Effect of gamete aging on fertilization success of the sea urchin Arbacia dufresnii (Arbacioida: Arbaciidae) / Efecto del envejecimiento de los gametos sobre el éxito en la fecundación del erizo de mar Arbacia dufresnii (Arbacioida: Arbaciidae)

Abstract Introduction: Short-term gametes storage is an inexpensive and simple technique that allows the use of the same batch of eggs or sperm at different times, maximizing the application of research protocols and the use of gametes in production. Arbacia dufresnii is a sea urchin species with proven aquaculture potential and already used in the nutraceutical industry. Aging of its gametes is unknown and is a needed information to scale up the production. Objective: Determine the effect of male and female gamete aging on the fertilization success of Arbacia dufresnii. This will allow optimizing the use of gametes after collection decoupling spawning from fertilization. Methods: A. dufresnii individuals were induced to spawn and gametes were kept at 12 ± 1 °C throughout each bioassay. Sperm was separated into two treatments: activated sperm in seawater (AS), and dry sperm (DS). Two bioassays were made: Bioassay 1 evaluated the effect of time on fertility by performing fertilization tests at 0 h, 24 h, 48 h, 72 h, and 96 h after spawning. Bioassay 2 evaluated the contribution of each type of aged gamete on fertility, combining aged gametes (96 h) with fresh gametes (0 h). Results: Bioassay 1: the fertilization success obtained by combining eggs (E) with AS or DS presented important differences. While the fertilization success remained acceptable (greater than 50 %) for up to 72 h using ExDS, it only remained acceptable for up to 48 h using ExAS. Bioassay 2: acceptable fertilization success was found by combining aged E (96 h) with fresh sperm, or aged DS (96 h) with fresh E, but not using aged AS with fresh E. Conclusions: The findings of this work show that fertilization success in A. dufresnii gametes remains relatively unchanged for up to 48 h after spawning when combining ExAS, and for up to 72 h when combining ExDS. However, when combining aged E or aged DS with a fresh gamete, post-collection fertilization can be extended up to 96 h. In this work, the first steps have been taken to understand the conservation time of A. dufresnii gametes with minimum intervention.

Resumen Introducción: El almacenamiento de gametos a corto plazo es una técnica económica y sencilla que permite utilizar el mismo lote de óvulos o espermatozoides en diferentes momentos, maximizando la aplicación de protocolos de investigación y el uso de gametos en la producción. Arbacia dufresnii es una especie con probado potencial acuícola como fuente de gametos para la industria nutracéutica. Sin embargo, se desconoce el envejecimiento de sus gametos y es una información necesaria para escalar la producción. Objetivo: Determinar el efecto del envejecimiento de los gametos masculinos y femeninos en el éxito de la fecundación de Arbacia dufresnii con el fin de optimizar el aprovechamiento de los gametos después de la recolecta desincronizando el desove de la fecundación. Métodos: Se indujo el desove de individuos de A. dufresnii y los gametos se mantuvieron a 12 ± 1 °C durante cada bioensayo. El esperma se separó en dos tratamientos: esperma activado en agua de mar (AS) y esperma seco (DS). Se realizaron dos bioensayos: El Bioensayo 1 evaluó el efecto del tiempo sobre la fertilidad realizando pruebas de fecundación a las 0 h, 24 h, 48 h, 72 h y 96 h después del desove. El bioensayo 2 evaluó la contribución de cada tipo de gameta envejecida (96 h) sobre la fertilidad, combinando gametos envejecidas (96 h) con gametos frescas (0 h). Resultados: Bioensayo 1: el éxito de fecundación obtenido combinando huevos (E) con AS o DS presentó diferencias importantes. Si bien el éxito de la fecundación se mantuvo aceptable (más del 50 %) durante un máximo de 72 h con ExDS, solo permaneció aceptable hasta 48 h con ExAS. Bioensayo 2: se encontró un éxito de fecundación aceptable combinando E envejecidos (96 h) con esperma fresco, o DS envejecido (96 h) con E fresco (0 h), pero no usando AS envejecido con E fresco (0 h). Conclusiones: Los hallazgos de este trabajo muestran que el éxito de la fecundación en los gametos de A. dufresnii permanece relativamente sin cambios hasta 48 h después del desove cuando se combina ExAS, y hasta 72 h cuando se combina ExDS. Sin embargo, cuando se combina E envejecido o DS envejecido con un gameto fresco, el tiempo entre la recolección y la fecundación puede extenderse hasta 96 h. En este trabajo se han dado los primeros pasos para entender el tiempo de conservación de los gametos de A. dufresnii con mínima intervención.

Therapeutic effects of melatonin in long-term exposure to 2100MHz radiofrequency radiation on rat sperm characteristics / Efectos terapéuticos de la melatonina en la exposición a largo plazo a la radiación de radiofrecuencia de 2100MHz en las características del esperma de rata

Introduction: Radiofrequency electromagnetic fields (RF-EMFs) are one of the risk factors for male reproductive health and melatonin can be an ideal candidate for therapeutic development against RF-induced male fertility problems due to its antioxidant properties. The possible therapeutic role of melatonin in the destructive effects of 2100MHz RF radiation on rat sperm characteristics is investigated in the present study. Methods: Wistar albino rats were divided into four groups and the experiment continued for ninety consecutive days; Control, Melatonin (10mg/kg, subcutaneously), RF (2100MHz, thirty minutes per day, whole-body), and RF+Melatonin groups. Left caudal epididymis and ductus deferens tissues were placed in sperm wash solution (at 37°C) and dissected. The sperms were counted and stained. Measurements of the perinuclear ring of the manchette and posterior portion of the nucleus (ARC) were performed and the sperms were examined at an ultrastructural level. All of the parameters were evaluated statistically. Results: The percentages of abnormal sperm morphology were significantly increased with RF exposure, while the total sperm count was significantly decreased. RF exposure also showed harmful effects on acrosome, axoneme, mitochondrial sheath, and outer dense fibers at the ultrastructural level. The number of total sperms, sperms with normal morphology increased, and ultrastructural appearance returned to normal by melatonin administration. Discussion: The data showed that melatonin may be a beneficial therapeutic agent for long-term exposure of 2100MHz RF radiation-related reproductive impairments. (AU)

Introducción: Los campos electromagnéticos de radiofrecuencia (RF-EMF) son uno de los factores de riesgo para la salud reproductiva masculina y la melatonina puede ser un candidato ideal para el desarrollo terapéutico contra los problemas de fertilidad masculina inducidos por RF debido a sus propiedades antioxidantes. En el presente estudio se investiga el posible papel terapéutico de la melatonina en los efectos destructivos de la radiación RF de 2100MHz en las características del esperma de rata. Métodos: Se dividieron ratas albinas Wistar en 4 grupos y se continuó el experimento durante 90 días consecutivos: grupos control, melatonina (10mg/kg, por vía subcutánea), RF (2100MHz, 30min por día, cuerpo entero) y RF+melatonina. Los tejidos del epidídimo caudal izquierdo y del conducto deferente se colocaron en una solución de lavado de esperma (a 37°C) y se diseccionaron. Los espermatozoides fueron contados y teñidos. Se realizaron mediciones del anillo perinuclear del manchette y de la porción posterior del núcleo (ARC) y se examinaron los espermatozoides a nivel ultraestructural. Todos los parámetros fueron evaluados estadísticamente. Resultados: Los porcentajes de morfología anormal de los espermatozoides aumentaron significativamente con la exposición a RF, mientras que el recuento total de espermatozoides disminuyó significativamente. La exposición a RF también mostró efectos nocivos en el acrosoma, el axonema, la vaina mitocondrial y las fibras densas externas a nivel ultraestructural. El número total de espermatozoides, los espermatozoides con morfología normal aumentaron y la apariencia ultraestructural volvió a la normalidad mediante la administración de melatonina. Discusión: Los datos mostraron que la melatonina puede ser un agente terapéutico beneficioso para la exposición a largo plazo de las deficiencias reproductivas relacionadas con la radiación de RF de 2100MHz. (AU)

Effect of icariin on the transformation efficiency of induced pluripotent stem cells into sperm cells in vitro / Estudio de los efectos de icariina en la eficiencia de la transformación de espermatozoides extrovertidos inducidos en cuerpos multicompetentes

Objective: To investigate the effect of icariin on the transformation efficiency of germ cell-like cells from mouse induced pluripotent stem cells into sperm cells in vitro. Methods: Firstly, mouse induced pluripotent stem cells were induced and cultured to transform into germ cell-like cells, and the primordial germ cell-like cells were identified by Western blot and RT-PCR. Then, different concentrations of icariin (0.1μg/mL, 1μg/mL, 10μg/mL and 100μg/mL) were added into the culture medium, and the obtained primitive germ cell-like cells were cultured, Western blot and RT-PCR were used to identify the obtained sperm cells, the transformation efficiency was compared. Results: The primordium germ cell-like cells obtained from mouse induced pluripotent stem cells in vitro specially expressed Oct-4 protein, C-kit protein, Mvh mRNA, Fragilis mRNA and Stella mRNA. The sperm cells were specially expressed VASA, SCP3 and γH2AX proteins. RT-PCR showed that the sperm cells were specially expressed Ddx4, Tp2 and Prm1 mRNA. Compared with the control group, the expression level of VASA protein (1.744±0.283, 2.882±0.373, 6.489±0.460), SCP3 protein (2.250±0.306, 7.058±0.521, 8.654±0.804), γH2AX protein (4.304±0.433, 5.713±0.339, 9.268±0.545), Ddx4 mRNA (1.374±0.145, 2.846±0.194, 4.021±0.154), Tp2 mRNA (1.358±0.130, 3.623±0.326, 5.811±0.390) and Prm1 mRNA (1.326±0.162, 3.487±0.237, 4.666±0.307) in 0.1μg/mL, 1μg/mL, 10μg/mL icariin experimental groups were all lower than that of VASA protein (10.560±0.413), SCP3 protein (13.804±0.642), γH2AX protein (11.874±0.464), Ddx4 mRNA (6.4005±0.361), Tp2 mRNA (7.314±0.256) and Prm1 mRNA (7.334±0.390) in 100μg/mL icariin experimental group. (AU)

Objetivo: Investigar el efecto de icariina en la eficiencia de la conversión in vitro inducida en espermatozoides de cultivos de células germinativas derivadas de la transformación de células madre pluripotentes inducidas de ratón. Métodos: Primero se indujeron y cultivaron células madre pluripotentes inducidas de ratón para transformarlas en células similares a las células germinales, y las células similares a las células germinales primordiales se identificaron mediante Western blot y RT-PCR. A continuación, se añadieron diferentes concentraciones de icariina (0,1μg/mL, 1μg/mL, 10μg/mL and 100μg/mL) al medio de cultivo, y se cultivaron las células primitivas similares a células germinales obtenidas, se utilizaron Western blot y RT-PCR para identificar las células espermáticas obtenidas, y se comparó la eficacia de la transformación. Resultados: Las células germinales primitivas obtenidas in vitro a partir de células madre pluripotentes inducidas de ratón expresaron especialmente la proteína Oct-4, la proteína C-kit, el ARNm de Mvh, el ARNm de Fragilis y el ARNm de Stella. Los espermatozoides expresaban especialmente las proteínas VASA, SCP3 y γH2AX. La RT-PCR mostró que los espermatozoides expresaban especialmente los ARNm Ddx4, Tp2 y Prm1. En comparación con el grupo de control, el nivel de expresión de la proteína VASA (1,744±0,283; 2,882±0,373; 6,489±0,460), la proteína SCP3 (2,250±0,306; 7,058± 0,521; 8,654±0,804), proteína γH2AX (4,304±0,433; 5,713±0,339; 9,268±0,545), ARNm Ddx4 (1,374±0,145; 2,846±0,194; 4,021±0,154), ARNm Tp2 (1,358±0,130; 3,623±0,326; 5,811±0,390) y ARNm Prm1 (1,326±0,162; 3,487±0,237; 4,666±0,307) en grupos experimentales de 0,1μg/mL, 1μg/mL, 10μg/mL de icariina fueron todos más bajos que los de la proteína VASA (10,560±0,413), proteína SCP3 (13,804±0,642), proteína γH2AX (11,874±0,464), ARNm Ddx4 (6,4005±0,361), ARNm Tp2 (7,314±0,256) y ARNm Prm1 (7,334±0,390) en 100μg/mL icariina grupo experimental. (AU)

Postnatal Exposure to Exogenous progesterone Exacerbates the Prenatally-Induced Abnormal Sperms Production and Function in Rats / La exposición Posnatal a la Progesterona Exógena Exacerba la Producción y Función Anormales de Espermatozoides Inducidas Prenatalmente en Ratas

SUMMARY: This study aimed at clarifying the impact of long-term prenatal and postnatal exposure to exogenous progesterone on sperm production and function, relative sex organs weights, and the levels of the relevant hormones in rats. Sixty male Wistar rats were included and classified into three groups (n=20 in each). A test I group had mature rats born to dams treated with progesterone prenatally. A test II group included rats exposed to progesterone during prenatal as well as postnatal periods, and a control group had rats treated with a placebo (olive oil). The test groups revealed a significant reduction in sperm count, motility, and viability with higher abnormal forms than the control group (P< 0.05). Similarly, the test groups revealed significantly lower serum testosterone and higher FSH and LH levels (P< 0.001). Interestingly, the test II group showed pronounced sperm abnormalities, an alarming decrease in sperm viability and motility, and a significant accretion in the relative testicular weight compared to the test I group (p <0.001). Long-term (prenatal and early postnatal) treatment with synthetic progesterone hurts sperm quantity and quality, adversely affecting future male fertility.

Este estudio tuvo como objetivo aclarar el impacto de la exposición prenatal y posnatal a largo plazo a la progesterona exógena en la producción y función de los espermatozoides, el peso relativo de los órganos sexuales y los niveles de las hormonas relevantes en ratas. Sesenta ratas macho Wistar fueron incluidas y clasificadas en tres grupos (n=20 en cada uno). Un grupo de prueba I tenía ratas maduras nacidas de madres tratadas con progesterona prenatalmente. Un grupo de prueba II incluyó ratas expuestas a progesterona durante los períodos prenatal y posnatal, y un grupo de control tenía ratas tratadas con un placebo (aceite de oliva). Los grupos de prueba revelaron una reducción significativa en el recuento, la motilidad y la viabilidad de los espermatozoides con formas anormales más altas que el grupo de control (P < 0,05). De manera similar, los grupos de prueba revelaron niveles significativamente más bajos de testosterona sérica y niveles más altos de FSH y LH (P < 0.001). Curiosamente, el grupo de prueba II mostró anormalidades espermáticas pronunciadas, una disminución alarmante en la viabilidad y motilidad de los espermatozoides y una acumulación significativa en el peso testicular relativo en comparación con el grupo de prueba I (p <0.001). El tratamiento a largo plazo (prenatal y posnatal temprano) con progesterona sintética daña la cantidad y la calidad del esperma, lo que afecta negativamente la futura fertilidad masculina.

Effect of icariin on the transformation efficiency of induced pluripotent stem cells into sperm cells in vitro.

OBJECTIVE: To investigate the effect of icariin on the transformation efficiency of germ cell-like cells from mouse induced pluripotent stem cells into sperm cells in vitro. METHODS: Firstly, mouse induced pluripotent stem cells were induced and cultured to transform into germ cell-like cells, and the primordial germ cell-like cells were identified by Western blot and RT-PCR. Then, different concentrations of icariin (0.1µg/mL, 1µg/mL, 10µg/mL and 100µg/mL) were added into the culture medium, and the obtained primitive germ cell-like cells were cultured, Western blot and RT-PCR were used to identify the obtained sperm cells, the transformation efficiency was compared. RESULTS: The primordium germ cell-like cells obtained from mouse induced pluripotent stem cells in vitro specially expressed Oct-4 protein, C-kit protein, Mvh mRNA, Fragilis mRNA and Stella mRNA. The sperm cells were specially expressed VASA, SCP3 and γH2AX proteins. RT-PCR showed that the sperm cells were specially expressed Ddx4, Tp2 and Prm1 mRNA. Compared with the control group, the expression level of VASA protein (1.744±0.283, 2.882±0.373, 6.489±0.460), SCP3 protein (2.250±0.306, 7.058±0.521, 8.654±0.804), γH2AX protein (4.304±0.433, 5.713±0.339, 9.268±0.545), Ddx4 mRNA (1.374±0.145, 2.846±0.194, 4.021±0.154), Tp2 mRNA (1.358±0.130, 3.623±0.326, 5.811±0.390) and Prm1 mRNA (1.326±0.162, 3.487±0.237, 4.666±0.307) in 0.1µg/mL, 1µg/mL, 10µg/mL icariin experimental groups were all lower than that of VASA protein (10.560±0.413), SCP3 protein (13.804±0.642), γH2AX protein (11.874±0.464), Ddx4 mRNA (6.4005±0.361), Tp2 mRNA (7.314±0.256) and Prm1 mRNA (7.334±0.390) in 100µg/mL icariin experimental group. CONCLUSIONS: Icariin can promote the transformation of mouse induced pluripotent stem cells into sperm cells in vitro, and it is concentration-dependent manner in a certain concentration range.

Therapeutic effects of melatonin in long-term exposure to 2100MHz radiofrequency radiation on rat sperm characteristics.

INTRODUCTION: Radiofrequency electromagnetic fields (RF-EMFs) are one of the risk factors for male reproductive health and melatonin can be an ideal candidate for therapeutic development against RF-induced male fertility problems due to its antioxidant properties. The possible therapeutic role of melatonin in the destructive effects of 2100MHz RF radiation on rat sperm characteristics is investigated in the present study. METHODS: Wistar albino rats were divided into four groups and the experiment continued for ninety consecutive days; Control, Melatonin (10mg/kg, subcutaneously), RF (2100MHz, thirty minutes per day, whole-body), and RF+Melatonin groups. Left caudal epididymis and ductus deferens tissues were placed in sperm wash solution (at 37°C) and dissected. The sperms were counted and stained. Measurements of the perinuclear ring of the manchette and posterior portion of the nucleus (ARC) were performed and the sperms were examined at an ultrastructural level. All of the parameters were evaluated statistically. RESULTS: The percentages of abnormal sperm morphology were significantly increased with RF exposure, while the total sperm count was significantly decreased. RF exposure also showed harmful effects on acrosome, axoneme, mitochondrial sheath, and outer dense fibers at the ultrastructural level. The number of total sperms, sperms with normal morphology increased, and ultrastructural appearance returned to normal by melatonin administration. DISCUSSION: The data showed that melatonin may be a beneficial therapeutic agent for long-term exposure of 2100MHz RF radiation-related reproductive impairments.

Effects of photobiomodulation therapy on human sperm function / Efectos de la terapia de fotobiomodulación en la función del esperma humano

Introduction: Sperm motility is a crucial factor in male infertility and it depends on mitochondrial tail movements. Photobiomodulation light therapy allows the cells to produce their energy through activation of the mitochondria. The aim of the present study was to examine the impact of photobiomodulation on sperm motility in astenozoospermic individuals. Materials and methods: Following semen analyses of 20 astenozoospermic individuals, collected semen samples were centrifuged. Pellet was obtained and homogenized through mixing with culture media in 1:1 ratio. Each semen samples were divided into 3 groups. In the first group, control samples were not exposed to laser irradiation. The Group 2 and Group 3 were exposed to 650nm wavelength of photobiomodulation from 10cm distance in dark environment via a 36cm2 aperture sizer with 200mW output power for 30 and 60min duration, respectively. Sperm motilities were evaluated and chromatin condensation of sperms was determined. Results: Sperm motilities were significantly increased in photobiomodulation groups compared with the controls. Sperm motilities tended to be different between the 30 and 60min red light exposure groups; however, it was not statistically significant. When the motility grades were compared, no significant difference was observed in non-progressive motility sperms. While immotile sperms decreased significantly in the photobiomodulation groups compared to the control group, progressive sperms increased. (AU)

Introducción: La motilidad espermática es un factor crucial en la infertilidad masculina y depende de los movimientos de la cola mitocondrial. La fototerapia de fotobiomodulación permite que las células produzcan su energía a través de la activación de las mitocondrias. El objetivo del presente estudio fue examinar el impacto de la fotobiomodulación en la motilidad de los espermatozoides en individuos astenozoospérmicos. Materiales y métodos: Luego de los análisis de semen de 20 individuos astenozoospérmicos, se centrifugaron las muestras de semen recolectadas. Se obtuvo el sedimento y se homogeneizó mezclándolo con medios de cultivo en una proporción de 1:1. Cada muestra de semen se dividió en 3 grupos. En el primer grupo, las muestras de control no se expusieron a la irradiación láser. El grupo 2 y el grupo 3 fueron expuestos a una longitud de onda de 650nm de fotobiomodulación desde una distancia de 10cm en un ambiente oscuro a través de un medidor de apertura de 36cm2 con una potencia de salida de 200mW durante 30 y 60min de duración, respectivamente. Se evaluaron las motilidades de los espermatozoides y se determinó el tamaño de la cromatina de los espermatozoides. Resultados: La motilidad de los espermatozoides aumentó significativamente en los grupos de fotobiomodulación en comparación con los controles. La motilidad de los espermatozoides tendió a ser diferente entre los grupos de exposición a la luz roja de 30 y 60min; sin embargo, no fue estadísticamente significativo. Cuando se compararon los grados de motilidad, no se observaron diferencias significativas en los espermatozoides de motilidad no progresiva. Mientras que los espermatozoides inmóviles disminuyeron significativamente en los grupos de fotobiomodulación en comparación con el grupo de control, los espermatozoides progresivos aumentaron. (AU)

Effects of photobiomodulation therapy on human sperm function.

INTRODUCTION: Sperm motility is a crucial factor in male infertility and it depends on mitochondrial tail movements. Photobiomodulation light therapy allows the cells to produce their energy through activation of the mitochondria. The aim of the present study was to examine the impact of photobiomodulation on sperm motility in astenozoospermic individuals. MATERIALS AND METHODS: Following semen analyses of 20 astenozoospermic individuals, collected semen samples were centrifuged. Pellet was obtained and homogenized through mixing with culture media in 1:1 ratio. Each semen samples were divided into 3 groups. In the first group, control samples were not exposed to laser irradiation. The Group 2 and Group 3 were exposed to 650nm wavelength of photobiomodulation from 10cm distance in dark environment via a 36cm2 aperture sizer with 200mW output power for 30 and 60min duration, respectively. Sperm motilities were evaluated and chromatin condensation of sperms was determined. RESULTS: Sperm motilities were significantly increased in photobiomodulation groups compared with the controls. Sperm motilities tended to be different between the 30 and 60min red light exposure groups; however, it was not statistically significant. When the motility grades were compared, no significant difference was observed in non-progressive motility sperms. While immotile sperms decreased significantly in the photobiomodulation groups compared to the control group, progressive sperms increased. CONCLUSIONS: The results of the present study demonstrated that the photobiomodulation is an efficient method to increase the sperm motility of astenozoospermic individuals independent of the duration of exposure.

Is there a relationship between infertility and fertilin β protein distribution? / ¿Existe relación entre la infertilidad y la distribución de la proteína fertilin β?

Introduction: Fertilin β is a sperm surface protein that can mediate sperm-egg membrane interaction. This study was conducted to determine whether the expression of fertilin β after intrauterine insemination (IUI) in donors with normal parameters after standard semen analysis is related to low success rate or failure of fertilization. Methods: We examined the sperm of 30 male donors who have normal as controls, oligozoospermia, and unexplained infertility as the clinically indication for IUI. Fertilin β has been labeled with the ADAM2 antibody by indirect immunofluorescence (IF) assay. To evaluate the reproducibility of the test, we selected four sperm samples scale of 0 to +++ according to the distribution of fluorescence label. Results: The results were highly correlated with the corrected total cell fluorescence (CTCF) (Rp=0.9972, P<0.05). We suggest that the relationship between infertility and fertilin β may be due to the distribution of this protein on the sperm surface. Male partners of couples with unexplained infertility showed a low distribution of fertilin β by a decrease of the fluorescence signal in the IF labeling (scale of +++ by 7.4±10.32%, P<0.0001, ±SD). Discussion: Abnormal fertilin β function may be a potential mechanism that could lead to fertilization failure. (AU)

Introducción: La fertilinβ es una proteína de la superficie del esperma que puede mediar entre la interacción del espermatozoide y la membrana del óvulo. Este estudio se realizó para determinar si la expresión de fertilinβ después de la inseminación intrauterina (IIU) en donantes con parámetros normales después del análisis estándar de semen está relacionada con una baja tasa de éxito o fracaso de la fertilización. Métodos: Examinamos los espermatozoides de 30 donantes masculinos que tenían controles normales, oligozoospermia e infertilidad inexplicable como indicación clínica de IIU. La fertilinβ ha sido etiquetada con el anticuerpo ADAM2 mediante un ensayo de inmunofluorescencia indirecta (IF). Para evaluar la reproducibilidad de la prueba, seleccionamos cuatro muestras de esperma en una escala de 0 a +++ según la distribución de la etiqueta de fluorescencia. Resultados: Los resultados estuvieron altamente correlacionados con la fluorescencia celular total corregida (Rp=0,9972, p<0,05). Sugerimos que la relación entre la infertilidad y la fertilinβ puede deberse a la distribución de esta proteína en la superficie del esperma. Los compañeros masculinos de parejas con infertilidad inexplicable mostraron una baja distribución de fertilinβ por una disminución de la señal de fluorescencia en el etiquetado IF (escala de +++ en 7,4 ±10,32%, p<0,0001, ±DE). Conclusión: La función anormal de la fertilinβ puede ser un mecanismo potencial que podría conducir al fracaso de la fertilización. (AU)

Cyperus esculentus L. protects testis and sperm morphology of hyperglycaemic rats / Cyperus esculentus L. protege la morfología de los testículos y los espermatozoides de ratas hiperglicémicas

Cyperus esculentus L. (tiger nut) is a tuberous plant that promotes and protects reproductive functions, which are usually hampered in diabetics. The present study investigated the effect of Cyperus esculentus tuber extract (CETE) on testicular histology and sperm viability of alloxan-induced hyperglycaemic Wistar rats. Twenty-five adult male Wistar rats weighing 150-200g and grouped into five (n=5): Group 1, the control, administered tap water (20mL/kg), while groups 2-5 were administered a single intraperitoneal dose (120mg/kg b.w.) of alloxan, and each further received orally tap water (20mL/kg), CETE (100mg/kg), CETE (500 mg/kg) and metformin (500 mg/kg), respectively for 21 days. The animals were sacrificed, their sperm collected for analysis, while the testes were harvested, and processed for histology. Results showed significantly increased (p<0.05) blood glucose and testosterone, and significantly decreased (p<0.05) sperm pH, motility, count, morphology and density, as well as disruptions and hypertrophy of the spermatogenic and Sertoli cells of the hyperglycaemic group. There were significant (p<0.05) blood glucose decline, while the sperm parameters and testicular weight improved with normal testicular histology in the 100 mg/kg CETE, 500 mg/kg CETE, and metformin-treated groups compared to the control and hyperglycaemic group. Treatment with CETE showed blood glucose amelioration and improved sperm quality, as well as testicular damage attenuation.

Cyperus esculentus L. es una planta tuberosa que promueve y protege las funciones reproductivas, que generalmente se ven afectadas en los diabéticos. El presente estudio investigó el efecto del extracto de tubérculo de Cyperus esculentus (CETE) sobre la histología testicular y la viabilidad de los espermatozoides de ratas wistar con hiperglicemia inducida por alloxan. Veinticinco ratas Wistar macho adultas que pesaban 150-200 g y se agruparon en cinco (n = 5): el grupo 1, el control, administró agua del grifo (20ml / kg), mientras que los grupos 2-5 se les administró una dosis intraperitoneal única (120 mg / kg p.v.) de alloxan, y agua del grifo por vía oral (20ml/kg), CETE (100 mg/kg), CETE (500 mg/kg) y metformina (500 mg/kg), respectivamente durante 21 días. Los animales fueron sacrificados, su esperma recolectada para su análisis, mientras que los testículos fueron retirados y procesados para histología. Los resultados mostraron un aumento significativo (p<0,05) de la glucosa en sangre y la testosterona, y una disminución significativa (p<0,05) del pH, la motilidad, el recuento, la morfología y la densidad de los espermatozoides, así como interrupciones e hipertrofia de las células espermatogénicas y sertoli del grupo hiperglucémico. Hubo una disminución significativa (p<0,05) de la glucosa en sangre, mientras que los parámetros espermáticos y el peso testicular mejoraron con la histología testicular normal en los grupos de 100 mg / kg de CETE, 500 mg / kg de CETE y tratados con metformina en comparación con el grupo de control e hiperglucémico. El tratamiento con CETE mostró una mejora de la glucosa en sangre y una mejora de la calidad de los espermatozoides, así como atenuación del daño testicular.

Is there a relationship between infertility and fertilin ß protein distribution?

INTRODUCTION: Fertilin ß is a sperm surface protein that can mediate sperm-egg membrane interaction. This study was conducted to determine whether the expression of fertilin ß after intrauterine insemination (IUI) in donors with normal parameters after standard semen analysis is related to low success rate or failure of fertilization. METHODS: We examined the sperm of 30 male donors who have normal as controls, oligozoospermia, and unexplained infertility as the clinically indication for IUI. Fertilin ß has been labeled with the ADAM2 antibody by indirect immunofluorescence (IF) assay. To evaluate the reproducibility of the test, we selected four sperm samples scale of 0 to +++ according to the distribution of fluorescence label. RESULTS: The results were highly correlated with the corrected total cell fluorescence (CTCF) (Rp=0.9972, P<0.05). We suggest that the relationship between infertility and fertilin ß may be due to the distribution of this protein on the sperm surface. Male partners of couples with unexplained infertility showed a low distribution of fertilin ß by a decrease of the fluorescence signal in the IF labeling (scale of +++ by 7.4±10.32%, P<0.0001, ±SD). DISCUSSION: Abnormal fertilin ß function may be a potential mechanism that could lead to fertilization failure.

Cyclic adenosine monophosphate (cAMP) analog and phosphodiesterase inhibitor (IBMX) ameliorate human sperm capacitation and motility.

OBJECTIVES: Human sperm quality is decreasing progressively. One of the foremost reasons for infertility is the failure in sperm capacitation. We examined the influence of a cAMP (cyclic-adenosine mono phosphate analog)+IBMX (3-isobutyl-1-methylxanthine) on the motility and capacitation rate of human sperm over time. MATERIAL AND METHODS: Samples were gotten from 20 asthenozoospermic infertile patients referring to the Academic Center for Education, Culture and Research unit of the infertility research center, Qom, Iran. Samples were processed with a Density Gradient Centrifuging. Spermatozoa were divided into 4 groups: control, experimental 1, 2 and 3 (E1, E2, E3) based on the dose/time schedules (cAMP 5mmol+IBMX 0.2mmol/2, 4, and 6h, respectively). The computer-assisted sperm analysis and chlortetracycline assays were used to measure sperm motility and capacitation. RESULTS: After incubation with a cAMP analog and IBMX, the levels of progressive motile sperms considerably improved in all experimental groups compared to the control group (E1=18.89±7.1, E2=30±9.7, E3=26.3±9.6 vs Control=10.28±6.2, P<0.05) especially in E2 group (P<0.05), indicating a greater effect of db cAMP (5mmol) and IBMX (0.2mmol) for 4h compared to the same doses at 2 and 6h. Also, non-progressive motile sperms significantly decreased in E2 group compared to the other groups (P<0.05). Moreover, both patterns C and B were substantially improved in all experimental groups especially in E2 group (P<0.05). CONCLUSION: Our findings support that the supplementation of sperm with db cAMP+IBMX specially for 4h, could be useful for men with asthenozoospermia to improve the success of assisted reproductive technology.

The effects of wireless devices on male reproductive health: A literature overview / Los efectos de los dispositivos inalámbricos en la salud reproductiva masculina: una descripción general de la literature

Purpose: The aim of this study was to summarize the evidence of radiofrequency electromagnetic radiation (RF-EMR) exposure from wireless devices on total motile sperm count (TMSC) and identify gaps in the literature that could help clarify this link.Materials and methods: A literature search was conducted using PubMed/MEDLINE to find relevant studies examining the effects of EMR on male fertility, with a specific focus on TMSC, published from 2000 to 2019. R was used for data analyses.Results: Motility was identified as the parameter linked to TMSC that was most negatively impacted by EMR exposure. Many gaps were found including geographic and lack of standardization with EMR factors such as exposure time and operating frequency.Conclusion: The EMR emitted by wireless devices may negatively affect TMSC, which is one of the better predictors of achieving pregnancies and impairs male fertility. Our findings highlight the need for clinicians to explore wireless device usage to help guide treatment decisions in men or couples with subfertility concerns. (AU)

Objetivo: El objetivo de este estudio fue resumir la evidencia de la exposición a la radiación electromagnética (EMR) por radiofrecuencia de dispositivos inalámbricos en el recuento total de espermatozoides móviles (TMSC) e identificar brechas en la literatura que podrían ayudar a aclarar este vínculo.Materiales y métodos: Se realizó una búsqueda de literatura en PubMed/MEDLINE para encontrar estudios relevantes que examinaran los efectos de la EMR en la fertilidad masculina, con un enfoque específico en el TMSC, publicados desde 2000 hasta 2019. Se utilizó el programa R para el análisis de datos.Resultados: La motilidad se identificó como el parámetro vinculado al TMSC que se vio más negativamente afectado por la exposición a EMR. Se encontraron muchas lagunas, incluyendo la estandarización geográfica y la falta de estandarización con factores EMR, como el tiempo de exposición y la frecuencia de funcionamiento.Conclusión: La EMR emitida por dispositivos inalámbricos puede afectar negativamente al TMSC, que es uno de los mejores predictores para lograr embarazos y afecta la fertilidad masculina. Nuestros hallazgos ponen de relieve la necesidad de que los médicos exploren el uso de dispositivos inalámbricos para ayudar a guiar las decisiones de tratamiento en hombres o parejas con problemas de subfertilidad. (AU)

Can PCNA and LIM15 gene expression levels predict sperm retrieval success in men with non-obstructive azoospermia?

INTRODUCTION AND OBJECTIVES: It is necessary to be able to predict sperm retrieval before microdissection testicular sperm extraction (mTESE) in azoospermic men. This study established the importance of proliferating cell nuclear antigen (PCNA) and LIM15 gene expression levels in predicting the success of sperm retrieval by mTESE. MATERIALS AND METHODS: One hundred and forty-three men who were diagnosed with non-obstructive azoospermia (NOA) were included in the study. Patients' age, total testosterone and follicle stimulating hormone values, testicular volume and testicular histology were recorded by prospectively. PCNA and LIM15 gene expression levels were determined by real-time PCR in the materials from both ejaculate and testicular specimens. RESULTS: Testis volume and histology were the most important factors in predicting the sperm retrieval rate (SRR). The PCNA and LIM15 gene expression levels measured in testicular tissues and the LIM15 gene expression levels measured in ejaculate significantly correlated with the SRR in mTESE (p=0.038, p=0.022, and p=0.004, respectively). Although the PCNA gene expression level measured in ejaculate was higher in men with successful sperm retrieval, the difference was not statistically significant (p=0.061). According to the multivariate logistic regression analysis, testicular volume and LIM15 gene expression level in ejaculate were independent predictive parameters for sperm retrieval. CONCLUSION: The data showed that LIM15 gene expression level in ejaculate is a useful molecular marker to predict the SRR before mTESE.

The effects of wireless devices on male reproductive health: A literature overview.

PURPOSE: The aim of this study was to summarize the evidence of radiofrequency electromagnetic radiation (RF-EMR) exposure from wireless devices on total motile sperm count (TMSC) and identify gaps in the literature that could help clarify this link. MATERIALS AND METHODS: A literature search was conducted using PubMed/MEDLINE to find relevant studies examining the effects of EMR on male fertility, with a specific focus on TMSC, published from 2000 to 2019. R was used for data analyses. RESULTS: Motility was identified as the parameter linked to TMSC that was most negatively impacted by EMR exposure. Many gaps were found including geographic and lack of standardization with EMR factors such as exposure time and operating frequency. CONCLUSION: The EMR emitted by wireless devices may negatively affect TMSC, which is one of the better predictors of achieving pregnancies and impairs male fertility. Our findings highlight the need for clinicians to explore wireless device usage to help guide treatment decisions in men or couples with subfertility concerns.

Fluoride-induced testicular degeneration and sperm quality deteriorations: Salutary role of Cyperus esculentus tubers (tiger nut) extract in animal model.

OBJECTIVE: Chronic exposure to fluoride causes tissue damage induced by oxidative imbalance, Cyperus esculentus (CE) possess anti-inflammatory and immunostimulatory properties. This study focused on Salutary role of Cyperus esculentus in sodium fluoride (NaF) induced testicular degeneration and sperm quality deteriorations. METHODS: Sexually mature male Sprague-Dawley rats were randomly divided into four groups (n=6). Animals in control group received 2 mls of normal saline per day; CE group received 500mg/kg bw of CE; NaF group received 5mg/kg bw of NaF; NaF+CE group received 500mg/kg bw of CE (for 14 days pre-treatment) and NaF co-treatment till 56 days via gastric gavage. Parameters tested include: testicular histology, sperm parameters, sex hormone, fertility test, malondialdehyde (MDA), superoxide dismutase (SOD), reduced glutathione, glutathione peroxidase (GPX), catalase (CAT), testicular fluoride and testicular cholesterol. RESULTS: Sodium fluoride significantly (p<.05) decrease testicular antioxidant (SOD, CAT, GSH and GPx), sperm quality, hormone profiles (TT, FSH, LH, estrogen levels), testicular cholesterol, morphometric parameters, Johnsen's Score and number of implantations in female rats with corresponding (p<.05) increase in oxidative stress makers and abnormal sperm morphology. Also depleted seminiferous epithelium and degenerate spermatogenic cells. Pretreatment with 500mg/kg bw of CE lowered NaF toxicity by significantly reducing the lipid peroxidation products, fluoride accumulation in the testis, histopathological changes of the testes and spermatozoa abnormalities and reverted observed NaF-induced inhibition in antioxidant parameters and weight of accessory sex organs. CONCLUSIONS: Cyperus esculentus attenuated NaF-induced testicular injuries and protected the seminiferous epithelium, reduced oxidative stress and promoted spermatogenesis.

Crioconservación seminal en peces de agua dulce: aspectos biotecnológicos, celulares y bioquímicos / Seminal Cryopreservation in Freshwater Fish: Biotechnological, Cellular and Biochemical Aspects / Criopreservação Seminal em Peixes de Água Doce: Aspectos Biotecnológicos, Celulares e Bioquímicos

Resumen La crioconservación es una herramienta biotecnológica que en peces está orientada principalmente a la conservación criogénica de semen como estrategia de preservación del recurso genético y a su uso para la producción de alevinos con fines diferentes. Actualmente, los protocolos de crioconservación seminal en peces de agua dulce establecen una amplia variedad de procedimientos cuya efectividad se basa en aspectos ligados a la calidad seminal post-descongelación y la fertilidad, así como su relación con el desarrollo de la progenie. El efecto de la conservación del semen en nitrógeno líquido por periodos amplios de tiempo también toma importancia en ésta biotecnología. Por lo anterior, el objetivo de la presente revisión es describir aspectos biotecnológicos, celulares y bioquímicos asociados al proceso de crioconservación seminal en peces dulceacuícolas, resaltando los avances, las limitaciones y sus perspectivas.

Abstract Cryopreservation is a biotechnological tool that in fish is mainly aimed at cryogenic conservation of semen as a strategy for preserving the genetic resource and its use for the production of fingerlings with different purposes. Currently, seminal cryopreservation protocols in freshwater fish establish a wide variety of procedures whose effectiveness is based on aspects linked to seminal post-thaw quality and fertility, as well as its relationship with the development of the progeny. The effect of preserving semen in liquid nitrogen for extended periods of time also plays an important role in this biotechnology. Therefore, the objective of this review is to describe biotechnological, cellular and biochemical aspects associated with the seminal cryopreservation process in freshwater fish, highlighting the advances, limitations and perspectives.

Resumo A criopreservação é uma ferramenta biotecnológica que em peixes visa principalmente a conservação criogênica do sêmen como estratégia para a preservação do recurso genético e sua utilização para a produção de alevinos para diferentes fins. Atualmente, os protocolos de criopreservação seminal em peixes de água doce estabelecem uma ampla variedade de procedimentos cuja eficácia se baseia em aspectos relacionados à qualidade e fertilidade pós-descongelamento seminal, bem como sua relação com o desenvolvimento da progênie. O efeito da preservação do sêmen no nitrogênio líquido por longos períodos de tempo também desempenha um papel importante nessa biotecnologia. Portanto, o objetivo desta revisão é descrever aspectos biotecnológicos, celulares e bioquímicos associados ao processo de criopreservação seminal em peixes de água doce, destacando os avanços, limitações e perspectivas.

The effects of propofol and midazolam on rat spermatogenic parameters / [Efeitos do propofol e midazolam em parâmetros espermatogênicos em ratos]

There is a need for various anesthetic agents to obtain sperm in the field of human and veterinary medicine. Propofol and midazolam are among the most preferred among these agents. The aim of this study was to determine how sperm paramaters are affected according to the anesthetic agent used. Propofol (2mg/kg) and midazolam (3,5-7,5mg/kg) were administered twice a day (morning-evening) for one week. As a result of this study, there was no statistical difference in sperm density and abnormal sperm rates (respectively P=0,673, P=0,479). Sperm motility rates are similar in the control and propofol groups, while the motility rate in the midazolam group is statistically lower. (Control group %85 - Midazolam group %68.75 - Propofol group %83.75), (P<0.05). As a result of this study, the confidence interval of propofol was higher than the other anesthetic agents used for sperm retrieval.(AU)

São necessários vários agentes anestésicos para obter espermatozoides no campo da medicina humana e veterinária. Propofol e midazolam estão entre os agentes preferidos. O objetivo deste estudo foi determinar como os parâmetros de esperma são afetados de acordo com o agente anestésico utilizado. Propofol (2mg / kg) e midazolam (3,5-7,5mg / kg) foram administrados duas vezes ao dia (manhã e noite) durante uma semana. Neste estudo, não houve diferença estatística na densidade espermática e nas taxas anormais de espermatozoides (respectivamente P = 0,673, P = 0,479). As taxas de motilidade espermática são semelhantes nos grupos controle e propofol, enquanto a taxa de motilidade no grupo midazolam é estatisticamente menor. (Grupo controle % 85 - grupo midazolam % 68,75 - grupo propofol % 83,75), (P <0,05). Neste estudo, o intervalo de confiança do propofol foi maior do que os outros agentes anestésicos utilizados na recuperação espermática.(AU)

The effect of modifiable lifestyle factors on semen quality.

INTRODUCTION AND OBJECTIVES: To examine the association between lifestyle factors (body mass index, smoking, alcohol consumption, coffee intake, physical activity, sauna and cell phone usage, wearing tight-fitting underwear), and conventional semen parameters. MATERIALS AND METHODS: 1311 participants who attended the Andrology Clinic were included in the study. All participants were separated into two groups as men with normozoospermia and dysspermia. All participants answered a questionnaire which contains questions about the modifiable lifestyle factors. The total risk scores were calculated after all the positive lifestyle factors had been counted. RESULTS: Men with normozoospermia and dysspermia consisted of 852 (65.0%) and 459 (35.0%) participants respectively. A negative relationship between the wearing of tight underwear and having normal semen parameters was detected between the two groups (p=0.004). While going to a sauna regularly was negatively related to semen concentration, wearing tight underwear was also related to both lower motility, normal morphology as well as semen concentration (p<0.05). While the total score of all participants was 5.22±1.34 point, there were no statistical differences between the two groups (p=0.332). It was found that having 3 more or fewer points was not related to any type of semen parameters and results of a spermiogram. CONCLUSION: The clinicians should give advice to infertile male patients about changing their risky lifestyle, for infertility, to a healthy lifestyle for fertility. Better designed studies, with larger sample sizes using conventional semen analysis with sperm DNA analysis methods, should be planned to identify the possible effects of lifestyle factors on semen quality.

Is there any relationship between human sperm parameters and protamine deficiency in different groups of infertile men?

OBJECTIVE: Abnormality in Histone-Protamine replacements has been indicated to cause sperm DNA damage and infertility. The aim of the present study was to investigate the relationships between sperm parameters in oligospermia, asthenospermia, and teratospermia with protamine deficiency in infertile men. MATERIAL AND METHOD: In this case-control study, we had three experimental groups including oligospermia (n=100), asthenospermia (n=100), and teratospermia (n=100) as well as normospermia (n=100) as controls. Sperm analyses were performed according to the recommendations of the World Health Organization (WHO, 2010) and sperm chromatin quality was assessed using Chromomycin A3 (CMA3) staining for each sample. RESULTS: The comparison of the data between groups indicated that the percentage of spermatozoa with protamine deficiency was significantly different in patients with oligospermia, asthenospermia, and teratospermia when compared with control ones. However, there was no significant correlation between sperm nuclear protamine deficiency and their parameters of the men with teratospermia using CMA3 test. Regarding the oligospermia and asthenospermia semen samples, the findings showed the negative correlations between the sperm nuclear protamine deficiency and progressive motility as well as immobility (p<0.001). CONCLUSION: The higher proportion of spermatozoa with abnormal chromatin packaging was observed in asthenospermic samples than those from other experimental groups as well as controls. It seems that normal morphology cannot have a valuable predictive value for good chromatin quality of spermatozoa, as much as normal motility characteristics, since samples with high mobility rates often have lower protamine deficiencies. The findings may provide a supportable promoting the future wider clinical application of chromatin/DNA integrity testing along with the semen analysis in male infertility.