BACKGROUND

The Infectious Systemic Inflammatory Response syndrome and multiple organic dysfunction have common physiopathological mechanisms. Multiple organic dysfunction can be assessed using severity scores.

OBJECTIVE

To relate cytokine kinetics with a multiple organic dysfunction score during sepsis.

METHODS

Tumor necrosis factor alpha (TNF alpha) and interleukin 6 (IL6) kinetics were studied in 25 patients with severe sepsis with less than 48 h of evolution and interleukin 1 beta (IL beta) kinetics was studied in 13 patients. Measurements were made at 0, 12, 24 and 48 hours after admission to the study, using an ELISA technique. These parameters were correlated with the Marshall multiple organic dysfunction score and survival.

RESULTS

Mean age of study subjects was 70 years, the APACHE II score was 16.9 +/- 6 and the Marshall score was 6.8 +/- 3.6. Sepsis was of pulmonary origin in 56% of patients and intra abdominal in 32%. Mortality was 36%. TNF alpha increased during the study period (24.1 pg/ml initially and 37.8 pg/ml at 24 hours, with a slight posterior reduction, p < 0.02). These levels had no association with mortality or organic dysfunction. IL6 remained elevated during the first hours and had a tendency to decrease thereafter. Decreased patients had higher values than survivors (306 pg/ml and 55.4 pg/ml respectively, p = 0.011). Its values were tightly correlated with Marshall score, with the number of failing organs, with the presence of shock and with probability of dying during hospitalization. IL1 beta remained low and was not associated with clinical parameters.

CONCLUSIONS

There is a tight correlation between the elevation of IL6 and the severity of the Systemic Inflammatory Response and mortality in these patients with sepsis.

We analyzed the proliferative response of DA1, DA1-a, NFS 60, 32DC1, Ea 3-17 and FDCP2 murine interleukin 3 (mIL3)-sensitive cell lines to human recombinant IL1 alpha, IL1 beta, IL2, IL4, IL5, IL6, granulocyte-CSF (G-CSF), macrophage-CSF (M-CSF), granulocyte-macrophage-CSF (GM-CSF), interferon gamma (IFN gamma), tumor necrosis factor (TNF) alpha, gibbon IL3, purified HILDA and mouse recombinant IL3, IL4, GM-CSF. All cell lines responded to mIL3 and IL4. Various response patterns were observed with human IL2, G-CSF and murine GM-CSF. Human IL4 and GM-CSF were absolutely inefficient in triggering proliferation of lines sensitive to their murine counterparts, confirming the species specificity of these two cytokines. Among the cell lines responding to G-CSF, some variations in sensitivities were observed. Thus the 32DC1 cell line needed 10 to 30 times more G-CSF to reach the same level of proliferation as the NFS60 cell line. DA1-a was the only factor-dependent cell line responding to purified HILDA, and IL6 was found to trigger proliferation of the NFS60 cell line, with a half-maximum effect observed for 0.1 pM of hormone.

It has been shown that the inflammatory cytokine tumor necrosis factor α (TNFα) plays a role in the development of hypertension and end-stage renal diseases. We hypothesize that TNFα contributes to endothelial dysfunction and cardiac and vascular injury in deoxycorticosterone acetate (DOCA)/salt-hypertensive mice. The wild-type or TNFα-deficient mice were uninephrectomized and implanted with DOCA pellet treatment for 5 weeks; the mice were given either tap water or 1% NaCl drinking water. DOCA mice developed hypertension (systolic blood pressure (SBP): 167 ± 5 vs. 110 ± 4 mmHg in control group, p < 0.05), cardiac and vascular hypertrophy, and the impairment of endothelium-dependent relaxation to acetylcholine (EDR). TNFα deficiency improved EDR and lowered cardiac and vascular hypertrophy with a mild reduction in SBP (152 ± 4 vs. 167 ± 5 mmHg in DOCA group, p < 0.05) in DOCA mice. The mRNA expressions of the inflammatory cytokines, including TNFα, interleukin 1β (IL1β), monocyte chemotactic protein 1 (MCP1), and monocyte/macrophage marker F4/80 were significantly increased in the aorta of DOCA-hypertensive mice; TNFα deficiency reduced these inflammatory gene expressions. DOCA-hypertensive mice also exhibited an increase in the vascular oxidative fluorescence intensities, the protein expressions of gp91phox and p22phox, and the fibrotic factors transforming growth factor β and fibronectin. TNFα deficiency reduced oxidative stress and fibrotic protein expressions. The DOCA mice also showed a decrease in the protein expression of eNOS associated with increased miR155 expression; TNFα deficiency prevented a decrease in eNOS expression and an increase in miR155 expression in DOCA mice. These results support the idea that TNFα significantly contributes to vascular inflammation, vascular dysfunction, and injury in hypertension.

Antibody response to an antigen involves the co-operation between three types of cells: macrophages, T cells and B cells. The cognate interactions between these cells play a fundamental role in the expression of a specific antibody response, but the last is modulated by antigen-nonspecific soluble factors produced either by macrophages or by T cells. Macrophages elaborate a spectrum of molecules modulating the function of lymphoid cells; among them are IL1 and prostaglandins of the E series, which are respectively enhancer and inhibitor of the antibody response in vitro. These molecules alter T cell and B cell activities through different mechanisms involving activation or inhibition of IL2 production, or alteration of cells surface antigens. However, the cellular events following the fixation of soluble factor on its receptors are not known.

The mammalian oviduct is an essential site for sperm storage, the transport of gametes, fertilization, and embryo development-functions that are aided by cytokines secreted from oviduct epithelial cells (OECs). Aging leads to cellular and organ dysfunction, with infertility associated with advanced maternal age. Few studies have investigated age-dependent changes in the oviduct as a possible cause of infertility, so we compared OECs from young (30-50 months) versus aged (more than 120 months) cattle. Next-generation sequencing was first used to identify age-related differences in gene expression. Several proinflammatory-related genes (including IL1B, IL1A, IL1BGN, and LUM) were down-regulation in aged OECs. Indeed, IL1 B and IL8 abundance was higher in aged OECs than in young OECs. Young OECs also tended to proliferate faster, and the revolution frequency of young, ciliated OECs was higher than that of their aged counterparts. In contrast, aged OECs possessed more F-actin, an actin cytoskeleton marker associated with reduced elasticity, and contained high levels of reactive oxygen species, which are mediators of inflammation and senescence. These different functional characteristics of bovine OECs during the post-ovulatory phase support the emerging concept of "inflammaging," that is, age-dependent inflammation. Mol. Reprod. Dev. 83: 815-826, 2016 © 2016 Wiley Periodicals, Inc.

The potentialities of a new non-invasive optical scanning microscopy technique were evaluated through 3D analysis of chondrocyte-matrix interactions. Five different 2D or 3D culture systems were used: (1) MonoLayer (ML) of human chondrosarcoma cell line; (2) rat or human chondrocytes encapsulated in Alginate Bead (AB); (3) human chondrocytes encapsulated in Alginate Sponge (AS); (4) Rat Femoral Head Cap (RFHC); (5) slices of knee human Osteoarthritic Cartilage (HOAC). Chondrocytes ML, AB, RFHC were incubated for 24 h in vitro in the presence of recombinant human interleukin1-beta (rhIL1-beta) and the effects on cytoskeleton organisation (F-actin filament), Focal Adhesion Kinase (FAK) expression (tyrosine kinase), collagenase B expression (metalloprotease) were studied. Furthermore, the production of intracellular IL1-beta by LPS- or rhIL1-beta-stimulated chondrocytes was shown to be partly suppressed by rhein (active metabolite of diacerhein) in all culture systems. This high resolution light microscopy gave complementary information that could be important for a better understanding of the interaction of chondrocytes with the extracellular matrix in a variety of culture devices.

Incubation of C6 astrocytoma cells with bacterial endotoxin (lipopolysaccharide; LPS) plus interferon-gamma (IFN-gamma), or with a combination of cytokines (TNF-alpha, IL1-beta, and IFN-gamma) leads to high levels of inducible nitric oxide synthase (iNOS) expression. Previous results demonstrated a requirement for tyrosine kinase (TK) activities for iNOS induction. In the present study, a set of structurally related TK inhibitors, the tyrphostins (TYRs), were used to characterize possible differences between LPS and cytokine iNOS induction. All TYRs tested suppressed both types of induction. However, dose-response curves revealed significant differences in the IC50 values obtained for some TYRs (T25 and T56), and significant differences in the IC50 potency rank order when comparing inhibition of LPS versus cytokine-dependent iNOS induction. These results are consistent with differential TK utilization by the LPS versus cytokine pathways of iNOS induction, and establish a basis for developing further selective inhibitors of iNOS expression.

Tumor necrosis factor (TNF) and interleukin 1 (IL1) activate a protein kinase, TIP kinase, which phosphorylates beta casein in vitro. We have now identified its main phosphorylation site on beta casein, Ser124 (Km approximately 28 mu M), and a minor phosphorylation site, Ser142 (Km approximately 0.7 mM). The sequence motif that determined the phosphorylation of Ser124 by the kinase was studied with synthetic peptides bearing deletions or substitutions of the neighboring residues. This allowed synthesis of improved substrates (Km approximately 6 mu M) and showed that efficient phosphorylation of Ser124 was favored by the presence of large hydrophobic residues at positions +1, +9, +11, and +13 (counted relative to the position of the phosphoacceptor amino acid) and of a cysteine at position -2. Peptides in which Ser124 was replaced by tyrosine were also phosphorylated by TIP kinase, showing it to have dual specificity. It is unable to phosphorylate the MAP kinases in vitro and is therefore not directly involved in their activation. Its biochemical characteristics indicate that TIP kinase is a novel dual specificity kinase, perhaps related to the mixed lineage kinases. It copurified with a phosphoprotein of about 95 kDa, which could correspond either to the autophosphorylated kinase or to an associated substrate.

Bovine nonfat dry milk (NDM) and major whey components (lactose, alpha-lactalbumin, and beta-lactoglobulin) were evaluated for their effects on IL-6 and IL-8 production in human intestinal-like Caco-2 cells unstimulated or stimulated with IL-1beta. All the whey components investigated and NDM induced IL-6 production by Caco-2 cells; the most significant increase was observed with beta-lactoglobulin. In the case of IL-1beta-stimulated cells, neither NDM nor the major whey components investigated contributed to the induction of IL-6 production after they were stimulated. Induction of IL-8 production by both alpha-lactalbumin and beta-lactoglobulin was higher than that by lactose and NDM; alpha-lactalbumin was a more potent inducer of IL-8 than beta-lactoglobulin and IL-1beta alone in both unstimulated and stimulated cells. In Caco-2 cells that were stimulated with IL1-beta, NDM and all the major whey components investigated had a synergistic effect on induction of IL-8 production, indicating that IL-8 induction was amplified by prior stimulation of cells by IL-1beta. This synergistic effect was not observed with IL-6. Our results suggest that immunomodulatory properties of milk components may be affected by other complex events in the gut.

Among the various candidate genes predisposing for cardiovascular diseases, HLA-DRB1* and IL-1β +3953C/T alleles have been implicated repeatedly. To test these in South India, we carried out a case control study of 323 Coronary Artery Disease (CAD) patients, 56 Rheumatic Heart Disease (RHD) patients and 254 endemic controls. The polymorphisms were studied by PCR - SSP and ARMS-PCR methods and results analyzed for various clinical and demographic parameters. In CAD, HLA-DRB1*14 allele showed significant predisposition (OR: 2.19; 95% CI: 1.04-4.58; p value=0.023), particularly in male patients (OR: 4.07; 95% CI: 1.20-13.81; p value=0.01) and further in males with Triple Vessel Disease (OR: 5.49; 95% CI: 1.45-20.60; p value=0.007). On the other hand, HLA-DRB1*15 predisposed for RHD (OR: 3.56; 95% CI: 1.87-6.78; p value=0.001) in both the genders. Population stratification showed this higher risk association in Vanniyar caste (OR: 5.00; 95% CI: 1.27-19.59; p value=0.022). Among the IL1-β +3953C/T polymorphism, the ancestral allele 'C' showed a significant high risk association with CAD (OR: 1.83; 95% CI: 1.24-2.70; p value=0.001), particularly in Mudaliar (OR: 6.07; 95% CI: 1.77-20.74; p value=0.003; AF=0.7) and Vanniyar castes (OR: 3.67; 95% CI: 0.92-14.57; p value=0.05; AF=0.660). Two different cardiac ailments studied, RHD & CAD thus showed varied associations in this South Indian cohorts. RHD having an infectious aetiology shared a HLA-DRB1*15 high risk association, while HLA-DRB1*14 and IL-1β +3953C predisposed for CAD, an inflammatory disorder, reiterating the diverse genetic predisposition of the two cardiac ailments studied.

OBJECTIVE

To evaluate CD4(+) CD28(+) and CD8(+) T-cell genes and the gene expression of IFN-γ, TNF-α, IL-1-β, IL-17A, IL-10, CCL-2/MCP-1, CCL-4, CCL-5 (RANTES), CXCR4, CCR5 and RANKL from cells in the periapical interstitial fluid from root canal infections in healthy patients (HIV-) and HIV-positive individuals (HIV+).

METHODS

Subjects included 20 HIV- and 23 HIV+ patients referred to the School of Dentistry at the Universidade Federal de Minas Gerais (Belo Horizonte, MG, Brazil). Almost all HIV+ patients were undergoing highly active antiretroviral therapy (HAART). Clinical samples were taken from teeth with pulp necrosis, and no patients had acute periapical symptoms at the time of the appointments. After cleaning and drying, 3 paper points were introduced into the root canal, passing passively through the root apex (2 mm) into the periapical tissues for 1 min. The samples were collected immediately after root canal cleaning and 7 days later (restrained root canal bacterial load) to characterize those gene expressions using real-time PCR.

RESULTS

Significantly higher levels of CD4(+) CD28(+) and CD8(+) T cells in teeth with restrained bacterial loads (second collection) compared with the first collection were observed in both HIV- and HIV+ samples. In HIV- patients, an increase in IL-10 and CXCR4 expression was demonstrated as well as a decrease in pro-inflammatory cytokines such as RANKL, IFN-γ, IL1-β and CCL5. However, in HIV+ patients an increase in cytokines IFN-γ, IL-1-β, TNF-α and IL-17A, and chemokines CCL-2, CXCR4 and CCR5 were observed. The chemokine CCL-5 was not detected in HIV+ individuals.

CONCLUSIONS

These findings suggest that after reducing the root canal bacterial load in HIV- individuals an anti-inflammatory response is generated whilst in HIV+ patients a pro-inflammatory response is sustained in the periapical area.

We have investigated the sequence of signals provided by a B- and null-cell-derived prothymocyte-differentiating activity (PTDA), phytohemagglutinin (PHA), and interleukin 2 (IL2) to the generation of mature T-lymphocytes by T-depleted bone marrow (BM) cells. Sequential studies show that preincubation of CD2-, CD3-, CD4-, CD6-, and CD8- BM cells with PTDA, but not with recombinant (r) IL2 or PHA increased their capacity to proliferate in liquid culture and to form agar T-cell colonies provided both PHA and rIL2 were added to the cultures. In contrast, the growth of T-cell-containing BM was significantly enhanced in both liquid and agar culture following its preincubation with rIL2 as well as with PTDA. The selective effect of PTDA on CD2-, CD3-, CD4-, CD6-, CD8- BM cells was abolished by adding a CD7 monoclonal antibody to the T-cell-purging coctail. Cell marker studies performed on T-cell-depleted BM-derived liquid or agar cultures have shown that they contain up to 70%-85% CD2+, CD3+, CD4+, CD8- cells. No IL1 or IL2 could substitute for PTDA, nor have these activities, as well as interferon (IFN), IL3, IL4, or granulocyte-macrophage colony-stimulating activity (GM-CSA) been detected so far in PTDA-containing preparations. These results indicate that PTDA can trigger marrow T-cell precursors into PHA-responsive T cells, which, following activation by PHA, require IL2 for growth. It is suggested that this may represent a thymus-independent alternative pathway for T-cell differentiation and activation.

OBJECTIVE

The aim of this study was to determine the localization of the rate-limiting enzyme indoleamine 2, 3-dioxygenase (IDO) and its metabolite in mice kidneys after adriamycin (ADR)-induced renal failure. We also examined the effect of L-tryptophan (Trp) administration on this model.

METHODS

BALB/c mice were treated with 15 mg/kg ADR to induce renal failure. The change of IDO and L-kynurenine (Kyn) following ADR-induced renal failure was examined by immunostaining combined with hematoxylin and eosin (HE) and Periodic acid-Schiff (PAS) staining for morphological analysis, and the concentration of L-Kyn was measured by HPLC. The effect of L-Trp administration on ADR-induced renal failure was also investigated.

RESULTS

Time-dependent increase of IDO immunostaining was observed in tubular cells from Day 4 after ADR injection. It was found that mice fed with L-Trp had less chance of renal failure at four days after ADR injection than mice untreated with L-Trp, but not at seven days. Further, L-Trp treatment significantly suppressed an increased level of TNF-alpha mRNA, but not of TGF-beta and IL1-beta after renal injury.

CONCLUSIONS

Local IDO expression in tubules was induced markedly after ADR- induced renal failure. L-Trp administration can be effective in suppressing ADR-induced renal failure at an early stage.

Epstein-Barr virus (EBV), in addition to its transforming properties, contributes to the pathogenesis of several inflammatory diseases. Here, we investigated its involvement in oral lichen planus (OLP), a common autoimmune-like disease of unknown etiopathogenesis that can display a malignant potential. EBV-infected cells (EBV+ cells) were sought in a large series of clinically representative OLPs ( n = 99) through in situ hybridization to detect small noncoding EBV-encoded RNAs. Overall, our results demonstrated that EBV was commonly found in OLP (74%), with significantly higher frequency (83%) in the erosive form than in the reticular/keratinized type mild form (58%). Strikingly, many erosive OLPs were massively infiltrated by large numbers of EBV+ cells, which could represent a large part of the inflammatory infiltrate. Moreover, the number of EBV+ cells in each OLP section significantly correlated with local inflammatory parameters (OLP activity, infiltrate depth, infiltrate density), suggesting a direct relationship between EBV infection and inflammatory status. Finally, we characterized the nature of the infiltrated EBV+ cells by performing detailed immunohistochemistry profiles ( n = 21). Surprisingly, nearly all EBV+ cells detected in OLP lesions were CD138+ plasma cells (PCs) and more rarely CD20+ B cells. The presence of EBV+ PCs in erosive OLP was associated with profound changes in cytokine expression profile; notably, the expression of key inflammatory factors, such as IL1-β and IL8, were specifically increased in OLP heavily infiltrated with EBV+ PCs. Moreover, electron microscopy-based experiments showed that EBV+ PCs actively produced EBV viral particles, suggesting possible amplification of EBV infection within the lesion. Our study thus brings conclusive evidence showing that OLP is commonly infiltrated with EBV+ PCs, adding a further puzzling element to OLP pathogenesis, given that PCs are now considered to be major regulatory immune cells involved in several autoimmune diseases (ClinicalTrials.gov NCT02276573).

OBJECTIVE

The purpose of the present investigation was to develop and validate two separate enzyme-linked immunosorbent assays (ELISA) for quantitation of exogenous human epidermal growth factor (hEGF1-53) and its truncated fragment (hEGF1-48) in rat plasma.

METHODS

The present assay systems were based on the sandwiching of the antigen between a monoclonal mouse anti-hEGF1-53 antibody, pre-coated on a 96-well polystyrene plate, and a polyclonal rabbit anti-hEGF1-48 antibody, which is then detected with a peroxidase-labeled goat anti-rabbit antibody.

RESULTS

The calibration curves for hEGF1-48 and hEGF1-53 in plasma were validated over a concentration range of 7.8-250 and 62.5-1000 pg/ml, respectively. Determined from replicate assays of hEGF1-48 quality control samples, the intra-assay precision and accuracy were < or = 8.8% RSD and within +/- 9.8%; and the inter-assay precision and accuracy were < or = 14.8% RSD and within +/- 9.7% RE, respectively. Determined from replicate assays of hEGF1-53 quality control samples, the intra-assay precision and accuracy were < or = 10.0% RSD and within +/- 8.5%; and the inter-assay precision and accuracy were < or = 10.0% RSD and within +/- 5.7% RE, respectively. The limit of quantitation of the hEGF1-48 and hEGF1-53 assay using 200 microliters plasma per well is 7.8 and 62.5 pg/ml, respectively. These two ELISA methods are specific to hEGFs and do not cross-react with mouse EGF or other growth factors (TGF alpha, TGF beta, PDGF, and FGF) or lymphokines (IL1 beta and TNF alpha). These validated methods have been routinely applied to assay of plasma samples from various pharmacokinetic studies in rats receiving intravenous hEGFs. Both assay methods were also adapted to assay endogenous hEGFs in biological fluids of different animal species.

CONCLUSIONS

Two sensitive ELISA methods have been validated for quantitation of hEGF1-53 and hEGF1-48 in rat plasma. Their utility has been demonstrated in the application of assaying immunoreactive concentration of exogenous and endogenous epidermal growth factors.

BACKGROUND

The benefits of antiretroviral therapy for HIV-infected subjects have been limited by an increased risk of metabolic and cardiovascular diseases. The objective of this study was to assess the effects of a low dose of marine omega-3 fatty acids on inflammatory marker concentrations in HIV-infected subjects under antiretroviral therapy (ART).

METHODS

This was a randomized, parallel, placebo-controlled trial that investigated the effects of 3 g fish oil/day (540 mg of eicosapentaenoic acid-EPA plus 360 mg of docosahexaenoic acid-DHA) or 3 g soy oil/day (placebo) for 24 weeks in 83 male and non-pregnant female HIV-infected adults on ART.

RESULTS

There were no differences between groups for the measures at baseline. Multilevel analyses revealed no statistically significant relationship between the longitudinal changes in high sensitivity-C reactive protein (hs-CRP) (Wald Chi2 = 0.17, p = 0.918), fibrinogen (Wald Chi2 = 3.82, p = 0.148), and factor VIII (Wald Chi2 = 5.25, p = 0.073) with fish oil. No significant changes in interleukin-6 (IL6), interleukin-1 beta (IL1-beta) and tumor necrosis factor-alpha (TNF-alpha) serum concentrations were observed with fish oil supplements for 12 weeks.

CONCLUSIONS

Compared to placebo, a low dose of 900 mg omega-3 fatty acids (EPA plus DHA) in fish oil capsules did not change hs-CRP, fibrinogen, factor VIII, IL6, IL1-beta and TNF-alpha serum concentrations in HIV-infected subjects on ART. Further investigations should consider the assessment of more sensitive inflammatory markers or higher doses to evaluate the effects of marine omega-3 fatty acids in this population. Registered at the Nederlands Trial Register, Identifier no. NTR1798.

OBJECTIVE

The levels of interleukin-1β (IL-1β), nitric oxide (NO), total antioxidant capacity (TAC), and total oxidant status (TOS) in gingival crevicular fluid (GCF) were determined during rapid maxillary expansion (RME) treatment.

METHODS

Fourteen patients (10-13 years old) were included. A modified hyrax appliance was used for the treatment. After periodontal parameters were recorded, GCF was collected from the first molars at each observation [T1:baseline:14 days after periodontal prophylaxis and instructions; T2:1 day later hyrax inserted, at passive position; T3:1 week later; after the first activation; T4:after 2 × 1/4 activation; T5:after 7 × 1/4 activation; T6:after 14 × 1/4 activation; T7:retention period on the 1 st month; and T8:retention period on the 3rd month].

RESULTS

Although the levels of IL1-β, NO, and PD increased significantly from T1 to T2, the GI, BOP%, and PI remained unchanged throughout treatment. GCF volume at buccal and palatal surfaces increased significantly from T1 to T4, T6, T7, and T8. The parameters in GCF and TAC levels were not only higher at palatal side in comparison with buccal, but also TOS levels increased at both buccal and palatal sides.

CONCLUSIONS

In this study, the differences of oxidative status and IL-1β levels during RME treatment could be attributable to orthopedic effect of the heavy forces on maxilla and minimal orthodontic forces on teeth applied by the RME apparatus.

Aim: Meat and its derivatives provide nutrients essential for human health. However, meat consumption, along with excessive fat intake, has been associated with gut inflammation, intestinal barrier dysfunction and alterations in gut microbiota. Herein, we investigated whether and how these changes in the intestinal barrier system affect the gut liver axis and hepatic injury and eventually lead to the progression of liver syndrome such as NAFLD.

Methods: Mice were fed with high fat (60% kcal) or low fat (12% kcal) along with soybean (control), chicken and pork proteins (HFCH, HFP, LFCH, and LFP) for 12 weeks. The biomarkers for liver injury were investigated after meat protein intake along with the high fat.

Findings: Greater amount of fat vacuoles visible in the H&E staining increased the inflammatory cell infiltration and disorganized liver structures were observed in the HFP-fed mice. Oil Red O staining revealed that the HFP-fed and HFCH-fed mice showed more lipid droplets, confirming the increased hepatic lipid accumulation. Potential serum markers for NAFLD, ALT and AST were increased in the HF meat diet groups. Key genes responsible for hepatic inflammation and lipogenesis, such as MCP-1, IL1-β and TNF-α were upregulated. HF meat protein diet-fed mice exhibited signs of compromised liver with increased levels of endotoxin in the liver and its binding protein in serum, upregulation of TLRs in the liver, and significant increase in TG, TC, LDL-C and HDL-C concentrations.

Significance: Intestinal inflammation and barrier dysfunction aggravate liver injury and fibrosis due to the intake of HF meat protein diets in mice, which may contribute to the progress of liver injury and associated complications. Gut inflammation may directly contribute to the development of NAFLD, especially of the gut vascular barricade dysfunction.

OBJECTIVE

To investigate the protective effect of Ambroxol on ventilator induced lung injury (VILI) of rats.

METHODS

30 healthy SD rats were randomly divided into three groups: group A was served as control group (n = 10, V(T) 8 ml/kg), group B received large V(T) ventilation (n = 10, V(T) 40 ml/kg), group C received large V(T) ventilation with pre-treatment of IP ambroxol. The contents of protein, TNF-alpha, IL1-beta, IL-8 in BALF and MDA, SOD, GSH levels in the homogenate of the rat lungs were assayed respectively. Total white blood cells in BALF were counted; The severities of lung injury of the three groups were assessed under a light microscope.

RESULTS

The albumin and total protein contents in BALF were significantly higher group A than in groups B and C. A large quantity of WBCs infiltration were found in BALF of group B, while the amount of WBCs were decreased in group C. TNF-alpha, IL-1beta and IL-8 level in BALF of group B were 101.6 pmol.L(-1).mg(-1) Pr +/- 37.4 pmol.L(-1).mg(-1) Pr, 0.47 microgram.L(-1).mg(-1) Pr +/- 0.14 microgram.L(-1).mg(-1) Pr and 2.02 microgram.L(-1).mg(-1) Pr +/- 0.49 microgram.L(-1).mg(-1) Pr, respectively. MDA, one of the parameters reflecting the extent of oxidative reaction, were produced profoundly in the lungs of group B, but the local levels of SOD, which reflects the ability of antioxidation, were decreased significantly in group B. There existed significant differences of the above parameters between group B and group A. Ambroxol could significantly decrease the locally produced TNF-alpha and inhibit the oxidative reactions, and increase the locally antioxidative potency. Severe acute lung injuries were detected in group A by histological examination and the extent of injury was much lighter in group C than in group B.

CONCLUSIONS

Locally increased inflammatory cytokines formation and the imbalanced oxidative/antioxidative reaction play important roles in the pathogenesis of VILI. Ambroxol has an obvious protective effect on VILI through its antioxidant/anti-inflammation potent.

Culture supernatants of tenosynovial tissues from patients with carpal tunnel syndrome undergoing chronic haemodialysis contained interleukin (IL) 1-like and IL6-like activity. These culture supernatants also induced active proliferation of rheumatoid synovial cells. Immunohistochemical analysis of teno-synovial tissues showed the accumulation of mononuclear cells bearing CD14 and HLA-DR antigens adjacent to the deposition of amyloid protein (beta 2 microglobulin). These cells also reacted with antibodies to IL1 and IL6 respectively. These data suggest that multiple cytokines, including IL1 and IL6, produced from tenosynovial tissues in patients with dialysis associated amyloidosis might induce the proliferation of synovial cells that, together with deposition of amyloid protein, might cause carpal tunnel syndrome.

We measured tumor necrosis factor (TNF alpha), interleukin-1 (IL1-B), and beta-2 microglobulin (BBBIL1-B (greater than 20 pg/ml) at least once, and 2 had detectable levels prior to all dialyses. Serum TNF alpha, IL1-B and BIL1-B or BIL1-B and TNF alpha are not uniformly observed in hemodialysis patients, arguing against a role for these substances as systemic uremic toxins.