How to investigate human health effects related to exposure to mixtures of per- and polyfluoroalkyl substances: A systematic review of statistical methods.

BACKGROUND: Humans are exposed to several per- and polyfluoroalkyl substances (PFAS) daily; however, most previous studies have focused on individual PFAS. Although attention to effects of exposure to mixtures of PFAS has grown in recent years, there is no consensus on the appropriate statistical methods that can be used to assess their combined effect on human health. OBJECTIVES: We aim to perform a comprehensive review of the statistical methods used in the existing studies which evaluate the association between exposure to mixtures of PFAS and any adverse human health effect. METHODS: The online databases PubMed, Embase and Scopus were searched for eligible studies, published during the last ten years (last search performed on April 08, 2021). Covidence software was used by two different reviewers to perform a title/abstract screening, followed by a full text revision of the selected papers. RESULTS: A total of 3640 papers were identified, and after the screening process, 53 papers were included in the current review. Most of the studies were published between 2019 and 2021 and were conducted mainly in North America and Europe; more than half of the studies (28 out of 53) were conducted on mother and child pairs. WQS (Weighted Quantile Sum) Regression and BKMR (Bayesian Kernel Machine Regression) were used in 36 out of 53 papers to model mixtures' effects. Health outcomes included in the studies are immunotoxicity (n = 8), fetal development (n = 7), neurodevelopment (n = 9), reproductive hormones (n = 6), thyroid hormones (n = 7), outcomes related to metabolic pathways (n = 16). CONCLUSION: Studies on human exposure to PFAS as complex mixtures and health consequences have substantially increased in the last few years. Based on our findings, we propose that addressing risk from PFAS mixtures will likely require combinations of approaches and implementation of constantly evolving statistical methods. Specific guidelines and tools for quality assessment and publication of mixture observational studies are warranted.

Ginkgolic Acid Inhibits Coronavirus Strain 229E Infection of Human Epithelial Lung Cells.

Since December 2019, the COVID-19 pandemic has affected more than 200 million individuals around the globe and caused millions of deaths. Although there are now multiple vaccines for SARS-CoV-2, their efficacy may be limited by current and future viral mutations. Therefore, effective antiviral compounds are an essential component to win the battle against the family of coronaviruses. Ginkgolic Acid (GA) is a pan-antiviral molecule with proven effective in vitro and in vivo activity. We previously demonstrated that GA inhibits Herpes Simplex Virus 1 (HSV-1) by disrupting viral structure, blocking fusion, and inhibiting viral protein synthesis. Additionally, we reported that GA displays broad-spectrum fusion inhibition encompassing all three classes of fusion proteins, including those of HIV, Ebola, influenza A, and Epstein Barr virus. Here, we report that GA exhibited potent antiviral activity against Human Coronavirus strain 229E (HCoV-229E) infection of human epithelial lung cells (MRC-5). GA significantly reduced progeny virus production, expression of viral proteins, and cytopathic effects (CPE). Furthermore, GA significantly inhibited HCoV-229E even when added post-infection. In light of our findings and the similarities of this family of viruses, GA holds promising potential as an effective antiviral treatment for SARS-CoV-2.

Multifaceted Role of AMPK in Viral Infections.

Viral pathogens often exploit host cell regulatory and signaling pathways to ensure an optimal environment for growth and survival. Several studies have suggested that 5'-adenosine monophosphate-activated protein kinase (AMPK), an intracellular serine/threonine kinase, plays a significant role in the modulation of infection. Traditionally, AMPK is a key energy regulator of cell growth and proliferation, host autophagy, stress responses, metabolic reprogramming, mitochondrial homeostasis, fatty acid ß-oxidation and host immune function. In this review, we highlight the modulation of host AMPK by various viruses under physiological conditions. These intracellular pathogens trigger metabolic changes altering AMPK signaling activity that then facilitates or inhibits viral replication. Considering the COVID-19 pandemic, understanding the regulation of AMPK signaling following infection can shed light on the development of more effective therapeutic strategies against viral infectious diseases.

Excess of all-cause mortality is only partially explained by COVID-19 in Veneto (Italy) during spring outbreak.

BACKGROUND: Italy has been the first European country to be affected by the COVID-19 epidemic which started out at the end of February. In this report, we focus our attention on the Veneto Region, in the North-East of Italy, which is one of the areas that were first affected by the rapid spread of SARS-CoV-2. We aim to evaluate the trend of all-cause mortality and to give a description of the characteristics of the studied population. METHODS: Data used in the analyses were released by the majority of municipalities and cover the 93% of the total population living in the Veneto Region. We evaluated the trend of overall mortality from Jan.01 to Jun.30. 2020. Moreover we compared the COVID-19-related deaths to the overall deaths. RESULTS: From March 2020, the overall mortality rate increased exponentially, affecting males and people aged > 76 the most. The confirmed COVID-19-related death rate in the Veneto region between Mar.01 and Apr.302020 is 30 per 100,000 inhabitants. In contrast, the all-cause mortality increase registered in the same months in the municipalities included in the study is 219 per 100,000 inhabitants. CONCLUSIONS: COVID-19 has a primary role in the increase in mortality but does not entirely explain such a high number of deaths. Strategies need to be developed to reduce this gap in case of future waves of the pandemic.

[Gender and age factors affecting the mortality during the COVID-19 epidemic in Italy]. / Effetti del genere e dell'età sugli eccessi di mortalità durante l'epidemia di COVID-19 in Italia.

BACKGROUND: the COVID-19 epidemic severely affected Italy among European countries causing a considerable number of deaths across the country, especially in Northern Italy, leading also to serious problems to the Italian healthcare system, in particular the overcrowding of Intensive Care Units (ICU). In literature, the debate on the overall mortality during the COVID-19 epidemic, directly and indirectly, associated with the disease, is still open. OBJECTIVES: to describe the time trend of the mortality in Italy during the COVID-19 pandemic accounting for age, gender, and geographical areas. DESIGN: analysis of mortality trend, by region, age, and gender. SETTING AND PARTICIPANTS: the Italian mortality data, released by the Italian National Institute of Statistics (Istat), have been considered for the analyses. The data refer to the first four months of 2015-2019 and 2020, involving 7,270 municipalities, corresponding to 93% of the Italian population. MAIN OUTCOME MEASURES: the mortality rates in the first four months of 2015-2019 and 2020, age-adjusted, have been calculated together with the percent variation. The data were analysed by gender, age class (<65; >=65 years), Region, and geographical areas (Northern versus Central-Southern Italy). The overall daily mortality series have been represented as rates over 100,000 resident population. RESULTS: in addition to the geographical location, the age component was a major determinant of the mortality pattern. The greater increase in the overall mortality was evidenced among elderly subjects in the Northern Italian Regions most affected by the epidemic. In these areas, also gender component played an important role in determining the mortality excess: higher mortality rates in the first four months of 2020 are observed for males in comparison to female populations. CONCLUSIONS: this research reveals that the population components are an important issue in determining the COVID-19 mortality excess. For this reason, it is of primary importance to monitor mortality (overall and by COVID-19) by age and gender and to consider these components and the related factors (comorbidity, exposures affecting the lung) in the public prevention policies towards the protection of the most fragile population groups.

Ginkgolic acid inhibits fusion of enveloped viruses.

Ginkgolic acids (GA) are alkylphenol constituents of the leaves and fruits of Ginkgo biloba. GA has shown pleiotropic effects in vitro, including: antitumor effects through inhibition of lipogenesis; decreased expression of invasion associated proteins through AMPK activation; and potential rescue of amyloid-ß (Aß) induced synaptic impairment. GA was also reported to have activity against Escherichia coli and Staphylococcus aureus. Several mechanisms for this activity have been suggested including: SUMOylation inhibition; blocking formation of the E1-SUMO intermediate; inhibition of fatty acid synthase; non-specific SIRT inhibition; and activation of protein phosphatase type-2C. Here we report that GA inhibits Herpes simplex virus type 1 (HSV-1) by inhibition of both fusion and viral protein synthesis. Additionally, we report that GA inhibits human cytomegalovirus (HCMV) genome replication and Zika virus (ZIKV) infection of normal human astrocytes (NHA). We show a broad spectrum of fusion inhibition by GA of all three classes of fusion proteins including HIV, Ebola virus (EBOV), influenza A virus (IAV) and Epstein Barr virus (EBV). In addition, we show inhibition of a non-enveloped adenovirus. Our experiments suggest that GA inhibits virion entry by blocking the initial fusion event. Data showing inhibition of HSV-1 and CMV replication, when GA is administered post-infection, suggest a possible secondary mechanism targeting protein and DNA synthesis. Thus, in light of the strong effect of GA on viral infection, even after the infection begins, it may potentially be used to treat acute infections (e.g. Coronavirus, EBOV, ZIKV, IAV and measles), and also topically for the successful treatment of active lesions (e.g. HSV-1, HSV-2 and varicella-zoster virus (VZV)).