Extracellular α-amylase from Thermus filiformis Ork A2: purification and biochemical characterization

An extracellular α-amylase produced by the thermophilic bacterium Thermus filiformis Ork A2 was purified from cell-free culture supernatant by ion exchange chromatography. The molecular mass was estimated to be 60 000 Da by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme was rich in both basic and hydrophobic amino acids, presenting the following NH₂-terminal amino acid sequence: Thr-Ala-Asp-Leu-Ile-Val-Lys-Ile-Asn-Phe. Amylolytic activity on soluble starch was optimal at pH 5.5–6.0 and 95°C, and the enzyme was stable in the pH range of 4.0–8.0. Calcium enhanced thermostability at temperatures above 80°C, increasing the half-life of activity to more than 8 h at 85°C, 80 min at 90°C, and 19 min at 95°C. Ethylenediaminetetraacetic acid (EDTA) inhibited amylase activity, the inhibition being reversed by the addition of calcium or strontium ions. The α-amylase was also inhibited by copper and mercuric ions, and p-chloromercuribenzoic acid, the latter being reversed in the presence of dithiothreitol. Dithiothreitol and β-mercaptoethanol activated the enzyme. The α-amylase exhibited Michaelis-Menten kinetics for starch, with a K _m of 5.0 mg·ml⁻¹ and k _cat/K _m of 5.2 × 10⁵ ml·mg⁻¹ s⁻¹. Similar values were obtained for amylose, amylopectin, and glycogen. The hydrolysis pattern was similar for maltooligosaccharides and polysaccharides, with maltose being the major hydrolysis product. Glucose and maltotriose were generated as secondary products, although glucose was produced in high levels after a 6-h digestion. To our knowledge this is the first report of the characterization of an α-amylase from a strain of the genus Thermus. Received: June 2, 1997 / Accepted: September 16, 1997     

Effect of the oxygen supply on pattern of growth and corrinoid and organic acid production of Propionibacterium shermanii

Propionibacterium shermanii CDB 10014 is able to grow even at high oxygen transfer rates (24.0 mmol O₂ l⁻¹ h⁻¹), in contrast to reports in the specialised literature, where all Propionibacteria are considered oxygen-sensitive microorganisms. Propionic acid is the main product in anaerobiosis. The presence of oxygen in the system leads to an inhibition of propionic acid production while acetic acid formation is enhanced. At high oxygen supply rates no propionic acid is produced and acetic acid is the main product. Lactic acid is also produced in reasonable quantities (2.7 g l⁻¹). The growth rate (μ_max) is higher in anaerobiosis (0.19 h⁻¹) than in aerobiosis (0.12–0.15 h⁻¹). The cell yield is higher in aerobiosis (0.18–0.22 g g⁻¹) than in anaerobiosis (0.14 g g⁻¹) suggesting the oxidative metabolism of glucose by Propionibacterium shermanii CDB 10014. No corrinoid production was detected at oxygen transfer rates of more than 13.6 mmol l⁻¹ h⁻¹. Received: 10 September 1997 / Received revision: 6 January 1998 / Accepted: 9 January 1998     

The response of an Eastern Amazonian rain forest to drought stress: results and modelling analyses from a throughfall exclusion experiment

Warmer and drier climates over Eastern Amazonia have been predicted as a component of climate change during the next 50–100 years. It remains unclear what effect such changes will have on forest–atmosphere exchange of carbon dioxide (CO₂) and water, but the cumulative effect is anticipated to produce climatic feedback at both regional and global scales. To allow more detailed study of forest responses to soil drying, a simulated soil drought or 'throughfall exclusion' (TFE) experiment was established at a rain forest site in Eastern Amazonia, Brazil, for which time-series sap flow and soil moisture data were obtained. The experiment excluded 50% of the throughfall from the soil. Sap flow data from the forest plot experiencing normal rainfall showed no limitation of transpiration throughout the two monitored dry seasons. Conversely, data from the TFE showed large dry season declines in transpiration, with tree water use restricted to 20% of that in the control plot at the peak of both dry seasons. The results were examined to evaluate the paradigm that the restriction on transpiration in the dry season was caused by limitation of soil-to-root water transport, driven by low soil water potential and high soil-to-root hydraulic resistance. This paradigm, embedded in the soil–plant–atmosphere (SPA) model and driven using on-site measurements, provided a good explanation ( R ² > 0.69) of the magnitude and timing of changes in sap flow and soil moisture. This model-data correspondence represents a substantial improvement compared with other ecosystem models of drought stress tested in Amazonia. Inclusion of deeper rooting should lead to lower sensitivity to drought than the majority of existing models. Modelled annual GPP declined by 13–14% in response to the treatment, compared with estimated declines in transpiration of 30–40%.     

Relief influence on the spatial distribution of the Atlantic Forest cover on the Ibiúna Plateau, SP.

Several studies suggest that, on a large scale, relief conditions influence the Atlantic Forest cover. The aim of this work was to explore these relationships on a local scale, in Caucaia do Alto, on the Ibiúna Plateau. Within an area of about 78 km(2), the distribution of forest cover, divided into two successional stages, was associated with relief attribute data (slope, slope orientation and altitude). The mapping of the vegetation was based on the interpretation of stereoscopic pairs of aerial photographs, from April 2000, on a scale of 1:10,000, while the relief attributes were obtained by geoprocessing from digitalized topographic maps on a scale of 1:10,000. Statistical analyses, based on qui-square tests, revealed that there was a more extensive forest cover, irrespective of the successional stage, in steeper areas (>10 degrees) located at higher altitudes (>923 m), but no influence of the slope orientation. There was no sign of direct influence of relief on the forest cover through environmental gradients that might have contributed to the forest regeneration. Likewise, there was no evidence that these results could have been influenced by the distance from roads or urban areas or with respect to permanent preservation areas. Relief seems to influence the forest cover indirectly, since agricultural land use is preferably made in flatter and lower areas. These results suggest a general distribution pattern of the forest remnants, independent of the scale of study, on which relief indirectly has a strong influence, since it determines human occupation.     

Expression of Parkin,APC, APE1, and Bcl-xL in Colorectal Polyps

Colorectal cancer can develop through molecular, chromosomal, and epigenetic cumulative changes that transform the normal intestinal epithelium into the colorectal polyps, called conventional adenomas (CAs) or serrated polyps (SPs), recognized as precursors of invasive colorectal neoplasia. These benign lesions need to explore the morphology, histological diagnosis, and biomarkers profile to accurately characterize lesions with potential for evolution to cancer. This study aimed to correlate the immunohistochemical expression of Parkin and Adenomatous Polyposis Coli (APC; tumor suppressors), Human Apurinic/Apyrimidinic endonuclease 1 (APE1), and B-cell lymphoma-extra-large (Bcl-xL; oncogenic proteins) in sporadic colorectal polyps with clinical, endoscopic, and diagnostic data. Immunohistochemical analysis was performed on tissue microarray samples of 306 polyps. Based on the Allred score, the expressions were graduated in the cytoplasm and nucleus of superficial and cryptic cells. There was higher Parkin nuclear expression (p=0.006 and 0.010) and APC cytoplasmic expression in cryptic cells (p<0.001) in SPs. CAs, APE1 (p<0.001) and Bcl-xL (p<0.001) were more expressed in the nuclei and cytoplasms, respectively. These results are related to the biological role proposed for these proteins in cellular functions. They can contribute to the diagnosis criteria for polyps and improve the knowledge of biomarkers that could predict cancer development:     

Length–weight relationships for fishes caught by shrimp trawl in Santa Catarina coast,south Atlantic,Brazil

Length–weight relationships (TW = a TL^b) were estimated for fish species caught by bottom shrimp trawl north of Santa Catarina Island (Brazil) from October 2003 to September 2004. For three of the species this is the first LWR data in Brazil.     

Selection of Iberian barbel Barbus bocagei (Steindachner, 1864) for orifices and notches upon different hydraulic configurations in an experimental pool-type fishway

Trends in fish passage studies are increasingly moving towards a holistic approach in considering the movements of a wide range of fish species and other aquatic fauna. In this context, it is important to consider not only salmonids and other game species, but also coarse species such as cyprinids. Moreover, knowledge of their behaviour when confronted with fishways is limited, particularly their upstream movements with regard to different hydraulic configurations. To address this deficit, research was conducted in an indoor full-scale pool-type fishway to assess the cyprinid Iberian barbel Barbus bocagei (Steindachner, 1864) response to the simultaneous presence of submerged orifices and surface notches with adjustable dimensions in association with two different flow regimes over the notches, i.e. plunging and streaming. Overall, orifices (76%) were the clear choice to negotiate the fishway. When the flow was in the plunging mode, a significantly higher proportion of fish selected the orifices to move upstream, but the proportions were equal during the streaming mode. The time taken to enter the fishway differed significantly according to the chosen discharge device and was lower for submerged orifices (52 s to 35 min.). The present study identified key factors in Iberian barbel upstream movements, which provide direct applications to future fishway designs for this species.     

A rearranged chamigrene derivative and its potential biogenetic precursor from a new species of the marine red algal genus Laurencia (rhodomelaceae)

The structures of a novel rearranged sesquiterpenoid and a biogenetically-related chamigrene derivative have been determined by combined spectral and chemical methods. These sesquiterpenoids were components of an undescribed Laurencia species, and each was toxic toward the damselfish Pomacentrus coeruleus.