Intracerebroventricular injection of kisspeptin in male rats activates hypothalamo-pituitary-gonadal axis, but not hypothalamo-pituitary-adrenal axis.

Kisspeptin is an important hormone involved in the stimulation of the hypothalamo-pituitary gonadal (HPG) axis. The HPG axis can be suppressed in certain conditions such as stress, which gives rise to the activation of the hypothalamo-pituitary-adrenal (HPA) axis. However, the physiological role of kisspeptin in the interaction of HPG and HPA axis is not fully understood yet. This study was conducted to investigate the possible effects of central kisspeptin injection on HPG axis as well as HPA axis activity. Adult male Wistar rats were randomly divided into seven groups as followed: sham (control), kisspeptin (50 pmol), P234 (1 nmol), kisspeptin + p234, kisspeptin + antalarmin (0.1 µg), kisspeptin + astressin 2B (1 µg), and kisspeptin + atosiban (300 ng/rat) (n = 10 each group). At the end of the experiments, the hypothalamus, pituitary, and serum samples of the rats were collected. There was no significant difference in corticotropic-releasing hormone immunoreactivity in the paraventricular nucleus of the hypothalamus, serum adrenocorticotropic hormone, and corticosterone levels among all groups. Moreover, no significant difference was detected in pituitary oxytocin level. Serum follicle-stimulating hormone and luteinizing hormone levels of the kisspeptin, kisspeptin + antalarmin, and kisspeptin + astressin 2B groups were significantly higher than the control group. Serum testosterone levels were significantly higher in the kisspeptin kisspeptin + antalarmin, kisspeptin + astressin 2B, and kisspeptin + atosiban groups compared to the control group. Our findings suggest that central kisspeptin injection causes activation in the HPG axis, but not the HPA axis in male rats.

The Impact of the Number of Sutures on Regeneration in Nerve Repair.

PURPOSE: The ideal number of sutures for epineural nerve repair is still unclear. Increased number of sutures increases secondary damage and inflammation to the nerve tissue, which negatively affects nerve regeneration. When the number of sutures decreases, the strength of the nerve repair site decreases and nerve endings are fringed, which also negatively affects nerve regeneration. Therefore, each additional suture is not only beneficial but also detrimental. The aim of this study was to find out the ideal number of sutures for nerve repair. METHODS: Seventy rats were randomly divided into 5 groups. One of the groups was used as a control group, and right sciatic nerves of the rats in other 4 groups were repaired by using 2, 3, 4, or 6 epineural sutures, respectively, after nerve transection. Biomechanical assessment was performed on the nerves collected from these rats at 5 days of follow-up. Functional and histological analyses were evaluated after 12 weeks of follow-up. RESULTS: It was found that an increase in the number of sutures enhances resistance to tensile force in general. However, there was no significant biomechanical difference between the 6-sutured group in which the most sutures were used and the 4-sutured group. In functional examinations, overall successful results were obtained in the group with 4 sutures. In histological examinations, there was no statistical difference between the control group, 2-sutured groups, and 4-sutured groups in terms of connective tissue index. However, it was observed that the group with 6 sutures had a higher connective tissue index than the control group and groups with 2 and 4 sutures. In terms of regeneration index, it was found that repair with 4 sutures was superior to repair with 2 and 6 sutures. No difference was found between any of the suture groups according to the diameter change index. CONCLUSIONS: These results indicate that repair with 4 sutures is the best method of epineural repair that provides both strength and regeneration. These findings will contribute to both the repair of clinically similar nerves and the standardization of rat nerve studies.

Effects of Lipokit® centrifugation on morphology and resident cells of adipose tissue / Efectos de la centrifugación de Lipokit® sobre la morfología y las células residentes del tejido adiposo

The aim of adipose tissue engineering is creating autologus vascularized fat tissue to be used for practical soft tissue reconstruction in human clinic. Unfortunately, in practice, long-term results of fat transplantation are often untrustworthy and unreliable, to overcome this problem different many lipoinjection techniques developed in the last 20 years. Centrifuge is a fundamental stepin the preparation of adipose tissue. We focused on some cell markers especially MSCs markers and histological structural properties after with lipokit centrifugation and without lipokit centrifugation of adipose tissue obtained by liposuction by this new technique. Adipose tissue was taken by liposuction and separates to two portions. One of them is centrifugated by Lipokit machine (C+) has a micro filter and the other is not (C-). After centrifugation smear slides and paraffin sections were prepared from these tissues. These slides were stained with H&E and Toluidine Blue. Paraffin sections were immunohistochemically stained with CD34, von Willebrand Factor, CD73, CD90 and CD105. Smear preparations showed a continuous three dimensional plasma membrane appearance of adipocytes. C+ and C- showed expression of CD34, von Willebrand Factor, CD73, CD90, CD105. C+ seems to have more free cells expressing than C-. While passing the filter of Lipokit, large adipocytes and connective tissue parts disintegrate and thus increases the surface area of lipoaspirate. Lipokit® machine release the group cells which are necessary for angiogenesis and they become more freely to construct angiogenesis.

El objetivo de la ingeniería del tejido adiposo es la creación de tejido graso vascularizado autólogo para ser utilizado en clínica humana para la reconstrucción de tejido blando. Desafortunadamente en la práctica, los resultados a largo plazo del trasplante de grasa son poco fiables y no seguros; para superar este problema, se han desarrollado en diferentes países, en los últimos 20 años, variadas técnicas de lipoinyección. La centrifugación es un paso fundamental en la preparación del tejido adiposo. Nos hemos centrado en algunos marcadores, especialmente, de células precursoras mesenquimales y propiedades histológicas estructurales después de la centrifugación mediante Lipokit y sin la centrifugación por Lipokit del tejido adiposo obtenido mediante liposucción. El tejido adiposo fue tomado por liposucción y se separó en dos porciones. Una se centrifugó mediante el sistema Lipokit (C +), con un microfiltro y la otro no (C-). Después de centrifugación, muestras del frotis y secciones de parafina se prepararon a partir de estos tejidos. Los frotis se tiñeron con H&E y azul de toluidina. Las secciones de parafina se tiñeron inmunohistoquímicamente con CD34, factor de von Willebrand, CD73, CD90 y CD105. Las preparaciones de los frotis mostraron una apariencia tridimensional continua de la membrana plasmática de los adipocitos. Tanto en C+ y C- se observó la expresión de CD34, factor de von Willebrand, CD73, CD90 y CD105. En C+ parecen expresarse más células libres que en C-. Cuando se utilizó el filtro de Lipokit, los adipocitos grandes y partes del tejido conectivo se desintegraron, por lo tanto aumentó el área de superficie de lipoaspirado. El sistema Lipokit® libera los grupos celulares que son necesarios para la angiogénesis y se hacen más libres para promoverla.

Assessment of Spermatozoa Morphology under Light Microscopy with Different Histologic Stains and Comparison of Morphometric Measurements / Evaluación de la Morfología de los Espermatozoides bajo Microscopía Óptica con Diferentes Tinciones Histológicas y Comparación de las Mediciones Morfométricas

The aim was to examine the morphology of spermatozoa with different staining methods and aimed to find the better staining methods for morphology of spermatozoa in our study. Randomized 67 patients taken for the study who were admitted to Assisted Reproductive Techniques Unit. In the first part of the study, smears were stained with Hematoxylin Eosin (HE), Toluidin Blue (TB), Giemsa, Wright, ferrous Weigert haematoxylin stain, Orange G, eosin-aniline blue dye, Shorr Method, Papanicolau, Berg Method, Light Green stain, Acridine Orange (AO) and Janus Green dyes. In the second part of the study, smear preparations of 10 patients with normozoospermic were stained with HE, Toluidin Blue (TB), Shorr Method and Papanicolau. Four measurements were made including the middle piece, head length- head width and tail length for 200 spermatozoa with normal morphology. Comparisons were made between the stains that which showed a better morphology. Condensation assessment was not possible in smears stained with Shorr, Berg, Method, AO. Better assessment of condensation could be made in other stains. In the second part the smallestvalues belonged to of TB stain according to measurements of head of the spermatozoa. There was a significant difference at the head length with TB stain. Although measurements of Shorr and Papanicolau areclose to each otherand the largestvalues belonged to Papanicolau dye. It was concluded that measurement values in human sperm morphology could alter with the used staining method.

El objetivo fue examinar la morfología de los espermatozoides con diferentes métodos de tinción y encontrar los mejores métodos para su estudio. Fueron seleccionados para el estudio, de manera aleatoria 67 pacientes, quienes ingresaron a la Unidad de técnicas de reproducción asistida. En la primera parte del estudio, se realizaron y tiñeron frotis con hematoxilina eosina (HE), Azul de Toluidina (AT), Giemsa, tinción de Wright, Hematoxilina Férrica de Weigert, Anaranjado G, tinción eosina-anilina, método de Shorr, Papanicolau, método Berg, tinción verde brillante, anaranjado de acridina (AO) y tinción verde Janus. En la segunda parte del estudio, se realizaron frotis de 10 pacientes con normozoospérmicos y se tiñeron con HE, AT, Método Shorr y Papanicolau. Se realizaron cuatro mediciones: ancho de la cabeza, longitud de la cabeza, parte media y cola, sobre 200 espermatozoides con morfología normal. Se compararon las tinciones que mostraban mejor la morfología. La evaluación de la compactación no fue posible en los frotis teñidos con los métodos de Shorr, Berg y AO. Una mejor evaluación de la copactación podría hacerse en otras tinciones. En la segunda parte los valores menores correspondieron a la tinción de AT en relación a la medición de la cabeza de los espermatozoides. Hubo una diferencia significativa en la longitud de la cabeza con tinción de AT. Las mediciones en los frotis con técnicas de Shorr y Papanicolau fueron similares, con valores más altos bajo tinción de Papanicolau. Se concluyó que los valores de la medición morfológica en espermatozoides humanos podrían ser alterados según el método de tinción utilizado.