Urine Samples Of Patients Research Articles

ISOLATION AND CHARACTERIZATION OF ESCHERICHIA COLI IN UTI PATIENTS OF DISTRICT D.I.KHAN AND TO PROBE THEIR ANTIBIOTIC RESISTANCE

Background: Globally urinary tract infections are present predominantly and mostly UTIs are present or noticed in the developing countries like Pakistan, Bangladesh etc. There are multiple factors in spread of UTIs like microbes, taking certain medicines, sexual activities, poor hygiene, not taking enough water etc. Objectives: The prevalence of UTIs and development of resistance against the antibiotics is due to unjustified use of antibiotics. Methodology: We performed a study in a remount area of Khyber Pakhtunkhwa i,e Dera Ismail Khan. We conducted a research study consist of a three months duration. Hundred (100) urine samples were collected from the patients who visited District Head Quarter (DHQ) teaching hospital Dera Ismail Khan. Patients having different age groups were included in study, Results: After processing samples, the number of samples having E.coli i,e positive samples were 73 in number, which were confirmed on the basis of different tests. The prevalence of UTIs due to E. coli organism was noticed increased in female patients (69 %) as compared to male patients and its prevalence was 31%.When the isolated E. coli strain were tested against different antibiotics, showed that those strains which were isolated from urine samples of the male patients of age group 61 to 80 years showed higher sensitivity against PTZ, NF, FOS, AK, IMP, and SCF, while in female high rate of sensitivity against PTZ, NF, FOS, AK, IMP, and SCF antibiotics was observed against E. coli strains which were isolated from the patients having age from 21 to 40 years Likewise E.coli isolates were sensitive against Imipenem, Peracillin + Tazobactam, Cefoperazone + sulbactam, Fosfomycin, Amikacin, Nitrofurantoin, Peracillin and Tambactam irrespective of age of the patients from which bacteria were isolated. Conclusion: Furthermore, present study also revealed, that the E. coli has developed resistance against commonly used antibiotics which is a serious concern to worry about and serious steps should be taken by the authorities to stop frequent use of antibiotics without proper prescription to avoid this issue.

Urine PLA2R Antibody Detection in Hazard Stratification of PLA2R-Associated Membranous Nephropathy.

M-type phospholipase A2 receptor (PLA2R) is the major autoantigen of membranous nephropathy (MN). As the specific antibodies of MN, the correlation between serum PLA2R antibody (sPLA2R-Ab) levels and PLA2R-associated MN (PMN) risk stratification is still controversial. To apply the time-resolved fluorescence immunoassay (TRFIA) method on urine PLA2R-Ab (uPLA2R-Ab), detect, and then establish a more sensitive method of combined serum and urine PLA2R-Ab detection for PMN hazard stratification. A highly sensitive TRFIA method was used to detect the initial serum and urine samples of patients with PMN. Patients were grouped into remission and nonremission groups according to the outcomes after 12 months of treatment and the data were analyzed. The cutoff values of sPLA2R-IgG (sPLA2R-immunoglobulin G), uPLA2R-IgG, sPLA2R-IgG4, and uPLA2R-IgG4 for distinguishing between remission and nonremission groups were 50 relative units (RU)/mL, 3.51 RU/mL, 6835 ng/mL, and 143.4 ng/mL, respectively. The average value in the remission group for sPLA2R-IgG, uPLA2R-IgG, sPLA2R-IgG4, and uPLA2R-IgG4 was 37.39 RU/mL, 1.10 RU/mL, 3498.99 ng/mL, and 33.83 ng/mL, respectively. The average value in the nonremission group for sPLA2R-IgG, uPLA2R-IgG, sPLA2R-IgG4, and uPLA2R-IgG4 was 279.96 RU/mL, 45.36 RU/mL, 25762.47 ng/mL, and 1383.89 ng/mL, respectively. For sPLA2R-Ab as the primary factor, in combination with uPLA2R-Ab, the high-risk predictive value of combined detection of serum and urine PLA2R-IgG and of serum and urine PLA2R-IgG4 was upgraded from 54.55% to 100% and from 75% to 100%, respectively. A highly sensitive TRFIA method was applied in this study; the combined detection of serum and urine PLA2R-Ab improves the efficiency of PMN risk stratification, and can provide a better assessment of PMN monitoring.

Non-invasive diagnosis of papillary thyroid microcarcinoma using a novel metabolomics analysis of urine.

The severity of thyroid cancer is judged on the basis of histologic and clinical features. A limited number of studies have considered urinary metabolite signatures for its diagnosis, and no reliable urinary metabolite biomarkers have been proposed. This diagnostic method would be particularly valuable because of its non-invasive nature. A nuclear magnetic resonance (NMR)-based metabolomics approach was used as the analytical platform to study the urine samples of patients with PTMC. Urine samples collected from 41 PTMC patients, 52 healthy subjects, and 13 patients with benign tumors were analyzed using 1H-NMR spectroscopy to identify metabolic changes. PLS-DA, or partial least squares discriminant analysis, was used to analyze the NMR spectra. A double cross-validation method and randomization tests were used to validate PLS-DA models. Clear discriminations between PTMC patients and healthy controls, as well as between PTMC patients and patients with benign tumors were obtained. Collectively, pi-methyhistidine, trimethylamine, myo-inositol, acetate, suberate, azelate, mannitol, tau-methylhistine, ascorbate, 3-aminoisobutyric acid, 2-oxoglutarate, and methanol contributed to the discrimination. Apart from myo-inositol and methanol, all of these metabolites exhibited increased levels in the urine samples of PTMC patients as compared to that of patients with benign tumors. The application of this NMR-based metabolomics approach allowed the detection of anomalous metabolic traits directly connected PTMC, potentially yielding a more sensitive and comprehensive diagnostic results for PTMC.

Microparticle-assisted protein capture method facilitates proteomic and glycoproteomic analysis of urine samples

Serum tests have become a partial alternative to renal biopsy for diagnosing primary membranous nephropathy (pMN). However, urine tests, due to their non-invasive nature and ability to more accurately reflect glomerular diseases, hold great promise for the detection of pMN. However, the low protein concentration and the time-consuming sample preparation procedure of urine samples challenges the proteomic and glycoproteomic analysis to find urine-derived signatures associated with pMN. In this study, we presented a microparticle-assisted protein capture (MAPC) method to efficiently prepare urine samples. It was found that proteins and N-linked intact glycopeptides can be sensitively identified from urine samples of pMN patients and healthy controls by using this method. For comparison, proteins and N-linked intact glycopeptides from serum were also analyzed. Interestingly, discrepancy was found in the changing trends for proteins/intact glycopeptides between serum and urine. Moreover, urine derived proteins, N-linked intact glycopeptides, and glycans exhibited more pronounced changes between pMN and healthy control compared to serum sample. Notably, urine IgG4 not only up-regulated in pMN at global peptide level, its corresponding site-specific glycans were specifically identified in pMN urine with significantly up-regulations, suggested the potential of using glycosylated urine IgG4 for the non-invasive diagnosis of pMN.

Urine and serum-based ELISA using a recombinant protein and synthetic peptide for the diagnosis of tegumentary leishmaniasis

The diagnosis of tegumentary leishmaniasis (TL) presents problems by the variable sensitivity and specificity of the tests, and the biological samples used are also invasive. Here, ELISA experiments were performed using paired TL patient urine and serum samples in reaction against the recombinant LiHyS protein, a predicted B cell epitope and parasite antigenic extract (SLA). Two hundred and five paired samples were used, which were provided by TL patients, healthy controls and patients with Chagas disease, leprosy, malaria or HIV-infected. An urine-based ELISA showed sensitivity values of 100%, 92.1%, and 82.5%, when rLiHyS, peptide and SLA were used, respectively; and specificity of 100%, 87.6%, and 79.5%, respectively. A serum-based ELISA showed sensitivity values of 100%, 99.3%, and 81.5%, using rLiHyS, peptide and SLA, respectively, and sensitivity of 100%, 96.5%, and 72.2%, respectively. In both cases, rLiHyS presented the better performance to diagnose TL by using patient serum and urine.

Molecular detection of Staphylococcus aureus in urine in patients with S. aureus bacteremia: an exploratory study.

Staphylococcus aureus bacteremia (SAB) is associated with a 90-day mortality of 28-34%. Many SAB-patients (7.8-39%) have a secondary S. aureus bacteriuria (SABU) mainly without symptoms of a urinary tract infection. Due to high morbidity and mortality, there is an interest in rapid detection of S. aureus bacteremia. Here, we compared a rapid nucleic acid amplification test (NAAT) with conventional culture to detect S. aureus in urine and to identify cases with increased risk for SAB. In a cross-sectional study, we assessed urine samples (mid-stream, clean catch and catheter urine) of patients with SAB and bacteremia other than SAB (non-SAB). Urine samples were collected ± 3 days to the collection of the positive blood culture and were cultured on a set of selective and non-selective agar plates. NAAT was performed using a commercial test (Xpert® SA Nasal Complete G3, Cepheid) from a sterile swab soaked in urine. We included samples from 100 patients (68% male, median age: 67.4 years) with SAB and 20 patients (75% male, median age: 65.84 years) with non-SAB. The sensitivity of detecting SAB from urine samples was 47% (specificity: 90%) for NAAT, when applying a Ct-value of ≤ 37.4 for positive results. Urine culture had a sensitivity of 25% and a specificity of 95%. Molecular and culture methods showed a moderate agreement (80%, Cohens kappa: 0.55). NAAT from urine has a higher sensitivity than culture in patients with SAB and could potentially identify cases with increased risk for SAB. Future studies should investigate whether this characteristic could translate into a clinical benefit through rapid detection of SAB.

In Search of Relevant Urinary Biomarkers for Thyroid Papillary Carcinoma and Benign Thyroid Nodule Differentiation, Targeting Metabolic Profiles and Pathways via UHPLC-QTOF-ESI+-MS Analysis.

Identification of specific urine metabolic profiles for patients diagnosed with papillary thyroid carcinoma (TC) vs. benign nodules (B) to identify specific biomarkers and altered pathways compared to those of healthy controls (C). Patient urine samples were collected, before surgery and after a histological confirmation of TC (n = 30) and B (n = 30), in parallel with sample collection from healthy controls (n = 20). The untargeted and semi-targeted metabolomic protocols were applied using UPLC-QTOF-ESI+-MS analysis, and the statistical analysis was performed using the Metaboanalyst 6.0 platform. The results for the blood biomarkers, previously published, were compared with the data obtained from urine sampling using the Venny algorithm and multivariate statistics. Partial least squares discrimination, including VIP values, random forest graphs, and heatmaps (p < 0.05), together with biomarker analysis (AUROC ranking) and pathway analysis, suggested a specific model for the urinary metabolic profile and pathway alterations in TC and B vs. C, based on 190 identified metabolites in urine that were compared with the serum metabolites. By semi-targeted metabolomics, 10 classes of metabolites, considered putative biomarkers, were found to be responsible for specific alterations in the metabolic pathways, from polar molecules to lipids. Specific biomarkers for discrimination were identified in each class of metabolites that were either upregulated or downregulated when compared to those of the controls. The lipidomic window was the most relevant for identifying biomarkers related to thyroid cancer and benign conditions, since this study detected a stronger involvement of lipids and selenium-related molecules for metabolic discrimination.

Investigation of Lead, Nickel, and Cadmium Levels in Urine Samples of Healthy and Patients with Kidney Failure in Al-muthanna Governorate, Iraq

Background: The Iraqi ecosystem, particularly in the southern area, has been polluted due to human activity. Analyzing biological materials is the most common method for detecting the presence of toxic substances in the human body. Method: Heavy metal levels of Pb, Ni, and Cd in urine specimens collected from individuals with renal failure and healthy individuals residing in Al-Muthanna province were measured by using atomic absorption spectroscopy. Urine Specimens were collected from two cohorts of male and female participants: the group of individuals with renal failure and the group of individuals who are in good condition. The specimens of urine from both the group of patients with renal failure and the group of healthy individuals were taken from the Al-Muthanna governorate in southern Iraq. This governorate served as a focal point for extensive military operations throughout the Gulf War. Results: The concentrations of toxic substances (Pb, Ni, and Cd) in the urine specimens of the cohort of patients suffering from renal failure are 0.411, 0.197, and 0.113 mg/l, respectively. Concentrations of (Pb, Ni, and Cd) in the healthy group are 0.249, 0.101, and 0.0294 mg/l, respectively. The toxic metals found in urine samples can be organized in the following order: Pb &gt; Ni &gt; Cd. Conclusion: The findings indicated that the concentrations of toxic substances in specimens of urine from individuals with renal failure are considerably greater than those in the control group of healthy individuals. According to the findings, patients' incidence of renal failure may be related to the prevalence of harmful compounds in southern Iraq.

Prevalence and virulence profiles of ESBL-producing Escherichia coli in urinary and blood infections in South Korea.

Escherichia coli is a significant pathogen in extraintestinal infections, and ESBL-producing E. coli poses a major clinical challenge due to its antibiotic resistance. This study comprehensively analyzed E. coli isolates from urine and blood samples of patients with urinary tract and bloodstream infections at three major tertiary hospitals in South Korea. The goal was to provide insights into the distribution, antibiotic resistance, and virulence factors of these strains. Our analysis identified CTX-M and TEM as the dominant ESBL types, found in 71.7% and 61.7% of isolates, respectively, with 46.7% showing co-occurrence. Multilocus sequence typing (MLST) revealed the predominance of high-risk clones such as ST131, ST69, ST73, and ST95, with rare sequence types like ST410 and ST405 also identified. The high prevalence of virulence factors, including iutA (80.8%) and kpsMII (74.2%), further highlights the complexity of these strains. In addition, 38.3% of clinical isolates contained a combination of siderophore, adhesin, protectin, and toxin-related genes. There was no significant difference between urinary tract and bloodstream infections or regional differentiation in Korea. This study highlights the importance of controlling ESBL-producing E. coli infections, especially given the increasing incidence among patients with underlying medical conditions and older adults who are more susceptible to urinary tract infections. These findings serve as valuable indicators for pathogen analysis, especially those harboring antibiotic resistance and toxin genes. The insights gained are expected to contribute significantly to the development of infectious disease prevention and control strategies.

Detection of renal tubular transporter proteins in canine urinary extracellular vesicles using liquid chromatography-tandem mass spectrometry.

Urinary extracellular vesicles (UEVs) are membranous particles that carry renal tubular transporter proteins. Here, we evaluate whether selected renal tubular transporter proteins can be detected in UEVs isolated from small volume (1-5 mL) canine urine samples of healthy dogs and canine patients with elevated circulating parathyroid hormone (PTH)/PTH-related peptide (PTHrp) concentrations, hypercortisolism, and primary hypoadrenocorticism using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The total creatinine content of each urine sample was calculated from urine volume and creatinine concentration. UEVs were isolated by size exclusion chromatography prior to quantification by nanoparticle tracking analysis and proteomic analysis by LC-MS/MS. Group comparisons were made using non-parametric statistics. Aquaporin-2 (AQP2) and the renal sodium/phosphate co-transporter (NPT2A) were detected in UEVs isolated from small volume samples of almost all healthy dogs but were not detected in most dogs with elevated circulating PTH/PTH related peptide (PTHrp) concentrations, hypercortisolism and primary hypoadrenocorticism. Total creatinine content of the urine sample was strongly positively correlated with the number of UEVs (rs = .84, P < .001); thus, total creatinine was used as a surrogate marker of UEV number. In healthy dogs, AQP2 and NPT2A were both detected in samples containing at least 1.7 × 109 UEVs or 24 μmol creatinine, however in non-healthy dogs, AQP2 and NPT2A were not detected in most samples containing up to 6.3 × 109 UEVs or 32 μmol creatinine.

Evaluation of a Point of Care Urine Drug Test System for Application in Outpatient Pain Management Clinics

Abstract Introduction/Objective Most states require testing for illicit and controlled substances in ambulatory, non-terminal pain management populations to ensure prescription compliance. Point of care (POC) urine drug testing (UDT) is a widely adopted testing method that assists with clinical decision making within minutes. However, these immunoassays may lack the sensitivity and specificity needed for potent medications such as fentanyl. This study compares the performance characteristics of a POC UDT to an in-house, high throughput, automated chemistry analyzer and the reference method, gas chromatography-mass spectrometry (GC-MS). Providers should be aware of the limitations of each methodology given the rise of synthetic opioid misuse, and the need to quantify multiple substances and metabolites within the same drug class. Methods/Case Report 100 residual deidentified patient urine samples from an ambulatory pain management population were analyzed using GC-MS, Profile®-V MEDTOX Scan® POCT device (Burlington, NC), and a Beckman Coulter AU480® Chemistry Analyzer using EMIT® II Plus reagents (Beckman Coulter; Brea, CA). Drug classes on the Profile®-V included: amphetamines, barbiturates, benzodiazepines, buprenorphine, cocaine metabolite, methamphetamine, methadone, opiates, oxycodone, cannabinoids and phencyclidine. Drug classes on the AU480® included the same except without a methamphetamine-specific immunoassay. Results (if a Case Study enter NA) The results from GC-MS showed that 99% (99/100) of the samples were positive for one or more of the drug classes. Overall, the AU480® was 90% sensitive and 98% specific while the MEDTOX Scan® was 87% sensitive and 98% specific. Both devices exhibited variable performance across drug classes. Neither the AU480® nor the MEDTOX Scan® detected fentanyl in five specimens. Conclusion POCT-UDS systems like the Profile®-V MEDTOX Scan® can provide good accuracy and may offer a time and cost benefit when compared to higher throughput analyzers like the AU-480®. Due to the low reference values of immunoassays, definitive testing such as GC-MS is recommended for use in the pain management population.

Interference Between S. Aureus and P. Aeruginosa Clinical Isolates

General Background: Understanding microbial interactions between Pseudomonas aeruginosa and Staphylococcus aureus is crucial for clinical infections, as they often coexist and influence each other's growth and virulence. Specific Background: Both organisms are known for their ability to form biofilms and exhibit multidrug resistance (MDR), complicating treatment strategies in hospitalized patients. Knowledge Gap: Previous studies have explored the virulence of P. aeruginosa and S. aureus, but there is limited understanding of their direct in vitro interactions, particularly in protease and lipase production. Aims: The study examined the interaction between P. aeruginosa and S. aureus isolates in hospitalized patients' sputum, urine, and blood samples, focusing on virulence factors, biofilm formation, and antibiotic resistance patterns. Results: In Diyala, Iraq, 50 clinical isolates showed P. aeruginosa as protease producers, lipase producers, and biofilm producers, with significant MDR phenotypes in both species. Novelty: This study highlights the ability of P. aeruginosa to produce staphylolysin, offering novel insights into the antagonistic mechanisms that may suppress S. aureus in co-infections. Implications: The study emphasizes the significance of understanding microbial interactions in clinical infections, particularly in biofilm-associated MDR infections, to improve treatment outcomes and guide more effective therapeutic approaches. Highlights: P. aeruginosa staphylolysin inhibits S. aureus growth in vitro. Both bacteria produce biofilms and exhibit multidrug resistance. Microbial interactions impact infection severity and treatment strategies. Keywords: Microbial interactions, Pseudomonas aeruginosa, Staphylococcus aureus, Biofilm, Multidrug resistance

Metabolomic profiling reveals the step-wise alteration of bile acid metabolism in patients with diabetic kidney disease

BackgroundDiabetic kidney disease (DKD) is the major complication of diabetes concomitant with gut dysbiosis and glycometabolic disorder, which are strongly associated with bile acid (BA) metabolism. Yet studies investigating the BA metabolism involving in DKD pathogenesis are limited. This study aimed to explore the metabolomic profiling of BAs in DKD and analyze its association with DKD progression.MethodsAn ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was established to quantify BAs in the plasma, fecal and urine samples of patients with DKD or T2DM and healthy individuals (n = 30 for each group). The key BAs associated with DKD were identified by orthogonal partial least-squares discriminant analysis (OPLS-DA) and receiver-operating characteristic (ROC) curve. Polynomial regression and Pearson’s correlation analyses were performed to assess the correlation between the key BAs and the clinical indicators reflecting DKD progression.ResultsMetabolomic profiling of 50 kinds of BAs presented the markedly step-wise alterations of BAs in plasma and feces as well as the little in urine of patients with DKD. Eight kinds of BAs in the plasma, eight kinds in the feces and three kinds in the urine were abnormally expressed, accompanying with the increased conjugated/unconjugated ratios of cholic acid, deoxycholic acid, chenodeoxycholic acid, ursodeoxycholic acid and hyocholic acid in the plasma, and of cholic acid, chenodeoxycholic acid and lithocholic acid in the feces. Moreover, the increased plasma level of glycochenodeoxycholic acid, and the increased fecal levels of glycolithocholic acid, 7-ketodeoxycholic acid and chenodeoxycholic acid-3-β-D-glucuronide are strongly correlated with the clinical indicators reflecting DKD progression, including eGFR, 24 h urinary protein and 24 h urinary microalbumin.ConclusionsOur study for the first time disclosed the specific alterations of BA metabolism reflecting the step-wise progression of DKD, providing the basis for early identification and therapeutical strategies for DKD.

Isolating and Identification Of Bacteria Causing Urinary Tract Infections From Hospital Patients And Nursing Staff

The current study aimed to isolate and identification the bacteria that cause urinary tract infections from recumbent patients and nursing staff, using traditional and molecular methods. 700 Clinical samples were collected from Azadi Teaching Hospital and Kirkuk Teaching Hospital in Kirkuk city for the period from October 2023 to March 2024 from (500) recumbent patients and 200 nursing staff. 347 (49.6%) of the total samples showed positive results for bacterial growth when cultured on ideal culture media, including mannitol, blood, and MacConkey agars. 353(50.4%) of total samples were appear as negative results for bacterial growth. The findings showed the distribution of bacterial isolates obtained from urine samples of recumbent patients and nursing staff at Azadi Teaching Hospital. Escherichia coli (44.6% and 41.67%) was the most prevalent bacterial species, followed by (13.08% and 13.89%) K. pneumoniae, (11.54% and 11.11%) P. aeruginosa. while, the distribution of bacterial isolates obtained from urine samples of recumbent patients and nursing staff at Kirkuk Teaching Hospital. Escherichia coli (46.9% and 51.85%) was the predominant bacterial species, followed by (16.6% and 14.81%) K. pneumoniae and (9% and 11.11%) P. aeruginosa. for molecular study, 7(50.0%) out of a total of 14 E. coli isolates possessed the gyrA gene. On the other hand, 12(100.0%) isolates out of a total of 14 isolates of K. pneumoniae possess the gyrA gene, while 11(91.7%) isolate out of a total of 12 isolates of P. aeruginosa possesses the gyrA gene. It is concluded that the most common cause of urinary tract infections is Escherichia coli followed by K. pneumoniae and P. aeruginosa, and that one of the most important causes of antibiotic resistance is the presence of the gyrA gene, which causes bacterial resistance to fluoroquinolone.

B-281 Performance Characteristics Evaluation of the ARK Diagnostics Immunoassay for Xylazine/4-HydroxyXylazine in Urine

Abstract Background Xylazine exposure is currently common in some US cities, but routine laboratory testing is not available. We evaluated the ARK Diagnostics immunoassay for qualitative detection of xylazine/4-hydroxy Xylazine in urine, prior to its anticipated release as a Forensic Use Only assay. Evaluation of performance characteristics of the ARK assay were conducted at two university hospitals, using either the semi-quantitative application of the assay, as performed on the Roche Cobas 503 analyzer (Institution A), or the qualitative application of the assay, as performed on the Beckman Coulter AU480 analyzer (Institution B). Methods Xylazine (X) and 4-hydroxy Xylazine (4-OHX) were obtained from Cayman Chemical (Ann Arbor, MI). Pooled xylazine-free urine was spiked either with X or 4-OHX in the range of 0-2000 ng/mL. De-identified, to-be-discarded patient urine samples were obtained from among those submitted for routine drugs-of-abuse panel testing. X measurements on all samples were performed by LC-MS/MS; samples were segregated into X-NEGATIVE (X&amp;lt;10 ng/mL) and X-POSITIVE (X&amp;gt;=10 ng/mL). The ARK assay was conducted according to manufacturer’s instructions, either as a semi-quantitative assay (Institution A, Roche analyzer) or as a qualitative assay (Institution B, Beckman Coulter analyzer), with 10 ng/mL as the manufacturer-specified cutoff for ARK POSITIVE, using manufacturer-supplied calibration standards ranging from 0-500 ng/mL X. Results The ARK semi-quantitative analysis (Roche analyzer) showed essentially equivalent curvilinear response curves for either X or 4-OHX at concentrations below 1000 ng/mL; for the semi-quantitative ARK assay, we use “&amp;gt;1200 ng/mL” for X-ARK &amp;gt;1200 ng/mL. By mixing studies (Roche), there were neither positive nor negative interferences observed as assessed for common drugs of abuse or for common therapeutic drugs. The presence of hemoglobin, bilirubin or moderate lipemia were devoid of assay interference. Precision studies for manufacturer-supplied controls were as follows: intra-assay CVs were 6.9% for the NEGATIVE control (5.1±0.35 ng/mL) and 5.9% for the POSITIVE control (15.7±0.53 ng/mL) (n=20); interassay CVs were 7.8% for the NEGATIVE control (5.1±0.40 ng/mL) and 5.7% for the POSITIVE control (15.9±0.90 ng/mL) (n=20). For the qualitative ARK assay as performed on the Beckman analyzer, POSITIVE and NEGATIVE controls ran invariably according to their designations. For the semi-quantitative assay application (Roche analyzer): among 78 X-NEGATIVE samples by LC-MS/MS, there were 0% ARK-POSITIVE results (false-positive rate=0%); among 74 X-POSITIVE samples by LC-MS/MS, there was one ARK-NEGATIVE result (false-negative rate=1.4%); this sample (X=65 ng/mL) was found to have a high amorphous crystalline content, and was unable to be reanalyzed. For the qualitative assay application (Beckman Coulter analyzer): among 78 X-NEGATIVE samples there was one ARK-POSITIVE sample (false-positive rate=1.3%) for a sample with xylazine=8.8 ng/mL by LC-MS/MS, with undetectable 4-OHX (&amp;lt;10 ng/mL); among 74 X-POSITIVE samples, there were no ARK-NEGATIVE results (false-negative rate=0%). Conclusions The ARK xylazine immunoassay demonstrated acceptable analytical performance with respect to detection of xylazine in urine at 10 ng/mL. With additional consideration of the assay’s cross reactivity with the major xylazine metabolite, 4-OH-xylazine, the assay was judged to be highly suitable for routine use as a screening test for detection of xylazine exposure via urine testing.

Comparison of estimated sodium intake by dietary recall method and spot urine samples in patients with CKD

Comparison of estimated sodium intake by dietary recall method and spot urine samples in patients with CKD

IL-6 and procalcitonin levels in hemodialysis patients with fungal infection

Patients with end-stage renal failure suffer from a defect in the immune system and a weakening of cellular and humoral immune defenses and thus an increase in the incidence of fungal infections. The aim of study is to isolate and to identify fungi from urine samples of patients with kidney failure. A study of relationship between fungal infection and the changes occurring in some immune parameters including Interleukin-6, Procalcitonin, high sensitivity-CRP in hemodialysis patients with and without fungi and the control group was made. This study was conducted on 190 samples of renal failure patients undergoing dialysis in the hemodialysis unit at Kirkuk Teaching Hospital during the period from November 2022 to the end of March 2023. Statistical analysis for immunological parameters was conducted using the Statistical Package for Social Sciences (SPSS). The results of laboratory culture showed the isolation of 58 fungal isolates, with a rate of 79.6% for yeasts and 20.4% for molds. Among the diagnosed yeasts, five types of yeasts belonging to the Candida genus showed different colors when cultivated on the differential medium Chrom Candida Agar (CCA) included Candida tropicalis, C. glabrata, C. parapsilosis, C. krusei and C. albicans. Immunological parameters of IL-6, PCT and hs-CRP showed higher levels in patients compared to the control group. The results also recorded an increase in the serum level of IL-6 in those with yeasts than in those with mold. Furthermore, there was also a level of PCT and hs-CRP in mortality compared to survivors patients. The study showed prevalence of fungal infections among patients of renal failure and identification species of fungi . Study of the signification differences in immunological parameters, such as IL-6 and procalcitonin, provide valuable insights into the associations between fungal infections and immune responses in patients.

Bioanalytical Method Development and Validation for the Estimation of Hydroxyproline in Urine Samples of Osteoarthritic Patients Using LC–MS/MS Technique

Bioanalytical Method Development and Validation for the Estimation of Hydroxyproline in Urine Samples of Osteoarthritic Patients Using LC–MS/MS Technique

TAM RECEPTORS IN THE PATHOLOGY OF BLADDER CANCER: APPROACHES TO PERSONALISED MEDICINE

Bladder cancer (BC) is a cancer of the lining epithelium of the bladder. In 75% of cases, it is presented as a non-muscle invasive disease (NMIBC), with a 5-year patient survival of more than 90%. However, 25% of BC patients develop an aggressive and chemoresistant muscle-invasive form of cancer (MIBC) with less than 50% 5-year survival, making BC the 13th deadliest cancer. Moreover, the treatment for MIBC, including cystectomy and bladder reconstruction, is costly and available only in developed countries, leaving patients in other countries without efficient treatment. So, there is an urgent need to develop and implement new personalised treatment schemes in Western and low – to middle-income countries with progressively ageing populations. In our work, we study the regulatory role of the TAM Receptor Tyrosine kinases (AXL, MERTK, and TYRO3) in signalling in BC. It has been shown before that TAM receptors are activated in different cancer types, where they regulate drug resistance, immune evasion and metastasis. We analyse how TAM signalling contributes to the development of chemoresistance in BC and propose that interfering with these pathways may be a part of new combination therapy approaches. We analyse TAM receptor expression and activation in primary patient materials (solid and liquid biopsies) and develop organoid cultures for testing anti-TAM inhibitors and chemotherapeutic agents. According to our recent data, soluble extracellular domains of TAMs are present in urine samples of BC patients and are suitable as urine predictive biomarkers to use in BC.

Urine culture Isolates and their sensitivity patterns.

Objective: To make an assessment of the susceptibility pattern to antimicrobials of various isolates, which would be helpful in deciding about further empirical treatment with antibiotics and would lead to improved patient outcomes. Study Design: Cross-sectional study. Setting: Watim Medical &amp; Dental College, Rawalpindi. Period: Jun 1, 2023 – Nov 30, 2023. Methods: 300 urine samples of patients suspected clinically to have urinary tract infection were directly collected directly from the microbiology laboratory, out of which 60 were culture positive. The positive samples comprised 44 females and 16 males. Results: The organism with the highest prevalence was Escherichia coli (E.coli), followed by Klebsiella species. Escherichia coli showed 100% sensitivity to tazobactam whereas Klebsiella species showed sensitivity to several different antibiotics. Conclusion: E.coli species was noted to be the most common organism causing UTIs. Antimicrobial resistance has been noted to be emerging against some antibiotics, and the current susceptibility patterns may be used in this area for achieving optimum therapeutic outcomes, and for preventing antibiotic misuse.