Multicentric evaluation of conventional dosimetry vs bio-dosimetry over a period of two years for a three-point contact.

Background: Dental radiology represents the best model for evaluating the effects of low-dose ionizing radiation. Therefore, this study evaluated the awareness on radiation hygiene among dental ancillary personnel through a questionnaire and their absorbed doses by physical and biologic dosimetry. Methods: The multicentric study included two groups. Group I (N = 30) consisted of dental staff involved in dental radiology. An equal number of personnel who were not related to radiology formed the control group. Knowledge (K), attitude (A), and practice (P) of participants were assessed using a KAP questionnaire. Radiation exposure was evaluated by physical dosimetry at 3 time periods: at the beginning of the study (T1), after 10 months (T2), and at the end after 20 months (T3). Similarly, biologic dosimetry was also carried out at 3 time points by dicentric chromosome aberration assay. The data were compared using percentage analysis, analysis of variance (one-way analysis of variance), and Student's t- test. Results: The KAP survey demonstrated enhanced understanding of radiation protection measures and its sound practice by the participants. Physical dosimetry showed a significant increase in absorbed dose at 3 time points: T1, T2, and T3. However, no chromosomal aberrations were observed in blood lymphocytes for any of the participants in the optimized 4-day biodosimetry protocol. Conclusion: Good radiation protection protocols-safe distance from the radiation source and wear of lead aprons and thyroid collars-ensured low absorbed doses. The 4-day protocol is an important step toward developing biodosimetry laboratories in the Armed Forces Medical Services for clinical and national radiation countermeasure strategies.

A comparison of the efficacy of narrow band imaging and contact endoscopy in an early diagnosis of squamous malignancies of the upper aerodigestive tract.

Background: This study aims to compare the efficacy of narrow band imaging (NBI) endoscopy and contact endoscopy in early diagnosis of squamous malignancies of upper aerodigestive tract. Methods: This study was of 18 months duration, sample size 50, and carried out at tertiary care hospital. The patients were subjected initially to NBI endoscopy followed by contact endoscopy. Thereafter, the lesion was biopsied and subjected to histopathological examination as is done routinely. The images obtained were analyzed based on criteria proposed by earlier studies and compared with histopathological examination as gold standard. Results: The sensitivity, specificity, and negative predictive values of NBI in early diagnosis of squamous malignancies of upper aerodigestive tract were high and better than contact endoscopy. Conclusion: Endoscopic NBI is a noninvasive and promising tool used for in vivo differentiation between malignant and nonmalignant lesions of upper aerodigestive tract by using morphology of mucosal capillaries and is more efficacious than contact endoscopy. It can be employed as part of routine ENT examination in outpatient departments; however, it has got a learning curve associated with it.

Rapid detection of carbapenem resistance among gram-negative organisms directly from positive blood culture bottles.

Background: Carbapenemase producing gram-negative bacteria (GNB) has become a huge problem in majority of tertiary care centers worldwide. They are associated with very high morbidity and mortality rates, especially when they cause invasive infections. Therefore, rapid detection of these organisms is very important for prompt and adequate antibiotic therapy as well as infection control. The aim of this study was rapid detection of carbapenemase genes and thereby likely carbapenem resistance, 24-48 hours in advance, directly from the positive-flagged blood culture bottles using CHROMagar and Xpert® Carba-R. Methods: Aspirate from positively flagged blood culture bottles was subjected to differential centrifuge. All gram-negative bacilli on gram stain from the deposit were processed in Xpert® Carba-R and inoculated on CHROMagar. The presence of genes and growth on CHROMagar was compared with carbapenem resistance on VITEK-2 Compact. Results: A total of 119 GNB isolates were processed. One or more of the carbapenemase genes were detected in 80 isolates. On comparison with VITEK-2 result, 92 samples showed concordance for carbapenem resistance 48 hours in advance. There was discordance in 21 isolates with 12 major errors and 09 minor errors. The sensitivity of direct Xpert® Carba-R test for rapid detection of carbapenem resistance, 48 hours in advance, was 81.42%. The sensitivity of direct CHROMagar test for accurate detection of carbapenem resistance, 24 hours in advance, was 92.06%. Conclusion: The ability to detect carbapenem resistance with very high accuracy, 48 hours in advance, helps in appropriate antibiotic therapy and implementation of effective infection control practices.

Cancer: A pathologist's journey from morphology to molecular.

The journey of cancer has taken significant positive steps towards better diagnosis and management. Pathologists have taken a leading role in this journey. Started with morphology which itself went through substantial developments in knowing the aetiology, pathogenesis, classification based upon the pattern, cellular details of the cancer. However, the histomorphologists met some road blocks in the form of ambiguities and some limitations that led to the expansion of pathologist's field to various other ancillary tests to enhance its values form diagnosis to therapeutics. This new journey facilitated the confidence of pathologist in the diagnosis, expanded their role in the clinical management through targeted therapy, monitoring of drugs, suggestions of prognosis. Various molecular diagnostic tests have been developed for the identification of tumor molecular profile for exploration of targeted therapy. The major problems in the management of cancer were resistance to chemotherapy, relapses and wide-spread side effects of chemotherapy. All these major problems to some extents are getting resolved with time as the molecular profiling is opening the gates. Many questions are being answered with the evolution in pathology. With the revolution of targeted therapy many difficult cancers are being treated with fewer side effects. In this review we will go through the journey from histomorphology, its advancement, digital pathology to the molecular pathology. These advancements interestingly haven't replaced the other, rather have enhanced others for the better management of cancer.

Evaluating morphological features for predicting microsatellite instability status in colorectal cancer.

Background: Colorectal cancer (CRC) is one of the commonest cancers worldwide, with incidence rates in India being around 4%. It is a heterogeneous disease with multiple established prognostic factors. Ten to fifteen percent originate from microsatellite instability (MSI) pathway, characterized by defect in mismatch repair (MMR) gene. Identification of MMR defective protein is relevant for diagnosis, prognosis, and prediction. Certain clinical and histological features are known to be associated with defective MMR genes. The objectives of this study are to find the prevalence of MSI in CRC to identify features associated with MSI and assess the value of histopathology in predicting MSI. Methods: We evaluated various clinical and histological parameters for identifying prognostically favorable colon cancers in a tertiary hospital. One hundred fifty colon cancers were evaluated, and MSI status was correlated with clinicopathologic variables. Results: The prevalence of MSI in CRC was found to be 11.3%. The factors associated with MSI were tumor differentiation, stage, tumor site, tumor size, tumor-infiltrating lymphocytes, Crohn's-like lymphoid reaction, and dirty necrosis. We have defined a "P" score for prediction of MSI using the clinicohistological parameters, which could be used to select patients who are to be tested for MSI. Conclusion: Assessment of clinical and histopathological features will help in patient stratification and selection of patients for MSI testing. The evaluation is economical, reproducible, and easy to apply.

Detection of dengue virus serotypes by single-tube multiplex RT-PCR and multiplex real-time PCR assay.

Background: All four dengue serotypes cause infection, with one of them predominantly reported from a particular geographical region. Coinfection by more than one serotype is reported from hyperendemic regions. These coinfections are clinically more severe than infection with a single serotype. This study was carried out to detect the predominant dengue serotype and presence of coinfections. Methods: Acute-phase serum samples of patients suffering from dengue infection were collected. They were screened for the presence of IgM, IgG and NS1Ag by a rapid test. Conventional multiplex reverse transcriptase polymerase chain reaction (RT-PCR) and multiplex real-time RT-PCR assays were carried out for detection and serotyping of the dengue virus. Results: A total of 196 samples were positive by the rapid card test. Of these, 139 were NS1Ag positive, 40 were positive only for IgM, 5 were positive only for IgG and 12 samples were positive for different combinations of antigen and antibodies. All four serotypes were detected in these samples by PCR. DENV-3 was found to be most common circulating serotype. A total of 22 cases were found to have coinfection with more than one dengue serotypes. Samples having only antibodies and no antigen on rapid card test were also positive for virus by PCR. Conclusion: Prevalence of dengue co-infections is increasing. Moreover, it is important to screen for dengue virus in those samples also which do not show NS1Ag on rapid tests and have either one or both the antibodies. Real-time multiplex RT-PCR is found to be more sensitive in detecting coinfection than conventional multiplex RT-PCR.

Characteristics, therapeutic modalities and outcomes of COVID-19 ventilated patients in a tertiary care hospital in counter-insurgency zone: Our experience.

BACKGROUND: Coronavirus disease 2019 (COVID-19) can result in severe life-threatening course requiring ventilatory support. This study highlights data pertaining to ventilated patients to enhance our understanding of COVID-19 as it evolves. METHODS: A descriptive, retrospective analysis was carried out on 50 COVID-19 RT-PCR positive patients who received mechanical ventilation at a tertiary care hospital in counter-insurgency (CI) zone, from June to December 2020. Data pertaining to patient characteristics, treatment, ventilator support and outcomes was analysed. RESULTS: Out of 50 patients, 74% were aged 50 years and above with 60% patients having comorbidities. 39 patients received non-invasive ventilation (NIV) and 04 patients received invasive mechanical ventilation (IMV) while 07 patients were converted from NIV to IMV during the hospital stay. Out of the 50 patients who received ventilator support 25 (50%) survived to discharge. The overall survival was 47.3% amongst the males while it was 58.3% for the females. The majority of survivors were in the NIV category (61.5%) while only 9.0% survived amongst those who received IMV. Average length of stay on NIV for patients was 5.3 days and for IMV was 7.5 days. All 50 patients received therapy in the form of steroids, anticoagulants, broad spectrum antibiotics and antivirals. Remdesivir was given to 40 of these patients out of which 20 survived (50%). Interleukin-6 therapy (Tocilizumab) was given to five patients of which four survived (80%). CONCLUSION: This study helps us to gain insights into the outcomes of COVID-19 patients managed in a tertiary care hospital in CI zone.

Keeping in pace with the new Biomedical Waste Management Rules: What we need to know!

Biomedical Waste Management Rules were first implemented in our country on 20th July 1998. Thereafter, the rules have undergone periodic updates and amendments in the years 2003 and 2011. Latest Biomedical Waste Management Rules, 2016, and (Amendment) Rules, 2018, were an update and simplification of BMW disposal as compared with the previous version, keeping in pace with the changes in the requirements of the health-care setup. Although exhaustive, numerous medical devices/products/kits did not find any mention even in the latest amendment of the rules. Thus, this article aims to bring out the key points to be known by all health-care workers and the gray areas which require clarification and inclusion in the rules for a completeness of the said rules.

Direct antimicrobial susceptibility testing from positive blood culture bottles in laboratories lacking automated antimicrobial susceptibility testing systems.

BACKGROUND: Timely initiation of appropriate antimicrobial can improve the outcome in terms of reduced morbidity and mortality in addition to reduced health-care costs. Availability of early preliminary Antimicrobial Susceptibility Test (AST) report will be useful in directing antimicrobial therapy. The aim of the study was to correlate AST by disc diffusion method, directly from positively flagged blood culture bottles, with the AST by automated method. METHODS: A total of 144 aerobic blood culture bottles flagged positive by the automated blood culture system were processed. The bacteria were pelleted by two-step centrifugation of the broth from the bottle and used to make a smear for Gram stain as well as an inoculum for antimicrobial sensitivity testing by Kirby Bauer disc diffusion method. Automated identification and AST were also carried out. RESULTS: On direct staining, 94 samples showed gram-negative bacilli, 39 showed gram-positive cocci, and 11 showed yeasts or polymicrobial growth. In the case of gram-negative bacteria, there was 99% categorical agreement between direct sensitivity testing and automated sensitivity testing with 1% disagreement. Among the gram-positive cocci, there was 96% categorical agreement with 4% disagreement between the two methods. CONCLUSION: High degree of agreement between the two methods is promising and applicable to situations where automated sensitivity testing is not available. Even if the systems are available, this method would prove useful as an adjunct to standard AST reporting. This sensitivity report can be generated earlier than the conventional AST, enabling choice of appropriate antimicrobial.

Molecular characterization and phylogenetic analysis of Chikungunya virus from Delhi, India.

BACKGROUND: Chikungunya virus is an alpha virus with high similarity to Dengue and Zika viruses, both in transmission cycle and in clinical presentation. Chikungunya is a re-emerging mosquito-borne infection known to cause small to very large outbreaks/epidemics at frequent intervals. In 2016, India witnessed a large outbreak of Chikungunya infection affecting more than 58,000 people. This study was undertaken to look at the genotypic phylogeny to know the relatedness with previously reported strains. METHODS: During the 2016 outbreak, samples from all patients clinically suspected to have Chikungunya were collected and subjected to testing for IgM antibody by ELISA and viral RNA detection by RT-PCR. Sequencing of the E1 gene segment was done to create a phylogenetic tree comparison with other strains. RESULTS: Serum samples were collected from 142 patients of clinically suspected Chikungunya infection. Majority of the patients were in the age group of 31-50 years accounting for more than 35% of the total cases. Twenty eight samples were positive for IgM antibody. Thirty seven samples were positive for viral RNA by RT-PCR. Only 06 cases were positive by both tests. Mutations in the amino acids K211E, M269V and D284E in the E1 gene segment of the Chikungunya virus were observed in the seven strains that were sequenced. On phylogeny tree, all the strains were found to belong to the ECSA genotype. CONCLUSION: Actively searching for the potential epidemic causing mutations and reporting of novel mutations may help in better understanding and probably forecasting of future CHIKV outbreaks and its nature.

Pre-eclampsia: Molecular events to biomarkers.

Pre-eclampsia is a hypertensive disorder in pregnancy, which accounts for 10-15% of the maternal and perinatal mortality worldwide. Abnormal placental development and tissue hypoxia are its main etiologic factors. The present diagnostic methods of blood pressure monitoring and renal function evaluation are insufficient in the early detection of pre-eclampsia. Since molecular events portent well ahead of the disease onset, biomarker research for the early diagnosis of pre-eclampsia has recently generated ambitious clinical targets. However, no clinically validated biomarker has so far been reported for the prediction of pre-eclampsia. Therefore, this review takes stock of the current understanding of pre-eclampsia from a molecular biology perspective and critically evaluates the following diagnostic potentials claimed for the biomarkers: placental proteins, angiogenic markers, and cell-free fetal DNA (cffDNA) in maternal circulation. Though the emerging evidences in favor of the fetal-specific epigenetic marker, hypermethylated RASSF1A of cffDNA, are highlighted, it pitches for a broader strategy of 'combination biomarker approach' for the reliable forecasting and triaging of pre-eclampsia.

Report on the newly emerging nosocomial Burkholderia cepacia in a tertiary hospital.

BACKGROUND: Burkholderia cepacia is an aerobic, motile, opportunistic Gram negative bacillus that can survive in certain disinfectants. This is a report of the emerging infection with the bacteria B. cepacia in our hospital. The awareness of this emerging bacterium is important, as it is known to cause nosocomial infection in hospitals, especially in the Intensive Care Unit (ICU) setting. setting. B. cepacia, although known to be multidrug resistant, shows sensitivity to some antibiotics that can be used to treat infection caused by it. METHODS: The cases of infection and antimicrobial susceptibility of nosocomial B. cepacia pattern have been analyzed. RESULTS: A total of 38 cases with B. cepacia infection were isolated. Two of these cases showed the organism in two samples, totalling the sample collection to 40. The most frequent isolation of B. cepacia was from blood 21/40 (52.5%) and pus 9/40 (22.5%). B. cepacia infections were most commonly observed in the Intensive Care Unit (52.6%). Infections were more common in men than women with a mortality rate of 42%. The most sensitive antimicrobial agents were found to be Colistin (93%) and Cotrimoxazole (71%). CONCLUSION: There have been 38 cases of the emerging nosocomial B. cepacia infection in our hospital in the period from September 2012 to February 2014. There was no case reported in the records before September 2012. Infections caused by B. cepacia should be made aware of and taken seriously because of its high transmissibility, intrinsic resistance to antibiotics, high mortality and most importantly its sensitivity to simple antibiotics such as Cotrimoxazole.

Laser ionization mass spectrometry in oral squamous cell carcinoma.

Biomarker research in oral squamous cell carcinoma (OSCC) aims for screening/early diagnosis and in predicting its recurrence, metastasis and overall prognosis. This article reviews the current molecular perspectives and diagnosis of oral cancer with proteomics using matrix-assisted laser desorption ionization (MALDI) and surface-enhanced laser desorption ionization (SELDI) mass spectrometry (MS). This method shows higher sensitivity, accuracy, reproducibility and ability to handle complex tissues and biological fluid samples. However, the data interpretation tools of contemporary mass spectrometry still warrant further improvement. Based on the data available with laser-based mass spectrometry, biomarkers of OSCC are classified as (i) diagnosis and prognosis, (ii) secretory, (iii) recurrence and metastasis, and (iv) drug targets. Majority of these biomarkers are involved in cell homeostasis and are either physiologic responders or enzymes. Therefore, proteins directly related to tumorigenesis have more diagnostic value. Salivary secretory markers are another group that offers a favourable and easy strategy for non-invasive screening and early diagnosis in oral cancer. Key molecular inter-related pathways in oral carcinogenesis are also intensely researched with software analysis to facilitate targeted drug therapeutics. The review suggested the need for incorporating 'multiple MS or tandem approaches' and focusing on a 'group of biomarkers' instead of single protein entities, for making early diagnosis and treatment for oral cancer a reality.

Contact Endoscopy of mucosal lesions of oral cavity - Preliminary experience.

BACKGROUND: Contact Endoscopy is a non invasive tool to visualise alterations in cell architecture in vivo. In this study we investigated the diagnostic accuracy of Contact Endoscopy in detecting malignancy in oral mucosal lesions. METHODS: 76 patients with oral mucosal lesions requiring biopsy were included. Contact Endoscopy was performed by Otolaryngologist before biopsy and findings recorded. The lesion was then biopsied and sent for histopathological examination by Pathologist who was blinded to Contact Endoscopy findings. Findings of Contact Endoscopy were compared with histopathological findings taking the latter as the gold standard. Two biopsies were reported as 'inconclusive' on histopathological examination and hence excluded from the final analysis. Data of the remaining 74 patients is presented here. RESULTS: Clinically lesions were diagnosed as ulcero-proliferative lesions in 34 patients, Leukoplakia in 19, Erythroplakia in 9, Lichen planus in 5 and Submucous fibrosis in 7 patients. Histopathological examination revealed presence of malignancy in 97.06% of ulcero-proliferative mucosal lesions, 10.53% of leukoplakia and 33.33% of erythroplakia while corresponding figures on Contact Endoscopy were 94.12%, 5.26% and 11.11% respectively. No malignancy was detected in lichen planus and submucous fibrosis by either technique. When compared with histopathological examination, CE showed sensitivity of 84.21%, specificity of 94.44% and accuracy of 89.19%. No adverse effects on the patients were seen due to the procedure or stain. CONCLUSION: Contact Endoscopy may be useful in determining cellular structure in vivo without biopsy to detect oral malignancy early. Further studies are suggested.

A Study of the Newer Prognostic Markers in Diffuse Large B Cell Lymphomas.

INTRODUCTION: Diffuse large B cell lymphomas (DLBCL) encompass a pathogenetically heterogeneous group of aggressive tumours that are rapidly fatal if untreated. Gene expression profiling studies have identified distinct molecular subtypes of DLBCL, one with an expression profile similar to normal germinal centre B cells (GCB subtype) and a second mimicking activated peripheral blood B cells (ABC subtype) having different prognostic significance allowing risk stratification of lymphoma patients and development of specific therapeutic strategies. METHODS: Twenty cases of DLBCL were included in the study and categorized into germinal centre and non germinal centre subtypes using the following antibody panel. CD10, Bcl-6, MUM1 and CD138. The germinal centre and non germinal centre subtypes were defined as under Germinal centre (DLBCL) CD10 + and/or Bcl-6 +, MUM1 -, CD138 - and Non germinal centre (DLBCL) CD10, Bcl-6 ±, MUM1 ±, CD138 ±. RESULT: In our study of twenty cases thirteen were germinal centre DLBCL while seven of the twenty cases were non germinal centre type of DLBCL. 75% of the nodal cases and 62.5% of extra nodal cases were germinal centre B cell type. Overall survival in the GCB and non GCB groups was 91% and 14% respectively and the difference was highly significant statistically. CONCLUSION: This study validates the existence of prognostic subgroups of DLBCL in the Indian population.

Pulmonary alveolar microlithiasis: Incidental finding - should we Ignore?

Pulmonary alveolar microlithiasis (PAM) is a rare entity, presenting mostly as an incidental finding. This disease has an autosomal recessive inheritance with inactivating mutations in the gene "solute carrier family 34 member 2". The present study was conducted to bring attention to this rare though preventable disease. The study was a cross-sectional descriptive study, conducted at the Department of Pathology, of a tertiary care hospital in New Dehli-India. PAMs were incidentally seen in two patients diagnosed with micronodular hepatic cirrhosis leading to reanalysis of 212 autopsies, retrospectively. Statistical analysis was done using Stata 14.0. We observed three forms (Type A, B and C) of round hyaline bodies measuring in diameter with thin delicate, radiating fibrils. These bodies were PAS positive, showed black discolouration of the pigment with von Kossa stain and birefringence on polarized microscopy using Congo red stain, however the refringence was light green as compared to apple green birefringence seen with amyloid deposition. PAM has a slow progressive course leading to a high rate of incidental detection. Drugs known to inhibit the micro-crystal growth of hydroxyapatite may slow the disease progression. The family members of patients with PAM may also be kept on follow up with regular imaging. Key messages: It is important to bring out the incidental finding as, seemingly innocuous observations may provide valuable insight into incurable diseases, especially rare diseases.