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RT-PCR is the gold standard for diagnosis of COVID-19. All RT-PCR kits are based on RNA extraction from the clinical sample. There was a sudden increase in demand of these kits, both RNA extraction and COVID-19 RT-PCR kits during the pandemic. This sudden spurt in global demand created a situation of shortage of consumables, especially the RNA extraction kits. Hence, this study was carried out to evaluate and compare COVID-19 RT-PCR without RNA extraction step using buffer R3. Sensitivity, specificity and accuracy of RT-PCR kit without RNA extraction were 89.16 %, 100% and 89.6% respectively. This approach saved more than 50 % time compared to the RT-PCR kit with RNA extraction approach allowing enhanced daily sample processing capability. RT-PCR kit without RNA extraction help in managing a greater number of samples, reduces cost and turnaround time.
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COVID-19 , ARN Viral , Reacción en Cadena en Tiempo Real de la Polimerasa , SARS-CoV-2 , Sensibilidad y Especificidad , Humanos , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación , COVID-19/diagnóstico , COVID-19/virología , ARN Viral/genética , ARN Viral/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Juego de Reactivos para Diagnóstico/normas , Prueba de Ácido Nucleico para COVID-19/métodos , Manejo de Especímenes/métodosRESUMEN
Many drugs were recommended as antiviral agents for infection control and effective therapy to reduce the mortality rate for COVID-19 patients. Hydroxychloroquine (HCQ), an antimalarial drug, has been controversially recommended for prophylactic use in many countries, including India, to control SARS-CoV-2 infections. We have explored the effect of prophylactic HCQ from the cells of bronchoalveolar lavage fluids from COVID-19-induced acute respiratory distress syndrome patients to determine the level of infection and ultrastructural alterations in the ciliated epithelium, type II pneumocytes, alveolar macrophages, neutrophils, and enucleated granulocytes. Ultrastructural investigation of ciliated epithelium and type II pneumocytes showed lesser infections and cellular impairment in the prophylactic HCQ+ group than HCQ- group. However, macrophages and neutrophils displayed similar infection and ultrastructural alterations in both patient groups. The enucleated fragments of granulocytes showed phagocytosis of the matured virus in HCQ+ groups. The present report unveils the ultrastructural proof to complement the paradox regarding the role of prophylactic HCQ in COVID-19 patients.
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COVID-19 , Humanos , Hidroxicloroquina/efectos adversos , SARS-CoV-2 , Líquido del Lavado Bronquioalveolar , Tratamiento Farmacológico de COVID-19 , Antivirales/efectos adversosRESUMEN
AIM: To find out the relative incidence and outcome of posterior fossa abnormality (PFA) in terms of survival at birth until 2 years of age. METHODS: We conducted a prospective study; all fetuses diagnosed with posterior fossa abnormality were followed-up. The outcome was observed with respect to survival, the presence of associated anomalies, the existence of developmental delay after a telephonic interview. RESULTS: Out of 2703 children with congenital anomalies, 921 (34.1%) had a central nervous system defect; 76 cases of PFA were fully followed. Dandy-Walker malformation (DWM) was present in 50% (38/76), mega cisterna magna 18.4% (14/76), Blake pouch cyst 13.2% (10/76), vermian hypoplasia (VH) 13.2% (10/76) and arachnoid cyst 5.2% (4/76). The diagnosis was possible before 20 weeks in only 12 (15.8%) cases. The mean gestational age at delivery was 34.7 ± 6.7 weeks. Associated anomalies were seen in 35/76 (46.1%) cases. A total of 35/76 (46.1%) survived after 2 years; there was developmental delay in 9.2% of cases. CONCLUSION: There is a large variation in the outcome of PFA depending upon the type of anomaly. Associated anomalies are common in VH and DWM, making their prognosis worse.
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Quistes , Síndrome de Dandy-Walker , Femenino , Recién Nacido , Niño , Humanos , Lactante , Prevalencia , Estudios Prospectivos , Imagen por Resonancia Magnética/métodos , Síndrome de Dandy-Walker/epidemiología , Síndrome de Dandy-Walker/diagnóstico , Síndrome de Dandy-Walker/genética , FetoRESUMEN
In this study, we examined the cellular infectivity and ultrastructural changes due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in the various cells of bronchoalveolar fluid (BALF) from intubated patients of different age groups (≥60 years and <60 years) and with common comorbidities such as diabetes, liver and kidney diseases, and malignancies. BALF of 79 patients (38 cases >60 and 41 cases <60 years) were studied by light microscopy, immunofluorescence, scanning, and transmission electron microscopy to evaluate the ultrastructural changes in the ciliated epithelium, type II pneumocytes, macrophages, neutrophils, eosinophils, lymphocytes, and anucleated granulocytes. This study demonstrated relatively a greater infection and better preservation of subcellular structures in these cells from BALF of younger patients (<60 years compared with the older patients (≥60 years). The different cells of BALF from the patients without comorbidities showed higher viral load compared with the patients with comorbidities. Diabetic patients showed maximum ultrastructural damage in BALF cells in the comorbid group. This study highlights the comparative effect of SARS-CoV-2 infection on the different airway and inflammatory cells of BALF at the subcellular levels among older and younger patients and in patients with comorbid conditions.
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COVID-19 (Coronavirus Disease 2019), illness with associated comorbidities and corticosteroid therapy makes the host immunocompromised and prone to opportunistic microbial infections. As the world continues to struggle with the pandemic of COVID-19, an increase in cases of opportunistic fungal infections have been reported from all over the world during the second wave of COVID-19 like aspergillosis, mucormycosis, and candidiasis. Scedosporium apiospermum is an emerging pathogen that is usually associated with mycetoma, pulmonary infection, and central nervous infections. It has been rarely associated with fungal rhinosinusitis (FRS). In this study, a rare case of FRS caused by S.apiospermum in an immunocompromised post-Covid-19 diabetic woman is reported.
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Whiteflies are a group of universally occurring insects that are considered to be a serious pest in their own way for causing both direct and indirect damages to crops. A few of them serve as vectors of plant viruses that are detrimental to the crop in question and cause an actual loss in productivity. A lot of attention is focused on pest control measures under the umbrella of IPM. In this review, we attempt to summarize the existing literature on how and why whiteflies are a serious concern for agriculture and society. We reviewed why there could be a need for fresh insight into the ways and means with which the pest can be combated. Here, we have emphasized next-generation strategies based on macromolecules, i.e., RNA interference and genetic engineering (for the expression of anti-whitefly proteins), as these strategies possess the greatest scope for research and improvement in the future. Recent scientific efforts based on nanotechnology and genome editing, which seem to offer great potential for whitefly/crop pest control, have been discussed. Comprehensive apprehensions related to obstacles in the path of taking lab-ready technologies into the farmers' field have also been highlighted. Although the use of RNAi, GM crops, nanotechnologies, for the control of whiteflies needs to be evaluated in the field, there is an emerging range of possible applications with promising prospects for the control of these tiny flies that are mighty pests.
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BACKGROUND: Synechococcus sp. PCC 7002 is an attractive organism as a feedstock and for photoautotrophic production of biofuels and biochemicals due to its fast growth and ability to grow in marine/brackish medium. Previous studies suggest that the growth of this organism is limited by the HCO3 - transport across the cytoplasmic membrane. Tools for genetic engineering are well established for this cyanobacterium, which makes it possible to overexpress genes of interest. RESULTS: In this work, we overexpressed two different native Na+-dependent carbon transporters viz., SbtA and BicA in Synechococcus sp. PCC 7002 cells under the influence of a strong light-inducible promoter and a strong RBS sequence. The overexpression of these transporters enhanced biomass by about 50%, increased intracellular glycogen about 50%, and increased extracellular carbohydrate up to threefold. Importantly, the biomass and glycogen productivity of the transformants with air bubbling was even higher than that of WT cells with 1% CO2 bubbling. The overexpression of these transporters was associated with an increased carotenoid content without altering the chl a content. CONCLUSIONS: Our work shows the utility of increased carbon transport in improving the growth as well as product formation in a marine cyanobacterium and will serve to increase the utility of this organism as a potential cell factory.
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AIM: To study the spectrum of neurological complications of diphtheria, timing of onset with respect to respiratory disease, and pattern of recovery. SETTINGS AND DESIGN: Prospective, observational, hospital-based study conducted in tertiary care hospital. MATERIALS AND METHODS: Twenty-eight cases of diphtheria with neurological complications were admitted during the period of study. Demographic profile, age, gender, status of immunization, past history stressing on the severity of the respiratory disease, and complaints regarding diverse complications of diphtheria were recorded. Detailed clinical and central nervous system examinations along with relevant investigations were carried out. RESULTS: Children were in the age group of 3-18 years. All 28 children presented with bulbar symptoms. Isolated palatal palsy was present in 18 children (64%). Third cranial involvement was present in four children. Three children had unilateral lower motor neuron facial palsy and one child had sixth cranial nerve palsy. Nine children developed symmetric limb weakness. Diaphragmatic palsy was present in three children with the onset from 1-3 weeks after pharyngeal diphtheria. Loss of vasomotor tone was present in two children. Recovery was complete in all 28 children. CONCLUSION: Pediatricians/neurophysicians should have a high index of suspicion to recognize diphtheritic polyneuropathy. It carries good prognosis, hence timely diagnosis and differentiation from other neuropathies is a prerequisite for rational management and contact tracing.
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A seven-year-old boy with nephrotic syndrome presented with a frequent rash along with relapse of nephrotic syndrome. Clinical and histological features were suggestive of pityriasis lichenoides et varioliformis acuta (PLEVA). Treatment of the condition with doxycycline led to the cure of the lesions as well as the relapses.
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Síndrome Nefrótico/etiología , Pitiriasis Liquenoide/complicaciones , Antibacterianos/uso terapéutico , Niño , Doxiciclina/uso terapéutico , Humanos , Masculino , Pitiriasis Liquenoide/tratamiento farmacológico , Recurrencia , Resultado del TratamientoRESUMEN
The Rhesus G antigen is present on all RBCs that are C+ and also on most D+ RBCs. Due to this co-distribution of G with either C or D antigen, it mimics a reactivity pattern of anti C + anti D on Indirect Antiglobulin Test (IAT), though the role of Anti G in causing Hemolytic Disease of Newborn (HDN) is controversial. The differentiation of anti D, anti C, and anti G is essential particularly in pregnant females. We hereby report a rare case of anti G with anti D and anti C in a pregnant woman with emphasis on approach to identify anti D+C+G and its implications.
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Immune hemolytic anemia is characterized by clinical and laboratory features of hemolytic anemia with direct antiglobulin test (DAT) positivity. It could be autoimmune hemolytic anemia (AIHA), alloimmune, or drug-induced hemolysis based on the antigenic stimulus. Furthermore, based on thermal amplitude of autoantibody, AIHA is classified as warm (65%), cold (30%), and mixed (5%) type. Mixed AIHA is extremely rare in children and must be differentiated from warm AIHA with clinically insignificant cold agglutinins and cold hemagglutinin disease as their treatment is different. It may present as blood group discrepancy or cross-match incompatibility leading to delay in arranging suitable blood unit for transfusion. Therefore, a thorough immunohematology workup including monospecific DAT, indirect antiglobulin test at 4°C and 37°C, determination of thermal amplitude and titer is essential. We hereby present two pediatric cases of mixed AIHA presenting as ABO forward and reverse blood group discrepancy and cross-match incompatibility.
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Whitefly (Bemisia tabaci) damages field crops by sucking sap and transmitting viral diseases. None of the insecticidal proteins used in genetically modified (GM) crop plants to date are effective against whitefly. We report the identification of a protein (Tma12) from an edible fern, Tectaria macrodonta (Fee) C. Chr., that is insecticidal to whitefly (median lethal concentration = 1.49 µg/ml in in vitro feeding assays) and interferes with its life cycle at sublethal doses. Transgenic cotton lines that express Tma12 at â¼0.01% of total soluble leaf protein were resistant to whitefly infestation in contained field trials, with no detectable yield penalty. The transgenic cotton lines were also protected from whitefly-borne cotton leaf curl viral disease. Rats fed Tma12 showed no detectable histological or biochemical changes, and this, together with the predicted absence of allergenic domains in Tma12, indicates that Tma12 might be well suited for deployment in GM crops to control whitefly and the viruses it carries.
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Helechos/metabolismo , Gossypium/genética , Gossypium/parasitología , Hemípteros/virología , Insecticidas/metabolismo , Proteínas de Plantas/metabolismo , Animales , Helechos/genética , Mejoramiento Genético/métodos , Gossypium/virología , Hemípteros/patogenicidad , Proteínas de Plantas/genética , Ingeniería de Proteínas/métodos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMEN
Ectopic breast carcinoma in the axillary region is rare with an incidence ranging from 0.3-6%. We report a case of infiltrating duct carcinoma in an adult female arising in aberrant breast tissue in the axilla diagnosed on fine needle aspiration cytology. There was history of recent increase in size of the lump which was otherwise present for the past 5 years. This case highlights the role fine needle aspiration cytology can play in the early diagnosis of malignant transformation of lumps.
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Axila/patología , Enfermedades de la Mama , Neoplasias de la Mama , Mama , Carcinoma Ductal de Mama , Coristoma/diagnóstico , Disección/métodos , Ganglios Linfáticos , Pezones/anomalías , Adulto , Antineoplásicos/uso terapéutico , Biopsia con Aguja Fina/métodos , Enfermedades de la Mama/diagnóstico , Enfermedades de la Mama/patología , Neoplasias de la Mama/diagnóstico , Neoplasias de la Mama/patología , Neoplasias de la Mama/cirugía , Carcinoma Ductal de Mama/diagnóstico , Carcinoma Ductal de Mama/patología , Carcinoma Ductal de Mama/cirugía , Transformación Celular Neoplásica , Diagnóstico Diferencial , Detección Precoz del Cáncer , Femenino , Humanos , Ganglios Linfáticos/patología , Ganglios Linfáticos/cirugía , Pezones/patología , Resultado del TratamientoRESUMEN
Although malaria in pregnancy, lactation and congenital malaria can be a disease burden in the endemic zones of Africa and Indian sub-continent, it is still epidemiologically less investigated in India. As it may lead to considerable maternal and perinatal morbidity and mortality, awareness and timely intervention is necessary for desirable outcome and prevention of the condition. Very few reports of congenital malaria are available in the literature from an endemic country like India. Herein we describe a case of congenital malaria from north India in a 21-day neonate. Clinical presentation of this condition in the neonate may offer a considerable diagnostic challenge, and differentiation from vector borne malaria in infants may be important from the management point of view. Hence a review of the differential diagnosis, management and prevention of congenital malaria has been attempted in the Indian perspective.
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Programas Nacionales de Salud , Servicios de Salud Rural/estadística & datos numéricos , Cobertura Universal del Seguro de Salud/organización & administración , Estudios Transversales , Atención a la Salud/estadística & datos numéricos , Humanos , India , Programas Nacionales de Salud/normas , Programas Nacionales de Salud/estadística & datos numéricos , Evaluación de NecesidadesRESUMEN
A lectin was purified from the leaves of Allium altaicum and corresponding gene was cloned. The lectin namely Allium altaicum agglutinin (AAA) was ~24 kDa homodimeric protein and similar to a typical garlic leaf lectin. It was synthesized as 177 amino acid residues pre-proprotein, which consisted of 28 and 43 amino acid long N and C-terminal signal peptides, respectively. The plant expressed this protein more in scapes and flowers in comparison to the bulbs and leaves. Hemagglutination activity (with rabbit erythrocytes) was 1,428 fold higher as compared to Allium sativum leaf agglutinin (ASAL) although, the insecticidal activity against cotton aphid (Aphis gossypii) was relatively low. Glycan array revealed that AAA had higher affinity towards GlcAb1-3Galb as compared to ASAL. Homology analysis showed 57-94% similarity with other Allium lectins. The mature protein was expressed in E. coli as a fusion with SUMO peptide in soluble and biologically active form. Recombinant protein retained high hemagglutination activity.
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Allium/genética , Hemaglutininas/química , Lectinas de Plantas/química , Lectinas de Plantas/farmacología , Allium/química , Secuencia de Aminoácidos , Animales , Áfidos/efectos de los fármacos , Secuencia de Bases , Secuencia de Carbohidratos , Clonación Molecular , Eritrocitos/efectos de los fármacos , Escherichia coli , Hemaglutinación/efectos de los fármacos , Hemaglutininas/genética , Hemaglutininas/metabolismo , Insecticidas/química , Insecticidas/metabolismo , Insecticidas/farmacología , Modelos Moleculares , Datos de Secuencia Molecular , Filogenia , Hojas de la Planta/química , Lectinas de Plantas/genética , Lectinas de Plantas/metabolismo , Polisacáridos/química , Polisacáridos/metabolismo , Conejos , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Proteínas Recombinantes de Fusión/farmacología , Proteína SUMO-1/química , Proteína SUMO-1/metabolismo , Alineación de Secuencia , Espectrometría de Masas en TándemRESUMEN
Over expression of lectin genes in E. coli often gives inclusion bodies that are solubilised to characterize lectins. We made N-terminal fusion of the Allium sativum leaf agglutinin (ASAL) with SUMO (small ubiquitin related modifier) peptide. The SUMO peptide allowed expression of the recombinant lectin in E. coli, predominantly in soluble form. The soluble fusion protein could be purified by immobilized metal affinity column (IMAC), followed by size exclusion chromatography. The SUMO protease failed to cleave the SUMO peptide from ASAL. This may be due to steric hindrance caused by the homodimer structure of the chimeric ASAL. Some properties like dimerization, haemagglutination and insecticidal properties of the recombinant SUMO-ASAL fusion protein were comparable to the plant derived native lectin. However, glycan array analysis revealed that the carbohydrate binding specificity of the recombinant SUMO-ASAL was altered. Further, the fusion protein was not toxic to E. coli (native ASAL exhibited toxicity). The recombinant lectin was more thermo-labile as compared to the native lectin. Three important findings of this study are: (1) sugar specificity of ASAL can be altered by amino-terminal fusion; (2) anti-E. coli activity of ASAL can be eliminated by N-terminal SUMO fusion and (3) SUMO-ASAL may be a preferred candidate insecticidal protein for the development of transgenic plants.