Could vitamin C and zinc chloride protect the germ cells against sodium arsenite?

Arsenic (As) is commonly associated with natural and human processes such as volcanic emissions, mining and herbicides production, being an important pollutant. Several studies have associated As intake with male fertility reduction, thus the aim of the present study was to evaluate whether vitamin C and/or zinc would counteract As side effects within the testicles. Adult male Wistar rats were divided into six experimental groups: control, sodium arsenite (5 mg/kg/day), vitamin C (100 mg/kg/day), zinc chloride (ZnCl2; 20 mg/kg/day), sodium arsenite + vitamin C and sodium arsenite + ZnCl2. Testicles and epididymis were harvested and either frozen or routinely processed to be embedded in glycol methacrylate resin. As reduced the seminiferous epithelium and tubules diameter due to germ cell loss. In addition, both the round spermatids population and the daily sperm production were reduced. However, ZnCl2 and vitamin C showed to be effective against such side effects, mainly regarding to sperm morphology. Long-term As intake increased the proportions of abnormal sperm, whereas the concomitant intake of As with zinc or vitamin C enhanced the proportions of normal sperm, showing that such compounds could be used to protect this cell type against morphological defects.

Effects of Arctium lappa on Cadmium-Induced Damage to the Testis and Epididymis of Adult Wistar Rats.

The protective role of Arctium lappa (AL) on the testes of rats acutely exposed to cadmium (Cd) was tested. The rats were randomly divided into a control group (C-group) and three major experimental groups, which were further subdivided into minor groups (n = 6) according to the experimental period (7 or 56 days). The C-group was subdivided into C-7 and C-56 [receiving a single saline solution, intraperitoneal (i.p.), on the first day]; the AL-group, AL-7, and AL-56, received AL extract (300 mg/kg/daily); the Cd group, Cd-7 and Cd-56, received a single i.p. dose of CdCl2 (1.2 mg/kg body weight (BW)) on the first day; the CdAL group, CdAL-7 and CdAL-56, received the same Cd dose, followed by AL extract. Water or AL extract was administered daily by gavage. After either 7 or 56 days, the testis and accessory glands were removed after whole-body perfusion. Exposure to Cd and CdAL decreased the weight of the testis and epididymis, the gonadosomatic index, seminiferous tubular (ST) diameter, and ST volumetric proportion, and increased the volumetric proportion of interstitium after 56 days. In the epididymis caput, the tubular volumetric proportion decreased along with an increase of interstitial volumetric proportion and epithelium height after 56 days. The alterations observed were less severe only after 7 days. A progressive testicular damage resulted mainly in tubules lined only by Sertoli cells. The sperm number and cell debris decreased in the epididymis. We demonstrated that the testicular damage induced by single acute i.p. exposure to Cd occurred despite the daily oral intake of AL extract.

Leydig cell number and sperm production decrease induced by chronic ametryn exposure: a negative impact on animal reproductive health.

Ametryn is an herbicide used to control broadleaf and grass weeds and its acute and chronic toxicity is expected to be low. Since toxicological data on ametryn is scarce, the aim of this study was to evaluate rat reproductive toxicity. Thirty-six adult male Wistar rats (90 days) were divided into three groups: Co (control) and T1 and T2 exposed to 15 and 30 mg/kg/day of ametryn, respectively, for 56 days. Testicular analysis demonstrated that ametryn decreased sperm number per testis, daily sperm production, and Leydig cell number in both treated groups, although little perceptible morphological change has been observed in seminiferous tubule structure. Lipid peroxidation was higher in group T2, catalase activity decreased in T1 group, superoxide dismutase activity diminished, and a smaller number of sulphydryl groups of total proteins were verified in both exposed groups, suggesting oxidative stress. These results showed negative ametryn influence on the testes and can compromise animal reproductive performance and survival.

Effect of the Arrabidaea chica extract on collagen fiber organization during healing of partially transected tendon.

AIMS: After undergoing lesions, tendons have disorganized collagen fibers compared to undamaged tendons. Arrabidaea chica leaves have the aglycones carajurin and carajurone, components of the antocyanins, with a strong pharmacological potential due to their healing properties. Thus, the aim of this study was to investigate the effect of topical application of A. chica extract during tendon healing. MAIN METHODS: The calcaneal tendon of Wistar rats was partially transected with subsequent treatment with A. chica extract (2.13 g/mL) followed by excision on the 7th, 14th and 21st days. Control rats received only saline treatment. KEY FINDINGS: Transmission electron microscopy analysis showed the presence of a large amount of small segments of collagen fibrils in the transected region of the tendons on the 7th day in both the control and plant-treated groups. Considering the organization of the collagen fibers, higher values of birefringence were observed under polarization microscopy in the tendons of the plant-treated group on the 14th day compared to the control group. A larger quantity of dermatan sulfate was also detected after plant treatment in the same period. However, lesser dermatan and chondroitin sulfate were detected in the plant-treated group than in the control group on the 21st day. No differences were found in the values of birefringence between these groups. Intense metachromasy was observed in both transected groups on the 21st day. SIGNIFICANCE: In conclusion, the use of A. chica extract improves collagen organization and increases the quantity of dermatan sulfate on the 14th day of the tendon healing.

Cyclosporin A causes impairment of the ventral prostate tissue structure of Wistar rats.

Cyclosporin A (CsA) is an immunosuppressive drug widely used in medicine to reduce the immune system activity and, therefore, the risk of organ rejection after transplantation. However, many side effects can be related to its use, such as, reduction in serum testosterone levels due to damage of the testis structure and, consequently, male infertility. The present study aims to evaluate the effects of chronic CsA administration on the ventral prostate tissue (15 mg/kg per d, for 56 days). Stereological, morphometrical, morphological and ultrastructural observations were employed. The plasmatic testosterone and glucose levels were measured. An androgen receptor (AR) immunohistochemical method was applied on ventral prostate sections. Apoptosis was detected with the terminal deoxynucleotidyl transferase dUTP nick end labeling technique. CsA treatment caused reduction in plasmatic testosterone levels and an increase in glycemia. The volume of all ventral prostate tissue components (lumen, epithelium and muscular and nonmuscular stroma) and ventral prostate weight were reduced in the CsA-treated group. Light and transmission electron microscopy confirmed epithelium atrophy of treated animals. There was no alteration of AR expression or apoptotic index. CsA chronic treatment in the therapeutic doses caused damage to prostate tissue of adult Wistar rats, probably due to increase in the glucose levels and reduction in the plasmatic testosterone levels.

Morphology of male reproductive systems in Ephemeroptera: intrinsic musculature.

Although the Ephemeroptera have been studied over a long period of time, there are still few studies on the morphology of male reproductive system. The spermatic ducts are considered conserved among different Ephemeroptera groups. However, previous studies distinguished different organizational patterns of the spermatic duct intrinsic musculature. This study describes the morphology of the spermatic ducts, especially their musculature, in six species of Ephemeroptera, representing five families. We have observed variations in the organizational pattern of the spermatic ducts, even between species from the same family. Moreover, all species studied had intrinsic musculature in the spermatic ducts although with different organizational patterns. Thus, we believe that this musculature is important to move the spermatozoa along the ducts of all Ephemeroptera and not only of those with aflagellated spermatozoa (Leptophlebiidae). The variations in musculature organization must be related to differences in reproductive physiology (i.e., copula duration) and not only with spermatozoa characteristics.

Morphometry of the midgut of Melipona quadrifasciata anthidioides (Lepeletier) (Hymenoptera: Apidae) during metamorphosis

In Hymenoptera, midgut changes begin in the last instar. At this stage, the larval epithelial digestive cells degenerate, leaving only the basal membrane and the regenerative cells which will develop into a new epithelium during the pupal stage and in the adult. Epithelium renewal is followed by changes in volume and shape of the midgut. Morphometric analysis of digestive cells and total midgut volume of Melipona quadrifasciata anthidioides (Lepeletier) were conducted to verify whether cell volume increase are sufficient to account for the total midgut volume increase that occurs during metamorphosis. An increase in midgut volume was verified in spite of the scarcity of cell proliferation found during metamorphosis. At the end of metamorphosis, the increase in cell volume was not sufficient to explain the increase in volume of the midgut, indicating that an increase in the number of digestive cells is apparently necessary. Nevertheless, the mechanism by which regenerative cells reconstitute the epithelium during metamorphosis remains unknown.

Does heteropterys aphrodisiaca administration andendurance training alter bones of mature rats?

Heteropterys aphrodisiaca infusion, alone or associated with endurance training, was investigated in rat bonesin relation to their mechanical properties, collagen content and morphology. Male rats were divided into fourgroups (n = 8): CS- control sedentary, HS- H. aphrodisiaca sedentary, CT–control trained, HT‑H. aphrodisiacatrained. The training protocol consisted in running on a motorized treadmill, 5 times a week, for 8 weeks,with weekly increase in treadmill velocity and duration. Control groups received water while HS and HTgroups received H. aphrodisiaca infusion (104 mg/animal) by gavage during the 8 weeks. Tibiae werefrozen for collagen dosage and biomechanical analysis or preserved in Karnovsky’s fixative, then processedfor histomorphological analysis by conventional light microscopy and scanning electron microscopy. TheHT group showed significantly higher yield load and yield stress in the tibiae three-point bending test. Themaximum load, stiffness, maximum stress and elastic modulus were statistically similar for the experimentalgroups. The hydroxyproline content, morphometrical and stereological data were not significantly differentfor the four groups. Scanning electron microscopy showed more lacunae and Havers canals in the bone oftrained animals, moreover the osteons were more disorganized, when compared with sedentary groups. Thesealterations may indicate that the bone of trained animals was being remodeled. However, after 8 weeks oftraining, it was not possible verify alterations in morphometrical measurements, collagen content, stiffness andmodulus of elasticity of the trained and treated animals.

Characteristics of the male reproductive system and spermatozoa of leptophlebiidae (ephemeroptera).

This study describes morphological changes in the male reproductive system of Miroculis amazonicus (Savage & Peters) from mature nymphs to subimago stages. The sperm ultrastructure of Massartela brieni (Lestage), Farrodes carioca (Domínguez et al) and Miroculis mourei (Savage & Peters), as well as aspects of cell fragments observed in these species' subimagos deferent ducts were described. Sperm from the three species studied are aflagellated and immotile, while those from F. carioca and Ma. brieni are approximately spherical with a homogenous nucleus and acrosome. Sperm of F. carioca present two or three mitochondria located between the nucleus and the acrosome. In Ma. brieni, only one lateral mitochondria was found. Sperm from Mi. mourei are shaped as a number 'eight', with electron lucent spots inside the nucleus and two mitochondria above the acrosome. Large cell fragments containing degenerative vesicles and some sperm were observed in the deferent duct lumen of the three species. Testes of Mi. amazonicus are extremely reduced in the subimago stage, which suggests that these cell fragments originated from testes degeneration.

Characteristics of the Male Reproductive System and Spermatozoa of Leptophlebiidae (Ephemeroptera)

This study describes morphological changes in the male reproductive system of Miroculis amazonicus (Savage & Peters) from mature nymphs to subimago stages. The sperm ultrastructure of Massartela brieni (Lestage), Farrodes carioca (Domínguez et al) and Miroculis mourei (Savage & Peters), as well as aspects of cell fragments observed in these species' subimagos deferent ducts were described. Sperm from the three species studied are aflagellated and immotile, while those from F. carioca and Ma. brieni are approximately spherical with a homogenous nucleus and acrosome. Sperm of F. carioca present two or three mitochondria located between the nucleus and the acrosome. In Ma. brieni, only one lateral mitochondria was found. Sperm from Mi. mourei are shaped as a number 'eight', with electron lucent spots inside the nucleus and two mitochondria above the acrosome. Large cell fragments containing degenerative vesicles and some sperm were observed in the deferent duct lumen of the three species. Testes of Mi. amazonicus are extremely reduced in the subimago stage, which suggests that these cell fragments originated from testes degeneration.

Morphometry of the midgut of Melipona quadrifasciata anthidioides (Lepeletier) (Hymenoptera: Apidae) during metamorphosis.

In Hymenoptera, midgut changes begin in the last instar. At this stage, the larval epithelial digestive cells degenerate, leaving only the basal membrane and the regenerative cells which will develop into a new epithelium during the pupal stage and in the adult. Epithelium renewal is followed by changes in volume and shape of the midgut. Morphometric analysis of digestive cells and total midgut volume of Melipona quadrifasciata anthidioides (Lepeletier) were conducted to verify whether cell volume increase are sufficient to account for the total midgut volume increase that occurs during metamorphosis. An increase in midgut volume was verified in spite of the scarcity of cell proliferation found during metamorphosis. At the end of metamorphosis, the increase in cell volume was not sufficient to explain the increase in volume of the midgut, indicating that an increase in the number of digestive cells is apparently necessary. Nevertheless, the mechanism by which regenerative cells reconstitute the epithelium during metamorphosis remains unknown.

Sperm morphology of the Prorops nasuta (Waterston, 1923) (Hymenoptera: Bethylidae).

In the present study, spermatozoa of the Prorops nasuta (Hymenoptera: Bethylidae) parasitoid were described morphologically. This is the first publication to describe a species belonging to the superfamily Chrysidoidea. Light and transmission electron microscopy were used. The spermatozoa of P. nasuta are linear, with a mean length of 665microm. The acrosome is composed of an acrosomal vesicle and a perforatorium. The nucleus measures approximately 17microm in length and is circular at its cross-section; however, its anterior extremity is oval. The chromatin is electron-dense and compact, although there are clear areas in the posterior peripheral regions. In the nucleus-flagellum transition region, the cross-section of the centriole adjunct is oval, with a pleated border and an E-PTA-positive peripheral region. The axoneme shows a 9+9+2 microtubule arrangement. The microtubules are E-PTA positive and, at the posterior extremity, the accessories are the last to terminate. The diameters and shapes of the two mitochondrial derivatives are almost identical. One begins beside the nuclear base and the other after the centriole adjunct. Posteriorly, they terminate together, immediately before the axoneme. Both have mitochondrial cristae and a region of paracrystalline material; however, the format and arrangement of this material differs from those of all other species previously studied. The paracrystalline material is more strongly E-PTA positive than the cristae region. Accessories bodies are electron-dense and located between the mitochondrial derivatives and the axoneme. In general, P. nasuta spermatozoa are similar to those of the majority of Hymenoptera; however, they have various exclusive characteristics that may be useful for studying the phylogeny and taxonomy of the superfamily Chrysidoidea and of Hymenoptera in general.

Seasonal changes in testicular and epididymal histology of the tropical lizard, Tropidurus itambere (Rodrigues, 1987), during its reproductive cycle.

The reproductive cycles of lizards, including Tropidurus species, have been widely studied. However, few studies describe in detail the ultrastructure and the epithelial changes in the epididymis. Using histology and trasmission electron microscopy, we show the seasonal changes in the testis and epididymis of the lizard Tropidurus itambere, during its annual reproductive cycle. The reproductive cycle of T. itambere was analysed from June 1988 to June 1989 and from June 2001 to June 2002. While the frequency of reproductive males in the population varied throughout the year, there were reproductive males in most months except for February through April. During this nonreproductive period, there is a reduction in the mean seminiferous tubule volume and few sperm were found in both the testis and the epididymis.

Seasonal changes in testicular and epididymal histology of the tropical lizard, Tropidurus itambere (Rodrigues, 1987), during its reproductive cycle / Mudanças sazonais na histologia do testículo e epidídimo do lagarto tropical Tropidurus itambere, durante seu ciclo reprodutivo

The reproductive cycles of lizards, including Tropidurus species, have been widely studied. However, few studies describe in detail the ultrastructure and the epithelial changes in the epididymis. Using histology and trasmission electron microscopy, we show the seasonal changes in the testis and epididymis of the lizard Tropidurus itambere, during its annual reproductive cycle. The reproductive cycle of T. itambere was analysed from June 1988 to June 1989 and from June 2001 to June 2002. While the frequency of reproductive males in the population varied throughout the year, there were reproductive males in most months except for February through April. During this nonreproductive period, there is a reduction in the mean seminiferous tubule volume and few sperm were found in both the testis and the epididymis.

O ciclo reprodutivo de alguns lagartos já foi extensamente estudado, incluindo algumas espécies do gênero Tropidurus. Entretanto, poucos estudos mostram os aspectos morfológicos do testículo e epidídimo, durante o ciclo reprodutivo anual, e não existe nenhuma descrição ultraestrutural das variações epiteliais no epidídimo. O presente estudo foi feito para mostrar, através da histologia e microscopia eletrônica de transmissão, detalhes das mudanças sazonais no testículo e epidídimo durante o ciclo reprodutivo anual. Acompanhamos o ciclo de Tropidurus itambere, na floresta atlântica brasileira, durante o período de junho de 2001 a junho de 2002. Os machos variam em sua atividade reprodutiva, mas foram considerados potencialmente reprodutivos na maioria dos meses, à exceção dos meses de fevereiro a abril. Durante este período, ocorreu uma redução no volume médio dos túbulos seminíferos e uma menor quantidade de espermatozóides no testículo e no epidídimo.

Sperm morphology of the neotropical harvestman Iporangaia pustulosa (Arachnida: Opiliones): comparative morphology and functional aspects.

We describe herein the sperm morphology of the harvestman Iporangaia pustulosa. Adult males were dissected, the reproductive tract was schematized and the seminal vesicle was processed by light, transmission and scanning electron microscopy. The male reproductive tract is composed of a tubular testis, two deferent ducts, a seminal vesicle, a propulsive organ and a penis, similar to that observed in other Opiliones. The spermatozoa from the seminal vesicle are oval, aflagellate and immotile, presenting a nucleus surrounding an invagination of the cytoplasm, as well as a complex acrosome and projections on the cell surface. In the testis, spermatozoa are devoid of projections. In the seminal vesicle, they gradually acquire the projections with tufts adhering to it. Consequently, spermatozoa in various distinct stages of projection development can be found in the seminal vesicle. We believe that these projections (1) could help transport sperm along the male and perhaps female reproductive tracts; (2) are used to anchor the spermatozoa inside the female spermatheca in order to avoid mechanical displacement by the genitalia of other males and (3) may play a role in oocyte recognition. We propose that the evolution of aflagellarity in Opiliones is related to the unique morphology of the female reproductive tract. Since eggs are fertilized on the tip of the ovipositor just prior to being laid, there is no advantage favoring sperm mobility. Additionally, female sperm receptacles are small and males that produced small spermatozoa would have a higher chance of fertilizing more eggs.

Use of blends of bioabsorbable poly(L-lactic acid)/poly(hydroxybutyrate- co-hydroxyvalerate) as surfaces for Vero cell culture.

Vero cells, a cell line established from the kidney of the African green monkey (Cercopithecus aethiops), were cultured in F-10 Ham medium supplemented with 10% fetal calf serum at 37 degrees C on membranes of poly(L-lactic acid) (PLLA), poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and their blends in different proportions (100/0, 60/40, 50/50, 40/60, and 0/100). The present study evaluated morphology of cells grown on different polymeric substrates after 24 h of culture by scanning electron microscopy. Cell adhesion was also analyzed after 2 h of inoculation. For cell growth evaluation, the cells were maintained in culture for 48, 120, 240, and 360 h. For cytochemical study, the cells were cultured for 120 or 240 h, fixed, processed for histological analysis, and stained with Toluidine blue, pH 4.0, and Xylidine ponceau, pH 2.5. Our results showed that cell adhesion was better when 60/40 and 50/50 blends were used although cells were able to grow and proliferate on all blends tested. When using PLLA/PHBV (50/50) slightly flattened cells were observed on porous and smooth areas. PLLA/PHBV (40/60) blends presented flattened cells on smooth areas. PLLA/PHBV (0/100), which presented no pores, also supported spreading cells interconnected by thin filaments. Histological sections showed that cells grew as a confluent monolayer on different substrates. Cytochemical analysis showed basophilic cells, indicating a large amount of RNA and proteins. Hence, we detected changes in cell morphology induced by alterations in blend proportions. This suggests that the cells changed their differentiation pattern when on various PLLA/PHBV blend surfaces.

Use of blends of bioabsorbable poly(L-lactic acid)/poly(hydroxybutyrate- co-hydroxyvalerate) as surfaces for Vero cell culture

Vero cells, a cell line established from the kidney of the African green monkey (Cercopithecus aethiops), were cultured in F-10 Ham medium supplemented with 10 percent fetal calf serum at 37°C on membranes of poly(L-lactic acid) (PLLA), poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and their blends in different proportions (100/0, 60/40, 50/50, 40/60, and 0/100). The present study evaluated morphology of cells grown on different polymeric substrates after 24 h of culture by scanning electron microscopy. Cell adhesion was also analyzed after 2 h of inoculation. For cell growth evaluation, the cells were maintained in culture for 48, 120, 240, and 360 h. For cytochemical study, the cells were cultured for 120 or 240 h, fixed, processed for histological analysis, and stained with Toluidine blue, pH 4.0, and Xylidine ponceau, pH 2.5. Our results showed that cell adhesion was better when 60/40 and 50/50 blends were used although cells were able to grow and proliferate on all blends tested. When using PLLA/PHBV (50/50) slightly flattened cells were observed on porous and smooth areas. PLLA/PHBV (40/60) blends presented flattened cells on smooth areas. PLLA/PHBV (0/100), which presented no pores, also supported spreading cells interconnected by thin filaments. Histological sections showed that cells grew as a confluent monolayer on different substrates. Cytochemical analysis showed basophilic cells, indicating a large amount of RNA and proteins. Hence, we detected changes in cell morphology induced by alterations in blend proportions. This suggests that the cells changed their differentiation pattern when on various PLLA/PHBV blend surfaces.

Immunocytochemical localization of tubulins in spermatids and spermatozoa of Euptoieta hegesia (Lepidoptera: Nymphalidae).

A comparative analysis of the distribution of tubulin types in apyrene and eupyrene sperm of Euptoieta hegesia butterflies was carried out, also verifying the presence of tubulin in lacinate appendages of the eupyrene sperm. Ultrathin sections of LR White embedded spermatids and spermatozoa were labeled for alpha, beta, gamma, alpha-acetylated and alpha-tyrosinated tubulins. Apyrene and eupyrene spermatids show the same antibody recognition pattern for tubulins. All tubulin types were detected in axonemal microtubules. Alpha and gamma tubulins were also detected on the cytoplasmic microtubules. However, for beta and tyrosinated tubulins only scattered labeling was detected on cytoplasmic microtubules and acetylated tubulin was not detected. In apyrene and eupyrene spermatozoa only the axoneme labeling was analyzed since cytoplasmic microtubules no longer exist in these cells. Alpha, beta and tyrosinated tubulins were easily detected on the apyrene and eupyrene axoneme; gamma tubulin was strongly marked on eupyrene axonemes but was scattered on the apyrene ones. Acetylated tubulin appeared with scattered labeling on the axoneme of both sperm types. Our results demonstrate significant differences in tubulin distribution in apyrene and eupyrene axonemal and cytoplasmic microtubules. Extracellular structures, especially the lacinate appendages, were not labeled by antibodies for any tubulin.

Effects of anoxia on root ultrastructure of four neotropical trees.

The root ultrastructure of seedlings grown in anaerobic conditions was investigated in four neotropical species: Sesbania virgata, Erythrina speciosa, Sebastiania commersoniana (all present in waterlogged or flooded areas), and Schizolobium parahyba (that occupies mainly dry areas). Anaerobiosis induced an increase in the size of mitochondria, dilatation of cristae and of the endoplasmic reticulum, and fragmentation or concentric arrangement of reticulum saccules. The ultrastructural alterations were reversible only for S. virgata and E. speciosa. The seedlings of S. parahyba and S. commersoniana were more sensitive to oxygen deprivation and presented extensive cell disruption. The results are discussed in terms of energy supply.