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PURPOSE: To compare the DNA damage in granulosa cells (GCs) of women undergoing ovarian-stimulated cycles with four widely used recombinant human follicle-stimulating hormones (rhFSH) in in vitro fertilization (IVF) protocols (Corneumon®, Gonal-F®, Pergoveris® and Puregon®). METHODS: A randomized trial was carried out at a Mexican hospital. GCs were isolated from 18 women with infertility undergoing assisted reproductive techniques (ART). Four controlled ovarian stimulation (COS) protocols including Corneumon®, Gonal-F®, Pergoveris® or Puregon® were used. GCs DNA damage was assessed by the Comet assay. Two parameters were measured: comet tail length (CTL), and Olive tail moment (OTM, the percentage of DNA in the tail multiplied by the distance between the center of the tail and head). RESULTS: Use of the different hrFSH in COS caused variable and statistically significant levels of DNA damage in GCs of infertile women. CTL was similar in the Corneumon® and Pergoveris® groups (mean values of 48.73 and 55.18, respectively) and Corneumon® CTL was significantly lower compared to the Gonal-F® and Puregon® groups (mean values of 61.98 and 91.17, respectively). Mean OTM values were significantly lower in Corneumon® and Pergoveris® groups, compared to Gonal-F® and Puregon® groups (25.59, 27.35, 34.76, and 47.27, respectively). CONCLUSION: Use of Corneumon® and Pergoveris® in COS caused statistically significantly lower levels of DNA damage in GCs of infertile women undergoing ART, which could potentially correlate with better reproductive outcomes.

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Multiple effects of stress on health have been reported; however, reproductive alterations in oocytes and cumulus cells have not been fully described. In females, chronic stress has been shown to produce alterations in the estrous cycle, to decrease oocyte in vivo maturation, and to increase the percentage of abnormal oocytes. The aim of this study was to evaluate whether the oocytes from chronically stressed female rats could recover and mature in vitro by providing them with all the necessary culture conditions, as well as to evaluate the functionality of the GAP junctions, and the viability and DNA integrity of the cumulus cells, which are crucial for the complete maturation and development of the oocyte. For this, rats were stressed daily by cold water immersion (15 °C) during 15 min for 30 consecutive days. Corticosterone serum levels in rats increased as an indicator of stress. Chronic stress decreased the percentage of in vitro matured oocytes because the cumulus cells presented irreparable damage to their DNA that led to their death, being unable to establish bidirectional communication with the oocyte for its meiotic resumption through the GAP junctions, which were also damaged. These findings could partially explain an association between stress and infertility.

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RESUMEN La proteinosis alveolar pulmonar es una entidad clínica caracterizada por la acumula ción de material proteináceo, con alta riqueza en surfactante, mediado por una menor aclaración por parte de los macrófagos alveolares. En pacientes adultos, comúnmente se asocia a fenómenos autoinmunes que tienen como resultado una deficiencia del factor estimulante de colonias de granulocitos y macrófagos, lo que implica alteracio nes en la maduración y disfunción celular, lo que provoca disminución de la degrada ción del surfactante y su acumulación en el espacio alveolar. Su diagnóstico corres ponde a un reto para el clínico, sobre la base de los hallazgos en pruebas de función pulmonar, el patrón en "empedrado" (crazy paving) en la tomografía computarizada de tórax de alta resolución y que se confirma al obtener el material proteináceo en el lava do broncoalveolar. Dada su rareza, el tratamiento ideal permanece por ser elucidado y en la actualidad el pilar del tratamiento es el lavado pulmonar total. Reportamos un caso anecdótico de una paciente de 41 años con proteinosis alveolar pulmonar desde 2011, que ha requerido múltiples lavados pulmonares totales, con pobre respuesta a estos, persistencia de disnea y necesidad de oxígeno suplementario a pesar de realizar el procedimiento, pero con tendencia progresiva a la mejoría en los últimos 2 años. La técnica del lavado no está completamente estandarizada y su uso en Amé rica Latina es aún limitado, por lo que publicamos el protocolo utilizado en el Hospital Santa Clara de Bogotá, Colombia.

ABSTRACT Pulmonary alveolar proteinosis is a clinical entity characterized by the accumulation of proteinaceous material, rich in surfactant, mediated by reduced clearance by alveolar macrophages. In adult patients, it is commonly associated with autoimmune phenom ena resulting in a deficiency of the granulocyte-macrophage colony stimulating factor, which implies alterations in cell maturation and dysfunction, causing a decrease in surfactant degradation and its accumulation in the alveolar space. Its diagnosis poses a challenge to the clinician, based on the findings of pulmonary function tests and the crazy paving pattern of the high-resolution computed tomography of the chest, and is confirmed by obtaining the proteinaceous material in the bronchoalveolar la vage. Given its rarity, the ideal treatment remains to be elucidated, with whole lung lavage currently being the cornerstone of treatment. We report an anecdotal case of a 41-year-old female patient suffering from pulmonary alveolar proteinosis since 2011, who has required multiple whole lung lavages, with poor response to these, with persistent dyspnea and supplemental oxygen requirement even though she has performed the procedure, but with a progressive tendency towards improvement in the last 2 years. The lavage technique is not completely standardized and its use in Latin America is still limited, which is why we publish the protocol used in the Hospital Santa Clara of Bogotá, Colombia.

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El síndrome de Birt-Hogg-Dubé es una rara enfermedad autosómica dominante causada por la mutación patogénica del gen de la foliculina, que se expresa principal mente en tres órganos que incluyen el pulmón, la piel y el riñón, y produce quistes pulmonares, tumores renales y cutáneos. Desde el punto de vista respiratorio es poco sintomática, pero los quistes presentan alto riesgo de neumotórax, por lo que es imprescindible realizar una adecuada semiología radiológica de los quistes para un diagnóstico oportuno. Los tumores más importantes son los renales porque incluyen varios tipos de carcinomas renales; debido a esto requieren seguimiento estricto y, en muchos, casos cirugía. Presentamos dos casos de pacientes con este síndrome; uno confirmado por la mutación genética y el otro, por la confirmación histológica de fibrofoliculoma, ambos criterios mayores para el diagnóstico de esta enfermedad. Es fundamental el diagnóstico temprano de esta entidad de acuerdo con lo expuesto anteriormente, por lo que hacemos esta revisión con una amplia discusión sobre la afectación pulmonar, la semiología radiológica de los quistes y los criterios diagnósticos.

The Birt-Hogg-Dubé syndrome is a rare autosomal dominant disease caused by the pathogenic mutation of the folliculin gene, which is mainly expressed in three organs that include the lung, the skin and the kidney, and produces lung cysts, and renal and skin tumors. From the respiratory point of view, it doesn't have many symptoms, but cysts have high risk of pneumothorax, so it is indispensable to carry out the correct radiological semiology of the cysts for a timely diagnosis. The most important tumors are the renal, because they include several types of renal carcinomas; that is why they require strict follow-up and, in many cases, surgery. We present two cases of patients with this syndrome: one confirmed by the genetic mutation, and the other one by the histological confirmation of fibrofolliculoma, both major criteria for the diagnosis of this disease. The early diagnosis of this entity is of fundamental importance, according to what has been previously presented, so we conduct this review with a broad discus sion about lung involvement, the radiological semiology of the cysts, and diagnostic criteria.

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Phytoestrogens are considered to be endocrine disruptors, since they can alter the endocrine system, thus disturbing many reproductive events. The intake of diets containing a high content of phytoestrogens has increased worldwide in human populations and in domestic animals. Phytoestrogens in maternal blood can pass through the placenta to the fetus in high amounts and can have long-term organizational effects. Mesquite (Prosopis sp) is a leguminous plant widely used to feed several livestock species, and is also used in the human diet. In this study we assessed the effects of exposure to mesquite pod extract during the periconception and pregnancy periods on the reproduction of male and female descendants. The females of three experimental groups received one of the following treatments: 1) vehicle injection; 2) mesquite pod extract or 3) the isoflavone daidzein during the periconception and pregnancy periods. Estrous cyclicity, sexual behavior and hormones, as well as uterine and vaginal epithelia were evaluated in the female descendants. In the males, sexual behavior and hormones, apoptosis in testicular cells and sperm quality were evaluated. In females the following was observed: alterations in estrous cycles, decreased sexual behavior, estradiol and progesterone levels, increased uterine and vaginal epithelia. In males, we observed a decrease in sexual behavior, testosterone and sperm quality, and apoptosis increased in testicular cells. All these effects were similar to those caused by daidzein. These results indicate that prenatal exposure to mesquite pod extract or daidzein, administered to females before and during pregnancy, can disrupt normal organizational-activational programming of reproductive physiology in female and male descendants.

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The reactive oxygen species (ROS) play an important role in various aspects of male reproductive function, for spermatozoa to acquire the ability to fertilize. However, the increase in ROS generation, both due to internal and external factors, can induce oxidative stress, causing alterations in the structure and function of phospholipids and proteins. In the nucleus, ROS attack DNA, causing its fragmentation and activation of apoptosis, thus altering gene and protein expression. Accumulating evidence also reveals that endogenously produced ROS can act as second messengers in regulating cell signalling pathways and in the transduction of signals that are responsible for regulating spermatogonia self-renewal and proliferation. In the epididymis, they actively participate in the formation of disulphide bridges required for the final condensation of chromatin, as well as in the phosphorylation and dephosphorylation of proteins contained in the fibrous sheath of the flagellum, stimulating the activation of progressive motility in epididymal spermatozoa. In this review, the role of small amounts of ROS during spermatogenesis and epididymal sperm maturation was discussed.

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BACKGROUND: The emergence of assisted reproductive technology (ART) in humans has been an important tool for the treatment of infertility. The number of treatments performed in Latin America has been increasing, and Mexico is the third country with the most assisted reproduction cycles performed in the region. However, Mexico lacks a national regulation for assisted reproduction. Therefore, it is necessary to implement regulations that allow for a safe clinical practice based on ethics which can be available to any social group. MAIN BODY: The aim of this review was to examine the existing legislation that regulates human assisted reproduction practices in Mexico, but also to examine the legal analysis of the policies, laws, and regulations in effect in some countries in Latin America, North America, and Europe. For this, seven databases were consulted, and 34 articles from 2004 to 2021 referring to the practice of ART within the legal framework and the anthropological analysis that this entails were analyzed. Eight documents were also consulted such as the Mexican General Health Law of the Official Journal of the Federation (February 7, 1984) with its last published reform (DOF 01-06-2021). And three official agency websites were also consulted. No specific legislation was found for human assisted reproduction practices in Mexico; however, assisted reproduction clinics are ruled under some agreements implemented by national organizations such as the Mexican Association of Reproductive Medicine and, at the Latin America level, the Latin America Network of Assisted Reproduction (abbreviated REDLARA in Spanish); in addition, the practice of ART is considered, although not explicitly, in the General Health Law. CONCLUSION: In Mexico, there is no legal regulation in charge of assisted reproduction practices, which is why there is an urgent need to establish human assisted reproduction laws without incurring discriminatory and unconstitutional acts, and at the same time, be in accordance with scientific advances. This will allow a considerable reduction in the violation of human rights.

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BACKGROUND: The evaluation of the DNA damage generated in cumulus cells after mature cumulus-oocyte complexes vitrification can be considered as an indicator of oocyte quality since these cells play important roles in oocyte developmental competence. Therefore, the aim of this study was to determine if matured cumulus-oocyte complexes exposure to cryoprotectants (CPAs) or vitrification affects oocytes and cumulus cells viability, but also if DNA damage is generated in cumulus cells, affecting fertilization and embryo development. RESULTS: The DNA damage in cumulus cells was measured using the alkaline comet assay and expressed as Comet Tail Length (CTL) and Olive Tail Moment (OTM). Results demonstrate that oocyte exposure to CPAs or vitrification reduced oocyte (75.5 ± 3.69%, Toxicity; 66.7 ± 4.57%, Vitrification) and cumulus cells viability (32.7 ± 5.85%, Toxicity; 7.7 ± 2.21%, Vitrification) compared to control (95.5 ± 4.04%, oocytes; 89 ± 4.24%, cumulus cells). Also, significantly higher DNA damage expressed as OTM was generated in the cumulus cells after exposure to CPAs and vitrification (39 ± 17.41, 33.6 ± 16.69, respectively) compared to control (7.4 ± 4.22). In addition, fertilization and embryo development rates also decreased after exposure to CPAs (35.3 ± 16.65%, 22.6 ± 3.05%, respectively) and vitrification (32.3 ± 9.29%, 20 ± 1%, respectively). It was also found that fertilization and embryo development rates in granulose-intact oocytes were significantly higher compared to denuded oocytes in the control groups. However, a decline in embryo development to the blastocyst stage was observed after CPAs exposure (1.66 ± 0.57%) or vitrification (2 ± 1%) compared to control (22.3 ± 2.51%). This could be attributed to the reduction in both cell types viability, and the generation of DNA damage in the cumulus cells. CONCLUSION: This study demonstrates that oocyte exposure to CPAs or vitrification reduced viability in oocytes and cumulus cells, and generated DNA damage in the cumulus cells, affecting fertilization and embryo development rates. These findings will allow to understand some of the mechanisms of oocyte damage after vitrification that compromise their developmental capacity, as well as the search for new vitrification strategies to increase fertilization and embryo development rates by preserving the integrity of the cumulus cells.

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BACKGROUND: Stress has been considered as one of the causes of decreased reproductive function in women. However, direct evidence of the effect of chronic stress on oocytes depending on estrous cycle phases is limited. OBJECTIVE: The present study aimed to evaluate the impact of chronic stress on the viability, integrity, and maturation of rat oocytes depending on estrous cycle phases, specifically proestrus, estrus, and diestrus. METHODS: For this purpose, adult female rats were stressed daily by cold water immersion (15 °C) for 30 consecutive days. RESULTS: In chronically stressed female rats, irregular estrous cyclicity, increased corticosterone levels, decreased oocyte viability, and an increased percentage of abnormal oocytes were obtained in all the estrous cycle phases, resulting in reduced oocyte maturation during proestrus. CONCLUSION: Oocyte maturation disturbed by chronic stress is a crucial factor by which chronic stress disrupts female reproduction.

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Vitrification is mainly used to cryopreserve female gametes. This technique allows maintaining cell viability, functionality, and developmental potential at low temperatures into liquid nitrogen at -196°C. For this, the addition of cryoprotectant agents, which are substances that provide cell protection during cooling and warming, is required. However, they have been reported to be toxic, reducing oocyte viability, maturation, fertilization, and embryo development, possibly by altering cell cytoskeleton structure and chromatin. Previous studies have evaluated the effects of vitrification in the germinal vesicle, metaphase II oocytes, zygotes, and blastocysts, but the knowledge of its impact on their further embryo development is limited. Other studies have evaluated the role of actin microfilaments and chromatin, based on the fertilization and embryo development rates obtained, but not the direct evaluation of these structures in embryos produced from vitrified immature oocytes. Therefore, this study was designed to evaluate how the vitrification of porcine immature oocytes affects early embryo development by the evaluation of actin microfilament distribution and chromatin integrity. Results demonstrate that the damage generated by the vitrification of immature oocytes affects viability, maturation, and the distribution of actin microfilaments and chromatin integrity, observed in early embryos. Therefore, it is suggested that vitrification could affect oocyte repair mechanisms in those structures, being one of the mechanisms that explain the low embryo development rates after vitrification.