Physico-chemical analysis of Albian (Lower Cretaceous) amber from San Just (Spain) : implications for palaeoenvironmental and palaeoecological studies

Amber from a Lower Cretaceous outcrop at San Just, located in the Eastern Iberian Peninsula (Escucha Formation, Maestrat Basin), was investigated to evaluate its physico-chemical properties. Thermogravimetric (TG) and Differential Thermogravimetric (DTG) analyses, infra-red spectroscopy, elemental and C-isotope analyses were performed. Physico-chemical differences between the internal light nuclei and the peripheral darker portions of San Just amber can be attributed to processes of diagenetic alteration that preferentially took place in the external amber border colonized by microorganisms (fungi or bacteria) when the resin was still liquid or slightly polymerized. δ13Camber values of different pieces of the same sample, from the nucleus to the external part, are remarkably homogeneous, as are δ13Camber values of the darker peripheral portions and lighter inner parts of the same samples. Hence, neither invasive microorganisms, nor diagenetic alteration changed the bulk isotopic composition of the amber. δ13C values of different amber samples range from -21.1‰ to -24‰, as expected for C3 plant-derived material. C-isotope analysis, coupled to palaeobotanical, TG and DTG data and infra-red spectra, suggests that San Just amber was exuded by only one conifer species, belonging to either the Cheirolepidiaceae or Aracauriaceae, coniferous families probably living under stable palaeoenvironmental and palaeoecological conditions

An Efficient Vector System to Modify Cells Genetically

The transfer of foreign genes into mammalian cells has been essential for understanding the functions of genes and mechanisms of genetic diseases, for the production of coding proteins and for gene therapy applications. Currently, the identification and selection of cells that have received transferred genetic material can be accomplished by methods, including drug selection, reporter enzyme detection and GFP imaging. These methods may confer antibiotic resistance, or be disruptive, or require special equipment. In this study, we labeled genetically modified cells with a cell surface biotinylation tag by co-transfecting cells with BirA, a biotin ligase. The modified cells can be quickly isolated for downstream applications using a simple streptavidin bead method. This system can also be used to screen cells expressing two sets of genes from separate vectors

Riqueza, abundância relativa e hábitat de reprodução de espécies de anuros terrestres na região do Triângulo Mineiro, bioma Cerrado, sudeste do Brasil

Based on field observations and pitfall sampling, we determined the species richness, relative abundance, and reproductive habitat of terrestrial frogs in three municipalities in the Triângulo Mineiro region, south Cerrado biome, in southeastern Brazil. We found thirty-two species of terrestrial frogs, belonging to the families Brachycephalidae, Bufonidae, Cycloramphidae, Dendrobatidae, Leiuperidae, Leptodactylidae and Microhylidae. Most of the species were found in open areas and reproduced in human-generated environments, such as artificial lakes (10 species) and ponds (14 species). Dominance was high, with Physalaemus cuvieri Fitzinger, 1826 (Leiuperidae) representing 48% of sampled frogs. A larger number of individuals was captured in the wet season, when most of the species were reproducing. Compared to other areas of Cerrado biome, the Triângulo Mineiro sites presented a larger number of species, which may be attributed to the larger sampled area and greater sampling effort, lower altitude and presence of human generated habitats. The richness of terrestrial frogs was also larger than that in some forested localities in southeastern Brazil, indicating that the number of species cannot be explained only by precipitation and type of vegetation cover. The greater abundance of individuals during the wet season may be related to a greater movement of adults to breeding sites and to juvenile recruitment/dispersion. The heterogeneity of environments in the Cerrado biome, including its several isolated highlands, contributes to its high (local and regional) diversity of frogs.Neste estudo foram determinados a riqueza, abundância relativa e hábitat de reprodução de anuros terrestres em três municípios do Triângulo Mineiro, sul do Bioma Cerrado, sudeste do Brasil, baseados em observações de campo e armadilhas de interceptação e queda. Foram encontradas 32 espécies pertencentes às famílias Brachycephalidae, Bufonidae, Cycloramphidae, Dendrobatidae, Leiuperidae, Leptodactylidae e Microhylidae. Muitas dessas espécies foram encontradas em áreas abertas e se reproduziram em ambientes artificiais gerados por ação antrópica, tais como lagos (10 espécies) e poças (14 espécies). Physalaemus cuvieri Fitzinger, 1826 (Leiuperidae) foi a espécie dominante, representando 48% do total amostrado. Um grande número de indivíduos de diferentes espécies foi capturado na estação chuvosa, quando muitas das espécies estavam reproduzindo. As áreas amostradas na região do Triângulo Mineiro apresentaram um maior número de espécies quando comparadas com outras áreas do bioma Cerrado, o que pode ser atribuído à maior área amostrada, ao maior esforço de coleta, baixa altitude e presença de hábitats gerados por ação antrópica. A riqueza de anuros terrestres também foi maior do que àquela encontrada em localidades florestadas no sudeste do Brasil, indicando que o número de espécies não pode ser explicado somente pela precipitação e tipo de cobertura vegetal da área. A grande abundância de indivíduos durante a estação chuvosa pode estar relacionada ao maior movimento de adultos para hábitats de reprodução e ao recrutamento e dispersão de juvenis. A heterogeneidade dos ambientes no bioma Cerrado, incluindo algumas de suas áreas com maiores altitudes, contribui para a alta diversidade (local e regional) de espécies de anuros

Produção de clamidósporos de Pochonia chlamydosporia em diferentes substratos

Clamidósporos são estruturas de sobrevivência do fungo nematófago Pochonia chlamydosporia. Objetivou-se, com este trabalho, avaliar diferentes substratos, teores de água e formas de inóculo para a produção in vitro de clamidósporos de P. chlamydosporia. Inicialmente, testaram-se os substratos grãos de milho triturado, grãos de arroz e casca de café e os tipos de inóculo meio líquido concentrado ou diluído (1:40) e discos de cultura, colonizados por P. chlamydosporia. Posteriormente, testou-se o substrato grãos de milho triturado suplementado ou não com caldo de batata com os inóculos do fungo nas formas de discos de micélio ou suspensão aquosa. As maiores produções de clamidósporos g-1 de substrato foram obtidas nos substratos grãos de milho triturado e casca de café, e as melhores formas de inóculo foram meio líquido diluído (1:40) e discos de micélio. A suplementação do substrato grãos de milho triturado com caldo de batata não aumentou a produção de clamidósporos do fungo e a melhor forma de inóculo do fungo foi a de discos de micélio. Além disso, testaram-se os substratos canjica, grãos de arroz, casca de café e fibra de coco, umedecidos com diferentes quantidades de água; o substrato casca de café umedecido por diferentes períodos; a casca de café enriquecida ou não com sacarose e farinha de arroz e o substrato grãos de arroz esterilizado no forno micro-ondas com diferentes quantidades de água. Apenas o substrato grãos de arroz, em todas as quantidades de água testadas, apresentou maior produção de clamidósporos. Não houve diferença na produção de clamidósporos no substrato casca de café umedecido por diferentes períodos e apenas quando a casca de café foi enriquecida com farinha de arroz, apresentou maior média de clamidósporos g-1 de substrato. Todos os tratamentos apresentaram grande produção de clamidósporos g-1 quando o substrato utilizado foi o de grãos de arroz tratados no forno de micro-ondas. O melhor meio de cultivo de P. chlamydosporia para a produção de clamidósporos foi o substrato contendo grãos de arroz.Chlamydospores are survival structures of the nematophagous fungus Pochonia chlamydosporia. The objective of this study was to evaluate different substrates, different contents of water and kinds of inoculum for the production of Pochonia chlamydosporia chlamydospores. The substrates evaluated were: milled maize, rice grains and coffee husk; all of them were inoculated with disks of culture, concentrated liquid media or diluted liquid media (1:40) colonized by P. chlamydosporia. Besides, the substrates milled maize, supplemented or not with potato broth, and inoculated with fungus disks or aqueous suspension were evaluated. Milled maize and coffee husk were the best substrates for chlamydospores production. The best inoculum kinds were disks of culture and diluted liquid media (1:40) colonized by P. chlamydosporia. The supplementation of milled maize with potato broth did not improve the production of chlamydospores, and the best inoculum form was disks of fungus. Moreover, it was also studied the substrates rice grains, coffee husk and coconut fibers with different rates of water; the substrate coffee husk moistened by different periods; the supplementation of coffee husk with rice flour or dextrose; and the substrate grains of rice sterilized in microwave oven with different rates of water. The substrate grains of rice, in all of the amounts of water tested showed increase in production of chlamydospores. There was no difference in the chlamydospores production of the coffee husk substrate, when moistened by different periods of time and only when it was supplemented with rice flour it showed higher mean of clamydospores per gram of substrate. All treatments evaluated showed higher production of chlamydospores in the substrate grains of rice treated in the microwave oven. The best substrate for the production of chlamydospores of P. chlamydosporia was the rice grains

From sleep spindles of natural sleep to spike and wave discharges of typical absence seizures: is the hypothesis still valid?

The temporal coincidence of sleep spindles and spike-and-wave discharges (SWDs) in patients with idiopathic generalized epilepsies, together with the transformation of spindles into SWDs following intramuscular injection of the weak GABAA receptor (GABAAR) antagonist, penicillin, in an experimental model, brought about the view that SWDs may represent ‘perverted’ sleep spindles. Over the last 20 years, this hypothesis has received considerable support, in particular by in vitro studies of thalamic oscillations following pharmacological/genetic manipulations of GABAARs. However, from a critical appraisal of the evidence in absence epilepsy patients and well-established models of absence epilepsy it emerges that SWDs can occur as frequently during wakefulness as during sleep, with their preferential occurrence in either one of these behavioural states often being patient dependent. Moreover, whereas the EEG expression of both SWDs and sleep spindles requires the integrity of the entire cortico-thalamo-cortical network, SWDs initiates in cortex while sleep spindles in thalamus. Furthermore, the hypothesis of a reduction in GABAAR function across the entire cortico-thalamo-cortical network as the basis for the transformation of sleep spindles into SWDs is no longer tenable. In fact, while a decreased GABAAR function may be present in some cortical layers and in the reticular thalamic nucleus, both phasic and tonic GABAAR inhibitions of thalamo-cortical neurons are either unchanged or increased in this epileptic phenotype. In summary, these differences between SWDs and sleep spindles question the view that the EEG hallmark of absence seizures results from a transformation of this EEG oscillation of natural sleep

Free amino acid profile of Bubalus bubalis L. meat from the Campania region

ABSTRACT In this study, we determined the amount of carnosine and anserine in water buffalo meat without hanging treatment and the free amino acid profile by using amino acid analyser with post-column ninhydrin derivatization procedure. The main free amino acids present in samples were glutamic acid (>60 mg/100 g), followed by alanine, glycine, and arginine. Other protein amino acids were detected in minor amounts (less than 2 mg/100 g). Among the non-protein amine-containing compounds, taurine and urea were the most abundant. The analysis showed that 50% of the total free amino acids was represented by dipeptides carnosine (average ~130.3 mg/100 g) and anserine (average ~17.9 mg/100 g). Thus, this study for the first time reports the free amino acids profile of water buffalo meat and the content of carnosine and anserine, potentially involved in the darkening meat process and their ratio, that could be used to estimate the water buffalo meat portion in mixed meat products