Gene Polymorphism of Interleukin-4, Interleukin-4 Receptor and STAT6 in Children with Atopic Dermatitis in Taif, Saudi Arabia.

AIM OF STUDY: This work was performed to evaluate the level of IL-4, and to clarify the role of IL-4 gene polymorphism at position cytosine -590-to-thyamine (C-590T), IL-4Rα gene polymorphism at position adenine +4679-to-guanine (A+4679G) [isoleucine-50-valine (I50V)] and STAT6 gene polymorphism at position guanine 2964-to-adenine (G2964A) in Saudi children with non-atopic dermatitis (non-AD) and atopic dermatitis (AD) to identify their role in the pathogenesis of these diseases. SUBJECTS AND METHODS: This study included 150 children: 50 healthy children as controls, 50 with non-AD, and 50 with AD. They were subjected to full clinical examination, complete blood picture, skin prick test, and determination of serum interleukin-4 (IL-4) and total immunoglobulin-E (IgE) levels. Detection of interleukin-4 gene (C-590T), interleukin-4 receptor alpha gene (A+4679G) (I50V), and STAT6 gene (G2964A) polymorphisms were performed by PCR-based restriction fragment length polymorphism (PCR-RFLP). RESULTS: There was a significant (P < 0.01) association between genotype and allele frequencies of IL-4Rα (A+4679G) (I50V) polymorphism in the AD group (but not non-AD group). Moreover, there was a significant association between genotype and allele frequencies of the STAT6 (G2946A) polymorphism in the non-AD (P < 0.05) and AD (P < 0.01) groups. On the other hand, there was no significant association between genotype and allele frequencies of the (C-590T) polymorphism in the non-AD group and AD group. There was a significant (P < 0.001) higher total IgE level in patients compared to the controls. Moreover, the mean values of total IgE were significantly different among the different allelic variants of (C-590T), (I50V), (G2964A) polymorphisms of IL-4, IL-4Rα, and STAT6 genes, respectively, in all the studied groups. On the other hand, there was no significant difference of serum IL-4 levels among all the studied patients, or among the different allelic variants of (C-590T), (I50V), (G2964A) polymorphisms of IL-4, IL-4Rα, and STAT6 genes, respectively. CONCLUSION: IL-4Rα gene (I50V) and STAT6 gene (G2964) polymorphisms may play a role in development of eczema; however, the IL-4 gene polymorphism (C-590T) had no relationship with susceptibility to the disease among Saudi children.

Hepatoprotective activity of quercetin against acrylonitrile-induced hepatotoxicity in rats.

Acrylonitrile is a potent hepatotoxic, mutagen, and carcinogen. A role for free radical-mediated lipid peroxidation in the toxicity of acrylonitrile has been suggested. The present study was designed to assess the hepatoprotective effect of quercetin against acrylonitrile-induced hepatotoxicity in rats. Liver damage was induced by oral administration of acrylonitrile (50 mg/kg/day/5 weeks). Acrylonitrile produced a significant elevation of malondialdehyde (138.9%) with a marked decrease in reduced glutathione (72.4%), and enzymatic antioxidants; superoxide dismutase (81%), and glutathione peroxidase (53.2%) in the liver. Serum aspartate aminotransferase, alanine aminotransferases, direct bilirubin, and total bilirubin showed a significant increase in acrylonitrile alone treated rats (115.5%, 110.8%, 1006.8%, and 1000.8%, respectively). Pretreatment with quercetin (70 mg/kg/day/6 weeks) and its coadministration with acrylonitrile prevented acrylonitrile-induced alterations in hepatic lipid peroxides and enzymatic antioxidants as well as serum aminotransferases and bilirubin. Histopathological findings supported the biochemical results. We suggest that querectin possess hepatoprotective effect against acrylonitrile-induced hepatotoxicity through its antioxidant activity.

Bone mineral density and vitamin D receptor polymorphism in beta-thalassemia major.

Osteoporosis is the most prevalent bone complication in beta-thalassemic patients despite regular transfusions and iron chelation therapy. Although its etiology is multi-factorial, genetic factors play an important role in pathogenesis. These factors have not yet been clearly defined, however, osteoporosis may be related to vitamin D receptor gene BsmI polymorphism. In this study, BsmI vitamin D receptor gene polymorphism was analyzed using polymerase chain reaction and BsmI restriction fragment length polymorphism in 42 regularly treated-beta-thalassemic patients of different ages. Bone mineral density was measured by peripheral quantitative ultrasound at the heel of the foot. Serum levels of alkaline phosphatase, calcium, phosphorus, ferritin and 25-hydroxyvitamin D3 were determined. Patients were divided into two groups according to pubertal signs: group I (22 children), and group II (20 adolescents and adults). The Z-scores of bone mineral density in both groups were -1.32 +/- -0.9 and -2.30 +/- -1.02 respectively, with a significant difference between the two groups. The height standard deviation and 25-hydroxyvitamin D3 were significantly decreased in group II compared to group I. Moreover, significantly lower bone mineral density and height standard deviation were detected among patients with BB vitamin D receptor genotype. Therefore, this genotype may be considered as a risk factor for osteoporosis in beta-thalassemic patients.

Experimental diabetic nephropathy can be prevented by propolis: Effect on metabolic disturbances and renal oxidative parameters.

Oxidative stress may play a key role in the pathogenesis of diabetic nephropathy. Propolis and its extract have antioxidant properties. The effect of ethanolic extract of propolis against experimental diabetes mellitus-associated changes was examined. Diabetes was induced experimentally in rats by i.p. injection of streptozotocin (STZ) in a dose of 60 mg/kg bwt for 3 successive days. Blood urea nitrogen (BNU), creatinine, glucose, lipid profile, malondialdehyde (MDA) and urinary albumin were measured. Superoxide dimutase (SOD), glutathione (GSH), catalase (CAT) and MDA were measured in the renal tissue. The results showed decreased body weight and increased kidney weight in diabetic animals. Compared to the control normal rats, diabetic rats had higher blood glucose, BNU, creatinine, total cholesterol, triglycerides, low-density lipoprotein-cholesterol (LDL-C), MDA and urinary albumin and lower high-density lipoprotein-cholesterol (HDL-C) levels. Moreover, renal tissue MDA was markedly increased while SOD, GSH and CAT were significantly decreased. Oral administration of propolis extract in doses of 100,200 & 300 mg/kg bwt improved the body and kidney weights, serum glucose, lipid profile, MDA and renal function tests. Renal GSH, SOD and CAT were significantly increased while MDA was markedly reduced. These results may suggest a strong antioxidant effect of propolis which can ameliorate oxidative stress and delay the occurrence of diabetic nephropathy in diabetes mellitus.

Prevalence of mupirocin and chlorhexidine resistance among methicillin-resistant coagulase-negative staphylococci isolated during methicillin-resistant Staphylococcus aureus decolonization strategies.

BACKGROUND: The widespread of methicillin-resistant Staphylococcus aureus (MRSA) antimicrobial decolonization in the clinical setting may lead to an increase in the prevalence of multiresistance to coagulase-negative staphylococci (CoNS) owing to their selection. This study aimed to investigate the impact of MRSA decolonization strategies, using mupirocin and chlorhexidine, on their CoNS susceptibility. METHODS: A total of 312 CoNS isolates were collected before starting the decolonization protocols "baseline strains" (BLS) group, 330 isolates were collected after application of the targeted decolonization protocol "targeted decolonization strains" group, and 355 isolates were collected after application of the universal decolonization protocol "universal decolonization strains" group. Methicillin-resistant CoNS (MR-CoNS) were identified and tested for mupirocin and chlorhexidine susceptibilities. Heptaplex polymerase chain reaction assay was applied for simultaneous screening for chlorhexidine (CHX-R) and mupirocin resistance (Mu-R) genes. RESULTS: Mu-R prevalence of MR-CoNS among the BLS group was considered moderate (9.1%); however, CHX-R in the BLS group was 5.8%, the rate of which significantly increased among the universal decolonization strains group. DISCUSSION: Both MRSA decolonization strategies have an additional benefit in reducing the prevalence of MR-CoNS. The prevalence Mu-R rate didn't change significantly during either of the MRSA decolonization practices that may be due to the local nature of mupirocin application on the nasal mucosa only. In contrast CHX-R that was found to be significantly higher among the UDS group. CONCLUSIONS: Our findings indicate that both MRSA decolonization strategies have an additional benefit in reducing the prevalence of MR-CoNS. Although the universal MRSA decolonization has superior efficacy in decolonization of CoNS, it may increase the risk of selecting CHX-R and Mu-R. In addition, other potential resistance genes should be studied.

Risk factors and molecular features of extended-spectrum beta-lactamase producing bacteria at southwest of Saudi Arabia.

OBJECTIVES: To explore the risk factors, the prevalence rate, and gene types of extended-spectrum beta-lactamase (ESBL)-producing bacteria as the causative agents of infection at King Abdulaziz Specialist Hospital (KAASH), Taif, Kingdom of Saudi Arabia. Methods: This was a retrospective study conducted during the period between February 2017 and January 2018. All samples obtained from the KAASH were analyzed. The MicroScan Walkaway System, bacteriological examination and double disk synergy tests were used to detect ESBL-producing bacteria. To identify ESBL genes, the polymerase chain reaction (PCR) technique was used. Results: The ESBL phenotype was detected in 351 of 1151 isolates (30.5%); Escherichia coli (E. coli) (62.7%) and Klebsiella pneumoniae (K. pneumoniae) (23.6%) were the most prevalent. The highest proportion of ESBL specimens was found in urine (62%.5), and these organisms were mainly isolated from the female medical ward (20.2%). Based on the statistical analysis, lung diseases, renal diseases, diabetes and heart diseases contributed to the spread of ESBL infections. Amikacin, imipenem, meropenem and tigecycline were found to be effective in overcoming ESBL infections; however, these antibiotics may be inappropriate for new strains of K.pneumoniae. The distribution of the blaCTX-M gene was high (87%), compared with blaTEM (74.9%) and blaSHV (29.4%).   Conclusion: These data provide new epidemiological information about the prevalence of ESBL-producing organisms among patients in KAASH, Taif, Saudi Arabia. In addition, this study identified the clonal nature of isolated E.coli and K.pneumoniae.

T-Regulatory Cell Subsets in Children with Type 1 Diabetes Mellitus: Relation to Control of the Disease.

BACKGROUND: Type 1 diabetes mellitus is described as a chronic metabolic disorder characterized by aggressive immune &#946;-cell destruction. There are a number of varied immune mechanisms for sustaining self-tolerance in opposition to the autoimmune disorders. A recessive tolerance is accomplished by thymic gland via a negative assortment of different clones, while a dominant tolerance is accomplished by the regulatory T cells (Treg) in the periphery. Treg (CD4+ CD25+FOXP3+) are subsets of T cells which have an essential role in maintaining tolerance. OBJECTIVE: To evaluate peripheral Treg (CD4+; CD25+; FOXP3+) in children cohort with T1DM. METHODS: This study included 64 children diagnosed with T1DM and 35 age- and sex-matched healthy children as controls. All children were clinically evaluated and subjected to assessment of complete blood count (CBC), glycated hemoglobin, surface and cytoplasmic detection of Treg by flow cytometry. RESULTS: This study showed that the frequency of Treg (CD4+; CD25+; FOXP3+) was significantly lower in diabetic children than with normal controls (P<0.001). There was a significant (P <0.001) reduction in the Treg (CD4+; CD25+; FOXP3+) in T1DM children with uncontrolled (Hemoglobin A1c>7%) as compared to those with controlled (Hemoglobin A1c<7%) disease. CONCLUSION: Diminished Treg in T1DM proved that auto-reactivation of T-cell as a result of the breakdown of immune tolerance takes part in the elaboration of autoimmune disorders as T1DM. Treg may be used in immunotherapy, thus preventing T1DM development due to its pivotal role in immune tolerance.

Phenotypic and molecular characterization of HA-MRSA in Taif hospitals, Saudi Arabia.

INTRODUCTION: Methicillin-resistant S. aureus (MRSA) is one of the most important organisms causing hospital-acquired infections worldwide. Molecular analysis of MRSA strains from Taif, Saudi Arabia, had not been previously done. Phenotypic and molecular characteristics of MRSA isolated from Taif hospitals were investigated. METHODOLOGY: This study involved S. aureus strains isolated from different clinical samples from Taif hospitals. MRSA strains were identified and antimicrobial susceptibility profiles were determined. Multiplex polymerase chain reaction (PCR) was used to identify S. aureus-specific sequence, mecA genes, and type of staphylococcal cassette chromosome mec (SCCmec). MRSA strains were typed using coagulase gene polymorphism. RESULTS: In total, 390 strains of S. aureus were isolated, and 58 MRSA strains - 40 hospital-acquired MRSA (HA-MRSA) and 18 community-acquired MRSA (CA-MRSA) - were detected. HA-MRSA strains included three SCCmec types, while CA-MRSA strains included two SCCmec types. PCR amplification and restriction of the coagulase gene of the 58 MRSA isolates showed nine different patterns, while three strains were non-typable. HA-MRSA strains showed seven distinct restriction fragment length polymorphism (RFLP) patterns; the most frequent was pattern 2 (15 isolates), followed by patterns 1 and 4 (5 isolates each). CA-MRSA showed five RFLP patterns; the most frequent was pattern 3 (7 isolates) followed by pattern 8 (6 isolates). CONCLUSIONS: HA-MRSA strains were more common than CA-MRSA strains. SCCmec typing and coagulase gene polymorphism analysis may be useful methods for studying clonal relatedness of isolates and for discriminating between HA-MRSA and CA-MRSA.

Evaluation of recent methods versus conventional methods for diagnosis of early-onset neonatal sepsis.

INTRODUCTION: Hospital-acquired infections continue to be a major public health problem, especially among neonates. Large proportions of infants are admitted to neonatal intensive care units (NICUs) and receive potent systemic antibiotics while the diagnostic work-up is still in progress. This study aimed to evaluate the recent methods for diagnosing neonatal sepsis (NS) and compare them to conventional diagnostic work-up. METHODOLOGY: The study included 100 neonates divided into three groups: proven early-onset NS, clinical early-onset NS, and negative infectious status. Bacterial DNA was detected in the blood by broad-range 16S rDNA polymerase chain reaction (PCR). Markers for diagnosis of bacterial infection, which includedprocalcitonin (PCT), interleukin-6 (IL-6), and highly sensitive C-reactive protein (hs-CRP), were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Blood culture was positive in 25 cases, while PCR for 16S rDNA was positive in 32 cases. Hs-CRP was significantly elevated in 30 patients in group 1, 35 patients in group 2, and 8 patients in group 3. IL-6 was significantly elevated in 28 patients in group 1, 24 patients in group 2, and 9 patients in group 3. PCT was found to be significantly elevated in 29 patients in group 1, 31 patients in group 2, and 2 patients in group 3. CONCLUSIONS: The16S rDNA PCR assay was more sensitive than blood culture. The combination of markers (hs-CRP, PCT, and IL-6) is better than single markers to diagnose sepsis. PCT had greater diagnostic value than did hs-CRP and IL-6, while IL-6 was better for diagnosis of neonatal infection.

Association of Helicobacter pylori cagA Gene with Gastric Cancer and Peptic Ulcer in Saudi Patients.

This study was conducted to assess the relationship between occurrence of gastric cancer and peptic ulcer, and the presence of H. pylori cagA gene and anti-CagA IgG, and to estimate the value of these antibodies in detecting infection by cagA gene-positive H. pylori strains in Saudi patients. The study included 180 patients who were subjected to upper gastrointestinal endoscopy in Taif province and Western region of Saudi Arabia (60 gastric cancer, 60 peptic ulcer, and 60 with non-ulcer dyspepsia). Gastric biopsy specimens were obtained and tested for H. pylori infection by rapid urease test and culture. PCR was performed on the isolated strains and biopsy specimens for detection of the cagA gene. Blood samples were collected and tested for CagA IgG by ELISA. H. pylori infection was detected among 72.8% of patients. The cagA gene and anti-CagA IgG were found in 63.4% and 61.8% of H. pylori-infected patients, respectively. They were significantly (p < 0.01) higher in patients with gastric cancer and peptic ulcer compared with those with non-ulcer dyspepsia. Detection of the CagA IgG was 91.6% sensitive, 89.6% specific, and 90.8% accurate compared with detection of the cagA gene. Its positive and negative predictive values were 93.8% and 86%, respectively. The study showed a significant association between the presence of the cagA gene and gastric cancer and peptic ulcer disease, and between anti-CagA IgG and the cagA gene in Saudi patients. However, a further larger study is required to confirm this finding.

A study of the role of IL-12 in pulmonary tuberculosis using the whole blood flowcytometry technique.

Pulmonary tuberculosis remains a major health problem. It is caused by Mycobacterium tuberculosis, which elicits a T-cell dependent immune response, initiated by monocytes through a large number of cytokines of which interleukin-12 is thought to play a critical role in initiation and regulation of T-helper (Th-1) like responses. To better understand the role of IL-12 in pulmonary tuberculosis patients, intracellular IL-12 in peripheral blood-derived monocytes was examined by flowcytometery. The percentage of monocytes producing IL-12 was measured after invitro stimulation of heparinized whole blood with mycobacterial protein antigens (culture filtrate). Of the 22 active tuberculosis patients, 17 were recent cases and 5 recurrent cases. Healthy controls were 14 individuals with detectable reaction to purified protein derivative (PPD+) and 14 without detectable reaction to PPD. The role of different factors affecting disease outcome such as treatment, age, gender, smoking, severity of disease and presence of other complications on the percentage of monocytes producing IL-12 was studied. Recurrent TB patients had a higher number of monocytes producing IL-12 in unstimulated cultures compared to other groups (P < 0.001). However, after in vitro stimulation there was a significant decrease in the number of monocytes producing IL-12 in recurrent TB patients as compared to recently diagnosed TB patients and healthy PPD+ individuals (P < 0.001). Antituberculosis chemotherapy was the only factor that had significant effect on the percentage of monocytes producing IL-12 (p < 0.05) while other studied factors did not show significant effect (p > 0.05). It is concluded that IL-12 plays a prominent regulatory role in tuberculosis.

Human natural killer T cells (NKT), NK and T cells in pulmonary tuberculosis: potential indicators for disease activity and prognosis.

One third of the world's population is infected with Mycobacterium tuberculosis (MTB). However, active disease can develop only in a small percentage, when the immunity is weakened. The acquired immune response to MTB is primarily mediated by T cells. Natural killer (NK) cells play a central role in innate immunity to microbial pathogens. Human NKT cells have characteristics of both T and NK cells and also exhibit antimycobacterial activity. This work aimed to enumerate T, NK and NKT cells in active pulmonary TB compared with healthy controls and to study the correlation between these cells with different factors affecting prognosis of pulmonary TB as disease severity, complications or associated diseases, antitubrculosis chemotherapy, and age & gender. Of the 22 active tuberculosis patients examined, 17 were recent cases and 5 recurrent. Healthy controls were divided into 14 individuals with detectable reaction to purified protein derivative (PPD+) and 14 individuals without detectable reaction to PPD-. The percentages of T, NK and NKT cells in erythrocyte-lysed whole blood samples were determined using flowcytometry. The percentage of NKT cells was significantly higher among the recently diagnosed MTB cases as compared with both PPD+ (P < 0.01) and PPD- (P < 0.01) healthy controls, while no significant difference could be found in the percentages of T or NK cells among these groups. However, comparing recurrent cases with recently diagnosed cases showed a significant difference only in the percentage of T cells (P < 0.01). There was also a significant difference in the percentage of T cells according to severity of disease (P < 0.01) and in the association of diabetes mellitus (P < 0.01). Age, gender and treatment with antituberculosis chemotherapy had no effect on the percentages of T, NK or NKT cells. It is concluded that T and NKT cells play an important role in immunity against TB. In active pulmonary tuberculosis, increased T cell count points to severity of the disease, while their reduced count predicts bad prognosis. Human NKT cell count is a marker of disease activity. Enumeration of these cells in peripheral blood can be used as a non-invasive prognostic indicator for patients with active pulmonary TB.