Hebatallah Ahmed Nasser

(1) Background: Hyaluronic acid (HA) is a polyanionic mucopolysaccharide extensively used in biomedical and cosmetic industries due to its unique rheological properties. Recombinant HA production using other microbial platforms has received increasing interest to avoid potential toxin contamination associated with its production by streptococcal fermentation. In this study, the Gram-negative strains Escherichia coli (pLysY/Iq), E. coli Rosetta2, E. coli Rosetta (DE3) pLysS, E. coli Rosetta2 (DE3), E. coli Rosetta gammiB(DE3)pLysS, and the Gram-positive Bacillus megaterium (MS941) were investigated as new platforms for the heterologous production of HA. (2) Results: The HA biosynthesis gene hasA, cloned from Streptococcus equi subsp. zoopedemicus, was ligated into plasmid pMM1522 (MoBiTec), resulting in pMM1522 hasA, which was introduced into E. coli Rosetta-2(DE3) and B. megaterium (MS941). The initial HA titer by the two hosts in the LB medium was 5 mg/L and 50 mg/L, respectively. Streptococcal hasABC and hasABCDE genes were ligated into plasmid pP_T7 (MoBiTec) and different E. coli host strains were then transformed with the resulting plasmids pP_T7hasABC and pP_T7hasABCDE. For E. coli Rosetta-gamiB(DE3)pLysS transformed with pP_T7hasABC, HA production was 500 ± 11.4 mg/L in terrific broth (TB) medium. Productivity was slightly higher (585 ± 2.9 mg/L) when the same host was transformed with pP_T7 carrying the entire HA operon. We also transformed B. megaterium (MS941) protoplasts carrying T7-RNAP with pP_T7hasABC and pP_T7hasABCDE. In comparison, the former plasmid resulted in HA titers of 2116.7 ± 44 and 1988.3 ± 19.6 mg/L in LB media supplemented with 5% sucrose and A5 medium + MOPSO, respectively; the latter plasmid boosted the titer final concentration further to reach 2476.7 ± 14.5 mg/L and 2350 ± 28.8 mg/L in the two media, respectively. The molecular mass of representative HA samples ranged from 10⁵ − 10⁶ Daltons (Da), and the polydispersity index (PDI) was E. coli Rosetta-gamiB(DE3)pLysS, while no HA capsules were produced by B. megaterium. (3) Conclusions: Our results suggested that Gram-positive bacteria are probably superior host strains for recombinant HA production over their Gram-negative counters. The titers and the molecular weight (MW) of HA produced by B. megaterium were significantly higher than those obtained by different E. coli host strains used in this study.

The emergence of AmpC (pAmpC) β-lactamases conferring resistance to the third-generation cephalosporins has become a major clinical concern worldwide. In this study, we aimed to evaluate the expression of AmpC β-lactamase encoding gene among the pathogenic Gram-positive and Gram-negative resistant bacteria screened from clinical samples of Egyptian patients enrolled into El-Qasr El-Ainy Tertiary Hospital in Cairo, Egypt. A total of 153 bacterial isolates of the species Pseudomonas aeruginosa, Klebsiella pneumoniae, and Enterococcus faecium were isolated from patients diagnosed with urinary tract infection (UTI), respiratory tract infection (RTI), and wound infections. The total number of E. faecium isolates was 53, comprising 29 urine isolates, 5 sputum isolates, and 19 wound swab isolates, whereas the total number of P. aeruginosa isolates was 49, comprising 27 urine isolates, 7 sputum isolates, and 15 wound swab isolates, and that of the K. pneumoniae isolates was 51, comprising 20 urine isolates, 25 sputum isolates, and 6 wound swab isolates. Our results indicated that there is no significant difference in the expression of AmpC β-lactamase gene among the tested bacterial species with respect to the type of infection and/or clinical specimen. However, the expression patterns of AmpC β-lactamase gene markedly differed according to the antibacterial resistance characteristics of the tested isolates.

Background and Aim: Mastitis has been identified as the most prevalent and economically imperative disease among dairy animals. Thus, understanding its common bacterial pathogens and risk factors is necessary to improve udder health at herd, region, or country level. However, scientific research on caprine mastitis, especially on Beetal breed, has remained to be insufficient in Pakistan. Therefore, this study aimed to evaluate the epidemiology and antibiogram assay of common mastitis-causing bacterial agents, that is, Staphylococcus, Streptococcus, and Escherichia coli, in dairy goats. Materials and Methods: In total, 500 Beetal goats, irrespective of age and those that were not treated with any kind of antimicrobial agents during the past 120 h, were screened using California Mastitis Test in Pattoki, Kasur District, whereas epidemiological factors were recorded. The milk samples of mastitic goats were then collected and processed using standard methods. Each sample was primarily cultured on nutrient agar. Using a specific medium, each bacterial colony was separated using several streak methods. Six antibiotic disks belonging to different antibiotic groups were used for antibiogram profiling of bacterial isolates. Chi-square test was used to assess the association of baseline characteristics and mastitis occurrence. Meanwhile, multivariable logistic regression (p<0.001) was utilized to determine the risk factors associated with positive and negative dichotomous outcome of mastitis. Results: The results revealed that the overall prevalence of goat mastitis was 309 (61.8%), in which 260 (52%) and 49 (9.8%) cases were positive for subclinical mastitis (SCM) and clinical mastitis (CM), respectively. Streptococcus and E. coli were found to be the predominant isolates causing SCM and CM, respectively (p<0.001). It was observed that amoxicillin+clavulanic acid was highly sensitive to isolates of Staphylococcus and Streptococcus and ceftiofur sodium to isolates of Streptococcus and E. coli, while enrofloxacin was found to be sensitive to isolates of Streptococcus and E. coli. Risk factors such as herd structure, deworming, vaccination, presence of ticks, use of teat dip and mineral supplements, feeding type, age, parity, housing, blood in the milk, milk leakage, milk taste, and milk yield were found to have the strongest association with mastitis occurrence, while ease of milking has moderate association. Conclusion: In the area examined, cases of SCM were found to be higher compared with that of CM, and ceftiofur sodium has been identified as the preferred treatment in both clinical and subclinical forms of caprine mastitis in Beetal goats. Risk factors for mastitis that was identified in this study can form the basis for the creation of an udder health control program specific for dairy goats. We hope our findings could raise awareness of the risk factors and treatment approaches for common mastitis-causing bacterial agents.

BACKGROUND: Self-medication is a practice with major global implications, especially with antibiotics intake. Self-medication among future health-care professionals could affect their way in prescribing medication in the future. AIM: This study was conducted to estimate the magnitude and the determinants of antibiotics self-medication practices and to describe the pattern of antibiotics abuse among undergraduate university students. METHODS: A cross-section study was conducted among 563 medical students from public and private universities in Cairo, Egypt, using a questionnaire. RESULTS: About 77.7% of the students used antibiotics without prescriptions with no statistical differences by age, sex, residence, or type of universities. More than half of the students (51.7%) do not know the effect of antibiotics abuse on microbial resistance. Most self-treated antibiotics were used to manage gastroenteritis symptoms (70%), respiratory symptoms (63%), and dental infections (36%), other causes such as headache or prophylactic reasons (21%). About 91.7% of the self-medicated students reported access to antibiotics from the pharmacy without a prescription and 71% of them mentioned discontinuation of a course of antibiotics at least once during the last year. About 81% of the students who do not know the effect of antibiotic abuse are self-medicated versus 75% of their counterpart who know and this difference is statistically significant. The multivariate analysis identified the residence as an independent predictor of their knowledge (area of residence = 1.6, 95% confidence interval [1.1–2.3]). CONCLUSION: The prevalence of self-medication with antibiotics among university students in Cairo is high. Our findings highlight the urgent need for tailored interventions to control this practice.

Mesoporous silica nanoparticles (MSNs) have gained wide attention for their role in biomedicine and as drug delivery vehicles. Their structural tunability, high surface area, and easy functionalization impart significant advantages over conventional materials. In this Review, recent advances in the synthesis, drug delivery, and therapeutic roles of MSNs in the treatment of various neurodegenerative and neuroinflammatory diseases are presented. The intention is to understand how MSN formulations that are capable of encapsulating drug molecules can enhance drug delivery by overcoming the blood–brain barrier (BBB) mediated by specific transport processes. The composition and characteristics of the BBB, and how alterations are observed in neurodegenerative diseases including Alzheimer's, epilepsy, and intracerebral hemorrhage are reviewed. Finally, the factors affecting efficient delivery of MSNs into the brain are summarized, and their most promising functional outcomes are discussed.

Enzymes are powerful versatile biocatalysts, however, industrial application of enzymes is usually hampered by their susceptibility. Bio-inspired Eudragit-α-amylase conjugate (E-AC) was proposed as a biocatalyst for various pharmaceutical and industrial applications. In this study, α -Amylase (E.C. 3.2.1.1) was immobilized by covalent conjugation to Eudragit L-100 under mild conditions. The effect of polymer, carbodiimide and enzyme concentrations on optimization of (E-AC) was investigated. In addition, characterization of the free α -Amylase and E-AC with regard to pH, temperature, kinetic parameters, reusability and operational and storage conditions was carried out. Results showed a shift of the optimum pH of E-AC towards the alkaline side whereas, E-AC exhibited higher thermal stability at all tested temperatures. The kinetic parameters, K_m values were 2.87 mg/ml and 3.15 mg/ml and V_max values were 8.35 mg/ml/min and 8.98 mg/ml/min for free and E-AC, respectively. E-AC retained 85% of the initial activity after five consecutive amylolytic cycles, thus emphasizing its powerful potentials. Operational storage and thermal stability were highly improved as well for E-AC conjugate with an 11.6 stabilization factor in comparison to the free α-amylase. In this study, Eudragit L-100 polymer was successfully used as smart immobilization support to create a reversibly soluble-insoluble enzyme biocatalyst to enforce and extend biotechnological applications of α-amylase in the pharmaceutical industry.

Summary: Background: Recent studies suggest asthma prevalence in polycystic ovary syndrome (PCOS) patients. This is the first study to explore asthma prevalence among Egyptian PCOS patients. It highlighted common findings in PCOS and asthma. It investigated whether these findings could serve as potential predictors of asthma.Methods: A hundred PCOS patients, sixty asthmatic patients and thirty apparently healthy females of matched age were included. Body mass index (BMI), C-reactive protein (CRP), IL-6, IgE, 25 (OH) vitamin D, testosterone and lipid profile were measured.Results: Both PCOS and asthmatics had significantly higher BMI, Total cholesterol (TC), LDL-C, IgE, CRP and IL-6 (P2and 117.6 nmol/L respectively could differentiate between asthmatic and non-asthmatic PCOS with sensitivity 63.8 % and specificity 62% for BMI, and sensitivity and specificity of 66% for CRP.Conclusions: This study shows that BMI and CRP are predictors of asthma development in Egyptian PCOS.