Membrane potential hyperpolarization: a critical factor in acrosomal exocytosis and fertilization in sperm within the female reproductive tract.

Hyperpolarization of the membrane potential (Em), a phenomenon regulated by SLO3 channels, stands as a central feature in sperm capacitation-a crucial process conferring upon sperm the ability to fertilize the oocyte. In vitro studies demonstrated that Em hyperpolarization plays a pivotal role in facilitating the mechanisms necessary for the development of hyperactivated motility (HA) and acrosomal exocytosis (AE) occurrence. Nevertheless, the physiological significance of sperm Em within the female reproductive tract remains unexplored. As an approach to this question, we studied sperm migration and AE incidence within the oviduct in the absence of Em hyperpolarization using a novel mouse model established by crossbreeding of SLO3 knock-out (KO) mice with EGFP/DsRed2 mice. Sperm from this model displays impaired HA and AE in vitro. Interestingly, examination of the female reproductive tract shows that SLO3 KO sperm can reach the ampulla, mirroring the quantity of sperm observed in wild-type (WT) counterparts, supporting that the HA needed to reach the fertilization site is not affected. However, a noteworthy distinction emerges-unlike WT sperm, the majority of SLO3 KO sperm arrive at the ampulla with their acrosomes still intact. Of the few SLO3 KO sperm that do manage to reach the oocytes within this location, fertilization does not occur, as indicated by the absence of sperm pronuclei in the MII-oocytes recovered post-mating. In vitro, SLO3 KO sperm fail to penetrate the ZP and fuse with the oocytes. Collectively, these results underscore the vital role of Em hyperpolarization in AE and fertilization within their physiological context, while also revealing that Em is not a prerequisite for the development of the HA motility, essential for sperm migration through the female tract to the ampulla.

Protection of homologous and heterologous boosters after primary schemes of rAd26-rAd5, ChAdOx1 nCoV-19 and BBIBP-CorV during the omicron outbreak in adults of 50 years and older in Argentina: a test-negative case-control study.

Background: After primary vaccination schemes with rAd26-rAd5 (Sputnik V), ChAdOx1 nCoV-19, BBIBP-CorV or heterologous combinations, the effectiveness of homologous or heterologous boosters (Sputnik V, ChAdOx, Pfizer-BioNTech, Moderna) against SARS-CoV-2 infections, hospitalisations and deaths has been scarcely studied. Methods: Test-negative, case-control study, conducted in Argentina during omicron BA.1 predominance, in adults ≥50 years old tested for SARS-CoV-2 who had received two or three doses of COVID-19 vaccines. Outcomes were COVID-associated infections, hospitalisations and deaths after administering mRNA and vectored boosters, < or ≥60 days from the last dose. Findings: Of 422,124 individuals tested for SARS-CoV-2, 221,993 (52.5%) tested positive; 190,884 (45.2%) and 231,260 (54.8%) had received 2-dose and 3-dose vaccination schemes, respectively. The 3-dose scheme reduced infections, hospitalisations and death (OR 0.81 [0.80-0.83]; 0.28 [0.25-0.32] and 0.25 [0.22-0.28] respectively), but protection dropped after 60 days to 1.04 [1.01-1.06]; 0.52 [0.44-0.61] and 0.38 [0.33-0.45]). Compared with 2-dose-schemes, homologous boosters after primary schemes with vectored-vaccines provided lower protection against infections < and ≥60 days (0.94 [0.92-0.97] and 1.05 [1.01-1.09], respectively) but protected against hospitalisations (0.30 [0.26-0.35]) and deaths (0.29 [0.25-0.33]), decreasing after 60 days (0.59 [0.47-0.74] and 0.51 [0.41-0.64], respectively). Heterologous boosters protected against infections (0.70 [0.68-0.71]) but decreased after 60 days (1.01 [0.98-1.04]) and against hospitalisations and deaths (0.26 [0.22-0.31] and 0.22 [0.18-0.25], respectively), which also decreased after 60 days (0.43 [0.35-0.53] and 0.33 [0.26-0.41], respectively). Heterologous boosters protected against infections when applied <60 days (0.70 [0.68-0.71], p < 0.001), against hospitalisations when applied ≥60 days (0.43 [0.35-0.53], p < 0.01), and against deaths < and ≥60 days (0.22 [0.18-0.25], p < 0.01 and 0.33 [0.26-0.41], p < 0.001). Interpretation: During omicron predominance, heterologous boosters such as viral vectored and mRNA vaccines, following Sputnik V, ChAdOx1, Sinopharm or heterologous primary schemes might provide better protection against death; this effect might last longer in individuals aged ≥50 than homologous boosters. Funding: None.

Efectividad de refuerzos homólogos y heterólogos luego de esquemas primarios con Sputnik V, Astra-Zeneca y Sinopharm durante el período omicron en adultos mayores de 50 años en la provincia de Buenos Aires / Effectiveness of homologous and heterologous boosters after primary regimens with Sputnik V, AstraZeneca, and Sinopharm during the omicron variant period in adults over 50 years old in the province of Buenos Aires

Objetivo: Evaluar la efectividad de esquemas primarios de Sputnik V, Astra-Zeneca, Sinopharm o combinaciones heterólogas seguidos de refuerzos a vector viral (Sputnik V, Astra-Zeneca) o ARNm (Pfizer-BioNTech, Moderna) frente a infecciones, hospitalizaciones y muertes por SARS-CoV-2. Material y métodos: Estudio de casos y controles con test negativo realizado en la provincia de Buenos Aires, durante el predominio de ómicron BA.1, que incluyó individuos ≥ 50 años con test positivo para SARS-CoV-2 que habían recibido 2 o 3 dosis de vacunas. Se registraron infecciones, hospitalizaciones y muertes después de administrar refuerzos con Sputnik V, Astra-Zeneca o ARNm. Resultados: De 422 124 personas analizadas para SARS-Cov-2, 221 993 (52.5%) presentaron test positivos; 190 884 (45.2%) y 231.260 (54.8%) recibieron esquemas de vacunación de 2 y 3 dosis, respectivamente. Los esquemas primarios con Astra-Zeneca, Sputnik V o vector viral, combinados con un refuerzo a vector viral, mostraron protección contra infecciones (OR: 0.94 [0.92 a 0.97]), hospitalizaciones (OR: 0.30 [0.26 a 0.35]) y muertes (OR: 0.29 [0.25 a 0.33]. Los esquemas primarios con Astra-Zeneca y Sputnik V más refuerzo de ARNm, o con Sinopharm más refuerzo de ARNm o vector viral otorgaron protección adicional contra infecciones (OR: 0.70 [0.68 a 0.71]). Hubo un efecto protector frente a hospitalizaciones y muertes (OR: 0.26 [0.22 a 0.31] y 0.22 [0.18 a 0.25]) en todos los casos. Conclusiones: Durante el predominio de ómicron, los refuerzos heterólogos con vacunas a vector viral y de ARNm, posteriores a los esquemas primarios de Sputnik V, Astra-Zeneca, Sinopharm o heterólogos, podrían proporcionar mejor protección y mayor duración del efecto contra la muerte en personas mayores de 50 años, en comparación con refuerzos homólogos

Objective: To evaluate the effectiveness of primary vaccination regimens involving Sputnik V, Astra-Zeneca, Sinopharm, or heterologous combinations followed by viral vector boosters (Sputnik V, As-traZeneca) or mRNA boosters (Pfizer-BioNTech, Moderna) against infections, hospitalizations and deaths caused by SARS-CoV-2. Material and methods: Case-control studies with negative tests conducted in the Buenos Aires province during the Omicron BA.1 predominance. The study included patients ≥ 50 years of age, who tested positive for SARS-CoV-2 and had received two or three doses of vaccines. Infections, hospitalizations, and deaths were registered following the administration of Sputnik V, AstraZeneca, or mRNA boosters. Findings: Out of 422 124 people tested for SARS-Cov-2, 221 993 (52.5%) had positive test results; 190,884 (45.2%) and 231 260 (54.8%) received two-dose and three-dose vaccination schemes, respectively. Primary regimens with AstraZeneca, Sputnik V, or viral vector, combined with a viral vector booster demonstrated protection against infections (OR 0.94 [0.92 to 0.97]), hospitalizations (OR 0.30 [0.26 to 0.35]) and deaths (OR 0.29 [0.25 to 0.33]. Primary regimens with AstraZeneca and Sputnik V combined with mRNA boosters, as well as pri-mary schemes with Sinopharm combined with mRNA or viral vectored boosters showed additional protection against infections (OR 0.70 [0.68 to 0.71]). There was a protective effect against hospi-talizations and deaths (OR 0.26 [0.22-0.31] and 0.22 [0.18 -0.25]) in all cases. Conclusions: During Omicron predominance, heterologous boosters with viral-vector and mRNA vaccines, administered after Sputnik V, AstraZeneca, Sinopharm, or heterologous primary regimens, could provide enhanced protection and prolonged effectiveness against mortality in individuals aged ≥ 50, compared to ho-mologous boosters

Safety and immunogenicity of a SARS-CoV-2 Gamma variant RBD-based protein adjuvanted vaccine used as booster in healthy adults.

A Gamma Variant RBD-based aluminum hydroxide adjuvanted vaccine called ARVAC CG was selected for a first in human clinical trial. Healthy male and female participants (18-55 years old) with a complete COVID-19-primary vaccine scheme were assigned to receive two intramuscular doses of either a low-dose or a high-dose of ARVAC CG. The primary endpoint was safety. The secondary objective was humoral immunogenicity. Cellular immune responses were studied as an exploratory objective. The trial was prospectively registered in PRIISA.BA (Registration Code 6564) and ANMAT and retrospectively registered in ClinicalTrials.gov (NCT05656508). Samples from participants of a surveillance strategy implemented by the Ministry of Health of the Province of Buenos Aires that were boosted with BNT162b2 were also analyzed to compare with the booster effect of ARVAC CG. ARVAC CG exhibits a satisfactory safety profile, a robust and broad booster response of neutralizing antibodies against the Ancestral strain of SARS-CoV-2 and the Gamma, Delta, Omicron BA.1 and Omicron BA.5 variants of concern and a booster effect on T cell immunity in individuals previously immunized with different COVID-19 vaccine platforms.

Utilización precoz de tocilizumab en pacientes internados con COVID-19 grave y crítica: un estudio multicéntrico en la Provincia de Buenos Aires / Early use of tocilizumab in hospitalized patients with severe and critical COVID-19 in the Province of Buenos Aires: a multicentric study

Resumen Introducción: El objetivo principal del estudio fue evaluar la mortalidad en los pacientes con COVID-19 graves y críticos, que recibieron tocilizumab (TCZ) -un antagonista monoclonal del receptor de IL-6- de forma temprana vs. tardía. Métodos: Cohorte retrospectiva multicéntrica de pacientes >18 años internados con COVID-19 desde el 1/7/2021-1/8/2022, con 5-7 puntos de gravedad inicial (GI) según Escala de la OMS. Se consideró adminis tración temprana o tardía a la infusión de TCZ ≤ ó > a 48 h del ingreso. Las variables de resultado fueron mortalidad a 28 días y cambio de la GI. Los factores relacionados con la mortalidad fueron evaluados con regresión de Cox. Resultados: Se incluyeron 266 pacientes, 159(60%) varones; edad 58(± 15); con hipertensión arterial (43%), obesidad (37%) y diabetes (27%);70 presentaban GI = 5 (oxígeno suplementario), 143 GI = 6 (ventilación no inva siva o cánula nasal de alto flujo) y 53 GI = 7 (ventilación mecánica invasiva). La mortalidad a 28 días fue 42%, asociada independientemente a: edad, obesidad, GI, días entre la internación y administración del TCZ, y días entre la fecha de inicio de síntomas y el TCZ. La mortalidad para GI 5, 6 y 7 fue 26%, 39% y 72%, respectivamente; 76% y 62% de los pacientes permanecieron estables o mejoraron la GI a los días 3 y 7 de la infusión de TCZ. La mortalidad a 28 días fue 39% (TCZ temprano) vs. 57% (TCZ tardío); p = 0.02; HR = 0.63[0.41-0.99, p = 0.05]). Discusión: Estos resultados apoyan la administración temprana de TCZ en pacientes con COVID-19 grave y crítica.

Abstract Introduction: Tocilizumab (TCZ), an IL-6 receptor antagonist monoclonal antibody is warranted in severe and critically-ill COVID-19 patients. The objective was to evaluate 28-day mortality of patients with severe or critical COVID-19 treated with early vs delayed TCZ. Methods: Multicenter, retrospective cohort study in cluding patients>18 years hospitalized between 7/1/2021- 8/1/2022 with confirmed COVID-19, with 5, 6 and 7 points of WHO Ordinal Initial Severity Scale [SS]. Early or late administration was considered if TCZ was administered before or after 48 hours from admission. Outcomes were28-day mortality and change of SS. Factors related to 28-day mortality were evaluated with Cox regression. Results: 266 patients were included, 159(60%) male; aged 58(± 15); frequent comorbidities were hypertension (42%), obesity (37%) and diabetes (27%). Seventy patients had a SS = 5 (Supplemental O2), 143 had SS = 6 (NIV/ HFNC), and 53 had SS = 7 (IMV). 28-day mortality was 42%(112/266); predictors were age, obesity, higher SS, days between hospitalization and TCZ administration, and fewer days between symptoms onset and TCZ. Mortality of SS 5, 6 and 7 was 26%, 39% and 72% respectively. Com pared with baseline SS points, 76% and 62% of patients remained stable or improved on days 3 and 7 since TCZ administration. 28-day mortality was lower when TCZ was administered before 48 hours (39% vs 57%; p = 0.02; HR = 0.63;[0.41-0.99, p = 0.05]). Discussion: This study supports the early use of TCZ in patients with severe or critical COVID-19.

[Early use of tocilizumab in hospitalized patients with severe and critical COVID-19 in the Province of Buenos Aires: a multicentric study]. / Utilización precoz de tocilizumab en pacientes internados con COVID-19 grave y crítica: un estudio multicéntrico en la Provincia de Buenos Aires.

INTRODUCTION: Tocilizumab (TCZ), an IL-6 receptor antagonist monoclonal antibody is warranted in severe and critically-ill COVID-19 patients. The objective was to evaluate 28-day mortality of patients with severe or critical COVID-19 treated with early vs delayed TCZ. METHODS: Multicenter, retrospective cohort study including patients >18 years hospitalized between 7/1/2021-8/1/2022 with confirmed COVID-19, with 5, 6 and 7 points of WHO Ordinal Initial Severity Scale [SS]. Early or late administration was considered if TCZ was administered before or after 48 hours from admission. Outcomes were 28-day mortality and change of SS. Factors related to 28-day mortality were evaluated with Cox regression. RESULTS: 266 patients were included, 159(60%) male; aged 58(± 15); frequent comorbidities were hypertension (42%), obesity (37%) and diabetes (27%). Seventy patients had a SS = 5 (Supplemental O2), 143 had SS = 6 (NIV/ HFNC), and 53 had SS = 7 (IMV). 28-day mortality was 42%(112/266); predictors were age, obesity, higher SS, days between hospitalization and TCZ administration, and fewer days between symptoms onset and TCZ. Mortality of SS 5, 6 and 7 was 26%, 39% and 72% respectively. Compared with baseline SS points, 76% and 62% of patients remained stable or improved on days 3 and 7 since TCZ administration. 28-day mortality was lower when TCZ was administered before 48 hours (39% vs 57%; p = 0.02; HR = 0.63;[0.41-0.99, p = 0.05]). DISCUSSION: This study supports the early use of TCZ in patients with severe or critical COVID-19.

Introducción: El objetivo principal del estudio fue evaluar la mortalidad en los pacientes con COVID-19 graves y críticos, que recibieron tocilizumab (TCZ) -un antagonista monoclonal del receptor de IL-6- de forma temprana vs. tardía. Métodos: Cohorte retrospectiva multicéntrica de pacientes > 18 años internados con COVID-19 desde el 1/7/2021-1/8/2022, con 5-7 puntos de gravedad inicial (GI) según Escala de la OMS. Se consideró administración temprana o tardía a la infusión de TCZ = ó > a 48 h del ingreso. Las variables de resultado fueron mortalidad a 28 días y cambio de la GI. Los factores relacionados con la mortalidad fueron evaluados con regresión de Cox. Resultados: Se incluyeron 266 pacientes, 159(60%) varones; edad 58(± 15); con hipertensión arterial (43%), obesidad (37%) y diabetes (27%);70 presentaban GI = 5 (oxígeno suplementario), 143 GI = 6 (ventilación no invasiva o cánula nasal de alto flujo) y 53 GI = 7 (ventilación mecánica invasiva). La mortalidad a 28 días fue 42%, asociada independientemente a: edad, obesidad, GI, días entre la internación y administración del TCZ, y días entre la fecha de inicio de síntomas y el TCZ. La mortalidad para GI 5, 6 y 7 fue 26%, 39% y 72%, respectivamente; 76% y 62% de los pacientes permanecieron estables o mejoraron la GI a los días 3 y 7 de la infusión de TCZ. La mortalidad a 28 días fue 39% (TCZ temprano) vs. 57% (TCZ tardío); p = 0.02; HR = 0.63[0.41-0.99, p = 0.05]). Discusión: Estos resultados apoyan la administración temprana de TCZ en pacientes con COVID-19 grave y crítica.

Effectiveness of BBIBP-CorV, BNT162b2 and mRNA-1273 vaccines against hospitalisations among children and adolescents during the Omicron outbreak in Argentina: A retrospective cohort study.

Background: Although paediatric clinical presentations of COVID-19 are usually less severe than in adults, serious illness and death have occurred. Many countries started the vaccination rollout of children in 2021; still, information about effectiveness in the real-world setting is scarce. The aim of our study was to evaluate vaccine effectiveness (VE) against COVID-19-associated-hospitalisations in the 3-17-year population during the Omicron outbreak. Methods: We conducted a retrospective cohort study including individuals aged 3-17 registered in the online vaccination system of the Buenos Aires Province, Argentina. mRNA-1273 and BNT162b2 were administered to 12-17-year subjects; and BBIBP-CorV to 3-11-year subjects. Vaccinated group had received a two-dose scheme by 12/1/2021. Unvaccinated group did not receive any COVID-19 vaccine between 12/14/2021 and 3/9/2022, which was the entire monitoring period. Vaccine effectiveness (VE) against COVID-19-associated hospitalisations was calculated as (1-OR)x100. Findings: By 12/1/2021, 1,536,435 individuals aged 3-17 who had received zero or two doses of SARS-CoV-2 vaccines were included in this study. Of the latter, 1,440,389 were vaccinated and 96,046 not vaccinated. VE were 78.0%[68.7-84.2], 76.4%[62.9-84.5] and 80.0%[64.3-88.0] for the entire cohort, 3-11-year (BBIBP-CorV) subgroup and 12-17 (mRNA vaccines) subgroup, respectively. VE for the entire population was 82.7% during the period of Delta and Omicron overlapping circulation and decreased to 67.7% when Omicron was the only variant present. Interpretation: This report provides evidence of high vaccine protection against associated hospitalisations in the paediatric population during the Omicron outbreak but suggests a decrease of protection when Omicron became predominant. Application of a booster dose in children aged 3-11-year warrants further consideration. Funding: None.

Timing of convalescent plasma administration and 28-day mortality in COVID-19 pneumonia.

This is a multicenter cohort study including consecutive, hospitalized patients ≥18 years, with moderate to severe COVID-19, carried out to evaluate the relationship between the timing of convalescent plasma administration and 28-day mortality. Data were prospectively collected between May 14, 2020 and October 31, 2020. Patients were grouped according to the timing of administration of convalescent plasma as <3 days, between 3 and 7 days, and >7 days. The main outcome variable was 28-day mortality. Independent predictors of mortality were identified by logistic regression. Of 4719 patients receiving convalescent plasma, 3036 (64.3%) were in the general ward, 1171 (24.8%) in the intensive care unit (ICU), and 512 (10.8%) in the ICU on mechanical ventilation. Convalescent plasma was administered to 3113 (66%) patients within the first 3 days of hospital admission, to 1380 (29.2%) between 3 and 7 days, and to 226 after 7 days; 28-day mortality was, respectively, 18.1%, 30.4% and 38.9% (p<0.001). In the regression model, convalescent plasma administration within the first 3 days of admission was associated with reduced 28-day mortality, compared with the administration after 7 days (OR 0.40, 95% CI 0.30 to 0.53). Early convalescent plasma administration was associated to a significant decreased mortality in patients in the general ward (OR 0.45, 95% CI 0.29 to 0.69) and in the ICU (OR 0.35, 95% CI 0.19 to 0.64), but not in those requiring mechanical ventilation (OR 0.52, 95% CI 0.27 to 1.01). In conclusion, this study suggests that early administration of convalescent plasma to patients with COVID-19 pneumonia is critical to obtain therapeutic benefit.

Cysteine-Rich Secretory Proteins (CRISP) are Key Players in Mammalian Fertilization and Fertility.

Mammalian fertilization is a complex process involving a series of successive sperm-egg interaction steps mediated by different molecules and mechanisms. Studies carried out during the past 30 years, using a group of proteins named CRISP (Cysteine-RIch Secretory Proteins), have significantly contributed to elucidating the molecular mechanisms underlying mammalian gamete interaction. The CRISP family is composed of four members (i.e., CRISP1-4) in mammals, mainly expressed in the male tract, present in spermatozoa and exhibiting Ca2+ channel regulatory abilities. Biochemical, molecular and genetic approaches show that each CRISP protein participates in more than one stage of gamete interaction (i.e., cumulus penetration, sperm-ZP binding, ZP penetration, gamete fusion) by either ligand-receptor interactions or the regulation of several capacitation-associated events (i.e., protein tyrosine phosphorylation, acrosome reaction, hyperactivation, etc.) likely through their ability to regulate different sperm ion channels. Moreover, deletion of different numbers and combination of Crisp genes leading to the generation of single, double, triple and quadruple knockout mice showed that CRISP proteins are essential for male fertility and are involved not only in gamete interaction but also in previous and subsequent steps such as sperm transport within the female tract and early embryo development. Collectively, these observations reveal that CRISP have evolved to perform redundant as well as specialized functions and are organized in functional modules within the family that work through independent pathways and contribute distinctly to fertility success. Redundancy and compensation mechanisms within protein families are particularly important for spermatozoa which are transcriptionally and translationally inactive cells carrying numerous protein families, emphasizing the importance of generating multiple knockout models to unmask the true functional relevance of family proteins. Considering the high sequence and functional homology between rodent and human CRISP proteins, these observations will contribute to a better understanding and diagnosis of human infertility as well as the development of new contraceptive options.

Effectiveness of the first component of Gam-COVID-Vac (Sputnik V) on reduction of SARS-CoV-2 confirmed infections, hospitalisations and mortality in patients aged 60-79: a retrospective cohort study in Argentina.

BACKGROUND: A first-dose of various vaccines provides acceptable protection against infections by SARS-CoV-2 and evolution to the most severe forms of COVID-19. The recombinant adenovirus (rAd)-based vaccine, Gam-COVID-Vac (Sputnik V), was proven efficacious but information about effectiveness in the real-world setting is lacking. The aim of our study was to investigate the association between the rollout of the first component (rAd26) of Gam-COVID-Vac and PCR-positive tests, hospitalisations and deaths. METHODS: We conducted a retrospective cohort study which analyzed individuals aged 60-79 who self-registered in the online vaccination system of the Province of Buenos Aires, Argentina, from December 29, 2020 to March 21, 2021. Exclusion criteria were having a previous positive RT-PCR or antigen tests for SARS-CoV-2, having received other vaccines, or two doses of any vaccine.Proportions of new laboratory-confirmed SARS-CoV-2 infections, hospitalisations and deaths until 83 days of vaccination were compared between vaccinated and unvaccinated subjects. Vaccine effectiveness for the three outcomes was calculated as (1-OR) × 100. Kaplan-Meier cumulative incidence curves were constructed. FINDINGS: During the study period 415995 registered subjects received the first component of Gam-COVID-Vac; 40387 belonged to the 60-79 age group, and were compared to 38978 unvaccinated. Vaccine effectiveness for preventing laboratory-confirmed infections was 78•6% [CI95% 74·8 - 81·7]; and for reducing hospitalizations and deaths was, respectively, 87·6% [CI95% 80·3 - 92·2] and 84·8% [CI95% 75·0 - 90·7]. Effectiveness was high across all subgroups. INTERPRETATION: Similarly to other vaccines, the administration of one dose of Gam-COVID-Vac was effective for a wide range of COVID-19-related outcomes. FUNDING: This study did not receive any funding.

Pharmacological Inactivation of CatSper Blocks Sperm Fertilizing Ability Independently of the Capacitation Status of the Cells: Implications for Non-hormonal Contraception.

Cation channel of sperm (CatSper), the main sperm-specific Ca2+ channel, plays a key role in mammalian fertilization, and it is essential for male fertility, becoming an attractive target for contraception. Based on this, in the present work, we investigated the effects of CatSper inactivation on in vitro and in vivo sperm fertilizing ability and the mechanisms underlying such effects. Exposure of cauda epididymal mouse sperm to different concentrations (1-20 µM) of the potent CatSper inhibitor HC-056456 (HC) during in vitro capacitation showed no effects on sperm viability but significantly affected Ca2+ entry into the cells, progressive motility, protein tyrosine phosphorylation, induced acrosome reaction, and hyperactivation, as well as the sperm's ability to in vitro fertilize cumulus oocyte complexes and zona-free eggs. Whereas the presence of HC during gamete coincubation did not affect in vitro fertilization, exposure of either non-capacitating or already capacitated sperm to HC prior to gamete coincubation severely reduced fertilization, indicating that sperm function is affected by HC when the cells are incubated with the drug before sperm-egg interaction. Of note, insemination of HC-treated sperm into the uterus significantly or completely reduced the percentage of oviductal fertilized eggs showing, for the first time, the effects of a CatSper inhibitor on in vivo fertilization. These observations, together with the finding that HC affects sperm fertilizing ability independently of the sperm capacitation status, provide further insights on how CatSper regulates sperm function and represent a solid proof of concept for developing a male/female non-hormonal contraceptive based on the pharmacological blockage of CatSper activity.

Risk factors for COVID-19 mortality: The effect of convalescent plasma administration.

BACKGROUND: Convalescent plasma, widely utilized in viral infections that induce neutralizing antibodies, has been proposed for COVID-19, and preliminary evidence shows that it might have beneficial effect. Our objective was to determine the risk factors for 28-days mortality in patients who received convalescent plasma for COVID-19 compared to those who did not, who were admitted to hospitals in Buenos Aires Province, Argentina, throughout the pandemic. METHODS: This is a multicenter, retrospective cohort study of 2-month duration beginning on June 1, 2020, including unselected, consecutive adult patients with diagnosed COVID-19, admitted to 215 hospitals with pneumonia. Epidemiological and clinical variables were registered in the Provincial Hospital Bed Management System. Convalescent plasma was supplied as part of a centralized, expanded access program. RESULTS: We analyzed 3,529 patients with pneumonia, predominantly male, aged 62±17, with arterial hypertension and diabetes as main comorbidities; 51.4% were admitted to the ward, 27.1% to the Intensive Care Unit (ICU), and 21.7% to the ICU with mechanical ventilation requirement (ICU-MV). 28-day mortality was 34.9%; and was 26.3%, 30.1% and 61.4% for ward, ICU and ICU-MV patients. Convalescent plasma was administered to 868 patients (24.6%); their 28-day mortality was significantly lower (25.5% vs. 38.0%, p<0.001). No major adverse effects occurred. Logistic regression analysis identified age, ICU admission with and without MV requirement, diabetes, and preexistent cardiovascular disease as independent predictors of 28-day mortality, whereas convalescent plasma administration acted as a protective factor. CONCLUSIONS: Our study suggests that the administration of convalescent plasma in COVID-19 pneumonia admitted to the hospital might be associated with improved outcomes.

Impact of vaccines against COVID-19 on new SARS-COV2 infections in health care workers of the Province of Buenos Aires / Impacto de vacunación covid-19 en las infecciones por sars-cov-2 en personal de salud de la provincia de buenos aires

RESUMEN INTRODUCCIÓN : La vacunación en la provincia de Buenos Aires inició con personal de salud (PS). El objetivo de este trabajo es evaluar el impacto de la vacunación sobre la evolución de los casos de infección por SARS-CoV-2 en el PS, comparado con la de la población general (PG). MÉTODOS : Estudio obsewacional descriptivo de series temporales de casos confirmados de COVID-19, entre abril de 2020 y marzo de 2021. Se utilizó el sistema de información VacunatePBA y el SNVS. Se tomó el grado de inmunización a 14 días de la aplicación de la vacuna. RESULTADOS : A partir de septiembre de 2020, el número de casos disminuyó en el PS, en comparación con la PG. Para fines de febrero de 2021 estaba inmunizado con primera dosis (PD) el 42% del PS y con segunda dosis (SD) el 24%, mientras que en PG solo estaba inmunizado el 0,06% con PD y nadie SD. En marzo de 2021 mientras la cantidad de casos en PG (con 2% inmunizada) aumentó un 10% con respecto a febrero, en PS disminuyó un 35% (95% de esa población inmunizada). Esto indica una disminución significativa entre los nuevos casos de febrero y de marzo en PS (p <0,00001). DISCUSIÓN : A partir del inicio de la vacunación contra COVID-19 a finales de diciembre 2020, se muestra una disminución de casos nuevos de COVID-19 en PS. Este trabajo describe una disminución relativa de los casos en PS luego de la vacunación en la provincia de Buenos Aires y aporta los primeros datos del país sobre el impacto de las vacunas contra COVID-19.

ABSTRACT INTRODUCTION : The vaccination in the province of Buenos Aires has initiated with the health care workers (HCW). The present work aims to evaluate the impact of vaccination on the evolution of confirmed SARS- CoV2 cases in HCW compared to that of the general population (GP). METHODS : The study design is descriptive observational of series were developed confirmed cases, period April 2020 to March 2021. The Sistema Nacional de Vigilancia en Salud (SNVS), and VacunatePBA were used. The grade immunization was estimated at 14 days from the application of the first dose. RESULTS : As of September, the number of cases begins to decrease more sharply in HCW than in the general population. By February 2021, 42% of the HCW had been immunized with the first dose (FD) and 24% with the second dose (SD), while in GP only 0.06% had been immunized with FD and 0% of SD. In March, while the number of GP cases increased by 10% compared to February (only 2% immunized), HCW decreased by 35% (95% immunized). This indicates a significant decrease between the new cases of February and March in HCW (p-value <0.00001). DISCUSSION: Since the introduction of COVID-19 vaccines at the end of December in different countries, a decrease in new cases of COVID-19 in HCW has begun to be reported. The present work describes a relative decrease in HCW cases in a post-vaccination context in the province of Buenos Aires, providing the first data in the country on the impact of COVID-19 vaccines on HCW.

Recurrent Somatic Chromosomal Abnormalities in Relapsed Extraocular Retinoblastoma.

Most reports about copy number alterations (CNA) in retinoblastoma relate to patients with intraocular disease and features of children with extraocular relapse remain unknown, so we aimed to describe the CNA in this population. We evaluated 23 patients and 27 specimens from 4 centers. Seventeen cases had extraocular relapse after initial enucleation and six cases after an initial preservation attempt. We performed an analysis of CNA and BCOR gene alteration by SNP array (Single Nucleotide Polymorfism array), whole-exome sequencing, IMPACT panel and CGH array (Array-based comparative genomic hybridization). All cases presented CNA at a higher prevalence than those reported in previously published studies for intraocular cases. CNA previously reported for intraocular retinoblastoma were found at a high frequency in our cohort: gains in 1q (69.5%), 2p (60.9%) and 6p (86.9%), and 16q loss (78.2%). Other, previously less-recognized, CNA were found including loss of 11q (34.8%), gain of 17q (56.5%), loss of 19q (30.4%) and BCOR alterations were present in 72.7% of our cases. A high number of CNA including 11q deletions, 17q gains, 19q loss, and BCOR alterations, are more common in extraocular retinoblastoma. Identification of these features may be correlated with a more aggressive tumor warranting consideration for patient management.

Design, synthesis, and evaluation of substrate - analogue inhibitors of Trypanosoma cruzi ribose 5-phosphate isomerase type B.

Ribose 5-phosphate isomerase type B (RPI-B) is a key enzyme of the pentose phosphate pathway that catalyzes the isomerization of ribose 5-phosphate (R5P) and ribulose 5-phosphate (Ru5P). Trypanosoma cruzi RPI-B (TcRPI-B) appears to be a suitable drug-target mainly due to: (i) its essentiality (as previously shown in other trypanosomatids), (ii) it does not present a homologue in mammalian genomes sequenced thus far, and (iii) it participates in the production of NADPH and nucleotide/nucleic acid synthesis that are critical for parasite cell survival. In this survey, we report on the competitive inhibition of TcRPI-B by a substrate - analogue inhibitor, Compound B (Ki = 5.5 ± 0.1 µM), by the Dixon method. This compound has an iodoacetamide moiety that is susceptible to nucleophilic attack, particularly by the cysteine thiol group. Compound B was conceived to specifically target Cys-69, an important active site residue. By incubating TcRPI-B with Compound B, a trypsin digestion LC-MS/MS analysis revealed the identification of Compound B covalently bound to Cys-69. This inhibitor also exhibited notable in vitro trypanocidal activity against T. cruzi infective life-stages co-cultured in NIH-3T3 murine host cells (IC50 = 17.40 ± 1.055 µM). The study of Compound B served as a proof-of-concept so that next generation inhibitors can potentially be developed with a focus on using a prodrug group in replacement of the iodoacetamide moiety, thus representing an attractive starting point for the future treatment of Chagas' disease.

Impacto de vacunación de COVID-19 en las infecciones por SARS-COV-2 en personal de salud de la provincia de Buenos Aires / Impact of vaccines against COVID-19 on new SARS-COV2 infections in health care workers of the Province of Buenos Aires

INTRODUCCIÓN: La vacunación en la provincia de Buenos Aires inició con personal de salud (PS). El objetivo de este trabajo es evaluar el impacto de la vacunación sobre la evolución de los casos de infección por SARS-CoV-2 en el PS, comparado con la de la población general (PG). MÉTODOS: Estudio observacional descriptivo de series temporales de casos confirmados de COVID-19, entre abril de 2020 y marzo de 2021. Se utilizó el sistema de información VacunatePBA y el SNVS. Se tomó el grado de inmunización a 14 días de la aplicación de la vacuna. RESULTADOS: A partir de septiembre de 2020, el número de casos disminuyó en el PS, en comparación con la PG. Para fines de febrero de 2021 estaba inmunizado con primera dosis (PD) el 42% del PS y con segunda dosis (SD) el 24%, mientras que en PG solo estaba inmunizado el 0,06% con PD y nadie SD. En marzo de 2021 mientras la cantidad de casos en PG (con 2% inmunizada) aumentó un 10% con respecto a febrero, en PS disminuyó un 35% (95% de esa población inmunizada). Esto indica una disminución significativa entre los nuevos casos de febrero y de marzo en PS (p <0,00001). DISCUSIÓN: A partir del inicio de la vacunación contra COVID-19 a finales de diciembre 2020, se muestra una disminución de casos nuevos de COVID-19 en PS. Este trabajo describe una disminución relativa de los casos en PS luego de la vacunación en la provincia de Buenos Aires y aporta los primeros datos del país sobre el impacto de las vacunas contra COVID-19

Compromiso cardiovascular en pacientes con síndrome inflamatorio pediátrico multisistémico, asociado a infección por SARS-CoV-2 / Cardiovascular involvement in pediatric patients with SARS-CoV-2 associated to multisystemic inflammatory syndrome

INTRODUCCION: El Síndrome Inflamatorio Pediátrico Multisistémico (PIMS) ha emergido como una nueva enfermedad en niños, secundaria a infección por SARSCoV-2. Se caracteriza por presentar compromiso multiorgánico con parámetros inflamatorios elevados y manifestaciones clínicas graves, siendo el corazón el órgano más severamente comprometido. OBJETIVO: Describir las características clínicas y de laboratorio de 23 pacientes con diagnóstico de PIMS con compromiso cardiovascular hospitalizados en un centro único. MÉTODO: Se efectuó un estudio retrospectivo analizando los hallazgos clínicos y de laboratorio junto a las manifestaciones cardiovasculares que presentaron estos pacientes. RESULTADOS: 23/29 pacientes con PIMS (78%) presentaron manifestaciones digestivas y mucocutáneas. Las manifestaciones cardiovasculares fueron: Síndrome Kawasaki y "Kawasaki like" sin compromiso coronario en 15/23 (65%) y con compromiso coronario en 3 (13%). Shock en 9 pacientes (39%), injuria miocárdica- miocarditis en 8 (35%) y derrame pericárdico en 13 (56%). Trastornos del ritmo cardíaco se observaron en 6 pacientes (26%). La terapia más utilizada fue inmunoglobulina y corticoides. 18 /23 requirieron manejo en unidades de intermedio y/o intensivo. Un 70% de los pacientes se recuperó del compromiso cardiovascular antes del alta. CONCLUSIÓN: El compromiso cardiovascular en PIMS es la complicación más frecuente de esta enfermedad, que se acompaña de manifestaciones inmunológicas y hematológicas graves lo que hace necesario un tratamiento multidisciplinario para un mejor manejo de estos pacientes.

INTRODUCTION: Pediatric Multisystemic Inflammatory Syndrome (PIMS) has emerged as a new disease in children, secondary to SARSCoV-2 infection. It is characterized by multi-organ involvement with elevated inflammatory parameters and severe clinical manifestations, the heart being the organ most severely involved. OBJETIVE: to describe the clinical and laboratory characteristics of 23 patients diagnosed with PIMS with cardiovascular involvement hospitalized in a single center. METHOD: We conducted a retrospective study in which we analyzed the clinical and laboratory findings along with the cardiovascular manifestations presented by these patients. Results: 23/29 patients with PIMS and cardiovascular involvement were selected, 78% had digestive and mucocutaneous manifestations. Cardiovascular manifestations consisted of KawasakiKawasaki like syndrome without coronary involvement in 15/23 (65%) and coronary involvement in 3 (13%). Nine patients developed shock (39%), 8 (35%) myocardial injury in and 13 (56%) pericardial effusion.. Heart rhythm disorders were observed in 6 patients (26%). The main therapy was immunoglobulin and corticosteroids. 18 /23 required management in intermediate and/or intensive care unit. 70% of patients recovered from cardiovascular involvement before discharge. CONCLUSION: Cardiovascular involvement in PIMS is the most frequent complication of this disease, but it is associated with severe immunological and hematological manifestations, which makes necessary a multidisciplinary treatment for a better management

[Convalescent plasma therapy in COVID-19 patients, in the Province of Buenos Aires]. / Terapia con plasma de convalecientes en pacientes COVID-19 en la Provincia de Buenos Aires.

This is a preliminary, multicenter, retrospective cohort study, including 272 consecutive patients with COVID-19 admitted to hospitals in Buenos Aires Province, between May 15th and July 1st, 2020, included in an expanded access program to convalescent plasma. Our objectives were to analyze mortality and its independent risk factors, and to assess the occurrence of a favorable evolution, defined as hospital discharge, or stay at the ward, or transfer from ICU to ward. Patients were stratified int o 4 subgroups: admission to the ward with pneumonia and/or oxygen requirement (WARD; n = 100); ICU admission (ICU; n = 87); ICU admission with requirement of mechanical ventilation (ICU-MV; n = 56), and ICU-MV plus septic shock (ICU-MV-SS; N = 29). Mortality at 28 days was 26.1% for the entire group, 14.0% for WARD group, 18.4% for ICU, 44.6% for ICU-MV, and 55.2% for ICU-MV-SS. Mean survival time (days) was 25.6 ± 0.6 (WARD); 25.3 ± 0.7 (ICU); 20.8 ± 1.2 (ICU-MV) and 18.2 ± 1.8 (ICU-MV-SS). Independent predictors of mortality were MV, septic shock and weight. A favorable evolution occurred in 81.4% of WARD patients; in 70.9% of ICU; in 39.6% of ICU-MV and in 27.6% of ICU-MV-SS patients. Severity of illness on admission, age, weight and heart rate were independently associated with evolution. No major adverse effects were recorded. The lack of a control group precluded the estimation of efficacy. However, our 26% mortality rate was higher than that of the treatment arm of clinical trials comparing plasma with usual treatment, which might be ascribed to higher proportion of patients with MV and septic shock in our cohort.

Se trata de un estudio multicéntrico de cohorte retrospectivo, observacional, desde 15/5 a 1/7, 2020, en 272 pacientes COVID-19 internados en hospitales de la provincia de Buenos Aires, incluidos en un programa de acceso expandido de plasma de convalecientes de COVID-19. Nuestros objetivos fueron analizar letalidad y sus factores de riesgo independientes, y evaluar la evolución favorable, definida como alta hospitalaria, permanencia en sala (PISO), o alta de la UTI. Los pacientes fueron estratificados en 4 subgrupos: ingreso a PISO (n = 100) con neumonía y/o requerimiento de oxígeno; a UTI (n = 87); a UTI con requerimiento de ventilación mecánica (UTI-VM; n = 56), y a UTI-VM con shock séptico (UTI-VM-SS; n = 29). La letalidad total a los 28 días fue 26.1%, (71/272), para PISO 14.0%; UTI, 18.4%; UTI-VM, 44.6%; y UTI-VM-SS, 55.2%. El tiempo medio de supervivencia (días): 25.6 ± 0.6 (PISO); 25.3 ± 0.7 (UTI); 20.8 ± 1.2 (UTI-VM) y 18.2 ± 1.8 (UTI- VMSS). Los predictores independientes de letalidad fueron VM, shock séptico y peso. Se registró una evolución favorable en 81.4% de los pacientes en PISO; 70.9% en UTI, 39.6% en UTI-VM, y en 27.6% de UTI-VM-SS. La gravedad al ingreso, edad, peso y frecuencia cardíaca fueron predictores independientes de evolución. No se registraron efectos adversos graves. Por falta de un grupo control, no fue posible evaluar la eficacia del plasma de convaleciente. La letalidad (26%) fue mayor que en otros ensayos clínicos con plasma convaleciente; esto podría deberse a mayor proporción de aquellos con VM y shock séptico en nuestra cohorte.

Terapia con plasma de convalecientes en pacientes Covid-19 en la Provincia de Buenos Aires / Convalescent plasma therapy in COVID-19 patients, in the Province of Buenos Aires

Resumen Se trata de un estudio multicéntrico de cohorte retrospectivo, observacional, desde 15/5 a 1/7, 2020, en 272 pacientes COVID-19 internados en hospitales de la provincia de Buenos Aires, incluidos en un programa de acceso expandido de plasma de convalecientes de COVID-19. Nuestros objetivos fueron analizar letalidad y sus factores de riesgo independientes, y evaluar la evolución favorable, definida como alta hospitalaria, permanencia en sala (PISO), o alta de la UTI. Los pacientes fueron estratificados en 4 subgrupos: ingreso a PISO (n = 100) con neumonía y/o requerimiento de oxígeno; a UTI (n = 87); a UTI con requerimiento de ventilación mecánica (UTI-VM; n = 56), y a UTI-VM con shock séptico (UTI-VM-SS; n = 29). La letalidad total a los 28 días fue 26.1%, (71/272), para PISO 14.0%; UTI, 18.4%; UTI-VM, 44.6%; y UTI-VM-SS, 55.2%. El tiempo medio de supervivencia (días): 25.6 ± 0.6 (PISO); 25.3 ± 0.7 (UTI); 20.8 ± 1.2 (UTI-VM) y 18.2 ± 1.8 (UTIVMSS). Los predictores independientes de letalidad fueron VM, shock séptico y peso. Se registró una evolución favorable en 81.4% de los pacientes en PISO; 70.9% en UTI, 39.6% en UTI-VM, y en 27.6% de UTI-VM-SS. La gravedad al ingreso, edad, peso y frecuencia cardíaca fueron predictores independientes de evolución. No se registraron efectos adversos graves. Por falta de un grupo control, no fue posible evaluar la eficacia del plasma de convaleciente. La letalidad (26%) fue mayor que en otros ensayos clínicos con plasma convaleciente; esto podría deberse a mayor proporción de aquellos con VM y shock séptico en nuestra cohorte.

Abstract This is a preliminary, multicenter, retrospective cohort study, including 272 consecutive patients with COVID-19 admitted to hospitals in Buenos Aires Province, between May 15th and July 1st, 2020, included in an expanded access program to convalescent plasma. Our objectives were to analyze mortality and its independent risk factors, and to assess the occurrence of a favorable evolution, defined as hospital discharge, or stay at the ward, or transfer from ICU to ward. Patients were stratified into 4 subgroups: admission to the ward with pneumonia and/or oxygen requirement (WARD; n = 100); ICU admission (ICU; n = 87); ICU admission with requirement of mechanical ventilation (ICU-MV; n = 56), and ICU-MV plus septic shock (ICU-MV-SS; N = 29). Mortality at 28 days was 26.1% for the entire group, 14.0% for WARD group, 18.4% for ICU, 44.6% for ICU-MV, and 55.2% for ICU-MV-SS. Mean survival time (days) was 25.6±0.6 (WARD); 25.3±0.7 (ICU); 20.8±1.2 (ICU-MV) and 18.2 ± 1.8 (ICU-MV-SS). Independent predictors of mortality were MV, septic shock and weight. A favorable evolution occurred in 81.4% of WARD patients; in 70.9% of ICU; in 39.6% of ICU-MV and in 27.6% of ICU-MV-SS patients. Severity of illness on admission, age, weight and heart rate were independently associated with evolution. No major adverse effects were recorded. The lack of a control group precluded the estimation of efficacy. However, our 26% mortality rate was higher than that of the treatment arm of clinical trials comparing plasma with usual treatment, which might be ascribed to higher proportion of patients with MV and septic shock in our cohort.