Mohamed El-Boshy

Immunomodulatory effect of dietary Saccharomyces cerevisiae, β-glucan and laminaran in mercuric chloride treated Nile tilapia (Oreochromis niloticus) and experimentally infected with Aeromonas hydrophila.

Six hundred and forty Nile tilapia (Oreochromis niloticus) weighing 80-100g were randomly allocated into eight equal groups (80 each). The first group acts as control. Groups S, B and L were fed on a ration supplemented with Saccharomyces cerevisiae, beta-glucans and laminaran, respectively for 21 days. Groups M, MS, MB and ML were subjected throughout the experiment to sublethal concentration of mercuric chloride (0.05 ppm). Gps. MS, MB and ML were fed on a ration containing S. cerevisiae, beta-glucan and laminaran respectively for 21 days. Fish were challenged with Aeromonas hydrophila (0.4 x 10(7) cells mL(-1)) via intra-peritoneal injection and the mortality rate was recorded up to 10 day post-challenge. The non-specific defense mechanisms, cellular and humoral immunity, beside the total and differential leukocytic count were determined. Lymphocyte transformation index, phagocytic activity percent, phagocytic index, total lymphocyte count, serum bactericidal activity and nitric oxide as well as the survival rate were insignificantly changed after 21 day in gps. MS&ML, when compared with mercuric chloride immune depressed group M. These parameters as well as the neutrophil adhesion, serum nitric oxide and survival rate were significantly increased in gp. MB when compared with gp. M. Meanwhile the cellular and humoral immunity beside the survival rate were significantly increased in groups S, B, L when compared with control group. It could be concluded that the whole yeast S. cerevisiae, beta-glucan and laminaran can be used as immunostimulants for the farmed Nile tilapia. The beta-glucans could be used in farmed Nile tilapia, under immune depressive stressful condition to increase their resistance to diseases.

Protective effects of selenium against cadmium induced hematological disturbances, immunosuppressive, oxidative stress and hepatorenal damage in rats

Cadmium is a non-essential toxic metal used in industrial process, causes severe risk to human health. Selenium (Se) is an essential trace mineral of fundamental importance for human health. Selenium has antioxidant enzymes roles and is needed for the proper function of the immune system. In this study, the protective effects of selenium against cadmium intoxication in rats have been investigated by monitoring some selective cytokines (IL-1β, TNF α, IL-6, IL-10 and IFN-γ), antioxidant enzymes reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and lipid peroxidation malondialdehyde (MDA) as well as some selective biochemical markers of liver and kidney functions. Thirty-two rats were divided into four equal groups; the first group was used as a control. Groups 2-4 were treated with selenium (Se; 0.1mg/kg BW), cadmium (Cd; 40mg/L drinking water) and selenium plus cadmium, respectively. Rats were orally administered their relevant doses daily for 30 days. Blood samples were collected from heart puncture at the end of the experiment (30 days) for complete blood picture (CBC) and serum was separated to evaluate the different immunological parameters and biochemical parameters, as well as liver specimens for Cd and Se estimation. Rats in the Cd treated group have a significantly higher hepatic concentration of Cd than in other treated groups. Results revealed that cadmium significantly increased IL-1β, TNF α, IL-6 and IL-10, beside peripheral neutrophils count, while the IFN-γ and lymphocytes were decreased in rat sera. In addition, GSH level, CAT, SOD and GPx activities were significantly decreased while lipid peroxidation (MDA) was increased. Regarding, liver and renal markers, they were significantly increased in the activities of aminotransferases (AST, ALT), urea and creatinine, while total plasma proteins and albumin were significantly decreased. On the other hand, selenium treated group, showed significantly increased IFN-γ, GSH level, CAT, and GPx activities, as well as lymphocyte count while IL-10 was decreased. Selenium in combination with cadmium, significantly improved the elevation of serum IL-1β, IL-6, TNF α, IL-10 and malondialdehyde in addition to enhancing the antioxidant enzyme activities of GSH, CAT, GPx and SOD. Moreover, selenium has ameliorated the cadmium-induced liver and kidney damage by improving hepatic and renal markers. The results of this investigation demonstrated that selenium has the potential to countermeasure the immunosuppressive as well as hepatic and renal oxidative damage induced by cadmium in rats; selenium has shown promising effects against Cd toxicity.

The role of serum IL-17 and IL-6 as biomarkers of disease activity and predictors of remission in patients with lupus nephritis

AIM To determine the role of IL-17 and IL-6 in the pathogenesis of SLE as biomarkers of disease activity and predictors of remission and outcome of therapy in patients with active lupus nephritis. METHODS The study was carried out on 72 SLE female patients and 70 sex- and age-matched normal healthy subjects as controls. SLE disease activity was assessed in all patients with (SLEDAI-2k scores). Plasma levels of IL-6, and IL-17 were measured by enzyme linked immunosorbent assay and their levels were correlated with clinical manifestations of the disease and (SLEDAI-2k). ROC curve analysis was performed to determine the validity of both cytokines in prediction of activity and remission of active lupus nephritis. RESULTS SLE patients were found to have significantly higher levels of IL-17 (p<0.001) and IL-6 (p<0.001), in relation to normal subjects. Active group of patients had higher levels of both cytokines than the inactive one (P<0.001). Elevated serum levels of both cytokines were associated with active lupus nephritis, anemia and positively correlated with SLEDAI-2k scores (P=0.025 for IL-17 and P<0.001 for IL-6). There was a significant positive correlation between IL-6 and IL-17 serum concentrations during periods of disease activity (r=0.497, P=0.005) as well as during remission (r=0.662, P<0.001). ROC curve analysis for IL-6 and IL-17, as predictor of disease activity reviled, optimal cutoff level of 12.3 pg/ml and 19.7 pg/ml, with AUC=0.93, and 0.95, for both cytokines respectively, while as predictors of remission of active lupus nephritis, provide a cutoff value of IL-6 at 20.8 pg/ml, with AUC 0.80, and a cutoff value of IL-17 at 27.0 pg/ml, with AUC 0.82. CONCLUSION In conjunction with their major role in pathogenesis of SLE, baseline serum levels of IL-6 and IL-17 can be used as sensitive biomarkers for disease activity, as well as predictors of remission of lupus nephritis.

Traumatic Reticuloperitonitis in Water Buffalo (Bubalus bubalis): Clinical Findings and the Associated Inflammatory Response

The present study was carried out to describe the clinical picture of traumatic reticuloperitonitis (TRP) in water buffalo (Bubalus bubalis) and to evaluate the inflammatory and immunologic responses for this clinical condition. Twenty-two buffalo with acute local TRP were monitored in our study. Additionally, 10 clinically healthy buffalo were randomly selected and served as controls. Acute local TRP was initially diagnosed by clinical examination and confirmed by ultrasonographic (USG) examination and/or necropsy findings. Blood samples were collected from all examined buffalo to measure the respective levels of tumor necrosis factor alpha (TNF-α), interleukin (IL)-1β, IL-6, IL-10 and interferon gamma (INF)-γ, serum amyloid A (SAA), C-reactive protein (CRP), haptoglobin (Hp), fibrinogen (Fb), and serum sialic acid (SSA). It was found that TNF-α, IL-1β, IL-6, IL-10, SAA, CRP, Hp, Fb, and SSA were significantly higher in buffalo with TRP than the controls. Our findings suggest that the examined immunologic variables were helpful in documenting the inflammatory response in buffalo with TRP. However, their diagnostic usefulness only becomes apparent when considered in tandem with the clinical findings for any given animal, its anamnesis, and a subsequent USG assessment. Due to the frequent complications of TRP, more accurate indicators of its occurrence and severity would be useful.

The role of acute phase cytokines in the recovery and disease progress of Theileria annulata-infected cattle

The present study was conducted to evaluate the cytokine response following natural infection of Theileria annulata in cattle. Initial survey included 173 crossbred cattle which were examined for the presence of Theileria piroplasms. The investigated cattle were clinically and parasitologically examined. Blood samples were collected from all examined cattle for microscopic examination, PCR assays (using primers of Theileria spp., Babesia spp., and T. annulata), and cytokines measurement. It was found that 38 cattle were positive for the presence of at least one species of Theileria; meanwhile Babesia piroplasms were not detected either by microscopy or PCR assay. When T. Annulata-specific primers were used, 33 gave positive results. Twenty-two out of 33 T. annulata-infected cattle were only included in this study together with contemporaneous controls (n = 10). According to the severity of clinical signs, T. annulata-infected cattle were categorized into two groups; group 1 included ten cattle with mild clinical signs and group 2 included 12 cattle with overt clinical signs. Biochemically, tumor necrosis factor alpha, interleukin (IL)-1β, IL-6, IL-12, haptoglobin, and Fb were significantly higher (p < 0.05) in diseased cattle compared to control and in cattle of group 2 compared to those in group 1, while interferon gamma showed no significant variations between the two groups. We conclude that measurements of the pro-inflammatory cytokines following T. annulata infection provide a valuable and quantitative assessment of the response to infection. It seems likely that the pro-inflammatory cytokines play an important role in the host response to T. annulata. Our findings suggest that the pro-inflammatory cytokines are suitable markers of inflammatory reactions in T. annulata-infected cattle.

Prognostic significance of platelet count in SLE patients

Abstract Hematological abnormalities, especially thrombocytopenia (TCP), are highly prevalent among patients with systemic lupus erythematosus (SLE) and at the same time it has been reported as a significant prognostic factor of SLE course. We further investigate the correlation between platelet count and the clinical manifestations and disease activity of SLE, in a cohort of Saudi Arabian female patients. A retrospective analysis was done for the medical records of 100 SLE female patients, selected from all patients diagnosed and treated for SLE at the Rheumatology outpatient clinics in Hera’a General Hospital, Holly Makkah, Saudi Arabia. The data collected from every patient’s file included laboratory investigations (complete blood count, platelet parameters, ESR, anti-double-stranded DNA antibody, ANA), clinical manifestations, as well as SLE disease activity index (SLEDAI-2k) scores throughout a period of six sequential follow-up visits. Patients were divided into three groups according to the SLEDAI-2k: mild, moderate, and high-activity group. We found that, out of 100 patients, TCP was the most prevalent hematological abnormality evident in 15%, more than leucopenia (14%) and anemia (2%). TCP was acute in onset and associated with arthritis, neurologic manifestations, and nephritis. Platelet count showed a significant negative correlation with disease activity, in all of the three groups of patients. We concluded that platelet count has a negative correlation with disease activity in SLE patients, whatever the associated manifestations, and it should be considered as a prognostic factor, identifying patients with aggressive disease course.

Dietary fucoidan enhance the non-specific immune response and disease resistance in African catfish, Clarias gariepinus, immunosuppressed by cadmium chloride

Fucoidan is sulfated polysaccharide extracted from seaweed brown algae. This study was designed to evaluate the immunomodulatory effects and disease resistance of dietary fucoidan on catfish, Clarias gariepinus, immunosuppressed by cadmium. Three hundred and sixty African catfish, C. gariepinus, was allocated into six equal groups. The first group served as a control. Groups (F1 and F2) were fed on fucoidan supplemented ration at concentrations of 4 and 6g/kg diet respectively for 21 days. Groups (Cd, CdF1 and CdF2) were subjected throughout the experiment to a sub-lethal concentration of 5ppm cadmium chloride solution and groups (CdF1 and CdF2) were fed on a ration supplemented with fucoidan. Macrophages oxidative burst, phagocytic activity percentages and lymphocytes transformation index were a significant increase in the fucoidan-treated groups (F1 and F2), while serum lysozyme, nitric oxide and bactericidal activity were enhanced only in group (F2) when compared with controls. These parameters as well as absolute lymphocyte count and survival rate were significantly increased in group (CdF2) when compared with cadmium chloride immunosuppressed group (Cd). It could be concluded that the fucoidan can be used as immunostimulant for the farmed African catfish, C. gariepinus as it can improve its resistance to immunosuppressive stressful conditions.

Vitamin D alleviates lead induced renal and testicular injuries by immunomodulatory and antioxidant mechanisms in rats

This study measured the effects of vitamin D (VD) supplementation on the underlying molecular pathways involved in renal and testicular damage induced by lead (Pb) toxicity. Thirty two adult male Wistar rats were divided equally into four groups that were treated individually or simultaneously, except the negative control, for four weeks with lead acetate in drinking water (1,000 mg/L) and/or intramuscular VD (1,000 IU/kg; 3 days/week). Pb toxicity markedly reduced serum VD and Ca2+, induced substantial renal and testicular injuries with concomitant significant alterations in the expression of VD metabolising enzymes, its receptor and binding protein, and the calcium sensing receptor. Pb also significantly promoted lipid peroxidation and pro-inflammatory cytokines (IL-4 and TNF-α) in the organs of interest concomitantly with declines in several anti-oxidative markers (glutathione, glutathione peroxidase and catalase) and the anti-inflammatory cytokine, IL-10. The co-administration of VD with Pb markedly mitigated renal and testicular injuries compared with positive controls. This was associated with restoration of the expression of VD related molecules, promotion of anti-oxidative and anti-inflammatory markers, but tissue Pb concentrations were unaffected. In conclusion, this report is the first to reveal potential protective effects for VD against Pb-induced renal and testicular injuries via anti-inflammatory and anti-oxidative mechanisms.

Interferon alpha gene expression and serum level association with low vitamin D levels in Egyptian female patients with systemic lupus erythematosus

Background Patients with systemic lupus erythematosus (SLE) are prone to develop vitamin D (25(OH) D3) deficiency, due to several factors and there is an association between lower vitamin D levels and higher SLE disease activity. The aim of this research was to assess the prevalence of vitamin D deficiency in Egyptian female patients with SLE. Furthermore, we analyzed the potential relationship between this deficiency and SLE manifestations, disease activity, and its effect on interferon alpha (IFN-α) gene expression and serum level. Methods We evaluated the serum levels of vitamin D 25(OH)D3 and IFN-α by enzyme-linked immunosorbent assay (ELISA). IFN-α gene expression was measured by real-time polymerase chain reaction (PCR) assay in 123 Egyptian female patients with SLE and in 100 females as a healthy control group. Results Vitamin D deficiency was prevalent in 20.30%, while insufficiency was prevalent in 42.40% of the total group of patients. Serum levels of 25(OH)D3 were significantly decreased in the group of severe disease, and in the group of patients with lupus nephritis. 25(OH)D3 showed highly significant negative correlation with the SLE Disease Activity Index (SLEDAI) in the high activity group and lupus nephritis group. There was a significant negative correlation between 25(OH)D3 and IFN-α serum level and gene expression in all patients; more significant in the group with lupus nephritis. Conclusions The deficiency of 25(OH)D3 has a direct relationship with increase disease activity and nephritis in Egyptian SLE patients, suggesting the need for vitamin D supplementation in these patients. Also, it is directly correlated with increased secretion and gene expression of IFN-α, suggesting its role in pathogenesis of lupus nephritis, to be confirmed by further longitudinal observational studies.

Tumor Necrosis Factor-α, Interleukins-12(p40), 6, and 10 levels in cerebrospinal fluid and outcome prediction in Ossimi sheep with encephalitic listeriosis.

Encephalitic listeriosis in sheep is a life-threatening disease. However, little is known about the cytokine response and their predictive value in this disease. The aim of present study was to assess the prognostic significance of Tumor Necrosis Factor-α (TNF-α), Interleukin-12(p40) (IL-12 p40), Interleukin-6 (IL-6), and Interleukin 10 (IL-10) levels in cerebrospinal fluid (CSF) in sheep with encephalitic listeriosis. Fifty-nine ewes in 14 flocks were diagnosed clinically as having listeriosis. CSF was collected and subjected to bacteriological examination and estimation of selected cytokines. Twenty-eight ewes were confirmed to be infected with Listeria monocytogenes. Based on antimicrobial sensitivity test, sheep were treated and the outcome was recorded as survivors (n=10) and non-survivors (n=18). Cutoff points for CSF cytokines were determined by Receiver operating characteristic analysis (ROC). Association between levels of CSF cytokines and outcome of listeriosis was assessed by logistic regression. TNF-α, IL-6 and IL-12(p40) levels as well as TNF-α/IL-10 ratio were significantly higher in non-survivors than survivors (p=0.002, 0.0021, 0.0033, and 0.001, respectively). However, IL-10 level was significantly lower in non-survivors than survivors (p=0.0058). ROC analysis revealed that IL-6 and TNF-α/IL-10 ratio had the highest AUC values (0.98, 0.984, respectively). Final multivariate logistic regression model showed that TNF-α/IL-10 ratio was the only variable that has predictive value for mortality in diseased sheep (p: 0.001; OR: 7.2; 95% CI: 5.7-9.8). TNF-α showed a positive correlation with IL-12β (r=0.917) and IL-6 (r=0.965). IL-12 (p40) showed also a positive correlation with IL-6 (r=0.906). However, IL-10 showed a negative correlation with TNF-α (r=-0.915), IL-12(p40) (r=-0.790), and IL-6 (r=-0.902). In conclusion, TNF-α/IL-10 ratio may provide predictive information about outcome of encephalitic listeriosis in sheep.