Effects of 3-Heptyl-3,4,6-trimethoxy-3H-isobenzofuran-1-one alone or/in association with cyclophosphamide on testicular function.

Chemotherapy for cancer treatment may result in a temporary or long-term gonadal damage resulting in subfertility or infertility. Cyclophosphamide (CY) is a cytotoxic alkylating agent that has been widely used in the treatment of cancer. Recent studies have shown that synthetic resorcinol lipid AMS35AA (3-Heptyl-3,4,6-trimethoxy-3H-isobenzofuran-1-one) may be an important adjuvant chemotherapy that potentiates mutagenic damage and increases apoptosis caused by CY. The present study investigates the action of AMS35AA alone or/in association with CY on testicular function. Animals were divided into four groups: (a) control group: received only water; (b) CY group: received 150 µg/g of CY b.w., i.p.; (c) AMS35AA group: received 10 µg/g of AMS35AA b.w., i.p; and (d) associated group: received 10 µg/g of AMS35AA + 150 µg/g of CY b.w., i.p. Four weeks after the treatment, the results showed that testes weight of CY and associated groups decreased. However, the number of Sertoli cell and Leydig cell per testis was similar in control and treated groups. Our findings provide strong evidence that the AMS35AA alone or in CY association is not toxic to spermatogenesis. The absence of toxicity of AMS35AA supports the view that the resorcinolic lipid could be used associated with CY chemotherapy without causing adverse effects to testes function.

Evaluation of the testis function of mice exposed in utero and during lactation to Pfaffia glomerata (Brazilian ginseng).

Pfaffia glomerata (Spreng.) Pedersen, popularly known as "Brazilian ginseng," is used as medicinal plant in Brazil to treat inflammatory diseases in general. Previous studies showed that its extract increases the nitric oxide (NO) levels. Knowing that NO downregulates steroidogenesis and that alterations in the action/production of androgens during perinatal life could alter testis development, the present studies sought to investigate the reproductive toxicity of Pfaffia glomerata on male mice exposed to hydroalcoholic extract in utero and during lactation. The present study shows that P. glomerata extract does not alter body weight, tubular diameter and testis function in male mice. Although a reduction in the testis weight was observed in the animals that received the highest dose directly in early post-natal life, our findings show clearly that P. glomerata may not act as an endocrine disruptor, and it is not an "antiandrogenic" compound that could lead to testicular dysgenesis syndrome.

Comparison of semen samples collected from wild and captive jaguars (Panthera onca) by urethral catheterization after pharmacological induction.

This study was designed to evaluate the efficiency of medetomidine anesthesia for semen collection through urethral catheter in wild and captive jaguars. Six captive and five wild jaguars were chemically restrained with a combination of medetomidine (0.08-0.1 mg/kg) and ketamine (5 mg/kg). After medetomidine administration the urethra was catheterized using a urinary tomcat catheter (1 mm diameter × 130 mm length) to collect semen from all animals. By using this technique, we could collect an average of 347.2 µl of semen containing 2,635.2 sperm/ml. Forward progressive motility, sperm progressive motility, and sperm morphology analysis demonstrated that the methodology did not affect sperm quality. Thus, urethral catheterization after medetomidine administration is a practical and efficient method to collect high-quality semen from wild and captive jaguars; this will enable the development of reproductive assisted technologies for jaguars.

Intake of Polydextrose Alters Hematology and the Profile of Short Chain Fatty Acids in Partially Gastrectomized Rats.

Polydextrose (PDX) ingestion may increase the intestinal absorption of iron. This study evaluated the effects of 7.5% polydextrose supplementation on markers of iron uptake, transport and storage in partially gastrectomized rats. Half of a batch of 40 male Wistar rats (250 g) underwent Billroth II partial gastrectomy with anterior truncal vagotomy (GXT), while the other half underwent sham gastrectomy (SHAM). At 7 postoperative days, the animals were subdivided into four groups (n = 10): Sham Control and GXT Control (no polydextrose); Sham PDX and GXT PDX (with 7.5% PDX). The animals were euthanized after 60 day of PDX treatment. Organ weight, cecal pH, the characterization and quantification of short-chain fatty acids (SCFA), hematological parameters, hepatic iron content and the expression of ferroportin (FPT) in the jejunum, cecum, colon and liver were evaluated. PDX caused changes in the cecum of the supplemented animals, where there was a decrease in pH, increase in cecal wall and marked production of SCFA, especially acetic and propionic acids (p < 0.05). Hepatic iron levels were lower in GXT animals. PDX increased hemoglobin (HGB) values by 29.2% and hematocrit (HCT) by 55.8% in the GXT PDX group compared to the GXT Control group. The GXT PDX group had lower hepatic FPT expression (p < 0.05). PDX led to increased SCFA concentration in the supplemented animals. Considering that SCFAs play a central role in the increasing nutrients uptake, this mechanism may be involved in altering the hematology profile observed in these animals but not enough to reverse iron deficiency anemia in post-gastrectomy rats.

Actions of sesquiterpene lactones isolated from Moquiniastrum polymorphum subsp. floccosum in MCF7 cell line and their potentiating action on doxorubicin.

BACKGROUND: In order to obtain better clinical results in anticancer therapies, polychemotherapy or combination therapies are used. For this, the combinations are required to increase the efficacy and reduce the adverse reactions of the associated chemotherapies. The aim of this study was to evaluate the cytotoxic, apoptotic and (anti)proliferative potential of two sesquiterpene lactones isolated from Moquiniastrum polymorphum, 11,13-diidrozaluzanin C (1) and gochnatiolide C (2), and their associations with chemotherapeutic agents irinotecan, tamoxifen, cisplatin, 5-fluouracyl and doxorubicin in the tumoral lineage of MCF-7 breast adenocarcinoma. METHODS: The analyses were performed by MTT cytotoxicity assays, drug combination index (CI), apoptosis morphological assay and cell proliferation assay. Treatments were evaluated with short exposure times (4 h), followed or not by recovery in drug-free medium for 24 h. For the cell viability assay the statistical analysis was performed using software INSTAT, and the ANOVA/Tukey test was applied. Combination Indices (CI) was made using CompuSyn software and demonstrated through isoboles. The assays that evaluated cell death and proliferation used statistical analysis SAS 9.4 (Statistical Analysis System), and the procedure adopted was PROC NPAR1WAY. The Wilcoxon test at 5% level was applied for comparing statistical differences. RESULTS: The results demonstrated that the compounds decrease cell viability and increase their action when associated with irinotecan, tamoxifen and doxorubicin (CI < 1 and CI = 1). In periods of 4 h-exposure, the compounds cause cell death by apoptosis and after 24 h, they increase the mean number of cells in programmed cell death, especially when treated with 2. In addition, the association with doxorubicin increases the apoptotic potential induced by tested compounds. Both isolates had effect on the reduction of the number of mitoses, especially when 2 at its highest concentration is associated with doxorubicin. CONCLUSIONS: Finally, these compounds are presented as potential agents in chemotherapy combined with doxorubicin, since they trigger the mechanism of apoptosis, which, through the mechanism of action of sesquiterpene lactones, leads to a reduction in toxicity. In addition, the tested compounds have the ability to exert a synergistic action with doxorubicin, possibly by down-regulating the drug resistance mechanisms.

Testicular Histomorphometric Evaluation of Zebu Bull Breeds

The objective of this study was to evaluate the quantitative histology and testicular biometrics in zebu bulls of different breeds. Testicular fragments of Nelore (n=10), Polled Nelore (n=6), Gir (n=5), Guzerat (n=5) and Tabapuã bulls (n=5) were used. The fragments were perfusion-fixed in Karnovsky solution, embedded in glycol methacrylate and stained with toluidine blue-1% sodium borate. The Nelore animals had a higher tubular volumetric proportion (85.2%) and greater height of the seminiferous epithelium (73.2 µm) than the Gir, Guzerat and Tabapuã breeds. The Nelore animals also had a higher volumetric proportion of Leydig cells (5.2%) than the Guzerat and Tabapuã breeds. There was no significant difference for any of these parameters between the Nelore and Polled Nelore breeds. The gonadosomatic index, seminiferous tubule diameter, cross-sectional area of the seminiferous tubule and tubule length (total length and length per gram of testicular parenchyma) did not vary among the breeds studied. The morphometric parameters evaluated suggested that the genetic selection applied to the Nelore and Polled Nelore breeds improved the efficiency of spermatogenesis in these breeders.

Comparação entre duas concentrações de glicerol para a criopreservação de sêmen de suçuarana (Puma concolor) / Comparison between two glycerol concentrations to cryopreservation of semen of mountain lions (Puma concolor)

O desenvolvimento de biotécnicas de reprodução é uma importante ferramenta para a conservação de animais silvestres ameaçados de extinção. Procedimentos de reprodução assistida em suçuarana, no entanto, são escassos na literatura, em especial aqueles relacionados à criopreservação de sêmen. Neste sentido, o presente trabalho objetivou avaliar a congelabilidade do sêmen de suçuaranas adultas mantidas em cativeiro, por meio da comparação entre duas concentrações de glicerol no meio de congelamento. Foram usados cinco machos adultos de suçuarana, mantidos no Centro de Reabilitação de Animais Silvestres do Mato Grosso do Sul (CRAS/MS). As amostras foram coletadas por eletroejaculação e avaliadas quanto ao seu aspecto físico, volume, vigor, motilidade, concentração e índice espermático. De cada ejaculado duas alíquotas foram diluídas em meio Tris-citrato-gema de ovo, em concentrações finais de 5 e 7,5% de glicerol, resfriadas a uma taxa de -0,55ºC/min e congeladas a uma taxa de -5,8ºC/min. Depois de descongeladas, as amostras foram reavaliadas e submetidas aos testes de termorresistência e hiposmótico. O protocolo de criopreservação e descongelamento de sêmen proposto se mostrou eficiente em ambas as concentrações de glicerol testadas, não havendo diferença (p>0,05) entre estas.

The development of biotechnologies of reproduction is an important tool for the conservation of wild animals threatened with extinction. Assisted reproduction procedures in mountain lions, however, are scarce, especially those related to sperm cryopreservation. In this context, this study aimed to evaluate the freezing capacity of semen from adult mountain lions in captivity through the comparison of two concentrations of glycerol in the freezing media. Five adult male mountain lions were used, held at the Rehabilitation Center for Wild Animals of Mato Grosso do Sul (CRAS/MS). Samples were collected by electroejaculation and evaluated for physical appearance, volume, sperm progressive status, sperm motility, sperm concentration and sperm motility index. Each ejaculate was spliced into two aliquots and diluted in Tris-citrate-half egg yolk, at final concentrations of 5 and 7.5% glycerol, cooled at a rate of -0.55ºC/min and frozen at a rate of -5.8ºC/min. Once thawed, the samples were re-evaluated and tested for thermoresistance and hypoosmotic swelling. The suggested protocol for cryopreservation and thawing of semen is efficient in both glycerol concentrations tested, with no difference (p>0.05) between them.

Cell population indexes of spermatogenic yield and testicular sperm reserves in adult jaguars (Panthera onca).

The intrinsic yield of spermatogenesis and supporting capacity of Sertoli cells are the desirable indicators of sperm production in a species. The objective of the present study was to quantify intrinsic yield and the Sertoli cell index in the spermatogenic process and estimate testicular sperm reserves by histological assessment of fragments obtained by testicular biopsy of five adult jaguars in captivity. The testicular fragments were fixed in 4% glutaric aldehyde, dehydrated at increasing alcohol concentrations, included into hydroxyethyl methacrylate, and were cut into 4 microm thickness. In the seminiferous epithelium of the jaguar, 9.2 primary spermatocytes in pre-leptotene were produced by "A" spermatogonia. During the meiotic divisions only 3.2 spermatids were produced by a primary spermatocyte. The general spermatogenic yield of the jaguar was about 23.4 cells and each Sertoli cell was able to maintain about 19.2 germ cells, 11 of them were round spermatids. In each seminiferous epithelium cycle about 166 million spermatozoa were produced by each gram of testicular tissue. In adult jaguars, the general spermatogenic yield was similar to the yield observed in pumas, greater than that observed for the domestic cat, but less compared to most domestic animals.

Morfologia testicular e qualidade espermática de touros da raça Nelore, submetidos à dieta contendo gossipol / Testes morphology and sperm quality of Nelore bulls fed a gossypol-enriched diet

Com o objetivo estudar o efeito de dieta contendo gossipol sobre a qualidade seminal e morfologia dos testículos de touros da raça Nelore, utilizaram-se 16 animais, distribuídos em dois tratamentos: T1, oito touros submetidos a dieta contendo 20 por cento de caroço de algodão e T2, oito touros submetidos a dieta isenta de gossipol. Realizaram-se coletas de sêmen para avaliar a qualidade seminal. Os testículos dos animais foram retirados ao final dos 73 dias de experimento, fragmentos dos mesmos foram preparados para serem analisados em microscopia eletrônica de transmissão e outros fixados em lâminas e corados por hematoxilina-eosina. O consumo médio de caroço de algodão/animal/dia de 1,71kg interferiu na morfologia dos testículos; os túbulos seminíferos apresentaram menor espessura de parede, redução no número de camadas de células espermatogênicas e no volume dos núcleos da célula de Sertoli em relação aos do T2. Por microscopia eletrônica observouse que os touros do T1 apresentavam células de Sertoli com membrana citoplasmática enrugada, acúmulo de lipídeos no citoplasma, mitocôndrias deformadas e algumas espermátides estavam em apoptose. Touros do T1 apresentaram redução na motilidade espermática, aumento no percentual de defeitos espermáticos maiores e totais em relação aos animais do T2. O gossipol alterou a morfologia testicular e reduziu a qualidade espermática dos touros [...]

We studied the effect of a gossypol-enriched diet on the quality of semen and the morphology of testes of Nelore bulls. Sixteen animals were scheduled for 2 treatments: T1, 8 bulls fed a diet containing 20 percent cottonseed and T2, 8 bulls fed a gossypol-free diet. Semen samples were collected in order to evaluate the semen quality. At the end of the 73-day period of treatment, the testes were removed and tissue fragments were prepared for transmission electron microscopy or fixed on slides and stained with haematoxylin-eosin for light microscopy. The mean daily intake of cottonseed by T1 bulls was 1.71 kg/animal, which interfered on testes morphology; the seminiferous tubules had less thick walls, fewer layers of the spermatogenic lineage cells and reduced volumes of Sertoli cell nuclei with regard to T2 bulls. Electron microscopy analyses revealed that the Sertoli cells from T1 animals had a rough cytoplasmic membrane, lipid accumulation in the cytoplasm, deformed mitochondriae; in addition, some apoptotic spermatids were seen. T1 bulls showed reduction of sperm motility and an increase of the percentage of major and total sperm defects in relation to the animals of T2 group. It is concluded that a gossypol-enriched diet promoted alterations in the testicles morphology and it reduced the sperm quality of bulls [...]

Interbular space characterization in adult capybara (Hydrochoerus hydrochaeris) testis

This study aimed at the qualitative and quantitative characterization of the different intertubular tissue components in adult male capybara. Testes of 15 adult capybaras from a commercial slaughterhouse of wild animals were used. The average percentage of Leydig cells was 32.95 ± 10.03 percent, whereas in the other components of the intertubular space it was 14.95 ± 4.79 percent, thus the whole intertubular space totalized 47.91 ± 11.48 percent. The average weight of the testes were 32.83 ± 4.78g, while the animals' body weight were around 52.37 kg and the gonadosomatic and leydigosomatic indexes were 0.125 ± 0.02 percent and 0.0411 ± 0.0148 percent respectively. It was concluded that the leydigosomatic index of the capybaras was lower than the one observed in rats, as well as the arrangement of the components in intertubular tissue was similar to pattern I of Fawcett's classification as was found in guinea-pigs and chinchillas.

Objetivou-se com esta pesquisa caracterizar qualitativamente e quantitativamente os diferentes componentes do tecido intertubular de capivaras machos adultas. Utilizou-se testículos de 15 capivaras adultas, provenientes de um abatedouro comercial de animais silvestres. Encontrou-se um percentual médio de células de Leydig de 32,95+10,03 por cento, dos demais componentes do espaço intertubular de 14,95+4,79 por cento e um total de espaço intertubular de 47,91+11,48 por cento. O peso médio dos testículos foi de 32,83+4,78g, o peso corporal dos animais foi em torno de 52,37kg e os índices gonadossomático e leydigossomático foram, respectivamente, 0,125+0,02 por cento e 0,0411+0,0148 por cento. Concluiu-se que o índice leydigossomático das capivaras é menor que o observado em ratos e que o arranjo dos componentes do tecido intertubular é semelhante ao descrito como padrão tipo I da classificação de Fawcett, similar ao encontrado em cobaios e chinchilas.

The seminiferous epithelium cycle and daily spermatic production in the adult maned wolf (Chrysocyon brachyurus, Illiger, 1811).

The duration of the seminiferous epithelium cycle was estimated in adult maned wolves (Chrysocyon brachyurus, Illiger, 1811), by applying intratesticular injections with tritiated thymidine. The total duration of the seminiferous epithelium cycle in this species was calculated in 8.99 days. So, taking into account that approximately 4.5 cycles of the seminiferous epithelium are necessary for the whole spermatogenesis process to complete, the production of spermatozoa from one spermatogonia will take about 40.45 days. The duration of the spermiogenesis was calculated to be 12.3 days. The eight stages of the seminiferous epithelium cycle were described by the tubular morphology method, which is based either on the form and position of the spermatid nuclei and the occurrence of meiotic divisions. The values of the relative frequency for the pre-meiotic, meiotic and post-meiotic phases in this species were 3.5, 0.78 and 4.8 days, respectively. The maned wolf produces about 29 million spermatozoa a day for each testis gram, therefore being classified among the species provided with a high spermatogenetic efficiency.

Morphometry of leydig cells in the collared peccary (Tayassu tajacu) / Morfometria das células de leydigem catetos (Tayassu tajacu)

This work aimed to determine total and individual volume of Leydigcells, leydigosomatic index and the number of Leydig cells per testisand per gram of testis in the collared peccary (Tayassu tajacu). Testeswere collected from sexually mature collared peccaries, destined forcommercial slaughter. Total and individual volumes of Leydig cellswere 2.02 ml and 1,202.74 x 10-12 ml, respectively. The leydigosomaticindex was 0.022%, and the number of Leydig cell per testis and pergram of testis was 1.7 billion and 92.12 million, respectively. Theseresults show that morphometric characteristics of Leydig cells incollared peccaries are similar to average results observed for most ofthe mammalian species studied.

Objetivou-se com esta pesquisa determinar o volume total e individual das células de Leydig, o índice leydigossomático e o número de células de Leydig por testículo e por grama de testículo em catetos. Utilizaram-se testículos de 10 catetos sexualmente maturos, destinados ao abate comercial. O volume total e individual das células de Leydig foi 2,02ml e 1202,74 x 10-12ml, respectivamente. O núcleo e o citoplasma ocuparam, respectivamente, 12,3% e 87,7% de cada célula de Leydig.O índice leydigossomático foi de 0,022%, enquanto que o número de células de Leydig por testículo e por grama de testículo foi, respectivamente, 1,7 bilhões e 92,12 milhões de células. Concluiu-se que os parâmetros morfométricos estudados para as células de Leydig de catetos estão inseridos na média relatada para a maioria das espécies de mamíferos.

Cat, cougar, and jaguar spermatogenesis: a comparative analysis

This work presents a comparative review about the spermatogenic process in cats (Felis domestica), jaguars (Panthera onca) and cougars (Puma concolor), with emphasis on testicular biometry, gonadossomatic index, volumetric proportion of testicular parenchyma components, tubular diameter, seminiferous epithelial height and seminiferous tubule length. It was an approach of the differences among the cell proportions that allowed conclusions about the overall yield of spermatogenic process and Sertoli cell index in three feline species.

O processo espermatogênico do gato doméstico (Felis domestica), da onça-pintada (Panthera onca) e da onça-parda (Puma concolor) são analisados de forma comparativa, dando-se ênfase à biometria testicular, ao índice gonadossomático, à proporção volumétrica dos constituintes do parênquima testicular, ao diâmetro tubular, à altura do epitélio seminífero e ao comprimento dos túbulos seminíferos. Abordam-se ainda as diferenças entres as razões celulares que permitem conclusões sobre o rendimento do processo espermatogênico e índices de células de Sértoli das três espécies.

Wild boars (Sus scrofa scrofa) seminiferous tubules morphometry

The aim of this data was to analyze morphology and function of the seminiferous tubule in adult wild boars. Testes removed by unilateral castration of five animals were used. The testicular parenchyma was composed by 82.1±2.2 percent of seminiferous tubule and 17.9±2.2 percent of intertubular tissue. The tubular diameter was 249.2±33.0 µm and the seminiferous tubule lenght per gram of testis was 19.3±4.9m. The spermatogonial mitoses efficiency coefficient, meiotic index and spermatogenesis efficiency were 10.34, 2.71 and 30.5 respectively. Each Sertoli cell supported about 13 germinatives cells. The hystometric parameters studied were very similar to those related for domestic boars, however, the wild boars intrinsic efficiency of spermatogenesis and Sertoli cells indexes were smaller than in domestic boars.

Objetivou-se com esta pesquisa estudar as características morfométricas e funcionais dos túbulos seminíferos de javalis adultos. Utilizaram-se testículos de cinco animais submetidos a orquiectomia unilateral. O parênquima testicular foi composto por 82,1 ± 2,2 por cento de túbulos seminíferos e 17,9 ± 2,2 por cento de tecido intertubular. O diâmetro tubular foi de 249,2 ± 33,0µm e o comprimento dos túbulos seminíferos por grama de testículo foi de 19,3 ± 4,9m. O coeficiente de eficiência das mitoses espermatogônias, o rendimento meiótico e o rendimento geral da espermatogênese foram, respectivamente, 10,34, 2,71 e 30,50. Cada célula de Sértoli suportou cerca de 13 células germinativas. Conclui-se que os parâmetros histométricos estudados nesta pesquisa foram muito semelhantes aos valores relatados para suínos domésticos, entretanto, o rendimento intrínseco da espermatogênese e os índices de células de Sértoli de javalis foram relativamente baixos quando comparados com aqueles animais.

Coleta e avaliação do sêmen de catetos (Tayassu tajacu)

Objetivou-se com esta pesquisa testar dois protocolos de eletroejaculação e estudar a motilidade espermática progressiva, vigor, concentração e alterações morfológicas dos espermatozóides de catetos adultos criados em cativeiro. Utilizaram-se seis machos com idade entre 10 e 18 anos. Os animais foram anestesiados com cloridrato de zolazepan e cloridrato de tiletamina e em seguida coletou-se o sêmen com auxílio de um eletroejaculador. O volume médio do ejaculado foi 3,11 + 0,9mL, a motilidade espermática progressiva foi 48,7 ± 31,5 por cento, o vigor, 2,1 + 1,4 e a concentração espermática média, 87,0 + 53,1 x 10(6) sptz/mL. Concluiu-se com esta pesquisa que a utilização de três sessões de 15 estímulos de 12 volts foi suficiente para coleta de sêmen de catetos anestesiados e que os resultados da morfologia espermática da maioria dos catetos trabalhados foram compatíveis com o quadro de degeneração testicular.

Reprodução de capivaras / Reproduction of capybaras / Reproducción de capibaras

A capivara começou a ser explorada no Brasil nos últimos anos, visto sua facilidade de reprodução em cativeiro e alta prolificidade. Tais animais vivem em grupos com um macho dominante, várias fêmeas, indivíduos jovens e sub-adultos. Os machos possuem os testículos localizados subcutaneamente na região inguinal, um dos menores índices gonadossomáticos já registrados em um roedor e uma grande proporção volumétrica de células de Leydig, em comparação a outros animais. As fêmeas chegam à puberdade entre o 10o e 12o mês de idade, a duração do ciclo estral é de 7,5 + 1,2 dias e da gestação, 145 a 150 dias. Apesar de seu grande potencial zootécnico poucas pesquisas têm sido feitas com este animal e muitas perguntas sobre a fisiologia de sua reprodução ainda não foram respondidas.