The association of healthcare contact days with physical function and survival in CCTG/AGITG CO.17.

INTRODUCTION: While contact days-days with healthcare contact outside home-are increasingly adopted as a measure of time toxicity and treatment burden, they could also serve as a surrogate of treatment-related harm. We sought to assess the association between contact days and patient-reported outcomes, and the prognostic ability of contact days. METHODS: We conducted a secondary analysis of CO.17 that evaluated cetuximab vs supportive care in patients with advanced colorectal cancer. CO.17 collected EORTC-QLQ-C30 instrument data. We assessed the association between number of contact days in a window and changes in physical function and global health status, and the association between number of contact days in the first 4 weeks with overall survival (OS). RESULTS: There was a negative association between the number of contact days and change in physical function (per each additional contact day at 4 weeks, 1.50 point decrease; and 8 weeks, 1.06 point decrease, p < .0001 for both), but not with global health status. This negative association was seen in patients receiving cetuximab, but not supportive care. More contact days in the first 4 weeks was associated with worse OS for all comers and patients receiving cetuximab (per each additional contact day; all comers, aHR 1.07, 95% CI, 1.05- 1.10; and cetuximab, aHR 1.08, 95%CI 1.05- 1.11, p < .0001 for both). CONCLUSIONS: In this secondary analysis of a clinical trial, more contact days early in the course was associated with declines in physical function and worse survival in all-comers and in participants receiving cancer-directed treatment. TRIAL REGISTRATION: ClinicalTrials.gov number, NCT00079066.

Altered metabolome and microbiome associated with compromised intestinal barrier induced hepatic lipid metabolic disorder in mice after subacute and subchronic ozone exposure.

Exposure to ozone has been associated with metabolic disorders in humans, but the underlying mechanism remains unclear. In this study, the role of the gut-liver axis and the potential mechanism behind the metabolic disorder were investigated by histological examination, microbiome and metabolome approaches in mice during the subacute (4-week) and subchronic (12-week) exposure to 0.5 ppm and 2.5 ppm ozone. Ozone exposure resulted in slowed weight gain and reduced hepatic lipid contents in a dose-dependent manner. After exposure to ozone, the number of intestinal goblet cells decreased, while the number of tuft cells increased. Tight junction protein zonula occludens-1 (ZO-1) was significantly downregulated, and the apoptosis of epithelial cells increased with compensatory proliferation, indicating a compromised chemical and physical layer of the intestinal barrier. The hepatic and cecal metabolic profiles were altered, primarily related to lipid metabolism and oxidative stress. The abundance of Muribaculaceae increased dose-dependently in both colon and cecum, and was associated with the decrease of metabolites such as bile acids, betaine, and L-carnitine, which subsequently disrupted the intestinal barrier and lipid metabolism. Overall, this study found that subacute and subchronic exposure to ozone induced metabolic disorder via disturbing the gut-liver axis, especially the intestinal barrier. These findings provide new mechanistic understanding of the health risks associated with environmental ozone exposure and other oxidative stressors.

ctDNA response after pembrolizumab in non-small cell lung cancer: phase 2 adaptive trial results.

Circulating tumor DNA (ctDNA) has shown promise in capturing primary resistance to immunotherapy. BR.36 is a multi-center, randomized, ctDNA-directed, phase 2 trial of molecular response-adaptive immuno-chemotherapy for patients with lung cancer. In the first of two independent stages, 50 patients with advanced non-small cell lung cancer received pembrolizumab as standard of care. The primary objectives of stage 1 were to ascertain ctDNA response and determine optimal timing and concordance with radiologic Response Evaluation Criteria in Solid Tumors (RECIST) response. Secondary endpoints included the evaluation of time to ctDNA response and correlation with progression-free and overall survival. Maximal mutant allele fraction clearance at the third cycle of pembrolizumab signified molecular response (mR). The trial met its primary endpoint, with a sensitivity of ctDNA response for RECIST response of 82% (90% confidence interval (CI): 52-97%) and a specificity of 75% (90% CI: 56.5-88.5%). Median time to ctDNA response was 2.1 months (90% CI: 1.5-2.6), and patients with mR attained longer progression-free survival (5.03 months versus 2.6 months) and overall survival (not reached versus 7.23 months). These findings are incorporated into the ctDNA-driven interventional molecular response-adaptive second stage of the BR.36 trial in which patients at risk of progression are randomized to treatment intensification or continuation of therapy. ClinicalTrials.gov ID: NCT04093167 .

Antibiotic resistome and associated bacterial communities in agricultural soil following the amendments of swine manure-derived fermentation bed waste.

The practice of utilizing animal manures on land is widespread in agriculture, but it has raised concerns about the possible spread of antibiotic resistance genes (ARGs) and the potential risk it poses to public health through food production. Fermentation bed culture is an effective circular agricultural practice commonly utilized in pig farming that minimizes the environmental impact of livestock farming. However, this method generates a significant amount of fermentation bed waste (FBW), which can be turned into organic fertilizer for land application. The objective of this research was to examine the impacts of amending agricultural soil samples with swine manure-derived FBW on microbial communities, mobile genetic elements (MGEs), and ARG profiles over different periods. The study findings indicated that the amendment of swine manure-derived FBW significantly increased the diversity and abundance of ARGs and MGEs during the early stages of amendment, but this effect diminished over time, and after 12 months of FBW amendments, the levels returned to those comparable to control samples. The shift in the bacterial communities played a significant role in shaping the patterns of ARGs. Actinobacteriota, Proteobacteria, and Bacteroidetes were identified as the primary potential hosts of ARGs through metagenomic binning analysis. Furthermore, the pH of soil samples was identified as the most important property in driving the composition of the bacterial community and soil resistome. These findings provided valuable insights into the temporal patterns and dissemination risks of ARGs in FBW-amended agriculture soil, which could contribute to the development of effective strategies to manage the dissemination risks of FBW-derived ARGs.

Contact Days Associated With Cancer Treatments in the CCTG LY.12 Trial.

BACKGROUND: When cancer treatments have similar oncologic outcomes, the number of days with in-person healthcare contact (""contact days'') can help contextualize expected time use with each treatment. We assessed contact days in a completed randomized clinical trial. PATIENTS AND METHODS: We conducted a secondary analysis of the CCTG LY.12 RCT that evaluated 2-3 cycles of gemcitabine, dexamethasone, and cisplatin (GDP) vs. dexamethasone, cytarabine, and cisplatin (DHAP) in 619 patients with relapsed/refractory lymphoma prior to stem cell transplant. Primary analyses reported similar response rates and survival. We calculated patient-level "contact days" by analyzing trial forms. The study period was from assignment to progression or transplant. Days without healthcare contact were considered "home days''. We compared measures of contact days across arms. RESULTS: The study period was longer in the GDP arm (median 50, vs. 47 days, P = .007). Contact days were comparable in both arms (median 18 vs 19, P = 0.79), but home days were higher in the GDP arm (median 33 vs 28, P < .001). The proportion of contact days was lower in the GDP arm (34%, vs. 38%, P = .009). The GDP arm experienced more contact days related to planned outpatient chemotherapy (median, 10 vs. 8 days), but the DHAP arm experienced many more inpatient contact days (median, 11 vs. 0 days). CONCLUSIONS: Measures of time use, such as contact days, can be extracted from RCTs. In LY.12, despite comparable oncologic outcomes, GDP was associated with fewer contact days. Such information can guide decision-making for patients with hematological cancers, who already face significant healthcare contact.

Evaluating the Time Toxicity of Cancer Treatment in the CCTG CO.17 Trial.

PURPOSE: The time spent in pursuing treatments for advanced cancer can be substantial. We have previously proposed a pragmatic and patient-centered metric of these time costs-which we term time toxicity-as any day with physical health care system contact. This includes outpatient visits (eg, bloodwork, scans, etc), emergency department visits, and overnight stays in a health care facility. Herein, we sought to assess time toxicity in a completed randomized controlled trial (RCT). METHODS: We conducted a secondary analysis of the Canadian Cancer Trials Group CO.17 RCT that evaluated weekly cetuximab infusions versus supportive care alone in 572 patients with advanced colorectal cancer. Initial results reported a 6-week improvement in median overall survival (OS) with cetuximab (6.1 v 4.6 months). Subsequent analyses reported that benefit was restricted to patients with K-ras wild-type tumors. We calculated patient-level time toxicity by analyzing trial forms. We considered days without health care contact as home days. We compared medians of time measures across arms and stratified results by K-ras status. RESULTS: In the overall population, median time toxic days were higher in the cetuximab arm (28 v 10, P < .001) although median home days were not statistically different between arms (140 v 121, P = .09). In patients with K-ras-mutated tumors, cetuximab was associated with almost numerically equal home days (114 days v 112 days, P = .571) and higher time toxicity (23 days v 11 days, P < .001). In patients with K-ras wild-type tumors, cetuximab was associated with more home days (186 v 132, P < .001). CONCLUSION: This proof-of-concept feasibility study demonstrates that measures of time toxicity can be extracted through secondary analyses of RCTs. In CO.17, despite an overall OS benefit with cetuximab, home days were statistically similar across arms. Such data can supplement traditional survival end points in RCTs. Further work should refine and validate the measure prospectively.[Media: see text].

CD14, a novel surface marker of esophageal cancer stem cells.

Studies on targeting cancer stem cells (CSCs) have not yielded satisfactory results regarding solid tumor treatments; one of the reasons for this is the difficulty associated with the identification of a relatively specific antigen in solid tumors. CD14, which is mainly expressed in certain immune cells, is associated with tumor recurrence, growth, metastasis and resistance to treatment, which is in conformity with the characteristics of CSCs. It was thus hypothesized that esophageal CSCs (ECSCs) express CD14. In the present study, paraffin­embedded sections of human esophageal carcinoma were used to determine the co­expression of CD14 and the ECSC marker aldehyde dehydrogenase­1 (ALDH1) using immunofluorescence. CD14+ cells were then isolated using immunomagnetic separation for stemness detection, including proliferation, migration, invasion and tumorigenicity. Cell Counting Kit­8 (CCK­8), EdU and colony­formation assays were utilized to investigate the proliferative ability, the metastatic capacity was examined using Transwell and wound­healing assays and a xenograft assay was performed to investigate the tumorigenic ability. It was indicated that the ALDH1­labeled ECSCs expressed CD14 and primary CD14+ cells possessed the characteristics of CSCs. On the whole, the results of the present study suggest the potential utility of CD14 as a novel surface marker for ECSCs.

Diverse functional genes harboured in extracellular vesicles from environmental and human microbiota.

Exchange of mobile functional genes within microbiota benefits the microbial community. However, the status of the mobile gene pool in environment is still largely unclear, impeding the understanding on the process of gene transfer in natural microbial communities. The release of extracellular vesicles (EVs) by diverse organisms has been proposed to be a vital way in the complex networks of interactions between microbes and their habitats. In this study, we hypothesized that microbial EVs encapsulating functional DNA are widely distributed in the environmental matrix. The prevalence, source and DNA cargoes of EVs in three types of typical microbial habitats were studied. High abundance of EVs comparable to the bacterial concentration was found in human faeces, wastewater and soil. Metagenomic analysis showed the diverse and differential taxonomy of EVs-associated DNA compared to source microbiome. An array of efficient EVs producing species was identified. A wide variety of mobile genes including glycoside hydrolase family 25 were enriched. Antibiotic resistance genes co-localizing with mobile genetic elements were abundant in the EVs. This study provides novel insights into the prevalent EVs as a reservoir for the mobile functional genes in the natural environment.

Nephropathy 1st inhibits renal fibrosis by activating the PPARγ signaling pathway.

Renal fibrosis is a manifestation of kidney injury. Nephropathy 1st is a traditional Chinese herbal medicine that has been used as a therapy for kidney disease, but the underlying mechanisms remain elusive. The aim of this study was to investigate the role and underlying mechanisms of Nephropathy 1st on the progression of kidney disease. In the present study, unilateral ureteral obstruction was performed to establish the renal fibrosis rat model. By hematoxylin-eosin staining and immunohistochemical staining analysis, the severity of renal fibrosis was evaluated in vivo. Serum creatinine (CREA) and urea nitrogen (BUN) were measured by ELISA. The expression levels of Col-I, FN, PPARγ, and Klotho were measured by Western blot in rat NRK-49F cells and in fibrotic rats. GW9662 was used to inhibit PPARγ signaling. Metabonomic analysis showed metabolic differences among groups. Nephropathy 1st administration alleviated the progression of rat renal fibrosis and reduced serum creatinine (Scr) and BUN levels. Mechanistically, Nephropathy 1st promoted the expression of PPARγ and thus activated PPARγ signaling, thereby reducing the pro-fibrotic phenotypes of fibroblasts. The therapeutic effect of Nephropathy 1st was abrogated by the PPARγ inhibitor GW9662. Moreover, Nephropathy 1st normalized the dysregulated lipid metabolism in renal fibrosis rats. In conclusion, Nephropathy 1st alleviates renal fibrosis development in a PPARγ-dependent manner.

Correlation between lung cancer markers and air pollutants in western China population.

The relationship between serum lung cancer markers and the air pollution remains unclear. To further reveal the correlation between air pollutants and lung cancer, a retrospective analysis of 446,032 asymptomatic healthy people and symptomatic healthy people from the Health Management Center of the First Affiliated Hospital of Chongqing Medical University from 2014 to 2019 was performed. The distribution characteristics of serum lung cancer markers, cancer embryo antigens (CEA), cytokeratin 19 fragment (CYFRA211), squamous cell carcinoma antigen (SCC), and nerve-specific enolase (NSE) was analyzed in these population. Two independent sample man-Whitney U test was used to analyze the correlation of lung cancer markers and age, and a Chi-square test was used to analyze the relationship between lung cancer markers and gender. The daily change trend was profiled for six main air quality indicators PM10, PM2.5, SO2, NO2, CO, O3 during the same period. The correlation between lung markers and air pollutants was investigated by Spearman and multiple linear regression. The results showed that CYFRA211 had the highest excess rate in the screening population. There were differences in the number of cases with concentrated expression of lung cancer markers in the different age groups. Among them, the people with NSE exceeding the standard were the youngest, and most of them were 40-55 years old. Besides SCC, the expression levels of other markers increased with age, and the expression levels of the four markers in males were significantly higher than those in females. Although the levels of PM10 and PM2.5 exceeded the WHO standard (World Health Organization. 2011), they were not correlated with lung cancer markers. Multiple comparisons showed that the air pollutants SO2 and CYFRA211, as well as NO2 and NSE were closely related, but there was no significant linear relationship between CEA, SCC, and air pollutants. In conclusion, among the four lung cancer markers, CYFRA211 had the highest abnormal excess rate in total screening population, and the expression levels of these markers varied by gender and age, with males showing significantly higher expression levels than females, and they increased significantly with age except for SCC. The differential expression of these lung cancer markers may provide more strategies for lung cancer screening in the corresponding population. Lung cancer markers, CYFRA211 and NSE, can be used as sensitive biomarkers for exposure to certain air pollutants and provide references for the prevention and management of air pollution.

Widespread of Potential Pathogen-Derived Extracellular Vesicles Carrying Antibiotic Resistance Genes in Indoor Dust.

Extracellular vesicles (EVs) are newly recognized as important vectors for carrying and spreading antibiotic resistance genes (ARGs). However, the ARGs harbored by EVs in ambient environments and the transfer potential are still unclear. In this study, the prevalence of ARGs and mobile genetic elements (MGEs) in EVs and their microbial origins were studied in indoor dust from restaurants, kindergarten, dormitories, and vehicles. The amount of EVs ranged from 3.40 × 107 to 1.09 × 1011 particles/g dust. The length of EV-associated DNA fragments was between 21 bp and 9.7 kb. Metagenomic sequencing showed that a total of 241 antibiotic ARG subtypes encoding resistance to 16 common classes were detected in the EVs from all four fields. Multidrug, quinolone, and macrolide resistance genes were the dominant types. 15 ARG subtypes were exclusively carried and even enriched in EVs compared to the indoor microbiome. Moreover, several ARGs showed co-occurrence with MGEs. The EVs showed distinct taxonomic composition with their original dust microbiota. 30.23% of EV-associated DNA was predicted to originate from potential pathogens. Our results indicated the widespread of EVs carrying ARGs and virulence genes in daily life indoor dust, provided new insights into the status of extracellular DNA, and raised risk concerns on their gene transfer potential.

Dataset-based assessment of heavy metal contamination in freshwater fishes and their health risks.

The ecological risks and health hazards of heavy metals pollution in Taihu Lake have received widespread concern. This study has developed a species-pollution dataset which includes a large amount of data on heavy metal pollution in Taihu fish. The heavy metal contamination poses a significant threat to human consumption, but no studies have been conducted to assess the risk of exposure to consumption of these fish and to make recommendations for their consumption. In this study, we systematically integrated the relevant data in the dataset, analyzed its contamination level using PI (single pollution index) and MPI (metal pollution index) models, and assessed health hazards of fish consumption using THQ (target hazard quotient) and ILCR (incremental lifetime cancer risk) models. Results showed that the contamination levels of heavy metals in fish varied in a feeding habit and living habit dependent manner. The risk of non-cancer health is the highest from consuming omnivorous fish, then from carnivorous and herbivorous fish. The ILCR model predicted that the long-term Taihu consumption of omnivorous fish may pose a potential carcinogenic risk, especially for children. In all, our study provided a comprehensive understanding on the risk of heavy metals in Taihu. Accordingly, it is recommended that children should try to choose herbivorous fish when consuming fish from Taihu Lake while avoiding long-term consumption of omnivorous fish.

Expression and immunogenicity of nsp10 protein of porcine epidemic diarrhea virus.

Porcine epidemic diarrhea virus (PEDV), a swine enteropathogenic coronavirus, causes lethal watery diarrhea to the piglets, which poses significant economic losses and public health concerns. The nsp10 protein of PEDV is essential regulatory subunits that are critical for virus replication. Since PEDV nsp10 is a crucial regulator of viral RNA synthesis, it is promising that nsp10 might become anti-virus drugs target or candidate for rapid diagnosis of PEDV infection. In this study, the PEDV nsp10 was inserted into pMAL-c2x-MBP / pET-28a vector, efficiently and stably expressed in E.coli system. Then the purified nsp10 protein was found to mediate potent antibody responses in immunized mice. The antibodies of immunized mice and PEDV infection swine strongly recognized purified nsp10 protein from cell lysates. Furthermore, cytokines test revealed that the expression of IL-2, IL-4, IL-10, TNF-α, IFN-γ were significantly higher than those in control group, indicated that purified nsp10 protein induce the cellular immune response mechanism in mice. Using modified seroneutralization test, we also demonstrated that sera from nsp10-immunized mice inhibited PEDV replication to some extent. These findings suggest that nsp10 has a high immunogenicity. This study may have implications for future development of PEDV detection or anti-virus drugs for swine.

The impact of air pollution on COVID-19 pandemic varied within different cities in South America using different models.

There is a rising concern that air pollution plays an important role in the COVID-19 pandemic. However, the results were not consistent on the association between air pollution and the spread of COVID-19. In the study, air pollution data and the confirmed cases of COVID-19 were both gathered from five severe cities across three countries in South America. Daily real-time population regeneration (Rt) was calculated to assess the spread of COVID-19. Two frequently used models, generalized additive models (GAM) and multiple linear regression, were both used to explore the impact of environmental pollutants on the epidemic. Wide ranges of all six air pollutants were detected across the five cities. Spearman's correlation analysis confirmed the positive correlation within six pollutants. Rt value showed a gradual decline in all the five cities. Further analysis showed that the association between air pollution and COVID-19 varied across five cities. According to our research results, even for the same region, varied models gave inconsistent results. For example, in Sao Paulo, both models show SO2 and O3 are significant independent variables, however, the GAM model shows that PM10 has a nonlinear negative correlation with Rt, while PM10 has no significant correlation in the multiple linear model. Moreover, in the case of multiple regions, currently used models should be selected according to local conditions. Our results indicate that there is a significant relationship between air pollution and COVID-19 infection, which will help states, health practitioners, and policy makers in combating the COVID-19 pandemic in South America.

Oxidative degradation/mineralization of dimethyl phthalate (DMP) from plastic industrial wastewater using ferrate(VI)/TiO2 under ultraviolet irradiation.

A novel ferrate(VI)/titanium dioxide/ultraviolet [Fe(VI)/TiO2/UV] system was successfully established for the photocatalytic oxidation of dimethyl phthalate (DMP). This system demonstrated a higher removal efficiency of DMP (95.2%) than the conventional TiO2/UV and Fe(VI) alone systems (51.8% and 23.5%, respectively) and produced obvious synergistic effects. Response surface methodology (RSM), based on a three level, three independent variables design, was conducted through Design Expert 8.0.6 program, and a second-order polynomial model (R2 = 0.998) was developed to quantitatively describe the photocatalysis of TiO2 combined with Fe(VI) oxidation under ultraviolet irradiation. The fresh TiO2 and photochemical reacted Fe(VI)/TiO2 were characterized by X-ray diffraction (XRD), scanning electron microscope (SEM), and element dispersive spectrum (EDS), which indicated that Fe(VI) was imprinted into the TiO2, and the surface adsorbed Fe-O-(organic) materials inhibited DMP degradation. This photocatalytic oxidant showed high activity and stability after nine cycles without loss of its effectiveness (counting from the second cycle). The intermediates/products of DMP were analyzed by gas chromatography-mass spectrometry. The proposed pathway for DMP degradation involved one electron transfer of hydroxyl radical and breaking of the ester bond and benzene ring. The mineralization efficiencies of DMP in actual industrial wastewater and simulated water were 87.1% and 95.2%, respectively, suggesting practical field applications. A ecotoxicity test (17.3% inhibition on bioluminescence) in treating actual industrial wastewater containing DMP implied that the proposed Fe(VI)/TiO2/UV had a potential for industrial water treatment.

Anlotinib Suppresses Oral Squamous Cell Carcinoma Growth and Metastasis by Targeting the RAS Protein to Inhibit the PI3K/Akt Signalling Pathway.

Oral squamous cell carcinoma (OSCC) is a malignant tumour originating from the mucosal lining of the oral cavity. Its characteristics include hidden onset, high recurrence, and distant metastasis after operation. At present, clinical treatment usually includes surgery, chemotherapy, radiotherapy, or the joint use of these modalities. Unfortunately, multidrug resistant is one of the important obstacles that causes cancer chemotherapy failure. Anlotinib, which has recently been proven to have good antitumour effects, is a novel multitargeted tyrosine kinase inhibitor. However, there are few studies of the anlotinib-associated mechanism in OSCC and its underlying molecular mechanism. In our study, in vitro models of human oral squamous cell carcinoma HSC-3 cells were used to determine the efficacy of anlotinib. On the one hand, we showed that anlotinib treatment significantly reduced the viability and proliferation of HSC-3 cells and decreased cell migration by inhibiting the activation of the Akt phosphorylation pathway. On the other side, anlotinib inhibited PI3K/Akt/Bad phosphorylation and promoted apoptosis of HSC-3 cells by activating RAS protein expression. In brief, these results indicated that anlotinib had prominent antitumour activity in OSCC, mainly by inhibiting the PI3K/Akt phosphorylation pathway. This work provides evidences and a basic principle for using anlotinib to treat patients with OSCC for clinical research.

Acid resistance of Masson pine (Pinus massoniana Lamb.) families and their root morphology and physiological response to simulated acid deposition.

Pinus massoniana Lamb. is one of the most sensitive species to acid deposition among forest woody plants, but differences in acid resistance among pine families still exist. It is of great significance to study the differences in acid resistance of Masson pine families and to analyze the physiological regulation mechanism of their acid resistance. In this study, the 100-day-old seedlings of 16 Masson pine families were treated with the simulated acid rain (SAR) at different pH levels (5.6, 4.5, 3.5 and 2.5) for 100 days to investigate the plant morphology, chlorophyll content, and root physiological responses. Results showed that pine family No. 35 maintained the good morphology, high chlorophyll content and organic acids secretion, and low plasma membrane permeability exposed to SAR, while family No. 79 presented the opposite. SAR not only increased the root plasma membrane permeability, but also induced an exudation of organic acids from the pine roots, and the test parameters changed sharply when the SAR pH was lower than 4.5. The results indicated that Masson pine could resist to acidic environment (pH 4.5-5.6), and family No. 35 had the acid resistance while the family No. 79 was sensitive to acid stress. The acid resistance diversity of different pine families had close relation with the root physiological processes, including the root plasma membrane permeability and organic acids secretion. For the future research, the natural genetic variation of Masson pine in response to acid stress and its acid resistance mechanism should be further studied.

Higher serum PD-L1 level predicts increased overall survival with lapatinib versus trastuzumab in the CCTG MA.31 phase 3 trial.

BACKGROUND: The purpose of this retrospective biomarker study of the Canadian Cancer Trials Group (CCTG) MA.31 randomized phase 3 trial (lapatinib vs trastuzumab) of HER2-positive metastatic breast cancer (MBC) was to evaluate the prognostic and predictive biomarker utility of pretreatment serum programmed death ligand 1 (PD-L1) levels. METHODS: CCTG MA.31 accrued 652 HER2-positive patients; 387 had serum available (185 in the trastuzumab arm and 202 in the lapatinib arm). The Ella immunoassay platform (ProteinSimple, San Jose, California) was used to quantitate serum PD-L1 levels. Stepwise forward Cox multivariable analyses were performed for progression-free survival and overall survival (OS). RESULTS: In the whole trial population, continuous pretreatment serum PD-L1 levels were not associated with OS. However, within the trastuzumab arm, a higher continuous pretreatment serum PD-L1 level was significant for shorter OS (hazard ratio [HR], 3.85; P = .04), but within the lapatinib arm, pretreatment serum PD-L1 was not associated with OS (P = .37). In the whole trial, in a multivariable analysis for OS, serum PD-L1 (median cut point) remained a significant independent covariate (HR, 2.38; P = .001). There was a significant interaction between treatment arm and continuous serum PD-L1 (bootstrap method; P = .0025): at or above 214.2 pg/mL (the 89th percentile), serum PD-L1 was associated with significantly shorter OS with trastuzumab treatment versus lapatinib treatment. CONCLUSIONS: In the CCTG MA.31 trial, serum PD-L1 was a significant predictive factor: a higher pretreatment serum PD-L1 level was associated with shorter OS with trastuzumab treatment but with longer OS with lapatinib treatment. Immune evasion may decrease the effectiveness of trastuzumab therapy. Further evaluation of elevated serum PD-L1 in advanced breast cancer is warranted to identify patients with HER2-positive MBC who may benefit from novel immune-targeted therapies in addition to trastuzumab.

Meteorological impact on the COVID-19 pandemic: A study across eight severely affected regions in South America.

The role of meteorological factors in the transmission of the COVID-19 still needs to be determined. In this study, the daily new cases of the eight severely affected regions in four countries of South America and their corresponding meteorological data (average temperature, maximum temperature, minimum temperature, average wind speed, visibility, absolute humidity) were collected. Daily number of confirmed and incubative cases, as well as time-dependent reproductive number (Rt) was calculated to indicate the transmission of the diseases in the population. Spearman's correlation coefficients were assessed to show the correlation between meteorological factors and daily confirmed cases, daily incubative cases, as well as Rt. In particular, the results showed that there was a highly significant correlation between daily incubative cases and absolute humidity throughout the selected regions. Multiple linear regression model further confirmed the negative correlation between absolute humidity and incubative cases. The absolute humidity is predicted to show a decreasing trend in the coming months from the meteorological data of recent three years. Our results suggest the necessity of continuous controlling policy in these areas and some other complementary strategies to mitigate the contagious rate of the COVID-19.

Japanese Encephalitis Virus Induces Apoptosis and Encephalitis by Activating the PERK Pathway.

Accumulated evidence demonstrates that Japanese encephalitis virus (JEV) infection triggers endoplasmic reticulum (ER) stress and neuron apoptosis. ER stress sensor protein kinase R-like endoplasmic reticulum kinase (PERK) has been reported to induce apoptosis under acute or prolonged ER stress. However, the precise role of PERK in JEV-induced apoptosis and encephalitis remains unknown. Here, we report that JEV infection activates the PERK-ATF4-CHOP apoptosis pathway both in vitro and in vivo PERK activation also promotes the formation of stress granule, which in turn represses JEV-induced apoptosis. However, PERK inhibitor reduces apoptosis, indicating that JEV-activated PERK predominantly induces apoptosis via the PERK-ATF4-CHOP apoptosis pathway. Among JEV proteins that have been reported to induce ER stress, only JEV NS4B can induce PERK activation. PERK has been reported to form an active molecule by dimerization. The coimmunoprecipitation assay shows that NS4B interacts with PERK. Moreover, glycerol gradient centrifugation shows that NS4B induces PERK dimerization. Both the LIG-FHA and the LIG-WD40 domains within NS4B are required to induce PERK dimerization, suggesting that JEV NS4B pulls two PERK molecules together by simultaneously interacting with them via different motifs. PERK deactivation reduces brain cell damage and encephalitis during JEV infection. Furthermore, expression of JEV NS4B is sufficient to induce encephalitis via PERK in mice, indicating that JEV activates PERK primarily via its NS4B to cause encephalitis. Taken together, our findings provide a novel insight into JEV-caused encephalitis.IMPORTANCE Japanese encephalitis virus (JEV) infection triggers endoplasmic reticulum (ER) stress and neuron apoptosis. ER stress sensor protein kinase R-like endoplasmic reticulum kinase (PERK) has been reported to induce apoptosis under acute or prolonged ER stress. However, whether the PERK pathway of ER stress response plays important roles in JEV-induced apoptosis and encephalitis remains unknown. Here, we found that JEV infection activates ER stress sensor PERK in neuronal cells and mouse brains. PERK activation induces apoptosis via the PERK-ATF4-CHOP apoptosis pathway upon JEV infection. Among the JEV proteins prM, E, NS1, NS2A, NS2B, and NS4B, only NS4B activates PERK. Moreover, activated PERK participates in apoptosis and encephalitis induced by JEV and NS4B. These findings provide a novel therapeutic approach for JEV-caused encephalitis.