Radiosensitizing effect of a novel CTSS inhibitor by enhancing BRCA1 protein stability in triple-negative breast cancer cells.

Triple-negative breast cancer (TNBC) patients harboring wild-type breast cancer susceptibility gene 1 (BRCA1) account for most TNBC patients but lack adequate targeted therapeutic options. Although radiotherapy (RT) is the primary treatment modality for TNBC patients, radioresistance is one of the major challenges. RT-induced increase in cathepsin S (CTSS) causes radioresistance through suppressing BRCA1-mediated apoptosis of tumor cells, which was induced by CTSS-mediated degradation of BRCA1. Targeting CTSS may provide a novel therapeutic opportunity for TNBC patients. Publicly available data and human tissue microarray slides were analyzed to investigate the relationship between CTSS and BRCA1 in breast cancer patients. A CTSS enzyme assay and in silico docking analysis were conducted to identify a novel CTSS inhibitor. RO5461111 was used first to confirm the concept of targeting CTSS for radiosensitizing effects. The MDA-MB-231 TNBC cell line was used for in vitro and in vivo assays. Western blotting, promoter assay, cell death assay, clonogenic survival assay, and immunohistochemistry staining were conducted to evaluate novel CTSS inhibitors. CTSS inhibitors were further evaluated for their additional benefit of inhibiting cell migration. A novel CTSS inhibitor, TS-24, increased BRCA1 protein levels and showed radiosensitization in TNBC cells with wild-type BRCA1 and in vivo in a TNBC xenograft mouse model. These effects were attributed by BRCA1-mediated apoptosis facilitated by TS-24. Furthermore, TS-24 demonstrated the additional effect of inhibiting cell migration. Our study suggests that employing CTSS inhibitors for the functional restoration of BRCA1 to enhance RT-induced apoptosis may provide a novel therapeutic opportunity for TNBC patients harboring wild-type BRCA1.

Triptolide alleviates cerebral ischemia/reperfusion injury via regulating the Fractalkine/CX3CR1 signaling pathway.

PURPOSE: To evaluate the potential efficacy of Triptolide (TP) on cerebral ischemia/reperfusion injury (CIRI) and to uncover the underlying mechanism through which TP regulates CIRI. METHODS: We constructed a middle cerebral artery occlusion/reperfusion (MCAO/R) mouse model to simulate CIRI, and established a lipopolysaccharide (LPS)-stimulated BV-2 cell model to mimic the inflammatory state during CIRI. The neurological deficits score (NS) of mice were measured for assessment of neurologic functions. Both the severity of cerebral infarction and the apoptosis level in mouse brain tissues or cells were respectively evaluated using corresponding techniques. The expression levels of Ionized calcium binding adapter molecule 1 (IBA-1), Inductible Nitric Oxide Synthase (iNOS), Arginase 1 (Arg-1), Tumor necrosis factor-α (TNF-α), Interleukin 1ß (IL-1ß), Cysteine histoproteinase S (CTSS), Fractalkine, chemokine C-X3-C motif receptor 1 (CX3CR1), BCL-2-associated X protein (BAX), and antiapoptotic proteins (Bcl-2) were detected using immunofluorescence, qRT-PCR as well as Western blot, respectively. RESULTS: Relative to the Sham group, treatment with TP attenuated the increased NS, infarct area and apoptosis levels observed in MCAO/R mice. Upregulated expression levels of IBA-1, iNOS, Arg-1, TNF-α and IL-1ß were found in MCAO/R mice, while TP suppressed iNOS, TNF-α and IL-1ß expression, and enhanced Arg-1 expression in both MCAO/R mice and LPS-stimulated BV-2 cells. Besides, TP inhibited the CTSS/Fractalkine/CX3CR1 pathway activation in both MCAO/R mice and LPS-induced BV-2 cells, while overexpression of CTSS reversed such effect. Co-culturing HT-22 cells with TP+LPS-treated BV-2 cells led to enhanced cell viability and decreased apoptosis levels. However, overexpression of CTSS further aggravated HT-22 cell injury. CONCLUSION: TP inhibits not only microglia polarization towards the M1 phenotype by suppressing the CTSS/Fractalkine/CX3CR1 pathway activation, but also HT-22 apoptosis by crosstalk with BV-2 cells, thereby ameliorating CIRI. These findings reveal a novel mechanism of TP in improving CIRI, and offer potential implications for addressing the preventive and therapeutic strategies of CIRI.

Diagnostic Yield of Stereotactic Brain Biopsy in a Sub-Saharan Tertiary Center: A Comprehensive 10-Year Retrospective Analysis.

BACKGROUND: Stereotactic brain biopsy is a crucial minimally invasive surgical technique leveraged to obtain tissue specimens from deep-seated intracranial lesions, offering a safer alternative to open craniotomy for patients who cannot tolerate the latter. Despite its effectiveness, the diagnostic yield varies across different centers and has not been widely studied in Sub-Saharan Africa. METHODS: A single-center retrospective analysis was conducted on 67 consecutive stereotactic brain biopsy procedures carried out by experienced neurosurgeons between January 2012 and December 2022 at a tertiary center in Sub-Saharan Africa. Preoperative clinical status, biopsy type, postoperative complication rate, and histological diagnosis were meticulously analyzed. Factors associated with negative biopsy results were identified using IBM Statistical Package for the Social Sciences SPSS version for Mac, with Fisher exact test employed to detect differences in patient characteristics. Statistical significance was pegged at P < 0.05. RESULTS: The overall diagnostic yield rate was 67%. Major contributors to negative biopsy outcomes were superficial location of the lesion, lesion size less than 10 cc, and the use of the Cape Town Stereotactic System. Enhanced yield rates of up to 93% were realized through the application of magnetic resonance imaging-based images, Stealth Station 7, and frozen section analysis. No correlation was observed between the number of cores obtained and the yield rate. Procedure complications were negligible, and no procedure-related mortality was recorded. CONCLUSIONS: The diagnostic yield rate from our study was somewhat lower than previously reported in contemporary literature, primarily attributed to the differing definitions of diagnostic yield, the dominant use of the older framed Cape Town Stereotactic System, computed tomography-based imaging, and the absence of intraoperative frozen section. Nevertheless, biopsies conducted using the frameless system were comparable with studies from other global regions. Our findings reaffirm that stereotactic brain biopsy when complemented with magnetic resonance imaging-based imaging, frameless stereotactic systems and intraoperative frozen section is a safe, effective, and reliable method for obtaining histological diagnosis.

Effect of CTSS non-synonymous mutations on litter size in Qianbei Ma goats.

Cathepsin S (CTSS) is a member of the cysteine protease family closely related to reproductive regulation in goats. However, its effect on litter size in goats remains unclear. In this study, the relationship between CTSS gene polymorphisms and litter size was revealed by analyzing the DNA sequence and mRNA expression of CTSS in the gonadal axis of Qianbei Ma goats. In addition, bioinformatics methods were used to evaluate the effect of non-synonymous mutations on CTSS protein structure and function. CTSS was expressed in all parts of the gonadal axis of Qianbei Ma goats, with the highest expression in the uterus in the multi-lamb group and in the fallopian tube in the single-lamb group. The sequencing results showed that four SNPs in CTSS, including g.7413C → T, g.8816A → T, g.9191 T → G and g.10193G → A, were significantly correlated with litter size (p < 0.05). All four analyzed mutation sites were in strong linkage disequilibrium (r2 > 0.33, D' > 0.70). Additionally, the haplotype Hap1/2 had a significantly higher frequency than the other haplotypes (p < 0.05). g.7413C → T and g.8816A → T were non-synonymous mutations. The g.7413C → T mutation resulted in the substitution of serine 161 of the CTSS protein with phenylalanine (p.S161F), and the g.8816A → T mutation resulted in the substitution of aspartate 219 with tyrosine (p.N219Y). p.S161F was highly conserved across 13 species and that p.N219Y was relatively conserved in cloven-hoofed species. Mutations at two sites changed the local conformation of the CTSS protein, reduced its stability, and affected its function and goat breed evolution. These findings confirm that CTSS affects the lambing traits of goats and provide a theoretical basis for the regulatory mechanism of CTSS in affecting litter size.

Diagnostic Performance Between Chest CT Severity Score and Initial Reverse Transcription-Polymerase Chain Reaction (RT-PCR) Cycle Values in COVID-19 Patients and Their Relation With the Clinical Status of Patients.

INTRODUCTION: Reverse transcription-polymerase chain reaction (RT-PCR) is used as a standard test for the diagnosis of SARS-CoV-2 viral RNA from nasopharyngeal aspirates. However, this method lacks sensitivity and cannot assess disease severity. A CT scan of the thorax provides a CT severity score (CT-SS), which depicts lung involvement and disease severity. This study aims to investigate the diagnostic value of chest CT compared with RT-PCR cycle threshold (Ct) values in COVID-19 and relate it clinically with the disease severity of patients. METHODS: This retrospective observational study was conducted in a tertiary center from April 2021 to March 2022. We included 511 patients who had tested RT-PCR positive for COVID-19, were hospitalized, and had undergone high-resolution CT (HRCT) thorax. Data was collected from patient records regarding name, age, sex, admission data, baseline investigations including Ct value, management, and outcome. HRCT was reviewed to assess lung involvement and calculate CT-SS. Data was analyzed using SPSS Statistics version 25 (IBM Corp. Released 2017. IBM SPSS Statistics for Windows, Version 25.0. Armonk, NY: IBM Corp.). RESULT: The mean age of patients was 50.4 ± 13.7 years, and the majority (67.5%) were male. Gender-wise, there was no difference in RT-PCR cycle threshold (Ct) values; however, CT-SS was significantly higher in males (17.5 ± 4.8 vs.10.5 ± 6.6, t=-13.6, p<0.0001). ICU admission was needed for 34.8% of patients, and they had a significantly lower Ct value (21.7 ± 3.3 vs. 22.8 ± 3.7, t=21.10, p<0.0001) and higher CT-SS (16.3 ± 4.5 vs. 6.7 ± 5.1, t=-3.32, p=0.001). CONCLUSION: Ct values could not differentiate between moderate and severe patients. CT-SS was not related to the viral load at admission. Patients who succumbed had significantly lower Ct values and higher CT-SS.

Clinical profile of hospitalised moderate category COVID-19 patients: Short study from a Tertiary Care Centre in Delhi.

Background: The clinical profile of hospitalized moderate-category COVID-19 patients has been understudied globally and in India. Aim: The present study was conducted to study the clinical profile and assess the proportions of patients who progressed to severe disease and its predictors among moderate COVID-19 patients. Materials and Methods: In this single-center observational study, 100 moderate-category COVID-19 patients as per Ministry of Health and Family Welfare (MoHFW) criteria of age ≥18 years of either sex, excluding pregnant females from February to November 2021, were studied by analyzing their clinical profiles and assessing Quick Sequential Organ Failure Assessment (qSOFA), National Early Warning Score 2 (NEWS-2), and chest computed-tomography severity score (CTSS) to predict progression to severe disease. Severe disease was defined as per MoHFW criteria. Results: Out of 100 moderate-category COVID-19 patients, progression to severe disease was seen in 11 patients (11%), among which eight patients had expired, three patients were discharged, and the rest of the 89 patients (89%) who did not progress to severe disease were discharged. A higher age (62.2± 19.5 vs 54.8 ± 14.6 years), along with multivariate analysis revealing male sex (1.25 times), chronic kidney disease (2.86 times), leukocytosis (6.10 times), thrombocytopenia (1.04 times), anemia (9.3 times), a higher qSOFA score (3.6 times), and a higher NEWS-2 score on admission (1.56 times) had higher odds of progression to severe disease. A significant correlation (P < .05) of qSOFA score with serum LDH, ferritin, and hs-CRP levels; CT severity score with the serum ferritin, IL-6, and LDH levels; and NEWS-2 with serum LDH, hs-CRP, and ferritin levels were found. Moreover, the NEWS-2 score was found slightly better than qSOFA on receiver operating characteristic (ROC) curve analysis, with an area under the curve of 85.8% and 83.2%, respectively, predicting progression to severe disease. Conclusion: Our study revealed male gender, chronic kidney disease, leukocytosis, anemia, thrombocytopenia, a higher qSOFA and NEWS-2 score on admission, and further, NEWS-2 score better than qSOFA on ROC curve analysis, with an area under the curve of 85.8% and 83.2%, respectively, in predicting severe disease among hospitalized moderate COVID-19 patients.

Cathepsin S inhibition in dendritic cells prevents Th17 cell differentiation in perivascular adipose tissues following vascular injury in diabetic rats.

In the context of diabetes mellitus (DM), the circulating cathepsin S (CTSS) level is significantly higher in the cardiovascular disease group. Therefore, this study was designed to investigate the role of CTSS in restenosis following carotid injury in diabetic rats. To induce DM, 60 mg/kg of streptozotocin (STZ) in citrate buffer was injected intraperitoneally into Sprague-Dawley rats. After successful modeling of DM, wire injury of the rat carotid artery was performed, followed by adenovirus transduction. Levels of blood glucose and Th17 cell surface antigens including ROR-γt, IL-17A, IL-17F, IL-22, and IL-23 in perivascular adipose tissues (PVAT) were evaluated. For in vitro analysis, human dendritic cells (DCs) were treated with 5.6-25 mM glucose for 24 h. The morphology of DCs was observed using an optical microscope. CD4+ T cells derived from human peripheral blood mononuclear cells were cocultured with DCs for 5 days. Levels of IL-6, CTSS, ROR-γt, IL-17A, IL-17F, IL-22 and IL-23 were measured. Flow cytometry was conducted to detect DC surface biomarkers (CD1a, CD83, and CD86) and Th17 cell differentiation. The collected DCs presented a treelike shape and were positive for CD1a, CD83, and CD86. Glucose impaired DC viability at the dose of 35 mM. Glucose treatment led to an increase in CTSS and IL-6 expression in DCs. Glucose-treated DCs promoted the differentiation of Th17 cells. CTSS depletion downregulated IL-6 expression and inhibited Th17 cell differentiation in vitro and in vivo. CTSS inhibition in DCs inhibits Th17 cell differentiation in PVAT tissues from diabetic rats following vascular injury.

Predicting Severe COVID-19 Infection on Initial Diagnosis: Comparison and Validation of CT Imaging Triage Tools.

BACKGROUND: Developing a reliable predictive tool of disease severity in COVID-19 infection is important to help triage patients and ensure the appropriate utilization of health-care resources. OBJECTIVE: To develop, validate, and compare three CT scoring systems (CTSS) to predict severe disease on initial diagnosis of COVID-19 infection. METHODS: One hundred and twenty and 80 symptomatic adults with confirmed COVID-19 infection who presented to emergency department were evaluated retrospectively in the primary and validation groups, respectively. All patients had non-contrast CT chest within 48 hours of admission. Three lobarbased CTSS were assessed and compared. The simple lobar system was based on the extent of pulmonary infiltration. Attenuation corrected lobar system (ACL) assigned further weighting factor based on attenuation of pulmonary infiltrates. Attenuation and volume-corrected lobar system incorporated further weighting factor based on proportional lobar volume. The total CT severity score (TSS) was calculated by adding individual lobar scores. The disease severity assessment was based on Chinese National Health Commission guidelines. Disease severity discrimination was assessed by the area under the receiver operating characteristic curve (AUC). RESULTS: The ACL CTSS demonstrated the best predictive and consistent accuracy of disease severity with an AUC of 0.93(95%CI:0.88-0.97) in the primary cohort and 0.97 (95%CI:0.91.5-1) in the validation group. Applying a TSS cut-off value of 9.25, the sensitivities were 96.4% and 100% and the specificities were 75% and 91% in the primary and validation groups, respectively. CONCLUSION: The ACL CTSS showed the highest accuracy and consistency in predicting severe disease on initial diagnosis of COVID-19. This scoring system may provide frontline physicians with a triage tool to guide admission, discharge, and early detection of severe illness.

Identification of a three-gene prognostic signature for radioresistant esophageal squamous cell carcinoma.

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is causing a high mortality rate due to the lack of efficient early prognosis markers and suitable therapeutic regimens. The prognostic role of genes responsible for the acquisition of radioresistance in ESCC has not been fully elucidated. AIM: To establish a prognostic model by studying gene expression patterns pertinent to radioresistance in ESCC patients. METHODS: Datasets were obtained from the Gene Expression Omnibus and The Cancer Genome Atlas databases. The edgeR, a Bioconductor package, was used to analyze mRNA expression between different groups. We screened genes specifically responsible for radioresistance to estimate overall survival. Pearson correlation analysis was performed to confirm whether the expression of those genes correlated with each other. Genes contributing to radioresistance and overall survival were assessed by the multivariate Cox regression model through the calculation of ßi and risk score using the following formula: . RESULTS: We identified three prognostic mRNAs (cathepsin S [CTSS], cluster of differentiation 180 [CD180], and SLP adapter and CSK-interacting membrane protein [SCIMP]) indicative of radioresistance. The expression of the three identified mRNAs was related to each other (r > 0.70 and P < 0.05). As to 1-year and 3-year overall survival prediction, the area under the time-dependent receiver operating characteristic curve of the signature consisting of the three mRNAs was 0.716 and 0.841, respectively. When stratifying patients based on the risk score derived from the signature, the high-risk group exhibited a higher death risk and shorter survival time than the low-risk group (P < 0.0001). Overall survival of the low-risk patients was significantly better than that of the high-risk patients (P = 0.018). CONCLUSION: We have developed a novel three-gene prognostic signature consisting of CTSS, CD180, and SCIMO for ESCC, which may facilitate the prediction of early prognosis of this malignancy.

Cathepsin S inhibitor reduces high-fat-induced adipogenesis, inflammatory infiltration, and hepatic lipid accumulation in obese mice.

Background: Obesity, which results from a caloric intake and energy expenditure imbalance, is highly prevalent worldwide. Cathepsin S (CTSS), which is a cysteine protease, is elevated in obesity and may regulate a variety of physiological processes. This study sought to investigate the functional role of CTSS in obesity. Methods: Mice were administrated 60 mg/kg of RO5444101 in vivo and fed a high-fat diet (HFD) to induce obesity. The weights of the mice fed a normal-chow diet and a HFD were measured. The expression levels of total triglycerides (TG), total cholesterol (TC), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and monocyte chemoattractant protein-1 (MCP-1) were assessed using appropriate corresponding assay kits. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to estimate the messenger ribonucleic acid (mRNA) expression of CTSS in the serum and the release of M1- and M2-type cytokines, and western blot was used to measure the phosphorylated-nuclear factor kappaB (NF-kappaB) p65 and NF-κB p65 proteins. The mRNA and protein expressions of sterol regulatory element-binding protein 1 (SREBP1), fatty acid synthase (FASN), leptin, and adiponectin were also evaluated by RT-qPCR and western blot. Further, hematoxylin and eosin (H&E), immunohistochemical, and red oil O staining were employed to detect the pathological changes of the epididymal white adipose tissue (eWAT), the macrophage infiltration in the eWAT, and lipid accumulation, respectively. Results: We found that CTSS was elevated in the plasma, visceral adipose, and liver tissues of the obese mice. After the administration of 60 mg/kg of RO5444101, the weight of the obese mice decreased, insulin resistance was inhibited, and adipocyte formation was suppressed. The CTSS inhibitor also decreased the level of macrophage infiltration in the eWAT, MCP-1 expression, and the release of M1- and M2-type cytokines in the HFD-induced mice. The CTSS inhibitor appeared to improve the hepatic function parameters and lipid accumulation of the HFD-induced mice. The CTSS inhibitor also appeared to improve the inflammatory damage in the HFD-induced mice. Conclusions: CTSS inhibitor helped to protect against HFD-induced adipogenesis, inflammatory infiltration, and hepatic lipid accumulation in obese mice.

Point-of-care Lung Ultrasound, Lung CT and NEWS to Predict Adverse Outcomes and Mortality in COVID-19 Associated Pneumonia.

Introduction: The appraisal of disease severity and prediction of adverse outcomes using risk stratification tools at early disease stages is crucial to diminish mortality from coronavirus disease 2019 (COVID-19). While lung ultrasound (LUS) as an imaging technique for the diagnosis of lung diseases has recently gained a leading position, data demonstrating that it can predict adverse outcomes related to COVID-19 is scarce. The main aim of this study is therefore to assess the clinical significance of bedside LUS in COVID-19 patients who presented to the emergency department (ED). Methods: Patients with a confirmed diagnosis of SARS-CoV-2 pneumonia admitted to the ED of our hospital between March 2021 and May 2021 and who underwent a 12-zone LUS and a lung computed tomography scan were included prospectively. Logistic regression and Cox proportional hazard models were used to predict adverse events, which was our primary outcome. The secondary outcome was to discover the association of LUS score and computed tomography severity score (CT-SS) with the composite endpoints. Results: We assessed 234 patients [median age 59.0 (46.8-68.0) years; 59.4% M), including 38 (16.2%) in-hospital deaths for any cause related to COVID-19. Higher LUS score and CT-SS was found to be associated with ICU admission, intubation, and mortality. The LUS score predicted mortality risk within each stratum of NEWS. Pairwise analysis demonstrated that after adjusting a base prediction model with LUS score, significantly higher accuracy was observed in predicting both ICU admission (DBA -0.067, P = .011) and in-hospital mortality (DBA -0.086, P = .017). Conclusion: Lung ultrasound can be a practical prediction tool during the course of COVID-19 and can quantify pulmonary involvement in ED settings. It is a powerful predictor of ICU admission, intubation, and mortality and can be used as an alternative for chest computed tomography while monitoring COVID-19-related adverse outcomes.

Comparison of Imaging Severity Between Vaccinated and Unvaccinated COVID-19 Patients: Perspective of an Indian District.

BACKGROUND: Extensive vaccination drives undertaken globally helped in the fight against the coronavirus disease 2019 (COVID-19) pandemic, but different nations adopted different vaccination policies to tackle the disease. The vaccination drive in India began with the administration of two different vaccines: Covishield and Covaxin. We assessed the effect of vaccination status on imaging severity in patients with positive COVID-19 reverse transcription-polymerase chain reaction (RT-PCR)/antigen tests. METHOD: This was a single-center retrospective observation analysis carried out over three months between March 1, 2021, to May 31, 2021. Data access was provided by the District Hospital Review Board (DHRB) and the Department of Health (DOH), District Ambala, Haryana. Appropriate statistical tools were used to analyze the data. Statistical Package for Social Sciences (SPSS) 26.0 and Python 3.9 were used for statistical analysis and visualization, and a p-value of less than 0.05 was considered statistically significant. RESULTS: The total sample size of the study was 1,316, out of which 371 (28.2%) were vaccinated and 945 (71.8%) were not vaccinated. The mean age of the study participants was 49.6 ± 15.7 years. Seven hundred ninety-seven (60.6%) participants were male, while 519 (39.4%) participants were female. A statistically significant reduction was observed in the computed tomography severity score (CTSS) of the vaccinated population compared to the non-vaccinated group (χ2 = 74.3, p < 0.001). Vaccination led to a statistically significant decrease in mean CTSS across all lung lobes. CONCLUSION: Emerging COVID-19 variants challenge the effect of available vaccines, with different nations adopting different vaccination strategies to deal with the ongoing health problem. CTSS was employed as an objective marker to study the disease severity and effect of vaccination. Vaccination resulted in a significant reduction in CTSS seen on high-resolution computed tomography (HRCT) chest scans. There was a significant decrease in the incidence of severe COVID-19 pneumonia among vaccinated individuals. We need more observational data to corroborate the efficacy of vaccines presented in the randomized trials. Sharing such data between different nations can help us adopt a unifying vaccination strategy and decrease the impact of COVID-19 in subsequent disease waves.

Imbalanced IL-37/TNF-α/CTSS signaling disrupts corneal epithelial barrier in a dry eye model in vitro.

PURPOSE: To explore novel role and molecular mechanism of a natural anti-inflammatory cytokine interleukin (IL) 37 in preventing corneal epithelial barrier disruption from hyperosmolar stress as can occur in dry eye disease. METHODS: Primary human corneal epithelial cells (HCECs) were cultured from fresh donor limbal explants. An in vitro dry eye model with hyperosmolar stress was established by switching HCECs from isosmolar (312mOsM) to hyperosmolar medium (350-500 mOsM), and some cells were treated with rhIL-37 or rhTNF-α, for different periods (2-48 h). The expression of cytokines and cathepsin S, and barrier protein integrity were evaluated by RT-qPCR, ELISA, and immunofluorescent staining with confocal microscopy. RESULTS: The integrity of epithelial barrier was significantly disrupted in HCECs exposed to hyperosmolar medium, as shown by immunofluorescent images of tight junction (TJ, ZO-1, occludin and claudin-1) and adheren junction (E-cadherin) proteins. TNF-α accentuated hyperosmolar-induced disruption of TJ barrier functional integrity whereas exposure to IL-37 blunted or even reversed these changes. Cathepsin S, encoded by CTSS gene, was found to directly disrupt epithelial barrier integrity. Interestingly, CTSS expression was significantly induced by TNF-α and hyperosmolarity, while exogenous rhIL-37 inhibited TNF-α and CTSS expression at mRNA and protein levels following hyperosmolar stress. Furthermore, rhIL-37 restored barrier protein integrity, observed in 2D and 3D confocal immunofluorescent images, in HCECs under hyperosmolar stress. CONCLUSION: Our findings demonstrate a novel signaling pathway by which anti-inflammatory cytokine IL-37 prevents corneal epithelial barrier disruption under hyperosmotic stress via suppressing TNF-α and CTSS expression. This study provides new insight into mechanisms protecting corneal barrier in dry eye disease.

The RALE Score Versus the CT Severity Score in Invasively Ventilated COVID-19 Patients-A Retrospective Study Comparing Their Prognostic Capacities.

BACKGROUND: Quantitative radiological scores for the extent and severity of pulmonary infiltrates based on chest radiography (CXR) and computed tomography (CT) scan are increasingly used in critically ill invasively ventilated patients. This study aimed to determine and compare the prognostic capacity of the Radiographic Assessment of Lung Edema (RALE) score and the chest CT Severity Score (CTSS) in a cohort of invasively ventilated patients with acute respiratory distress syndrome (ARDS) due to COVID-19. METHODS: Two-center retrospective observational study, including consecutive invasively ventilated COVID-19 patients. Trained scorers calculated the RALE score of first available CXR and the CTSS of the first available CT scan. The primary outcome was ICU mortality; secondary outcomes were duration of ventilation in survivors, length of stay in ICU, and hospital-, 28-, and 90-day mortality. Prognostic accuracy for ICU death was expressed using odds ratios and Area Under the Receiver Operating Characteristic curves (AUROC). RESULTS: A total of 82 patients were enrolled. The median RALE score (22 [15-37] vs. 26 [20-39]; p = 0.34) and the median CTSS (18 [16-21] vs. 21 [18-23]; p = 0.022) were both lower in ICU survivors compared to ICU non-survivors, although only the difference in CTSS reached statistical significance. While no association was observed between ICU mortality and RALE score (OR 1.35 [95%CI 0.64-2.84]; p = 0.417; AUC 0.50 [0.44-0.56], this was noticed with the CTSS (OR, 2.31 [1.22-4.38]; p = 0.010) although with poor prognostic capacity (AUC 0.64 [0.57-0.69]). The correlation between the RALE score and CTSS was weak (r2 = 0.075; p = 0.012). CONCLUSIONS: Despite poor prognostic capacity, only CTSS was associated with ICU mortality in our cohort of COVID-19 patients.

Computed Tomography Severity Scoring on High-Resolution Computed Tomography Thorax and Inflammatory Markers With COVID-19 Related Mortality in a Designated COVID Hospital.

Introduction Radiological Society of the Netherlands introduced the coronavirus disease 2019 (COVID-19) Reporting and Data System (CO-RADS) and the corresponding CT severity score (CTSS) to diagnose COVID-19 severity. However, data regarding the same is very limited. Objectives The objective of this study was to correlate the computed tomography severity scoring (CTSS) on high-resolution computed tomography (HRCT) thorax and inflammatory markers with COVID-19 related mortality. Methods A retrospective observational study was conducted in a tertiary center between June 2020 to May 2021 among 2343 adult patients at the department of radio-diagnosis with suspected and confirmed COVID-19 cases who received an HRCT thorax. Data was collected retrospectively from the records regarding age, sex, and information regarding inflammatory markers such as C-reactive protein (CRP), ferritin, lactate dehydrogenase (LDH), erythrocyte sedimentation rate (ESR), D-dimer, and neutrophil-to-lymphocyte ratio. Information on HRCT thorax of patients was reviewed for radiological suspicion of COVID-19 related lung changes using CO-RADS scoring and severity of lung involvement using CT-severity scoring. Data was analyzed using SPSS version 22 (IBM Inc., Armonk, New York). Results The mean age was 51.69 ± 16.02 years, and most of the study participants were male (1592, 67.95%). The majority (999, 42.64%) had diabetes as a comorbidity. The reverse transcription polymerase chain reaction (RT-PCR) test was positive in 1571 (67.05%) participants. The majority (1571, 67.05%) had a CO-RADS score of six, and only 150 (6.40%) had CO-RADS score of four. The CT severity score was normal in 147 (6.27%), mild in 724 (30.90%), moderate in 903 (38.54%), and severe in 569 (24.29%) participants. The CRP levels were moderate in 1200 (51.22%) and severe in 428 (18.27%) participants. The mean ferritin, D-dimer and interleukin-6 (IL-6) levels were 321.83 ± 266.42 ng/ml, 1.51 ± 0.85mg/l, and 323.05 ± 95.52pg/ml, respectively. The mean length of hospital stay was 10.25 ± 6.52 days. Most of them (1926 out of 2343, 82.20%) survived, and nearly 417 out of 2343 (17.80%) died. Out of 2343, 569 participants had severe CT severity scores, out of which 205 (36.03%) died, and 364 (63.97%) participants survived. Conclusion A positive correlation was found between CT severity scoring on HRCT thorax and inflammatory markers with COVID-19 related mortality and can be used in early diagnosis and timely management of COVID-19 positive patients.

Spectrum of High Resolution Computed tomography chest findings in PCR positive COVID-19 patients according to duration of infection and CT severity score assessment.

OBJECTIVE: To evaluate the spectrum of HRCT findings of COVID-19 in RT-PCR positive patients according to duration of infection and severity of disease. METHODS: This retrospective study was conducted at Radiology department of Lahore General Hospital, Lahore from May to July 2020. Total 40 COVID-19 patients were reviewed for clinical features, HRCT chest findings based on time from symptom onset and CT conduction. Chi-square and fissure exact test were used for measuring association with severity of COVID-19, p value ≤0.05 was reported significant. Mean CT scores were calculated. ROC curve analysis showed threshold values of CT-SS for severe disease. RESULTS: Of total 40 patients with age ranged from 22-83 years, 22(55%) were males and 18(45%) females. The hallmark of COVID-19 was combined GGO and consolidation, GGO alone and consolidation alone in bilateral, sub pleural and posterior distribution. Early stage had normal CT or GGO alone, intermediate and late stage had both GGO and consolidation. Septal lines/bands and crazy paving pattern were prevalent in late stage. Clinically, 24 (60%) were in severe group and 16(40%) in mild group. Severity of COVID-19 was associated with GGO alone (p=0.05), GGO and consolidation (p=0.01), crazy paving (p=0.01) and lung scores (p≤0.05). The threshold values of CT-SS for identifying severe disease by two radiologists were 18.50 and 20.50. CONCLUSION: HRCT manifestations along with CT-SS aids in predicting disease severity. Staging according to duration of infection is effective in understanding variation in pattern of chest findings in coronavirus disease.

Value and prognostic impact of a deep learning segmentation model of COVID-19 lung lesions on low-dose chest CT.

Objectives: 1) To develop a deep learning (DL) pipeline allowing quantification of COVID-19 pulmonary lesions on low-dose computed tomography (LDCT). 2) To assess the prognostic value of DL-driven lesion quantification. Methods: This monocentric retrospective study included training and test datasets taken from 144 and 30 patients, respectively. The reference was the manual segmentation of 3 labels: normal lung, ground-glass opacity(GGO) and consolidation(Cons). Model performance was evaluated with technical metrics, disease volume and extent. Intra- and interobserver agreement were recorded. The prognostic value of DL-driven disease extent was assessed in 1621 distinct patients using C-statistics. The end point was a combined outcome defined as death, hospitalization>10 days, intensive care unit hospitalization or oxygen therapy. Results: The Dice coefficients for lesion (GGO+Cons) segmentations were 0.75±0.08, exceeding the values for human interobserver (0.70±0.08; 0.70±0.10) and intraobserver measures (0.72±0.09). DL-driven lesion quantification had a stronger correlation with the reference than inter- or intraobserver measures. After stepwise selection and adjustment for clinical characteristics, quantification significantly increased the prognostic accuracy of the model (0.82 vs. 0.90; p<0.0001). Conclusions: A DL-driven model can provide reproducible and accurate segmentation of COVID-19 lesions on LDCT. Automatic lesion quantification has independent prognostic value for the identification of high-risk patients.

Modulation of Cathepsin S (CTSS) Regulates the Secretion of Progesterone and Estradiol, Proliferation, and Apoptosis of Ovarian Granulosa Cells in Rabbits.

Cathepsin S (CTSS) is a member of cysteine protease family. Although many studies have demonstrated the vital role of CTSS in many physiological and pathological processes including tumor growth, angiogenesis and metastasis, the function of CTSS in the development of rabbit granulosa cells (GCS) remains unknown. To address this question, we isolated rabbit GCS and explored the regulatory function of the CTSS gene in cell proliferation and apoptosis. CTSS overexpression significantly promoted the secretion of progesterone (P4) and estrogen (E2) by increasing the expression of STAR and CYP19A1 (p < 0.05). We also found that overexpression of CTSS increased GCS proliferation by up-regulating the expression of proliferation related gene (PCNA) and anti-apoptotic gene (BCL2). Cell apoptosis was markedly decreased by CTSS activation (p < 0.05). In contrast, CTSS knockdown significantly decreased the secretion of P4 and E2 and the proliferation of rabbit GCS, while increasing the apoptosis of rabbit GCS. Taken together, our results highlight the important role of CTSS in regulating hormone secretion, cell proliferation, and apoptosis in rabbit GCS. These results might provide a basis for better understanding the molecular mechanism of rabbit reproduction.

Diagnostic Value of High-Resolution Computed Tomography Scan in COVID-19: Do We Need to Think Outside the Box?

Background and objective The ambiguous nature and high infectivity of the coronavirus disease 2019 (COVID-19) have caused soaring morbidity and mortality worldwide. Real-time polymerase chain reaction (RT-PCR) is preferred for detecting COVID-19. However, its poor sensitivity and the emerging use of high-resolution CT (HRCT) scan for disease severity make the use of RT-PCR quite obsolete. In light of this, our study aimed to explore the beneficial role of HRCT and compare the HRCT findings across various patient demographics and parameters. Methods This cross-sectional study included 100 patients with clinical suspicion of COVID-19. All patients underwent a chest HRCT scan preceded by RT-PCR testing. We used the CT severity score (CTSS) of the chest to calculate disease severity. Demographical data and results of radiological findings were tabulated and compared across RT-PCR positivity, age, and gender. Independent samples t-test and chi-square test were used to analyze the data. Results Glass ground opacity was the most prevalent finding in 99% of the patients, followed by lymph node involvement, consolidation, and crazy-paving pattern. Pleural effusion was observed in only 10% of the patients while pericardial effusion and hiatal hernia were present in 5%. In RT-PCR-positive patients, the posterior basal segment of the lower lobe of the right and left lungs were found to be dominantly involved; however, the upper and middle lobes of the right lung were more commonly involved than the left lung. The mean CTSS was significantly higher in patients aged above 50 years (p<0.001). The mean CTSS of RT-PCR-negative patients was higher than that of RT-PCR-positive patients (15.18 vs. 14.31, p=0.537). Conclusion RT-PCR has a limited role in the diagnosis of COVID-19. The HRCT scan can detect typical COVID-19 findings even in patients with negative RT-PCR results. Moreover, the use of HRCT scan in determining the disease severity and extent of lung damage can lead to a better assessment of critically ill patients.

Skullcapflavone II Suppresses TNF-α/IFN-γ-Induced TARC, MDC, and CTSS Production in HaCaT Cells.

Skullcapflavone II (SFII), a flavonoid derived from Scutellaria baicalensis, has been reported to have anti-inflammatory properties. However, its therapeutic potential for skin inflammatory diseases and its mechanism are unknown. Therefore, this study aimed to investigate the effect of SFII on TNF-α/IFN-γ-induced atopic dermatitis (AD)-associated cytokines, such as thymus- and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC). Co-stimulation with TNF-α/IFN-γ in HaCaT cells is a well-established model for induction of pro-inflammatory cytokines. We treated cells with SFII prior to TNF-α/IFN-γ-stimulation and confirmed that it significantly inhibited TARC and MDC expression at the mRNA and protein levels. Additionally, SFII also inhibited the expression of cathepsin S (CTSS), which is associated with itching in patients with AD. Using specific inhibitors, we demonstrated that STAT1, NF-κB, and p38 MAPK mediate TNF-α/IFN-γ-induced TARC and MDC, as well as CTSS expression. Finally, we confirmed that SFII significantly suppressed TNF-α/IFN-γ-induced phosphorylation of STAT1, NF-κB, and p38 MAPK. Taken together, our study indicates that SFII inhibits TNF-α/IFN-γ-induced TARC, MDC, and CTSS expression by regulating STAT1, NF-κB, and p38 MAPK signaling pathways.