Immune dysregulation in COVID-19 is the major causal factor associated with disease progression and mortality. Role of monocyte HLA-DR (mHLA-DR), neutrophil CD64 (nCD64) and Immune dysregulation index (IDI) were studied in COVID-19 patients for assessing severity and outcome. Results were compared with other laboratory parameters. Antibody bound per cell for mHLA-DR, nCD64 and IDI were measured in 100 COVID-19 patients by flow cytometry within 12 h of hospital admission. Thirty healthy controls (HC) were included. Clinical and laboratory parameters like C - reactive protein (CRP), Procalcitonin (PCT), Absolute Lymphocyte count (ALC), Absolute Neutrophil count (ANC) and Neutrophil to Lymphocyte ratio (NLR) were recorded. Patients were followed up until recovery with discharge or death. Parameters from 54 mild (MCOV-19), 46 severe (SCOV-19) and 30 HC were analysed. mHLA-DR revealed significant and graded down regulation in MCOV-19 and SCOV-19 as compared to HC whereas IDI was lowest in HC with increasing values in MCOV-19 and SCOV-19. For diagnostic discrimination of MCOV-19 and SCOV-19, IDI revealed highest AUC (0.99). All three immune parameters revealed significant difference between survivors (n = 78) and non-survivors (n = 22). mHLA-DR < 7010 and IDI > 12 had significant association with mortality. Four best performing parameters to identify patients with SCOV-19 at higher risk of mortality were IDI, NLR, ALC and PCT. mHLA-DR and IDI, in addition to NLR and ALC at admission and during hospital stay can be utilized for patient triaging, monitoring, early intervention, and mortality prediction. IDI reported for the first time in this study, appears most promising. Immune monitoring of 'in hospital' cases may provide optimized treatment options.
Supplementary information: The online version contains supplementary material available at 10.1007/s12291-022-01087-z.
{"title":"Monocyte HLADR and Immune Dysregulation Index as Biomarkers for COVID-19 Severity and Mortality.","authors":"Namrata Punit Awasthi, Sridhar Mishra, Vandana Tiwari, Jyotsna Agarwal, Pravin Kumar Das, Paresh Jain, Nuzhat Husain","doi":"10.1007/s12291-022-01087-z","DOIUrl":"https://doi.org/10.1007/s12291-022-01087-z","url":null,"abstract":"<p><p>Immune dysregulation in COVID-19 is the major causal factor associated with disease progression and mortality. Role of monocyte HLA-DR (mHLA-DR), neutrophil CD64 (nCD64) and Immune dysregulation index (IDI) were studied in COVID-19 patients for assessing severity and outcome. Results were compared with other laboratory parameters. Antibody bound per cell for mHLA-DR, nCD64 and IDI were measured in 100 COVID-19 patients by flow cytometry within 12 h of hospital admission. Thirty healthy controls (HC) were included. Clinical and laboratory parameters like C - reactive protein (CRP), Procalcitonin (PCT), Absolute Lymphocyte count (ALC), Absolute Neutrophil count (ANC) and Neutrophil to Lymphocyte ratio (NLR) were recorded. Patients were followed up until recovery with discharge or death. Parameters from 54 mild (MCOV-19), 46 severe (SCOV-19) and 30 HC were analysed. mHLA-DR revealed significant and graded down regulation in MCOV-19 and SCOV-19 as compared to HC whereas IDI was lowest in HC with increasing values in MCOV-19 and SCOV-19. For diagnostic discrimination of MCOV-19 and SCOV-19, IDI revealed highest AUC (0.99). All three immune parameters revealed significant difference between survivors (n = 78) and non-survivors (n = 22). mHLA-DR < 7010 and IDI > 12 had significant association with mortality. Four best performing parameters to identify patients with SCOV-19 at higher risk of mortality were IDI, NLR, ALC and PCT. mHLA-DR and IDI, in addition to NLR and ALC at admission and during hospital stay can be utilized for patient triaging, monitoring, early intervention, and mortality prediction. IDI reported for the first time in this study, appears most promising. Immune monitoring of 'in hospital' cases may provide optimized treatment options.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12291-022-01087-z.</p>","PeriodicalId":13280,"journal":{"name":"Indian Journal of Clinical Biochemistry","volume":"38 2","pages":"204-211"},"PeriodicalIF":2.1,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9540126/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9320026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-01DOI: 10.1007/s12291-022-01059-3
Brajesh Singh, Smiti Singh, J K Bhatia, Rajan Kapoor, Kapil Bhatia
The interrelationship between matrix degradation, oxidative stress, inflammation and trace elements can be speculated in COVID-19. The objective of the study was to evaluate the oxidative stress, inflammation and matrix degradation markers and trace elements in COVID-19 positive patients. A group of confirmed severe COVID-19 positive patients (n = 30) along with COVID-19 negative patients (n = 30) with similar symptoms were included. Both group of patients were assessed for oxidative stress markers, inflammatory cytokines, matrix metalloproteinase (MMP)s and their inhibitors along with trace elements in blood. All the data were subjected to univariate as well as multivariate analysis including PCA, PLS-DA, OPLS-DA. Diagnostic accuracy was tested by ROC curve analysis. Further relationship with Neutrophil/ lymphocyte (N/L) ratio was established if any. Increased oxidative stress, inflammation and matrix degradation is evidenced by significant rise in oxidative markers, inflammatory cytokines and MMP9/TIMP-1 ratio. Decreased Cu/Zn ratio is also observed in COVID-19 positive patients. Multivariate analysis identified SOD, Cu/Zn ratio, IL-6 and TOS, as effective discriminant among the two groups of patients. Further, accuracy was confirmed by ROC curves. Neutrophil/ lymphocyte (N/L) ratio, shows significant negative association with SOD (r= -0.75, p < 0.005) and Cu/Zn ratio (r = -0.88, p < 0.005). These data suggest the attributes of these biomarkers in disease severity. The potential use of these blood-based laboratory markers in disease prognosis seems promising and warrants further attention. Given by the symptoms and severity of the disease, it will be promising to monitor Cu/Zn ratio along with other prognostic indicators.
可以推测COVID-19中基质降解、氧化应激、炎症和微量元素之间的相互关系。本研究的目的是评估COVID-19阳性患者的氧化应激、炎症和基质降解标志物及微量元素。纳入一组确诊的COVID-19严重阳性患者(n = 30)和症状相似的COVID-19阴性患者(n = 30)。对两组患者进行氧化应激标志物、炎症因子、基质金属蛋白酶(MMP)及其抑制剂以及血液中微量元素的检测。所有数据均进行单因素及多因素分析,包括PCA、PLS-DA、OPLS-DA。采用ROC曲线分析检验诊断准确性。进一步确定与中性粒细胞/淋巴细胞(N/L)比值的关系。氧化标记物、炎症因子和MMP9/TIMP-1比值的显著升高证明了氧化应激、炎症和基质降解的增加。在COVID-19阳性患者中也观察到Cu/Zn比降低。多因素分析发现SOD、Cu/Zn比、IL-6和TOS是两组患者的有效判别指标。进一步,通过ROC曲线验证其准确性。中性粒细胞/淋巴细胞(N/L)比值与SOD呈显著负相关(r= -0.75, p
{"title":"Role of Matrix Degradation, Oxidative Stress, Inflammation & Trace Elements in COVID-19 Patients: A Multivariate Study from India.","authors":"Brajesh Singh, Smiti Singh, J K Bhatia, Rajan Kapoor, Kapil Bhatia","doi":"10.1007/s12291-022-01059-3","DOIUrl":"https://doi.org/10.1007/s12291-022-01059-3","url":null,"abstract":"<p><p>The interrelationship between matrix degradation, oxidative stress, inflammation and trace elements can be speculated in COVID-19. The objective of the study was to evaluate the oxidative stress, inflammation and matrix degradation markers and trace elements in COVID-19 positive patients. A group of confirmed severe COVID-19 positive patients (n = 30) along with COVID-19 negative patients (n = 30) with similar symptoms were included. Both group of patients were assessed for oxidative stress markers, inflammatory cytokines, matrix metalloproteinase (MMP)s and their inhibitors along with trace elements in blood. All the data were subjected to univariate as well as multivariate analysis including PCA, PLS-DA, OPLS-DA. Diagnostic accuracy was tested by ROC curve analysis. Further relationship with Neutrophil/ lymphocyte (N/L) ratio was established if any. Increased oxidative stress, inflammation and matrix degradation is evidenced by significant rise in oxidative markers, inflammatory cytokines and MMP9/TIMP-1 ratio. Decreased Cu/Zn ratio is also observed in COVID-19 positive patients. Multivariate analysis identified SOD, Cu/Zn ratio, IL-6 and TOS, as effective discriminant among the two groups of patients. Further, accuracy was confirmed by ROC curves. Neutrophil/ lymphocyte (N/L) ratio, shows significant negative association with SOD (r= -0.75, p < 0.005) and Cu/Zn ratio (r = -0.88, p < 0.005). These data suggest the attributes of these biomarkers in disease severity. The potential use of these blood-based laboratory markers in disease prognosis seems promising and warrants further attention. Given by the symptoms and severity of the disease, it will be promising to monitor Cu/Zn ratio along with other prognostic indicators.</p>","PeriodicalId":13280,"journal":{"name":"Indian Journal of Clinical Biochemistry","volume":"38 2","pages":"193-203"},"PeriodicalIF":2.1,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9446610/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9259278","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
COVID-19, a global pandemic that led to increased morbidity and mortality worldwide since its outcome at the end of the year 2019. A newly discovered variant of severe acute respiratory distress syndrome coronavirus-2 (SARS-CoV-2) was the arbitrator for spreading the syndrome by droplet transmission causing multi-organ failure in many occasions. A post-infection-pro-diabetic disposition was found evident in this study with the persistence of hepato-pancreatic aberrations in respect of reference range of tissue specific bio-markers in hospital admitted COVID-19 cases. The results of this study show that hyperglycemia is a risk factor in precipitating disease oriented complications to the patients with COVID-19 disease. A post-infection follow- up on glycemic-index and related complexities is a vital need to the COVID-19 infected convalescent subjects. Implementation of guidelines on social measure and awareness of anti-viral interventions may be the only way to prevent COVID-19 transmission.
{"title":"A Pilot Study on COVID-19 Positive Subjects: An Excerpt of Post-Infection-Pro-Diabetic Disposition & Related Consequences in Correlation to Hepato-Pancreatic Bio-Markers, Pro-Inflammatory Cytokines and Other Risk Factors.","authors":"Sushil Kumar, Neha Rai, Akash Bansal, Amit Mittal, Nimai Chand Chandra","doi":"10.1007/s12291-022-01054-8","DOIUrl":"https://doi.org/10.1007/s12291-022-01054-8","url":null,"abstract":"<p><p>COVID-19, a global pandemic that led to increased morbidity and mortality worldwide since its outcome at the end of the year 2019. A newly discovered variant of severe acute respiratory distress syndrome coronavirus-2 (SARS-CoV-2) was the arbitrator for spreading the syndrome by droplet transmission causing multi-organ failure in many occasions. A post-infection-pro-diabetic disposition was found evident in this study with the persistence of hepato-pancreatic aberrations in respect of reference range of tissue specific bio-markers in hospital admitted COVID-19 cases. The results of this study show that hyperglycemia is a risk factor in precipitating disease oriented complications to the patients with COVID-19 disease. A post-infection follow- up on glycemic-index and related complexities is a vital need to the COVID-19 infected convalescent subjects. Implementation of guidelines on social measure and awareness of anti-viral interventions may be the only way to prevent COVID-19 transmission.</p>","PeriodicalId":13280,"journal":{"name":"Indian Journal of Clinical Biochemistry","volume":"38 2","pages":"182-192"},"PeriodicalIF":2.1,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9206463/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9266875","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The presence of dyskinesia is the most common side effect of chronic administration of levodopa in Parkinson's disease (PD) subjects. Genetic polymorphisms in levodopa metabolizing gene, catechol-O-methyl transferase (COMT), is shown to influence the inter-individual variability in drug response and adverse events. In the present study, the association of COMT rs6269, rs4633, rs4818, and rs4680 polymorphisms and haplotypes on pharmacokinetics and adverse events with levodopa was investigated in 150 PD patients. The age of onset of PD was 58.00 ± 10 yrs. The most common side effect faced by 78% of the subjects was dyskinesia. The AUC of levodopa was found to be significantly higher in subjects with dyskinesia (1695 ± 113 ng/ml/hr, p < 0.0001) than those without dyskinesia (1550 ± 122 ng/ml/hr). We found that the frequency of subjects presenting dyskinesia was significantly higher in subjects carrying variant genotype of COMT rs6269, rs4633, and rs4680 than that with wild genotype and these subjects presented higher AUC of levodopa. In addition, in subjects with dyskinesia, the AUC of levodopa was found to be significantly higher with low COMT (ACCG) haplotype. The association of COMT rs6269, COMT rs4633, COMT rs4818, and COMT rs4680 variant genotypes with the risk of dyskinesia due to levodopa therapy showed an ROC AUC of 0.67 indicating the moderate prediction of dyskinesia (p = 0.0021) with these COMT variants. In conclusion, PD subjects carrying the variant genotypes of COMT strongly influence high levodopa-induced dyskinesia. Hence the genotyping of COMT before the levodopa therapy will be useful to reduce the adverse events associated with the chronic levodopa treatment.
{"title":"Association of Catechol-O-Methyltransferase Gene Polymorphisms and Haplotypes in the Levodopa-Induced Adverse Events in Subjects with Parkinson's Disease.","authors":"Tasneem Sd Fatima, Syed Tazeem Fathima, Rukmini Mridula Kandadai, Rupam Borgohain, Boddupally Sreenu, Vijay Kumar Kutala","doi":"10.1007/s12291-022-01046-8","DOIUrl":"10.1007/s12291-022-01046-8","url":null,"abstract":"<p><p>The presence of dyskinesia is the most common side effect of chronic administration of levodopa in Parkinson's disease (PD) subjects. Genetic polymorphisms in levodopa metabolizing gene, catechol-O-methyl transferase (COMT), is shown to influence the inter-individual variability in drug response and adverse events. In the present study, the association of COMT rs6269, rs4633, rs4818, and rs4680 polymorphisms and haplotypes on pharmacokinetics and adverse events with levodopa was investigated in 150 PD patients. The age of onset of PD was 58.00 ± 10 yrs. The most common side effect faced by 78% of the subjects was dyskinesia. The AUC of levodopa was found to be significantly higher in subjects with dyskinesia (1695 ± 113 ng/ml/hr, <i>p</i> < 0.0001) than those without dyskinesia (1550 ± 122 ng/ml/hr). We found that the frequency of subjects presenting dyskinesia was significantly higher in subjects carrying variant genotype of COMT rs6269, rs4633, and rs4680 than that with wild genotype and these subjects presented higher AUC of levodopa. In addition, in subjects with dyskinesia, the AUC of levodopa was found to be significantly higher with low COMT (ACCG) haplotype. The association of COMT rs6269, COMT rs4633, COMT rs4818, and COMT rs4680 variant genotypes with the risk of dyskinesia due to levodopa therapy showed an ROC AUC of 0.67 indicating the moderate prediction of dyskinesia (<i>p</i> = 0.0021) with these COMT variants. In conclusion, PD subjects carrying the variant genotypes of COMT strongly influence high levodopa-induced dyskinesia. Hence the genotyping of COMT before the levodopa therapy will be useful to reduce the adverse events associated with the chronic levodopa treatment.</p>","PeriodicalId":13280,"journal":{"name":"Indian Journal of Clinical Biochemistry","volume":"38 2","pages":"262-274"},"PeriodicalIF":2.1,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10070583/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9271403","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<p><p>A substantial group of patients suffer from Covid-19 (CAC) coagulopathy and are presented with thrombosis. The pathogenesis involved in CAC is not fully understood. We evaluated the hemostatic and inflammatory parameters of 51 hospitalized Covid-19 adult patients and 21 controls. The parameters analyzed were danger signal molecule (High molecular weight group box protein-1/HMGBP-1), platelet count, prothrombin time (PT), activated partial thromboplastin time (aPTT), D-dimer, fibrinogen, endothelial protein C receptor (EPCR), soluble E-selectin, soluble P-selectin, thrombomodulin, tissue plasminogen activator (TPA), plasminogen activator inhibitor-1 (PAI-1), soluble fibrin monomer complex (SFMC), platelet-derived microparticles (PDMP), β-thromboglobulin, antithrombin and protein C. The main objective of our study was to investigate which part of the hemostatic system was mostly affected at the admission of Covid-19 patients and whether these parameters could differentiate intensive care unit (ICU) and non-ICU patients. In this prospective case-control study, 51 patients ≥ 18 years who are hospitalized with the diagnosis of Covid-19 and 21 healthy control subjects were included. We divided the patients into two groups according to their medical progress, either in ICU or non-ICU group. Regarding the outcome, patients were again categorized as a survivor and non-survivor groups. Blood samples were collected from patients at admission at the time of hospitalization before the administration of any treatment for Covid-19. The analyzes of the study were made with the IBM SPSS V22 program. <i>p</i> < 0.05 was considered statistically significant. A total of 51 adult patients (F/M: 24/27) (13 ICU and 38 non-ICU) were included in the study cohort. The mean age of the patients was 68.1 ± 14.4 years. The control group consisted of 21 age and sex-matched healthy individuals. All of the patients were hospitalized. In a group of 13 patients, Covid-19 progressed to a severe form, and were hospitalized in ICU. We found out that the levels of fibrinogen, prothrombin time (PT), endothelial protein-C receptor (EPCR), D-dimer, soluble E-selectin, soluble P-selectin, plasminogen activator inhibitor-1 (PAI-1), and tissue plasminogen activator (TPA) were increased; whereas, the levels of soluble fibrin monomer complex (SFMC), platelet-derived microparticles (PDMP), antithrombin and protein-C were decreased in Covid-19 patients compared to the control group at hospital admission. Tissue plasminogen activator was the only marker with a significantly different median level between ICU and non-ICU groups (<i>p</i> < 0.001). In accordance with the previous literature, we showed that Covid-19 associated coagulopathy is distinct from sepsis-induced DIC with prominent early endothelial involvement and fibrinolytic shut-down. Reconstruction of endothelial function at early stages of infection may protect patients from progressing to ICU hospitalization. We believe that af
{"title":"Coagulopathy is Initiated with Endothelial Dysfunction and Disrupted Fibrinolysis in Patients with COVID-19 Disease.","authors":"Fatma Burcu Belen Apak, Gulbahar Yuce, Deniz Ilhan Topcu, Ayse Gultekingil, Yunus Emre Felek, Tugce Sencelikel","doi":"10.1007/s12291-023-01118-3","DOIUrl":"https://doi.org/10.1007/s12291-023-01118-3","url":null,"abstract":"<p><p>A substantial group of patients suffer from Covid-19 (CAC) coagulopathy and are presented with thrombosis. The pathogenesis involved in CAC is not fully understood. We evaluated the hemostatic and inflammatory parameters of 51 hospitalized Covid-19 adult patients and 21 controls. The parameters analyzed were danger signal molecule (High molecular weight group box protein-1/HMGBP-1), platelet count, prothrombin time (PT), activated partial thromboplastin time (aPTT), D-dimer, fibrinogen, endothelial protein C receptor (EPCR), soluble E-selectin, soluble P-selectin, thrombomodulin, tissue plasminogen activator (TPA), plasminogen activator inhibitor-1 (PAI-1), soluble fibrin monomer complex (SFMC), platelet-derived microparticles (PDMP), β-thromboglobulin, antithrombin and protein C. The main objective of our study was to investigate which part of the hemostatic system was mostly affected at the admission of Covid-19 patients and whether these parameters could differentiate intensive care unit (ICU) and non-ICU patients. In this prospective case-control study, 51 patients ≥ 18 years who are hospitalized with the diagnosis of Covid-19 and 21 healthy control subjects were included. We divided the patients into two groups according to their medical progress, either in ICU or non-ICU group. Regarding the outcome, patients were again categorized as a survivor and non-survivor groups. Blood samples were collected from patients at admission at the time of hospitalization before the administration of any treatment for Covid-19. The analyzes of the study were made with the IBM SPSS V22 program. <i>p</i> < 0.05 was considered statistically significant. A total of 51 adult patients (F/M: 24/27) (13 ICU and 38 non-ICU) were included in the study cohort. The mean age of the patients was 68.1 ± 14.4 years. The control group consisted of 21 age and sex-matched healthy individuals. All of the patients were hospitalized. In a group of 13 patients, Covid-19 progressed to a severe form, and were hospitalized in ICU. We found out that the levels of fibrinogen, prothrombin time (PT), endothelial protein-C receptor (EPCR), D-dimer, soluble E-selectin, soluble P-selectin, plasminogen activator inhibitor-1 (PAI-1), and tissue plasminogen activator (TPA) were increased; whereas, the levels of soluble fibrin monomer complex (SFMC), platelet-derived microparticles (PDMP), antithrombin and protein-C were decreased in Covid-19 patients compared to the control group at hospital admission. Tissue plasminogen activator was the only marker with a significantly different median level between ICU and non-ICU groups (<i>p</i> < 0.001). In accordance with the previous literature, we showed that Covid-19 associated coagulopathy is distinct from sepsis-induced DIC with prominent early endothelial involvement and fibrinolytic shut-down. Reconstruction of endothelial function at early stages of infection may protect patients from progressing to ICU hospitalization. We believe that af","PeriodicalId":13280,"journal":{"name":"Indian Journal of Clinical Biochemistry","volume":"38 2","pages":"220-230"},"PeriodicalIF":2.1,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9922102/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9600720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-01-03DOI: 10.1007/s12291-022-01098-w
Sajan Sinha, Atanu Kumar Dutta, Paramita Bhattacharya, Subham Bhattacharya, Mrinal Kanti Das
There is limited data available regarding the clinical utility of routine molecular diagnosis of β Thalassaemia in addition to HPLC-based screening in low resource settings. The current study highlights the caveats of an HPLC-based screening compared to the inclusion of genetic confirmation as a second-tier test and its implications in terms of genotype-phenotype correlation. A prospective, institution-based, observational study was conducted at the Department of Paediatric Medicine, including 103 children aged up to 12 years. Five common mutations for β Thalassemia and the HbE mutation in the HBB gene were tested by a two-tiered approach using multiplex ARMS PCR and PCR RFLP methods respectively. Sanger sequencing of all three exons of the HBB gene was performed in all negative cases. Sequencing revealed many rare pathogenic mutations like c.316-106 C > G (dbSNP: 34,690,599); Hb Kairouan (c.92G > C); c.33 C > A (dbSNP rs35799536); c.47G > A (dbSNP rs63750783); c.51delC (HbVar ID 799); c.[93-2 A > C] and c.118 C > T (HbVar ID 845). We detected a novel Pathogenic M_000518.5(HBB):c.164_168delinsGGCATCA (p.Val55fs) mutation in a heterozygous state which was reported in the ClinVar database with accession ID VCV000590977.2. We also encountered several cases of silent carrier on HPLC and de novo occurrence of mutation. We conclude that the multiplex touchdown ARMS PCR methodology employed in the present study provides a low-cost solution for molecular diagnostics of Β Thalassaemia. The problem of silent carriers in HPLC is significant enough to rethink if we need supplemental genetic testing in the couple when one of the partners is a carrier.
在资源匮乏的地区,除了基于高效液相色谱的筛查外,关于β地中海贫血常规分子诊断的临床应用数据有限。目前的研究强调了以高效液相色谱为基础的筛查与将遗传确认作为第二级检测的注意事项及其在基因型-表型相关性方面的意义。在儿科医学系进行了一项前瞻性、基于机构的观察性研究,包括103名12岁以下的儿童。采用多重ARMS PCR和PCR RFLP两种方法分别检测β地中海贫血的5种常见突变和HBB基因中的HbE突变。在所有阴性病例中,对HBB基因的所有三个外显子进行Sanger测序。测序显示许多罕见的致病突变,如C .316-106 C > G (dbSNP: 34,690,599);Hb Kairouan (C . 92g > C);c.33C > A (dbSNP rs35799536);c.47G > A (dbSNP rs63750783);c.51delC (HbVar ID 799);c.[93-2 A > c]C > T (HbVar ID 845)。我们检测到一种新的致病因子M_000518.5(HBB):c。在ClinVar数据库中报道了一个杂合状态的164_168delinsGGCATCA (p.Val55fs)突变,登录ID为VCV000590977.2。我们还在HPLC上遇到了几例沉默携带者和重新发生突变的病例。我们得出结论,本研究中采用的多重触点ARMS PCR方法为Β地中海贫血的分子诊断提供了一种低成本的解决方案。HPLC中沉默携带者的问题非常重要,足以让我们重新思考,当其中一方是携带者时,我们是否需要对夫妇进行补充基因检测。
{"title":"Spectrum of Rare and Novel Indel Mutations Responsible for β Thalassemia in Eastern India","authors":"Sajan Sinha, Atanu Kumar Dutta, Paramita Bhattacharya, Subham Bhattacharya, Mrinal Kanti Das","doi":"10.1007/s12291-022-01098-w","DOIUrl":"https://doi.org/10.1007/s12291-022-01098-w","url":null,"abstract":"There is limited data available regarding the clinical utility of routine molecular diagnosis of β Thalassaemia in addition to HPLC-based screening in low resource settings. The current study highlights the caveats of an HPLC-based screening compared to the inclusion of genetic confirmation as a second-tier test and its implications in terms of genotype-phenotype correlation. A prospective, institution-based, observational study was conducted at the Department of Paediatric Medicine, including 103 children aged up to 12 years. Five common mutations for β Thalassemia and the HbE mutation in the HBB gene were tested by a two-tiered approach using multiplex ARMS PCR and PCR RFLP methods respectively. Sanger sequencing of all three exons of the HBB gene was performed in all negative cases. Sequencing revealed many rare pathogenic mutations like c.316-106 C > G (dbSNP: 34,690,599); Hb Kairouan (c.92G > C); c.33 C > A (dbSNP rs35799536); c.47G > A (dbSNP rs63750783); c.51delC (HbVar ID 799); c.[93-2 A > C] and c.118 C > T (HbVar ID 845). We detected a novel Pathogenic M_000518.5(HBB):c.164_168delinsGGCATCA (p.Val55fs) mutation in a heterozygous state which was reported in the ClinVar database with accession ID VCV000590977.2. We also encountered several cases of silent carrier on HPLC and de novo occurrence of mutation. We conclude that the multiplex touchdown ARMS PCR methodology employed in the present study provides a low-cost solution for molecular diagnostics of Β Thalassaemia. The problem of silent carriers in HPLC is significant enough to rethink if we need supplemental genetic testing in the couple when one of the partners is a carrier.","PeriodicalId":13280,"journal":{"name":"Indian Journal of Clinical Biochemistry","volume":"8 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135554720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cell-free DNA (cfDNA) is released into the plasma of patients with cardiac disease. Here, the source and mechanism of plasma cfDNA release in patients with myocardial infarction (MI) and other cardiac diseases (n = 59) were investigated. Plasma levels of various markers including M30 (apoptosis), M65 (apoptosis and necrosis), cyclophilin A (CyPA) (necrosis), and myeloperoxidase (MPO) (neutrophil activation) were assayed. The plasma cfDNA concentrations in MI and other cardiac diseases were significantly higher than that in the healthy control subjects. Significant differences were not observed among the cardiac disease patients (MI and other cardiac diseases) and healthy control subjects in M30, M65, and CyPA levels. In contrast,the MPO levels were significantly elevated in cardiac disease patients when compared to control groups, and MPO levels in MI patients were significantly higher than other cardiac diseases patients. These results suggest that cfDNA is mainly released by neutrophils via NETosis in addition to apoptosis except for epithelial apoptosis in patients with cardiac disease and the degree is greater in MI patients. The results from this study provide basic information for diagnosis marker of MI.
{"title":"Cell-free DNA Release in the Plasma of Patients with Cardiac Disease is Associated with Cell Death Processes.","authors":"Junko Fujihara, Yoshikazu Takinami, Kaori Kimura-Kataoka, Yasuyuki Kawai, Haruo Takeshita","doi":"10.1007/s12291-022-01034-y","DOIUrl":"10.1007/s12291-022-01034-y","url":null,"abstract":"<p><p>Cell-free DNA (cfDNA) is released into the plasma of patients with cardiac disease. Here, the source and mechanism of plasma cfDNA release in patients with myocardial infarction (MI) and other cardiac diseases (<i>n</i> = 59) were investigated. Plasma levels of various markers including M30 (apoptosis), M65 (apoptosis and necrosis), cyclophilin A (CyPA) (necrosis), and myeloperoxidase (MPO) (neutrophil activation) were assayed. The plasma cfDNA concentrations in MI and other cardiac diseases were significantly higher than that in the healthy control subjects. Significant differences were not observed among the cardiac disease patients (MI and other cardiac diseases) and healthy control subjects in M30, M65, and CyPA levels. In contrast,the MPO levels were significantly elevated in cardiac disease patients when compared to control groups, and MPO levels in MI patients were significantly higher than other cardiac diseases patients. These results suggest that cfDNA is mainly released by neutrophils via NETosis in addition to apoptosis except for epithelial apoptosis in patients with cardiac disease and the degree is greater in MI patients. The results from this study provide basic information for diagnosis marker of MI.</p>","PeriodicalId":13280,"journal":{"name":"Indian Journal of Clinical Biochemistry","volume":"38 1","pages":"67-72"},"PeriodicalIF":1.5,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9852365/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10578525","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abutilon indicum (L) is an Indian traditional plant used for the treatment of diabetes and heart diseases. The present study is to evaluate the functional of A. indicum leaf extract as insulin like character to inhibit lipolysis and stimulates Adipogenesis activity. The ability of the A. indicum leaf extract in anti-obesity effect of Adipogenesis, lipolysis and cholesterol esterase functions can be predicted by using 3T3-L1 adipocyte cell lines. Substances were isolated from A. indicum leaves and the double filtered crude sample were used for Adipogenesis, lipolysis and cholesterol esterase activity using 3T3-L1 adipocytes at different concentrations. We used differential media-I, differential media-II and maintenance media (MM1) at concentrations of 20, 40, 60, 80, 100, 200 and 400 µg/mL respectively. In addition to the extract, there is a significance increase in glycerol release (p < 0.001) compared with crude and reference compounds. Cholesterol esterase activity predicts the IC50 = 27.11 µg/mL of orlistat positive control compare with IC50 = 8.158 µg/mL of crude extract. Based on the observation, A. indicum leaf extract can promotes lipolysis and differentiated adipocytes. It is potentially used as adjuvant in the treatment of Type 2 diabetes.
{"title":"Effect of <i>Abutilon indicum</i> (L) Extract on Adipogenesis, Lipolysis and Cholesterol Esterase in 3T3-L1 Adipocyte Cell Lines.","authors":"Lavanya Lakshminarayana, V Veeraraghavan, Kuruvalli Gouthami, Renuka Srihari, Prashantha Chowdadenahalli Nagaraja","doi":"10.1007/s12291-022-01022-2","DOIUrl":"10.1007/s12291-022-01022-2","url":null,"abstract":"<p><p><i>Abutilon indicum</i> (L) is an Indian traditional plant used for the treatment of diabetes and heart diseases. The present study is to evaluate the functional of <i>A. indicum</i> leaf extract as insulin like character to inhibit lipolysis and stimulates Adipogenesis activity. The ability of the <i>A. indicum</i> leaf extract in anti-obesity effect of Adipogenesis, lipolysis and cholesterol esterase functions can be predicted by using 3T3-L1 adipocyte cell lines. Substances were isolated from <i>A. indicum</i> leaves and the double filtered crude sample were used for Adipogenesis, lipolysis and cholesterol esterase activity using 3T3-L1 adipocytes at different concentrations. We used differential media-I, differential media-II and maintenance media (MM1) at concentrations of 20, 40, 60, 80, 100, 200 and 400 µg/mL respectively. In addition to the extract, there is a significance increase in glycerol release (<i>p</i> < 0.001) compared with crude and reference compounds. Cholesterol esterase activity predicts the IC<sub>50</sub> = 27.11 µg/mL of orlistat positive control compare with IC<sub>50</sub> = 8.158 µg/mL of crude extract. Based on the observation, <i>A. indicum</i> leaf extract can promotes lipolysis and differentiated adipocytes. It is potentially used as adjuvant in the treatment of Type 2 diabetes.</p>","PeriodicalId":13280,"journal":{"name":"Indian Journal of Clinical Biochemistry","volume":"38 1","pages":"22-32"},"PeriodicalIF":2.1,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9852410/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10580743","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-01-01Epub Date: 2021-03-25DOI: 10.1007/s12291-021-00970-5
Mala Mahto, Ayan Banerjee, Mukunda Kumar, Sushil Kumar, Jagjit Pandey
Varying reports across different laboratories or across different analysers in the same lab for the same sample is not an uncommon phenomena. Experts call this a lack of harmonization. A test that is harmonized provides the same results regardless of the manufacturer of reagents used or the laboratory where the test is performed. When laboratory tests are not harmonized, the entire continuum of patient care can be affected in a number of ways. Here, we present a case of varying reports for a single serum human chorionic gonadotropin (hCG) sample on two different immunoassay platforms for a young female presenting with an abdominopelvic mass. The lab reports for serum hCG for this particular patient showed inconsistent results with the same sample within the same lab. The phenomena behind this was lack of harmonization of test results. We introspect many of the factors responsible for lack of uniformity in hCG results amongst the major ones being with use of antibodies directed against different epitopes of hCG (analyte) and the heterogeneity of the hCG molecule itself. Harmonization is a process to ensure that different clinical testing procedures used by different laboratories give equivalent results. Harmonizing test results will enable healthcare providers to use clinical guidelines with greater confidence for diagnosing disease and managing patients.
{"title":"A Search for Uniformity in Human Chorionic Gonadotropin (hCG) Reporting.","authors":"Mala Mahto, Ayan Banerjee, Mukunda Kumar, Sushil Kumar, Jagjit Pandey","doi":"10.1007/s12291-021-00970-5","DOIUrl":"10.1007/s12291-021-00970-5","url":null,"abstract":"<p><p>Varying reports across different laboratories or across different analysers in the same lab for the same sample is not an uncommon phenomena. Experts call this a lack of harmonization. A test that is harmonized provides the same results regardless of the manufacturer of reagents used or the laboratory where the test is performed. When laboratory tests are not harmonized, the entire continuum of patient care can be affected in a number of ways. Here, we present a case of varying reports for a single serum human chorionic gonadotropin (hCG) sample on two different immunoassay platforms for a young female presenting with an abdominopelvic mass. The lab reports for serum hCG for this particular patient showed inconsistent results with the same sample within the same lab. The phenomena behind this was lack of harmonization of test results. We introspect many of the factors responsible for lack of uniformity in hCG results amongst the major ones being with use of antibodies directed against different epitopes of hCG (analyte) and the heterogeneity of the hCG molecule itself. Harmonization is a process to ensure that different clinical testing procedures used by different laboratories give equivalent results. Harmonizing test results will enable healthcare providers to use clinical guidelines with greater confidence for diagnosing disease and managing patients.</p>","PeriodicalId":13280,"journal":{"name":"Indian Journal of Clinical Biochemistry","volume":"38 1","pages":"142-147"},"PeriodicalIF":2.1,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s12291-021-00970-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10580746","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-01-01Epub Date: 2021-02-11DOI: 10.1007/s12291-021-00957-2
Sudhesna Mohapatra, Sutirtha Chakraborty
Thyroid stimulating hormone (TSH) immunoassays are known for giving varying results based on the platform of testing and the generation of kit used. It is generally expected that the results should not vary to affect clinical diagnosis and management. We aimed to perform method comparison study between two TSH assays by the same manufacturer Siemens Healthineers. Results show that there is a large proportional error between the assays with a bias of -3.71mIu/L indicating that TSH assay gives higher values for TSH for the same patient as measured against the TSH3-Ultra kit. This can affect interpretation of results leading to false increase in patients categorized under hypothyroidism and subclinical hypothyroidism. We strongly suggest, to prevent errors in clinical evaluation of a patient with thyroid dysfunction, validation of the performance of the assay and method comparison should be performed in-house.
{"title":"Analytical Variation Between Two Different TSH Reagents from the Same Manufacturer.","authors":"Sudhesna Mohapatra, Sutirtha Chakraborty","doi":"10.1007/s12291-021-00957-2","DOIUrl":"10.1007/s12291-021-00957-2","url":null,"abstract":"<p><p>Thyroid stimulating hormone (TSH) immunoassays are known for giving varying results based on the platform of testing and the generation of kit used. It is generally expected that the results should not vary to affect clinical diagnosis and management. We aimed to perform method comparison study between two TSH assays by the same manufacturer Siemens Healthineers. Results show that there is a large proportional error between the assays with a bias of -3.71mIu/L indicating that TSH assay gives higher values for TSH for the same patient as measured against the TSH3-Ultra kit. This can affect interpretation of results leading to false increase in patients categorized under hypothyroidism and subclinical hypothyroidism. We strongly suggest, to prevent errors in clinical evaluation of a patient with thyroid dysfunction, validation of the performance of the assay and method comparison should be performed in-house.</p>","PeriodicalId":13280,"journal":{"name":"Indian Journal of Clinical Biochemistry","volume":"38 1","pages":"132-135"},"PeriodicalIF":2.1,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1007/s12291-021-00957-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10585899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}