Objectives: Nasopharyngeal/oropharyngeal (NP/OP) swabs, the gold standard for COVID-19 diagnosis, pose challenges including patient discomfort, need for trained personnel, and biosafety risks. This study evaluated the diagnostic performance of saliva as a less invasive alternative compared to the gold standard. Materials and Methods: Paired NP/OP and saliva samples were collected from 515 suspected COVID-19 patients. NP/OP swabs underwent standard RNA extraction and reverse transcription-polymerase chain reaction (RTPCR). Saliva samples were analyzed by RT-PCR, either directly (saliva-direct [SD]) or after RNA extraction (saliva-RNA [SR]). A subset of discordant samples (saliva-positive, NP/OP-negative), with cycle threshold values ≤32, was investigated using Nanopore sequencing. Statistical analysis: Sensitivity, specificity, and Cohen’s Kappa were used to assess agreement between NP/OP and each saliva protocol. McNemar’s test compared detection rates between methods, and paired t-tests/Wilcoxon signed-rank tests compared Ct values between NP/OP and saliva-RNA assays. Results: Of 515 participants, NP/OP swabs yielded 114 positives (reference standard), SR yielded 117, and SD yielded 147. SR sensitivity/specificity was 61.4%/88.28% compared to NP/OP. SD sensitivity/specificity was 70.17%/83.29%. Cohen’s Kappa was 0.50 (0.399–0.581) for SR and 0.48 (0.395–0.569) for SD. 16.69% of saliva-positive samples were NP/OP negative. Nanopore sequencing confirmed severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) RNA in 78.26% of discordant saliva samples. Conclusions: Saliva shows promise as a complementary, adjunct specimen for COVID-19 diagnosis, potentially enhancing case detection. While sensitivity was lower than NP/OP, the direct saliva method demonstrated improved detection. The simplified collection and reduced biosafety risks associated with saliva offer significant advantages.
Objectives: This study compared the basophil counts (BC) on the automated analyzer Sysmex XN1000 (XN 1000) with manual differential count (DC). Materials and Methods: The study was conducted over a period of 2 years and included 77 diagnosed chronic myeloid leukemia (CML) cases. We compared the basophil percentages provided by XN 1000 with those of manual DC on peripheral blood smear (PBS). Manual DC was done by two hematopathologists on 200 cells, and the mean values were taken. Manual DC was considered the gold standard for comparison. Statistical analysis: Statistical analysis was done on R statistical software, version 4.1. Results: Seventy-seven patients of CML were included in the study. Mean basophil percentage (%) by XN1000 was 3.96% while the mean basophil % by manual DC was 7.07%. Pearson correlation between the two values was 0.249 with a significant p value (0.03). It was evident that the basophil percentage by automated analyzer was significantly lower compared to that obtained by manual counts. There was a mean difference (bias) of −3.11, suggesting that Sysmex XN 1000 can underestimate by ~3.1% when compared with the manual method. Conclusions: Automated analyzers (Sysmex XN 1000) underestimate the percentage of basophils in CML. The BC should be done manually on PBS for all cases of suspected CML since BC has a significant role in deciding the clinical management in CML patients.
Objectives: To evaluate analytical performance in a clinical biochemistry laboratory using an integrated approach combining variance index score (VIS), measurement uncertainty (MU), target score and root cause analysis (RCA) based on external quality assessment scheme (EQAS) data. Materials and Methods: This retrospective analytical study evaluated EQAS results generated over 2 years (January 2022 to December 2023) in a National Accreditation Board for Testing and Calibration Laboratories-accredited tertiary-care clinical biochemistry laboratory. A total of 1,853 EQAS results covering 68 analytical parameters were analysed across multiple platforms and programmes, including Bio-Rad EQAS, Randox International Quality Assessment Scheme and the Quality Assurance Forum. Analytical performance was assessed using VIS, target score for immunoassays, MU estimated by the Nordtest top-down approach and structured RCA for EQAS outliers. Statistical analysis: Descriptive statistics were applied. VIS, target score and MU were expressed as means with minimum and maximum values. EQAS outliers were reported as counts and percentages, and RCA findings were categorised by phase of error. Results: Of the 1,853 EQAS results analysed, 96.4% were within acceptable limits. Sixty-seven outliers (3.61%) were identified. Most deviations originated in the analytical phase (79.1%), followed by pre-analytical causes, while no post-analytical errors were observed. Routine chemistry, electrolyte and immunoassay parameters demonstrated predominantly good to excellent VIS and MU performance. Parathyroid hormone showed persistently high VIS and MU values, indicating analytical bias. Conclusions: The synthesis of VISs, MU, target scores and structured RCA offers a comprehensive framework for evaluating EQAS. Integrating these quality indicators streamlines the identification of analytical deviations, guides targeted corrective and preventive interventions and reinforces analytical reliability and continuous quality improvement within clinical biochemistry laboratories.
Objectives: The objective of the study is to evaluate the diagnostic performance of commercially available hepatitis A virus (HAV) immunoglobulin M (IgM) index tests, specifically enzyme-linked immunosorbent assays (ELISA) and rapid immunochromatographic tests (ICTs), against a chemiluminescence immunoassay (CLIA) serological reference standard. Materials and Methods: The diagnostic performance of four commercially available HAV IgM ELISA kits (Dia.Pro, MBS New S.R.L., Wantai, and Biogenix) and three rapid ICT kits (Biogenix, Biotrol, and Insight) was evaluated using a CLIA as the reference comparator. An initial assessment was conducted using 48 well-characterized external quality assurance serum samples, followed by evaluation of 250 residual clinical serum samples. Statistical analysis: Statistical analysis was performed using MedCalc Statistical Software. Sensitivity, specificity, positive predictive value, and negative predictive value were calculated with 95% confidence intervals. Inter-assay agreement with the reference comparator was assessed using Cohen’s kappa (κ) coefficient. A p <0.05 was considered statistically significant. Results: Among the ELISA kits, Dia.Pro and Wantai demonstrated optimal performance with 100% sensitivity and specificity and perfect agreement with the reference comparator. MBS New S.R.L. and Biogenix ELISA kits showed lower sensitivities of 92.0 and 84.8%, respectively, despite high specificity. Among rapid ICT kits, only the Insight assay showed complete concordance with the reference method, whereas Biogenix and Biotrol ICT kits demonstrated reduced sensitivity. Conclusions: Significant inter-assay variability exists among commercially available HAV IgM immunoassays. Independent evaluation and careful selection of diagnostic platforms are essential to ensure reliable laboratory diagnosis of acute HAV infection.
Objectives: Viral hepatitis (VH) due to hepatitis B and hepatitis C viruses (HBV/HCV) is a public health problem. Truenat provides molecular assays for VH detection. We aimed to inspect the successful and unsuccessful Truenat results and actions applied for reducing unsuccessful outcomes. Materials and Methods: Retrospective study (January 2024–December 2025) of secondary HBV and HCV Truenat data carried out at a district hospital in Una, Himachal Pradesh. Results reported as “detected,” “not detected,” “invalid,” and “error” (unsuccessful results). The laboratory adhered to standard operating procedures (SOPs) and good laboratory practices for decreasing unsuccessful results. Statistical analysis: Data extracted from the Truenat machine. Analyzed for the proportions of detected, not detected, invalid, and error. Difference between viral load values calculated with Student’s t -test. Results: A total of 537 plasma and 195 serum samples were evaluated for Truenat hepatitis B (n = 207) and hepatitis C viral load (n = 525). Average viral load for HBV and HCV was 5.9 × 10 7 IU/mL and 1.8 × 10 5 IU/mL, respectively. The mean viral load among plasma and serum was similar ( p = 0.213). In 2024 and 2025, the total rate of (HBV + HCV) detected was 57.7 and 62.1%, not detected was 42.3 and 37.9%. Meanwhile, the invalid and error rate was 17 and 12.7% in 2024, and in 2025, it was 2.5 and 5.1%, respectively. A reduction was noted in invalids and errors in 2025 compared to 2024. Conclusions: Truenat testing is advantageous in resource-poor settings. Plasma and serum samples compare favorably for viral load. Laboratories should follow good laboratory practices, identify reasons for unsuccessful outcomes, and resolve them through troubleshooting and precautionary measures.
Objectives: The primary objective of this study was to compare electrolytes and bicarbonate values obtained from arterial and venous blood samples in the same critically ill patients presenting to the emergency department (ED). The study also aimed at comparison of the turnaround time of point-of-care and laboratory auto-analyzer reports, aiding in early diagnosis and intervention. Materials and Methods: This was a cross-sectional observational study conducted in 100 critically ill patients presenting to the ED. From each patient meeting the inclusion criteria, both arterial and venous samples were collected simultaneously after ensuring quality control. Statistical analysis: Data analysis was done using Bland-Altman analysis, and plots were used to evaluate and visualize the degree of agreement between arterial and venous sodium, potassium, chloride, and bicarbonate. Results: The Bland–Altman plots demonstrated clinically acceptable agreement for sodium and bicarbonate, while chloride and potassium exhibited slightly higher variability. The negative bias (mean difference -0.36 mmol/L) in potassium indicates systematically higher venous levels. The intraclass correlation coefficient (ICC) analysis showed that ABG measurements are more consistent for sodium (ICC = 0.80, 95% confidence interval [CI] 0.70–0.87) and bicarbonate (ICC = 0.81, 95% CI 0.73–0.87), compared to chloride (ICC = 0.64, 95% CI 0.51–0.75). The mean turnaround time for ABG analysis was less compared to that of the central laboratory auto-analyzer (10 ± 2.5 min; 60 ± 12 min), making ABG a better point of care for electrolytes and bicarbonate analysis. Conclusions: This study showed that ABG, a point-of-care test with a shorter turnaround time when compared to an autoanalyzer, is helpful in providing rapid assessment of sodium and bicarbonate in the ED, while caution is needed for interpreting potassium and chloride.
Alveolar soft part sarcoma (ASPS) is a rare malignant neoplasm, accounting for <1% of soft tissue sarcomas. It typically presents as a slow-growing, painless mass, most often located in the deep soft tissues of the extremities. It is frequently associated with metastatic disease at the time of diagnosis. Distinguishing ASPS from its histological mimics can be challenging, particularly when it arises in atypical locations. We present a retrospective analysis of five cases of ASPS involving uncommon sites: breast, parapharyngeal region, mandible, elbow, and thigh. Histologically, all tumors exhibited a classic alveolar architecture, characterized by nests of polygonal cells separated by delicate vascular channels. Immunohistochemistry (IHC) confirmed transcription factor E3 nuclear positivity in each case, supporting the diagnosis. Awareness of such unusual presentations is essential to avoid misdiagnosis and inappropriate management. This series underscores the importance of careful histopathological evaluation and ancillary studies in recognizing ASPS at rare anatomical sites.
Congenital diaphragmatic hernia (CDH) is a relatively rare disorder in which the thoracic and abdominal compartments can communicate because the diaphragm does not fully form or fuse during embryologic development. Polycystic kidney disease (PKD) is a genetic disorder characterized by the formation of many renal cysts, which can affect kidney function. The co-occurrence of CDH and PKD in a fetus is highly unusual and provides major hurdles to prenatal diagnosis and management. This case report describes the prenatal diagnosis of CDH with PKD in the second trimester of pregnancy, highlighting the clinical presentation, imaging results, and fetal autopsy findings. The diagnosis was confirmed through detailed ultrasonography, which revealed herniation of abdominal organs into the thoracic cavity and the presence of cystic renal changes. Based on a literature review, there are very few documented examples of CDH and PKD coexisting. The degree of renal failure and the severity of pulmonary hypoplasia determine the prognosis in these cases. To maximize results, we discuss the significance of multidisciplinary management, including pediatric surgery, neonatology, and maternal-fetal medicine. This case contributes to the growing body of knowledge about rare fetal abnormalities and highlights the importance of comprehensive prenatal care and early diagnosis in informing clinical decision-making.
Extracellular vesicles (EVs) are increasingly recognized as active mediators of host–parasite communication rather than passive by-products of infection. Across protozoan and helminth parasites, EVs transport selectively packaged molecular cargo – including proteins, lipids, and nucleic acids – that modulate host immune responses, coordinate parasite populations, and contribute to disease pathogenesis. Emerging clinical and experimental studies highlight their translational relevance. In visceral leishmaniasis, proteomic analysis of plasma-derived EVs has identified host proteins that distinguish active disease from post-treatment states, while in malaria, EV-associated RNA cargo released from infected erythrocytes reflects parasite developmental dynamics and disease severity. Helminth-derived vesicles similarly carry conserved microRNAs and immunomodulatory molecules capable of altering host inflammatory pathways. These findings position parasite EVs as potential biomarkers for infection staging, treatment monitoring, and relapse surveillance. Experimental investigations further suggest that vesicle biology may be harnessed for therapeutic purposes. EV-associated antigens have shown partial protective efficacy in vaccination models, and engineered or hybrid vesicle systems are being explored as delivery vehicles for therapeutic cargo. At the same time, pharmacological disruption of vesicle biogenesis or uptake has emerged as a potential strategy to interfere with parasite communication pathways. Despite these advances, translation into clinical practice remains limited. Most studies rely on small discovery-stage cohorts and heterogeneous analytical methods, highlighting the need for standardized isolation protocols, validated biomarkers, and multicenter clinical evaluation. Integration of emerging technologies – including single-vesicle profiling, advanced imaging, and multi-omic approaches – may further clarify the biological functions and diagnostic value of parasite-derived EVs. Together, current evidence indicates that EVs represent a promising interface between parasite biology and translational innovation. Continued progress in methodological standardization, biomarker validation, and scalable therapeutic strategies will be essential to realize the potential of EV-based diagnostics and interventions for parasitic diseases.
Objectives: The objective of the study is to evaluate the concordance, turnaround time (TAT), and clinical implications of direct antimicrobial susceptibility testing (AST) performed from positive BacT/ALERT ® blood culture bottles compared with conventional AST. Materials and Methods: A prospective study was conducted in the Microbiology Department of UCMS and GTB Hospital, including 250 positive blood cultures from patients with suspected bloodstream infections. Direct AST and conventional AST were performed according to Clinical & Laboratory Standards Institute (CLSI) guidelines. Categorical agreement (CA), TAT, and error rates were assessed. Statistical analysis: Descriptive statistics were used. Continuous variables (TAT) were compared using the Student’s t -test. CA between methods was evaluated using the Chi-square test with degrees of freedom and p values reported. Results: Enterobacterales showed 94.4-99.4% CA, and non-fermenters demonstrated 92.8-100% agreement, with all error rates within CLSI-acceptable limits. Direct AST reduced mean TAT from 28 h (range: 18-32 h) to 9.7 h (range: 8-12.5 h). Direct AST demonstrated the greatest reduction in turnaround time in bloodstream and respiratory infection samples. Conclusions: Direct AST demonstrated high concordance and significantly reduced TATs compared with conventional AST. It enables earlier targeted therapy and supports antimicrobial stewardship.
Objectives: Urinalysis remains a fundamental diagnostic tool in the evaluation of renal and urological disorders. Differentiation between glomerular and non-glomerular hematuria based on red blood cell (RBC) morphology is critical for clinical decision-making. While phase-contrast microscopy (PCM) is considered the reference standard for identifying dysmorphic RBCs (dRBCs), its limited availability and operator dependency necessitate assessment of alternative methods. This study aimed to compare light microscopy (LM), PCM, and the DIRUI FUS-1000 automated urine analyzer for detecting dRBCs and identifying glomerular hematuria. Materials and Methods: This cross-sectional study included 150 patients presenting with hematuria. Fresh urine samples from each patient were examined independently using LM, PCM, and the DIRUI FUS-1000 automated urine analyzer. The origin of hematuria (glomerular or non-glomerular) was determined using a composite reference standard incorporating clinical findings, laboratory investigations, imaging studies, and histopathological data where available. Statistical analysis: Diagnostic performance parameters, including sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy, were calculated for each modality. Receiver operating characteristic (ROC) curve analysis was performed to assess diagnostic discrimination and determine optimal cut-off values for dRBC percentages. Statistical analysis was conducted using the Statistical Package for the Social Sciences version 22.0. Results: Of the 150 cases analyzed, 102 (68%) were classified as having glomerular hematuria and 48 (32%) as non-glomerular hematuria. PCM demonstrated the highest diagnostic accuracy, with a sensitivity of 100%, specificity of 75%, and an area under the ROC curve (AUC) of 0.97 at a cut-off value of 21% dRBCs. LM showed moderate diagnostic performance (sensitivity 77.4%, specificity 56.3%, and AUC 0.76). The automated urine analyzer demonstrated comparable sensitivity to LM but lower specificity, with an AUC of 0.72. Conclusions: PCM remains the reference standard for identifying dRBCs and diagnosing glomerular hematuria. However, LM and automated urine analyzers serve as practical screening tools, particularly in high-volume laboratory settings. Emerging artificial intelligence-assisted digital urine microscopy platforms hold promise for improving reproducibility, diagnostic accuracy, and workflow efficiency in routine urinalysis.
A 47-year-old woman presented with acute intestinal obstruction. Contrast-enhanced computed tomography suggested an ileocecal stricture, and an emergency right hemicolectomy was performed. Grossly, an annular stricture with intact mucosa was identified. Histology showed diffuse mural infiltration by discohesive tumor cells arranged in cords and single-file strands with mucosal sparing, favoring metastatic invasive lobular carcinoma (ILC). Targeted review revealed a neglected right-breast mass, core biopsy showed hormone-receptor-positive, human epidermal growth factor 2-negative carcinoma with lobular morphology, and low Ki-67. Immunohistochemistry demonstrated breast lineage (GATA3-positive) and aberrant/retained E-cadherin with p120 catenin cytoplasmic redistribution (with residual membranous staining) and loss of membranous β-catenin, supporting ILC with aberrant E-cadherin expression rather than ductal carcinoma. The patient was referred for systemic therapy. This case illustrates a “double deception,” a clinical mimic of ileocaecal tuberculosis or primary colonic carcinoma and a pathological pitfall due to deceptively retained E-cadherin staining, underscoring the risk of misdiagnosis and the critical importance of careful microscopic morphology and a high index of suspicion, supported by a morphology-first, panel-based immunohistochemical approach (E-cadherin/p120/β-catenin) when E-cadherin expression is aberrant.
Objectives: Microsatellite instability (MSI) is commonly associated with various malignancies, including colorectal, gastric, and endometrial carcinomas. The study aims to investigate the prevalence of mismatch repair (MMR) protein deficiency in the subtypes of renal cell carcinoma (RCC). Materials and Methods: Immunohistochemistry was done on 50 cases of RCC for four MMR proteins (MLH1, MSH2, MSH6, PMS2). The results were interpreted based on the percentage and intensity of expression, and the cases were categorized as MMR-proficient and MMR-deficient (dMMR). These were then correlated with clinicopathological parameters. Statistical analysis: One-way analysis of variance followed by least significant difference post hoc analysis; the Chi-square test was done to find the association between variables. The probability value ( p < 0.05) was considered statistically significant. Results: There were 50 cases of RCC. The male-to-female ratio was 3.5:1. Clear cell RCC was the most common subtype. 28% ( n = 14) of the study cases were dMMR. The most frequent expression pattern among dMMR cases was combined loss of MSH2 and MSH6 (12%), followed by a complete loss of all 4 markers (10%). There was a significant correlation of the expression pattern of MMR proteins with the tumor grade, renal vein/sinus involvement, and perinephric fat invasion. Conclusions: MMR deficiency was observed in 28% of RCC cases. The association between MMR status and tumor grade, renal vein/sinus involvement, highlights the clinical relevance of MMR analysis in RCC. These findings contribute to our understanding of the molecular underpinnings of RCC and may inform future diagnostic and therapeutic strategies.
Objectives: The objectives of the study are to primarily assess changes in LCS during the intensive phase of anti-TB therapy and secondarily to examine associations between LCS and microbiological and inflammatory indicators and high-sensitivity C-reactive protein [hsCRP] and to explore whether baseline LCS predicts sputum culture conversion at 2 months. Materials and Methods: This was a prospective observational cohort study conducted at a single tertiary-care. Fifty-six treatment-naïve adults (≥18 years) with microbiologically confirmed pulmonary TB were enrolled at treatment initiation. LCS (range 3-21) was assessed at baseline and after the 2-month intensive phase. Sputum smear microscopy and Löwenstein-Jensen culture were used to assess bacteriological status, and baseline hsCRP was measured. Statistical Analysis: Continuous variables were summarized as mean ± standard deviation or median (interquartile range [IQR]). Paired comparisons used the paired t-test or Wilcoxon signed-rank test, and between-group comparisons used non-parametric tests as appropriate. Associations were assessed using Spearman’s correlation. Receiver operating characteristic (ROC) analysis was performed to evaluate baseline LCS for predicting 2-month sputum culture conversion. Results: Median LCS improved from 9.75 (IQR 7.60–10.90) at baseline to 17.0 (IQR 15.72–17.77) at 2 months (Z = -6.151, p <0.001). Median hsCRP declined from 42.56 mg/L to 8.64 mg/L ( p <0.001). Sputum culture conversion occurred in 47 of 56 patients (83.9%). Baseline LCS did not differ significantly between converters and non-converters ( p = 0.101) and correlated poorly with baseline hsCRP (r = −0.14, p = 0.303). In contrast, follow-up LCS and improvement in LCS were significantly greater among patients who achieved sputum conversion. Conclusions: LCS is a highly responsive and feasible patient-reported outcome measure that captures clinically meaningful symptomatic improvement during the intensive phase of anti-TB therapy. However, baseline LCS shows limited association with microbiological burden or systemic inflammation and should not be used as a stand-alone predictor of early sputum culture conversion.
Objectives: Urinary protein or albumin can be an early laboratory modality to diagnose nephropathy early in rheumatoid arthritis (RA). Therefore, our objective was to estimate the prevalence of microalbuminuria in RA and to find any association with disease activity and other parameters. Materials and Methods: We conducted this cross-sectional study from 2022–2023. It included 75 RA patients aged ≥18 years. Urine albumin-to-creatinine ratio (ACR) was measured in all participants. All clinical and laboratory parameters were recorded. Disease activity was measured by the disease activity score-28 erythrocyte sedimentation rate (ESR). Statistical analysis: Appropriate statistical tests were used by the Statistical Package for the Social Sciences version 22. Pearson’s correlation was used to determine the correlation between microalbuminuria and disease activity and other factors. Results: The mean age of participants was 45.70 ± 11.43 years. The majority of participants had moderate to high disease activity (81.3%). Microalbuminuria was seen in 13.3%, whereas 9.3% participants had macroalbuminuria. Microalbuminuria correlated significantly with disease activity ( p = 0.032) and C-reactive protein (CRP) ( p = 0.004) but not with ESR ( p = 0.197). Furthermore, albuminuria correlated significantly with the duration of disease (r = 0.87, p = 0.011). Microalbuminuria was less common in those who were on combination disease-modifying anti-rheumatoid drugs (DMARDs) ( p = 0.045). Conclusions: The prevalence of microalbuminuria and macroalbuminuria was high in the Indian cohort. Furthermore, a higher degree of microalbuminuria correlated with disease severity and longer duration of disease. However, a lower incidence of albuminuria was associated with the use of a combination of conventional DMARDs. Therefore, emphasis should be placed on periodically monitoring the urine ACR, especially in advanced active disease.
Objectives: The objective of the study is to evaluate regulatory T-cell (Treg) levels in pediatric B-cell acute lymphoblastic leukemia (B-ALL) at diagnosis and after induction chemotherapy and to assess their association with clinico-hematological parameters and treatment response. Materials and Methods: This prospective observational study was conducted from October 2023 to March 2025 and included 39 children (<18 years) newly diagnosed with precursor B-ALL. Peripheral blood Tregs were quantified at baseline and after induction chemotherapy using flow cytometry with CD45, CD4, CD25, and Foxp3 markers. Clinical characteristics, hematological parameters, risk stratification, and treatment response were documented. Statistical analysis: Comparisons between baseline and post-induction values were performed using the Wilcoxon signed-rank test. Associations between variables were analyzed using Spearman correlation, Kruskal–Wallis test, and Mann–Whitney U test. A p <0.05 was considered statistically significant. Results: Treg levels showed a significant reduction following induction chemotherapy ( p <0.001). Post-induction Treg levels were significantly higher in females compared with males ( p = 0.044). CD4 + T-cell levels increased significantly after induction therapy ( p = 0.0313). No significant associations were observed between Treg levels (baseline, post-induction, or change) and age, hematological parameters, risk category, or treatment response. Conclusions: Induction chemotherapy results in a significant decline in Treg levels in pediatric B-ALL, with a notable gender-based difference observed after treatment. Larger longitudinal studies are warranted to determine the potential role of Tregs as biomarkers and therapeutic targets in B-ALL.
Objectives: The main objective of the study is to correlate immunohistochemistry (IHC) expression of Her2neu, p53, and Ki67 with clinicopathological parameters in invasive urothelial carcinoma (UC) cases. Materials and Methods: Sixty cases of invasive UC were included in the study. IHC for Her2neu, p53, and Ki67 was performed in all these cases and correlated with clinicopathological parameters such as age, sex, smoking history, grade, and stage of tumor. Statistical analysis: Performed using SPSS software version 11.5 (SPSS, Chicago, IL, USA), and the Chi-square test was used to compare the variables. A p <0.05 was considered statistically significant. Results: The mean age was 61 years, with the majority of patients being male (96.6%). Half of the patients were smokers, and the majority of cases were high-grade UC (73.3%). A statistically significant correlation was noted between Her2neu, p53, and Ki67 expression with grade and stage of tumor ( p <0.05). There was no significant correlation between these IHC expressions and age, sex, and smoking history of patients. Furthermore, it was noted that a statistically significant correlation was also noted between positive IHC expression of Ki67 with Her2neu and p53. Conclusions: The expression of Her2neu, p53, and Ki67 is more prevalent in invasive UCs of higher grade and stage. Their expression in these cases can suggest a possible role in the clinical prognostication of these cases.
Pharmacokinetic-pharmacodynamic (PK-PD) targets underpin antifungal dosing and therapeutic drug monitoring (TDM), but most were derived from animal models, small clinical cohorts, and wild-type susceptibility distributions that no longer reflect contemporary resistance. Static thresholds such as voriconazole trough ranges or echinocandin area under the curve/minimum inhibitory concentration (MIC) ratios improved outcomes in susceptible populations but are increasingly brittle against resistant phenotypes. Rising azole resistance in Aspergillus fumigatus , driven both by long-term therapy and agricultural fungicide exposure, and the global epidemiological shift in candidemia toward Candida glabrata and Candidozyma auris have reshaped the MIC landscape. Clinical paradoxes now emerge: Breakthrough infections despite “therapeutic” drug levels, toxicity when escalating doses to chase higher MICs, and poor outcome prediction once FKS mutations undermine echinocandin activity. This review synthesizes historical foundations of antifungal PK-PD targets, highlights how evolving MIC distributions deform the probability of target attainment, and analyzes why genotype rather than MIC alone predicts clinical failure in C. glabrata . It critiques the limitations of current breakpoints, explores special populations in which PK variability magnifies risk, and emphasizes how diagnostic and methodological gaps perpetuate miscalibration. Moving forward, recalibration should link surveillance to modeling through minimal interoperable datasets, embed real-time PK-PD dashboards in TDM laboratories, and incorporate genotype-informed decision rules. Region-specific MIC distributions, site- and biofilm-aware targets, combination PK-PD indices, and next-generation antifungals such as rezafungin and ibrexafungerp must be integrated into new frameworks.
