Lower Concentrations Of IL-1β Research Articles

Markers of chronic low-grade inflammation and serum cytokine levels in patients with type 1 diabetes: associations with time in ranges and glucose variability

BACKGROUND: Increased glucose variability is recognized as a risk factor for vascular diabetic complications. It is assumed that deteriorating effect of GV on blood vessels can be realized through the activation of inflammatory signaling pathways.AIM: to determine associations of low-grade inflammation markers and serum cytokines with time in ranges and GV parameters derived from continuous glucose monitoring (CGM) in patients with type 1 diabetes (T1D).MATERIALS AND METHODS: In 470 adult patients with T1D, high-sensitivity C-reactive protein (hsCRP) and fibrinogen was measured, neutrophil-lymphocyte ratio (NLR) and the Systemic Immune-inflammation Index (SII) were calculated. In a sample of 130 patients and 20 healthy individuals (control), serum concentrations of interleukins (IL-1β, IL-4, IL-6, sIL-6Rα, IL-19, IL-20, IL-22, IL-26, IL-27, IL-28A, IL-29, IL-32, IL-34, IL-35) were assessed by multiplex analysis. Time in the ranges and GV parameters: Coefficient of Variability (CV), Mean Amplitude of Glycemic Excursions (MAGE), and Mean Absolute Glucose rate of changes (MAG) were derived from CGM data.RESULTS: Patients with Time In Range (TIR) <70% had higher concentrations of hs-CRP and fibrinogen, higher SII values, and demonstrated a trend toward higher TIR compared with those with TIR ≥70% (p=0.018, p=0.026, p=0.037, p=0.101, respectively). Patients with T1D, when compared to control, demonstrated increased concentrations of IL-1β (p<0.0001), IL-6 (p<0.0001), decreased levels of IL-4 (p=0.002), and a tendency to decrease IL-22 and IL-29 (p=0.1). Patients with TIR>70% had higher levels of IL-4 (p=0.02) as well as lower concentrations of IL-1β (p=0.0003) and IL-6 (p=0.007) than patients with TIR≤70%. In a multivariate stepwise regression analysis including clinical data and CGM parameters as independent variables, body mass index was positive predictor of hsCRP and fibrinogen levels, TIR was negatively associated with IL-20 and IL-34, time above range was associated positively with IL-1β, MAGE showed positive association with SII, IL-26 and IL28A, while MAG was positively associated with IL-29.CONCLUSION: T1D patients with non-target TIR (<70%) have higher levels of low-intensity inflammatory markers and serum pro-inflammatory cytokines than patients with TIR>70%. Both hyperglycemia and increased GV are associated with intensity of low-grade inflammation in T1D.

Qingre Huoxue Decoction regulates macrophage polarisation to attenuate atherosclerosis through the inhibition of NF-κB signalling-mediated inflammation

Ethnopharmacological relevanceAtherosclerosis (AS) is a common pathogenesis of cardiovascular diseases. Qingre Huoxue Decoction (QRHX) is an herbal formula used for the prevention and treatment of AS. However, the potential mechanism of QRHX is not clear. Aim of the studyIn our study, RNA sequencing combined with preclinical models were used to analyse the effect and mechanism of QRHX for the treatment of AS. Materials and methodsFor in vivo studies, ApoE−/− mice were fed with a high-fat diet to induce AS. We measured weight, blood lipid, inflammatory cytokines, lipid deposition, plaque, and the M1/M2 macrophage. For in vitro studies, RAW264.7 were induced by lipopolysaccharides and treated with different concentrations of QRHX. We focusd on the relationship between QRHX, the NF-κB pathway, and macrophage polarisation, and performed simultaneous RNA sequencing both in vivo and in vitro. ResultsIn vivo, QRHX decreased weight, improved blood lipid, relieved the degree of lipid deposition, reduced plaque area, decreased the levels of inflammatory cytokines (MCP-1, NLRP3, and TNFα), down-regulated the expression of iNOS, and up-regulated the expression of Arg-1. In vitro, QRHX down-regulated M1 markers, iNOS and CCR7, with lower concentrations of IL-1β; furthermore, QRHX up-regulated M2 markers, Arg-1, CD163, Ym-1, and Fizz-1, with higher concentrations of IL-4 and IL-10. RNA sequencing of both samples in vivo and in vitro suggested that NF-κB was the target pathway of QRHX to regulate macrophage polarisation; this result was validated at the gene and protein levels. ConclusionsQRHX induced M2 polarisation, reduced an inflammatory response, and played a role in stabilising plaque by mediating the NF-κB signalling pathway.

Effects of Rhamnolipids on Growth Performance, Immune Function, and Cecal Microflora in Linnan Yellow Broilers Challenged with Lipopolysaccharides.

This present study aimed to investigate the effects of rhamnolipids (RLS) on the growth performance, intestinal morphology, immune function, short-chain fatty acid content, and microflora community in broiler chickens challenged with lipopolysaccharides (LPS). A total of 450 broiler chickens were randomly allocated into three groups: basal diet with no supplement (NCO), basal diet with bacitracin (ANT), and basal diet with rhamnolipids (RLS). After 56 d of feeding, 20 healthy broilers were selected from each group, with half being intraperitoneally injected with lipopolysaccharides (LPS) and the other half with normal saline. Treatments with LPS were labelled LPS-NCO, LPS-ANT, and LPS-RLS, whereas treatments with normal saline were labelled NS-NCO, NS-ANT, and NS-RLS. LPS-challenged birds had lower jejunal villus height and higher crypt depth than unchallenged birds. LPS-RLS broilers had increased jejunal villus height and villus height/crypt depth ratio (V/C) but lower crypt depth than LPS-NCO. Dietary supplementation with RLS reduced the LPS-induced immunological stress. Compared with LPS-NCO, birds in LPS-RLS had lower concentrations of IL-1β, IL-6, and TNF-α. In LPS-challenged broilers, RLS and ANT increased the concentrations of IgA, IgM, and IgY compared with LPS-NCO. In LPS treatments, RLS enhanced the contents of acetic acid, butyrate, isobutyric acid, isovalerate, and valerate more than LPS-NCO birds. High-throughput sequencing indicated that RLS supplementation led to changes in the cecal microbial community of broilers. At the species level, Clostridium-sp-Marseille-p3244 was more abundant in NS-RLS than in NS-NCO broilers. In summary, RLS improved the growth performance and relative abundance of cecal microbiota and reduced the LPS-induced immunological stress in broiler chickens.

Low concentration IL-1β promotes islet amyloid formation by increasing hIAPP release from humanised mouse islets in vitro.

Aggregation of the beta cell secretory product human islet amyloid polypeptide (hIAPP) results in islet amyloid deposition, a pathological feature of type 2 diabetes. Amyloid formation is associated with increased levels of islet IL-1β as well as beta cell dysfunction and death, but the mechanisms that promote amyloid deposition in situ remain unclear. We hypothesised that physiologically relevant concentrations of IL-1β stimulate beta cell islet amyloid polypeptide (IAPP) release and promote amyloid formation. We used a humanised mouse model of endogenous beta cell hIAPP expression to examine whether low (pg/ml) concentrations of IL-1β promote islet amyloid formation in vitro. Amyloid-forming islets were cultured for 48h in the presence or absence of IL-1β with or without an IL-1β neutralising antibody. Islet morphology was assessed by immunohistochemistry and islet mRNA expression, hormone content and release were also quantified. Cell-free thioflavin T assays were used to monitor hIAPP aggregation kinetics in the presence and absence of IL-1β. Treatment with a low concentration of IL-1β (4pg/ml) for 48h increased islet amyloid prevalence (93.52 ± 3.89% vs 43.83 ± 9.67% amyloid-containing islets) and amyloid severity (4.45 ± 0.82% vs 2.16 ± 0.50% amyloid area/islet area) in hIAPP-expressing mouse islets in vitro. This effect of IL-1β was reduced when hIAPP-expressing islets were co-treated with an IL-1β neutralising antibody. Cell-free hIAPP aggregation assays showed no effect of IL-1β on hIAPP aggregation in vitro. Low concentration IL-1β did not increase markers of the unfolded protein response (Atf4, Ddit3) or alter proIAPP processing enzyme gene expression (Pcsk1, Pcsk2, Cpe) in hIAPP-expressing islets. However, release of IAPP and insulin were increased over 48h in IL-1β-treated vs control islets (IAPP 0.409 ± 0.082 vs 0.165 ± 0.051pmol/5 islets; insulin 87.5 ± 8.81 vs 48.3 ± 17.3pmol/5 islets), and this effect was blocked by co-treatment with IL-1β neutralising antibody. Under amyloidogenic conditions, physiologically relevant levels of IL-1β promote islet amyloid formation by increasing beta cell release of IAPP. Neutralisation of this effect of IL-1β may decrease the deleterious effects of islet amyloid formation on beta cell function and survival.

Biomarker-guided antibiotic stewardship in suspected ventilator-associated pneumonia (VAPrapid2): a randomised controlled trial and process evaluation

SummaryBackgroundVentilator-associated pneumonia is the most common intensive care unit (ICU)-acquired infection, yet accurate diagnosis remains difficult, leading to overuse of antibiotics. Low concentrations of IL-1β and IL-8 in bronchoalveolar lavage fluid have been validated as effective markers for exclusion of ventilator-associated pneumonia. The VAPrapid2 trial aimed to determine whether measurement of bronchoalveolar lavage fluid IL-1β and IL-8 could effectively and safely improve antibiotic stewardship in patients with clinically suspected ventilator-associated pneumonia.MethodsVAPrapid2 was a multicentre, randomised controlled trial in patients admitted to 24 ICUs from 17 National Health Service hospital trusts across England, Scotland, and Northern Ireland. Patients were screened for eligibility and included if they were 18 years or older, intubated and mechanically ventilated for at least 48 h, and had suspected ventilator-associated pneumonia. Patients were randomly assigned (1:1) to biomarker-guided recommendation on antibiotics (intervention group) or routine use of antibiotics (control group) using a web-based randomisation service hosted by Newcastle Clinical Trials Unit. Patients were randomised using randomly permuted blocks of size four and six and stratified by site, with allocation concealment. Clinicians were masked to patient assignment for an initial period until biomarker results were reported. Bronchoalveolar lavage was done in all patients, with concentrations of IL-1β and IL-8 rapidly determined in bronchoalveolar lavage fluid from patients randomised to the biomarker-based antibiotic recommendation group. If concentrations were below a previously validated cutoff, clinicians were advised that ventilator-associated pneumonia was unlikely and to consider discontinuing antibiotics. Patients in the routine use of antibiotics group received antibiotics according to usual practice at sites. Microbiology was done on bronchoalveolar lavage fluid from all patients and ventilator-associated pneumonia was confirmed by at least 104 colony forming units per mL of bronchoalveolar lavage fluid. The primary outcome was the distribution of antibiotic-free days in the 7 days following bronchoalveolar lavage. Data were analysed on an intention-to-treat basis, with an additional per-protocol analysis that excluded patients randomly assigned to the intervention group who defaulted to routine use of antibiotics because of failure to return an adequate biomarker result. An embedded process evaluation assessed factors influencing trial adoption, recruitment, and decision making. This study is registered with ISRCTN, ISRCTN65937227, and ClinicalTrials.gov, NCT01972425.FindingsBetween Nov 6, 2013, and Sept 13, 2016, 360 patients were screened for inclusion in the study. 146 patients were ineligible, leaving 214 who were recruited to the study. Four patients were excluded before randomisation, meaning that 210 patients were randomly assigned to biomarker-guided recommendation on antibiotics (n=104) or routine use of antibiotics (n=106). One patient in the biomarker-guided recommendation group was withdrawn by the clinical team before bronchoscopy and so was excluded from the intention-to-treat analysis. We found no significant difference in the primary outcome of the distribution of antibiotic-free days in the 7 days following bronchoalveolar lavage in the intention-to-treat analysis (p=0·58). Bronchoalveolar lavage was associated with a small and transient increase in oxygen requirements. Established prescribing practices, reluctance for bronchoalveolar lavage, and dependence on a chain of trial-related procedures emerged as factors that impaired trial processes.InterpretationAntibiotic use remains high in patients with suspected ventilator-associated pneumonia. Antibiotic stewardship was not improved by a rapid, highly sensitive rule-out test. Prescribing culture, rather than poor test performance, might explain this absence of effect.FundingUK Department of Health and the Wellcome Trust.

412. Host Response Biomarkers Predict Clinical Failure in Patients with Staphylococcus aureus Bacteremia (SAB) Treated with Flucloxacillin (FLU) or Vancomycin (VAN)

BackgroundImbalance among innate mediators such as IL-1β, IL-10, and TNF-α portends poor outcomes of persistence and death in patients with SAB. Previous studies did not consider the role of antibiotic treatment in this important host–pathogen relationship. In this study of SAB, we determined cytokine signatures that correlate with the composite endpoint of clinical failure (bacteremia duration >4 days or 30-day mortality) in Australian patients treated with FLU or VAN.MethodsSera from 86 patients with SAB (24.4% MRSA) were obtained from a clinical study of patients treated with FLU or VAN. All of the patient samples were collected at clinical presentation (day 0 or day 1 of infection) and were treated with FLU or VAN throughout. Patients were classified into either clinical success (CS = 68) or clinical failure (CF = 18), defined as death or prolonged bacteremia >4 days. Patient demographic and infection characteristics were collected. A 10-multiplex TH1/TH2 cytokine analysis was performed using electrochemoluminescence with the Meso-Scale Discovery platform analyzed by Mann–Whitney U.ResultsPatients’ median values were significantly elevated and above the normal range in CF for IL-1β (P = 0.029), IL-10 (P = 0.018), TNF-α (P = 0.042), and IL-6 (P = 0.006) (figure). Epidural abscess source was associated with CF, but no other host or pathogen characteristics correlated to outcome. Patients infected with isolates with VAN MIC = 2 mg/L (by Etest and broth dilution) had lower concentrations of IL-1β and IL-10 (P = 1.5 mg/L. In ROC analysis, IL-1β, IL-10, TNF, and IL-6 were higher sensitivity and specificity predictors of CF (AUC 0.65–0.71; P = 0.05).ConclusionA suboptimal host immune response to SAB at presentation predicts adverse clinical outcomes. IL-10, TNF-α, and IL-6 serum concentrations appear to reflect immunopathology in patients with SAB. These predictive markers may be considered in therapeutic clinical decision-making, such as escalation of alternative therapies in high-risk patients and/or de-escalation treatment in low-risk patients. These data offer steps toward further refining therapeutic precision for patients with SAB beyond the standard clinical or microbiological metrics that are employed in current practice. Disclosures All authors: No reported disclosures.

Ovarian interleukin profile and pregnancy outcome in women undergoing assisted reproduction: A prospective study

Context: Ovarian interleukins (ILs) mediate folliculogenesis, gametogenesis, fertilization, embryo development and implantation. Aims: Evaluation of the role of the quantitative levels of follicular fluid (FF) IL-1Beta (1β), IL-10 and IL-12B (p-40 subunit) in women underwent assisted reproduction. Setting and Design: Prospective observational study conducted between July 2013 and August 2015 at University Hospital’s in vitro fertilization (IVF) set up in North India. Materials and Methods: Women (n = 168) were worked up for IVF/intracytoplasmic sperm injection-embryo transfer (ICSI-ET) cycles. FF samples were pooled and collected from ovarian follicles of size ≥16 mm for each woman on the day of oocyte retrieval. Quantitative levels of IL-1β, IL-10 and IL-12B were estimated by enzyme-linked immunosorbent assay technique. Quantitative levels along with demography, cycle characteristics, endometrial thickness, number of retrieved oocytes, fertilization rate and embryos quality were compared between pregnant and nonpregnant groups of women. Student T-test, Mann–Whitney U-test, chi-squares test and logistic regression were applied as appropriate. Statistical significance level was calculated at P Results: Women (n = 168) were divided into pregnant (Group A; n = 75) and nonpregnant (Group B; n = 93) groups. The median levels of IL-1β, IL-10 and IL-12B levels were found significantly lower in women in Group A as compared to Group B [41.2 pg/mL (1.7–370) vs. 78.5 (5.6–313.58); P Conclusion: Lower concentrations of IL-1β, IL-10 and IL-12B in FF were found in a protagonist with positive pregnancy outcome and can be served as a reliable predictive marker of successful IVF/ICSI-ET outcome in women underwent assisted reproduction.

EMMPRIN Is Secreted by Human Uterine Epithelial Cells in Microvesicles and Stimulates Metalloproteinase Production by Human Uterine Fibroblast Cells

Endometrial remodeling is a physiological process involved in the gynecological disease, endometriosis. Tissue remodeling is directed by uterine fibroblast production of matrix metalloproteinases (MMPs). Several MMPs are regulated directly by the protein extracellular matrix metalloproteinase inducer (EMMPRIN) and also by proinflammatory cytokines such as interleukin (IL)1-α/β. We hypothesized that human uterine epithelial cells (HESs) secrete intact EMMPRIN to stimulate MMPs. Microvesicles from HES cell-conditioned medium (CM) expressed intact EMMPRIN protein. Treatment of HES cells with estradiol or phorbyl 12-myristate-13-acetate increased the release of EMMPRIN-containing microvesicles. The HES CM stimulated MMP-1, -2, and -3 messenger RNA levels in human uterine fibroblasts (HUFs) and EMMPRIN immunodepletion from HES-cell concentrated CM reduced MMP stimulation (P < .05). Treatment of HUF cells with low concentrations of IL-1β/α stimulated MMP production (P < .05). These results indicate that HES cells regulate MMP production by HUF cells by secretion of EMMPRIN, in response to ovarian hormones, proinflammatory cytokines as well as activation of protein kinase C.

Is there a role for neuronal nitric oxide synthase (nNOS) in cytokine toxicity to pancreatic beta cells?

Nitric oxide (NO), produced by the action of the inducible NO synthase, plays a crucial role in cytokine toxicity to pancreatic beta cells during type 1 diabetes development. It was the aim of this study to analyze the role of the neuronal NOS (nNOS) in proinflammatory cytokine-mediated beta cell toxicity. Expression of different isoforms of nitric oxide synthase in insulin-secreting INS1E cells and rat islets was analyzed by quantitative real-time PCR and Western blotting. The expression of nNOS in insulin-secreting INS1E cells was similar to that found in rat brain, while two other isoforms, namely the endothelial eNOS and inducible iNOS were not expressed in untreated cells. IL-1β alone or in combination with TNF-α and/or IFNγ induced iNOS but not eNOS expression. In contrast, nNOS expression was strongly decreased by the mixture of the three proinflammatory cytokines (IL-1β, TNF-α and IFNγ) both on the gene and protein level in INS1E cells and rat islet cells. The effects of cytokines on glucose-induced insulin-secretion followed the pattern of nNOS expression reduction and, on the other hand, of the iNOS induction. The data indicate that a low level of nitric oxide originating from the constitutive expression of nNOS in pancreatic beta cells is not deleterious. In particular since proinflammatory cytokines reduce this expression. This nNOS suppression can compensate for NO generation by low concentrations of IL-1β through iNOS induction. Thus, this basal nNOS expression level in pancreatic beta cells represents a protective element against cytokine toxicity.

Neuronal apoptosis by HIV-1 Vpr: contribution of proinflammatory molecular networks from infected target cells

BackgroundHuman immunodeficiency virus type 1 (HIV-1) induces neuronal dysfunction through host cellular factors and viral proteins including viral protein R (Vpr) released from infected macrophages/microglia. Vpr is important for infection of terminally differentiated cells such as macrophages. The objective of this study was to assess the effect of Vpr in the context of infectious virus particles on neuronal death through proinflammatory cytokines released from macrophages.MethodsMonocyte-derived macrophages (MDM) were infected with either HIV-1 wild type (HIV-1wt), Vpr deleted mutant (HIV-1∆Vpr) or mock. Cell lysates and culture supernatants from MDMs were analyzed for the expression and release of proinflammatory cytokines by quantitative reverse transcription-PCR and enzyme-linked immunosorbent assay respectively. Mitogen-activated protein kinases (MAPK) were analyzed in activated MDMs by western blots. Further, the effect of Vpr on neuronal apoptosis was examined using primary neurons exposed to culture supernatants from HIV-1wt, HIV-1∆Vpr or mock-infected MDMs by Annexin-V staining, MTT and Caspase - Glo® 3/7 assays. The role of interleukin (IL)-1β, IL-8 and tumor necrosis factor (TNF)-α on neuronal apoptosis was also evaluated in the presence or absence of neutralizing antibodies against these cytokines.ResultsHIV-1∆Vpr-infected MDMs exhibited reduced infection over time and specifically a significant downregulation of IL-1β, IL-8 and TNF-α at the transcriptional and/or protein levels compared to HIV-1wt-infected cultures. This downregulation was due to impaired activation of p38 and stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK) in HIV-1∆Vpr-infected MDMs. The association of SAPK/JNK and p38 to IL-1β and IL-8 production was confirmed by blocking MAPKs that prevented the elevation of IL-1β and IL-8 in HIV-1wt more than in HIV-1∆Vpr-infected cultures. Supernatants from HIV-1∆Vpr-infected MDMs containing lower concentrations of IL-1β, IL-8 and TNF-α as well as viral proteins showed a reduced neurotoxicity compared to HIV-1wt-infected MDM supernatants. Reduction of neuronal death in the presence of anti-IL-1β and anti-IL-8 antibodies only in HIV-1wt-infected culture implies that the effect of Vpr on neuronal death is in part mediated through released proinflammatory factors.ConclusionCollectively, these results demonstrate the ability of HIV-1∆Vpr to restrict neuronal apoptosis through dysregulation of multiple proinflammatory cytokines in the infected target cells either directly or indirectly by suppressing viral replication.

Inflammatory Cytokine IL-1β Upregulates Hepcidin Expression in Human Hepatoma Cell Lines.

The anemia of chronic disease (ACD) is commonly observed in patients with inflammatory disorders, malignancies, and chronic infections. ACD is characterized by low serum iron concentration and elevated serum ferritin concentration. A number of previous findings suggest that impaired mobilization of iron from the reticuloendotherial system (RES), such as macrophages, and an inhibition of iron absorption from intestine contribute to ACD. However, the precise mechanisms for generating the conditions of ACD have long been unclear despite extensive investigations until hepcidin was recently found. Hepcidin was found as one of antimicrobial peptides, but hepcidin is now thought to be a key regulator in iron metabolism. Hepcidin inhibits iron absorption from enterocytes and iron efflux from RES, which have suggested that hepcidin could account for the pathogenesis of ACD. Some cytokines have also been shown to modulate iron metabolism in the condition of inflammation. Interleukin-6 (IL-6), one of inflammatory cytokines, has been reported to induce hepcidin production not only in vitro but also in vivo. However, it is not clear whether other cytokines have such effect or not. We, therefore, investigated the possibility that other inflammatory cytokines might have a regulatory effect on hepcidin expression. Because hepcidin is produced mainly by hepatocytes, we used human hepatoma-derived cell line HuH-7 cells and hepatoblastoma-derived cell line HepG2 cells. Cells are incubated with or without inflammatory cytokines, such as IL-1β, IL-6, tumor necrosis factor α (TNFα), and then total RNA was extracted. Quantitative RT-PCR was then performed, revealing that IL-1β upregulate hepcidin mRNA expression as well as IL-6, although TNFα down regulates hepcidin expression in both cell lines. We next investigated the dose-response effect of IL-1β and IL-6 on hepcidin mRNA expression. Strong induction of hepcidin mRNA by IL-1β was observed when cells were incubated with low concentrations of IL-1β (0.2 ng/ml), although much higher concentrations of IL-6 were needed for hepcidin mRNA upregulation. It was likely that the concentrations of IL-1β that needed for hepcidin upregulation were more physiological than those of IL-6. Since it has been reported that IL-1β induce IL-6 production in hepatocytes, there was a possibility that the effect of IL-1β on hepcidin mRNA expression was not direct but come from IL-6 induced by IL-1β. However, the manners of IL-6 mRNA induction and hepcidin mRNA induction by IL-1β stimulation were quite different when cells were incubated with IL-1β at the concentrations ranging from 0.1 to 10 ng/ml. Moreover, antibody against IL-6 did not inhibit the induction of hepcidin mRNA by IL-1β stimulation. Therefore, it is likely that the effect of IL-1β on hepcidin mRNA expression is independent from that of IL-6. We conclude that inflammatory cytokine IL-1β can induce hepcidin expression and might be a key cytokine in the condition of ACD as well as IL-6, and IL-1β might be more important than IL-6 in physiological situations.

Keeping cool in acute liver failure: Rationale for the use of mild hypothermia

Encephalopathy, brain edema and intracranial hypertension are neurological complications responsible for substantial morbidity/mortality in patients with acute liver failure (ALF), where, aside from liver transplantation, there is currently a paucity of effective therapies. Mirroring its cerebro-protective effects in other clinical conditions, the induction of mild hypothermia may provide a potential therapeutic approach to the management of ALF. A solid mechanistic rationale for the use of mild hypothermia is provided by clinical and experimental studies showing its beneficial effects in relation to many of the key factors that determine the development of brain edema and intracranial hypertension in ALF, namely the delivery of ammonia to the brain, the disturbances of brain organic osmolytes and brain extracellular amino acids, cerebro-vascular haemodynamics, brain glucose metabolism, inflammation, subclinical seizure activity and alterations of gene expression. Initial uncontrolled clinical studies of mild hypothermia in patients with ALF suggest that it is an effective, feasible and safe approach. Randomized controlled clinical trials are now needed to adequately assess its efficacy, safety, clinical impact on global outcomes and to provide the guidelines for its use in ALF.

Measurement of cytokines and chemokines in parotid saliva from HIV+ patients

Objective. Salivary cytokines and chemokines may reflect the local immunity to microorganisms as well as any ongoing salivary gland dysfunction. In the past, cytokine profiles in human whole saliva have been proposed to assess oral mucosa immunity during HIV infection. Studies have shown that alterations in the expression of cytokines in glandular saliva are associated with candidal infection in HIV patients. Currently, an extensive cytokine and chemokine profile in saliva has not been established. Recent development of a multiplexed fluorescent microsphere-based immunoassay (xMAP technology) allows quantifying multiple proteins simultaneously using a very small amount of sample. The purpose of this study is to (1) establish a multiplexed assay using xMAP technology to measure cytokines and chemokines in saliva samples and (2) compare salivary cytokine and chemokine profiles between saliva samples from healthy subjects and HIV(+) patients. Study design. The Luminex 100 system is an xMAP technology and consists of a 10 × 10 matrix with a 100-analyte capacity. A set of cytokine and chemokine antibody–coupled fluorescent beads was used for the quantification of citrate-stimulated parotid saliva from HIV+ (n = 4; CD4+ counts < 200 cells/μL; salivary Candida count 500-9,000 cfu/mL) and healthy HIV− (n = 4; Candida 0 cfu/mL) subjects. The cytokines—interleukins (IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL10, IL-12, IL-13, and IL-15), interferon (IFN-γ), TNF-α, and GM-CSF—and the chemokines Eotaxin, RANTES, MCP-1, and MIP1α were assayed using 10 μL of saliva. Results. All 16 cytokines and 4 chemokines studied could be measured in detectable levels (>1 pg/mL). Overall, parotid saliva showed high concentrations of IFN-γ and Eotaxin (>200 pg/mL) and low concentrations of IL-1β, IL-13, and GM-CSF (<10 pg/mL). The concentrations of Eotaxin, GM-CSF, IL-1α, IL-1β, IL-2, IL-12p40, and IL-12p70 were elevated in stimulated parotid saliva from HIV+ patients compared to healthy HIV− subjects. The profiles for pg/mg total protein are similar to concentrations. Furthermore, 2 patients with abnormally low saliva secretions (<0.2 mL/min) had higher IL-2 and GM-CSF levels compared to patients with normal secretions (>0.2 mL/min). Conclusions. This is the first report that analytes can be measured in saliva using a multiplexed fluorescent microsphere-based platform, indicating that xMAP technology can assess salivary cytokine profiles. Although the number of subjects studied was small, the trends observed in this study suggest different salivary cytokine profiles for patients with HIV infection with a significant oral candidal burden and HIV− controls. The elevated levels of inflammatory cytokines and chemokines in saliva from HIV+ patients may reflect an ongoing immune response to a pathogen or microorganism. The increased levels of IL-2 in patients with low saliva production suggest the presence of T lymphocytes in the parotid gland.

Fetal plasma MMP-9 concentrations are elevated in preterm premature rupture of the membranes

Objective: The objective of this study was to determine whether the concentrations of matrix metalloproteinase-9 (MMP-9) in the fetal (fetal plasma and amniotic fluid) and maternal compartments (plasma) are different in patients presenting with preterm premature rupture of membranes (PROM) than in those with preterm labor and intact membranes. Study Design: Fetal plasma MMP-9, interleukin-1β (IL-1β), IL-6, soluble tumor necrosis factor receptors 1 (sTNF-R1) and 2 (sTNF-R2) were measured in fetuses with preterm labor and intact membranes (n = 96) and preterm PROM (n = 43). The concentrations of analytes were determined with sensitive and specific immunoassays. A P value <.05 was considered significant. Results: (1) The median fetal plasma MMP-9 concentration was significantly higher in fetuses with preterm PROM than in those with preterm labor (P =.035). (2) In contrast, fetal plasma IL-1β, sTNF-R1, and sTNF-R2 were significantly higher in patients with preterm labor than in those with preterm PROM (IL-1β, P =.01; sTNF-R1, P =.003; and sTNF-R2, P =.02). (3) The median amniotic fluid concentration of MMP-9 was higher in patients with preterm PROM than in those with preterm labor (P <.001). Conclusion: Fetuses with preterm PROM have increased concentrations of an enzyme (MMP-9) implicated in the mechanism of membrane rupture but lower concentrations of IL-1β, sTNF-R1, and sTNF-R2 than fetuses with preterm labor and intact membranes. A role for the fetus in the genesis of preterm PROM deserves consideration. (Am J Obstet Gynecol 2002;187:1125-30.)

Balance of synovial fluid IL-1β and IL-1 receptor antagonist and recovery from Lyme arthritis

Borrelia burgdorferi, the causative agent of Lyme disease, is a potent inducer of interleukin-1β (IL-1β), a cytokine implicated in the pathogenesis of inflammatory arthritis. The balance between IL-1 and the IL-1 receptor antagonist (IL-1ra), a naturally occurring inhibitor of IL-1, might influence disease expression. To explore this possibility, we have done a retrospective study that compared the clinical course of Lyme arthritis in 83 patients with concentrations of IL-1β and IL-1ra in the patients' synovial fluid. Patients with high concentrations of IL-1ra and low concentrations of IL-1β had rapid resolution of attacks of arthritis, whereas patients with the reverse pattern of cytokine concentrations had long intervals to recovery. Thus, the balance between synovial fluid IL-1β and IL-1ra concentrations relates to the time to recovery from an episode of Lyme arthritis.