BLOOD AND ACID-BASE CHANGES IN CAILVES EICPERIMENTALLY INFECTED WITH SALMONELLA TYPHIMPRIUM

The experimental design of this study was included (12) Friesian calves aged between 3-5 weeks. Calves were divided into two groups. Group I consisted of (8) calves were infected experimentally with 1.5 x 10“ of Salmonella typhimurium and often that treatment with chloramphenicol and electrolytes fluid were takes place. Group II consisted of (4) calves were infected with organisms but without treatment. The clinical findings of the disease were characterized by two forms which includes the septicemic and enteric forms. Haematological changes revealed a variable number of the total and differential leukocytic count, increased PCV, fibnnogen and decreased of the total plasma protein. . The changes in the acid-based balance indicate the development of metabolic acidosis.

Immunoglobulin G1 Fc glycosylation as an early hallmark of severe COVID-19

Background Immunoglobulin G1 (IgG1) effector functions are impacted by the structure of fragment crystallizable (Fc) tail-linked N-glycans. Low fucosylation levels on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein specific (anti-S) IgG1 has been described as a hallmark of severe coronavirus disease 2019 (COVID-19) and may lead to activation of macrophages via immune complexes thereby promoting inflammatory responses, altogether suggesting involvement of IgG1 Fc glycosylation modulated immune mechanisms in COVID-19. Methods In this prospective, observational single center cohort study, IgG1 Fc glycosylation was analyzed by liquid chromatography - mass spectrometry following affinity capturing from serial plasma samples of 159 SARS-CoV-2 infected patients. Findings At baseline close to disease onset, anti-S IgG1 glycosylation was highly skewed when compared to total plasma IgG1. A rapid, general reduction in glycosylation skewing was observed during the disease course. Low anti-S IgG1 galactosylation and sialylation as well as high bisection were early hallmarks of disease severity, whilst high galactosylation and sialylation and low bisection were found in patients with low disease severity. In line with these observations, anti-S IgG1 glycosylation correlated with various inflammatory markers. Interpretation Association of low galactosylation, sialylation as well as high bisection with disease severity suggests that Fc-glycan modulated interactions contribute to disease mechanism. Further studies are needed to understand how anti-S IgG1 glycosylation may contributes to disease mechanism and to evaluate its biomarker potential. Funding This project received funding from the European Commission's Horizon2020 research and innovation program for H2020-MSCA-ITN IMforFUTURE, under grant agreement number 721815.

Inferring a causal relationship between ceramide levels and COVID-19 respiratory distress

A causal relationship between plasma ceramide concentration and respiratory distress symptoms in COVID-19 patients is inferred. In this study, plasma samples of 52 individuals infected with COVID-19 were utilized in a lipidomic analysis. Lipids belonging to the ceramide class exhibited a 400-fold increase in total plasma concentration in infected patients. Further analysis led to the demonstration of concentration dependency for severe COVID-19 respiratory symptoms in a subclass of ceramides. The subclasses Cer(d18:0/24:1), Cer(d18:1/24:1), and Cer(d18:1/22:0) were shown to be increased by 48-, 40-, and 33-fold, respectively, in infected plasma samples and to 116-, 91- and 50-fold, respectively, in plasma samples with respiratory distress. Hence, monitoring plasma ceramide concentration, can be a valuable tool for measuring effects of therapies on COVID-19 respiratory distress patients.

Pleiotropic Effects of Atorvastatin Result in a Downregulation of the Carboxypeptidase U System (CPU, TAFIa, CPB2) in a Mouse Model of Advanced Atherosclerosis

Statins (hydroxymethyl-glutaryl-CoA-reductase inhibitors) lower procarboxypeptidase U (proCPU, TAFI, proCPB2). However, it is challenging to prove whether this is a lipid or non-lipid-related pleiotropic effect, since statin treatment decreases cholesterol levels in humans. In apolipoprotein E-deficient mice with a heterozygous mutation in the fibrillin-1 gene (ApoE−/−Fbn1C1039G+/−), a model of advanced atherosclerosis, statins do not lower cholesterol. Consequently, studying cholesterol-independent effects of statins can be achieved more straightforwardly in these mice. Female ApoE −/−Fbn1C1039G+/− mice were fed a Western diet (WD). At week 10 of WD, mice were divided into a WD group (receiving WD only) and a WD + atorvastatin group (receiving 10 mg/kg/day atorvastatin +WD) group. After 15 weeks, blood was collected from the retro-orbital plexus, and the mice were sacrificed. Total plasma cholesterol and C-reactive protein (CRP) were measured with commercially available kits. Plasma proCPU levels were determined with an activity-based assay. Total plasma cholesterol levels were not significantly different between both groups, while proCPU levels were significantly lower in the WD + atorvastatin group. Interestingly proCPU levels correlated with CRP and circulating monocytes. In conclusion, our results confirm that atorvastatin downregulates proCPU levels in ApoE−/−Fbn1C1039G+/− mice on a WD, and evidence was provided that this downregulation is a pleiotropic effect of atorvastatin treatment.

PSIX-13 Activation of cell signaling pathways by galectins in cow blood

Galectins are part of a conserved family of β-galactoside-binding proteins that contribute to critical biological events during mammalian gestation and increasingly recognized for a possible role in the immune response of the cow. The objective of this study was to evaluate the effect of Galectins on signal transduction and cell activation in cow blood. Whole blood was collected aseptically from the jugular vein of healthy Holstein Friesian dairy cows (N=3). Blood samples (2.5ml) in duplicate were treated with 150µl of the four different type of recombinant galectins (1, 3, 4, and 9) respectively and untreated samples were served as control. The concentration of total plasma protein was determined using the Pierce BCA kit. Protein expression profiling was performed using1,358 antibodies on the Full Moon BioSystems’ Signaling Explorer antibody array covering 20 cell signaling pathways, as recommended by the manufacturer using an Agilent microarray scanner. Data normalization was performed using GeneSpring GX software to generate fold changes in gene expression and then filtered to obtain a list of significantly upregulated and downregulated genes. Features were extracted from protein array images of samples treated with Galectin 1, 3, 4, 9, and untreated sample as a control group. Treatment with all four Galectins increased the concentration of total plasma protein. Average increases due to treatment with Gal1, Gal3, Gal4, and Gal9 were 27%, 10%, 20%, and 14% respectively. ANOVA test showed significance difference among the groups (p < 0.05). Dunnett option was used to compare each of the treated samples to the control group as a baseline. The results also showed that there was significant difference between the control group and any of the treated group (p < 0.05). Distinct signaling pathways are activated in response to Galectin exposure. Further studies are needed to define their regulation and functional impact on cow health.

Effect of Physiological Status And Parity On Metabolic And Trace Elements Profile of Crossbred Rambouillet Sheep of Himalayan Region

The study was designed to evaluate the effect of physiological status and parity on metabolic profile in crossbred Rambouillet ewes of Himalyan region. The study was conducted on 20 ewes divided into two groups, primiparous (PP) and multiparous (MP) with 10 ewes in each group. Blood samples were collected on 4- and 1-week pre-lambing and 1- and 4-weeks post-lambing to measure metabolic parameters and minerals. The glucose(p<0.01), Total plasma protein (TPP) (p<0.05), albumin (p<0.05), BUN (p<0.05), cholesterol (p<0.05), triglyceride (p<0.01), HDL-C (P<0.05), Calcium (p<0.01), Phosphours (p<0.05), Magnesium (p<0.01), Cu (p<0.05), and Zn (p<0.01) levels revealed significant change along the time with the concentration decreasing 3 weeks pre lambing to immediate post lambing thereafter levels increased steadily. Significant increase (P<0.01) was observed in NEFA, AST, GGT, Iron(P<0.05), and bilirubin (P<0.05) concentrations along the sampling time. No group difference was observed in any of the parameters however, parity and time interaction was observed in Glucose, NEFA, GGT, calcium, and phosphorus. While NEFA levels were significantly high in pre-lambing in PP ewes compared to MP ewes, the post-lambing levels were significantly high in MP ewes. Pre-lambing levels of GGT were at par between the two groups; however, post-lambing levels were significantly high in MP ewes. Glucose, Ca and Pi were low during pre-lambing in PP ewes and post-lambing in MP ewes. The result showed that ewes shows a significant change in metabolic profile and trace minerals during late gestation and immediate post-partum, however these changes were more pronounced during late gestation in primiparous and post lambing in multiparous.

A Novel Microfluidic Chip for Fast, Sensitive Quantification of Plasma Extracellular Vesicles as Biomarkers in Patients With Osteosarcoma

Plasma circulating extracellular vesicle (EV) has emerged as a promising biomarker for diagnosis and prognosis of various epithelial tumors. However, fast and efficient capture of EVs with microfluidic chip in sarcoma remains to be established. Herein, we reported a ZnO-nanorods integrated (ZNI) microfluidic chip, where EV capture antibody was uniformly grafted to the surface of the ZnO-nanorods of the chip to enhance the plasma turbulence formation and the capture efficiency at the micro-scale. Based on osteosarcoma (OS) cell line, we demonstrated that a combination of CD81 and CD63 antibody on ZNI chip yielded the greatest amount of total EVs, with an extra sensitive limit of detection (LOD) of ~104 particles mL-1. Furthermore, the addition of fluorescent labeling of Vimentin (VIM), a previously reported sarcoma cell surface biomarker, could enabled the dual visualization of total plasma EVs and VIM-positive EVs from OS patients’ plasma. Based on our ZNI chip, we found that the amount of plasma total EVs was significantly different between OS and healthy donors (1562 a.u. versus 639 a.u., p&lt; 0.05), but not between metastatic and nonmetastatic OS (p&gt; 0.05). Interestingly, patients with metastatic disease had a significantly greater amount of VIM-positive EVs (1411 a.u. versus 231 a.u.., p&lt; 0.05) and increased VIM-positive/total EVs ratio (0.943 versus 0.211, p&lt; 0.05) in comparison with the nonmetastatic counterpart. Therefore, our ZNI microfluidic chip has great potential for the fast quantification of plasma EVs, and the microfluidic-based quantification of total and VIM-positive EVs might serve as a promising biomarker for the diagnosis and surveillance in OS patients.

Dynamic change of blood profile in rat models with acute skin injury artificially infected with methicillin-resistant Staphylococcus aureus

Background and Aim: A wound is a common problem for humans and animals. The wound becomes more severe if it is infected by bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA). The wound healing mechanism involves various factors, either in the local tissue or the bloodstream. However, the presentation of infected wound healing regarding its impacts on the dynamic change of blood profile is not clearly understood. This study aimed to explore the impacts of wound creation on the blood profile in rat models with and without being artificially infected by MRSA. Materials and Methods: Thirty male Sprague-Dawley rats (6 months old; weight, 300 g) were used as the model. They were divided into three groups: Without wound creation (C), wounded without infection (CW), and wounded and artificially infected by MRSA (CWI). Groups CW and CWI were shaved and induced with 4 mm two-round full-thickness biopsy on the back. Furthermore, group CWI was artificially infected by 105 colony-forming units of MRSA. The blood samples were collected through the tail vein from days 1 to 5. The blood parameters included blood profile, total plasma protein, C-reactive protein, CD4+, CD8+, CD4+/CD8+, and COX-2. The data were analyzed using the Statistical Package for the Social Sciences, version 16 (SPSS, IBM, Armonk, NY, USA). Results: The result showed that the presentation of a wound with and without MRSA infection significantly changed the total erythrocytes, leukocytes, neutrophils, lymphocytes, total plasma protein, C-reactive protein, and the subset of circulatory CD4+, CD8+, and COX-2 (p≤0.05). In addition, the wound infected with MRSA impacts the mean corpuscular volume (p≤0.05). Conclusion: Moreover, the presentation of the wound with and without MRSA infection induces dynamic changes on various blood profile parameters.

Plasma Sphingomyelins in Late-Onset Alzheimer’s Disease

Background: Altered plasma levels of sphingolipids, including sphingomyelins (SM), have been found in mouse models of Alzheimer’s disease (AD) and in AD patient plasma samples. Objective: This study assesses fourteen plasma SM species in a late-onset AD (LOAD) patient cohort (n = 138). Methods: Specimens from control, preclinical, and symptomatic subjects were analyzed using targeted mass-spectrometry-based metabolomic methods. Results: Total plasma SM levels were not significantly affected by age or cognitive status. However, one metabolite that has been elevated in manifest AD in several recent studies, SM OHC14:1, was reduced significantly in pre-clinical AD and MCI relative to normal controls. Conclusion: We recommend additional comprehensive plasma lipidomics in experimental and clinical biospecimens related to LOAD that might advance the utility of plasma sphingomyelin levels in molecular phenotyping and interpretations of pathobiological mechanisms.