Search

For insects that depend on one or more bacterial endosymbionts for survival, it is critical that these bacteria are faithfully transmitted between insect generations. Cicadas harbor two essential bacterial endosymbionts, " Candidatus Sulcia muelleri" and " Candidatus Hodgkinia cicadicola." In some cicada species, Hodgkinia has fragmented into multiple distinct but interdependent cellular and genomic lineages that can differ in abundance by more than two orders of magnitude. This complexity presents a potential problem for the host cicada, because low-abundance but essential Hodgkinia lineages risk being lost during the symbiont transmission bottleneck from mother to egg. Here we show that all cicada eggs seem to receive the full complement of Hodgkinia lineages, and that in cicadas with more complex Hodgkinia this outcome is achieved by increasing the number of Hodgkinia cells transmitted by up to 6-fold. We further show that cicada species with varying Hodgkinia complexity do not visibly alter their transmission mechanism at the resolution of cell biological structures. Together these data suggest that a major cicada adaptation to changes in endosymbiont complexity is an increase in the number of Hodgkinia cells transmitted to each egg. We hypothesize that the requirement to increase the symbiont titer is one of the costs associated with Hodgkinia fragmentation. IMPORTANCE Sap-feeding insects critically rely on one or more bacteria or fungi to provide essential nutrients that are not available at sufficient levels in their diets. These microbes are passed between insect generations when the mother places a small packet of microbes into each of her eggs before it is laid. We have previously described an unusual lineage fragmentation process in a nutritional endosymbiotic bacterium of cicadas called Hodgkinia In some cicadas, a single Hodgkinia lineage has split into numerous related lineages, each performing a subset of original function and therefore each required for normal host function. Here we test how this splitting process affects symbiont transmission to eggs. We find that cicadas dramatically increase the titer of Hodgkinia cells passed to each egg in response to lineage fragmentation, and we hypothesize that this increase in bacterial cell count is one of the major costs associated with endosymbiont fragmentation., (Copyright © 2018 Campbell et al.)

Two strategies have been described for cold tolerance in arthropods: (1) freeze-tolerant organisms, which can survive the formation of ice crystals and (2) freeze-avoidant organisms, which prevent the ice crystal formation by super cooling their internal fluids. We studied two arthropods from the Andean Range in central Chile (2400 m a.s.l.), the stick insect Agathemera crassa commonly named as "Chinchemolle", and the tarantula spider Euathlus condorito commonly named as "Araña pollito", in order to evaluate how they respond to low temperatures at the physiological and molecular levels. We sampled the soil temperature during one year to track the temperature changes that these organisms must overcome. We found minimum temperatures around -6 °C in autumn, while the temperature were stable at 0 °C in winter due to the snow. The average field-cooling rate was 0.01 ± 0.006 °C min -1 . For both arthropods we determined the super cooling point (SCP) at a cooling rate of 1 °C min -1 and its subsequent survival, finding that A. crassa is a freezing tolerant organism with a SCP of -3.8 ± 1.8 °C and 100% survival, while E. condorito is a freezing avoidant organism with a SCP of -3.0 ± 1.3 °C and 0% survival. The SCP and survival were not affected by the season in which individuals were collected, the SCP was significantly affected by the cooling rate of the experiment. Both species had low molecular weight cryoprotective in their hemolymph that could explain their cold-tolerance behavior. Glucose, glycerol, and trehalose were found in A. crassa's hemolymph, only glucose and glycerol were found in E. condorito's. We analyzed the hemolymph proteins and found no seasonal differences in composition for either species and also we detected protein antifreeze activity in the hemolymph from both arthropods., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Here in, we report the preparation and evaluation of four 3-hydroxy-piperidine-N-benzyl-aryl-acylhydrazone derivatives (6a-d) for their potential antinociceptive activity. In the tail flick test, compounds 6a and 6d exhibited a significant increase in the latency time of the animals, in comparison to the control group. These two compounds also showed a significant increase in the nociceptive threshold from 1 to 6 h after treatment in the CCI neuropathic pain model. In both cases, the antinociceptive activity was blocked by naloxone, suggesting an opioid mechanism of action, but without sedative or motor coordination effects., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Bacterial endosymbionts that provide nutrients to hosts often have genomes that are extremely stable in structure and gene content. In contrast, the genome of the endosymbiont Hodgkinia cicadicola has fractured into multiple distinct lineages in some species of the cicada genus Tettigades To better understand the frequency, timing, and outcomes of Hodgkinia lineage splitting throughout this cicada genus, we sampled cicadas over three field seasons in Chile and performed genomics and microscopy on representative samples. We found that a single ancestral Hodgkinia lineage has split at least six independent times in Tettigades over the last 4 million years, resulting in complexes of between two and six distinct Hodgkinia lineages per host. Individual genomes in these symbiotic complexes differ dramatically in relative abundance, genome size, organization, and gene content. Each Hodgkinia lineage retains a small set of core genes involved in genetic information processing, but the high level of gene loss experienced by all genomes suggests that extensive sharing of gene products among symbiont cells must occur. In total, Hodgkinia complexes that consist of multiple lineages encode nearly complete sets of genes present on the ancestral single lineage and presumably perform the same functions as symbionts that have not undergone splitting. However, differences in the timing of the splits, along with dissimilar gene loss patterns on the resulting genomes, have led to very different outcomes of lineage splitting in extant cicadas., Competing Interests: The authors declare no conflict of interest.

Objective: Identifying the types of non-psychotic mental disorders in adult women who suffered intimate partner violence in the literature., Method: An integrative review carried out in the MEDLINE, CINAHL, LILACS, Web of Science and SCOPUS databases., Results: We selected 19 articles published in international journals in English, with a predominance of cross-sectional study studies (78.9%). The most common types of non-psychotic mental disorders were: depression (73.7%) and post-traumatic stress disorder (52.6%). It was observed that 78.9% of the articles presented a 2C level of evidence., Conclusion: Studies have shown that adult women who are victims of intimate partner violence mostly suffer from depression and post-traumatic stress disorder, as well as other morbidities; a fact that highlights how devastating violence by an intimate partner can impact on the mental health of those who experience it.

In this article, we assessed the effect of the rearing temperature on life history traits of the poorly known fly Phasmovora phasmophagae (Diptera: Tachinidae), a parasitoid of Agathemera crassa (Phasmatodea: Agathemeridae) in order to: i) test the effect of ambient temperature on life history traits and ii) assess the potential trade-off between reproduction and survival. Parasitoids were obtained from a population of hosts located in the Andes range of central Chile. Upon emergence from the host parasitoids were randomly allocated to three thermal treatments (15°C, 22.5°C and 30°C) and several life history traits were measured. We recorded higher survival at 15°C and 22.5°C and a lower survival at 30°C.We found differences for both body mass and head width among thermal treatments. In females, body mass was higher at 15°C than at 30°C. An effect of breeding temperature and sex was observed only for developmental time. In addition, males reared at different temperatures during the pupal stage and held as adults at 22.5°C, exhibited no differences in longevity between treatments. A significant effect of temperature on the mass of ovaries and lipid was recorded in females. These patterns suggest a trade-off between reproduction and survival. Overall, data seem to support the "colder is better" hypothesis, because Andean parasitoid P. phasmophagae inhabiting and experimentally reared in colder environments have a higher performance in all environments., (Copyright © 2017. Published by Elsevier Ltd.)

Objective: To analyze the self-inflicted violence by exogenous poisoning reported in a health service., Methods: Epidemiological, retrospective and analytical study in an emergency care in the city of Teresina, Piauí. The study took place in January and February of 2015, upon review of all cases of self-inflicted violence by exogenous poisoning reported to the Information System on Diseases of Compulsory Declaration, from 2009 to 2014. The Pearson chi-square test was used for analysis., Results: 277 victims of self-inflicted violence by exogenous poisoning were reported, with 10.5% having died by suicide. There was an association between death and the age, education, area of occurrence and type of exposure, as well as between the type of exposure and the amount of agents used., Conclusion: The results help to define prevention strategies considering vulnerable groups and the complexity of the factors associated with self-inflicted violence.

The adoption of carbon inventories for dairy farms in tropical countries based on models developed from animals and diets of temperate climates is questionable. Thus, the objectives of this study were to estimate enteric methane (CH4) emissions through the SF6 tracer gas technique and through equations proposed by the Intergovernmental Panel on Climate Change (IPCC) Tier 2 and to calculate the inventory of greenhouse gas (GHG) emissions from two dairy systems. In addition, the carbon balance of these properties was estimated using enteric CH4 emissions obtained using both methodologies. In trial 1, the CH4 emissions were estimated from seven Holstein dairy cattle categories based on the SF6 tracer gas technique and on IPCC equations. The categories used in the study were prepubertal heifers (n=6); pubertal heifers (n=4); pregnant heifers (n=5); high-producing (n=6); medium-producing (n=5); low-producing (n=4) and dry cows (n=5). Enteric methane emission was higher for the category comprising prepubertal heifers when estimated by the equations proposed by the IPCC Tier 2. However, higher CH4 emissions were estimated by the SF6 technique in the categories including medium- and high-producing cows and dry cows. Pubertal heifers, pregnant heifers, and low-producing cows had equal CH4 emissions as estimated by both methods. In trial 2, two dairy farms were monitored for one year to identify all activities that contributed in any way to GHG emissions. The total emission from Farm 1 was 3.21t CO2e/animal/yr, of which 1.63t corresponded to enteric CH4. Farm 2 emitted 3.18t CO2e/animal/yr, with 1.70t of enteric CH4. IPCC estimations can underestimate CH4 emissions from some categories while overestimate others. However, considering the whole property, these discrepancies are offset and we would submit that the equations suggested by the IPCC properly estimate the total CH4 emission and carbon balance of the properties. Thus, the IPCC equations should be utilized with caution, and the herd composition should be analysed at the property level. When the carbon stock in pasture and other crops was considered, the carbon balance suggested that both farms are sustainable for GHG, by both methods. On the other hand, carbon balance without carbon stock, by both methods, suggests that farms emit more carbon than the system is capable of stock., (Copyright © 2016 Elsevier B.V. All rights reserved.)

An ethnopharmacological survey indicates that the genus Byrsonima has some medicinal species that are commonly found in the Brazilian Cerrado and has been used as an anti-inflammatory and for gastroduodenal disorders. The aim of this study was to evaluate the anti-inflammatory and antioxidant activity along with qualitative chemical characterization of the methanolic extract of the leaves of Byrsonima verbascifolia (BvME) obtained by exhaustive percolation. The data from the chemical analyses by liquid chromatography-mass spectrometry led to tentative identification of 42 compounds belonging to proanthocyanidins, galloyl quinic acid derivatives, flavonoids, and triterpene glycoside derivatives. BvME contain flavonoids and show an antioxidative activity. The methanolic extract administered intraperitoneally at doses of 50, 100, or 300 mg/kg showed a significant reduction in paw edema and modulated the neutrophil influx in a mouse model. Furthermore, the anti-edematogenic activity of the extract provided in smaller doses (12.5 and 25 mg/kg) was also demonstrated in a mouse paw edema model. The extract inhibited NO production by macrophages induced by lipopolysaccharide. We presume that the anti-inflammatory effects of BvME are due to a combination of compounds present in B. verbascifolia, including catechins (procyanidins), flavonoids, and triterpene glycosides and that these anti-inflammatory actions should be mediated, at least partly, through the inhibition of NO production. This study supports and validates the ethnopharmacological uses of B. verbascifolia as an anti-inflammatory.

Spiders are small arthropods that have colonized terrestrial environments. These impose three main problems: (i) terrestrial habitats have large fluctuations in temperature and humidity; (ii) the internal concentration of water is higher than the external environment in spiders, which exposes them continually to water loss; and (iii) their small body size determines a large surface/volume ratio, affecting energy exchange and influencing the life strategy. In this review we focus on body design, energetic, thermal selection, and water balance characteristics of some spider species present in Chile and correlate our results with ecological and behavioral information. Preferred temperatures and critical temperatures of Chilean spiders vary among species and individuals and may be adjusted by phenotypic plasticity. For example in the mygalomorph high-altitude spider Paraphysa parvula the preferred temperature is similar to that of the lowland spider Grammostola rosea; but while P. parvula shows phenotypic plasticity, G. rosea does not. The araneomorph spiders Loxosceles laeta and Scytodes globula have greater daily variations in preferred temperatures at twilight and during the night, which are set to the nocturnal activity rhythms of these species. They also present acclimation of the minimum critical temperatures. Dysdera crocata has a low preferred temperature adjusted to its favorite prey, the woodlouse. Spider metabolic rate is low compared to other arthropods, which may be associated with its sit and wait predatory strategy particularly in primitive hunter and weavers. In mygalomorph spiders the respiratory system is highly optimized with high oxygen conductance, for example G. rosea needs only a difference of 0.12-0.16 kPa in the oxygen partial pressure across the air-hemolymph barrier to satisfy its resting oxygen consumption demands. Water loss is a significant stress for spiders. Paraphysa parvula shows an evaporative water loss 10 times more than usual when the temperature approaches 40°C and the participation of book lungs in this loss is about 60%. This species and others show seasonal changes in water loss accounted for by changes in cuticle permeability. The case of Chilean spiders shows how the ecophysiology in spiders is associated to their design and body size and how is affected by fluctuating Mediterranean environments, suggesting that the adaptive process can be seen as a route of optimizing the use of energy to cope with environmental restrictions imposed by the interaction with the terrestrial environment and lifestyle.

Substances derived from plants play an important role in the development of new analgesic drugs, among them, triterpenoids. The connection between the participation of L-arginine/NO/cGMP pathway and the activation of ATP-sensitive K(+) channels (KATP) has been established on the peripheral antinociception induced by various drugs. The study assessed the involvement of L-arginine/NO/cGMP/KATP pathway in the antinociceptive effect induced by tingenone, from Maytenus imbricata, against the hyperalgesia evoked by prostaglandin E2 (PGE2) in peripheral pathway. The paw pressure test was used, with hyperalgesia induced by intraplantar injection of PGE2 (2 μg). Tingenone (200 µg/paw) administered into the right hind paw induced a local antinociceptive effect, that was antagonized by l-NOArg, nonselective nitric oxide synthase (NOS) inhibitor and by L-NPA, selective neuronal NOS (nNOS) inhibitor. The L-NIO, selective inhibitor of endothelial (eNOS), and the L-NIL, selective inhibitor of inducible (iNOS), did not alter the peripheral antinociceptive effect of the tingenone. The ODQ, selective soluble guanylyl cyclase inhibitor, prevented the antinociceptive effect of tingenone, and zaprinast, inhibitor of the phosphodiesterase that is cyclic guanosine monophosphate (cGMP) specific, intensified the peripheral antinociceptive effect of the smaller dose of tingenone. Glibenclamide, ATP-sensitive K(+) channels (KATP) blocker, but not tetraethylammonium chloride, voltage-dependent K(+) channel blocker; dequalinium dichloride, blocker of the small conductance Ca(2+)-activated K(+) channel, and paxilline, a potent blocker of high-conductance Ca(2+)-activated K(+) channels, respectively, prevented the peripheral antinociceptive effect of tingenone. The results demonstrate that tingenone induced a peripheral antinociceptive effect by L-arginine/NO/cGMP/KATP pathway activation, with potential for a new analgesic drug., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Insects exhibit three patterns of gas exchange: continuous (CoGE), cyclic (CGE) and discontinuous (DGE). In this work, we present the first record of a DGE in Phasmatodea and its transition to CGE and to CoGE through a thermal gradient. The rate of CO2 production (VCO2) at 10, 20 and 30°C was examined in adults of Agathemera crassa, a high-Andean phasmid of central Chile. Carbon dioxide release was recorded during 24 h with L:D cycle of 12:12 h in order to record both rest and activity periods. At rest, A. crassa showed three patterns of gas exchange, highlighting the use of DGE preferably at 10°C. As the temperature increased, the CoGE pattern was more frequent being the only pattern observed in all individuals at 30°C. During activity, patterns changed to CoGE with a significant increase in VCO2. Our results support the idea that gas exchange patterns in insects are not distinct but correspond to a continuum of responses addressed by metabolic demand and where DGE can be expressed only under an absolute state of rest. Our results support the idea that the presence of the DGE may be underestimated in other insect taxa because they may have been measured under conditions where this pattern not necessarily can be expressed., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Aim: To review the results of implementing a protocol for following up patients with idiopathic intracranial hypertension (IICH) in a neuro-ophthalmological unit (NOU)., Patients and Methods: A review of the literature was conducted in order to determine the examinations that needed to be included in the follow-up protocol, as well as the optimum frequency of visits and the most adequate duration of the follow-up. Later, a prospective review was performed of the patients that have been included since the NOU was set up and they were compared with the patients included in the IICH register prior to the creation of the NOU., Results: Since the protocol was implemented, visual acuity and the visual field have been evaluated in 100% of patients at three months, at six months and at one year after diagnosis. Moreover, the visual field was examined at three months, at six months and at one year after diagnosis in 91%, 72.8% and 100% of patients with IICH, respectively. Before our follow-up protocol was implemented, 190 had been carried out, which is roughly three per patient. Eleven lumbar punctures have been performed since the NOU was set up., Conclusions: The creation of a multidisciplinary NOU makes it possible to optimise resources and improve the care given to patients with IICH. This should result in an improvement in the functional prognosis of these patients.

Aim: The present study investigates the interaction of TLR4 and RAGE with their respective ligands as inducers of the inflammatory markers IL-6 and TNF-α. Also, the reactivity of peripheral blood mononuclear cells (PBMNC) from type 2 diabetic (T2D) patients and non-diabetic healthy controls (ND) were comparatively studied., Methods: Concentrations of IL-6 and TNF-α were measured by sandwich Elisa, using kits supplied by Assay Designs (Ann Arbor, MI, USA). PBMNC from T2D and ND were incubated in the presence or absence of LPS, anti-TLR4 or anti-RAGE for 72 hours at 37°C under 5% CO(2). The final volume was adjusted to 300 μL in DMEM supplemented with 10% fetal bovine serum. After incubation, the cells were centrifuged, the supernatant collected and the cytokines measured., Results: PBMNC from T2D were more sensitive to innate immune stimulation with LPS and monoclonal agonist anti-TLR4 than were cells from ND. The actions of LPS, anti-TLR4 and anti-RAGE potentiated the production of IL-6 and TNF-α in both groups. The simultaneous activation of monoclonal anti-RAGE and anti-TLR4 suggests that both antibodies used different receptors on the cell surface, but converged on the same PBMNC signaling metabolic pathways. This simultaneous activation induced a higher production of IL-6 and TNF-α in PBMNC from the T2D patients than from the ND subjects., Conclusion: Our results clearly show an exacerbation of innate immunity in PBMNC with T2D that was possibly hyperglycaemia-induced. These data, when analyzed together, suggest the importance of innate immunity in the pathogenesis of T2D., (Copyright © 2011 Elsevier Masson SAS. All rights reserved.)

Our planet is undergoing fast environmental changes, which are referred as global change. In this new scenario, it is of paramount relevance to understand the mechanistic basis of animal responses to environmental change. Here we analyze to what extent seasonal changes in the digestive function of the lizard Liolaemus moradoensis is under endogenous (i.e., hard wired) or exogenous (i.e., environmentally determined) control. For this purpose we compared animals collected in the field during autumn, winter and summer, against (experimental) specimens collected in the field at the beginning of autumn and reared in the laboratory under simulated summer conditions until winter. We found that different aspects of the digestive function are under different types of control: small intestine length appears to be under endogenous control (i.e., experimental animals were similar to winter animals), small intestine mass appears to be under exogenous control (i.e., experimental animals were similar to summer animals), and specific enzyme activities did not change throughout the year. Thus, we suspect that processes related with gut length, such as cell division, may be under endogenous control, while others related with gut mass, such as enterocyte size and content, may be determined by exogenous factors, such as the presence of food in the intestinal lumen. Faced with accelerated changing conditions, the ability of vertebrates to cope will be closely related with their plasticity in fitness-associated traits. More studies aimed at determining the levels and limits of physiological flexibility will be necessary to understand this phenomenon., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The understanding of animal functioning in fluctuating environments is a major goal of physiological and evolutionary ecology. In temperate terrestrial habitats, one of the most pervasive changes in environmental conditions is that associated with the seasonal change along the year. In this study, we describe the pattern of seasonal variation in the size of nine internal organs in the lizard Liolaemus moradoensis from the Andes Mountains of Central Chile. We observed that the size of digestive organs was greater during summer in comparison to other seasons. Dry masses of liver and fat bodies reached maximum values during summer and minimum during spring. We suspect that lowest spring values are related with build-up costs of energetically expensive organs (e.g., digestive, muscle mass) at the end of the hibernation period. Dry mass of the heart and lungs did not show a clear pattern of variation, suggesting that cardiac and pulmonary performance were maintained throughout the year. The dry mass of kidneys was greater during winter than during summer, a result observed in other hibernating lizards but for which there is no clear explanation. Finally, the dry mass of testes showed a maximum value during autumn and a progressive reduction toward summer, indicating that reproduction occurs during autumn. When represented in a bivariate space, acquisition (digestive), distribution (heart, lungs and kidneys), storage (liver and fat bodies), and expenditure (testes) organs generate four clusters. In general terms, observed seasonal pattern of change in organ size is in agreement with those reported for other lizard species that inhabit highly fluctuating environments., (© 2010 Wiley-Liss, Inc.)

One of the most controversial hypotheses that associate basal metabolic rate (BMR) with food habits and habitat productivity is the food habit hypothesis (FHH). Here we examined the relationship between BMR, diet, and climate among populations of the omnivorous passerine, Zonotrichia capensis (Emberizidae). We used nitrogen stable isotopes to estimate each individual's relative trophic level. To tease apart the effect of climatic variables and diet on BMR, we also used structural equation modeling. After the effect of body mass and climatic variables was taken into account, a significant effect of trophic level as estimated by delta(15)N on BMR was found. Our result seems to support the FHH at the intraspecific level, i.e., birds from the lower trophic levels - feeding on seeds and bud - had higher BMR than individuals from higher trophic levels.

Introduction: Idiopathic intracranial hypertension (IICH) is characterized by a rising in intracranial hypertension without evidence of intracranial expansive disease. Diagnostic criteria of headache related to IICH are described in the International Classification of Headache Disorders. In clinical practice, however, headache related to IICH may be heterogeneous., Patients and Methods: Clinical charts of patients with IICH admitted to the Hospital Universitario de Gran Canaria Dr. Negrin between 1990 and 2007 were retrospectively reviewed., Results: Fifty-five patients were included, with a ratio female/male of 8.1:1. Headache was present in 85.4%. The headache was continuous in 63.8% of patients, and diffuse in 51%., Conclusion: Headache related to IICH is heterogeneous, and may mimic primary headache, so a high level of suspicion is needed to avoid diagnostic delay.

Hibernation in ectothermic animals was historically considered as a simple cold-induced torpor state resulting from the inability to maintain a high body temperature at low ambient temperatures. During the last decades this vision changed and nowadays there is a myriad of studies showing that hibernation implies different adjustments at the genetic, molecular, biochemical and cellular levels. However, studies oriented to evaluate changes of whole organism structure and physiology still are scarce, which is particularly true for amphibians that hibernate on land. Accordingly, in the Andean toad (Bufo spinulosus), we investigated the effect of short-term fasting and hibernation on the hydrolytic activity of digestive enzymes, histology of the small intestine, gross morphology of digestive and other internal organs and standard metabolic rate. Based on the pattern of size variation, internal organs may be grouped into those that were affected by both season and feeding condition (small intestine, stomach and liver), those that were only affected by season (fat bodies), those that were only affected by feeding condition (kidneys) and, finally, those that did not change between the three groups (large intestine, heart and lungs). Hydrolytic activity of maltase, trehalase and aminopeptidase-N followed the same pattern of variation (feeding > fasting > hibernating toads), although the change for the latter enzyme was less noticeable than for the disaccharidases. Enzymatic adjustments were correlated with changes in small intestine histology: villus and enterocyte height increased from hibernating to fasting and more markedly from fasting to feeding toads. Metabolic rate decreased during hibernation to 7.8% (at 5 degrees C) and 13.6% (at 15 degrees C) of summer values, which is one of the highest metabolic depressions reported for any ectothermic vertebrate. Our results suggest that amphibian persistence in highly seasonal environments is related to a large capacity of phenotypic flexibility at different organisational levels; an ability that may be related to the extensive ranges of temporal existence and geographic distribution of these vertebrates.

A large number of physiological acclimation studies assume that flexibility in a certain trait is both adaptive and functionally important for organisms in their natural environment; however, it is not clear how an organism's capacity for temperature acclimation translates to the seasonal acclimatization that these organisms must accomplish. To elucidate this relationship, we measured BMR and TEWL rates in both field-acclimatized and laboratory-acclimated adult rufous-collared sparrows (Zonotrichia capensis). Measurements in field-acclimatized birds were taken during the winter and summer seasons; in the laboratory-acclimated birds, we took our measurements following 4 weeks at either 15 or 30 degrees C. Although BMR and TEWL rates did not differ between winter and summer in the field-acclimatized birds, laboratory-acclimated birds exposed to 15 degrees C exhibited both a higher BMR and TEWL rate when compared to the birds acclimated to 30 degrees C and the field-acclimatized birds. Because organ masses seem to be similar between field and cold-acclimated birds whereas BMR is higher in cold-acclimated birds, the variability in BMR cannot be explained completely by adjustments in organ masses. Our findings suggest that, although rufous-collared sparrows can exhibit thermal acclimation of physiological traits, sparrows do not use this capacity to cope with minor to moderate fluctuations in environmental conditions. Our data support the hypothesis that physiological flexibility in energetic traits is a common feature of avian metabolism.

One of the most fundamental questions in organismal ecology is how animals work in a continuously changing environment. In order to contribute to the current understanding of this question, this study evaluated seasonal changes in digestive enzymes activities, organs size, and energy reserves in Liolaemus nigroviridis, a medium-size lizard that inhabit extreme environments in the Andes range. We found that digestive enzymes (trehalase, maltase, and aminopeptidase-N) hydrolytic activities, dry masses of digestive organs and liver, and energy reserve (dry mass of fat bodies and tail energy density) were greater during summer than during winter months. By contrast, dry mass of the kidneys, lungs, heart, and gonads were greater during winter (though significance was reach only for the last two organs). In summary, obtained results reinforce the idea that hibernation is connected with phenotypic adjustments at different organizational levels, which in turn, potentially affects rates of energy acquisition and expenditure, organisms' fitness, and, ultimately, ecological and evolutionary success of species living in highly seasonal environments. We suggest that, owing to the pressing need to explain and predict the impact of climatic change on the biota, more studies destined to determine the levels and limits of physiological flexibility are necessary., ((c) 2009 Wiley-Liss, Inc.)

According to the "barrel model", an organism may be represented by a container, with input energy constraints (foraging, digestion, and absorption) symbolized by funnels connected in tandem, and energy outputs (maintenance, growth, and reproduction) symbolized by a series of spouts arranged in parallel. Animals can respond to changes in environmental conditions, through adjustments in the size of the funnels, the fluid stored inside the barrel, or the output flow through the spouts. In the present study, we investigate the interplay among these processes through the analysis of seasonal changes in organ size and metabolic rate in a lizard species (Liolaemus bellii) that inhabits extremely seasonal environments in the Andes range. We found that digestive organ size showed the greatest values during spring and summer, that is, during the foraging seasons. Energy reserves were larger during summer and autumn, and then decreased through winter and spring, which was correlated with overwintering maintenance and reproductive costs. Standard metabolic rate was greater during the high-activity seasons (spring and summer), but this increase was only noticeable at higher environmental temperatures. The ability of many lizard species to reduce their maintenance cost during the cold months of the year, beyond what is expected from temperature decrease, is probably related to their success in coping with highly fluctuating environments. Here, we demonstrate that this ability is correlated with high physiological flexibility, which allows animals to adjust energy acquisition, storing and expenditure processes according to current environmental conditions.

Objective: To compare the role of Akt/PKB signaling pathway in the modulation of reactive oxygen species (ROS) production by autologous plasma in peripheral blood mononuclear cells (PBMNC) from type 2 diabetic patients and healthy subjects., Materials and Methods: This study was approved by Santa Casa Ethical Committee and has included patients diagnosed with diabetes type 2 (DM2) and control group (non-diabetic) (ND). PBMNC were purified utilizing Ficoll-hypaque gradient. ROS was quantified by luminol-dependent chemiluminescence. The Akt/PKB phosphorylation was measured using a CASE kit. Statistical analyses were made with t Student test and chi-square (chi(2)). p<0.05 was considered significant., Results: 12, 13-Phorbol dibutyrate (PDB) stimulated the production of higher levels of ROS in PBMNC from type 2 diabetic patients than that from healthy subjects. Autologous plasma, however, inhibited induced or not ROS production in PBMNC in both groups. The inhibition of PBMNC-ROS derived by autologous plasma from healthy subjects was higher than that from type 2 diabetic patients. Plasma phosphorylated (activated) Akt/PKB. The percentage of phosphorylation induced by autologous plasma in PBMNC from patients and healthy control were 14% and 93%, respectively. Inhibition of ROS production in PBMNC from DM2 were similar for PBMNC+plasma; PBMNC+Akti; and PBMNC+plasma+Akti. However, in ND control, plasma showed a higher ROS inhibition than Akti or plasma plus Akti., Conclusions: Our results suggest that the low antioxidant capacity observed in autologous plasma from DM2 patients in conjunction with the decreased activation of PKB may cause an imbalance in the oxidizing/reducing responses, possible inducing an oxidative stress state, which could be associated with tissular damage.

The bronchial tree of most mammalian lungs is a good example of an efficient distribution system whose geometry and dimensions of branched structures are important factors in determining the efficiency of respiration. Small and flying endothermic animals have high-energy requirements, requiring morphological and physiological adaptations to reduce energy loss. Here we show that Tadarida brasiliensis, a nocturnal small bat whose energy requirements are exacerbated by this small size and by their frequent exposure to high altitude, has a different morphology in the proximal airway, sustained by a wider trachea and better scaling factors, than other non-flying mammals. This design allows a great decrease of the volume specific resistance of the proximal airway and in consequence a very low entropy production during breathing, approximately 1/18 of that expected for a non-flying mammals of similar body size.

Introduction: Cerebral venous sinus thrombosis (CVSTs) constitutes an infrequent location of the venous thrombotic disease. It is caused by diverse factors, although up to 35% of the CVSTs can remain without aetiologic diagnosis. Coagulation disorders have been involved in up to 75% of the CVSTs. AIM. To describe the prevalence of coagulation disorders in a consecutive series of CVSTs., Patients and Methods: We performed a retrospective revision of the clinical history of all patients with CVST admitted in our service between January, 1st 1995 and December, 31st 2005. The thrombophilic study was carried out a posteriori in whatever individual who lacked it., Results: We studied twenty-one cases (7 men and 14 women), within an age range of 16-69 years old (corresponding to an average of 35,4 +/- 14,4 years old). The thrombophilic study was carried out in eighteen patients. Five (27,7%) presented some hereditary coagulation disorder; in four of them, a hormonal cause coexisted., Conclusion: Recent advances in the diagnostic procedures for the detection of thrombophilias and hereditary prothrombotic mutations have enabled the specialists to be more precise in the aetiologic diagnosis of suspected CVST. Even so, the apparent cause for a high percentage of patients with CVST has not been found yet.

The construction of a realistic theoretical model of proteins is determinant for improving the computational simulations of their structural and functional aspects. Modeling proteins as a network of non-covalent connections between the atoms of amino acid residues has shown valuable insights into these macromolecules. The energy-related properties of protein structures are known to be very important in molecular dynamics. However, these same properties have been neglected when the protein structures are modeled as networks of atoms and amino acid residues. A new approach for the construction of protein models based on a network of atoms is presented. This method, based on interatomic interaction, takes into account the energy and geometric aspects of the protein structures that were not employed before, such as atomic occlusion inside the protein, the use of solvation, protein modeling and analysis, and the use of energy potentials to estimate the energies of interatomic non-covalent contacts. As a result, we achieved a more realistic network model of proteins. This model has the virtue of being more robust in face of different unknown variables that usually are arbitrarily estimated. We were able to determine the most connected residues of all the proteins studied, so that we are now in a better condition to study their structural role.

We propose a novel method for defining patterns of contacts present in protein-protein complexes. A new use of the traditional contact maps (more frequently used for representation of the intra-chain contacts) is presented for analysis of inter-chain contacts. Using an algorithm based on image processing techniques, we can compare protein-protein interaction maps and also obtain a dissimilarity score between them. The same algorithm used to compare the maps can align the contacts of all the complexes and be helpful in the determination of a pattern of conserved interactions at the interfaces. We present an example for the application of this method by analyzing the pattern of interaction of bovine pancreatic trypsin inhibitors and trypsins, chymotrypsins, a thrombin, a matriptase, and a kallikrein - all classified as serine proteases. We found 20 contacts conserved in trypsins and chymotrypsins and 3 specific ones are present in all the serine protease complexes studied. The method was able to identify important contacts for the protein family studied and the results are in agreement with the literature.

Lizard tail autotomy is considered an efficient anti-predator strategy that allows animals to escape from a predator attack. However, since the tail also is involved in many alternative functions, tailless animals must cope with several costs following autotomy. Here we explicitly evaluate the consequences of tail autotomy for two costs that have been virtually unexplored: 1. we test whether the anatomical change that occurs after tail loss causes a reduction in the role of the tail as a distraction mechanism to predators; 2. we analyzed whether tail synthesis comprises an energetically costly process in itself, by directly comparing the cost of maintenance before and after autotomy. We found that original tails displace further and at greater velocity than regenerated tails, indicating that the anti-predation responses of a lizard probably changes according to whether its tail is original or regenerated. With regard to the energetic cost of tail synthesis, we observed a significant increase in the standard metabolic rate, which rose 36% in relation to the value recorded prior to tail loss. This result suggests that the energetic cost of tail synthesis itself could be enough to affect lizard fitness.

Oxidizing/reducing response by granulocytes and a potential correlation between reactive oxygen species generation and triglyceride, total cholesterol, low-density lipoprotein, high-density lipoprotein, body mass index, fasting plasma glucose, glycemic control (hemoglobin A(1c)), or duration of diabetes were examined in type 2 diabetic patients and in healthy subjects. An increase in both oxidizing and reducing responses was observed in cells from diabetic patients relative to normoglycemic individuals. The increase in oxidizing response was nearly 2-fold higher, whereas the antioxidant response increased by 50%. Although reactive oxygen species generation from healthy subjects was correlated with levels of low-density lipoprotein (positive correlation), high-density lipoprotein (negative correlation), and body mass index (positive correlation), no such associations were observed in diabetic subjects, suggesting either an intrinsic perturbation in the oxidant/antioxidant response or possibly due to the effect of the various medications being taken by these patients. These issues need further study.

We tested the hypothesis that women with greater prenatal maternal stress (PNMS) would be more likely to receive intravenous opiates and epidural for delivery, and thereby increase the likelihood of unplanned cesarean delivery. PNMS was assessed during early, mid, and late pregnancy using psychometrically sound instruments in structured interviews with women receiving prenatal care at a public university clinic. Medical records were abstracted for analgesia during delivery, fetal heart tracing (FHT) abnormalities, and method of delivery. Only subjects attempting vaginal delivery (N = 298) were included. Using structural equation modeling, a PNMS variable was constructed from five indicators: pregnancy-specific distress, number of prenatal stressful life events, distress from life events, state anxiety, and perceived stress. After controlling for medical predictors of analgesia receipt and surgical delivery, women with higher PNMS were more likely to receive analgesia, and those who received analgesia were more likely to deliver surgically. Analgesia was also associated with FHT abnormalities, which in turn was associated with surgical delivery (all p's < 0.05). Women who received both an epidural and meperidine were most likely to have a cesarean delivery; 29% of this group delivered surgically. Results indicate that PNMS contributes to higher likelihood of unplanned cesarean delivery through its association with delivery analgesia.

We investigated the intraspecific variation in basal metabolic rate (BMR) and total evaporative water loss (TEWL) in the omnivorous passerine Zonotrichia capensis from two populations inhabiting regions with different precipitation regimes and aridity indices. Values of TEWL in birds from the semi-arid region were significantly lower than those found in sparrows from the mesic region. TEWL in birds from the semi-arid site was 74% of the expectation based on body mass for passerines from mesic areas and similar to the allometric expectation for passerines from arid environments. In sparrows from the mesic area, TEWL was higher than predicted by their body mass for passerines from arid environments (133%), but very close (97%) to the expectation for passerines from mesic areas. BMR values were 25% lower in sparrows from the semi-arid region. The lower TEWL and BMR of birds from the semi-arid region may be a physiological adjustment that allows them to cope with fewer resources and/or water. We propose that the lower endogenous heat production in birds from the semi-arid environment may decrease their water requirements.

Thirty, 4-month-old entire Santa Ines lambs were grazed on an Andropogon gayanus pasture, during a 34-week period (rainy season weeks 0-20 and dry season weeks 21-34) and allocated in two treatment groups (n = 15) each with different protein supplementation: high protein (HP-19% CP) and low protein (LP-11% CP). These were subdivided into those receiving anthelmintic treatment (c) (n = 7) and without anthelmintic treatment (i) (n = 8). The objective was to evaluate the effects of supplementation with protein on resistance and resilience to natural helminth infection of hair breed lambs. Lamb weight, blood collection and faecal egg counts (FEC) were carried out monthly. The lambs were slaughtered after 34 weeks, when worm burdens, worm length and eosinophil cell counts were taken. The sheep on treatments HPc and HPi were heavier in live weight than those from LPi and LPc (P < 0.05) at the end of the rainy period. The HPc group finished heavier (P < 0.05) than the other groups in the dry season, which had no significant differences between them. The predominant species of nematode found was T. colubriformis followed by H. contortus, Trichuris globulosa and Moniezia expansa. Animals on HPi had lower FEC than LPi (P < 0.05). The number of worms was lower for both HP groups (P < 0.05) with worm length shorter in the HPc group (P < 0.05) compared with all other groups. The number of eosinophils was higher in animals in the LPi group, which also showed anaemia and lower plasma urea at the end of the dry season. Diet supplementation with high protein was able to improve resilience and resistance to natural infection by endoparasites during the rainy season. In the dry season there was a decrease in both of these traits, which were intimately linked to the quality of available forage under tropical conditions.

Introduction: Pulmonary hypertension (PH) (mean pulmonary arterial pressure [mPAP] > 25 mm Hg) is frequently observed during the postoperative period after liver transplantation (LT)., Objective: The objective was to compare respiratory function, intensive care unit (ICU) length of stay (LOS), and 30-day survival rates among patients evolving with PH with those who do versus do not develop it during the postoperative period after LT., Methods: Fifty-seven patients undergoing LT from January 1999 to December 2000 were divided into 2 groups: Group 1 (G1; n = 26), without PH; and Group 2 (G2; n = 31), with moderate PH. Preoperative parameters were Child-Pugh's classification, pulmonary function tests, mPAP, and P(A-a)O(2). During the intraoperative period, warm and cold ischemic times and the amount of blood transfusion were evaluated, whereas mPAP, PaO(2)/FiO(2) ratio, weaning time, ICU LOS, and 30-day survival rates were evaluated postoperatively., Results: mPAP in early postoperative period was 21 +/- 13 mm Hg and 32 +/- 4 mm Hg in G1 and G2, respectively (P <.0001). PaO(2)/FiO(2) was 310 +/- 82 mm Hg in G1 and 272 +/- 84 mm Hg in G2 (P =.48). In G1 and G2, 77% and 74% of patients, respectively, were successfully weaned in the first 24 hours postoperative (P =.10). ICU LOS was 111 hours (range, 45-1098 hours) in G1 and 102 hours (range, 59-284 hours) in G2 (P =.36). The 30-day survival rate was 20 of 26 (77%) in G1 and 26 of 31 (84%) in G2 (P =.44)., Conclusion: Our data suggest that moderate PH during the early postoperative phases of LT cannot be considered an additional risk factor for pulmonary dysfunction, and for an increased ICU LOS or 30-day mortality rate.

Animals process and allocate energy at different seasons at variable rates, depending on their breeding season and changes in environmental conditions and resulting physiological demands. Overall total energy expenditure, in turn, should either increase in some seasons due to special added demands (e.g. reproduction) or it could simply remain at about the same level, in which case the animals must show compensatory rebalancing of other expenditures that can be reduced. To test for the alternative hypotheses of seasonal variability or compensation, we measured total daily energy expenditure (DEE) in free-living degus (Octodon degus) at four seasons and followed this with determinations of basal metabolic rate (BMR) in the laboratory in the same individuals. DEE varied seasonally but was only significantly different (lower) in summer (non-breeding season), with a DEE:BMR ratio of only 1.6, whereas autumn, winter and spring DEE values were statistically indistinguishable from one another and showed DEE:BMR ratios ranging from 1.9 to 2.2. Our values of DEE in the field fall within the broad range of allometric expectation for herbivorous mammals in general, but the ratios of DEE:BMR are lower than expected. This, together with the lack of strong major shifts in total levels of DEE, suggests that degus are showing compensatory shifts among various categories of energy expenditure that allow them to manage their overall energy balance by minimizing total expenditure.

Milk is the sole food source of mammals during early postnatal development, and its composition may be modified to meet changing nutritional and energetic demands from birth to weaning. These demands are especially acute in small mammals that breed in highly seasonal environments. We investigated the temporal course of milk composition during lactation in free-living yellow-pine chipmunks, Tamias amoenus, a small altricial omnivorous rodent that produces a single annual litter immediately after emerging from hibernation. Over the course of lactation the total energy concentration of milk increased more than two-fold, to approximately 13 kJ ml(-1). The main component of the milk was lipids, which increased from 10 to 30% of total milk content by wet mass. Proteins increased from approximately 5 to 10%, whereas carbohydrates remained low, at 4-5%. The progressive augmentation of milk energy content during lactation contributes to the increased demands of rapid growth in this altricial species.

We studied the ontogeny of the intestinal brush border disaccharidases sucrase and lactase in the precocial rodent Octodon degus. Sucrase hydrolyze sugars from plants while lactase hydrolyzes sugars from milk. Enzyme expression varied inversely with dietary changes according to the developmental pattern. All new-born pups had high lactase and low sucrase activities. Also, a negative correlation between sucrase and lactase activity was found, supporting the economic design hypothesis for the intestinal tract. Profiles for development of sucrase expression exhibit some differences among precocial species, and in O. degus is correlated with the slower transition from milk to solid food consumption at weaning.

The onset of mating in yellow-pine chipmunks (Tamias amoenus) follows emergence from a prolonged period of energy conservation during hibernation. Energy expenditures are greatly accelerated to meet the demands of the reproductive season. When emerging from hibernation, typical male chipmunks (breeders) have enlarged testes and a high level of plasma testosterone (T). However, certain males that do not participate in reproduction (nonbreeders) maintain small testes and low plasma T levels and emerge several weeks later than the breeders. The timing of the terminal arousal from hibernation and onset of mating are associated with increased plasma T levels. Experimental elevation of T levels in T. amoenus outside the mating season has been associated with a decrease in body mass, further suggesting an effect of T on energy balance. To test this hypothesis, we measured daily energy expenditure (DEE) in free-living, nonbreeding male chipmunks in the presence and absence of a T-implant. We also measured DEE in breeding males when endogenous T levels were high. DEE of the nonbreeders was not affected by our manipulation of plasma T, and the DEE of breeding males did not differ from that of nonbreeders. We conclude that energy expenditure on a daily basis in male yellow-pine chipmunks is not influenced by levels of T. However, on a seasonal basis, the earlier emergence from hibernation by breeding males, which appears to be influenced by T, represents an overall seasonal energy expenditure that exceeds that of nonbreeding males., (Copyright 2002 Wiley-Liss, Inc.)

We investigated seasonal patterns of plasma glucocorticosteroids (GCs) in both sexes and testosterone (T) in males in relation to the annual cycle in central Chile of a natural population of the degu (Octodon degus), a caviomorph rodent. We wanted to find out which GCs are present in degus, whether their seasonal variation suggests suppressive or synergistic interrelationships with T, and whether seasonal variation in GC levels indicates a relationship with energy mobilization and demands of reproduction. Degus mated in late autumn, and female body mass increased in pregnancy and remained high during lactation and throughout spring. Over the subsequent period of summer drought both sexes declined to a minimal body mass before the next mating season. Cortisol appears to be the principal GC in degus. In fact cortisol levels were so high that the extremely low levels of corticosterone measured were probably largely due to the cross-reactivity of our corticosterone antiserum with cortisol. Titers of cortisol in females exceeded 1000 ng/ml at lactation in the spring of 2 years; cortisol declined greatly following lactation and during the summer and reached its lowest mean level of about 500 ng/ml at mating. Males were more difficult to capture than females and thus our sampling was limited, but male cortisol levels were similar to those of females during the times of year when we measured them. Male T levels remained within a low range all year, but at mating, when mean T was highest (0.16 ng/ml) and when most males had detectable T, degus showed their lowest cortisol levels. The minimal cortisol level of males during mating represents a possible suppressive effect of T, as described in other mammals. At the time of their spring emergence, 60% of juvenile males had detectable T levels comparable to those of adults, suggesting important organizational effects of T at that time in their maturation. Peak cortisol titers in both sexes were associated with lactation in females, when energy mobilization, production, and body mass were at their greatest., (Copyright 1999 Academic Press.)

Objective: To find the differences between telephone and mail interview respect on answer rate, economic cost and characteristics of participants. To determine the knowledge of our diabetics about their disease., Design: A descriptive transversal study., Setting: Primary care Centro de San Gregorio, Telde, Las Palmas., Patients: The diabetics type 1 and 2 (n = 566) of our centre, whose ages were 18-70 years. In order to apply the same health questionnaire through two different ways, we divided the sample into two random groups: mail group (n = 283) and telephone group (n = 283)., Results: The global answer rate was 51.6% (292 patients), 48% in the mail group (136 patients) and 55% in the telephone group (156 patients). The total cost of the mail survey was 41,000 pts. (104 pts. per mail), and the cost of telephone survey was 5.100 pts. (25.5 pts. per useful call). In the whole sample, we observed women more frequently in the answer group (56% of 292) than in the no answer group (44% of 274) (CI 95% 8.1-12.8). Also, in the answer group, the age was greater than in the no answer group (58.8 +/- 9.8 yr vs. 55.8 +/- 12.5 yr) (CI 95% 1.14-4.8). When we analyzed gender and age in each group, we observed the same differences in the telephone group but not in the mail group. More than the half questionnaire were correctly answer by 64% (167 patients). In the telephone group the "fit" rate was greater (70.6% of 153) than in the mail group (55.7% of 106) (CI 95% 3.2-26.6)., Conclusions: A suitable answer rate was reached through both methods of interview, but telephone survey's was smaller. In the mail group, both genders and all ages groups answered in the same way. The knowledge of our diabetics about their disease was like as the found in another studies.

Animals must match their foraging and digestion to seasonal changes in availability and quality of food. When these parameters decline, the animal's performance limits for extracting energy and nutrients may be challenged. In the laboratory, we investigated daily patterns of food processing on a low-quality (high-fiber) diet of alfalfa in an herbivorous, day-active rodent, the degu (Octodon degus), which inhabits semiarid central Chile. We manipulated timing of food availability, from continuous availability down to as little as 5 h/d. Degus maintained weight while digesting only 53% of dry-matter consumption. With food continuously available in a metabolic cage, the animals ate more food and deposited about twice as much feces in the day as at night. Continuous 24-h behavioral observation revealed that degus were actually defecating at the same rate both night and day but then ingesting most of the feces they produced at night. Further experimental treatments challenged animals with limited periods of food availability that matched natural foraging patterns. With either 11 h of daytime food availability or only 5 h (in morning and afternoon periods of 2.5 h each), degus consumed as much food as those with 24-h food availability. Continuous 24-h behavioral observations revealed in the 11-h group that nearly all feces produced at night were reingested and nearly none were reingested in the day, whereas the 5-h group resorted to further coprophagy during the 6-h midday interval with no food. Despite these differences in timing of food intake and coprophagy in response to the three experimental treatments, the degus were defecating at the same rate both night and day, which indicated a constant rate of output from the colon. This suggests a range of adjustments of digestive physiology to the timing of gut function by balancing coprophagy with ingestion of food. Overall, 38% of 24-h feces production was reingested, and 87% of this coprophagy occurred at night. The ingestion of feces during parts of the day when food is unavailable provides for continued intake into the digestive tract and appears to represent an increase in overall efficiency of gut use.

A full-length DNA clone encoding a putative pyruvate dehydrogenase alpha subunit (E1 alpha) gene was isolated from a Trypanosoma cruzi (RA strain) DNA library. Sequencing of this clone revealed it to encode a 378 amino acid protein (M(r) 42774) with high sequence similarity to E1 alpha obtained from different sources. The highest score is obtained with human E1 alpha: 43,3% similarity. Southern blot analysis is consistent with the existence of a single copy of this putative T. cruzi E1 alpha gene per haploid genome in different parasite strains. Expression of this gene was demonstrated by Northern blot analysis and its trans-splicing acceptor site was identified by Polymerase Chain Reaction-mediated amplification of its cDNA.

The present study compares the early diagnosis of congenital Chagas' disease with a DOT assay using recombinant antigens with immunofluorescence antibody testing (IFAT) and an enzyme-linked immunosorbent assay (ELISA). The studies were performed using cord blood and sera of 12 infected newborns (group I) and 12 uninfected ones born to Trypanosoma cruzi-infected mothers (group II). Conventional IFAT and ELISA showed positive results for IgG at high titers, in infants and mothers of both groups; IgA antibodies were detected by ELISA in four of the infected infants and IgM was detected in two of them. All sera of the uninfected infants were negative for IgA and IgM in the ELISA. Application of a DOT assay using eight recombinant T. cruzi antigens allowed detection of specific IgA in the cord blood of six of the infected cases and IgM in eight of them. Repetition of these serologic tests in samples obtained during a monthly follow-up gave positive results for IgA in two of the initially negative infants of group I and for IgM in four of them. This means that diagnosis of congenital T. cruzi infection was confirmed, through demonstration of specific IgM, in all infected infants, and of IgA in eight of them. The importance of late detection of IgM in siblings born of infected mothers is discussed. The detection of IgM and IgA in sera obtained after birth is believed to be due to a congenital transmission of the parasite that occurred late in pregnancy. No IgA or IgM antibodies could be detected by the DOT assay in the sera of the negative controls.(ABSTRACT TRUNCATED AT 250 WORDS)

A panel of eight recombinant Trypanosoma cruzi antigens was used to study the IgG reactivity of sera proceeding from 151 patients with chronic Chagas disease. These samples were reactive against T Cruzi epimastigotes by immunofluorescence and enzyme linked assays. A dot blot assay was performed to evaluate the reactivity against recombinant clones 1, 2, 13, 26, 30, 36, 54 and SAPA (Shed Acute Phase Antigen). Ninety six percent of samples reacted against one or more of these antigens. The most frequently recognized proteins were clones 1, 2, 13, 36 and 26 with 90, 89, 76, 71 and 66 percent of reactivity respectively. Less than 50% of sera reacted against clones 30, 54 and SAPA. Sera from symptomatic patients had a significantly higher frequency of reactivity against clone SAPA than those from asymptomatic patients. This difference suggests that a high reactivity against clone SAPA may be related to acute infections and symptoms. No significant relationship could be established between specific recombinant clone recognitions and particular clinical forms of the disease (cardiological or digestive). It is concluded that these recombinant proteins may become a good tool for the evaluation of the immune response of patients infected with T. cruzi.

A panel of eight Trypanosoma cruzi antigens produced by recombinant DNA techniques was used to compare the reactivity of IgG specificities in the sera from 45 chronic Chagas' disease patients with different clinical symptoms (cardiac disease, gastrointestinal lesions, and combined syndrome) with those present in the sera from 55 asymptomatic patients in Chile. All of the serum samples were first characterized for antibody to T. cruzi epimastigotes by immunofluorescence assay. All of the Chagas' disease sera were reactive, but none of five healthy controls whose sera were also tested had antibodies against the fixed parasites. A dot-blot assay was then performed to evaluate the serum reactivity against recombinant DNA clones 1, 2, 13, 26, 30, 36, 54, and SAPA (shed acute phase antigen). These recombinant antigens were recognized by a large proportion of the sera collected from the Chilean patients. Ninety-five percent of the serum samples reacted with one or more of the recombinant clones. Analysis of the reactivity with individual fusion proteins showed that 88% of these sera reacted with clones 1 and 2, and 78% reacted with clone 13. Differences in reactivity to clones number 13, 30, and SAPA were observed when symptomatic and asymptomatic patients were compared. These differences in reactivity were statistically significant (P less than 0.01) according to Fisher's exact test.

An enzyme-linked immunosorbent assay (ELISA) has been developed to detect antibodies in human sera to synthetic peptides derived from the repeating amino acid sequence in recombinant Trypanosoma cruzi antigens. Sixty serum samples from patients with chronic Chagas' disease were used to determine the reactivity against the synthetic repeat peptides derived from clones 1, 2, 30, 36, and shed acute phase antigen (SAPA). Ninety-eight percent of the samples had detectable antibodies to one or more of the synthetic peptides at titers greater than 1:100. The percentage of reactive sera increased from 28% with peptide SAPA to 93% with peptide 2. The exposure of patients to T. cruzi was reflected in indirect immunofluorescent antibody titers to fixed epimastigotes. Comparisons between ELISA and immunoradiometric assay data indicated that both tests were of approximately equal sensitivity.