Clinical, hematological and biochemical alterations in hamster (Mesocricetus auratus) experimentally infected with Leishmania infantum through different routes of inoculation.

BACKGROUND: Leishmaniasis remains among the most important parasitic diseases in the developing world and visceral leishmaniasis (VL) is the most fatal. The hamster Mesocricetus auratus is a susceptible model for the characterization of the disease, since infection of hamsters with L. infantum reproduces the clinical and pathological features of human VL. In this context, it provides a unique opportunity to study VL in its active form. The main goal of this study was to evaluate the clinical, biochemical, and hematological changes in male hamsters infected through different routes and strains of L. infantum. METHODS: In the current study, hamsters (Mesocricetus auratus) were infected with the L. infantum strains (WHO/MHOM/BR/74/PP75 and MCAN/BR/2008/OP46) by intradermal, intraperitoneal and intracardiac routes. The animals were monitored for a nine month follow-up period. RESULTS: The hamsters showed clinical signs similar to those observed in classical canine and human symptomatic VL, including splenomegaly, severe weight loss, anemia, and leucopenia. Therefore the OP46 strain was more infective, clinical signs were more frequent and more exacerbated in IC group with 80 to 100 % of the animals showing splenomegaly, in the last month infection. Additionally, desquamation, hair loss and external mucocutaneous lesions and ulcers localized in the snout, accompanied by swelling of the paws in all animals, were observed. Consequently, the animals presented severe weight loss/cachexia, hunched posture, an inability to eat or drink, and non-responsiveness to external stimuli. Furthermore, regardless of strain, route of inoculum and time assessed, the animals showed renal and hepatic alterations, with increased serum levels of urea and creatinine as well as elevated serum levels of aspartate aminotransferase and alanine aminotransferase. CONCLUSIONS: These results strongly suggest that the inoculation through the intracardiac route resulted in a higher severity among infections, especially in the sixth and ninth month after infection via intracardiac, exhibited clinical manifestations and biochemical/hematological findings similar to human visceral leishmaniasis. Therefore, we suggest that this route must be preferentially used in experimental infections for pathogenesis studies of VL in the hamster model.

Temperature effects on the fracture resistance of scales from Cyprinus carpio.

In this investigation the fracture resistance of scales from Cyprinus carpio was evaluated as a function of environmental temperature. Tear specimens were prepared from scales obtained from three characteristic regions (i.e. head, mid-length and tail) of multiple fish. The fracture resistance was characterized in Mode III loading and over temperatures ranging from -150°C to 21°C. Results showed that there was a significant reduction in tear resistance with decreasing temperature and the lowest resistance to fracture was obtained at -150°C. There was a significant difference in the relative tear toughness between scales from the three locations at ambient conditions (21°C), but not below freezing. Scales obtained near the head exhibited the largest resistance to fracture (energy ≈ 150 ± 25 kJm(-2)) overall. The fracture resistance was found to be primarily dependent on the thickness of the external mineralized layer and the number of external elasmodine plies, indicating that both the anatomical position and the corresponding microstructure are important to the mechanical behavior of elasmoid fish scales. These variables may be exploited in the design of bioinspired armors and should be considered in future studies concerning the mechanical behavior of these interesting natural materials.

Ras1CA-upregulated BCPI inhibits cathepsin activity to prevent tissue destruction of the Bombyx posterior silk gland.

Using the GAL4/UAS transgenic system established in the silkworm, Bombyx mori, we have previously reported that overexpression of the Ras1(CA) oncogene specifically in the posterior silk gland (PSG) resulted in improved fibroin synthesis, silk yield, and other phenotypic effects. However, the detailed molecular mechanism remains to be fully elucidated. Using 2D-DIGE-MS/MS analyses, we compared the proteomic profiles of PSGs from the wild type (WT) and Ras1(CA)-overexpressed silkworms. Among the 24 Ras1(CA)-enhanced proteins, the Bombyx cysteine protease inhibitor (BCPI) was increased 2.4-fold at the protein level and 3.4-fold at the mRNA level. Consistent with the developmental profiles, injection of recombinant BCPI into the WT silkworms at the early wandering stage inhibited cathepsin activity, prevented tissue destruction of the PSG, and delayed pupation. Moreover, injection of small-molecule inhibitors of cathepsin into the WT silkworms prevented PSG destruction and delayed pupation, confirming the role of BCPI in inhibiting cathepsin activity. Furthermore, injection of chemical inhibitors of the Ras downstream effectors into the Ras1(CA)-overexpressed and WT silkworms revealed that both Raf-MAPK and PI3K-TORC1 pathways were required for Ras1 to induce bcpi expression. Taken together, we conclude that via the downstream Raf-MAPK and PI3K-TORC1 pathways, Ras1(CA) upregulates bcpi, which inhibits cathepsin activity thus preventing PSG destruction in Bombyx.

Tyrosine hydroxylase immunoreactivity as indicator of sympathetic activity: simultaneous evaluation in different tissues of hypertensive rats.

Vasomotor control by the sympathetic nervous system presents substantial heterogeneity within different tissues, providing appropriate homeostatic responses to maintain basal/stimulated cardiovascular function both at normal and pathological conditions. The availability of a reproducible technique for simultaneous measurement of sympathetic drive to different tissues is of great interest to uncover regional patterns of sympathetic nerve activity (SNA). We propose the association of tyrosine hydroxylase immunoreactivity (THir) with image analysis to quantify norepinephrine (NE) content within nerve terminals in arteries/arterioles as a good index for regional sympathetic outflow. THir was measured in fixed arterioles of kidney, heart, and skeletal muscle of Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) (123 ± 2 and 181 ± 4 mmHg, 300 ± 8 and 352 ± 8 beats/min, respectively). There was a differential THir distribution in both groups: higher THir was observed in the kidney and skeletal muscle (∼3-4-fold vs. heart arterioles) of WKY; in SHR, THir was increased in the kidney and heart (2.4- and 5.3-fold vs. WKY, respectively) with no change in the skeletal muscle arterioles. Observed THir changes were confirmed by either: 1) determination of NE content (high-performance liquid chromatography) in fresh tissues (SHR vs. WKY): +34% and +17% in kidney and heart, respectively, with no change in the skeletal muscle; 2) direct recording of renal (RSNA) and lumbar SNA (LSNA) in anesthetized rats, showing increased RSNA but unchanged LSNA in SHR vs. WKY. THir in skeletal muscle arterioles, NE content in femoral artery, and LSNA were simultaneously reduced by exercise training in the WKY group. Results indicate that THir is a valuable technique to simultaneously evaluate regional patterns of sympathetic activity.

Effects of shell lesions on survival, growth, condition and reproduction in the New Zealand blackfoot abalone Haliotis iris.

The pathogenicity of shell lesions in Haliotis iris Martyn was examined in a laboratory experiment in which 73 apparently healthy and 106 lesion-bearing abalone were maintained for up to 12 mo. The abalone were collected from the wild and kept in cages (1 ind. cage(-1)) for 4, 8 or 12 mo, at which times estimates of survival, growth, condition and reproductive capacity were made for each of 3 groups: 'healthy' (n = 73), 'mildly affected' (n = 61) and 'severely affected' (n = 32). Unaffected abalone showed a 2.7% mortality (n = 73) compared to 7.5% (n = 93) in lesion-bearing individuals over the entire experiment. Growth rates were significantly decreased in mildly and severely affected abalone: the relative von Bertalanffy growth coefficient (K), calculated over 12 mo, was -0.176 for unaffected, -0.079 for mildly affected and -0.048 for severely affected individuals. The asymptotic length (L(infinity)) was calculated to be 131.5 mm for unaffected, 142.1 mm for mildly affected and 150.3 mm for severely affected abalone. Significantly (p < 0.05) lower condition indices and decreased reproductive capacity (p > 0.05) were obtained for the severely affected group compared to unaffected abalone. These trends were consistent over the course of the experiment.