Burden of pediatric influenza A virus infection post swine-flu H1N1pandemic in Egypt

Objectne:To screen children with influenza like illness or with symptoms of acute respiratory tract infections for influenza A virus infection-post swine flu pandemic era-using rapid influenza diagnostic tests.Methods:During two year,(2010&2011),1200 children with influenza like illness or acute respiratory tract infections(according to World Health Organization criteria)were recruited.Their ages ranged from 2-60 months.Nasopharyngeal aspirates specimens were collected from all children for rapid influenza A diagnostic test.Results:Influenza A virus rapid test was positive in 47.5%of the children;the majority(89.6%)were presented with lower respiratory tract infections.Respiratory rate and temperature were significantly higher among positive rapid influenza test patients.Conclusions:Influenza A virus infection is still a major cause of respiratory tract infections in Egyptian children.It should be considered in all cases with cough and febrile episodes and influenza like symptoms even post swine flu pandemic.

<i>In Vitro</i>Activity of Colistin and Vancomycin or Azithromycin Combinations on Extensively Drug Resistant <i>Acinetobacter baumannii</i>Clinical Isolates

Background: Extensively drug resistant Acinetobacter baumannii (XDR-AB) presents an increasing challenge to health care in Egypt as they are among the most common bacteria isolated in hospital setting. Treatment of such infections usually involves the use of antimicrobial agents in combination. Various combinations have been proposed, with colistin serving as the backbone in many of them even for colistin resistant isolates. Aim: The study was conducted in order to test the in vitro combined effects of colistin and vancomycin or azithromycin against (XDR-AB) causing infections at Alexandria Main University Hospital in Egypt, in an attempt to detect the possibility of a beneficial combination therapy. Material/Methods: Thirty XDR-AB clinical isolates were included in the study. Antibiotic susceptibility testing was performed using automated Vitek 2 compact system and disc diffusion method. Colistin antibiotic disc diffusion test was compared with broth microdilution method. Organisms were also tested against colistin and vancomycin or azithromycin in combination using checkerboard synergy test and FICI (Fractional Inhibitory Concentration Index) was calculated. Synergy was defined as a FICI of ≤0.5. Results: On comparing the two methods used to detect susceptibility to colistin to broth microdilution for MIC (minimum inhibitory concentration) determination, as a reference method, the Vitek showed 100% categorical agreement (CA), on the other hand, the disc diffusion showed CA of 93% with very major errors. Synergy was detected for all isolates (100%) when combining colistin with vancomycin (FICI mean = 0.08). As for azithromycin, 21 strains had FICI range from 0.7 to 1.001, denoting indifference;the remaining 9 strains showed synergy with FICI range from 0.06 to 0.241. The mean colistin/azithromycin FICI was 0.71 for the 30 isolates. Conclusion: These findings suggest that regimens containing vancomycin may confer therapeutic benefit for infection due to XDR-AB;however, other methods (time-kill assay) should be used to confirm such synergy. Furthermore, the optimal combination treatment for serious XDR-AB infection should be addressed in a prospective clinical trial.

MBL2 Gene Polymorphism and the Association with Neonatal Sepsis in Egyptian Neonates, a Case Control Study

Mannose binding lectin (MBL) is an important component of innate immunity particularly in neonates whose adaptive immunity is not fully developed. Polymorphism in MBL2 gene promoter and exon1 determines MBL serum level and function. The aim of this study was to investigate the frequency of different MBL2 genotypes in neonatal sepsis among patients of neonatal intensive care unit (NICU). Two hundred and forty-five neonates were enrolled in this study (127 infected and 118 uninfected controls). Multiplex PCR and double amplification refractory mutation system (dARMS) were used for typing of MBL2 exon1 and promoter respectively. Klebsiella species were the most frequently isolated organisms (22.8%). There is no statistical significance difference in the distribution of different expression genotypes between infected group and controls (P = 0.11). However, prevalence of low MBL2 expression genotypes (XA/O and O/O) was higher in infected patients compared to control group (patients 25.2% and controls 15.3%). Low and medium MBL2 expression genotypes were mostly associated with Gram-negative bacterial infections (18.9% and 22.8%) respectively. A statistically significant association of Gram-negative bacterial infections with low MBL2 expression genotypes was found (P = 0.02). Higher frequency of AB and BB genotypes was observed (31.5% and 7.9%) in patients group compared to control, but without statistical significant difference.

The Role of Circulating MicroRNAs as Markers of Disease Progression in Hepatitis C Virus Infected Egyptian Patients

Background: The discovery of miRNAs circulating in the peripheral blood has opened new directions of research to identify new non-invasive markers for diagnosis of diseases. Aim: The aim of the study was to evaluate the expression levels of circulating plasma miRNAs (miRNA-21 & miRNA-122) in Egyptian patients with chronic uncomplicated and complicated HCV. Patients & Methods: This study was conducted on 60 Chronic HCV infected patients. Patients were divided into three groups (20 patients each): uncomplicated HCV, cirrhosis, and hepatocellular carcinoma (HCC). All patients were subjected to laboratory investigations including complete blood picture, liver function tests. Expression levels of miRNA-21 and -122 in plasma using RT-PCR were determined. Results: MiRNA-21 showed significant fold increase in chronic uncomplicated HCV while significant fold decrease in cirrhotic and HCC groups (P = 0.036). On the other hand, miRNA-122 showed significant fold elevation in both chronic uncomplicated and cirrhotic groups and significant fold decrease in HCC group (P = 0.005). ROC curve analysis for miRNA-122 yielded 68.4% sensitivity and 100% specificity for the differentiation of HCC patients from non-HCC at a cutoff 0.184. Neither miRNA-21 nor miRNA-122 was a successful predictor for HCC diagnosis. Conclusion: MiRNA-122 can be used as novel non-invasive biomarker for monitoring HCV related disease progression.

Tier-based approach to establish a culture of biosafety at a medical microbiology research laboratory in Egypt

Microbiology Research Laboratory(MRL)is a biosafety level-2(BSL-2)research laboratory located at the main campus of Faculty of Medicine,Ain Shams University(ASU)in Cairo.With the objective of strengthening the departmental capacities of biosafety,a series of activities were carried out between October 2019,and January 2020 to raise awareness,along with instilling standard biosafety practices and procedures among laboratory staff including non-health professions.MRL staff were categorized according to their biosafety knowledge into three tiers:tier(1):with zero to minimal knowledge,tier(2):with basic knowledge,tier(3):with satisfactory knowledge.Tier based activities were designed to align with their job responsibilities.Results:44 selected laboratory staff were trained on biosafety practices:12 from tier(1),19 from tier(2)and 13 constituted tier(3).Through regular follow-ups,the impact of the implemnted training plan was reflected on the practices and knowledge of all laboratory staff.Knowledge among health professions has increased by 60%.Furthermore,6 staff members have granted a biosafety certification by International Federation of Biosafety Association(IFBSA).Conclusion:establishing a culture of biosafety within microbiology research laboratories is integral to safe research practices.Together with developing local and national biosafety regulations and policies will ensure research advancement without compromising public health or environmental safety.

Assessing decontamination practices at a medical microbiology research laboratory

To our knowledge,this is the first study to conduct an objective assessment of the routine decontamination practices at a medical microbiology research laboratory(MRL)a year after a biosafety training was provided to all laboratory staff.Between March 28th and June 28th,2021,unobtrusive observations were carried out to identify-three high-touch surfaces at the MRL during different working hours.Swabbing was used to evaluate the effectiveness of the disinfectant used in the laboratory.All three high-touch surfaces were sampled before and after decontamination with 200 ppm of 5%sodium hypochlorite(household bleach)to quantify the microbial load and identify the types of organisms residing on the laboratory surfaces.A higher concentration(500 ppm)of 5%sodium hypochlorite was employed after refresher training was provided to housekeeping staff,and resampling of the three surfaces was carried out during a 4-week follow-up period using the same procedure.The three high-touch surfaces identified were the two sides of the workbench(22%–24%)and the front surface of one incubator(14%).Anthracoid bacilli and Staphylococcus aureus were the most commonly found organisms on laboratory surfaces preintervention(100%and 89%,respectively)and post-intervention(56%and 44%,respectively).Other microorganisms detected included Salmonella spp.(27.7%),Proteus spp.(5.6%),Escherichia coli(5.6%),and Klebsiella spp.(33.3%).Employing a higher concentration(500 ppm)of sodium hypochlorite significantly(p<0.000)reduced the total aerobic colony count from an average of 15–250 cfu/cm2 to 10–60 cfu/cm^(2).This study demonstrated suboptimal decontamination practices at the MRL and the need to apply a higher concentration(500 ppm)of sodium hypochlorite to reduce the overall microbial load.It also demonstrated the importance of quantitative assessment to monitor decontamination practices and ensure staff compliance.More studies are needed to identify bacterial communities within the laboratory,which will help provide guidance regarding the types,proper concentrations,and appropriateness of the in-use disinfectants.Furthermore,large-scale studies on the acceptable level of residual contamination following any decontamination process are urgently recommended.

Activity of Fosfomycin in Extended-Spectrum Beta-Lactamases Producing Klebsiella pneumonae from Hospital Acquired Urinary Tract Infections

Treatment of hospital acquired urinary tract infections (UTIs) caused by extended-spectrum beta-Lactamases producing Klebsiella pneumonae is a major problem. This organism expresses a high level of resistance to many groups of antibiotics. Fosfomycin is an agent which is recommended for treatment of UTIs caused by ESBLs producers. The aim of this study is to determine the sensitivity pattern of ESBLs producing urinary K. pneumonae to antimicrobial agents including fosfomycin in patients of MUHs and determine the prevalence of fosfomycin resistance mediated by plasmid mediated fosfomycin modifying enzymes fosA, fosB and fosA3. Methods: Klebsiella pneumonae urinary isolates were collected from patients with hospital acquired UTIs in Mansoura University Hospitals (MUHs). The susceptibility pattern was determined by Kirby Baur method. Isolates resistant to extended spectrum cephalosporins were tested for ESBLs production by double disc diffusion method. Fosfomycin resistance was determined by broth dilution method. Isolates resistant to fosfomycin were tested for fosA, fosB and fosA3 by PCR. Results: A total of 128 ESBLs producing K. pneumonae isolates were collected. The highest sensitivity was to imipenem (94.5%). The lowest was to trimethoprime-sulphamethoxazole (21.8%). Co-resistance of ESBLs isolates with fosfomycin was 23.2%. Eighteen fosfomycin resistant isolates (18/30) were positive to fosA. Conclusion: ESBLs producing urinary Klebsiella pneumonae express moderate sensitivity to fosfomycin. Resistance is mainly mediated by plasmid mediated fosfomycin modifying enzymes fosA.

Maternal vitamin D level and vitamin D receptor gene polymorphism as a risk factor for congenital heart diseases in offspring;An Egyptian case-control study

Vitamin D&vitamin D receptor(VDR)signaling play a very crucial role in early embryonic heart development.We construct this case-control study to investigate the association between maternal serum vitamin D level&VDR gene Fok1 polymorphism and risk of congenital heart defects(CHD)in offspring.Fifty mothers who had term neonates with CHD were considered as cases.Fifty age-comparable healthy mothers who had neonates without CHD were contemplated as controls.Maternal serum 25 hydroxyvitamin D[25(OH)D]level was tested using ELISA.Maternal VDR gene Fok1 polymorphism was analyzed using PCR-based RFLP-assay.There was a significant decrease in maternal vitamin D level(P=0.002)and a significant increase in vitamin D deficient status(P=0.007)among cases when compared to controls.VDR gene Fok1 genotypes distribution frequency were in accordance with Hardy Weinberg equilibrium(HW)among controls.A significant increase in VDR gene Fok1 F/f&f/f genotypes and f allele were observed in cases compared to controls with estimated odds ratio(95%confidence interval)&P-value of 3(1e8)&P=0.006,11(1e97)&P=0.01 and 3(2e6)&P=0.001 respectively.There was a significant decrease in maternal vitamin D level in neonates with cyanotic CHD(P=0.000)compared to those with a cyanotic CHD while there was no significant difference in VDR Fok1 genotype(P=0.18)&allele(P=0.05)distribution between two groups.We concluded that maternal vitamin D deficiency and VDR gene Fok1 F/f,f/f genotype and f allele were associated with increased risk of CHD in offspring.

Expansion of GARP-Expressing CD4^(+)CD25^(-)FoxP3^(+)T Cells and SATB1Association with Activation and Coagulation in Immune Compromised HIV-1-Infected Individuals in South Africa

Although antiretroviral treatment lowers the burden of human immunodeficiency virus(HIV)-related disease,it does not always result in immunological recovery.This manifests as persistent chronic inflammation,immune activation or exhaustion that can promote the onset of co-morbidities.As the exact function of regulatory T(Treg)cells in HIV remains unclear,this cross-sectional study investigated three expression markers(Forkhead box protein P3[FOXP3],glycoprotein A repetitions predominant[GARP],special AT-rich sequence binding protein 1[SATB1])and compared their expansion between CD4^(+)CD25^(-)and CD4^(+)CD25^(++)T cells.Age-matched study subjects were recruited(Western Cape,South Africa)and sub-divided:HIV-negative subjects(n=12),HIV-positive na(i|")ve treated(n=22),HIV-positive treated based on CD4 count cells/μL(CD4>500 and CD4<500)(n=34)and HIV-treated based on viral load(VL)copies/mL(VL<1000 and VL>1000)(n=34).Markers of immune activation(CD38)and coagulation(CD142)on T cells(CD8)were assessed by flow cytometry together with FOXP3,GARP and SATB1 expression on CD4^(+)CD25^(-)and CD4^(+)CD25^(++)T cells.Plasma levels of interleukin-10(IL-10;anti-inflammatory marker),IL-6(inflammatory marker)and D-dimer(coagulation marker)were assessed.This study revealed three major findings in immuno-compromised patients with virological failure(CD4<500;VL>1000):(1)the expansion of the unconventional Treg cell subset(CD4^(+)CD25^(-)FOXP3^(+))is linked with disease progression markers;(2)increased GARP expression in the CD4^(+)CD25^(-)and CD4^(+)CD25^(++)subsets;and(3)the identification of a strong link between CD4^(+)CD25^(-)SATB1+cells and markers of immune activation(CD8^(+)CD38^(+))and coagulation(CD8^(+)CD142^(+)and D-dimer).