Multicenter analysis of neutrophil extracellular trap dysregulation in adult and pediatric COVID-19

Dysregulation in neutrophil extracellular trap (NET) formation and degradation may play a role in the pathogenesis and severity of COVID-19; however, its role in the pediatric manifestations of this disease including MIS-C and chilblain-like lesions (CLL), otherwise known as "COVID toes", remains unclear. Studying multinational cohorts, we found that, in CLL, NETs were significantly increased in serum and skin. There was geographic variability in the prevalence of increased NETs in MIS-C, in association with disease severity. MIS-C and CLL serum samples displayed decreased NET degradation ability, in association with C1q and G-actin or anti-NET antibodies, respectively, but not with genetic variants of DNases. In adult COVID-19, persistent elevations in NETs post-disease diagnosis were detected but did not occur in asymptomatic infection. COVID-19-affected adults displayed significant prevalence of impaired NET degradation, in association with anti-DNase1L3, G-actin, and specific disease manifestations, but not with genetic variants of DNases. NETs were detected in many organs of adult patients who died from COVID-19 complications. Infection with the Omicron variant was associated with decreased levels of NETs when compared to other SARS-CoV-2 strains. These data support a role for NETs in the pathogenesis and severity of COVID-19 in pediatric and adult patients. SummaryNET formation and degradation are dysregulated in pediatric and symptomatic adult patients with various complications of COVID-19, in association with disease severity. NET degradation impairments are multifactorial and associated with natural inhibitors of DNase 1, G-actin and anti-DNase1L3 and anti-NET antibodies. Infection with the Omicron variant is associated with decreased levels of NETs when compared to other SARS-CoV-2 strains.

A Working Model to Inform Risk-Based Back to Work Strategies

BackgroundThe coronavirus disease 2019 (COVID-19) pandemic has forced many businesses to close or move to remote work to reduce the potential spread of disease. Employers desiring a return to onsite work want to understand their risk for having an infected employee on site and how best to mitigate this risk. Here, we modelled a range of key metrics to help inform return to work policies and procedures, including evaluating the benefit and optimal design of a SARS-CoV-2 employee screening program. MethodsWe modeled a range of input variables including prevalence of COVID-19, time infected, number of employees, test sensitivity and specificity, test turnaround time, number of times tested within the infectious period, and sample pooling. We modeled the impact of these input variables on several output variables: number of healthy employees; number of infected employees; number of test positive and test negative employees; number of true positive, false positive, true negative, and false negative employees; positive and negative predictive values; and time an infected, potentially contagious employee is on site. ResultsWe show that an employee screening program can reduce the risk for onsite transmission across different prevalence values and group sizes. For example, at a pre-test asymptomatic community prevalence of 0.5% (5 in 1000) with an employee group size of 500, the risk for at least one infected employee on site is 91.8%, with 3 asymptomatic infected employees predicted within those 500 employees. Implementing a SARS-CoV-2 baseline screen with an 80% sensitivity and 99.5% specificity would reduce the risk of at least one infected employee on site to 39.4% and the predicted number of infected employees onsite (false negatives) to 1. Repetitive testing is required for ongoing vigilance of onsite employees. The expected number of days an infected employee is on site depends on test sensitivity, testing interval, and turnaround time. If the test interval is longer than the infectious period ([~]14 days for COVID-19), testing will not detect the infected employee. Sample pooling reduces the number of tests performed, thereby reducing testing costs. However, the pooling methodology (eg, 1-stage vs 2-stage pooling, pool size) will impact the number of employees that screen positive, thereby affected the number of employees eligible to return to onsite work. ConclusionsThe modeling presented here can be used to help employers understand their risk for having an infected employee on site. Further, it details how an employee screening program can reduce this risk and shows how screening performance and frequency impact the effectiveness of a screening program. The primary factors determining the effectiveness of a screening program are test sensitivity and frequency of testing. DisclaimerThis publication is offered to businesses/employers as a model of potential risk arising from COVID19 in the workplace. While believed to be based on reliable data, the model described herein has not been prospectively validated and should not be relied upon for any purpose other than as an aid to understand the potential impacts of a number of variables on the risk of having COVID19 positive employees on a worksite. Decisions related to workplace safety; COVID19 related workplace testing; programs and procedures should be based upon your actual data and applicable laws and public health orders.

Clinicopathological and prognostic significance of serum and tissue Dickkopf-1 levels in human hepatocellular carcinoma.

BACKGROUND: Although Dickkopf-1 (DKK1) is known to be a negative regulator of the Wnt/ß-catenin pathway, it has been recently found to be upregulated in cancers. AIMS: We investigated the clinical and prognostic significance of both serum and transcript DKK1 and its functional roles in human hepatocellular carcinoma (HCC). METHODS: We evaluated the expression level of DKK1 in both tissue and serum samples from patients with HCC using GeneChip microarray and real-time-quantitative PCR and sandwich ELISA system respectively. The clinicopathological and prognostic significance of serum and tissue DKK1 levels was examined. Functional characterization of DKK1 with regard to cell migration, invasion and tumour growth was performed. RESULTS: Both DKK1 transcript and serum protein were upregulated in a stepwise manner in human HCCs. Its transcript levels were associated with more aggressive tumour behaviour, in terms of venous invasion (P = 0.003), advanced tumour stage (P = 0.003). DKK1 transcript correlated with shorter overall (P = 0.006) and disease-free survival (P = 0.012), and higher serum DKK1 levels correlated with shorter disease-free survival (P = 0.046). Knockdown of DKK1 significantly reduced both migratory and invasive abilities of HCC cells, whereas overexpression of DKK1 enhanced the tumour formation efficiency and tumour growth in vivo. CONCLUSIONS: Serum and tissue DKK1 levels increased in a stepwise manner in multistep hepatocarcinogenesis and had prognostic significance. DKK1 plays a functional role in cell migration, invasion and tumour growth.

The gene expression of adrenomedullin, calcitonin-receptor-like receptor and receptor activity modifying proteins (RAMPs) in CCl4-induced rat liver cirrhosis.

This study was undertaken to determine AM expression in carbon tetrachloride (CCl4)-induced liver cirrhosis developed with peritoneal ascites. Sprague-Dawley rats received subcutaneous injections of CCl4 twice weekly in olive oil (1:1, 0.3 ml per kg body weight) for 6 or 12 weeks until ascites developed, or saline in olive oil as control. At 6 weeks, fibrosis developed and at 12 weeks cirrhosis developed with ascites formation. At both 6 and 12 weeks, increases in plasma renin and AM were evident, as was the gene expression of AM. At 12 weeks after CCl4 injection, the gene expression of calcitonin-like-receptor (CRLR) and receptor activity modifying proteins (RAMP1, RAMP2 and RAMP3) were all elevated when compared to the control. The results suggest that liver cirrhosis increases mRNA expressions of AM, CRLR and RAMP1, RAMP2 and RAMP3 and that the increase in AM gene expression precedes the development of cirrhosis. The increase in AM synthesis as reflected by an increase in AM gene expression, together with a lack of increase in AM peptide at both 6 and 12 weeks may suggest an elevation of AM release. Given the potent vasodilatory action of AM, the increase in the synthesis and release of AM in the cirrhotic liver may also contribute to peripheral vasodilatation in liver cirrhosis.

CDX2 co-localizes with liver-intestine cadherin in intestinal metaplasia and adenocarcinoma of the stomach.

CDX2 and liver-intestine (LI)-cadherin are intestine-specific markers and both are physiologically expressed in the small intestine and colon. Recent studies have demonstrated that CDX2 regulates LI-cadherin gene (CDH17) expression in colorectal cancer. The present study investigated the relationship of CDX2 and LI-cadherin expression in gastric cancer. One hundred and nine pairs of tumour and non-cancerous gastric mucosa were collected from gastrectomy specimens. Protein expression levels of CDX2 and LI-cadherin were determined by immunohistochemical staining. Semi-quantitative RT-PCR showed that the mRNAs of both CDX2 and CDH17 were highly expressed in tumour compared with non-cancerous mucosa. Overexpression of CDX2 was significantly associated with CDH17 in gastric adenocarcinoma. Furthermore, the expression of CDX2 and LI-cadherin proteins was strongly coupled in intestinal metaplasia. In conclusion, overexpression of CDH17 is significantly associated with CDX2.

Effect of surface oxyfluorination on the dyeability of polyethylene film.

The effect of surface oxyfluorination on low-density polyethylene (LDPE) film was studied in terms of surface functionality and surface energetics of the film surfaces, which can be attributed to improvement of the dyeability. The growth of functional groups and surface free energy was confirmed by FTIR-ATR, XPS, and contact angle methods. As a result, the total surface free energy was increased with oxyfluorination time, as a progressive increase of the polar component together with a small decrease of the dispersive component of surface free energy. From the dyeability test using the Kubelka-Munk equation, it was found that the oxyfluorination treatment plays an important role in the growth of oxygen-containing functional groups of LDPE film, resulting in improving the dyeability with a basic dyeing agent. A direct linear relationship is shown between the specific component of surface free energy and the K/S value for this work.

[Sandwiched osmotic pump tablet for controlled release of water-insoluble drug].

AIM: To study sandwiched osmotic pump tablet for delivering water-insoluble drug for 24 hours. METHODS: Sandwiched osmotic pump tablet was prepared using nifedipine as the model drug. The effects of various formulation variables and orifice size on drug release were studied. The mechanical properties of cellulose acetate membrane were also investigated. RESULTS: Polyethylene oxide of drug layer and potassium chloride of push layer showed marked positive effects on drug release. In the range of 0.50 mm to 1.40 mm, orifice size hardly affects drug release. Cellulose acetate membrane is strong enough to assure the integrity of osmotic pump tablet and could sustain an internal pressure ranging from 0.34 MPa to 2.85 MPa. CONCLUSION: Sandwiched osmotic pump tablet can deliver water-insoluble drug constantly for 24 hours. Release media and agitation rate scarcely affect drug release. Compared with the commercialized push-pull osmotic pump tablet, sandwiched osmotic pump tablet is easy in preparation with exempting identification of drug layer before drilling.

Deregulation of E-cadherin-catenin complex in precancerous lesions of gastric adenocarcinoma.

BACKGROUND AND AIM: Decrease in expression of the E-cadherin-catenin complex is an important element in gastric carcinogenesis. However, the expression of the complex in gastric precancerous lesions has not been well studied. The present study aimed to examine the serial change in expression of E-cadherin-catenin complex in the precancerous lesions of gastric cancer patients. METHODS: Gastrectomy specimens of 40 patients with gastric cancer were retrieved. Areas with chronic gastritis, atrophic gastritis, intestinal metaplasia and adenocarcinoma were identified and immunostained for alpha-catenin, beta-catenin and E-cadherin. The results were scored semiquantitatively by two independent pathologists using a validated scoring system. RESULTS: A significant decrease in score was observed in 5% (1/22) of alpha-catenin, 0% (0/22) of beta-catenin and 9% (2/22) of E-cadherin in chronic atrophic gastritis patients, and in 28% (5/18) of alpha-catenin, 67% (10/15) of beta-catenin and 57% (8/14) of E-cadherin in intestinal metaplasia patients. The scoring of alpha-catenin, beta-catenin and E-cadherin correlated with each other. Forty-three percent of patients had concordant changes of scores along the gastritis-adenocarcinoma sequence. There was no association between Helicobacter pylori status and E-cadherin-catenin complex expression. CONCLUSION: Deregulation of the E-cadherin-catenin complex was observed in the majority of precancerous lesions in patients with gastric adenocarcinoma, which has potential diagnostic and therapeutic implications.

[Unitary-core osmotic pump tablet for controlled release of water-insoluble drug].

AIM: To study unitary-core osmotic pump tablet for delivering water-insoluble drug for 24 hours. METHODS: Unitary-core osmotic pump tablet was prepared using nifedipine as the model drug. The effects of various core formulation variables on drug release were studied. RESULTS: Polyethylene oxide and potassium chloride have comparable positive effects on drug release, whereas, nifedipine has markedly negative effect on drug release. CONCLUSION: Unitary-core osmotic pump tablet is very easy in preparation and it can deliver water-insoluble drug in substantially constant rate for 24 hours.

Sandwiched osmotic pump tablet for controlled release of water-insoluble drug / 药学学报

<p><b>AIM</b>To study sandwiched osmotic pump tablet for delivering water-insoluble drug for 24 hours.</p><p><b>METHODS</b>Sandwiched osmotic pump tablet was prepared using nifedipine as the model drug. The effects of various formulation variables and orifice size on drug release were studied. The mechanical properties of cellulose acetate membrane were also investigated.</p><p><b>RESULTS</b>Polyethylene oxide of drug layer and potassium chloride of push layer showed marked positive effects on drug release. In the range of 0.50 mm to 1.40 mm, orifice size hardly affects drug release. Cellulose acetate membrane is strong enough to assure the integrity of osmotic pump tablet and could sustain an internal pressure ranging from 0.34 MPa to 2.85 MPa.</p><p><b>CONCLUSION</b>Sandwiched osmotic pump tablet can deliver water-insoluble drug constantly for 24 hours. Release media and agitation rate scarcely affect drug release. Compared with the commercialized push-pull osmotic pump tablet, sandwiched osmotic pump tablet is easy in preparation with exempting identification of drug layer before drilling.</p>

Unitary-core osmotic pump tablet for controlled release of water-insoluble drug / 药学学报

<p><b>AIM</b>To study unitary-core osmotic pump tablet for delivering water-insoluble drug for 24 hours.</p><p><b>METHODS</b>Unitary-core osmotic pump tablet was prepared using nifedipine as the model drug. The effects of various core formulation variables on drug release were studied.</p><p><b>RESULTS</b>Polyethylene oxide and potassium chloride have comparable positive effects on drug release, whereas, nifedipine has markedly negative effect on drug release.</p><p><b>CONCLUSION</b>Unitary-core osmotic pump tablet is very easy in preparation and it can deliver water-insoluble drug in substantially constant rate for 24 hours.</p>