Exploring the Link between Inflammatory Biomarkers and Adipometrics in Healthy Young Adults Aged 20-35 Years.

Obesity and aging are associated with an inflammatory state, which represents the common background for a wide range of diseases. This study aims to explore the correlation between hsCRP, IL-1ß, IL-6, TNF-α, IFN-γ, and white blood cell count (WBC) and adipometrics (arm, waist, and hip circumferences: AC, WC, HC; total body fat mass: TBFM, visceral fat level: VFL, body mass index: BMI; waist/hip ratio: WHR; waist/height ratio: WHtR) in young and healthy adults aged 20-35 years old. The subjects were divided by BMI into the overweight/obesity (OW/OB) group and normal weight (NW) group, and by hsCRP level into Group 1 (<1 mg/L), Group 2 (≥1-2.99 mg/L), and Group 3 (≥3 mg/L). The concentration of all inflammatory biomarkers was significantly higher in the OW/OB group compared to the NW group, with the exception of IL-1ß. Significant positive correlations were found between hsCRP, TNF-α, WBC, and all adipometrics; between IL-6 and WHR, WHtR, BMI, TBFM, and VFL; and between IFN-γ and HC, BMI, and TBFM. IL-1ß correlates positively with WHR and VFL. In Groups 1-3, all the differences between the adipometrics showed significant differences. Subclinical inflammation persists in association with being overweight and obese in healthy young adults aged 20-35 years old.

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.

Candida spp. DNA Extraction in the Age of Molecular Diagnosis.

The standard procedure for the detection of candidemia is blood culture, a method that might require 3-5 days for a positive result. Compared with culturing, molecular diagnosis techniques can provide faster diagnosis. The current paper aimed to present the main strengths and constraints of current molecular techniques for Candida spp. DNA extraction, analyzing their efficiency from a time, price, and ease of usage point of view. A comprehensive search was conducted using the PubMed NIH database for peer-reviewed full-text articles published before October 2022. The studies provided adequate data on the diagnosis of the infection with the Candida spp. DNA extraction is a relevant step in yielding pure qualitative DNA to be amplified in molecular diagnostic techniques. The most used fungal DNA extraction strategies are: mechanical (bead beating, ultrasonication, steel-bullet beating), enzymatic (proteinase K, lysozyme, lyticase), and chemical extraction (formic acid, liquid nitrogen, ammonium chloride). More clinical studies are needed to formulate adequate guidelines for fungal DNA extraction as the current paper highlighted discrepancies in the reported outcome.

Potential Use of Quercus dalechampii Ten. and Q. frainetto Ten. Barks Extracts as Antimicrobial, Enzyme Inhibitory, Antioxidant and Cytotoxic Agents.

The bark resulted as a by-product after different Quercus sp. processing is a valuable alternative source of phenolic compounds (mainly tannins). Hence, the focus of the present work was to obtain eight extracts from the rhytidome of the less-studied Q. dalechampii and Q. frainetto and characterize them in terms of chemical and bioactive profiles. Ultrasound and microwave-assisted extraction methods were used for the preparation of the extracts. Total phenolic and tannin contents were assessed through classic spectrophotometric methods, while several individual phenolic compounds were identified and quantified using UPLC-PDA. Antioxidant, enzyme-inhibitory, antibacterial, and cytotoxic activities were tested using in vitro assays; additionally being evaluated was the ability of the extracts to inhibit the adherence of MRSA to suture wires. The UPLC analysis confirmed the presence of gallic acid, catechin, taxifolin, vanillic acid, epicatechin, and caffeic acid. The results showed that tested extracts were able to exert cytotoxic effects, at 6% and 3% concentrations, on confluent cells. The tested solutions inhibit α-glucosidase activity and the antibacterial potential suggested a mild to moderate effect against the Gram-positive strains. Overall, the obtained results revealed rich phenolic and tannin contents for the extracts obtained from both species through microwave-assisted extraction, probably responsible for their mild antibacterial and cytotoxic effects.

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.

Biological Activity of Bark Extracts from Northern Red Oak (Quercus rubra L.): An Antioxidant, Antimicrobial and Enzymatic Inhibitory Evaluation.

The northern red oak (Quercus rubra L.) is an ornamental oak species native to eastern America, being an invasive species in Europe, with increasing coverage. The aim of this work was to evaluate the biological potential of red oak bark extracts. Aqueous and ethanolic preparations were obtained by two extraction methods: ultrasonic-assisted extraction (UAE) and microwave assisted extraction (MAE). The total phenolic and tannin contents were measured using spectrophotometric methods. The antioxidant activity was evaluated by two complementary methods (DPPH and ABTS). Antimicrobial potential was tested against five bacteria and three Candida species, and the effect on biofilm formation and synergism with gentamicin was also evaluated. Finally, enzyme inhibitory properties were assessed for α-glucosidase, tyrosinase, and acetylcholinesterase. The results indicated a higher phenolic content for the extracts obtained through MAE, while UAE bark extracts were rich in tannins. All the extracts exhibited antioxidant, anti-glucosidase, and anti-tyrosinase activity, while the antibacterial potential was mostly observed for the MAE extracts, especially against S. aureus, C. parapsilopsis, and C. krusei; inhibition of biofilm formation was observed only for MRSA. These findings show that the red oak bark might be an important source of bioactive compounds with antioxidant and antimicrobial properties.

Serogroups and genetic diversity of diarrheagenic strains of Escherichia coli: a retrospective study.

INTRODUCTION: Diverse serogroups of Escherichia coli cause sporadic cases and outbreaks of diarrhea among children. Our study aimed to evaluate the serogroups of diarrheagenic strains of E. coli that cause diarrheal disease in children under two years old, and clarify if the cases were sporadic or outbreaks. METHODOLOGY: The retrospective study included 130 strains of pathogenic E. coli, isolated from children who were less than two years of age, and had diarrheal disease, between May 2016 and July 2019. The study was conducted in the Bacteriology Laboratory (County Clinical Hospital, Mureș, Romania). The 130 strains were sero-grouped using polyvalent and monovalent O antisera. Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) was performed to evaluate the similarity between different E. coli strains, and a simplex polymerase chain reaction (PCR) was performed to detect the presence of the hlyA gene that is specific to the enterohemorrhagic strains. RESULTS: After agglutination with polyvalent O antisera, slightly more than half of the strains (50.77%) were sero-grouped as Shiga toxin-producing E. coli (STEC), and the rest of the strains belonged to the Enteropathogenic Escherichia coli (EPEC) serogroups. Serogroup O157 was the most frequently identified (16.51% of the total number of typeable strains), and one strain was positive for hlyA. ERIC-PCR revealed a high diversity of strains, with an overall 50% similarity. CONCLUSIONS: STEC serogroups were the most common strains causing diarrheal disease, and O-157 was the dominant serogroup identified. The strains included in our study presented high genetic diversity, suggesting that most of the cases were sporadic.

Antifungal and anti-virulence activity of six essential oils against important Candida species - a preliminary study.

Opportunistic infections with Candida species are becoming more problematic, considering their increasing virulence and resistance to antifungal drugs. Aim: To assess the antifungal and anti-virulence activity of basil, cinnamon, clove, melaleuca, oregano and thyme essential oils (EOs) on five Candida species (C. albicans, C. auris, C. krusei, C. parapsilosis and C. guillermondii). Methods: The MIC, growth rate, antibiofilm activity, regulation of gene expression (ALS3, SAP2, HSP70) and germ-tube formation were evaluated by specific methods. Results: Most EOs inhibited Candida species growth and reduced the expression of some virulence factors. Cinnamon and clove EO showed the most significant inhibitory effects. Conclusions: The tested EOs are promising agents for facilitating the management of some Candida infections.

Optimization of Microwave Assisted Extraction Conditions to Improve Phenolic Content and In Vitro Antioxidant and Anti-Microbial Activity in Quercus cerris Bark Extracts.

The species of the Quercus genus, including Quercus cerris L., are widely used and their wood represents a valuable material utilized for various purposes. The intense industrial processing of wood results in a considerable amount of poorly-used natural by-products, such as bark, and a loss of potentially useful raw materials. Thus, the aim of our study was to evaluate the phytochemical characteristics and potential biological activities of Quercus cerris bark extracts obtained by optimizing the parameters of microwave assisted extraction (MAE). The optimum conditions for MAE were determined using a design of experiments (DoE) model, which set the experimental variables (irradiation time and microwave power) and their values. Aqueous and hydroalcoholic extractions were performed and the optimum parameters of extraction were determined for both solvents. The total polyphenolic and tannin contents were determined. The biological activities representative of antioxidant capacity were determined using two free radical scavenging methods, the DPPH and ABTS methods, and the antibacterial activity was assessed with the microdilution method. The results showed different optimal extraction conditions for aqueous (30 min at 850 W) and hydroalcoholic (18 min at 650 W) extracts. A higher yield of total polyphenols was observed in the hydroalcoholic bark extract (403.73 ± 7.35 mg gallic acid equivalents/g dried weight); however a lower level of tannins was registered in comparison to the aqueous extract. In addition, both extracts exerted high antioxidant activities, with the aqueous extracts having a stronger inhibitory effect against the DPPH radical. Moreover, the extracts exhibited antibacterial activity against the tested bacterial strains, especially against the Gram-positive strains and Klebsiella pneumoniae, with the hydroalcoholic extracts being more efficient overall. To conclude, the optimized MAE was an efficient method to extract phytochemical compounds with potential biological effects from Quercus cerris bark.

Candida auris and other phylogenetically related species - a mini-review of the literature.

The lesser-known non-albicans Candida species are often overlooked and difficult to diagnose in most microbiology laboratories. Candida auris, a relatively newly discovered species, is responsible for outbreaks in various geographical regions. Because of its increased resistance to antifungal drugs, C. auris is responsible for hard-to-treat infections and its pathogenicity is still incompletely elucidated. Non-albicans species phylogenetically related to C. auris, like the C. haemulonii complex might also play a role in human pathology. The current summary of the literature presents the emergence, virulence, laboratory identification, and molecular mechanisms responsible for antifungal resistance of emergent rare non-albicans Candida species.

Candida Spp. in Lower Respiratory Tract Secretions - A Ten Years Retrospective Study.

INTRODUCTION: Lower respiratory tract secretions (LRTS) like sputum and tracheal aspirates are frequently sent to the microbiology laboratory from patients with various respiratory pathologies. Improper collection techniques can lead to false-positive results, resulting in improper therapy. AIM OF THE STUDY: To determine the percentage of contaminated samples sent to the microbiology laboratory, to establish the prevalence of Candida spp. in non-contaminated samples and therefore, the presence of Candida spp. originating in lower respiratory tract infections. Material and. METHODS: A 10-year data survey was conducted to assess the differences in Candida prevalence from contaminated versus non-contaminated samples, assessed and categorised by Bartlett grading system, and to emphasise the importance of quality control for potentially contaminated samples. The data were analysed according to gender, age, referring departments, and the species of Candida. For the statistical analysis, Kruskal-Wallis and Fisher tests were used, and the alpha value was set for 0.5. RESULTS: The prevalence of Candida spp. in all analysed samples was 31.60%. After excluding the contaminated samples, the actual prevalence was 27.66%. Of all sputum samples, 31.6% were contaminated. Patients aged more than 40 years old were more prone to provide contaminated sputum samples. C. albicans is more prevalent in non-contaminated sputum samples. In both sputum and tracheal aspirates, the chances of identifying a single species are higher than the chances of identifying multiple species. CONCLUSIONS: The study emphasises the importance of assessing the quality of sputum samples because of the high number of improperly collected samples sent to the microbiology laboratory.

Silver Nanoparticles Biosynthesized with Spruce Bark Extract-A Molecular Aggregate with Antifungal Activity against Candida Species.

Due to their high content of biomolecules, combined with silver's well known antimicrobial potential, silver nanoparticles biosynthesized using spruce bark (AgNP SBEs) demonstrate antibacterial and antioxidant activity, making them a versatile option for developing new antimicrobial agents that might be used for medical treatment or as adjuvants for the classical agents. This study aims to analyze if silver nanoparticles (AgNPs) mediated by spruce bark extract (SBE) and silver salts (AgNP SBE Acetate, AgNP SBE Nitrate) presents antifungal activity against five different Candida spp., synergistic activity with fluconazole, and if they interact with some virulence factors of C. albicans. AgNP SBEs presented MICs (minimum inhibitory concentrations) for all the five tested Candida spp. AgNP SBEs inhibited the growth of C. parapsilosis, C. krusei, and C. guilliermondii, exerted synergistic activity with fluconazole for C. parapsilosis and C. guilliermondii, and inhibited biofilm production for C. albicans, C. auris, and C. guilliermondii. MICs of AgNP SBE Acetate significantly inhibited the production of germ tubes of C. albicans. The expression of C. albicans SAP2 gene was down-regulated by the short-time treatment with MICs of AgNP SBE Acetate, while ALS3 and HSP70 genes were up-regulated by the AgNPs MICs. These results emphasize the potential of using the AgNP SBEs as treatments/adjuvants options, not only against the redundant C. albicans but also for the non-albicans Candida species (which are not as frequently involved in human pathologies, but, sometimes, can be more aggressive).

In Vitro Antifungal Activity of Silver Nanoparticles Biosynthesized with Beech Bark Extract.

Biosynthesis is a green method for the synthesis of silver nanoparticles (AgNPs). This study aimed to assess the antifungal activity of two silver nanoparticle solutions, synthesized using beech bark extract (BBE) and acetate and nitrate silver salts (AgNP Acetate BBE and AgNP Nitrate BBE), their influence on biofilm production, their potential synergistic effects with fluconazole, on different Candida spp., and their influence on virulence factors of C. albicans (germ tube production, gene expression for ALS3, SAP2, HSP70). Both the AgNP BBEs presented different minimum inhibitory concentrations for all the studied Candida spp., but biofilm production was inhibited only for C. albicans and C. guilliermondii. The growth rates of all the studied Candida spp. were inhibited in the presence of both AgNP BBEs, except for C. auris. Synergistic activity was observed for C. parapsilosis and C. guilliermondii, for different combinations of fluconazole with both the AgNP BBEs. The germ tube production of C. albicans was slightly inhibited by the AgNP BBEs. Only AgNP Acetate BBE was able to down-regulate the expression of SAP2. Overall, we can conclude that, even if more studies are necessary, AgNPs synthesized with beech bark extract might be an interesting alternative to classic antifungal treatments.

Hemolysin-Producing Strains among Diarrheagenic Escherichia coli Isolated from Children under 2 Years Old with Diarrheal Disease.

Even if serotyping based on O antigens is still routinely used by most laboratories for the detection of diarrheagenic Escherichia coli, this method can provide false-positive reactions, due to the high diversity of O antigens. Molecular methods represent a valuable tool that clarifies these situations. In the Bacteriology Laboratory of Mureș County Hospital, between May 2016 and July 2019, 160 diarrheagenic E. coli strains were isolated from children under 2 years old with diarrheic disease. The strains were identified as Shiga toxin-producing E. coli (STEC)/enteropathogenic Escherichia coli (EPEC) via agglutination with polyvalent sera. STEC strains were serotyped using monovalent sera for serogroup O157. Simplex PCR was performed on the strains to determine the presence of the hlyA gene, and, for the positive ones, the hemolytic activity was tested. Antibiotic susceptibility of the identified diarrheagenic E. coli strains was also investigated. STEC strains were the most frequently identified (49.1%), followed by EPEC (40.2%). The hlyA gene was identified in 12 cases, representing 18.2% of the STEC strains. Even if the extended-spectrum ß-lactamase (ESBL)-producing strains represented only 10%, a relevant percentage of multidrug-resistant (MDR) strains (24%) was identified.

Candida and Candidiasis-Opportunism Versus Pathogenicity: A Review of the Virulence Traits.

One of the most important questions in microbiology nowadays, is how apparently harmless, commensal yeasts like Candida spp. can cause a rising number of infections. The occurrence of the disease requires firstly the attachment to the host cells, followed by the invasion of the tissue. The adaptability translates into a rapid ability to respond to stress factors, to take up nutrients or to multiply under different conditions. By forming complex intracellular networks such as biofilms, Candida spp. become not only more refractive to antifungal therapies but also more prone to cause disease. The inter-microbial interactions can enhance the virulence of a strain. In vivo, the fungal cells face a multitude of challenges and, as a result, they develop complex strategies serving one ultimate goal: survival. This review presents the virulence factors of the most important Candida spp., contributing to a better understanding of the onset of candidiasis and raising awareness of the highly complex interspecies interactions that can change the outcome of the disease.

New perspectives on the nutritional factors influencing growth rate of Candida albicans in diabetics. An in vitro study.

BACKGROUND: The link between Candida albicans and diabetes mellitus is well-acknowledged, but incompletely elucidated. OBJECTIVES: The purpose of this study is to assess the growth rate of C. albicans (CA) in the presence of different concentrations of glucose and fructose, two of the main pathophysiologic and nutritionally relevant sugars in diabetic patients, in order to obtain a better understanding of the nutrient acquisition strategy and its possible relation to the hyperglycemic status of diabetic patients. METHODS: The effects of different concentrations of glucose and fructose (1000 mg%, 500 mg%, 250 mg% and 100 mg% w/v) on the growth rate of CA have been studied by flow-cytometry. FINDINGS: We found that glucose concentration is directly related to CA growth, which may be linked to the frequent yeast infections that occur in non-controlled diabetic patients; we also show that fructose inhibits CA growth rate. MAIN CONCLUSIONS: As a consequence of our hypothesis, the study demonstrates that fructose-containing food may prevent the development of candidiasis, at least in oral sites.

New perspectives on the nutritional factors influencing growth rate of Candida albicans in diabetics. An in vitro study

BACKGROUND The link between Candida albicans and diabetes mellitus is well-acknowledged, but incompletely elucidated. OBJECTIVES The purpose of this study is to assess the growth rate of C. albicans (CA) in the presence of different concentrations of glucose and fructose, two of the main pathophysiologic and nutritionally relevant sugars in diabetic patients, in order to obtain a better understanding of the nutrient acquisition strategy and its possible relation to the hyperglycemic status of diabetic patients. METHODS The effects of different concentrations of glucose and fructose (1000 mg%, 500 mg%, 250 mg% and 100 mg% w/v) on the growth rate of CA have been studied by flow-cytometry. FINDINGS We found that glucose concentration is directly related to CA growth, which may be linked to the frequent yeast infections that occur in non-controlled diabetic patients; we also show that fructose inhibits CA growth rate. MAIN CONCLUSIONS As a consequence of our hypothesis, the study demonstrates that fructose-containing food may prevent the development of candidiasis, at least in oral sites.