RESUMO
Urban environments present less environmental heterogeneity in relation to the natural ones, affecting the biodiversity of bats and the ecological processes in which they participate. In this way, we will identify how urbanization influences the structure of bat communities in the municipality of Goiânia, Goiás, Brazil. We compared species composition, guilds and bat richness in a gradient that crossed urban, semi-urban and natural areas in the municipality of Goiânia, contained in the Cerrado biome. We captured a total of 775 bats of 16 species distributed in three families. Urban areas had a higher species abundance, while semi-urban areas had a higher species richness. The three types of environments have different compositions, the urban one being more homogeneous, the fauna in these areas is composed of generalist species, which benefit from this process. The diversity present in semi-urban areas is a consequence of the intersection between urban and natural fauna, which is why urban expansion needs to occur in a planned manner to minimize the impacts of this process and ensure the maintenance of biodiversity.
Assuntos
Quirópteros , Humanos , Animais , Urbanização , Brasil , Pradaria , Ecossistema , BiodiversidadeRESUMO
The present study aims to establish the morphological, morphometric, and immunostaining patterns of the steroidogenic enzymes 17ß-HSD and 5α-reductase and androgen receptors (AR) during the prenatal development of the male gonad and epididymis of Cavia porcellus. Fetuses at 22, 25, 30, 40, 45, 50, and 60 days of gestation (DG) were used. Specimens were dissected and subjected to macroscopic, histological, histomorphometric, and immunohistochemical analyses. Genital and scrotal protrusions were identified in 22 DG embryos. Gonocytes were identified at 25 DG and the formation of primary testicular cords was observed at 30 DG. Through anatomical evaluation, we observed differentiation of the epididymis into the head, body, and tail at 45 DG. During development, there is a progressive decrease in the diameters of the testicular cords and epididymal ducts. 17ß-HSD enzyme immunostaining was observed in Leydig cells at all ages, while 5α-reductase was observed in Leydig cell cytoplasm and gonocytes at 40, 50, and 60 DG. AR shows gonocyte labeling at 30 DG. Thus, from the second trimester of pregnancy, it is possible to observe patterns of anatomical development, such as genital and scrotal prominence (22 DG), the appearance of gonocytes in the testicular cords at 25 DG, and the beginning of the organization of primary testicular cords at 30 DG, suggesting sexual differentiation. The 17ß-HSD, 5α-reductase, and ARs play an essential role in sexual development and differentiation, presenting immunostaining at different reproductive process times.
Assuntos
Epididimo , Testículo , Gravidez , Feminino , Cobaias , Masculino , Animais , Células Intersticiais do Testículo , Oxirredutases , Receptores AndrogênicosRESUMO
This study aims to define the best method (slow freezing or vitrification) and fragment size (1, 5, or 9 mm³) for prepubertal goat testis cryopreservation, as well as to evaluate testicular morphological integrity after cryopreservation and in vitro culture (IVC). Initially (experiment I), 1, 5, or 9 mm³ testis fragments were cryopreserved by slow freezing using a Mr. Frosty container with 20% Dimethylsulfoxide (DMSO) or vitrified using the Ovarian Tissue Cryosystem (OTC) device, (Equilibration solution - ES: 10% DMSO and 10% ethylene glycol - EG; Vitrification solution - VS: 20% DMSO and 20% EG) and then subjected to morphological analysis, type I and III collagen quantification and gene expression (Oct4, C-kit, Bax, and Bcl-2). Subsequently, (experiment II), fresh or cryopreserved by slow freezing testis fragments were cultured in vitro and submitted to morphological analysis by scanning electron microscopy. The data from the experiment I revealed fewer morphological alterations in 1 and 5 mm³ fragments after vitrification and slow freezing, respectively. The percentage of type I collagen fibers in 5 and 9 mm³ frozen was higher than in fresh or vitrified fragments. For type III collagen, fresh or frozen fragments of 1 and 5 mm3 showed a higher percentage than fragments of 9 mm3. Gene expression for Oct4 and C-kit after slow freezing or vitrification in the 5 mm3 fragments was lower than that observed in the fresh fragments. The Bax:Bcl-2 ratio in the 1 and 9 mm³ fragments was lower than in the 5 mm³ fragments for fresh fragments or after freezing. In experiment II, fragments cultured in vitro, previously frozen or not, showed more morphological alterations than fresh or frozen fragments. We concluded that slow freezing of 5 mm³ fragments was the best protocol for cryopreserving prepubertal goat testis and although the results of IVC are encouraging, it still needs improvement to restore testicular function after cryopreservation.
Assuntos
Dimetil Sulfóxido , Cabras , Animais , Masculino , Proteína X Associada a bcl-2 , Criopreservação/veterinária , Proteínas Proto-Oncogênicas c-bcl-2 , Proteínas Proto-Oncogênicas c-kitRESUMO
Cervical cancer is a serious public health problem in Brazil, especially in Manaus (Amazonas), the city with the highest incidence rate of cervical cancer in the country. Persistent infection with oncogenic human papillomavirus (HPV) genotypes is the cause of disease development. The aim of this study was to investigate the prevalence of oncogenic genotypes in women at high risk for cervical precancer examined in two policlinics in Manaus. One hundred and two patients who underwent colposcopy took part in the research. The DNA samples obtained from the cervical epithelium were analyzed by PCR with type-specific primers for the detection of eight oncogenic genotypes, which were chosen based on previous studies. The presence of HPV virus was detected in all samples. The most prevalent oncogenic genotypes were 18 (47.1%) and 16 (45.1%). Interestingly, HPV 18 was considered uncommon in this region. In addition to these, genotypes 31 (19.6%), 58 (19.6%), 33 (18.6%), and 45 (15.7%) also had a relatively high frequency in this population. Fifty-six women (54.9%) had multiple infections with up to five oncogenic types. Also, the presence of genotypes other than 16 and 18 was observed in most samples (57.8%), which also deserves attention since they are not covered by currently available vaccines against HPV in Brazil. The high prevalence and multiple infections with several oncogenic HPV genotypes in association with precursor lesions for cervical cancer highlighted the need to improve strategies to prevent this disease in Amazonas.
Assuntos
Infecções por Papillomavirus , Neoplasias do Colo do Útero , Humanos , Feminino , Neoplasias do Colo do Útero/patologia , Papillomavirus Humano 18/genética , Infecções por Papillomavirus/complicações , Prevalência , Brasil/epidemiologia , Papillomaviridae/genética , DNA Viral/genética , DNA Viral/análise , GenótipoRESUMO
Cervical cancer is a serious public health problem in Brazil, especially in Manaus (Amazonas), the city with the highest incidence rate of cervical cancer in the country. Persistent infection with oncogenic human papillomavirus (HPV) genotypes is the cause of disease development. The aim of this study was to investigate the prevalence of oncogenic genotypes in women at high risk for cervical precancer examined in two policlinics in Manaus. One hundred and two patients who underwent colposcopy took part in the research. The DNA samples obtained from the cervical epithelium were analyzed by PCR with type-specific primers for the detection of eight oncogenic genotypes, which were chosen based on previous studies. The presence of HPV virus was detected in all samples. The most prevalent oncogenic genotypes were 18 (47.1%) and 16 (45.1%). Interestingly, HPV 18 was considered uncommon in this region. In addition to these, genotypes 31 (19.6%), 58 (19.6%), 33 (18.6%), and 45 (15.7%) also had a relatively high frequency in this population. Fifty-six women (54.9%) had multiple infections with up to five oncogenic types. Also, the presence of genotypes other than 16 and 18 was observed in most samples (57.8%), which also deserves attention since they are not covered by currently available vaccines against HPV in Brazil. The high prevalence and multiple infections with several oncogenic HPV genotypes in association with precursor lesions for cervical cancer highlighted the need to improve strategies to prevent this disease in Amazonas.
RESUMO
The aim of this study was to evaluate the effect of 1 µmol/l zearalenone (ZEN) and 1 µmol/l matairesinol (MAT), alone or in combination, on the morphology of in vitro-cultured ovarian preantral follicles. Ovaries from four adult sheep were collected at a local slaughterhouse and fragmented, and the ovarian pieces were submitted to in vitro culture for 3 days in the presence or absence of the test compounds. The morphology of primordial and primary follicles was impaired by ZEN. The plant lignan MAT alone did not maintain the morphology of the ovarian follicles; its combination with ZEN counteracted the negative effects observed when follicles were cultured in the presence of the mycotoxin alone. However, MAT was not able to promote the in vitro development of the ovarian follicles.
Assuntos
Lignanas , Zearalenona , Animais , Feminino , Furanos , Lignanas/farmacologia , Folículo Ovariano , Ovário , Ovinos , Zearalenona/toxicidadeRESUMO
Ovine ovarian fragments (3 × 3 × 1 mm) were fixed in neutral buffered formalin (NBF), Carnoy's solution (CAR), Davidson's solution (DAV), or paraformaldehyde (PFA) for 12 h or 24 h. After this fixation time, each fragment was prepared for histological analysis. Although fixative and fixation period did not affect follicular and stromal cells density, the percentages of morphologically normal primordial and primary follicles was affected by the fixative type and period of fixation. Paraformaldehyde was not indicated as a fixative for ovarian fragments. Formalin was a suitable fixative only when the period of fixation was 12 h, while Carnoy was efficient after a fixation period of 24 h. In conclusion, the most indicated fixative for the morphological evaluation of ovarian preantral follicles was DAV, regardless of the fixation period, that is 12 or 24 h.
Assuntos
Folículo Ovariano , Ovário , Animais , Feminino , Fixadores/farmacologia , Ovinos , Fixação de TecidosRESUMO
The aim of this study was to evaluate the effect of 1 µmol/L zearalenone (ZEN) and 1 µmol/L enterolactone (ENL), alone or in combination, on the survival and morphology of in vitro cultured ovarian preantral follicles. Ovaries from 10 sheep were collected at a local abattoir and fragmented, and the ovarian pieces were submitted to in vitro culture for 3 days in the presence or absence of the test compounds. The morphology of primordial and primary follicles was impaired by ZEN, whereas that of cultured secondary follicles was improved by ENL. However, the combination of ENL with ZEN impaired the quality of primary and secondary follicles. Both ZEN and ENL induced apoptosis, but only ZEN was responsible for oocyte autophagy. None of these xenoestrogens affected endoplasmic reticulum stress as observed by the unaltered expression of ERP29. Differently from ZEN, ENL increased the expression of the efflux transporter ABCG2. In conclusion, although ENL can counteract the negative effects of ZEN on primordial and primary follicles, this positive effect is not similar to that observed in ovarian tissue cultures in the presence of ENL alone.
Assuntos
Zearalenona , 4-Butirolactona/análogos & derivados , Animais , Feminino , Lignanas , Oócitos , Folículo Ovariano , Ovário , Ovinos , Zearalenona/toxicidadeRESUMO
This study assessed the effect of total dry matter intake (DMI), plant life form and strata harvested on the gastrointestinal nematode (GIN) worm burdens of tracer kids and lambs browsing heterogeneous vegetation during the rainy season (August-November). The rainy season was divided into 6 2-week periods (P1-P6), and environmental conditions (rainy days, rainfall, temperature and humidity) were recorded daily. Five pairs each of tracer kids and tracer lambs raised free of GIN infections were used. Every 15 days, different pairs of kids and lambs were introduced to a 2.2-ha plot and co-grazed with a flock of 30 sheep and 70 goats for a period of 3 weeks. Feeding behaviour of each pair of tracers was measured in weeks 2 and 3. The continuous bite monitoring method was used to estimate total DMI, DMI of plant life forms and DMI from plants of different strata. After each 3-week period, the tracer pair was maintained indoors for 28 days and necropsied on day 29 to recover the worm burden. The feeding behaviour of the tracers was compared between periods (P2-P6) and between kids and lambs. The differences in the worm burdens of the tracers between periods were not associated with total DMI or DMI from plant life forms. Worm burdens were highest during P5 and P6 in tracer kids and lambs (P < 0.05), suggesting a build-up of infective larvae in the tropical deciduous forest (TDF). The lower worm burdens of tracer kids compared with lambs (P < 0.05) seemed to be associated with less low- and more medium-stratum ingestion.
Assuntos
Doenças das Cabras/parasitologia , Infecções por Nematoides/veterinária , Contagem de Ovos de Parasitas/veterinária , Doenças dos Ovinos/parasitologia , Ração Animal , Animais , Fezes , Comportamento Alimentar , Gastroenteropatias/veterinária , Cabras , Enteropatias Parasitárias/parasitologia , Enteropatias Parasitárias/veterinária , Nematoides , Chuva , Estações do Ano , OvinosRESUMO
Rose Bengal@α-cyclodextrin (RB@α-CD) microparticles (µPs) were prepared and the RB inclusion in α-CD was experimentally demonstrated through infrared, UV-VIS absorption spectroscopy and cyclic voltammetry. The RB inclusion in α-CD was theoretically investigated using classical molecular mechanics calculations, the simulation results showing that RB can be included in both the narrow and wide apertures of the α-cyclodextrin ring with configurations exhibiting average binding energies of about 27 kcal mol-1. The prepared RB@α-CD microparticles were characterized through Scanning Electron Microscopy (SEM) and it was demonstrated that they are highly efficient in the photodynamic therapy against a Streptococcus mutans (the main bacteria of cariogenic dental plaque) suspension, as a concentration of RB@α-CD µPs 10 times smaller than the usual concentration of pure RB is still capable to produce significant antibacterial activity.
Assuntos
Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Rosa Bengala/farmacologia , Streptococcus mutans/efeitos dos fármacos , alfa-Ciclodextrinas/química , Biofilmes , Microscopia Eletrônica de Varredura , Tamanho da Partícula , Fármacos Fotossensibilizantes/administração & dosagem , Rosa Bengala/administração & dosagem , Espectrofotometria InfravermelhoRESUMO
Hypovitaminosis D has been identified as a possible new cardiovascular risk factor. However, the results of studies correlating serum vitamin D levels with markers of subclinical atherosclerosis have been conflicting. The aim of this study was to correlate serum levels of 25-hydroxyvitamin D [25(OH)D] with carotid intima-media thickness (C-IMT) and conventional cardiovascular risk factors in Afro-descendants. A cross-sectional analysis was performed on a sample of 382 individuals from a cohort of descendants of African slaves, inhabitants of "Quilombola" communities, with a mean age of 57.79 ±15.3 years, 54.5% of whom were women. Socio-demographic and clinical data were collected and biochemical tests were performed, including serum levels of 25(OH)D by electrochemiluminescence and urinary albumin excretion, evaluated by the albumin/creatinine ratio (ACR) in a spot urine sample. All participants underwent high-resolution ultrasonography for C-IMT measurement. Hypovitaminosis D was defined as serum 25(OH)D levels <30 ng/mL. The mean serum 25(OH)D levels were 50.4±13.5 ng/mL, with a low prevalence of hypovitaminosis D (4.86%). By simple linear correlation, a significant inverse association between 25(OH)D levels and C-IMT (r=-0.174, P=0.001) was observed. However, after multiple linear regression analysis, the significance of the association between serum levels of 25(OH)D and C-IMT measurement was lost (ß=-0.039, P=0.318) and only male gender, age, smoking, systolic blood pressure, glucose and low density lipoprotein (LDL)-cholesterol remained significantly associated with C-IMT. Levels of 25(OH)D were independently and positively associated with HDL-cholesterol and inversely associated with age and ACR. In conclusion, no independent association between 25(OH)D levels and C-IMT was observed in this population. On the other hand, there was an inverse association with albuminuria, a marker of endothelial lesion.
Assuntos
Aterosclerose/sangue , População Negra , Espessura Intima-Media Carotídea , Deficiência de Vitamina D/epidemiologia , Vitamina D/análogos & derivados , Adolescente , Adulto , Idoso , Albuminúria , Aterosclerose/etiologia , Pressão Sanguínea , Brasil/epidemiologia , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Fatores Socioeconômicos , Vitamina D/sangue , Adulto JovemRESUMO
Gastrointestinal nematodes (GINs) cause considerable economic losses in grazing goat herds. At present, GIN control cannot rely on conventional anthelmintic (AH) drugs because parasites have developed resistance against such drugs. Thus, alternative control methods are being sought to reduce the dependence on AH. Many tannin-rich plants exhibit AH activity and may be used as alternatives for GIN control. Mimosa caesalpiniifolia is a tannin-rich shrub consumed by small ruminants in Brazil. This study evaluated the in vivo AH effect of M. caesalpiniifolia leaf powder supplementation on GIN egg fecal excretion and worm burden in goats. Plant leaves were harvested, dried and ground to obtain a powder. Twenty-four castrated male goats, aged six to eight months, with a mean body weight of 15.0⯱â¯2.5â¯kg were used in the experiment. Animals were infected orally with 16,000 larvae comprising 50% Haemonchus spp., 41% Trichostrongylus spp. and 9% Oesophagostomum spp. Once the infection was patent, the goats were distributed into four groups of six animals. The control group received concentrate without condensed tannins (CTs) and did not receive any drench against GINs. The monepantel group received concentrate without CTs and were drenched once with monepantel. The other two groups received the M. caesalpiniifolia leaf powder in two periods of seven consecutive days (days 1-7 and 14-21), with one of the groups also receiving 10â¯g of polyethyleneglycol (PEG)/day. The animals were weighed weekly, and individual fecal eggs counts (FECs) were performed daily. After 28 days, the animals were humanly slaughtered, and the worm burden was estimated. Although live weight gain and FECs did not differ among the groups (Pâ¯>â¯0.05), post-mortem worm counts showed a reduction in Haemonchus contortus adult worm burden (57.7%) in goats of the CT group compared to control goats (Pâ¯<â¯0.05). The addition of PEG did not diminish AH activity in the CTâ¯+â¯PEG group (66.9% reduction compared to the control). No AH effect against other GIN species was found. The result for the addition of PEG suggested that the observed AH activity was associated with plant secondary compounds, as opposed to CTs. As expected, no AH effect against Oesophagostomum columbianum was found for the monepantel group showed. Thus, feeding dry leaves of M. caesalpiniifolia represent a promising alternative for the control of GIN infections in goats.
Assuntos
Anti-Helmínticos/uso terapêutico , Hemoncose/tratamento farmacológico , Hemoncose/veterinária , Haemonchus/efeitos dos fármacos , Mimosa/anatomia & histologia , Folhas de Planta/química , Aminoacetonitrila/administração & dosagem , Aminoacetonitrila/análogos & derivados , Aminoacetonitrila/uso terapêutico , Ração Animal/análise , Animais , Brasil/epidemiologia , Suplementos Nutricionais , Fezes/parasitologia , Trato Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/parasitologia , Doenças das Cabras/tratamento farmacológico , Doenças das Cabras/epidemiologia , Doenças das Cabras/parasitologia , Cabras/parasitologia , Hemoncose/epidemiologia , Hemoncose/parasitologia , Contagem de Ovos de Parasitas , Proantocianidinas/administração & dosagem , Proantocianidinas/química , Proantocianidinas/uso terapêuticoRESUMO
We aimed to evaluate the effect of two vitrification methods on the morphology and functionality of vitrified feline preantral follicles. Feline ovarian tissue was vitrified with EG + trehalose combined or not with dimethyl sulphoxide (DMSO), using two different techniques (open or closed systems). Morphology, developmental capacity and mRNA expression of markers for follicle survival and quality were assessed before and after in vitro culture (IVC). Both vitrification and culture media were serum-free. Vitrification of feline ovarian tissue from five adult domestic cats was performed with EG + trehalose combined or not with DMSO. Two systems were used: the open system solid-surface vitrification (SSV) and the closed system ovarian tissue cryosystem (OTC). Histological analysis of follicle integrity showed that the percentages of normal follicles in previously vitrified ovarian fragments decreased after 7 days of in vitro culture (IVC), independently of the protocol used. Although follicular activation was observed by Ki-67 labelling, this was accompanied by extensive follicular degeneration as detected by a 3-4-fold decrease in follicular density. Remarkable follicle activation was observed in the ovarian tissue vitrified using OTC and subjected to IVC, probably due to a higher rate of degeneration of developing follicles. Even with such follicular loss, the results are promising for the combination of EG + DMSO + trehalose in a serum-free medium when applying the SSV method, with this approach resulting in the highest rates of normal developing follicles (19%) after 7 days IVC, together with granulosa cells proliferating at the same rate observed in fresh tissue.
Assuntos
Gatos , Criopreservação/veterinária , Ovário/fisiologia , Vitrificação , Animais , Apoptose , Crioprotetores/farmacologia , Dimetil Sulfóxido/farmacologia , Ecossistema , Etilenoglicol/farmacologia , FemininoRESUMO
We investigated the association between an aggrecan gene (ACAN) polymorphism and lumbar disc herniation (LDH). This was a case-control study with quinquennial age and gender groups. The study comprised 119 men and women aged between 20 and 60 from Goiânia (Brazil). Of these, 39 were allocated to the case group (Ca) and 80 to the control group (Ct). We gathered sociodemographic and clinical data, and peripheral blood samples. DNA was isolated for genotyping the ACAN variable number tandem repeat (VNTR) via conventional polymerase chain reaction (PCR). Data were statistically analyzed using the chi-square test, multiple comparison analysis, the Student t-test, and odds ratios, with a level of significance set at 5% (P ≤ 0.05). The groups were homogenous in terms of sociodemographic, anthropometric, and life style variables. The allele score for the ACAN VNTR was significantly lower in volunteers with LDH; the A22 allele was significantly more prevalent in this same group; the Ca group presented greater frequency of short alleles A13-A25, whereas the Ct group presented a higher frequency of long alleles. However, this difference was not statistically significant. In both groups, the most common alleles were A28, A27, and A29, and the A26/A26 genotype was significantly more common in the Ca group. The results showed an association between short alleles and LDH among the investigated adults (Ca), corroborating the hypothesis that aggrecan with shorter repeat lengths can lead to a reduction in the physiological proteoglycan function of intervertebral disc hydration and, consequently, increased individual susceptibility to LDH.
Assuntos
Agrecanas/genética , Degeneração do Disco Intervertebral/genética , Deslocamento do Disco Intervertebral/genética , Repetições Minissatélites , Adulto , Povo Asiático/genética , Estudos de Casos e Controles , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
RESUMO A resistência de fungos do gênero Candida aos fármacos químicos tem lançado o desafio de se identificar novas substâncias que possuam atividade antibiótica ou venham a modular o efeito de produtos atualmente usados contra candidíase. O presente estudo avaliou a atividade antifúngica do óleo essencial de Lippia sidoides Cham. e do timol, sobre cepas de Candida. Inicialmente os produtos foram testados frente a 16 cepas fúngicas pela técnica de difusão em meio sólido, o que permitiu selecionar linhagens para continuidade da pesquisa. Com as linhagens de Candida krusei (CK LMBM 01, CK LMBM 02), Candida albicans (CA LM 62) e Candida tropicalis (CT LM 20), procedeu-se, por microdiluição em caldo, a determinação da Concentração Inibitória Mínima (CIM) e em meio sólido, a Concentração Fungicida Mínima (CFM) dos produtos foi identificada. O microcultivo das leveduras em meio empobrecido foi realizado para verificação de alterações morfológicas e, além disso, uma análise da composição química do óleo foi realizada por Cromatografia Gasosa acoplada à espectrometria de massas (CG-EM). Nesta análise, o constituinte majoritário foi o timol (84,95%), seguido de compostos como p-cimeno e Éter metil carvacrol, entre outros. A CIM do óleo essencial de Lippia sidoides Cham. frente às cepas variou entre 64 e 256 μg/mL, enquanto a CIM do timol foi estabelecida entre 32 e 64 μg/mL. A CFM do óleo essencial foi determinada entre 128 e 512 μg/mL e para o timol foram encontrados valores entre 64 e 128 μg/mL. Em relação à análise micromorfológica, verificada nas concentrações de CIM e CIM x 2, o óleo essencial inibiu o dimorfismo das cepas CK 01 e CT 20 na CIM e quando foi ensaiado o timol, este, na CIM, impediu a transição morfológica das cepas CK 01 e CA 62. Uma redução da morfogênese também foi obsevada na cepa CT 20, porém apenas em CIM x 2 e de forma mais discreta. Os resultados enaltecem o potencial antifúngico de L. sidoides e de seu composto majoritário timol tanto no combate à Candida quanto na neutralização de um dos fatores de virulência, a capacidade invasiva por formação de hifas e pseudohifas verificado na condição patogênica da candidíase. Estes dados são promissores e poderão incentivar futuras pesquisas sobre os aspectos fitoquímicos, toxicológicos e farmacológicos tanto do óleo essencial de Lippia sidoides como também de seus componentes químicos.
ABSTRACT The resistance of the Candida against drugs has been a challenge to the discovery of new substances with antimicrobial or modulatory effects that could be used against the cadidiasis. This work evaluated the antifungal activity of the essential oil of Lippia sidoides Cham. and of the Thymol against Candida strains. The products were tested towards 16 strains of Candida using the diffusion method, which allowed to select the strains in order to proceed with the research. The strains of Candida krusei (CK LMBM 01, CK LMBM 02), Candida albicans (CA LM 62) and Candida tropicalis (CT LM 20) were assayed by the microdilution method so that the Minimal Inhibitory Concentration (MIC) and the Minimal Fungicide Concentration (MFC) could be determined. The morphogenesis of the Candida was evaluated using poor environment in order to observe morphological changes. The composition of the essential oil was determined by GC-MS. The main compound observed was the thymol (84.95%). The MIC of the essential oil of L. sidoides and Thymol ranged between 64 to 256 μg/mL, and between 32 to 64 μg/mL respectively. The MFC of the essential oil and the thymol varied between 128 to 512 μg/mL and 64 to 128 μg/mL respectively. The morphogenesis of different Candida strains was inhibited in the MIC and MICx2 to the essential oil and thymol. The results indicated the antifungal potential of the L. sidoides and of the Thymol due to the inhibition of the invasive capacity, one of the most important virulence factors for the candidiasis` development. These results are promising to new researches about the phytochemical, toxicological and pharmacological aspects of the essential oil of L. sidoides and its phytochemical compounds.
Assuntos
Candida/química , Óleos Voláteis/análise , Lippia/classificação , Timol/análise , Virulência , AntifúngicosRESUMO
A leishmaniose e a tripanossomíase americana fazem parte de um grupo de doenças tropicais endêmicas, especialmente entre as populações pobres. São consideradas doenças negligenciadas por não despertarem interesse da indústria farmacêutica. Atualmente, a quimioterapia é o único tratamento específico disponível para estas doenças, onde os medicamentos utilizados são nifurtimox e benzonidazol. Este trabalho teve como objetivo avaliar a atividade antiparasitária e citotóxica do extrato bruto etanólico e frações de folhas de Piper arboreum, frente a linhagens de Leishmania brasiliensis e de Trypanosoma cruzi. Após a preparação do extrato etanólico bruto e as suas respectivas frações, testes in vitro foram realizados para avaliar atividade antiparasitária frente a T. cruzi, utilizado o clone epimastigota CL-B5 e contra L.brasiliensis foram utilizadas formas promastigotas. O ensaio de citotoxicidade foi realizado com linhagens de fibroblastos NCTC929. Os resultados indicaram que as amostras apresentaram toxidade elevada, com exceção da fração de acetato de etila. Já os resultados da atividade antiparasitária indicaram uma maior atividade da fração hexânica observando a morte de 62% e 51% de células epimastigotas e de 100% e 92% de células promastigotas, respectivamente. Através dos resultados conclui-se que a P. arboreum pode ser considerada uma fonte alternativa de produtos naturais com atividade contra T. cruzi e L. brasiliensis...
Leishmaniasis and South American trypanosomiasis (Chagas disease) are part of a group of related human diseases endemic in tropical regions, especially among the poor. They are referred to as neglected diseases, because of a lack of interest on the part of the pharmaceutical industry. Today, chemotherapy is the only specific treatment against these diseases, the most frequently used drugs being nifurtimox and benznidazole. The aim of this study was to assess the antiparasitic and cytotoxic activity of the crude ethanolic leaf extract of Piper arboreum and its fractions against strains of Leishmania brasiliensis and Trypanosoma cruzi. After preparing the crude ethanolic extract and fractions, in vitro tests were performed to assay antiparasitic activity against T. cruzi, using the epimastigote clone CL-B5, while against L. brasiliensis, promastigote forms were used. The cytotoxicity assay was performed with a fibroblast cell line, NCTC 929. The results indicate that the samples possessed high toxicity, except for the ethyl acetate fraction. The results for antiparasitic activity showed highest activity in the hexane fraction, which was lethal to 62% and 51% of the epimastigotes and 100% and 92% of the promastigotes, at 500 and 100µg/mL, respectively. From these results, it is concluded that P. arboreum can be considered an alternative source of natural products with activity against T. cruzi and L.brasiliensis...
Assuntos
Leishmania braziliensis , Piperaceae/toxicidade , Trypanosoma cruziRESUMO
Ovarian fragments were exposed to 0.5 M sucrose and 1 M ethylene glycol (freezing solution; FS) with or without selenium or Trolox. Histological and ultrastructural analyses showed that the percentages of normal follicles in control tissue and in tissue after exposure to FS + 50 µM Trolox were similar. Trolox prevented endoplasmic reticulum (ER)-related vacuolization, which is commonly observed in oocytes and stromal tissue after exposure to FS. From the evaluated stress markers, superoxide dismutase 1 (SOD1) was up-regulated in ovarian tissue exposed to FS + 10 ng/ml selenium. Ovarian fragments were subsequently frozen-thawed in the presence of FS with or without 50 µM Trolox, followed by in vitro culture (IVC). Antioxidant capacity in ovarian fragments decreased after freeze-thawing in Trolox-free FS compared with FS + 50 µM Trolox. Although freezing itself minimized the percentage of viable follicles in each solution, Trolox supplementation resulted in higher rates of viable follicles (67 %), even after IVC (61 %). Furthermore, stress markers SOD1 and ERp29 were up-regulated in ovarian tissue frozen-thawed in Trolox-free medium. Relative mRNA expression of growth factors markers was evaluated after freeze-thawing followed by IVC. BMP4, BMP5, CTGF, GDF9 and KL were down-regulated independently of the presence of Trolox in FS but down-regulation was less pronounced in the presence of Trolox. Thus, medium supplementation with 50 µM Trolox prevents ER stress and, consequently, protects ovarian tissue from ER-derived cytoplasmic vacuolization. ERp29 but not ERp60, appears to be a key marker linking stress caused by freezing-thawing and cell vacuolization.
Assuntos
Cebus/metabolismo , Cromanos/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Congelamento , Ovário/efeitos dos fármacos , Ovário/patologia , Vitamina E/análogos & derivados , Animais , Calreticulina/metabolismo , Crioprotetores/farmacologia , Feminino , Proteínas de Choque Térmico HSP70/metabolismo , Ovário/metabolismo , Ovário/ultraestrutura , Superóxido Dismutase/metabolismoRESUMO
Allergic airway inflammation is attenuated by oral tolerization (oral exposure to allergen, followed by conventional sensitization and challenge with homologous antigen), which decreases airway allergen challenge-induced eosinophilic infiltration of the lungs and bone marrow eosinophilia. We examined its effects on bone marrow eosinophil and neutrophil production. Mice of wild type (BP-2, BALB/c, and C57BL/6) and mutant strains (lacking iNOS or CD95L) were given ovalbumin (OVA) or water (vehicle) orally and subsequently sensitized and challenged with OVA (OVA/OVA/OVA and H2O/OVA/OVA groups, resp.). Anti-OVA IgG and IgE, bone marrow eosinophil and neutrophil numbers, and eosinophil and neutrophil production ex vivo were evaluated. T lymphocytes from OVA/OVA/OVA or control H2O/OVA/OVA donors were transferred into naïve syngeneic recipients, which were subsequently sensitized/challenged with OVA. Alternatively, T lymphocytes were cocultured with bone marrow eosinophil precursors from histocompatible sensitized/challenged mice. OVA/OVA/OVA mice of the BP-2 and BALB/c strains showed, relative to H2O/OVA/OVA controls, significantly decreased bone marrow eosinophil counts and ex vivo eosinopoiesis/neutropoiesis. Full effectiveness in vivo required sequential oral/subcutaneous/intranasal exposures to the same allergen. Transfer of splenic T lymphocytes from OVA/OVA/OVA donors to naive recipients prevented bone marrow eosinophilia and eosinopoiesis in response to recipient sensitization/challenge and supressed eosinopoiesis upon coculture with syngeneic bone marrow precursors from sensitized/challenged donors.
Assuntos
Células da Medula Óssea/imunologia , Hipersensibilidade/imunologia , Imunidade Inata , Pulmão/imunologia , Administração Oral , Alérgenos/administração & dosagem , Alérgenos/imunologia , Animais , Eosinófilos/patologia , Hematopoese/imunologia , Humanos , Hipersensibilidade/patologia , Inflamação/imunologia , Inflamação/patologia , Pulmão/patologia , Camundongos , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Linfócitos T/imunologiaRESUMO
The urinary steroid profile has been used in clinical endocrinology for the early detection of enzyme deficiencies. In the field of doping, its evaluation in urine samples is used to diagnose the abuse of substances prohibited in sport. This profile is influenced by sex, age, exercise, diet, and ethnicity, among others; laboratories own reference ranges might compensate for ethnic differences among population and inter-laboratory biases. This paper shows the reference ranges obtained in the Antidoping Laboratory of Havana for the following steroid profile parameters: ten androgens (testosterone, epitestosterone, androsterone, etiocholanolone, 5α-androstan-3α,17ß-diol, 5ß-androstan-3α,17ß-diol, dehydroepiandrosterone, epiandrosterone, 11ß-hydroxyandrosterone and 11ß-hydroxyetiocholanolone), three estrogens (estradiol, estriol and estrone), two pregnanes (pregnanediol and pregnanetriol) and two corticosteroids (cortisol and tetrahydrocortisol). The urine samples (male: n = 2454 and female: n = 1181) and data obtained are representative of population from Latin-American countries like Cuba, Venezuela, Mexico, Dominican Republic, Guatemala and Chile. Urine samples were prepared by solid-phase extraction followed by enzymatic hydrolysis and liquid-liquid extraction with an organic solvent in basic conditions. Trimethylsilyl derivatives were analyzed by gas chromatography coupled to mass spectrometry. Reference ranges were established for each sex, allowing the determination of abnormal profiles as a first diagnostic tool for the detection of the abuse of androgenic anabolic steroids. The comparison with the Caucasian population confirms that the urinary steroid profile is influenced by ethnicity.
Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Esteroides/urina , Detecção do Abuso de Substâncias/métodos , Corticosteroides/urina , Androgênios/urina , Dopagem Esportivo , Estrogênios/urina , Feminino , Cromatografia Gasosa-Espectrometria de Massas/normas , Hispânico ou Latino , Humanos , América Latina , Masculino , Pregnanodiol/urina , Pregnanotriol/urina , Valores de Referência , Sensibilidade e Especificidade , Detecção do Abuso de Substâncias/normasRESUMO
There is no tradition in studies reporting the effect of exposure to cryoprotectants or simply hypoxia and hypothermia on gene expression in the ovarian tissue and there has been only one study on reference or target genes quantification, and comparisons of normoxic with hypoxic, hypothermic and toxic conditions. Our aim in the present study was to investigate the stability of three reference genes in the ovarian tissue of capuchin monkeys (Cebus apella). To this end, fresh and cryoprotectant-exposed ovarian biopsies were used. Both fresh and exposed ovarian tissues were subjected to total RNA extraction and synthesis of cDNA. cDNA was amplified by real-time polymerase chain reaction (PCR), and GeNorm, BestKeeper and NormFinder software were used to evaluate the stability of glyceraldehyde-2-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyltransferase 1 (HPRT1) and TATA-binding protein (TBP). Results demonstrated that, in the ovarian tissue from capuchin monkeys, HPRT1 and TBP were the most suitable reference genes and thus could be used as parameters to normalize data in future studies. In contrast, GAPDH appeared as the least stable gene among the tested reference genes. In conclusion, HPRT1 and TBP were the most stable reference genes in fresh and cryoprotectant-exposed ovarian tissue from capuchin monkeys.