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Introduction: Collagens are the most abundant proteins in the human body, accounting for one-third of total proteins. Over the last few years, accumulated evidence have indicated that some collagens are differentially expressed in cancer. The aim of the study was to assess COL1A1 gene expression as a novel marker for the progression of hepatitis B cirrhosis into hepatocellular carcinoma. Methods: This cohort study included 348 subjects and was conducted between May 2018 and June 2019. Subjects were divided into 4 groups: group1 included HBV positive hepatocellular carcinoma patients “HCC” (n= 87), group II included HBV positive patients with liver cirrhosis “LC” (n = 87), group III included chronic hepatitis B patients with neither HCC nor cirrhosis “ C-HBV” (n = 87) and group IV consisted of healthy volunteers as controls (n = 87). Fasting venous blood samples (10 ml) were collected from each participant in this study and were used for assessment of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin, albumin and alfa-fetoprotein (AFP). Another portion of blood was collected in 2 vacutainer tubes containing EDTA, one for Complete blood count and the other for gene expression of COL1A1. Results: The gene expression of collagen was 6.9 ± 8.8 in group 1 (HBV positive hepatocellular carcinoma patients) and this was a significant increase in comparison with the other groups. In group 2 (HBV positive patients with liver cirrhosis), the gene expression (collagen) was 3.7±1.5 and it was significantly increased when compared with group 4 (healthy volunteers). Conclusion: COL1A1 gene expression can be used as an indicator of the progression of hepatitis B cirrhosis into hepatocellular carcinoma.

Background and Objectives: Dental caries is a breakdown of the teeth enamel due to harmful bacteria, lack of oral hygiene, and sugar consumption. The acid-producing bacterium Streptococcus mutans is the leading cause of dental caries. Dextra- nase is an enzyme that can degrade dextran to low molecular weight fractions, which have many therapeutic and industrial applications. The purpose of the present study was to isolate a novel dextranase-producing bacteria from asource (molasses). The cell-free extracts containing dextranases were tested as antibiofilm agents. Materials and Methods: Dextranase-producing bacteria were identified using phenotypic and genotypic methods such as 16S rRNA gene sequencing and enzymatic characterization. Results: The highest six dextranase-producing bacterial isolates were Bacillus species. The best conditions for dextranase productivity were obtained after 72hours of culture time at pH 7. The addition of glucose to the medium enhanced the pro- duction of the enzymes. The cell-free extract of the six most activeisolates showed remarkable activity against biofilm for- mation by Streptococcus mutans ATCC 25175. The highest inhibition activities reached 60% and 80%for Bacillus velezensis and Pseudomonas stutzeri, respectively. Conclusion: Therefore, our study added to the current dextranase-producing bacteria with potential as a source of dextra- nases.

The biological fitness cost of antibiotic resistance is a key parameter in determining the rate of appearance and spread of antibiotic-resistant bacteria in Egypt. Our study aimed to investigate the prevalence of antibiotic resistance among Escherichia coli clinical isolates from Greater Cairo area hospitals. A total of 537 clinical isolates were recovered from samples of urine, diarrheal specimen, pus, wound culture, gastric wound, blood, drain culture, sputum, high vaginal swab, abscess, amniotic fluid, ventilator, burn swab, splenic drain culture, and unknown site of infection during different seasons. All isolates were subjected to phenotypic and genotypic susceptibility testing for colistin, nitrofurantoin, fosfomycin, and trimethoprim, quinolones, and β-lactam resistance. Our results revealed that 42.7% of the isolates harbored at least one resistance encoding gene, 10% harboring 2, 0.6% harboring 3, and 0.85% harboring 4 resistance-encoding genes. PCR reported the prevalence of resistance genes as follows: bla-SHV 13.4%, mcr-1 0.6%, qnr-A 23.8%, fos-A 1.06%, nfs-A 3.6%, and dfr-A 25.5%. We reported that three isolates carried the mcr-1 gene encoding colistin resistance from three different hospitals. Upon performing sequencing and phylogenetic analysis on the three positive mcr-1 isolates (MT890587, MT890588, and MT890589), the three isolates showed 100% identity with themselves, with some strains from Egypt and Japan, and 99.9% identity with an isolate from China.

Background: Polymorphisms of genes encoding the pro-inflammatory and anti-inflammatory cytokines can affect the clinical presentation of the infection. We aimed to assess the role of EGF gene single-nucleotide polymorphism in the outcome of chronic hepatitis B virus (HBV) infection in children. Methods: One hundred HBV-infected children and 75 healthy matched controls were enrolled in this prospective study. Patients included 18 chronic inactive and 82 chronic active carriers. EGF rs4444903 A>G genotypes were determined using allele-specific amplification. Results: Significant differences regarding EGF genotypic frequency (p=0.001) in patients compared to controls (p=0.001). Eighteen percent were inactive, and 82% were active carriers. AA, AG and GG genotypic frequency were 66.7%, 33.3%, 0% and were 3.7%, 37.8% and 58.5% in the inactive and active carriers, respectively, with significant differences regarding AA, AG, GG genotypic frequency (p=0.001 for all). EGF AA, AG, GG genotypes frequency were 1.9%, 33.3%, and 64.8%, respectively, with significant differences between cirrhotic and non-cirrhotic patients regarding AA, AG, GG genotypic frequency (p=0.001 for all). Conclusion: Increased G allele frequency in EGF rs4444903 A > G polymorphism in HBV- Egyptian children is associated with worse outcomes.

Background/Aim: Spontaneous bacterial peritonitis is a common bacterial infection of ascitic fluid mainly in ascites due to liver cirrhosis. Mannose-binding lectin (MBL) can activate phagocytosis and the complement system. Spontaneous bacterial peritonitis was detected to be higher in MBL deficiency. This study aimed to assess ascitic fluid MBL in liver cirrhosis and spontaneous bacterial peritonitis. Methods: Ninety patients with cirrhotic ascites were included. Forty five of them had SBP. Child-Pugh score, Model for End Stage Liver Disease (MELD) and its update (uMELD) scores were used to assess severity of liver cirrhosis. Ascitic fluid samples were obtained for differentiation of leucocytic count, estimation of albumin, protein, glucose, and serumascitic albumin gradient. Ascitic fluid levels of MBL were measured for all patients. SBP was documented if polymorphonuclear leucocytic count ≥250/mm in ascitic fluid. Results: Ascitic fluid MBL level was significantly lower in patients with SBP. MBL had a significant negative correlation with ascitic total leukocytic count (TLC), also with serum creatinine, bilirubin, PT, INR and MELD score among SBP patients. However, it had a significant positive correlation with ascitic protein and with platelets. According to multivariate analysis; fever, TLC, platelets, creatinine, MBL, glucose and polymorphs were independent predictors for SBP development. Conclusion: Ascitic fluid MBL could be a good predictive and prognostic marker in patients with cirrhosis and spontaneous bacterial peritonitis.

AIM: To detect the quantitative expression levels of the pro-inflammatory interleukin-8 (IL8), antimicrobial peptides human beta defense-2 (HBD2), and human beta defense-3 (HBD3) genes in bacterial conjunctivitis. METHODS: The human conjunctival epithelial cells were obtained using the impression cytology technique from healthy controls and patients. The genes expression levels were determined utilizing a reverse transcription quantitative polymerase chain reaction (RT-qPCR). The contribution of causative agent type, the number of isolates and severity of clinical features, in the increase of genes expression was also determined. RESULTS: The RT-qPCR showed that IL8, HBD2, and HBD3 expression increased in bacterial conjunctivitis as compared to healthy control (P

Background & Aims: The aim of the study was to evaluate serum c-reactive protein (CRP), ascitic procalcitonin (PCT) and monocyte chemotactic protein-1 (MCP-1) in the diagnosis of spontaneous bacterial peritonitis (SBP) in cirrhotic patients. Methods: A cross-sectional analytic study that included 199 patients with decompensated cirrhosis (101 with SBP and 98 without SBP). Patients were classified according to Child-Pugh criteria. Ascitic PCT and MCP-1 were measured by enzyme-linked immunosorbent assay. Serum CRP, liver and renal functions were assessed. Results: Three markers are significantly elevated in SBP patients than those without ascites. Using the ROC curve at AUC 0.883 and a cut-off value of >186 ng/ml, the diagnostic performance of ascitic MCP-1 level was higher than CRP (AUC 0.562) and ascitic fluid procalcitonin (AUC 0.751) in the diagnosis of SBP. The sensitivity and specificity were 86.15% and 79.59% at the cutoff of 186 ng/ml for MCP-1, 65.4 and 75.5 at ≥ 1 ng/ml for PCT, and 52.5 and 64.3, respectively for at 11.2 mg/dl CRP. Conclusion: Ascitic MCP-1 has a better diagnostic value with higher sensitivity and specificity in diagnosis SBP compared to CRP and procalcitonin which has higher diagnostic accuracy than CRP. Further studies with a large number will be necessary to evaluate the usefulness of these markers in diagnosis, follow-up and relation to morbidity and mortality of SBP patients.

Ciprofloxacin (CIP) and levofloxacin (LEV), widely used fluoroquinolone antibiotics, are often found in sewage from the sewage treatment plants and marine environment. In this study, CIP and LEV biodegrading bacterial consortia were obtained from industrial wastewater. Microorganisms in these consortia were identified as Acinetobacter baumannii (A. baumannii), Klebsiella pneumoniae (K. pneumoniae) and Elizabethkingia miricola (E. miricola). The impacts of the critical operating parameters on the elimination of CIP and LEV by bacterial consortia have been investigated and optimized to achieve the maximum levels of CIP and LEV biodegradation. Using liquid chromatography with tandem mass spectrometry (LC-MS-MS), possible degradation pathways for CIP and LEV were suggested by analyzing the intermediate degradation products. The role of the enzymes fluoroquinolone-acetylating aminoglycoside (6′-N-acetyltransferase) and cytochrome P450 (CYP450) in the breakdown of fluoroquinolones (FQs) was investigated as well. According to our findings, various biodegradation mechanisms have been suggested, including cleavage of piperazine ring, substitution of F atom, hydroxylation, decarboxylation, and acetylation, as the main biotransformation reactions. This study discovers the ability of non-reported bacterial strains to biodegrade both CIP and LEV as a sole carbon source, providing new insights into the biodegradation of CIP and LEV. Graphical abstract

AIM: To investigate and analyze the changes in the conjunctival flora of patients exposed to repeated antibiotic usage. In addition, determining the effects of this repeated exposure on the resistance pattern of the conjunctival flora. METHODS: This study included 40 patients, admitted to the Retina Unit of the Research Institute of Ophthalmology. All chosen patients had a minimum of 4 consecutive, monthly intravitreal injections (IVI), some were extended to 6mo or 1y as required. The patients were randomly divided into 3 groups and each group received one kind of antibiotic which was either ofloxacin, moxifloxacin or ceftazidime. All bacterial isolates were tested for antibiotic susceptibility using Kirby-Bauer disc diffusion technique. RESULTS: In this study the conjunctival normal flora at base line culture varied from a predominance of Staphylococcus epidermidis (51.2%), followed by Staphylococcus aureus 14% to Micrococcus species 12.8% and other coagulase negative staphylococci (CONS) 13%. Our results showed an increase in the percentage of Staphylococcus epidermidis among ceftazidime treated eyes during the four visits in comparison to baseline cultures of patients randomized to ceftazidime. In contrast, there was no noticeable increase in Staphylococcus aureus percentage from baseline. In fluoroquinolone treated eyes, we also observed an increase in percentage of Staphylococcus epidermidis from base line. In contrast to ceftazidime treated eyes, the Staphylococcus aureus percentage in fluoroquinolone treated eyes showed an increase from baseline. However, the pattern of the ocular flora composition changed with the exposure to the old and newer generation of fluoroquinolones. We noticed an increase of Staphylococcus epidermidis in moxifloxacin treated eyes than that in ofloxacin treated eyes from baseline. There was no observed difference in the pattern of Staphylococcus aureus regarding exposure to older and newer generations of fluoroquinolones. CONCLUSION: Repeated use of ophthalmic antibiotics not only alters the composition of the normal ocular flora, but also selects for resistant strains.

In this study, we investigated the seroprevalence of anti-hepatitis D virus (HDV) antibodies in hepatitis B surface antigen (HBsAg)-positive children after 25 years of obligatory vaccination of infants against hepatitis B virus. This cross-sectional study included 120 treatment-naïve HBsAg-positive children, with a male-to-female ratio of 1.8:1 and a mean age of 7.8 ± 3.8 years (range, 1-17 years). Mothers were positive for HBsAg in 96.6% of the cases. HBeAg-positive chronic infection was observed in 60% of the cases, HBeAg-positive chronic hepatitis in 12.5%, and HBeAg-negative chronic infection in 26.7%. Anti-HDV antibodies were not detected in any of the cases. Thus, there is a lack of anti-HDV antibodies in HBsAg-positive children, despite the current burden in adults.

There are increasing demands for biogenic synthesis of silver nanoparticles (AgNPs) due to their unique properties and their numerous applications especially in biomedical field. Direct sunlight irradiation on aqueous garlic extract (AGE) represent fast, applicable, non-toxic, cheap, and ecofriendly method for (AgNPs) synthesis, with potentials to compete multi-drug-resistant (MDR) and biofilm-forming pathogens causing otitis media (OM). The aim of this study was to establish an eco-friendly method for synthesis of AgNPs, using aqueous garlic extract and sunlight, and moreover, detect its potential to inhibit MDR OM microbes. Obtained AgNPs were characterized by UV-visible spectral analysis, TEM, DLS, XRD, and FTIR. The effect of biosynthesized AgNPs on microbial growth, as well as biofilm productivity, was assessed against five resistant OM-causing strains, namely Bacillus cereus, Pseudomonas aeruginosa, Penicillium chrysogenum, Aspergillus niger, and Aspergillus flavus. Kirby-Bauer disc diffusion method was used to assess the inhibitory effect of 100, 50, 25, 12.5, and 6.25 μg/ml of AgNPs compared with tetracycline or nystatin and AGE. Additionally, the effect of 25 μg/ml of AgNPs on biofilm productivity of such strains was qualitatively assessed using Congo red agar (CRA) and TEM used to capture the changes inside the selected strains. The obtained AgNPs showed a highly significant (p value

Background & Aims: Several studies, in different populations, have focused on the role of HLA-DQ gene polymorphism in the pathogenesis of HBV infection. However, these findings are still controversial. This study aimed to determine HLA-DQ polymorphism in Chronic HBV patients and its impact on the response to antiviral therapy. Methods: This study was carried out on a total number of 188 participants, they were subdivided as follows: Group I (patients’ group): included 97 patients with chronic hepatitis B viral infection that was further subdivided according to response to treatment into responder and non-responder subgroups, Group II (Control group): included 91 normal healthy subjects who were matched to the patient group by sex and age. PCR (Polymerase Chain Reaction) testing, for HBV-DNA, was done for all participants enrolled in the study to measure the viral virus load before and after treatment. HLA- DQ polymorphism allelic discrimination assay was assayed using the Real-time equipment. Results: In a general analysis for the SNP rs7453920, the overall genotypes frequencies were 37% for A/A, 60.6% for A/G, and 37% for G/G. The G alleles of HLA-DQ rs7453920 were significantly increased in chronic HBV infection patients. A total of 77 (79.4%) patients were responders. Among this group, 72.7% were male, and the average age was 38.59 ±9.15 years. On evaluation of the association between polymorphisms in HLA-DQ gene and treatment response, the results indicated that response to treatment declined when patients were carrying the more unfavorable rs 7453920 GG with a response rate of 64%. Patients carrying the mutant allele AG, or the wild type allele AA were more likely to achieve a higher rate of response (84.8% and 83.3%, respectively). Conclusion: The presence of HLA-DQB2 rs 7453920-G serves as a risk factor for chronic HBV infection and treatment failure in the Egyptian population.

Background and Objective Hepcidin is the key regulator of iron metabolism and is a significant biomarker for systemic inflammatory states. Vitamin D is a powerful immunomodulator and plays a significant role in the inflammatory responses and fibrosis occurring due to hepatitis C virus (HCV) infection. This study assessed the level of vitamin D and serum hepcidin and its expression in peripheral blood of children with chronic hepatitis C (CHC) and correlated them with other serum markers to reflect iron metabolism and liver disease severity. Methods A total of 100 children were included in this study: 50 with HCV infection and 50 healthy controls. Biochemical parameters together with vitamin D, hepcidin, and its expression were all measured. Results The level of hepcidin and its expression together with vitamin D and hepcidin-to-ferritin (H/F) ratios were significantly reduced in patients, but the iron and ferritin levels were higher (Pr = 0.43, 0.31, 0.34, 0.28, and 0.91, respectively) but significant negative correlation with vitamin D (r = −0.37). Both hepcidin and ferritin were higher in patients with Child Pugh scores B and C than those with score A (P Conclusion Measuring serum hepcidin and its expression together with vitamin D levels in patients may have a prognostic value and is promising in the follow-up of the severity of liver disease.

The biological fitness cost of antibiotic resistance is a key parameter in determining the rate of appearance and spread of antibiotic-resistant bacteria in Egypt. Our study aimed to investigate the prevalence of antibiotic resistance among

The 2019 coronavirus pandemic (COVID-19) continues to expand worldwide. Although the number of cases and the death rate among children and adolescents are reported to be low compared to adults, limited data have been reported. We urgently need to find the treatment and vaccine to stop the epidemic. Vaccine development is in progress, but any approved and effective vaccine for COVID-19 will take at least 12 to 18 months. The World Health Organization (WHO), the Center for Disease Control and Prevention (CDC), and the Food and Agriculture Organization (FAO) have issued instructions and strategies for controlling COVID-19 outbreak to the general public, physicians, travelers and injured patients to follow so that the transmission to a healthy population can be prevented. In this review, we summarize demographic data, clinical characteristics, complications and outcomes and finally prevention and control strategies for this serious pandemic.

Methicillin-resistant Staphylococcus aureus (MRSA) continues to be a global problem in infection control. The highest proportions of MRSA are reported by Jordan, Egypt and Cyprus investigators, where more than 50% of the invasive isolates are methicillin-resistant. The aim of this work was to study the prevalence, antibiotic sensitivity and genetic discrimination of MRSA in Egypt. Microbiological identification was done using Gram stain, catalase, coagulase and mannitol fermentation along with biochemical identification by analytical profile index (API) tests. Molecular identification was conducted by the polymerase chain reaction (PCR) targeting 16S ribosomal RNA and the nuc genes. Additionally, identification of methicillin-resistant S. aureus (MRSA) was performed by the amplification of 310 bp of the mecA gene. Antibiograms were performed for all isolates. Only 73 isolates out of 166 were oxacillin resistant. The percentage of resistant isolates to erythromycin, rifampicin, vancomycin, Ofloxacin, gentamycin, Amoxicillin clavulanic acid, ciprofloxacin, chloramphenicol, trimethoprim sulfamethoxazole, teicoplanin and tetracycline were 58, 32.50, 2.4, 45.18, 37.9, 39.7, 23.5, 21.6, 40.3, 0 and 39.1%, respectively. MRSA isolates were subdivided into eight biotypes according to their resistance pattern. Random amplification of polymorphic DNA (RAPD) and repetitive sequence DNA (REP) were performed on samples representing each biotype. Key words: Methicillin-resistant Staphylococcus aureus (MRSA), nuclease, mecA, 16S rRNA, random amplified polymorphic DNA (RAPD), polymerase chain reaction (PCR).

Aim of study: Otitis media (OM) is highly prevalent worldwide and is the main cause of hearing loss in developing countries. Resistant microorganisms and biofilm formation have become a major health and economic problem. The aim of this study was to establish an eco-friendly method for synthesis of silver nanoparticles using aqueous garlic extract and sunlight. Moreover, detect its potential to inhibit otitis media microbes.Methods: Basically, highly resistant and biofilm forming otitis media pathogenic isolates were determined by Kirby-Bauer disc diffusion method and their biofilm production qualitatively was tested. Then, compared their susceptibility to tetracyclin 30µg (for bacteria) and nystatin 100 µg (for fungi) alone and with different concentrations of green synthesized silver nanoparticles (AgNPs), as well as, their biofilm production. AgNPs were prepared by easy, fast, non-toxic, economically eco-friendly way, using the direct sunlight irradiation on aqueous garlic extract. Their characterization by UV- visible spectral analysis, TEM, DLS, XRD and FTIR were studied. Results: Two multidrug resistant bacterial isolates; Pseudomonas aeruginosa and Bacillus cereus were detected as biofilm producers with black colonies on congo-red agar and three resistant fungal isolates to nystatin; Penicillium chrysogenum, Aspergillus niger and Aspergillus flavus were tested against green synthesized AgNPs. Highly significant (p-value

Egypt ranks first regarding the prevalence of hepatitis C virus (HCV) infection. Many patients have concomitant diseases like kidney disorders requiring hemodialysis, a procedure carrying the hazard of transmitting other hepatitis viruses. The purpose of this study was to investigate for occult hepatitis B virus (HBV), SEN virus (SENV), and torque teno virus (TTV) among chronic HCV patients on maintenance hemodialysis to identify their impacts.A total of 325 hemodialysis patients were enrolled and divided into two groups based on HCV RNA testing results. Blood samples were collected before hemodialysis. Sera were tested for hepatitis B core antibodies (anti-HBc) and hepatitis B surface antibodies (anti-HBs) using ELISA. HBV, SENV, and TTV DNA were detected by PCR. The serum alanine aminotransferase (ALT) level was measured.Anti-HBc and HBV DNA were detected in 73.1% and 50.8% of group 1 versus 36.4% and 22.6% of group 2. The serum ALT level was higher in group 1 than group 2. SENV was detected in 11.5% of group 1 versus 8.2% of group 2. TTV was detected in 29% of group 1 versus 27% of group 2.There is an increased prevalence of occult HBV in our locality among chronic HCV patients undergoing hemodialysis. The existence of SENV and TTV viremia has no clinical impact.

Background & Aims: Hepatitis B is a potentially life-threatening liver infection and it is a major global health problem. Over the past decade, numerous studies have reported that patients with CLD, especially HCV-positive and HBV-positive patients, have decreased 25(OH) D levels. The current study was designed to assess the serum levels of vitamin D [25(OH) D3] in chronic hepatitis B patients, before and during treatment with antiviral therapy. Methods: It was a prospective study in which 80 subjects were enrolled between December 2017 and June 2018. A total of 50 treatment-naïve chronic HBV patients and 30 healthy subjects were recruited. The studied cases received treatment in the form of Lamivudine 100 mg tablet, once daily. Full routine laboratory investigations, HBV DNA measurement by real-time PCR were conducted once before initiation of antiviral treatment and again at least 6 months later. Serum vitamin D level [25(OH)D3 was assessed twice, once before initiation of antiviral treatment and again at least 6 months later. This was done for all the patients enrolled in the study. Results: The studied cases showed a significantly low mean serum Vitamin D level when assessed before treatment (21.6 ± 5.8 ng/ml), compared to the level after 6 ms of treatment (31.1 ± 7.3 ng/ml) which was comparable to that of the control group (33.4 ± 5 ng/ml). Conclusion: The present study highlights the impact of antiviral therapy on vitamin D deficiency in CHB patients, where effective therapy improves vitamin D levels. Meanwhile, it is recommended to study the impact of vitamin D replacement and correction on the disease progression or regression.

Background: Microbial contamination sources in the manufacturing process of some oral pharmaceutical syrups may be a leading cause of diseases in children, elderly and immunosuppressed individuals.Objectives: Traceability of some sources of contamination in manufacturing process of some oral pharmaceutical syrup. Methodology: Analysis of 400 oral syrup samples, raw materials, water and air samples using classical pharmacopeia techniques. Results: In this study, fungal contaminants were more common in pharmaceutical syrup than bacteria and air was an important source of contamination in finished product.

Unfortunately, the affiliation of author "Rania Abdelmonem Khattab" was published incorrectly in the original publication. The correct version of affiliation is updated here.

Clostridium difficile infection (CDI) is a common cause of morbidity and mortality in hospitalized patients worldwide. The major problem facing current treatment is multiple recurrences, prompting the need for alternative therapies. In this study we isolated bacterial species, from Egyptian individuals' stool, with antimicrobial activity against clinical isolates of C. difficile and tried to examine the nature of the produced antimicrobials. In vitro antibacterial activity against C. difficile was initially screened in 123 fecal samples cultures using an agar overlay method. The isolates with antimicrobial activity against C. difficile in addition to Clostridium isolates were identified using partial 16S rDNA gene sequencing analysis. The isolates acting against C. difficile belonged to Lactobacillus, Enterococcus and Clostridium genera. The concentrated cell-free supernatants (CFSs) from these bacterial isolates were examined for antimicrobial activity against C. difficile growth by broth dilution method. 10 x concentrated CFSs of five isolates showed inhibition for C. difficile growth which was significantly different (p 0.001) from control. Lactobacillus agilis T99A and Clostridium butyricum T58A isolates were selected for further evaluation of the produced antimicrobials. The antimicrobial activity of 10x CFSs of the two isolates was stable after enzymatic treatment with proteinase K or heating treatments up to 90 °C or neutralizing pH. The spectrum of activity of the two isolates was evaluated using different gram-positive and gram-negative bacterial species and did not show antimicrobial activity against these species. Our results showed two unconventional bacterial isolates: L. agilis T99A and C. butyricum T58A producing extracellular thermo stable antimicrobial agents against C. difficile clinical isolates.

Introduction: Salmonella typhiVi capsular polysaccharide (ViCPS) is a licensed vaccine against typhoid fever in many countries; in Egypt, the killed whole-cell vaccine is still used. In this study, mice were used as an animal model to evaluate the immune response to ViCPS and other S. typhiantigens such as heat-killed whole cells and outer membrane protein (OMP). Methodology: The three antigens were laboratory prepared, injected into mice groups, and the humoral response was evaluated using the indirect whole-cell enzyme-linked immunosorbent assay (ELISA). The sensitivity of this assay was investigated using in situ or pre-heated whole cells as coating antigens. In addition, the effect of the immunization route for ViCPS was examined. Results: Immunizing doses of heat-killed whole cells as well as ViCPS, 2 and 4 µg given subcutaneously (SC) and 4 µg given intraperitoneally (IP), showed significant immune response compared to controls. However, the responses to these doses were not significantly different from each other. The OMP showed a higher significant response. The sensitivity of indirect whole-cell ELISA was enhanced significantly by in situ heat treatment of the coating antigen rather than the pre-heated coating antigen. Conclusions: The three antigens showed significant immune response. The immune response to OMP was higher. Though heat-killed whole cells and ViCPS are almost similar in immunizing level, ViCPS is recommended. The SC route was more immunizing than the IP one. Furthermore, the sensitivity of the indirect whole-cell ELISA technique could be enhanced by in situ heat inactivation of the coating cells.

The objective of this work was to isolate bacteria from Red Sea invertebrates, determine their antimicrobial activity, and screen for the biosynthetic gene clusters [polyketides (PKs) and nonribosomal peptides (NRPs)] which could be involved in the production of bioactive secondary metabolites. Eleven different samples of marine invertebrates’ were collected from Egypt’s Red Sea (El-Tor-Sharm El-Sheikh and Hurghada) by scuba diving, and a total 80 isolates of the associated microorganisms were obtained from the cultivation on six different cultural medium. Seven isolates of them showed an antimicrobial activity against five pathogenic reference strains, while the most active antimicrobial agent was isolate number HFF-8 which was 99% identical to Bacillus amyloliquefaciens. HFF-8’s extract showed positive results against Gram negative bacteria, Gram positive bacteria and yeast. Moreover, the isolates gave positive bands when screened for the presence of PK synthase (PKS) I and II and NRP synthetase (NRPS) I and II biosynthetic genes, those biosynthetic fragments when cloned and sequenced were primitively predicted as biosynthetic fragments for kirromycin and leinamycin production by NaPDoS program with 56 and 55%, respectively. The Red Sea can provide a sustainable solution to combat bacterial resistance. The contribution of this work is that B. amyloliquefaciens was isolated from Heteroxenia fuscescens, Red Sea, Egypt. Moreover, the bacterial extract showed a broad spectrum with a potent antimicrobial activity.

AIM To determine the possibility of the development of dry eye disease (DED) as a result of persistent infection with Chlamydia trachomatis and Ureaplasma urealyticum in the conjunctiva of patients. METHODS This study was conducted on 58 patients of age range 20-50y, diagnosed with DED confirmed by Schirmer I test and tear breakup time. The non-dry eye control group included 27 subjects of the same age. Ocular specimens were collected as conjunctival scrapings and swabs divided into three groups: the first used for bacterial culture, the second and third taken to detect Chlamydia trachomatis and Ureaplasma urealyticum by direct fluorescent antibody (DFA) assay and polymerase chain reaction (PCR) method. RESULTS Chlamydia trachomatis was detected in 65.5% and 76% of DED patients by DFA and PCR methods respectively. Ureaplasma urealyticum was found in 44.8% of DED infected patients using the PCR method. Both organisms were identified in only 37.9% of DED patients found to be infected. Control subjects had a 22% detection rate of Chlamydia trachomatis by DFA assay versus a 7% detection rate by PCR; while Ureaplasma urealyticum was detected in 3.7% of the controls by PCR method. The conjunctival culture revealed that gram positive microorganisms represented 75% of isolates with coagulase negative Staphylococci the most common (50%) followed by Staphylococcus aureus (20%), whereas gram negative microorganisms occurred in 25% of cases, isolating Moraxella spp. as the most frequent organism. CONCLUSION Our results tend to point out that Chlamydia trachomatis and Ureaplasma urealyticum were detected in a moderate percentage of patients with DED, and could be a fair possibility for its development. PCR is more reliable in detecting Chlamydia trachomatis than DFA technique. The presence of isolated conjunctival bacterial microflora can be of some potential value.

Aim To determine the association between chlamydial conjunctivitis and genital infection by Chlamydia trachomatis, Mycoplasma genitalium and Candida albicans, in addition to the possible relationship between cultured bacterial pathogens and oculogenital chlamydial infection. Methods This study was performed on 100 (50 symptomatic and 50 asymptomatic) women attending the Gynecological and Obstetric outpatient clinic of Alzahra hospital, Alazhar University. Simultaneously a conjunctival swab was taken from these patients. Polymerase chain reaction (PCR) was done on DNA extracted from both vaginal and conjunctival swab samples. Culture for both vaginal and conjunctival swabs was also done. Results Candida albicans was the predominant organism isolated by culture in 20% and 40% of conjunctival and vaginal swabs respectively. By the PCR method, ocular Chlamydia trachomatis was present in 60% of symptomatic women, while genital Chlamydia trachomatis infection was present in 30% of symptomatic women. The results of this method also indicated that 25/50 (50%) vaginal swabs were positive with PCR for Candida albicans versus 15/50 (30%) were PCR positive in conjunctival swab. Mycoplasma genitalium was present in only 10% of vaginal swabs. Concomitant oculogenital PCR positive results for Chlamydia trachomatis and Candida albicans were 30% and 28% respectively. Conclusion Ocular Chlamydia trachomatis was associated with genital Chlamydia trachomatis in a high percentage of women followed by Candida albicans. Cultured bacterial organisms do not play a role in enhancement of Chlamydia trachomatis infection.

There are eight reported cases in the literature of osteosarcomas secreting β-hCG. Our primary aim was to investigate the rate of β-hCG expression in osteosarcoma and attempt to understand the characteristics of osteosarcomas that secrete β-hCG. We reviewed 37 histopathology slides (14 biopsies and 23 surgical specimens) from 32 patients with osteosarcoma. The slides were retrospectively stained for β-hCG expression. Patient and tumour characteristics, including age, gender, tumour location, subtype, proportion of necrosis, presence of metastases and recurrence were recorded. A total of five of the 32 tumours were found to be positive for β-hCG expression (one strongly and four weakly). This incidence of this expression was found in tumours with poor histological response to neoadjuvant chemotherapy. The use of β-hCG expression as a diagnostic, prognostic or follow-up marker is questionable and needs further investigation with a larger sample size.

To determine the possibility of the development of dry eye disease (DED) as a result of persistent infection withThis study was conducted on 58 patients of age range 20-50y, diagnosed with DED confirmed by Schirmer I test and tear breakup time. The non-dry eye control group included 27 subjects of the same age. Ocular specimens were collected as conjunctival scrapings and swabs divided into three groups: the first used for bacterial culture, the second and third taken to detectOur results tend to point out that