Activated Protein C Protects against Murine Contact Dermatitis by Suppressing Protease-Activated Receptor 2

Atopic dermatitis (AD) is a chronic inflammatory skin disease associated with excessive inflammation and defective skin barrier function. Activated protein C (APC) is a natural anticoagulant with anti-inflammatory and barrier protective functions. However, the effect of APC on AD and its engagement with protease activated receptor (PAR)1 and PAR2 are unknown. Methods: Contact hypersensitivity (CHS), a model for human AD, was induced in PAR1 knockout (KO), PAR2KO and matched wild type (WT) mice using 2,4-dinitrofluorobenzene (DNFB). Recombinant human APC was administered into these mice as preventative or therapeutic treatment. The effect of APC and PAR1KO or PARKO on CHS was assessed via measurement of ear thickness, skin histologic changes, inflammatory cytokine levels, Th cell phenotypes and keratinocyte function. Results: Compared to WT, PAR2KO but not PAR1KO mice displayed less severe CHS when assessed by ear thickness; PAR1KO CHS skin had less mast cells, lower levels of IFN-γ, IL-4, IL-17 and IL-22, and higher levels of IL-1β, IL-6 and TGF-β1, whereas PAR2KO CHS skin only contained lower levels of IL-22 and IgE. Both PAR1KO and PAR2KO spleen cells had less Th1/Th17/Th22/Treg cells. In normal skin, PAR1 was present at the stratum granulosum and spinosum, whereas PAR2 at the upper layers of the epidermis. In CHS, however, the expression of PAR1 and PAR2 were increased and spread to the whole epidermis. In vitro, compared to WT cells, PAR1KO keratinocytes grew much slower, had a lower survival rate and higher para permeability, while PAR2KO cells grew faster, were resistant to apoptosis and para permeability. APC inhibited CHS as a therapeutic but not as a preventative treatment only in WT and PAR1KO mice. APC therapy reduced skin inflammation, suppressed epidermal PAR2 expression, promoted keratinocyte growth, survival, and barrier function in both WT and PAR1KO cells, but not in PAR2KO cells. Conclusions: APC therapy can mitigate CHS. Although APC acts through both PAR1 and PAR2 to regulate Th and mast cells, suppression of clinical disease in mice is achieved mainly via inhibition of PAR2 alone. Thus, APC may confer broad therapeutic benefits as a disease-modifying treatment for AD.

Combined mRNAs and clinical factors model on predicting prognosis in patients with triple-negative breast cancer

Objective Triple-negative breast cancer (TNBC) is aggressive cancer usually diagnosed in young women with no effective prognosis prediction model to use. The present study was performed to develop a useful prognostic model for predicting overall survival (OS) for TNBC patients. Methods The Cancer Genome Atlas (TCGA) and Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) databases were used as training and validation data sets, respectively, in which the gene expression levels and clinical prognostic information of TNBC were collected. Differentially expressed genes (DEGs) between TNBC and non-TNBC (NTNBC) were identified with the thresholds of false discovery rate < 0.05 and |log2 Fold Change| > 1. DEGs in AmiGO2 and the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were retained for further study. Univariate, multivariate Cox, and logistic regression analysis were conducted for detecting DEG signature with the threshold of log-rank P < 0.05. The prognosis models of mRNA signature, clinical factors were constructed and compared. Results One five-DEG signature, including CHST4, COCH, CST9, SOX11, and TDGF1 was identified in DEG prognosis model. Stratified analysis showed that the patients aged over 60, with higher pathologic stage (III-IV) and recurrence induced a significantly lower survival rate than those aged below 60, lower pathologic stage and without recurrence. Compared with patients with low-risk scores, those presented high-risk scores demonstrated significantly lower survival rate in the subgroup aged over 60 [HR = 3.780 (1.801–7.933), P < 0.0001]. For patients who obtained a higher pathologic stage and recurrence, high-risk scores were correlated with a significantly lower survival rate than patients with low-risk scores. The five-mRNA signature combined with clinical model (AUC = 0.950) predicted better than single clinical model (AUC = 0.795) or five-mRNA signature model (AUC = 0.823). Conclusion Our present study identified a prognostic prediction model (combined with five-mRNA signature and clinical factors) for TNBC patients receiving immunotherapy, which will benefit future research and clinical therapies.

Clinical Profile of 24 AIDS Patients with Cryptococcal Meningitis in the HAART Era: A Report from an Infectious Diseases Tertiary Hospital in Western Romania

Management of cryptococcal infections among patients suffering from acquired immunodeficiency syndrome (AIDS) represents a medical challenge. This retrospective study aims to describe the disease management and outcomes among 24 AIDS patients who suffered from Cryptococcus neoformans meningitis. The parameters evaluated from our patients’ database records include epidemiological data, clinical manifestations, biochemical and microbiological analysis of patients’ cerebrospinal fluid (CSF), treatment profiles, and disease outcomes. All patients included in the study had a lymphocyte count of less than 200 CD4/mm3. Of the 24 patients included in this study, five had been diagnosed with HIV infection since childhood, after receiving HIV-infected blood transfusions. The most prominent symptom was fatigue in 62.5% of patients, followed by nausea/vomiting and headache. Seven patients had liver cirrhosis due to hepatitis B virus (HBV) or hepatitis C virus (HCV) infection, while Kaposi sarcoma and cerebral toxoplasmosis were found in two patients. Six out of 24 patients died due to bacterial sepsis and acute respiratory distress syndrome (ARDS). High intracranial pressure was the strongest predictive factor for mortality (OR = 2.9), followed by ARDS (OR = 1.8), seizures at disease onset (OR = 1.4), and diabetes mellitus (OR = 1.2). Interestingly, patients younger than 40 years old had a significantly lower survival rate than that of the older patients. Before developing Cryptococcal meningitis, all patients had low adherence to the early ART treatment scheme and skipped the follow-up visits. All patients received a combination of amphotericin B and flucytosine as induction therapy, adding fluconazole for maintenance. Simultaneously, AIDS HAART was initiated at diagnosis of the cryptococcal infection. A combined regimen of antifungals and highly active antiretroviral therapy showed improved patient recovery with minor side effects.

“The obesity paradox” in patients with atrial fibrillation according to the results of the REKUR-AF study

Introduction: to evaluate the effect of excess body weight (EBW) and obesity on the survival of patients with atrial fibrillation (AF) in the REKUR-AF study. Materials and methods: A subanalysis of patients with AF included in the REKUR-AF (382 people) study was performed. Survival rates were analyzed in three patient groups: patients with normal body mass index (BMI), EBW and obesity. Then the nature and significance of the influence of the studied factor on the prognosis in patients with AF were assessed. Results and discussion: Patients with AF and normal BMI were significantly older than those with obesity and EBW (p&lt;0.001 and p=0.021, respectively). Among obese patients, hypertension was significantly (p=0.0015) more common (93.9%) compared to the group of patients with a normal BMI level (80.5%). The frequency of type 2 diabetes in obese patients prevailed over the same indicator in the groups of people with normal BMI and EBW (p=0.007 and p=0.020, respectively). The analysis of the survival rate of patients with AF depending on the BMI level showed that this factor had a significant impact (p=0.013) on the prognosis. The group of individuals with a normal BMI level had a significantly lower survival rate than the cohort of patients with EBW (p=0.011) and OB (p=0.025). The final model for Cox regression analysis (χ2=53.06, p&lt;0.001) included the following factors: age, form of AF, BMI, presence/absence of type 2 diabetes, presence/absence of previous hospitalizations, presence/absence of hypertension, and presence/absence of oral anticoagulant (OAC) intake. Conclusion: The obtained results do not make it possible to unambiguously interpret obesity as a predictor of a positive outcome in this category of individuals.

Estimating abundance of a recovering transboundary brown bear population with capture-recapture models

Abundance of small populations of large mammals may be assessed using complete counts of the different individuals detected over a time period, so-called minimum detected size (MDS). However, as population is growing larger and its distribution is expanding wider, the risk of under-estimating population size using MDS is increasing sharply due to the rarely fulfilled assumption of perfect detection of all individuals of the population, and as a result, the need to report uncertainty in population size estimates becomes crucial. We addressed these issues within the framework of the monitoring of the critically endangered Pyrenean brown bear population that was on the edge of extinction in the mid-1990s with only five individuals remaining, but was reinforced by 11 bears originated from Slovenia since then. We used Pollock's closed robust design (PCRD) capture-recapture models applied to the cross-border non-invasive sampling data from France, Spain and Andorra to provide the first published annual abundance estimates of the Pyrenean brown bear population and its trends over time. Annual population size increased and displayed a fivefold rise between 2008 and 2020, reaching > 60 individuals in 2020. Detection heterogeneity among individuals may stem from intraspecific home range size disparities making it more likely to find signs of individuals who move more. We found a lower survival rate in cubs than in adults and subadults, since the formers suffer from more mortality risks (such as infanticides, predations, mother death or abandonments) than the latters. Our study provides evidence that the PCRD capture-recapture modelling approach can provide reliable estimates of the size of and trend in large mammal populations, while minimizing bias due to inter-individual heterogeneity in detection probabilities and allowing the quantification of sampling uncertainty surrounding these estimates. Such information is vital for informing management decision-making and assessing population conservation status.

Impact of Comorbidities and COVID-19 on the Survival Rate of Geriatric Patients Receiving Anesthesia Services: A Prospective Cohort Study in Indonesia

Background As life expectancy increases, the worldwide population aged 60 years and older increases year by year. Consequently, more older people receive medical attention, especially those who undergo surgery. In addition, the COVID-19 pandemic has had a global impact on elderly patients, especially those undergoing surgery. This study aims to describe the characteristics and analyze the survival rate of elderly patients who receive anesthesia services, especially those with comorbidities and COVID-19. Methods A prospective cohort study at 14 central hospitals in Indonesia analyzed 1621 elderly patients (67.1 ± 6.2 years old). The variables that were recorded included patient characteristics, comorbidities, the COVID-19 status, and the survival rate, including 30-day mortality. Results The 30-day mortality was 4.4%. The most comorbidity was hypertension (30.0). Patients with a Charlson's Comorbidity Index Score of 3-4 had a higher death rate (15.3%). The highest mortality rates were in the patients who had dementia, rheumatologic disease, liver disease, previous myocardial infarction, and diabetes with chronic complications as comorbidities. The percent of patients with COVID-19 who died was 26.6%. Patients with several comorbidities and COVID-19 had a lower survival rate than those without (log-rank p<0.05) Conclusion Approximately four in ten elderly patients receiving anesthesia died, and the percent increased when the patients had comorbidities and COVID-19.

Decreased ARID1A Expression Associates with Tumor Progression and Adverse Prognosis in Renal Cell Carcinoma

In this study, we aimed to evaluate association of ARID1A (AT-rich interacting domain-containing protein 1A) mutation and protein expression with clinicopathology and prognosis of renal cell carcinoma (RCC). Genomic Data Commons (GDC) showed ARID1A was one of the top-ten mutated genes found in kidney cancers and its mutations were found along its sequence. Interestingly, patients with ARID1A mutations had significantly lower survival rate (38%; n=68) comparing to the non-mutated cases (58%; n=192). The results from OSkirc web tool revealed that patients with low expression of ARID1A had significantly shorter overall survival and disease specific survival than those with high ARID1A expression. Immunohistochemistry revealed markedly decreased ARID1A expression in the RCC tissues (n=26), particularly in clear cell RCC (ccRCC) and chromophobe RCC (chRCC). Negative to weak ARID1A expression was significantly associated with ccRCC (grade II) and chRCC subtypes, presence of comorbidity, and low eGFR levels. Finally, ARID1A protein was undetectable in 3/11 cases with ccRCC (grade II) and 2/6 chRCC cases, all of which had metastasis 1−50 months after surgical removal. In conclusion, decreased ARID1A expression is associated with the poor prognosis and metastasis of RCC and thus may serve as the prognostic marker of RCC, particularly ccRCC and chRCC subtypes.

Abstract 10476: Patient Characteristics and Survival Outcomes of Cardiac Arrest in the Cardiac Catheterization Laboratory: Insights from Get with the Guidelines-Resuscitation Registry

Background: The characteristics and outcomes of in-hospital cardiac arrest (IHCA) in the cardiac catheterization laboratory (CCL) have not been well-described. We compared the characteristics and outcomes of patients with an IHCA in the CCL versus those in the operating room (OR) and the intensive care unit (ICU). Methods: Within the American Heart Association’s Get With the Guidelines-Resuscitation® registry, we identified patients 18 years of age or older with an IHCA in the CCL, OR, or ICU between 2000 and 2019. We compared rates of survival to discharge for patients in the CCL, OR, and ICU. Additionally, we examined predictors of survival to discharge for patients with IHCA in the CCL. Results: There were 6866, 5181, and 181,832 patients with an IHCA in the CCL, OR, and ICU, respectively. Patients with IHCAs in the CCL were more likely to have a shockable cardiac arrest rhythm as compared with those in the OR and ICU. Overall, 2614 (38.1%) patients with IHCA in the CCL survived to discharge, as compared with 30,833 (16.9%) from the ICU and 2096 (40.5%) from the OR. After adjustment for 27 patient and cardiac arrest factors, patients with IHCA in CCL were more likely to survive to discharge as compared with those with IHCA from the ICU (odds ratio, 1.37 [95% CI: 1.29-1.46], p<0.001). In contrast, they were less likely to survive to discharge as compared with those with IHCA in the OR (odds ratio, 0.81 [95% CI: 0.69-0.94], p=0.006). Predictors of survival to discharge in patients with IHCA in the CCL included white race, pulseless ventricular tachycardia/fibrillation, and IHCA during normal hours and on weekdays, while having myocardial infarction during this or prior hospitalization was associated with less survival to discharge. (Table). Conclusion: IHCA in the CCL is not uncommon and has a lower survival rate as compared with IHCA in other procedural areas such as the OR. The reasons for this difference deserve further study given that response to IHCAs in both settings should be similar.

VDR Signaling via the Enzyme NAT2 Inhibits Colorectal Cancer Progression

Recent epidemiological and preclinical evidence indicates that vitamin D3 inhibits colorectal cancer (CRC) progression, but the mechanism has not been completely elucidated. This study was designed to determine the protective effects of vitamin D3 and identify crucial targets and regulatory mechanisms in CRC. First, we confirmed that 1,25(OH)2D3, the active form of vitamin D3, suppressed the aggressive phenotype of CRC in vitro and in vivo. Based on a network pharmacological analysis, N-acetyltransferase 2 (NAT2) was identified as a potential target of vitamin D3 against CRC. Clinical data of CRC patients from our hospital and bioinformatics analysis by online databases indicated that NAT2 was downregulated in CRC specimens and that the lower expression of NAT2 was correlated with a higher metastasis risk and lower survival rate of CRC patients. Furthermore, we found that NAT2 suppressed the proliferation and migration capacity of CRC cells, and the JAK1/STAT3 signaling pathway might be the underlying mechanism. Moreover, Western blot and immunofluorescence staining assays demonstrated that 1,25(OH)2D3 promoted NAT2 expression, and the chromatin immunoprecipitation assay indicated that the vitamin D receptor (VDR) transcriptionally regulated NAT2. These findings expand the potential uses of vitamin D3 against CRC and introduce VDR signaling via the enzyme NAT2 as a potential diagnostic and therapeutic target for CRC.

The expression and significance of 8-hydroxydeoxyguanosine in breast cancer patients’ blood, urine and cancer tissue

Objective: To explore the expression and clinic significance of 8-OHdG in breast cancer. Methods: Pre-operative serum 8-OHdG levels were detected with an enzyme-linked immunosorbent assay in a well-defined series of 173 breast cancer patients. 8-OHdG expression in cancer cells from 150 of these patients was examined by immunohistochemistry. The HPLC-ECD method is used to determine 8-OHdG concentration in urine. Results: The serum 8-OHdG levels and immunohistochemical 8-OHdG expression were in concordance with each other (P < 0.05, r = 0.163). Breast cancer patients with negative 8-OHdG immunostaining show lower survival rate according to the multivariate analysis (P < 0.01). This observation was even more remarkable in ductal carcinomas (n = 140) patients (P < 0.001). A low serum 8-OHdG level was associated statistically significantly with lymphatic vessel invasion and a positive lymph node status. Comparison of 8-OHdG concentration in urine of breast cancer patients and healthy women was statistical significance (P < 0.01). Conclusion: Low serum 8-OHdG levels and a low immunohistochemical 8-OHdG expression were associated with an aggressive breast cancer phenotype. In addition, negative 8-OHdG immunostaining was an independent prognostic factor for breast cancer-specific death in breast carcinoma patients. Using 8-OHdG concentration in urine to predict DNA damage resulting from breast cancer can provide good biological indicators for detecting harm in early breast cancer.