Systemic Medications of Dermatological Importance in COVID-19

Early December 2019 witnessed an international outbreak of a novel coronavirus (COVID 19) designated severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2). Since then, a number of therapeutic molecules have been explored to have potential efficacy against the SARS-Cov-2 per se or its sequelae. There are no Food and Drug Administration specific therapies approved so far;however, numerous drugs based on varying levels of evidence, in vitro studies and compassionate drug trials are being established as therapeutic agents, especially drugs approved for previous emergence of the severe acute respiratory syndrome (SARS-CoV-1) and Middle east respiratory syndrome coronavirus (MERS-Cov). Numerous active clinical trials for COVID-19 with more than 150 drugs and products are under study. Needless to say, many dermatological drugs are being employed to mitigate this pandemic threat. We aim to review drugs with potential against SARS-Cov-2 widely used in dermatology practice. Additionally, rampant and overzealous use of these drugs as well as introduction of new molecules might lead to emergence of adverse effects associated with these agents. Dermatologists must be on lookout for any cutaneous adverse effects of these drugs.Copyright © 2020.

NICARAGUA: Populist Performance and Authoritarian Practice During COVID-19

COVID-19 has ravaged Latin America’s poorest country, Nicaragua, which suffered among the highest excess mortality rates in the world at the pandemic’s height in mid-2020. While it would be easy to blame underdevelopment, weak public health infrastructure, and the lack of healthcare access for the pandemic’s toll, the authoritarian nature of the ruling regime, led by President Daniel Ortega and his wife and Vice President Rosario Murillo, exacerbated the devastation. Having centralized power through brutal repression since 2018, the Ortega-Murillo regime’s strategy was characterized by denial of the pandemic’s severity, the distortion of information, and the criminalization of the medical community and citizen-led responses efforts. Beyond authoritarianism, the regime’s approach demonstrated the key features of populist crisis performance: (1) the invocation of “the people” as a means of repressing opposition and rejecting a stronger pandemic response as economically disastrous and (2) the perpetuation of the crisis to further consolidate regime power. While downplaying the pandemic and resisting preventive measures were largely driven by economic considerations, increased repression amid waning support reflected the regime’s attempts to eliminate any alternative sources of popular legitimacy and preserve the ruling family’s grip on power and wealth. © 2023 selection and editorial matter, Nils Ringe and Lucio Rennó;individual chapters, the contributors.

Inhibition of estrogen receptor signaling as a strategy for radiosensitization of ER+ breast cancers

Purpose: The estrogen receptor (ER) is expressed in over 80% of breast tumors and has been shown to be a significant driver of breast cancer (BC) pathogenesis and therefore a target of effective first-line therapies. While both ionizing radiation (RT) and endocrine therapies (ET) are used for the treatment of ER+ BC, the effect of ET on tumor radiosensitization remains unclear, with concerns it may be radioprotective based on G1 cell arrest with ET treatment. Here we assessed the efficacy and mechanism of ER-mediated radiosensitization using various pharmacologic approaches in ER+ BC. Methods: Radiosensitization with ER inhibitors (tamoxifen [TAM], fulvestrant [FULV], AZD9496) was assessed using clonogenic survival assays. DNA damage was assessed by the neutral comet assay. Efficiency of homologous recombination (HR) or non-homologous end joining (NHEJ) as well as changes in cell cycle, apoptosis, and senescence were assessed. The efficacy of TAM with RT in vivo was assessed with an MCF-7 xenograft model. Results: The selective estrogen receptor modulator TAM radiosensitized ER+ MCF-7 (enhancement ratio [enhR]: 1.14-1.50) and T47D (enhR: 1.33-1.60) cells but not ER-negative SUM-159 cells (enhR: 0.99-1.02). The selective estrogen receptor degrader (SERD) FULV had similar radiosensitization effects in MCF-7 (enhR: 1.33-1.76) and T47D cells (enhR: 0.97-2.81) with no radiosensitization observed in SUM-159 cells (enhR: 1.01-1.03). The novel oral SERD AZD9496 radiosensitized MCF-7 cells (enhR: 1.36-1.56). MCF-7 cells treated with TAM and RT had an increase in dsDNA breaks compared to RT alone as measured by the comet assay (p<0.05) and a decrease in NHEJ-mediated repair with TAM (p<0.05). No changes were observed in HR-mediated repair by Rad51 foci or a reporter (p=NS). RT alone and in combination with TAM or FULV induced similar levels of cell cycle arrest, suggesting that radiosensitization with the combination therapy is cell-cycle independent. There were no significant changes in apoptosis with TAM, FULV, RT, or the combination (p=NS). Although TAM or FULV did induce senescence, ET with RT increased senescence induction (p<0.05). In vivo, combination RT and TAM led to a significant delay in days to tumor doubling (control: 17, TAM: 40, RT: 32, TAM+RT: undefined;p<0.0001), and a significant difference in tumor growth between mice treated with TAM or RT alone compared combination treatment, with no increased toxicities or skin lesions from the combination treatment. Conclusion: Our data suggest that TAM, FULV, or AZD9496 can radiosensitize ER+ breast tumors, and these agents with RT may be more effective for radiosensitization. This work also supports further clinical investigation of the timing of RT for patients receiving ET, including using ET during RT, especially as initiating ET prior to RT has been increasingly utilized as a bridging therapy followed by concurrent ET+RT during the COVID-19 pandemic.

GUATEMALA 2021: RECONSOLIDATING IMPUNITY AND REVERSING DEMOCRACY

The year 2021 marked the 25th anniversary of the signing of Guatemala's landmark peace accords. Yet celebrations of this historic achievement stood in sharp contrast to the year's alarming trends in the deterioration of democracy and the rule of law. Reviewing the events of 2021, this article uncovers two key patterns in Guatemalan politics, both evidence of democratic decline: 1) open assaults on judicial independence and breaches of the constitutional order, and 2) an escalation in the criminalization of government critics, particularly those on the frontlines of the anti-corruption struggle. It also takes stock of the most pernicious consequences of these patterns, including the abysmal Covid-19 response and the new opportunities for corruption amid resurgent impunity. Combined, these dynamics have also increased tensions within US-Guatemala relations as the Biden White House increasingly perceived the Guatemalan government as an unreliable partner in tackling the root causes of migration. In sum, 2021 was an inflection point in Guatemala's 25-year trajectory of peace-one that has steadily sent the country down an authoritarian path like many of its neighbors in the Western Hemisphere.

Are intravitreal injections essential during the COVID-19 pandemic? Global preferred practice patterns and practical recommendations.

Tertiary outpatient ophthalmology clinics are high-risk environments for COVID-19 transmission, especially retina clinics, where regular follow-up is needed for elderly patients with multiple comorbidities. Intravitreal injection therapy (IVT) for chronic macular diseases, is one of the most common procedures performed, associated with a significant burden of care because of the vigorous treatment regimen associated with multiple investigations. While minimizing the risk of COVID-19 infection transmission is a priority, this must be balanced against the continued provision of sight-saving ophthalmic care to patients at risk of permanent vision loss. This review aims to give evidence-based guidelines on managing IVT during the COVID-19 pandemic in common macular diseases such as age-related macular degeneration, diabetic macula edema and retinal vascular disease and to report on how the COVID-19 pandemic has affected IVT practices worldwide.To illustrate some real-world examples, 18 participants in the International Retina Collaborative, from 15 countries and across four continents, were surveyed regarding pre- and during- COVID-19 pandemic IVT practices in tertiary ophthalmic centers. The majority of centers reported a reduction in the number of appointments to reduce the risk of the spread of COVID-19 with varying changes to their IVT regimen to treat various macula diseases. Due to the constantly evolving nature of the COVID-19 pandemic, and the uncertainty about the normal resumption of health services, we suggest that new solutions for eye healthcare provision, like telemedicine, may be adopted in the future when we consider new long-term adaptations required to cope with the COVID-19 pandemic.

DETECTION OF SARS-COV-2 IN STOOL BUT NOT NARES OF NEWBORNS OF MOTHERS WITH COVID-19 DURING PREGNANCY

Purpose of Study To investigate potential evidence of in utero transmission of SARS-CoV-2 in the stool of newborns born to mothers with COVID-19 infection during pregnancy. Methods Used We investigated stool from 1 day to 2 months of age from 14 newborns born at 25-41 weeks whose mothers had COVID-19 during pregnancy. Newborns were admitted at delivery to the NICU or newborn nursery of our urban academic hospital from July 2020 to May 2021. A comparison group of 30 newborns had similar GAs and were born to mothers without COVID-19 during pregnancy. SARS-CoV-2 RNA was quantified with quantitative PCR using primers against SARS-CoV-2 envelope protein and non-structural protein 14 (NSP-14), spike protein with enzyme-linked immunosorbent assay (ELISA), and inflammatory cytokines interleukin- 6 (IL-6) and interferon-g (IFN-γ) elicited by stool homogenates in mouse bone marrow macrophages. This study was IRB approved with parental consent. Summary of Results Despite negative nasal PCRs from all newborns, viral RNAs and spike protein were detected in the stool of 11 out of 14 newborns as early as the first day of life (range 0-2 months, figure 2A and 2B). Stool RNA and spike protein levels increased over time in 2 and 4 newborns, respectively. Stool homogenates from all newborns elicited elevated inflammatory cytokines, IL-6 and IFN-γ, from mouse macrophages (figure 1). Most newborns were clinically well except for one death from gestational autoimmune liver disease and one with necrotizing enterocolitis. Conclusions These novel findings suggest risk of in utero SARS-CoV-2 transmission to the premature and term fetal intestine during gestation despite negative postnatal nasal PCRs. It is unclear if the presence of viral RNA and protein within the gut microbiome represents active virus in newborns with clinical hospital courses typical of their gestational age in 12 out of 14 cases. However, increasing levels of viral RNA and protein over time suggest replication in some infants, and their gut microbiome induced inflammation in mouse models. The presence of SARS-CoV-2 RNA and spike protein in the intestines of newborns may potentially impact development of the gut microbiome and the immune system and should be further investigated. (Figure Presented).

Endocrine therapy treatment radiosensitizes estrogen receptor-positive breast cancers

Purpose: Estrogen receptor (ER) expression is present in over 80% of breast tumors and has been shown to be a significant driver of breast cancer (BC) pathogenesis and therefore a target of first-line therapies for ER-positive (ER+) BC patients. While both ionizing radiation (RT) and endocrine therapies (ET) are used for the treatment of ER+ BC, the sequencing of therapy and the effect of ET on tumor radiosensitization remain unclear. Recently, this question has become much more clinically relevant when many physicians started offering ET as a bridging strategy to surgery and RT during the COVID-19 pandemic. Here we assessed the efficacy and mechanism of ER inhibition in ER+ BC in combination with RT in preclinical models. Methods: Clonogenic survival assays were used to assess radiosensitization. Inhibition of ER signaling was accomplished by treating ER+ MCF-7 and T47D cells with the selective ER modulator (SERM), tamoxifen, or the selective ER degrader (SERD), fulvestrant. The ER-negative SUM-159 cells were used as a negative control. DNA damage was assessed by the neutral comet assay. Efficiency of homologous recombination (HR) was measured by Rad51 foci or a GFP reporter system. Non-homologous end joining (NHEJ) efficiency was assessed with a pEYFP reporter. Cell cycle effects were measured using flow cytometry with propidium iodide (PI) staining. Apoptosis was assessed by annexin V/PI via flow Scytometry. Senescence was measured using β-galactosidase staining. Western blotting was used to quantify expression of proteins and phospho-proteins involved in cell cycle and apoptosis. An MCF-7 xenograft model was used to assess the efficacy of tamoxifen with RT in vivo. Synergy was determined using the fractional tumor volume (FTV) method. Results: ER inhibition with tamoxifen radiosensitized ER+ MCF-7 (10-250 nM, enhR: 1.14-1.50) and T47D (500 nM-2.0 μ M, enhR: 1.33-1.60) cells but not ER-negative SUM-159 cells (500 nM-2.0 μ M, enhR: 0.99-1.02). ER degradation with fulvestrant had similar radiosensitization effects in MCF-7 (1-25 nM, enhR: 1.33-1.76) and T47D cells (0.5-5 nM, enhR: 0.97-2.81) with no radiosensitization observed in SUM-159 cells (1-25 nM, enhR: 1.01-1.03). MCF-7 cells treated with 500 nM tamoxifen and 4 Gy RT had an increase in dsDNA breaks compared to RT alone as measured by the comet assay (p<0.05), and there was a decrease in NHEJ-mediated repair with tamoxifen treatment (p<0.05). No changes were observed in HR-mediated repair by Rad51 foci or an HR reporter (p=NS). RT alone and in combination with tamoxifen and fulvestrant induced similar levels of cell cycle arrest, suggesting that radiosensitization with the combination therapy is a cell-cycle independent effect. In addition, there were no significant changes in apoptosis in MCF-7 or T47D cells with endocrine therapy, RT, or the combination (p=NS). Although treatment with ET did induce senescence in ER+ MCF-7 and T47D cells, the combination treatment of ET with RT induced senescence to a much greater level suggesting this mechanism may contribute to radiosensitization (p<0.05). In vivo, combination RT and tamoxifen led to a significant delay in time to tumor doubling (17 days in control, 40 days with tamoxifen alone, 32 days with RT alone, and undefined with combination;p<0.0001) and a significant difference in tumor growth between mice treated with tamoxifen or RT alone compared to mice treated with tamoxifen and RT with synergy noted with combination treatment (FTV 1.297). Conclusion: Our data suggest that ET can radiosensitize ER+ breast tumors, and ET with RT may be more effective for radiosensitization. Ongoing studies will address concurrent versus sequential ET with RT. This work also supports further clinical investigation of the timing of RT for patients receiving ET, especially as ET prior to RT is increasingly used as a bridging therapy during the COVID-19 pandemic.

SARS-CoV-2 Ion Channel ORF3a Enables TMEM16F-Dependent Phosphatidylserine Externalization to Augment Procoagulant Activity of the Tenase and Prothrombinase Complexes

Severe SARS-CoV-2 infection is complicated by dysregulation of the blood coagulation system and high rates of thrombosis, but virus-intrinsic mechanisms underlying this phenomenon are poorly understood. Increased intracellular calcium concentrations promote externalization of phosphatidylserine (PS), the membrane anionic phospholipid required for assembly and activation of the tenase and prothrombinase complexes to drive blood coagulation. TMEM16F is a ubiquitous phospholipid scramblase that mediates externalization of PS in a calcium-dependent manner. As SARS-CoV-2 ORF3a encodes a presumed cation channel with the ability to transport calcium, we hypothesized that ORF3a expression by infected host cells perturbs the cellular calcium rheostat, driving TMEM16F-dependent externalization of PS and enhancing procoagulant activity. Using a doxycycline-inducible system, synchronized expression of ORF3a in A549 pulmonary epithelial cells resulted in a time-dependent augmentation of tissue factor (TF) procoagulant activity exceeding 9-fold by 48 hours (p < 0.0001), with no change in TF cell-surface expression. This enhancement was dependent upon PS as determined by inhibition with the PS-binding protein lactadherin. Over 2-fold enhancement of prothrombinase activity (p < 0.0001) was also observed by 48 hours. ORF3a increased intracellular calcium levels by 18-fold at 48 hours (p < 0.0001), as determined by the intracellular calcium indicator fluo-4. After 16 hours of ORF3a expression, more than 60% of cells had externalized PS (p < 0.001) without increased cell death, as quantified by flow cytometry following annexin V binding. Immunofluorescence microscopy staining for ORF3a, annexin V, and nuclei confirmed ORF3a expression within internal and cell surface membranes and increased PS externalization. PS externalization was insensitive to the pan-caspase inhibitor z-VAD-FMK, and there was no evidence of apoptotic activation as determined by caspase-3 cleavage. By contrast, ORF3a expression did not augment coagulation in cells deficient in the calcium-dependent phospholipid scramblase TMEM16F. Similarly, ORF3a-enhanced TF procoagulant activity (p < 0.01) and prothrombinase activity (p<0.05) was completely abrogated using TMEM16 inhibitors, including the uricosuric agent benzbromarone that has been registered for human use in over 20 countries. Live SARS-CoV-2 infection of A549-ACE2 cells increased cell surface factor Xa generation at MOI 0.1 (p < 0.01) but not MOI 0.01 or following heat inactivation of the virus, and RNA sequencing confirmed ORF3a induction without increased F3 expression. RNA sequencing of human SARS-CoV-2 infected lung autopsy and control tissue (n= 53) confirmed these findings in vivo. Immunofluorescence staining for ORF3a and KRT8/18 and CD31 in SARS-CoV-2 infected human lung autopsy specimens demonstrated ORF3a expression in pulmonary epithelium and endothelial cells, highlighting the potential pathologic relevance of this mechanism. Here we demonstrate that expression of the SARS-CoV-2 accessory protein ORF3a increases the intracellular calcium concentration and TMEM16F-dependent PS scrambling to augment procoagulant activity of the tenase and prothrombinase complexes. Our studies of human cells and tissues infected with SARS-CoV-2 support the pathologic relevance of this mechanism. We highlight the therapeutic potential to target the ORF3a-TMEM16F axis as with benzbromarone to mitigate dysregulation of coagulation and thrombosis during severe SARS-CoV-2 infection. Disclosures: Schwartz: Miromatrix Inc: Membership on an entity's Board of Directors or advisory committees;Alnylam Inc.: Consultancy, Speakers Bureau. Schulman: CSL Behring: Consultancy, Research Funding.

Inhibition of Estrogen Receptor Signaling as a Strategy for Radiosensitization of ER+ Breast Cancers

Purpose/Objective(s): Estrogen receptor (ER) expression is present in over 80% of breast tumors and has been shown to be a significant driver of tumor initiation and progression and therefore a target of first-line therapies for ER-positive (ER+) breast cancer (BC) patients. While both ionizing radiation (RT) and endocrine therapies (ET) are used for the treatment of women with ER+ BC, the sequencing of therapy and the effect of ET on tumor radiosensitization remains unclear. Here we assessed the efficacy and mechanism of ER inhibition in ER+ BC in combination with RT in multiple preclinical models. Materials/Methods: Clonogenic survival assays were used to assess radiosensitization. ER inhibition was accomplished with tamoxifen and the selective ER degrader (SERD), fulvestrant, in ER+ MCF-7 and T47D cells or ER-negative (ER-) SUM-159 cells. DNA damage was assessed by yH2AX foci. Efficiency of homologous recombination (HR) or non-homologous end joining (NHEJ) was measured by RAD51 foci or using a pYFP reporter, respectively. Cell cycle effects were measured using flow cytometry with propidium iodide (PI) staining. Apoptosis was assessed by annexin V/PI via flow cytometry. Western blotting was used to quantify protein expression. An MCF-7 xenograft model was used to assess the efficacy of tamoxifen with RT in vivo where mice were pretreated with tamoxifen for one or six days prior to starting RT. Synergy was determined using the fractional tumor volume method. Results: One-hour pretreatment with tamoxifen prior to RT resulted in radiosensitization of ER+ MCF-7 (enhR: 1.14-1.50) and T47D (enhR: 1.33-1.60) cells but not ER- SUM-159 cells (enhR: 0.99-1.02). MCF-7 and T47D cells treated with tamoxifen and RT did not alter the kinetics of dsDNA break repair as measured by yH2AX foci (P > 0.05) but demonstrated a decrease in NHEJ-mediated repair (P < 0.05). No changes were observed in HR-mediated repair by Rad51 foci (P > 0.05). While cell cycle arrest was induced at 24 hours after RT, no changes were observed with tamoxifen treatment in combination with RT. In addition, there were no significant changes in apoptosis in MCF-7 or T47D cells with treatment of tamoxifen, RT, or the combination (P > 0.05). In vivo, an MCF-7 xenograft study demonstrated a significant delay in time to tumor doubling (17 days in control, 40 days with tamoxifen, 32 days with RT, and undefined with combination;P < 0.0001) and a significant difference in tumor growth between mice treated with tamoxifen or RT alone compared to mice treated with tamoxifen and RT with synergy noted with combination treatment. Conclusion: Our data suggest that endocrine therapies may be effectively used to radiosensitize ER+ breast tumors. This work also supports further clinical investigation of the timing of RT for patients receiving ET as concurrent use may be more effective than sequential, especially as ET prior to RT is increasingly used as a bridging therapy during the COVID pandemic.

Initial Single-Center ST-Segment Elevation Myocardial Infarction Experience in New York Before and During the COVID-19 Pandemic

Background: Following a period of stability, the coronavirus disease 2019 (COVID-19) pandemic appears to be re-intensifying globally. As the pandemic continues to evolve, so does our understanding of its implications on ST-segment elevation myocardial infarction (STEMI). We sought to describe a single center STEMI experience at one of the epicenters of the COVID-19 pandemic. Methods: This was a retrospective observational study which included consecutive suspected STEMI patients from March 1 through August 31, 2019, (Cohort 1) compared to the same time period in 2020 (Cohort 2), at a tertiary referral center in Nassau County, New York. Results: Cohort 2 (n=93) saw a similar number of acute myocardial infarction (AMI) team activations compared to cohort 1 (n=90) (Figure 1). Infection control measures and additional investigation were required to clarify the diagnosis in cohort 2, resulting in longer door-to-balloon times (95.9 minutes vs. 74.4 minutes, p=0.0587). We observed similar inpatient length of stay (LOS) (3.6 days vs. 5.0 days, p=0.0901) and mortality (13.2% vs. 9.2%, p=0.5876). Conclusions: Our single-center study, located at one of the epicenters of the pandemic, demonstrated a similar number of AMI team activations, mimicking the seasonal variability seen in 2019, but with longer door-to-balloon times. Despite this, inpatient LOS and mortality remained unchanged. [Formula presented]

COVID-19: The skin and more

The COVID-19 pandemic represents a systemic respiratory infection often with dermatologic signs and systemic sequelae, a public health challenge paralleling the two great influenza pandemics of the last century. COVID-19 have been cutaneous findings are grouped into 6 categories with three distinct indicative patterns: vesicular (varicella-like), vasculopathic, and chilblains-like (including ,COVID toes and ,COVID fingers) plus three less suggestive ones: dermatitic, maculopapular, and urticarial morphologies. Vasculopathic changes are the most concerning, in some patients reflecting a devastating blood clotting dysfunction. Other skin alterations represent adverse cutaneous drug reactions, such as generalized pustular figurate erythema from hydroxychloroquine, and other alterations associated with COVID-19 lifestyle modifications for patients and health care workers.

Nafld and the ymca diabetes prevention program: Repurposing an effective lifestyle intervention program for treatment of nonalcoholic fatty liver disease

Background: Modest weight loss (5-7%) remains the goldstandard of treatment for nonalcoholic fatty liver disease (NAFLD), though patients rarely achieve this independently The YMCA's Diabetes Prevention Program (DPP) is a nationwide structured lifestyle intervention program that has demonstrated weight loss within this range The goal of this study is to determine if the DPP can be repurposed to induce weight loss, decrease liver enzymes, and improve hepatic steatosis in NAFLD Methods: Eligible patients had NAFLD (defined by Fibroscan controlled attenuation parameter (CAP) score >238dB/m) and elevated liver enzymes (ALT>25U/L in females, >33U/L in males) without competing etiologies for liver disease Patients with cirrhosis were excluded A retrospective cohort that received standard of care recommendations was analyzed for change in ALT Two sequential prospective cohorts of 20 patients were enrolled in the experimental YMCA DPP Classroom sessions occurred weekly for 16 weeks, bi-weekly for 4 weeks, and then monthly for 48 weeks total Primary endpoints included change in BMI, ALT, and Fibroscan score. Completers were defined as those attending at least 9 of the first 16 weekly sessions. Changes in variables were analyzed using paired t-test End of study data was not completed for cohort 2 due to COVID19 and therefore aggregate data is presented for week 16 and end of study data (week 48) is presented for only cohort 1 Results: A total of 20 patients from both cohorts were completers 16% were female (n=12) 16 patients had interim study data and were included in the week 16 analysis All but one patient lost weight Baseline mean BMI was 34 3 and decreased to 33 6 by week 16 (p=0 02) Baseline mean ALT was 91 8U/L and decreased to 55 1U/L by week 16 (p=0 005) Mean Fibroscan fibrosis score improved from 6.7kPa to 5.5kPa and CAP score improved from 321dB/m to 291dB/m by week 16 (p=0 008 and p=0 12, respectively) 9 patients from cohort 1 were included in the week 48 analysis Mean BMI was 35 4 at baseline and decreased to 33 6 by week 48 (p=0 002) Mean ALT was 85 0U/L at baseline decreased to 41 6U/L (p=0 01) Mean Fibroscan score improved from 6 9kPa to 5 9kPa (p=0 07) Other metabolic parameters also trended towards significant improvement at week 16 and 48 (Table 1). The retrospective cohort (N=31) had a mean baseline ALT of 69U/L with no significant change at 6-month follow-up (mean ALT 60U/L, p=0 24) Conclusion: Standard of care lifestyle recommendations are ineffective in inducing weight loss and improvement in ALT in NAFLD The YMCA DPP repurposed to treat NAFLD resulted in statistically significant improvements in BMI and ALT in patients with NAFLD Fibroscan scores and other metabolic parameters also improved, though the sample size is likely too small to see a statistically significant improvement The existing infrastructure established by the YMCA throughout the U S could emerge as a leading treatment option for NAFLD patients.

Human pluripotent stem cell derived hepatic platforms to study SARS-COV-2 infection

Background: COVID-19 was declared a pandemic by the World Health Organization, caused by severe acute respiratory syndrome corona-virus 2 (SARS-CoV-2) Respiratory failure is the most common mortality outcome in COVID-19 patients, yet serious and even fatal manifestations are seen across multiple organ systems Emerging clinical data from our own hospital revealed a high prevalence of initial presentations with GI manifestations Almost one third of patients presenting to our hospital reported at least one gastrointestinal manifestation including nausea, vomiting, diarrhea, or abdominal pain 62% of patients presented with biochemical evidence of liver injury Moreover, the presence of liver injury on presentation was associated with a significantly higher risk of ICU admission and death As this is a new and novel clinical entity, robust in vitro models that phenocopy SARS-CoV-2 infection and human COVID-19 disease are limited Current in vitro (e g Vero cells) and in vivo models (mouse models engineered with ACE2) are so distinct from human infection that they may not capture key components of viral infection or virus-host interactions Therefore, the development of robust human models of COVID-19 infection will be essential for the study of SARS-Cov-2 viral infection and to identify robust SARS-CoV-2 therapeutics Methods: Human pluripotent stem cells (hPSCs), including human embryonic stem cells hESCs) and induced pluripotent stem cells (hiPSCs), can be used to derive functional human cells/tissues/organoids for modeling human disease and drug discovery, including for infectious diseases Here we leveraged several stem cell platforms (e g endodermal lineages including hepatocyte and cholangiocyte) along with primary human hepatocytes and cholangiocyte organoid systems to study SARS-CoV-2 infection SARS-CoV-2 pseudoparticles were used to study SARS-CoV-2 viral entry SARS-CoV-2 (USA-WA1/2020) was used to validate viral infection and to study cellular response Autopsy liver samples from COVID-19 patients were obtained and compared to SARS-CoV-2 infected liver models Results: Adult hepatocyte and cholangiocyte organoids along with PSC derived hepatocytes and cholangiocytes are Permissive to SARS-CoV-2 virus infection and show similar transcriptome changes and chemokine responses for SARSCoV- 2 infection as seen in autopsy samples from COVID-19 Patients Conclusion: We report here the development of robust models of SARS-CoV-2 infection in primary and PSC derived hepatocyte and cholangiocytes which phenocopy COVID-19 hepatic disease These disease-relevant human cell/organoid-based platforms can be directly applied for drug screening and the evaluation of prospective antiviral therapeutics as well be used to delineate molecular mechanisms underlying COVID-19 disease.