Molecular diversity of Klebsiella pneumoniae clinical isolates: antimicrobial resistance, virulence, and biofilm formation.

One of the mechanisms responsible for antibiotic resistance in Klebsiella pneumoniae is the enzymes produced by the bacteria; another important mechanism is the ability to form biofilm. In this study, antibiotic resistance, genes associated with virulence, and biofilm-forming properties of K. pneumoniae strains were investigated. A total of 100 K. pneumoniae isolates were obtained from different clinical samples identified by Matrix-Assisted Laser Desorption/Ionization time-of-flight Mass Spectrometry. Antimicrobial susceptibility testing was performed with the Phoenix 100 apparatus. The biofilm forming properties of strains were determined by the microtiter plate method. For molecular analysis, genes encoding the carbapenemase enzyme (blaOXA-48, blaNDM-1, blaIMP, and blaVIM) and biofilm-related genes (treC, luxS, mrkA, and wza) were investigated by polymerase chain reaction (PCR). While 76% of clinical isolates were resistant to three or more antimicrobials, 24% were classified as non-multidrug resistant (non-MDR). When biofilm-forming capacities of clinical isolates were tested, it was determined that the resistant-isolates produced 59.2% strong biofilm, and susceptible-isolates produced 12.5% strong biofilm. According to PCR results, carbapenemase genes were determined as follows: blaOXA-48-70%, blaNDM-49%, and blaKPC-19%, blaOXA-48/blaNDM/blaKPC-12%, blaOXA-48/blaNDM-26%, and blaOXA-48/blaKPC-4%. The biofilm-associated genes in bacterial isolates were determined as follows: luxS-98%, treC-94%, mrkA-88%, and wza-15%. In addition, Hierarchical Clustering Tree and Heatmap analysis revealed an association between isolates that lacks resistance genes and isolates lacks biofilm-formation related genes that were included in MDR or non-MDR classes. As a result, biofilm should be considered in the treatment of MDR infections, and therapy should be planned accordingly. In addition, pursuing the data and genes of antibiotic resistance is significant for combating resistance.

Methamphetamine as the most common concomitant substance used with pregabalin misuse.

BACKGROUND AND AIM: Non-medical use of Pregabalin (PGB) is a growing concern in many countries because of the serious consequences associated with their abuse. Judicial cases within the probation system, multiple drug users, and patients in treatment programs administered PGB at higher doses than suggested, commonly without prescription. For this reason, it is important to analyze PGB by adding it to the routine analysis scale in determining whether PGB is used for medical purposes or abuse. In this study, PGB analyzed (single or multiple substance use, concomitant substances) in urine samples of forensic and clinical cases by liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition to the sociodemographic and clinical characteristics of pregabalin-positive cases, the results were evaluated separately from a clinical and forensic perspective. METHODS: All urine samples which was admitted to Addiction Toxicology Laboratory from 'drug abuse probation system' (forensic cases, n = 640) and from various departments of our hospital (clinical cases, n = 371) between December 2022 and April 2023. Screening analysis were carried out by immunoassay in total 1011 cases. LC-MS/MS method simultaneously analyzed amphetamine, benzoilecgonine, cocaine, codeine, metamphetamine, morphine, 3,4-metilenedioksi-N-metilamfetamin (MDMA), 11-nor-9-karboksi-Δ9-tetrahidrokannabinol and pregabalin in urine samples. PGB was added to the our routine substance screening analysis scale in December 2022 to detect pregabalin use. RESULTS: PGB was detected in 12.3% of probabition cases and 13.2% of clinical cases. The mean age of PGB positive cases was 26.55 ± 7,52 years old, predominantly males (%85,9). Single PGB was detected in 53.2% of forensic cases (n = 42), and 38.7% of clinical cases (n = 19). The most common substance detected concomitantly with PGB was amphetamine type stimulants (ATSs:amphetamine, methamphetamine, ecstasy/MDMA etc.) (22.8% of forensic cases and 46.9% of clinical cases), followed by concomitant cannabis use (24.1% of forensic cases and 26.5% of clinical cases). Concomitant opioid use was rare (1.3% of forensic cases and 4.1% of clinical cases). Detection of PGB was significantly different across months on which the samples were collected (x2 = 82.8, df=4, p < 0.001). CONCLUSION: Inconsistently with previous studies suggesting opioids as the most prevalant substances concominant with PGB, our results showed that stimulants (especially ATSs) were the most prevelant substances concominant with PGB, followed by cannabis. High proportion of PGB detection in probabition cases, frequently as a single substance abuse takes attention. These results suggest that PGB, may be used to avoid legal consequences. It is important for laboratories to be aware that they need to make changes as addition of newly abused substances in their analysis panels, when necessary, as differences between regions and cultures affect substance use patterns.

Pregabalin Qualitative Detection in Turkish Forensic Cases Between 2017 to 2018.

Pregabalin (PGB) has been commonly subjected to diversion, from being a prescription drug to a recreational drug. In this study, pregabalin use (which is one of the substances subject to control in Turkey) and the concomitant use of other substances in cases admitted to the Izmir Forensic Medicine Institute was evaluated. Samples from 15,259 cases were screened, between June 2017 and December 2018, for the presence of PGB, and PGB positive cases were further analyzed. Of all cases screened, PGB was detected in 3.2% (n = 487). The mean age of PGB positive cases was 29.24 ± 10.34 years old (min: 14, max: 84), and 94% of them were male. Cannabis metabolite THC-COOH was the most common substance detected in the blood samples following PGB. Overall, 8 other substances were commonly used along with PGB. These substances were cannabis, morphine, hydromorphone, codeine, hydrocodone, heroin, paracetamol, and naproxen. Finally, we observed similar results in urine analysis. This research provides systematic data for PGB use in forensic cases in Turkey. The study findings indicate that PGB and multiple drug use increased over time, and providers should be particularly careful when prescribing PGB.

Development of analytical method for illegal substances in sweat and comparison of the effectiveness of sweat collection materials.

PURPOSE: Sweat analysis can yield valuable information in forensic investigations, diagnosis and treatment. The purpose of this study was to develop a validated gas chromatography-mass spectrometric method for the detection of illegal substances in sweat after optimizing the method by chemometric approach. This study also investigated the effectiveness of alternative sweat-collecting materials. METHODS: Plackett-Burman screening design was employed to determine the effect of seven process factors on this new method. Then, central composite design (CCD) was used to optimize the method. The method was validated according to the international guidelines. The effectiveness of alternative sweat-collecting materials (cosmetic pads and swabs) was compared with a commercially available collecting device (DrugWipe5A). RESULTS: Sample pH, ultrasonic bath time, and liquid-liquid extraction (LLE) shaking time were determined as the most significantly effective three parameters with the Plackett- Burman screening design. The validation procedure was successfully performed after optimizing this method. The comparison study demonstrated that cosmetic pads, swabs, and DrugWipe5A can be used interchangeably. CONCLUSIONS: Our results suggested that the statistical optimum strategy was an effective tool for the optimization of process parameters. Combined with the sensitivity and selectivity of our method, the analysis of sweat collection materials proved to be a useful tool for physicians and health care professionals.

Toxicological evaluation of alcohol and substance abuse in children and adolescents.

Unhealthy behaviors such as use of alcohol and drug usually begin during adolescence. Izmir is on transit route for illicit substance due to geographical situation. Children and adolescents are the most important threatened group in terms of alcohol and substance abuse. In this study, it was aimed to investigate alcohol and substance use profile of children and adolescents in Izmir/Turkey with the toxicological analysis results obtained from Addiction Toxicology Laboratory.Urine and blood samples of 4524 cases at and under the age of 18 years coming from various departments to the laboratory in 2015-2016 were analyzed by enzymatic immunoassay. Information and analysis results of the cases were obtained by retrospective analysis of the hospital system.83,3% of the cases were male and the mean age was 16,69 ± 1,63. Alcohol and/or substance use was determined in 13,2% of the cases. Among the cases with positive results of analysis, cannabis (33%) was mostly detected and was followed by amphetamine type stimulants (ATS, 15%), polysubstance use (15%) and alcohol (13%). While cannabis, polysubstance use and ATS were the most common in male, ethyl alcohol, ATS and benzodiazepine were mostly detected in female. There was a significant increase in the substance use rate in 2016 compared to the previous year.A substance use profile was obtained through drug testing in adolescents who are in the risk group for substance use. In this context, our data will be indicative for the development of new and more effective preventive strategies targeting children and adolescents.

Investigating the Effects of a Synthetic Cannabinoid on the Pathogenesis of Leukemia and Leukemic Stem Cells: A New Therapeutic Approach.

The popularity and usage of synthetic cannabinoids (SCs) are increasing due to their easy accessibility and psychoactive effects worldwide. Studies on cannabinoids on leukemic stem cells (LSC) and hematopoietic stem cells (HSCs), which are the precursors of leukemia cells, generally depend on the natural cannabinoid delta-9-THC. As there is only a limited number of studies focusing on the results of SC applications, the reflections upon LSCs have to be clarified. In this study, biological responses and antileukemic effects of JWH-018-one of the first produced and widely used SCs-were evaluated upon leukemia cells. Whether JWH-018 exhibited a preventive effect on both leukemic and HSCs was evaluated by presenting a therapeutic approach for the first time in the literature. Cells were analyzed in case of cell proliferation, apoptosis, and transcriptional expression profiling of some significant JAK/STAT and AKT/mTOR pathways, apoptotic, cell cycle regulation, and epigenetic chromatin remodeling-related genes following JWH-018 treatment. In conclusion, however, further studies are still needed upon both HSCs and LSCs to illuminate the effects of SCs on leukemogenesis on chronic myeloid leukemia (CML) more clearly; we consider that the JWH-018 can provide a therapeutic effect on the pathogenesis of leukemia and particularly upon LSCs and SCs might have therapeutic potential in addition to current therapy.

Antimicrobial synergism and antibiofilm activities of Pelargonium graveolens, Rosemary officinalis, and Mentha piperita essential oils against extreme drug-resistant Acinetobacter baumannii clinical isolates.

Rosemary officinalis L., Pelargonium graveolens L., and Mentha piperita L., essential oils are used by complementary medicine specialists simultaneously with traditional antibiotics for treatment purposes. The chemical composition of essential oils was analyzed by the gas chromatography-mass spectrometry method. In vitro antibacterial and antibiofilm activities of the essential oils were tested against extreme drug-resistant (XDR) colistin-resistant and colistin susceptible Acinetobacter baumannii clinical strains. The synergistic activities between essential oils and colistin antibiotics were investigated by the checkerboard method. The highest antibacterial effect was detected in mint essential oil (2.5-5 µl/ml), followed by pelargonium essential oil (5-20 µl/ml) and rosemary essential oil (5-20 µl/ml). The combination of rosemary essential oil or pelargonium essential oil with colistin showed strong synergistic activity in most of the bacterial strains tested (fractional inhibitory concentration index ≤ 0.5; synergy). As a result of the combination of mint essential oil and colistin, an indifferent effect was observed in only two bacterial strains, and other strains could not be evaluated. No antagonistic effects were observed in any of the tested essential oils. As a result of the effectiveness of the combination, the minimum inhibitory concentration (MIC) values of colistin in XDR-A. baumannii clinical isolates decreased 2-32 fold. Additionally, the sub-MIC concentration of essential oils exhibited an inhibitory effect (48-90%) against the biofilm layer of tested A. baumannii strains.

Determination of synthetic cannabinoids in randomly urine samples collected from probationers in Turkey.

Synthetic cannabinoids are a significant public health and safety problem that complicates drug tests with their ever-changing structures in our country and worldwide. The fact that most synthetic cannabinoids cannot be detected in biological samples by routine drug of abuse screening tests also causes an increase in the use of these substances in return. In this study, 500 urine samples of randomly selected probationers, analyzed with an enzymatic immunoassay test at Ege University Institute of Drug Addiction, Toxicology and Pharmaceutical Sciences (BATI) and tested negative, were then selected for retrospective analysis. Synthetic cannabinoids and their metabolites were quantitatively scanned in the collected urine samples via the liquid-liquid extraction method with the LC-MS/MS. Of the 500 studied urine samples, 108 (21.6%) were positive for 20 synthetic cannabinoids and their metabolites. The two most detected synthetic cannabinoids were 5F-NPB-22 (58%) and (S)-AB-FUBINACA (36%), and their mean concentrations were 72.94 ±â€¯47.51 ng/mL and 5.84 ±â€¯14.7 ng/mL, respectively. These results were also compared with national statistics from the general population. It resulted that immunoassay screening tests used in this study were insufficient, and urine samples should be studied in clinical and forensic cases with a validated chromatographic method.

New anti-viral drugs for the treatment of COVID-19 instead of favipiravir.

The SARS-CoV-2 virus is a major problem in the world right now. Currently, all the attention of research centers and governments globally are focused on the investigation of vaccination studies and the discovery of small molecules that inhibit the SARS-CoV-2 virus in the treatment of patients. The goal of this study was to locate small molecules to be used against COVID19 instead of favipiravir. Favipiravir analogues were selected as drug candidates from the PubChem web tool. The RNA dependent RNA polymerase (RdRp) protein was selected as the target protein as favipiravir inhibits this protein in the human body. Initially, the inhibition activity of the studied compounds against RdRp of different virus types was investigated. Then, the inhibition properties of selected drug candidates and favipiravir were examined in detail against SARS-CoV-2 RdRp proteins. It was found that 2-oxo-1H-pyrazine-3-carboxamide performed better than favipiravir in the results of molecular docking, molecular mechanics Poisson-Boltzmann surface area (MM-PSBA) calculations, and ADME analyses.Communicated by Ramaswamy H. Sarma.

Peripheral TREM2 mRNA levels in early and late-onset Alzheimer disease's patients.

'Triggering receptor expressed on myeloid cells 2' (TREM2) gene is involved in Alzheimer's disease (AD) and TREM2 mRNA expression is known to be increased in the peripheral blood cells of AD patients. In this study, we examined the expression levels of TREM2 mRNA in peripheral leukocytes of early and late-onset AD patients. We have also investigated the effect of the presence of APOE ε4 allele on TREM2 expression. TREM2 mRNA expression was analyzed in 30 early-onset AD (EOAD) patients, 38 late-onset AD (LOAD) patients, and in their age-matched controls by using quantitative real-time polymerase chain reaction. TREM2 levels in LOAD patients were higher than EOAD. Also, in elderly controls significantly higher TREM2 levels were found compared with young controls. Moreover, APOE ε4 carriers in LOAD patients exhibited significantly higher TREM2 expression levels than APOE ε4 non-carriers and elderly controls. Also, correlation analysis showed that TREM2 mRNA expression was increased by age. The differential expression of TREM2 mRNA levels between EOAD and LOAD patients might be independent of the AD disease status and results from an age-related increase in TREM2 expression. In LOAD patients, increased age and the presence of APOE ε4 allele further increase TREM2 expression. Taken together, we can suggest that age is a factor that increases TREM2 expression, and TREM2 and APOE ε4 may interact together in the pathogenesis of LOAD.