[Hip fracture in the elderly: epidemiology and costs of care]. / Fractura de cadera en el adulto mayor: epidemiología y costos de la atención.

INTRODUCTION: The aim of this study was to describe the epidemiology and to estimate the direct medical costs of hip fracture among elderly patients in Mexico. MATERIAL AND METHODS: An observational, cross sectional and retrospective study was undertaken. Hospital discharge and surgical procedures for the period 2013-2018 were the databases used for the analysis, and obtained from General Directorate of Health Information. Variables included were sex, federal entity, age, year of discharge, and fracture type according to the CIE-10; and also, the supplies needed for the surgical procedures. RESULTS: A total of 16,829 patients with hip fracture were discharge, 69% were women, and the mean for age was 79 years old and for the hospital stay length was nine days. The most frequent fracture type was the femur neck with 77% and the average medical costs was USD$45,122,228.00. CONCLUSION: Falling risks increase with age, especially in patients among 80-89 years of age, hence, is expected that this type of pathology increases in the following years. The medical costs for treatment of hip fracture represents an economic impact on health services. For that reason, the implementation of prevention strategies, risk of falling for example, is the one of most efficient approach.

INTRODUCCIÓN: El objetivo general de la investigación fue describir la epidemiología y estimar los costos médicos directos de la fractura de cadera en el adulto mayor en México. MATERIAL Y MÉTODOS: Se realizó un estudio observacional y transversal retrospectivo. Se utilizaron dos bases de datos obtenidas de la Dirección General de Información en Salud del período 2013-2018: egresos hospitalarios y procedimientos quirúrgicos. Las variables incluidas fueron: sexo, entidad federativa, edad, año de registro y tipo de fractura de acorde a la CIE-10; de igual forma, todos los insumos necesarios para la realización del procedimiento quirúrgico. RESULTADOS: Se registraron 16,829 ingresos de pacientes con fractura de cadera. Las mujeres representaron 69% del total de pacientes, la edad en promedio fue de 79 años y la estancia hospitalaria fue de nueve días, 77% de las fracturas fueron de cuello de fémur y el promedio de los costos médicos directos de los procedimientos ascendieron a USD $45,122,228.00 para el período estudiado. CONCLUSIÓN: El riesgo de caídas aumenta con la edad, especialmente en el grupo etario de 80-89 años, por lo que se espera que este tipo de patologías se incremente en los próximos años. De igual forma, los costos para la atención de estas fracturas representan un impacto económico para los sistemas de salud. De manera que la implementación de estrategias de prevención, por ejemplo, en caídas es el método más eficiente para contribuir al envejecimiento saludable.

N-(2-mercaptopropionyl)-glycine enhances in vitro pig embryo production and reduces oxidative stress.

This study evaluated the effects of different concentrations (1, 10, 25, 50, and 100 µM) of the antioxidant N-(2-mercaptopropionyl)-glycine (NMPG), during the culture of in vitro-fertilized porcine oocytes. While the highest concentrations of NMPG (50 and 100 µM) were toxic to the developing embryos during the first two days of culture, 25 µM NMPG achieved cleavage rates that were similar to those achieved by the control but did not sustain blastocyst production by Day 7 of culture. Compared to the control culture medium, the culture medium supplemented with 10 µM NMPG increased (P < 0.05) the rates of blastocyst formation, decreased (P < 0.05) the intracellular levels of reactive oxygen substances, and downregulated (P < 0.05) the expression of the oxidative stress related gene GPX1. In conclusion, these results demonstrated that supplementation of porcine embryo culture medium with 10 µM NMPG can attenuate oxidative stress and increase the yield of in vitro production of blastocysts.

Effect of astaxanthin in extenders on sperm quality and functional variables of frozen-thawed boar semen.

The aim of the study was to determine whether the presence of astaxanthin (ASX) protects boar spermatozoa against damage related to cryopreservation. Pooled ejaculates extended in Beltsville Thawing Solution (BTS) were used. Three experiments were conducted: 1) sperm samples were pre-incubated overnight (17 °C) with ASX (0, 0.5, 5, 15 µM) prior to freezing and then frozen using cooling and thawing extenders supplemented with ASX (0, 0.5, 5, 15 µM); 2) sperm samples were treated with ASX (0, 0.5, 5, 15 µM) only during overnight pre-incubation (17 °C) prior to cryopreservation; and 3) a thawing extender was supplemented with ASX (0, 0.5, 5, 15 µM). The groups were as follows: control (C; no treatment), ASX 1 (0.5 µM), ASX 2 (5 µM) and ASX 3 (15 µM). Total (TM) and progressive (PM) motility was analyzed using CASA, while sperm viability, reactive oxygen species generation, lipid peroxidation and apoptoticlike changes were analyzed using flow cytometry. Sperm variables were evaluated prior to freezing as well as 30 and 150 min after thawing. In Experiment 1, the values of TM and sperm viability post-thaw were less in the ASX 3 than C group. In Experiment 2, there was no effect of ASX on any of the sperm variables evaluated, while in Experiment 3, apoptotic-like changes were less in the ASX 1 than C group. In conclusion, there was a subtle beneficial effect on cryopreserved boar spermatozoa after addition of ASX to thawing media.

Conductance-Matrix Symmetries of a Three-Terminal Hybrid Device.

We present conductance-matrix measurements of a three-terminal superconductor-semiconductor hybrid device consisting of two normal leads and one superconducting lead. Using a symmetry decomposition of the conductance, we find that antisymmetric components of pairs of local and nonlocal conductances qualitatively match at energies below the superconducting gap, and we compare this finding with symmetry relations based on a noninteracting scattering matrix approach. Further, the local charge character of Andreev bound states is extracted from the symmetry-decomposed conductance data and is found to be similar at both ends of the device and tunable with gate voltage. Finally, we measure the conductance matrix as a function of magnetic field and identify correlated splittings in low-energy features, demonstrating how conductance-matrix measurements can complement traditional single-probe measurements in the search for Majorana zero modes.

Three-to-5-day weaning-to-estrus intervals do not affect neither efficiency of collection nor in vitro developmental ability of in vivo-derived pig zygotes.

An efficient system to collect large numbers of vital zygotes is a pre-requisite for application of zygote genome-editing technology, including development of efficient models for xenotransplantation using pigs. Owing to the sub-optimal in vitro production of zygotes in pigs, efficient collection of in vivo developed zygotes is required. Timing of ovulation is a key factor to sustain efficiency since the interval between pronuclear formation and the first division is very short in pigs. The weaning-to-estrus interval can, due to its inverse relation with length of estrus and time of ovulation, interfere with ovulation and make it asynchronous, which reduces the probability of obtaining zygotes. This retrospective study compared the effects of three weaning-to-estrus intervals of 3, 4 or 5 days on zygote collection efficiency in a total of 17 trials over a 3-year period including 223 sows. Donor sows in groups of 10-15 animals were super-ovulated with eCG 24 h after weaning and those in estrus at 48-72 h post-eCG were immediately treated with hCG, followed by insemination 6 and 24 h thereafter. Collected structures during laparotomy on Day 2 (Day 0: onset of estrus) were morphologically evaluated and only those with a single cell and two visible polar bodies were considered as zygotes. Zygotes were injected with CRISPR-Cas9 editor mixture and cultured for 6 days to evaluate their developmental ability against non-injected control zygotes. Of all recovered structures (N = 5,468), 67.4%, 30.8% and 1.8% were zygotes, 2-cell embryos and oocytes-degenerated embryos, respectively. The different weaning-to-estrus intervals did not affect either the percentages of collected zygotes (range: 64.1%-70.0%) or the percentages of sows with zygotes at collection time (range: 69.0%-73.3%). The weaning-to-estrus intervals did not affect the in vitro developmental ability of zygotes. After 24 h of culture, 78.1 ±â€¯2.0% and 95.1 ±â€¯0.6 (P < 0.05) of injected (N = 2,345) and non-injected (N = 335) zygotes, respectively, developed to 2-to-4-cell embryo stage. The total efficiency of the system was 64.1 ±â€¯2.2% and 85.8 ±â€¯1.5% (P < 0.05) for injected and non-injected zygotes, respectively. In conclusion, the results indicate that neither the efficiency of collecting in vivo derived porcine zygotes from superovulated sows nor the zygote ability to develop to blastocyst after cytoplasmic genome-editing injection were affected by a weaning-to-estrus interval between 3-to-5 days.

The cytokine platelet factor 4 successfully replaces bovine serum albumin for the in vitro culture of porcine embryos.

The cytokine platelet factor 4 (PF4) enhances differentiation and cell viability of different stem cells lines in vitro. This study investigated whether PF4 addition to customary pig embryo semi-defined culture media can improve their developmental outcome (Experiment 1) and ultimately replace the need for bovine serum albumin (BSA, Experiment 2). Experiment 1 added PF4 (100-1000 ng/mL, 0 = control) to NCSU-23 with 0.4 mg/mL BSA culturing 3430 presumptive zygotes. Experiment 2 added PF4 (100-1000 ng/mL, 0 = Control-PVA) to a BSA-free medium (NCSU-23 with 0.3 mg/mL PVA) culturing 3820 presumptive zygotes. Zygote culture in NCSU-23 with 0.4 mg/mL BSA was used as overall control. All groups of Experiment 1 displayed similar rates of day 2-cleavage (range: 65.0 ±â€¯10.9 to 70.0 ±â€¯5.8%); of day 7-blastocyst rates (range: 46.6 ±â€¯10.0 to 56.4 ±â€¯8.2%) and of total day 7-blastocyst efficiency (range: 32.3 ±â€¯8.3 to 37.2 ±â€¯7.3%). Addition of PF4 did not affect total cell numbers of day 7 blastocysts (range: 44.1 ±â€¯23.2 to 50.5 ±â€¯26.4). In Experiment 2, PF4 accelerated embryo development, increasing (P < 0.01) blastocyst yield compared to 0-PF4, and blastocyst formation by day 5 adding PF4 100-500 ng/mL (range: 29.9 ±â€¯7.8 to 31.8 ±â€¯5.5%; P < 0.05) compared with BSA-control (17.2 ±â€¯8.2%) and PF4 1000 ng/mL (15.5 ±â€¯7.9%); showing similar blastocyst rates (range: 42.0 ±â€¯11.5 to 49.3 ±â€¯10.0%), total efficiency (28.0 ±â€¯8.2 to 32.3 ±â€¯7.1%) total cell numbers (range: 42.6 ±â€¯19.3 to 45.7 ±â€¯23.9) as BSA-controls. In conclusion, although PF4 did not show additive improvement under usual semi-defined, BSA-supplemented embryo media, it successfully replaced BSA sustaining porcine blastocyst production in chemically defined conditions.

Porcine blastocyst viability and developmental potential is maintained for 48 h of liquid storage at 25 °C without CO2 gassing.

Short- and medium-term storage of pig embryos has become relevant for commercial application of non-surgical deep uterine embryo transfer (NsDU-ET) in the light of the strict legal and administrative requirements posed by the International Association for Air Transport (IATA) to allow shipment of liquid nitrogen (LN2) containers and the technical drawbacks when using vitrified embryos. Therefore, this study developed an efficient method for the liquid storage of in vivo-derived porcine blastocysts for a moderate duration (48 h) without controlled CO2 gassing. We evaluated two storage temperatures (25 °C and 37 °C) and three HEPES-supplemented media: the chemically defined media TL-PVA and NCSU-PVA and the semi-defined medium NCSU-BSA. We observed no differences in survival, hatching rate or final developmental stage between the two temperatures, but storage at 25 °C was more efficient to preserve zona pellucida (ZP) integrity. Blastocysts were successfully stored for 24 h in a chemically defined medium. Yet, only 48 h storage in NCSU-BSA medium supported blastocyst development. Although all storage conditions resulted in an embryonic developmental delay, blastocysts stored in NCSU-BSA at either tested temperature could hatch and attain the same final developmental stage as control blastocysts when cultured under standard conditions after storage. Moreover, blastocysts stored at 25 °C for 48 h in NCSU-BSA medium could produce pregnancies after surgical transfer. In conclusion, porcine blastocysts maintain their viability and developmental potential after storage in the semi-defined medium NCSU-BSA for at least 48 h at 25 °C.

Boar semen proteomics and sperm preservation.

Recently numerous proteomic approaches have been undertaken to identify sperm and seminal plasma (SP) proteins that can be used as potential biomarkers for sperm function, including fertilization ability. This review aims firstly to briefly introduce the proteomic technologies and workflows that can be successfully applied for sperm and SP proteomic analysis. Secondly, we summarize the current knowledge about boar SP and the sperm proteome, focusing mainly on its relevance to sperm preservation procedures (liquid storage or cryopreservation) and their outcomes in terms of sperm function and fertility.

High pre-freezing sperm dilution improves monospermy without affecting the penetration rate in porcine IVF.

The high incidence of polyspermy is still an unresolved problem for the production of in vitro-produced porcine embryos. In this work, we modified the usual sperm processing sequence for in vitro fertilization (IVF), and the spermatozoa from four boars were frozen directly at a low sperm concentration of 20 × 106 sperm/mL (high pre-freezing sperm dilution group; F20), thawed and processed for IVF in three replicates. Spermatozoa from the same boars frozen at a conventional concentration (1000 × 106 sperm/mL) were used as the control group. The post-thaw sperm quality evaluation demonstrated that despite there being no differences in the percentage of motile spermatozoa between groups, the proportion of live spermatozoa with intact acrosomes was significantly higher in the F20 group than in the control. The in vitro penetration rate was also similar between groups; however, the co-incubation of oocytes with F20 sperm increased monospermy, IVF efficiency, cleavage rate and the efficiency of blastocyst formation compared with the results for oocytes co-incubated with control spermatozoa. These results indicate, for the first time, that a high pre-freezing sperm dilution increases monospermy without affecting penetration rates, thereby increasing blastocyst formation.

Exogenous ascorbic acid enhances vitrification survival of porcine in vitro-developed blastocysts but fails to improve the in vitro embryo production outcomes.

In this study, the effects of addition of the antioxidant ascorbic acid (AsA) were evaluated during porcine in vitro embryo production (IVP) and vitrification. In experiment 1, the effects of AsA supplementation during IVM, IVF and IVC were evaluated, using a total of 2744 oocytes in six replicates. The IVM, IVF and embryo IVC media were supplemented or not (control) with 50 µg/mL AsA in all possible combinations. No significant effects of AsA were detected in any of the maturation, fertilization or embryo development parameters assessed. In experiment 2, we evaluated the effects of adding AsA to vitrification-warming media on the post-warming survival and quality of blastocysts. Day-6 in vitro-produced blastocysts (N = 588) from six replicates were randomly divided in two groups, with vitrification and warming media either supplemented with 50 µg/mL AsA (VW + group) or un-supplemented (VW- control). Addition of AsA increased (P < 0.05) blastocyst survival rate after vitrification compared with that of VW- control embryos. Vitrification and warming increased (P < 0.05) the production of oxygen species (ROS) and reduced (P < 0.05) the glutathione levels in blastocysts. Although VW + blastocysts displayed higher (P < 0.05) ROS levels than those of fresh control blastocysts, the levels were lower (P < 0.05) than those found in VW- control blastocysts. In conclusion, under the experimental conditions, supplementation of IVM/IVF/IVC media with AsA did not improve the embryo production in vitro. By contrast, the addition of AsA to chemically defined vitrification and warming media increased the survival of in vitro-produced porcine blastocysts by decreasing ROS production.

The effects of multiple anaesthetic episodes on equine recovery quality.

BACKGROUND: Although rare, 70% of equine fatalities during recovery from general anaesthesia (GA) are due to catastrophic fractures from poor recovery quality. OBJECTIVE: To determine the effect of repeated GA recovery on GA recovery quality. STUDY DESIGN: Experimental blinded trial. METHODS: Eight adult horses underwent six GA events on sevoflurane for distal limb MRI examination over a 14-week period. Prior to GA recovery, xylazine was administered. Randomly ordered video-recorded GA recoveries were scored by three blinded board certified veterinary anaesthesiologists, unaware of patient identity or GA event number, for nine parameters using a 100 mm visual analogue scale (VAS) where 0 = worst and 100 = best. The number of attempts to stand, duration of lateral and sternal recumbency, total recovery duration and physiologic parameters during each GA event were recorded. Repeated measures ANOVA were used to detect differences. Agreement between observer VAS scores was determined via inter-rater reliability using an intraclass correlation. RESULTS: With GA recovery experience, VAS scores for balance and coordination, knuckling, and overall quality of recovery were improved and the duration of lateral recumbency was increased. There were no differences in total recovery duration, number of attempts to stand, physiologic parameters other than heart rate during GA, or VAS scores for activity in lateral recumbency, move to sternal, move to stand, or strength. MAIN LIMITATIONS: Each GA event was relatively short and there was no surgical stimulation. The same results may not occur if there was surgical stimulation and pain during each GA event. CONCLUSION: Recovery from GA improves with multiple anaesthetic episodes in horses. Clinicians can advise clients that horses are likely to have better GA recovery on repeated GA recovery due to improved balance and coordination and reduced knuckling. Additionally, there is no change in anaesthetic morbidity with six repeated GA events over a 14-week period.

Importance of oil overlay for production of porcine embryos in vitro.

Technologies to edit the zygote genome have revolutionized biomedical research not only for the creation of animal models for the study of human disease but also for the generation of functional human cells and tissues through interspecies blastocyst complementation technology. The pig is the ideal species for these purposes due to its great similarity in anatomy and physiology to humans. Emerging biotechnologies require the use of oocytes and/or embryos of good quality, which might be obtained using in vitro production (IVP) techniques. However, the current porcine embryo IVP systems are still suboptimal and result in low monospermic fertilization and blastocyst formation rates and poor embryo quality. During recent years, intensive investigations have been performed to evaluate the influence of specific compounds on gametes and embryos and to avoid the use of undefined supplements (serum and serum derivate) in the incubation media. However, little consideration has been given to the use of the mineral oil (MO) to overlay incubation droplets, which, albeit being a routine component of the IVP systems, is a totally undefined and thus problematic product for the safety of gametes and embryos. In this review, we provide an overview on the advantages and disadvantages of using MO to cover the incubation media. We also review one important concern in IVP laboratories: the use of oils containing undetected contamination. Finally, we discuss the effects of different types of oils on the in vitro embryo production outcomes and the transfer of compounds from oil into the culture media.

Simple storage (CO2-free) of porcine morulae for up to three days maintains the in vitro viability and developmental competence.

The advancement of porcine embryo transfer (ET) technology is constrained by regulatory hurdles (liquid nitrogen transportation) or, more importantly, the technical obstacles of using vitrified embryos in combination with nonsurgical deep uterine ET technology. Maintaining embryos in culture during transport and prior ET collides with the need of CO2 gassing and the best choice of culture medium. In this work, we describe storage conditions for short-term embryo CO2-free storage that allowed for a majority of in vivo-derived porcine morulae to survive after 3 days of storage in a liquid state, and to develop to the blastocyst stage unhatched, a sanitary prerequisite for ET. The storage conditions included NCSU-23 medium supplemented with bovine serum albumin, where bicarbonate was partially replaced by HEPES to avoid the need for CO2 gassing, and a temperature of 37 °C. These conditions were able to maintain the functionality of the stored embryos (hatching capacity after exposure to conventional culture conditions) and their developmental competence after ET (normal fetuses by day 38 of pregnancy). Use of this strategy of CO2-free storage should allow the shipment of fresh embryos worldwide in the absence of liquid nitrogen.

Altrenogest treatment before weaning improves litter size in sows.

The study aimed to assess whether altrenogest treatment, fed before weaning (from -8 to -2 days), could improve fertility of sows showing reproductive seasonality. Ninety sows (50 in winter-spring [WS] and 40 in summer-autumn [SA]) were randomly selected and assigned to control (C; 27 in WS and 20 in SA) or altrenogest treatment (A; 23 in WS and 20 in SA) groups. The diameter and number of ovarian follicles were transrectally scanned at the onset of oestrus. Oestrus was evaluated twice daily from weaning to day 8 post-weaning. Sows in oestrus were post-cervically inseminated at 0 and 24 hr after the onset of oestrus with liquid stored semen (1.5 × 109 sperm/doses), and farrowing rates (FR) and total piglets born (LS) were recorded. More (p < .01) sows showed no signs of oestrus within 8 days after weaning in SA (30%) than in WS (2%), without differences between A and C groups. The diameter (cm) of the follicles at the onset of oestrus was larger in A than in C sows (0.76 ± 0.01 vs 0.73 ± 0.01; p < .01), irrespective of the season. No differences in the number of follicles were found. FR did not differ between seasons and groups, being always above 85%. LS was larger (p < .01) in A (14.00 ± 0.46) than C (12.27 ± 0.44) sows, irrespective of the season. In conclusion, a short-term altrenogest treatment at the end of lactation improves the total number of piglets born from weaned sows, probably by promoting a better and more homogeneous follicular development at the start of oestrus.

Peroxidized mineral oil increases the oxidant status of culture media and inhibits in vitro porcine embryo development.

The use of oils with undetected alterations is a long-recognized problem for in vitro embryo production systems. Since peroxides in oils have been associated with reduced embryo production outcomes, our goals were (1) to evaluate the effects of a batch of mineral oil (MO) that was suspected to be altered on the in vitro production of pig embryos and (2) to determine oil peroxide values throughout culture and the transfer of oxidant agents from oil to culture media. Sunflower oil, which has a completely different chemical composition than MO but a higher oxidative status, and unaltered MO were used as controls. Oocyte maturation, fertilization and embryo development were affected differently depending on the oil overlay used. While the suspected MO was not able to sustain in vitro maturation and fertilization, the oocytes incubated in the presence of sunflower oil were matured and fertilized similarly to those of the unaltered MO group. Moreover, the cleavage rate of presumed zygotes cultured under the suspected MO was severely reduced compared with those cultured under the other oils, and none of the cleaved embryos developed to the blastocyst stage. Although the cleavage rates in the sunflower oil and unaltered MO groups were similar, embryos cultured under sunflower oil also failed to develop to the blastocyst stage. Our results revealed that the suspected MO and sunflower oil had similar levels of peroxides and that these levels were much higher than those of the unaltered MO. The total oxidant status was higher in media incubated under peroxidized oils than in fresh media or media incubated without an oil overlay or under unaltered MO, indicating that oxidant agents were transferred to the incubation media. However, unlike the sunflower oil group, the culture media incubated under the suspected MO had high levels of total oxidant status and low levels of hydrogen peroxide and reactive oxygen species, suggesting the presence of other unknown oxidant agents in that oil. These results indicate that a peroxidized MO overlay dramatically decreases embryo production outcomes. This decrease could be associated with the higher peroxide values of the oil but cannot be explained by the levels of hydrogen peroxide and reactive oxygen species transferred from the oil to the culture media. It is likely that different oxidant agent(s) and/or other toxic compounds present in the peroxidized MO are responsible for its damaging effects on oocytes and embryos.

Effects of meiotic inhibitors and gonadotrophins on porcine oocytes in vitro maturation, fertilization and development.

This study evaluated the effect of three reversible meiotic inhibitors (MINs) and their interaction with gonadotrophins (Gns) on the meiotic maturation and developmental competence of porcine oocytes. In experiment 1, the oocytes were matured for 22 hr in the presence or absence of dbcAMP (1 mM), cycloheximide (7 µM) or cilostamide (20 µM) with or without Gns, and for an additional 22 hr in the absence of MINs and Gns. At 22 hr of maturation, regardless of the presence of Gns, a higher proportion (p < .001) of oocytes cultured in the presence of MINs were effectively arrested at the germinal vesicle stage compared with the oocytes cultured without MINs. At 44 hr of maturation, the proportion of oocytes that reached MII was higher (p < .05) in groups with Gns compared with groups without Gns. In experiment 2, oocytes that were matured as in experiment 1 were inseminated and cultured for 7 days to evaluate fertilization parameters and blastocyst formation. Only oocytes from the dbcAMP + Gns group had higher (p < .05) efficiency of fertilization compared with the other treatment groups. The presence of dbcAMP during maturation also increased (p < .05) blastocyst formation and efficiency of blastocyst formation in both the presence and absence of Gns. These results indicate that the interaction of Gns with the tested MINs improved meiotic progression. In addition, regardless of supplementation with Gns, the presence of dbcAMP during the first maturation period increased and even doubled the capacity of oocytes to develop to the blastocyst stage.

Profile and reproductive roles of seminal plasma melatonin of boar ejaculates used in artificial insemination programs.

Melatonin (MLT) is present in seminal plasma (SP) of mammalian species, including pigs, and it is credited with antioxidant properties. This study aims to identify the sources of variation and the role of boar SP MLT on sperm quality and functionality and in vivo fertilizing ability of liquid-stored semen doses used in AI programs. The SP MLT was measured using an ELISA kit in a total of 219 ejaculates collected from 76 boars, and reproductive records of 5,318 AI sows were recorded. Sperm quality was assessed according to motility (computer-aided sperm analysis) and viability (cytometry evaluation). Sperm functionality was assessed according to the cytometric determination of intracellular HO generation, total and mitochondrial O production, and lipid peroxidation in liquid AI semen samples stored at 17°C over 144 h. The concentration of SP MLT differed among seasons ( < 0.01) and day length periods ( < 0.001) of the year, demonstrating that the ejaculates collected during the increasing day length period (9.80 ± 1.38 pg/mL, range: 2.75-21.94) had lower SP MLT concentrations than those collected during the decreasing day length period (16.32 ± 1.67 pg/mL, range: 5.02-35.61). The SP MLT also differed ( < 0.001) among boars, among ejaculates within boar, and among portions within the ejaculate, demonstrating that SP from the first 10 mL of sperm-rich ejaculate fraction (SRF) exhibited lower MLT concentrations than post-SRF. The SP MLT was negatively related ( < 0.001) to mitochondrial O production in viable sperm. The SP MLT did not differ among AI boars ( = 14) hierarchically grouped according to high and low fertility outcomes. In conclusion, SP MLT concentration in AI boars varies depending on the season of ejaculate collection and differs among boars, ejaculates within boar, and portions within ejaculate. The SP MLT may act at the mitochondrial level of sperm by reducing the generation of O. However, this antioxidant role of SP MLT was not reflected in sperm quality or in vivo fertility outcomes of AI semen doses.

Factors of importance when selecting sows as embryo donors.

The improvement in porcine embryo preservation and non-surgical embryo transfer (ET) procedures achieved in recent years represents essential progress for the practical use of ET in the pig industry. This study aimed to evaluate the effects of parity, weaning-to-estrus interval (WEI) and season on reproductive and embryonic parameters at day 6 after insemination of donor sows superovulated after weaning. The selection of donor sows was based on their reproductive history, body condition and parity. The effects of parity at weaning (2 to 3, 4 to 5 or 6 to 7 litters), season (fall, winter and spring), and WEI (estrus within 3 to 4 days), and their interactions on the number of corpus luteum, cysts in sows with cysts, number and quality of viable and transferable embryos, embryo developmental stage and recovery and fertilization rates were evaluated using linear mixed effects models. The analyses showed a lack of significant effects of parity, season, WEI or their interactions on any of the reproductive and embryonic parameters examined. In conclusion, these results demonstrate that fertilization rates and numbers of viable and transferable embryos collected at day 6 of the cycle from superovulated donor sows are not affected by their parity, regardless of the time of the year (from fall to spring) and WEI (3 or 4 days).

Surgical embryo collection but not nonsurgical embryo transfer compromises postintervention prolificacy in sows.

Recent advances in nonsurgical deep uterine (NsDU) embryo transfer (ET) technology allow the noninvasive transfer of porcine embryos into recipients, overcoming the most important impediment for commercial ET in this species. Although many factors in the porcine ET-field have been recently evaluated, many others remain to be explored. We investigated here the future reproductive performance of donors and recipients after artificial insemination subsequent to the default surgical embryo recovery approach and to the NsDU-ET procedure, respectively. Although surgical embryo collection did not influence subsequent farrowing rates (90.5%), litter size decreased severely (8.9 ± 0.8 piglets) compared to presurgery (10.8 ± 0.3 piglets) and control group (10.7 ± 0.3 piglets). In contrast, NsDU-ETs did neither affect fertility nor prolificacy of recipients in the cycle subsequent to ET, regardless of whether they were pregnant after NsDU-ET or not. These results indicate that while the surgical embryo collection procedure compromises the reproductive future of donor sows, the NsDU-ET approach does not affect the reproductive potential of the recipients after reintroduction to the breeding stock of the farm. Further research is thus needed to improve surgical embryo collection.

Generation of human organs in pigs via interspecies blastocyst complementation.

More than eighteen years have passed since the first derivation of human embryonic stem cells (ESCs), but their clinical use is still met with several challenges, such as ethical concerns regarding the need of human embryos, tissue rejection after transplantation and tumour formation. The generation of human induced pluripotent stem cells (iPSCs) enables the access to patient-derived pluripotent stem cells (PSCs) and opens the door for personalized medicine as tissues/organs can potentially be generated from the same genetic background as the patient recipients, thus avoiding immune rejections or complication of immunosuppression strategies. In this regard, successful replacement, or augmentation, of the function of damaged tissue by patient-derived differentiated stem cells provides a promising cell replacement therapy for many devastating human diseases. Although human iPSCs can proliferate unlimitedly in culture and harbour the potential to generate all cell types in the adult body, currently, the functionality of differentiated cells is limited. An alternative strategy to realize the full potential of human iPSC for regenerative medicine is the in vivo tissue generation in large animal species via interspecies blastocyst complementation. As this technology is still in its infancy and there remains more questions than answers, thus in this review, we mainly focus the discussion on the conceptual framework, the emerging technologies and recent advances involved with interspecies blastocyst complementation, and will refer the readers to other more in-depth reviews on dynamic pluripotent stem cell states, genome editing and interspecies chimeras. Likewise, other emerging alternatives to combat the growing shortage of human organs, such as xenotransplantation or tissue engineering, topics that has been extensively reviewed, will not be covered here.