Granulocyte Research Articles

Case report: diagnosis of neurobrucellosis in a non-endemic area child using metagenomic next-generation sequencing.

Brucellosis with neurological symptoms at onset is rare in children and is frequently misdiagnosed or overlooked due to nonspecific clinical presentations, particularly in non-endemic areas. We report a case of neurobrucellosis in a child from a non-pastoral area, diagnosed via metagenomic next-generation sequencing (mNGS). The patient presented with headache and altered consciousness, accompanied by fever, projectile vomiting, seizures, and urinary incontinence. Physical examination indicated possible nuchal rigidity. Cerebrospinal fluid (CSF) analysis showed colorless and clear fluid, with a white blood cell count of 259 × 10⁶/L, 4.6% polymorphonuclear cells, positive protein qualitative test, protein level of 0.78g/L, and glucose level of 1.66 mmol/L. Initial diagnosis suggested central nervous system infection, and empirical treatment led to improvement in consciousness. However, after a few days of stable body temperature, the patient experienced recurrent fever. Ultimately, mNGS of CSF identified Brucella melitensis, confirming neurobrucellosis. Following treatment with ceftriaxone, doxycycline, and rifampin, the patient's clinical symptoms improved significantly, and follow-up CSF analysis showed normalization of cell counts. This case highlights the early diagnostic utility of mNGS in CSF for neurobrucellosis and its role in differential diagnosis.

Neuroendoscopy surgery for ventriculitis and hydrocephalus

Neuroendoscopy surgery is a minimally invasive surgical procedure that uses an endoscope to access and treat conditions within the brain. We are describing the case of a 2-month-old patient who was born extremely premature and developed complications from prematurity-related intraventricular hemorrhage. The mother had a preterm delivery due to chorioamnionitis, and the child developed neonatal sepsis and ventriculitis after birth. The child was treated conservatively from birth, without an effective response to treatment. A brain MRI showed tetraventricular hydrocephalus and several intraventricular collections. Cerebrospinal fluid analysis revealed increased cellularity with a predominance of polymorphonuclear cells, protein elevation, and glucose consumption. An exhaustive ventricular lavage, drainage of the intraventricular collections, septostomy, and third ventriculostomy were performed. The patient showed clinical and infectious improvement.

Efficacy of Plant Extract-Based Solutions Compared to Chlorhexidine and Miconazole Shampoo for the Treatment of Superficial Pyoderma in Dogs.

Canine superficial pyoderma (CSP) is common. Antibiotic and potential antiseptic resistance is a rising concern; natural topicals may provide valuable alternative options. Compare natural topicals containing essential oils, plant-extracted essential fatty acids and N-acetylcysteine to a medicated shampoo with chlorhexidine, miconazole and micronized silver in treating CSP. Thirty dogs diagnosed with CSP, divided into three groups as outlined below. Group 1 bathed using a shampoo (PYOclean Shampoo) twice weekly and application of a rinse-free mousse (PYOclean Mousse) once daily, Group 2 bathed using the same shampoo as above twice weekly and application of a spray (PYOclean Spray) once daily, and Group 3 bathed using a medicated shampoo (Biohex Shampoo) twice weekly. Cytology, lesion and pruritus visual analogue (PVAS) scores were evaluated on Days (D) 0, 7 and 15. All three groups showed decreased polymorphonuclear cells (PMN) on D15. The reduction was significant, and more significant in Groups 1 and 2 (90.48% and 98.93%) compared to Group 3 (85.96%); extracellular cocci (cocci EC) significantly decreased by 92.8% and 93.38% in Groups 1 and 2, whereas decreased by 81% in Group 3 on D15. The number of yeasts on D15 decreased by 90% in Group 3 and 82% and 92%, respectively, in Groups 1 and 2. Lesion scores decreased on D15 by 90.61% and 87.95% in Groups 1 and 2 and by 62.94% in Group 3. PVAS decreased on D15 by 95.78% and 96.75%, respectively, in Groups 1 and 2 and by 69.88% in Group 3. Efficacy of three natural topicals in treating dogs with superficial pyoderma without adding any systemic antimicrobial drugs was demonstrated. Such solutions can be complementary to medications such as chlorhexidine and miconazole in addressing CSP.

Gastric lesions associated with the infection of Anisakidae nematodes in a dwarf sperm whale Kogia sima (Owen, 1866) stranded in the north coast of Brazil

The present study aimed to describe gastric lesions associated with parasitism by different nematodes of the family Anisakidae in a stranded specimen of dwarf sperm whale (Kogia sima) on the northeast coast of the State of Pará, northern Brazil. Specimens of helminths and samples of stomach tissue were collected from a dwarf sperm whale, stranded on Humaitá beach, State of Pará, Brazil. Stomach showed areas of erosion and ulcers, with the mucosa covered by fibrinonecrotic material containing bacteria and inflammatory infiltrate predominantly comprising polymorphonuclear cells. Granulomas were also found in the submucosa, characterized by central areas of necrosis and hemorrhage, and cross sections of nematodes were observed. Fourth-stage larvae of the genus Pseudoterranova, two morphotypes of fourth-stage larvae of the genus Anisakis and adult specimens of Skrjabinisakis paggiae were morphologically identified. Molecular and phylogenetic analyzes confirmed the identity of the partial sequences of the cox2 mtDNA gene for adult specimens of S. paggiae. This study contributes to our understanding of the distribution of different of anisakids in K. sima and about the gastric lesions associated with these nematodes, in addition to expanding the knowledge about the occurrence of this aquatic mammal recorded for the first time in the northern region of Brazil.

Unveiling the Antibacterial Efficacy of a Benzonitrile Small Molecule, IITR00210, in Shigella Infection.

The escalating prevalence of bacterial infections and the rapid emergence of multidrug-resistant Gram-negative bacterial pathogens highlight an urgent demand for effective antibacterial agents. In this study, we report our findings on IITR00210, a small molecule belonging to the nitrile class. The small molecule demonstrates broad-spectrum activity against bacterial pathogens, specifically against enteric pathogens, and exhibits antibiofilm activity. IITR00210 displays potent bactericidal activity against enteropathogens, resulting in a reduction of bacterial counts greater than 3 Log10 CFU in time-kill kinetic assays. Mechanistic investigations revealed that IITR00210 induces bacterial cell envelope stress, leading to the alteration of the overall proton motive force (PMF). The disruption of PMF causes intracellular ATP dissipation and ultimately promotes cell death. The cell envelope stress generated in the presence of IITR00210 leads to a translational aberration. Importantly, IITR00210 exhibits a safe profile in in vitro and in vivo settings. The small molecule further showed potent intracellular antibacterial activity in polymorphonuclear cells infected with enteric pathogens and antiadhesion activity in mammalian cell lines. IITR00210 proves to be a promising therapeutic candidate, displaying a lack of stable resistance development, and it exhibited efficacy in the treatment of bacterial infections in a shigellosis murine model.

Neutrophil prime unique transcriptional responses in intestinal organoids during infection with nontyphoidal Salmonella enterica serovars.

Nontyphoidal strains of Salmonella enterica are a major cause of foodborne illnesses, and infection with these bacteria results in inflammatory gastroenteritis. Polymorphonuclear leukocytes (PMNs), also known as neutrophils, are a dominant immune cell type found at the site of infection in Salmonella-infected individuals, but how they regulate infection outcome is not well understood. Here, we used a co-culture model of primary human PMNs and human intestinal organoids to probe the role of PMNs during infection with two of the most prevalent Salmonella serovars: Salmonella enterica serovar Enteritidis and Typhimurium. Using a transcriptomics approach, we identified a dominant role for PMNs in mounting differential immune responses including production of pro-inflammatory cytokines, chemokines, and antimicrobial peptides. We also identified specific gene sets that were induced by PMNs in response to Enteritidis or Typhimurium infection. By comparing host responses to these serovars, we uncovered differential regulation of host metabolic pathways particularly induction of cholesterol biosynthetic pathways during Typhimurium infection and suppression of RNA metabolism during Enteritidis infection. Together, these findings provide insight into the role of human PMNs in modulating different host responses to pathogens that cause similar disease in humans.IMPORTANCENontyphoidal serovars of Salmonella enterica are known to induce robust recruitment of polymorphonuclear leukocytes (PMNs) in the gut during early stages of infection, but the specific role of PMNs in regulating infection outcome of different serovars is poorly understood. Due to differences in human infection progression compared to small animal models, characterizing the role of PMNs during infection has been challenging. Here, we used a co-culture model of human intestinal organoids with human primary PMNs to study the role of PMNs during infection of human intestinal epithelium. Using a transcriptomics approach, we define PMN-dependent reprogramming of the host response to Salmonella, establishing a clear role in amplifying pro-inflammatory gene expression. Additionally, the host response driven by PMNs differed between two similar nontyphoidal Salmonella serovars. These findings highlight the importance of building more physiological infection models to replicate human infection conditions to study host responses specific to individual pathogens.

Jordans’ anomaly in Chanarin-Dorfman syndrome

Abstract Objectives Chanarin-Dorfman syndrome is a rare disease inherited in an autosomal recessive pattern whose prevalence does not exceed 130 cases worldwide. Case presentation A 4-year-old patient with generalized erythematous-desquamative ichthyosiform syndrome since birth. The main laboratory finding was persistent hypertransaminasemia. Supplementary studies included peripheral blood smear (PBS), which revealed the presence of multiple cytoplasmatic vacuoles in polymorphonuclear leukocytes (PMN) and platelets. Ichthyosiform lesions concomitant to the presence of lipid vacuoles in peripheral blood PMNs are signs of Chanarin-Dorfman syndrome. Diagnostic suspicion was confirmed by genetic sequencing. Conclusions Chanarin-Dorfman syndrome is characterized by a mutation in the CGI-58 gene. This gene is involved in the catabolism of long-chain triglycerides stored in cytoplasmic lipid droplets. Jordans’ anomaly is a congenital alteration characterized by the presence of multiple vacuoles in the granulocytic series due to defective lipid metabolism. In this syndrome, long-chain triglycerides build up in tissues, thereby causing dermatological manifestations that are controllable through diet.

Single cell and spatial transcriptomics highlight the interaction of club-like cells with immunosuppressive myeloid cells in prostate cancer

Prostate cancer treatment resistance is a significant challenge facing the field. Genomic and transcriptomic profiling have partially elucidated the mechanisms through which cancer cells escape treatment, but their relation toward the tumor microenvironment (TME) remains elusive. Here we present a comprehensive transcriptomic landscape of the prostate TME at multiple points in the standard treatment timeline employing single-cell RNA-sequencing and spatial transcriptomics data from 120 patients. We identify club-like cells as a key epithelial cell subtype that acts as an interface between the prostate and the immune system. Tissue areas enriched with club-like cells have depleted androgen signaling and upregulated expression of luminal progenitor cell markers. Club-like cells display a senescence-associated secretory phenotype and their presence is linked to increased polymorphonuclear myeloid-derived suppressor cell (PMN-MDSC) activity. Our results indicate that club-like cells are associated with myeloid inflammation previously linked to androgen deprivation therapy resistance, providing a rationale for their therapeutic targeting.

Resolvin E1 and Inhibition of BLT2 Signaling Attenuate the Inflammatory Response and Improve One-Lung Ventilation-Induced Lung Injury

ABSTRACT Introduction One-lung ventilation (OLV) is a prevalently used technique to sustain intraoperative pulmonary function. Resolvin E1 (RvE1), a specialized pro-resolving lipid mediator, accelerates the resolution of inflammation in the lungs. However, its therapeutic effects on OLV-induced lung injury remain unclear. Methods We initially developed an OLV rat model and treated it with RvE1. Subsequently, we assessed the wet/dry ratio of the lung tissue, performed hematoxylin and eosin staining, and calculated the ratio of polymorphonuclear cells to white blood cells in the bronchoalveolar lavage fluid. Additionally, we assessed apoptosis, inflammatory factor levels, and lung permeability in the rat lung tissues in the RvE1 treated and untreated groups and explored the molecular mechanisms mediated by RvE1. Results Our results indicated that RvE1 alleviated lung injury and inflammation and improved lung tissue apoptosis and permeability in OLV rats. Moreover, RvE1 suppressed the expression of the BLT1/2 signaling pathway and its ligands. The use of BLT2 and BLT1 inhibitors (LY255283 and U-75302, respectively) enhanced RvE1’s anti-inflammatory effects and reduced lung injury. Furthermore, synergistic treatment with the BLT2 inhibitor and RvE1 provided grater benefits by more effectively inhibiting the NF-kB, p38 MAPK, and ERK pathways. Discussion RvE1 and the inhibition of BLT2 signalling reduce the inflammatory response and mitigate OLV-induced lung injury. These findings suggest a novel therapeutic pathway for managing OLV-related complications.

Cytokine profile, differential somatic cell count, and oxidative status of Italian Mediterranean buffalo milk affected by the temperature-humidity index.

In the context of climate change, there has been an increased interest in improving management practices for animals genetically adapted to extreme environmental conditions, such as buffaloes. The temperature-humidity index (THI) is used to determine the severity of heat stress in livestock. This study aimed to evaluate the cytokine profile, oxidative staus, differential somatic cell count (DCC), and the surface expression and activity of myeloperoxidase (MPO) in the somatic cells (SCs) of buffalo. Milk samples (n = 216) were collected from the spring to summer season under three different THI classes (THI < 72; ≤72 THI < 76, and THI ≥ 76). The cytokine profile was determined using ELISA, and the expression of DSCC and MPO was determined by flow cytometry. MPO activity was performed on SC extracts using a specific ELISA kit. Oxidative status was determined by the antioxidant/oxidant balance combining the free radical scavenging activity levels, and reactive oxygen and nitrogenous species. The results on the cytokine profile showed that at the THI ≥ 76 the levels of both IL-10 and IFN-γ were highest. IL-1β secretion was lower at the THI < 72 than at the THI values ranging from ≤72 THI < 76. Higher levels of both TNF-α and IL-12 were registered in both THI < 72 and THI ≥ 76 classes. The level of IL-4 was higher in the THI ≥ 76 class than in the ≤72 to <76 range. Data on DCC showed a decrease in the percentage of macrophages and lymphocytes as the THI increased from the ≤72 to <76 range to THI ≥ 76. Furthermore, the highest percentage of polymorphonuclear leukocyte (PMNLs) was registered in both ≤72 to <76 and THI ≥ 76 classes. The MPO activity and surface expression on SC were lower at a THI above 76, which could be associated with an absence of inflammation. A condition of oxidative imbalance was registered as demonstrated by the lower levels of antioxidant/oxidant balance along with increasing THI. Present data demonstrated that buffaloes were able to modulate the alteration of immune response activated by heat stress throughout a series of cross-linked mechanisms involving cytokine networks, different somatic cell distribution, and oxidative status.

The PdxR-PdxKU locus involved in vitamin B6 salvage is important for group A streptococcal resistance to neutrophil killing and survival in human blood.

Streptococcus pyogenes (Group A Streptococcus, GAS) is a Gram-positive bacterium that inflicts both superficial and life-threatening diseases on its human host. Analysis of fitness using a transposon mutant library revealed that genes predicted to be involved in vitamin B6 acquisition are associated with fitness in whole human blood. Vitamin B6 is essential for all life and is important for many cellular functions. In several streptococcal species, it has been shown that mutants in B6 acquisition exhibited reduced virulence phenotypes and were attenuated during infection. In GAS, B6 acquisition is believed to be controlled by the pdxR-pdxKU locus, where PdxR is a positive regulator of pdxKU, which encodes for a B6-substrate kinase and permease, respectively. Mutants in the regulator (ΔpdxR) and salvage machinery (ΔpdxKU) both exhibited modest growth defects when grown in oxygenated conditions with limited vitamin B6 precursors. ∆pdxR and ∆pdxKU mutants also exhibited an impaired ability to survive when challenged with whole human or mouse blood. This defect was characterized by reduced survival in the presence of human neutrophil-like HL60s, primary polymorphonuclear leukocytes, and antimicrobial peptide LL-37. Promoter analysis showed that PdxR is an autoregulator and activated pdxKU in the absence of B6. Interestingly, ∆pdxR and ∆pdxKU mutants were not attenuated in mouse models of infection, suggesting a species-specific impact on virulence. Overall, it appears that pdxR-pdxKU is associated with GAS vitamin B6 metabolism as well as pathogen survival during encounters with the human innate immune system.IMPORTANCEBacterial pathogens such as Streptococcus pyogenes (Group A Streptococcus, GAS) must be able to obtain needed nutrients in their human host. Vitamin B6 or pyridoxal 5' phosphate is essential for all life and is important for many cellular functions. In other streptococcal pathogens, B6 acquisition has been shown to be important for their ability to cause disease. Here, we show that loss of the putative vitamin B6 salvage pathway locus pdxR-pdxKU affects GAS pathogenesis when encountering innate immune responses from phagocytic neutrophils and antimicrobial peptides within the host. pdxR-pdxKU may contribute to oxygen tolerance through B6; however, there appear to be other mechanisms for salvaging vitamin B6. Overall, pdxR-pdxKU is associated with GAS resistance to the human innate immune response and oxygen tolerance and contributes modestly to B6 metabolism.

Neutrophils enhance the clearance of systemic amyloid deposits in a murine amyloidoma model.

Amyloid-specific antibodies have been shown to opsonize and enhance amyloid clearance in systemic amyloidosis mouse models. However, the immunological mechanisms by which amyloid is removed have not been clearly defined. Previous reports from preclinical in vivo studies suggest polymorphonuclear cells (i.e., neutrophils) can affect amyloid removal. Therefore, we sought to analyze how neutrophils may contribute to the clearance of human AL amyloid extracts, using a murine amyloidoma model. Immunocompromised nude mice injected subcutaneously with patient-derived AL amyloid extract (generating a localized "amyloidoma") were used to circumvent confounding factors contributed by the adaptive immune system and served as the model system. Two representative AL amyloid extracts were used, ALλ(CLA), which is refractory to clearance, and ALκ(TAL), which is readily cleared in mice. Neutrophil recruitment to the amyloid masses, cellular activation, and propensity to engulf amyloid were assessed. Immunophenotyping of amyloidomas from animals implanted with 2 mg of either ALλ or ALκ revealed that more neutrophils were recruited to ALκ amyloid masses as compared to the ALλ material, which was generally devoid of neutrophils. Ex vivo analyses indicated neutrophils do not efficiently phagocytose amyloid directly. However, histological evaluation of the ALκ amyloidoma revealed the abundant presence of neutrophil extracellular traps, which were absent in the ALλ amyloidomas. Using neutrophil depletion experiments in mice, we determined that mice devoid of neutrophils cleared the human amyloid lesions less efficiently. Moreover, mice devoid of neutrophils also had significantly reduced intra-amyloid expression of inflammatory cytokines. Neutrophils may not directly mediate amyloid clearance through phagocytosis; however, these cells can be stimulated by the amyloid and may function to facilitate phagocytosis and amyloid clearance by professional phagocytes (e.g., macrophages).

Tryptophan-immunoadsorption plasmapheresis regulates polymorphonuclear-myeloid-derived suppressor cells and pro-inflammatory cytokines.

Immunoadsorption plasmapheresis (IA) has been reported to have immunoregulatory effects, in addition to the removal of autoantibodies. This study aimed to investigate the effects of IA on the proportion of myeloid-derived suppressor cells (MDSCs) that potentially suppress autoimmune responses and regulate immunity. The study included 21 patients with autoimmune neurological diseases and 8 healthy participants. We measured polymorphonuclear (PMN)-MDSCs (CD14-CD11b+CD33+) and inflammation-related mediators before and after a single session of tryptophan-IA. We also investigated whether an increase in PMN-MDSCs after initial IA was a predictor of clinical efficacy in nine patients with myasthenia gravis based on the Quantitative Myasthenia Gravis score. For a total of 36 times of IA procedures, the number of PMN-MDSCs significantly increased after IA. Interleukin-10, monocyte chemoattractant protein-1 and macrophage inflammatory protein-1β levels showed significant increases after IA. Despite similar severity at admission, the Quantitative Myasthenia Gravis scores at discharge were significantly lower in the group in which IA increased PMN-MDSCs to a level of 20% of peripheral blood mononuclear cells or more. Tryptophan-IA regulates PMN-MDSCs and pro-inflammatory cytokines, possibly leading to suppression of autoimmune responses and tissue damage in neuroimmunological disorders.

Egfl6 promotes ovarian cancer progression by enhancing the immunosuppressive functions of tumor-associated myeloid cells.

Tumor-associated macrophages (TAMs) and myeloid-derived suppressor cells (MDSCs) play a critical role in resistance to immunotherapy. In this study, we identified epidermal growth factor-like 6 (Egfl6) as a regulator of myeloid cell functions. Our analyses indicated that Egfl6, via binding with β3 integrins and activation of p38 and SYK signaling, acts as a chemotactic factor for myeloid cell migration and promotes their differentiation toward an immunosuppressive state. In syngeneic mouse models of ovarian cancer (OvCa), tumor expression of Egfl6 increased the intratumoral accumulation of polymorphonuclear (PMN) MDSCs and TAMs and their expression of immunosuppressive factors, including CXCL2, IL-10, and PD-L1. Consistent with this, in an immune 'hot' tumor model, Egfl6 expression eliminated response to anti-PD-L1 therapy, while Egfl6 neutralizing antibody decreased the accumulation of tumor-infiltrating CD206+ TAMs and PMN-MDSCs and restored the efficacy of anti-PD-L1 therapy. Supporting a role in human tumors, in human OvCa tissue samples, areas of high EGFL6 expression colocalized with myeloid cell infiltration. scRNA-Seq analyses revealed a correlation between EGFL6 and immune cell expression of immunosuppressive factors. Our data provide mechanistic insights into the oncoimmunologic functions of EGFL6 in mediating tumor immune suppression and identified EGFL6 as a potential therapeutic target to enhance immunotherapy in patients with OvCa.

Fruquintinib Combined With PD-1 Inhibitors for the Treatment of the Patients With Microsatellite Stability Metastatic Colorectal Cancer: Real-world Data

BackgroundProgrammed death-1 (PD-1) or programmed death-ligand 1 (PD-L1) inhibitors have shown limited effectiveness in patients with microsatellite stable (MSS) metastatic colorectal cancer (mCRC). Combining anti-angiogenesis inhibitors with PD-1 inhibitors has the potential to reverse the immunosuppressive tumor microenvironment, synergistically enhancing the antitumor immune response in MSS mCRC. The goal is to present real-world data that prove the clinical efficacy and safety of fruquintinib combined with PD-1 inhibitors in MSS mCRC. Materials and methodsWe conducted a real-world retrospective study in patients with MSS mCRC who received treatment with fruquintinib combined with PD-1 inhibitors between May 2019 and March 2023 in our center. Results77 patients with MSS mCRC received fruquintinib combined with PD-1 inhibitors. In total, 5.2% of patients (4/77) achieved a partial response (PR), while 50.6% (39/77) had a stable disease (SD). Notably, three lesions achieving PR were all lung metastases and the overall disease control rate (DCR) reached 55.8% (43/77). Median progression-free survival (PFS) and overall survival (OS) reached 5.1 months (95% CI:3.6-6.7) and 14.6 months (95% CI:9.6-15.6), respectively. Multivariate Cox analysis showed that prior treatment without vascular endothelial growth factor (VEGF) inhibitors was significantly associated with PFS and OS (p<0.05). Further analysis indicated that total- or polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) significantly decreased after treatment (p=0.039), especially in the PR/SD group (p=0.003). Most adverse events included abdominal pain, rash, edema, diarrhea, and immunotherapy-associated hypothyroidism, yet symptoms were controllable. ConclusionOur results provided additional evidence that patients with MSS mCRC could benefit from the combination of fruquintinib and PD-1 inhibitors, especially those with lung metastases or without prior treatment with VEGF inhibitors. The detection of MDSCs may be an immune indicator for predicting of the combined therapy.

Frequency of Asymptomatic Spontaneous Bacterial Peritonitis in Patients with Chronic Liver Disease and Ascites

Chronic liver disease (CLD) contributes significantly to morbidity and mortality around the world. In Pakistan, it is due mainly to infections of Hepatitis B and C. One of the complications of CLD, which is often important for such patients, involves an infection of spontaneous bacterial peritonitis. Asymptomatic spontaneous bacterial peritonitis commonly develops in asymptomatic patients with ascites and is therefore preventable when it develops early. This study will determine the incidence of asymptomatic SBP in patients with CLD and ascites, providing potentially valuable data to support screening protocols for early intervention and improved outcomes. It was a cross-sectional study carried out for six months in the Gastroenterology Department of Sheikh Zayed Hospital, Lahore. A total of 121 CLD patients with ascites, meeting particular inclusion criteria, were assessed. The cases were categorized as asymptomatic SBP if the cultures were positive with more than 10⁵ colonies or more than 250/mm³ polymorphonuclear cells, without fever, pain, or tenderness. The patients were stratified according to age, gender, and Child-Pugh classification. The data were analyzed using SPSS version 24, along with the Chi-square test for correlation between the Child-Pugh class and SBP.The proportion of asymptomatic SBP was at 14.8 % among 121 patients. The patients were older: 66.1%, were mostly more than 50 years old, with a fair sex distribution ratio of 52.1 females against 47.9 males. Patients classified into Child-Pugh Class C constituted the highest proportions of patients with SBP cases of 14.8 % and significant association at p=0.000. Asymptomatic SBP in 14.8% of the patients with CLD and ascites forms the justification for routine screening on hospital admission. Critical importance is placed on detecting asymptomatic SBP because this leads to earlier intervention and a potential reduction in the risk of major complications, particularly in patients classified as Child Class C. Keywords- Frequency, Asymptomatic, Spontaneous Bacterial Peritonitis, Patients Chronic Liver Disease, Ascites

Adverse effects of CXCR2 deficiency in mice reared under non-gnotobiotic conditions

The family of pro-inflammatory and pro-angiogenic chemokines including Interleukin-8 (IL-8, aka CXCL8) and its homologues (CXCL1,2,3,5,6, and 7) exhibit promiscuous binding and activation of several G-protein-coupled receptors (i.e., CXCR2, CXCR1, and the atypical chemokine receptor (ACKR1)). A high proportion of their biological activity is attributed to CXCR2 activation, thus many CXCR2 inhibitors are in clinical trials for several chronic diseases. However, CXCR2 inhibition is often only investigated acutely in these trials or in Cxcr2−/− mice grown in gnotobiotic conditions. Since humans do not live in germ-free environments, our first goal is to highlight novel retinal and systemic observations in Cxcr2−/− mice grown in non-gnotobiotic conditions that suggest potential harmful consequences of long-term CXCR2 deficiency or blockade. Beyond confirmation of circulating blood/immune cell-related phenotypes, we report novel findings in Cxcr2−/− mice including: (1) delayed dye transit to the retinal vasculature, (2) alterations in the density and distribution of retinal vessels, astrocytes and microglia, (3) decreased electroretinogram a- and b-wave amplitudes, (4) reduced visual acuity, and (5) increased polymorphonuclear cell accumulation in vascular lumina abutting venular walls in the retina and in vital non-ocular tissues (lung and liver). Furthermore, PheWAS of CXCR2 CXCR1, and ACKR1 gene variants using data from UK Biobank participants suggest clinical associations with both retinal and vascular disease phenotypes. We conclude that chronic CXCR2 deficiency in mice contributes to functional damage to the retina and that the long-term safety of CXCR1/2 inhibitors designed for chronic use in humans should be explored before clinical adoption to safeguard sight and overall vascular health.

IN VITRO EVALUATION OF IMMUNOSTIMULANT ACTIVITY OF ISOLATED EMBELIN FROM EMBELIA RIBES

Traditional medicinal plants have long been used in people’s healthcare regimens to treat a wide range of ailments. The ancient medicinal plant Embelia ribes Burm. (E. ribes) has potential for a wide range of pharmacological activities that are linked to embelin, a natural dihydroxy benzoquinone. E. ribes is reported to have immunomodulatory activity, considering which was the foundation to sketch an immunomodulatory study of the isolated embelin from it. Analytical assessment confirmed the isolated embelin by FTIR, UV-Visible spectrophotometry and DSC when compared with the standard. Embelin showed immunostimulant activity in the concentration range of 160 µg mL-1 to 240 µg mL-1. The in vitro study on human PMN cells, observations of phagocytosis of Candida albicans, and NBT reduction assay explain that embelin has an affinity towards PMN cells.

Histopathological Description of Mouse Liver in a Sepsis Model Infected with Escherichia coli Treated with Paederia foetida L. Leaf Extract for Sepsis Prevention

The leaf of Paederia foetida L. is one type of medicinal plant that can be used as a preventive medicine for sepsis. This plant contains secondary metabolites such as alkaloids, flavonoids, triterpenoids, saponins, and other active compounds. The objective of this study was to determine the histopathological description of the liver in a mouse sepsis model infected with E. coli, with the administration of Paederia foetida L. leaf extract for sepsis prevention, and to ascertain the influence and effective dosage of the leaf extract as a preventive measure against liver histopathology in the sepsis model induced by E. coli. The method employed was a Completely Randomized Design (CRD). The study used 24 male white mice divided into 6 (six) groups. Data analysis was conducted using One Way ANOVA. The results of the study revealed the histopathological profile of liver cell degeneration in group PI (100mg/kg BW) at 20.79%±0.03, group PII (200mg/kg BW) at 21.63%±0.02, and group PIII (500mg/kg BW) at 9.08%±0.02. Necrosis rates were observed in group PI (100mg/kgBW) at 22.62%±0.04, group PII (200mg/kg BW) at 17.63%±0.02, and group PIII (500mg/kg BW) at 6.05%±0.02. The presence of polymorphonuclear leukocytes (PMN) was detected in group PI (100mg/kgBW) at 39.56%±0.03, group PII (200mg/kgBW) at 28.05%±0.02, and group PIII (500mg/kg BW) at 18.45%±0.03. The test results showed a significant effect of P. foetida L. leaf extract as a preventive measure against liver histopathology in the mouse sepsis model infected with E. coli, with significant values for necrosis (p=0.000), cell degeneration (p=0.000), and PMN (p=0.000). The most effective dosage of P. foetida L. leaf extract as a preventive measure against liver histopathology in the mouse sepsis model infected with E. coli was the dosage used in group PIII (500mg/kgBW).

Impact of Nitric Oxide on Polymorphonuclear Neutrophils' Function.

There is increasing evidence that nitric oxide (nitrogen monoxide, NO) significantly influences immune cellular responses, including those from polymorphonuclear leukocytes (PMNs). The aim of this study was to examine a possible effect of NO on PMNs' function (chemotaxis, production of reactive oxygen species (ROS), and NETosis) using live cell imaging. Moreover, we investigated PMN surface epitope and neutrophil oxidative burst under the influence of NO by flow cytometric analysis. Whole blood samples were obtained from healthy volunteers, and PMNs were isolated by density centrifugation. Live cell imaging using type I collagen matrix in µSlide IBIDI chemotaxis chambers was conducted in order to observe N-formyl-L-methionyl-L-leucyl-phenylalanine (fMLP)-stimulated PMN chemotaxis, ROS production, and NETosis. In the test group, NO was continuously redirected into the climate chamber of the microscope, so the chemotaxis chambers were surrounded by NO. The same experimental setup without NO served as a control. In addition, isolated PMNs were incubated with nitrogen monoxide (NO) or without (the control). Subsequently, flow cytometry was used to analyze neutrophil antigen expression and oxidative burst. Our live cell imaging results demonstrated a migration-promoting effect of NO on PMNs. We observed that in the case of prior stimulation by fMLP, NO has no effect on the time course of neutrophil ROS production and NET release. However, flow cytometric analyses demonstrated an increase in ROS production after pretreatment with NO. No NO-dependent differences for the expression of CD11b, CD62L, or CD66b could be observed. We were able to demonstrate a distinct effect of NO on PMNs' function. The complex interaction between NO and PMNs remains a major research focus, as the exact mechanisms and additional influencing factors remain elusive. Future studies should explore how varying NO concentrations and the timing of NO exposure relative to PMN activation affect its influence.