> patients at high risk of cardiovascular disease due to atherosclerotic cardiovascular disease, heterozygous familial hypercholesterolemia, or mu<em>lt</em>iple cardiovascular disease risk factors. Patients were randomly assigned (2:2:2:1) to treatment with the fixed-dose com<em>b</em>ination, <em>b</em>empedoic acid 180 mg, ezetimi<em>b</em>e 10 mg or place<em>b</em>o added to sta<em>b</em>le <em>b</em>ackground statin therapy for 12 weeks. The primary efficacy endpoint was the percentage change from <em>b</em>aseline to week 12 in low-density lipoprotein cholesterol.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>Among the 301 patients included in the primary analysis, the mean <em>b</em>aseline low-density lipoprotein cholesterol level was 3.87 mmol/L (149.8 mg/dL). At week 12, the fixed-dose com<em>b</em>ination lowered low-density lipoprotein cholesterol (-36.2%) significantly more than place<em>b</em>o (1.8% (place<em>b</em>o-corrected difference -38.0%); <i>P</i> &<em>lt</em>; 0.001), ezetimi<em>b</em>e alone (-23.2%; <i>P</i> &<em>lt</em>; 0.001) or <em>b</em>empedoic acid alone (-17.2%; <i>P</i> &<em>lt</em>; 0.001). The fixed-dose com<em>b</em>ination lowered low-density lipoprotein cholesterol levels similarly across su<em>b</em>groups, including patients receiving high-intensity, other-intensity or no statin therapy. Improvements with the fixed-dose com<em>b</em>ination were also o<em>b</em>served in secondary efficacy endpoints, including high-sensitivity C-reactive protein. In this trial, fixed-dose com<em>b</em>ination treatment had a generally similar safety profile compared with <em>b</em>empedoic acid, ezetimi<em>b</em>e or place<em>b</em>o.</p><A<em>b</em>stractText>The <em>b</em>empedoic acid and ezetimi<em>b</em>e fixed-dose com<em>b</em>ination significantly lowered low-density lipoprotein cholesterol versus place<em>b</em>o or other oral monotherapies and had a favoura<em>b</em>le safety profile when added to maximally tolerated statin therapy in patients with hypercholesterolemia and high cardiovascular disease risk.</A<em>b</em>stractText><A<em>b</em>stractText>ClinicalTrials.gov identifier: NCT03337308.</A<em>b</em>stractText>

The B6.SJL-Ptprc(d)Pep3(b)/BoyJ (B6.SJL) congenic mouse strain, a valuable and widely used tool in murine bone marrow transplantation studies, has long been considered equivalent to the parental C57B/L6 (B6) strain with the exception of a small congenic interval on chromosome 1 harboring an alternative CD45/Ly-5 alloantigen (Ly-5.1). In this study we compared functional properties of stem and stromal cells between the strains, and delineated the boundary of the B6.SJL congenic interval. We identified a 25% reduction in homing efficiency, 3.8-fold reduction in transplantable long-term hematopoietic stem cells (LT-HSCs), a 5-fold reduction in LT-HSCs capable of 24-hour homing, and a cell-intrinsic engraftment defect of 30% to 50% in B6.SJL-derived bone marrow cells relative to B6-derived cells. These functional differences were independent of stem cell number, cycling, or apoptosis. Genotypic analysis revealed a 42.1-mbp congenic interval in B6.SJL including 306 genes, and at least 124 genetic polymorphisms. Moreover, expression profiling revealed 288 genes differentially expressed between nonhematopoietic stromal cells of the 2 strains. These results indicate that polymorphisms between the B6 and SJL genotype within the B6.SJL congenic interval influence HSC engraftment and result in transcriptional variation within bone marrow stroma.

(<em>b</em>)Rationale:</<em>b</em>) Given the rapid spread of COVID-19, an updated risk-stratify prognostic tool could help clinicians identify the high-risk patients with worse prognoses. We aimed to develop a non-invasive and easy-to-use prognostic signature <em>b</em>y chest CT to individually predict poor outcome (death, need for mechanical ventilation, or intensive care unit admission) in patients with COVID-19. (<em>b</em>)Methods:</<em>b</em>) From Novem<em>b</em>er 29, 2019 to Fe<em>b</em>ruary 19, 2020, a total of 492 patients with COVID-19 from four centers were retrospectively collected. Since different durations from symptom onsets to the first CT scanning might affect the prognostic model, we designated the 492 patients into two groups: 1) the early-phase group: CT scans were performed within one week after symptom onset (0-6 days, n = 317); and 2) the late-phase group: CT scans were performed one week later after symptom onset (≥7 days, n = 175). In each group, we divided patients into the primary cohort (n = 212 in the early-phase group, n = 139 in the late-phase group) and the external independent validation cohort (n = 105 in the early-phase group, n = 36 in the late-phase group) according to the centers. We <em>b</em>ui<em>lt</em> two separate radiomics models in the two patient groups. Firstly, we proposed an automatic segmentation method to extract lung volume for radiomics feature extraction. Secondly, we applied several image preprocessing procedures to increase the reproduci<em>b</em>ility of the radiomics features: 1) applied a low-pass Gaussian fi<em>lt</em>er <em>b</em>efore voxel resampling to prevent aliasing; 2) conducted ComBat to harmonize radiomics features per scanner; 3) tested the sta<em>b</em>ility of the features in the radiomics signature <em>b</em>y several image transformations, such as rotating, translating, and growing/shrinking. Thirdly, we used least a<em>b</em>solute shrinkage and selection operator (LASSO) to <em>b</em>uild the radiomics signature (RadScore). Afterward, we conducted a Fine-Gray competing risk regression to <em>b</em>uild the clinical model and the clinic-radiomics signature (CrrScore). Finally, performances of the three prognostic signatures (clinical model, RadScore, and CrrScore) were estimated from the two aspects: 1) cumulative poor outcome pro<em>b</em>a<em>b</em>ility prediction; 2) 28-day poor outcome prediction. We also did stratified analyses to explore the potential association <em>b</em>etween the CrrScore and the poor outcomes regarding different age, type, and comor<em>b</em>idity su<em>b</em>groups. (<em>b</em>)Resu<em>lt</em>s:</<em>b</em>) In the early-phase group, the CrrScore showed the <em>b</em>est performance in estimating poor outcome (C-index = 0.850), and predicting the pro<em>b</em>a<em>b</em>ility of 28-day poor outcome (AUC = 0.862). In the late-phase group, the RadScore alone achieved similar performance to the CrrScore in predicting poor outcome (C-index = 0.885), and 28-day poor outcome pro<em>b</em>a<em>b</em>ility (AUC = 0.976). Moreover, the RadScore in <em>b</em>oth groups successfully stratified patients with COVID-19 into low- or high-RadScore groups with significantly different survival time in the training and validation cohorts (all <i>P</i> &<em>lt</em>; 0.05). The CrrScore in <em>b</em>oth groups can also significantly stratify patients with different prognoses regarding different age, type, and comor<em>b</em>idities su<em>b</em>groups in the com<em>b</em>ined cohorts (all <i>P</i> &<em>lt</em>; 0.05). (<em>b</em>)Conclusions:</<em>b</em>) This research proposed a non-invasive and quantitative prognostic tool for predicting poor outcome in patients with COVID-19 <em>b</em>ased on CT imaging. Taking the insufficient medical recourse into account, our study might suggest that the chest CT radiomics signature of COVID-19 is more effective and ideal to predict poor outcome in the late-phase COVID-19 patients. For the early-phase patients, integrating radiomics signature with clinical risk factors can achieve a more accurate prediction of individual poor prognostic outcome, which ena<em>b</em>les appropriate management and surveillance of COVID-19.

Keywords: COVID-19; Computed tomography; Poor outcome; Prognosis; Radiomics.

BACKGROUND

Recently it has been demonstrated that after selenium (Se) supplementation in autoimmune thyroiditis (AIT) patients, there was a significant decrease of thyroid peroxidase (TPO) autoantibody (TPOAb) levels. The aim of our study was to evaluate the immunological benefit of Se administration in unselected AIT patients and thus address the question whether Se administration should generally be recommended for AIT patients.

METHODS

Thirty-six consecutive AIT patients (aged 19-85 years) were included in the present study. In addition to their levothyroxine (LT(4)) treatment, 18 patients received 200 microg (2.53 micromol) sodium selenite per day orally for the time span of 3 months, whereas 18 patients received placebo. All patients had measurement of thyroid hormones, thyrotropin (TSH), autoantibodies (thyroglobulin antibodies [TgAb] and TPOAb), Se levels, and intracellular cytokine detection in CD4(+) and CD8(+) T cells of peripheral blood mononuclear cells (PBMC) by flow cytometry before and after Se or placebo administration.

RESULTS

No significant difference in the TPOAb levels was found after Se administration (524 +/- 452 vs. 505 +/- 464 IU/mL; p>> 0.05). Furthermore, we found no significant differences in the CD4(+) or CD8(+) cytokine pattern (IFN-gamma, IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, TNF-alpha, TNF-beta) in patients before and after Se administration, in patients before and after placebo administration and between Se group and placebo group before and after Se vs. placebo administration.

CONCLUSIONS

We demonstrate that Se administration in our AIT patient's cohort does not induce significant immunological changes, either in terms of cytokine production patterns of peripheral T lymphocytes or of TPOAb levels. Our data suggest that AIT patients with moderate disease activity (in terms of TPOAb and cytokine production patterns) may not (equally) benefit as patients with high disease activity.

<A<em>b</em>stractText>The range of <em>b</em>enefits associated with regular physical activity participation is irrefuta<em>b</em>le. Despite the well-known <em>b</em>enefits, physical inactivity remains one of the major contri<em>b</em>uting factors to ill-hea<em>lt</em>h throughout industrialized countries. Traditional lifestyle interventions such as group education or telephone counseling are effective at increasing physical activity participation; however, physical activity levels tend to decline over time. Consumer-<em>b</em>ased weara<em>b</em>le activity trackers that allow users to o<em>b</em>jectively monitor activity levels are now widely availa<em>b</em>le and may offer an a<em>lt</em>ernative method for assisting individuals to remain physically active.</A<em>b</em>stractText><A<em>b</em>stractText>This review aimed to determine the effects of interventions utilizing consumer-<em>b</em>ased weara<em>b</em>le activity trackers on physical activity participation and sedentary <em>b</em>ehavior when compared with interventions that do not utilize activity tracker feed<em>b</em>ack.</A<em>b</em>stractText><A<em>b</em>stractText>A systematic review was performed searching the following data<em>b</em>ases for studies that included the use of a consumer-<em>b</em>ased weara<em>b</em>le activity tracker to improve physical activity participation: Cochrane Controlled Register of Trials, MEDLINE, Pu<em>b</em>Med, Scopus, We<em>b</em> of Science, Cumulative Index of Nursing and Allied Hea<em>lt</em>h Literature, SPORTDiscus, and Hea<em>lt</em>h Technology Assessments. Controlled trials of adu<em>lt</em>s comparing the use of a consumer-<em>b</em>ased weara<em>b</em>le activity tracker with other nonactivity tracker-<em>b</em>ased interventions were included. The main outcome measures were physical activity participation and sedentary <em>b</em>ehavior. All studies were assessed for risk of <em>b</em>ias, and the Grades of Recommendation, Assessment, Development, and Evaluation system was used to rank the quality of evidence. The guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement were followed. A random-effects meta-analysis was completed on the included outcome measures to estimate the treatment effect of interventions that included an activity tracker compared with a control group.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>There was a significant increase in daily step count (standardized mean difference [SMD] 0.24; 95% CI 0.16 to 0.33; P&<em>lt</em>;.001), moderate and vigorous physical activity (SMD 0.27; 95% CI 0.15 to 0.39; P&<em>lt</em>;.001), and energy expenditure (SMD 0.28; 95% CI 0.03 to 0.54; P=.03) and a nonsignificant decrease in sedentary <em>b</em>ehavior (SMD -0.20; 95% CI -0.43 to 0.03; P=.08) following the intervention versus control comparator across all studies in the meta-analyses. In general, included studies were at low risk of <em>b</em>ias, except for performance <em>b</em>ias. Heterogeneity varied across the included meta-analyses ranging from low (I²=3%) for daily step count through to high (I²=67%) for sedentary <em>b</em>ehavior.</p><A<em>b</em>stractText>Utilizing a consumer-<em>b</em>ased weara<em>b</em>le activity tracker as either the primary component of an intervention or as part of a <em>b</em>roader physical activity intervention has the potential to increase physical activity participation. As the effects of physical activity interventions are often short term, the inclusion of a consumer-<em>b</em>ased weara<em>b</em>le activity tracker may provide an effective tool to assist hea<em>lt</em>h professionals to provide ongoing monitoring and support.</A<em>b</em>stractText>

Background: The novel coronavirus (COVID-19) was recently declared a pandemic by the World Health Organization (WHO). The first confirmed case in Saudi Arabia was announced on March 2, 2020. Several psychiatric manifestations may appear during pandemics, especially among frontline healthcare providers.

Objectives: This study sought to explore depression and anxiety levels among healthcare providers during the COVID-19 outbreak in Saudi Arabia.

Methods: This was a cross-sectional study of a convenience sample of 502 healthcare providers in the Ministry of Health. Depression and anxiety were assessed via the Patient Health Questionnaire (PHQ-9) and Generalized Anxiety Disorder 7 (GAD-7) questionnaires, respectively.

Results: The respondents represented various healthcare occupations: administrators (28.49%), nurses (26.29%), physicians (22.11%), non-physician specialists (13.94%), technicians (6.77%), and pharmacists (2.30%). The majority of them were male (68.1%). More than half of them had depressive disorder (55.2%), which ranged from mild (24.9%), moderate (14.5%), and moderately severe (10%) to severe (5.8%). Half of the sample had generalized anxiety disorder (51.4%), which ranged from mild (25.1%) and moderate (11%) to severe (15.3%). Multivariate analysis showed that males were significantly less predicted to have anxiety (Beta=-0.22, P-value &lt;0.04), 30-39 years age group were significantly more predicted to have depression and anxiety group (Beta=0.204, P-value &lt;0.001 and beta=0.521, P-value &lt;0.003 respectively), and nurses had significantly higher mean score of anxiety (Beta=0.445, P-value &lt;0.026).

Conclusions: This study revealed that depression and anxiety are prevailing conditions among healthcare providers. Although efforts were accelerated to support their psychological well-being, more attention should be paid to the mental health of female, 30-39 age group and nursing staff. Promoting healthcare service as a humanitarian and national duty may contribute to making it a more meaningful experience in addition to advocating for solidarity, altruism, and social inclusion. Longitudinal research studies need to be conducted to follow up on healthcare providers' mental health symptoms and develop evidence-based interventions.

Keywords: Anxiety; COVID-19; Depression; Healthcare providers; Pandemic.

<A<em>b</em>stractText>To depict the genomic landscape of Chinese early-stage lung squamous cell carcinoma (LUSC) and investigate its correlation with tumor mutation <em>b</em>urden (TMB), PD-L1 expression, and immune infi<em>lt</em>rates.</A<em>b</em>stractText><p><div>(<em>b</em>)METHODS</<em>b</em>)</div>Whole-exome sequencing was performed on 189 surgically resected LUSC. TMB was defined as the sum of nonsynonymous single nucleotide and indel variants. CD8⁺ tumor-infi<em>lt</em>rating lymphocyte (TIL) density and PD-L1 expression were evaluated <em>b</em>y immunohistochemistry. Six immune infi<em>lt</em>rates were estimated using an online data<em>b</em>ase.</p><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>The median TMB was 9.43 mutations per mega<em>b</em>ase. Positive PD-L1 expression and CD8⁺ TILs density were identified in 24.3% and 78.8%. PIK3CA amplification was associated with significantly higher TMB (P = 0.036). Frequent genetic a<em>lt</em>erations had no impact on PD-L1 expression <em>b</em>ut PIK3CA amplification and KEAP1 mutation were independently associated with significantly lower CD8⁺ TIL density (P &<em>lt</em>; 0.001, P = 0.005, respectively). Low TMB and high CD8⁺ TIL density were independently associated with longer disease-free survival (DFS) while none of them could individually predict the overall survival (OS). Com<em>b</em>ination of TMB and PD-L1 expression or TMB and CD8⁺ TIL density could stratify total populations into two groups with distinct prognosis. Classifying tumor-immune microenvironment <em>b</em>ased on PD-L1 expression and CD8⁺ TIL density showed discrepant genomic a<em>lt</em>erations <em>b</em>ut similar TMB, clinical features, and OS. Nota<em>b</em>ly, patients with different smoking status had distinct prognostic factors.</p><A<em>b</em>stractText>The com<em>b</em>ination of TMB, PD-L1 expression, immune infi<em>lt</em>rates, and smoking status showed the feasi<em>b</em>ility to su<em>b</em>group stratification in Chinese patients with early-stage LUSC, which might <em>b</em>e helpful for future design of personalized immunotherapy trials in LUSC.</A<em>b</em>stractText>

The enterotoxin regions of the heat-labile and heat-stable enterotoxin (LT+ ST+) plasmid, pJY11, originating in a clinically isolated Escherichia coli strain, have been isolated as various-sized deoxyribonucleic acid (DNA) fragments by using cloning vehicles. The structure of the LT+ region and its neighboring DNA regions was studied by utilizing these recombinant plasmids. The LT+ region consisted of at least two genes, toxA and toxB, which could complement each other in trans. The toxA- and toxB-encoded polypeptides (LT subunits A and B, respectively) were identified by their immunological cross-reactivity with Vibrio cholerae enterotoxin subunit A or B. These tox genes and the promoter(s) were localized with respect to the restriction endonuclease cleavage map. The LT+ region was flanked by repeated DNA sequences (designated as beta). Another tox gen(s), encoding ST (designated as toxS), which was also flanked by inverted, repeated DNA sequences (designated as alpha), was located between one of the beta sequences and the LT+ region. These novel DNA structures (beta-alpha-toxS-alpha-toxA-toxB-beta) suggest the possibility that the LT+ region is on a transposon containing an ST transposon within the structure.

It was shown earlier that immune responses against cholera toxin (CT) as well as Vibrio cholerae lipopolysaccharide (LPS) or whole bacterial cells (WC) were protective and that these different antibody specificities co-operated synergistically for protection against experimental cholera. Similarly, antibodies against the heat-labile toxin (LT) and major colonization factors (CFs) of enterotoxingenic Escherichia coli (ETEC) co-operated synergistically for protection against LT-producing ETEC expressing homologous CFs. Studies in humans revealed that repeated oral antigen administration was optimal in inducing intestinal immune responses. Based on these findings oral inactivated vaccines consisting of toxin antigen and whole cells, i.e. the licensed recombinant cholera B subunit (rCTB)-WC cholera vaccine Dukoral®, and candidate ETEC vaccines have been developed. In different trials the rCTB-WC cholera vaccine has provided very high (85-100%) short term protection, which was significantly higher than that induced by the WC component alone, whereas rCTB-WC and WC alone provided comparable (50-60%), long term protection. An oral ETEC vaccine consisting of rCTB and formalin-inactivated E. coli bacteria expressing major CFs was shown to be safe and immunogenic in adults and children in different countries. The vaccine also induced significant protection against non-mild ETEC diarrhoea, i.e. diarrhoea interfering with daily activity in American travellers but not against ETEC diarrhoea in young children in Egypt. Against this background, a modified ETEC vaccine consisting of recombinant E. coli strains overexpressing the major CFs and a more LT like hybrid toxoid (LCTBA) has been developed. This vaccine will be tested soon alone and together with a mucosal adjuvant, i.e. dmLT, in clinical trials.

Forty-four monoclonal antibodies (MAbs) prepared against heat-labile enterotoxins (LTs) from human (LTh) or porcine (LTp) E. coli isolates were characterised, especially with regard to their reactivity with epitopes shared with the heterologous LT and/or cholera toxin (CT), and their toxin neutralising activity. Of 24 MAbs against LTh (all directed against the B subunit portion) 12 cross-reacted with LTp and CT, 4 with LTp but not CT, and 1 with CT but not LTp; 7 MAbs reacted with LTh epitope(s) not shared by either LTp or CT. Among 20 MAbs against LTp (9 directed against the B subunits and 11 against the A subunit) 2 cross-reacted with LTh as well as CT, 13 with LTh but not CT, and 5 MAbs were specific for LTp. Irrespective of whether the anti-LT MAbs were directed against shared or unshared epitopes, or against the A or B subunits, they neutralised their homologous toxin in direct proportion to their toxin-binding titre. The results show how minute differences in enterotoxin primary structures e.g., the LTh and LTp B chains differ in only 4 of 103 amino acid residues, are associated with antigenic epitopes against which toxin-differentiating MAbs with neutralising activity can be produced. Such MAbs are promising tools for species-specific diagnostic detection of enterotoxins in clinical specimens.

Auxiliary liver transplantation (LT) is a special procedure of LT which could be proposed to patients with fulminant hepatic failure (FHF) and has for aim that complete regeneration of the native liver (NL) left in place will allow the graft recipient to resume normal liver function after allograft withdrawal. We report 30 cases of auxiliary LT performed for FHF in 12 European centers. Twenty-five of 30 patients were younger than 50 years. The cause of FHF was hepatitis A virus (HAV) in 4 patients, hepatitis B virus (HBV) in 7, paracetamol overdose in 5, ecstasy in 2, hepatotoxic drugs in 4, autoimmune hepatitis in 2, liver lesions of preeclampsia in 1 and unknown in 5. A postoperative, both clinical and histological follow-up of more than 3 weeks was obtained in 22 patients, enabling us to look for indicators predictive of NL regeneration and outcome. Histological changes observed in the NL included complete regeneration in 68%, incomplete regeneration with obvious fibrous sequelae in 14% and severe liver fibrosis or cirrhosis in 18%, of the 22 patients studied. The percentage and distribution of necrosis observed in tissue samples of the NL at the time of transplantation was not related to the final outcome. Complete NL regeneration was observed in 15 patients, out of whom 14 were younger than 40 years. Patients with complete regeneration were mainly affected by FHF due to HAV, HBV, or paracetamol overdose. After a follow-up of 18/11 (mean/median) months (range, 3 to 67 months), 19 of the 30 patients (63%) survived and 13 of them (68%), i.e., 43% of the 30 patients, had resumed normal NL function, with interrupted immunosuppression, the ultimate goal of emergency auxiliary LT. We conclude that, in patients with FHF, auxiliary LT is a procedure feasible in a number of centers and is associated with a complete regeneration capability of the NL in a majority of survivors, especially in those younger than 40 years. Confirmation of these encouraging preliminary results by large-scale prospective studies is required.

NK cells play a key role in host defense against the beta-herpesvirus CMV through perforin-dependent cytolysis. In this study, we show that human NK cells can also control human CMV (HCMV) infection by a noncytolytic mechanism involving induction of IFN-beta in the virus-infected cell. Both IL-2-activated primary NK cells and an IL-2-dependent NK cell line (NK-92) exhibited potent, noncytolytic anti-HCMV activity at very low E:T cell ratios (<0.1:1). Activated NK cells expressed lymphotoxin (LT)alphabeta on their cell surface, and secreted LTalpha and TNF, all of which contributed to the NF-kappaB-dependent release of IFN-beta from infected fibroblasts. IFN-beta produced by fibroblasts and NK cell-produced IFN-gamma combined to inhibit HCMV replication after immediate early gene expression. These results highlight an efficient mechanism used by NK cells to activate IFN-beta expression in the infected target cell that contributes to the arrest of virion production and virus spread without cellular elimination.

The S. typhimurium strain (TML deltaaroC deltassaV) WT05, harbouring defined deletions in genes involved in both the aromatic biosynthesis pathway (aroC) and the Salmonella Pathogenicity Island-2 (SPI-2) (ssaV) was shown to be significantly attenuated in C57 BL/6 interferon gamma knockout mice following oral inoculation. Similarly, the S. typhi strain (Ty2 deltaaroC deltassaV) ZH9 harbouring the aroC and ssaV mutations propagated less efficiently than wild type in human macrophages. These studies demonstrated the attractive safety profile of the aroC ssaV mutant combination. Strains S. typhimurium (TML deltaaroC deltassaV ) WT05 and S. typhi (Ty2 deltaaroC deltassaV) ZH9 were subsequently tested as vaccine vectors to deliver E. coli heat-labile toxin (LT-B) mucosally to mice. Mice inoculated orally with S. typhimurium (TML deltaaroC deltassaV) WT05 expressing LT-B (WT05/LT-B) elicited high titres of both LT-specific serum IgG and intestinal IgA, although no specific IgA was detected in the vagina. Similarly, intranasal inoculation of mice with S. typhi (Ty2 deltaaroC deltassaV) ZH9 expressing LT-B (ZH9/LT-B) elicited even higher titres of LT-specific serum antibody as well as LT-specific Ig in the vagina. We conclude that deltaaroC deltassaV strains of Salmonella are highly attenuated and are promising candidates both as human typhoid vaccines and as vaccine vectors for the delivery of heterologous antigens.

The specificity of the pathway used by Vibrio cholerae for extracellular transport of cholera toxin (CT) and other proteins was examined in several different ways. First, V. cholerae was tested for the ability to secrete the B polypeptides of the type II heat-labile enterotoxins of Escherichia coli. Genes encoding the B polypeptide of LT-IIb in pBluescriptKS- phagemids were introduced into V. cholerae by electroporation. Culture supernatants and periplasmic extracts were collected from cultures of the V. cholerae transformants, and the enterotoxin B subunits were measured by an enzyme-linked immunosorbent assay. Results confirmed that the B polypeptides of both LT-IIa and LT-IIb were secreted by V. cholerae with efficiencies comparable to that measured for secretion of CT. Second, the plasmid clones were introduced into strain M14, an epsE mutant of V. cholerae. M14 failed to transport the B polypeptides of LT-IIa and LT-IIb to the extracellular medium, demonstrating that secretion of type II enterotoxins by V. cholerae proceeds by the same pathway used for extracellular transport of CT. These data suggest that an extracellular transport signal recognized by the secretory machinery of V. cholerae is present in LT-IIa and LT-IIb. Furthermore, since the B polypeptide of CT has little, if any, primary amino acid sequence homology with the B polypeptide of LT-IIa or LT-IIb, the transport signal is likely to be a conformation-dependent motif. Third, a mutant of the B subunit of CT (CT-B) with lysine substituted for glutamate at amino acid position 11 was shown to be secreted poorly by V. cholerae, although it exhibited immunoreactivity and ganglioside GM1-binding activity comparable to that of wild-type CT-B. These findings suggest that Glu-11 may be within or near the extracellular transport motif of CT-B. Finally, the genetic lesion in the epsE allele of V. cholerae M14 was determined by nucleotide sequence analysis.