Colistin-resistance genes in Escherichia coli isolated from patients with urinary tract infections.

BACKGROUND: The incidence of antimicrobial resistance is alarmingly high because it occurs in humans, environment, and animal sectors from a "One Health" viewpoint. The emergence of plasmid-carried mobile colistin-resistance (MCR) genes limits the efficacy of colistin, which is the last-line treatment for multidrug resistance (MDR) against gram-negative infections. OBJECTIVES: The current study aimed to investigate emergence of colistin-resistance (MCR 1-5) genes in E. coli isolated from patients with urinary tract infections (UTIs) in Jordan. METHODS: E. coli (n = 132) were collected from urine specimens. The E. coli isolated from human UTI patients were examined the resistance to colistin based on the presence of MCR (1-5). All isolates were tested against 20 antimicrobials using the standard disk diffusion method. The broth microdilution technique was used to analyze colistin resistance. In addition, the MCR (1-5) genes were detected using multiplex PCR. RESULTS: Out of the 132 isolates, 1 isolate was colistin-resistant, having a minimum inhibitory concentration of 8 µg/mL and possessing MCR-1. All the E. coli isolates showed high resistance to penicillin (100%), amoxicillin (79.55%), cephalexin (75.76%), nalidixic acid (62.88%), tetracycline (58.33%), or cefepime (53.79). CONCLUSION: To our knowledge, this is the first report on the presence of plasmid-coded MCR-1 in E. coli from a patient with UTIs in Jordan. This is a problematic finding because colistin is the last-line drug for the treatment of infections caused by MDR gram-negative bacteria. There is a crucial need to robustly utilize antibiotics to control and prevent the emergence and prevalence of colistin-resistance genes.

Association of Genetic polymorphisms of EDN1 gene and Endothelin-1 level in patients with type 2 diabetes mellitus in the Jordanian population.

Endothelin-1 (ET-1) is one of the most potent vasoconstrictors, encoded by the endothelin-1 (EDN1) gene. It has been shown to play an important role in different diseases including Diabetes Mellitus (DM). Various single nucleotide polymorphisms (SNPs) in the EDN1 gene are related to microvascular complications of type 2 diabetes mellitus (T2DM) such as retinopathy, neuropathy and nephropathy. This study aims to determine the association between two selected EDN1 gene polymorphisms (rs2071942 G > A, rs5370 G > T) and T2DM in the Jordanian population, also to measure the level of ET-1 in T2DM. The samples were collected from the National Center of Diabetes, Endocrinology, and Genetics- Amman, Jordan, including 97 patients with T2DM and 80 healthy individuals. PCR-RFLP was used for SNPs genotyping. ET-1 level was determined using IQELISA kits. The univariate analysis for both SNPs didn't show statistically significant differences in the genotype or allele frequencies among T2DM cases as well as in controls. The same results were obtained regarding ET-1 concentration. The subgroup analysis by sex showed that the genotype and allelic frequencies of rs5370, rs2071942 G/A polymorphisms were not significantly different in males and females. Multivariate Analysis adjusted for various confounders didn't express statistical significance difference for occurrences of both SNPs. However, height and gender showed to be significant risk factors for occurrences of heterozygote alleles in both SNPs. On the other hand, the duration of diabetes has appeared to be related to the recessive allele in rs5370.

DRUG MISUSE AND SELF-MEDICATION AMONG PHARMACY STUDENTS IN JORDAN.

OBJECTIVE: Aim: To estimate risks and prevalence of self-medication and potential abuse risk among pharmacy students in Jordanian Universities. PATIENTS AND METHODS: Materials and Methods: A cross-sectional study design was conducted with 450 students, selected using multistage sampling methods, from seven different universities. Data was collected by self-administrated questionnaires covering demographic and academic information, health-related information, use of self-medication, and pattern of self-medication among pharmacy students. RESULTS: Results: Out of 394 students who answer the questions, 76.9% reported that they had usually treated themselves in case of simple cases without physician or pharmacist consultation. Most commonly used drugs among the surveyed students were Paracetamol 60%, multivitamins supplement 74.25%, and herbal products 37.2%, combination of NSAIDs and Paracetamol 20.6%, and laxatives 19.4%. Cold and flu 25.5%, headache 22.3%, abdominal pain 7.9%, gastric pain 7.9%, cold and flu, headache, abdominal pain, and gastric pain 14.9% were the main conditions which contribute to self-medication practice. It was also found that Pharmacy students were over-confident with the type of cases they could treat without referral to a specialist physician, despite knowing that some of the symptoms may be due to serious health problems. Misuse of analgesics and laxatives was clear, and there was a weakness in knowledge of the indications for the use of the most common drug. CONCLUSION: Conclusions: The prevalence of self-medication among pharmacy students in Jordan is high, and medical teaching institutions need to educate students about the proper use of medicines. Strict legislation and more education on self-medication are necessary for effective use of medicines.

Association of KDR rs1870377 genotype with clopidogrel resistance in patients with post percutaneous coronary intervention.

BACKGROUND: Clopidogrel is an antiplatelet therapy that is widely used in pre and post percutaneous (PCI) coronary intervention procedures to prevent platelet aggregation and stent restenosis. However, there is a wide inter-individual variation in clopidogrel response and some patients showed resistance against the activity of Clopidogrel. Kinase insert domain receptor (KDR) gene is responsible for the transcription of vascular endothelial growth factor receptor 2 (VEGFR2) that plays a major role in the cardiovascular diseases (CVDs) and platelet aggregation. The aim of this study was to find out the association of KDR rs1870377 genotype with clopidogrel resistance (CR) in CVD patients, of Iraqi Arabic origin, hospitalized for elective PCI. MATERIALS AND METHODS: This study was a case-control study with a total of 324 PCI patients. Those patients were classified into 213 patients with non-clopidogrel resistant and 111 patients with CR, depending on the analysis of platelet activity phenotype after clopidogrel administration. KDR rs1870377 was genotyped for all patients using polymerase chain reaction-restriction fragment length polymorphism technique and confirmed by DNA Sänger sequencing through applying Biosystems Model (ABI3730x1). RESULTS: KDR rs1870377 SNP is strongly associated (Chi-sqaure, p vale <0.05) with CR under dominant, co-dominant and recessive models. Additionally, A allele in the rs1870377 SNP may have an impact on the serum levels of VEGFR2 and low density lipoprotein. CONCLUSIONS: KDR rs1870377 SNP is a potential genetic biomarker of CR among CVD patients of Iraqi Arabic origin. Further clinical studies, with larger sample, are required to confirm the findings of this study.

Antimutagenic activities of two suspected anticarcinogenic bifunctional organoiron seleno-terephthalate derivatives.

Two newly bifunctional organoiron seleno-terephthalate derivatives (S1 and S2) were synthesized as potential anticarcinogenic compounds. In a previous study, they were found to have antibacterial and/or antifungal activity, while they did not show any mutagenic action. Such compounds were investigated in the present study for their antimutagenic activity. Sodium azide, hydrogen peroxide, and 4-nitro-o-phenylenediamine, as known mutagens for strains TA100, TA102, and TA98 of Salmonella typhimurium, respectively, were used. Both (S1 and S2) compounds showed a strong antimutagenic action of >98% against sodium azide, >70% against hydrogen peroxide, and >65% activity against 4-nitro-o-phenylenediamine. Bearing in mind the strong correlation between mutagenicity and carcinogenicity, the above compounds can be considered as potentially promising anticarcinogens. Therefore, the present results are very encouraging to investigate the above compounds for other biological activities, including their evaluation as anticarcinogens. A suggested mechanism for the antimutagenicity of the tested compounds is presented.

Genotoxicity of different tert-butylcalix[4]crowns.

The ability of two calix[4]arene derivatives, namely 25,27-p-tert-butylcalix[4]dithiooxabenzocrown (1) and 25,27-p-tert-butylcalix[4]trithiooxabenzocrown (2), to produce chromosomal aberrations in root meristematic cells of Allium cepa and micronuclei (MN) in normochromatic erythrocytes (NCE) of Balb/c mice was investigated. NCE are normal mature red blood cells with a full complement of hemoglobin but lack ribosomes. In the first test, the root tips were treated with a series of concentrations of the two test chemicals ranging from 10(-7) to 10(-4) M for 24 or 48 h. Both compounds caused concentration-dependent increases in the percentage of aberrant cells and reductions in the mitotic index. These effects depended, to some extent, on the duration of the treatment. The most conspicuous chromosomal abnormalities were c-mitosis, chromosome bridges, chromosome breaks, chromosome lags as well as micronuclei and multinuclei. In the second test, acridine orange fluorescent staining was applied to evaluate the incidence of MN in NCE of mice intraperitoneally injected with varying contents of the two test chemicals (0.02-0.08 mg/mouse). The two chemicals induced dose-dependent MN formation as compared to the negative control. The second compound had more pronounced cytogenetic influence than the first one. Mitomycin C (MMC, 14 mg/kg body weight), employed as positive control, produced more obvious effects on the parameters investigated.

Study of the effects on DNA of two novel nucleoside derivatives synthesized as potential anti-HIV agents.

The pursuit of antiviral active compounds against different classes of viruses, in particular HIV, HBV, and HTLV is an area of important and intense research. In the current study, two novel nucleoside derivatives belonging to a new class of isoxazolidine were successfully synthesized as potential anti-HIV agents by replacement of the furanose ring by a N,O-heterocyclic ring Both compounds were investigated for biological activity, namely, mutagenic and antimutagenic properties. Using Salmonella typhimurium strains TA97, TA100, and TA102, both compounds proved to be nonmutagenic, which may be considered an encouraging result to further elucidate other biological activities. Antimutagenic testing of the synthesized compounds revealed that they are active against the base-pair substitution mutagen sodium azide. However, they did not show any indication as antimutagenic agents against hydrogen peroxide and mitomycin C (oxidative mutagens) or against nitrophenylenediamine (a base-pair substitution and frameshift mutagen). Structure-activity relationship is also discussed. Testing these compounds as antiviral agents is highly recommended.

A notable antimutagenicity of two nonmutagenic novel oxadiazoles in Salmonella mutagenicity assay.

The mutagenic activity of two newly synthesized oxadiazoles: 1,3-bis(5-benzylthio-1,3,4-oxadiazol-2-yl) benzene (M1) and 1,4-bis(5-benzylthio-1,3,4-oxadiazol-2-yl) benzene (M2) was studied in Salmonella typhimurium strains TA97, TA100, TA102 and TA1537 in the presence and absence of S9mix. The antimutagenicity of M1 and M2 against H2O2, sodium azide (SA) and 4-nitro-o-phenylene diamine (NPD) using the tester strains TA102, TA100 and TA97, respectively, was also investigated. The two compounds were found to be nonmutagenic using the four tester strains. However, they showed high mutagenic repression activity against hydrogen peroxide (95% and 97% for M1 and M2, respectively, at a concentration of 335 micrograms/plate). Moderate mutagenic repression against NPD (58% and 55% for M1 and M2, respectively, at a concentration of 167.5 micrograms/plate) and low mutagenic repression against SA (21% and 33% for M1 and M2 respectively, at a concentration of 335 micrograms/plate) was detected. The obtained results are very encouraging to test the above mentioned compounds as anticarcinogens.

In vitro cytogenetic testing of an organoselenium compound and its sulfur analogue in cultured rat bone marrow cells.

BACKGROUND: Selenium (Se) is a non-metal element, occurring in varying degrees in the environment and it has been found to be a component of several enzymes. Different selenium compounds have been associated with carcinogenicity, toxicity, modification of metal toxicity and prevention of cancer. Organoselenium compounds had substantially greater bioavailability and less toxicity than that of inorganic selenium. From a chemical point of view, Se resembles sulfur (S) in many of its properties, thus, Se and S may be considered to be isosteric. The ability of a synthetic organoselenium compound; cyclopenta-dienyldicarbonyl ironselenoterephthalic acid (CSe) and its sulfur analogue (CS) in the range of 10-8 to 10-5 M, to induce sister-chormatid exchanges (SCE) and alter cell division expressed as mitotic index (MI) as well as cell survival has been investigated. METHODS: Rat bone marrow cells were cultured in the presence of CSe and CS in the range of 10-8 to 10-5 M with a total exposure time of 4, 16 or 28 h at 37 degrees C. Fluorescence-plus-Giemsa (FPG) technique was used to visualize chromosomes for SCE analysis and MI determination. Trypan blue exclusion technique was used to determine cell viability. RESULTS: At the three exposure times, cell survival progressively decreased with increasing concentration, but the effect of either chemical was not significant (ANOVA; P < 0.05) as compared to the negative control. Significant reductions in MI were calculated at the highest concentration (10-5 M) when either chemical was applied for 16 or 28 h. Furthermore, the mean SCE increased with longer exposure times and, in general, CSe had slightly greater effect on cell survival and caused higher frequencies of SCE than CS. The exception was the 10-8 M treatment. However, both CSe and CS failed to induce 2-fold SCE as that of the negative control and therefore they are not considered as mutagens. CONCLUSION: Both CSe and CS in the range of 10-8 to 10-5 M could not double the SCE rate of the negative control and therefore not considered as mutagens at these experimental conditions.

Element analysis and biological studies on ten oriental spices using XRF and Ames test.

Ten oriental spices were analyzed for their element composition using X-ray fluorescence (XRF): nutmeg (Myristica fragrans), coriander (Coriandrum sativum), safflower (Carthamus tinctorius), caraway (Carum carvi), Sicilian sumac (Rhus coriaria), aniseed (Anisum vulgare), black pepper (Piper nigrum), cardamom (Elettaria cardamomum), cumin (Cuminum cyminum) and nigella (Nigella sativum). The spices were found to contain the following elements: Mg, Al, Si, P, S, Cl, K, Ca, Ti, Mn, Fe, Cu and Zn, with varying concentrations. Mutagenic studies using Salmonella typhimurium strains TA97a, TA98, TA100, and TA102 showed that the above spices have no base pair substitution mutagenic activity. However, a weak frameshift mutagenicity has been shown by nutmeg and a very weak oxidative mutagenic action has been revealed by cumin.

Synthesis, antibacterial, antifungal and genotoxic activity of bis-1,3,4-oxadiazole derivatives.

In the present investigation, four 1,3,4-bis-oxadiazole derivatives were synthesized as potential antimicrobial agents. The compounds are: 5,5'-dimercapto-bis-[1,3,4-oxadiazol-2-yl]propane (2a), 5,5'-dimercapto-bis-[1,3,4-oxadiazol-2-yl]butane (2b), 5,5'-dimercapto-bis-[1,3,4-oxadiazol-2-yl]octane (2c) and 5,5'-dibenzylthio-bis-[1,3,4-oxadiazol-2-yl]butane (3). The above newly synthesized compounds were investigated for their antibacterial, antifungal and mutagenic activities. The results of the biological activities revealed that the compounds 2a-c exhibited both antibacterial and antifungal activities against S. aureuss and B. subtilis. Compound 2a also showed activity against P. aeureoginosa. All the above compounds and compound 3 exhibited activity against C. albicans. Genotoxic studies showed that compound 2a had a weak base pair substitution mutagenicity but none of them exhibited a frameshift mutagenic action using Ames test.