Morphological and morphometrical changes on adult Wistar rat testis caused by chronic sodium arsenite exposure.

Arsenic is a contaminant that occurs naturally in the environment, and it is related to several diseases, such as cancer and severe metabolic diseases. Sodium arsenite effects on testes rats are not fully understood regarding morphology and stereology; thus, it becomes necessary to evaluate possible changes in these parameters under low concentrations and simulating occupational exposure. Therefore, the aim of this study was to analyze the morphometrical and stereological changes on rat testis treated with sodium arsenite. The treatment was accomplished using 5 mg/kg of sodium arsenite by gastric gavage in Wistar rats, which experiment lasted 8 weeks. Organs were weighed and gonadosomatic index (GSI) was calculated. Using the software Image Pro Plus, seminiferous tubule diameter was measured, and the volume densities of testicular parenchymal components were obtained. It was counted 200 hundred spermatozoa and classified as normal or abnormal. The parameters means of control (N = 5) and treated (N = 7) groups were compared by U Mann-Whitney's test, and the results were considered significant for P < 0.05. We observed a decrease in seminiferous tubule diameter, as well as testis weight. These finds may be related with disorders of testosterone metabolism due to activation of immunological responses of macrophage, which inhibit the steroidogenesis. Thus, we conclude that sodium arsenic does not impair the animal's general health, but its exposure induces biochemical and tissue changes.

Grape juice concentrate alleviates epididymis and sperm damage in cadmium-intoxicated rats.

The possibility of long-term grape juice concentrate (GJC) consumption conferring a protective effect against cadmium (Cd)-induced damage to the epididymis, completely preserving sperm profile, was evaluated here for the first time in the scientific literature. Male Wistar rats (n = 6/per group) received an intraperitoneal Cd injection (1.2 mg/Kg) at age 80 days and GJC (2 g/Kg) by gavage from 50 days until 136 days old. Groups receiving either Cd or GJC were added. An intraperitoneal injection of saline (0.9%) and water by gavage was administered in the absence of treatment with Cd or GJC. Animals were anaesthetized and exsanguinated at 136 days; the vas deferens, left testis and epididymis were removed; and perfusion continued with fixative. The right epididymis was collected for morphological analysis. Cd had a devastating effect demonstrated by reduced sperm count in testes and epididymis, sperm production and normal sperm count, besides increased epididymis sperm transit time and completely disorganized morphology. These alterations were attributed to higher Cd levels in the testes and a lipid peroxidation (LP) process. Consumption of GJC plus Cd intoxication was effective, reducing metal accumulation and LP. Consequently, we could identify a preserved sperm profile, with improvement in testis and epididymis sperm count, normal sperm structure and sperm transit time. Moreover, GJC extends its protective effect to the epididymis, allowing complete re-establishment of its morphology, ensuring successful sperm maturation process. In conclusion, our study indicates long-term GJC as a promising therapy against reproductive chemical intoxication injury damage, preserving sperm prior to ejaculation.

Chronic Intake of Green Propolis Negatively Affecting the Rat Testis.

BACKGROUND: Human and animal evidence suggests that environmental toxicants may have an adverse impact on male reproductive health, reducing the population's reproductive output. Owing to the renewed attraction for natural products, some of them constitute effective alternatives to mitigate these effects. Propolis is a candidate for this use because of its intrinsic properties. In many situations, it improved the testicular damage and alleviated the toxic effects induced by environmental contaminant exposure. OBJECTIVE: The aim of this study was to investigate possible alterations of testicular parameters and certify if its use is really advantageous to the testis, since this could affect rat reproductive function. MATERIALS AND METHODS: Forty-eight adult male Wistar rats were divided into four groups (Co = control, T1 = 3 mg propolis/kg/day, T2 = 6 mg/kg/day, T3 = 10 mg/kg/day) and were exposed during 56 days. The testes were assessed with morphometrical, stereological, and ultrastructural analyses. Cell proliferation and death were diagnosed, respectively, by immunocytochemistry. Connexin 43 (Cx43) and N-cadherin transcript levels were determined by reverse transcription-polymerase chain reaction. RESULTS: Increased cell proliferation and Leydig cell volume were observed in T2, and in contrast, Cx43 upregulation and cell death were observed in T3. Both T2 and T3 showed ultrastructural abnormalities in testicular parenchyma. CONCLUSION: We recommend a cautious intake of propolis to avoid deleterious effects. SUMMARY: Chronic intake of Brazilian green propolis induced N.-cadherin downregulation and decreased on seminiferous tubule volumeIncrease on connexin 43 expression and cell death and decrease in Leydig cell.(LC) number/testis with the concentration of 10 mg/kg/day were observedIncrease on cell proliferation, cytoplasmic proportion, and volume of LC with the concentration of 6 mg/kg/day was detectedThe presence of empty spaces between spermatids and malformed spermatozoa in the lumen of seminiferous tubule was showedThis male reproductive disruption can be linked to phenolic compounds present in Brazilian green propolis. Abbreviation Used: AEC: 3-amino-9-ethylcarbazole; AJ: Adherens junction; AME: Aromadendrin-40-methyl ether; CAPE: Caffeic acid phenethyl ester; Co: Control group; C×43: Connexin 43; DAB: Diaminobenzidine; dNTP: Deoxyribonucleotide phosphate; DSP: Daily sperm production; FA: Ferulic acid; FSH: Follicle-stimulating hormone; GJ: Gap junction; GJIC: Gap junction intercellular communication; HPLC: High-performance liquid chromatography; LC: Leydig cell; LH: Luteinizing hormone; N-cad: N-cadherin; PCNA: Proliferating cell nuclear antigen; PCR: Polymerase chain reaction; RT-PCR: Reverse transcription-polymerase chain reaction; SDM: Standard deviation of mean; T1: Group exposed to 3 mg of propolis/kg/day; T2: Group exposed to 6 mg of propolis/kg/day; T3: Group exposed to 10 mg of propolis/kg/day; TUNEL: Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; WB-ras 2 cells: Ras-transformed rat liver epithelial cell line.

Germinative cells and spermatogenesis of the lizard Tropidurus torquatus (Tropiduridae) from a urban area in the cerrado biome of the brazilian midwest / Células germinativas e espermatogênese do lagarto Tropidurus torquatus (Tropiduridae) de uma área urbana no bioma cerrado do centro-oeste brasileiro

Tropidurus comprises a Neotropical genus of lizard that currently has about 30 species widely distributed in the South American. Among these species, Tropidurus torquatus, which has the characteristic of great physiological plasticity, occupying a variety of habitats in open areas and urbanized environments. Considering this, the aim of the study was to investigate the germinative cells and spermatogenesis of a population of T. torquatus in an urban area under Cerrado Biome influences to understand how to establish the temporal development of germinative cells and spermatogenesis during a period of one year. Individuals were obtained in the Zoological Collection of Vertebrates at the Universidade Federal de Mato Grosso (UFMT), and the germinative cells and full spermatogenesis were described with light microscopy. Tropidurus torquatus presented germ cells with similar characteristics already documented for the other species of lizards and reptiles. Spermatogonia type A and B, primary and secondary spermatocytes, and spermatids were present in almost all months evaluated. The gonadosomatic ratio presented its highest value in October, moment in which spermatogenesis presented all the germinative cells and spermatozoa in the lumen, of the seminiferous tubules. In the seasonal climate of the Cerrado Biome, we observe discontinuous spermatogenesis in T. torquatus with the production of spermatozoa in almost every month of the year, however with sperm storage in the epididymis during the phase of testicular regression.

O gênero de lagartos neotropicais Tropidurus tem cerca de 30 espécies amplamente distribuídas no continente sul-americano. Entre essas espécies, Tropidurus torquatus é a que apresenta grande plasticidade fisiológica, ocupando uma variedade de hábitats em áreas abertas e ambientes urbanizados. Considerando isso, o objetivo do estudo foi investigar as células germinativas e a espermatogênese de uma população de T. torquatus em uma área urbana sob a influência do Bioma Cerrado para entender como é estabelecido o desenvolvimento temporal das células germinativas e a espermatogênese durante o período de um ano. Indivíduos foram obtidos na Coleção Zoológica de Vertebrados da Universidade Federal de Mato Grosso (UFMT). A caracterização das células germinativas e da espermatogênese foi realizada através de microscopia de luz. Tropidurus torquatus apresentou células germinativas com características similares às já documentadas para outras espécies de lagartos e répteis. Espermatogônias do tipo A e B, espermatócitos primários e secundários e espermátides estiveram presentes praticamente em todos os meses. A razão gonadossomática apresentou seu valor máximo em outubro, momento em que a espermatogênese apresentou todas as células germinativas e espermatozoides no lúmen do túbulo seminífero. No clima sazonal do Bioma Cerrado observamos espermatogênese descontínua em T. torquatus com a produção de espermatozoides em quase todos os meses do ano, porém com estocagem nos epidídimos durante a fase de regressão testicular.

Sperm morphology of Muscidifurax uniraptor (Hymenoptera: Chalcidoidea: Pteromalidae).

Sperm morphology of the parasitoid Muscidifurax uniraptor was investigated under light and transmission electron microscopy. M. uniraptor sperm are filiform, spiraled, approximately 150 µm in length, with a distinctive head, hooded by an extracellular sheath and a flagellum. This extracellular layer, from which many filaments radiate, measures approximately 90 nm in thickness and covers a small acrosome and the anterior nuclear region. The acrosome is composed of an acrosomal vesicle and a perforatorium with its base inserted in the nuclear tip. The nucleus is filled with homogeneously compacted chromatin. The centriolar adjunct extends towards the anterior portion in a spiral around the nucleus for 3.5 µm in length. The two mitochondrial derivatives begin exactly at the centriole adjunct base and, in cross-section, have a circular shape with equal areas that are smaller than the axoneme diameter. It is coiled, with 9 + 9 + 2 microtubules and begins from the centriole, just below the nuclear base. The axoneme is connected to the mitochondrial derivatives by two small irregularly shaped masses. Between the derivatives and the axoneme, the 'center-flagellar material' is observed. Overall, these characteristics are recognized in other Chalcidoidea, especially in the eurytomids, but together they form a set of species-specific data.

Grape juice concentrate (G8000(®) ) intake mitigates testicular morphological and ultrastructural damage following cadmium intoxication.

Cadmium is a well-known testicular toxicant, and parts of the world population are exposed chronically by inhalation or by food and water intake. Grape products have been highlighted as important sources of bioactive compounds, having anti-inflammatory, anti-oxidant and metal chelating properties. Since maintenance of tissue morphology is essential for testicular sperm development and hence male fertility, we analysed the protective effect of grape juice concentrate (GJC) (G8000(®) ) consumption on testicular morphology in rats exposed to cadmium. Thus, four groups of male Wistar rats (n = 6 per group), 50 days old, ingested either water or G8000(®) (2 g/kg/day) until they had completed one spermatogenic cycle in adult life (136 days old). Cadmium (1.2 mg / kg) was injected intraperitoneally when the animals were 80 days old into one of the water and one of the G8000 groups; intraperitoneal saline was used as a control in the other two groups. Animals anaesthetised and exsanguinated at 136 days and then perfused with Karnovsky's fixative and then the testes were collected for morphological analysis. We describe evident disruption of testicular morphology by cadmium, with alteration in tissue component proportions, reduced Leydig cells volume and initial signs of an inflammatory process. Ultrastructural analysis showed greater damage, suggesting spermatogenesis disruption. G8000(®) ingestion allowed tissue architecture to be re-established, as was corroborated by our stereological and morphometric findings. Animals from the group where G8000(®) had been administered together with cadmium revealed a significant reduction in macrophages and blood vessel volume, suggesting diminished inflammation, when compared to animals that received only cadmium. Moreover, smaller number of ultrastructural alterations was noted, revealing fewer areas of degeneration and disorganized interstitium. In conclusion, our results demonstrate that GJC consumption prevented the spermatogenic disruption promoted by cadmium, and thus could be a promising form of therapy against male infertility.

Toxicological evaluation of ametryn effects in Wistar rats.

São Paulo state, Brazil, is one of the main areas of sugar cane planting in the world. Extensive use of ametryn, a triazine herbicide, in sugar cane agriculture and the properties of this herbicide suggest it could be present in the environment as a potential contaminant of soil, surface water, groundwater, and river sediment. In order to clarify the mechanism through which ametryn could be toxic, an in vivo study with Wistar rats was conducted using hematological, biochemical, molecular, morphological and genotoxic approaches. For this purpose, two sub-lethal ametryn concentrations (15 mg and 30 mg/kg/day) were administered to 42 rats divided into three groups (n=12) by gavage during 56 days, whereupon blood, liver and bone marrow were collected. The results showed ametryn genotoxic activity by in vivo micronuclei testing. This event probably occurred as consequence of oxidative stress induction demonstrated by GSTM1 transcript levels increase (indicating complexation between ametryn and/or metabolites with GSH) and by SOD activity decrease. Also, Mn-SOD transcripts were increased, probably avoiding mtDNA damage caused by EROS. These mechanisms displayed hepatic stellate cell (HSCs) activation because two major biomarkers were regulated, connexin and cadherin. N-cad transcripts were increased on both exposed groups while E-cad decreased in the T1 group, indicating epithelial-to-mesenchymal transition. In addition, Cx43 transcripts were decreased suggesting an increase in collagen content. Volumetric proportion of sinusoids was significantly decreased in T1 group and no significant alteration in hepatocyte volume was observed, indicating an increase in the space of Disse, due to fibrosis. Hepatocyte nuclei showed significant decrease in diameter and volume. Few hematological alterations were found. We emphasize the importance of other approaches, such as cell death and proliferation assays, so that ametryn toxicity can better be understood.

The effects of cyclosporin A and Heteropterys tomentosa on the rat liver.

Cyclosporin A (CsA) is a widely employed immunosuppressive drug that is associated with several side effects, among then hepatotoxicity. Heteropterys tomentosa is a Brazilian plant efficient in reducing damage caused by CsA on the rat testis and prostate. The aim of this study was to evaluate the effect of CsA and H. tomentosa (administered isolated or simultaneously) on the liver of Wistar rats. The animals were treated daily with water (control), CsA (15 mg/kg/day), H. tomentosa infusion or CsA+H. tomentosa, for 21 or 56 days. The treatments did not alter liver morphology or cause fibrosis. H. tomentosa administered for 21 days increased the number of hepatocyte nuclei and Kupffer cell volumetric proportion. After 56 days of treatment, H. tomentosa administration did not alter the parameters analyzed. Biochemical plasma dosages and liver stereology showed impairment caused by CsA-treatment after 21 days; these results were not observed after 56 days of treatment. The simultaneous treatment with CsA and H. tomentosa for 21 or 56 days did not alleviate nor accentuate CsA hepatic effects. The present study showed that the 21 days treatment with CsA caused more alteration to the liver than the 56 days treatment; this could be related to hepatic recovery after the long term treatment.

Hepatoprotective effect of Arctium lappa root extract on cadmium toxicity in adult Wistar rats.

This study was performed to determine the effects of Arctium lappa (Al) to protect against cadmium damage in the rat liver. Male rats received a single i.p. dose of CdCl2 (1.2 mg/kg body weight (BW)) with or without Al extract administered daily by gavage (300 mg/kg BW) for 7 or 56 days. After 7 days, Al caused plasma transaminase activity to diminish in groups Al (glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT)) and CdAl (GPT). After 56 days, GOT and GPT plasma activities were reduced in the Cd group. No alteration in plasma levels of creatinine, total bilirubin, and total protein were observed. GOT liver activity increased in the Cd group. No alteration was observed in superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), and malondialdehyde (MDA) dosage. In the Cd group, hepatocyte proportion decreased and sinusoid capillary proportion increased. In the Al and CdAl groups, the nuclear proportion increased and the cytoplasmic proportion decreased. The hepatocyte nucleus density reduced in Cd and increased in the Al group. After 56 days, there was no alteration in the Cd group. In Al and CdAl groups, the nuclear proportion increased without cytoplasmic proportion variation, but the sinusoid capillary proportion was reduced. The hepatocyte nucleus density decreased in the Cd group and increased in the Al and CdAl groups. In conclusion, the liver function indicators showed that A. lappa protected the liver against cadmium toxicity damage.

Colon cancer and swimming exercise: effect on wistar rat testes

This study was undertaken to determine whether colon cancer (CC) and chronic swimming exercise alter rat testis. Eleven weeks old rats were distributed into control group (n=6) and the groups that were induced to develop CC by dimethylhydrazine injections (nEG, EG0, EG2 and EG4; n=10 each group). In the group nEG, the rats did not swim, whereas groups EG0, EG2 and EG4, underwent a swimming program with distinct loads (0, 2 and 4% of body mass, respectively) for 35 weeks. The morphometry, stereology and cell counts showed damage caused by the CC on the germ epithelium. These results were noteworthy since this was the first report to associate the CC with testicular damage. Swimming exercise had no significant role in reducing, or increasing the CC effects on the testis, despite having slightly improved the testis structure of the exercised rats without load. In conclusion, CC caused testis impairment, which could not be avoided by the swimming exercise.

Heteropterys tomentosa (A. Juss.) infusion counteracts Cyclosporin a side effects on the ventral prostate.

BACKGROUND: Cyclosporin A (CsA) is an immunosuppressive drug widely used in treatment of auto-immune diseases or after organ transplants. However, several side effects are commonly associated with CsA long term intake, some regarding to loss of reproductive organ function due to oxidative damage. Considering that phytotherapy is an important tool often used against oxidative stress, we would like to describe the beneficial effects of Heteropterys tomentosa intake to minimize the damage caused by CsA to the ventral prostate tissue of Wistar rats under laboratorial conditions. METHODS: Thirty adult Wistar rats (Rattus norvegicus albinus) were divided into: control group (water); CsA group (Cyclosporin A); Ht group (H. tomentosa infusion) and CsA + Ht group (CsA and H. tomentosa infusion). Plasmic levels of hepatotoxicity markers, triglycerides, cholesterol and glucose were quantified. The ventral prostate tissue was analyzed under light microscopy, using stereological, morphometrical and immunohistochemical techniques. RESULTS: H. tomentosa did not cause any alterations either of the plasmic parameters or of the ventral prostate structure. CsA caused alterations of GOT, total and indirect bilirubin, cholesterol, triglycerides and glucose levels in the plasma; CsA-treated rats showed alterations of the ventral prostate tissue. There were no alterations regarding the plasma levels of GOT, triglycerides and glucose of CsA + Ht animals. The same group also showed normalization of most of the parameters analyzed on the ventral prostate tissue when compared to the CsA group. The treatments did not alter the pattern of AR expression or the apoptotic index of the ventral prostate epithelium. CONCLUSIONS: The results suggest a protective action of the H. tomentosa infusion against the side effects of CsA on the ventral prostate tissue, which could also be observed with plasmic biochemical parameters.

Advantage of Guaraná (Paullinia cupana Mart.) supplementation on cadmium-induced damages in testis of adult Wistar rats.

Paullinia cupana is an Amazonian bush whose seeds have long been used in folk medicine. However, most of the therapeutic properties attributed to this plant are broad and nonspecific, although an antioxidant activity has been reported.  On the other hand, cadmium is a heavy metal known for increasing free radicals, hence resulting in cellular oxidative damages. This study was designed to evaluate whether Paullinia cupana is able to reduce cadmium-induced morphological impairment in Wistar rat testis. Adult male Wistar rats 110 days old were ip injected with cadmium (1.15 mg/kg BW [body weight]) and subsequently treated with P. cupana during 56 days.  Furthermore, groups receiving either P. cupana extract or cadmium are mentioned. After the treatment period, testis samples were subjected to histological and stereological analyses. Moderate to severe testicular impairments were shown by the animals exposed to cadmium. However, the animals supplemented with P. cupana after cadmium exposure showed a significant decrease in the proportion of damaged seminiferous tubules. Also, P. cupana supplementation was effective in maintaining the number of Leydig cells per testis in the animals exposed to cadmium. In conclusion, P. cupana supplementation was partially efficient in preventing cadmium from damaging the testis of adult Wistar rats.

Morphology of spermatozoa of Dissomphalus connubialis (Ashmead, 1893) (Hymenoptera: Bethylidae).

In this study, the morphology of spermatozoa of Bethylidae Dissomphalus connubialis (Pristocerinae) was analyzed using light and transmission electron microscopy. Spermatozoa of this species are thin, measure approximately 130 µm in length and comprise a head region and flagellum region. The head is formed by the acrosome and nucleus. The acrosome consists of the acrosome vesicle and the perforatorium, the posterior portion of which is inserted into a cavity at the anterior extremity of the nucleus. The nucleus is compact, electron-dense and measures 15 µm in length. The flagellum possesses two mitochondrial derivatives, two accessory bodies and one axoneme with a 9+9+2 microtubular pattern. The nucleus is connected to the flagellum by the centriole adjunct. Mitochondrial derivatives are compact, apparently without paracrystalline material and with rare mitochondrial cristae. They are asymmetric in length, such that the larger mitochondrial derivative begins parallel to the posterior region of the nucleus and the smaller mitochondrial derivative begins just below the centriole adjunct. The basic structure of spermatozoa of D. connubialis is similar to that of other Aculeata studied. However, this species shows characteristics not seen in other Hymenoptera, such as the wide electron-lucid region that separates the acrosomal vesicle from the perforatorium and the depth of the cavity in the anterior extremity of the nucleus, into which the base of the perforatorium is inserted. There are also characteristics that distinguish this species from Bethylidae Prorops nasuta, including the fact that one of the mitochondrial derivatives lies to parallel to the nucleus over a long distance, the small quantity of cristae, the absence of paracrystalline material in these organelles, and the fact that the accessory microtubules are the first to terminate in the final portion of the flagellum.

Morphology of the epidermis of the neotropical catfish Pimelodella lateristriga (Lichtenstein, 1823) with emphasis in club cells.

The epidermis of Ostariophysi fish is composed of 4 main cell types: epidermal cells (or filament containing cells), mucous cells, granular cells and club cells. The morphological analysis of the epidermis of the catfish Pimelodella lateristriga revealed the presence of only two types of cells: epidermal and club cells. The latter were evident in the middle layer of the epidermis, being the largest cells within the epithelium. Few organelles were located in the perinuclear region, while the rest of the cytoplasm was filled with a non-vesicular fibrillar substance. Club cells contained two irregular nuclei with evident nucleoli and high compacted peripheral chromatin. Histochemical analysis detected prevalence of protein within the cytoplasm other than carbohydrates, which were absent. These characteristics are similar to those described to most Ostariophysi studied so far. On the other hand, the epidermal cells differ from what is found in the literature. The present study described three distinct types, as follows: superficial, abundant and dense cells. Differences among them were restricted to their cytoplasm and nucleus morphology. Mucous cells were found in all Ostariophysi studied so far, although they were absent in P. lateristriga, along with granular cells, also typical of other catfish epidermis. The preset study corroborates the observations on club cells' morphology in Siluriformes specimens, and shows important differences in epidermis composition and cell structure of P. lateristriga regarding the literature data.

Morphology of the male reproductive system and spermatozoa in Centris Fabricius, 1804 (Hymenoptera: Apidae, Centridini).

The genera Centris and Epicharis constitute the Centridini and are widely distributed in the Neotropical region. Centris is also found in the southern portion of the Neartic region, although both genera are more abundant in the humid tropical regions. To describe the structure of the male reproductive system and spermatozoa, light and transmission electron microscopy were used. The male reproductive system of Centris sp. is formed by a pair of testes, a pair of deferent ducts, a pair of seminal vesicles, a pair of accessory glands and an ejaculatory duct connected to the external genitalia, the aedeagus. In this species, testes and the pre-vesicular deferens ducts as well as the seminal vesicles are encapsulated in a single conjunctive capsule, the scrotal membrane. Each testis consists of four testicular follicles, made up of cysts with up to 64 germinative cells. Histologically, the seminal vesicles are formed by a simple cylindrical epithelium, basal membrane and muscular tunic. The spermatozoa of Centris analis, C. fuscata, C. tarsata and Centris sp. are morphologically similar. They have two easily distinguishable regions: the head and flagellum. The head is formed by the two-layer acrosome, the linear nucleus and the flagellum, the centriole adjunct, the axoneme of pattern 9+9+2 microtubules, two asymmetric mitochondrial derivatives and two accessory bodies. These Centris species share various morphological characteristics of the male reproductive system and spermatozoa with the other bees previously described, indicating that several characteristics are synapomorphic for the family Apidae. Studies on the morphology of the male reproductive system and spermatozoa in Hymenoptera have demonstrated the diversity of the information provided by these reproductive structures, which can be used in taxonomy studies and the phylogeny of this important group of insects.

The descriptions of new microanatomical structures of the male reproductive system and sperm of Myschocyttarus cassununga (Hymenoptera: Vespidae).

The male reproductive system of Mischocyttarus cassununga consists of two testes, each one with three follicles, as occurs in most Vespidae. The seminal vesicle is divided in two chambers, separated by a constriction, so that the anterior locus is a little larger. In the testicular follicles, the spermatozoa are organized in cysts, with approximately 128 per cyst, where the nuclei are oriented toward the follicle center. The spermatozoa of M. cassununga is about 97 µm in length, which makes them the shortest sperm described for Vespidae. Sperm ultrastructure of M. cassununga is very similar of the others Vespidae. But, despite these similarities, the bilobated mitochondrial derivative represents an autapomorphy for M. cassununga. The subdivision of the seminal vesicle has never been observed in any other Vespidae. Thus, this study supports the validity of insect sperm morphology as a tool for phylogenetic analysis within Hymenoptera.

Histomorphometric evaluation of the neotropical brown brocket deer Mazama gouazoubira testis, with an emphasis on cell population indexes of spermatogenic yield.

Information on the reproductive biology of neotropical cervids is scarce. Therefore, the aim of this study was to perform biometric, histologic and stereologic analyses of the brown brocket deer Mazama gouazoubira testis, with an emphasis on the intrinsic yield and the Sertoli cell index. Seven adult males kept in captivity were used. The animals were immobilized; anesthetized and testicle fragments were obtained by biopsy incision. The material was fixed, processed and examined by routine histological methods for light microscopy. The average body weight was 17.2kg, from which 0.40% were allocated in gonads and 0.33% in seminiferous tubules, which represented 85.9% of the testis parenchyma. The mean albuginea width and volume were 345.7µm and 3.5mL (5.3% of the testicular weight), respectively. The mean mediastinum volume of both testicles was 1.0mL (1.5% of the testicular weight) and the testicular parenchyma volume corresponded to 93.1% of total testicular weight (64.9g). The seminiferous tubules diameter was 224.4µm, while the epithelium height was 69.6µm. On average, an adult brown brocket deer showed a total of 1418m of seminiferous tubules in both testicles (21.5m per gram of testis). Each stage I seminiferous tubular cross section contained 1.10 type A spermatogonia, 13.4 primary spermatocytes in pre-leptotene/leptotene, 13.7 spermatocytes in pachytene, 48.8 round spermatids and 3.7 Sertoli cells. The general yield of spermatogenesis was 44.7 cells and the Sertoli cell index was 13.2. The qualitative and quantitative description of testicular histology of brown brocket deer help to understand its spermatogenic process and to establish parameters for the reproductive biology of this wild species. Furthermore, the data from the present research will help further studies using other species of Brazilian cervids, especially endangered ones, making an additional effort to the species preservation.

More resistant tendons obtained from the association of Heteropterys aphrodisiaca and endurance training.

BACKGROUND: Popular Brazilian medicine uses Heteropterys aphrodisiaca infusion as a tonic or stimulant, for the treatment of nervous debility and breakdown and for muscle and bone weakness. This study investigated the effects of Heteropterys aphrodisiaca infusion on the tendon properties and extracellular matrix of rats under endurance training. METHODS: Wistar rats were grouped as follows: CS- control sedentary, HS- H. aphrodisiaca sedentary, CT-control trained, HT- H. aphrodisiaca trained. The training protocol consisted in running on a motorized treadmill, five times a week, with weekly increase in treadmill speed and duration. Control groups received water while the HS and HT groups received H. aphrodisiaca infusion, daily, by gavage for the 8 weeks of training. Achilles tendons were frozen for biochemical and biomechanical analysis or preserved in Karnovsky's fixative, then processed for histomorphological analysis with light microscopy. RESULTS: Biomechanical analysis showed significant increase in maximum load, maximum stress, modulus of elasticity and stiffness of the HT animals' tendons. The metalloproteinase-2 activity was reduced in the HT group. The compression region of HT animals' tendons had a stronger and more intense metachromasy, which suggests an increase in glycosaminoglycan concentration in this region of the tendon. The most intense birefringence was observed in both compression and tension regions of HT animals' tendons, which may indicate a higher organizational level of collagen bundles. The hydroxyproline content increased in the HT group. CONCLUSIONS: The association of endurance training with H. aphrodisiaca resulted in more organized collagen bundles and more resistant tendons to support higher loads from intense muscle contraction. Despite the clear anabolic effects of Heteropterys aphrodisiaca and the endurance exercise association, no side effects were observed, such as those found for synthetic anabolic androgenic steroids.

Antioxidative and in vitro antiproliferative activity of Arctium lappa root extracts.

BACKGROUND: Arctium lappa, known as burdock, is widely used in popular medicine for hypertension, gout, hepatitis and other inflammatory disorders. Pharmacological studies indicated that burdock roots have hepatoprotective, anti-inflammatory, free radical scavenging and antiproliferative activities. The aim of this study was to evaluate total phenolic content, radical scavenging activity by DPPH and in vitro antiproliferative activity of different A. lappa root extracts. METHODS: Hot and room temperature dichloromethanic, ethanolic and aqueous extracts; hydroethanolic and total aqueous extract of A. lappa roots were investigated regarding radical scavenging activity by DPPH, total phenolic content by Folin-Ciocalteau method and antiproliferative in vitro activity was evaluated in human cancer cell lines. The hydroethanolic extract analyzed by high-resolution electrospray ionization mass spectroscopy. RESULTS: Higher radical scavenging activity was found for the hydroethanolic extract. The higher phenolic contents were found for the dichloromethane, obtained both by Soxhlet and maceration extraction and hydroethanolic extracts. The HRESI-MS demonstrated the presence of arctigenin, quercetin, chlorogenic acid and caffeic acid compounds, which were identified by comparison with previous data. The dichloromethane extracts were the only extracts that exhibited activity against cancer cell lines, especially for K562, MCF-7 and 786-0 cell lines. CONCLUSIONS: The hydroethanolic extracts exhibited the strongest free radical scavenging activity, while the highest phenolic content was observed in Soxhlet extraction. Moreover, the dichloromethanic extracts showed selective antiproliferative activity against K562, MCF-7 and 786-0 human cancer cell lines.

Testicular histomorphometry and ultrastructure of rats treated with cadmium and Ginkgo biloba.

The aim of this study is to investigate the association of a single low dose of Cd and daily doses of Ginkgo biloba extract (GbE) on the testis and accessory glands of rats. The animals were treated with a single dose of 3 µmol/kg body weight of cadmium chloride (CdCl2) and/or 100 mg/kg body weight of GbE. The plasma testosterone levels; corporal, testicular, and accessory glands weight; gonadosomatic index, volumetric proportion; and absolute volume of testicular components did not change after the treatments. CdCl2 caused significant reduction in Leydig cells volume and altered Leydig cell morphology, as well as vacuolated Sertoli cells cytoplasm, irregular chromatin condensation of late spermatids, and modified acrosome formation. However, animals that received GbE did not show these alterations. The reversal of Cd-induced alterations by the extract is a strong indication that G. biloba is helpful in diminishing the effect of Cd toxicity.