Alternate-Day Fasting Ameliorates Newly Established Sjögren’s Syndrome-like Sialadenitis in Non-Obese Diabetic Mice

Intermittent fasting confers protections to various diseases including autoimmune disorders, but the specific effects of intermittent fasting on Sjögren’s syndrome (SS) remains inconclusive. The present study was undertaken to determine the specific impact of alternate-day fasting (ADF) on newly established SS-like sialadenitis using non-obese diabetic (NOD) mice. Female NOD mice were deprived of food every other day from 10 to 13 weeks of age, the early stage of established SS, and then analyzed for the disease characteristics. Mice in the ADF group had higher salivary flow rate and attenuated submandibular gland (SMG) inflammation, compared to the control mice fed with standard chow ad libitum. The improvements were accompanied with a decrease in the total leukocytes, T and B lymphocytes and activated CD4 and CD8 T cells, and a down-regulation of pro-inflammatory cytokines IFN-γ and IL-17, chemokine receptor CXCR3 and its ligands CXCL9 and CXCL11 in the SMGs. ADF also led to elevated mRNA levels of water channel protein aquaporin 5 and tight junction protein claudin-1, two factors crucial for normal salivary secretion in the SMGs. In addition, ADF reduced the proportion of IFN-γ- and IL-17- expressing CD4 T cells and diminished mRNA levels of IFN-γ, TNF-α, and IL-17 in the total submandibular draining lymph node cells. Taken together, ADF is effective in ameliorating newly established SS-associated salivary gland exocrinopathy in NOD mice.     

Immunomodulatory effect of mushrooms on cytotoxic activity and cytokine production of intestinal lamina propria leukocytes does not necessarily depend on β-glucan contents

We evaluated the effects of seven mushroom extracts (Grifola frondosa, Pholiota nameko, Panellus serotinus, Hypsizygus marmoreus, Pleurotus cornucopiae, Armillaria mellea, and Flammulina velutipes) on cytotoxic activity and cytokine production of lamina propria leukocytes (LPLs) isolated from rat small (S) and large (L) intestinal mucosa. Boiling water extracts from seven species of mushrooms showed no direct cytotoxicity against the YAC-1 target cells. However, prominent increases of cytotoxicity were observed in S- and L-LPLs co-cultured with P. serotinus extract. Cytokine production (TNFα, IFNγ, IL-12 p70, and IL-4) of S- and L-LPLs was stimulated in response to P. cornucopiae extract. Mushroom extracts contributed to target cell adhesion and/or cytokine production in the effector cells. The promotion of cytotoxic activity in S- and L-LPLs was not necessarily related to β-glucan content of the mushroom.     

Eosinophil-Derived Neurotoxin (EDN/RNase 2) and the Mouse Eosinophil-Associated RNases (mEars): Expanding Roles in Promoting Host Defense

The eosinophil-derived neurotoxin (EDN/RNase2) and its divergent orthologs, the mouse eosinophil-associated RNases (mEars), are prominent secretory proteins of eosinophilic leukocytes and are all members of the larger family of RNase A-type ribonucleases. While EDN has broad antiviral activity, targeting RNA viruses via mechanisms that may require enzymatic activity, more recent studies have elucidated how these RNases may generate host defense via roles in promoting leukocyte activation, maturation, and chemotaxis. This review provides an update on recent discoveries, and highlights the versatility of this family in promoting innate immunity.     

Differential leukocyte count method for bovine low somatic cell count milk

Whereas many differential leukocyte count methods for high somatic cell count (SCC) milk from mastitic cows are available, only a few have been developed for low SCC milk. We have developed a flow cytometric differential leukocyte count method for low SCC milk. The procedure consists of 1) 1.5 ml of diluted milk sample (30%, vol/vol dilution with PBS), 2) centrifugation, 3) leukocyte labeling with SYTO 13 and 4) flow cytometric analysis. Four major leukocyte populations can be clearly identified in the green fluorescence-side scatter dot plot: lymphocytes and monocytes (LM), polymorphonuclear neutrophils (PMN), mature macrophages (Mphi), and cells with apoptotic features based on chromatin condensation and nuclear fragmentation. The optimal processing temperature was 20 degrees C. Significant differences among samples with similar differential leukocyte counts were found. Storage of milk samples during 2 d at 7 degrees C had no effect on differential leukocyte count. Using the new method, differential leukocyte count was performed in low SCC milk samples from cows in early, mid, and late lactation. In accordance with previous studies, PMN and Mphi percentages were lower and LM percentages were higher in early lactation than in the other stages of lactation. The percentage of cells with apoptotic features was higher in early lactation than in mid and late lactation. In conclusion, a rapid, simple, accurate, and reproducible standard procedure was developed to determine the differential leukocyte count (Mphi, PMN, LM, and cells with apoptotic features) of bovine low SCC milk.     

Human Adipose-Derived Mesenchymal Progenitor Cells Engraft into Rabbit Articular Cartilage

Mesenchymal stem cells (MSCs) are known to have the potential for articular cartilage regeneration, and are suggested for the treatment of osteoarthritis (OA). Here, we investigated whether intra-articular injection of xenogeneic human adipose-derived mesenchymal progenitor cells (haMPCs) promoted articular cartilage repair in rabbit OA model and engrafted into rabbit articular cartilage. The haMPCs were cultured in vitro, and phenotypes and differentiation characteristics of cells were evaluated. OA was induced surgically by anterior cruciate ligament transection (ACLT) and medical meniscectomy of knee joints. At six weeks following surgery, hyaluronic acid (HA) or haMPCs was injected into the knee joints, the contralateral knee served as normal control. All animals were sacrificed at the 16th week post-surgery. Assessments were carried out by macroscopic examination, hematoxylin/eosin (HE) and Safranin-O/Fast green stainings and immunohistochemistry. The data showed that haMPC treatment promoted cartilage repair. Signals of human mitochondrial can be directly detected in haMPC treated cartilage. The haMPCs expressed human leukocyte antigen I (HLA-I) but not HLA-II-DR in vivo. These results suggest that intra-articular injection of haMPCs promotes regeneration of articular cartilage in rabbit OA model, and support the notion that MPCs are transplantable between HLA-incompatible individuals.     

对流换热理论在玻璃窑炉换热器设计中的应用

本文主要分析了管内层流对流换热时两种不同情况下努谢尔特准数的数值及在换热器设计中的指导意义。     

Presence of Escherichia coli, Trueperella pyogenes, α-hemolytic streptococci,and coagulase-negative staphylococci and prevalence of subclinical endometritis

Subclinical endometritis (SCE) has a severe negative effect on reproductive performance of dairy cows. Different studies have shown a wide variety of cut points for the polymorphonuclear neutrophilic leukocyte (PMNL) percentage to define SCE and differing sampling times. Possible risk factors for SCE, such as low body condition score, hyperketonemia, and increased haptoglobin concentrations have been demonstrated. The aim of this study was to determine the relationship between bacterial findings in the uterus and PMNL in the early postpartum period and to compare 2 classification schemes for the diagnosis of SCE. Cytological and bacteriological samples from the uterus were taken from 149 cows at 2 consecutive times postpartum [10 ± 1 and 24 ± 1 d in milk (DIM)] and additionally, 131 cows were sampled at 21 to 27 DIM. For sample collection, the cytobrush technique was used and vaginal discharge was examined by vaginoscopy. Polymorphonuclear neutrophilic leukocyte cut points were set at 5, 10, and 18%. Cows positive with α-hemolytic streptococci at 10 ± 1 DIM had significant higher PMNL percentages at 21 to 27 DIM and greater median days to pregnancy (193.5 d) than cows not infected (123.0 d). Cows with Trueperella pyogenes at 10 ± 1 DIM had higher odds (5.2) of having more than 18% PMNL at 21 to 27 DIM compared with cows without infection. Escherichia coli had no effect on the prevalence of SCE regardless of the cut point. Primiparous cows with more than 18% PMNL had greater median days to pregnancy (144.5 d) than cows with less than 5% (80.0 d) or cows between 5 and 18% PMNL (68.0 d). Cows between 5 and 18% PMNL or 10 and 18% PMNL showed the best reproductive performance. The results demonstrate that an early infection with α-hemolytic streptococci increased the PMNL percentage 2 wk later and that 18% PMNL at 21 to 27 DIM was a predictive cut point for primiparous cows to diagnose animals with or without SCE.     

A design of laminated composite plates using graded orthotropic fiber-reinforced composite plies

A new lamination scheme is proposed through the design of a graded orthotropic fiber-reinforced composite ply for achieving continuous variations of material properties along the thickness direction of laminated composite plates. First, a micro-structure of graded unidirectional fiber-reinforced composite ply is designed and its effective graded elastic properties are estimated using finite element procedure. Next, the new lamination scheme is demonstrated through the conversion of a conventional laminated composite plate (CLCP) into a conventional-graded laminated composite plate (CGLCP) utilizing presently designed graded orthotropic composite ply. The suitability of this conversion/proposed lamination scheme is substantiated through the bending analysis of both the plates (CLCP and CGLCP).     

Image correlation microscopy for uniform illumination

Image cross-correlation microscopy is a technique that quantifies the motion of fluorescent features in an image by measuring the temporal autocorrelation function decay in a time-lapse image sequence. Image cross-correlation microscopy has traditionally employed laser-scanning microscopes because the technique emerged as an extension of laser-based fluorescence correlation spectroscopy. In this work, we show that image correlation can also be used to measure fluorescence dynamics in uniform illumination or wide-field imaging systems and we call our new approach uniform illumination image correlation microscopy. Wide-field microscopy is not only a simpler, less expensive imaging modality, but it offers the capability of greater temporal resolution over laser-scanning systems. In traditional laser-scanning image cross-correlation microscopy, lateral mobility is calculated from the temporal de-correlation of an image, where the characteristic length is the illuminating laser beam width. In wide-field microscopy, the diffusion length is defined by the feature size using the spatial autocorrelation function. Correlation function decay in time occurs as an object diffuses from its original position. We show that theoretical and simulated comparisons between Gaussian and uniform features indicate the temporal autocorrelation function depends strongly on particle size and not particle shape. In this report, we establish the relationships between the spatial autocorrelation function feature size, temporal autocorrelation function characteristic time and the diffusion coefficient for uniform illumination image correlation microscopy using analytical, Monte Carlo and experimental validation with particle tracking algorithms. Additionally, we demonstrate uniform illumination image correlation microscopy analysis of adhesion molecule domain aggregation and diffusion on the surface of human neutrophils.     

Validation of an automated cell counter to determine leukocyte differential counts in neonatal Holstein calves

Recent advances in the understanding of risk factors and biomarkers in calves entering rearing facilities show promise for identifying high-risk calves on arrival at veal and dairy beef operations. Rapid automated leukocyte differential cell counts may be a good addition for augmenting or refining calf risk identification on-farm. The objective of this study was to validate an automated leukocyte cell counter, the QScout BLD test (Advanced Animal Diagnostics, Morrisville, NC), for its ability to determine leukocyte differential cell counts in neonatal Holstein calves. From June to July 2018, blood samples collected in EDTA anticoagulant from 235 calves upon arrival at an independent veal research facility in Ontario, Canada, were evaluated using the QScout BLD test and manually by microscopy. We compared these leukocyte differential counts using Lin's concordance correlation coefficient (ρ) and found very good agreement between tests for neutrophil counts (ρ = 0.83); fair agreement for lymphocyte counts (ρ = 0.32); fair agreement for the ratio of neutrophils to lymphocytes (ρ = 0.36); slight agreement for monocyte counts (ρ = 0.14); and slight agreement for eosinophil counts (ρ = 0.20). We further examined test results to determine if they differed in their classification of samples as being above, within, or below reported 95% reference intervals for neonatal Holstein calves. Classification between tests resulted in very good agreement for neutrophils and lymphocytes, with only 4.2% and 5.8% disagreement in classification, respectively. We observed moderate agreement for monocytes, with 23.3% classified differently, and poor agreement for eosinophils, with 70.3% classified differently. Further study is required to determine the role of leukocyte profiling in the risk assessment of calves arriving at calf-rearing facilities.