An inexpensive and rapid diagnostic method for detection of SARS-CoV-2 RNA by loop-mediated isothermal amplification (LAMP).

SARS-CoV-2 is a public pandemic health concern globally. Nasopharyngeal and oropharyngeal swab samples are used for Covid-19 viral detection. Sample collection procedure was tedious and uncomfortable and unsuitable for biochemical and CBC analysis in swab samples. Biochemistry and CBC tests are key determinant in management of Covid-19 patients. We developed a LAMP test to detect viral RNA in blood samples. LAMP is required four specific primers targeting the internal transcribed S-region and loop primers for viral RNA amplification. RNA was extracted from blood samples by TRIzol method. LAMP reaction was performed at 60 °C for 1 hour and amplicons were visualized in HNB dye. No cross-reactivity was seen with HBV, HCV, and HIV infected sample. Out of 40 blood samples, 33 samples were positive for LAMP and Q-PCR analysis, one sample was positive for LAMP and negative for Q-PCR, two samples were negative for LAMP but positive for Q-PCR, and four blood samples were negative for LAMP and Q-PCR. LAMP method has an accuracy of 92.50%, with sensitivity and specificity of 94.28% and 80%, respectively. Thus, LAMP diagnostic test has proved reliable, fast, inexpensive and can be useful for detection where the limited resources available.•LAMP method is a potential tool for detection of SARS-CoV-2.•Blood samples are the key determinant for routine diagnostics as well as molecular diagnostics.•LAMP assay is an appropriate diagnostics method which offers greater simplicity, low cost, sensitivity, and specificity than other methods in molecular diagnostics.

Impact of Yoga Intervention in Lower Limb Amputees following Trauma in Relation to Behavior and Quality of Life: A Randomized Controlled Trial.

Background: Yoga is an emerging intervention causing improvement in physical, mental, and spiritual well-being. Its role in improving outcomes in patients with amputation was investigated. Methodology: Patients with traumatic lower limb amputation (n = 50) were enrolled and randomized to the Yoga group (n = 26) against control (n = 24). Sociodemographic details, quality of life (QOL) (World Health Organization QOL-BREF), Depression anxiety stress scale (DASS), Rosenberg self-esteem scale, and amputee body image score were applied at baseline, 6,-18 weeks of amputation. Results: Sociodemographic and clinical variables were comparable between groups. At 18 weeks, the Yoga group had better QOL (P = 0.005) than the control group. Symptoms of depression (0.02) and anxiety (<0.001) reduced, and self-image (P = 0.015) improved significantly at 6 weeks, while stress (P = 0.003) reduced at 18 weeks in the yoga group. Despite comparable body image scores, the prosthesis usage (hours/day) was more (P = 0.005) in the Yoga (6.9 ± 3.2) group against the control (12 ± 2.7). Conclusion: Yoga improves QOL and self-esteem and reduces depression, anxiety, and stress symptoms in patients with traumatic amputation.

Filariasis in an alcoholic male-coincidental or causal association?

Lymphatic filariasis is a parasitic disease transmitted by mosquitoes that carry minute, thread-like worms. There are usually no signs or symptoms to be seen in patients with this disease for the most part. Some people, however, develop a condition called elephantiasis, which is marked by severe swelling in the arms, legs, breasts, or genitals. We here descibe an interesting case of a 40-year-old alcoholic brought to the emergency department with an alleged history of injury and incidentally found to have microfilariasis after a thorough blood smear screening. This case illustrates the need for smear evaluation in diagnosing microfilariasis and also begs whether these patients may be more susceptible to parasitic infections due to their impaired immune system.

Computation and Collation of Torque Expression in 0.018 Inch and 0.022 Inch Preadjusted Bracket Slots on Passive Insertion of Full-Size Archwire: A Finite Element Study.

Aim: To quantify and compare the prescribed torque expressed in preadjusted edgewise 0.018-inch and 0.022-inch bracket slot on passive insertion of full-size archwire using finite element method. Materials and Methods: Geometric model of the maxillary arch created with dimensions of the brackets and archwires scanned using microcomputed tomography were converted to finite element model using HYPERMESH V 12.0 software. Material properties were assigned and boundary conditioning was given. 0.017 × 0.025 inch, 0.018 × 0.025 inch, 0.019 × 0.025 inch, 0.021 × 0.025 inch stainless steel wires were inserted and ligated using 0.010 inch stainless steel wire. The amount of torque in each wire was quantified and compared. Results: 0.018 × 0.025-inch stainless steel archwire in 0.018-inch slot expressed more torque than 0.017 × 0.025-inch wire; in 0.022-inch slot 0.021 × 0.025-inch wire showed more torque. Torque expression in 0.018-inch slot was more than 0.022-inch slot. Torque expressed was less than the prescribed value in both bracket slots.

ACE2 protein expression in lung tissues of severe COVID-19 infection.

Angiotensin-converting enzyme 2 (ACE2) is a key host protein by which severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) enters and multiplies within cells. The level of ACE2 expression in the lung is hypothesised to correlate with an increased risk of severe infection and complications in COrona VIrus Disease 2019 (COVID-19). To test this hypothesis, we compared the protein expression status of ACE2 by immunohistochemistry (IHC) in post-mortem lung samples of patients who died of severe COVID-19 and lung samples obtained from non-COVID-19 patients for other indications. IHC for CD61 and CD163 was performed for the assessment of platelet-rich microthrombi and macrophages, respectively. IHC for SARS-CoV-2 viral antigen was also performed. In a total of 55, 44 COVID-19 post-mortem lung samples were tested for ACE2, 36 for CD163, and 26 for CD61, compared to 15 non-covid 19 control lung sections. Quantification of immunostaining, random sampling, and correlation analysis were used to substantiate the morphologic findings. Our results show that ACE2 protein expression was significantly higher in COVID-19 post-mortem lung tissues than in controls, regardless of sample size. Histomorphology in COVID-19 lungs showed diffuse alveolar damage (DAD), acute bronchopneumonia, and acute lung injury with SARS-CoV-2 viral protein detected in a subset of cases. ACE2 expression levels were positively correlated with increased expression levels of CD61 and CD163. In conclusion, our results show significantly higher ACE2 protein expression in severe COVID-19 disease, correlating with increased macrophage infiltration and microthrombi, suggesting a pathobiological role in disease severity.

Clinico-pathological features in fatal COVID-19 infection: a preliminary experience of a tertiary care center in North India using postmortem minimally invasive tissue sampling.

OBJECTIVES: To study the histopathology of patients dying of COVID-19 using post-mortem minimally invasive sampling techniques. METHODS: This was a single-center observational study conducted at JPNATC, AIIMS. Thirty-seven patients who died of COVID-19 were enrolled. Post-mortem percutaneous biopsies were taken from lung, heart, liver, kidney and stained with hematoxylin and eosin. Immunohistochemistry was performed using CD61 and CD163. SARS-CoV-2 virus was detected using IHC with primary antibodies. RESULTS: The mean age was 48.7 years and 59.5% were males. Lung histopathology showed diffuse alveolar damage in 78% patients. Associated bronchopneumonia was seen in 37.5% and scattered microthrombi in 21% patients. Immunopositivity for SARS-CoV-2 was observed in Type II pneumocytes. Acute tubular injury with epithelial vacuolization was seen in 46% of renal biopsies. Seventy-one percent of liver biopsies showed Kupffer cell hyperplasia and 27.5% showed submassive hepatic necrosis. CONCLUSIONS: Predominant finding was diffuse alveolar damage with demonstration of SARS-CoV-2 protein in the acute phase. Microvascular thrombi were rarely identified in any organ. Substantial hepatocyte necrosis, Kupffer cell hypertrophy, microvesicular, and macrovesicular steatosis unrelated to microvascular thrombi suggested that liver might be a primary target of COVID-19.

Anaplastic lymphoma kinase immunohistochemistry in lung adenocarcinomas: Evaluation of performance of standard manual method using D5F3 antibody.

OBJECTIVE: Immunohistochemistry (IHC) with anaplastic lymphoma kinase (ALK) antibodies is considered as an economical screening method in lung adenocarcinomas. Automated Ventana D5F3-IHC is approved by US Food and Drug Administration for targeted therapy; however, the automated IHC apparatus are not widely used in most laboratories. We evaluated the performance of ALK IHC using the manual semiquantitative method to assess the concordance with Ventana ALK IHC assay. MATERIALS AND METHODS: We tested 156 cases of primary lung adenocarcinomas for ALK protein expression by D5F3-IHC. The intensity of cytoplasmic staining was classified as 0 or 1+/2+/3+ (weak/medium/strong). Binary score of positive and negative was used for Ventana assay. A comparison analysis and clinicopathological features were recorded. RESULTS: ALK IHC was positive in 25 (16.02%) cases, of which 18 were men and mostly nonsmokers. The mean age for all patients was 55 years, and for ALK IHC-positive cases was 48 years. Nine of 25 (36%) ALK IHC-positive cases showed signet ring cell and mucinous morphology. On comparison, all, but one, cases positive by manual method showed positive results by automated assay. IHC negative cases by manual method were negative by Ventana assay. CONCLUSION: Manual IHC is equally effective in the detection of ALK-rearranged cases as automated methods. It can be easily integrated as a screening method into routine practice thus reducing the cost of automated systems. However, equivocal cases should be tested by approved methods.

Comparative evaluation of two flowcytometric analysers as diagnostic tools for the automated detection of malaria.

GOALS: Automated flowcytometric analysers are used to analyse complete blood count (CBC) in most patients. They differentiate WBC depending on their size, internal granularity, and nuclear content by producing various scattergrams. In malaria, haemozoin pigment-containing cells (parasitized RBC and phagocytic cells) are able to depolarize the laser beam and therefore exhibit abnormal scattergram. PROCEDURE: A study was conducted to evaluate the diagnostic utility and to determine the sensitivity and specificity of flowcytometric haematology analysers [Sysmex XE-2100 (System 1) and Cell-Dyn 3700 (System 2)] in comparison with the conventional microscopy (QBC, AO, Giemsa) and ICT (Immunochromatographic test) for detection of the malaria parasite. RESULTS: Fifty-eight of 320 samples were found to be positive for malaria by conventional methods. Considering atypical scattergram, System 1 showed the sensitivity and specificity of 68.9% and 90.6%, respectively; whereas System 2 showed a sensitivity of 100% and specificity of 98.8%. CONCLUSION: System 2 (Cell-Dyn 3700) was found to be a highly sensitive and specific tool compared to System 1 and conventional methods. Hence, it may be preferred for automated detection of malaria in the blood samples of patients with a differential diagnosis of malaria and those who are subjected to CBC analysis.

Experiments and ANFIS modelling for the biodegradation of penicillin-G wastewater using anaerobic hybrid reactor.

The performance of an anaerobic hybrid reactor (AHR) for treating penicillin-G wastewater was investigated at the ambient temperatures of 30-35°C for 245 days in three phases. The experimental data were analysed by adopting an adaptive network-based fuzzy inference system (ANFIS) model, which combines the merits of both fuzzy systems and neural network technology. The statistical quality of the ANFIS model was significant due to its high correlation coefficient R(2) between experimental and simulated COD values. The R(2) was found to be 0.9718, 0.9268 and 0.9796 for the I, II and III phases, respectively. Furthermore, one to one correlation among the simulated and observed values was also observed. The results showed the proposed ANFIS model was well performed in predicting the performance of AHR.

Automated detection of malaria with haematology analyzer Sysmex XE-2100.

BACKGROUND: Diagnosis of malaria is usually made by microscopy [Giemsa, Acridine Orange (AO), and Quantitative Buffy Coat (QBC) assay], which requires expertise. Currently, automated haematology analyzers are being used for complete blood count (CBC), in all acute febrile and non-febrile illnesses which simultaneously detects malaria. The normal scattergram by the analyzer (Sysmex 2100) comprises of five parameters i.e. lymphocytes (pink), monocytes (green), neutrophils (blue), eosinophils (red) with a space between the neutrophil and eosinophil populations. AIMS: We carried out a prospective study to compare the efficacy of Sysmex XE-2100 (Sysmex Corporation, Kobe) for detection of malaria in comparison to other conventional techniques. MATERIALS AND METHODS: 430 cases were analyzed for malaria by microscopy (QBC, AO, Giemsa), ICT (Immunochromatography) and flowcytometric analyzer (Sysmex XE-2100). The abnormal scattergrams were observed as double neutrophil, double eosinophil, grey zone, extended neutrophil zone with a decrease space between eosinophil and neutrophil, and a combination of above patterns. RESULTS: Out of 70 positive cases [49/70 (70%) P. vivax, 18/70 (25.7%) P. falciparum, and 3/70 (4.2%) both P. vivax and P. falciparum], 52 showed abnormal scattergrams by the analyzer. The sensitivity and specificity of hematology analyzer found to be 74.2% and 88%, respectively. CONCLUSION: Flowcytometric analyzer is a rapid, high throughput device which needs less expertization for the diagnosis of malaria. Hence, it can be used in the diagnostic laboratories as an early modality for diagnosis of malaria in suspected as well as clinically in apparent cases.

Blood transfusion practices at a level one trauma centre: a one-year retrospective review.

INTRODUCTION: Knowledge of blood usage patterns helps to address major issues such as the management of massive transfusion events, minimisation of transfusion risks, as well as in dealing with blood shortages. The aim of our study was to audit blood component usage at a Level I trauma centre blood bank. METHODS: A retrospective analysis of the transfusion data of 4,320 patients who were admitted to the General Surgery, Neurosurgery, Orthopaedics and Emergency Medicine departments during a one-year period was conducted. RESULTS: A total of 4,054 patients underwent transfusion. 88 percent, 94 percent, 80 percent and 100 percent of patients admitted to the General Surgery, Orthopaedics, Neurosurgery and Emergency Medicine departments, respectively, received transfusions. Packed cells were the most commonly utilised component, followed by fresh frozen plasma (FFP) and platelets in the ratio 3:2:1. The highest number of FFPs (2,052 units) and platelet concentrates (950 units) were used in the General Surgery and Neurosurgery departments, respectively. The calculated cross-match to transfusion (C:T) ratio did not exceed 2.5 in any of the departments. Among those transfused, the massive blood transfusion rate was low (1.77 percent). The rates of transfusion reactions and non-group-specific transfusions were also low (0.42 percent and 0.07 percent, respectively). CONCLUSION: The rate of transfusion of trauma patients was high (94 percent). Using the C:T ratio as a marker, optimal blood utilisation was noted in all departments. The methods of reporting transfusion reactions need to be revised. Future studies on the appropriateness of blood use and blood ordering schedules are required.

Nevoid basal cell carcinoma syndrome (Gorlin's syndrome): a case report.

A case of nevoid basal cell carcinoma syndrome is presented and its varied clinical manifestations and multi-system involvement are emphasised. Our case presented with an early onset of symptoms but sought medical help later on for progressively increasing jaw swelling and pain. On further evaluation, multiple pigmented skin papules, palmar pits, multiple jaw cysts, skull bone osteoporosis, bifid ribs and kyphosis were present. Systemic involvement was minimal. There was no significant family history.

Fine needle aspiration of tophi in asymptomatic gout--a case report.

Gout, a chronic hyperuricemic crystal induced arthropathy, may produce soft tissue masses (tophi). Tophi may be found in synovial membranes, periarticular ligaments, tendons, soft tissues as well as internal organs. We present a case in which diagnosis of gout was made by fine needle aspiration of tophus. The patient had a painless nodule over right ankle which was progressively increasing in size. He gave a past history of painful arthropathy, but serum uric acid levels were within normal limits. At this juncture, FNAC of the ankle tophus was performed which revealed aggregated and innumerable dissociated needle-shaped negatively birefringent crystals of monosodium urate (MSU) on polarization microscopy.

Pancytopenia--a clinico haematological study of 200 cases.

The present study was designed to ascertain the percentage of occurrence and causes of pancytopenia. All the cases of pancytopenia from July 2001 to June 2002 (one year) were examined in the Department of haematology, Safdarjung Hospital, New Delhi. Bone marrow aspirations/biopsy were performed in most of the cases (200 out of 250 cases). The commonest cause of pancytopenia, in our hospital was Megaloblastic anaemia (72%), followed by Aplastic anaemia (14%) and others.