Postischemic Neuroprotection Associated With Anti-Inflammatory Effects by Mesenchymal Stromal Cell-Derived Small Extracellular Vesicles in Aged Mice

Background and Purpose: Small extracellular vesicles (sEVs) obtained from mesenchymal stromal cells (MSCs) were shown to induce ischemic neuroprotection in mice by modulating the brain infiltration of leukocytes and, specifically polymorphonuclear neutrophils. So far, effects of MSC-sEVs were only studied in young ischemic rodents. We herein examined the effects of MSC-sEVs in aged mice. Methods: Male and female C57Bl6/j mice (8–10 weeks or 15–24 months) were exposed to transient intraluminal middle cerebral artery occlusion. Vehicle or sEVs (equivalent of 2×10 6 MSCs) were intravenously administered. Neurological deficits, ischemic injury, blood-brain barrier integrity, brain leukocyte infiltration, and blood leukocyte responses were evaluated over up to 7 days. Results: MSC-sEV delivery reduced neurological deficits, infarct volume, brain edema, and neuronal injury in young and aged mice of both sexes, when delivered immediately postreperfusion or with 6 hours delay. MSC-sEVs decreased leukocyte and specifically polymorphonuclear neutrophil, monocyte, and macrophage infiltrates in ischemic brains of aged mice. In peripheral blood, the number of monocytes and activated T cells was significantly reduced by MSC-sEVs. Conclusions: MSC-sEVs induce postischemic neuroprotection and anti-inflammation in aged mice.

Identification of Persons Who Are Responsive To Wood Smoke Particle-Induced Airway Inflammation With Assessment of The Effect of GSTM1, Asthma Status And Sex On This Response.

Background: We are currently screening human volunteers to determine their sputum polymorphonuclear neutrophil (PMN) response 6 and 24 hours following initiation of exposure to wood smoke particles (WSP). Inflammatory responders (>10% increase in %PMN) are identified for their subsequent participation in mitigation studies against WSP-induced airways inflammation. In this report we compared responder status (N=52) at both 6 and 24hr time points to refine/expand its classification, assessed the impact of the GSTM1 genotype, asthma status and sex on responder status, and explored whether sputum soluble phase markers of inflammation correlate with PMN responsiveness to WSP. Results: In the entire cohort, we found a significant, but very small, decrease in FVC and systolic blood pressure immediately following WSP exposure and sputum %PMNs were significantly increased at 24 hours post exposure, the latter finding was also significantly correlated with sputum IL-1b, IL-6, IL-8, and PMN/mg; a similar response was not found at the 6 hour %PMN response. Blood endpoints in the entire cohort showed a significant increase in %PMN and PMN/mg at 6 but not 24 hours. Six-hour responders tended to be 24-hour responders and vice versa, but 24-hour responders also had significantly increased IL-1b, IL-6, IL-8 at 24 hours post WSP exposure. The GSTM1 null genotype significantly (p<0.05) enhanced the %PMN response at 6 hours in the entire cohort, by 24% in the 24-hour responders and not at all in the 6 hours responders. Asthma status enhanced the 24 hour %PMN response in the entire cohort and in the 6- and 24-hour responders. Sex had no effect on %PMN response. Conclusions: The 24 hour time point was more informative than the 6 hour time point in optimally defining airway inflammatory responsiveness to WSP exposure. GSTM1 and asthma status are significant effect modifiers of this response. These study design and subject parameters should be considered before enrolling volunteers for proof-of-concept WSP mitigation studies.

256 Effects of Transition Period Nutrition on Uterine Health

The transition from gestation to lactation, also known as the transition period, is a critical time for dairy cows. This phase is typically defined as 3 wk before parturition through 3 wk after parturition. Peak disease incidence (shortly after parturition) corresponds with the time of greatest negative energy balance (NEB), the peak in blood concentrations of nonesterified fatty acids, and the greatest acceleration of milk yield. Decreased fertility in the face of increasing milk production may be attributable to greater severity of postpartal NEB resulting from inadequate transition management or increased rates of disease. The depth and duration of NEB is highly related to dry matter intake. Formulation and delivery of appropriate diets that limit total energy intake to requirements but also provide proper intakes of all other nutrients (including the most limiting amino acids Met and Lys) before calving can help lessen the extent of NEB after calving. Cows that received controlled-energy diets during the last 3 wk prepartum had shorter days to pregnancy than cows that consumed high-energy diets in this time period. Additionally, a fully acidified prepartum diet formulated with a higher Ca concentration improved reproductive performance and uterine immune function in the postpartum period through the decrease of days to first ovulation, a tendency to decrease service per conception rate, improved glandular morphology, a tendency to increase polymorphonuclear neutrophil infiltration, and a tendency to increase the tight-junction protein occludin. Supplementation of rumen-protected methionine during the transition period resulted in improved uterine immune function through improved glandular morphology, increased neutrophil infiltration number after calving, and discovery of neutrophil extracellular trap formation in bovine endometrial tissue. In conclusion, nutritional strategies during the transition impact uterine health and fertility in dairy cows.

Clinical utility of Neutrophil Lymphocyte Ratio (NLR) as a marker of Spontaneous Bacterial Peritonitis (SBP) in patients with cirrhosis-An exploratory study

Background and objective: The diagnosis of spontaneous bacterial peritonitis (SBP) is made by presence of ≥250 polymorphonuclear neutrophil (PMN)/mm3 in the ascitic fluid. Paracentesis despite being the gold standard has its inherent risks and complications. Blood neutrophil-lymphocyte ratio is a simple test for inflammation. Highly sensitive C reactive protein (hsCRP) is a marker of inflammation which is mainly synthesized by the liver. We aimed to evaluate clinical utility of NLR and hsCRP as less invasive tests for diagnosis of SBP. Methods: Fifty cases of cirrhosis with ascites with SBP and 50 age and sex matched controls of cirrhosis with ascites without SBP were enrolled for the study. NLR was calculated and hsCRP value was determined in both the groups and compared using independent t test. The sensitivity and specificity of NLR was estimated as a test for SBP diagnosis by using receiver operator characteristics (ROC) curve. Results: NLR was found to be significantly higher in SBP patients (6.75 +/- 2.7) than those without it (2.81+/-1.06) with p value < 0.01. hsCRP was raised in both groups, 18.93+/-5.00 and 17.46+/-6.19 in cases and controls respectively, but there was no statistical difference between the two groups. For SBP diagnosis, a blood NLR > 3.38 had a sensitivity of 94% and a specificity of 80%. Interpretation and conclusions: NLR could be used as a novel and less invasive test for diagnosis of SBP. hsCRP has a blunted rise in patients with cirrhosis with SBP and cannot be used as diagnostic marker.

Effect of BLV Infection on the Immune Function of Polymorphonuclear Neutrophil in Dairy Cows

Enzootic bovine leukemia is a late-onset, neoplastic infection caused by the bovine leukemia virus (BLV). BLV infection hinders the function of the immune system and induces other diseases, which negatively affects the performance and health of the infected cows. As the first line of defense against invading foreign pathogenic microorganisms, polymorphonuclear neutrophil (PMN) plays a vital role in the immune system of dairy cows. However, research on the effect of BLV infection on the immune function of PMN in dairy cows is scarce. Therefore, this experiment aimed to elucidate the effects and effect mechanisms of BLV infection on the immune function of PMN in dairy cows with different BLV provirus loads by detecting the chemotaxis, migration, adhesion, phagocytosis, respiratory burst function, and the formation of NETs. The experimental results showed that BLV infection had no significant effect on the phagocytosis of PMN but inhibited their migration and respiratory burst function, and the effects were closely related to the BLV provirus load. Under high BLV provirus load, PMN produced large amounts of NETs, chemokine CXCL7, adhesion molecule CD18, and pro-inflammatory factors IL-8 and TNF-α, triggering inflammatory responses, and tissue damage. The results of this study will help reveal the reason why BLV infection causes the high incidence of mammary gland inflammation in dairy cows.

Fecal Lactoferrin and Other Putative Fecal Biomarkers in Crohn’s Disease: Do They Still Have a Potential Clinical Role?

<b><i>Introduction:</i></b> The need for noninvasive markers of disease activity is mandatory in the assessment of Crohn’s disease (CD). The most widely fecal biomarker in CD, despite several limits, is fecal calprotectin. This review aims to elucidate the role, if any, of all other fecal biomarkers, as alternative tools for assessing clinical and endoscopic disease activity, and predict capsule endoscopy findings, response to therapy, disease relapse, and postoperative recurrence. These fecal biomarkers included lactoferrin, S100A12, high mobility group box 1, neopterin, polymorphonuclear neutrophil elastase, fecal hemoglobin, alpha1-antitrypsin, lysozyme, human beta-defensin-2, neutrophil gelatinase-associated lipocalin, matrix metalloproteinase-9, chitinase 3-like-1, M2-pyruvate kinase, myeloperoxidase, and eosinophil proteins. <b><i>Methods:</i></b> A systematic electronic search in the medical literature was performed up to April 2020. Seventy eligible studies were identified out of 859 citations. Data were grouped according to the assessment of clinical and endoscopic disease activity, capsule endoscopy findings, response to therapy, prediction of relapse, and postoperative recurrence. <b><i>Results:</i></b> The overall correlation between lactoferrin and clinical indexes is poor, while performance is good with endoscopic scores. Lactoferrin seems to represent a reasonably good surrogate marker of response to therapy and to be potentially useful in identifying patients at high risk for endoscopic relapse or postoperative recurrence. The evaluation of the performance of all other fecal markers is limited by the lack of adequate data. <b><i>Conclusions:</i></b> None of the fecal markers so far represents an acceptable alternative to calprotectin in clinical practice. Fecal lactoferrin is the only possible exception, but a more extensive investigation is still required.

Retained particle surface area dose drives inflammation in rat lungs following acute, subacute, and subchronic inhalation of nanomaterials

Background An important aspect of nanomaterial (NM) risk assessment is establishing relationships between physicochemical properties and key events governing the toxicological pathway leading to adverse outcomes. The difficulty of NM grouping can be simplified if the most toxicologically relevant dose metric is used to assess the toxicological dose-response. Here, we thoroughly investigated the relationship between acute and chronic inflammation (based on polymorphonuclear neutrophil influx (% PMN) in lung bronchoalveolar lavage) and the retained surface area in the lung. Inhalation studies were performed in rats with three classes of NMs: titanium dioxides (TiO2) and carbon blacks (CB) as poorly soluble particles of low toxicity (PSLT), and multiwall carbon nanotubes (MWCNTs). We compared our results to published data from nearly 30 rigorously selected articles. Results This analysis combined data specially generated for this work on three benchmark materials - TiO2 P25, the CB Printex-90 and the MWCNT MWNT-7 - following subacute (4-week) inhalation with published data relating to acute (1-week) to subchronic (13-week) inhalation exposure to the classes of NMs considered. Short and long post-exposure recovery times (immediately after exposure up to more than 6 months) allowed us to examine both acute and chronic inflammation. A dose-response relationship across short-term and long-term studies was revealed linking pulmonary retained surface area dose (measured or estimated) and % PMN. This relationship takes the form of sigmoid curves, and is independent of the post-exposure time. Curve fitting equations depended on the class of NM considered, and sometimes on the duration of exposure. Based on retained surface area, long and thick MWCNTs (few hundred nm long with an aspect ratio greater than 25) had a higher inflammatory potency with 5 cm2/g lung sufficient to trigger an inflammatory response (at 6% PMN), whereas retained surfaces greater than 150 cm2/g lung were required for PSLT. Conclusions Retained surface area is a useful metric for hazard grouping purposes. This metric would apply to both micrometric and nanometric materials, and could obviate the need for direct measurement in the lung. Indeed, it could alternatively be estimated from dosimetry models using the aerosol parameters (rigorously determined following a well-defined aerosol characterization strategy).

Synovial fluid α defensin has comparable accuracy to synovial fluid white blood cell count and polymorphonuclear percentage for periprosthetic joint infection diagnosis

Aims The aim of this study was to determine the diagnostic accuracy of α defensin (AD) lateral flow assay (LFA) and enzyme-linked immunosorbent assay (ELISA) tests for periprosthetic joint infection (PJI) in comparison to conventional synovial white blood cell (WBC) count and polymorphonuclear neutrophil percentage (PMN%) analysis. Methods Patients undergoing joint aspiration for evaluation of pain after total knee arthroplasty (TKA) or total hip arthroplasty (THA) were considered for inclusion. Synovial fluids from 99 patients (25 THA and 74 TKA) were analyzed by WBC count and PMN% analysis, AD LFA, and AD ELISA. WBC and PMN% cutoffs of ≥ 1,700 cells/mm3 and ≥ 65% for TKA and ≥ 3,000 cells/mm3 and ≥ 80% for THA were used, respectively. A panel of three physicians, all with expertise in orthopaedic infections and who were blinded to the results of AD tests, independently reviewed patient data to diagnose subjects as with or without PJI. Consensus PJI classification was used as the reference standard to evaluate test performances. Results were compared using McNemar’s test and area under the receiver operating characteristic curve (AUC) analysis. Results Expert consensus classified 18 arthroplasies as having failed due to PJI and 81 due to aseptic failure. Using these classifications, the calculated sensitivity and specificity of AD LFA was 83.3% (95% confidence interval (CI) 58.6 to 96.4) and 93.8% (95% CI 86.2 to 98.0), respectively. Sensitivity and specificity of AD ELISA was 83.3% (95% CI 58.6 to 96.4) and 96.3% (95% CI 89.6 to 99.2), respectively. There was no statistically significant difference between sensitivity (p = 1.000) or specificity (p = 0.157) of the two AD assays. AUC for AD LFA was 0.891. In comparison, AUC for synovial WBC count, PMN%, and the combination of the two values was 0.821 (sensitivity p = 1.000, specificity p < 0.001), 0.886 (sensitivity p = 0.317, specificity p = 0.011), and 0.926 (sensitivity p = 0.317, specificity p = 0.317), respectively. Conclusion The diagnostic accuracy of synovial AD for PJI diagnosis is comparable and not statistically superior to that of synovial WBC count plus PMN% combined. Cite this article: Bone Joint J 2021;103-B(6):1119–1126.

Determining the Risk of Spontaneous Bacterial Peritonitisdue to increase use of Proton Pump Inhibitors among cirrhotic patients with ascites

Objectives: The current study aimed to determine the Spontaneous Bacterial Peritonitis (SBP) risk due to increased use of Proton Pump Inhibitors (PPIs) among cirrhotic patients with ascites. Methods: This retrospective case-control study was conducted at Chandka Medical College & Hospital, Larkana from March 2013 to February 2014, involving 215 cirrhotic patients with ascites. Paracentesis was performed to distinguish cirrhotic patients with SBP and Polymorphonuclear Neutrophil (PMN) count ≥ 250 neutrophils/mm3 (cases) and non-SBP with PMN count < 250 neutrophils/mm3 (controls). The demographic details, history of PPIs use before admission and duration of Chronic Liver Disease (CLD) were inquired and statistical analysis was carried through SPSS Version 23.0. Results: Increased pre-hospital PPI intake was observed among cirrhotic patients with SBP (69.8%) as compared to those without SBP (48.8%; p = 0.014). The mean duration of PPI use was 19.16 ± 4.772 days, and it was more significant among older cirrhotic patients (p < 0.05). Increased duration of CLD was observed among PPI users, i.e. 20.47 ± 6.305 months vs. 18.95 ± 5.527 months among non-PPI users (p < 0.05). Conclusions: Our results show that cirrhotic patients with ascites consuming PPIs are more likely to develop SBP as compared to non-PPI users. doi: https://doi.org/10.12669/pjms.37.4.3476 How to cite this:Shaikh BA, Shaikh ZA, Shah AH, Kumar A. Determining the Risk of Spontaneous Bacterial Peritonitis due to increase use of Proton Pump Inhibitors among cirrhotic patients with ascites. Pak J Med Sci. 2021;37(4):---------. doi: https://doi.org/10.12669/pjms.37.4.3476 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.