Autophagy - related 16 - like 1 single nucleotide gene polymorphism increases the risk and severity of Cryptosporidium parvum infection.

Cryptosporidium parvum (C. parvum) is an intracellular parasite of the intestinal cells. It causes cryptosporidiosis that can be fatal in immunosuppressed individuals. Autophagy is a process to eliminate intracellular microbes. The autophagy-related 16 - like 1 (ATG16L1) gene encodes proteins involved in the autophagy pathway. Single nucleotide polymorphism (SNP) in this gene increases the invasion and survival of the intracellular microbes. This study aimed to assess whether SNP in the ATG16L1 gene influences the risk and severity of cryptosporidiosis. Group I: cases with C. parvum infection (C. parvum, n = 40) and group II: healthy control (HC, n = 120) were included. Genotyping of the ATG16L1 gene was done for all participants to determine the polymorphism status as AA, GG, or AG genotype. A significant association between C. parvum infection and ATG16L1 genotypes was detected. C. parvum group had a significantly higher frequency of GG genotype and G allele when compared to HC group. The genotypes (AG + GG) and G allele had 2.428 and 2.13 folds risk of C. parvum infection when compared to the AA genotype and the A allele. Patients with the AG + GG genotype had statistically significant higher Cryptosporidium oocyst counts in stool, higher infection intensity, more frequency of vomiting and dehydration, longer disease duration, and more recurrence. The GG or AG genotypes were independent risk factors in the disease severity (p- value = 0.013). In conclusion, ATG16L1 SNP increased the risk and severity of cryptosporidiosis. Thus, individuals with such SNP can benefit from autophagy up-regulating approaches in decreasing the risk and controlling C. parvum infection.

CMV, B and C hepatitis among multi-transfused hereditary hemolytic Anemia children: an updated Egyptian experience.

BACKGROUND AND OBJECTIVES: Regular blood transfusion has improved the overall survival and quality of life for patients with hereditary hemolytic anemias. Nevertheless, it carries a real risk of acquisition of blood-borne virus infections, especially viral hepatitis. The purpose of the current study is to present an Egyptian update on blood-borne hepatitis C & B viruses (HCV & HBV) and cytomegalovirus (CMV) among multi-transfused Egyptian children with hereditary hemolytic anemias, especially after implementation of national preventive programs in Egypt. PATIENTS AND METHODS: All pediatric patients with hereditary hemolytic anemias who have regular follow-up and received frequent blood transfusion at the Pediatric Hematology Units, Menuofia and Zagazig Universities Hospitals, Egypt, during the study period, were recruited. They were tested for hepatitis B surface antigen (HBVsAg), hepatitis C antibody (HCVab), and CMV immunoglobulin M (IgM) serology. Those with positive results were confirmed by real-time polymerase chain reaction (PCR). RESULTS: Four hundred and seventy-seven hereditary hemolytic anemia patients fulfilled the study inclusion criteria. Their ages ranged from 2 to 18 years, 54.9% of them were males. Seroprevalence of HCVab and CMV-IgM were (14.7% & 6.7% respectively) and they were confirmed by PCR. None of the studied cases were HBVsAg positive. Seropositivity for HCV was significantly associated with older age of the patients, higher transfusion frequency, longer disease duration, and higher mean serum ferritin. CONCLUSION: HCV followed by CMV infections still represent a significant problem for patients with hereditary hemolytic anemias. Nationwide plans should be taken to ensure meticulous and highly sensitive methods of blood screening before transfusion. On the other hand, it seems that HBV compulsory vaccination had succeeded to eliminate HBV infection.

Platelet indices as an assessment tool of septic acute kidney injury.

Platelet (PLT), one of blood cells, plays a major role in physiological and pathological processes such as coagulation, thrombosis, inflammation, and keeping the integrity of vascular endothelium. There are a group of parameters that are used to measure the total amount of PLTs, PLTs morphology, and proliferation. PLT indices are associated with the severity of illness and patients' prognosis. It was reported that mean platelet volume (MPV) was raising synchronously with interleukin (IL)-6 and C-reactive protein in sepsis, and was correlated to the severity of the disease. We aimed to study PLT indices and its changes in sepsis and septic acute kidney injury (AKI) patients to assess the disease and its severity. The present study is a cross-sectional study, had been carried out at Menoufia University hospitals from August 2017 to August 2019. The various platelet indices [MPV, platelet distribution width (PDW) and plateletcrit (PCT)] are considered as outcome variables were compared among controls, cases with sepsis, and cases with sepsis associated AKI. Group I (31) cases with the clinical diagnosis of septic AKI, Group II (33) cases with the diagnosis of sepsis, and Group III (28) consecutive persons marked as negative in the output of the cell counter were taken as controls. Data were tabulated and statistically analyzed. There were 15 men and 15 women for Group I (septic AKI), 17 males and 16 females for Group II (sepsis) and 15 men and 13 women healthy controls as a control group. According to PLT indices MPV, there was a significant statistical difference (P1 <0.01) between Group I and II of patients as it were12.06 ± 1.23, 11.01 ± 1.20, respectively, and PDW also there was a significant statistical difference (P1 <0.01) as it were16.01 ± 2.33, 13.97 ± 2.14, respectively, and PCT there was no significant difference between the two groups. Furthermore, there was a significant statistical difference between Group I and II of patients according to procalcitonin, TNF-α and IL-10. From these results, we conclude that there were a statistical significant difference between the patient groups of critically ill.

Deregulated Expression of Long Non-coding RNA HOX Transcript Antisense RNA (HOTAIR) in Egyptian Patients with Multiple Myeloma.

Increasing evidence of involvement of non-coding RNAs, especially long non-coding RNAs (lncRNAs), in the molecular biology of various malignancies have been recently reported. Their utilization as markers for diagnosis, prognosis and evaluation of treatment response was widely investigated. As the impact of lncRNA HOTAIR on multiple myeloma (MM) was not properly highlighted, we aimed to explore the expression levels of HOTAIR in three groups of MM patients and to analyze its relationship to different patients' characteristics. Plasma samples were withdrawn from 24 newly diagnosed MM patients, 23 post-therapy patients in complete response (CR) or very good partial response (VGPR) and 15 patients who had either progressive disease (PD) or relapse. The expression of lncRNA HOTAIR in MM patients and 20 healthy controls was analyzed by quantitative reverse transcription polymerase chain reactions. HOTAIR was significantly upregulated in newly diagnosed and PD/relapse categories in comparison with controls and MM patients who had achieved CR or VGPR (P < 0.001). Furthermore; HOTAIR expression levels correlated with the percentage of malignant plasma cells in bone marrow (P = 0.006) and disease stage (ISS stage) (P = 0.031). HOTAIR may be employed as  prognostic molecular marker and novel therapeutic tool for newly diagnosed MM patients.

XRCC1 Gene Polymorphism Increases the Risk of Hepatocellular Carcinoma in Egyptian Population.

Several major risk factors for hepatocellular carcinoma (HCC) have been identified, including chronic infection of hepatitis B virus (HBV) and hepatitis C virus (HCV). Nevertheless, only a fraction of infected patients develops HCC during their lifetime suggesting that genetic factors might modulate HCC development. X-ray repair cross complementing group1 (XRCC1) participates in the repair pathways of DNA. AIM: to investigate the association between XRCC1 gene polymorphism and HCC in Egyptian chronic hepatitis C patients. METHODS: This study was assessed on 40 patients with HCC secondary to chronic HCV infection who were compared to 20 cirrhotic HCV patients and 40- age and gender- matched healthy control group. After collection of relevant clinical data and basic laboratory tests, c.1517G>C SNP of XRCC1 gene polymorphism was performed by (PCR-RFLP) technique. RESULTS: A statistically higher frequency of XRCC1 (CC, GC) genotypes and increased (C) allele frequency in patients with HCC was found in comparison to cirrhotic HCV patients as well as control group. In addition, patients with the XRCC1 (CC, GC) genotypes had significantly higher number and larger size of tumor foci and significantly higher Child Pugh grades. Multivariate analysis showed that the presence of c.1517G>C SNP of XRCC1 gene is an independent risk for the development of HCC in chronic HCV patients with 3.7 fold increased risk of HCC development. IN CONCLUSION: XRCC1 gene polymorphism could be associated with increased risk of HCC development in chronic HCV Egyptian patients.
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IL-10 polymorphisms and T-cell subsets could affect the clinical presentation and outcome of childhood immune thrombocytopenia in Egyptian population.

The aim is to study IL-10 polymorphisms and IL-10 level and assess their relation to T-cell subsets in childhood immune thrombocytopenia (ITP). In all, 40 (25 acute, 15 chronic) ITP child patients were investigated at time of presentation, compared to 15 healthy, age- and gender-matched controls and followed up for 1 year to determine chronic cases. Studying the effect of IL-10 promoter polymorphism was done by PCR-RFLP, IL-10 level was determined by ELISA, natural killer cells and T-cell subsets were evaluated by flow cytometry. Subjects with IL-10 promoter (1082 AA and 592 AA) genotypes had lower IL-10 levels and had lower CD4%, higher CD8%, lower CD4/CD8 ratio and lower T-reg%. IL-10 polymorphisms had no effect on NK%. IL-10 serum levels and IL-10 promoter polymorphic genotype frequencies are not different between ITP cases and controls; however, in ITP patients, IL-10 promoter (1082 AA and 592 AA) genotypes and associated lower CD4, higher CD8, lower CD4/CD8 ratio is associated with more severe thrombocytopenia at presentation and had a poorer response to first-line treatment. Patients with lower T-reg cells had a higher tendency to develop chronic ITP. IL-10 level and polymorphisms as well as disturbed T-cell subsets percentages are demonstrable effectors of immune dysfunction in ITP and can affect the presentation and outcome of childhood ITP.

Toxoplasma gondii as a possible causative pathogen of type-1 diabetes mellitus: Evidence from case-control and experimental studies.

Toxoplasma gondii is the causative parasite of an important worldwide disease. This obligate intracellular parasite can infect and replicate inside any nucleated cells including those of pancreas. Insulin is a hormone secreted by the pancreas and is responsible for controlling blood glucose concentration. Deficiency of insulin production accounts for the occurrence of type-1 diabetes mellitus (T1D). Thus, theoretically, toxoplasmosis could play a possible role in the development of T1D. However, the studies on this theory are still insufficient; therefore, this work was designed. Interestingly, in the case-control study, seropositivity of anti-Toxoplasma IgG was significantly higher among T1D (86.37%) in comparison with T2D (66.67%) and the control group (60%). Moreover, the odd ratio of chronic toxoplasmosis was 4.2 folds higher among T1D patients than among controls. The experimental study included acute and chronic Me49 T. gondii infected mice groups in addition to a control group. Pathological examination revealed the presence of T. gondii zoites adjacent to the islets of Langerhans and in pancreatic parenchyma of acutely infected mice. With chronic infection, there was a significant reduction of islets number and sizes in association with grade-1 insulitis. Additionally, the immunohistochemical study showed significant infiltration of the islets of chronically infected mice by CD8+ and CD45+ immune cells. In contrary to the control group, the islets of the chronic group showed significantly higher expression of the apoptotic marker caspase-3 and a significantly lower expression of the proliferation marker Ki69. Finally, a significant reduction of insulin expression in the islets of chronic infection group was detected in association with a significant increase in serum glucose concentrations; however, the establishment of diabetes did not occur throughout this work. Thus, this study presents an evidence for the probable role of chronic toxoplasmosis in the development of T1D which should be considered in further studies.

Utility of serum resistin in the diagnosis of neonatal sepsis and prediction of disease severity in term and late preterm infants.

Introduction Resistin is a proinflammatory hormone recently proposed as a sepsis biomarker. Our aim was to evaluate the diagnostic and prognostic values of this marker in neonatal sepsis. Methods This is a prospective observational study that includes 60 term and late preterm neonates with proven and possible sepsis besides 30 healthy controls. Resistin and other biomarkers, like C-reactive protein (CRP), were measured within 2 h of neonatal intensive care unit (NICU) admission. Infants were monitored and the primary outcome was 30-day mortality. Results Resistin was higher among septic neonates compared with controls (P<0.001). Resistin had an area under the receiver operating characteristic (ROC) curve of 0.994 for differentiating septic infants from controls. The area under the curve (AUC) for differentiating infants with culture-proven sepsis from controls was 0.999 compared with an AUC of 1 for CRP. The other markers, like platelet count, were inferior to resistin and CRP. Resistin was positively correlated with CRP [Spearman's correlation coefficient (rs)=0.55, P<0.001]. No significant differences in resistin levels were noted between survivors and non-survivors but resistin was higher among infants with severe sepsis (P=0.015) and among those who needed mechanical ventilation (P<0.001). Conclusion Resistin is useful for the diagnosis of neonatal sepsis. Resistin failed to predict mortality but was associated with indicators of disease severity.

Serum stromal-derived-factor-1 (CXCL12) and its alpha chemokine receptor (CXCR4) as biomarkers in neonatal sepsis.

BACKGROUND: Neonatal sepsis remains one of the leading causes of morbidity and mortality both among term and preterm infants. Advances in neonatal care improved survival and reduced complications in preterm infants. Chemokines are chemotactic cytokines that give directional guidance for leukocyte migration during inflammatory process. The chemokine CXCL12 and its receptor CXCR4 are now known to play an important role in inflammatory states. However, its value as a biomarker in neonatal sepsis is unclear. OBJECTIVES: To assess the value of measuring the serum levels of alpha-chemokine receptor type 4 (CXCR-4) and stromal-derived-factor-1 (CXCL12) in diagnosis of late onset neonatal sepsis. SUBJECT AND METHODS: Serum levels of CXCL12 and CXCR4 were determined in 38 full term neonates, 23 cases of late onset sepsis (13 males and 10 female), and 15 healthy neonates as control (six males and nine females) by ELISA technique and flow-cytometry. RESULTS: Serum levels of CXCR4 and CXCL12 were significantly higher in neonates with late onset sepsis compared with the non-septic ones. The sensitivity, the specificity, and the overall accuracy of CXCL12 were 100%. The sensitivity of CXCR4 was 87%; the specificity was 80% and the overall accuracy was 84%. CONCLUSIONS: Serum CXCR4 and CXCL12 levels increase significantly in septic neonates and they are valuable marker in diagnosis of neonatal sepsis. Serum concentrations of both chemokines represent promising novel biomarkers for neonatal sepsis.

Study of CD4+, CD8+, and natural killer cells (CD16+, CD56+) in children with immune thrombocytopenic purpura.

OBJECTIVE/BACKGROUND: To assess the percentage of CD4+, CD8+, and natural killer cells (CD16+, CD56+) in children with immune thrombocytopenic purpura (ITP) at presentation and study their impact on disease chronicity. METHODS: This case-control study was conducted at the Pediatric Hematology and Oncology Unit, Menoufia University Hospital (tertiary care center in Egypt). The study was held on 30 children presenting with ITP; they were followed-up and classified into two groups: 15 children with acute ITP; and 15 children with chronic ITP. Patients were compared to a group of 15 healthy children of matched age and sex. Measurements of CD4+, CD8+, and natural killer cells (CD16+, CD56+) by flow cytometry were assessed and compared in these groups. RESULTS: CD4+ and CD4+/CD8+ were significantly lower in acute and chronic patients than the control group (p<0.05 and p<0.001, respectively), with no significant difference between acute and chronic patients (p>0.05). However, CD8+ was significantly higher in acute and chronic patients than the control group (p<0.05), with no significant difference between acute and chronic patients (p>0.05). Natural killer cell percent was significantly lower in acute patients than the control group (p<0.001), with no significant difference between chronic and control groups (p>0.05). CONCLUSION: ITP is associated with immunity dysfunction denoted by the increase in cytotoxic T lymphocytes and the decrease in natural killer cells.

Effect of Phototherapy on CD4, CD8 and Natural Killer Cells of Full Term Neonates with Indirect Hyperbilirubinemia.

Phototherapy is generally considered a very safe and well-tolerated treatment for hyperbilirubinaemia. However, clinical users should be aware of the unwanted effects of using phototherapy. Affection of neonatal immune system due to phototherapy has been reported. This study aimed to evaluate the effect of phototherapy on level of CD4+, CD8+ and natural killer (NK) (CD16+ & CD65+) lymphocytes subsets in neonates. The number of these lymphocytes was measured 72 hrs after phototherapy exposure in 30 full term neonates with indirect hyperbilirubinemia and compared to those of 25 healthy controls using flow cytometry. Results showed non-significant changes of the tested lymphocyte subsets after 72 hrs exposure to phototherapy. In conclusion, phototherapy has no significant effect on the level of circulating CD4+, CD8+ and NK lymphocytes.