Anti-snake venom activities of extracts and fractions from callus cultures of Sapindus saponaria

Context: Sapindus saponaria L. (Sapindaceae) bark, root, and fruits are used as sedatives and to treat gastric ulcer and also demonstrate diuretic and expectorant effects. Objective: The anti-snake venom properties of callus of S. saponaria are investigated here for the first time. Materials and methods: In vitro cultivated callus of Sapindus saponaria were lyophilized, and the extracts were prepared with different solvents, before submitting to phytochemical studies and evaluation of the anti-ophidian activity. Crude extracts were fractionated by liquid-liquid partition and the fractions were monitored by thin layer chromatography (TLC). Subsequently, anti-ophidian activities were analyzed toward Bothrops jararacussu Lacerda (Viperidae), B. moojeni Hoge (Viperidae), B. alternates Dumeril (Viperidea) and Crotalus durissus terrificus Lineu (Viperidae) venoms and isolated myotoxins and phospholipase A(2) (PLA(2)). Results: Fractions A1, A2 and the extract in MeOH:H2O (9:1) significantly inhibited the toxic and pharmacological activities induced by snake venoms and toxins, when compared to other extracts and fractions. The lethal, clotting, phospholipase, edema-inducing, hemorrhagic and myotoxic activities were partially inhibited by the different extracts and fractions. TLC profiles of the crude extracts (B and C) and fractions (A1 and A2) showed beta-sitosterol and stigmasterol as their main compounds. Stigmasterol exhibited inhibitory effects on enzymatic and myotoxic activities of PLA(2). Discussion and conclusion: Sapindus saponaria extracts and fractions presented anti-ophidian activity and could be used as an adjuvant to serum therapy or for its supplementation, and in addition, as a rich source of potential inhibitors of enzymes involved in several pathophysiological human and animal diseases.

Skeletal abnormalities in humpback whales Megaptera novaeangliae stranded in the Brazilian breeding ground

Skeletal tissues of 49 humpback whales Megaptera novaeangliae that stranded between 2002 and 2011 along the Abrolhos Bank seashore and its adjacent waters in Brazil were studied. Twelve (24.5%) animals presented pathological changes in one or more bones. Degenerative changes and developmental malformations were most frequent (10.2% each), followed by inflammatory/infectious and traumatic lesions (8.2% each). Infectious diseases led to severe lesions of the caudal vertebrae of 2 whales. In one of these individuals, the lesions involved 6 caudal vertebrae, leading to ankylosis of 3 vertebrae. Degenerative changes were observed in the vertebral columns of 3 animals, involving the joints of 13 ribs of 1 individual, and in the humerus of 1 whale. Traumatic lesions, such as osseous callus in the ribs, were observed in 4 animals. In 1 whale, the rib showed severe osteomyelitis, possibly resulting from the infection of multiple fractures. Developmental abnormalities such as spina bifida on 3 cervical vertebrae of 1 whale, fusion of spinal processes on thoracic vertebrae of 1 individual and fusion of the first 2 ribs unilaterally or bilaterally in 4 animals were found. Chronic infectious conditions found in the axial skeleton may have restrained spinal mobility and had detrimental effects on the general health of the animals, contributing to stranding and death. To our knowledge, this is the first systematic study on skeletal lesions in stranded humpback whales.

Regeneration and characterization of somatic hybrids combining sweet orange and mandarin/mandarin hybrid cultivars for citrus scion improvement

Protoplast fusion between sweet orange and mandarin/mandarin hybrids scion cultivars was performed following the model "diploid embryogenic callus protoplast + diploid mesophyll-derived protoplast". Protoplasts were isolated from embryogenic calli of 'Pera' and 'Westin' sweet orange cultivars (Citrus sinensis) and from young leaves of 'Fremont', Nules', and 'Thomas' mandarins (C. reticulata), and 'Nova' tangelo [C. reticulata x (C. paradisi x C. reticulata)]. The regenerated plants were characterized based on their leaf morphology (thickness), ploidy level, and simple sequence repeat (SSR) molecular markers. Plants were successfully generated only when 'Pera' sweet orange was used as the embryogenic parent. Fifteen plants were regenerated being 7 tetraploid and 8 diploid. Based on SSR molecular markers analyses all 7 tetraploid regenerated plants revealed to be allotetraploids (somatic hybrids), including 2 from the combination of 'Pera' sweet orange + 'Fremont' mandarin, 3 'Pera' sweet orange + 'Nules' mandarin, and 2 'Pera' sweet orange + 'Nova' tangelo, and all the diploid regenerated plants showed the 'Pera' sweet orange marker profile. Somatic hybrids were inoculated with Alternaria alternata and no disease symptoms were detected 96 h post-inoculation. This hybrid material has the potential to be used as a tetraploid parent in interploid crosses for citrus scion breeding.

Somatic embryogenesis from ovaries of sweet orange cv. Tobias

Callogenesis, somatic embryogenesis, and regeneration were obtained from tissues of unfertilized ovaries of sweet orange (Citrus sinensis Osbeck.) cv. Tobias. The influence of two modified basal media, woody plant medium (WPM) and N6 medium, to induce callus formation from pistils was determined. Overall, high frequencies of callogenesis were observed when either medium was used. However, initial culture of explants in WPM medium followed by transfer of callus to N6 medium resulted in higher frequency of callus induction (of 2.30 callus per explant that were larger than 0.5 cm in size), and of subsequent development of embryogenic callus (10%). A total of 125 somatic embryos were obtained. After 6 months of culture, 72% of somatic embryos germinated into plantlets. These plantlets were subsequently micrografted in vitro, and then acclimatized. Ploidy of these plants were determined using flow cytometry and TRAPS molecular markers were used to confirm their maternal origin.

Taxonomic study on the molluscs collected in Marion-Dufresne expedition (MD55) to SE Brazil: Xenophoridae, Cypraeoidea, mitriforms and Terebridae (Caenogastropoda)

The deep-water molluscs collected during the expedition MD55 off SE Brazil have been gradually studied in some previous papers. The present one is focused on samples belonging to caenogastropod taxa Xenophoridae Troschel, 1852, Cypraeoidea Rafinesque, 1815, mitriforms and Terebridae Morch, 1852. Regarding the Xenophoridae, Onustus aquitanus n. sp. is a new species, collected off the littoral of Espirito Santo and Rio de Janeiro, Brazil, 430-637 m depth (continental slope). The main characters of the species include the small size (c. 20 mm), the proportionally wide shell, the white colour, the short peripheral flange, the oblique riblets weakly developed and a brown multispiral protoconch. This appears to be the smallest living species of the family, resembling in this aspect fossil species. In respect to the Cypraeoidea, the following results were obtained: family Cypraeidae Rafinesque, 1815: Erosaria acicularis (Gmelin, 1791) and Luria cinerea (Gmelin, 1791) had the deepest record, respectively 607-620 m and 295-940 m, although the samples were all dead, eroded shells. Family Lamellariidae d'Orbigny, 1841: a total of three lots were collected, provisionally identified as Lamellaria spp. as the samples consist of only vestigial shells; possibly each lot represents a different species. Family Pediculariidae Gray, 1853: a sample of Pedicularia tibia Simone, 2005 was found, expanding the range c. 1000 km southwards, from Ceara to Espirito Santo. Family Ovulidae Fleming, 1822: Pseudosimnia lacrima n. sp., collected off Espirito Santo, 607-620 m depth, is described here and is mainly characterised by its strong biconic outline, small size (c. 7 mm), and a thick peripheral callus. Family Triviidae Troschel, 1863: Cleotrivia antillarum (Schilder, 1922) is recorded for the first time as deep as 620 m, and its distribution expanded from Rio Grande do Norte to Espirito Santo; Dolichupis akangus n. sp. with rounded outline and c. 15 transverse ribs; D. pingius n. sp. with the outer lip expanded posteriorly and c. 10 ribs. In respect to the mitriform neogastropods, the following species are emphasised: family Costellariidae MacDonald, 1860: Vexillum sp., 607-620 m depth; Turricostellaria amphissa n. sp., 295 m depth; T. jukyry n. sp.; T. apyrahi n. sp., both 790-1575 m depth; T. ovir n. sp., 1200 m depth; Nodicostellaria crassa (Simone, 1995), 240-600 m depth, with extension northwards of the range up to Espirito Santo; Austromitra decresca n. sp., 60-105 m depth. Family Mitridae Swainson, 1829: Subcancilla joapyra n. sp., 295 m depth; S. cf. straminea (Adams, 1853), 607-620 m depth. Family Volutomitridae Gray, 1854: Microvoluta corona n. sp., 1500-1575 m depth. Family Mitromorphidae Casey, 1904: Mitromorpha sama n. sp., 607-940 m depth; M. mirim n. sp., 60105 m depth. Regarding the conoidean Terebridae, this paper is a complement of a previous study. It deals with a new species Terebra assu Simone n. sp., from the Abrolhos Bank, 295 m depth, characterised by its narrow outline, yellowish colour, weak sculpture on the last whorls, and a proportionally broad, paucispiral protoconch. A second finding of Terebra alagoensis Lima, Tenorio & Barros, 2007 expands the geographic range from Alagoas to north Espirito Santo. A discussion on the systematics of the "complex Terebra doellojuradoi" in South American coast is also provided, highlighting the improbability of synonymy between T. leptapsis Simone, 1999 and T doellojuradoi Carcelles, 1953. Differences in size, sculpture, spire angulation, aperture, and mainly in protoconch, indicate specific separations. The presently studied terebrids belong to the "complex Terebra doellojuradoi", which encompasses closely related, deep-water, small species, possessing a relatively high degree of endemicity.

A Comparative Analysis between Ultrasonometry and Computer-Aided Tomography to Evaluate Bone Healing

An ultrasonometric and computed-tomographic study of bone healing was undertaken using a model of a transverse mid-shaft osteotomy of sheep tibiae fixed with a semi-flexible external fixator. Fourteen sheep were operated and divided into two groups of seven according to osteotomy type, either regular or by segmental resection. The animals were killed on the 90th postoperative day and the tibiae resected for the in vitro direct contact transverse and axial measurement of ultrasound propagation velocity (UV) followed by quantitative computer-aided tomography (callus density and volume) through the osteotomy site. The intact left tibiae were used for control, being examined in a symmetrical diaphyseal segment. Regular osteotomies healed with a smaller and more mature callus than resection osteotomies. Axial UV was consistently and significantly higher (p?=?0.01) than transverse UV and both transverse and axial UV were significantly higher for the regular than for the segmental resection osteotomy. Transverse UV did not differ significantly between the intact and operated tibiae (p?=?0.20 for regular osteotomy; p?=?0.02 for resection osteotomy), but axial UV was significantly higher for the intact tibiae. Tomographic callus density was significantly higher for the regular than for the resection osteotomy and higher than both for the intact tibiae, presenting a strong positive correlation with UV. Callus volume presented an opposite behavior, with a negative correlation with UV. We conclude that UV is at least as precise as quantitative tomography for providing information about the healing state of both regular and resection osteotomy. (C) 2011 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:10761082, 2012

In vitro organogenesis of Eucalyptus grandis: effects of boron and calcium

The in vitro organogenesis of woody species plays an essential role in the improvement of forest products by providing saplings with high commercial value. Furthermore, mineral nutrition plays an important role in the induction of organogenic responses. The objective of this study was to evaluate the effects of boron and calcium in the organogenesis of nodal segments from seedlings of Eucalyptus grandis growing under in vitro conditions. The concentration of boron and calcium in MS medium was modified to induce organogenic responses in 45-day-old nodal segments used as explants. After 60 days, the fresh weight, dry weight, ratio of fresh and dry weight, relative water content and relative matter content accumulated by the explants were evaluated. The concentrations of boron and calcium in the culture medium influenced the in vitro organogenic control of Eucalyptus grandis. Reduced combinations of boron and calcium induced callus formation and dry matter accumulation in the explants. A boron concentration of 100% (1.10 mg L-1) combined with 100% (119.950 mg L-1) and 200% (239.900 mg L-1) of calcium, and 200% (2.20 mg L-1) of boron combined with 100% (119.950 mg L-1) of calcium allowed the induction of well-developed buds, which can be used for the regeneration of micro-plants.