The potential role of serum expression profile of long non coding RNAs, Cox2 and HOTAIR as novel diagnostic biomarkers in systemic lupus erythematosus.

BACKGROUND: The role of the long non-coding RNAs (lncRNAs) in the pathogenesis of systemic lupus erythematosus (SLE) is mostly unknown, despite increasing evidence that lncRNAs extensively participate in physiological and pathological conditions. AIM: To detect the level of lncRNA-Cox2, HOTAIR, IL-6, and MMP-9 in the serum of SLE patients and to correlate these levels with disease activity and patients' clinical and laboratory data to evaluate the value of these biomarkers for SLE diagnosis and assessment of disease activity. METHODS: Blood samples from 58 SLE patients, and 60 healthy controls (HCs) were used for detection of lncRNAs-Cox2 and HOTAIR expression levels by real-time polymerase chain reaction. Both IL-6 and MMP-9 serum levels were assayed by enzyme-linked immunosorbent assay. Lupus activity was assessed with the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). RESULTS: The serum expression levels of lncRNA-Cox2 and HOTAIR were significantly up-regulated in SLE patients vs HCs (fold change [median (IQR) was 1.29(0.81-1.71, P<0.0001) and 2.68(0.95-3.67), P = 0.038) for lncRNA-Cox2 and HOTAIR, respectively. Serum levels of both IL-6 and MMP-9 were significantly high in SLE patients compared with HCs (P≤0.001 for each). The up-regulated lncRNA-Cox2 was positively associated with the presence of neurological manifestations in SLE patients (P = 0.007). Furthermore, HOTAIR expression level had significantly positive correlation with IL-6 (r = 0.578, P<0.0001), MMP-9 level (r = 0.762, P<0.0001), nephritis grades (r = 0.296, P = 0.024) and proteinuria (r = 0.287, P = 0.035). LncRNA-Cox2 showed sensitivity and specificity 72.4%, and 100.0% respectively. HOTAIR sensitivity was 60.3%, and specificity was 100.0%. By multiple logistic regression analysis, lncRNA-Cox2 and HOTAIR were found as SLE independent predictors. CONCLUSION: LncRNA-COX2 and HOTAIR can be used as new non-invasive biomarkers for the diagnosis of SLE.

Diagnostic value of monocyte chemoattractant Protein-1, soluble mannose receptor, Presepsin, and Procalcitonin in critically ill children admitted with suspected sepsis.

INTRODUCTION: The differentiation between systemic inflammatory response syndrome and sepsis is very important as it determines essential treatment decisions, such as selection, initiation, and duration of antibiotic therapy. OBJECTIVES: We aimed to investigate the diagnostic value of Procalcitonin, Monocyte Chemoattractant Protein-1, soluble Mannose Receptor, Presepsin as early biomarkers of pediatric sepsis in comparison to systemic inflammatory response syndrome in severely ill children. PATIENTS AND METHODS: This study included 58 children diagnosed as sepsis (group 1), 24 children with systemic inflammatory response syndrome without infection (group 2), and 50 healthy children as controls (group 3). All the plasma levels of the studied biomarkers were measured and ROC curves were created for all the tested parameters to discriminate between sepsis and SIRS. RESULTS: The area under the curve for Monocyte Chemoattractant Protein-1 was 0.926 (0.846-0.927) with sensitivity 100% and specificity 62.5%. The soluble Mannose Receptor had the highest sensitivity (100%), with AUC equals 1(.0.956-1.0) and specificity of 100%. The cut-off values for Procalcitonin, Presepsin, soluble Mannose Receptor, and Monocyte Chemoattractant Protein-1 and were: 0.62 ng/ml, 100 pg/ml, 13 ng/ml and 90 pg/ml, respectively. In septic cases, both soluble Mannose Receptor and Procalcitonin have positive correlations with the severity of sepsis, low Glasgow Coma Scale, ventilatory support, use of inotropic drugs and mortality rate (r = 0.950, 0.812, 0.795, 0.732 and 0.861respectively) for soluble Mannose Receptor and (0.536, 0.473, 0.422, 0.305 and 0.474 respectively) for Procalcitonin. CONCLUSION: Soluble Mannose Receptor, Presepsin, and Monocyte Chemoattractant Protein-1 can be used to differentiate between sepsis and SIRS in critically ill children.

miR-155 T/A (rs767649) and miR-146a A/G (rs57095329) single nucleotide polymorphisms as risk factors for chronic hepatitis B virus infection among Egyptian patients.

Genetic variants in microRNAs (miRNAs) can alter the miRNAs expression and/or function, accordingly, affecting the related biological pathways and disease risk. Dysregulation of miR-155 and miR-146a expression levels has been well-described in viral hepatitis B (HBV). In the current study, we aimed to assess rs767649 T/A and rs57095329 A/G polymorphisms in miR-155, and miR-146a genes, respectively, as risk factors for Chronic HBV (CHBV) in the Egyptian population. Also, we aimed to do in silico analysis to investigate the molecules that primarily target these miRNAs. One hundred patients diagnosed as CHBV and one hundred age and sex-matched controls with evidence of past HBV infection were genotyped for miR-155 (rs767649) and miR-146a (rs57095329) using real-time polymerase chain reaction. The rs767649 AT and AA genotypes in CHBV patients confer four folds and ten folds risk respectively, as compared to control subjects [(AOR = 4.245 (95%CI 2.009-8.970), p<0.0001) and AOR = 10.583 (95%CI 4.012-27.919), p<0.0001, respectively)]. The rs767649 A allele was associated with an increased risk of developing CHBV (AOR = 2.777 (95%CI 1.847-4.175), p<0.0001). There was a significant difference in the frequency of rs57095329 AG and GG genotypes in CHBV patients compared to controls. AG and GG genotypes showed an increase in the risk of developing CHBV by about three and six folds respectively [AOR = 2.610 (95%CI 1.362-5.000), p = 0.004] and [AOR = 5.604 (95%CI 2.157-14.563), p<0.0001].We concluded that rs57095329 and rs767649 SNPs can act as potential risk factors for the development of CHBV in the Egyptian population.

Tumor necrosis factor (TNF)-α- 308 G/A gene polymorphism (rs1800629) in Egyptian patients with alopecia areata and vitiligo, a laboratory and in silico analysis.

PURPOSE & METHODS: Several single-nucleotide polymorphisms (SNPs) in the promoter region of the TNF-α gene can cause variations in the gene regulatory sites and act as risk factors for some autoimmune disorders as alopecia areata (AA) and vitiligo. This study aimed to detect the serum TNF-α (sTNF) level (by ELISA) and the rs1800629 (by real-time PCR) among AA and vitiligo Egyptian patients and to determine their relation with disease duration and severity. In silico analysis of this SNP to study the molecular regulation of the mutant genotypes was also done. RESULTS: In AA patients, no risk was associated with the mutant genotypes vs. the normal genotype, or with A allele vs. G allele. The risk of vitiligo was significantly higher with the G/A and A/A genotypes compared with HCs (p = 0.011). Similarly, a significantly increased risk was noted in patients with A allele vs. G allele (p<0.0001). In AA and vitiligo patients, a significant increase in sTNF-α levels was noted in the mutant G/A genotypes vs. the normal G/G genotype (p<0.0001) and in the A allele vs the G allele (p<0.0001). According to the in silico analysis, this SNP could mainly affect the SP1 transcription factor binding site with subsequent effect on TNF-α expression. CONCLUSION: According to results of the laboratory and the in silico study, the mutant TNF-α (308) genotypes were risk factors that conferred susceptibility to vitiligo among Egyptian patients but had no effect on the susceptibility to AA.

Analysis of the expression profile of long non-coding RNAs MALAT1 and THRIL in children with immune thrombocytopenia.

BACKGROUND/AIMS: Pediatric immune thrombocytopenia (ITP) is an autoimmune disease; whose etiology is not exactly understood and seems to be highly multifactorial. Long non-coding RNAs (lncRNAs) are key regulators of different actions, which contribute to the development of many autoimmune diseases. To gain a further understanding, we estimated the relative expression of lncRNAs Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) and tumor necrosis factor-α (TNF-α) and heterogeneous nuclear ribonucleoprotein L (hnRNPL) immune-regulatory lncRNA (THRIL) in pediatric ITP. METHODS: In this case-control study, analysis of the expression profiles of these lncRNAs in blood samples from children with ITP and healthy controls (HCs) using quantitative real-time PCR was done. The association of MALAT1 and THRIL with ITP clinical features and their potential usage as non-invasive circulating biomarkers for ITP diagnosis was also evaluated. The receiver operating characteristic curve was constructed, and an area under the curve was analyzed. RESULTS: Both lncRNAs MALAT1 and THRIL were significantly upregulated in ITP patients in comparison to HCs ( p < .0001 and = .001 respectively). In addition, there was a positive significant correlation between the expression level of both biomarkers among patients (r = 0.745, p < .0001). At cutoff points of 1.17 and 1.27 for lncRNAs MALAT1and THRIL, respectively, both biomarkers had an excellent specificity (100% for both) and fair sensitivity (63.6 and 73.3% for lncRNAs MALAT1and THRIL, respectively). Improvement of biomarkers specificity was obtained by evaluation of the combined expression of both biomarkers. Serum lncRNAs MALAT1 and THRIL could be used as potential biomarkers in differentiating childhood ITP patients and HCs.

Chronic Hepatitis C Infection Has No Effect on Peripheral CD4+CD25+ Tregulatory Cells in Patients with End-Stage Renal Disease.

Background: T regulatory cells (Tregs), through variable mechanisms, play a crucial role in Hepatitis C virus (HCV) chronicity and infection tolerance. A great speculation is posed regarding the level, role of Tregs in end-stage renal disease (ESRD), and the presence of associated factors that could influence the Tregs population. Accordingly, we aimed at studying the effect of HCV infection on peripheral CD4+CD25+Tregs population among patients on hemodialysis (HD) as well as the effect of other comorbidities on these cells.Patients and methods: A group of 77 patients on HD (32 were HD HCV+ and 45 were HD HCV-) and 80 healthy controls (HCs) were included in the study. Flow cytometric analysis was performed for identification and quantification of peripheral CD4+ CD25+Tregs.Results: The frequency of CD4+ CD25+Tregs increased significantly in HD patients compared to the HCs (p = <.0001 each). HCV posed no effect on peripheral CD4+ CD25+ Tregs in ESRD patients, when comparing HD HCV- and HD HCV+ groups. In the hypertensive HD HCV-, Tregs percentage was higher than that in the non-hypertensive. However, the difference was not statistically significant. No significant difference was detected between HD HCV- and HD HCV+ patients on the count and percentages of Tregs according to the duration of dialysis.Conclusion: Demonstrating that chronic HCV infection has no effect on CD4+ CD25+ Tregs cells levels in ESRD patients is of great importance to the success of future allografts in such patients.

A Study of the Virulence Traits of Carbapenem-Resistant Klebsiella pneumoniae Isolates in a Galleria mellonella Model.

The increasing incidence of carbapenem-resistant Klebsiella pneumoniae (CRKP) strains is considered as a terrifying public health concern. This study target was to gain a further insight into the virulence traits of CRKP isolates in Egypt. The study was carried out by using 43 clinical K. pneumoniae isolates. Antibiotic susceptibility testing, biofilm formation assay, and molecular characterization of carbapenemase and virulence genes were done for all isolates. In addition, the genotypic relationship between CRKP isolates was identified by using enterobacterial repetitive intergenic consensus-polymerase chain reactions (ERIC-PCRs). A Galleria mellonella survival assay was adopted for in vivo testing of virulence of the CRKP. Carbapenem resistance was exhibited among 58% (25/43) isolates. Minimum inhibitory concentration values of carbapenem-resistant K. pneumoniae (CRKP) ranged from 32 to 128 µg/mL. Biofilm assay has revealed that 21 isolates (49%) had moderate biofilm formation and 11 isolates (25.5%) were strong biofilm producers. BlaNDM-1 was recognized in 20.9% (9/43) of the isolates, while blaOXA-48 was observed in 18.5% (8/43). Type 3 fimbriae (mrkD) and entB were addressed among 72.1% and 62.8% of K. pneumoniae isolates, respectively. The ybtS and iutA genes were detected among 44.2% and 37.2% of the isolates, respectively. ERIC-PCR showed 23 genetic profiles among CRKP isolates. CRKP biofilm producers were virulent according to the G. mellonella model, which indicates the importance of biofilm as a virulence trait among CRKP. This study indicates the emergence of CRKP with increased virulence traits, especially biofilm formation, in Egypt. This alarming report highlights the ongoing need for effective screening procedures and strict infection control measures.

Expression Profile of Long Noncoding RNAs, lnc-Cox2, and HOTAIR in Rheumatoid Arthritis Patients.

Despite the increased proof that long noncoding RNAs (lncRNAs) can control gene expression and broadly affect the normal physiological and disease conditions, the part of lncRNAs in rheumatoid arthritis (RA) is not well known. This study aimed to assess the serum expression levels of lnc-Cox2 and HOTAIR in RA and to investigate their role as novel noninvasive biomarkers in diagnosis of RA. Also, their relations with the levels of interleukin (IL)-6 and matrix metalloproteinase (MMP)-9 and with other clinicolaboratory data in RA patients were analyzed. LncRNAs-Cox2 and HOTAIR expression levels were detected in serum by real-time quantitative polymerase chain reaction. Both IL-6 and MMP-9 levels in serum were measured by enzyme-linked immunosorbent assay. The mRNA expression of lncRNA-Cox2 and HOTAIR was significantly upregulated in RA patients compared with healthy controls. Serum levels of both IL-6 and MMP-9 were significantly higher in RA patients than in healthy subjects (P < 0.001 each). Receiver operating characteristic (ROC) curve demonstrated that lncRNA-Cox2 and HOTAIR could discriminate RA patients from healthy controls. HOTAIR (not lnc-Cox2) was observed to be an independent predictor for RA using multiple logistic regression analysis. We concluded that lnc-Cox2 and HOTAIR serum expression levels can be used as novel noninvasive biomarkers for the diagnosis of RA.

In vivo activity of co-trimoxazole combined with colistin against Acinetobacter baumannii producing OXA-23 in a Galleria mellonella model.

OBJECTIVES: Acinetobacter baumannii is a critical nosocomial pathogen. A. baumannii infections have become a grave challenge due to their ability to develop resistance to different antimicrobial agents. The current study aimed to evaluate the potential synergism and bactericidal activity of a combination of colistin and cotrimoxazole against carbapenem-resistant A. baumannii (CRAB) in a Galleria mellonella model. METHODS: Four clinical A. baumannii isolates were biochemically and molecularly identified. Their antimicrobial susceptibility levels were established and the molecular characterization of the carbapenemase-encoding genes was performed. The synergism and bactericidal effect of the colistin/cotrimoxazole combination was assessed using the checkerboard assay and time-kill experiments. An in vivo evaluation of the activity of the combination was performed using the Galleria mellonella model. RESULTS: A fractional inhibitory concentration index (FICI) of ≤0.5 was found for all strains, indicating that the colistin/cotrimoxazole combination exhibited powerful synergistic activity. The combination displayed both synergistic and bactericidal activity at sub-breakpoint concentrations for all strains. Cotrimoxazole monotherapy showed the least protective activity in the G. mellonella model. The survival rate ranged from 66.7-79.2 % at 24 h and was 29.2-60.4 % at 96 h for the tested isolates. Colistin monotherapy performed better than cotrimoxazole monotherapy; the G. mellonella survival rate ranged from 77.1-97.9 %, at 24 h and from 64.5-72. % at 96 h. The colistin/cotrimoxazole combination improved G. mellonella's survival rate at 96 h remarkably in comparison to colistin or cotrimoxazole monotherapy. CONCLUSIONS: Finally, the combination of colistin and cotrimoxazole appears to be a promising therapeutic option for the management of CRAB-associated infections. It is essential to assess the clinical application and the dose-response relationships of combinations such as colistin plus cotrimoxazole.

Interplay Between Helicobacter pylori Infection, Interleukin-11, and Leukemia Inhibitory Factor in Gastric Cancer Among Egyptian Patients.

Helicobacter pylori is a ubiquitous Gram-negative bacterium, that is responsible for gastric mucosal inflammation. It is the most common risk factor for gastric cancer (GC). The current study aimed to investigate the association between interleukin-11 (IL-11) and leukemia inhibitory factor (LIF) levels among H. pylori-infected Egyptian patients with gastritis and GC. One hundred forty-seven patients with gastric lesions were endoscopically biopsied and assessed using rapid urease test and immunohistochemistry. Quantitative real-time polymerase chain reaction was done for the detection of H. pylori load in gastric biopsies and detection of LIF as well as IL-11 relative gene expression. The mean values of H. pylori load, LIF, and IL-11 were significantly elevated in GC patients compared to gastritis group (P < 0.0001). A positive significant correlation was detected between mucosal levels of LIF, IL-11, and H. pylori load in both groups. Both LIF and IL-11 had the same pattern of expression in gastric tissues with different types of gastritis and different types and grades of gastric carcinoma. This report could clarify the molecular events associated with the immune response against H. pylori infection and H. pylori-associated pathology. Therefore, development of immunotherapy strategies against H. pylori-induced cytokines becomes inevitable.

Hospital outpatient clinics as a potential hazard for healthcare associated infections.

Healthcare acquired infections are no longer confined to the hospital environment. Recently, many reported outbreaks have been linked to outpatient settings and attributed to non-adherence to recommended infection-prevention procedures. This study was divided into two parts: The first is a descriptive cross-sectional part, to assess the healthcare personnel's knowledge and compliance with Standard Precautions (SP). The second is an intervention part to assess the role of health education on reducing the level of environmental and reusable medical equipment bacterial contamination. Assessment of the doctors' and nurses' knowledge and compliance with SP was performed using a self-administered questionnaire. Assessment of environmental cleaning (EC) and reusable medical equipment disinfection has been performed using aseptic swabbing method. The extent of any growth was recorded according to the suggested standards: (A) Presence of indicator organisms, with the proposed standard being <1cfu/cm(2). These include Staphylococcus aureus (including methicillin-resistant Staphylococcus aureus, MRSA), Enterococci, including vancomycin-resistant Enterococci (VRE) and various multidrug-resistant Gram-negative bacilli. (B) Aerobic colony count, the suggested standard is <5cfu/cm(2). The effect of health education intervention on cleaning and disinfection had been analyzed by comparing the difference in cleaning level before and after interventional education. Good knowledge and compliance scores were found in more than 50% of participants. Primary screening found poor EC and equipment disinfection as 67% and 83.3% of stethoscopes and ultrasound transducers, respectively, were contaminated with indicator organisms. For all indicator organisms, a significant reduction was detected after intervention (p=0.00). Prevalence of MRSA was 38.9% and 16.7%, of the total S. aureus isolates, before and after intervention, respectively. Although 27.8% of the total Enterococcus isolates were VRE before intervention, no VRE isolates were detected after intervention. These differences were significant. Development and monitoring of the implementation of infection prevention policies and training of HCP is recommended.

The relationship between antisperm antibodies prevalence and genital chlamydia trachomatis infection in women with unexplained infertility.

Chlamydia trachomatis infection is one of the most common sexually transmitted diseases and sperm-associated antibody could impair fertility through various mechanisms. Both factors could be correlated to affect the fertility status of women. A retrospective case-control study was performed enrolling ninety (n = 90) patients with primary or secondary infertility as the case group, in addition to another eighty (n = 80) healthy women attending the family planning clinic to investigate the correlation between C. trachomatis past and current infections and antisperm antibodies (ASA) in women with unexplained infertility. The PCR prevalence of C. trachomatis didn't differ significantly among both groups (2.4 versus 1.6%, P = 0.66). In contrast, significantly higher prevalence of anti-C. trachomatis specific IgG (39% versus 19%, P = 0.87) antibodies were found among infertile women. ASA prevalence was significantly higher in infertile group (20 % versus 5%, P = 0.04). The final study results have failed to find a positive correlation between current or past C. trachomatis infection and the level of antisperm antibodies level in women suffering of un-explained infertility. Anti-sperm antibodies were significantly higher in infertile women, but without a significant difference between the incidences of ASA in infertile women with past or current C. trachomatis current infection.