Real-Time Imaging of Calcium Dynamics in Human Sperm After Precise Single-Cell Stimulation.

Calcium signaling is a critical regulator of sperm activation and function during the processes of capacitation and fertilization. Here, we describe a combined method for calcium imaging of single, live human sperm in response to stimuli administered with a precisely targeted delivery technique. This protocol is an adaptation of techniques developed for studies of murine sperm [1, 2], and enables real-time monitoring of human sperm calcium dynamics with high spatiotemporal resolution and concurrent detection of acrosome exocytosis (AE), a functional endpoint of sperm capacitation and requirement for physiological fertilization.

Real-Time Imaging of Acrosomal Calcium Dynamics and Exocytosis in Live Mouse Sperm.

Acrosome exocytosis (AE), in which the sperm's single exocytotic vesicle fuses with the plasma membrane, is a complex, calcium-dependent process essential for fertilization. However, our understanding of how calcium signaling regulates AE is still incomplete. In particular, the interplay between intra-acrosomal calcium dynamics and the intermediate steps leading to AE is not well-defined.

Facilitated Asynchronous Online Learning to Build Public Health Strategic Skills.

Context: The COVID-19 pandemic spurred significant government investments for hiring public health workers. There are clear opportunities to help build capacities among both current and incoming public health workers, closing well-elucidated skill gaps.

Objective: To report on the development process, methods used, and outcomes seen from a point-in-time public health workforce capacity-building initiative, Public Health Essentials (PHE) .

Defects in sperm capacitation/fertilizing ability are equally prevalent across ages in men seeking fertility assistance.

Research Question: How do capacitation ability, measured by Cap-Score™, and traditional semen analysis measures (volume, concentration, motility) change with age in men questioning their fertility (MQF)?

Design: Cap-Score and semen analysis measures were obtained from MQF (n = 2652; multicentric design: 35 reproductive endocrinologist prescribers, n = 16 clinics). Morphology was not included due to differences among clinics. A Mann-Whitney test was used to compare Cap-Scores between MQF and men with known recent paternity (n = 76).

Integrated Surveillance System for Controlling COVID-19 on a University Campus, 2020‒2021.

To minimize the impacts of COVID-19 and to keep campus open, Cornell University's Ithaca, NY, campus implemented a comprehensive process to monitor COVID-19 spread, support prevention practices, and assess early warning indicators linked to knowledge, behaviors, and attitudes of campus community members. The integrated surveillance approach informed leadership and allowed for prompt adjustments to university policies and practices through evidence-based decisions. This approach enhanced healthy behaviors and promoted the well-being and safety of all community members.

Case-control study to identify risk factors for SARS-CoV-2 infection among university students in the northeastern USA.

Curbing the coronavirus disease 2019 (COVID-19) pandemic requires a thorough understanding of risk factors for transmission of SARS-CoV-2, the etiologic agent. Institutions of higher education present unique challenges for controlling disease spread because of features inherent to these settings. Our objective was to determine risk factors for SARS-CoV-2 infection among a university student population in the northeastern USA during the spring and fall 2021 semesters, using the case-control study design.

A genetically targeted sensor reveals spatial and temporal dynamics of acrosomal calcium and sperm acrosome exocytosis.

Secretion of the acrosome, a single vesicle located rostrally in the head of a mammalian sperm, through a process known as "acrosome exocytosis" (AE), is essential for fertilization. However, the mechanisms leading to and regulating this complex process are controversial. In particular, poor understanding of Ca2+ dynamics between sperm subcellular compartments and regulation of membrane fusion mechanisms have led to competing models of AE.

Multicentric, prospective observational data show sperm capacitation predicts male fertility, and cohort comparison reveals a high prevalence of impaired capacitation in men questioning their fertility.

Research Questions: Can a previously defined relationship between sperm capacitation and the probability of a man generating pregnancy within three cycles, prospectively predict male fertility in diverse clinical settings? A second study asked, what is the prevalence of impaired sperm fertilizing ability in men questioning their fertility (MQF), and does this relate to traditional semen analysis metrics?

Design: In the multicentric, prospective observational study, data (n = 128; six clinics) were analysed to test a published relationship between the percentage of fertilization-competent, capacitated spermatozoa (Cap-Score) and probability of generating pregnancy (PGP) within three cycles of intrauterine insemination. Logistic regression of total pregnancy outcomes (n = 252) assessed fit. In the cohort comparison, Cap-Scores of MQF (n = 2155; 22 clinics) were compared with those of 76 fertile men.

Impacts of oocyte/zygote timing for in vitro fertilization and gene editing in the dog.

Previously, we reported the first live births of dogs using in vitro fertilization (IVF), embryo cryopreservation, and transfer. These techniques have potential applications in the conservation of endangered canids, and development of gene editing/repair technologies that could improve animal welfare by restoring normal gene function and removing predisposition to disease. Here, we used IVF as a springboard for initial attempts at genetic modification through gene editing/repair using the Clustered Regularly-Interspaced Short Palindromic Repeat (CRISPR)-CRISPR-associated endonuclease (Cas9) system.

Small-scale egg production centres increase children's egg consumption in rural Zambia.

Animal source foods can efficiently enhance dietary quality, but they remain inaccessible and unaffordable for many women and young children in remote, low-income communities. We piloted an intervention in which 20 groups established egg production centres (EPCs) in their rural Zambian communities to increase the availability of eggs in the local food system. In a repeated cross-sectional design over 1 year (midline [4 months after the start of egg production] and endline [11 months]), we evaluated programme impact on household egg acquisition within those communities and on egg consumption and height-for-age z score (HAZ) among young children (6-36 months) using multilevel linear, logistic, and truncated negative binomial regression techniques.

Cap-Score™ prospectively predicts probability of pregnancy.

Semen analysis (SA) poorly predicts male fertility, because it does not assess sperm fertilizing ability. The percentage of capacitated sperm determined by G localization ("Cap-Score™"), differs between cohorts of fertile and potentially infertile men, and retrospectively, between men conceiving or failing to conceive by intrauterine insemination (IUI). Here, we prospectively tested whether Cap-Score can predict male fertility with the outcome being clinical pregnancy within ≤3 IUI cycles.

Cysteine dioxygenase is essential for mouse sperm osmoadaptation and male fertility.

Sperm entering the epididymis are immotile and cannot respond to stimuli that will enable them to fertilize. The epididymis is a highly complex organ, with multiple histological zones and cell types that together change the composition and functional abilities of sperm through poorly understood mechanisms. Sperm take up taurine during epididymal transit, which may play antioxidant or osmoregulatory roles.

Timing of sperm capacitation varies reproducibly among men.

Sperm must mature functionally in the process of capacitation to become able to fertilize. Capacitation depends on membrane lipid changes, and can be quantitatively assessed by redistribution of the ganglioside G , the basis of the Cap-Score™ sperm function test. Here, differences in Cap-Score were compared among and within men at two time points.

Examining the association between livestock ownership typologies and child nutrition in the Luangwa Valley, Zambia.

Objective: To investigate the association between livestock ownership and dietary diversity, animal-source food consumption, height-for-age z-score, and stunting among children living in wildlife "buffer zones" of Zambia's Luangwa Valley using a novel livestock typology approach.

Methods: We conducted a cross-sectional study of 838 children aged 6-36 months. Households were categorized into typologies based on the types and numbers of animals owned, ranging from no livestock to large numbers of mixed livestock.

Localization patterns of the ganglioside G in human sperm are indicative of male fertility and independent of traditional semen measures.

Semen analysis lacks a functional component and best identifies extreme cases of infertility. The ganglioside G is known to have functional roles during capacitation and acrosome exocytosis. Here, we assessed whether G localization patterns (Cap-Score™) correspond with male fertility in different settings: Study 1 involved couples pursuing assisted reproduction in a tertiary care fertility clinic, while Study 2 involved men with known fertility versus those questioning their fertility at a local urology center.

Validation of a laboratory-developed test of human sperm capacitation.

Sperm must undergo capacitation to become fertilization competent. Here we validated that monosialotetrahexosylganglioside (G ) localization patterns, which were assessed in the Cap-Score™ Sperm Function Test, reflect a capacitated state in human sperm. First, we defined patterns representing sperm that do or do not respond to stimuli for capacitation.

Biomimicry Promotes the Efficiency of a 10-Step Sequential Enzymatic Reaction on Nanoparticles, Converting Glucose to Lactate.

For nanobiotechnology to achieve its potential, complex organic-inorganic systems must grow to utilize the sequential functions of multiple biological components. Critical challenges exist: immobilizing enzymes can block substrate-binding sites or prohibit conformational changes, substrate composition can interfere with activity, and multistep reactions risk diffusion of intermediates. As a result, the most complex tethered reaction reported involves only 3 enzymes.

Sustainable smallholder poultry interventions to promote food security and social, agricultural, and ecological resilience in the Luangwa Valley, Zambia.

In Zambia's Luangwa Valley, highly variable rainfall and lack of education, agricultural inputs, and market access constrain agricultural productivity, trapping smallholder farmers in chronic poverty and food insecurity. Human and animal disease (e.g.

Lipid Regulation of Acrosome Exocytosis.

Lipids are critical regulators of mammalian sperm function, first helping prevent premature acrosome exocytosis, then enabling sperm to become competent to fertilize at the right place/time through the process of capacitation, and ultimately triggering acrosome exocytosis. Yet because they do not fit neatly into the "DNA--RNA-protein" synthetic pathway, they are understudied and poorly understood. Here, we focus on three lipids or lipid classes-cholesterol, phospholipids, and the ganglioside G(M1)--in context of the modern paradigm of acrosome exocytosis.

Live Births from Domestic Dog (Canis familiaris) Embryos Produced by In Vitro Fertilization.

Development of assisted reproductive technologies (ART) in the dog has resisted progress for decades, due to their unique reproductive physiology. This lack of progress is remarkable given the critical role ART could play in conserving endangered canid species or eradicating heritable disease through gene-editing technologies-an approach that would also advance the dog as a biomedical model. Over 350 heritable disorders/traits in dogs are homologous with human conditions, almost twice the number of any other species.