Targeted Expression to Liver of an antimiR-33 Sponge as a Gene Therapy Strategy against Hypercholesterolemia: In Vitro Study.

Atherosclerosis is the leading cause of cardiovascular diseases in Mexico and worldwide. The membrane transporters ABCA1 and ABCG1 are involved in the reverse transport of cholesterol and stimulate the HDL synthesis in hepatocytes, therefore the deficiency of these transporters promotes the acceleration of atherosclerosis. MicroRNA-33 (miR-33) plays an important role in lipid metabolism and exerts a negative regulation on the transporters ABCA1 and ABCG1. It is known that by inhibiting the function of miR-33 with antisense RNA, HDL levels increase and atherogenic risk decreases. Therefore, in this work, a genetic construct, pPEPCK-antimiR-33-IRES2-EGFP, containing a specific antimiR-33 sponge with two binding sites for miR-33 governed under the PEPCK promoter was designed, constructed, and characterized, the identity of which was confirmed by enzymatic restriction, PCR, and sequencing. Hep G2 and Hek 293 FT cell lines, as well as a mouse hepatocyte primary cell culture were transfected with this plasmid construction showing expression specificity of the PEPCK promoter in hepatic cells. An analysis of the relative expression of miR-33 target messengers showed that the antimiR-33 sponge indirectly induces the expression of its target messengers (ABCA1 and ABCG1). This strategy could open new specific therapeutic options for hypercholesterolemia and atherosclerosis, by blocking the miR-33 specifically in hepatocytes.

A Portable Triboelectric Nanogenerator Based on Dehydrated Nopal Powder for Powering Electronic Devices.

Triboelectric nanogenerators (TENGs) based on organic materials can harvest green energy to convert it into electrical energy. These nanogenerators could be used for Internet-of-Things (IoT) devices, substituting solid-state chemical batteries that have toxic materials and limited-service time. Herein, we develop a portable triboelectric nanogenerator based on dehydrated nopal powder (NOP-TENG) as novel triboelectric material. In addition, this nanogenerator uses a polyimide film tape adhered to two copper-coated Bakelite plates. The NOP-TENG generates a power density of 2309.98 µW·m-2 with a load resistance of 76.89 MΩ by applying a hand force on its outer surface. Furthermore, the nanogenerator shows a power density of 556.72 µW·m-2 with a load resistance of 76.89 MΩ and under 4g acceleration at 15 Hz. The output voltage of the NOP-TENG depicts a stable output performance even after 27,000 operation cycles. This nanogenerator can light eighteen green commercial LEDs and power a digital calculator. The proposed NOP-TENG has a simple structure, easy manufacturing process, stable electric behavior, and cost-effective output performance. This portable nanogenerator may power electronic devices using different vibration energy sources.

COVID-19 impact on the psychological health of Latinx transgender and non-binary individuals in mainland United States and Puerto Rico: a mixed-methods study.

BACKGROUND: The COVID-19 pandemic continues to generate an unprecedented impact on all aspects of everyday life across the world. However, those with historically and currently marginalized identities (i.e., gender or ethnicity) who already experience a wide range of structural inequities have been disproportionally impacted. LTNB are a particularly at-risk population as they lie at the intersection of race/ethnicity, gender identity, language, migration status, geographical location, among others, which could further increase their COVID-19 and other health-related risks and disparities. The objective of this study was to examine the impact of key social determinants of health (i.e., gender identity, country, health insurance, employment) among a sample of LTNB individuals. METHODS: The team implemented a cross-sectional exploratory design with an online survey technique using the secure web platforms REDcap and SurveyMonkey. A total of 133 participants completed the online survey. Most of the sample self-identified as transwomen (38.8%), transmen (26.3%), and non-binary (21.8%) between the ages of 21 to 72. All participants were Latinx living in either Puerto Rico (47.7%) or mainland United States (52.3%). Descriptive statistics, reliability tests, Mann-Whitney and rapid thematic analysis test were conducted. RESULTS: Findings show that most participants were always (38.1%) or almost always (33.3%) worried about contracting COVID-19. Individuals living in Puerto Rico reported more difficulties than those residing in the mainland US regarding COVID-19 impact on psychosocial, emotional, and COVID-related thinking. Most participants' answers for the COVID-19 open-ended questions focused on three main domains: income, access to trans-affirmative health care, and coping strategies. DISCUSSION: Findings evidence that although most of LTNB participants were negatively impacted by the COVID-19 pandemic in multiple aspects of their lives, those living in Puerto Rico experienced these differently when compared to those in mainland US. More research is needed to understand better the mechanisms and pathways through which this context specifically impacts LTNB health and wellbeing, particularly in Puerto Rico. This study could help shape the public health response taking into account the geographical location and other intersectional identities that play critical roles in the production and reproduction of inequities.

Effects of Bisphenol A on Foxl2 gene expression and DNA damage in adult viviparous fish Goodeaatripinnis.

Bisphenol A (BPA) is an emerging pollutant of global concern. Viviparous fish Goodea atripinnis is endemic to the Central Mexican Plateau where BPA was detected; however, few studies examined the influence of this chemical on native viviparous fish. The effects of BPA (sublethal dose) were determined on DNA integrity and Foxl2 expression in G. atripinnis gonads, and interactions of BPA with FOXL2 protein. Genotoxicity analysis revealed that % comets, at 14 and 28 days and comet tail length (at 14 days) were significantly higher in exposed compared to controls. In general, the % DNA tail was not markedly higher in BPA-treated fish; however, tail moment related to tail length exhibited significant increases in DNA damage. RT-qPCR assays showed Foxl2 overexpression after 14 and 28 days of exposure in females; while in males, Foxl2 was overexpressed after 28 days. In silico analysis demonstrated that BPA interacted with seven residues located in FOXL2 homeodomain. In summary, sublethal BPA doses induced DNA damage and changes in Foxl2 expression in gonadal cells of G. atripinnis, which may adversely affect reproduction in BPA-exposed wild populations. Foxl2 overexpression and BPA-FOXL2 interaction suggested alterations in processes involving Foxl2. Viviparous fish may thus serve as potential non-conventional models for assessing pollutants effects.

Synthesis of ZnO Nanorod Film Deposited by Spraying with Application for Flexible Piezoelectric Energy Harvesting Microdevices.

Industry 4.0 and the Internet of Things have significantly increased the use of sensors and electronic products based on flexible substrates, which require electrical energy for their performance. This electrical energy can be supplied by piezoelectric vibrational energy harvesting (pVEH) devices. These devices can convert energy from ambient mechanical excitations into electrical energy. In order to develop, these devices require piezoelectric films fabricated with a simple and low-cost process. In this work, we synthesize ZnO nanorod film by a solvothermal method and deposit by spraying on ITO (indium-tin-oxide)/PET (polyethylene terephthalate) flexible substrate for a pVEH microdevice. The results of the characterization of the ZnO nanorod film using X-ray diffraction (XRD) confirm the typical reflections for this type of nanomaterial (JCPDS 36-145). Based on transmission electron microscopy (TEM) images, the size of the nanorod film is close to 1380 nm, and the average diameter is 221 ± 67 nm. In addition, the morphological characteristics of the ZnO nanorod film are obtained using atomic force microscopy (AFM) tapping images. The pVEH microdevice has a resonant frequency of 37 Hz, a generated voltage and electrical power of 9.12 V and 6.67 µW, respectively, considering a load resistance of 107.7 kΩ and acceleration of 1.5 g. The ZnO nanorod film may be applied to pVEH microdevices with flexible substrates using a low-cost and easy fabrication process.

Complications associated with Pediatric Supracondylar Humeral Fractures.

OBJECTIVE: To measure pediatric supracondylar fracture epidemiology, fracture rate, and complications in the island's pediatric population. The study aims to compare our results to the national rates reported in the literature. METHODS: In this retrospective record review we examined 330 patients who underwent open reduction internal fixation or closed reduction percutaneous pinning in pediatric supracondylar fractures at the University Pediatric Hospital of Puerto Rico Medical Center (HOPU). The study evaluated patients from January 2008 to January 2011 that had completed at least a 1 year follow-up. Measurement of type of fracture, and complications were recorded. Statistical significance was set at a p-valuevalue<0.05. RESULTS: Of the 330 patients, 206 (62%) were male and 127 (38%) were female. The average age was 5.49 years (± 2.43). The vast majority had extension-type fractures (98.2%) and 1.8% had flexion-type fractures. The neurological complication rates were 10% (33 patients). Neurologic complications after distal fragment displacement were 13.5% for posteromedial displacement versus 11.8% for posterolateral displacement, with a p-value of 0.71. CONCLUSION: Our results in terms of female-to-male ratio, fracture type and complications (e.g., infection, vascular and neurologic complications) were similar to those reported in the literature. Because significant differences in the rates of posteromedial and posterolateral supracondylar fractures were found, we recommend further research on this subject. Overall, both our findings regarding complications and our results are similar to what has been reported in the literature. We can therefore affirm that our institution provides adequate care and management for this kind of fracture.

Moderate patient-prosthesis mismatch predicts cardiac events and advanced functional class in young and middle-aged patients undergoing surgery due to severe aortic stenosis.

BACKGROUND AND AIM: The clinical impact of patient-prosthesis mismatch (PPM) on outcomes in young and middle-aged patients undergoing surgery for aortic valve replacement (AVR) remains unknown. Our objective was to examine the mid-term impact of PPM on overall mortality, quality of life, and cardiac events in this population. METHODS: All patients younger than 70 years of age undergoing isolated AVR from October 2005 to October 2011 were analyzed. PPM was defined as the indexed effective orifice area ≤ 0.85 cm(2) /m(2) . We studied the impact of PPM on mid-term overall mortality, cardiac events, and New York Heart Association functional class using an analysis stratified for propensity score. Cardiac events were defined as cardiac death, sudden death, hospital readmission due to angina, syncope or heart failure or reoperation on aortic prosthesis. RESULTS: Two hundred and ninety-three patients were included in the study, of whom 81 (27.61%) had some degree of PPM. PPM had no impact on mid-term overall mortality (HR=1.45; 95% CI=0.65-3.22; p=0.36), although it had a negative impact on cardiac events (HR=11.52; 95% CI=5.25-25.24; p<0.001) and functional class (RR=7.55; 95% CI=2.59-22.03; p<0.001). CONCLUSIONS: Moderate PPM appears to be a strong and independent predictor of cardiac events and advanced functional class in young and middle-aged patients undergoing AVR for severe stenosis. However, it is possible that it has no impact on overall mortality.

Studying SARS-CoV-2 interactions using phage-displayed receptor binding domain as a model protein.

SARS-CoV-2 receptor binding domain (RBD) mediates viral entry into human cells through its interaction with angiotensin converting enzyme 2 (ACE2). Most neutralizing antibodies elicited by infection or vaccination target this domain. Such a functional relevance, together with large RBD sequence variability arising during viral spreading, point to the need of exploring the complex landscape of interactions between RBD-derived variants, ACE2 and antibodies. The current work was aimed at developing a simple platform to do so. Biologically active and antigenic Wuhan-Hu-1 RBD, as well as mutated RBD variants found in nature, were successfully displayed on filamentous phages. Mutational scanning confirmed the global plasticity of the receptor binding motif within RBD, highlighted residues playing a critical role in receptor binding, and identified mutations strengthening the interaction. The ability of vaccine-induced antibodies to inhibit ACE2 binding of many mutated RBD variants, albeit at different extents, was shown. Amino acid replacements which could compromise such inhibitory potential were underscored. The expansion of our approach could be the starting point for a large-scale phage-based exploration of diversity within RBD of SARS-CoV-2 and related coronaviruses, useful to understand structure-function relationships, to engineer RBD proteins, and to anticipate changes to watch during viral evolution.

Carcinogenic effect of human tumor-derived cell-free filtrates in nude mice.

Cancer remains a worldwide cause of morbidity and mortality. Investigational research efforts have included the administration of tumor-derived extracts to healthy animals. Having previously demonstrated that the administration of non-transmissible, human cancer-derived homogenates induced malignant tumors in mice, here, we examined the consequences of administering 50 or 100 µg of protein of crude homogenates from mammary carcinoma, pancreatic adenocarcinoma, and melanoma samples in 6 inoculations per week during 2 months. The concurrent control mice received homogenates of healthy donor-skin cosmetic surgery fragments. Mammary carcinoma homogenate administration did not provoke the deterioration or mortality of the animals. Multiple foci of lung adenocarcinomas with a broad expression of malignity histomarkers coexisting with small cell-like carcinomas were found. Disseminated cells, positive to classic epithelial markers, were detected in lymphoid nodes. The administration of pancreatic tumor and melanoma homogenates progressively deteriorated animal health. Pancreatic tumor induced poorly differentiated lung adenocarcinomas and pancreatic islet hyperplasia. Melanoma affected lungs with solid pseudopapillary adenocarcinomas. Giant atypical hepatocytes were also observed. The kidney exhibited dispersed foci of neoplastic cells within a desmoplastic matrix. Nuclear overlapping with hyperchromatic nuclei, mitotic figures, and prominent nuclear atypia was identified in epidermal cells. None of these changes were ever detected in the control mice. Furthermore, the incubation of zebrafish embryos with breast tumor homogenates induced the expression of c-Myc and HER-2 as tumor markers, contrasting to embryos exposed to healthy tissue-derived material. This study confirms and extends our hypothesis that tumor homogenates contain and may act as vectors for "malignancy drivers," which ultimately implement a carcinogenesis process in otherwise healthy mice.

Eco-friendly, compact, and cost-efficient triboelectric nanogenerator for renewable energy harvesting and smart motion sensing.

In recent years, the growth of Internet of Things devices has increased the use of sustainable energy sources. An alternative technology is offered by triboelectric nanogenerators (TENGs) that can harvest green energy and convert it into electrical energy. Herein, we assessed three different nopal powder types that were used as triboelectric layers of eco-friendly and sustainable TENGs for renewable energy harvesting from environmental vibrations and powering electronic devices. These nanogenerators were fabricated using waste and recycled materials with a compact design for easy transportation and collocation on non-homogeneous surfaces of different vibration or motion sources. In addition, these TENGs have advantages such as high output performance, stable output voltage, lightweight, low-cost materials, and a simple fabrication process. These nanogenerators use the contact-separation mode between two triboelectric layers to convert the vibration energy into electrical energy. TENG with the best output performance is based on dehydrated nopal powder, generating an output power density of 2.145 mWm-2 with a load resistance of 39.97 MΩ under 3g acceleration and 25 Hz operating frequency. The proposed TENGs have stable output voltages during 22500 operating cycles. These nanogenerators can light 116 ultra-bright blue commercial LEDs and power a digital calculator. Also, the TENGs can be used as a chess clock connected to a mobile phone app for smart motion sensing. These nanogenerators can harvest renewable vibration energy and power electronic devices, sensors, and smart motion sensing.

Review of the psychometric properties of the Patient Health Questionnaire-9 (PHQ-9) Spanish version in a sample of Puerto Rican workers.

Background: This study aimed to examine the internal structure and assess the psychometric properties of the Patient Health Questionnaire (PHQ-9) in a Puerto Rican sample of workers. This instrument is a nine-item questionnaire, which was conceptualized as a unidimensional structure; however, there are mixed results regarding this internal structure. This measure is used in the occupational health psychology context in organizations in Puerto Rico; nevertheless, there is little evidence of its psychometric properties with samples of workers. Materials and methods: A total of 955 samples from two different study samples were used in this cross-sectional study design in which the PHQ-9 was used. We conducted confirmatory factor analysis, bifactor analysis, and random intercept item factor analysis to examine the internal structure of the PHQ-9. Moreover, a two-factor model was examined by randomly assigning items to the two factors. Measurement invariance across sex and the relationship with other constructs were examined. Results: The best-fitted model was the bifactor model followed by the random intercept item factor. The five sets of two-factor models with items randomly assigned obtained acceptable and similar fit indices regardless of the items. Conclusion: The results suggest that the PHQ-9 appears to be a reliable and valid instrument to measure depression. The more parsimonious interpretation of its scores, for now, is a unidimensional structure. Comparison across sex appears to be useful in occupational health psychology research settings since the results suggest that the PHQ-9 is invariant regarding this variable.

Psychometric properties of the generalized anxiety disorder-7 (GAD-7) in a sample of workers.

Objective: To evaluate the psychometric properties of the GAD-7 by obtaining evidence of internal structure (dimensionality, precision and differential functioning of items) and association with external variables. Methods: A total of 2,219 protocols from three different studies conducted with Puerto Rican employees that administered the GAD-7 were selected for the current study. Item response theory modeling was used to assess internal structure, and linear association with external variables. Results: The items were adapted to a graduated response model, with high similarity in the discrimination and location parameters, as well as in the precision at the level of the items and in the total score. No violation of local independence and differential item functioning was detected. The association with convergent (work-related rumination) and divergent (work engagement, sex, and age) variables were theoretically consistent. Conclusion: The GAD-7 is a psychometrically robust tool for detecting individual variability in symptoms of anxiety in workers.

Diagnostic Hesitancy of Primary Care Physicians in Puerto Rico Toward Alzheimer's Disease and Related Dementias: Opportunities for Transformation.

OBJECTIVE: Migration of physicians in Puerto Rico makes it difficult to obtain specialized care for Alzheimer's disease and related dementias (ADRD). Primary care physicians (PCPs) can fill this gap, but there is limited information on how their beliefs and behaviors towards ADRD affect early diagnostic practices. Using the Theory of Planned Behavior (TPB), we addressed salient beliefs of PCPs that affect their intention to diagnose ADRD early, defined as "performing the recommended clinical testing to diagnose ADRD within 3 months from the patient's first subjective memory complaint". METHODS: This cross-sectional study included 103 Puerto Rican PCPs, surveyed at CME activities and online. We measured PCPs salient beliefs in terms of attitudes, pressure they feel from others, self-perception of ability to diagnose ADRD early, and intention to perform early diagnosis. Questionnaire measures were psychometrically acceptable. RESULTS: Structural equation modeling (PLS-SEM) analysis showed that, combined, framework constructs explained 35.7% of the variance, reflecting a moderate-to-strong intention of PCPs to diagnose early (R2 = 0.357, p < 0.001). Self-perception of skills was the strongest predictor of intention (ß= 0.378, p < 0.001). Hours in ADRD training and years of experience in medical practice were strongly correlated with the percentage of total diagnoses performed by PCPs. CONCLUSION: Training and years of experience may be key for PCPs to have a positive outlook of their skills for early ADRD diagnosis. Findings could be useful to design interventions to dispel myths about ADRD, reduce stigma, and reduce diagnostic hesitancy toward ADRD among PCPs.

[Takotsubo cardiomyopathy associated with abnormal birth of the coronary arteries]. / Miocardiopatía de Takotsubo asociada al nacimiento anómalo de las arterias coronarias.

Background: Takotsubo cardiomyopathy (TM) is a form of non-ischemic cardiomyopathy. It is characterized by transient regional systolic dysfunction of the left ventricle that mimics acute myocardial infarction. The main objective of this article is to report the case of a patient with TM associated with abnormal birth of the left coronary trunk. Clinical case: A 76-year-old woman with typical angina at rest, with an electrocardiogram that showed dynamic changes in the T wave and elevation of biomarkers. Coronary angiography showed epicardial coronary arteries without significant lesions and abnormal birth of the left coronary trunk from the proximal segment of the right coronary artery. The patient progressed favorably, and the transthoracic echocardiogram showed no mobility disorders 3 months after the event. Conclusions: TM and abnormal birth of the coronary arteries are rare diseases whose simultaneous presentation is extraordinary. The diagnosis of both clinical entities is made by coronary angiography and echocardiogram, and their treatment is similar to that of patients with acute coronary syndrome. Abnormal birth of the left coronary trunk with retroaortic switch reaching the contralateral site has a good clinical prognosis and echocardiographic follow-up should be performed 4 weeks after the onset of the condition.

Introducción: la miocardiopatía de Takotsubo (MT) es una forma de miocardiopatía no isquémica. Se caracteriza por la disfunción sistólica regional transitoria del ventrículo izquierdo que imita al infarto agudo de miocardio. El objetivo principal de este artículo es reportar el caso de una paciente con MT asociada al nacimiento anómalo del tronco coronario izquierdo. Caso clínico: paciente mujer de 76 años que presentó angina típica en reposo, con un electrocardiograma que evidenció cambios dinámicos en la onda T y elevación de biomarcadores. La coronariografía evidenció a las arterias coronarias epicárdicas sin lesiones significativas y el nacimiento anómalo del tronco coronario izquierdo proveniente del segmento proximal de la arteria coronaria derecha. La paciente evolucionó de manera favorable y el ecocardiograma transtorácico no mostró trastornos en la movilidad a los tres meses del evento. Conclusiones: la MT y el nacimiento anómalo de las arterias coronarias son enfermedades raras cuya presentación simultánea es extraordinaria. El diagnóstico de ambas entidades clínicas se realiza mediante la coronariografía y el ecocardiograma, y su tratamiento es similar al de los pacientes con síndrome coronario agudo. El nacimiento anómalo del tronco coronario izquierdo con cruce retroaórtico que alcanza el sitio contralateral tiene un buen pronóstico y se debe realizar seguimiento clínico y ecocardiográfico a las cuatro semanas del inicio del padecimiento.

The impact of the Covid-19 pandemic and previous natural disasters on the mental health of healthcare workers in Puerto Rico.

The objective of this study was to assess the impact of COVID-19 pandemic worries (e.g., fear of contagion) and previous exposure to natural disasters (e.g., hurricanes) on Healthcare Workers (HCWs) mental health in Puerto Rico. Participants completed a self-administered online survey including items on sociodemographic information, working conditions, fears, and worries about the COVID-19 pandemic, past natural disaster experiences, depressive symptoms, and resilience. Logistic regressions models were performed to explain the relationship between depressive symptomatology and COVID-19 experiences and worries. 40.9% (n = 107) of the sample were classified as having some level (mild to severe) of depressive symptomatology (PHQ-8 ≥5). Results reflect normal to high levels of psychological resilience (BRS; M = 3.7, SD = 0.7). A significant association was found between depressive symptomatology and psychological resilience (OR = 0.44, 95% CI: 0.25-0.77). The odds of having depressive symptomatology were almost five times higher (OR = 4.79, 95% CI: 1.71-13.44) among those who reported emotional coping difficulties during the pandemic after experiencing a natural disaster compared to those that did not, when adjusting for psychological resilience and residence region. Despite normal to high psychological resilience levels, HCWs who reported emotional coping difficulties due to previous disasters were at risk of developing depressive symptomatology. Results suggest interventions to address the mental health of HCWs could benefit from considering other individual and environmental factors beyond resilience. Findings could inform future interventions to promote HCWs' well-being before, during, and after a natural disaster or pandemic outbreak.

Impact of plasma-activated water on the supramolecular structure and functionality of small and large starch granules.

Hydrothermal (HMT) and water agitation (WA) treatments using plasma-activated water (PAW) were employed as sustainable methods to modify the molecular and functional performance of small (rice) and large (potato) starch granules. HMT-PAW and WA-PAW treatments resulted in etched and damaged granular surfaces that rearranged the long and short-range crystallinity of the modified starches. Both treatments seemed to predominantly occur in the amorphous region of the rice starch and the crystalline regions of the potato starch, changing the crystallinity values from 22.9 and 14.8 % to 31.8 and 10.4 %, respectively. Thus, the level of the arrangement of chains reached after PAW treatment decreased the ability of rice starch granules to swell (16 to 9 %) and leach out starch molecules from the granules (4.5 to 1.3 %), decreasing the viscosity and pasting profiles as indicated by n and k values. Opposite behavior was observed in the modified potato starches since starch components leached out to a higher extent (1.7 to 5.4 %). The results showed that HMT and WA treatments using PAW are feasible eco-friendly methods for modifying starch granules without chemical reagents. These modified starches could be suitable as functional ingredients or biopolymeric matrices for the food and packaging industry.

Molecular reshaping of phage-displayed Interleukin-2 at beta chain receptor interface to obtain potent super-agonists with improved developability profiles.

Interleukin-2 (IL-2) engineered versions, with biased immunological functions, have emerged from yeast display and rational design. Here we reshaped the human IL-2 interface with the IL-2 receptor beta chain through the screening of phage-displayed libraries. Multiple beta super-binders were obtained, having increased receptor binding ability and improved developability profiles. Selected variants exhibit an accumulation of negatively charged residues at the interface, which provides a better electrostatic complementarity to the beta chain, and faster association kinetics. These findings point to mechanistic differences with the already reported superkines, characterized by a conformational switch due to the rearrangement of the hydrophobic core. The molecular bases of the favourable developability profile were tracked to a single residue: L92. Recombinant Fc-fusion proteins including our variants are superior to those based on H9 superkine in terms of expression levels in mammalian cells, aggregation resistance, stability, in vivo enhancement of immune effector responses, and anti-tumour effect.

Development of a scalable single process for producing SARS-CoV-2 RBD monomer and dimer vaccine antigens.

We have developed a single process for producing two key COVID-19 vaccine antigens: SARS-CoV-2 receptor binding domain (RBD) monomer and dimer. These antigens are featured in various COVID-19 vaccine formats, including SOBERANA 01 and the licensed SOBERANA 02, and SOBERANA Plus. Our approach involves expressing RBD (319-541)-His6 in Chinese hamster ovary (CHO)-K1 cells, generating and characterizing oligoclones, and selecting the best RBD-producing clones. Critical parameters such as copper supplementation in the culture medium and cell viability influenced the yield of RBD dimer. The purification of RBD involved standard immobilized metal ion affinity chromatography (IMAC), ion exchange chromatography, and size exclusion chromatography. Our findings suggest that copper can improve IMAC performance. Efficient RBD production was achieved using small-scale bioreactor cell culture (2 L). The two RBD forms - monomeric and dimeric RBD - were also produced on a large scale (500 L). This study represents the first large-scale application of perfusion culture for the production of RBD antigens. We conducted a thorough analysis of the purified RBD antigens, which encompassed primary structure, protein integrity, N-glycosylation, size, purity, secondary and tertiary structures, isoform composition, hydrophobicity, and long-term stability. Additionally, we investigated RBD-ACE2 interactions, in vitro ACE2 recognition of RBD, and the immunogenicity of RBD antigens in mice. We have determined that both the monomeric and dimeric RBD antigens possess the necessary quality attributes for vaccine production. By enabling the customizable production of both RBD forms, this unified manufacturing process provides the required flexibility to adapt rapidly to the ever-changing demands of emerging SARS-CoV-2 variants and different COVID-19 vaccine platforms.

Stigma and social support and their impact on quality of life and self-esteem among women with endometriosis in Latin-America and the Caribbean.

Endometriosis is a complex disease affecting approximately 5-10% individuals worldwide. Prevalence rates in Latin-America and the Caribbean are largely unknown, with published data only from Puerto Rico and Chile. Pain associated with endometriosis negatively affects patients' health and quality of life. However, there is a need to better understand the role played by psychosocial variables such as stigma and social support in diverse cultural contexts. The purpose of this study was to examine the mediating role of endometriosis related stigma (endo-stigma) and the moderating role of social support on the endometriosis QoL and self-esteem among women with endometriosis from Latin America and the Caribbean. A cross-sectional design with online survey techniques was implemented. A total of 169 self-identified cisgender women with endometriosis from 14 Latin-American and Caribbean countries participated in the study. We used partial least squares structural equation modeling (PLS-SEM) to examine the study's hypotheses. Incapacitating pain was positively and significantly related to endometriosis QoL as measured by the EHP-5 (b = .266, p < .01). Endo-stigma was positively and significantly related to endometriosis QoL (b = .340, p< .01) and self-esteem (b = .297, p< .01). In addition, endo-stigma mediated the relationship between incapacitating pain and self-esteem (IE = .073, p = .018). Finally, social support moderated the relationship between stigma stress and endometriosis QoL (b = .060, p = .039). Findings suggest stigma could be one of the mechanisms through which the relationship between incapacitating pain and self-esteem among Latin American and Caribbean women with endometriosis could be partially explained. Furthermore, women who scored high in the need for social support and stigma stress also showed worst endometriosis QoL. These results point towards the need to develop tailored interventions targeting these factors in order to foster a better QoL and wellbeing for this population in the context of Latin America and the Caribbean.

COVID-19 impact on the psychological health of Latinx transgender and non-binary individuals in mainland United States and Puerto Rico: A mixed-methods study.

Background The COVID-19 pandemic continues to generate an unprecedented impact on all aspects of everyday life across the world. However, those with historically and currently marginalized identities (i.e., gender or ethnicity) who already experience a wide range of structural inequities have been disproportionally impacted. LTNB are a particularly at-risk population as they lie at the intersection of race/ethnicity, gender identity, language, migration status, geographical location, among others, which could further increase their COVID-19 and other health-related risks and disparities. The objective of this study was to examine the impact of key social determinants of health (i.e., gender identity, country, health insurance, employment) among a sample of LTNB individuals. Methods The team implemented a cross-sectional exploratory design with an online survey technique using the secure web platforms REDcap and SurveyMonkey. A total of 133 participants completed the online survey. Most of the sample self-identified as transwomen (38.8%), transmen (26.3%), and non-binary (21.8%) between the ages of 21 to 72. All participants were Latinx living in either Puerto Rico (47.7%) or mainland United States (52.3%). Descriptive statistics, reliability tests, Mann-Whitney and rapid thematic analysis test were conducted. Results Findings show that most participants were always (38.1%) or almost always (33.3%) worried about contracting COVID-19. Individuals living in Puerto Rico reported more difficulties than those residing in the mainland US regarding COVID-19 impact on psychosocial, emotional, and COVID-related thinking. Most participants' answers for the COVID-19 open-ended questions focused on three main domains: income, access to trans-affirmative health care, and coping strategies. Discussion Findings evidence that although most of LTNB participants were negatively impacted by the COVID-19 pandemic in multiple aspects of their lives, those living in Puerto Rico experienced these differently when compared to those in mainland US. More research is needed to understand better the mechanisms and pathways through which this context specifically impacts LTNB health and wellbeing, particularly in Puerto Rico. This study could help shape the public health response taking into account the geographical location and other intersectional identities that play critical roles in the production and reproduction of inequities.