Comparison of male reproductive parameters in mice with type 1 and type 2 diabetes.

OBJECTIVE: The differences between type 1 and type 2 diabetes mellitus (T1DM and T2DM) in terms of their adverse effects on male reproductive parameters have never been elucidated. This study aimed to distinguish between the effects of the DM types in mice treated with multiple low doses of streptozotocin (STZ) to mimic human T1DM and coadministered a high-fat diet (HFD) to mimic human T2DM. METHODS: The T1DM mice were intraperitoneally injected with STZ (40 mg/kg body weight) for 5 days. The T2DM mice received an HFD for 14 days prior to STZ injection (85 mg/kg body weight), followed by continuous feeding of an HFD. Male reproductive parameters were evaluated. RESULTS: The reproductive organs of the DM mice weighed significantly less than those of controls, and the seminal vesicles plus prostates of the T1DM mice weighed less than those of the T2DM mice. Increased sperm abnormalities and incomplete DNA packaging were observed in the DM groups. Sperm concentration and the proportion of normal sperm were significantly lower in the T1DM group. The seminiferous histopathology of DM mice was classified into seven types. The penises of the DM mice were smaller than those of the controls; however, tunica albuginea thickness and the amount of penile collagen fibers were increased in these mice. Round germ cells were abundant in the epididymal lumens of the mice with DM. CONCLUSION: T1DM adversely affected reproductive parameters to a greater extent than T2DM.

Expression of testicular phosphorylated proteins in types 1 and 2 diabetes mellitus in mice: An experimental study.

BACKGROUND: Types 1 and 2 diabetes mellitus (DM) are known to be the cause of sub/infertility. However, the comparisons of potential markers in spermatogenesis and steroidogenesis in DM males have never been elucidated. OBJECTIVE: This study aimed to examine the expressions of tyrosine-phosphorylated and steroidogenic acute regulatory (StAR) proteins in testis of DM mice. MATERIALS AND METHODS: Fifty-six male C57BL/6 mice were divided into four groups (n░=░14/ each): control of MLD-STZ (multiple low doses of streptozotocin), MLD-STZ, control of HFD-STZ (high-fat diet with STZ), and HFD-STZ. MLD-STZ mice (type 1 DM) were intraperitoneally (i.p.) injected with STZ at 40░mg/kg BW for five days. HFD-STZ mice (type 2 DM) received an HFD for 14 days and i.p.-induced by STZ at 85░mg/kg BW and fed with HFD. At the end of the experiment (days 36 and 72), the expressions of phosphorylated proteins and StAR were examined. RESULTS: Tyrosine phosphorylated proteins were localized in late spermatids, luminal fluid, and Leydig cells. The intensities of phosphorylated 110, 85, 72, 60, and 55░kDas were lower in the 36 day-DM mice. Although such intensities were present in both groups, only 85░kDa in the MLD-STZ mice was higher in HFD mice at 72 days. StAR expressions in both groups were decreased than that of the controls. CONCLUSION: Decreased expressions of StAR and tyrosine-phosphorylated proteins may be directly involved in low testosterone levels and impaired spermatogenesis. These findings support the notion that both DM types play a role in male infertility.

Testicular histopathology and phosphorylated protein changes in mice with diabetes induced by multiple-low doses of streptozotocin: An experimental study.

BACKGROUND: The streptozotocin (STZ)-induced diabetic model is widely used to evaluate the adverse effects of diabetes mellitus (DM) on spermatogenesis and testicular steroidogenesis. However, the actual mechanism of sub/infertility in DM males needs to be elucidated. OBJECTIVE: To conduct a detailed examination of the testicular histopathology, sperm acrosome reaction (AR) status, and tyrosine-phosphorylated protein expression in the testis of male mice induced with STZ. MATERIALS AND METHODS: Ten ICR mice were divided into two groups (n=5/each): control and diabetes induced by multiple low doses of streptozotocin (MLD-STZ). The control mice were intraperitoneally injected with citrate buffer, whereas MLD-STZ mice were injected with STZ at 40 mg/kg body weight for five consecutive days. At the end of the experiment (day 40), reproductive parameters, AR status, and the histopathology of the testis and epididymis were evaluated. The expression of testicular tyrosine phosphorylated proteins was examined. RESULTS: Blood glucose levels, AR percentages, and sperm abnormality of STZ group were significantly higher (p=0.003, 0.001, 0.000), while sperm concentration was significantly lower (p=0.001) compared to control. Histopathology of the seminiferous tubule was classified into 7 types. Additionally, abundant round cells were found in the epididymal lumen of the MLD-STZ mice. Moreover, the intensities of testicular phosphorylated proteins (170, 70, 36, 30, and 25 kDas) were markedly higher and a 120 kDa protein band was noticeably lower in the MLD-STZ mice. CONCLUSION: MLD-STZ-induced DM causes many testicular histopathologies, precocious sperm AR, and increased expression of testicular phosphorylated proteins. These findings may clarify some mechanisms of sub/infertility in DM males.

Antioxidant capacity of Momordica charantia extract and its protective effect on testicular damage in valproic acid-induced rats / Capacidad antioxidante del extracto de Momordica charantia y su efecto protector sobre el daño testicular en ratas inducidas con ácido valproico

Although Momordica charantia (MC) has preventive effects on tissue injuries, antioxidant capacity and protective effect of MC pulp and peel (MCP) on valproic acid (VPA)-testicular damage have never been reported. Fresh MCPs were aqueous extracted and determined for antioxidant capacity and momordicine I level by HPLC. Male rats were divided into 5 groups (control, VPA (500 mg/kgBW), MCP20/40/ or 80 mg/kgBW+VPA). In 30 experimental days, animals were pretreated with different doses of MCPs for 20 days before VPA injection for 10 consecutive days. Sperm concentration, testosterone hormone, and testicular histology of all groups were investigated. Expressions of testicular tyrosine phosphorylated and steroidogenic acute regulatory (StAR) proteins were examined by Western blot. Results showed that MCP contains TPC (39.24±0.65 ug/mg garlic acid), antioxidant capacities (FRAP=33.08±0.21 ug/ mg ascorbic acid equivalent, IC50 of DPPH=389.8±3.20 ug/ml), and momordicine I (404.9 mg/g MCP). Sperm concentration in MCP80+VPA group was increased as compared to VPA group. Testosterone level in MCP treated groups was significantly increased. MCP protected testicular damage and could prevent the decrease of StAR and a 50-kDa phosphorylated protein expression in VPAtreated testis. In conclusion, MCP has antioxidant activities and can prevent male reproductive toxicity in VPA-induced rats.

A pesar que la Momordica charantia (MC) tiene efectos preventivos sobre las lesiones en los tejidos, capacidad antioxidante y un efecto protector de la pulpa y la cáscara de MC (CMC) sobre el ácido valproico (AVP), aún no se ha informado efectos sobre el daño testicular. Las CMC frescas fueron extraídas de forma acuosa y se determinó la capacidad antioxidante y el nivel de Momordicina I por HPLC. Las ratas machos se dividieron en 5 grupos: control, AVP (500 mg/kg de peso corporal), CMC20 / 40 / u 80 mg/kg de peso corporal + AVP . En 30 días experimentales, los animales fueron pretratados con diferentes dosis de CMC durante 20 días antes de la inyección de AVP durante 10 días consecutivos. Se investigó la concentración de espermatozoides, la hormona testosterona y la histología testicular de todos los grupos. Las expresiones de proteínas reguladoras agudas (StAR) fosforiladas con tirosina y esteroidogénicas testiculares se examinaron mediante inmunotransferencia de tipo Western. Los resultados mostraron que CMC contiene TPC (39.24 ± 0.65 ug / mg de ácido de ajo), capacidades antioxidantes (FRAP = 33.08 ± 0.21 ug / mg de ácido ascórbico equivalente, IC50 de DPPH = 389.8 ± 3.20 ug / ml) y momordicina I (404.9 mg) / g CMC). La concentración de esperma en el grupo MCP80 + AVP aumentó en comparación con el grupo AVP. El nivel de testosterona en los grupos tratados con CMC aumentó significativamente. La CMC protegió el daño testicular y pudo prevenir la disminución de StAR y una expresión de proteína fosforilada de 50 kDa en los testículos tratados con AVP. En conclusión, la CMC tiene efectos antioxidantes y puede prevenir la toxicidad reproductiva en ratas machos inducidas por VPA.

Antioxidant and hypoglycemic effects of Momordica cochinchinensis Spreng: (Gac) aril extract on reproductive damages in streptozotocin (STZ)-induced hyperglycemia mice / Efectos antioxidantes e hipoglucémicos del extracto de aril de Momordica cochinchinensis Spreng: (Gac) sobre los daños reproductivos en ratones con hiperglucemia inducida por streptozotocin (STZ)

The aim of present study was to investigate the effect of Momordica cochinchinensis (Gag) aril (GA) aqueous extract on male reproductive system of streptozotocin (STZ)-induced hyperglycemia (HG) mice. GA were extracted with distilled water (DW) and analyzed for in vitro antioxidant capacities. ICR male mice were divided into 7 groups: 1) control, 2) DW, 3) GA 1000 mg/kg BW, 4) HG, 5) HG + glibenclamide, 6 and 7) HG + GA 500 and 1000 mg/kg BW respectively (7 mice/ group). In HG groups, mice were induced by STZ at single dose (150 mg/kg BW). They were treated for consecutive 35 days. All groups were compared for blood glucose levels, weights and histopathologies of reproductive organs, sperm concentration including testicular tyrosine phosphorylation protein patterns by Immuno-Western blotting. The results showed that GA processed antioxidant activities and could significantly decrease blood glucose levels and increase sperm concentration in HG mice. Moreover, GA could change the density of a testicular 70 kDa protein in HG-GA groups. In conclusion, GA extract could improve hyperglycemia and male reproductive damages in STZ-induced HG mice.

El objetivo de este estudio fue investigar el efecto del extracto acuoso de Momordica cochinchinensis (Gag) aril (GA) en el sistema reproductor masculino de ratones hiperglucémicos inducidos por estreptozotocina (STZ). GA fue extraída con agua destilada (DW) y se analizaron las capacidades antioxidantes in vitro. Ratones ICR machos fueron divididos en 7 grupos: 1) control, 2) DW, 3) GA 1000 mg / kg PC, 4) HG, 5) HG + glibenclamida, 6 y 7) HG + GA 500 y 1000 mg / kg PC, respectivamente (7 ratones / grupo). En los grupos HG, los ratones fueron inducidos con STZ en dosis única (150 mg / kg BW). Fueron tratados durante 35 días consecutivos. En todos los grupos se compararon los niveles de glucosa en sangre, los pesos y las histopatologías de los órganos reproductores, la concentración de espermatozoides, incluídos los patrones testiculares de proteínas tirosina fosforilada por Inmuno-Western blot. Los resultados mostraron que GA procesaba actividades antioxidantes y podían disminuir significativamente los niveles de glucosa en sangre y aumentar la concentración de espermatozoides en ratones HG. Además, GA podría cambiar la densidad de una proteína testicular de 70 kDa en grupos HG-GA. En conclusión, el extracto de GA podría mejorar la hiperglucemia y los daños reproductivos masculinos inducidos por STZ en ratones HG.

Morphometry of Isan-Thai clavicles as a guide for sex determination / Morfometría de las clavículas en una población Isan-Thai como guía para la determinación del sexo

The dried clavicles have been well documented to determine individual sex and age in many races. Such morphometric investigations in Isan (Northeastern) Thais have never been reported. Therefore, this study aimed to estimate the sex from dried clavicles of Isan-Thais using morphometric analysis. The identified 454 dried clavicles (254 males and 200 females), averaged age (60.69±14.36 years) were measured and analyzed for identification point (IP) and the demarking point (DP) values. The results showed that the identified-bone percentages by IP of the maximum clavicular lengths in males and females were 90.55 % (>139.9 mm) and 89 % (<140.6 mm). In addition, such percentages of the mid shaft circumference measured in male clavicles was 83.46 % (>37.7 mm) and in females was 75 % (<37.0 mm). However, percentages of the DP in all parameters were less than 50%. The highest DP values of maximum length of male clavicles was 43.31% (>152.5). In conclusion, the maximum length and mid shaft circumference investigated from this study can be used as basic data of Isan ­ Thais for applying as a guide in forensic sciences for sex determination from dried clavicle remains.

Las clavículas secas han sido bien documentadas en muchas razas para determinar el sexo y la edad individuales. Tales investigaciones morfométricas en Isan (Noreste de Tailandia) nunca han sido reportadas. Por lo tanto, este estudio tuvo como objetivo estimar el sexo de clavículas secas de Isais-Thais utilizando un análisis morfométrico. Se midieron 454 clavículas secas (254 varones y 200 hembras), con una edad media de 60,69 ± 14,36 años, y fueron analizadas para los puntos de identificación (IP) y de demarcación (DP). Los resultados mostraron que los porcentajes de hueso identificados por IP en relación a la longitud clavicular máxima en hombres y mujeres fueron 90,55 % (> 139,9 mm) y 89 % (<140,6 mm), respectivamente. Además, tales porcentajes de circunferencia media del eje, medido en las clavículas de los hombres, fue 83,46 % (> 37,7 mm) y en las de mujeres fue 75 % (<37,0 mm). Sin embargo, los porcentajes de la DP en todos los parámetros fueron menores al 50 %. Los valores más altos de DP de longitud máxima de las clavículas de los hombres fue 43,31 % (> 152,5). En conclusión, la longitud máxima y la circunferencia media del eje investigados a partir de este estudio se pueden utilizar como datos básicos en Isan - Thais para aplicar como guía en las ciencias forenses para la determinación del sexo a partir de restos de clavícula secas.

Incidence of Clavicular Rhomboid Fossa in Northeastern Thais: An Anthropological Study.

The rhomboid fossa of clavicle is used to determine the age and sex in anthropology and forensic sciences. The variant types of rhomboid fossa on inferior surface have been reported in many races except in Thais. This study therefore was aimed at classifying the types of the rhomboid fossa in Northeastern Thais. The identified 476 Northeastern Thais dried clavicles (270 males and 206 females) were observed and recorded for the types of rhomboid fossa. The results showed that Thai-rhomboid fossa could be classified into 4 types: Type 1: smooth; Type 2: flat; Type 3: elevated; and Type 4: depressed, respectively. The incidences of rhomboid fossa were as follows: Type 1: 0.21%; Type 2: 19.75%; Type 3: 76.26%; and Type 4: 3.78%, respectively. Additionally, it was found that the percentage of Type 4 (11.84%) was much greater than that of female (1.94%) compared to other types. This incidence of rhomboid fossa types especially Type 4 may be a basic knowledge to be used in sex identification. The high incidence of rhomboid fossa in both sexes of Northeastern Thai clavicles was Type 3 (elevated type).

Chronic restraint stress induces sperm acrosome reaction and changes in testicular tyrosine phosphorylated proteins in rats.

BACKGROUND: Stress is a cause of male infertility. Although sex hormones and sperm quality have been shown to be low in stress, sperm physiology and testicular functional proteins, such as phosphotyrosine proteins, have not been documented. OBJECTIVE: To investigate the acrosome status and alterations of testicular proteins involved in spermatogenesis and testosterone synthesis in chronic stress in rats. MATERIALS AND METHODS: In this experimental study, male rats were divided into 2 groups (control and chronic stress (CS), n=7). CS rats were immobilized (4 hr/day) for 42 consecutive days. The blood glucose level (BGL), corticosterone, testosterone, acrosome status, and histopathology were examined. The expressions of testicular steroidogenic acute regulatory (StAR), cytochrome P450 side chain cleavage (CYP11A1), and phosphorylated proteins were analyzed. RESULTS: Results showed that BGL (71.25±2.22 vs. 95.60±3.36 mg/dl), corticosterone level (24.33±4.23 vs. 36.9±2.01 ng/ml), acrosome reacted sperm (3.25±1.55 vs. 17.71±5.03%), and sperm head abnormality (3.29±0.71 vs. 6.21±1.18%) were significantly higher in CS group in comparison with control. In contrast, seminal vesicle (0.41±0.05 vs. 0.24±0.07 g/100g), testosterone level (3.37±0.79 vs. 0.61±0.29 ng/ml), and sperm concentration (115.33±7.70 vs. 79.13±3.65×10(6) cells/ml) of CS were significantly lower (p<0.05) than controls. Some atrophic seminiferous tubules and low sperm mass were apparent in CS rats. The expression of CYP11A1 except StAR protein was markedly decreased in CS rats. In contrast, a 55 kDa phosphorylated protein was higher in CS testes. CONCLUSION: CS decreased the expression of CYP11A, resulting in decreased testosterone, and increased acrosome-reacted sperm, assumed to be the result of an increase of 55 kDa phosphorylated protein.

Changes of testicular phosphorylated proteins in response to restraint stress in male rats.

OBJECTIVE: To investigate male reproductive parameters via changes of potential testicular protein markers in restraint-stress rats. METHODS: Male Sprague-Dawley rats were divided into two groups (non-immobilized control and restraint-immobilized/stress groups, n=8 each group). The stress animals were immobilized (12 h/d) by a restraint cage for 7 consecutive days. All reproductive parameters, morphology and histology were observed and compared between groups. In addition, the expression of steroidogenic acute regulatory (StAR) and phosphotyrosine proteins (previously localized in Sertoli and late spermatid cells) in testicular lysate was assayed by immuno-Western blotting. RESULTS: Testosterone level, sperm concentration and sperm head normality of stress rats were significantly decreased while the corticosterone level was increased as compared with the control (P<0.05). Histologically, stress rats showed low sperm mass in epididymal lumen and some atrophy of seminiferous tubules. Although the expression of testicular StAR protein was not significantly different between groups, changed patterns of the 131, 95, and 75 kDa testicular phosphorylated proteins were observed in the stress group compared with the control group. The intensity of a testicular 95-kDa phosphorylated protein was significantly decreased in stress rats. CONCLUSIONS: This study has demonstrated the alteration of testicular phosphorylated protein patterns, associated with adverse male reproductive parameters in stress rats. It could be an explanation of some infertility in stress males.