Phytochemical analysis, toxicity and evaluation of antioxidant and antimicrobial activities of leaves of Dipteryx alata Vogel.

In this study, our objective was to conduct a comprehensive phytochemical analysis, determine toxicity levels, and assess the antioxidant and antimicrobial properties of extracts derived from the leaves of Dipteryx alata Vogel, a native species of the Brazilian cerrado flora. Three distinct extracts were prepared utilizing assisted ultrasound and the Soxhlet apparatus, namely, Ultrasound Crude Extract (UCE), Soxhlet Crude Extract (SCE), and the Soxhlet Ethanol Extract (SEE). The phytochemical analysis revealed the presence of flavonoids, tannins, phytosterols, and saponins in all extracts. Additionally, alkaloids were specifically identified in the SCE and SEE extracts. In the analysis using LC-DAD, the compounds gallic acid, rutin, quercetin, luteolin and kampefrol were determined in higher concentrations in the SCE, followed by the SEE and UCE, respectively. The GC-MS analysis revealed the presence of campesterol, stigmasterol and ß-sitosterol in all extracts, with UCE and SCE showing a higher concentration of ß-sitosterol. SCE showed the highest concentration of all identified compounds. In the analysis of antioxidant activity by DPPH• and ABTS•+, SEE showed greater efficiency (IC50 = 2.98 ± 2.92 and 6.57 ± 0.89 µg/mL, respectively). In the toxicity test with Allium cepa, all extracts stimulated root growth at 50 g/mL; UCE and SEE stimulated root growth at 250 g/mL; and SEE inhibited root growth at 750 g/mL. In the Artemia salina toxicity, all extracts were non-toxic. Antibacterial activity was identified in the microorganisms S. aureus and S. mutans; however, the extracts did not show antifungal action against the strain of C. albicans. The extracts of D. alata have therapeutic potential for applicability in dentistry.

Parkia from Cerrado: phytochemical bioprospection, toxicity and in vitro bioactivities of bark and flower extracts.

Parkia platycephala is the only species of the genus Parkia that is endemic to the brazilian Cerrado and the tree symbol of the state of Tocantins, but there are still few studies regarding its bioprospecting. In this study, we aimed to investigate the phytochemical composition, toxicity and bioactivities of the bark and flower of Parkia platycephala. Hot sequential extractions (Soxhlet) were performed using methanol and hydroethanolic solution (70%), after degreasing the sample (hexane). The presence of flavonoids, tannins, steroids and alkaloids was detected in the preliminary screening. Trilinolein, (Z)-9-octadecenamide, 3-O-methyl-d-glucose were detected by Gas Chromatography coupled to Mass Spectrometry (GC-MS). In the Liquid Chromatography with Diode Array Detector (LC-PDA) analysis, it was detected exclusively ferulic acid (bark) and ellagic acid (flower). The ethanolic extract of the bark (IC50=10.69 ± 0.35 µgmL-1) has an antioxidant potential (DPPH• radical) higher than that of the rutin standard (IC50=15.85 ± 0.08 µgmL-1). All extracts showed excellent anticholinesterase potential (Ellman), with emphasis on the ethanol extract of the flower (IC50 =5.34 ± 0.12 µgmL-1). Regarding toxicity (Artemia salina), the methanolic extract of the bark and the ethanolic extract of the flower presented high and moderate levels, respectively. Such results limit the concentrations of biological activities in this study, however, the antioxidant and anticholinesterase indices fall short of toxicity. The results demonstrated promising antioxidant and anticholinesterase activities of both the bark and the flower of Parkia platycephala.

The monodominance of Acuri in Pantanal formations: allelochemical effects of its leaves and the presence of Acurizais.

The presence of monodominant vegetative formations almost exclusively composed of Acuri palm trees (Attalea phalerata) stands out in some regions of the Pantanal Sul-Mato-Grossense. These formations are generally associated with anthropic, edaphic and/or hydrological factors. However, little is known about the effect of allelopathy on the formation and maintenance of these areas. Herein, we investigated the chemical composition of A. phalerata aqueous leaf extract and the potential allelopathic effects on germination and growth of target L. sativa, L. esculentum and S. obtusifolia species. Thus, extracts at concentrations of 0, 2.5, 5, 10, 15, and 20% were used for germination and growth bioassays with a completely randomised design in a germination chamber and greenhouse. The results showed that the A. phalerata extracts negatively affected the germination speed index and mean germination time of the target species and positively affected seedling length under controlled conditions and were also stimulated in the greenhouse. Thus, the formation of Acurizals can be related to the presence of secondary metabolites in the leaves, in addition to other environmental factors.

The monodominance of Acuri in Pantanal formations: allelochemical effects of its leaves and the presence of Acurizais / A monodominância do Acuri em formações do Pantanal: efeitos aleloquímicos de suas folhas e a presença de acurizais

The presence of monodominant vegetative formations almost exclusively composed of Acuri palm trees (Attalea phalerata) stands out in some regions of the Pantanal Sul-Mato-Grossense. These formations are generally associated with anthropic, edaphic and/or hydrological factors. However, little is known about the effect of allelopathy on the formation and maintenance of these areas. Herein, we investigated the chemical composition of A. phalerata aqueous leaf extract and the potential allelopathic effects on germination and growth of target L. sativa, L. esculentum and S. obtusifolia species. Thus, extracts at concentrations of 0, 2.5, 5, 10, 15, and 20% were used for germination and growth bioassays with a completely randomised design in a germination chamber and greenhouse. The results showed that the A. phalerata extracts negatively affected the germination speed index and mean germination time of the target species and positively affected seedling length under controlled conditions and were also stimulated in the greenhouse. Thus, the formation of Acurizals can be related to the presence of secondary metabolites in the leaves, in addition to other environmental factors.

No Pantanal Sul-Mato-Grossense se destaca, em determinadas regiões, a presença de formações vegetacionais monodominantes, compostas quase que exclusivamente por uma espécie de palmeira, o Acuri (Attalea phalerata). Normalmente estas formações estão associadas a fatores antrópicos, edáficos e/ou hidrológicos. Porém pouco se sabe sobre o efeito da alelopatia na formação e manutenção destas áreas. Desta maneira, objetivou-se estudar a composição química dos extratos aquosos das folhas de A. phalerata e seus possíveis efeitos alelopáticos na germinação e crescimento das espécies-alvo, alface, tomate e fedegoso. Para os bioensaios de germinação e crescimento, foram utilizados extratos nas concentrações de 0, 2,5, 5, 10, 15 e 20%, com delineamento inteiramente casualizado, em câmara de germinação e casa de vegetação. Os resultados demostraram que em condições controladas (câmara de germinação), os extratos de A. phalerata afetaram negativamente o índice de velocidade de germinação e o tempo médio de geminação das espécies-alvo e positivamente, o comprimento das plântulas, que também foram estimuladas em casa de vegetação, indicando a presença de fitormônios. Desta maneira, a formação de Acurizais pode estar relacionada à presença de metabolitos secundários nas folhas, além de outros fatores ambientais.

A communicating branch between the dorsal and superficial ramus of the ulnar nerve (Kaplan's anastomosis): clinical and surgery discussion / Ramo comunicante entre los ramos dorsal y superficial del nervio ulnar (anastomosis de Kaplan): discusión clínica y quirúrgica

The Kaplan´s anastomosis represents the communication between the dorsal and superficial branch of the ulnar nerve distal to ulnar canal. In the present study, a case about this nerve communication, found in a male adult cadaver, is reported. Information about brachial plexus distal anastomosis is fundamental for interpretation of clinical and electrophysiological findings, in order to establish the precise diagnosis of neurological lesions at this level. Its relations with flexor carpi ulnaris muscle's tendon and with pisiform bone exposes it to iatrogenic lesions during surgery.

La anastomosis de Kaplan representa la comunicación entre la rama dorsal y superficial del nervio ulnar distal al canal ulnar. En el presente estudio se informa de un caso de esta comunicación nerviosa, encontrado en un cadáver adulto de sexo masculino. Información sobre el plexo braquial y sus anastomosis distales son fundamentales para la interpretación de los hallazgos clínicos y electrofisiológicos, a fin de establecer el diagnóstico preciso de las lesiones neurológicas a este nivel. Sus relaciones con el tendón del músculo flexor ulnar del carpo, como con el hueso pisiforme, pueden facilitar la ocurrencia de lesiones iatrogénicas durante la cirugía.

Output relations of humeral circumflex arteries and its variations / Relaciones de las arterias circunflejas humerales y sus variaciones

A large range of variability marks the branching pattern of the axillary artery. The knowledge of the anatomical variations and this pattern is essential to diagnostic and therapeutic approaches, including surgery, of the axillary region. The aim of this study was to observe the different possible origins of circumflex humeral arteries and to measure the length and diameter of each vessel. In our study, 24 armpits from adult cadavers (fixed in tamponed formalin 10%) were dissected. The data were analyzed with a digital caliper and the results expressed as Mean ± SD. In majority of specimens, posterior circumflex humeral artery (PCHA) arose from subscapular artery (SSA) (54.16%) and had an average diameter of 3.92±0.41 mm. The anterior circumflex humeral artery was a branch from axillary artery (AA) in the majority of the specimens (62.5%) with an average diameter of 1.83±0.68 mm. Circumflex humeral arteries can arise from SSA, deep brachial artery and AA. The result of this study is an interesting data for origin, length and diameter of these vessels, contributing to the knowledge of these variations occurrence.

Una amplia gama de variabilidad marca el patrón de ramificación de la arteria axilar. El conocimiento de las variaciones anatómicas de este patrón es esencial para enfoques de diagnóstico y terapéuticos, incluyendo la cirugía de la región axilar. El objetivo fue observar los diferentes orígenes posibles de arterias circunflejas humerales y medir la longitud y el diámetro de cada vaso. En el estudio se disecaron 24 axilas de cadáveres adultos (fijados en formalina tamponada al 10%). Los datos se analizaron con un calibrador digital y los resultados se expresaron como Media ± DS. En la mayoría de los especímenes, la arteria circunfleja humeral posterior surgió de la arteria subescapular (ASE) (54,16%) con un diámetro medio de 0,41±3,92 mm. En la mayoría de los especímenes (62,5%), la arteria circunfleja humeral anterior era una rama de la arteria axilar (AA) con un diámetro medio de 0,68±1,83 mm. Las arterias circunflejas humerales pueden surgir de la ASE, de la arteria braquial profunda y AA. El resultado de este estudio es un dato interesante para el origen, la longitud y el diámetro de los vasos, lo que contribuye al conocimiento de la ocurrencia de estas variaciones.

Hypotensive and vasorelaxant effects induced by the ethanolic extract of the Mimosa caesalpiniifolia Benth. (Mimosaceae) inflorescences in normotensive rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Caatinga is highly influenced by its seasonality. This species is endemic in the northeastern region, which is rich in plants with pharmacological potential. Many of these plants are used by the population and some of them have confirmed pharmacological properties. Mimosa caesalpiniifolia Benth. (Mimosaceae) is a native plant from northeastern Brazil׳s caatinga, popularly known as sabiá and cascudo. The tea from the inflorescence of this species is used by the population of the semi-arid for the treatment of hypertension, and the utilization of the plant bark for the staunching of bleedings and wound washing in order to prevent inflammation; also, the ingestion of the bark infusion is used in the treatment of bronchitis. However, its pharmacological effects and mechanisms of action have not yet been studied. The aim of the present study was to determine the effect of the ethanolic extract of M. caesalpiniifolia on the cardiovascular system in rats. MATERIAL AND METHODS: In a study for the assessment of the hypotensive effect of the extract, the polyethylene catheters were inserted in the aorta artery and inferior vena cava for the measurement of the arterial pressure and heart rate. When intragastric administration was performed, only one catheter was implanted in the abdominal aorta. In studies for the vasorelaxant activity, mesenteric arterial rings (1-2mm) were used: they were kept in Tyrode׳s solution (95% O2 and 5% CO2) and submitted to tension of 0.75 g/f for 1h. The results were expressed as mean ± S.E.M., significant to the values of p<0.05. RESULTS: The administration of the doses through venous pathway (6.25; 12.5 and 25mg/kg, i.v.) promoted hypotension followed by bradycardia in the higher doses. The pre-treatment with atropine (2mg/kg, i.v.) interrupted both the hypotension and the bradycardia; with hexamethonium, hypotension was reverted and bradycardia was attenuated. While the administration of tea/flowers (25mg/kg i.v.) also promoted a following section of hypotension, a slight increase in heart rate was observed. When administered orally, MC-EtOH/flowers (100mg/kg, v.o.) promoted a decrease in the arterial pressure from 90 min on, without a significant alteration in the heart rate in relation to the control. In the in vitro study, a pharmacological trial was performed with the extracts obtained from parts of the species M. caesalpiifolia (leaves, bark, fruit and inflorescences). Among all extracts tested, the ethanolic extract from the inflorescences (MC-EtOH/flowers) presented higher vasorelaxant potency in relation to the other parts of the plant. Henceforth, MC-EtOH/flowers was used in the sequence. In mesenteric preparations pre-contracted with phenylephrine (10(-5)M), the MC-EtOH/flowers (0.1-750 µg/ml) promoted vasorelaxant effect regardless of the vascular endothelium. MC-EtOH/flowers inhibited the contractions induced by the cumulative addition of phenylephrine (10(-9)-10(-5)mol/l) or CaCl2 (10(-6)-3 × 10(-2)M), in a concentration-dependent way. In contractions induced by S(-)Bay K 8644, a Cav-L activator, the MC-EtOH/flowers promoted concentration-dependent relaxation, corroborating previous results. CONCLUSION: The tea of flowers of M. caesalpiniifolia promotes hypotension and tachycardia, whereas ethanolic extract (MC-EtOH) promotes hypotension and bradycardia involving the participation of the muscarinic and ganglionic pathways, as well as vasorelaxant action involving the Ca(2+) influx inhibition blockade.

Novel rat model of nonunion fracture with vascular deficit.

Nonunion fractures occur frequently in humans, with profound implications (medical and non-medical). Although there are numerous animal models to study pathogenesis and treatment of nonunion fractures, there is apparently the lack of a definitive model for atrophic nonunion fracture. Therefore, the objective was to develop a low-cost rat model of nonunion fracture with a vascular deficit that enabled standardized quantitative analysis of bone growth and regeneration. The model was developed with two surgeries, performed apart. The first involved osteotomy of the femur diaphysis, removal of periosteum and endosteum, isolation of the fracture site using a latex artefact (Penrose drain tube), and reduction of the fracture using an intramedullary pin, whereas the second surgery was to remove the latex artefact. Based on radiographic imaging, micro-CT and histological analyses done 125 days after the fracture was induced, there was clear evidence of atrophic nonunion fracture, without pin migration or specimen loss. Perceived advantages of this model included low cost, ease of reproducibility, lack of specimen loss, and, finally, the potential to assess bone growth and regeneration under poor vascular conditions.

Efeito do extrato aquoso e etanólico do angico preto sobre larvas de Rhipicephalus (Boophilus) microplus / The effect of aqueous and ethanolic extracts from the angico Preto about larvae of Rhipicephalus (Boophilus) microplus

Rhipicephalus (Boophilus) microplus representa um grande problema na bovinocultura e o uso de acaricidas é a medida de controle profilático e terapêutico mais comum contra esses ectoparasitos. Os principais problemas relacionados com essa prática dizem respeito ao desenvolvimento de linhagens resistentes de carrapatos. Assim, objetivou-se determinar o efeito de extratos da casca de Anadenanthera macrocarpa sobre as larvas de R. (B.) microplus, obtidas de um pool de ovos, acondicionadas em tubo de polietileno. De acordo com os resultados, o extrato aquoso na concentração de 8,26mg.mL-1 causou 85% de mortalidade nas primeiras 12 horas. Quanto ao extrato etanólico, observou-se maior mortalidade nas concentrações 12,5; 6,25 e 1,56mg.mL-1, em torno de 84%, percentuais semelhantes ao amitraz. Os controles negativos não apresentaram mortalidade durante o experimento. Assim, tanto o extrato aquoso como o extrato etanólico apresentaram efeito larvicida, embora o extrato etanólico tenha sido mais eficiente para a espécie, podendo ser uma alternativa no controle desse ectoparasito.

Rhipicephalus (Boophilus) microplus represents a major problem in cattle breeding and the use of acaricides is the most common prophylactic and therapeutic control measure against these ectoparasites. The main problems with this practice relate to the development of resistant strains of ticks. Thus, the objective was to determine the effect of bark extracts from Anadenanthera macrocarpa on the larvae of R.(B.) microplus, obtained from a pool of eggs packed in a polyethylene tube. According to the results, the aqueous extract at a concentration of 8.26mg.mL-1caused 85% mortality in the first 12 hours. As for the ethanolic extract, higher mortality of about 84%, a percentage similar to amitraz, was observed at concentrations of 12.5mg.mL-1, 6.25mg.mL-1 and 1.56mg.mL-1. The negative controls showed no mortality during the experiment. Thus, both the aqueous and ethanolic extracts showed larvicidal activity, although the ethanolic extract has been more efficient and could be an alternative to control this ectoparasite.

Efeito do extrato aquoso e etanólico do angico preto sobre larvas de Rhipicephalus (Boophilus) microplus / The effect of aqueous and ethanolic extracts from the angico Preto about larvae of Rhipicephalus (Boophilus) microplus

Rhipicephalus (Boophilus) microplus representa um grande problema na bovinocultura e o uso de acaricidas é a medida de controle profilático e terapêutico mais comum contra esses ectoparasitos. Os principais problemas relacionados com essa prática dizem respeito ao desenvolvimento de linhagens resistentes de carrapatos. Assim, objetivou-se determinar o efeito de extratos da casca de Anadenanthera macrocarpa sobre as larvas de R. (B.) microplus, obtidas de um pool de ovos, acondicionadas em tubo de polietileno. De acordo com os resultados, o extrato aquoso na concentração de 8,26mg.mL-1 causou 85% de mortalidade nas primeiras 12 horas. Quanto ao extrato etanólico, observou-se maior mortalidade nas concentrações 12,5; 6,25 e 1,56mg.mL-1, em torno de 84%, percentuais semelhantes ao amitraz. Os controles negativos não apresentaram mortalidade durante o experimento. Assim, tanto o extrato aquoso como o extrato etanólico apresentaram efeito larvicida, embora o extrato etanólico tenha sido mais eficiente para a espécie, podendo ser uma alternativa no controle desse ectoparasito.(AU)

Rhipicephalus (Boophilus) microplus represents a major problem in cattle breeding and the use of acaricides is the most common prophylactic and therapeutic control measure against these ectoparasites. The main problems with this practice relate to the development of resistant strains of ticks. Thus, the objective was to determine the effect of bark extracts from Anadenanthera macrocarpa on the larvae of R.(B.) microplus, obtained from a pool of eggs packed in a polyethylene tube. According to the results, the aqueous extract at a concentration of 8.26mg.mL-1caused 85% mortality in the first 12 hours. As for the ethanolic extract, higher mortality of about 84%, a percentage similar to amitraz, was observed at concentrations of 12.5mg.mL-1, 6.25mg.mL-1 and 1.56mg.mL-1. The negative controls showed no mortality during the experiment. Thus, both the aqueous and ethanolic extracts showed larvicidal activity, although the ethanolic extract has been more efficient and could be an alternative to control this ectoparasite.(AU)

LC-MS/MS quantitation of plasma progesterone in cattle.

Quantitation of progesterone (P(4)) in biological fluids is often performed by radioimmunoassay (RIA), whereas liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) has been used much less often. Due to its autoconfirmatory nature, LC-MS/MS greatly minimizes false positives and interference. Herein we report and compare with RIA an optimized LC-MS/MS method for rapid, efficient, and cost-effective quantitation of P(4) in plasma of cattle with no sample derivatization. The quantitation of plasma P(4) released from three nonbiodegradable, commercial, intravaginal P(4)-releasing devices (IPRD) over 192 h in six ovariectomized cows was compared in a pairwise study as a test case. Both techniques showed similar P(4) kinetics (P > 0.05) whereas results of P(4) quantitation by RIA were consistently higher compared with LC-MS/MS (P < 0.05) due to interference and matrix effects. The LC-MS/MS method was validated according to the recommended analytical standards and displayed P(4) limits of detection (LOD) and quantitation (LOQ) of 0.08 and a 0.25 ng/mL, respectively. The high selective LC-MS/MS method proposed herein for P(4) quantitation eliminates the risks associated with radioactive handling; it also requires no sample derivatization, which is a common requirement for LC-MS/MS quantitation of steroid hormones. Its application to multisteroid assays is also viable, and it is envisaged that it may provide a gold standard technique for hormone quantitation in animal reproductive science studies.

Specific immune responses but not basal functions of B and T cells are impaired in aged mice.

Senescence is characterized by several alterations in the immune system. Such modifications can be found in lymphoid organs as well as in the cellular components of the immune system. Several reports have suggested that immune dysfunction can affect both T and B cells, but T cells have been shown to be more susceptible to the effects of aging. B cell function may also be altered with reduction in germinal center formation, antibody response, and affinity maturation of antibodies. Herein we showed that although antigen-specific antibody response to a soluble antigen declines in 18-month old mice, total levels of serum antibodies as well as frequencies of spleen and bone marrow antibody-producing cells are increased in aged mice. In addition, proliferative response of non-stimulated spleen T cells from aged mice were augmented and insensitive to increasing doses of concanavalin A stimulation as compared to young mice that showed a typical dose-dependent response to mitogen stimulation in vitro. These data suggest that the higher activation mode of B and T cells in senescent mice is a result of an increased frequency of cells committed to previous antigenic experiences and with poor ability to respond to novel antigenic challenges.

Efeito anti-helmíntico dos extratos aquosos e etanólicos da Annona squamosa L. (fruta-do-conde) sobre o nematóide Ascaridia galli / Anthelmintic effect of aqueous and ethanolic extracts from Annona squamosa L. (sweetsop) on the nematode Ascaridia galli

As plantas são fontes importantes de produtos naturais biologicamente ativos. Dentre as plantas usadas na medicina popular a Anonna squamosa conhecida como fruta-do-conde é citada como tendo várias ações medicinais, dentre elas a atividade inseticida e anti-helmíntica. Dentro desta perspectiva, objetivou-se determinar a atividade anti-helmíntica dos extratos aquosos (EA) e etanólicos (EE) das folhas da fruta-do-conde sobre o nematóide de aves Ascaridia galli, in vitro e in vivo. No primeiro, os nematóides foram colocados em placa de Petri contendo diferentes concentrações dos extratos e no segundo foram utilizadas seis galinhas poedeiras por grupo, as quais foram administrados10 mL Kg-1 dos extratos. No teste in vitro o EA da A. squamosa nas concentrações 2,4 e 9,6 mg mL-1 foi capaz de matar 63,33 por cento e 53,33 por cento dos nematóides, respectivamente. O EE não produziu efeito significativo. No teste in vivo, o percentual de eliminação do EA foi de 39 por cento e do EE de 20 por cento. Estes dados sugerem que neste caso a substância responsável pela mortalidade dos parasitos esteja em maior concentração na fração aquosa. Desta maneira, acredita-se que o EA de A. squamosa apresenta uma atividade anti-helmíntica potencial sobre o A. galli.

Plants are important sources of biologically active natural products. Among the plants used in popular medicine, Annona squamosa, known as sweetsop, is reported to have several medicinal actions such as insecticidal and anthelmintic activity. Therefore, this study aimed to assess the anthelmintic activity of aqueous (AE) and ethanolic (EE) extracts from sweetsop leaves on the chicken roundworm Ascaridia galli, both in vitro and in vivo. In the former, nematodes were placed on a Petri plate containing different concentrations of the extracts; in the in vivo test, six egg-laying chickens per group received 10 mL Kg-1 of the extracts. In vitro results indicated that A. squamosa AE at the concentrations 2.4 and 9.6 mg mL-1 could kill 63.33 percent and 53.33 percent nematodes, respectively. However, EE did not have any significant effect. According to the in vivo test, the elimination percentage for AE was 39 percent and for EE, 20 percent. These data suggest that the substance responsible for parasite mortality was present at a higher concentration in the aqueous fraction. Thus, A. squamosa AE is believed to have a potential anthelmintic activity on A. galli.