Calcium homeostasis behavior and cardiac function on left ventricular remodeling by pressure overload.

Sarcoplasmic reticulum Ca2+-ATPase (SERCA2a) and sarcolemmal Na+/Ca2+ exchanger (NCX1) structures are involved in heart cell Ca2+ homeostasis. Previous studies have shown discrepancies in their function and expression in heart failure. The goal of this study was to evaluate heart function and hypertrophied muscle Ca2+-handling protein behavior under pressure overload. Twenty male Wistar rats were divided into two groups: Aortic stenosis (AoS), induced by a clip placed at the beginning of the aorta, and Control (Sham). After 18 weeks, heart function and structure were evaluated by echocardiogram. Myocardial function was analyzed by isolated papillary muscle (IPM) at basal condition and Ca2+ protein functions were evaluated after post-pause contraction and blockage with cyclopiazonic acid in IPM. Ca2+-handling protein expression was studied by western blot (WB). Echocardiogram showed that AoS caused concentric hypertrophy with enhanced ejection fraction and diastolic dysfunction inferred by dilated left atrium and increased relative wall thickness. IPM study showed developed tension was the same in both groups. AoS showed increased stiffness revealed by enhanced resting tension, and changes in Ca2+ homeostasis shown by calcium elevation and SERCA2a blockage maneuvers. WB revealed decreased NCX1, SERCA2a, and phosphorylated phospholambam (PLB) on serine-16 in AoS. AoS had left ventricular hypertrophy and diastolic dysfunction compared to Sham; this could be related to our findings regarding calcium homeostasis behavior: deficit in NCX1, SERCA2a, and phosphorylated PLB on serine-16.

Calcium homeostasis behavior and cardiac function on left ventricular remodeling by pressure overload

Sarcoplasmic reticulum Ca2+-ATPase (SERCA2a) and sarcolemmal Na+/Ca2+ exchanger (NCX1) structures are involved in heart cell Ca2+ homeostasis. Previous studies have shown discrepancies in their function and expression in heart failure. The goal of this study was to evaluate heart function and hypertrophied muscle Ca2+-handling protein behavior under pressure overload. Twenty male Wistar rats were divided into two groups: Aortic stenosis (AoS), induced by a clip placed at the beginning of the aorta, and Control (Sham). After 18 weeks, heart function and structure were evaluated by echocardiogram. Myocardial function was analyzed by isolated papillary muscle (IPM) at basal condition and Ca2+ protein functions were evaluated after post-pause contraction and blockage with cyclopiazonic acid in IPM. Ca2+-handling protein expression was studied by western blot (WB). Echocardiogram showed that AoS caused concentric hypertrophy with enhanced ejection fraction and diastolic dysfunction inferred by dilated left atrium and increased relative wall thickness. IPM study showed developed tension was the same in both groups. AoS showed increased stiffness revealed by enhanced resting tension, and changes in Ca2+ homeostasis shown by calcium elevation and SERCA2a blockage maneuvers. WB revealed decreased NCX1, SERCA2a, and phosphorylated phospholambam (PLB) on serine-16 in AoS. AoS had left ventricular hypertrophy and diastolic dysfunction compared to Sham; this could be related to our findings regarding calcium homeostasis behavior: deficit in NCX1, SERCA2a, and phosphorylated PLB on serine-16.

Characterization of sweat induced with pilocarpine, physical exercise, and collected passively by metabolomic analysis.

BACKGROUND/PURPOSE: The elimination of the pain associated with needle picking is a strong motivation for the development of clinical non-invasive diagnostic methods. Sweat has been described as an alternative biological sample that may have a direct relation to the plasma composition. MATERIALS AND METHODS: In this study, analysis of sweat of human volunteers obtained by induction with pilocarpine is compared with sweat samples obtained by physical exercise and by passive collection along 7 hours. The sweat samples have been analyzed by 1H nuclear magnetic resonance spectroscopy. RESULTS: A range of 34 different metabolites has been detected in sweat samples, including lactate, several amino acids, pyroglutamate, and urocanate. Most of the metabolites identified were quantified. The majority of the amino acids detected in sweat seem to have origin in the epidermis surface. No significant differences in sweat samples from female and male were observed by 1H NMR metabolomic analysis. CONCLUSIONS: Principal component analysis (PCA) shows that both physical exercise and pilocarpine methods seem to be equally reproducible methods in terms of sweat metabolite composition presenting better repeatability than natural sweat collection. Nevertheless, this difference is mainly originated from amino acids with origin from the skin surface.

Lethal Fascioliasis in Capybaras ( Hydrochoerus hydrochaeris) in Brazil.

The liver fluke Fasciola hepatica (Trematoda: Fasciolidae) causes fascioliasis, which affects mostly domestic ruminants and humans worldwide. This parasite has an Old World origin and was introduced into the New World by European colonizers. Capybara ( Hydrochoerus hydrochaeris) is the largest living rodent species, with adults weighing over 60 kg. We report a fascioliasis outbreak caused by F. hepatica that reduced a capybara group from 21 to 2 animals within a 9-mo period. Animal infection and associated lesions were confirmed by postmortem examinations that revealed extensive liver damage associated with the presence of large number of adult and immature forms of F. hepatica. Both macroscopic and microscopic alterations in the liver were compatible with acute fascioliasis, which is characterized by a large parasite burden in the liver. Taxonomic identification of flukes collected from capybara livers were confirmed by molecular methods, which generated a mitochondrial NADH dehydrogenase I (NDI) gene partial sequence that was 100% identical to a F. hepatica NDI sequence from the United Kingdom. This is the first report of deleterious effects caused by F. hepatica in capybaras, highlighting the potential harm caused by this exotic parasite in the capybara.

Rosuvastatin promotes benefits on induced periodontitis in hypertensive rats.

BACKGROUND AND OBJECTIVE: The potential benefits of statins in modulating periodontal disease is supported by in vitro and clinical studies showing statins can induce a lower expression of proinflammatory cytokines and matrix metalloproteinases. The aim of this study was to evaluate the effects of rosuvastatin (ST) on ligature-induced periodontitis in spontaneously hypertensive rats (SHR). MATERIAL AND METHODS: Fifty-four adult male rats were divided into three groups: SHR-C, SHR-L and SHR-L-ST (C, control; L, ligature groups). In the SHR-L-ST group, animals were treated with daily 2 mg/kg ST administration. In L groups, a ligature remained around mandibular first molars for 10 d. Each group was divided for killing at 10 or 21 d postoperatively. Microtomographic and histometric analyses were performed. Osteoclastogenesis was evaluated by tartrate-resistant acid phosphatase assay and gene expression of 84 proinflammatory mediators by polymerase chain reaction array. RESULTS: The SHR-L-ST group showed reduced bone loss and attachment loss in comparison with the SHR-L group at both 10 and 21 d postoperatively (p < 0.05). ST decreased the amount of tartrate-resistant acid phosphatase-positive cells compared with the SHR-L group at both 10 and 21 d (p < 0.05). The SHR-L-ST group presented 14 genes differentially expressed when compared with SHR-L group, featuring a downregulated gene profile at 10 d. CONCLUSION: Statin therapy may promote a protective effect against alveolar bone and connective tissue attachment losses attributable to periodontitis in hypertensive rats through inflammatory gene profile modulation.

Effects of maternal separation on the dietary preference and behavioral satiety sequence in rats.

This study investigated the effects of maternal separation on the feeding behavior of rats. A maternal separation model was used on postnatal day 1 (PND1), forming the following groups: in the maternal separation (MS) group, pups were separated from their mothers each day from PND1 to PND14, whereas in the control (C) group pups were kept with their mothers. Subgroups were formed to study the effects of light and darkness: control with dark and light exposure, female and male (CF and CM), and maternal separation with dark and light exposure, female and male (SDF, SDM, SLF and SLM). Female rats had higher caloric intake relative to body weight compared with male controls in the dark period only (CF=23.3±0.5 v. CM=18.2±0.7, P<0.001). Macronutrient feeding preferences were observed, with male rats exhibiting higher caloric intake from a protein diet as compared with female rats (CF=4.1±0.7, n=8 v. CM=7.0±0.5, n=8, P<0.05) and satiety development was not interrupted. Female rats had a higher adrenal weight as compared with male rats independently of experimental groups and exhibited a higher concentration of serum triglycerides (n=8, P<0.001). The study indicates possible phenotypic adjustments in the structure of feeding behavior promoted by maternal separation, especially in the dark cycle. The dissociation between the mother's presence and milk intake probably induces adjustments in feeding behavior during adulthood.

Seroprevalence of Toxoplasma gondii in free-living Amazon River dolphins (Inia geoffrensis) from central Amazon, Brazil.

Toxoplasma gondii is an important pathogen in aquatic mammals and its presence in these animals may indicate the water contamination of aquatic environment by oocysts. Serum samples from 95 free-living Amazon River dolphins (Inia geoffrensis) from the Mamirauá Sustainable Development Reserve (RDSM), Tefé, Amazonas, Central Amazon, Brazil were tested for T. gondii antibodies using the modified agglutination test (MAT). Antibodies (MAT ≥ 25) to T. gondii were found in 82 (86.3%) dolphins with titers of 1:25 in 24, 1:50 in 56, and 1:500 in 2. Results suggest a high level contamination of the aquatic environment of the home range of these animals.

Molecular phylogenetic analysis in Hammondia-like organisms based on partial Hsp70 coding sequences.

The 70 kDa heat-shock protein (Hsp70) sequences are considered one of the most conserved proteins in all domains of life from Archaea to eukaryotes. Hammondia heydorni, H. hammondi, Toxoplasma gondii, Neospora hughesi and N. caninum (Hammondia-like organisms) are closely related tissue cyst-forming coccidians that belong to the subfamily Toxoplasmatinae. The phylogenetic reconstruction using cytoplasmic Hsp70 coding genes of Hammondia-like organisms revealed the genetic sequences of T. gondii, Neospora spp. and H. heydorni to possess similar levels of evolutionary distance. In addition, at least 2 distinct genetic groups could be recognized among the H. heydorni isolates. Such results are in agreement with those obtained with internal transcribed spacer-1 rDNA (ITS-1) sequences. In order to compare the nucleotide diversity among different taxonomic levels within Apicomplexa, Hsp70 coding sequences of the following apicomplexan organisms were included in this study: Cryptosporidium, Theileria, Babesia, Plasmodium and Cyclospora. Such analysis revealed the Hammondia-like organism to be the lowest divergent group when compared to other groups within the phylum Apicomplexa. In conclusion, the Hsp70 coding sequences proved to be a valuable genetic marker for phylogenetic reconstruction and may constitute a good candidate to be used with other genes for species phylogeny within this group of organisms.

Teratogenic effects of the interaction acetylsalicylic acid (ASA) and ethanol: morphologic and morphometric evaluation of the lingual epithelium in rat fetuses.

The objective of the present work was to evaluate the teratogenic effects of the interaction between acetylsalicylic acid (ASA) and ethanol on the epithelium of the lingual mucosa in rat fetuses. On the 10th pregnancy day, a single intraperitoneal ethanol dose (2.96 g/kg body weight) (Group I), ASA (200 mg/kg body weight) (Group II) and ASA plus ethanol, in the same doses (Group III), or saline (Group IV - control), were administrated. The epithelial alterations were assessed by means of histological and morphometric methods, on posterior dorsal, anterior dorsal and ventral regions of the tongue. ASA reduced, in rat fetuses, the ethanol deleterious effects on nuclear size in the epithelial prickle cell of the lingual mucosa. On the other hand, ASA did not influence the effects of ethanol in both epithelial layers of the lingual mucosa, when the nuclear shape, cell volume or epithelial layers thickness were evaluated.

Sertraline delays the somatic growth and reflex ontogeny in neonate rats.

This study investigated the somatic maturation and ontogeny of reflexes in neonate rats treated with sertraline (Sert) during the suckling period. The animals were divided into four groups; three that received daily doses of Sert (5, 10 or 15 mg/kg s.c.; groups Sert5, Sert10, and Sert15, respectively), and a fourth group that received distilled water (Dw) (1 ml/kg/b.w.). Growth indicators (body weight, axis of the head and tail length) were measured daily, from the 1st to the 21st postnatal day. The reflexes (righting, free-fall righting, negative geotaxis, cliff avoidance, auditory startle response, vibrissa placing and palm grasp) and physical-feature maturation (ear unfolding, auditory conduit opening, irruption of the lower incisors and eye opening) were recorded each day of the animal's life. All groups were compared to the Dw group. The body weight gain was reduced in all the Sert groups. Moreover, a delay in the growth of the body length was observed in all the Sert groups. Higher Sert doses reduced the speed of growth in the tail length. The medio-lateral head axis reduced in Sert15 and Sert5 doses. Otherwise, Sert10 had a temporary acceleration in this growth, but the growth of the anteroposterior head axis had a delay in all the Sert groups. The highest doses induced a delay in physical-feature maturation. The palm grasp reflex (disappearance) was retarded in Sert10; cliff avoidance advanced in Sert10; negative-geotaxis and free-fall righting retarded in Sert15. The findings suggest that altered serotonergic system activity induced by sertraline early in life could play a role in the retardation of the somatic growth ontogeny as well as a delay in the maturation of some reflexes.

Neonatal administration of citalopram delays somatic maturation in rats

We investigated the somatic maturation of neonate rats treated during the suckling period with citalopram, a selective serotonin reuptake inhibitor. Groups with 6 male neonates were randomly assigned to different treatments 24 h after birth. Each litter was suckled by one of the dams until the 21st postnatal day. Body weight, head axis and tail length were measured daily from the 1st to the 21st postnatal day. Time of ear unfolding, auditory conduit opening, incisor eruption, and eye opening was determined. Pups received 5 mg (Cit5), 10 mg (Cit10) or 20 mg/kg (Cit20) citalopram sc, or saline (0.9 percent NaCl, w/v, sc). Compared to saline, body weight was lower (24.04 percent, P < 0.01) for Cit10 from the 10th to the 21st day and for Cit20 from the 6th to the 21st day (38.19 percent, P < 0.01). Tail length was reduced in the Cit20 group (15.48 percent, P < 0.001) from the 8th to the 21st day. A reduction in mediolateral head axis (10.53 percent, P < 0.05) was observed from the 11th to the 21st day in Cit10 and from the 6th to the 21st day in Cit20 (13.16 percent, P < 0.001). A reduction in anteroposterior head axis was also observed in the Cit20 group (5.28 percent, P < 0.05) from the 13th to the 21stday. Conversely, this axis showed accelerated growth from the 12th to the 21stday in the Cit5 group (13.05 percent, P < 0.05). Auditory conduit opening was delayed in the Cit5 and Cit20 groups and incisor eruption was delayed in all citalopram groups. These findings show that citalopram injected during suckling to rats induces body alterations and suggest that the activity of the serotoninergic system participates in growth mechanisms.

Neonatal administration of citalopram delays somatic maturation in rats.

We investigated the somatic maturation of neonate rats treated during the suckling period with citalopram, a selective serotonin reuptake inhibitor. Groups with 6 male neonates were randomly assigned to different treatments 24 h after birth. Each litter was suckled by one of the dams until the 21st postnatal day. Body weight, head axis and tail length were measured daily from the 1st to the 21st postnatal day. Time of ear unfolding, auditory conduit opening, incisor eruption, and eye opening was determined. Pups received 5 mg (Cit5), 10 mg (Cit10) or 20 mg/kg (Cit20) citalopram sc, or saline (0.9% NaCl, w/v, sc). Compared to saline, body weight was lower (24.04%, P < 0.01) for Cit10 from the 10th to the 21st day and for Cit20 from the 6th to the 21st day (38.19%, P < 0.01). Tail length was reduced in the Cit20 group (15.48%, P < 0.001) from the 8th to the 21st day. A reduction in mediolateral head axis (10.53%, P < 0.05) was observed from the 11th to the 21st day in Cit10 and from the 6th to the 21st day in Cit20 (13.16%, P < 0.001). A reduction in anteroposterior head axis was also observed in the Cit20 group (5.28%, P < 0.05) from the 13th to the 21st day. Conversely, this axis showed accelerated growth from the 12th to the 21st day in the Cit5 group (13.05%, P < 0.05). Auditory conduit opening was delayed in the Cit5 and Cit20 groups and incisor eruption was delayed in all citalopram groups. These findings show that citalopram injected during suckling to rats induces body alterations and suggest that the activity of the serotoninergic system participates in growth mechanisms.

Prevalence of Neospora caninum antibodies and factors associated with their presence in dairy cattle of the north of Paraná state, Brazil.

To determine the prevalence of Neospora caninum antibodies and associated factors, blood sera from 623 female dairy cattle from 23 farms in the north of the state of Paraná, Brazil, were analyzed by means of the indirect immunofluorescent-antibody test (IFAT > or = 25). Serum samples from 134 dogs living on the same farms also were tested for N. caninum antibodies (IFAT > or = 50), and the presence of dogs was associated with the prevalence of N. caninum antibodies in cattle. The overall seroprevalence in cattle was 14.3%, mainly in animals over 24 months of age. Seroprevalence in Holsteins (15.1% of 558) was greater than in mixed-breed cattle (7.7% of 65). Age (> or =24 months) of cattle, feeding silage and/or concentrate produced on the farm were associated; antibodies were found in 21.6% of dogs; and the presence of dogs was associated with the prevalence of N. caninum antibodies in cattle.

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in ovine from São Paulo State, Brazil.

Serum samples from 597 sheep from São Paulo State, in the southeastern region of Brazil, were tested to determine the prevalence of antibodies directed against Toxoplasma gondii (> or = 1:64) and Neospora caninum (> or = 1:50) using the indirect fluorescent antibody test (IFAT). The animals were divided into three groups based on their age: < or = 1 year, 1-4 years, and > or = 4 years. Antibodies to T. gondii were observed in 34.7% of the samples with titers ranging from 64 to 16,384 and IgG antibodies directed against N. caninum were observed in 9.2%, with titers ranging from 50 to 3200. Only 3.5% of the sheep were positive for both agents. All farms had at least one positive animal for T. gondii, and 26 of the 30 farms had at least one positive animal for N. caninum. An association between seroprevalence and age was observed for T. gondii (P = 0.001), but not to N. caninum (P = 0.343). It was not possible to associate seroprevalence to T. gondii and the presence of domestic or feral cats, since in all farms there was at least one positive sheep. There was no association between seropositivity to N. caninum and the presence of domestic (P = 1.000) and feral dogs (P = 0.550).

Detection of antibodies to Neospora caninum in two species of wild canids, Lycalopex gymnocercus and Cerdocyon thous from Brazil.

Domestic dog (Canis domesticus) and the coyote (Canis latrans) are the only known definitive hosts for the protozoan Neospora caninum that causes abortion in dairy cattle. In the present study, antibodies to N. caninum were sought in three species of wild canids, Cerdocyon thous, Lycalopex gymnocercus and Dusicyon vetulus from Brazil. Antibodies to N. caninum were assayed by the indirect fluorescent antibody test (IFAT) and the Neospora agglutination test (NAT). N. caninum antibodies were found in five of 12 L. gymnocercus with IFAT titers of 1:50 in three, 1:100 in one, and 1:1600 in one, and NAT titers of 1:40, 1:80, 1:160, 1:320, and 1:640 in five animals. Antibodies to N. caninum were found in four of 15 C. thous with IFAT titers of 1:50 in one, and 1:100 in three, and NAT titer of 1:40 in one animal. All 30 D. ventulus were seronegative by IFAT and NAT.

Prevalence of anti-Neospora caninum antibodies in dogs from urban, periurban and rural areas of the city of Uberlândia, Minas Gerais--Brazil.

In Brazil there are few reports on the prevalence of anti-Neospora caninum antibodies in dogs from urban, periurban and rural areas. Serum samples from 450 dogs, 300 from urban, 58 from periurban and 92 from rural areas, were tested for the detection of anti-N. caninum IgG antibodies using IFAT: indirect fluorescent antibody test (IFAT, > or =50). Antibodies were observed in 63 (14%) of the 450 serum samples analyzed, with 32 (10.7%) in dogs coming from the urban area, 11 (18.9%) from the periurban area and 20 (21.7%) from the rural area. Statistical differences were seen between the occurrences in animals from the urban area and those of the rural area (P = 0.01). The antibody titers found were: 1:50 in 20 dogs, 1:100-1:800 in 41 dogs, and 1:3200 in two dogs. In the female dogs, a smaller prevalence of dogs with antibodies was observed in those from the urban area (7.5%) in comparison with those of the rural (21.0%) (P = 0.05) and periurban (23.3%) (P = 0.01) areas. There were growing levels of antibody prevalence with the increase in age of the dogs in all three areas studied. Although this increase was not significant, it indicates a tendency towards more infections with age, suggesting post-natal exposure to N. caninum. However, a significant difference (P = 0.05) was observed in the occurrence of anti-N. caninum antibodies in dogs with ages = 2 years in urban (13.1% urban) versus rural environments (27.1% rural). Among the other age brackets studied the difference was not significant. The results confirm the presence of N. caninum in the region and reveal the important role of dogs in the parasite's epidemiology.

Prevalence of antibodies to Neospora caninum in dogs from Amazon, Brazil.

Neospora caninum is an important cause of abortion in dairy cattle worldwide. Dogs are important in the epidemiology of this parasite because they are the only hosts known to excrete N. caninum oocysts. Antibodies to N. caninum were assayed in serum samples from 157 dogs from Monte Negro, Rondônia, Amazon, Brazil using the indirect fluorescent antibody test. Antibodies to N. caninum were found in 13 (8.3%) of dogs in titers of 1:50 in 1, 1:100 in 2, 1:200 in 5, 1:800 in 1, 1:1600 in 2, and 1:3200 in 2 dogs. These data indicate that N. caninum infection is prevalent even in remote areas of the Amazon.

Early malnourished rats are not affected by anorexia induced by a selective serotonin reuptake inhibitor in adult life.

The effect of early postnatal malnutrition upon food intake and its modulation by the selective serotonin reuptake inhibitor (SSRI) citalopram, was investigated in adult rats. Sixty four Wistar rats were allocated to two groups, according to their mother's diet during lactation. Mothers receiving a 23% protein diet fed the well-nourished group; mothers receiving 8% protein diet fed the malnourished. After weaning, all rats received the 23% protein diet ad libitum. On the 120th day after birth, each nutritional group was divided in two subgroups (each one, n = 16) which received a single daily injection of citalopram (10 mg/kg) or saline (0.9% NaCl) for 14 days. Chronic treatment with citalopram decreased both the food intake and weight gain in the well-nourished rats, but not in the malnourished ones. These data are consistent with findings concerning the nutritional manipulation of the nervous system during its higher vulnerable phase, suggesting that early malnutrition alters the effect of treatment of SSRI in adult rats, and that malnutrition during the critical period of brain development affects the serotoninergic system.

Prevalence of Neospora caninum antibodies in dogs from dairy cattle farms in Parana, Brazil.

Serum samples from 134 dogs from 22 cattle dairy farms in the northern region of Parana State, Brazil, were tested for antibodies to Neospora caninum in an indirect fluorescent antibody test. Antibodies (> or = 1:50) to N. caninum were found in 29 (21.6%) of the 134 dogs, and seropositive dogs were found on 14 (63.6%) of the 22 dairy cattle farrms. The antibody titers of dogs were 1:50 (3 dogs), 1:100 (7 dogs), 1:200 (7 dogs), 1:400 (6 dogs), and > or = 1:800 (6 dogs). The low prevalence (9%) in < 1-yr-old dogs compared with the 2- to 3-fold higher prevalence in older dogs (17-29%) suggests postnatal exposure to N. caninum infection.