Age-Related Changes in Ang II Receptor Localization and Expression in the Developing Auditory Pathway.

We studied Ang II receptor localization in different nuclei of the auditory system, by means of binding autoradiography, during brain development. The inferior colliculus (IC), a large midbrain structure which serves as an obligatory synaptic station in both the ascending and descending auditory pathways, exhibited high Ang II AT2 binding at all ages (P0, P8, P15, P30), being maximal at P15. These observations were confirmed by in situ hybridization and immunofluorescence at P15, demonstrating that AT2 receptor mRNA localized at the same area recognized by AT2 antibodies and anti ß III-tubulin suggesting the neuronal nature of the reactive cells. Ang II AT1 receptors were absent at early developmental ages (P0) in all nuclei of the auditory system and a low level was observed in the IC at the age P8. AT2 receptors were present at ventral cochlear nucleus and superior olivary complex, being higher at P15 and P8, respectively. We also explored the effect of prenatal administration of Ang II or PD123319 (AT2 antagonist) on binding of Ang II receptors at P0, P8, P15. Both treatments increased significantly the level of AT2 receptors at P0 and P8 in the IC. Although total binding in the whole IC from P15 animals showed no difference between treatments, the central nucleus of the IC exhibited higher binding. Our results supports a correlation between the timing of the higher expression of Ang II AT2 receptors in different nuclei, the onset of audition and the establishment of neuronal circuits of the auditory pathway.

Wing geometric morphometrics and molecular assessment of members in the Albitarsis Complex from Colombia.

Malaria parasites are transmitted to humans by female mosquitoes of the genus Anopheles. The Albitarsis Complex harbours at least eight species not readily differentiable by morphology. This complicates the determination of those species involved in malaria transmission and the implementation of targeted and effective vector control strategies. In Colombia, there is little information about the identity and distribution of the Albitarsis Complex members. In this work, COI DNA barcoding was used to assign specimens Anopheles albitarsis s.l. to any of the previously designated species of the Albitarsis Complex. Two molecular operational taxonomic units (MOTUs), differentially distributed in Colombia, were detected, A. albitarsis I in the NW and NE, and A. albitarsis F, E and NE Colombia. In contrast, nuclear white gene and ITS2 sequence analyses did not allow differentiating between the MOTUs. Wing landmark-based geometric morphometrics applied to explore intertaxa phenotypic heterogeneity showed a subtle but significant difference in size, while shape did not allow the separation of the MOTUs. In general, the multiple marker analysis was not supportive of the existence in Colombia of more than one species of the Albitarsis Complex.

Anammox bacteria enrichment and characterization from municipal activated sludge.

A sustainable option for nitrogen removal is the anaerobic ammonium-oxidizing (anammox) process in which ammonium is oxidized to nitrogen gas with nitrite as electron acceptor. Application of this process, however, is limited by the availability of anammox biomass. In this study, two Brocadia-like anammox phylotypes were successfully enriched, detected and identified from an activated sludge taken from a domestic wastewater treatment plant (Minas Gerais, Brazil) employing a Sequencing Batch Reactor (SBR). The dominant phylotype was closely related to 'Candidatus Brocadia sinica', but one clone seemed to represent a novel species for which we propose the name 'Candidatus Brocadia brasiliensis'. Based on Fluorescence in situ hybridization (FISH) analysis, this enrichment led to a relative population size of 52.7% (±15.6) anammox bacteria after 6 months of cultivation. The cultivation process can be divided into three phases: phase 1 (approximately 25 days) was characterized by heterotrophic denitrification metabolism, phase 2 was the propagation phase and phase 3 (from the 87th day onwards), in which significant anammox activity was detected. A long-term performance of the SBR showed a near perfect removal of nitrite based on the influent NO(2)(-)-N concentration of 61-95 mg L(-1). The average ammonia removal efficiency was 90% with the influent NH(4)(+)-N concentration of 55-82 mg L(-1). Therefore, anammox cultivation and enrichment from activated sludge was possible under a controlled environment within 3 months.

Do leaf-litter attributes affect the richness of leaf-litter ants?

The search for factors shaping leaf-litter ant communities has received particular attention due to the essential role of these insects in many ecological processes. Here, we aimed to investigate how the number of leaves and leaf morphotypes affect the litter-ant species density at forest edge and interior in an Atlantic Forest remnant in the state of Alagoas, Brazil. This study was developed based on 28 litter plots (1m² each), 14 in the forest interior and 14 in the forest edge. As we early expected, ant species density increased with increasing both the number of leaves and the number of leaf morphotypes, but this result was clearly influenced by plot location. Contrasting with the forest interior, ant species density did not increase as the number of leaves increased in the forest edge. Possibly, factors such as plant species richness, vegetation structure and environmental conditions affect ant species density as well as promote a patchy distribution of species in ant communities along the edge-to-interior gradient. Our findings suggest that edge-affected forests present more simplified ant communities, with different factors shaping its structure. We encourage future studies to include leaf litter heterogeneity as one of the explanatory variables investigated.

Investigation of aerobic and anaerobic ammonium-oxidising bacteria presence in a small full-scale wastewater treatment system comprised by UASB reactor and three polishing ponds.

This work applied PCR amplification method and Fluorescence in situ hybridisation (FISH) with primers and probes specific for the anammox organisms and aerobic ammonia-oxidising beta-Proteobacteria in order to detect these groups in different samples from a wastewater treatment system comprised by UASB reactor and three polishing (maturation) ponds in series. Seven primer pairs were used in order to detect Anammox bacteria. Positive results were obtained with three of them, suggesting that Anammox could be present in polishing pond sediments. However, Anammox bacteria were not detected by FISH, indicating that they were not present in sediment samples, or they could be present but below FISH detection limit. Aerobic ammonia- and nitrite-oxidising bacteria were verified in water column samples through Most Probable Number (MPN) analysis, but they were not detected in sediment samples by FISH. Ammonia removal efficiencies occurred systematically along the ponds (24, 32, and 34% for polishing pond 1, 2, and 3, respectively) but the major reaction responsible for this removal is still unclear. Some nitrification might have occurred in water samples because some nitrifying bacteria were present. Also Anammox reaction might have occurred because Anammox genes were detected in the sediments, but probably this reaction was too low to be noticed. It is important also to consider that some of the ammonia removal observed might be related to NH(3) stripping, associated with the pH increase resulting from the intensive photosynthetic activity in the ponds (mechanism under investigation). Therefore, it can be concluded that more than one mechanism (or reaction) might be involved in the ammonia removal in the polishing ponds investigated in this study.

Diagnosis of paracoccidioidomycosis by a dot blot assay using a recombinant Paracoccidioides brasiliensis p27 protein.

A variety of immunological methods have proven useful for Paracoccidioidomycosis (PCM) diagnosis; however, they are often time consuming and many lack sensitivity and specificity, partially attributed to the use of crude antigens, which give cross reactivity. Until now, attempts to clone and express Paracoccidioides brasiliensis immunodominant antigens have presented difficulties of process and problems of cost. In an attempt to obtain a more rapid, sensitive, and specific test for PCM diagnosis, we subcloned the P. brasiliensis p27 gene and used the recombinant protein as the antigen in dot blot assays to evaluate its usefulness in paracoccidioidomicosis diagnosis. The development of an optimised procedure for p27 recombinant protein purification and production led to an easier and less expensive process than the one previously used in our laboratory and allowed the availability of enough purified protein for its evaluation as the antigen in the dot blot assays. In these assays, antibodies present in ten serum samples from seven patients with PCM recognised the recombinant protein showing a sensitivity of 100% with a specificity of 98%. These results confirm the value of the 27-kDa recombinant antigen in the serodiagnosis of paracoccidioidomycosis and that the dot blot format is an alternative to the immunoenzymatic assay procedure.

Needling versus medical treatment in encapsulated blebs. A randomized, prospective study.

OBJECTIVE: The purpose of the study is to compare the efficacy and safety of transconjunctival needling and medical treatment in eyes with encapsulated blebs. DESIGN: A randomized, prospective study. PARTICIPANTS: Two hundred eighty-two eyes that underwent a guarded filtration procedure between January 1994 and January 1996 at the Glaucoma Service of University of Campinas. INTERVENTION: Encapsulated blebs developed in 25 (8.9%) of 282 eyes and were randomized to either needling (n = 14) or medical treatment with aqueous humor suppressants (n = 11). If one treatment failed to maintain intraocular pressures (IOPs) below 20 mmHg, the other treatment was initiated. If both methods failed, surgical revision or further glaucoma surgery was performed. Complete success was defined as IOP less than 20 mmHg after one treatment method. Qualified success was defined when IOPs less than 20 mmHg were obtained with both treatment methods, whereas failure was defined when IOP greater than 20 mmHg or when further surgery was indicated. MAIN OUTCOME MEASURES: Intraocular pressure, vision, and number of antiglaucoma medications. RESULTS: After a mean follow-up of 9.6 months, medical treatment alone was successful in ten patients (90.9%), whereas needling alone was successful in one patient (7.1%) (P = 0.00003). In the needling group, 92.9% of the eyes required aqueous humor suppressants, and 57.1% were considered qualified successes at the last follow-up (mean = 10.1 months). At the last follow-up examination, there was no statistically significant difference between the mean number of medications in both groups (P = 0.797). Further glaucoma surgery was performed in five patients (35.7%) undergoing needling and one patient (9.1%) receiving medical treatment (P = 0.162). CONCLUSIONS: Medical treatment with digital pressure should be used as the initial treatment in eyes with encapsulated blebs. Needling procedures or surgical revision, methods that are more invasive and potentially associated with severe complications, should be limited to the small percentage of eyes that do not respond to medical treatment.

Biochemical and histopathological alterations induced in rats by Tityus serrulatus scorpion venom and its major neurotoxin tityustoxin-I.

Intravenous injection into the rat of sublethal doses of Tityus serrulatus scorpion venom (100 micrograms protein/kg) or its major neurotoxin tityustoxin-I (TsTX-I, 20 micrograms/kg) caused, 30-180 min after injection, statistically significant increases in the serum levels of aspartate aminotransferase, amylase, creatine kinase and lactate dehydrogenase, as well as hyperglycemia, a high level of plasma free fatty acids and a low level of liver glycogen. The in vitro serum levels of the above enzymes did not change. For alanine aminotransferase, gamma-glutamyl transferase and alkaline phosphatase, neither in vitro nor in vivo alterations were observed. The whole venom and TsTX-I caused hepatic congestion with hemolysis and hydropic degeneration. Other histological lesions included edema and congestion with subpleural hemorrhage in the lungs, hypertrophy of fibers with degeneration areas in the heart, and congestion and hemorrhage in the kidneys. In the salivary glands, alterations to the acini and ductules were visible. In the adrenal glands no morphological alterations could be detected at the studied doses. The results suggest that the in vivo enzymatic and histopathological alterations are due to tissue lesions evoked by the whole venom and TsTX-I. An indirect effect, however, induced by stimulation of acetylcholine and catecholamine release in the postganglionic nerve terminals, cannot be excluded.

Fractionation of Bothrops pirajai snake venom: isolation and characterization of piratoxin-I, a new myotoxic protein.

Whole desiccated venom of Bothrops pirajai was fractionated on a gel filtration (Sephadex G-75) column. Phospholipase A2, arginine esterase and clotting activity profiles of the six fractions (SI to SVI) obtained were determined. Fraction SIV from the gel filtration column was subjected to chromatography on SP-Sephadex C-25. It was resolved into five subfractions (SIV-SP1, to SIV-SP5). Fractions SIV-SP1, SIV-SP2 and SIV-SP3 showed phospholipase A2 activity but, among these fractions, only SIV-SP3 was homogeneous. Induction of myonecrosis by SIV-SP3, SIV-SP4 and SIV-SP5 was demonstrated by their ability to release serum creatine kinase, and for SIV-SP5, to induce histological alterations in the injected mouse muscle. Chemical characterization by determination of mol. wts, isoelectric focusing and direct manual sequencing of the N-terminal region was performed for SIV-SP3, SIV-SP4 and SIV-SP5. When compared with bothropstoxin-I, the myotoxin SIV-SP5 showed the same total number of amino acid residues (121) and constant molar ratio for all but three amino acids. We have named this toxin piratoxin-I (PrTX-I).

A new serotype of Bacillus thuringiensis from Colombia toxic to mosquito larvae.

During a survey conducted in Colombia a new isolate of Bacillus thuringiensis that showed toxicity toward Culex quinquefasciatus, Cx. pipiens, Aedes aegypti, and Anopheles stephensi larvae was isolated. Parasporal crystals were spherical in shape and showed a great degree of similarity with those produced by the reference strain of Bacillus thuringiensis subsp. israelensis. Supernatant fraction of the whole culture was not toxic, and heat-stable exotoxin production was negative. Catalase, urease, arginine dihydrolase, amylase, lecithinase, acetyl-methyl-carbinol, and gelatinase production were positive. Hemolysis on sheep blood agar was alpha-type. The isolate 163-131 showed natural resistance to azolocillin and was sensible to cephoperazone, cephalotin, nalidixic acid, and trimetoprin sulfametoxazole. Flagellar agglutination showed a specific H 30 antigen which allows individualization of this strain as a new serotype and the subspecies name of medellin is proposed.