Assessment of the natural killer cell, IFN- and IL-10 blood levels in children with attention-deficit/hyperactivity disorder.

Attention-deficit/hyperactivity disorder (ADHD) is a neurodevelopmental disorder, affects millions of youngsters and typically persists into adulthood. The pathophysiology of ADHD may be due to an impaired immune response, common genetics, and environmental linkages, as all have been suggested as potential underlying mechanisms for ADHD. During systemic inflammation, natural killer (NK) cells can produce pro-inflammatory cytokines like interferon (IFN- ) and anti-inflammatory cytokines like interleukin (IL-10); this demonstrates the importance of both of their roles as regulators to counteract inflammation and prevent immune-mediated host damage. In this work we aimed to determine the role of inflammation in children with ADHD by measuring the level of NK cells in peripheral blood compared to typically developing children besides estimating the inflammatory cytokines INF- and IL -10 in both groups. This study included 50 children diagnosed with ADHD based on the Diagnostic and Statistical Manual of Mental Disorders-5th edition and 50 age and sex- matched normally developed children, as controls. The estimation of NK was done using flow cytometry, while the studied cytokines were measured using the ELISA technique. We found that children with ADHD had a significantly higher level of NK cells in peripheral blood compared to controls (p < 0.001). Furthermore, increased IFN -, while decreased IL-10 serum levels were observed in children with ADHD compared to their control group. In conclusion our findings suggested that children with ADHD may have impaired immune responses, as NK cells were increased in peripheral blood compared to the control group. Also, the serum level of IFN - was higher, while the serum level of IL-10 was lower in ADHD children as compared to controls.

Serum metabolomic profiles and semaphorin-3A as biomarkers of diabetic retinopathy progression.

Diabetic retinopathy (DR) is a typical microvascular complication of diabetes mellitus (DM) and it remains one of the leading causes of vision loss worldwide. Studies postulated that a distinct metabolic signature of DR exists and can be resolved from that of diabetes alone. Serum Semaphorin3A (Sema3A) levels have also been found to be correlated with the phenotypes of diabetic retinopathy. We aimed to analyze and identify serum metabolites and serum Sema3A levels that could be useful biomarkers of DR progression. This cross-sectional study included 45 type 2 diabetes (T2D) patients. Diabetic patients were divided into three groups based on the status of their complications: non-DR (NDR, n=15), non-proliferative DR (NPDR, n=15), and proliferative DR (PDR, n=15) groups. Serum metabolomic profiles of these patients were determined by using high-performance liquid chromatography-mass spectrometry (HPLC-MS), and serum Sema3A levels measured by ELISA. Metabolomics analysis revealed a set of metabolites that were altered in the serum of PDR patients as compared with NPDR and NDR groups. Among these metabolites total asymmetric dimethylarginine (ADMA) and Kynurenine were potential predictors of PDR patients. Significantly higher serum levels of Sema3A in PDR patients as compared with NPDR and NDR groups (p < 0.001), their serum levels were positively correlated with the central macular thickness (r= 0.952, p < 0.001) and negatively correlated with the superficial macular density (r=-0.952, p < 0.001). In conclusion, the metabolite signatures identified in this study and serum Sema3A levels could be proposed as biomarkers for DR development and progression in T2D patients. However, Sema3A was superior to metabolomics in the prediction of the severity of DR.

Toxoplasmosis and cytomegalovirus infection and their role in Egyptian autistic children.

Autism is a neurodevelopmental disorder with a significantly increased incidence rate across the world over the past few years. Toxoplasmosis and cytomegalovirus (CMV) infection are globally prevalent and have been associated with diverse neurological and psychiatric disorders. A few studies have demonstrated the role of toxoplasmosis and CMV as potential etiological factors for autism. Accordingly, this study was performed to estimate the relationship between toxoplasmosis and CMV infection in children with autism as well as to assess their impact on the Childhood Autism Rating Scale (CARS) score. A total of 45 autistic children (6 girls, 39 boys) and 45 (21 girls, 24 boys) healthy control children were enrolled in our study. Their blood samples were collected and tested for the presence of Toxoplasma and CMV (IgG and IgM) antibodies and DNA by ELISA and real-time PCR (RT-PCR), respectively. Toxoplasmosis was detected in 11 (24.4%) autistic children through the ELISA [10 (22.2%) IgG + /IgM - and 1 (2.2%) IgG + /IgM +]; however, RT-PCR assay recorded only 1 positive case (2.2%), while it was detected in 10 (22.2%) control children through ELISA [9 (20%) IgG + /IgM - and 1 (2.2%) IgG + /IgM +] and 1 (2.2%) by RT-PCR. On the other hand, CMV infection was detected in all autistic children with 44 (97.8%) testing positive by ELISA [24 (53.3%) IgG + /IgM - , 18 (40%) IgG + /IgM + and 2 (4.4%) IgG - /IgM +] and 25 (55.6%) testing positive by RT-PCR assay. In addition, ELISA assay recorded 43 (95.6%) [19 (42.2%) IgG + /IgM + and 22 (48.9%) IgG + /IgM - and 2 (4.4%) IgG-/IgM +] and RT-PCR recorded 21 (46.7%) positive samples in control children with CMV. No significant difference was noted between autistic and control children for the overall prevalence of Toxoplasma or/and CMV infection. Similarly, the CARS score indicated a non-significant difference with Toxoplasma or/and CMV infection. Our data does not show an association between autism and toxoplasmosis or/and CMV infection. Nevertheless, considering that autistic children are at a high risk of contracting these infections, further studies with a larger sample size are recommended.

The Relationship Between FoxP3 and SOCs3 Gene Expressions and Disease Activity in Rheumatoid Arthritis.

BACKGROUND: Immune dysregulation plays an important role in the pathogenesis of rheumatoid arthritis (RA). The CD4+CD25 high FoxP3+ subset of regulatory T cells plays an essential role in preventing autoimmunity and maintaining immune homeostasis. Negative regulation of JAK/STAT signaling is controlled by Suppressor of Cytokine Signaling (SOCs3) proteins. SOCs is produced at lower levels in RA. Our aim was to evaluate the expressional dysregulation of SOCs3 and FoxP3 genes in RA patients in relation to disease activity. METHODS: We have recruited 90 patients with RA and 60 healthy controls in case control study. Whole blood samples were collected from RA patients and healthy subjects. The measurement of FoxP3 and SOCs3 gene expression was performed by real-time PCR (qPCR). RESULTS: Patients with RA had significantly decreased expression levels of FoxP3 and SOCs3 genes in comparison with controls (P<0.001), in addition to the insignificance correlation of both genes with disease activity in RA patients. CONCLUSION: FoxP3 and SOCs3 genes showed significant defects in rheumatoid arthritis patients with no significant difference in disease activity.

Impact of miR-155 rs767649 Polymorphism on Rheumatoid Arthritis Activity in Egyptian Patients.

BACKGROUND: Rheumatoid arthritis (RA) is a chronic inflammatory condition that impacts not only the musculoskeletal system but also various other systems in the body, including the cutaneous, ocular, respiratory, cardiovascular, and circulatory systems. MicroRNAs (miRNAs) are a class of naturally occurring and highly conserved transcripts that primarily function in the regulation of gene expression. They accomplish this by facilitating the degradation of messenger RNA (mRNA) or by repressing mRNA translation. miRNAs are well-known regulators of a variety of cellular processes. Therefore, we aimed to detect the impact of miR-155 rs767649 polymorphism on RA activity. METHODS: This case-control study included 66 Egyptian patients with RA who visited Al-Zhraa University Hospital, Internal Medicine Department, Cairo, Egypt, and 50 apparently healthy control subjects matched for age and sex. The participants were subjected to full clinical evaluation, including assessments of the disease activity score (DAS), erythrocyte sedimentation rate (ESR), liver and kidney function, anti-cyclic citrullinated peptide antibody (anti-CCP), and miR-155 polymorphism using real-time polymerase chain reaction (PCR). RESULTS: In the RA group, the majority (98.5%) were female, with a mean age of 43 years, while in the control group, 94% were female, with a mean age of 43.4 years. Comparison of laboratory parameters indicated significantly lower hemoglobin levels, higher ESR, and higher serum creatinine and anti-CCP levels in the RA group than in the control group. The RA group had a significantly higher frequency of TT genotypes and significantly lower frequencies of TA and TT genotypes than the control group. Considering the TT genotype and T allele as references, TA, AA, and TA/AA genotypes in the dominant model; AA in the recessive model; and A allele were significantly associated with protective effects against RA development (p<0.05, odds ratio<1). CONCLUSION: rs767649, the functional variant of miR-155, plays an important role in susceptibility to the increased risk of RA, suggesting that miR-155 can be used as a therapeutic target for the treatment of Egyptian patients with RA.

Study of transforming growth factor beta 1 gene +869T/C polymorphism in Egyptian rheumatoid arthritis patients and its relation to disease activity.

This study intended to explore the relationship between the +869T/C polymorphism of the transforming growth factor-ß1 (TGF-ß1) gene and rheumatoid arthritis (RA) predisposition and activity in Egyptian patients. The study involved 30 patients suffering from RA and 30 apparently healthy participants as the control group. The +869T/C polymorphism of the TGF-ß1 gene was determined by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) process. The TGF-ß1 + 869 CT genotype and CT+TT genotypes in RA patients showed a significant increase than the control group (OR=3.782 and 3.824, CI=1.046-13.680 and 1.150-12.713, P=0.043 and 0.029, respectively). T allele showed a significant increase in patients than in controls (OR= 2.104, CI 1.015- 4.361, P = 0.046). The TGF-ß1 +869 CT+TT genotypes were accompanied by higher DAS-28 scores which express higher disease activity, and increased levels of RF, Anti-CCP, ESR, and CRP. In conclusion, the TGF-ß1 +869T/C gene polymorphism may be accompanied by an increased predisposition to RA and with its severity in Egyptian RA patients.

Urinary IgG, serum CX3CL1 and miRNA-152-3p: as predictors of nephropathy in Egyptian type 2 diabetic patients.

The purpose of this study was to assess the role of urinary IgG, serum CX3CL1 and miRNA 152-3p levels as predictors of nephropathy in type 2 Egyptian diabetic patients. Sixty type 2 diabetic patients and twenty healthy controls were enrolled in a cross-sectional study. Then they were grouped into: three groups based upon urine albumin excretion (UAE). The expression of miRNA 152-3p in serum was measured using quantitative polymerase chain reaction (RTq-PCR). Serum CX3CL1 and urinary IgG concentrations were measured by ELISA. RTq-PCR revealed that serum miRNA-152-3p levels in patients were significantly higher than in controls. There was significant differences between group with normoalbuminuria and groups with diabetic nephropathy DN as regard to age, duration of nephropathy, Albumin/Creatinine ratio (A/C ratio), creatinine, urine IgG, CX3CL1 and HbA1c. In diabetic patients, there was a significant positive correlation between miRNA-152-3p levels and disease duration only as well as significant positive correlations between urinary IgG levels and age, disease duration, serum creatinine, A/C ratio, and urea. Positive correlation between serum fractalkine CX3CL1 level and age, duration of disease, urea, creatinine, A/C ratio, HbA1C and IgG in patient with DN. Serum CX3CL1 level, urinary IgG were significantly increased with the progress of nephropathy so these integrated biomarkers could be used as good predictors for early identification of nephropathy. But miRNA- 152-3p has inadequate prognostic indicator for ESRD progression.

Study of Monocyte Subsets and Their Surface Expression of CD86 and Serum IL-17 Compared to Serum Procalcitonin as Markers of Early Neonatal Sepsis.

INTRODUCTION: Neonatal sepsis can quickly progress to multi-organ failure with high morbidity and mortality, making early diagnosis mandatory. Although being the gold standard, the long duration of blood culture may lead to hazardous neonatal complications. Sepsis activates monocytes and changes their subset distribution with the resultant activation of lymphocytes and adaptive immune cells changing the plasma cytokines levels. SUBJECTS AND METHOD: Percentages of monocytes subsets, pattern of monocytes surface CD86 expression and serum IL-17 compared to serum procalcitonin were measured in 30 neonates with early sepsis and compared with age and sex matched 30 apparently health neonates as a control group. RESULTS: Gestational age, neonatal weight and hemoglobin concentration were significantly low in septic neonates vs the control group. Percentages of intermediate, nonclassical and CD86 positive monocytes, the mean fluorescence intensity of CD16 on CD16 positive monocytes, and serum levels of CRP, IL-17 and procalcitonin were significantly increased in septic neonates compared with the control group. CONCLUSION: Early neonatal sepsis was associated with increasing the percentage of CD86 positive monocytes. Serum IL-17 levels were positively correlated with increased serum procalcitonin.

T-Natural Killers and Interferon Gamma/Interleukin 4 in Augmentation of Infection in Foot Ulcer in Type 2 Diabetes.

BACKGROUND: The link between immune system and type 2 diabetes mellitus (T2DM) pathogenesis attracted attention to demonstrate the role of immune cells and their secreted cytokines in T2DM development and its subsequent foot complications. OBJECTIVE: To investigate the relation between T Natural killer cell (TNK) %, Interleukin 4 (IL4) and Interferon gamma (IFN-γ) and diabetic foot infection (DFI) development in patients with diabetic foot ulcer (DFU). PATIENTS AND METHODS: Ninety patients with diabetes were included in this work, divided as T2DM group (n=30), DFU group (n=30), and DFI group (n=30). TNK% was detected using flow cytometry. Serum IL4 and IFN-γ were measured by ELISA. Diabetes biochemical parameters were also analyzed. RESULTS: Significant decrease was detected in TNK% and IFN-γ in DFI group compared to other 2 groups (P<0.001). Significant decrease was detected in serum levels of IL4 in DFI group compared to T2DM group (P=0.006). IFN-γ/IL4 was significantly decreased in DFI compared to DFU group (P=0.020). There was a significant correlation of TNK% with both IL4 and IFN-γ (r=0.385, P<0.001; r=0.534, P<0.001, respectively). Significant negative correlation of TNK% with HbA1c and LDL was revealed (r=-0.631, P<0.001; and r=-0.261, P=0.013, respectively), while a positive correlation was seen with HDL (r=0.287, P=0.006). A significant negative correlation of IL4 with HbA1c  was found (r=-0.514, P<0.001;. As for IFN-γ, a significant negative correlation with HbA1c and LDL was detected (r=-0.369, P< 0.001; r=-0.229, P=0.030). TNK % and IFN-γ level showed negative correlations with disease duration/year (r=-0.546, P< 0.001; r=-0.338, P=0.001,respectively). CONCLUSION: Decline in TNK frequency has essential role in T2DM pathogenesis and subsequent foot complications. Downregulation of TNK% and IFN-γ level have potential roles in predicting infection of diabetic ulcer and are correlated with disease duration.

Peripheral Polyneuropathy and Cognitive Impairment in Type II Diabetes Mellitus.

BACKGROUND: Neuropathy is one of most common complications in diabetic patients. Diagnosis of diabetic neuropathy is essential for decreasing the rate of the disability and death. Neuron-specific enolase (NSE) is released from damaged neuronal cells and enters the blood circulation through an injured blood brain barrier. Therefore, serum NSE can reflect the damage of neurons and brain tissue. OBJECTIVE: To evaluate peripheral polyneuropathy and cognitive function in Type 2 Diabetes Mellitus (T2DM) and correlate them with NSE level as a possible biomarker of diabetic neuropathy. SUBJECTS AND METHODS: Forty five T2DM patients with polyneuropathy were randomly recruited in this study compared to 45 healthy age and sex matched subjects as a control. Patients group were divided into two subgroups, 24 diabetic patients with painful peripheral neuropathy and 21 with painless peripheral neuropathy. All were subjected to clinical assessment by diabetic neuropathy symptom score, Dyck neuropathy grading, Mini-Mental State Examination (MMSE), assessment of HbA1c, NSE biomarker and neurophysiological assessment (nerve conduction study (NCS), event related potential (P300wave) and somatosensory evoked potential (SSEP) of the right median nerve). RESULTS: There were significant decrease in cognitive functions in diabetic patients compared to controls and a significant increase in NSE in diabetic patients. There were no significant difference between patients with painless and painful diabetic neuropathy as regard MMSE, HbA1c and NSE. There were significant correlation of P300 in diabetic patients with HbA1c and NSE. CONCLUSION: Neurophysiological assessment of diabetic patients by NCS, SSEP and P300 have well evaluation of cognitive functions, painless, and painful diabetic polyneuropathy. NSE is a beneficial biomarker in diabetic patients to pick up neurological complications.

Apelin and chemerin as promising adipokines in children with type 1 diabetes mellitus.

BACKGROUND: Type 1 diabetes mellitus (T1DM) is one of the most common chronic diseases in children that may be due to micro or macrovascular complications. Diabetic renal disease or nephropathy is a common complication of DM, clinically silent and the only detectable abnormality due to the presence of microalbuminuria. SUBJECTS AND METHODS: This study was a case-control study. Participants were classified into two groups. The first group included 40 children with T1DM and the second group included 30 matched healthy controls. Serum apelin (APLN), chemerin, cholesterol, and triglycerides (TG) levels were measured for each case. Also, albumin/creatinine ratio was analyzed in random urine sample. RESULTS: Comparison between T1DM patients and controls revealed that serum apelin, chemerin, cholesterol, TG levels, and albuminuria were significantly increased in cases compared to their controls. Significant positive correlations were found between HbA1c% and albuminuria for APLN and chemerin in the diabetic group. Whereas significant negative correlations were found between apelin and glomerular filtration rate (GFR). CONCLUSION: Increased levels of serum apelin and chemerin in T1DM patients may be considered as promising adipokines for the development of diabetic complication.