Background & objectives: In Portugal, Phlebotomus perniciosus and P. ariasi,(Subgenus Larroussius; Diptera: Psychodidae) are the proven vectors of leishmaniasis caused by Leishmania infantum. The Algarve Region in southern Portugal has been considered an endemic focus of leishmaniasis since 1980s. The main objective of the present study was to validate a molecular approach to detect Leishmania infection in phlebotomines based on DNA extraction from the female sandfly whole body, minus genitalia, followed by PCR for application on epidemiological surveys. Methods: In Algarve Region, from early May until early November 2006, sandflies were captured by CDC miniature light-traps. kDNA-PCR and ITS1-PCR were used to screen the presence of Leishmania DNA in female sandflies after species identification by entomological keys. Results: A total of 474 sandflies were collected in 108 biotopes. One female of P. perniciosus, the predominant species, was found infected with L. infantum reflecting an overall infection rate of 0.47%. Interpretation & conclusion: PCR associated with morphological characterization of the sandflies will be a powerful epidemiological tool for the determination of the number of phlebotomines infected with Leishmania spp in nature. In addition, the simultaneous occurrence of dogs and P. perniciosus infected with L. infantum shows that Algarve continues to be an endemic focus of canine leishmaniasis. Furthermore, as P. sergenti and P. papatasi which transmit L. tropica and L. major, respectively were present, the future introduction of these two Leishmania species in southern region of Portugal should not be neglected.

BACKGROUND:/objective. Basic science advances in spinal cord injury (SCI) are leading to novel clinical approaches.The authors report a prospective, uncontrolled pilot study of the safety and outcomes of implanting olfactory mucosal autografts (OMA) in 20 patients with chronic,sensorimotor complete or motor complete SCI. METHODS:.Seven paraplegic and 13 tetraplegic subjects (17 men and 3 women; 19-37 years old) who sustained a traumatic SCI 18 to 189 months previously (mean = 49 months) were enrolled. Preoperative rehabilitation that emphasized lower extremity stepping using either overground walking training or a robotic weight-supported treadmill training was provided for 25 to 39 hours per week for a median of 4 months at 3 sites. No change in ASIA Impairment Scale (AIS) motor scores for the lower extremities or AIS grades of completeness was found. OMAs were transplanted into 1.3-to 4-cm lesions at C4-T12 neurological levels after partial scar removal.Therapy was continued postoperatively.Preoperative and postoperative assessments includedAIS scores and classification, electromyography (EMG) of attempted voluntary contractions, somatosensory evoked potentials (SSEP),urodynamic studies with sphincter EMG, spinal cord magnetic resonance imaging (MRI), and otolaryngology and psychology evaluations.The Functional Independence Measure (FIM) and Walking Index for Spinal Cord Injury (WISCI) were obtained in 13 patients. RESULTS:. All patients survived and recovered olfaction.One patient was rehospitalized for aseptic meningitis.Minor adverse events occurred in 4 others.The mean duration of follow-up was 27.7 months (range = 12-45 months). By MRI, the lesion site was filled in all patients with no neoplastic overgrowth or syringomyelia.AIS grades improved in 11 of 20 patients, 6 (A --> C),3 (B --> C),and 2 (A --> B), and declined in 1 (B --> A). Improvements included new voluntary EMG responses (15 patients) and SSEPs (4 patients). Scores improved in the FIM and WISCI (13/13 tested), and urodynamic responses improved in 5 patients. CONCLUSION:. OMA is feasible, relatively safe, and possibly beneficial in people with chronic SCI when combined with postoperative rehabilitation. Future controlled trials may need to include a lengthy and intensive rehabilitation arm as a control.

Pregnant rats were exposed to ethanol (EtOH) and/or methyl mercury (MeHg) during fetal brain development. Nitrergic activity was quantified by densitometric measurement of formazan deposits in the hippocampus, cerebellum and striatum of two-month-old offspring following histochemical assay for NADPH-diaphorase (NADPH-d) activity. Compared to control subjects, an increase in nitrergic activity was found in the molecular layer of dentate gyrus and in the lacunosum molecular and stratum radiatum of CA1 (cornus amoni 1) in the EtOH+MeHg group, whereas a single administration of EtOH increased the activity in all striatal segments. The cerebellum seems to be less sensitive at this time-point to intoxication, and presented an increase only at the molecular layer of EtOH-exposed animals when compared to the MeHg and EtOH+MeHg groups (ANOVA, one-way followed by Tukey's test, p<0.05 or p<0.01). Taken together, results suggest that developmental exposure to EtOH and MeHg, singularly or in combination, alters nitrergic activity in adult rat in different ways depending on the region and layer of the central nervous system (CNS), and that these alterations might be related to different local metabolic properties.

Regucalcin plays an important role in maintenance of intracellular Ca(2+) homeostasis, suppresses cell proliferation, inhibits expression of oncogenes, and increases the expression of tumour suppressor genes. This suggests that regucalcin functions may be altered in cancer tissues. In this study the regucalcin expression in breast and prostate cancer cases was analysed by RT-PCR and immunohistochemistry showing that the mRNA and/or protein are under-expressed in these tumors. The effect of sex steroid hormones on regucalcin expression in breast and prostate cancer cells was determined by real-time PCR. MCF-7 and LNCaP cells were stimulated with 0, 1, and 10 nM of 17beta-estradiol (E(2)) or 5alpha-dihydrotestosterone (DHT), respectively, for 0, 6, 12, 24, and 48 h. MCF-7 cells were also stimulated with E(2) conjugated to BSA (E(2)-BSA). To explore the mechanisms underlying the sex steroid regulation of regucalcin expression, control treatments with ICI 182,780, flutamide and cyclohexamide were carried out. E(2) effects regulating regucalcin expression were not abrogated in the presence of ICI 182,780, and were similar to those observed with E(2)-BSA, which suggests the involvement of a membrane-bound estrogen receptor. In LNCaP cells, DHT down-regulated regucalcin expression, an effect inhibited by the presence of both flutamide and cyclohexamide, suggesting the involvement of androgen receptor and de novo protein synthesis. The loss of regucalcin expression in breast and prostate cancer cases and the regulation of its expression by sex steroid hormones suggest that it may be associated with development and progression of these human tumors. J. Cell. Biochem.(c) 2009 Wiley-Liss, Inc.

Studies involving alcohol and its interactions with other neurotoxicants represent the focus of several works of research due to the fact that the use of alcohol can sometimes leads to serious health problems. Fetal exposure to alcohol and mercury has a high incidence in some regions of Brazil, where there are pregnant women who are alcoholics and live in mining areas. This work was conducted to examine the effects of combined exposure to ethanol (EtOH) and methylmercury (MeHg) in rats during the development of the central nervous system (CNS). Experimental behavioral animal models/tests were used in order to examine locomotion, anxiety, depression and memory. Pregnant rats received tap water or EtOH 22.5%(w/v)(6.5g/(kgday), by gavage) during pregnancy and breast-feeding. On the 15th day of pregnancy, some groups received 8mg/kg of MeHg (by gavage). The groups were as follows: control, EtOH, MeHg and EtOH+MeHg. The experimental results showed that the EtOH, MeHg and EtOH+MeHg groups reduced the percentage of frequency and time spent in the open arms entries of the elevated plus-maze (EPM) test, when compared to the control group. This result suggests an anxiogenic behavioral response. The MeHg group increased locomotor activity in the arena and the immobility time in the forced swimming test, suggestive of depression-like behavior. The EtOH+MeHg group showed greater reductions in the percentages of frequency and time spent in the open arms entries in the EPM test, suggesting a sedative-behavior since the frequency of enclosed arm entries was affected. In the inhibitory avoidance task, the EtOH+MeHg group reduced the latency of the step-down response onto the grid floor, suggesting a cognitive and behavior dysfunctions. Taken together, the results suggest that EtOH and/or MeHg intoxication during the developing CNS may be a risk for deficits related to locomotor impairment, anxiety, depression and neurocognitive functions. There is a possibility that EtOH may prevent some of the MeHg responses, but the precise mechanism of action involved in this process needs to be considered for future research.

Six transmembrane epithelial antigen of the prostate 1 (STEAP1) was identified as a prostate-specific cell-surface antigen over-expressed in prostate cancer, and in human cancer cell lines obtained from several other tissues. Its cell surface location in all tumor types analyzed so far, and its absence in most vital organs in humans, turned STEAP1 into a potential target for anti-tumor immunotherapy. This study provides experimental evidence that STEAP1 is also over-expressed in human breast cancer cases, and in normal breast tissue adjacent to breast tumors, where it is localized in the cell membrane of epithelial cells. It is also demonstrated that STEAP1 transcription correlates negatively with estrogen receptor (ER) immunoreactivity, and positively with tumor grading in breast cancer cases. As estrogens are involved in breast cancer onset and progression, the response of STEAP1 to 17beta-estradiol (E(2)) was investigated in the mammary gland of rats, and in the human breast cancer cell line, MCF-7. These experiments demonstrated that STEAP1 is down-regulated by E(2) in both models. The mechanisms underlying the STEAP1 response to E(2) in vitro were further investigated in MCF-7 cells, and the results obtained suggest an effect mediated by the membrane-bound ERalpha (mbERalpha).

Canine leishmaniasis (CanL) caused by Leishmania infantum (syn. L. chagasi, in Latin America), which is transmitted by the bite of phlebotomine sand flies, is endemic and affects millions of dogs in Europe, Asia, North Africa and South America. It is an emergent disease in North America. Early detection and treatment of infected animals may be critical in controlling the spread of the disease and is an essential part of human zoonotic visceral leishmaniasis control. The laboratory diagnosis of CanL still poses a challenge, despite progress made in the development of several direct and indirect methods. An effective diagnosis test, apart of being able to confirm a clinical suspicion in a single patient as well as to detect infection in asymptomatic dogs, should have high sensitivity, specificity and reproducibility; it must be simple, easy to perform, non-expensive, feasible in regional laboratories or adaptable for field conditions. Ideally, it should detect all Leishmania-infected dogs, preferentially using non-invasive collection of biological samples. In this paper we review the advantages and shortcomings of the available procedures for CanL diagnosis in the different phases, e.g. pre-patent and patent period of the infection and methods to determine the related immune response.

Conformational changes in the structures of humic acids (HA) extracted from compost with varying degrees of maturity were monitored by high performance size exclusion chromatography (HPSEC). The molecular size distribution of HA was compared in solutions containing sodium or ammonium counterions at pH 7 and pH 4.5. These findings indicate that the humates' molecular size depended not only on the nature of the counterions but also on their concentration in the solution. The physicochemical nature of sodium counterions determined smaller molecular sizes than those of the more hydrated ammonium counterions, at low concentrations of humates. Conversely, at higher humate concentrations, the more compact conformation of sodium humates produced larger molecular sizes than those of ammonium humates due to the aggregation of more hydrophobic surfaces in the sodium humates. Composting led to the degradation of labile microbial components with accumulation of hydrophobic constituents. This caused self-association of hydrophobic compounds into humic superstructures of larger molecular size over composting time. At lower pH, changes in conformational stability by the addition of acetic acid to humate solutions were explained by the supramolecular model of humified organic matter.

Leishmaniasis, caused by Leishmania infantum, is an endemic zoonosis in the Mediterranean basin. Dogs are considered the major host for these parasites, as well as the main reservoir for human visceral infection. In recent years, asymptomatic infection or clinical disease caused by L. infantum in cats has been reported in several countries where zoonotic leishmaniasis is present. The aim of the present study was to perform a leishmaniasis survey in cats from an endemic focus. Twenty-three adult stray cats were surveyed by clinical examination, and peripheral blood samples for serological and molecular analysis were collected. In 7 of the 23 cats (30.4%) Leishmania DNA was detected in blood. A low level of fluorescent antibodies was detected in four serum samples. All the animals were asymptomatic. Taking into account the high rate of asymptomatic feline leishmaniasis in this survey, it can be suggested that cats may act as a habitual reservoir host of L. infantum infection in endemic areas. Furthermore, it will be important in the future to add this parasitosis to the differential diagnosis of feline infections from leishmaniasis foci in cats. Feline leishmaniasis diagnosis should be accessed by molecular tools.

The protozoan Giardia lamblia is the most frequent intestinal parasite of first-world countries and a major cause of waterborne disorder often referred to as traveler's diarrhea. We have previously noticed that the putrescine analog 1,4-diamino-2-butanone (DAB) remarkably inhibits the growth of anaerobic trichomonad and Trypanosoma cruzi parasites. Here, we examined the role of polyamines in Giardia cells using this putrescine analog. DAB impaired parasite proliferation dose-dependently. The analog induced increased flagella numbers and sometimes four ventral disks as well as asymmetrical division, indicating truncated or deregulated cytokinesis. Electron microscopy analysis revealed that DAB also triggered the encystment process. Oxidative stress was evaluated by measuring lipid peroxidation by thiobarbituric acid reactive substances (TBARS) detection. Trophozoites incubated either with 1 mM of DAB or putrescine for 18 h displayed increased lipoperoxide levels. Addition of 200 muM aminoguanidine, a polyamine/diamine oxidase inhibitor, partially reverted the DAB, but not the putrescine effects, indicating that the DAB effects are due, at least in part, to DAB oxidation end products. These data indicate that polyamines play a role in Giardia cell division, differentiation, and antioxidant defenses.