Effect of Conditioning on the Production of Inulinase by Kluyveromyces marxianus var. bulgaricus through Fed-Batch Fermentation

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Nodule ultrastructure and initial growth of Anadenanthera peregrina (L.) Speg. var. falcata (Benth.) Altschul plants infected with rhizobia

The anatomy and ultrastructure of root nodules of Anadenanthera peregrina var. falcata (Leguminosae-Mimosoideae) were analysed, as was plant growth. To ensure that nodules developed, seedlings were inoculated with a mixture of six strains of rhizobia. Nodules were produced that differed in appearance-and probably also effectiveness-but their structure was similar and they showed characteristics typical of indeterminate nodules, such as persistent meristematic tissue and a gradient of cells at different stages of development. Many starch grains were present in inner cortex cells and interstitial cells of infected tissue. Infected cells were densely packed with bacteroids, which contained many poly-beta-hydroxybutyrate granules. The high incidence of these granules, together with high levels of starch accumulation in interstitial cells, suggested low N-2-fixation efficiency of the rhizobia isolates used for inoculation. In the symbiosomes of early-senescent infected cells, reticulum-like structures, small vesicles and a fibrillar material were observed; these may be related to bacteroid degradation. In the cytoplasm of late-senescent infected cells, many vesicles and membrane-like structures were observed, probably associated with membrane degradation of bacteroids and peribacteroids. The total biomass of plants inoculated with rhizobia was low and their xylopodia and shoots had low levels of N compared with non-inoculated plants fertilized with ammonium nitrate. However, inoculated plants did not show N-deficiency symptoms and grew better than non-inoculated plants without N fertilization. These growth results, together with ultrastructural observations of nodules, suggest that nitrogen fixation of rhizobia isolates associated with Anadenanthera peregrina var. falcata roots is poor. (C) 2002 Annals of Botany Company.

Atividade anti-micobacteriana de extratos de folhas de Byrsonima crassa Nied

In this study, the antimycobacterial activity of chloroformic and methanolic extracts obtained from Byrsonima crassa leaves was evaluated. Antimycobacterial activity was assessed through the microtechnique named Microplate Alamar Blue Assay (MABA) and the promising active principles were identified by spectrophotometric analysis. The chloroformic extract presenting 62.5 μg mL-1 minimum inhibitory concentration (MIC) showed to be active against tuberculosis bacillus. The MIC value of the methanolic extract was 1000 μg mL-1. For the chloroformic one , phytochemical analysis indicated that antimycobacterial activity might be related to the presence of triterpenes.

Biophysical characterization of the recombinant importin-α from neurospora crassa

Neurospora crassa has been widely used as a model organism and contributed to the development of biochemistry and molecular biology by allowing the identification of many metabolic pathways and mechanisms responsible for gene regulation. Nuclear proteins are synthesized in the cytoplasm and need to be translocated to the nucleus to exert their functions which the importin-α receptor has a key role for the classical nuclear import pathway. In an attempt to get structural information of the nuclear transport process in N. crassa, we present herein the cloning, expression, purification and structural studies with N-terminally truncated IMPα from N. crassa (IMPα-Nc). Circular dichroism analysis revealed that the IMPα-Nc obtained is correctly folded and presents a high structural conservation compared to other importins-α. Dynamic light scattering, analytical size-exclusion chromatography experiments and molecular dynamics simulations indicated that the IMPα-Nc unbound to any ligand may present low stability in solution. The IMPα-Nc theoretical model displayed high similarity of its inner concave surface, which binds the cargo proteins containing the nuclear localization sequences, among IMPα from different species. However, the presence of non-conserved amino acids relatively close to the NLS binding region may influence the binding specificity of IMPα-Nc to cargo proteins. Copyright © 2012 Bentham Science Publishers. All Rights Reserved.

Crystallization and preliminary X-ray crystallographic analysis of importin-alpha from Neurospora crassa

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of Byrsonima crassa and Phenolic Constituents on Helicobacter pylori-Induced Neutrophils Oxidative Burst

Byrsonima crassa Niedenzu (Malpighiaceae) is used in Brazilian folk medicine for the treatment of diseases related mainly to gastric ulcers. In a previous study, our group described the gastric protective effect of the methanolic extract from the leaves of B. crassa. The present study was carried out to investigate the effects of methanolic extract and its phenolic compounds on the respiratory burst of neutrophils stimulated by H. pylori using a luminol-based chemiluminescence assay as well as their anti-H. pylori activity. The suppressive activity on oxidative burst of H. pylori-stimulated neutrophils was in the order of methyl gallate > (+)-catechin > methanol extract > quercetin 3-O-alpha-L-arabinopyranoside > quercetin 3-O-beta-D-galactopyranoside > amentoflavone. Methyl gallate, compound that induced the highest suppressive activity with IC50 value of 3.4 mu g/mL, did not show anti-H. pylori activity. B. crassa could be considered as a potential source of natural antioxidant in gastric ulcers by attenuating the effects on the damage to gastric mucosa caused by neutrophil generated reactive oxygen species, even when H. pylori displays its evasion mechanisms.

The alpha and beta subunits of the metalloprotease meprin are expressed in separate layers of human epidermis, revealing different functions in keratinocyte proliferation and differentiation

The zinc endopeptidase meprin (EC 3.4.24.18) is expressed in brush border membranes of intestine and kidney tubules, intestinal leukocytes, and certain cancer cells, suggesting a role in epithelial differentiation and cell migration. Here we show by RT-PCR and immunoblotting that meprin is also expressed in human skin. As visualized by immunohistochemistry, the two meprin subunits are localized in separate cell layers of the human epidermis. Meprin alpha is expressed in the stratum basale, whereas meprin beta is found in cells of the stratum granulosum just beneath the stratum corneum. In hyperproliferative epidermis such as in psoriasis vulgaris, meprin alpha showed a marked shift of expression from the basal to the uppermost layers of the epidermis. The expression patterns suggest distinct functions for the two subunits in skin. This assumption is supported by diverse effects of recombinant meprin alpha and beta on human adult low-calcium high-temperature keratinocytes. Here, beta induced a dramatic change in cell morphology and reduced the cell number, indicating a function in terminal differentiation, whereas meprin alpha did not affect cell viability, and may play a role in basal keratinocyte proliferation.

Benthic foraminifers from the Japan Sea

Five holes were drilled at two sites in the Sea of Japan during Ocean Drilling Program (ODP) Leg 128. Site 798 is located on Oki Ridge at a depth of about 900 m. Sediment age at Site 798 ranges from Pliocene to Holocene. Site 799 is located in the Kita-Yamato Trough at depth of 2000 m and below the present calcite compensation depth (CCD); the sediment ranges from Miocene to Holocene in age. Samples from all holes contain benthic foraminifers. Faunal evidence of downslope displacement is frequent in Holes 799A and 799B. The vertical frequency distribution of some dominant species shows that significant faunal changes occur in Holes 798A-C on Oki Ridge. Based on the faunal change and the thickness of sediments, it appears that the Oki Ridge was uplifted more than 1,000 m during last 4 m.y. Benthic foraminifers also demonstrate that the water depth of Site 799 rapidly changed from upper bathyal to lower bathyal during middle Miocene time. The appearance of benthic foraminifer species common to anaerobic environments suggests that the dysaerobic to anaerobic bottom conditions existed during the evolution of the Sea of Japan. Faunal distributions also suggest that the 'Tertiary-type' species recognized in the Neogene strata of the Japan Sea coastal regions disappeared sequentially from the Sea of Japan during Pliocene to late Pleistocene.

Diatom abundance in ODP Leg 178 holes

This paper documents the biostratigraphic distribution and abundance of diatoms from sites drilled during Ocean Drilling Program Leg 178, off the Pacific margin of the Antarctic Peninsula. Drift sediments cored on the continental rise at Sites 1095, 1096, and 1101 have good recovery and a well-defined paleomagnetic record. Well-preserved diatoms are present throughout the upper Miocene to middle Pliocene and in the upper Quaternary section of these sites. The stratigraphic occurrence of diatom species through these intervals defines numerous datum levels. Diatom events are given absolute age estimates through direct correlation to the established paleomagnetic stratigraphy of Sites 1095, 1096, and 1101. Leg 178 diatom biostratigraphic results enable the development of a regional stratigraphic framework for the Pacific sector of the Southern Ocean and record the interaction of open-ocean and shelf-margin diatom floras.

Pollen and diatom analysis on varved sediments in Lake Jues (Harz Mountains, Germany)

Studies combining sedimentological and biological evidence to reconstruct Holocene climate beyond the major changes, and especially seasonality, are rare in Europe, and are nearly completely absent in Germany. The present study tries to reconstruct changes of seasonality from evidence of annual algal successions within the framework of well-established pollen zonation and 14C-AMS dates from terrestrial plants. Laminated Holocene sediments in Lake Jues (10°20.70' E, 51°39.30' N, 241 m a.s.l.), located at the SW margin of the Harz Mountains, central Germany, were studied for sediment characteristics, pollen, diatoms and coccal green algae. An age model is based on 21 calibrated AMS radiocarbon dates from terrestrial plants. The sedimentary record covers the entire Holocene period. Trophic status and circulation/stagnation patterns of the lake were inferred from algal assemblages, the subannual structure of varves and the physico-chemical properties of the sediment. During the Holocene, mixing conditions alternated between di-, oligo- and meromictic depending on length and variability of spring and fall periods, and the stability of winter and summer weather. The trophic state was controlled by nutrient input, circulation patterns and the temperature-dependent rates of organic production and mineralization. Climate shifts, mainly in phase with those recorded from other European regions, are inferred from changing limnological conditions and terrestrial vegetation. Significant changes occurred at 11,600 cal. yr. BP (Preboreal warming), between 10,600 and 10,100 cal. yr. BP (Boreal cooling), and between 8,400 and 4,550 cal. yr. BP (warm and dry interval of the Atlantic). Since 4,550 cal. yr. BP the climate became gradually cooler, wetter and more oceanic. This trend was interrupted by warmer and dryer phases between 3,440 and 2,850 cal. yr. BP and, likely, between 2,500 and 2,250 cal. yr. BP.

Diatom biostratigraphy, Argon ages and chronology of ODP Hole 183-1138A

A thick Neogene section was recovered in the upper ~300 m of Ocean Drilling Program Hole 1138A, drilled on the Central Kerguelen Plateau in the Indian sector of the Southern Ocean. Sediment lithologies consist primarily of mixed carbonate and biosiliceous clays and oozes, with several thin (1-3 cm) tephra horizons. The tephras are glass rich, well sorted, and dominantly trachytic to rhyolitic in composition. Volcaniclastic material in these horizons is interpreted to have originated from Heard Island, 180 km northwest of Site 1138, and was likely emplaced through both primary ash fall and turbiditic, submarine flows. A Neogene age-depth model for Hole 1138A is constructed primarily from 36 diatom biostratigraphic datums. Nannofossil and planktonic foraminifer biostratigraphy provides supporting age information. Additionally, four high-precision 40Ar-39Ar ages are derived from ash and tephra horizons, and these radiometric ages are in close agreement with the biostratigraphic ages. The integrated age-depth model reveals a reasonably complete lower Miocene to upper Pleistocene section in Hole 1138A, with the exception of a ~1-m.y. hiatus at the Miocene/Pliocene boundary. Another possible hiatus is also identified at the Oligocene/Miocene boundary. High Neogene sedimentation rates and the presence of both calcareous and siliceous microfossils, combined with datable tephra horizons, establish Site 1138 as a suitable target for future drilling legs with paleoceanographic objectives. This report also proposes two new diatom species, Fragilariopsis heardensis and Azpeitia harwoodii, from Pliocene strata of Hole 1138A.

Cenozoic benthic foraminifers from ODP Leg 134 holes

This paper discusses the Paleobathymetric and paleoenvironmental history of the New Hebrides Island Arc and North d'Entrecasteaux Ridge during Cenozoic time based on benthic foraminiferal and sedimentological data. Oligocene and Pliocene to Pleistocene benthic foraminiferal assemblages from Sites 827, 828, 829, and 832 of Ocean Drilling Program (ODP) Leg 134 (Vanuatu) are examined by means of Q-mode factor analysis. The results of this analysis recognize the following bathymetrically significant benthic foraminiferal biofacies: (1) Globocassidulina subglobosa biofacies and Bulimina aculeata-Bolivinita quadrilatera biofacies representing the upper bathyal zone (600-1500 m); (2) Gavelinopsis praegeri-Cibicides wuellerstorfi biofacies, indicating the Pacific Intermediate Water (water depth between 1500 and 2400 m); (3) Tosaia hanzawai-Globocassidulina muloccensis biofacies, Valvulineria gunjii biofacies, and the Melonis barleeanus-Melonis sphaeroides biofacies, which characterize the lower bathyal zone; (4) the Nuttallides umbonifera biofacies, which characterizes the interval between the lysocline (approximately 3500 m) and the carbonate compensation depth (approximately 4500 m); and (5) the Rhabdammina abyssorum biofacies representing the abyssal zone below the carbonate compensation depth. Benthic foraminiferal patterns are used to construct Paleobathymetric and paleogeographic profiles of the New Hebrides Island Arc and North d'Entrecasteaux Ridge for the following age boundaries: late Miocene/Pliocene, early/late Pliocene, Pliocene/Pleistocene, and Pleistocene/Holocene.

Distribution of benthic foraminifera in surface sediments on the continental margin off North-West Africa

The benthic foraminiferal populations along three traverses across the Northwest African continental margin were analyzed on the base of ca. 60 surface sediment samples. Depth ranges of 213 species were established and the main trends of vertical distribution are compared with those known from adjacent regions. Main faunal breaks occure at 100/200 m and 1000/1500 m depth of water. Some species show latitudinal distribution boundaries and the same applies to population density (standing stock), reflecting the regional distribution of nutrients supply by river discharge and upwelling processes. - High proportions of Bolivina test at the lower slope indicate extended downslope transport.