Identification of Clinical Response Predictors of Tocilizumab Treatment in Patients with Severe COVID-19 Based on Single-Center Experience.

Hyperinflammation in COVID-19 plays a crucial role in pathogenesis and severity; thus, many immunomodulatory agents are applied in its treatment. We aimed to identify good clinical response predictors of tocilizumab (TCZ) treatment in severe COVID-19, among clinical, laboratory, and radiological variables. We conducted a prospective, observational study with 120 patients with severe COVID-19 not improving despite dexamethasone (DEX) treatment. We used parametric and non-parametric statistics, univariate logistic regression, receiver operating characteristic (ROC) curves, and nonlinear factors tertile analysis. In total, 86 (71.7%) patients achieved the primary outcome of a good clinical response to TCZ. We identified forty-nine predictive factors with potential utility in patient selection and treatment monitoring. The strongest included time from symptom onset between 9 and 12 days, less than 70% of estimated radiological lung involvement, and lower activity of lactate dehydrogenase. Additional predictors were associated with respiratory function, vitamin D concentration, comorbidities, and inflammatory/organ damage biomarkers. Adverse events analysis proved the safety of such a regimen. Our study confirmed that using TCZ early in the hyperinflammatory phase, before severe respiratory failure development, is most beneficial. Considering the described predictive factors, employing simple and widely available laboratory, radiological, and clinical tools can optimize patient selection for immunomodulatory treatment with TCZ.

On-admission laboratory predictors for developing critical COVID-19 during hospitalization - a multivariable logistic regression model.

INTRODUCTION AND OBJECTIVE: Recognition of patients with COVID-19 who will progress clinically and need respiratory support remains challenging. The aim of the study was to identify abnormalities in on-admission laboratory results that can precede progression from moderate or severe to critical COVID-19. MATERIAL AND METHODS: Laboratory data analyzed of 190 patients admitted with moderate or severe COVID-19 to our ward. Laboratory results taken into analysis were obtained during the first 48 hours of hospitalization. Multivariate logistic regression was performed using risk factors obtained in the univariate analysis as dependent variables. RESULTS: 42 patients were identified who developed critical COVID-19. In univariate analysis, 22 laboratory risk factors were detected that were used in logistic regression and in building model with following predictors: high-sensitive troponin I concentration (hs-TnI) >26 ng/mL (OR 13.45; 95%CI 3.28-55.11; P 15 (OR 5.67; 95%CI 1.97-16.36, P 50 pg/mL (OR 5.52; 95%CI 1.86-16.37; P = 0.001), fasting glycaemia >6.8 mmol/L (OR 4.74; 95%CI 1.65-13.66; P = 0.002), immature neutrophils count >0.06/µL (OR 4.06; 95%CI 1.35-12.2; P = 0.012) and urine protein concentration >500 mg/L (OR 2.94; 95%CI 1.04-8.31; P = 0.043). CONCLUSIONS: The most significant risk factors of developing critical COVID-19 during hospitalization are: elevated hs-TnI, IL-6, and glucose serum concentrations, increased immature neutrophil count, neutrophils to monocytes ratio, and proteinuria during the first 48 hours after admission. The model built with these predictors achieved better predictive performance than any other univariately analysed laboratory markers in predicting the critical development COVID-19.

Predominance of genotype P[9]G3 in rotavirus gastroenteritis in Polish children.

INTRODUCTION: Rotavirus (RV) infection is the most common cause of gastroenteritis in children. This paper identifies the most common genotypes of rotaviruses isolated from children hospitalized with gastroenteritis and attempts to determine any relationship between infection with a certain rotavirus genotype. MATERIAL AND METHODS: The investigated group consisted of 68 consecutive children with rotavirus gastroenteritis (confirmed by an agglutination test). Rotavirus genotype was determined in stool samples obtained from each child. RESULTS: The P[9]VP4 genotype was observed in 41/61 positive samples (over 67.2%) that were permanently associated with the G3 VP7 genotype. Moreover, G3 was determined as the most commonly isolated G type (77.94%). As well as the P[9]G3 type, G3 was also found in the P[4] type (5 cases). Twenty-six out of 61 (42.6%) children in whom rotavirus genotype was determined were co-infected with pathogenic bacteria. No statistical correlation was observed between rotavirus P[9]G3 gastroenteritis and digestive tract co-infection with pathogenic bacteria (p > 0.05). Elevated ALT activity was found in 34/59 (57.6%) cases of rotavirus gastroenteritis. Elevated ALT serum level was found to correlate with P[9]G3 rotavirus genotype but concomitant infections did not. CONCLUSIONS: The most common genotype of rotaviruses observed in our group of children, P[9]G3, has rarely been described. Co-infection of the digestive tract with pathogenic bacteria and elevated serum ALT concentrations were found to be the most frequent phenomena. A correlation between P[9]G3 rotavirus genotype and elevated serum ALT level was found, but no significant relationship was identified between concomitant infections and P[9]G3 genotype.

Decreased level of intracellular cholesterol in peripheral blood mononuclear cells is associated with chronic hepatitis C virus infection.

Peripheral blood mononuclear cells (PBMCs) constitute the main extrahepatic place of, hepatitis C virus (HCV) replication. We aimed to determine the impact of CHC infection and microRNA-, 122 expression on cholesterol expression in PBMCs. HCV RNA strand, intracellular cholesterol, HMGCoA, reductase and miR-122 expression in PBMC were determined in 54 CHC patients. The study shows that significant decrease of intracellular cholesterol level in PBMC (p=0.000000), accompanied by serum hypocholesterolemia is the characteristic feature of chronic hepatitis C infection. Although, microRNA-122 expression was detectable in PBMCs of CHC patients (52.5%), the alteration of intracellular cholesterol level was independent of miR-122 expression.

[Influence of the selected pyrimidine compounds on the activity of thymidine phosphorylase from normal and tumor endometrial cells]. / Wplyw wybranych zwiazków pirymidynowych na aktywnosc fosforylazy tymidynowej komórek endometrialnych prawidlowych i nowotworowych.

OBJECTIVES: The aim of this study was to evaluate the influence of the selected pyrimidine compounds on the activity of thymidine phosphorylase (TP) of normal and tumor endometrial cells. MATERIALS AND METHODS: Influence of 28 chemical compounds on the TP activity in the cytosol of the endometrial cells was studied by the spectrophotometric method. The studied group comprised postmenopausal women with endometrial cancer: adenocarcinoma endometrialis (Adeno Ca E). The second group included women with normal endometrium after surgery due to non-oncologic reasons. RESULTS: The most potent inhibitor of TP activity from cancer and endometrium was synthesized 5-bromo-6-acetyloaminouracil, which at the 0.2 mM concentration, by 0.2 mM concentration thymidine reduced the cytosol TP activity by about 80%. 5-bromo-6-aminouracil, 5-nitrouracil and 5-bromouracil reduced this TP activity in statistically significant manner. From among synthesized 1-N-allyloxymethylpyrimidine derivatives 1-N-allyloxymethylthymine was the strongest inhibitor of the TP activity in endometrium, and 1-N-allyloxymethyl-4-hydrokxy-5-nitro-6-oxopyrimidine in endometrial cancer respectively. The most potent activators of TP in endometrial cancer was 5-bromodeoxyuridine and 1-N-allyloxymethyl-5-nitrouracil, which increased the TP activity about 100%. 5-fluorodeoxyuridine, 5-jododeoxyuridine and 2'-deoxyuridine activated the TP in statistically significant manner too, but stronger in case of endometrial cancer than in normal endometrium. The synthesized 5-bromo-6-acetyloaminouracil strongly inhibited the TP activity of endometrial cells and might be useful in reducing endometrial cancer angiogenesis. On the other hand 5-bromodeoxyuridine and the synthesized 1-N-allyloxymethyl-5-nitrouracil might increase the effect of antitumor therapy with the cytostatics. These conclusions ought to be confirmed by analyzing more tumor cases.

Mevalonate pathway modulation is associated with hepatitis C virus RNA presence in peripheral blood mononuclear cells.

Peripheral blood mononuclear cells (PBMC) constitute the main extrahepatic reservoir of hepatitis C virus (HCV). Lipid metabolism of host seems to play important role in HCV infection. The relationship between HCV presence in PBMC and the expression of mevalonate pathway has not been elucidated. The aim of this study was to investigate the association between mevalonate pathway and HCV RNA presence in PBMC after anti-HCV treatment. 67 serum and corresponding PBMC samples were collected from patients at the end of interferon alpha and ribavirin treatment. Serum total cholesterol, HDL-C and LDL-C fractions, triglycerides, as well as intracellular cholesterol and expression level of HMG-CoA reductase, geranylgeranyl pyrophosphate synthase in PBMC were measured and matched for the HCV RNA presence or absence in sera/PBMC. HCV RNA elimination from sera and PBMC was associated with higher serum cholesterol (118.5mg/dL) and LDL-C (66.42mg/dL) levels, compared to the group, where HCV RNA was detected only in PBMC (100.94 and 53.22mg/dL) or the group, where HCV RNA was found in both sera and PBMC (86.79 and 43.79mg/dL) after treatment. Increased expression of geranylgeranyl pyrophosphate synthase was found in the majority of PBMC samples that harbored HCV RNA after elimination of HCV RNA from sera. The expression of mevalonate pathway after antiviral treatment seems to be modulated depending on HCV RNA status in peripheral blood mononuclear cells.

[Late spontaneous elimination of HCV-RNA from mononuclear cells derived from peripheral blood in patients with chronic hepatitis C]. / Pózna samoistna eliminacja HCV-RNA z komórek jednojadrzastych krwi obwodowej po skutecznym leczeniu przewleklego zapalenia watroby typu C.

THE AIM: To follow persistence of HCV-RNA in PBMC in patients with chronic hepatitis C (CHC). To estimate the influence of this phenomenon on the cellular immune response of peripheral blood lymphocytes. MATERIAL AND METHODS: 8 HCV-RNA in PBMC positive children, with undetectable serum HCV-RNA after antiviral treatment, have been examined every 2-3 years. The amount of IFN-gamma, IL-12 and IL-18 secreted by PBMC obtained from the children after stimulation with phytohemagglutinin (PHA) was measured. RESULTS: Spontaneous elimination of HCV-RNA from PBMC in 2 to 6 years after treatment was found in all children. In two children HCV-RNA detectable both in serum and in PBMC, without recurrence of hepatitis, was found in single examination. PBMC containing HCV-RNA secreted more IFN-gamma than PBMC lacking it (1221 +/- 458 pg/ml vs. 651 +/- 147 pg/ml; p=0.009, similar correlation was revealed with the regard of IL-12: 21.8 +/- 12.3 pg/ml vs. 5,6 +/- 3,3 pg/ml respectively; p=0.009. Production and release of IL-18 were not correlated with HCV-RNA persistence (p=0.12). CONCLUSION: Patients with CHC and persistence of HCV-RNA in PBMC require longitudinal follow-up in the respect of possible reseroconversion. PBMC containing HCV-RNA reveal enhanced cellular immune response, which most probably effects in spontaneous elimination of the virus.

A competitor DNA template for the molecular quantification of the hepatitis B virus.

The molecular determination of viral load in the serum represents the most valuable prognostic marker of HBV infection. In this paper, a new molecular assay for the quantitative measurement of HBV presence is described. It is based on PCR performing with a HBV-specific competitor DNA template. For the construction of the DNA template, a HBV DNA-originated 436 bp DNA fragment was modified by introducing a 110 bp deletion and cloned into pUC19. The resulting vector serves as the competitor DNA template in the competitive PCR. Post-PCR, the competitor DNA generates an amplified fragment of 306 bp; it could be easily distinguished from the product generated from the viral-originated DNA product (416 bp) when the same primers are used. The quantitative ratio between the two products enables the quantitative determination of viral load. The range of the HB-PCR assay is from 3 x 10(4)to 6 x 10(10) particles/ml. A serum HBV load determination performed by HB-PCR assay indicated a close correlation with the results of the Quantiplex HBV DNA assay (bDNA). The HB-PCR assay is cheap, reliable and easy to use in any laboratory working with PCR methods.