The Diagnostic Utility of Natural Killer Cell Subsets in Deep Vein Thrombosis.

Background: Natural killer (NK) cells are important components of adaptive and innate immune responses. NK cell subsets have different functions and may play a role in vascular disorders. This study aimed to evaluate the proportions of NK cells and their subsets to determine whether they can be used as markers of venous thrombosis and to identify whether there was a link between NK cell proportion and citrullinated histone (H3) levels. Patients and Methods: This study included 100 participants divided into Group I (n=50, patients with deep venous thrombosis (DVT)) and Group II (n=50, age- and sex-matched healthy controls). Group I was further categorized into Group Ia (n=25, patients with acute DVT) and Group Ib (n=25, patients with chronic DVT). The proportions of NK cells and their subsets were evaluated by flow cytometry using CD3/CD16/CD56. The levels of citrullinated histones (H3) were estimated using enzyme-linked immunosorbent assay (ELISA). Results: Compared to the control group, DVT patients had a significantly lower proportion of (CD56 dim/CD16+) NK cells, a significantly higher proportion of (CD56-/CD16+) NK cells and a high level of citrullinated histone (H3). Conclusion: NK cell subsets and citrullinated histone (H3) could be used as markers for DVT and as targets for therapeutic drugs to inhibit the formation or progression of thrombosis.

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.

Ultrastructural changes and IgA modulatory effect of commercial prebiotic and probiotic in murine giardiasis.

Giardiasis, a parasitic infection of the gastrointestinal tract, is prevalent worldwide. The integrity of the intestinal epithelial barrier plays an important defensive role in giardiasis, and as Oral supplementation with prebiotics and probiotics is known to reinforce the intestinal barrier in many gastrointestinal diseases, this study assessed the effects of prebiotic and probiotic supplementation in giardiasis and compared the results with those obtained after nitazoxanide therapy. Swiss albino male lab-bred mice (n = 50) were divided into three major groups; Group I (control group), i.e., negative (noninfected nontreated) and positive controls (infected nontreated); Group II (preventive group), in which mice were provided prebiotic, probiotic, or a combination for 7 days before of infection, and Group III (therapy group), in which mice were administered prebiotic, probiotic, combined supplements and nitazoxanide from day 12 post-infection. The assessment was achieved through Giardia cyst count, histopathological examination and ultrastructure study. Also, Serological and immunohistochemical parameters were done to evaluate the modulation of IgA levels. Oral supplementation with prebiotic and probiotic, either before or after infection (in preventive or therapy groups respectively) resulted in a significant reduction in Giardia cyst shedding. Remarkable histological and ultrastructure improvement in the intestinal changes, along with a significant increase in the serological and immunohistochemical IgA levels, were seen in mice provided combined supplements and nitazoxanide (in therapy group). Thus, our results indicate that combined prebiotic and probiotic supplementation has promising anti-Giardia activity and that it can restore intestinal structures and modulate IgA response, apart from providing synergistic effects when added to nitazoxanide.

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.

MicroRNA-146a-5p and microRNA-210-3p Correlate with T Regulatory Cells Frequency and Predict Asthma Severity in Egyptian Pediatric Population.

Background: Severe bronchial asthma (BA) affects 5-10% of children, which imposes socioeconomic burden. Therefore, it is crucial to identify biomarkers for risk stratification in children with BA. T regulatory cells (Tregs) play a balancing role in allergic response regulation. We aimed to investigate the relationship between Treg, miR-210-3p, and miR-146a-5p in relation to asthma phenotypes in search of novel biomarkers of disease severity. Methods: This study included 50 children with BA classified into Group 1 (n = 25) children with mild to moderate asthma and Group 2 (n = 25) children with severe asthma. In addition to 26 control subjects. Flow cytometry was used to detect Tregs. Plasma miR-210-3p and miR-146a levels were determined using quantitative real-time PCR. Patients' FEV1 (Forced Expiratory Volume in the first second) was measured. Results: miR-210-3p level correlated negatively with Treg frequency (r = -0.828, P < 0.001) and FEV1 (r = -0.621, P < 0.001). The level of miR-146a-5p positively correlated positively with Treg% (r = 0.303, P = 0.032). ROC curve analysis revealed that miR-210-3p was the most sensitive biomarker of severity, with the area under curve (AUC) = 0.923, 96% sensitivity, and 60% specificity. According to multivariate analysis, miR-210-3p is an independent risk factor for BA severity [OR =3.119, P = 0.030], while miR-146a-5p is a protective factor [OR =0.811, P = 0.049]. Conclusion: Treg frequency is linked to FEV1, miR-146a-5p and miR-210-3p in childhood BA. Upregulation of miR-210-3p is a sensitive biomarker and an independent risk factor for BA severity in Egyptian children.

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.

Apoptotic changes in the intestinal epithelium of Cryptosporidium-infected mice after silver nanoparticles treatment versus nitazoxanide.

Cryptosporidium has been identified as one of the prevalent opportunistic parasites that cause diarrhea, which may be persistent and fatal. Current chemotherapeutic agents, including nitazoxanide (NTZ), are frequently associated with therapeutic failure, and their roles in the induction of apoptosis in cryptosporidiosis remain to be a topic of debate. Thus, this study aimed to assess the apoptotic changes in cryptosporidiosis in immunocompetent (IC) and immunosuppressed (IS) mice after treatment with silver nanoparticles (AgNPs) and NTZ either alone or after loading. In total, 120 laboratory-bred Swiss albino mice were divided into two groups. Group A included IC mice, while Group B included IS mice. Both groups were divided into six subgroups: noninfected nontreated, infected nontreated, infected AgNP-treated, infected NTZ-treated, infected AgNP-loaded NTZ (full-dose)-treated, and infected AgNP-loaded NTZ (half-dose)-treated. The assessment was achieved through parasitological, histopathological, and apoptotic marker expression evaluation. AgNP-loaded NTZ (different doses) treatment showed the highest oocyst shedding reduction and remarkable improvement in histopathological changes, followed by individual treatment with NTZ and then AgNPs in IC and IS mice. Results of apoptotic marker expression revealed that AgNP-loaded NTZ treatment exhibited a promising role in regulating apoptotic changes in cryptosporidiosis through the expression of the lowest levels of cytochrome C and caspase-3 in IC and IS mice at the end of the experiment. Therefore, AgNP-loaded NTZ can be a potential therapeutic agent against cryptosporidiosis for IC and IS mice.

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.

Vegetable-transmitted parasites among inhabitants of El-Prince, Alexandria and its relation to housewives' knowledge and practices.

In the present study, stool samples were obtained from 575 individuals from El-Prince, a suburban village of Alexandria, Egypt to detect helminthic eggs and protozoan cysts. The results showed that, 35.7% of the sample had at least one vegetable-transmitted parasite, the highest percentage (46.4%) was in the age group of 6 to less than 15 years. The nematode Ascaris lumbricoides was the most prevalent in 18.4%, followed by the protozoan Giardia lamblia in 10.4%. Fasciola eggs were found in 2.4% of samples. Knowledge and practices of housewives concerning Fasciola and its source of transmission and methods of washing leafy vegetables was obtained through house-to-house interviews with 303 housewives. Knowledge that leafy vegetables were a source of Fasciola infection was indirectly proportional with better in younger housewives and those of secondary or higher education. More than half of the interviewed housewives (57.7%) washed leafy vegetables under running tap water and 32.7% soaked them in tap water. Only 9.6% soaked them in water mixed with a substance as vinegar, lemon juice or common salt. Only 5% of those who were infected with vegetable transmitted parasites washed vegetables by soaking in water with an added substance compared to 19.6% of parasite free housewives. Most of those adding a substance to soaking water (89.7%) used vinegar. Results of the study revealed that a serious and consistent effort through public health activities is essential to educate housewives about vegetable-transmitted parasites, their transmission and ways of prevention.

Current status of S. mansoni infection in El-Prince village, Alexandria Governorate.

The present work was implemented to determine the current status of Schistosoma mansoni infection in "El-Prince" village, near Alexandria, which was studied before as a control village between 1985 and 1990. Stool examination was performed on 571 of the inhabitants by Kato thick smear technique and intensity of infection was determined. Prevalence was found to be 15.4% with a percentage decrease of 53% from the 1990 survey. The geometric mean egg count (GMEC) ranged from 19.05 eggs/gram of stools in children aged five years or less to 81.86 in the oldest group greater than 50 years of age and averaged 42.26. Age stratified prevalence of infection peaked at 31.4% in the (20 to 25)-year-old age group. High prevalence of 23.6% was also observed in the age group from 36 to 50 years. Infection in males was higher than females (17.8% vs.13.5%). However the difference was not statistically significant (chi2 = 2.2, P = 0.0086). Risk factors significantly associated with the infection were an age more than 5 years, male gender, males with water contact activities and having done a previous laboratory analysis for schistosomiasis. The community category of the village changed from high prevalence in 1990 to moderate prevalence in 2002. Most of the infected individuals (85%) had light intensity of infection (less than 99 eggs /g stools). Active screening and treatment programs should be maintained in the area. More control measures should be directed towards older people who should be included in evaluation of control programs.