Correlation of serotype-specific strain in patients with dengue virus infection with neurological manifestations and its outcome.

INTRODUCTION: Neurological manifestation of dengue virus infection is a rare entity. Serotypes commonly associated with neurological manifestation are DENV-2 and DENV-3. We plan to detect the serotypes related to the neurological presentation in dengue infection and its correlation with different neurological complications and outcome. METHODS: In this case-control study, consecutive dengue cases with different neurological manifestations were enrolled along with age and sex-matched controls (dengue patients without neurological complication). Serotyping using RT-PCR of samples of cases and controls were done. Level of correlation was analyzed with various parameters and outcomes. RESULTS: In cases out of 33 samples, 6 sample serotypes were detected, which were composed of DENV-1 (n = 2) and DENV-2 (n = 4). In controls, DENV-1 (n = 5), DENV-2 (n = 6), and DENV-3 (n = 3) were detected. When statistically correlated, no significant association was found in cases and controls with dengue virus serotype. The frequency of serotype 2 was higher in hypokalemic paralysis cases than non-hypokalemic paralysis cases and the difference was significant (p < 0.05). The outcome was good (mRS < 3) in all the cases where serotypes were detected, but on statistical correlation, it was not found significant (p > 0.05). CONCLUSION: DENV-1 and DENV-2 are associated with neurological manifestation of dengue infection, which is different from the existing literature, where DENV-2 and DENV-3 are reported. The detection of DENV serotype will help in predicting and best management of neurological complication. The serotype 2 of dengue virus is more commonly associated with dengue-associated hypokalemic paralysis than other neurological complication (p < 0.05). There is no significant association of serotypes with outcome or mortality.

Wuhan to World: The COVID-19 Pandemic.

COVID-19 is a Severe Acute Respiratory Syndrome (SARS), caused by SARS-CoV-2, a novel virus which belongs to the family Coronaviridae. It was first reported in December 2019 in the Wuhan city of China and soon after, the virus and hence the disease got spread to the entire world. As of February 26, 2021, SARS-CoV-2 has infected ~112.20 million people and caused ~2.49 million deaths across the globe. Although the case fatality rate among SARS-CoV-2 patient is lower (~2.15%) than its earlier relatives, SARS-CoV (~9.5%) and MERS-CoV (~34.4%), the SARS-CoV-2 has been observed to be more infectious and caused higher morbidity and mortality worldwide. As of now, only the knowledge regarding potential transmission routes and the rapidly developed diagnostics has been guiding the world for managing the disease indicating an immediate need for a detailed understanding of the pathogen and the disease-biology. Over a very short period of time, researchers have generated a lot of information in unprecedented ways in the key areas, including viral entry into the host, dominant mutation, potential transmission routes, diagnostic targets and their detection assays, potential therapeutic targets and drug molecules for inhibiting viral entry and/or its replication in the host including cross-neutralizing antibodies and vaccine candidates that could help us to combat the ongoing COVID-19 pandemic. In the current review, we have summarized the available knowledge about the pathogen and the disease, COVID-19. We believe that this readily available knowledge base would serve as a valuable resource to the scientific and clinical community and may help in faster development of the solution to combat the disease.

Role of TfR2-Y250X and TfR1- rs3817672 Single Nucleotide Polymorphism on Pathophysiology of Iron Deficiency Anemia.

BACKGROUND: Transferrin receptor (TfR) is a carrier protein for transferrin. It is regulated in response to intracellular iron concentration and plays a role for the import of iron into the cell. The transferring receptor 2 (TFR2) gene showed homology to transferrin receptor 1 (TFR1) gene and encodes a transmembrane protein with a large extracellular domain, which is able to bind transferrin. Mutations in transferrin receptors (TfR2 and TfR1) may alter the pathophysiology of iron deficiency anemia. Alteration in genes encoding transferring receptor cause change in iron homeostatsis and provides a tool for investigating the excess iron absorption and abnormal iron distribution in iron related disorders. However the clinical significance of the interaction of transferring mutations with iron deficiency anemia remains unclear. Thus, the objective of my study was to investigate the effect of TFR1 and TFR2 genotypes on pathophysiology of iron deficiency anemia. STUDY DESIGN: Study subjects were 460 iron deficiency anemia patients and 500 age and sex-matched healthy controls. Transferrin receptor, ferritin and CRP analysis was done by ELISA method while ESR analysis was done according to Wintrobes's method. CBC analysis was done by auto-analyzer. TFR1-rs3817672 SNP and TFR2 (Y250X) mutation was analyzed by using PCR RFLP method. RESULTS: Amongst the iron deficiency anemia patients, 13 were heterozygous and five were homozygous for rs3817672 SNP. TFR2 (Y250X) mutation was detected in 6 patients with heterozygous conditions. None of the patients were presenting homozygous condition while four controls were presenting heterozygous and one with homozygous condition. Controls were presenting 3% and 0.6% of TFR1 rs3817672 SNP heterozygosity respectively. CONCLUSION: TfR2 -Y250X and TfR1-rs3817672 SNP showed clinical association with iron deficiency anemia and screening for mutations of TFR2 is a new diagnostic tool that can be offered to patients who do not have HFE mutations or who have incomplete HFE genotypes. This results may have practical implications for the molecular diagnosis of hemochromatosis. Genotyping the TFR gene should be included in the disease diagnostic protocols.

Simultaneous oxidation and esterification of cellulose for use in treatment of water containing Cu(II) ions.

Cellulose powder was modified with Sodium Hypochlorite (NaOCl) and Octenyl Succinic Anhydride (OSA) in different proportions. The adsorbents were characterised by BET, FTIR, XRD, TGA and SEM. Intrinsic viscosity increased with degree of substitution (DS). FTIR peaks confirmed the presence of carbonyl groups. Batch adsorption parameters studied were contact time, adsorbent dosage, pH and temperature. Adsorption process was influenced significantly by pH. Freundlich isotherm showed a better correlation coefficient (R2 and χ2) than Langmuir and Temkin isotherms. The adsorption process was inferred as physico-chemical adsorption process (based on isotherm and kinetic modelling). Pseudo second order kinetics model was found to fit well on the empirical data obtained. Adsorption of Cu(II) ions over the modified cellulose was found to be exothermic in nature (ΔH<0). HCl was found to be more effective desorbing agent than NaOH.

Study of Sleep Disorders and Polysomnographic Evaluation among Primary Chronic Daily Headache Patients.

OBJECTIVE: Studies related to sleep disorders and polysomnography (PSG) among chronic daily headache patients are rare. We studied this and compared chronic migraine (CM) with chronic tension-type headache. METHODS: Eighty-three patients were recruited. They were evaluated by semi-structured interview, headache, and sleep diaries along with Epworth Sleepiness Scale score and insomnia symptom score. Overnight PSG was performed and data compared. RESULTS: Chronic tension-type headache was more common than CM, both having female preponderance. Insomnia followed by excessive daytime sleepiness was prevalent sleep disorder. Sleep efficiency and Stage 3 sleep were lower in CM compared to chronic tension-type. ESSS was significantly increased among chronic tension-type patients. No significant correlation was found among PSG parameters in patients with or without sleep disorders. CONCLUSION: Insomnia being most common sleep disorder among chronic headache population. Chronic tension-type headache had slightly better slow-wave sleep than CM and significantly increased daytime sleepiness.

Phenotypic effect of α-globin gene numbers on Indian sickle ß-thalassemia patients.

BACKGROUND: Sickle cell ß-thalassemia is a compound heterozygous state of ß-thalassemia and sickle cell anemia. Patient with these conditions showed mild-to-severe clinical phenotype. OBJECTIVES: The objective of this study was to evaluate the effects of α-globin gene numbers on the phenotype of sickle cell ß-thalassemia patients. MATERIALS AND METHODS: Seventy-five sickle cell ß-thalassemia patients were characterized. Clinical, hematological, and molecular characterization was performed in all subjects. Amplified refectory mutation system-polymerase chain reaction was applied for ß-thalassemia mutation study while α-genotyping was conducted by Gap-PCR. RESULTS: Highest frequency of IVS1-5 (33 out of 75 patients) ß-thalassemia genotype was recorded. Twenty-eight patients were reported with α-globin chain deletion while four had α-triplications (Anti α-3.7kb). Sickle ß-thalassemia patients with α-chain deletions ameliorate hematological and clinical variables. CONCLUSIONS: This study indicates that the coexistence of α-globin chain deletions showed mild phenotype instead of absence of α-chain deletions while the patients with triplication of α-genes express severe phenotype.

Phenotypic heterogeneity of asian Indian inversion deletions gγ(aγδß)0 breakpoint a and breakpoint B.

Asian Indian inversion deletion Gγ (Aγδß)0-thalassemia is a rare entities characterized by high HbF. Due to interaction with various genetic factors, patients with Gγ (Aγδß)0-thalassemia showed clinical variability. Here we are presenting the phenotypic expression of Gγ(Aγδß)0 thalassemia under influence of various co-inherited factors. Patient with α-globin gene deletion had mild phenotype than the patient with ß-globin mutations. Patient with alpha gene deletion were presenting clinical character like thalassemia intermedia while Gγ (Aγδß)0-thalassemia patients with co- presence of beta thalssemia mutation clinically behaved like thalassemia major.

Molecular characterization of hemoglobin D Punjab traits and clinical-hematological profile of the patients.

CONTEXT AND OBJECTIVE: Hemoglobin (Hb) D hemoglobinopathies are widespread diseases in northwestern India and usually present with mild hemolytic anemia and mild to moderate splenomegaly. The heterozygous form of Hb D is clinically silent, but coinheritance of Hb D with Hb S or beta-thalassemia produces clinically significant conditions like thalassemia intermedia of moderate severity. Under heterozygous conditions with coinheritance of alpha and beta-thalassemia, patients show a degree of clinical variability. Thus, our aim was to molecularly characterize the Hb D trait among individuals who were clinically symptomatic because of co-inheritance of alpha deletions or any beta-globin gene mutations. DESIGN AND SETTING: This was a cross-sectional study conducted in an autonomous tertiary-care hospital. METHODS: Complete blood count and red cell indices were measured using an automated cell analyzer. Quantitative assessment of hemoglobin Hb F, Hb A, Hb A2 and Hb D was performed by means of high performance liquid chromatography (HPLC). DNA extraction was done using the phenol-chloroform method. Molecular analyses on common alpha deletions and common beta mutations were done using the Gap polymerase chain reaction and Amplification Refractory Mutation System, respectively. RESULTS: We evaluated 30 patients and found clinical variation in the behavior of Hb D traits. In six patients, the Hb D traits were clinically symptomatic and behaved like those of thalassemia intermedia. Molecular characterization showed that three out of these six were IVS-1-5 positive. CONCLUSIONS: HPLC may not be the gold standard for diagnosing symptomatic Hb D Punjab traits. Hence, standard confirmation should include molecular studies.

Effect of ANXA2 gene single nucleotide polymorphism (SNP) on the development of osteonecrosis in Indian sickle cell patient: a PCR-RFLP approach.

Osteonecrosis is a serious complication in sickle cell patients. The common sites of the necrosis are femoral head, head of the humerus and acetabulam. Annexin A2 (ANXA2) protein mainly functions in bone formation and bone resorption. Alteration of ANXA2 gene may affect the manifestations of osteonecrosis in the patients. PCR-RFLP is a common applicable technique for the detection of known mutation/polymorphisms. Here we are presenting application of the PCR-RFLP technique for determination of the ANXA2 gene single nucleotide polymorphism frequency and their clinical association among Indian sickle cell patients. Five known SNPs of ANXA2 gene (rs7170178, rs73435133, rs73418020, rs72746635 and rs73418025) were determined using the HpyCH4V, DdeI, HpyCH4III and Sau 961 restriction enzyme respectively. Restriction enzyme DdeI was common for rs73435133 and rs72746635 SNP. Only the rs7170178 SNP was detected among patient and control and the other four SNPs were absent in the studied groups. The frequency of ANXA2 gene rs7170178 SNP (A/G, G/G) was comparatively higher in sickle cell patients than controls and it was clinically associated with sickle cell osteonecrosis. The P value of heterozygotes (A/G) and homozygotes (G/G) genotypes were <0.001 and 0.001 respectively, which were highly significant. This study established the application of PCR-RFLP in detection of ANXA2 SNPs in sickle cell patients.

Modulating Effect of the -158 γ (C→T) Xmn1 Polymorphism in Indian Sickle Cell Patients.

Xmn1 polymorphism is a known factor, which increases fetal haemoglobin production. Among the inherited disorders of blood, thalassaemia and Sickle Cell Diseases contributes to a major bulk of genetic diseases in India. Our aim was to verify the role of the Xmn1 polymorphism as a modulating factor in sickle cell patients and frequency of the polymorphism in Indian sickle cell patients. 60 sickle homozygous and 75 sickle beta thalassemia patients were included and 5 ml blood sample was collected from them. Screening of sickle patients was done by HPLC. An automated cell analyzer SYSMEX (K-4500 Model) was used to analyze the Complete Blood Count of patients. Xmn1 polymorphism analysis was done by PCR-RFLP and one-way ANOVA test was applied to analysis of variance between groups. Among the sickle patients 27 were heterozygous (+/-) and 19 were homozygous (+/+) while 30 were heterozygous (+/-) and 24 were homozygous (+/+) in sickle ß-thalassemia patients. Extremely significant differences (p-value <0.001) of hematological parameters seen among patients with Xmn1 carrier and without the Xmn1 carrier. In our cases the clinical symptoms were barely visible and higher HbF level with Xmn1 carriers were found. Presence of Xmn1 polymorphism in sickle cell patients with higher HbF were phenotypically distinguished in the sickle cell patients. We conclude that the phenotypes of Indian sickle cell patients were greatly influenced by Xmn1polymorphism.

Prevalence of factor V Leiden G1691A, MTHFR C677T, and prothrombin G20210A among Asian Indian sickle cell patients.

The prevalence of factor V (FV) Leiden G1691A, prothrombin G20210A, and methylenetetrahydrofolate reductase (MTHFR) C677T mutations were investigated among 90 sickle trait, 61 sickle homozygous, 75 sickle beta thalassemia, and 15 HbSD Asian Indian sickle cell patients. In all, 297 healthy controls were evaluated to compare the polymorphism frequency. The prevalence of FV Leiden heterozygous G>A were significant in the group (P = .02), while PRT G20210A polymorphism was not seen among patients as well as controls. However, an increased frequency of the MTHFR 677 C>T genotype was seen among patients as well as controls, but this was not statistically significant (P = .13). This suggested a low impact of inherited hypercoagulability risk factors in the pathogenesis of sickle cell disease and/or its complications.

Molecular characterization of hemoglobin D Punjab traits and clinical-hematological profile of the patients / Caracterização molecular dos traços de hemoglobina D Punjab e perfil clínico-hematológico dos pacientes

CONTEXT AND OBJECTIVE: Hemoglobin (Hb) D hemoglobinopathies are widespread diseases in northwestern India and usually present with mild hemolytic anemia and mild to moderate splenomegaly. The heterozygous form of Hb D is clinically silent, but coinheritance of Hb D with Hb S or beta-thalassemia produces clinically significant conditions like thalassemia intermedia of moderate severity. Under heterozygous conditions with coinheritance of alpha and beta-thalassemia, patients show a degree of clinical variability. Thus, our aim was to molecularly characterize the Hb D trait among individuals who were clinically symptomatic because of co-inheritance of alpha deletions or any beta-globin gene mutations. DESIGN AND SETTING: This was a cross-sectional study conducted in an autonomous tertiary-care hospital. METHODS: Complete blood count and red cell indices were measured using an automated cell analyzer. Quantitative assessment of hemoglobin Hb F, Hb A, Hb A2 and Hb D was performed by means of high performance liquid chromatography (HPLC). DNA extraction was done using the phenol-chloroform method. Molecular analyses on common alpha deletions and common beta mutations were done using the Gap polymerase chain reaction and Amplification Refractory Mutation System, respectively. RESULTS: We evaluated 30 patients and found clinical variation in the behavior of Hb D traits. In six patients, the Hb D traits were clinically symptomatic and behaved like those of thalassemia intermedia. Molecular characterization showed that three out of these six were IVS-1-5 positive. CONCLUSIONS: HPLC may not be the gold standard for diagnosing symptomatic Hb D Punjab traits. Hence, standard confirmation should include molecular studies.

CONTEXTO E OBJETIVO: Hemoglobinopatias da hemoglobina (Hb) D são doenças amplamente disseminadas no noroeste da Índia e geralmente se apresentam com anemia hemolítica leve e esplenomegalia leve a moderada. A forma heterozigótica de Hb D é clinicamente silenciosa, mas co-herança de Hb D com Hb S ou beta-talassemia produzem condições clinicamente significativas, como talassemia intermediária de gravidade moderada. Em condição heterozigótica com co-herança de alfa e beta-talassemia, pacientes mostram variabilidade clínica. Assim, nosso objetivo foi a caracterização molecular dos traços da Hb D em individuos clinicamente sintomáticos, devido à co-herança de deleções de alfa ou quaisquer mutações gênicas de beta-globina. TIPO DE ESTUDO E LOCAL: Estudo transversal; realizado em um hospital de cuidado terciário autônomo. MÉTODOS: Hemograma completo e índices de células vermelhas foram medidos pelo analisador automatizado de células. Avaliação quantitativa de hemoglobina Hb F, Hb A, Hb A2 e Hb D foi realizada por cromatografia líquida de alta eficiência. Extração de DNA foi feita pelo método de fenol-clorofórmio. Estudo molecular para deleções comuns de alfa e mutações comuns de beta foi feito por Gap-reação em cadeia da polimerase e amplificação refratária de mutação, respectivamente. RESULTADOS: Avaliamos 30 pacientes e verificamos variação clínica no comportamento dos traços da Hb D. Em seis pacientes, os traços da Hb D foram clinicamente sintomáticos e se comportavam como os de talassemia intermédia. A caracterização molecular mostrou que três desses seis pacientes eram IVS-1-5 positivos. CONCLUSÕES: HPLC pode não ser o padrão ouro para o diagnóstico de traços de Hb D Punjab sintomáticos. Assim, a confirmação padrão ouro deve incluir estudos moleculares.

Genotypic influence of α-deletions on the phenotype of Indian sickle cell anemia patients.

BACKGROUND: Some reports have shown that co-inheritance of α-thalassemia and sickle cell disease improves hematological parameters and results in a relatively mild clinical picture for patients; however, the exact molecular basis and clinical significance of the interaction between α-thalassemia and sickle cell disease in India has not yet been described. There is little agreement on the clinical effects of α-thalassemia on the phenotype of sickle cell disease. METHODS: Complete blood count and red cell indices were measured by an automated cell analyzer. Quantitative assessment of hemoglobin variants HbF, HbA, HbA(2), and HbS was performed by high performance liquid chromatography (HPLC). DNA extraction was performed using the phenol-chloroform method, and molecular study for common α-deletions was done by gap-PCR. RESULTS: Out of 60 sickle cell anemia patients, the α-thalassemia genotype was found in 18 patients. Three patients had the triplicated α-genotype (Anti α-3.7 kb), and the remaining patients did not have α-deletions. This study indicates that patients with co-existing α-thalassemia and sickle cell disease had a mild phenotype, significantly improved hematological parameters, and fewer blood transfusions than the patients with sickle cell anemia without co-existing α-deletions. CONCLUSION: Co-existence of α-thalassemia and sickle cell anemia has significant effects on the phenotype of Indian sickle cell patients.