Studies on charge transfer of enalapril maleate: from solid-state to molecular dynamics.

INTRODUCTION: Enalapril maleate is an antihypertensive ethyl ester pro-drug with two crystalline forms. A network of hydrogen bonds in both polymorphs plays an important role on solid-state stability, charge transfer process and degradation reactions (when exposed to high humidity, temperature and/or pH changes). COMPUTATIONAL PROCEDURES: Supramolecular arrangement was proposed by Hirshfeld surface using the CrystalExplorer17 software and quantum theory of atoms in molecules. The electronic structure properties were calculated using the functional hybrid M06-2X with 6-311++G** base function employing diffuse and polarization functions to improve the description of hydrogen atoms on intermolecular interactions. Also, the H+ charge transfer between enalapril and maleate molecules was performed using Car-Parrinello molecular dynamics with the Verlet algorithm. In both simulations, the temperature of the ionic system was maintained around 300 K using the Nosé-Hoover thermostat and the electronic system evolved without the use of the thermostat. RESULTS: This work evaluates the effect of maleate on the structural stability of enalapril maleate solid state. The electronic structural analysis points out a partially covalent character for N1-H∙∙∙O7 interaction; and the molecular dynamic showed a decentralized hydrogen on maleate driving a decomposition by charge transfer process while a centered hydrogen driving the stabilization. The charge transfer process and the mobility of the proton (H+) between enalapril and maleate molecules was demonstrated using supramolecular modeling analyses and molecular dynamics calculations.

A new species of the Scinax catharinae species group (Anura: Hylidae) from the Atlantic rainforest of northeastern Minas Gerais, southeastern Brazil.

We describe a new species of the Scinax catharinae group from one of the last remaining fragments of Atlantic Forest in northeastern Minas Gerais. The new species is distinguished from most species of the group mainly by having the mucronate snout in dorsal view, the unique light-yellow coloration on the gular surface in life contrasting with its overall color pattern, and the m. depressor mandibulae with an origin on the dorsal fascia at the level of the m. dorsalis scapulae. These two latter features are novelties in the diagnosis of Scinax species. The new species is currently known only from municipality of Santa Maria do Salto, state of Minas Gerais, Brazil. This locality is a remaining fragment of Atlantic Forest and has a peculiar geomorphic feature that has been poorly sampled so far.

The tadpole of Physalaemus orophilus from the Atlantic rainforest of southeastern Brazil (Amphibia, Anura, Leptodactylidae).

The neotropical genus Physalaemus Fitzinger is currently composed by 48 species (Cardozo Pereyra 2018). Recently, a phylogenetic analysis aiming to investigate the internal relationships of the genus recovered two major clades, the Physalaemus cuvieri and P. signifer clades (Lourenço et al. 2015). The following species groups were retrieved in the first clade: P. biligonigerus, P. cuvieri, P. gracilis, P. henselii, and P. olfersii groups (Lourenço et al. 2015). This proposal redefined the P. olfersii group including P. olfersii (Lichtenstein Martens), P. soaresi Izecksohn, P. maximus Feio, Pombal, Caramaschi, P. feioi Cassini, Cruz, Caramaschi and P. lateristriga (Steindachner). The authors also allocated tentatively P. orophilus Cassini, Cruz, Caramaschi, and P. insperatus Cruz, Cassini, Caramaschi in the P. olfersii group due to their morphological similarity with the other species (Cruz et al. 2008; Cassini et al. 2010). Otherwise, Physalaemus aguirrei Bokermann was not recovered nested within this group, contradicting what was suggested in a previous phenetic analysis (Nascimento et al. 2005). Members of the P. olfersii group inhabit the Atlantic rainforest and most of them have a similar advertisement calls with pulsed notes, without frequency modulation and harmonic structure (Giaretta et al. 2009; Cassini et al. 2010; Lourenço et al. 2015). Regarding their larval stage, only P. soaresi, P. maximus, and P. olfersii have their tadpoles described (Weber et al. 2005; Baêta et al. 2007; Giaretta et al. 2009). Physalaemus orophilus occurs in montane Atlantic Forest sites at the eastern slope of the Espinhaço Range in the State of Minas Gerais, southeastern Brazil (Cassini et al. 2010). Herein, we describe the tadpole of P. orophilus from Quadrilátero Ferrífero mountain region, southern limit of the Espinhaço Range and compared it to the known tadpoles of the P. olfersii group.

Osteology of Physalaemus nattereri (Anura: Leptodactylidae) with comments on intraspecific variation.

The cranium, postcranium, and osteological variation of Physalaemus nattereri (Steindachner) are described. The main sources of variation involve the degree of mineralization of the nasal capsule and the lengths of dermal skull bones (e.g., vomer, sphenethmoid, and neopalatine). Osteologically, P. nattereri differs from its congeners by the anterior placement of the jaw articulation (which lies anterior to the intersection between the alae and cultriform process of parasphenoid), and by the separation of the frontoparietals from the anterior margins of exoccipitals. Descriptions of the nasal capsule, the auditory apparatus, and the iliosacral articulation are presented for the first time for this species. One putative morphological synapomorphy is presented for the P. signifer Clade.

A new species of the Scinax catharinae group (Anura: Hylidae) from southeastern Brazil.

We describe a new species of the Scinax catharinae group from coastal forest in the State of São Paulo, Brazil. We also describe the karyotype and the advertisement call of the new species, which is composed by a multipulsed note. The chromosome morphology of the new species shows the nucleolar organizer region (NOR) on Pair 6, a putative synapomorphy previously inferred for the S. catharinae clade. We report, for the first time, the presence of mental glands in a species of the genus Scinax. Additionally, we provide a brief discussion about the occurrence of glandular tissue on the mental and inguinal regions of the species of the S. catharinae clade.

Morphological variation, advertisement call, and tadpoles of Bokermannohyla nanuzae (Bokermann, 1973), and taxonomic status of B. feioi (Napoli & Caramaschi, 2004) (Anura, Hylidae, Cophomantini).

Bokermannohyla nanuzae (Bokermann & Sazima 1973) and B. feioi (Napoli & Caramaschi 2004) belong to the B. cir-cumdata species group. The type locality of the former is Serra do Cipó, Espinhaço mountain range, and of the latter is Parque Estadual do Ibitipoca, Mantiqueira mountain range, both in Minas Gerais State, Brazil. Differences on dorsal draw-ing pattern of adults, oral disc morphology of tadpoles, and temporal properties of calls were proposed to distinguish these two species. However, several specimens found between the two type localities remain unidentified because diagnostic characters and states occur in all of these populations. Thus, in order to assess these characters variations, we performed an analysis of the morphology and morphometry of adults, vocalization, and morphology of tadpoles. Specimens were divided into three operational taxonomic units (OTUs): B. nanuzae (Serra do Cipó and northwards, Espinhaço mountain range), B. cf. nanuzae (Quadrilátero Ferrífero, Espinhaço mountain range, south of Serra do Cipó), and B. feioi (Serra do Ibitipoca, Mantiqueira mountain range). Drawing patterns of the dorsum and limbs show clinal variation and the three units are morphometrically very similar. Temporal and spectral properties of calls overlap in these three units. The diagnostic differences originally proposed for tadpoles are intrapopulational variations and occur in specimens from all of the locations analyzed. We found that these three units are morphologically indistinguishable. Therefore, we designate Bok-ermannohyla feioi (Napoli & Caramaschi 2004) as a junior synonym of Bokermannohyla nanuzae (Bokermann & Sazima 1973), extending its geographical distribution to the Mantiqueira mountain range.

Supplementation with Pfaffia glomerata (Sprengel) Pedersen does not affect androgenic-anabolic parameters in male rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Paffia spp (Amaranthacea) has a widespread use of in Brazil as a possible hormonal supplement and a substitute of Panax ginseng, although information on its reproductive effects is missing. AIM OF THE STUDY: To evaluated possible anabolic-androgenic or anti-androgenic effects of Pfaffia glomerata (PG) extract using intact eight-months-old male rats and pre-pubertal castrated rats. MATERIALS AND METHODS: Three different dose levels of PG (8.5, 30 and 85 mg/kg/day) were administered to eight-months-old rats for 28 days or to castrated males for 7 days (Hershberger assay). In the experiment with intact animals, 24h fecal samples were collected for quantification of fecal metabolites of androgens throughout treatment. At the end of the treatment period, animals were euthanized for evaluation of serum testosterone, reproductive organ weights, number of spermatids per testis, diameter of seminiferous tubules and cross-sectional area of soleus muscle fibers. In the Hershberber assay, androgenic or anti-androgenic effects were evaluated by the weights of androgen-dependent tissues: ventral prostate, seminal vesicle, glans penis and levator ani muscle/bulbocavernosus muscle. RESULTS: No effects were observed in the concentrations of fecal metabolites of androgens monitored during the treatment of intact eight-months-old rats. Moreover, at the end of treatment, no changes were seen in any of the investigated parameters. In the Hershberger assay, the PG extract did not induce androgenic or anti-androgenic effects at the dose levels tested. Significant effects were only observed in animals treated with testosterone and testosterone plus flutamide, which were used as positive controls for androgenicity and anti-androgenicity, respectively. CONCLUSIONS: At the dose levels tested, PG extract does not induce anabolic-androgenic or anti-androgenic effects in rats.

A new species of the Scinax catharinae Group (Anura: Hylidae) from Northeastern Brazil.

We describe a new species of the Scinax catharinae Group from Municipality of Porto Seguro, State of Bahia northeastern Brazil. The new species is mainly characterized by its small size, nuptial pad dark colored, and compound pectoral fold. Additionally, we describe the structure of its nuptial pad and compare it with that of S. agilis. We also briefly discuss its phylogenetic relationships within Scinax. 

Effects of ß-glucan polysaccharide revealed by the dominant lethal assay and micronucleus assays, and reproductive performance of male mice exposed to cyclophosphamide.

ß-glucan is a well-known polysaccharide for its chemopreventive effect. This study aimed to evaluate the chemopreventive ability of ß-glucan in somatic and germ cells through the dominant lethal and micronucleus assays, and its influence on the reproductive performance of male mice exposed to cyclophosphamide. The results indicate that ß-glucan is capable of preventing changes in DNA in both germ cells and somatic ones. Changes in germ cells were evaluated by the dominant lethal assay and showed damage reduction percentages of 46.46% and 43.79% for the doses of 100 and 150 mg/kg. For the somatic changes, evaluated by micronucleus assay in peripheral blood cells in the first week of treatment, damage reduction percentages from 80.63-116.32% were found. In the fifth and sixth weeks, the percentage ranged from 10.20-52.54% and -0.95-62.35%, respectively. Besides the chemopreventive efficiency it appears that the ß-glucan, when combined with cyclophosphamide, is able to improve the reproductive performance of males verified by the significant reduction in rates of post-implantation losses and reabsorption in the mating of nulliparous females with males treated with cyclophosphamide.

Effects of Angiotensin-Converting Enzyme Inhibitor Derived from Tropaeolum majus L. in Rat Preimplantation Embryos: Evidence for the Dehydroepiandrosterone and Estradiol Role.

Although several studies have shown the inhibitory effects of Tropaeolum majus extracts (HETM) on angiotensin-converting enzyme (ACE) activity, no studies have been carried out during the beginning of pregnancy, when humoral and hormonal imbalance may affect zygote and early embryo transport. This study investigates whether HETM can affect embryonic development when administered during the one-cell-blastocyst period. Pregnant Wistar rats received orally the HETM (3, 30, and 300 mg/kg/day) from the 1st to the 7th gestational day. Rats were killed on the 8th day of pregnancy and the following parameters were evaluated: clinical symptoms of toxicity (including organ weights), number of corpora lutea, implants per group, preimplantation losses ratio, and the serum levels of dehydroepiandrosterone (DHEA), estradiol, and progesterone. No clinical symptoms of maternal toxicity were evidenced. On the 8th day of pregnancy, the levels of DHEA and estradiol were increased and significant preimplantation losses were observed at all doses used. The present study reveals that the HETM can raise levels of DHEA and estradiol and induce difficulty in the embryo implantation in the early stages of pregnancy. The data contributes significantly to the safety aspects of using this natural product when trying to get pregnant or during pregnancy.

Testicular effects following in utero exposure to the antivirals acyclovir and ganciclovir in rats.

In utero exposure to the antivirals acyclovir and ganciclovir has been reported to induce gross structural defects in rat offspring. The present study investigated the effects of maternal antiviral treatment on gestation day 10 on reproductive and nonreproductive organs in male rat offspring with a particular focus on the testes. Vehicle and two doses of acyclovir and ganciclovir, 75 and 300 mg/kg, were administered to rat dams. The total doses were fractioned into three subcutaneous applications (3 × distilled water, 3 × 25 mg/kg, and 3 × 100 mg/kg) that were administered on gestation day 10 at 8:00 a.m., 1:00 p.m., and 6:00 p.m. The antiviral concentrations were measured in the serum of the dams 1 h after the last administration. Exposure to 300 mg/kg ganciclovir induced germ cell deficiency in both fetal and adult testes, an effect that was not seen in any other treatment group. Adult rats exposed in utero to this high ganciclovir dose exhibited Sertoli cell-only tubules intermingled with seminiferous tubules that displayed a normal size and normal cell counts, alterations that resemble focal Sertoli cell-only syndrome in humans. The serum concentrations of ganciclovir were markedly higher than those of acyclovir, particularly at the high dose tested. However, although 300 mg/kg acyclovir did not induce germ cell deficiency, other specific effects were seen in exposed animals, including incomplete eye opening and reduced thymus weight.

In vivo evaluation of the antimutagenic and antigenotoxic effects of ß-glucan extracted from Saccharomyces cerevisiae in acute treatment with multiple doses.

Ample evidence suggests that cancer is triggered by mutagenic damage and diets or supplements capable of reducing such incidences can be related to the prevention of neoplasy development or to an improvement in life quality of patients who undergo chemotherapy. This research aimed to evaluate the antimutagenic and antigenotoxic activity of ß-glucan. We set up 8 experimental groups: control (Group 1), cyclophosphamide (Group 2), Groups 3-5 to assess the effect of ß-glucan administration, and Groups 6-8 to evaluate the association between cyclophosphamide and ß-glucan. The intraperitonial concentrations of ß-glucan used were 100, 150 and 200 mg/kg. Micronucleus and comet assays showed that within the first week of treatment ß-glucan presented a damage reduction rate between 100-62.04% and 94.34-59.52% for mutagenic and genotoxic damages, respectively. This activity decreased as the treatment was extended. During the sixth week of treatment antimutagenicity rates were reduced to 59.51-39.83% and antigenotoxicity was not effective. This leads to the conclusion that the efficacy of ß-glucan in preventing DNA damage is limited when treatment is extended, and that its use as a chemotherapeutic adjuvant need to be better clarified.

A new species of the Scinax catharinae group (Anura, Hylidae) from Serra da Canastra, southwestern state of Minas Gerais, Brazil.

We describe Scinax pombali sp. n. a new species of treefrog of the Scinax catharinae group from Serra da Canastra, municipality of Capitólio (20°36'03''S, 46°17'34.9''W, 987 m a.s.l.), located in the Cerrado domains of the State of Minas Gerais, Southeastern Brazil. The new species is characterized by its small size, blotches and color pattern on dorsal surface and hidden regions of flanks and thighs, canthus rostralis lightly concave and well marked, absent nuptial pad, and lack of externally differentiated inguinal gland. Additionally, we describe the tadpole of this new species, which is characterized by the large-sized oral disc and presence of a large number of marginal papillae (two to three rows on its dorsal portion and some rows in unorganized arrangement on its lateroventral portion).

Study on the developmental toxicity of combined artesunate and mefloquine antimalarial drugs on rats.

Antimalarial drug combinations containing artemisinins (ACTs) have become first choice therapies for Plasmodium falciparum malaria. Data on safety of ACTs in pregnancy are limited and no previous study has been conducted on the developmental toxicity of artesunate-mefloquine combinations on the first trimester of gestation. To evaluate the developmental toxicity of an artesunate/mefloquine combination, pregnant rats were treated orally with artesunate (15 and 40 mg/kg bwt/day), mefloquine (30 and 80 mg/kg bwt/day) and artesunate/mefloquine (15/30 and 40/80 mg/kg bwt/day) on gestation days 9-11. Dams were C-sectioned on day 20, and their uteri and fetuses removed and examined for soft tissue and skeleton abnormalities. Artesunate increased embryolethality and the incidence of limb long bone malformations on the absence of overt maternal toxicity. Mefloquine (80 mg/kg bwt/day) was maternally toxic and enhanced fetal variations. Combination of artesunate and mefloquine did not enhance their toxicity compared to the toxicity observed after its separate administration. Embryotoxicity of artesunate was apparently attenuated when it is co-administered with mefloquine.

Ocupação da coluna da água por machos de Trichogaster leeri (Bleeker, 1852) (Perciformes, Osphronemidae) em aquário / Water column occupation by males of Trichogaster leeri (Bleeker, 1852) (Perciformes, Osphronemidae) in aquarium

Trichogaster leeri é considerado um peixe pacífico e muito utilizado em aquarismo pela beleza de suas cores. O presente estudo teve como objetivos avaliar a ocupação da coluna da água por machos adultos quando mantidos com outros machos da mesma espécie. Foram utilizados três aquários com as dimensões de 35 cm de comprimento, 22 cm de altura e 16 cm de largura, com substrato de cascalho e vegetação artificial. O vidro frontal foi dividido em 12 quadrantes, ordenados da esquerda para a direita e de cima para baixo. Três exemplares de aproximadamente 8 cm de comprimento total foram utilizados para observação das freqüências de ocorrência nos quadrantes, nas seguintes situações: isolados, agrupados em duplas e os três juntos. Os dados foram analisados pelo teste χ 2, ao nível de significância de 5%. Foi observado que, quando associados em um mesmo aquário, os indivíduos ocupam uma determinada posição na coluna da água. Isso significa que, apesar do comportamento aparentemente pacífico, os machos se evitam ocupando distintas posições espaciais. .

Trichogaster leeri is considered a peaceful fish and widely used in aquariums for the beauty of its colors. This study aimed to evaluate the occupation of water column by adult males when kept with other males of the same species. Three 35 cm-long, 22 cm-high and 16 cm-wide aquariums were used, with grave as substrate and artificial vegetation. The front glass was divided in 12 quarters, arranged from left to right and from top to bottom. Three 8 cm-long specimens were used to observe the frequency of occurrence in the quarters in the following situations: isolated, grouped in pairs and all of them together. The data were analyzed by χ² test, at significance level of 5%. It was observed that, when the individuals are together in the same aquarium, they occupy a position in the water column. It means that, in spite of the apparently pacific behavior, the males avoid each other occupying distinct spatial positions..

Evaluation of subchronic toxicity of the hydroethanolic extract of Tropaeolum majus in Wistar rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The use of medicinal plants for the treatment of diseases usually comes from the belief that they present low toxicity due their natural origin. However, it is necessary a toxicological and pharmacological evaluation for these plants. Tropaeolum majus is a medicinal plant used in popular medicine to treat several diseases, including cardiovascular disorders, urinary tract infections and asthma. Even though several studies proved its therapeutic effects, there are few toxicological studies with this species. AIM OF THE STUDY: The present study was carried out to evaluate the subchronic toxicity of the hydroethanolic extract obtained from leaves of T. majus (HETM) in Wistar rats. MATERIAL AND METHODS: Male and female Wistar rats received three doses of HETM (75, 375 and 750 mg/kg) for 28 days. After the treatments biochemical, hematological and histopathological parameters were analyzed. RESULTS: No significant alterations in the animal's body weight gain, relative organs weight, serum biochemical analysis, hematological or histopathological analyses of liver, kidneys and spleen were observed. CONCLUSIONS: These results demonstrate the absence of subchronic toxicity due to oral treatment with HETM for 28 days in Wistar rats. However, other toxicological studies are necessary to evaluate the total safety of this plant.

Screening for in vivo (anti)estrogenic and (anti)androgenic activities of Tropaeolum majus L. and its effect on uterine contractility.

ETHNOPHARMACOLOGICAL RELEVANCE: Tropaeolum majus L. (Tropaeolaceae) is a medicinal herb popularly used in Brazil for treatment of inflammatory and cardiovascular diseases. Despite some published data on its efficacy, there are still few toxicological data describing the safety of this plant. The aim of this study was to evaluate the (anti)estrogenic and (anti)androgenic activity of the hydroethanolic extract obtained from Tropaeolum majus L. (HETM), as well as its possible effects on uterine contractility. MATERIALS AND METHODS: Three experimental protocols were performed, (a) uterotrophic assay, (b) Hershberger assay and (c) an ex vivo test to investigate the effects of maternal administration of HETM on uterine contractility at the end of pregnancy. In all protocols three doses of the HETM were administered to Wistar rats: 3, 30 and 300mg/kg. RESULTS: In vivo tests for detection of (anti)androgenic and (anti)estrogenic activities did not show any significant alterations. Similarly, no alterations were observed on uterine contractility induced by oxytocin and arachidonic acid. CONCLUSIONS: HETM was unable to produce (anti)estrogenic or (anti)androgenic activities in the short-term in vivo screening assays performed. In addition, there was no evidence that HETM can affect uterine contractility following gestational exposure of rats.

Effects of the polysaccharide beta-glucan on clastogenicity and teratogenicity caused by acute exposure to cyclophosphamide in mice.

Alterations that could lead to the cancer development may also be related to an adverse development of offspring in experimental animals. Some functional foods, which contain the polysaccharide beta-glucan, have been described as being effective in the prevention of clastogenic damage. Based on that finding, the aim of the present study was to determine the efficacy of this sugar polymer in the mutagenic and teratogenic damage control. Two sets of females, pregnant and non-pregnant, were evaluated. The results indicated that beta-glucan was effective in preventing clastogenic damage in pregnant as well as non-pregnant females. In addition, pregnant females were more susceptible to mutagenic damage. However, teratogenic effects were not prevented effectively, although there was a trend toward a reduction in level of malformations. Despite beta-glucan did not prevent malformations, it increased fetal viability and reduced number of post-implantation losses and resorption, thereby enhancing reproductive performance in females.