Determination of some antiemetic drugs through its native fluorescence or fluorescence quenching of cerrous ammonium sulphate.

The manuscript describes two fluorimetric methods for the determination of some antiemetic drugs namely granisetron HCl, ondansetron HCl and tropisetron HCl, used in the management of nausea and vomiting induced by cytotoxic chemotherapy and radiotherapy. Granisetron HCl solution exhibits a native fluorescence, which can be applied for its determination at 365 nm upon excitation at 305 nm. The method was applied for the determination of granisetron HCl in drug substance, drug product as well as in presence of its acid induced degradation products. The quantum yield was calculated. The second proposed method is based on measuring the quenching effect induced by ondansetron HCl or tropisetron HCl on the fluorescence intensity of cerrous ammonium sulphate at λem 348 nm upon excitation at 250 nm in acidic medium. The analysis of quenching data showed that quenching of cerrous ammonium sulphate induced by ondansetron HCl or tropisetron HCl is mainly through dynamic quenching. Various variables affecting fluorescence response were studied and optimized. The obtained results were found to be statistically agreed with those obtained from the official or reported ones. Moreover, the validity of the methods was assessed according to ICH guidelines.

Could aflatoxin B1 production by Aspergillus flavus affect the severity of keratitis: an experience in two tertiary health care centers, Egypt.

To investigate the expression of AFB1 gene in isolates obtained from corneal scrapping samples from keratitis patients and to correlate the quantity of AFB1 to the severity of keratitis. An observational study was undertaken in Medical Microbiology and Immunology department, Mansoura University, Egypt, over corneal scrapping samples that were cultured aiming to isolate fungal causes of infective keratitis followed by AFB1 gene detection in Aspergillus flavus isolates by nested PCR then quantitation of the toxin by TLC. Out of 843 corneal scrapping samples collected from patients with infective keratitis, positive fungal growth was identified in 277 cases (32.9%). A. flavus was the commonest fungal agent isolated in 93 cases (33.6%). The AFB1 toxin-encoding gene was detected in 63.4% of A. flavus isolates. There was a positive correlation between the quantity of produced AFB1 toxin and the degree of severity of keratitis (P value < 0.0001*). Aspergillus flavus was the most common cause of fungal keratitis, with the AFB1 toxin-encoding gene detected in more than half of the isolates. A significant correlation between the degree of severity of keratitis and the quantity of produced AFB1 toxin was detected. Therefore, exploring presence or absence of AFB1 toxin is important for the clinicians in their diagnostic assessment and selection of proper treatment choices.

Validated Liquid Chromatographic Method for the Determination of (Canagliflozin, Dapagliflozin or Empagliflozin) and Metformin in the Presence of (1-Cyanoguanidine).

A simple and accurate liquid chromatographic method has been developed and validated for the determination of either canagliflozin, dapagliflozin propandiol monohydrate or empagliflozin and metformin in presence of metformin major degradation product;1-cyanoguanidine. The Liquid Chromatographic (LC) method was based on isocratic elution on Prontosil (Lichrosorb 100-5-NH2) column using a mobile phase consisting of NaH2PO4 buffer (10 mM, pH 2.8):acetonitrile (18.5:81.5, v/v), at a flow rate of 2 mL/min-1. Quantitation was achieved with UV detection at 225 nm. The validation of the method was assessed according to International Conference on Harmonization (ICH) guidelines. Linearity, accuracy and precision were satisfactory over the concentration ranges of 12.5-100, 3.75-30, 0.3075-2.46, and 0.3125-2.5 µg/mL for metformin HCl, canagliflozin, dapagliflozin propandiol monohydrate and empagliflozin, respectively. Limits of detection and quantitation were found to be 0.068, 0.135, 0.077 and 0.069 µg/mL and 0.206, 0.410, 0.233 and 0.210 µg/mL for metformin HCl, canagliflozin, dapagliflozin propandiol monohydrate and empagliflozin, respectively. The developed method is suitable for the quality control and routine analysis of the cited drugs separately or in combinations.

Serum CD64 and ascitic fluid calprotectin and microRNA-155 as potential biomarkers of spontaneous bacterial peritonitis.

AIM: Patients with ascites are at a higher risk for associated of on top bacterial infections with subsequent life-threatening complications. We aimed to evaluate CD64, calprotectin, and microRNA-155 (miR-155) levels as diagnostic markers of spontaneous bacterial peritonitis (SBP) and the effect of using more than one use on the same spot over their diagnostic efficiency. PATIENTS AND METHODS: An observational comparative study included 103 patients with ascites admitted to the Tropical Medicine Department, Mansoura University Hospital, Egypt, divided into two groups: case group (64 patients) with ascites with SBP and control group (39 patients) with decompensated cirrhotic non-SBP ascites. Twenty milliliters of ascetic fluid was obtained from all participants for bacterial culture, and assessment of calprotectin and miR-155, in addition to 2 ml blood for the CD64 marker expression assay by a flowcytometer. RESULTS: The sensitivity and specificity of CD64 expression assay were 95.3 and 92.3%, respectively, area under the curve (AUC)=0.93, whereas those of ascetic fluid calprotectin and miR-155 were 87.5 and 82.1%, AUC=0.90 and 95.3 and 97.4%, with AUC of 0.95. Combined blood CD64 and ascetic fluid calprotectin had a diagnostic accuracy of 0.988 for blood CD64 and ascetic fluid miR-155, AUC=0.991, and that for ascetic fluid calprotectin and miR-155 was 0.988. On using the three studied markers together, the diagnostic accuracy was the best recorded, AUC=0.994. P values were less than 0.001. CONCLUSION: CD64, calprotectin, and miR-155 were good diagnostic markers of SBP and on using this combination, greater efficiency in diagnosis was achieved.

Development and Validation of Spectrophotometric Methods for the Determination of Canagliflozin or Gliclazide and Metformin in the Presence of Metformin Impurity (1-Cyanoguanidine).

Background: Quantitative multicomponent analysis is considered an analytical goal to save time and cost in analysis. Objective: This work aimed to provide sensitive and selective [successive ratio subtraction coupled with constant multiplication (SRS-CM) and chemometric] methods for the determination of coformulated antidiabetic drugs, namely canagliflozine and metformin or gliclazide and metformin in presence of metformin degradation product, 1-cyanoguanidine. Methods: SRS-CM method was developed for the determination of canagliflozin and metformin at 292 and 237 nm, respectively, using 14 µg/mL canagliflozin as a divisor in the first step to cancel the spectrum of canagliflozin. Then, 18 µg/mL metformin was used as a divisor in the second step to cancel the spectrum of metformin. Finally, we obtained the spectrum of cyanoguanidine. Chemometric method was applied for the determination of the gliclazide and metformin mixture in a 225-235 nm range. Sample enrichment technique was used to increase the concentration of canagliflozin or gliclazide to allow its simultaneous determination with metformin without prior separation. Results: Validation parameters according to the International Conference on Harmonization guidelines were satisfactory over the concentration ranges of 5 to 16 µg/mL for canagliflozin and metformin as well as 2.5 to 12.5 and 6 to 24 µg/mL for gliclazide and metformin, respectively. Conclusions: The method provides sufficient selectivity and accuracy to be applied for routine analysis and quality control in laboratories for the cited drugs. Highlights: This work shows two examples to how to select a suitable UV spectrophotometric method to overcome the spectral overlap.

High-risk and low-risk human papilloma virus in association to spontaneous preterm labor: a case-control study in a tertiary center, Egypt.

INTRODUCTION: This study aimed to detect the correlation between human papillomavirus (HPV) and spontaneous preterm labor in Egyptian women and its association to the human papilloma viral load and MPP2 gene expression. MATERIAL AND METHODS: We performed an observational comparative case-control study in Department of Obstetric and Gynecology, Mansoura University Hospitals over women presented with spontaneous preterm labor, besides females admitted for giving birth at full term to detect conserved sequence in HPV-L1 gene (GP5/GP6) followed by genotype detection of high- and low-risk HPVs with quantification of the viral load and the MMP2 gene expression using real-time polymerase chain reaction (PCR). RESULTS: The prevalence of HPV was 18.1% in preterm females, but only 4% in full-term women (p value = 0.019*). Twenty percent were PCR positive for HPV 16 and 40% for HPV 18 whereas none of the control was positive for any of the studied high-risk genotypes. Thirty percent were PCR positive for HPV 6 and 10% were positive for HPV 11. MMP2 gene expression was significantly higher in preterm than full term. Human papilloma viral load was found to be positively correlated to the rate of MMP2 expression and the gestational age was significantly related to the viral load and the rate of expression of MMP2 gene. CONCLUSION: Human pabilloma virus especially high-risk genotypes was correlated to spontaneous preterm labor in Egyptian females through increasing early expression of MMP2 gene. The time of occurrence of preterm labor was affected by the viral load and so the rate of expression of MMP2 gene.

Correlation of mupirocin resistance with biofilm production in methicillin-resistant Staphylococcus aureus from surgical site infections in a tertiary centre, Egypt.

The aim of this study was to detect mupirocin-resistant isolates from pus/wound swabs taken postoperatively in a tertiary centre in Egypt and to determine their ability to form biofilm in order to establish its correlation with mupirocin resistance. This was a prospective study including 513pus/wound swabs from patients suffering from postoperative surgical site infections over the period July 2013-January 2015. Samples were cultured and isolates were identified by coagulase activity, DNase test, mannitol fermentation by mannitol salt agar followed by API Staph 32. Oxacillin agar screen test, agar dilution test for mupirocin, and mupA gene detection by PCR were performed for all methicillin-resistant Staphylococcus aureus (MRSA) isolates. Biofilm detection was carried out by the microtitre plate and Congo red agar methods. Of the 161 S. aureus isolates identified, 73 (45.3%) were MRSA, among which 82.2% were mupirocin-susceptible and 17.8% were mupirocin-resistant. Among the resistant isolates, 38.5% showed low-level resistance and 61.5% were high-level mupirocin-resistant. The mupA gene was detected in 75.0% of high-level mupirocin-resistant strains and in none of the low-level mupirocin-resistant strains. Among the mupirocin-susceptible isolates, 95.0% were biofilm-producers and 5.0% did not produce biofilm. All mupirocin-resistant isolates produced biofilm. Moreover, 15.3% of high-level mupirocin-resistant strains were negative for the mupA gene but showed evidence of biofilm formation. In conclusion, biofilm formation may be suggested to play a role in mupirocin resistance besides the presence of a genetic element encoding abnormal isoleucyl-tRNA synthetase, however further studies are needed to confirm these findings.

UreA and cagA genes of Helicobacter pylori in Egyptian patients with laryngeal squamous cell carcinoma and benign laryngeal polyps: a cohort study.

This work aims to estimate the prevalence of Helicobacter pylori ureA gene and evaluate cagA gene-positive strains in both patients of laryngeal squamous cell carcinoma (LSCC) and those with benign laryngeal polyps. This study included 49 patients confirmed pathologically to have LSCC and 15 patients with benign laryngeal polyps over a period from June 2013 to March 2015. Samples of laryngeal tissue were collected during direct laryngoscope under general anesthesia to be pathologically evaluated followed by analysis for H. pylori detection. Each laryngeal tissue sample was divided into three parts; one for bacteriological examination, the second for pathological examination and the third for PCR to detect both ureA and cagA genes. Out of 49 LSCC samples, 31 (64.6 %) was positive for ureA by PCR. Out of them, 29 samples (93.5 %) were cagA positive. Only three cases (20 %) of the benign laryngeal polyp were ureA positive by PCR and one of them was cagA positive by PCR. By the bacteriological culture, only eight samples (25.8 %) gave growth. All of them were ureA positive and only seven of them were cagA positive. There was a significant association between presence of H. pylori and LSCC as compared to benign laryngeal polyp which may contribute in the pathogenesis of laryngeal carcinoma. These results should be confirmed by further studies over larger number of cases.