Nermin Raafat

Malathion exposure and insulin resistance among a group of farmers in Al-Sharkia governorate.

OBJECTIVES Exposure to certain environmental toxins may be associated with increased risk of developing diabetes mellitus. The aim of the present study was to investigate the relation between chronic exposure to malathion and insulin resistance among farmers. DESIGN AND METHODS The study included 98 non diabetic farmers who handle agricultural insecticides during their field work. The range of the exposure period for agricultural pesticides was 15-20 years. All farmers were males with mean age 39±12 years. Another 90 administrative employees at Zagazig University Hospitals, non diabetic males age matched were selected as controls. History taking including family history for diabetes, assessment of blood pressure, height, weight, waist circumference and body mass index was done for all participants. Blood samples were withdrawn for measurement of malathion concentration, fasting blood glucose level and fasting insulin level for calculation of homeostasis model assessment of insulin resistance (HOMA-IR). RESULTS 24.5% had positive family history for diabetes. It was observed that there was a significant increase in the mean values of malathion blood concentration among studied farmers compared to corresponding controls. There was a positive correlation between malathion blood concentration, waist circumference and insulin resistance. It was also observed that the increase in the mean values of waist circumference and body mass index was accompanied by a significant increase in the mean values of malathion blood concentration. CONCLUSION The current results suggested that chronic exposure of non diabetic farmers to organophosphorus malathion pesticides may induce insulin resistance. This effect tended to strengthen as waist circumference increases.

Mesenchymal stem cells: In vivo therapeutic application ameliorates carbon tetrachloride induced liver fibrosis in rats.

BACKGROUND Egypt has the highest prevalence of hepatitis C virus in the world with infection rate up to 60%, for which liver fibrosis or hepatic carcinoma is the final outcome. Stem cell therapy provides a new hope for hepatic repair instead of traditional treatment, liver transplantation, as it is safer, gives long term engraftment and avoid expensive immunosuppressive drugs and unexpected hazardous effects. AIM This work aimed at determining the therapeutic potential of mesenchymal stem cells (MSC) in hepatic repair as a new line of therapy for liver fibrosis. METHODS 33 female albino rats were divided into three groups: Group I: 10 rats injected subcutaneously with olive oil, Group II: 13 rats injected with carbon tetrachloride (CCl4) and Group III: 10 rats injected with CCl4 then bone marrow derived MSC from male rats. Blood and liver tissue samples were taken from all rats for biochemical and histological study. RESULTS Liver functions for group II rats showed significant deterioration in response to CCl4 in addition to significant histological changes in liver lobules and portal areas. Those parameters tend to be normal in MSC-treated group. Group III rats revealed normalized liver function and histological picture. Meanwhile, most of the pathological lesions were still detected in rats of second group. CONCLUSION Undifferentiated MSCs have the ability to ameliorate CCl4 induced liver injury in albino rats in terms of liver functions and histological features. So, stem cell therapy can be considered clinically to offer a hope for patients suffering from liver fibrosis.

Mesenchymal stem cells and differentiated insulin producing cells are new horizons for pancreatic regeneration in type I diabetes mellitus

BACKGROUND Diabetes mellitus has become the third human killer following cancer and cardiovascular disease. Millions of patients, often children, suffer from type 1 diabetes (T1D). Stem cells created hopes to regenerate damaged body tissues and restore their function. AIM This work aimed at clarifying and comparing the therapeutic potential of differentiated and non-differentiated mesenchymal stem cells (MSCs) as a new line of therapy for T1D. METHODS 40 Female albino rats divided into group I (control): 10 rats and group II (diabetic), III and IV, 10 rats in each, were injected with streptozotocin (50mg/kg body weight). Group III (MSCs) were transplanted with bone marrow derived MSCs from male rats and group IV (IPCs) with differentiated insulin producing cells. Blood and pancreatic tissue samples were taken from all rats for biochemical and histological studies. RESULTS MSCs reduced hyperglycemia in diabetic rats on day 15 while IPCs normalizes blood glucose level on day 7. Histological and morphometric analysis of pancreas of experimental diabetic rats showed improvement in MSCs-treated group but in IPCs-treated group, β-cells insulin immunoreactions were obviously returned to normal, with normal distribution of β-cells in the center and other cells at the periphery. Meanwhile, most of the pathological lesions were still detected in diabetic rats. CONCLUSION MSCs transplantation can reduce blood glucose level in recipient diabetic rats. IPCs initiate endogenous pancreatic regeneration by neogenesis of islets. IPCs are better than MSCs in regeneration of β-cells. So, IPCs therapy can be considered clinically to offer a hope for patients suffering from T1D.

Vitamin D, Parathormone and Associated Minerals among Students in Zagazig District, Sharkia Governorate, Egypt.

AIM To determine the prevalence of vitamin D deficiency and associated factors among students of age 13-18 years. METHODS DESIGN Community-based cross sectional survey. SETTING Two schools were selected using multistage sampling techniques. SAMPLING Cluster sampling of all enrolled students (550 students). OUTCOME MEASURES Serum levels of 25-hydroxyvitamin D (25 OHD), parathyroid hormone and calcium. Data was collected about nutritional intake, physical activity and lifestyle variables that are potential risk factors for hypovitaminosis D. RESULTS Hypovitaminosis D prevalence was 23.8%, of which 5.3% was deficiency and 18.5% insufficiency. Serum 25 OHD levels inversely correlated with parathyroid hormone levels (r=-0.206, P= 0.00). Low calcium and ionized calcium levels were 40.6% and 45.9%, respectively, and significantly correlated with vitamin D levels. Female students have significantly higher levels of hypovitaminosis D compared to males (29.3% to 15.0%, respectively) and the level of vitamin D significantly improved with increased age. Exposure to sun had a significant effect on vitamin D levels, and physical activity, soft drink consumption and smoking did not. Multinomial regression analysis revealed that age, sun exposure and Ca level were the only significant independent predictors of hypovitaminosis D among the studied group. CONCLUSION Our findings revealed that hypovitaminosis D is a prevalent health problem in adolescents, especially girls, who were at higher risk, and increased age and sun exposure improved vitamin D status among the studied group. There is therefore a need to consider vitamin D supplementation for school children together with increased awareness through a health education program.

Study of non-organ-specific antibodies in children with Genotype 4 chronic Hepatitis C

Background/Aim: Adult studies established a relationship between hepatitis C virus (HCV) infection and the presence of non–organ-specific antibodies (NOSAs). Most studies were carried out on genotypes 1 and 2. Only a few studies addressed that issue in pediatrics. No studies have been carried out on autoimmunity and genotype 4 in children. We aim to investigate NOSAs in 80 Egyptian children with chronic HCV infection along with studying the underlying genotype of HCV, and correlating autoimmunity with the epidemiological, clinical, biochemical, and virological features. Materials and Methods: HCV-RNA was assayed by the polymerase chain reaction and viral genotypes were determined. NOSAs were measured and liver biopsies were taken for histopathological examination. Results: Genotype4 was the only detected genotype in the included 80 patients. Anti-smooth muscle antibodies (ASMA) were the only detected antibodies in 32 (40%) patients, always with V specificity (vessels only) at titers ranging from 1:20 and 1:160. Anti-nuclear antibodies (ANA) and liver–kidney microsomal antibodies-1 (LKMA-1) were not detected in any of our patients. Epidemiologic and clinical features did not significantly differ between autoantibody-positive and -negative patients. Among biochemical features, significantly high levels of total bilirubin, albumin, immunoglobulins, alkaline phosphatase, and gamma-glutamyl transpeptidase were found in the antibody-positive group. Conclusion: Genotype 4 HCV is the prevailing genotype in Egyptian children with chronic HCV infection. A consistent proportion of these children with chronic HCV infection circulate non–organ-specific autoantibodies. The prevalence of ASMA and the absence of ANA and LKMA-1 might be related to the unique situation in Egypt with unique prevalence of genotype 4. More studies are warranted on larger pediatric population to validate these findings.

Assessment of neutralizing interleukin-4 effect on CD133 gene expression in colon cancer cell line

&NA; Colorectal cancer may be maintained by cancer stem‐like cells (CSCs) that express the cell surface marker CD133. CSCs (CD133+ cells) exhibits greater resistance to the chemotherapy and this resistance may be mediated in part by an autocrine response to IL4. The aim of the study was to assess the effect of anti‐IL4 antibody alone or in combination with chemotherapy on the CD133 expression andthe tumor growth. We used Caco cell line in our experiments and the samples were as the following; untreated colorectal cell line, cells treated by chemotherapy, cells treated by anti‐IL4 antibody in 3doses (2.5, 5, 10 &mgr;g/ml), cells treatedby combination of chemotherapy and anti‐IL4 antibody in 3 doses. Results of our in vitro studies demonstrated that anti‐IL4 inhibited growth of Caco cell line in a dose‐dependent manner revealing a 32.11% inhibition at the highest concentration (10 &mgr;g/ml). There was further significant inhibition by combination of anti IL4 and chemotherapy in a dose response manner when compared to group treated by chemotherapy only. These effects were associated with decreased expression of CD133 in tumor cells also. Lastly, anti‐IL4 antibody stimulated apoptosis. Our study suggested that neutralizing of IL4 by anti IL4 antibody affect the CD133+ cells may be by increasing their apoptosis. The effects of anti IL4 antibody either, alone or in combination with chemotherapy, inhibited the tumor growth and decreased the viable tumor cells. Furthermore, neutralizing of IL4 increased the efficacy of chemotherapy treatment.

Abstract 2490: CD40 ligand expressing recombinant vaccinia virus (rVV40L) modulation of central memory CD8-mediated immune response

In cancer immunotherapy, induction of tumor-reactive CD8+ T cells displaying phenotypic and functional profile of central memory T cells (TCM) is associated with favorable prognosis. A key element in the generation of TCM CD8+ cells is represented by the help provided by CD4+ T cells during the priming of naive CD8+ T cells. In particular CD40 ligand (CD40L) expressed and/or secreted by activated CD4+ T cells triggers CD40 receptor expressed on antigen presenting cells (APCs) thereby enhancing their antigen presentation capacity. In order to bypass the requirement of CD4+ T cells we generated a non-replicating recombinant vaccinia virus encoding human CD40L (rVV40L) and compared its ability to shape CD8-mediated immune response to soluble CD40L recombinant protein (sCD40L). In this regard, our data clearly underline the different biological properties of membrane-bound CD40L, as provided by rVV40L infection, as compared to its soluble form, in CD14+ APCs activation. Notably, considering expression of IL-12p40, IFN-a and -b genes, cytokines of proven relevance for memory T cell induction, rVV40L-infection was much more potent than s40L-treatment alone or combine with WT infection. In parallel, s40L-stimulation induced a much more significant expression of IL-10 and indoleamine-2, 3-dioxygenase (IDO) genes encoding immunosuppressive factors. Considering a panel of molecules involved in the generation of the immunological synapse with T cells on CD14+ cells, rVV40L appeared to promote a less intense up regulation of CD80 co-stimulatory molecules but, most importantly, only a minor increase of programmed death ligand-1 (PD-L1). Therefore, gene expression and phenotypic profiles suggested that rVV40L-infected CD14+ cells might be highly effective APC in the induction of TCM CD8+ cells. Indeed, a single in vitro stimulation of naive CD8+ T cells by rVV40L-infected CD14+ cells, in the absence of CD4+ T cells, was able to promote the rapid generation of central memory TAA (MAGE and MART-1) specific CD8+ T cells. These TCM were characterized by the typical central memory phenotype, as indicated by co-expression of CD45RO, CD62L and IL-7Ra, and by the high proliferative potential upon antigen recognition. Collectively our data indicate that rVV40L efficiently modulates the quality of different APC signals delivered during the formation of the immunological synapse with CD8+ T cells. These in-vitro observations validate the strong clinical potential of our recombinant vaccinia virus constructs co-expressing CD40L for cancer immunotherapies. Citation Format: Emanuele Trella, Evangelos Panopoulos, Nermin Raafat, Chantal Mengus, Emmanuel Traunecker, Swantje Heidtmann, Michael Heberer, Daniel Oertli, Giulio Cesare Spagnoli, Paul Zajac. CD40 ligand expressing recombinant vaccinia virus (rVV40L) modulation of central memory CD8-mediated immune response. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2490. doi:10.1158/1538-7445.AM2015-2490

EARLY POSTOPERATIVE MEASUREMENT OF CALCIUM AND PARATHRMONE LEVELS AS PREDICTORS OF EARLY PARATHYROID INSUFFICIENCY AFTER TOTAL THYROIDECTOMY

Background: Declines in serum calcium or intact parathyroid hormone (iPTH) levels after surgery have been suggested as being reliable predictors of postoperative hypocalcemia. Although measurements of serum calcium or iPTH allow the identification of patients who have no risk of hypocalcemia after total thyroidectomy, it is not always easy to predict which patients can be discharged early from the hospital or to identify those requiring close monitoring of serum calcium levels or those that should receive calcium and vitamin D supplements. Aim of the work To evaluate early postoperative parathyroid insufficiency following total thyroidectomy by estimating parathyroid hormone and calcium level in the early postoperative period, and whether we can rely on to start Ca supplement or not. Patients and methods: This study was done in General Surgery department, Zagazig University on 50 patients; 10 males (20%)) 40 females (80%) who underwent total thyroidectomy. Serum total Calcium and Parathormone levels were evaluated in all patients 6 h and 24 h in early postoperative period. Results: Our results showed highly significant VALUE OF relative decline of serum calcium and parathyroid hormone 6 h and 24 h after total thyroidectomy. The results showed also highly significant positive correlation between serum calcium and parathyroid hormone both preoperative and postoperative and highly significant positive correlation between relative decline in serum calcium and relative decline of serum parathyroid hormone at both 6h and 24 h after total thyroidectomy IN HYPO CALCEMIC PATIENTS. There was no significant relationship between hypocalcemia and any of demographic, clinical, ultrasonographic and pathological data. There is significant relationship between both relative decline of serum calcium and relative decline parathyroid hormone and postoperative hypocalcemia. Conclusion: Relative decline of parathyroid hormone 6 h and 24 h after total thyroidecomy is a good predictor of postoperative hypocalcemia and is reliable for allowing safety discharge of patients 24 h after total thyroidectomy. Key words: Total thyroidectomy, Calcium, Parathyroid hormone

Study of non-organ-specific antibodies in Egyptian children with genotype-4 chronic hepatitis C

Background/rationale Adult studies established a relationship between hepatitis C virus (HCV) infection and the presence of anti-nuclear antibodies (ANA), anti-smooth-muscle antibodies (ASMA), and anti-liver–kidney-microsomal type 1 antibodies (LKMA-1). No studies have been carried out on autoimmunity and genotype-4 in children. We aimed to investigate non-organ-specific autoantibodies in Egyptian children with chronic HCV infection and to correlate autoimmunity with epidemiological, clinical, biochemical, and virological features. Methods All enrolled patients (n=80) were positive for anti-HCV antibodies. HCV-RNA was assayed by PCR and viral genotypes were determined. ANA, ASMA, and LKMA-1 levels were assessed and liver biopsies were taken for histopathological examination. Results Genotype 4 was the only detected genotype in our cohort. ASMA were the only detected autoantibodies; they were detected in 32 patients (40%), always with V specificity (vessels only) at titers ranging from 1 : 20 to 1 : 160. ANA and LKMA-1 were not detected in any of our patients. Epidemiologic and clinical features did not significantly differ between autoantibody-positive and autoantibody-negative patients. Among biochemical features, significantly high levels of total bilirubin, albumin, immunoglobulins, alkaline phosphatase, and &ggr;-glutamyl transpeptidase were found in the antibody-positive group. Conclusion Genotype-4 HCV is the prevailing genotype in Egyptian children with chronic HCV infection. A consistent proportion of these children with chronic HCV infection have non-organ-specific autoantibodies in their circulatory system. The prevalence of ASMA and the absence of ANA and LKMA-1 might be related to the unique situation in Egypt of unique prevalence of genotype 4. More studies are warranted in larger pediatric populations to validate these findings.

Abstract 2228: Folate-related genes polymorphism and risk of pediatric acute lymphoplastic leukemia

Background: Childhood acute lymphoblastic leukemia (ALL) is the most common malignancy affecting children, constituting about 30% of all cancers among children. It constitutes about 75% of pediatric acute leukemia with peak incidence between ages 3 and 4. The assertion that ALL may have a genetic basis has long been pursued through association studies based on candidate genes as folate metabolism. Polymorphisms in folate pathway genes may influence the susceptibility to acute lymphoblastic leukemia. Increased risk of ALL was observed in reduced folate carrier protein (RFC1) 80GA variant carriers. A 1.4-fold reduction in ALL risk was observed for carriers of the methyl tetrahydrofolate reductase (MTHFR) 677T allele. Objectives: this study was carried out to evaluate the contribution of MTHFR C677T and RFC1 80GA gene polymorphism and susciptability of childhood acute lymphoblastic leukemia. Materials M 95% CI, 0.9-1.5; P Citation Format: Nermin Raafat, Amal Gharib, Usama Elsafy, Tamer Hassan. Folate-related genes polymorphism and risk of pediatric acute lymphoplastic leukemia. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2228. doi:10.1158/1538-7445.AM2014-2228