Standardisation of Western blotting to detect HTLV-1 antibodies synthesised in the central nervous system of HAM/TSP patients

Intrathecal synthesis of human T-lymphotropic virus type 1 (HTLV-1) antibodies (Abs) represents conclusive evidence of a specific immune response in the central nervous system of HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients. Western blotting (WB) for HTLV Abs in serum is a confirmatory test for HTLV-1 infection. The aim of this study was to standardise the Western blot to demonstrate the intrathecal pattern of Abs against HTLV-1 proteins in HAM/TSP patients. Paired cerebrospinal fluid (CSF) and serum samples were selected from 20 patients with definite HAM/TSP, 19 HTLV-1 seronegative patients and two HTLV-1 patients without definite HAM/TSP. The presence of reactive bands of greater intensity in the CSF compared to serum (or bands in only the CSF) indicated the intrathecal synthesis of anti-HTLV-1 Abs. All definite HAM/TSP patients presented with an intrathecal synthesis of anti-HTLV-1 Abs; these Abs were not detected in the control patients. The most frequent intrathecal targets of anti-HTLV-1 Abs were GD21, rgp46-I and p24 and, to a lesser extent, p19, p26, p28, p32, p36, p53 gp21 and gp46. The intrathecal immune response against env (GD21 and rgp46-I) and gag (p24) proteins represents the most important humoral pattern in HAM/TSP. This response may be used as a diagnostic marker, considering the frequent association of intrathecal anti-HTLV-1 Ab synthesis with HAM/TSP and the pathogenesis of this neurological disease.     

Serologic survey of West Nile virus in horses from Central-West,Northeast and Southeast Brazil

Since the emergence of West Nile virus (WNV) in North America in 1999, there have been several reports of WNV activity in Central and South American countries. To detect WNV in Brazil, we performed a serological survey of horses from different regions of Brazil using recombinant peptides from domain III of WNV. Positive samples were validated with the neutralisation test. Our results showed that of 79 ELISA-positive horses, nine expressed WNV-specific neutralising antibodies. Eight of the infected horses were from the state of Mato Grosso do Sul and one was from the state of Paraíba. Our results provide additional evidence for the emergence of WNV in Brazil and for its circulation in multiple regions of the country.     

Seasonal Habitat Use of Agoutis (Dasyprocta azarae) is Driven by the Palm Attalea phalerata in Brazilian Pantanal

Resource availability can influence animal movement causing changes in home‐range size and arrangement between seasons. We investigated the influence of acuri palm (Attalea phalerata) on the occupancy of agouti Dasyprocta azare during both the dry and rainy seasons, as well as the abandonment probabilities in the transition between seasons in the Brazilian Pantanal. The agoutis occupied a high proportion of the forested areas in the rainy season (0.83), but the occupancy decreased in the dry season (0.39). In the rainy season, occupancy by agoutis was not correlated with acuri palm availability, while in the dry season, it was positively correlated with the palm's availability. The acuri palm availability also drove the agoutis' probability of site abandonment from the rainy to the dry season, with higher abandonment probability in sites with low acuri palm availability. These findings show that this large‐seeded palm can be an important resource for the agoutis' populations during the fruit‐scarcity period. The acuri palm seeds may be particularly important for agoutis using fragmented forests, given that the large‐seeded palms are able to persist even in small fragments, which is untrue for other important resource.     

XX/XO, a rare sex chromosome system in Potamotrygon freshwater stingray from the Amazon Basin, Brazil

Potamotrygonidae is a representative family of South American freshwater elasmobranchs. Cytogenetic studies were performed in a Potamotrygon species from the middle Negro River, Amazonas, Brazil, here named as Potamotrygon sp. C. Mitotic and meiotic chromosomes were analyzed using conventional staining techniques, C-banding, and detection of the nucleolus organizing regions (NOR) with Silver nitrate (Ag-NOR). The diploid number was distinct between sexes, with males having 2n = 67 chromosomes, karyotype formula 19m + 8sm + 10st + 30a, and fundamental number (FN) = 104, and females having 2n = 68 chromosomes, karyotype formula 20m + 8sm + 10st + 30a, and FN = 106. A large chromosome, corresponding to pair number two in the female karyotype, was missing in the male complement. Male meiotic cells had 33 bivalents plus a large univalent chromosome in metaphase I, and n = 33 and n = 34 chromosomes in metaphase II. These characteristics are consistent with a sex chromosome system of the XX/XO type. Several Ag-NOR sites were identified in both male and female karyotypes. Positive C-banding was located only in the centromeric regions of the chromosomes. This sex chromosome system, which rarely occurs in fish, is now being described for the first time among the freshwater rays of the Amazon basin.     

Fish oil supplementation for two generations increases insulin sensitivity in rats

We investigated the effect of fish oil supplementation for two consecutive generations on insulin sensitivity in rats. After the nursing period (21 days), female rats from the same prole were divided into two groups: (a) control group and (b) fish oil group. Female rats were supplemented with water (control) or fish oil at 1 g/kg body weight as a single bolus for 3 months. After this period, female rats were mated with male Wistar rats fed on a balanced chow diet (not supplemented). Female rats continued to receive supplementation throughout gestation and lactation periods. The same treatment was performed for the next two generations (G1 and G2). At 75 days of age, male offspring from G1 and G2 generations from both groups were used in the experiments. G1 rats did not present any difference with control rats. However, G2 rats presented reduction in glycemia and lipidemia and improvement in in vivo insulin sensitivity (model assessment of insulin resistance, insulin tolerance test) as well as in vitro insulin sensitivity in soleus muscle (glucose uptake and metabolism). This effect was associated with increased insulin-stimulated p38 MAP kinase phosphorylation and lower n-6/n-3 fatty acid ratio, but not with activation of proteins from insulin signaling (IR, IRS-1 and Akt). Global DNA methylation was decreased in liver but not in soleus muscle. These results suggest that long-term fish oil supplementation improves insulin sensitivity in association with increased insulin-stimulated p38 activation and decreased n-6:n-3 ratio in skeletal muscle and decreased global DNA methylation in liver.     

Acute creatine administration improves mitochondrial membrane potential and protects against pentylenetetrazol-induced seizures

A growing body of evidence indicates that creatine (Cr) exerts beneficial effects on a variety of pathologies where energy metabolism and oxidative stress play an etiological role. However, the benefits of Cr treatment for epileptics are still shrouded in controversy. In the present study, we found that acute Cr treatment (300 mg/kg, p.o.) prevented the increase in electroencephalographic wave amplitude typically elicited by PTZ (30, 45 or 60 mg/kg, i.p.). Cr treatment also increased the latency periods of first myoclonic jerks, lengthened the latency periods of the generalized tonic–clonic seizures and reduced the time spent in the generalized tonic–clonic seizures induced by PTZ (60 mg/kg). Administration of PTZ (all doses) decreased Na⁺, K⁺-ATPase activity as well as adenosine triphosphate (ATP) and adenosine diphosphate levels in the cerebral cortex, but Cr treatment prevented these effects. Cr administration also prevented increases in xanthine oxidase activity, adenosine monophosphate levels, adenosine levels, inosine levels and uric acid levels that normally occur after PTZ treatment (60 mg/kg, i.p.). We also showed that Cr treatment increased the total Cr (Cr + PCr) content, creatine kinase activity and the mitochondrial membrane potential (ΔΨ) in the cerebral cortex. In addition, Cr prevented PTZ-induced mitochondrial dysfunction characterized by decreasing ΔΨ, increasing thiobarbituric acid-reactive substance levels and increasing protein carbonylation. These experimental findings reinforce the idea that mitochondrial dysfunction plays a critical role in models of epileptic seizures and suggest that buffering brain energy levels through Cr treatment may be a promising therapeutic approach for the treatment of this neurological disease.     

Metal cation toxicity in the alga Gracilaria domingensis as evaluated by the daily growth rates in synthetic seawater

Macroalgae of the genus Gracilaria have considerable economic importance as raw material for agar production and belong to an important group of organisms that are tolerant of high concentrations of metal. The median inhibitory concentration (IC₅₀) values obtained by measuring the ratio of fresh mass variation (i.e., daily growth rates) of the red macroalga Gracilaria domingensis during a 48-h aquatic toxicity assay are reported here. The alga was exposed to 14 different metal cations as well as the molybdate anion in synthetic seawater. The actual concentrations of these ionic species (at IC₅₀ values) and the proportion of free ions (aqueous complexes) were determined by inductively coupled plasma atomic emission spectroscopy and the Environmental Protection Agency-recommended software, MINTEQA2, respectively. Based on the free IC₅₀ values (IC₅₀ ^F), the ions were ranked in terms of toxicity: Cd²⁺???Cu²⁺???Pb²⁺???Zn²⁺???Ni²⁺?>?Co²⁺?>?La³⁺???Mn²⁺?>?Ca²⁺?~?Li⁺???MoO₄ ^2????Sr²⁺?>?Mg²⁺???K⁺?>?Na⁺. As a member of the first trophic level in the marine food chain, G. domingensis is an appropriate target organism both for the development of toxicological assays and as a bioindicator of marine degradation.     

Free-Running Locomotor Activity Rhythms in Cave-Dwelling Catfishes,Trichomycterus sp., from Brazil (Teleostei,Siluriformes)

Locomotor activity of troglobitic (cave restricted) catfishes, Trichomycterus sp., was recorded in the laboratory under constant darkness for ten consecutive days. Infra-red photocell beam crossings were totalled every thirty minutes and stored for later analysis. Spectral analysis of the data followed by a statistical test designed for the detection of significant components was then performed. The results of 14 individuals, seven pigmented and seven albino fishes, are reported here. In eight individuals, four pigmented and four albinos, we were not able to detect significant components in the circadian range, although ultradian and/or infradian significant components were found in all cases. Former studies on other two Brazilian cave catfishes, Pimelodella kronei and Imparfinis sp., showed similar results, suggesting a picture of gradual loss of circadian rhythmicity and persistence of ultradian and infradian rhythms in the locomotor activity of troglobitic populations. The proportion of specimens without significant circadian components was higher in Trichomycterus sp., supposedly less specialized to cave life than the other two species. This is tentatively correlated with a higher environmental stability during the time of evolutionary isolation in the subterranean biotope. These results provide strong arguments for the importance of external selection over circadian rhythmicity.     

Diurnal Variation in Oral Temperature,Sleepiness, and Performance of High School Girls

Evaluation of rhythmic fluctuations of physical and mental variables should be of special significance for the understanding of students' performance and setting the schedules of school activities. The present study investigated the pattern of diurnal variation in oral temperature, sleepiness and performance of a group of adolescents undergoing a daytime school schedule. Eighteen girls (mean age 16 years-old), who attended the same class from 0715h to 1645h, were tested on seven days. They measured their oral temperature, quantified their sleepiness level by means of a visual analogue scale, and completed the following tests: letter cancellation test, addition test, and a simple motor task. One-way ANOVA statistics for repeated measures was used in order to verify the effect of test time on oral temperature, sleepiness and performance. Possible correlations between the level of sleepiness and performance were investigated by means of Spearman rank correlation. The results revealed significant time of day effect on all variables, except for the number of addition errors. Oral temperature values showed an increase from morning to afternoon. Letter cancellation, motor task and addition scores increased from early morning to late afternoon, showing rapid fluctuations throughout the day. Sleepiness level was negatively correlated with letter cancellation scores during the first three tests of the day. In agreement with other work, the diurnal variation of oral temperature, letter cancellation and addition test showed an improvement as the day progressed. Sleepiness, on the other hand, decreased throughout the day, with the highest level associated with the first test of the day, suggesting a circadian pattern of variation rather than a cumulative effect due to school activities.