Effects of brain ischemia on intermediate filaments of rat hippocampus

Neurofilaments subunits (NF-H, NF-M, NF-L) and glial fibrillary acidic protein (GFAP) were investigated in the hippocampus of rats after distinct periods of reperfusion (1 to 15 days) following 20 min of transient global forebrain ischemia in the rat. In vitro [¹⁴Ca]leucine incorporation was not altered until 48 h after the ischemic insult, however concentration of intermediate filament subunits significantly decreased in this period. Three days after the insult, leucine incorporation significantly increased while the concentration NF-H, NF-M, and NF-L were still diminished after 15 days of reperfusion. In vitro incorporation of³²P into NF-M and NF-L suffered immediately after ischemia, but returned to control values after two days of reperfusion. GFAP levels decreased immediately after ischemia but quickly recovered and significantly peaked from 7 to 10 days after the insult. These results suggest that transient ischemia followed by reperfusion causes proteolysis of intermediate filaments in the hippocampus, and that proteolysis could be facilitated by diminished phosphorylation levels of NF-M and NF-L.     

Localization of sequences regulating ancestral and acquired sites of esterase6 activity in Drosophila melanogaster

We have broadly defined the DNA regions regulating esterase6 activity in several life stages and tissue types of D. melanogaster using P- element-mediated transformation of constructs that contain the esterase6 coding region and deletions or substitutions in 5' or 3' flanking DNA. Hemolymph is a conserved ancestral site of EST6 activity in Drosophila and the primary sequences regulating its activity lie between -171 and -25 bp relative to the translation initiation site: deletion of these sequences decrease activity approximately 20-fold. Hemolymph activity is also modulated by four other DNA regions, three of which lie 5' and one of which lies 3' of the coding region. Of these, two have positive and two have negative effects, each of approximately twofold. Esterase6 activity is present also in two male reproductive tract tissues; the ejaculatory bulb, which is another ancestral activity site, and the ejaculatory duct, which is a recently acquired site within the melanogaster species subgroup. Activities in these tissues are at least in part independently regulated: activity in the ejaculatory bulb is conferred by sequences between -273 and -172 bp (threefold decrease when deleted), while activity in the ejaculatory duct is conferred by more distal sequences between -844 and -614 bp (fourfold decrease when deleted). The reproductive tract activity is further modulated by two additional DNA regions, one in 5' DNA (-613 to -284 bp; threefold decrease when deleted) and the other in 3' DNA (+1860 to +2731 bp; threefold decrease when deleted) that probably overlaps the adjacent esteraseP gene. Collating these data with previous studies suggests that expression of EST6 in the ancestral sites is mainly regulated by conserved proximal sequences while more variable distal sequences regulate expression in the acquired ejaculatory duct site.      

Phylogenetic diversification of immunoglobulin genes and the antibody repertoire

Immunoglobulins are encoded by a large multigene system that undergoes somatic rearrangement and additional genetic change during the development of immunoglobulin-producing cells. Inducible antibody and antibody-like responses are found in all vertebrates. However, immunoglobulin possessing disulfide-bonded heavy and light chains and domain-type organization has been described only in representatives of the jawed vertebrates. High degrees of nucleotide and predicted amino acid sequence identity are evident when the segmental elements that constitute the immunoglobulin gene loci in phylogenetically divergent vertebrates are compared. However, the organization of gene loci and the manner in which the independent elements recombine (and diversify) vary markedly among different taxa. One striking pattern of gene organization is the "cluster type" that appears to be restricted to the chondrichthyes (cartilaginous fishes) and limits segmental rearrangement to closely linked elements. This type of gene organization is associated with both heavy- and light-chain gene loci. In some cases, the clusters are "joined" or "partially joined" in the germ line, in effect predetermining or partially predetermining, respectively, the encoded specificities (the assumption being that these are expressed) of the individual loci. By relating the sequences of transcribed gene products to their respective germ-line genes, it is evident that, in some cases, joined-type genes are expressed. This raises a question about the existence and/or nature of allelic exclusion in these species. The extensive variation in gene organization found throughout the vertebrate species may relate directly to the role of intersegmental (V<==>D<==>J) distances in the commitment of the individual antibody-producing cell to a particular genetic specificity. Thus, the evolution of this locus, perhaps more so than that of others, may reflect the interrelationships between genetic organization and function.      

Iron(III) binding in DNA solutions: complex formation and catalytic activity in the oxidation of hydrazine derivatives.

A strong interaction between iron(III) and calf thymus DNA at pH 7.4 was demonstrated in the present study by separation of the complex by column chromatography and by the slow kinetics of iron(III) removal from DNA by disodium-1,2-dihydroxybenzene-3,5-disulfonate (Tiron). An equilibrium constant of 2.1 x 10(14) was calculated by measurements of bound iron(III) by flame atomic absorption spectroscopy and assuming a one iron to two nucleotide stoichiometry. Graphic analysis of the interaction however, indicated that DNA has two binding sites for iron(III) characterized by a stoichiometry of one iron to 12 nucleotides and one iron to 2 nucleotides, and association constants of 4.8 x 10(12) and 2.3 x 10(11), respectively. The DNA-iron(III) complex isolated by column chromatography was shown to catalyze the oxidation of both 2-phenylethylhydrazine and methylhydrazine by spin-trapping experiments with alpha-(4-pyridyl 1-oxide)-N-tert-butylnitrone (POBN). By contrast, oxidation of 1,2-dimethylhydrazine was not catalyzed. Catalysis of 2-phenylethylhydrazine oxidation was confirmed by oxygen consumption studies. The results suggest that iron chelated to DNA may be significant in DNA damage induced by oxidizable chemicals.     

PERFUSION FOR MYOCARDIAL REVASCULARIZATION WITHOUT AN ARTIFICIAL OXYGENATOR (New Method to Reduce Surgical Morbidity)

Thirteen patients were submitted to direct myocardial revascularization (saphenous vein graft) without the use of an artificial oxygenator. The perfusion was done by a left ventricle-to-aorta bypass and autogenous oxygenation. Most patients had three grafts implanted plus endarterectomy of the distal right coronary artery. There was one hospital death that was apparently not related to the method used. Perfusion time ranged from 45 minutes to 4 hours. Body temperature during perfusion was kept between 25 and 30 degrees C. Perfusion flow was maintained between 25 to 50 ml per kg of body weight per minute. Ischemic, hypothermic cardiac arrest was employed. We demonstrated for the first time that perfusion for this kind of heart surgery could be done with no artificial oxygenators and, apparently, is safer for the patients. There were no bleeding problems even in perfusions as long as 4 hours. There was no respiratory dysfunction, and artificial respiration was used for only 6 to 12 hours. The patients awoke at the end of surgery with no signs or symptoms of central nervous system damage, and vasopressor drugs were rarely used after surgery. Although the experience is very small, it suggests that many postoperative problems, especially those related to bleeding and respiratory dysfunction may be reduced or eliminated by this new method.     

An ethological analysis of types of agonistic interaction in a captive group of Java-monkeys (Macaca fascicularis)

The primate literature provides many indications not only that the nature of dyadic interactions is to a large extent determined by the relations of the interacting animals with others and between these others, but also of the existence of polyadic interactions in which more than two individuals are simultaneously involved. The objectives of the present study are to obtain a quantitative categorization of the agonistic interaction types of captive Java-monkeys and an analysis of their dynamics. After having described the agonistic behaviour patterns of Java-monkeys we shall discuss the categorization of agonistic interaction types (depending on the number of involvees: “dyads”, “triads” and “polyads”), the way in which these types can be further differentiated on the basis of the nature and the direction of the behaviours shown (e.g., different types of alliances), and the existence of so-called “sub-directed” behaviours (i.e., non-agonistic behaviours which are shown towards a dominant third animal more or less simultaneously with aggressive behaviour directed towards an opponent). The analysis indicates that agonistic behaviour is different both in its form and its regulation in interactions of different complexity. This research was supported in part by a government grant (i.e.: Beleidsruimte project: 16-21-06, “Brain and Behaviour”) to the first author. The investigation was supported by a grant from the Beleidsruimtemiddelen Hersenen en Gedrag to the first author.     

Yellow‐chevroned Parakeet (Brotogeris chiriri) abundance and canopy foraging at a dry forest in western Brazil

I studied Brotogeris chiriri abundance and foraging activity at a dry forest of the Urucum mountains in western Mato Grosso do Sul, Brazil, to evaluate their relationships with food resource production. Brotogeris chiriri abundance sharply increased during the early wet season (mainly October 2001) when it mostly foraged for fleshy fruits. At that time Protium heptaphyllum, one of the most common tree species, bore a large crop of fruits, the arils of which were extensively consumed by B. chiriri. Conversely, only a few parakeets were recorded foraging from the late wet to the late dry season, when dry fruit production predominated. The monthly pattern of parakeet abundance paralleled both its monthly pattern of foraging activity and fleshy fruit availability. Moreover, the variations in foraging activity were highly correlated to fleshy fruit production. Thus, data presented here evidenced the effect of both fruiting pulses and a common tree species that produced a large and ephemeral fruit crop, on the dynamic of a small and mobile canopy forager at a primary dry forest.     

The role of snail aestivation in transmission of schistosomiasis in changing climatic conditions

Schistosomiasis vector snails are subjected to extreme seasonal changes, particularly in ephemeral rivers and lentic waterbodies. In the tropics, aestivation is one of the adaptive strategies for survival and is used by snails in times of extremely high temperatures and desiccation. Aestivation therefore plays an important role in maintaining the transmission of schistosomiasis. This review assesses the possible impacts of climate change on the temporal and spatial distribution of schistosomiasis-transmitting snails with special emphasis on aestivation, and discusses the effect of schistosome infection on aestivation ability. The impacts of parasite development on snails, as well as physiological changes, are discussed with reference to schistosomiasis transmission. This review shows that schistosome-infected snails have lower survival rates during aestivation, and that those that survive manage to get rid of the infection. In general, snail aestivation ability is poor and survival chances diminish with time. Longer dry periods result in fewer, as well as uninfected, snails. However, the ability of the surviving snails to repopulate the habitats is high.     

Glucose Metabolism Disorder Is Associated with Pulmonary Tuberculosis in Individuals with Respiratory Symptoms from Brazil

BackgroundDiabetes mellitus (DM) has been associated with increased risk for pulmonary tuberculosis (PTB) in endemic settings but it is unknown whether PTB risk is also increased by pre-DM. Here, we prospectively examined the association between glucose metabolism disorder (GMD) and PTB in patients with respiratory symptoms at a tuberculosis primary care reference center in Brazil.MethodsOral glucose tolerance test was performed and levels of fasting plasma glucose and glycohemoglobin (HbA1c) were measured in a cohort of 892 individuals presenting with respiratory symptoms of more than two weeks duration. Patients were also tested for PTB with sputum cultures. Prevalence of pre-DM and DM (based on HbA1c) was estimated and tested for association with incident PTB. Other TB risk factors including smoking history were analyzed.ResultsThe majority of the study population (63.1%) exhibited GMD based on HbA1c ≥5.7%. Patients with GMD had higher prevalence of PTB compared to normoglycemic patients. Individuals with DM exhibited increased frequency of TB-related symptoms and detection of acid-fast bacilli in sputum smears. Among patients with previous DM diagnosis, sustained hyperglycemia (HbA1c ≥7.0%) was associated with increased TB prevalence. Smoking history alone was not significantly associated with TB in our study population but the combination of smoking and HbA1c ≥7.0% was associated with 6 times higher odds for PTB.ConclusionsSustained hyperglycemia and pre-DM are independently associated with active PTB. This evidence raises the question whether improving glycemic control in diabetic TB patients would reduce the risk of TB transmission and simultaneously reduce the clinical burden of disease. A better understanding of mechanisms underlying these associations, especially those suggesting that pre-DM may be a factor driving susceptibility to TB is warranted.