Polymorphonuclear Leukocyte Infiltration Research Articles

Evaluation of the Efficacy of Ozone Therapy on Liver Tissue in the Treatment of Sepsis in Rats with Cecal Perforation.

Background and Objectives: Sepsis and its related complications are associated with high morbidity and mortality, often leading to liver damage. Ozone, a molecule with anti-inflammatory and antioxidant properties, may offer protective effects. This study aimed to evaluate the therapeutic and protective impact of ozone on liver injury in a rat model of sepsis induced by cecal ligation and perforation (CLP). Material and Methods: A total of 36 rats were randomly divided into five groups: control (Group C), ozone (Group O), cecal ligation and perforation (Group CLP), ozone + cecal ligation and perforation (Group O+CLP), and cecal ligation and perforation + ozone (Group CLP+O). In the ozone groups, 4 mL of ozone (20 µ/mL) was injected intraperitoneally. Biochemical and histopathological parameters were evaluated in liver tissue samples obtained at the end of 24 h. Results: Polymorphonuclear leukocyte and monocyte infiltration and the total injury score were significantly reduced in the ozone-treated groups compared to the CLP group (p < 0.001). Tumor necrosis factor and interleukin 10 levels in the rat liver tissue were significantly reduced in the O+CLP and CLP+O groups compared to the CLP group, with the O+CLP group showing a more substantial decrease than the CLP+O group (p < 0.001). Thiobarbituric acid reactive substances and glutathione s-transferase levels were significantly lower in the ozone-treated groups compared to the CLP group (p < 0.001). Catalase activity was significantly elevated in the O+CLP group compared to the CLP group (p < 0.001). Serum aspartate transaminase, alanine transaminase, gamma-glutamyl transferase, and total bilirubin were significantly increased in the CLP group and decreased in the ozone-treated groups (p < 0.001, p < 0.001, p = 0.01, p < 0.001 respectively). Conclusions: Administering ozone to rats one hour before the CLP significantly mitigated liver damage, showing a more pronounced effect compared to administering ozone one hour after CLP. The results indicate that ozone could serve a protective function in managing sepsis-induced liver damage.

Blocking HXA3-mediated neutrophil elastase release during S. pneumoniae lung infection limits pulmonary epithelial barrier disruption and bacteremia.

Streptococcus pneumoniae (Sp), a leading cause of community-acquired pneumonia, can spread from the lung into the bloodstream to cause septicemia and meningitis, with a concomitant threefold increase in mortality. Limitations in vaccine efficacy and a rise in antimicrobial resistance have spurred searches for host-directed therapies that target pathogenic immune processes. Polymorphonuclear leukocytes (PMNs) are essential for infection control but can also promote tissue damage and pathogen spread. The major Sp virulence factor, pneumolysin, triggers acute inflammation by stimulating the 12-lipoxygenase (12-LOX) eicosanoid synthesis pathway in epithelial cells. This pathway is required for systemic spread in a mouse pneumonia model and produces a number of bioactive lipids, including hepoxilin A3 (HXA3), a hydroxy epoxide PMN chemoattractant that has been hypothesized to facilitate breach of mucosal barriers. To understand how 12-LOX-dependent inflammation promotes dissemination during Sp lung infection and dissemination, we utilized bronchial stem cell-derived air-liquid interface cultures that lack this enzyme to show that HXA3 methyl ester (HXA3-ME) is sufficient to promote basolateral-to-apical PMN transmigration, monolayer disruption, and concomitant Sp barrier breach. In contrast, PMN transmigration in response to the non-eicosanoid chemoattractant N-formyl-L-methionyl-L-leucyl-phenylalanine (fMLP) did not lead to epithelial disruption or bacterial translocation. Correspondingly, HXA3-ME but not fMLP increased the release of neutrophil elastase (NE) from Sp-infected PMNs. Pharmacologic blockade of NE secretion or activity diminished epithelial barrier disruption and bacteremia after pulmonary challenge of mice. Thus, HXA3 promotes barrier-disrupting PMN transmigration and NE release, pathological events that can be targeted to curtail systemic disease following pneumococcal pneumonia.IMPORTANCEStreptococcus pneumoniae (Sp), a leading cause of pneumonia, can spread from the lung into the bloodstream to cause systemic disease. Limitations in vaccine efficacy and a rise in antimicrobial resistance have spurred searches for host-directed therapies that limit pathologic host immune responses to Sp. Excessive polymorphonuclear leukocyte (PMN) infiltration into Sp-infected airways promotes systemic disease. Using stem cell-derived respiratory cultures that reflect bona fide lung epithelium, we identified eicosanoid hepoxilin A3 as a critical pulmonary PMN chemoattractant that is sufficient to drive PMN-mediated epithelial damage by inducing the release of neutrophil elastase. Inhibition of the release or activity of this protease in mice limited epithelial barrier disruption and bacterial dissemination, suggesting a new host-directed treatment for Sp lung infection.

Effects of Sevoflurane and Fullerenol C60 on the Heart and Lung in Lower-Extremity Ischemia-Reperfusion Injury in Streptozotocin-Induced Diabetes Mice.

Background and Objectives: Lower-extremity ischemia-reperfusion injury can induce distant organ ischemia, and patients with diabetes are particularly susceptible to ischemia-reperfusion injury. Sevoflurane, a widely used halogenated inhalation anesthetic, and fullerenol C60, a potent antioxidant, were investigated for their effects on heart and lung tissues in lower-extremity ischemia-reperfusion injury in streptozotocin (STZ)-induced diabetic mice. Materials and Methods: A total of 41 mice were divided into six groups: control (n = 6), diabetes-control (n = 7), diabetes-ischemia (n = 7), diabetes-ischemia-fullerenol C60 (n = 7), diabetes-ischemia-sevoflurane (n = 7), and diabetes-ischemia-fullerenol C60-sevoflurane (n = 7). Diabetes was induced in mice using a single intraperitoneal dose of 55 mg/kg STZ in all groups except for the control group. Mice in the control and diabetes-control groups underwent midline laparotomy and were sacrificed after 120 min. The DIR group underwent 120 min of lower-extremity ischemia followed by 120 min of reperfusion. In the DIR-F group, mice received 100 μg/kg fullerenol C60 intraperitoneally 30 min before IR. In the DIR-S group, sevoflurane and oxygen were administered during the IR procedure. In the DIR-FS group, fullerenol C60 and sevoflurane were administered. Biochemical and histological evaluations were performed on collected heart and lung tissues. Results: Histological examination of heart tissues showed significantly higher necrosis, polymorphonuclear leukocyte infiltration, edema, and total damage scores in the DIR group compared to controls. These effects were attenuated in fullerenol-treated groups. Lung tissue examination revealed more alveolar wall edema, hemorrhage, vascular congestion, polymorphonuclear leukocyte infiltration, and higher total damage scores in the DIR group compared to controls, with reduced injury parameters in the fullerenol-treated groups. Biochemical analyses indicated significantly higher total oxidative stress, oxidative stress index, and paraoxonase-1 levels in the DIR group compared to the control and diabetic groups. These levels were lower in the fullerenol-treated groups. Conclusions: Distant organ damage in the lung and heart tissues due to lower-extremity ischemia-reperfusion injury can be significantly reduced by fullerenol C60.

A Molecular and Histopathological Study on Bronchopneumonia in Cats

In this study, it was aimed to determine Bordetella bronchiseptica, Mycoplasma felis, Staphylococcus aureus and Chlamydia felis, which cause bronchopneumonia in cats, by Real-time PCR and to compare the pathological findings of the identified agents. The material of the study was constituted of paraffin blocks belonging to the lungs, of which 21 bronchopneumonia were detected in microscopic examination (with Hematoxylin Eosin (HE)) from a total of 78 cats samples brought to Selcuk University, Faculty of Veterinary Medicine, Department of Pathology for pathological diagnosis. Histopathologically, polymorphonuclear leukocytes (PMNL) and mononuclear cell infiltration (MCI) in the bronchi and bronchiolar lumens, desquamation in the bronchi and bronchiolar epithelium, PMNL infiltration with oedama in alveolar lumens and desquamated alveolar epithelium, PMNL infiltration in the interstitium, and peribronchi and peribronchiolar MCI, and pleuritis were detected. Real-time PCR analysis revealed Bordetella bronchiseptica in 3 (14.29%) cases, Mycoplasma felis in 3 (14.29%), Staphylococcus aureus in 5 (23.8%), and Chlamydia felis in 5 (23.8%). Morever, Mycoplasma felis and Staphylococcus aureus infection was detected in 1 case, and Staphylococcus aureus and Chlamydia felis mixed infection was observed in 1 case. Our results show that relevant agents can frequently be isolated in cases of feline bronchopneumonia.

Effect of Curcuma longa on GPER-1 and Oxidative/Nitrosative Stress Biomarkers in Cardiac Ischemia-reperfusion Injury

Background: The death rate caused by cardiovascular diseases is increasing all over the world. To investigate the cardioprotective effects of Curcuma longa on cardiac ischemia-reperfusion (I/R) injury, which was further examined through G protein-coupled estrogen receptor 1 (GPER-1) and oxidative/nitrosative stress biomarkers. Methods: Twenty-four male Wistar-albino rats were divided into three groups (I/R group; n=8, sham group; n=8, treatment (Curcuma longa) group; n=8). 1 ml Curcuma longa 50 mg/kg was administered intraperitoneally in the treatment group. While catalase, superoxide dismutase and malondialdehyde values were measured by spectrophotometric method, the measurement of GPER-1 was performed by ELISA method. In the histopathological examination, the groups were evaluated in terms of hemorrhage, myocardial edema, myocytolysis and polymorphonuclear leukocyte (PMNL) infiltration. Result: Statistically significant differences were observed between the treatment and I/R groups in terms of oxidative stress (malondialdehyde) level and antioxidant enzyme activities (catalase, superoxide dismutase) (p less than 0.05). The GPER-1 level was higher in the treatment group compared to the I/R group (p less than 0.05). Histopathologically, PMNL infiltration, hemorrhage, edema and myocytolysis were observed in the I/R and sham groups, whereas only hemorrhage was detected in the treatment group. This study showed that treatment with Curcuma longa inhibited lipid peroxidation by restoring existing antioxidant enzymes against cardiac ischemia-reperfusion injuries.

Propionyl-l-carnitine mitigates ischemia-reperfusion injury in rat epigastric island flaps

BackgroundIschemia-reperfusion injury presents a substantial concern in various medical scenarios, notably in reconstructive surgery involving tissue flaps. Despite reports on the protective benefits of Propionyl-l-carnitine against ischemia-reperfusion injury, a thorough assessment of its efficacy in epigastric island flap models is currently lacking. MethodsSixteen male Sprague-Dawley rats underwent epigastric island flap surgery and were divided into two groups: a Propionyl-l-carnitine group that received intraperitoneal Propionyl-l-carnitine prior to ischemia induction and a sham group that received saline treatment. A comprehensive evaluation was performed including macroscopic, biochemical and histological assessments encompassing measurements of flap survival areas, lipid peroxidation (malondialdehyde), glutathione, myeloperoxidase, nitric oxide and peripheral neutrophil counts. ResultsThe Propionyl-l-carnitine group demonstrated significantly increased flap survival areas when compared to the sham group. Administration of Propionyl-l-carnitine led to reduced malondialdehyde levels and elevated glutathione levels indicating a reduction in oxidative stress. Furthermore, the Propionyl-l-carnitine group exhibited lower myeloperoxidase levels, higher nitric oxide levels and reduced peripheral neutrophil counts, suggesting a decrease in the inflammatory response. Histopathological analysis revealed decreased levels of inflammation, necrosis, polymorphonuclear leukocyte infiltration and edema in the Propionyl-l-carnitine group. Additionally, vascularity was enhanced in the Propionyl-l-carnitine group. ConclusionThis study provides compelling evidence that Propionyl-l-carnitine administration effectively mitigates the deleterious effects of ischemia-reperfusion injury in epigastric island flaps. This is substantiated by the improved flap survival, diminished oxidative stress and inflammation, as well as the enhanced vascularity observed. Propionyl-l-carnitine emerges as a promising therapeutic intervention to enhance tissue flap survival in reconstructive surgery, warranting further exploration through larger-scale investigations.

Effectivity of African leaf extract (Vernonia amygdalina) in wound healing: a systematic review

Background: The use of natural products, particularly those derived from plants, for nutritional and medical purposes to enhance human performance, including wound healing, has gained more attention in recent years. Prior studies showed the health benefits of African leaf (Vernonia amygdalina) as anti-parasitic, anti-malarial, anti-helminthic, antiviral, anticancer, anticoagulant, antithrombotic, analgesic, antipyretic effects, including the wound healing process. However, there are still mixed results regarding its effectiveness. This systematic review aims to elaborate on the effectiveness of African leaf extract in wound healing. Method: A systematic review using the PRISMA diagram was conducted based on literature searching on three online databases (Google Scholar, Pubmed, and Cochrane Library). An eligibility criteria was applied to the literature search, and the Jadad scoring system was used to assess the study's methodological quality. Result: We included ten clinical trial studies regarding the role and effectiveness of African leaves in wound healing. Studies come from Indonesia, Malaysia, and Nigeria. The animal models used were Wistar albino rats, rabbits, and mice. The intervention used was African leaf extract as a monotherapy or a combination of African leaf extract with other natural agents. The types used were burn wounds, incision or excision wounds, and diabetic wounds. The outcome measured was the wound healing process marked by re-epithelization, wound healing and closure time, and histopathological evaluation to assess collagen formation, polymorphonuclear leukocyte infiltration, and fibroblast recruitment. Most of the included studies showed positive results of African leaf extract or combination with other agents in wound healing. Conclusion: African leaf (Vernonia amygdalina) showed effectiveness in the wound healing process in terms of facilitating re-epithelization, fastened wound healing time, and significant wound reduction size compared with the control group.

Comparison of the protective effects of vanillic and rosmarinic acid on cardiac tissue: Lower limb ischemia-reperfusion model in rats.

Ischemia/reperfusion injury is one of the most challenging postoperative situations in vascular surgery, both in elective procedures with prolonged clamping time and in delayed emergency cases with vascular occlusion. The inflammatory response that develops during ischemia and the oxygen-free radicals that proliferate during reperfusion have detrimental effects on the brain, heart, and kidneys. In this study, we aimed to compare the effects of vanillic and rosmarinic acid in preventing ischemia/reperfusion injury in a lower limb ischemia-reperfusion model in rats. Thirty-two female Sprague-Dawley rats weighing 185-240 g were randomly divided into four groups of eight animals each. Group 1 was designated as the control, Group 2 as ischemia/reperfusion (I/R), Group 3 as ischemia/reperfusion + vanillic acid (I/R + VA), and Group 4 as ischemia/reperfusion + rosmarinic acid (I/R + RA). In all groups except the control, the infrarenal abdominal aorta was clamped, and 60 minutes of ischemia followed by 120 minutes of reperfusion was performed. Vanillic acid was administered intra-abdominally 15 minutes before the start of reperfusion in Group 3, and rosmarinic acid in Group 4. At the end of the reperfusion phase, blood samples and hearts were collected, and the rats were euthanized. Histopathologically, myofibrillar edema, myocytolysis, focal hemorrhages, and infiltration of polymorphonuclear leukocytes (PMNL) in cardiac tissue were examined. Total antioxidant capacity (TAC), total oxidative status (TOS), oxidative stress index (OSI), 8-OH-deoxyguanosine, lactonase, and arylesterase activity were measured in blood samples. Myofibrillar edema was most pronounced in the I/R group and less pronounced in the I/R + VA and I/R + RA groups (p=0.005 and p=0.066, respectively). There was no difference between the ischemia/reperfusion groups regarding myocytolysis, focal hemorrhage, and PMNL infiltration (p>0.99). Among all groups, TOS and OSI were lowest in the control group, while TAC was highest. TAC was similar in the I/R + VA and I/R + RA groups but was significantly higher in these two groups than in the I/R group. The lactonase activity in the I/R + VA group was similar to that in the control group but was significantly higher compared to the I/R and I/R + RA groups. Our study shows that vanillic and rosmarinic acids reduce myofibrillar edema in the heart after lower limb ischemia and increase TAC. However, vanillic acid increases the activity of lactonase, an enzyme known for its antioxidant effect, more than rosmarinic acid.

Effect of humic acid on oxidative stress and neuroprotection in traumatic spinal cord injury: an experimental study.

Traumatic spinal cord injury (TSCI) is an important health problem, especially in developing countries with additional socioeconomic loss. Humic acid (HA) usually has antioxidant, antiinflammatory, blood circulating, and antiviral effects. Hence, it was aimed herein to show the effect of HA on neuroprotection in a TSCI model. A TSCI model was used, in which 24 Wistar albino rats were divided into 4 groups: control group: subjected to only laminectomy; sham group: subjected to laminectomy + TSCI; HA 5 mg/kg group: subjected to laminectomy + TSCI + intraperitoneal (IP) injection of 5 mg/kg of HA; and HA 10 mg/kg group: subjected to laminectomy + TSCI + IP injection of 10 mg/kg of HA. Intracardiac blood samples were obtained preoperatively (preop), and at 1 and 24 h postoperatively (postop). The total antioxidant status (TAS), total oxidant status (TOS) and oxidative stress index (OSI) levels were evaluated in the serum. The motor functions were evaluated using the Modified Tarlov Score at 24 h postop. There were no significant changes in the TAS values between the sham and HA 5 mg/kg and HA 10 mg/kg groups (p = 0.77/0.21). However there was a significant decrease in the TOS values at 24 h postop when comparing the sham and HA 5 mg/kg groups (p = 0.02). The pathological evaluation showed a significant decrease in the severity of edema, hemorrhage, polymorphonuclear leucocyte (PNL) infiltration, and mononuclear leucocyte (MNL)/macrophage/microglia infiltration when compared with the control group (p < 0.05). There was a significant recovery at the paraplegia level when the HA 5 mg/kg and HA 10 mg/kg groups were compared with the control group (p < 0.001). The effects of HA in the early stages of TSCI on oxidative stress, histopathological changes, and neurological improvement were investigated herein. It is thought to be a potential therapeutic agent in acute TSCI but needs to be further evaluated to determine the extent of its effect on other neuroprotective pathways in larger series.

Bronkopnömonili Köpeklerde Mycoplasma spp., Streptococcus spp., Bordetella bronchiseptica, Klebsiella spp. ’nin Real-time PCR ve Patolojik Yöntemler ile Teşhisi

There are many bacterial factors that cause respiratory diseases in dogs. Bacterial bronchopneumonia is a lung disease caused by bacteria in the lower respiratory tract and lung parenchyma. In this study, it was aimed to determine Mycoplasma spp., Streptococcus spp., Bordetella bronchiseptica, Klebsiella spp., in dogs with bronchopneumonia by Real-time PCR and to compare the histopathological findings determined according to the agents. The material of the study consisted of lung paraffin blocks of 37 dogs that died due to different reasons and were found to have bronchopneumonia in microscopic examination. Microscopically, edema in the alveolar lumens, shedding of the alveolar epithelium, shedding of the bronch and bronchial epithelium and bronchiectasis, mononuclear cell infiltration (MCI) and Polymorphonuclear leukocytes (PMNL) infiltration in the bronchial lumens, peribronchiolar MCI infiltration, Bacterial clusters localized to the bronch epithelium, PMNL infiltration in the interstitium, multifocal necrosis areas, bleeding and pleuritis were observed. Real-time PCR analysis revealed Bordetella bronchiseptica in 18 (48.64%) cases, Mycoplasma spp. in 9 (24.32%) cases, Streptococcus spp. in 10 (27.02%) cases, and Klebsiella spp. in 2 (5.4%) cases. In conclusion, this study showed that the causative agent can be determined in bacterial bronchopneumonias of dogs with Real-time PCR even in tissues without culture opportunity. In addition, this study indicates that polymicrobial lower respiratory tract infections can also be seen in dogs and reveals that more than one bacterial species should be investigated for diagnosis.

Agomelatine on cisplatin-induced nephrotoxicity via oxidative stress and apoptosis.

Drug-induced nephrotoxicity is the greatest deterrent to the use of cisplatin, which is a frequently used chemotherapeutic with proven effectiveness in cancer therapy. Agomelatine, which is used in the treatment of sleep disorders and depression, has gained attention in recent years with its antioxidative and anti-inflammatory effects. In this study, the effects of the synthetic melatonin agonist agomelatine on nephrotoxicity were investigated in a rat model of cisplatin-induced nephrotoxicity using biochemical, histological, and immunohistochemical methods. Thirty-two male rats were divided into 4 groups: 1. control group, 2. agomelatine group, 3. cisplatin group, 4. cisplatin + agomelatine group. In the cisplatin group, there were widespread atypical glomerular structures and vacuolization in tubular epithelial cells, necrotic tubules, deterioration of brush border structure in proximal tubules, and fibrotic areas characterized by diffuse polymorphonuclear leukocyte (PNL) and extensive collagen deposition in the interstitial spaces. However, in the cisplatin + agomelatine group, we observed a reduction in glomeruli of atypical structure and necrotic tubules, in PNL infiltration in interstitial spaces, and fibrotic areas compared to the cisplatin group. The cisplatin + agomelatine group showed lower malondialdehyde (MDA) serum creatinine, serum urea levels, and higher glutathione (GSH) levels compared to the cisplatin group. Immunohistochemical analyses revealed that the elevated NF-kβ/p65, 8-OHdG, and cleaved caspase-3 positivity in the cisplatin group had significantly decreased in the cisplatin + agomelatine group. In conclusion, agomelatine showed a nephroprotective effect against cisplatin-induced nephrotoxicity.

Strangles in a stallion in southeast Nigeria: clinical and pathological report

ABSTRACT. Streptococcus equi subspecies equi is the causative agent of strangles, a highly contagious mucopurulent upper respiratory tract infection of horses. This report describes the clinicopathologic findings of a fatal case of pulmonary form of strangles in a two-and-half-year-old stallion presented at a Veterinary Teaching Hospital in Southeast Nigeria. Treatment was attempted in quarantine using high doses of procaine penicillin; but it was unsuccessful. The clinical signs observed in the affected horse include a left unilateral supra-orbital swelling containing purulent matter, which progressed to a chronic intractable exudative pus. Anaemia with leukocytosis (due to neutrophilia) were observed in the haematologic parameters. Gross and histologic lesions were multifocally extensive abscessing necrotic lymphadenitis and bronchopneumonia with inflammatory oedema and infiltrate of polymorphonuclear leukocytes.

The Effect of Desloratadine Upon Ischemia-reperfusion-induced Oxidative and Inflammatory Gastric Injury in Rats

Background: Ischemia-reperfusion (I/R) injury is a pathological process mediated by oxidative stress and inflammation that begins with oxygen deprivation. This study, it was aimed to investigate the protective effect of desloratadine on gastric I/R damage in rats. Methods: Eighteen rats were randomly divided into 3 groups gastric I/R (GIR), gastric I/R + 5 mg/kg oral desloratadine (GIRD) and gastric sham operation (GSG). One hour later desloratadine administration, clips were placed in the celiac arteries of the rats in the GIRD and GIR groups. Ischemia was created by keeping the clips closed for one hour. Afterward, the clips were removed and reperfusion was achieved for three hours. The sham operation was performed on the GSG group. Then, all animals were euthanized with 120 mg/kg ketamine. Malondialdehyde (MDA), total glutathione (tGSH), superoxide dismutase (SOD) catalase (CAT) and interleukin-6 (IL-6) levels were determined in the excised gastric tissues. All tissues were also analyzed histopathologically. Result: When the GIR group was compared with the GSG, an increase in MDA and IL-6 levels and a decrease in tGSH, SOD and CAT levels were observed (p less than 0.001). In addition, severe mucosal degeneration, congestion, polymorphonuclear leukocyte infiltration and mucosal edema were detected in the GIR group. Desloratine significantly inhibited both biochemical and histopathological changes (p less than 0.05).

Mitigating neutrophil trafficking and cardiotoxicity with DS-IkL in a microphysiological system of a cytokine storm.

A feature of severe COVID-19 is the onset of an acute and intense systemic inflammatory response referred to as the "cytokine storm". The cytokine storm is characterized by high serum levels of inflammatory cytokines and the subsequent transport of inflammatory cells to damaging levels in vital organs (e.g., myocarditis). Immune trafficking and its effect on underlying tissues (e.g., myocardium) are challenging to observe at a high spatial and temporal resolution in mouse models. In this study, we created a vascularized organ-on-a-chip system to mimic cytokine storm-like conditions and tested the effectiveness of a novel multivalent selectin-targeting carbohydrate conjugate (composed of DS - dermatan sulfate and IkL - a selectin-binding peptide, termed DS-IkL) in blocking infiltration of polymorphonuclear leukocytes (PMN). Our data shows that cytokine storm-like conditions induce endothelial cells to produce additional inflammatory cytokines and facilitate infiltration of PMNs into tissue. Treatment of tissues with DS-IkL (60 μM) reduced PMN accumulation in the tissue by >50%. We then created cytokine storm-like conditions in a vascularized cardiac tissue-chip and found that PMN infiltration increases the spontaneous beating rate of the cardiac tissue, and this effect is eliminated by treatment with DS-IkL (60 μM). In summary, we demonstrate the utility of an organ-on-a-chip platform to mimic COVID-19 related cytokine storm and that blocking leukocyte infiltration with DS-IkL could be a viable strategy to mitigate associated cardiac complications.

Maternal Ethanol Exposure Impairs the Kidney of Offspring by Alteration of Podocyte Proteins Genes Expression and Fibrosis

Background: This study examined the effect of prenatal and early postnatal ethanol exposure on the structural, functional, and molecular alterations of rat’s offspring kidney on postnatal days 21 and 90.Methods: Pregnant rats on gestation day 7 were divided into the two groups, namely control and ethanol groups. Rats in the ethanol group received ethanol (4.5 g/kg B.W) from gestation day 7 throughout lactation. Nephrin, podocin, vascular endothelial growth factor receptors (VEGFRs) 1 and 2 gene expression were measured by RT-PCR technique. The MMP2 and MMP9 levels in the kidney tissue and plasma cystatin C level were measured by ELISA method.Results: The results revealed a significant alteration in mRNA expression of nephrin, podocin, and VEGFR, as well as MMPs amounts in the kidneys of the offspring. Cystatin C level, the ratio of cystatin C/serum creatinine, serum creatinine, and urine urea showed a significant increase, but urine creatinine and GFR showed a significant decrease in the offsprings of the ethanol group compared to the control group. Histopathological changes such as fibrosis, kidney cells proliferation, leukocytes infiltration, and vacuolization have also seen in the kidney of the offsprings after 21 and 90 days from birth.Conclusion: Taken together, these results provide evidence that pre and early postnatal ethanol exposure renal toxicity is in part associated with alteration of nephrin, podocin, and VEGFRs genes expression, as well as MMPs amount changes. Furthermore, it was found that these molecular alterations were triggered by inflammatory reactions manifested by fibrosis, proliferation, and polymorphonuclear(PMN) leukocytes infiltration.

Therapeutic Targeting of NF-κB in Acute Lung Injury: A Double-Edged Sword.

Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) is a devastating disease that can be caused by a variety of conditions including pneumonia, sepsis, trauma, and most recently, COVID-19. Although our understanding of the mechanisms of ALI/ARDS pathogenesis and resolution has considerably increased in recent years, the mortality rate remains unacceptably high (~40%), primarily due to the lack of effective therapies for ALI/ARDS. Dysregulated inflammation, as characterized by massive infiltration of polymorphonuclear leukocytes (PMNs) into the airspace and the associated damage of the capillary-alveolar barrier leading to pulmonary edema and hypoxemia, is a major hallmark of ALI/ARDS. Endothelial cells (ECs), the inner lining of blood vessels, are important cellular orchestrators of PMN infiltration in the lung. Nuclear factor-kappa B (NF-κB) plays an essential role in rendering the endothelium permissive for PMN adhesion and transmigration to reach the inflammatory site. Thus, targeting NF-κB in the endothelium provides an attractive approach to mitigate PMN-mediated vascular injury, not only in ALI/ARDS, but in other inflammatory diseases as well in which EC dysfunction is a major pathogenic mechanism. This review discusses the role and regulation of NF-κB in the context of EC inflammation and evaluates the potential and problems of targeting it as a therapy for ALI/ARDS.

The role of polymorphonuclear leukocytes in distant organ (lung) oxidative damage of liver ischemia/reperfusion and the protective effect of rutin.

Ischemia/reperfusion (I/R) can cause damage to distant organs. Rutin is known to have antioxidant and anti-inflammatory properties, and inhibits cytokine and polymorphonuclear leukocyte (PMNL) infiltration. It may prevent the development of reperfusion injury. This study aimed to examine the role of PMNLs in distant organ (lung) injury after a liver I/R procedure, and to evaluate the protective effects of rutin in rats using biochemical and immunohistochemical methods. In this study, 18 Wistar albino male rats (255-275 g) were used. Experimental animals were divided into 3 groups: a liver I/R (LIR) group, a 50 mg/kg rutin+liver I/R (RLIR) group and a sham operation (SG) control group. Experimental results obtained from the RLIR group were compared with the LIR and SG groups. Blood malondialdehyde (MDA) levels in the RLIR and SG groups were significantly lower compared to the LIR group (p < 0.001). Blood myeloperoxidase (MPO) activity in the RLIR and SG groups was significantly lower compared to the LIR group (p < 0.001). Total glutathione (tGSH) levels in the RLIR and SG groups were significantly higher compared to the LIR group (p < 0.001). Rutin can be used to prevent distant organ (lung) damage due to liver I/R. However, more extensive studies are needed on this issue.

Platelet Endothelial Cell Adhesion Molecule (PECAM/CD31) Blockade Modulates Neutrophil Recruitment Patterns and Reduces Infarct Size in Experimental Ischemic Stroke

The infiltration of polymorphonuclear leukocytes (PMNs) in ischemia-reperfusion injury (I/RI) has been implicated as a critical component of inflammatory damage following ischemic stroke. However, successful blockade of PMN transendothelial migration (TEM) in preclinical studies has not translated to meaningful clinical outcomes. To investigate this further, leukocyte infiltration patterns were quantified, and these patterns were modulated by blocking platelet endothelial cell adhesion molecule-1 (PECAM), a key regulator of TEM. LysM-eGFP mice and microscopy were used to visualize all myeloid leukocyte recruitment following ischemia/reperfusion. Visual examination showed heterogeneous leukocyte distribution across the infarct at both 24 and 72 hours after I/RI. A semiautomated process was designed to precisely map PMN position across brain sections. Treatment with PECAM function-blocking antibodies did not significantly affect total leukocyte recruitment but did alter their distribution, with more observed at the cortex at both early and later time points (24 hours: 89% PECAM blocked vs. 72% control; 72 hours: 69% PECAM blocked vs. 51% control). This correlated with a decrease in infarct volume. These findings suggest that TEM, in the setting of I/RI in the cerebrovasculature, occurs primarily at the cortical surface. The reduction of stroke size with PECAM blockade suggests that infiltrating PMNs may exacerbate I/RI and indicate the potential therapeutic benefit of regulating the timing and pattern of leukocyte infiltration after stroke.

Resolution of Inflammation after Skeletal Muscle Ischemia-Reperfusion Injury: A Focus on the Lipid Mediators Lipoxins, Resolvins, Protectins and Maresins.

Skeletal muscle ischemia reperfusion is very frequent in humans and results not only in muscle destruction but also in multi-organ failure and death via systemic effects related to inflammation and oxidative stress. In addition to overabundance of pro-inflammatory stimuli, excessive and uncontrolled inflammation can also result from defects in resolution signaling. Importantly, the resolution of inflammation is an active process also based on specific lipid mediators including lipoxins, resolvins and maresins that orchestrate the potential return to tissue homeostasis. Thus, lipid mediators have received growing attention since they dampen deleterious effects related to ischemia–reperfusion. For instance, the treatment of skeletal muscles with resolvins prior to ischemia decreases polymorphonuclear leukocyte (PMN) infiltration. Additionally, remote alterations in lungs or kidneys are reduced when enhancing lipid mediators’ functions. Accordingly, lipoxins prevented oxidative-stress-mediated tissue injuries, macrophage polarization was modified and in mice lacking DRV2 receptors, ischemia/reperfusion resulted in excessive leukocyte accumulation. In this review, we first aimed to describe the inflammatory response during ischemia and reperfusion in skeletal muscle and then discuss recent discoveries in resolution pathways. We focused on the role of specialized pro-resolving mediators (SPMs) derived from polyunsaturated fatty acids (PUFAs) and their potential therapeutic applications.

Comparison of CD3e Antibody and CD3e-sZAP Immunotoxin Treatment in Mice Identifies sZAP as the Main Driver of Vascular Leakage.

Anti-CD3-epsilon (CD3e) monoclonal antibodies (mAbs) and CD3e immunotoxins (ITs) are promising targeted therapy options for various T-cell disorders. Despite significant advances in mAb and IT engineering, vascular leakage syndrome (VLS) remains a major dose-limiting toxicity for ITs and has been poorly characterized for recent “engineered” mAbs. This study undertakes a direct comparison of non-mitogenic CD3e-mAb (145-2C11 with Fc-silentTM murine IgG1: S-CD3e-mAb) and a new murine-version CD3e-IT (saporin–streptavidin (sZAP) conjugated with S-CD3e-mAb: S-CD3e-IT) and identifies their distinct toxicity profiles in mice. As expected, the two agents showed different modes of action on T cells, with S-CD3e-mAb inducing nearly complete modulation of CD3e on the cell surface, while S-CD3e-IT depleted the cells. S-CD3e-IT significantly increased the infiltration of polymorphonuclear leukocytes (PMNs) into the tissue parenchyma of the spleen and lungs, a sign of increased vascular permeability. By contrast, S-CD3e-mAbs-treated mice showed no notable signs of vascular leakage. Treatment with control ITs (sZAP conjugated with Fc-silent isotype antibodies) induced significant vascular leakage without causing T-cell deaths. These results demonstrate that the toxin portion of S-CD3e-IT, not the CD3e-binding portion (S-CD3e-mAb), is the main driver of vascular leakage, thus clarifying the molecular target for improving safety profiles in CD3e-IT therapy.