Biofilm Formation and Antibiotic Resistance Profiles in Carbapenemase-Producing Gram-Negative Rods-A Comparative Analysis between Screening and Pathological Isolates.

(1) Background: Carbapenem-resistant (CR) bacteria pose a significant global public health challenge due to their ability to evade treatment with beta-lactam antibiotics, including carbapenems. This study investigates the biofilm-forming capabilities of CR clinical bacterial isolates and examines the impact of serum on biofilm formation. Additionally, the study evaluates the resistance profiles and genetic markers for carbapenemase production. (2) Methods: Bacterial isolates were collected from the microbiology laboratory of Mures County Clinical Hospital between October 2022 and September 2023. Pharyngeal and rectal swabs were screened for carbapenem-resistant bacteria using selective media. Lower respiratory tract samples were also analyzed for CR Gram-negative bacteria. The isolates were tested for their ability to form biofilms in the presence and absence of fetal bovine serum at 24 and 48 h. Carbapenemase production was detected phenotypically and confirmed via PCR for relevant genes. (3) Results: Out of 846 screened samples, 4.25% from pharyngeal swabs and 6.38% from rectal swabs tested positive for CR bacteria. Acinetobacter baumannii and Klebsiella pneumoniae were the most common species isolated. Biofilm formation varied significantly between clinical isolates and standard strains, with clinical isolates generally showing higher biofilm production. The presence of serum had no significant effect on biofilm formation in Klebsiella spp., but stimulated biofilm formation for Acinetobacter spp. Carbapenemase genes blaKPC, blaOXA-48-like, and blaNDM were detected in various isolates, predominantly in Klebsiella spp., but were not the main determinants of carbapenem resistance, at least in screening isolates. (4) Conclusions: This study highlights the variability in biofilm formation among CR clinical isolates and underscores the differences between the bacteria found as carriage versus infection. Both bacterial species and environmental factors variably influence biofilm formation. These insights are crucial for the development of effective treatment and infection control strategies in clinical settings.

Correlation between Coronary Artery Disease and Non-Alcoholic Fatty Liver Disease Using Computed Tomography Coronary Calcium Scans.

INTRODUCTION: Concerns about how non-alcoholic fatty liver disease (NAFLD) might contribute to the development of cardiovascular disease (CVD) have grown as the importance of NAFLD and its relationship to the metabolic syndrome has grown. The purpose of this cross-sectional retrospective is to investigate potential correlations between hepatic steatosis in liver segments seen when measuring calcium score and the presence of atherosclerotic CAD (coronary artery disease). METHODS: Two hundred patients (mean age, 57 years±10) who underwent coronary cardiac computed tomography (CT) scans were included. CT scans were analysed to assess the attenuation of liver parenchyma and the coronary artery calcification (CAC). RESULTS: Age, gender, body mass index (BMI) and CAC score were significantly associated with hepatic steatosis. Among all patients, CAC score (r=-0.31, p<0.0001), and BMI (r=-0.40, p<0.0001) had a moderate negative correlation with the values of liver attenuation. BMI (OR: 1.109, p=0.001), CAC score (OR: 1.629, p<0.001), and age (OR: 1.050, p<0.001) were found to be independent predictors of hepatic steatosis through logistic regression. CONCLUSIONS: A statistically significant correlation between CAC score and the presence of NAFLD as evaluated by non-contrast-enhanced CT was demonstrated. BMI, CAC score, and age were identified as independent predictors of hepatic steatosis.

Uncovering the Resistance Mechanisms in Extended-Drug-Resistant Pseudomonas aeruginosa Clinical Isolates: Insights from Gene Expression and Phenotypic Tests.

(1) Background: The purpose of the study was to describe the activity of mex efflux pumps in Multidrug-Resistant (MDR) clinical isolates of Pseudomonas aeruginosa and to compare the carbapenem-resistance identification tests with PCR; (2) Methods: Sixty MDR P. aeruginosa were analyzed for detection of carbapenemase by disk diffusion inhibitory method, carbapenem inactivation method and Modified Hodge Test. Endpoint PCR was used to detect 7 carbapenemase genes (blaKPC, blaOXA48-like, blaNDM, blaGES-2, blaSPM, blaIMP, blaVIM) and mcr-1 for colistin resistance. The expression of mexA, mexB, mexC, mexE and mexX genes corresponding to the four main efflux pumps was also evaluated; (3) Results: From the tested strains, 71.66% presented at least one carbapenemase gene, with blaGES-2 as the most occurring gene (63.3%). Compared with the PCR, the accuracy of phenotypic tests did not exceed 25% for P. aeruginosa. The efflux pump genes were present in all strains except one. In 85% of the isolates, an overactivity of mexA, mexB and mostly mexC was detected. Previous treatment with ceftriaxone increased the activity of mexC by more than 160 times; (4) Conclusions: In our MDR P. aeruginosa clinical isolates, the carbapenem resistance is not accurately detected by phenotypic tests, due to the overexpression of mex efflux pumps and in a lesser amount, due to carbapenemase production.

The Association of Nonalcoholic Fatty Liver Disease With Neutrophil-to-Lymphocyte Ratio and Neutrophil-Percentage-to-Albumin Ratio.

Background and objective Nonalcoholic fatty liver disease (NAFLD) is closely linked to metabolic syndrome, leading to consequences related to dyslipidemia, endothelial dysfunction, type 2 diabetes, and obesity. Due to a limited understanding of the factors contributing to the progression of NAFLD, predicting clinical outcomes in patients remains challenging. In light of this, this study aimed to evaluate the association between the occurrence of NAFLD and the neutrophil-percentage-to-albumin ratio (NPAR) as well as the neutrophil-to-lymphocyte ratio (NLR). Methods A total of 115 adult patients (mean age: 58 ± 12.5 years; 55.65% male) who underwent abdominal non-contrast-enhanced CT scans were included in the study. The analysis of CT scans was conducted to assess the attenuation values of liver parenchyma. Results  There was a statistically significant difference in terms of gamma-glutamyl transpeptidase (GGT), triglyceride (TG), albumin, and NPAR between individuals with and without hepatic steatosis (GGT p<0.0001, TG p=0.0006, albumin p<0.0001, NPAR p=0.001). However, NLR values between the two groups did not show any statistical differences. NPAR (r=-0.27, p=0.0029) had a weak inverse correlation with liver attenuation value, which is expressed in Hounsfield units (HU). Conclusions Significant differences were observed in GGT, TG, albumin, and NPAR levels between individuals with and without hepatic steatosis. An inverse correlation between NPAR and liver attenuation values was also observed.

Correlation between CT Abdominal Anthropometric Measurements and Liver Density in Individuals with Non-Alcoholic Fatty Liver Disease.

Background: With a growing frequency, nonalcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease worldwide. NAFLD has a strong correlation with other metabolic disorders, such as obesity, particularly abdominal obesity, even though the underlying causes or risk factors are not entirely understood. This study aims to investigate correlations between abdominal anthropometric measurements and the presence and intensity of liver steatosis as assessed by unenhanced computed tomography (CT). Methods: One hundred and nineteen patients (male/female, 66/53; mean age 54.54 +/- 12.90 years) underwent abdominal non-contrast-enhanced CT. CT images were examined to determine the attenuation of liver parenchyma, subcutaneous fat depth, and waist circumference (WC). Results: Among all patients, WC (r = -0.78, p < 0.0001), infraumbilical subcutaneous fat thicknesses (r = -0.51, p < 0.0001), right paraumbilical subcutaneous fat thicknesses (r = -0.62, p < 0.0001), and left paraumbilical subcutaneous fat thicknesses (r = -0.53, p < 0.0001) had a high inverse correlation with the liver attenuation values. The presence of T2D (OR: 2.40, p = 0.04), WC (OR: 11.45, p < 0.001), right paraumbilical (OR: 10.09, p < 0.001), left paraumbilical (OR: 2.81, p = 0.01), and infraumbilical (OR: 3.06, p = 0.007) were strongly independent predictors of NAFLD risk. Moreover, regarding the laboratory parameters, only the higher value of GGT (OR: 2.84, p = 0.009) is a predictor of NAFLD risk. Conclusions: Our data show that higher baseline values of all abdominal anthropometric measurements are correlated with liver attenuation and act as predictors of NAFLD risk.

Antibacterial Effect of 16 Essential Oils and Modulation of mex Efflux Pumps Gene Expression on Multidrug-Resistant Pseudomonas aeruginosa Clinical Isolates: Is Cinnamon a Good Fighter?

The purpose of the study was to describe the antimicrobial activity of 16 common essential oils (EOs) on multidrug-resistant (MDR) Pseudomonas aeruginosa clinical isolates, including the determination of the effects on mex efflux pumps gene expression. Seventy-two clinical isolates of P. aeruginosa collected between 2020-2022 were screened for susceptibility to EOs using Kirby-Bauer disk diffusion to identify potential candidates for future alternative therapies. The minimal inhibitory concentration (MIC) was further determined for the EO that proved antibacterial activity following the disk diffusion screening. Positive and negative controls were also used for method validation. Since cinnamon EO exhibited the best antimicrobial activity, it was further used to evaluate its influence on mex A, B, C, E, and X efflux pumps gene expression using real-time RT-PCR. Cinnamon EO inhibited all P. aeruginosa strains, followed by thyme EO (37.5%, n = 27) and lavender EO (12.5%, n = 9). The other EOs were less efficient. The MIC detection showed that cinnamon at a concentration of 0.05% v/v inhibited all MDR P. aeruginosa isolates. Thyme, turmeric, peppermint, basil, clove, and lavender EOs presented various results, most of them having activity at concentrations higher than 12.5% v/v. By studying the activity of cinnamon EO on mex efflux pumps, it was found that mexA and mexB (66.5%) were generally under-expressed. The remarkable results produced using the very low concentrations of cinnamon EO, with 100% antimicrobial activity against multi-, extended-, and pan- drug-resistant (MDR, XDR, PDR) P. aeruginosa clinical isolates, completed with the severe alteration of the RNA messaging system, supports its potential to be used as adjuvant treatment, with impact on therapeutic results.

Epidemiology, Evolution of Antimicrobial Profile and Genomic Fingerprints of Pseudomonas aeruginosa before and during COVID-19: Transition from Resistance to Susceptibility.

BACKGROUND: The purpose of the study was to describe the epidemiological implication of Pseudomonas aeruginosa between 2017-2022 in a tertiary hospital from Romania, including the molecular fingerprinting of similar phenotypic strains (multidrug-resistant isolates), which would have an important health impact. The study also describes the resistance profile of P. aeruginosa before and during COVID-19, which might bring new information regarding the management of antibiotic treatments. MATERIALS AND METHODS: Information regarding wards, specimen types, species, and antibiotic resistance profile of 1994 strains of Pseudomonas spp. Isolated over a period of 6 years in Mures Clinical County Hospital, Romania, was collected from the WHONET database. From 50 multidrug-resistant isolates, molecular fingerprinting was performed by Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) to prove the potential clonal distribution. RESULTS: A number of 1994 Pseudomonas spp. were isolated between 2017-2022, from which P. aeruginosa was the most frequent species, 97.39% (n = 1942). P. aeruginosa was most frequently isolated in 2017 (n = 538), with the dermatology department as the main source, mainly from pus secretion. A drop in the harvesting rate was noted in 2020 due to COVID-19 restrictions. Regarding the resistance profile, there are a few modifications. The susceptibility of P. aeruginosa to carbapenems, piperacillin-tazobactam, and amikacin suffered alterations before and during COVID-19. The molecular fingerprinting showed three P. aeruginosa clusters, including strains with 80-99% similarity.