Polymorphonuclear Leukocyte Count Research Articles

Pinitol attenuates LPS-induced pneumonia in experimental animals: Possible role via inhibition of the TLR-4 and NF-κB/IκBα signaling cascade pathway.

Pneumonia is a chronic disorder of the respiratory system associated with worsening quality of life and a significant economic burden. Pinitol, a plant cyclic polyol, has been documented for immune-inflammatory potential. The aim of present investigation was to evaluate the potential and possible mechanism of action of pinitol against lipopolysaccharide (LPS)-induced pneumonia in the experimental animal model. Pneumonia was induced in Sprague-Dawley rats by intratracheal administration of LPS (2 mg/kg). Animals were treated with either vehicle or dexamethasone or pinitol (5 or 10 or 20 mg/kg). Potential of pinitol against LPS-induced pulmonary insult was assessed based on behavioral, biochemical, molecular, and ultrastructural studies. Intratracheal instillation of LPS induced significant (P < .05) inflammatory infiltration inbronchoalveolar lavage fluid (BALF) and lung tissue reflected by elevated pleural effusion volume, lung edema, BALF polymorphonuclear leukocytes count and lung myeloperoxidase levels, which was attenuated by pinitol (10 and 20 mg/kg) administration. Pinitol also markedly (P < .05) inhibited LPS-induced alterations in electrocardiographic, hemodynamic changes, right ventricular, and lung function tests. The LPS-induced downregulated nuclear factor erythroid 2-related factor 2 (Nrf-2) and heme oxygenase-1 (HO-1), whereas upregulated transforming growth factor-β (TGF-β), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3), and inducible nitric oxide synthase (iNOs) lung messenger RNA expressions were significantly (P < .05) inhibited by pinitol. Western blot analysis suggested pinitol markedly (P < .05) decreased nuclear factor-κB (NF-κB), inhibitor of nuclear factor κB (IkBα), toll-like receptor 4 (TLR-4), and cyclooxygenase-II (COX-II) protein expressions in the lung. These findings were further supported by histological and ultrastructural analyses of lung tissue that showpinitol significantly (P < .05) ameliorates LPS-induced aberrations in lung tissue. In conclusion, pinitol attenuated LPS-induced pneumonia via inhibition of TLR-4 to downregulate the NF-κB/IκBα signaling cascade and thus ameliorated the production of proinflammatory cytokines (TNF-α, ILs, NLRP3, and TGF-β), inflammatory mediators (COX-II and iNOs) and elevated oxidative stress (Nrf-2 and HO-1).

Predictors of immediate and short-term mortality in spontaneous bacterial peritonitis.

There is scarce data from the Indian subcontinent on the outcomes following spontaneous bacterial peritonitis (SBP). To study the immediate (within 30days) and short-term mortality (31-90days) associated with SBP and to determine the predictors of the same. This prospective observational study was done among patients with liver cirrhosis who underwent paracentesis. Patient data included age, gender, co-morbidity, cirrhosis-related complications, model of end-stage liver disease (MELD), and Child-Turcotte-Pugh (CTP) scores. SBP was diagnosed based on ascitic fluid polymorphonuclear leukocyte count > 250/mm3 with or without ascitic fluid culture positivity. Of the 870 patients with cirrhosis and ascites registered during the study period, 610 fulfilled the criteria for inclusion. Altogether, 122 patients with SBP were identified: 52 (42.6%) died, 40 (32.8%) survived without liver transplant, and 30 (24.6%) underwent liver transplantation within 3months. Thirty-two patients (26.2%) were blood culture posi tivefor bacteria and 7 (5.7%) demonstrable bacterialgrowth in ascitic fluid. Blood culture positivity was significantly higher in the group with immediate mortality (p < 0.0001) and was also significantly associated (p 0.005) with mortality at 3months. Nearly two-fifths (42.6%) of the study cohort died within 3months of an episode of SBP. Four-fifths of these patients died within 30days. Blood culture positivity was significantly associated with immediate and short-term mortality.

The diagnostic accuracy of ascitic calprotectin for the early diagnosis of spontaneous bacterial peritonitis: systematic review and meta-analysis.

Spontaneous bacterial peritonitis (SBP) is a common complication in patients with cirrhosis and has an incidence of up to 30% in hospitalized patients. Importantly, it may raise their mortality rate up to 30%. Hence, a delayed diagnosis is associated with poor prognosis. This systematic review aims to assess the diagnostic accuracy of ascitic fluid calprotectin for the early diagnosis of SBP in patients with ascites. This study was conducted in accordance with the PRISMA statement. A systematic literature search was conducted from inception to February 2020 in the following electronic bibliographic databases: MEDLINE, Scopus, The Cochrane Library and OpenGrey. Quality Assessment of Diagnostic Accuracy Studies tool was used to assess risk of bias. Ten studies were included in the qualitative and quantitative synthesis. Hierarchical summary receiver operating characteristic curves were plotted and the summary sensitivity of a positive ascitic fluid calprotectin assessment to detect SBP was 93% [95% confidence interval (CI) 90-95%] while the summary specificity was 89% (95% CI 80-95%), irrespectively of the method used. The positive likelihood ratio and negative likelihood ratio of the test were 8.7 (95% CI 4.4-17.1) and 0.08 (85% CI 0.06-0.12). All studies showed positive correlation between ascitic calprotectin and polymorphonuclear (PMN) leukocyte count. Ascitic calprotectin appears to be an excellent alternative to PMN leucocyte count of ≥250 cells/mm3 for the diagnosis of SBP with much faster time to diagnosis. Owing to its substantially high negative predictive value, the test can accurately exclude SBP avoiding unnecessary antibiotics in suspected patients.

Kortikosteroid ve Antihistaminik Enjeksiyonların Sıçan Mesane Dokusunda Soğuk Kaynaklı Stres Üzerine Etkileri: Deneysel Bir Çalışma

Objective: Cold is a stress-inducing factor that can cause changes in bladder function in various ways. The present study is intended to investigate the effects of corticosteroid and antihistaminic treatment on acutely and chronically induced cold stress.Material and Methods: Forty-two female rats were randomly divided into seven groups as follows: Control group; Acute cold-stress (ACS) group; ACS+ corticosteroid (CORT) group; ACS+CORT + Antihistaminic group; Cronic cold-stres (CCS) group; CSS+CORT group; CSS+CORT+Antihistaminic group. On the 15th day after these treatments, bladders of the rats were harvested for histopathological examinations under general anesthesia and fixed with 10% neutral buffered formaline. Hematoxylin and eosin and toluidine blue stainings were performed.Results: The comparison based on mast cell count yielded the highest value in the CCS group in comparison to the control group. The lowest value was harvested in the CCS + CORT group. The comparison between the CCS groups revealed the highest polymorphonuclear leucocyte (PNL) values in the lamina propria in the CCS group, whereas the CORT and CORT + Antihistaminic treatments were found to have significantly decreased the PNL values in the lamina propria. While the PNL counts in the epithelium were high in the ACS and CCS groups, the results in the ACS and CCS groups that were treated with CORT and/or antihistaminic were revealed to be similar with those in the control group. It was discovered that antihistaminic injection in addition to CORT decreased the lymphocyte counts in epithelium in CCS more efficiently than CORT alone did.Conclusion: The present research revealed that corticosteroid treatment reduced inflammatory cell infiltration and decreased mast cell count. A more evident amelioration was observed particularly in chronic cold stress.

The pattern of microbial flora in Spontaneous bacterial peritonitis in Cirrhotic patients

Background: Spontaneous bacterial peritonitis (SBP) is the bacterial infection of ascitic fluid in the absence of other apparent causes. It is the most prevalent and serious infection in cirrhosis and leads to significant morbidity and mortality. The frequency of SBP in hospitalized patients with cirrhosis varies from 7 to 23% in the west and it is around 33% in Pakistan. Our study aim was to determine the frequency of microbial organisms isolated from spontaneous bacterial peritonitis in cirrhotic patients. Material and Methods: We did a cross-sectional study at the Department of Medicine, Fauji Foundation Hospital Rawalpindi for 6 months. Cirrhosis was diagnosed on the bases of clinical, biochemical and ultrasonography findings. Peripheral blood samples were obtained for the evaluation of complete blood cell count, polymorphonuclear leukocyte count, albumin and coagulation parameters. At the same time, the patients underwent paracentesis and a 20ml ascitic fluid sample was taken for culture sensitivity along with routine biochemical examination from the pathology department lab at our hospital. Results: A total of 150 patients were enrolled according to the inclusion criteria of the study. The mean age (yrs) of patients was 32.3+9.31. The frequency and percentages of male and female patients were 70 (46.7) and 80 (53.3) respectively. Frequency and percentages of E Coli and streptococcal pneumonia were 94 (62.7) and 34 (22.7) respectively, whereas frequency and percentages of staph aureus and Klebsiella were 22 (14.7). Conclusion: SBP is quite common in patients with cirrhosis. It should be suspected in all such cases presenting with typical or atypical features. E Coli was the most common causative organisms in our study but gram-positive organisms were quite frequent as well. Keywords: SBP, Cirrhosis, Peritonitis.

Ascitic calprotectin as a useful marker in the diagnosis of spontaneous bacterial peritonitis in adults

BackgroundAscitic fluid polymorphonuclear leucocyte count (PMN) is known to be the gold standard for spontaneous bacterial peritonitis (SBP) diagnosis. The aim of this work was to assess ascitic calprotectin for SBP diagnosis. Serum C-reactive protein (CRP), high sensitivity C-reactive protein (hsCRP), nitrous oxide, ascitic PMN, ascitic leucocyte esterase and ascitic calprotectin were measured.ResultsThe average age of our patients was 55.25 ± 7.89 years, mostly males (n = 51, 63.8%), anti-HCV antibodies were positive in (n = 61, 76.3%). Sixty-four patients (80%) were Child-Pugh C and their average MELD was 24.29 ± 8.06. Patients with SBP had statistically significant higher median MELD score (26.5 vs. 19) and higher average Child-Pugh score (12.18 ± 1.74 vs. 10.5 ± 1.97). Forty patients had SBP and 40 patients were without SBP. Both the serum and ascitic nitrous oxide did not differ statistically between patients with and without SBP. In contrast, patients with SBP had higher median serum CRP (49 vs. 12 mg/dL), hsCRP (58,000 vs. 23,750 ng/dL) and ascitic calprotectin (7.57 vs. 1.1 ng/mL). The ascitic leucocyte esterase test was positive in 95% of SBP patients in contrast to 2.5% patients without SBP. Ascitic calprotectin >2 ng/mL had 90% sensitivity, 92.5% specificity, 92.3% positive predictive value and 90.2% negative predictive value. MELD, CRP, hsCRP and ascitic calprotectin are independent predictors of SBP.ConclusionAscitic calprotectin is a useful marker for SBP diagnosis.

The value of delta neutrophil index in neonatal sepsis diagnosis, follow-up and mortality prediction.

The complete blood cell count (CBC) and peripheral blood smear were the most commonly ordered tests for the diagnosis of neonatal sepsis. Delta neutrophil index (DNI) shows leucocyte differentiation and calculated while CBC is performed. We aimed to evaluate the value of DNI in neonatal sepsis. DNI was measured with Siemens Advia 2120 and 2120i devices. DNI was calculated as (neutrophil and eosinophil count in myeloperoxidase channel)-(polymorphonuclear leucocyte count in nuclear lobularity channel). Study population included 141 and 87 neonates in sepsis (110 proven, 31 clinical) and control groups. Demographic characters were similar between groups. Proven sepsis group had lower birthweight and higher late-onset sepsis rate than clinical sepsis and control groups. Median DNI (16.3 vs 1,4) and CRP (6.8 vs 0,03 mg/dl) were significantly higher in sepsis group. Proven sepsis group had significantly higher DNI level than clinical sepsis group (20.8 vs 9.1). Cut-off level of DNI was 4.6 with 85% sensitivity and 80% specificity. Cut-off level of CRP was 0.58 mg/dl with 81% sensitivity and 82% specificity. Combination of DNI and CRP gave 98% sensitivity and 76% specificity. Mortality rate in sepsis group was 39%. Median DNI level in patients with mortality was significantly higher (30.1 vs 9.6). Cut-off level of DNI for mortality prediction was 16.1 with 75% sensitivity and 65% specificity. Follow-up levels of DNI was significantly decreased in 6-10 days to normal levels (16.3 to 4.2). DNI was found to be useful in the diagnose, follow-up and mortality prediction of neonatal sepsis without extra blood to CBC.

An inflammatory profile linked to increased suicide risk

BackgroundSuicide risk assessments are often challenging for clinicians, and therefore, biological markers are warranted as guiding tools in these assessments. Suicidal patients display increased cytokine levels in peripheral blood, although the composite inflammatory profile in the subjects is still unknown. It is also not yet established whether certain inflammatory changes are specific to suicidal subjects. To address this, we measured 45 immunobiological factors in peripheral blood and identified the biological profiles associated with cross-diagnostic suicide risk and depression, respectively. MethodsSixty-six women with mood and anxiety disorders underwent computerized adaptive testing for mental health, assessing depression and suicide risk. Weighted correlation network analysis was used to uncover system level associations between suicide risk, depression, and the immunobiological factors in plasma. Secondary regression models were used to establish the sensitivity of the results to potential confounders, including age, body mass index (BMI), treatment and symptoms of depression and anxiety. ResultsThe biological profile of patients assessed to be at increased suicide risk differed from that associated with depression. At the system level, a biological cluster containing increased levels of interleukin-6, lymphocytes, monocytes, white blood cell count and polymorphonuclear leukocyte count significantly impacted suicide risk, with the latter two inferring the strongest influence. The cytokine interleukin-8 was independently and negatively associated with increased suicide risk. The results remained after adjusting for confounders. LimitationsThis study is cross-sectional and not designed to prove causality. DiscussionA unique immunobiological profile was linked to increased suicide risk. The profile was different from that observed in patients with depressive symptoms, and indicates that granulocyte mediated biological mechanisms could be activated in patients at risk for suicide.

The value of ascitic fluid calprotectin and calprotectin-to-albumin ratio in the diagnosis and prognosis of spontaneous bacterial peritonitis

Introduction Spontaneous bacterial peritonitis (SBP) is a potentially fatal condition, characterized by infection of ascitic fluid (AF) in the absence of any intra-abdominal surgically treatable source of infection. It is the most frequent and severe complication of cirrhotic ascites. SBP is a condition that requires a high index of suspicion, rapid and accurate diagnosis, in addition to prompt and effective therapy. Aim The aim of this study was to evaluate AF calprotectin as a diagnostic marker in detecting SBP. In addition, we have evaluated AF calprotectin-to-albumin ratio in the diagnosis and prognosis of SBP. Patients and methods A total of 72 patients with cirrhotic ascites were included in this study. They were divided into two groups: SBP group included 50 patients with cirrhotic ascites and SBP diagnosed by presence of polymorphonuclear leukocyte count at least 250 cells/mm3 in AF with or without positive AF culture, and non-SBP group included 22 patients with cirrhotic ascites without evidence of SBP. All patients were subjected to complete clinical evaluation, laboratory investigations, diagnostic abdominal paracentesis, serum and AF C-reactive protein levels, which were assessed quantitatively, and AF calprotectin levels, which were measured by quantitative sandwich enzyme-linked immunosorbent assay. Results AF calprotectin was significantly elevated in patients with SBP in comparison with non-SBP patients (P&lt;0.001), with the best cutoff value for the detection of SBP (372 ng/ml) with a sensitivity, specificity, positive predictive value, negative predictive value, and an accuracy of 100% for each. Moreover, there was a positive correlation with total leukocytic count, polymorphonuclear leukocyte, and C-reactive protein in serum and AF. Moreover, calprotectin-to-albumin ratio was increased in SBP group versus non-SBP group (P&lt;0.001). Conclusion AF calprotectin can be used as a valuable marker in rapid diagnosis of SBP. Moreover, calprotectin-to-albumin ratio in ascites is useful in the diagnosis of SBP, as well as it provides prognostic information on short-term survival of patients with SBP with follow-up treatment.

Risk factors predicting nosocomial, healthcare-associated and community-acquired infection in spontaneous bacterial peritonitis and survival outcome.

Aim of the studyTo determine risk factors predicting nosocomial, healthcare-associated and community-acquired infection in spontaneous bacterial peritonitis (SBP) and survival outcome.Material and methodsThis prospective observational study included confirmed cases of cirrhosis with ascites requiring paracentesis, age > 18 years, either gender, any aetiology and Child-Turcotte-Pugh (CTP) stage, with or without cirrhosis-related complications. Patient data included age, gender, co-morbidity, model for end-stage liver disease (MELD) score, CTP score, cirrhosis-related complications, details of previous hospitalization, ascitic tapping and antibiotics instituted. SBP was diagnosed as ascitic fluid polymorphonuclear leucocyte count greater than 250/mm3 (0.25 × 109/l) and/or culture positivity for a single organism. Statistics – chi square test, Mann-Whitney U test, ANOVA, survival plot. A p value < 0.05 was statistically significant.Results610 cases fulfilled the criteria for inclusion. 122 (20%) patients had SBP: community-acquired SBP 37 (30.3%), nosocomial SBP 19 (16.5%) and healthcare-associated SBP 66 (54.5%). The majority were men (106; 86%) with median age of 51.5 (27-78) years. A significantly higher percentage of community-acquired SBP belonged to CTP class B. Thirtytwo and 7 patients respectively were blood and ascitic fluid culture positive. Significant nosocomial SBP were blood culture positive (p < 0.02). The most common isolates were E. coli followed by Klebsiella. Survival plot analysis at 3 months showed the worst survival for nosocomial SBP (p = 0.0009).ConclusionsPrevalence of SBP in our study was 20%, the majority with healthcare-associated SBP belonging to CTP C. Patients with nosocomial SBP had significant bacteremia with high mortality.

Update Review Article: Spontaneous Bacterial Peritonitis

Spontaneous bacterial peritonitis (SBP) is a frequent and severe complication in cirrhotic patients with ascites. To describe spontaneous bacterial peritonitis (SBP) in the context of currently accepted criteria for diagnosis, treatment and prevention. A review of SBP and its associated etiopathogenic factors is presented. Numerous studies on mechanisms of disease, bacteriology, epidemiology, diagnostic markers, and current guidelines for its diagnosis, treatment and prevention are discussed. Peritonitis in patients with ascites in the absence of secondary causes, such as perforation of a viscus, occurs primarily in patients with end-stage liver disease. Enteric organisms, mainly gram-negative bacilli, probably translocate to regional lymph nodes to produce bacteremia and seeding of ascitic fluid. Signs and symptoms of peritonitis are usually subtle. The ascitic fluid polymorphonuclear leukocyte count is the best determinant for early diagnosis and treatment of SBP. Third-generation cephalosporins such as cefotaxime are considered the drugs of choice for treatment, whereas quinolones such as norfloxacin are used to decrease recurrence. Despite increased awareness, early diagnosis, and prompt and effective antimicrobial therapy, SBP recurs frequently and is associated with a high mortality rate. Patients with SBP should be assessed for candidacy for liver transplantation.&#x0D; Keyword: Spontaneous bacterial peritonitis, Bacteriology, Epidemiology, Diagnostic markers, Cephalosporins

Assessment of calprotectin in ascitic fluid as a marker for spontaneous bacterial peritonitis diagnosis in cirrhotic patients

BackgroundSpontaneous bacterial peritonitis (SBP) is a serious infection of ascitic fluid in cirrhotic patients. High mortality associated with the delay in diagnosis and treatment. There is a need for an accurate and a rapid method for SBP diagnosis.ObjectivesWe aimed to evaluate ascitic fluid calprotectin as a diagnostic marker for SBP.Patients and methodsForty four cirrhotic patients were divided into two groups, non-SBP group: 22 patients with cirrhotic ascites without evidence of SBP and SBP group: 22 patients with cirrhotic ascites and SBP diagnosed by positive ascitic fluid bacterial culture and an increase in polymorphonuclear leukocytes (PMNLs) count in ascites (≥250 cells/mm3). Ascitic fluid calprotectin levels were measured using enzyme-linked immunosorbent assay.ResultsThere was a significant increase of ascitic fluid calprotectin, total leukocytic count, PMNLs, lactate dehydrogenase, and total protein in SBP group when compared to non-SBP group. There were significant positive correlations between white blood cell, ascitic fluid total leukocytic count, PMNLs, total protein, and model for end-stage liver disease score values and ascitic fluid calprotectin among SBP group. Ascitic fluid calprotectin with cutoff value 620 ng/ml, showed a sensitivity of 90.91% and a specificity of 95.45%, in diagnosis of SBP with positive predictive value 95.2% and negative predictive value 91.3%.ConclusionAscitic fluid calprotectin may be valuable in rapid diagnosis of SBP.

The Role of Polymorphonuclear Leukocyte Counts from Urethra, Cervix, and Vaginal Wet Mount in Diagnosis of Nongonococcal Lower Genital Tract Infection.

Objective The aim of this study was to evaluate whether the polymorphonuclear leukocyte (PMNL) inflammatory response in women with nongonococcal lower genital tract infection (LGTI) can be used to optimize criteria for syndromic treatment. Methods A cross-sectional study of 375 women attending the STI clinic in Oslo. Urethral, cervical, and vaginal specimens underwent microscopy for PMNLs. Chlamydia trachomatis (Ct) and other STIs were detected in the cervical/vaginal swabs and urine, using nucleic acid amplification test (NAAT). After excluding vulvovaginal candidiasis, genital herpes, and trichomoniasis, we correlated clinical and microscopic signs of inflammation with positive NAAT for Ct, mycoplasma genitalium (Mg), and Ureaplasma urealyticum (Uu) in a subgroup of 293 women. Results To predict a positive Ct, the combination of high cut-off urethritis (≥10 PMNLs/HPF) and microscopic cervicitis had a high specificity of 0.93, a PPV of 0.37, and a sensitivity of 0.35. LGTI criteria had low predicting values for Mg and Uu. Conclusion Including microscopic criteria for the diagnosis of LGTI gives better indication for presumptive antibiotic treatment than anamnestic and clinical diagnosis alone.

Gastric aspirate cytology as a screening tool for neonatal sepsis: a prospective study from a tertiary care centre

Background: Neonatal sepsis is a clinical syndrome of bacteremia characterized by systemic signs and symptoms of infection in the first 28 days of life. It is responsible for 30-50% of the total neonatal deaths each year in developing countries. Gastric aspirate cytology has been used for neonatal infection. The presence of more than five polymorphs per high power field co-relate with neonatal infection. The objective of the present study was to evaluate the utility of gastric aspirate cytology as a screening tool for neonatal sepsis and to determine the polymorphonuclear leukocytes count in smear of gastric aspirate and correlating it with culture proven sepsis.Methods: This prospective observational study was conducted from February 2017 to January 2018 at level III Neonatal intensive care Unit of Sri Venkata Sai Medical College and Hospital, Mahabubnagar, Telangana. A total of 108 neonates with risk factor and / or clinical features of sepsis were included in the study.Results: Out of 108 neonates, 40 were blood culture positive and 68 were culture negative. Gastric aspirate smear showed ≥5 polymorphs in 30 and &lt;5 polymorphs in 10 neonates with positive blood culture. Among blood culture negative cases, 20 had ≥5 polymorphs and 48 had &lt;5 polymorphs in gastric aspirate smear. Gastric aspirate culture was positive in 48 neonates and negative in 60 neonates. Of the 48 gastric aspirate positive neonates, 45 had ≥5 polymorphs and 3 had &lt;5 polymorphs in gastric aspirate smear. Similarly, among 60 gastric aspirate culture negative neonates, 55 had ≥5 polymorphs and 3 had &lt;5 polymorphs in gastric aspirate smear. This was statistically significant (P&lt;0.000001). Of 48 neonates with positive gastric aspirate culture, 30 had positive blood culture and 18 had negative blood culture.Conclusions: Gastric aspirate cytology is a good screening tool for neonatal sepsis added to a detailed perinatal history and clinical examination but does not completely substitute the present day available screening parameters.

Inhibition of insulin-like growth factor receptor-1 reduces necroptosis-related markers and attenuates LPS-induced lung injury in mice

Insulin-like growth factor-1 (IGF-1) levels are known to increase in the bronchoalveolar lavage fluid (BALF) of patients with acute respiratory distress syndrome. Herein, we investigated the role of IGF-1 in lipopolysaccharide (LPS)-induced lung injury. In LPS-treated cells, expressions of receptor-interacting protein 3 (RIP3) and phosphorylated mixed lineage kinase domain-like protein (MLKL) were decreased in IGF-1 receptor small interfering RNA (siRNA)-treated cells compared to control cells. The levels of pro-inflammatory cytokines including interleukin (IL)-1β, IL-6, IL-10, tumour necrosis factor-α, and macrophage inflammatory protein 2/C-X-C motif chemokine ligand 2 in the supernatant were significantly reduced in IGF-1 receptor siRNA-treated cells compared to control cells. In LPS-induced murine lung injury model, total cell counts, polymorphonuclear leukocytes counts, and pro-inflammatory cytokine levels in the BALF were significantly lower and histologically detected lung injury was less common in the group treated with IGF-1 receptor monoclonal antibody compared to the non-treated group. On western blotting, RIP3 and phosphorylated MLKL expressions were relatively decreased in the IGF-1 receptor monoclonal antibody group compared to the non-treated group. IGF-1 may be associated with RIP3-mediated necroptosis in vitro, while blocking of the IGF-1 pathway may reduce LPS-induced lung injuries in vivo.

Ascitic calprotectin as a diagnostic marker for spontaneous bacterial peritonitis in hepatitis C virus cirrhotic Egyptian patients

BackgroundThe gold standard for the diagnosis of spontaneous bacterial peritonitis (SBP) is a polymorphonuclear leukocyte (PMNL) count of 250/mm3or more. Accurate and early diagnosis of SBP is important to decrease the mortality and complications in patients with cirrhosis.AimsThe aim of this study was to evaluate the accuracy of ascitic fluid calprotectin as a diagnostic marker for the detection of SBP.Patients and methodsSeventy Egyptian patients with liver cirrhosis and ascites were enrolled; these patients were divided into two groups: 50 patients with SBP and 20 patients with no SBP on the basis of an elevated ascitic PMNL count of 250 cells/mm3 or more. Ascitic samples were examined for PMNL count, culture, chemistry, and calprotectin concentrations in all patients.ResultsCalprotectin levels in ascitic fluid were correlated significantly with PMNLs and significantly higher in patients with SBP than non-SBP (P<0.001), with the best cutoff value for the detection of SBP of 783 ng/ml with a sensitivity, a specificity, a positive predictive value and negative predictive value, and an accuracy of 90, 100, 100, 80, and 92.9%, respectively.ConclusionElevated ascitic calprotectin levels in cirrhotic patients are a diagnostic and reliable marker for the detection of SBP and considered a surrogate marker for PMNL.

Does ascitic fluid lactoferrin has a role in the diagnosis and follow up of spontaneous bacterial peritonitis in hepatitis C virus cirrhotic patients

Background Polymorphonuclear leukocyte (PMNL) count in the ascitic fluid (AF) is the gold standard method for the diagnosis of spontaneous bacterial peritonitis (SBP). Its measurement is routinely performed by traditional manual counting which is operator dependent and false-negative results may occur due to lysis of the leukocyte during transport. Aims The aims of the study were to assess the accuracy of AF lactoferrin and the best cutoff value for the diagnosis of SBP and also to compare its level before and after treatment to be used as a marker for follow up. Patients and methods The present study included 150 Egyptian patients with hepatitis C virus-related liver cirrhosis and ascites. The cases were divided into 100 patients with SBP and 50 patients with no SBP based on an elevated AF PMNL count of greater than or equal to 250 cells/mm3, ascitic samples were examined for PMNL count, culture, chemistry, and lactoferrin concentration in non-SBP patients and in SBP patients before and after systemic antibiotic treatment. Results AF lactoferrin concentration is significantly higher in SBP patients than in non-SBP patients with a cutoff value of 75.55 ng/ml, significantly higher in culture positive than in culture-negative SBP patients and its concentration is decreased significantly in SBP patients after systemic antibiotic therapy. Conclusion Elevated AF lactoferrin levels in cirrhotic patients are reliable for the diagnosis and follow up of SBP after systemic antibiotic therapy. The level of AF lactoferrin level is higher in resistant cases of SBP than the cases that respond to systemic antibiotic treatment.

Effect of oral administration of meloxicam prior to transport on inflammatory mediators and leukoctye function of cattle at feedlot arrival.

OBJECTIVE To investigate the effects of meloxicam administration before long-distance transport on inflammatory mediators and leukocyte function of cattle at feedlot arrival. ANIMALS 60 healthy yearling beef steers. PROCEDURES Single-source steers were assigned to a transported (n = 40) or nontransported (20) group. Then, half of the steers within each group were assigned to receive meloxicam (1 mg/kg, PO) or a lactose placebo (1 bolus/steer, PO). All steers were transported approximately 1,300 km overnight to a feedlot; however, the nontransported group was moved before treatment (meloxicam or placebo) administration and allowed a 17-day acclimation period, whereas the transported group was moved immediately after treatment administration on day -1. Blood samples for measurement of inflammatory mediators and leukocyte function were collected from all steers on days -1, 0, and 3. RESULTS For steers that received meloxicam, mean plasma meloxicam concentration for the transported group was significantly greater than that for the nontransported group on day 0. For steers that received the placebo, mean haptoglobin-matrix metalloproteinase-9 complex for the transported group was significantly greater than that for the nontransported group on day 0. Mean haptoglobin concentration, neutrophil L-selectin intensity, and polymorphonuclear leukocyte count for the transported group were significantly greater than those for the nontransported group. Mean substance P concentration for nontransported steers that received meloxicam was significantly lower than that for the other 3 treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated meloxicam administration to healthy steers immediately before long-distance transport did not significantly mitigate the effects of transport-induced stress on leukocyte function or inflammatory markers.

Current concepts and future strategies in the antimicrobial therapy of emerging Gram-positive spontaneous bacterial peritonitis.

Spontaneous bacterial peritonitis (SBP) is the most common infection in end-stage liver disease patients. SBP is defined as an ascitic fluid infection with a polymorphonuclear leucocyte count ≥ 250/mm3 without an evident intra-abdominal surgically treatable source. Several mechanisms contribute to SBP occurrence, including translocation of gut bacteria and their products, reduced intestinal motility provoking bacterial overgrowth, alteration of the gut’s barrier function and local immune responses. Historically, Gram-negative enteric bacteria have been the main causative agents of SBP, thereby guiding the empirical therapeutic choice. However, over the last decade, a worryingly increasing prevalence of Gram-positive and multi-drug resistant (MDR) SBP has been seen. Recently, the microbiological spectrum of SBP seems to have changed in Europe due to a high prevalence of Gram-positive bacteria (48%-62%). The overall proportion of MDR bacteria is up to 22%-73% of cases. Consequently, empirical therapy based on third-generation cephalosporins or amoxicillin/clavulanic acid, can no longer be considered the standard of care, as these drugs are associated with poor outcomes. The aim of this review is to describe, with an epidemiological focus, the evidence behind this rise in Gram-positive and MDR SBP from 2000 to present, and illustrate potential targeted therapeutic strategies. An appropriate treatment protocol should include daptomycin plus ceftaroline and meropenem, with prompt stepdown to a narrower spectrum when cultures and sensitivity data are available in order to reduce both cost and potential antibiotic resistance development.

Effects of intrauterine infusion of Escherichia coli lipopolysaccharide on uterine health, resolution of purulent vaginal discharge, and reproductive performance of lactating dairy cows

The objectives of the current experiment were to evaluate the effects of intrauterine infusion of Escherichia coli lipopolysaccharide (LPS) in cows diagnosed with purulent vaginal discharge (PVD) on intrauterine cell population, resolution of PVD, uterine health, and reproductive performance. Jersey cows (n = 3,084) were examined using the Metricheck device to diagnose PVD at 35 ± 6 d postpartum. Purulent vaginal discharge was defined as the presence of purulent (≥50% pus) discharge detectable in the vagina. Of the 310 cows positive for PVD, 267 cows were enrolled in the current experiment. To ensure proper timing of treatment and collection of samples, only 9 PVD-positive cows were treated per day. Selected cows were balanced at 35 ± 6 d postpartum for lactation number, body condition score, and milk yield and were randomly assigned to receive an intrauterine infusion of 20 mL of phosphate-buffered saline (PBS; control, n = 87), 20 mL of PBS with 150 µg LPS (LPS150, n = 91), or 20 mL of PBS with 300 µg of LPS (LPS300, n = 89). Uterine cytology was performed immediately before treatment and 1, 2, and 7 d after treatment to evaluate the effect of LPS treatment on intrauterine cell population. Cows were examined with the Metricheck device at 7 and 28 d after treatment to evaluate the effects of treatment on resolution of PVD. Reproductive status was recorded up to 200 d postpartum. Cows diagnosed with PVD had greater incidence of twinning, dystocia, retained placenta, and metritis after calving than cows without PVD. Count of polymorphonuclear leukocytes (PMNL) in uterine cytology 1, 2, and 7 d after intrauterine infusion was not statistically different among treatments. From d 0 to 1, however, PMNL count in uterine cytology of PBS cows increased by 5%, whereas the PMNL count in uterine cytology of LPS150 and LPS300 cows increased by 54 and 48%, respectively. Treatment did not affect the likelihood of cows being diagnosed with PVD 7 and 28 d after intrauterine infusion. Cows without PVD and LPS150 cows were more likely to be pregnant after the first postpartum AI than PBS cows. After the second postpartum AI, cows without PVD were more likely to be pregnant than PBS and LPS300 cows. Hazard of pregnancy up to 200 d postpartum was decreased for PBS and LPS300 cows compared with cows without PVD, and it tended to be decreased for LPS150 cows compared with cows without PVD. Intrauterine treatment with 150 µg of E. coli LPS of cows diagnosed with PVD improved likelihood of pregnancy after the first postpartum AI, but further research is needed to elucidate the mechanism by which LPS treatment improved fertility.