Immuno-inflammatory predictors of disease severity in COVID-19: A systematic review and meta-analysis.

Deregulated immune response and raised inflammation are the cardinal laboratory features in COVID-19 infection reflecting severity of condition. Detection of the markers will help in early diagnosis with timely therapeutic implementation and effective outcome. Observational studies have suggested alteration in these parameters with severity of the condition. This systematic review and meta-analysis was conducted to assess the relevance of the fact. Observational studies from databases were scrutinised and 3669 articles were identified. Further screening, based on the inclusion criteria a total of 19 articles with 3115 participants, were reviewed for meta-analysis using random effects model. Any data in median and interquartile range were converted to mean ± SD. There was a significant rise in total leukocyte count, C-reactive protein, ferritin, IL-6, IL-10, procalcitonin in severe cases but absolute lymphocyte count, CD4+ and CD8+ registered a fall in severe cases in comparison to non-severe group. Immune and inflammatory markers are significantly altered and related to severity of manifestation in COVID-19 infection.

The dynamics of inflammatory markers in coronavirus disease-2019 (COVID-19) patients: A systematic review and meta-analysis.

BACKGROUND: Coronavirus disease-2019 (COVID-19) is a global pandemic and high mortality rate among severe or critical COVID-19 is linked with SARS-CoV-2 infection-induced hyperinflammation of the innate and adaptive immune systems and the resulting cytokine storm. This paper attempts to conduct a systematic review and meta-analysis of published articles, to evaluate the association of inflammatory parameters with the severity and mortality in COVID-19 patients. METHODS: A comprehensive systematic literature search of medical electronic databases including Pubmed/Medline, Europe PMC, and Google Scholar was performed for relevant data published from January 1, 2020 to June 26, 2020. Observational studies reporting clear extractable data on inflammatory parameters in laboratory-confirmed COVID-19 patients were included. Screening of articles, data extraction and quality assessment were carried out by two authors independently. Standardized mean difference (SMD)/mean difference (MD/WMD) and 95% confidence intervals (CIs) were calculated using random or fixed-effects models. RESULTS: A total of 83 studies were included in the meta-analysis. Of which, 54 studies were grouped by severity, 25 studies were grouped by mortality, and 04 studies were grouped by both severity and mortality. Random effect model results demonstrated that patients with severe COVID-19 group had significantly higher levels of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), procalcitonin (PCT), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-2R (IL-2R), serum amyloid A (SAA) and neutrophil-to-lymphocyte ratio (NLR) compared to those in the non-severe group. Similarly, the fixed-effect model revealed significant higher ferritin level in the severe group when compared with the non-severe group. Furthermore, the random effect model results demonstrated that the non-survivor group had significantly higher levels of CRP, PCT, IL-6, ferritin, and NLR when compared with the survivor group. CONCLUSION: In conclusion, the measurement of these inflammatory parameters could help the physicians to rapidly identify severe COVID-19 patients, hence facilitating the early initiation of effective treatment. PROSPERO REGISTRATION NUMBER: CRD42020193169.

Noncoding RNA Ginir functions as an oncogene by associating with centrosomal proteins.

Long noncoding RNAs constitute a major fraction of the eukaryotic transcriptome, and together with proteins, they intricately fine-tune various growth regulatory signals to control cellular homeostasis. Here, we describe the functional characterisation of a novel pair of long intergenic noncoding RNAs (lincRNAs) comprised of complementary, fully overlapping sense and antisense transcripts Genomic Instability Inducing RNA (Ginir) and antisense RNA of Ginir (Giniras), respectively, from mouse cells. This transcript pair is expressed in a spatiotemporal manner during embryonic development. The individual levels of the sense and antisense transcripts are finely balanced during embryonic growth and in adult tissues. Functional studies of the individual transcripts performed using overexpression and knock-down strategies in mouse cells has led to the discovery that Ginir RNA is a regulator of cellular proliferation and can act as an oncogene having a preeminent role in malignant transformation. Mechanistically, we demonstrate that the oncogenic function of Ginir is mediated by its interaction with centrosomal protein 112 (Cep112). Additionally, we establish here a specific interaction between Cep112 with breast cancer type 1 susceptibility protein (Brca1), another centrosome-associated protein. Next, we prove that the mutual interaction between Cep112 with Brca1 is significant for mitotic regulation and maintenance of genomic stability. Furthermore, we demonstrate that the Cep112 protein interaction with Brca1 protein is impaired when an elevated level of Ginir RNA is present in the cells, resulting in severe deregulation and abnormality in mitosis, leading to malignant transformation. Inhibiting the Ginir RNA function in transformed cells attenuates transformation and restores genomic stability. Together, these findings unravel, to our knowledge, a hitherto-unknown mechanism of oncogenesis mediated by a long noncoding RNA and establishes a unique role of Cep112-Brca1 interaction being modulated by Ginir RNA in maintaining mitotic fidelity.

Study of Red Cell Fragility in Different Stages of Chronic Kidney Disease in Relation to Parathyroid Hormone.

INTRODUCTION: Anaemia is one of the common complications associated with Chronic Kidney Disease (CKD) responsible for the increase in the morbidity and mortality in such patients. Several factors have been attributed to cause renal anaemia, amongst which hyperparathyroidism is one of the less recognised reasons. Most studies have been conducted in this regard in CKD patients undergoing haemodialysis. The level of PTH in early stages of chronic kidney disease has not been much studied. The excess amount of Parathyroid Hormone (PTH) secondary to CKD has been suggested to be a causative factor for anaemia. AIM: To evaluate the serum PTH level in CKD patients before haemodialysis and to study the association of the haemoglobin status with the parathyroid hormone. MATERIALS AND METHODS: Forty CKD patients above 18 years of age before haemodialysis and 25 age and sex matched healthy controls were included in the study. Routine biochemical and haematological parameters such as Routine Blood Sugar (RBS), urea, creatinine, Na+, K+, Ca2+, PTH and Hb% were perfomed. Red cell osmotic fragility was measured by serial dilutions of whole blood with varying concentrations of sodium chloride ranging from 0.1% to 0.9%. RESULTS: The study revealed a significant fall in Hb%, along with a rise in Median Osmotic Fragility (MOF) and PTH in the CKD patients when compared to the control group. Linear regression of PTH with Hb% revealed significant negative association between both the parameters with a R2 value of 0.677. Multilinear regression analysis of MOF and other independent variables such as Hb%, Na+, K+, Ca2+, urea, PTH and creatinine highlighted the variance of MOF by 72%, maximal variance contributed by PTH. Receiver Operating Curve (ROC) analysis revealed an area under the curve of 0.980 with a sensitivity of 100% and specificity of 87% in detecting osmotic fragility at a cut off value of PTH ≥100 pg/ml. CONCLUSION: The underlying cause of anaemia should be identified early in the CKD patients before haemodialysis. Secondary hyperparathyroidism should be ruled out as a causative factor of anaemia to slow down the progression of the disease process.

Extracellular Vesicles As Modulators of Tumor Microenvironment and Disease Progression in Glioma.

Diffuse gliomas are lethal tumors of the central nervous system (CNS) characterized by infiltrative growth, aggressive nature, and therapeutic resistance. The recent 2016 WHO classification for CNS tumors categorizes diffuse glioma into two major types that include IDH wild-type glioblastoma, which is the predominant type and IDH-mutant glioblastoma, which is less common and displays better prognosis. Recent studies suggest presence of a distinct cell population with stem cell features termed as glioma stem cells (GSCs) to be causal in driving tumor growth in glioblastoma. The presence of a stem and progenitor population possibly makes glioblastoma highly heterogeneous. Significantly, tumor growth is driven by interaction of cells residing within the tumor with the surrounding milieu termed as the tumor microenvironment. It comprises of various cell types such as endothelial cells, secreted factors, and the surrounding extracellular matrix, which altogether help perpetuate the proliferation of GSCs. One of the important mediators critical to the cross talk is extracellular vesicles (EVs). These nano-sized vesicles play important roles in intercellular communication by transporting bioactive molecules into the surrounding milieu, thereby altering cellular functions and/or reprogramming recipient cells. With the growing information on the contribution of EVs in modulation of the tumor microenvironment, it is important to determine their role in both supporting as well as promoting tumor growth in glioma. In this review, we provide a comprehensive overview of the role of EVs in tumor progression and glioma pathogenesis.

Evaluation of Lipid Profile and Apolipoproteins in Essential Hypertensive Patients.

INTRODUCTION: Essential hypertension is one of the most common diseases of the Indian population contributing greatly to the morbidity, mortality and economic burden. It has a strong association with cardiovascular disease and abnormal lipid metabolism. Not only the traditional lipid parameters, but also the novel lipid components like Apo A1 and Apo B100 also have been identified to play a role. AIM: The present study was done to evaluate serum lipid profile and Apo A1, Apo B 100 in essential hypertensive patients and correlate their values with the degree of hypertension. MATERIALS AND METHODS: Fasting samples from 55 age and sex matched controls and 55 essential hypertensives were tested for plasma glucose, serum urea, creatinine, lipid profile, apo A1 and apo B100. The cases were subclassified based on the severity of hypertension according to JNC criteria. RESULTS: The study showed a significantly raised value for serum cholesterol, triacylglycerol, Low Density Lipoprotein (LDL), Very Low-Density Lipoprotein (VLDL) in the hypertensive patients than the control group whereas serum High-Density Lipoprotein (HDL) registered a fall in the cases. Apo A1 revealed a non-significant fall in the hypertensive patients. In contrast, there was a rise in the serum apo B100 in the cases. Apo B100/apo A1 ratio was significantly raised in both stage I and stage II hypertensive patients in comparision to the controls. When correlated, serum apo A1 revealed a negative association where as serum apo B 100 showed a positive association with systolic and diastolic bloood pressure. Both LDL/HDL and apoB100/apo A1 and apo B100 revealed a significant positive association with both SBP and DBP. However, apoB100/apo A1 revealed a more positive association in comparision to LDL/HDL ratio (r=0.749, p<0.001, r=0.756, p<0.001 vs r=0.336, p<0.000, r=0.312, p<0.001). CONCLUSION: Apo B100/apoA1 has emerged as an important complementary parameter in addition to traditional lipid ratio for evaluation of risk for future cardiovascular disease.

Apolipoprotein A-I: A Molecule of Diverse Function.

Apolipoprotein A-I (apo A-I) an indispensable component and a major structural protein of high-density lipoprotein (HDL), plays a vital role in reverse cholesterol transport and cellular cholesterol homeostasis since its identification. Its multifunctional role in immunity, inflammation, apoptosis, viral, bacterial infection etc. has crossed its boundary of its potential of protecting cardiovascular system and lowering cardiovascular disease risk, attributing HDL to be known as a protective fat removal particle. Its structural homology with prostacyclin stabilization factor has contributed to its anti-clotting and anti-aggregatory effect on platelet which has potentiated its cardio-protective role as well as its therapeutic efficacy against Alzheimer's disease. The binding affinity and neutralising action against endotoxin lipopolysaccharide, reduces the toxic manifestations of septic shock. As a negative acute phase protein, it blocks T-cell signalling of macrophages. However the recently identified anti-tumor activity of apo A-I has been highlighted in various models of melanoma, lung cancer, ovarian cancer, lymphoblastic leukaemia, gastric as well as pancreatic cancers. These cancer fighting effects are directed towards regression of tumor size and distant metastasis by its immuno modulatory activity as well as its clearing effect on serum lysophospholipids. This lowering effect on lysophospholipid concentration is utilized by apo A-I mimetic peptides to be used in retarding tumor cell proliferation and as a potential cancer therapeutic agent. Not only that, it inhibits the tumor associated neo-angiogenesis as well as brings down the matrix degrading enzymes associated with tumor metastasis. However this efficient therapeutic potential of apo A-I as an anti tumor agent awaits further future experimental studies in humans.

Ischaemic Markers in Acute Hepatic Injury.

INTRODUCTION: Hepatic injury of varied aetiology may progress to Acute Liver Failure (ALF). Compromised microcirculation is thought to be a deciding factor of hepatic hypoxia may be involved in disease progression that needs early detection. Ischaemia markers like serum Ischaemia- modified albumin (IMA), ALT-LDH ratio and ALT-LDH index have been suggested for its detection at early stage. AIM: To find out the association of Ischaemia markers like serum IMA, ALT-LDH ratio and ALT-LDH index in acute hepatic injury cases. MATERIALS AND METHODS: Forty one diagnosed acute liver injury cases of varied aetiology admitted in Department of Medicine, and Gastroenterology of SCB Medical College, Cuttack were enrolled in the study along with 30 age and sex matched healthy controls. Blood collected at time of admission and at time of discharge (1(st) day and 7(th) day) were evaluated for FPG, RFT, LFT, Serum Albumin along with serum LDH, IMA, PT-INR and platelet count. RESULT: Serum bilirubin, hepatic enzymes, IMA, PT-INR was more markedly raised in cases than controls on the 1(st) day of admission. ALT-LDH ratio and index were significantly low in complicated cases. However, on responding to treatment the ALT-LDH index on 7(th) day registered a rise in comparison to the 1(st) day, while serum IMA revealed an insignificant decline showing improvement in hepatic hypoxia. ALT-LDH ratio remains more or less same on response to treatment. CONCLUSION: Serum IMA and ALT-LDH Index reveals association with disease process in Acute Hepatic Injury cases both clinically and biochemically and can be used as supportive parameters for the diagnosis of disease process.

A prototype tobacco-associated oral squamous cell carcinoma classifier using RNA from brush cytology.

BACKGROUND: Oral cancer in the form of squamous cell carcinoma (OSCC) is typically detected in advanced stages when treatment is complex and may not be curative. The need for surgical biopsy may contribute to delays in diagnosis and impede early detection. Multiple studies of RNA from surgically obtained tumor samples have revealed many genes differentially expressed with this disease. We sought to determine whether the identified mRNAs could be used as markers by a non-invasive detection system for OSCC using RNA from brush cytology. METHODS: Levels of mRNAs from 21 genes known to be differentially expressed in head and neck squamous cell carcinoma surgical samples, compared with controls, were shown to be quantifiable in oral brush cytology samples. These mRNAs were quantified in a training set of 14 tumor and 20 non-malignant brush cytology samples from tobacco/betel nut users. With the measurement of two additional mRNAs and analysis using support vector machines algorithm for class prediction of these cancers was produced. RESULTS: This OSCC classifier based on the levels of 5 mRNAs in RNA from brush cytology initially showed 0.93 sensitivity and 0.91 specificity in differentiating OSCC from benign oral mucosal lesions based on leave-one-out cross-validation. When used on a test set of 19 samples from 6 OSCCs and 13 non-malignant oral lesions, we found misclassification of only one OSCC and one benign lesion. CONCLUSIONS: This shows the promise of using RNA from brush cytology for early OSCC detection and the potential for clinical usage of this non-invasive classifier.

Analysis of RNA from brush cytology detects changes in B2M, CYP1B1 and KRT17 levels with OSCC in tobacco users.

RNA expression analysis of oral keratinocytes can be used to detect early oral cancer, but a limitation is the inability to obtain high quality RNA from oral tissue without using biopsies. While oral cytology cell samples can be obtained from patients in a minimally invasive manner, they have not been validated for quantitative analysis of RNA expression. Earlier we showed RNA from brush cytology of hamster Oral Squamous Cell Carcinoma (OSCC) demonstrated differential expression of B2M and CYP1B1 using real time RT-PCR in a dibenz[a,I]pyrene, tobacco carcinogen, induced model of this disease. Here we show reproducibility of this approach to measuring gene expression in humans. Cytology brush samples from 12 tobacco and betel related OSCC and 17 nonmalignant oral lesions revealed B2M mRNA was enriched in tumor samples while CYP1B1 mRNA was reduced, similar to what was seen in the model system. Additionally, we showed that KRT17 mRNA, a gene linked to OSCC in another brush cytology study, was also enriched in OSCC versus nonmalignant lesions, again supporting the promise of using RNA from brush oral cytology to reproducibly monitor oral gene expression.

Brand specific responses to smokeless tobacco in a rat lip canal model.

BACKGROUND: Different compositions of smokeless tobacco (ST) are widely thought to cause oral carcinoma at different rates but there is little direct evidence for this hypothesis. METHODS: We used a rat lip canal model to examine the mucosal changes induced by chronic daily exposure to four different brands of ST: Skoal, Copenhagen, Ettan Swedish Snus, and Stonewall, differing in measured levels of: tobacco specific nitrosamines (TSNAs), unprotonated nicotine, moisture, and pH. RESULTS: Exposure to the lip canal for 12 months produced changes in the mucosa marked by increases in S phase and M phase cells for the Skoal and Copenhagen exposed rats. This correlated with the high level of TSNAs and nicotine in these products. All the tobacco products, to different degrees, induced sites of moderate to severe dysplasia some with extensive rete peg outgrowth from the oral mucosa not seen in the controls. Many of these sites showed a loss of p16 expression. CONCLUSIONS: While all ST products caused dysplasia, the products with lower levels of TSNAs and unprotonated nicotine caused less, consistent with the model that tobacco with low levels of nitrosamines might potentially induce fewer carcinomas in human users.

Effect of Metformin on Hormonal and Biochemical Profile in PCOS Before and After Therapy.

Insulin resistance and the resultant hyperinsulinemia exacerbate the reproductive abnormalities of Polycystic Ovarian Syndrome by increasing ovarian androgen productions and decreasing serum sex hormone binding globulin. The present study was conducted to estimate serum insulin and testosterone level in 44 PCOS cases and 32 control patients. Simultaneously the role of metformin (an insulin sensitizing agent) in modulating insulin resistance and serum androgen level was also analyzed. A significant rise in serum insulin and testosterone (P < 0.001) was observed in cases in comparison to control. Fasting Plasma Glucose to insulin ratio, a marker of insulin resistance revealed a significant fall in PCOS group. Follow up of cases with metformin for 3 months revealed a significant fall in serum insulin (P < 0.05) with improvement in insulin resistance along with a nonsignificant fall in testosterone level. Serum insulin registered a significant positive correlation (P < 0.05) with serum testosterone revealing its etiological association. Thus administration of drugs ameliorating insulin levels is expected to provide new therapeutic modality for PCOS.

Influence of oxidants and anti-oxidants on semen parameters in infertile males.

The deleterious effect of free radicals on spermatozoa was assessed by estimating malondialdehyde and vitamins E and C in seminal plasma and their relation with different sperm parameters. Twenty-two fertile controls with 74 primary infertile males were analysed in the department of biochemistry, SCB Medical College, Cuttack. Seminal malondialdehyde level was observed to be raised in all infertile groups except azoospermic cases in comparison to control. Levels of vitamins E and C were significantly low in the infertile cases (p < 0.001) in relation to control. Significant positive correlation between malondialdehyde with total sperm count and leucocyte count (p < 0.01) indicates their dual contribution towards free radical generation. The negative association of semen malondialdehyde with normal sperm motility and morphology suggests damaging effect of free radicals on sperm membrane integrity. Marked negative correlation of seminal malondialdehyde with both the vitamins signifies their protective utilisation during oxidative phenomenon. In conclusion, understanding the physiologic and pathologic effects of free radicals on sperm function will help in designing new and effective treatment strategies in male infertility.

Modification of the ATM/ATR directed DNA damage response state with aging and long after hepatocyte senescence induction in vivo.

The cellular DNA damage response (DDR) entails the activation of ATM, ATR and/or DNA PK protein kinases that causes modifications of proteins including Chk1, Chk2 and 53BP1, aggregation of DDR proteins into foci, and activation of p53. The DDR is thought to be required for initiation and maintenance of cellular senescence. Potentially senescent cells with DNA damage foci occur in large numbers in vivo with many diseases, but, with the exception of mammalian dermis, there is little evidence for that with normal aging. After experimental induction of cellular senescence in the livers of juvenile mice, there was robust expression of DDR markers in hepatocytes at 1 week; however, by 7 weeks, activation of ATM/ATR kinase targets was limited, although cells with DNA damage foci were present. An analysis of hepatocytes of aged, 22-month-old mice, not experimentally exposed to genotoxins, showed limited activation of ATM/ATR targets, though high numbers of cells with DNA damage foci were found, similar to that seen many weeks after artificial senescence induction in young mice. Based on senescence heterochromatin and SA ss Gal assays of the 22-month-old mouse liver, more than 20% of hepatocytes were potentially senescent, though only some components of the DDR were enriched.

RNA from brush oral cytology to measure squamous cell carcinoma gene expression.

BACKGROUND: RNA expression analysis of oral keratinocytes can be used to detect early stages of disease such as oral cancer or to monitor on-going treatment responses of the same or other oral diseases. A limitation is the inability to obtain high quality RNA from oral tissue without using biopsies. While oral cytology cell samples can be obtained from patients in a minimally invasive manner they have not been validated for quantitative analysis of RNA expression. METHODS: As a starting point in the analysis of tumor markers in oral squamous cell carcinoma (OSCC), we examined RNA in brush cytology samples from hamsters treated with dibenzo[a,l]pyrene to induce oral carcinoma. Three separate samples from each animal were assessed for expression of candidate marker genes and control genes measured with real-time RT-PCR. RESULTS: Brush oral cytology samples from normal mucosa were shown to consist almost exclusively of epithelial cells. Remarkably, ss-2 microglobulin and cytochrome p450, 1B1 (CYP1B1) RNA showed potential utility as markers of OSCC in samples obtained in this rapid and non-surgical manner. CONCLUSION: Brush oral cytology may prove useful as a source of RNA for gene expression analysis during the progression of diseases of the oral epithelium such as OSCC.

Sustained hepatic expression of FoxM1B in transgenic mice has minimal effects on hepatocellular carcinoma development but increases cell proliferation rates in preneoplastic and early neoplastic lesions.

Increased hepatic expression of the Forkhead transcription factor FoxM1B in adult mice accelerates hepatocyte proliferation after partial hepatectomy, while in hepatocytes in intact liver the transgenic (Tg) protein is inactive and has no effect on proliferation. To investigate the influence of FoxM1B on liver tumor formation, we examined the effect of sustained enrichment of FoxM1B in the hepatocytes of mice treated with a diethylnitrosamine (DEN)/phenobarbital tumor induction protocol. Tg enrichment of FoxM1B in hepatocytes did not increase the proliferation rate in normal liver tissue even when the protein was localized to the nucleus. However, it did cause an increase in the proliferation rate and size of preneoplastic and early neoplastic lesions, although having no effects on the total numbers of these lesions. As tumors progressed to hepatocellular carcinomas, the additional Tg FoxM1B protein had no effect on cell proliferation, and there was no increase in tumor burden compared to wild-type animals. This suggests that the artificial enrichment of FoxM1B in the liver, which has been suggested as a gene therapy protocol for liver dysfunction with aging, may not be tumorigenic in that organ.