Sequential Application and Post-Test Probability for Screening of Bladder Cancer Using Urinary Proteomic Biomarkers: A Review based Probabilistic Analysis.

BACKGROUND: Bladder cancer is one of the most common cancers in the world, with men being affected more than women. Diagnosis by cystoscopy, cytology and biopsy is invasive. Urine cytology, a non-invasive modality is not sensitive. This study is undertaken to evaluate whether non- invasive urinary proteomic profiling is more sensitive, specific for bladder cancer. OBJECTIVE: To evaluate the sensitivity and specificity of various urinary proteomic biomarkers as a screening tool for bladder cancer. METHODS: PubMed database was searched from 4th December 2011 to 30th November 2021 using Mesh terms and n = 10,364 articles were found. PRISMA guidelines were followed and Review articles, animal studies, Urinary tract infections, non-bladder cancer and other irrelevant articles were excluded. All studies who have reported mean/median (SD/IQR), sensitivity, specificity, cut off values (ROC analysis) were included (n=5). Post-test probability of various biomarkers was calculated using sequential approach. Pooled analysis was depicted using Forest plot. RESULTS: Analysis of diagnostic studies of bladder cancer showed the post-test probability of CYFRA21-1 was 36.6%. Using sequential approach, the panel of biomarkers CYFRA 21-1, CA-9, APE-1, COL13A1 has post-test probability of 95.10% to diagnose bladder cancer. Analysis of two observational studies with APOE (n= 447) showed non-significant increase of APO-E levels in bladder cancer cases (WMD: 66.41with 95% CI 52.70-185.51; p=0.27, I2 92.4%). CONCLUSION: In patients presenting with hematuria, a panel of CYFRA 21-1, CA-9, APE-1, COL13A1 markers can be considered for screening of bladder cancer.

Risk factors for cardiovascular disease in a healthy young population: Family matters.

Background: India faces an epidemic of cardiovascular disease (CVD). This study sought the effect of family history of CVD and/or its risk factors (CVD-risk) on the presence of risk factors for CVD, in a healthy young college population. Methods: Blood pressure (BP), heart rate (HR), anthropometric variables, fasting blood sugar and lipid fractions were measured in two hundred healthy individuals (163 men and 37 women), aged 17-22 years. Data were analysed to elicit effect of CVD-risk on measured parameters. Results: All but one subject, had family history of a CVD-risk. Men with family history of coronary heart disease had higher diastolic BP (79.24 ± 7.7 vs 75.99 ± 7.49 mmHg, p = 0.007) and triglycerides (118.66 ± 57.98 vs 85.82 ± 50.89 mg/dL, p < 0.0001) compared with those without similar family history. Men with family history of hypertension (HTN) had higher diastolic BP (78.75 ± 7.15 vs 75.84 ± 8.37 mmHg, p = 0.019) and low-density lipoprotein (86.24 ± 25.38 vs 78.21 ± 17.93 mg/dL, p = 0.019), as well as lower high-density lipoprotein (50.27 ± 8.4 vs 53.96 ± 10.38 mg/dL, p = 0.019). Women with family history of diabetes mellitus had lower high-density lipoproteins (49.89 ± 8.05 vs 59.53 ± 11.44, p = 0.006). Family history of dyslipidaemia was associated with significantly higher triglycerides (146.14 ± 46.19 vs 98.44 ± 56.19 mg/dL, p = 0.002) in men and in subjects across sex. HDL was contrarily higher, in women with family history of cerebrovascular accident/HTN and men with family history of coronary heart disease/HTN. The proportion of pre-HTN, overweight/obese, impaired fasting glucose and borderline high triglycerides was 88.3%, 36.8%, 11% and 38.7% in men and 64.9%, 37.8%, 18.9% and 48.7% in female subjects. Conclusion: Young adults with a family history of CVD-risk already have an incomplete/atypical CVD risk profile.

A pilot study on DNA hypermethylation status in promoter region of P16 gene in patients with sporadic breast cancer.

Background: Epigenetic modification of cancer-related genes plays a role over and above their genetic alterations and contributes to the tumor initiation and progression of breast cancer. Promoter methylation of tumor suppressor genes is one such epigenetic modification, which can be potential biomarker. In this study, promoter methylation status of p16 gene was studied in blood samples of patients with breast carcinoma. Methods: Seventy-five patients, freshly diagnosed with carcinoma of breast and 20 age and sex matched healthy control subjects were recruited for the study. DNA extracted from EDTA blood sample was bisulfite converted and subjected to methylation-specific PCR to amplify the p16 promoter region. Results: Out of 75 patients, 25 (33%) patients showed hypermethylation in promoter region of p16 gene, which was statistically significant in comparison with the control group (p < 0.05). In subgroup analysis, lymph node involvement, cancer grade, and histopathological finding did not show any difference with methylation status of p16 promoter. Conclusion: Significant hypermethylation of p16 promoter region in the blood of histopathologically proven cases of breast cancer was observed suggesting promoter hypermethylation of p16 may be a possible mechanism accounting for sporadic carcinoma of breast.

Evaluation of a new interferon gamma release assay, in comparison to tuberculin skin tests and quantiferon tuberculosis goldplus for the detection of latent tuberculosis infection in children from a high tuberculosis burden setting.

Background: : Detection of latent tuberculosis infection (LTBI) in children, exposed to tuberculosis (TB) infections, followed by appropriate treatment, could be instrumental in reducing TB burden. Interferon Gamma release assays (IGRA) have higher specificity than tuberculin skin tests (TST), hence are more effective option for diagnosis. Hence, the present study was aimed to diagnose the presence of latent TB in children with the help of a new Standard E TB-Feron Enzyme-Linked Immunosorbent Assay (TBF) and evaluating its efficiency as compared to TST and Quantiferon TB Gold plus (QFT Plus). Methods: Diagnosis of LTBI in participants, <18 years old, who were the close household contacts of patients with active TB was carried out employing techniques such as TST, QFT Plus, and TBF. Results: Higher positive results were obtained with IGRA assays QFT Plus and TBF than TST. The perfect agreement was observed between QFT Plus and TBF assays with a Kappa value of 0.9176 whereas TST and TBF assay showed moderate agreement with a Kappa value of 0.42365. The level of cytokines secreted as a result of stimulation by the antigens from QFT Plus tubes was lower in comparison to antigens from TBF tubes. Conclusion: Hence, TBF, which showed similar efficiency as the widely used QFT Plus, can be a useful detection technique for LTBI in children. Moreover, it could prove to be an efficient alternative to expensive IGRAs like QFT Plus.

A study on DNA methylation status in promoter region of p15 gene in patients of acute myeloid leukemia and myelodysplastic syndrome.

BACKGROUND: Acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) are a spectrum of hematological malignancies with a multistep process of accumulated genetic and epigenetic alterations. DNA methylation is most extensively studied epigenetic alteration in malignancies. Recent research studies in the field have brought out translational implications of promoter methylation of tumor suppressor gene p15 in tumors. Therefore, we studied the role of DNA Methylation of p15 gene in AML and MDS. METHODS: The study was carried out in 41 consecutive AML/MDS cases reporting to hematological OPD of a tertiary care center along with 25 age and sex-matched healthy controls. The methylation status in the promoter region of the p15 gene was assessed by methylation-specific PCR (MSP) from blood samples after ethical approval and informed consent of the patients and controls. The association of methylation status was studied with clinical presentations, AML subtypes, and cytogenetics using Chi-square test/Fisher's exact test tools. RESULTS: A total of 41 cases included in the study comprised 33 cases of AML and 08 cases of MDS with an age range between 06 months and 82 years. Of the 41 cases, 29 revealed promoter methylation of the p15 gene, which compared to healthy controls was found statistically significant (p < 0.001). The methylation status did not significantly correlate with AML subtypes or the cytogenetic abnormalities detected in cases. CONCLUSION: The outcome of the study indicates p15 promoter DNA methylation in cases of AML and MDS may identify those individuals who might benefit from the targeted therapeutic approaches.

Sigma Metrics: A Valuable Tool for Evaluating the Performance of Internal Quality Control in Laboratory.

Background Six Sigma is a widely accepted quality management system that provides an objective assessment of analytical methods and instrumentation. Six Sigma scale typically runs from 0 to 6, with sigma value above 6 being considered adequate and 3 sigma being considered the minimal acceptable performance for a process. Methodology Sigma metrics of 10 biochemistry parameters, namely glucose, triglycerides, high-density lipoprotein (HDL), albumin, direct bilirubin, alanine transaminase, aspartate transaminase, urea nitrogen, creatinine and uric acid, and hematology parameters such as hemoglobin (Hb), total leucocyte count (TLC), packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and platelet were calculated by analyzing internal quality control (IQC) data of 3 months (June-August 2019). Results Sigma value was found to be > 6 for triglyceride, HDL, Hb, TLC, and MCH, signifying excellent results and no further modification with respect to IQC. Sigma value was between 3 and 6 for glucose, albumin, creatinine, uric acid, PCV, and MCHC, implying the requirement of improvement in quality control (QC) processes. Sigma value of < 3 was seen in AST, ALT, direct bilirubin, urea nitrogen, platelet, and MCV, signifying suboptimal performance. Discussion Six Sigma provides a more quantitative framework for evaluating process performance with evidence for process improvement and describes how many sigmas fit within the tolerance limits. Thus, for parameters with sigma value < 3, duplicate testing of the sample along with three QCs three times a day may be used along with stringent Westgard rules for rejecting a run. Conclusion Sigma metrics help assess analytical methodologies and augment laboratory performance.

To study the efficacy of thallium-201 as tumor seeking agent and to study its role in therapeutic response.

BACKGROUND: Tumour seeking characteristics of TL-201 have been underutilized. The study was undertaken to evaluate the role of TL-201 scintigraphy in tumour imaging. METHODS: A total of 50 cases were studied over a period of 18 months (13 lymphomas, 11 breast carcinoma, 10 lung cancer, 6 of soft tissue sarcoma, 2 bone tumours and 4 cases each of thyroid and brain tumours). Thallium-201 chloride was injected IV in the dose range of 3-5 mCi. Imaging was done using Siemen's ECAM dual headed gamma camera. Mean tumour to background ratio (T/B ratio) was calculated for all the positive cases. Descriptive statistical analysis was carried out. RESULTS: Findings revealed a sensitivity of 94.12%, specificity 87.5%, PPV 94.12%, NPV 87.50% and accuracy of 92% for TL-201 tumour imaging. The mean T/B ratio for the true positive cases at 10 min and at 3 h was 1.81 and 1.99 respectively, the difference being 0.18 (P value <0.001). Amongst them, the mean T/B ratios for low-grade tumours were 1.45 ± 0.32 at 10 min and 1.63 ± 0.38 after 3 h with difference of 0.176 (P < 0.001). For the high-grade tumours the ratios were 2.08 ± 0.35 and 2.26 ± 0.41 respectively with a difference of 0.186 (P < 0.001). CONCLUSION: Thallium-201 scintigraphy is a useful tumour imaging modality in cases of thyroid, breast, brain, lung, soft tissue and bone tumours and lymphomas. A T/B ratio of 1.63 ± 0.38 in 3 h-delayed imaging is suggestive of low-grade tumours. For high-grade tumours a ratio of 2.26 ± 0.41 should be considered significant.

Cost Effective Printing of Laboratory Reports: Biochemist's and Physician's debate and Institutions Dilemma.

INTRODUCTION: Laboratory investigations account for significant percentage of healthcare expenses. Every hospital and laboratory especially in the government sector wants to cut down the expenses on reports and offer inferior printouts of reports. The debate of printing attractive reports (clientele aspiration) Vs costs cutting (managerial issues) continues. It becomes the moral responsibility of doctors to guide the management into implementing an effective and economical option of report transcription. Cutting cost is always good but it should be achieved in an effective way keeping in mind the expectation of all the stake holders and ensuring that quality is not significantly compromised. The data regarding economics of printing available on the Internet may be market driven and not a true reflection of printing cost. The present study was undertaken to generate data on the economics of printing reports on various platforms to enable managers to make informed decision on choice of optimal printing option. AIM: To provide practical data on economics of printing reports using various printing options available in the country. MATERIALS AND METHODS: Four printing devices (laser printer, inkjet printer, dotmatrix printer and thermal printer) were connected to the laboratory information system and the reports printed from them were critically evaluated for the following parameters - cost of stationary, printing cost, time of printing and clientele satisfaction. RESULTS: The cost of printing a single report on laser printer, inkjet printer, dotmatrix printer and thermal printers was Rs 0.75, Rs 0.65, Rs 0.20 and Rs 0.93 respectively. The time taken to print 30 reports were 162 sec, 193 sec, 210 and 60 sec respectively. Clientele satisfaction scores were best with Laser and Inkjet printed reports. CONCLUSION: The most economical mode of printing is Dot Matrix printer using DOS output for printing. However, when the workload of a laboratory is high, then the use of laser printer is not prohibitively expensive option and can be resorted to. Use of thermal printer was found to be unsatisfactory.

Serum ferroxidase albumin ratio as a marken in pulmonary tuberculosis.

Serum ferroxidase and albumin levels were determined in 98 patients of tubercuiosis, of whom 49 were freshly diagnosed, sputum positive (group-I) & 49 were completely treated patients (group-II). Forty nine age and sex matched healthy individuals were taken as controls. Mean±SD of serum ferroxidase and albumin levels in controls, group-I and group-II was found to be 864.35±106.35 IU/L & 3.91±0.234 g/dL, 1603.76±222.65 IU/L & 3.24±0.518 g/dL and 1001.78±201.63 IU/L & 3.82±0.43 g/dL, respectively. Serum ferroxidase in group I was significantly higher as compared to controls and group-II (p<0.01). The decreased levels of serum albumin in group I, as compared to control and group-II was statistically significant (p<0.01). Serum ferroxidase: albumin ratio (Ferroxidase in International Unit per gram of albumin) in group I (50.47±10.36 IU/g) was significantly higher than controls (22.22±3.3 IU/g), (p<0.001) while in group II it was significantly lower (26.72±7.18 IU/g, p<0.001) than group-I and close to control values. Serum ferroxidase: albumin ratio (IU/g) can therefore be incorporated as a surrogate marker to assist in diagnosis and prognosis of pulmonary tuberculosis.