Reproductive Purposes Research Articles

Legal regime of unimplanted embryos in vitro from the positions of doctrine, civil law and judicial practice

The article deals with the problem of determining the legal regime of unimplanted embryo in vitro, which due to the significantdevelopment of the field of assisted reproductive technologies, is increasingly at risk of its illegal use, or even illegal creation for commercialor other non-infertility treatment. The author analyzes different doctrinal approaches in the civil law of Ukraine and Germany,through the prism of the current legal regulation and practice of the European Court of Human Rights. The paper supports the positionon the right to the embryo as a personal immaterial right and expands it in the sense that such a component of reproductive rights asthe right to determine the future fate of embryos in vitro will also belong to persons to whom assisted reproductive technologies havebeen applied and as a result these embryos where created. Resulting from this analysis, the position on the inadmissibility of the interpretationof the unimplanted embryo as an object of property law is expressed. To support this point of view, the author cites the judgmentof the European Court of Human Rights in Parrillo v. Italy, in which the court emphasized the inadmissibility of the assessmentof embryos as an object of property rights. In particular, the paper proposes to define the regime of unimplanted embryo in vitro as anobject that has a personal immaterial connection to persons for whose treatment of infertility (parents-customers) it was created. Andit is these individuals who will have the right to determine its future. In this regard, it is proposed to reflect this concept in the cuurentregulation in partricular Procedure for the use of assisted reproductive technologies. From the proposed wording, first, it will followthat in vitro embryos created as a result of the partial or full use of donor biological material at the request of persons to whom assistedreproductive technologies are applied will have this personal connection only with the future parents and there will not be an ethicaldilemma regarding who will have the right to determine their future fate (biological parents (reproductive cell donors) or future parents).Secondly, it will also mean that embryos can only be created for reproductive purposes, and an institution providing reproductive ser -vices will not have the right to create or dispose of embryos in vitro at its own discretion without the proper consent of its future parent.And, thirdly, it will exclude the possibility of interpreting such embryos as objects of property rights.

The use of human artificial gametes and the limits of reproductive freedom.

Recent developments in generating gametes via in vitro gametogenesis (IVG) from induced pluripotent stem cells (iPSCs) and their successful use for reproductive purposes in animals strongly suggest that soon these methods could also be used in human reproduction. At least two questions emerge in this context: (a) if a legislator should permit their use and (b) if ethical claims emerge that support their provision, e.g., by public health care systems. This urges an ethical reflection of the new reproductive options this technique might offer. Since the concept of reproductive freedom is a key aspect for the ethical evaluation of artificial reproductive technologies (ARTs), it is necessary to analyze if the new possibilities emerging from IVG fall within the scope of this concept. The results may constitute a morally relevant difference between different imaginable applications of IVG and potentially justify differences in claims to access this technology.

DNA methylation status of bovine blastocysts obtained from peripubertal oocyte donors.

In the dairy industry, the high selection pressure combined with the increased efficiency of assisted reproduction technologies (ART) are leading toward the use of younger females for reproduction purposes, with the aim to reduce the interval between generations. This situation could impair embryo quality, decreasing the success rate of the ART procedures and the values of resulting offspring. Young Holstein heifers (n = 10) were subjected to ovarian stimulation and oocyte collection at 8, 11, and 14 months of age. All the oocytes were fertilized in vitro with semen from one adult bull, generating three pools of embryos per animal. Each animal was its own control for the evaluation of the effects of age. The EmbryoGENE platform was used to compare the DNA methylation status of blastocysts obtained from oocytes collected at 8 versus 14 and 11 versus 14 months of age. Age-related contrast analysis identified 5,787 and 3,658 differentially methylated regions (DMRs) in blastocysts from heifers at 8 versus 14 and 11 versus 14 months of age, respectively. For both contrasts, the DMRs were distributed nonrandomly in the different DNA regions. The DNA from embryos from 8-month-old donors was more hypermethylated, while the DNA from embryos from 11-month-old donors displayed an intermediate phenotype. According to Ingenuity Pathway Analysis, the upstream regulator genes cellular tumor antigen p53, transforming growth factor β1, tumor necrosis factor, and hepatocyte nuclear factor 4α are particularly associated with methylation sensitive targets, which were more hypermethylated in embryos from younger donors.

Nest construction and presence do not alter territorial aggression in male threespine stickleback

Constructing a nest within a mating territory provides a clear benefit to the resident, particularly by improving the opportunity to mate. It is unclear whether animals who use nests exclusively for reproductive purposes account for either the effort invested or the resulting increase in mating potential when valuing their territories. We sought to explicitly reveal a nest’s added subjective resource value through within-group comparison of aggressive behaviour before and after nest construction. An increase in aggression following construction would indicate that the resident perceives greater subjective value in his territory, and thus values the nest. Threespine stickleback, Gasterosteus aculeatus, fish demonstrate stereotypical aggressive behaviours during an easily induced territorial defence. The male’s nest is used exclusively for reproductive purposes, avoiding any confound of shelter. Contrary to our hypothesis, neither nest presence, timing of construction, nor nesting outcome was associated with differences in behavioural measures of territorial aggression. Assessed behaviours were robust, repeatable and inter-correlated. We conclude that territorial aggression is neither predictive of nor altered by nesting in threespine stickleback fish. Our results suggest that nests used transiently for a portion of the mating season add negligible subjective resource value to a territory. We additionally demonstrate that examinations of territorial aggression in sticklebacks do not need to control for nest building, improving statistical power by decreasing dropout rates. These results dovetail with recent work in other fish species to suggest that assessment of territorial aggression absent a nest may be practicable for fish in general.

The Beginning of Life Issues: An Islamic Perspective.

Islam gives legal precedence to purity of lineage and known parenthood of all children. In Islam treatment to infertility using IVF is permitted within validity of marriage contract with no genes mixing. The paper shows that the Qur'ān, the word of Allah, and science, the deeds of Allah are not in major conflicts in defining the start of human life. The Holy Qur'ān provides an elegant description of origin, developmental stages of intra-uterine life. The Hadith explains two positions one that believes human embryo get ensouled at conception and the other after 40days of conception. The paper aims to find that Islam confers moral respect to human embryo, but it also clarifies the absence of full human rights to a developing foetus. In Islam, human embryonic use is probably permissible for therapeutic and reproductive purpose keeping intact the principles of Shari'ah.

Impact of a national genetic carrier-screening program for reproductive purposes.

The Israeli population, encompassing 9 million citizens, is comprised of diverse communities. The Ministry of Health genetic screening program for reproductive purposes was introduced in 2013. This program is mainly aimed at severe incurable diseases with high rates of infant and childhood morbidity and/or mortality, with a carrier frequency of at least 1:60 and/or a disease frequency of 1 in 15000 live births. In this paper, we present the results of the national genetic carrier-screening program implementation. Data acquisition for this study was performed by retrospectively searching Ministry of Health database, which includes the reports of 18 genetic laboratories performing genetic screening tests. During 2015-2017, a total of 919820 carrier-screening genetic tests were executed. The overall number rose by 14.9% over these years. For about two-thirds of the presented disorders, carrier frequency was within the expected range. A decrease of 57% was noted in the observed number of patients with spinal muscular atrophy born during 2014-2017, compared with the expected rate. Familial dysautonomia, Canavan and Tay-Sachs diseases yielded a very low prevalence. Our results highlight the impact of a national genetic carrier-screening program. Couples at risk of an affected fetus mostly choose to perform preconception or prenatal diagnosis and to act accordingly. Our country has several characteristics enabling us to achieve this success, including considerable rates of endogamy and consanguineous marriages, increased frequency of founder mutations, and high fertility rates. In addition, wide accessibility of the tests and good compliance of the population must be noted. Still, raising the awareness and continuing education of population and caregivers about the importance and efficiency of carrier screening remains an important issue. Finally, expanding the existing tests into a uniform, wide genetic panel seems to be the next goal.

Posthumous planning following fertility preservation: a study of adolescent cancer patients in Israel

In an Israeli qualitative study with adolescent cancer survivors and parents who had considered fertility preservation, practically all participants could not recall any discussions with healthcare providers about plans for cryopreserved biological materials in the case of death. This finding is surprising given recent court struggles in Israel over the posthumous use of cryopreserved sperm. In interviews with these adolescent survivors and their parents, intended future use of cryopreserved biological materials is directed for affected individuals’ reproductive purposes later in life, with hardly any consideration of others' use of these materials for posthumous reproduction. To avoid future ethical and legal quagmires, healthcare professionals should have frank discussions with adolescents and their parents about what to do with such materials in the case of death. This paper discusses the socio-cultural and legal Israeli context and considers the ethical implications of using minors’ biological materials for posthumous assisted reproduction in Israel.

The in Vitro Embryo and the Law: The Ownership Issue and a Response to Robinson

In 2012 the Minister of Health made the Regulations Relating to the Artificial Fertilisation of Persons, which provide that the woman who intends to be made pregnant with an in vitro embryo owns such an embryo and can control the embryo's fate in specified ways. Given that in vitro embryos are outside the woman's body, the rationale for these provisions cannot be to protect the woman's bodily integrity. These provisions are, however, problematic from a constitutional perspective, as they: exclude fathers across the board, and impede the right of all intended parents who will not gestate the pregnancy, like surrogacy commissioning parents, to make decisions regarding reproduction – which include the right not to reproduce and hence to veto the further use of an in vitro embryo for reproductive purposes. Robinson argues that the legislative intent with the 2012 Regulations was not to establish ownership of in vitro embryos, and that in vitro embryos are not legal objects (or subjects), but rather form part of the legal subjectivity of their parents. I respond that the language used in the relevant provision is plain and clear in establishing ownership of in vitro embryos, and that in vitro embryos are therefore legal objects. I further suggest that Robinson's proposition of in vitro embryos forming part of the legal subjectivity of their parents may address the gender equality concern with the 2012 Regulations, but that it in turn causes other problems. In particular, Robinson's rationale for his proposition is problematic, as it appears to conflate the embryo with the prospective child. I rely on the important recent judgment in Ex Parte KAF 2019 2 SA 510 (GJ) that held explicitly that the in vitro embryo should not be equated with the prospective child. Finally, I respond to Robinson's critique of my 2005 article, by clarifying the research questions and answers of that article. I highlight the importance of the moral status of the in vitro embryo to legal and ethical debates relating to the in vitro embryo, and invite academic debate on the topic.

Reproductive biology of Ophelia barquii (Annelida, Opheliidae) along the Salento Peninsula (Mediterranean Sea, South Italy)

This study investigates two populations of a polychaete identified as Ophelia barquii Fauvel, 1927, collected in two beaches from the Adriatic (Alimini beach) and Ionian (Gallipoli beach) coasts from the Salento Peninsula. Each population was analyzed monthly from April 2017 to March 2018. In both beaches, populational density decreased from April to August with a complete absence of worms from August to the following January. We hypothesize that this trend results from horizontal migration of individuals for reproductive purposes. Sexual maturation was asynchronous between the two populations, with individuals becoming ripe earlier in Alimini than in Gallipoli. Significant differences in body size were recorded between the two populations, with specimens larger in Gallipoli than in Alimini, suggesting that they could belong to different species. However, life-cycle studies performed under laboratory conditions showed that both populations follow a similar developmental path and can cross-fertilize. The larval development featured a very short period of pelagic life, the shortest known so far for any species of Ophelia.

Upper Cretaceous Benthic Foraminifera Thriving in Intertidal Areas of the Apennine Carbonate Platform

The intertidal area of carbonate platform hosts a complex array of ecological networks in which microbials, algae, cyanobacteria and benthic foraminifera coexist. Being influenced by the amplitude of tides, intertidal areas frequently experience episodes of extreme conditions, including hypersaline waters and elevate daily temperatures. An interesting record of Upper Cretaceous fossil benthic foraminifera found within the intertidal facies of Apennine Carbonate Platform is presented and discussed. Two local gatherings of juvenile Scandonea and adult Rotalispira maxima suggest that the intertidal area was frequently colonized by benthic foraminifera. The results show that these populations of foraminifera were able to tolerate periods of extreme salinity and temperature for reproduction or feeding purposes.

Ultrasonography of the liver in healthy and diseased camels (Camelus dromedaries).

In camels, hepatic diseases are relatively common and most of them are misdiagnosed as a cause of illness because signs may be subtle. In addition, diagnostic laboratory methods are insufficient as hepatic enzymes can also be elevated in camels with cardiac or skeletal muscle damage. Examples of liver diseases in camels are hepatic lipidosis, hepatitis, cirrhosis, hepatic necrosis, choleostasis, hyperplasia of biliary epithelium, hydatid cysts, glycogen deposition, cholangitis, cholangiohepatitis, calcified hydatid cyst and hepatic abscesses. When the liver is examined by ultrasonography, the clinician gets sufficient information about the size, position, echopatterns of the hepatic parenchyma, bile ducts and outlines of the hepatic blood vessels. Ultrasonography has been used previously in camels only for reproductive purposes. However, during the past decade, it has been used for scanning of the healthy organs as well as evaluation and determining the diagnosis and prognosis of non-reproductive disorders. Examples of diseases evaluated by ultrasonography in camels are paratuberculosis, trypanosomiasis, abdominal and urinary disorders, thoracic diseases, renal tumors, pyelonephritis, renal abscessation, gastrointestinal tumors, chronic peritonitis and splenic abscessation. Ultrasound-guidance in biopsy of hepatic lesions and in portocentesis has also been reported in camels. This mini review article is written to shed light on ultrasonography of the liver and its blood vessels in healthy camels as well as finding in camels with hepatic disorders such as fatty infiltration of the liver, hepatic abscesses and calcification of the bile ducts.

Focusing on Human Rights: a framework for CRISPR germline genome editing ethics and regulation.

The late 2018 announcement of the claimed births of CRISPR-edited babies has stimulated widespread condemnation and calls by some leading scientists for a moratorium on any further germline genome editing (GGE) for reproductive purposes. Concurrently, national and international bodies are calling for the development of robust guidelines and regulations that will identify permissible conditions under which such GGE efforts might eventually proceed. Crucially, these conditions go beyond rigorous safety standards to address some of the social and ethical concerns that arise with germline interventions. As these bodies convene to navigate this unique terrain, we suggest an important standard for generating ethically robust guidelines. Our approach builds from concerns about social exclusion and social justice with a focus on fundamental human rights. We believe that a deontological or rights-based approach, rather than a utilitarian approach, is needed to ensure that this socially disruptive technology minimizes further marginalization of people with disabilities and does not create a new form of social injustice. In pursuit of a deontological framework, we propose the implementation of an objective assessment tool: the Human Rights Impact Assessment (HRIA). Use of the HRIA establishes necessary constraints on applications of GGE in order to safeguard the most vulnerable members of society.

Hysteropexy in the Treatment of Total Uterine Prolapse in a Brazilian Molosser Bitch

Background: Uterine prolapse is an obstetric emergency because the exposed tissue can present edema, devitalization or even gangrene, depending on the evolution time and on the severity of circulatory changes. Haemorrhage due to the rupture of ovarian or uterine vessels can evolve rapidly to hypovolemic shock. More rarely, secondary thromboembolism to the uterine strangulation can be developed, complication that can also lead to death. This study reports the case of a Brazilian Molosser bitch treated by reducing total uterine prolapse, complemented with hysteropexy, in order to prevent recurrence and maintain its reproductive ability. Case: A 5-year-old Brazilian Molosser bitch was attended presenting double firm and tubular mass protruding through the vulva. The patient gave birth to five live pups, the last two of which needed to be pulled. Two days after parturition, she had abdominal contraction when a mass protruded from the vagina. A total uterine prolapse was diagnosed, since both uterine horns were found exposed and inverted, with exposition of the endometrium and the areas of placental implantation, together with the body of the uterus. The tissue was apparently edematous and lightly parched, with the presence of strange bodies, but without signals of circulatory complications or lacerations that could turn a possible reduction and maintenance of the uterus unfeasible. Due to interest of the tutor in preserving the reproductive ability, an internal reduction via ventral medial celiotomy and the fixation of the uterus in the lateral abdominal wall (hysteropexy) was chosen in order to diminish the possibility of relapse in future parturition.Discussion: The definitive treatment of the uterine prolapse can be realized through ovariohysterectomy when the exposed uterine tissue is highly compromised or there is vessel rupture or no reproductive purpose for the female. The amputation of everted tissue through the vulva is also an option, but the risk of intracavitary bleeding during tissue resection must be considered when the uterine and ovarian artery and vein escape to the interior of the abdomen without according hemostasis. In cases which there are no complications of the prolapsed tissue and is desired to maintain the reproductive ability of the female, the internal reduction with hysteropexy is recommended. As the etiology of the uterine prolapse is a condition bound to several factors, in between them multiple pregnancies, relaxation of the uterine ligaments and genetic factors, conditions which can be associated to the cause of the prolapse in the mentioned patient, the hysteropexy was preferred as a complementary technique to uterine preservation and prevention of the prolapse relapse in a possible new parturition. In this case, the reduction of prolapse followed by hysteropexy was preferred, because the female dog was apparently well, without signs of complication of the prolapsed tissue and with satisfactory preoperative evaluation. Adding these to the request of the tutor in maintaining the reproductive ability of the female, since the procedure would not pose as a life risk to it. The absence of severe lacerations and circulatory complications in the prolapsed uterus of the bitch under study and the non-detection of toxemia and septicemia corroborated to the reduction and preservation of the uterus. Even though the uterine prolapse is a rare disease, it is necessary to improve surgical techniques that enable the conservation of the female dog in reproduction. In situations of viability of the prolapsed tissue, the technique of hysteropexy in abdominal wall showed itself effective, concluding that hysteropexy is feasible and efficient as a complementary procedure to the reduction of uterine prolapse, having contributed to avoid relapse and maintaining the reproductive of the subject related in this article.

Donantes de ovocitos: análisis comparativo de dos muestras de Argentina y España sobre perfil de donantes, motivaciones y anonimato

La ovodonación es el proceso por el cual una mujer dona óvulos a una clínica de reproducción asistida o a un banco de gametos con fines reproductivos (para una mujer receptora) o con la finalidad de utilizarlos para investigación biomédica. Los ciclos de tratamiento donde intervienen óvulos “donados” son cada vez más frecuentes en todo el mundo. España ocupa el primer lugar en Europa en donación de ovocitos de acuerdo a los datos de 2016 de la European Society of Human Reproduction and Embryology (ESHRE). En 2016 se realizaron en Argentina 13.823 ciclos de tratamiento, con 2709 transferencias de ovodonación (19,5%) (SAMeR, 2017). El presente estudio tiene como objetivo visibilizar los perfiles sociodemográficos de dos muestras de donantes de ovocitos de Argentina (n=93) y de España (n=49), las diferencias si es que existen, en las motivaciones que presentan las donantes para inscribirse en los programas de donación de ovocitos y sus actitudes hacia el anonimato. La metodología del estudio es cuantitativa, mientras que las inferencias han sido complementadas con investigaciones empíricas, literatura específica y la revisión de los textos normativos vigentes. El instrumento de recolección de datos ha sido un cuestionario administrado durante los meses de marzo a octubre de 2017 en tres clínicas de fertilidad (una de Ciudad de Buenos Aires y dos de Madrid). Se puede inferir de los resultados que además de las motivaciones económicas para convertirse en donantes de óvulos, existen motivos altruistas. El altruismo entiéndase como construcción retórica de las clínicas y los consentimientos informados, destinada a eludir la mera consideración mercantil. Si bien un gran porcentaje de las mujeres encuestadas no desea conocer a las personas nacidas de sus óvulos, la mayoría considera que las personas deben conocer el modo en que han sido engendradas.

Conception narratives and third party reproduction in lesbian‐parented families in France

AbstractLesbian mothers and gay fathers cannot conceal the existence of a third party for reproduction purposes: a sperm donor, an egg cell donor or a surrogate mother, all of whom have contributed, without being a parent as such, to the birth of their child. This article focuses on what is said about these third parties and what the child is told about his or her conception, by lesbian mothers who used sperm donation, either through the means of MAP or with the help of a known donor, outside the medical field. The inquiry, both qualitative and quantitative, uses a sample of 118 female couples to whom a child was born in France between 2011 and 2013. Analysis shows that the mothers employ different terms by which they wish to be known by their child and by which to refer to a donor according to whether he is known or not. The study exposes not only a common structure to most of the conception stories but also certain variations revealing four types of narratives. The narratives produced by couples who have used a known donor insist less on the love story between the two mothers. The entirety of these narratives, however, offer the chance to reconcile a transgressive family model with society: to normalise and valorise the family set‐up.

218 Urine samples as a noninvasive source of induced pluripotent cells in the swine model

Noninvasive collection of cells used for generation of induced pluripotent stem cells (iPSC) generation would simplify its use for regenerative and reproductive purposes in veterinary medicine, although it is still unpublished in species other than humans. This study aimed to derive urine progenitor cells (UPCs) invitro from urine samples collected from swine, and then to reprogram them into iPSCs. For that, urine samples were collected from three females, and cells were isolated and cultured from each following the human UPCs protocol (Steichen et al. 2017 Curr. Protoc. Hum. Genet. 21, 7.1-21.7.22; https://doi.org/10.1002/cphg.26). Approximately 200mL of urine samples were collected in sterile flasks and centrifuged at 300×g; the pellet was washed in Dulbecco's PBS, resuspended, and cultured in 45% Dulbecco's modified Eagle's medium (DMEM)-high glucose, 5% fetal bovine serum (FBS), 50% Renal Epithelial Cell Growth basal medium (REBM) supplemented with 1% glutamine, 1% nonessential amino acids (NEAA), penicillin/streptomycin (P/S), and REGM supplements hEGF, insulin, hydrocortisone, GA-1000, FBS, transferrin, triiodothyronine, epinephrine (Lonza), and 10ngmL−1 basic fibroblast growth factor (bFGF). The UPCs first colonies were observed approximately 1 week after and resembled epithelial-like cells. At passage 2, one cell line was transduced with murine OCT4, SOX2, KLF4, and C-MYC cDNAs (OSKM) using a lentiviral vector. After 5 days, cells were plated onto mouse embryonic fibroblasts and cultured in knockout DMEM/F12, 20% knockout serum replacement, NEAA, L-glutamine, 2-mercaptoethanol, and P/S supplemented with 10ngmL−1 bFGF. Efficiency of reprogramming was 8.45%, measured by analysing the number of typical iPSC colonies relative to the transduced cells plated, after ~12 days. Three clonal lines (C1, C4, and C6) were maintained invitro and characterised regarding pluripotency markers for at least 30 passages. All three lines were positive for alkaline phosphatase activity in passages 15 and also 22. Immunocytochemistry analysis revealed that C6 (passage 22) was positive for the pluripotency genes OCT4 (1:100, SC), SOX2 (1:500, AB), SSEA1 (1:50, SC), TRA1- 81 (1:50, Millipore), and NANOG (1:100, AB), whereas C1 and C4 (passages 23 and 22, respectively) were positive only for OCT4, SOX2, and SSEA1. The expression of exogenous and endogenous pluripotency factors (OCT4, SOX2, and NANOG) was evaluated by qRT-PCR, comparing the three clonal lines at passages 16/17 and 21/22 and comparing different passages (10, 11, 14, 17, and 22) on C1 line. No statistical difference was observed between cell lines when compared in different passages, perhaps because of the great variation between lines. However, analysis of C1 line over time showed that pluripotency genes increased and exogenous vector expression decreased during early passages (±10 passages); however, after passage 17, OCT4 and NANOG decreased whereas SOX2 and exogenous vector expression increased. In conclusion, it was possible to reprogram UPCs into iPSCs and maintain them in culture for at least 30 passages; however, iPSCS were dependent on exogenous factors. These results represent the partial reprogramming of UPCs to iPSCs in animals for the first time, enabling the generation of invitro disease models using a noninvasive method. Financial support for this study was received from FAPESP (2019/02811-2, 2015/26818-5), CNPq (433133/2018-0), and CAPES.

The Israeli national population program of genetic carrier screening for reproductive purposes. How should it be continued?

The Israeli population genetic screening program for reproductive purposes, is a population-specific screening that includes all known, severe diseases and relatively frequent in a specific population (carrier frequency at or above 1:60 and/or disease frequency at or above 1 in 15,000 live births). The carrier screening program is free of charge and offers testing according to disease frequency in the different groups within the population.The extraordinary technical changes that occurred in the last decade as well as the changes in the type of marriages within the Israeli population necessitate a revision in the basis of the program.The screening should include instead of only the relatively frequent variants, all the variants that were reported among patients causing a severe disease for which the natural history is well known without regard of their frequency. The population-specific screening that determine which variants are included according to the origin of the couple should be abandoned for a general screening including either all the Jewish population or all the Israeli Arab population.

Reproductive biology of sailfin molly, Poecilia latipinna (Lesueur, 1821) in tambak Bosowa Kabupaten Maros

Sailfin molly, Poecilia latipinna is one of the alien ornamental fishes in Indonesia. This fish has been found in the aquaculture area in Maros Regency, south Sulawesi as a pest. The research aimed to identify species and observe aspects of reproductive biology of sailfin molly. The study was conducted in the Bosowa Isuma aquaculture area, in Maros Regency from November 2017 to April 2018. Fish was captured using a gillnet. The catches were separated for fish identifying and reproductive biology purposes. For the observation of reproductive biology, the samples were separated by sex according to external morphology. Measurement of total length and weight using calliper (mm) and analytical scales (g), respectively. The gonads preserved in the 4% formalin solution, used for determination of gonad developmental stages and fecundity. Identification results showed that the molly fish found in the ponds was Poecilia latipinna (Lesueur 1821). The length distribution of male and female fish was 26-76 and 31-66 mm with a mean of 51 and 46 mm, respectively. Overall, sex ratio and mature gonads between male and female fish were 1: 2 and 1:10; the growth patterns of male and female were negative allometrics and isometric, respectively. This fish is spawn throughout the year with the peak of spawning in January. The number of larvae to be born (larval fecundity) ranges from 12-111 individuals with an average of ± 32 larva/female.

Description and Outcomes of Current Clinical Techniques for Sperm Cryopreservation

Nowadays, sperm cryopreservation is strongly recommended in cases of malignancy. Moreover, the use of frozen testicular sperm in azoospermic patients prevents the need for repeated sperm retrieval and optimises scheduling between oocyte and sperm obtainment. Even though cryopreservation of human spermatozoa for assisted reproductive purposes is a widely implemented practice, none of the established freezing and vitrification techniques offer optimal cryosurvival results due to the dramatic impact of cryodamage on sperm cells. This comprehensive review describes the most commonly used sperm cryopreservation techniques in order to establish which of them minimise sperm cryodamage and offer better survival rates. Presently, it is not sufficiently demonstrated that sperm vitrification improves survival significantly more than freezing methods. Slow freezing offers the best survival results when compared to other freezing protocols, and owing to its technical advantages, can be considered as one of the preferred protocols to be easily implemented in assisted reproduction laboratories. Moreover, several studies have suggested that sperm preparation prior to cryopreservation can improve thawed sample quality. However, other authors have demonstrated that freezing the fresh sample and performing semen preparation after thawing gives better results in regard to total motile sperm count and motility. Regarding clinical results, it is well established that similar or even better reproductive outcomes are achieved using frozen testicular sperm in cases of azoospermia or anejaculation. Moreover, the use of frozen semen in cancer patients can help to achieve good fertilisation and pregnancy rates. Finally, the use of frozen sperm is not at all associated with worse post-natal development.

A pro-inflammatory CD8+ T-cell subset patrols the cervicovaginal tract.

The immune system of the cervicovaginal tract (CVT) must balance immunosurveillance and active immunity against pathogens with maintenance of tolerance to resident microbiota and to fetal and partner antigens for reproductive purposes. Thus, we predicted that CVT immunity is characterized by distinctive features compared to blood and other tissue compartments. Indeed, we found that CVT CD8+ T-cells had unique transcriptional profiles, particularly in their cytokine signature, compared to that reported for CD8+ T-cells in other tissue sites. Among these CVT CD8+ T-cells, we identified a CD69- CD103- subset that was characterized by reduced migration in response to tissue-exit signals and higher pro-inflammatory potential as compared to their blood counterpart. These inflammatory mucosal CD8+ T-cells (Tim) were increased in frequency in the CVT of individuals with chronic infection, pointing to a potential role in perpetuating inflammation. Our findings highlight the specialized nature of immunity within the CVT and identify Tim cells as potential therapeutic targets to tame tissue inflammation upon chronic infection.