Fluid Tumor Necrosis Factor-alpha Research Articles

Salivary and crevicular fluid proinflammatory cytokines and advanced glycation end products in patients with different glycemic levels undergoing fixed orthodontic treatment.

To examine whether patients with different blood glycemic levels undergoing fixed orthodontic therapy demonstrate changes in the biochemical profiles of crevicular fluid and salivary advanced glycation end products (AGEs) and proinflammatory cytokine levels in comparison with nondiabetic healthy subjects. Prediabetic subjects, subjects with type 2 diabetes mellitus (T2DM), and subjects without a diabetes mellitus diagnosis undergoing fixed orthodontic therapy with MBT prescription brackets (0.022-inch brackets and 0.019 × 0.025-inch stainless steel archwires) were included in the study. The following clinical periodontal parameters were evaluated: (1) plaque score (PS), (2) probing depth (PD), (3) bleeding on probing (BOP), and (4) clinical attachment loss. Crevicular fluid and saliva specimens were collected during regular orthodontic visits. Salivary and crevicular fluid tumor necrosis factor alpha, interleukin-6, ghrelin, resistin, AGEs, and receptor activator of nuclear factor κΒ ligand were evaluated using a human magnetic Luminex multiplex assay. BOP scores were significantly higher among T2DM subjects (19.2%) than among nondiabetic subjects (11.2%) and prediabetic subjects (15.9%). Comparable values were demonstrated by all three study groups regarding PD scores and PSs. T2DM subjects demonstrated higher scores for gingival crevicular fluid (GCF) chemokines than nondiabetic and prediabetic subjects. A statistically significant difference was found in the levels of AGEs and resistin among the three study groups. The scores revealed for the levels of GCF resistin and AGEs versus periodontal BOP demonstrated a significant positive association by the Pearson correlation test. T2DM patients demonstrated significantly higher levels of GCF resistin and AGEs during fixed orthodontic therapy. Chronic hyperglycemic patients undergoing orthodontic therapy demonstrated a proinflammatory response.

Cerebrospinal fluid tumor necrosis factor-alpha (TNF-α) levels in children with cerebral malaria.

This prospective cross-sectional study evaluated the diagnostic and prognostic role of cerebrospinal fluid (CSF) tumor necrosis factor-alpha (TNF-α) in children with cerebral malaria (CM) and its role in the differentiation of CM from non-cerebral severe malaria. CSF TNF-α was measured using a human TNF-α enzyme-linked immunosorbent assay kit of 39 cases of CM and 19 cases of non-cerebral severe malaria. CSF TNF-α levels were significantly higher in CM (p < 0.001). Based on the receiver operating characteristics curve, a cutoff value of CSF TNF-α was 5.7 pg/ml for diagnosis of CM with sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of 87.2%, 94.7%, 97.1% and 78.3% respectively. The cutoff value of CSF TNF-α was 13.7 pg/ml for predicting adverse outcomes in CM with sensitivity, specificity, PPV and NPV of 100%, 96.8%, 88.9% and 100%, respectively. However, the cutoff value of CSF TNF-α was 4.96 pg/ml for predicting adverse outcomes in non-cerebral severe malaria with a sensitivity, specificity, PPV and NPV of 100%, 94.1%, 88.9% and 100% respectively. So, CSF TNF-α is an excellent biomarker and can be used as a diagnostic and prognostic tool. More studies are needed to establish CSF TNF-α as a predictor of neurological sequelae.

Effect of Taoren-Quyu decoction on endometriosis in rats

Purpose: To study the effect of traditional Chinese Medicine formula Taoren-Quyu decoction (TQD) on endometriosis.&#x0D; Method: Fifty female Wistar rats were randomly separated into five groups (10 rats/group): normal control, model (untreated) group, positive control (danazol), 200 mg/kg/day (low dose) or 400 mg/kg/day (high dose). All rats were prepared into endometriosis except for normal control rats. TDQ groups rats were orally administered of TQD for 5 weeks. After treatment, the rats were sacrificed by cervical dislocation. The number of total endometriotic lesions were counted. Serum levels of cancer antigen 125 (CA-125), interleukin 13 (IL-13), interleukin 18 (IL-18) and peritoneal fluid tumor necrosis factoralpha (TNF-α) were measured by ELISA kits.&#x0D; Result: Compared with control rats, TQD reduced the number of total endometriotic lesions significantly (12.7 ± 1.2, p &lt; 0.01), as well as serum levels of CA-125 (6.4 ± 1.2 U/mL), IL-18 (118.6 ± 7.4 pg/mL), IL13 (6.3 ± 0.8 pg/mL) and peritoneal fluid TNF-α (231.5 ± 11.7 pg/mL) (p &lt; 0.01).&#x0D; Conclusion: The results reveal that TQD exerts anti-endometriotic effect in rats by inhibiting inflammatory factors. Therefore, TQD has potentials for use in the treatment of endometriosis.

Prognostic role of follicular fluid tumor necrosis factor alpha in the risk of early ovarian hyperstimulation syndrome

BackgroundOvarian hyperstimulation syndrome (OHSS) is an iatrogenic condition characterized by capillary hyperpermeability which can be predicted by preovulatory ovarian responses such as number of follicles. A variety of cytokines are thought to be involved in pathophysiology of this syndrome.MethodsA prospective cohort study invloving sixty intracytoplasmic sperm injection (ICSI) patients. On the day of hCG injection, we explored the threshold of larger follicles ≥11 mm diameter with a count of ≥18 follicles for the high-risk moderate-to-severe OHSS and 13–18 follicles for the low-risk moderate-to-severe OHSS. Whereas larger follicles count of less than 13 were classified as normoresponders. Pooled follicular fluid (FF) samples of each patient were collected on the day of oocyte retrieval. Magnetic multiplex immunoassay was explored to measure the concentrations of some intrafollicular cytokines including: GM-CSF, INF-γ, TNF-α, IL-10, CXCL8/IL-8, IL-6, IL-5, IL-4, IL-2, and IL-1β. All sixty patients underwent controlled ovarian hyperstimulation (COH) with either GnRH agonist or antagonist protocols.ResultsIntrafollicular TNF-α concentration was significantly different (p < 0.05) in the high-risk moderate-to-severe OHSS patients compared to low-risk moderate-to-severe OHSS patients and normoresponders. TNF-α in FF had a negative correlation with the chance of high-risk moderate-to-severe OHSS. The differences in the risk of OHSS between patients who received GnRH agonist or antagonist were not significant (p > 0.05).ConclusionsIn accordance to the negative correlation of TNF-α and high risk of early OHSS, we did not expect TNF-α to play a role in increasing vascular permeability in ovarian tissues. In addition, the risk of early moderate-to-severe OHSS was not affected by different GnRH superovulation protocols.

Tumor necrosis factor alpha and interleukin-1 beta levels in cerebrospinal fluid examination for the diagnosis of ventriculoperitoneal shunt-related ventriculitis.

Ventriculitis is known to develop after chronic inflammation and bacterial invasion of the ventricular surface with a recurrence of shunt infections. The aim of this study is to evaluate the diagnostic value of elevation in cerebrospinal fluid (CSF) interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) together with CSF culture and laboratory test results in the diagnosis of ventriculoperitoneal (VP) shunt-related ventriculitis, which is known to be more problematic than conventional shunt infection. The study included a total of 34 patients with a VP shunt due to hydrocephalus, who presented with a headache, fever, and shunt infection at the Emergency Department and had a pre-diagnosis of ventriculitis. Nineteen patients were diagnosed with shunt-related infection or ventriculitis using the CSF obtained from the shunt pump. The IL-1β and TNF-α levels from the CSF samples of all patients were measured using the Micro ELISA immunoassay method. CSF direct microscopic observation revealed that the mean cell count, IL-1β level, CRP level, and blood leukocyte level were higher in patients with ventriculitis compared to those diagnosed with shunt infection (p= 0.02, p= 0.009, p= 0.004, and p = 0.009, respectively). The probability of predicting positive culture outcome was 92.7% with 90.9% sensitivity and 82.6% specificity when IL-1β values exceeded 4.0pg/ml. TNF-α values did not show a significant, reliable pattern compared to IL-1β. IL-1β is a reliable parameter which shall be used in the diagnosis of ventriculitis by predicting positive culture outcome with high sensitivity and specificity.

Association between metabolic syndrome and gingival inflammation in obese children.

Our previous work showed a positive association between metabolic syndrome (MetS) and gingival crevicular fluid (GCF) tumour necrosis factor-alpha (TNF-α) in a sample of obese and non-obese children. However, whether this association persists among obese children is unknown. We aim to investigate the extent to which MetS is associated with GCF TNF-α level among obese children. We performed a cross-sectional analysis using data from visit 1 of the QUebec Adipose and Lifestyle InvesTigation in Youth cohort. A total of 219 obese children aged 8-10years, for whom data were available for both MetS and TNF-α, were included in our analysis. The independent variable, MetS, was defined according to the International Diabetes Federation recommendations. GCF samples were collected from the gingival sulcus using a paper strip, and the concentration of TNF-α was determined by enzyme-linked immunosorbent assay. Analyses included descriptive statistics and sex-specific linear regression analyses adjusting for potential confounders. In this sample comprising only obese children, 24 (10.9%) had MetS. Among obese boys, those with MetS had 44.9% higher GCF TNF-α (95% confidence interval: 16.5%-73.3%) compared to those without MetS. No such association was detected in obese girls. MetS was positively associated with GCF TNF-α concentration in obese boys. These results suggest that obese boys with MetS may have a worse gingival health profile compared to their obese counterpart without MetS.

Experimental study of cerebrospinal fluid tumor necrosis factor-alpha release in penicillin- and cephalosporin-resistant pneumococcal meningitis treated with different antibiotic schedules.

To measure the inflammatory response in terms of tumor necrosis factor-alpha (TNF-α) levels in cerebrospinal fluid (CSF), using bacteriolytic versus nonbacteriolytic antibiotic therapy and adjunctive treatment with dexamethasone in an experimental rabbit model of pneumococcal meningitis. In a rabbit model of pneumococcal meningitis, we tested CSF TNF-α levels in several samples from rabbits infected with the HUB 2349 strain and treated with ceftriaxone 100mg/kg/d, ceftriaxone plus vancomycin 30mg/kg/d, or daptomycin at 15mg/kg or 25mg/kg. Daptomycin schedules were compared with the same doses in combination with dexamethasone at 0.125mg/kg every 12hours over a 26-hour period. The ceftriaxone group had the highest levels of TNF-α. TNF-α levels were significantly higher after ceftriaxone administration than in both daptomycin groups. The high-dose daptomycin group presented the lowest inflammatory levels in CSF samples. Adjunctive treatment with dexamethasone in this group modulated the inflammatory response, bringing down CSF TNF-α levels. CSF TNF-α levels were significantly lower in rabbits treated with daptomycin than in rabbits treated with ceftriaxone. Daptomycin avoided the inflammatory peak after administration observed in ceftriaxone-treated rabbits. The use of daptomycin plus dexamethasone achieved a significantly larger reduction in CSF TNF-α levels.

Significance of cerebrospinal fluid matrix metalloproteinase-9, interleukin-6, and tumor necrosis factor-alpha protein in children with viral encephalitis

Objective To explore the significance of changes of matrix metalloproteinase-9(MMP-9), interleukin-6(IL-6), and tumor necrosis factor-alpha(TNF-α) protein level in the cerebrospinal fluid (CSF) of children with viral encephalitis(VE). Methods The concentration of neuron-specific enolase(NSE), structural proteins 100B (S100B), and MMP-9, IL-6, TNF-α in the CSF of VE children were detected by an enzyme linked immunosorbent assay, and the correlations of them were analyzed. Results NSE, S100B, MMP-9, IL-6, TNF-α protein expression could be found significantly higher than those in the control group, and there were significant differences according to statistics expression trends(all P<0.05). The NSE protein expression was significantly positive related with S100B in the VE group (r=0.467, P=0.009), and the concentration was markedly negative related with the duration of viral encephalitis (r=-0.472, P=0.008). MMP-9, IL-6 protein expression were significantly positive related with NSE, S100B respectively (r=0.698, P=0.00; r=0.559, P=0.00; r=0.812, P=0.00; r=0.664, P=0.00). TNF-α protein expression was positive related with CSF S100B(r=0.363, P=0.049), but there was no correlation between TNF-α and NSE(r=0.245, P=0.193). Conclusions The neurons and the neuroglial cells are damaged in the viral encephalitis children.MMP-9, IL-6, TNF-α protein may participate in the pathological damage process of nerve cells in VE children in different degrees. Key words: Viral encephalitis; Structural proteins 100B; Matrix metalloproteinase-9; Interleukin-6; Tumor necrosis factor-alpha; Child

Circulating undercarboxylated osteocalcin and gingival crevicular fluid tumour necrosis factor‐α in children

Osteocalcin, a protein secreted by osteoblasts during bone formation, is negatively associated with adult periodontal disease. Little is known about this association in children. To examine the extent to which plasma undercarboxylated osteocalcin (ucOC) is associated with gingival crevicular fluid tumour necrosis factor-alpha (GCF TNF-α) - a potential marker of gingival inflammation - in children. We used data from the Quebec Adipose and Lifestyle InvesTigation in Youth cohort, an ongoing longitudinal study on the natural history of obesity among Caucasian children with a family history of obesity in Quebec, Canada. This cross-sectional analysis from the baseline visit includes 120 children aged 8-10years. Plasma ucOC and GCF TNF-α levels were determined by enzyme-linked immunosorbent assay. Linear regression analyses, adjusting for age, gender, family income, sexual maturity stage, daily physical activity, obesity, and fasting glucose were conducted, with TNF-α level as the dependent variable. A 1-ng/ml increase in ucOC was associated with a 0.96% decrease (95% confidence interval (CI): -1.69, -0.23) in GCF TNF-α level. A negative association between a marker of bone formation and a marker of gingival inflammation was observed as early as childhood among Caucasian children with a family history of obesity.

Metabolic syndrome and gingival inflammation in Caucasian children with a family history of obesity

To investigate whether metabolic syndrome (MetS) and its components are associated with gingival inflammation in children. This is a cross-sectional analysis from the baseline visit of the QUebec Adipose and Lifestyle InvesTigation in Youth cohort, an ongoing longitudinal study investigating the natural history of obesity in children of Quebec, Canada. The analytic sample includes 448 children aged 8-10 years, 39% of whom were overweight or obese. MetS was defined according to the International Diabetes Federation recommendations. Gingival inflammation was defined by the level of gingival crevicular fluid (GCF) tumour necrosis factor alpha (TNF-α) and the extent of gingival bleeding. Sex-specific linear regression analyses estimated the associations between MetS and gingival inflammation, adjusting for potential confounders. Twenty-five children had MetS. Boys with MetS compared to those without, had a 49.5% (p-value = 0.001) higher GCF TNF-α level and 13.7% (p-value = 0.033) more sites with gingival bleeding. Moreover, for three of the five components of MetS - waist circumference, fasting plasma triglycerides, systolic blood pressure - an increase was associated with increased GCF TNF-α level in boys. No such findings were seen in girls. An association between MetS and gingival inflammation was observed as early as in childhood, and may differ by sex.

The Values of Serum and Ascetic Fluid Tumor Necrosis Factor-Alpha (TNF- alpha), TNF-Receptor 1 and C-Reactive Protein (CRP) in Patients with Cirrhotic Ascites

Spontaneous bacterial peritonitis (SBP) is defined as an infection of initially sterile ascitic fluid (AF) without a detectable, surgically treatable source of infection. It is a frequent and severe complication of cirrhotic ascites. Because of the high morbidity and mortality of SBP, the rapid and accurate diagnosis is required. Aim: The present study aimed to measure the levels of tumor necrosis factor-alpha (TNF-alpha), tumor necrosis factor receptor (TNF-r p55) and C-reactive protein (CRP) in both ascetic fluid and serum of patients with sterile and infected cirrhotic ascites to show their diagnostic values as compared to ascitic fluid culture and polymorphnuclear leukocyte (PMN) count. Patients and methods: TNF-alpha, TNF-r p55 and CRP were measured in both ascetic fluid and serum of 20 patients with spontaneous bacterial peritonitis (SBP), in addition to 22 patients with sterile cirrhotic ascites. Results: The results of clinical examination showed a significant difference as regard abdominal pain, fever jaundice, upper GIT bleedings, encephalopathy and Sclerotherapy among both groups. The serum levels of CRP and TNF-alpha were significantly higher in patients with SBP as comparing to patients have sterile ascites, but TNF-r p55 serum level showed no significant difference. On evaluation of ascetic fluid parameters, total leucocytic count (TLC), plymorphnuclear (PMN) count , CRP, TNF-alpha, TNF-r p55 are significantly higher in SBP patient group than group of sterile ascites. Sensitivity and specificity of ascitic fluid PMN (cut-off value >250 cells/ mm3) in discriminating infected ascites from sterile ascites were 70% and 86.4%, respectively. Sensitivity and specificity of ascitic fluid CRP (cut-off value >1.0 mg/dL) in discriminating infected ascites from sterile ascites were 85% and 72.7%, respectively. Sensitivity and specificity of ascitic fluid TNF-alpha (cut off value >12 pg/ml) in discriminating infected ascites from sterile ascites were 80% and 63.6%, respectively. Sensitivity and specificity of TNF-r p55 (cut-off value >6.2 pg/ml) in discriminating infected ascites from sterile ascites were 75% and 68.2%, respectively. Conclusion: We concluded that, the elevated serum and ascetic fluid levels of CRP, TNF-alpha and TNF-r may suggest their role in the pathogenesis of ascetic fluid infection and their higher sensitivity and specificity make them to be good discriminators in ascetic fluid infection (especially a cheap and easy ascitic fluid CRP levels). Thus may help in rapid diagnosis and early start empirical antibiotic therapy without waiting the culture results.

The Values of Serum and Ascetic Fluid Tumor Necrosis Factor-Alpha (TNF- alpha), TNF-Receptor 1 and C-Reactive Protein (CRP) in Patients with Cirrhotic Ascites

Spontaneous bacterial peritonitis (SBP) is defined as an infection of initially sterile ascitic fluid (AF) without a detectable, surgically treatable source of infection. It is a frequent and severe complication of cirrhotic ascites. Because of the high morbidity and mortality of SBP, the rapid and accurate diagnosis is required. Aim: The present study aimed to measure the levels of tumor necrosis factor-alpha (TNF-alpha), tumor necrosis factor receptor (TNF-r p55) and C-reactive protein (CRP) in both ascetic fluid and serum of patients with sterile and infected cirrhotic ascites to show their diagnostic values as compared to ascitic fluid culture and polymorphnuclear leukocyte (PMN) count. Patients and methods: TNF-alpha, TNF-r p55 and CRP were measured in both ascetic fluid and serum of 20 patients with spontaneous bacterial peritonitis (SBP), in addition to 22 patients with sterile cirrhotic ascites. Results: The results of clinical examination showed a significant difference as regard abdominal pain, fever jaundice, upper GIT bleedings, encephalopathy and Sclerotherapy among both groups. The serum levels of CRP and TNF-alpha were significantly higher in patients with SBP as comparing to patients have sterile ascites, but TNF-r p55 serum level showed no significant difference. On evaluation of ascetic fluid parameters, total leucocytic count (TLC), plymorphnuclear (PMN) count , CRP, TNF-alpha, TNF-r p55 are significantly higher in SBP patient group than group of sterile ascites. Sensitivity and specificity of ascitic fluid PMN (cut-off value >250 cells/ mm3) in discriminating infected ascites from sterile ascites were 70% and 86.4%, respectively. Sensitivity and specificity of ascitic fluid CRP (cut-off value >1.0 mg/dL) in discriminating infected ascites from sterile ascites were 85% and 72.7%, respectively. Sensitivity and specificity of ascitic fluid TNF-alpha (cut off value >12 pg/ml) in discriminating infected ascites from sterile ascites were 80% and 63.6%, respectively. Sensitivity and specificity of TNF-r p55 (cut-off value >6.2 pg/ml) in discriminating infected ascites from sterile ascites were 75% and 68.2%, respectively. Conclusion: We concluded that, the elevated serum and ascetic fluid levels of CRP, TNF-alpha and TNF-r may suggest their role in the pathogenesis of ascetic fluid infection and their higher sensitivity and specificity make them to be good discriminators in ascetic fluid infection (especially a cheap and easy ascitic fluid CRP levels). Thus may help in rapid diagnosis and early start empirical antibiotic therapy without waiting the culture results.

Investigation of changes of the level of IL-1 and TNF-α on the pathogenesis of patients with knee osteoarthritis

Objective To explore the relationships between the levels of synovial fluid and serum interleukin-1(IL-1)and tumor necrosis factor alpha(TNF-α)and severity of knee osteoarthritis (OA). Methods 50 patients with unilateral OA(18 cases in early stage,15 cases in middle stage,and 17 cases in terminal stage)were included in the study.15 healthy volunteers were examined as the control group.Synovial fluid and serum of all patients and volunteers were collected for measurement of IL-1 and TNF-α. Results The IL-1 and TNF-α levels of synovial fluids and serum in OA group were significantly higher than those of the control group(P <0.01).With the progress of OA,the IL-1 and TNF-α levels were increased significantly(P <0.01).IL-1 level in synovial fluid and serum of OA group was correlated positively with TNF-α level(P <0.01). Conclusion IL-1 and TNF-α were related to OA patients with immune inflammation,and their levels could reflect changes in disease activity.The increasing IL-1 and TNF-α levels in synovial fluids and serum were positively correlated with the progress of OA.Therefore,combined detection of these two indexes could be useful for determining the pathogenetic condition of KOA. Key words: Osteoarthritis; Synovial fluid; Interleukin-1 ; Tumor necrosis factor alpha

Adiposity and gingival crevicular fluid tumour necrosis factor‐α levels in children

To investigate whether adiposity is associated with gingival crevicular fluid (GCF) tumour necrosis factor-alpha (TNF-alpha) levels in children. We also examined whether this relationship is mediated through plasma fasting insulin. This preliminary study used cross-sectional data from the baseline-visit of the Quebec Adipose and Lifestyle InvesTigation in Youth cohort, which is an ongoing longitudinal study investigating the natural history of obesity in Quebec children. Study participants (76 girls and 102 boys) include children aged 8-10 years and their families, living in the Montreal and Quebec City areas. TNF-alpha level was measured in pooled samples (N=4) for each child by enzyme-linked immunosorbant assay. Height and weight were measured. Body mass index (BMI) was calculated as weight/height(2) (kg/m(2)). Sex/age-specific BMI was categorized into normal (<85th percentile), overweight (85th-95th percentile) and obese (>or=95th percentile) defined by the 2000 US-CDC growth charts. Insulin resistance was measured using fasting plasma insulin in children. Data analysis involved descriptive and multiple linear regression analyses. Our results suggest that obesity in boys was associated with a 37% increase of GCF-TNF-alpha level. However, when accounting for insulin resistance this association was reduced and disappeared while the model's goodness of fit improved. These findings provide support for the link between adiposity in children and GCF-TNF-alpha level, which appears to be mediated by insulin resistance.

The Endothelin-Converting Enzyme-1/Endothelin-1 Pathway Plays a Critical Role in Inflammation-Associated Premature Delivery in a Mouse Model

Premature delivery occurs in 12% of all births and accounts for nearly half of long-term morbidity. Current therapeutic approaches to preterm delivery are ineffective and present serious risks to both mother and fetus. The single most common cause of preterm birth is infection. Previous in vitro investigations have shown that endothelin-1 (ET-1) is induced by inflammatory cytokines and that it increases myometrial smooth muscle tone. Furthermore, we have previously shown that both the endothelin-converting enzyme-1 (ECE-1) inhibitor, phosphoramidon, as well as a novel ET-1 receptor A antagonist synthesized by our group, control premature delivery in a mouse model of inflammation-associated preterm delivery. In the current work, we show that levels of both ET-1 and ECE-1 are increased in gestational tissues in E16.5 mice induced to deliver prematurely after lipopolysaccharide administration. We also show that premature delivery is controlled by treatment with the selective endothelin receptor A antagonist BQ-123 in a dose-dependent manner. Finally, we show here for the first time that premature delivery can be controlled using RNA silencing, by hydrodynamic transfection of E15 mice with ECE-1 RNAi. Taken together, these data support a critical role for the ECE-1/ET-1 system in inflammation-associated premature delivery. The ability to control premature delivery by antagonizing or silencing the ECE-1/ET-1 system offers a novel approach to an unmet clinical need.

Bacterial DNA in patients with cirrhosis and noninfected ascites mimics the soluble immune response established in patients with spontaneous bacterial peritonitis

Bacterial infections and severity of associated inflammatory reaction influence prognosis in patients with advanced cirrhosis. We compared the innate immune response to bacterial DNA (bactDNA) translocation with that caused by viable bacteria translocation in patients with spontaneous bacterial peritonitis and the relationship between the cytokine response and serum levels of bactDNA. The bactDNA translocation was investigated in 226 patients with cirrhosis and noninfected ascites, 22 patients with spontaneous bacterial peritonitis, and 10 patients with ascites receiving continuous norfloxacin. Serum and ascitic fluid tumor necrosis factor alpha, interferon-gamma, interleukin-12, and nitric oxide metabolites were measured via enzyme-linked immunosorbent assay. Bacterial genomic identifications were made via amplification and sequencing of the 16S ribosomal RNA gene and digital quantization with DNA Lab-on-chips. The bactDNA was present in 77 noninfected patients (34%) and in all cases of spontaneous bacterial peritonitis, even in those with culture-negative ascitic fluid. No patient receiving norfloxacin showed bactDNA translocation. Levels of all cytokines were similar in patients with bactDNA translocation or spontaneous bacterial peritonitis and significantly higher than in patients without bactDNA or in those receiving norfloxacin. Serum bactDNA concentration paralleled levels of all cytokines and nitric oxide in a series of patients with bactDNA translocation or spontaneous bacterial peritonitis followed during 72 hours. Antibiotic treatment in the series of patients with spontaneous bacterial peritonitis did not abrogate bactDNA translocation in the short term. bactDNA translocation-associated cytokine response is indistinguishable from that in patients with spontaneous bacterial peritonitis and is dependent on bactDNA concentration. Norfloxacin abrogates bactDNA translocation and cytokine response.

Changes in Cytokine Levels and CD4+/CD8+ T Cells Ratio in Draining Lymph Node of Burn Wound

The aim of this study was to determine the changes of cytokine levels in draining lymph fluid and the changes of CD4+/CD8+ T-cells ratio in draining lymph node of burn wound. Male Wistar rats were subjected to unilateral hind limb burn (burn limbs group) and contralateral hind limb without burn (control limbs group). On hours 6, 24, and 72 after burn, rats were killed; lymph fluid in the efferent lymph trunk of the common iliac lymph nodes (CILN) were collected; and lymph fluid Interferon-gamma (IFN-gamma), interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-alpha) concentrations were measured by enzyme-linked immunosorbent assay. The CD4+/CD8+ T cells ratio of CILN was submitted to flow cytometry. The results showed TNF-alpha concentrations were significantly greater in the burn limbs group when compared with the control limbs group (P < .05). The IFN-gamma/IL-4 ratio was significantly lower (P < .05). The CD4+/CD8+ T-cell ratio was significantly lower on postburn hours 72 (P < .05). This study provides evidence that the burn wound can increase TNF-alpha levels and decrease IFN-gamma/ IL-4 ratio in the draining lymph fluid and decrease CD4+/CD8+ T cells ratio in the draining lymph node.

Lymph invasion route of bacteria and endotoxin and CD4+/CD8+ T cells ratio changes of draining lymph node of burn infection wound

The aim of this study was to determine the lymphatic invasion route of bacteria and endotoxin of burn wounds infected by Pseudomonas aeruginosa and moreover, the effect of P. aeruginosa infection of the burn wound on the draining lymph node and lymph fluid. Male Wistar rats were subjected to unilateral hind limb burn + wound infected by P. aeruginosa (infection limbs group) and contralateral hind limb burn alone (burn limbs group). On hours 6, 24 and 72 after infection, rats were killed, the common iliac lymph nodes (CILN) was collected for the culture of P. aeruginosa. Lymph fluid in the efferent lymph trunk of CILN was collected for the measurement of endotoxin by the Limulus Amebocyte Lysate test. Lymph fluid tumor necrosis factor-alpha (TNF-α) concentration was measured by enzyme-linked immunosorbent assay (ELISA). The CD4+/CD8+ T cells ratio of CILN was subjected to flow cytometry analysis. The results showed bacteria invasion incidence, endotoxin and TNF-α concentrations were significantly higher in the infection limb group when compared to the burn limb group ( P < 0.01). The CD4+/CD8+ T cell ratio was significantly lower on post-burn wound infection hours 72 ( P < 0.05). This study provides evidence that bacteria and endotoxin of burn wound infected by P. aeruginosa invade draining lymph node and lymph fluid. P. aeruginosa infection of the burn wound can increase TNF-α level of the draining lymph fluid and decrease CD4+/CD8+ T cells ratio of the draining lymph node.

Disruption of Interleukin-18, but Not Interleukin-1, Increases Vulnerability to Preterm Delivery and Fetal Mortality after Intrauterine Inflammation

Preterm birth is a major contributor of adverse perinatal outcome. Clinical data suggest that an inflammatory response is important in the process leading to preterm labor. By using a recently introduced mouse model of localized intrauterine lipopolysaccharide-induced inflammation, the effect of interleukin (IL)-18 gene disruption and/or IL-18 neutralization as well as combined IL-1alpha/beta gene disruption on inflammation-induced fetal loss was investigated. The frequency of preterm fetal loss was significantly higher in IL-18 knockout mice (58.9%) and in mice administered IL-18-binding protein (59.7%) compared to wild-type controls (34.7%). The rate of fetal loss was not affected by IL-1alpha/beta gene deficiency (38.7%). Decreased IL-18 protein expression combined with elevated IL-12 protein expression in uterine tissue of IL-18 knockout mice and IL-18-binding protein-treated animals was noticed. These data demonstrate that preterm pregnancy loss in response to intrauterine inflammation was enhanced by disruption of the IL-18 gene and/or IL-18 neutralization, events that may relate to exaggerated Th1 responses because of an increased IL-12/IL-18 ratio.

Extracellular 70-kd heat shock protein in mid-trimester amniotic fluid and its effect on cytokine production by ex vivo–cultured amniotic fluid cells

The 70-kd heat shock protein is released from cells in response to stress and functions as a regulator of innate immunity. We hypothesized that 70-kd heat shock protein in mid-trimester amniotic fluid might regulate local immune system activation. Amniotic fluid that was obtained from 200 women who underwent amniocentesis at 15 to 19 weeks of gestation was tested by enzyme-linked immunosorbent assay for 70-kd heat shock protein, tumor necrosis factor-alpha, and interleukin-1beta and -6. The amniotic fluid cellular fraction also was evaluated for Mycoplasma hominis by gene amplification. Whole amniotic fluids were incubated ex vivo in medium alone or medium that contained peptidoglycan, a TLR2 ligand, or lipopolysaccharide, a TLR4 ligand. After 24 hours, the supernatants were collected and assayed for 70-kd heat shock protein. The influence of exogenous 70-kd heat shock protein on tumor necrosis factor-alpha and interleukin-1beta and -6 production by whole amniotic fluid was assessed similarly. The 70-kd heat shock protein was detected in all amniotic fluids with a median (range) of 11.5 ng/mL (1.2-76.7). The intra-amniotic 70-kd heat shock protein concentration was correlated positively only with amniotic fluid tumor necrosis factor-alpha levels (P = .0002). Detection of M hominis was associated with an increased 70-kd heat shock protein concentration (median, 17.2 ng/mL; P = .01). The addition of peptidoglycan resulted in a stimulation of 70-kd heat shock protein production, and exogenous 70-kd heat shock protein stimulated the release of tumor necrosis factor-alpha by amniotic fluid cells. The 70-kd heat shock protein is released from cells in mid-trimester amniotic fluid as a consequence of TLR2 stimulation and potentiates tumor necrosis factor-alpha production.