Biochemical Tests Research Articles

EFFECT OF PENICILLIUM SPECIES ON THE ANTIBIOTIC RESISTANCE PROFILE OF ALCALIGENES FAECALIS.

Infectious diseases due to antibiotic resistant pathogens are a global public health problem. This study aimed at determining the potential effect of bacterial-fungal interaction on the antibiotic susceptibility profile of Alcaligenes faecalis. Alcaligenes faecalis was isolated from water samples. The isolate was identified using the conventional biochemical tests and the 16S rRNA molecular sequencing technique. Additionally, Penicillium species was isolated and identified based on colony morphological characteristics and microscopic features. Standardized isolates were co-cultured in broth medium. Antibiotic susceptibility evaluation of the Alcaligenes faecalis from the co-culture and the original Alcaligenes faecalis was carried out using the Kirby bauer disk diffusion method. The antibiotic susceptibility profile of Alcaligenes faecalis before and after co-culture remained largely unchanged except in the case of chloramphenicol, where the isolate showed reduced susceptibility. Molecular analysis of resistance gene revealed the absence of tested gene encoding antibiotic resistance, including the streptomycin resistance (str) genes (stra and strb) and the erythromycin resistance methylase (erm) gene. The result of this study showed that there is a minimal influence of Penicillium cultures on the susceptibility of A. faecalis. Further research involving a wide spectrum of microorganisms and their interactions should be conducted to acquire a thorough understanding of the influence of microbial interactions on antibiotic susceptibility profiles in order to pave way for novel strategies to combat antimicrobial resistance.

Prevalence and antimicrobial-resistant patterns of Pseudomonas aeruginosa among burn patients attending Yekatit 12 Hospital Medical College in Addis Ababa, Ethiopia.

Burns are one of the most common forms of trauma globally. P. aeruginosa plays a prominent role as an etiological agent among burn patients. There is a paucity of information about the prevalence and antimicrobial resistance patterns of P. aeruginosa among burn patients in Ethiopia. Hence, this study was designed to assess the prevalence and antimicrobial-resistant patterns of P. aeruginosa among burn patients attending Yekatit 12 Hospital Medical College in Addis Ababa, Ethiopia. Hospital-based cross-sectional study was conducted at Yekatit 12 Hospital Medical College among burn patients from November 2020 to April 2021. Identification of P. aeruginosa was performed using Culture, Biochemical tests, and, Gram staining. Antimicrobial resistance testing was done using the Kirby-Bauer disc diffusion method. Logistic regression was computed to determine associated factors. From 210 burn wound cultures, 27 (12.86%) were found positive for P. aeruginosa. All the isolates showed greater than 70% susceptibility to the tested antibiotics except Gentamycin, Ceftazidime, and, Ciprofloxacin. In addition, 33.33% of P. aeruginosa isolates were multidrug-resistant. Admission type, Hospital stay time and Total body surface area (TBSA) had a statistically significant association (all with P-value <0.05) with the acquisition of P. aeruginosa infection. Overall, the prevalence of P. aeruginosa isolates among burn patients is almost 13%. Most P. aeruginosa isolates were sensitive to Imipenem, while they were most resistant to Gentamycin. One-third of P. aeruginosa were multidrug-resistant. This suggests the need to monitor the treatment of infection with the pathogen to limit the possibility of the emergence of multidrug-resistant isolates in burn centers.

Condition of the cardiovascular system in Wistar rats with experimental renal dysfunction

The aim: to evaluate functional and structural changes in blood vessels and myocardium in Wistar rats at different stages of the formation of experimental kidney dysfunction.Materials and Methods. Four groups of animals were studied. The first two groups consisted of rats subjected to 5/6 nephrectomy (NE). The period after NE is 2 or 4 months. The third and fourth groups included sham-operated (SO) animals of a similar observation period. After the end of the experiment, blood pressure was measured in rats, the left ventricular mass index (LVMI) was calculated, a biochemical blood test, histological, immunohistochemical and electron microscopic examination of the myocardium were performed, and the contractile activity of the portal vein was recorded for a period of 2 months (in vitro).Results. 2 months after NE, an increase in blood pressure, LVMI, and a decrease in the autorhythmic contractile activity of the portal vein were detected in rats. At the histological level, thickening of cardiomyocytes and arterial vessel walls and the presence of perivascular fibrosis were noted. After 4 months, in rats with NE, the increase in blood pressure, cardiomyocyte hypertrophy and perivascular fibrosis progressed. The thickness of cardiomyocytes was 14.1±3.11 μm, which was significantly greater than in the group with NE for 2 months (12.5±1.5 μm; p=0.008). At this period of observation, electron microscopic examination revealed deintegration of myofibrils, disruption of myofibril stacking and z-disk structure, and changes in the shape of mitochondria. 2 months after NE, an increase in the area of open capillaries was detected in rats (1902.8±202.9 μm²) compared to the corresponding LO animals (730.4±58.2 μm²; p=0.000). As renal dysfunction develops, the capillary area tends to increase (2139.1±396.5 μm²; p =0.120).Conclusion. In Wistar rats with renal dysfunction, changes were detected not only in the level of blood pressure, but also in the functional activity of the intravenous fluid, as well as in the structural components of the myocardium – cardiomyocytes, stroma and blood vessels. The increase in myocardial mass at the histological level was manifested by a significant increase in the thickness of cardiomyocytes, the volume of connective tissue, and the thickness of the wall of arterial vessels, reaching a maximum value at a longer period after NE.

Unraveling the functional characteristics of endophytic bacterial diversity for plant growth promotion and enhanced secondary metabolite production in Pelargonium graveolens

The rich diversity of microbial endophytic communities associated with plants, often referred to as the second genome, serves as a compelling illustration of efficient co-evolution. This noteworthy partnership plays a pivotal role in sustaining plant well-being and enhancing plant adaptability across diverse habitats. Therefore, examining the diversity of endophytic microbes associated with their particular host plant is valuable for gaining insights into the vast spectrum of plant-microbe interactions. The present experiments aimed at investigating the bacterial endophytic diversity in both root and shoot tissues of Pelargonium graveolens, employing culture dependent and culture independent high-throughput metagenomics approach. A total of 614 and 620 operational taxonomic units (OTUs), encompassing 291 and 229 genera, were identified in the shoot and root tissues of P. graveolens, respectively. Furthermore, the subsequent classification of OTUs revealed 15 highly abundant phyla, with Proteobacteria dominating both root and shoot tissues. Notably, an exceptionally high abundance of Firmicutes phyla was observed in the shoot compared to the root. Additionally, 30 bacterial endophytes from the root, stem, petiole, and leaves were isolated and molecularly characterized, unveiling a consistent pattern of diversity distribution between the root and shoot of P. graveolens. Upon screening all isolates for plant growth promoting traits, Pseudomonas oryzihabitans was found to be positive for major biochemical test like nitrogen fixation, phosphate solubilization etc. and on inoculation resulted in about two-fold increase in content of essential oil accompanied by a significant rise in the geraniol and citronellol content. Diving deep into the genetic constitution of P. oryzihabitans unveiled a substantial number of genes directly and indirectly contributing to the endophyte’s capability in colonizing host plants effectively. In summary, data obtained from metagenomics and culture dependent approaches including glass house trials suggest potential bacterial endophytes suitable for field applications for yield enhancement and in planta secondary metabolite enhancement investigations.

Assessment of the role of albuminuria and uromodulin in the early diagnosis of renal function disorders in some types of obesity phenotypes

The aim: to study of the relationship of biological markers reflecting damage of the kidneys with its functional state, as well as with clinical, metabolic, hormonal parameters and immunoinflammatory processes in obesity phenotypes.Patients and Methods: The study involved 224 obese Uzbek women who were divided into 2 groups according to the recommended criteria for metabolic syndrome: group 1 consisted of 133 women diagnosed with metabolic complicated obesity, average age 42.0±0.5; Group 2 consisted of 91 women diagnosed with metabolic healthy obesity, average age 41.7±0.7. The control group consisted of 45 healthy volunteers (women, mean age 43.2±0.8, body mass index ˂ 30 kg/m2, waist circumference less than 80 cm). In the observation groups, anthropometric indicators, blood pressure, biochemical tests and blood lipid spectrum, levels of leptin, insulin, cystatin C and uromodulin in the blood serum, gradations of microalbuminuria in urine were determined, the glomerular filtration rate was calculated for cystatin C and creatinine and the obtained indicators were compared.Results. In both obesity phenotypes, the amount of microalbuminuria and cystatin C significantly increased in group 1 compared to these indicators in group 2 and the control group, and the amount of uromodulin in the blood serum, on the contrary, decreased compared to the indicators in group 2 and control group (r ˂ 0.001) . An increase in the gradation of microalbuminuria and cystatin C and a decrease in the amount of uromodulin in the blood serum in both groups was expressed by a decrease in the estimated glomerular filtration rate, which was clearly expressed in group 1 (χ2 = 4.5, r = 0.034). Also, an increase in body mass index was characterized by an increase in cardiometabolic disorders, an increase in the gradation of microalbuminuria, a decrease in the level of uromodulin in the blood serum and cytokinemia (p ˂ 0.001).Conclusion. In both obesity phenotypes, an increase in the degree of obesity was manifested by an increase in the level of cardiometabolic risk, as well as an increase in subclinical kidney damage. It is considered appropriate to determine the amount of uromodulin and cystatin C in blood serum and assess the gradation of microalbuminuria in the early diagnosis of renal dysfunction in obesity.

Harnessing Next-Generation Sequencing as a Timely and Accurate Second-Tier Screening Test for Newborn Screening of Inborn Errors of Metabolism.

In this study, we evaluated the implementation of a second-tier genetic screening test using an amplicon-based next-generation sequencing (NGS) panel in our laboratory during the period of 1 September 2021 to 31 August 2022 for the newborn screening (NBS) of six conditions for inborn errors of metabolism: citrullinemia type II (MIM #605814), systemic primary carnitine deficiency (MIM #212140), glutaric acidemia type I (MIM #231670), beta-ketothiolase deficiency (#203750), holocarboxylase synthetase deficiency (MIM #253270) and 3-hydroxy-3-methylglutaryl-CoA lyase deficiency (MIM # 246450). The custom-designed NGS panel can detect sequence variants in the relevant genes and also specifically screen for the presence of the hotspot variant IVS16ins3kb of SLC25A13 by the copy number variant calling algorithm. Genetic second-tier tests were performed for 1.8% of a total of 22,883 NBS samples. The false positive rate for these six conditions after the NGS second-tier test was only 0.017%, and two cases of citrullinemia type II would have been missed as false negatives if only biochemical first-tier testing was performed. The confirmed true positive cases were citrullinemia type II (n = 2) and systemic primary carnitine deficiency (n = 1). The false positives were later confirmed to be carrier of citrullinemia type II (n = 2), carrier of glutaric acidemia type I (n = 1) and carrier of systemic primary carnitine deficiency (n = 1). There were no false negatives reported. The incorporation of a second-tier genetic screening test by NGS greatly enhanced our program's performance with 5-working days turn-around time maintained as before. In addition, early genetic information is available at the time of recall to facilitate better clinical management and genetic counseling.

Anesthetic managements, morbidities and mortalities in retroperitoneal sarcoma patients experiencing perioperative massive blood transfusion.

Given high risks of major bleeding during retroperitoneal sarcoma(RPS) surgeries, severe complications and deaths are common to see perioperatively. Thus, effective anesthetic management is the key point to ensuring the safety of patients. This study aimed to introduce anesthesia management and mortalities in RPS patients receiving massive blood transfusions during surgeries. Records of RPS surgeries under general anesthesia from January 2016 through December 2021 were retrospectively retrieved from our database. Patients who received massive blood transfusions (MBT) exceeding 20 units in 24h duration of operations were finally included in this study. Demographics, modalities of anesthesia management, blood loss, transfusion, peri-anesthesia biochemical tests as well as morbidities and mortalities were collected. Risk factors of postoperative 60d mortality were determined through logistic regression in uni-and multi-variety analysis using the statistics software STATA 17.0. A total of 70 patients (male 31) were included. The mean age was 50.1 ± 15.8 years. All patients received combined resections of sarcoma with involved organs under general anesthesia. Mean operation time and anesthesia time were 491.7 ± 131.1mins and 553.9 ± 132.6mins, respectively. The median intraoperative blood loss was 7000ml (IQR 5500,10000ml). Median red blood cells (RBC) and fresh frozen plasma (FFP) transfusion were 25.3u (IQR 20,28u), and 2400ml (IQR 2000,3000ml), respectively. Other blood products infusions included prothrombin complex concentrate (PCCs), fibrinogen concentrate (FC), platelet(plt) and albumin(alb) in 82.9% (58/70), 88.6% (62/70), 81.4% (57/70) and 12.9% (9/70) of patients. The postoperative severe complication rate(Clavien-Dindo grade≥3a) was 35.7%(25/70). A total of 7 patients (10%) died during the postoperative 60-day period. BMI, volumes of crystalloid infusion in anesthesia, and hemoglobin and lactate levels at the termination of operation were found significantly associated with postoperative occurrence of death in univariate analysis. In logistic multivariate analysis, extended anesthesia duration was found associated with postoperative venous thrombosis embolism (VTE) and severe complication. The lactate level at the immediate termination of the operation was the only risk factor related to perioperative death (p<0.05). RPS patients who endure MBT in surgeries face higher risks of death postoperatively, which needs precise and effective anesthesia management in high-volume RPS centers. Increased blood lactate levels might be predictors of postoperative deaths which should be noted.

Screening of Haliclona sp. symbiont bacteria that have the potential as MDR (Multidrug-Resistant) antibacterial from Tanjung Tiram Beach

Sponge Haliclona sp. is a type of sponge living in marine waters and is found in many areas of coral reef ecosystems. This study aimed to determine the potential for sponge Haliclona sp. obtained from Tanjung Tiram waters, Konawe, as a source of bioactive compounds of MDR (Multidrug-Resistant) antibacterial. The isolation method of Haliclona sp. symbiont was carried out by pouring and purification of Haliclona sp. symbiont using the scratch method. The purification results obtained 12 isolates of symbiont bacteria that were successfully isolated. Then the bacterial isolate was tested for its antibacterial ability against MRSA and ESBL Escherichia coli bacteria qualitatively using the agar diffusion method. Bacteria that have strong inhibitory activity are identified by morphological and biochemical tests. The results of the qualitative screening test produced four bacterial isolates that have inhibitory activity against MRSA (Methicilin Resistant Staphylococcus aureus) pathogenic bacteria with the isolate codes H3 and H8 belonging to the strong category while H4 and H10 belonging to the medium categoy. Additionally, two bacterial isolates that have inhibitory activity against ESBL Escherichia coli pathogenic bacteria with the isolate codes H3 and H8 belonging to the strong category. Based on morphological observations and biochemical tests, the H3 bacterial isolate was identified as the genus Corynebacterium spp. and the H8 bacterial isolate was identified as the genus Micrococcus spp. In conclusion, Haliclona sp. sponge exhibits promising potential as a source of microorganism producing antibacterial compounds, particulary againts MDR (Multidrug-Resistant) strains.

Treatment of tannery effluent: An effective biological enhancement approach

AbstractThe water pollution due to tannery industries that contaminate river water and other spheres, is a side effect of industry expansion. The research focused on the biological removal of synthetic azo dyes by using biological treatment under aerobic conditions. Through morphological observation, biochemical test, and 16S rRNA sequence analysis the isolated bacteria were determined to be Aeromonas caviae, Enterobacter sichuanensis. Reduction of the organic content present in the tannery effluent and removal of the nutrients that cause pollution or inactivate potential pathogenic microorganisms or parasites. Optimized the azo dye degradation conditions at the temperature of 37°C, at pH 7 in 10% inoculum concentration at the time of 48th hour where novel organism capable of performing effective degradation. The metabolite of an aliquot mixture of the optimized condition was analyzed using Fourier Transform Infrared (FTIR) and the results confirmed the reduction of azo dyes. These results indicate that A. caviae and E. sichuanensis can be utilized as a solution for the remediation of tannery industrial effluent containing azo dyes.

Antimicrobial resistance patterns, virulence genes, and biofilm formation in enterococci strains collected from different sources

BackgroundCurrently, antibiotic-resistant strains of Enterococcus are considered to be one of the critical health challenges globally. This study aimed to investigate the antibiotic susceptibility pattern, biofilm formation capacity, and virulence genes of enterococci isolated from different sources.MethodsIn this cross-sectional study, environmental and fecal samples were collected from the hospital environment, volunteers, and hospital staff from October 2018 to August 2019. The isolates were identified by morphological and biochemical tests (gram staining, catalase, bile resistance, esculin hydrolysis, carbohydrate fermentation, growth in 6.5% NaCl, Pyrrolidonyl arylamidase, arginine dehydrolase), and PCR for ddl gene. An antimicrobial susceptibility test was performed by the standard disk agar diffusion method according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Quantitative microplate assays were used to assess biofilm production. The bacterial DNAs were extracted by alkaline lysis method and polymerase chain reaction technique was used detect the esp, ace, and efaA virulence genes.ResultsOut of 145 isolates, 84 (57.9%) were identified as E. faecalis and 61 (42.1%) as E. faecium. Resistance to kanamycin and quinupristin-dalfopristin was 82.1% (69/84) and 85.7% (72/84), respectively, in E. faecalis isolates. Out of 61 E. faecalis isolates, 38 (62.4%) were resistant to kanamycin. Among the E. faecalis isolates, esp was the most dominant virulence gene (73.80%), followed by efaA, and ace, which were detected in 60.71%, and 30.95% isolates, respectively. In total, 68.27% of the strains were biofilm producers. Further, esp and efaA genes were more frequently found among E. faecalis strains with moderate and strong biofilm biomass.ConclusionsAccording to the findings of our study, enterococci strains isolated from different samples possess distinctive patterns of virulence genes. The esp, ace, and efaA genes were more prevalent among E. faecalis than E. faecium. Besides, the high level antibiotic resistance of normal flora and environmental enterococci strains is alarming the researchers.

The Influence of Lactobacillus spp. Secondary Metabolites Isolated from Immature Egyptian Honey on Human Pathogens, Transcription of Virulence Genes and Lung Cancer

The Influence of Lactobacillus spp. Secondary Metabolites Isolated from Immature Egyptian Honey on Human Pathogens, Transcription of Virulence Genes and Lung Cancer

Evaluation of Some Selected Medical Equipment in Olabisi Onabanjo University Teaching Hospital for Staphylococcus aureus of Nosocomial Threat

Hospital is an establishment where patients that are diseased, infected, ill or injured receive medical care. A medical staff and equipment are required for the task of medical care delivery. The equipment that are employed in hospital could be classified as critical, semi-critical and non-critical depending on their design and tasks. A total of Eighty-five (85) samples of hospital equipment swab were obtained from Olabisi Onabanjo Teaching Hospital of which 40 samples of Staphylococcus aureus were isolated. The isolates were Gram stained followed by conventional biochemical test for the identification of Staphylococcus aureus. Antibiogram of the isolates was determined. The isolates exhibited resistance to ampiclox (87.5%), zinnacef (92.5%), amoxicillin (92.5%), rocephin (80%), and septrin (55%), while susceptible to pefloxacin (77.5%), gentamicin (80%), streptomycin (72.5%), and ciprofloxacin (52.5%). Some of the resistant isolates were exposed to plasmid DNA analysis and were found to be plasmid borne of varied molecular weight, which could be responsible for resistance to the antibiotics observed. There is therefore a need for regular disinfection, properly sterilization and preservation of medical equipment before and after use, which could curtailed or reduce the spread of equipment borne nosocomial infection.

Retrospective evaluation of leflunomide as an adjunctive therapy in dogs with non-associative immune-mediated thrombocytopenia: 20 cases (2008-2021).

To describe leflunomide as an adjunctive therapy in the treatment of non-associative immune-mediated thrombocytopenia. A retrospective study of dogs with a diagnosis of non-associative immune-mediated thrombocytopenia treated with leflunomide March 2008 to September 2021 was conducted. Data collected included signalment, clinical signs, physical examination findings and diagnostic testing performed. Medications administered, duration of hospital stay, time to platelet concentration >150×109/L and adverse events during leflunomide therapy were recorded. Relapses within a year of diagnosis were reported. A total of 20 client-owned dogs met inclusion criteria. Nineteen of 20 dogs (95%) achieved a platelet concentration >150×109/L with leflunomide and prednisone combination therapy and four dogs (21.1%) relapsed during treatment or shortly after treatment. Adverse effects included diarrhoea (n=5), mild lymphopenia (n=9) and mild intermittent anaemia (n=1). A single dog developed hepatotoxicity presumed to be secondary to leflunomide therapy that resolved after drug discontinuation. One dog was treated for aspiration pneumonia during treatment. Two dogs were euthanased while receiving leflunomide. Length of hospitalisation, time to platelet recovery, treatment response and relapse rate were comparable with alternative treatment protocols. Most adverse effects did not require leflunomide dose adjustment; however, two dogs died while undergoing leflunomide treatment and there is compelling evidence that one of these dogs experienced fatal infection secondary to immune-suppression. Hepatotoxicity remains a known complication of leflunomide treatment and serial biochemistry testing is recommended.

Optimizing d-mannose and glyceraldehyde concentrations as glucose preservatives without clinically affecting biochemical test results

Objectives: The objectives were to evaluate blood additives that combined lithium heparin (LH)-salt with glyceraldehyde (GLY) or d-mannose (MAN) for preserving glucose levels in plasma samples and to simultaneously determine the compatibility of these additives with 14 other biochemical tests. MethodsBlood samples from 40 subjects, equally divided into healthy and diabetic groups, were collected using five different additives. The three most effective additives, LH/GLY, LH/MAN, and LH/GLY/MAN, were selected for ensuring the best preservation of glucose levels and compatibility with 14 biochemical tests. One-way analysis of variance was used to analyze the mean paired differences of glucose level and biochemical tests. Simultaneously, the clinical criteria from Johns Hopkins Hospital were used to guide the interpretation and set acceptable thresholds for measurements that exceeded the standards. ResultsThe combination of 160 mmol/L GLY, 8.4 mmol/L MAN, and LH, maintained glucose levels at approximately 93.4–93.7 % for healthy subjects and 91.3–92.8% for subjects with diabetes mellitus over 8 h. The mean paired differences of glucose levels in preservation were statistically insignificant. The biases in 14 biochemical tests for LH/GLY/MAN and LH/MAN remained within the acceptable clinical criteria during the 8 h. ConclusionsCombining 160 mmol/L GLY, 8.4 mmol/L MAN, and LH, proved more effective in maintaining glucose levels than individual additives or the conventional sodium fluoride preservative. It did not yield clinical discrepancies in the 14 biochemical tests during 8 h at room temperature.

Seed vigour testing of Acacia mangium Willd. as a result of mutation

Plant breeding techniques are applied to increase the genetic variation of a plant. Induction of plant mutation using gamma irradiation is one of the methods that can be applied to improve the genetic diversity of plants. This study aims to analyze the vigour of mangium seeds treated by mutation. Mutation treatment was administered to M0 of mangium seed. This study used M2 mangium seeds and control mangium seeds (without mutation treatment). Several assays, such as growth, stress, and biochemical tests, were conducted to determine the mangium seed’s vigour. The growth test showed that M2 mangium seeds had higher germination values, germination height, dry weight, and vigour index than control mangium seeds. The stress test consisting of the Hiltner test, accelerated ageing test, and saturation test of the germination value of M2 mangium seeds wass higher than control mangium seeds. In the biochemical test, the electrical conductivity value of control mangium seeds was more significant than M2 mangium seeds. In general, M2 mangium seeds had higher vigour than control mangium seeds.

Saikosaponin D mitigate pilocarpine-induced astrocyte injury by regulating the NLRP3/caspase-1 signaling pathway.

Studies have shown that saikosaponin D (SSD) has favorable neurotherapeutic effects. Therefore, the objective of this study was to explore the efficacy and possible molecular mechanisms of SSD on pilocarpine (PP)-induced astrocyte injury. Primary astrocytes were isolated from juvenile rats and identified using immunofluorescence. The cells were treated with PP and/or SSD for 6 h and 12 h, respectively, followed by measurement of their viability through 3-(4,5-dimethylthiazol)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Next, quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the expression levels of Glial fibrillary acidic protein (GFAP), C3, S100 calcium binding protein A10 (S100a10), pentraxin 3 (Ptx3), toll-like receptor 4 (TLR4), and RAG in astrocytes after different treatments. Enzyme-linked immunosorbent assay and biochemical tests were utilized to evaluate the level of inflammatory factors [interleukin (IL)-1β, IL-6, and tumor necrosis factor alpha (TNF-α)] secreted by cells and the content of oxidative stress-related factors (malondialdehyde [MDA] and glutathione [GSH]) or enzyme activity (catalase [CAT] and glutathione peroxidase [GPX]) in cells. The JC-1 mitochondrial membrane potential (MMP) fluorescence probe was used to measure the MMP in astrocytes. Additionally, western blot was applied to test the expression of proteins related to the nod-like receptor protein 3 (NLRP3)/caspase-1 signaling pathway. PP treatment (1 mM) induced cell injury by significantly reducing the viability of astrocytes and expression of cellular markers. SSD treatment (4 μM) had no toxicity to astrocytes. Besides, SSD (4 μM) treatment could significantly up-regulate the cell viability and marker expression of PP-induced astrocytes. Furthermore, SSD could be employed to inhibit inflammation (reduce IL-1β, IL-6, and TNF-α levels) and oxidative stress (decrease MDA level, elevate GSH level, the activity of CAT and GPX), and ameliorate mitochondrial dysfunction (upregulate JC-1 ratio) in PP-induced astrocytes. Moreover, further mechanism exploration revealed that SSD treatment significantly reduced the activity of the NLRP3/caspase-1 signaling pathway activated by PP induction. SSD increased cell viability, inhibited inflammation and oxidative stress response, and ameliorated mitochondrial dysfunction in PP-induced astrocyte injury model, thus playing a neuroprotective role. The mechanism of SSD may be related to the inhibition of the NLRP3/caspase-1 inflammasome.

Parenteral vs Enteral Nutrition: A Retrospective Study on PNI and Inflammatory Status

Background: Malnutrition consists of a functional and structural alteration caused by an imbalance between specific needs, intake, or utilization of foods and nutrients. Objective: The aim of this trial is to establish whether PNI (Prognostic Nutritional Index) is related to the inflammation pattern in our patients and which type of AN gives more advantages in clinical practice in improving PNI, and so the prognosis. Methods: During hospitalization, we have identified the presence of malnutrition and its aetiology- type, and initiated artificial nutrition (AN), enteral (EN), or parenteral (PN). For efficacy, we can use many biochemical tests, such as pre-albumin, transferrin, RBP, PINI, and albumin. The retrospective study was conducted by analyzing 145 patients. At baseline and after 7 days, the concentration of Albumin, Platelets, Neutrophils, Lymphocytes, and RCP was measured and was calculated PNI, PLR, and NLR. The patients were divided into three groups according to the treatment received with artificial nutrition: 32 Parenteral (PG), 15 Enteral (EG), and 17 Combined (CG). Results: Between baseline and 7 days, no change was observed, while in EG a significant increase in albumin and RCP and a significant reduction in PNI were observed, and, in CG a significant increase in lymphocytes, in PNI and a significant reduction in NLR. Conclusion: In conclusion, a short-term nutritional intervention can benefit from a combined approach to improve prognostic such as PNI.

Pattern of antibiotic resistance in surgical site infections in a tertiary care hospital of Nepal

Background: Surgical site infection (SSI) is the second most common hospital-acquired infection. The identification of bacterial pathogens and their antibiotic susceptibility pattern is required for the successful treatment of SSI and curb antimicrobial resistance, which is a major threat globally. This study aimed to assess the microbial profile of the organisms causing SSI and their antibiogram in a tertiary care hospital. Methods: This cross-sectional study was conducted at Microbiology Laboratory of tertiary care hospital, from March 2023 to August 2023. Swab or aspirate specimens were collected aseptically from the surgical site and processed for bacterial isolation following standard bacteriological techniques. Gram’s staining and biochemical tests were performed to identify the organisms at the species level. Antimicrobial susceptibility tests were done following Kirby–Bauer’s disc diffusion method. Statistical analysis was performed using SPSS software version16.0. Results: Out of 405 samples, 92 (22.7%) yielded bacterial growth. Eighty-three culture positive cases (90.2%) were monomicrobial while nine (15.7%) had mixed growth of at least two different bacteria. Out of 108 isolates, 43 (39.8%) were gram-positive and 65 (60.2%) were gram-negative bacteria. Escherichia coli (39.8%) was the most commonly isolated organism. All gram-negative isolates were resistant to ceftriaxone followed by amoxicillin-clavulanic acid (94.0%), amoxicillin (94.0%), cefixime (90.7%), and cefepime (89.8%). Similarly, none of the gram-positive isolates were susceptible to ampicillin, amoxicillin, and ampicillin/sulbactam. The most effective drugs against E. coli were imipenem (100%), followed by tigecycline (96%), meropenem (95.1%), chloramphenicol (84%), amikacin (81.5%), and gentamicin (81.5%). Overall prevalence of MDR was 59.3%. Conclusion: Our study demonstrates increased resistance of both gram-positive and gram-negative organisms to commonly used drugs like cephalosporins and even quinolones. Continued surveillance of antimicrobial susceptibility results at local level, dissemination of data, and prescribing the antibiotics accordingly based on culture and sensitivity results are necessary to ameliorate antimicrobial resistance.

Yanggan Jiangmei Formula alleviates hepatic inflammation and lipid accumulation in non-alcoholic steatohepatitis by inhibiting the NF-κB/NLRP3 signaling pathway

The role of NF-κB and the NLRP3 inflammasome in the chronic inflammatory microenvironment of non-alcoholic steatohepatitis (NASH) has been posited as crucial. The Yanggan Jiangmei Formula (YGJMF) has shown promise in ameliorating hepatic steatosis in NASH patients, yet its pharmacological mechanisms remain largely unexplored. This study was conducted to investigate the efficacy of YGJMF in NASH and to elucidate its pharmacological underpinnings. To simulate NASH both in vivo and in vitro, high-fat-diet (HFD) rats and HepG2 cells stimulated with free fatty acids (FFAs) were utilized. The severity of liver injury and lipid deposition was assessed using serum indicators, histopathological staining, micro-magnetic resonance imaging (MRI), and the liver-to-muscle signal intensity ratio (SIRL/M). Furthermore, a combination of enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), immunofluorescence, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blotting analyses was employed to investigate the NF-κB/NLRP3 signaling pathway and associated cytokine levels. The results from liver pathology, MRI assessments, and biochemical tests in rat models demonstrated YGJMF's significant effectiveness in reducing liver damage and lipid accumulation. Additionally, YGJMF markedly reduced hepatocyte inflammation by downregulating inflammatory cytokines in both liver tissue and serum. Furthermore, YGJMF was found to disrupt NF-κB activation, consequently inhibiting the assembly of the NLRP3 inflammasome in both the in vitro and in vivo models. The preliminary findings of this study suggest that YGJMF may alleviate hepatic steatosis and inhibit the NF-κB/NLRP3 signaling pathway, thereby exerting anti-inflammatory effects in NASH.

Detection of Novel spa Types in Clinical Isolates of Methicillin-Resistant Staphylococcus aureus in Iraq

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a gram-positive bacterium that is an opportunistic pathogen, causing infections in hospital settings and communities. MRSA has become a significant and increasing problem in Iraq. The aim of this study was to evaluate the genetic mutations of MRSA strains, especially in the spa gene, isolated from patients in Wassit, Iraq.Methods and Results: Biochemical tests were conducted to identify S. aureus isolates and then on MRSA. The MRSA was identified by the Chrome Agar method and confirmed by PCR with genotyping of the mecA gene. The disk diffusion method was used to detect antibiotic resistance to three different common antibiotics used at Wassit hospitals. The Vitek-2 compact system was utilized for the detection of the minimum inhibitory concentration (MIC) of vancomycin. All MRSA strains in this study were tested to screen the mecA gene, with 21 strains subjected to the molecular typing method for the spa gene. Out of 166 samples, 132(79.5%) contained S. aureus and 34(20.4%) were identified as MRSA. Genotyping showed that out of 34 MRSA, 31(91.2%) isolates were mecA-positive. The spa gene was detected in 21(61.8%) isolates out of 34 MRSA samples. The spa typing of 21 MRSA samples revealed four different spa types, as follows: t386 (3/14.3%), t3576 (1/4.8%), t10002 (1/4.8%), and t10234 (1/4.8%). High polymorphism rates were shown in isolates of spa type t386. Conclusion: Our data represent the first report to detect novel mutations in the spa gene in the MRSA clinical isolates from Wassit hospitals, Iraq.