Could polymorphisms in ABCB1 gene represent a genetic risk factor for the development of mammary tumors in dogs?

The ABCB1 gene encodes the P-glycoprotein (P-gp) which regulates distribution and bioavailability of many endogenous and exogenous substrates, acting as a cellular mechanism of protection against these substances. Some studies have shown evidence that P-gp is related to carcinogenesis. In this study, we performed PCR and direct sequencing of ABCB1 exons 9 and 26 in 47 tissue DNA samples from canine mammary tumors. A statistically significant correlation between distinct canine breeds and the frequency of ABCB1 polymorphisms (c.985T > A and c.3442A > G SNP in ABCB1exons 9 and 26, respectively) was observed (P = 0.0015). In contrast, the TNM clinical staging, age, histological type and grade, as well as other histopathological characteristics, did not present statistically significant difference in relation to one or both SNP found in exons 9 and 26. These findings raise questions about the role of the canine ABCB1 polymorphisms in the development of mammary tumors, since the Poodle breed, which is the most common dog breed affected by mammary tumors in Brazil, presented the highest frequency of these variants. Notwithstanding, additional studies comprising a number of samples expressing the ABCB1 gene from healthy dogs, with advanced age and from different breeds, will be necessary to confirm the association of ABCB1polymorphisms and the development of mammary tumors.

Common germline haplotypes and genotypes identified in BRCA2 exon 11 of dogs with mammary tumours and histopathological analyses.

The canine BRCA2 is a tumor supressor gene which encodes the BRCA2 protein, involved in DNA repair through interaction with the RAD51 recombinase. This process is mediated by eigth BRC repeats that are encoded by BRCA2 exon 11. Two variants corresponding to human mutations in human BRC3 repeat have been reported in canine BRC3 repeat. In addition, other variants have also been described in canine BRCA2 exon 11. Considering the importance of polymorphisms in human BRCA2 to breast cancer development, this study aimed to investigate the frequency of variants in BRCA2 exon 11 in 48 blood and tissue DNA samples from bitches with canine mammary tumors (CMT), as well as, to analyze tumor stage and histopathological features. Seven Single Nucleotide Polymorphisms (SNPs) were identified, three of which were evaluated as possibily or probably deleterious variant. Interestingly, almost all the 22 mammary tumors (except one) which presented a clinical staging equal to or greater than III carried at least one mutant allele of these three variants. Besides that, no statistically significant correlation was observed between any of the reported SNPs in heterozygosis or homozygosis and either dogs data (such as breed, age or disease stage) or mammary tumors histopathological characteristics. A total of 97.9% of bitches had one to three polymorphisms of the seven identified in this study, which suggests a possibly correlation between the canine BRCA2 exon 11 polymorphisms and mammary carcinogenesis.

Rickettsia parkeri in free-ranging wild canids from Brazilian Pampa.

Spotted fevers are tick-borne diseases associated with various Rickettsia species. Rickettsia parkeri sensu stricto (s.s.) is the agent of an emerging eschar-associated rickettsiosis in humans from the USA and South American Pampa. Considering that R. parkeri s.s. is restricted to Americas and the potential role of dogs in the epidemiology of the disease, it is thus reasonable to hypothesize that wild canids could be involved in the enzootic cycle of this rickettsiosis. The aim of this work was to investigate the potential role of the wild canids from Pampa, Cerdocyon thous (crab-eating fox) and Lycalopex gymnocercus (Pampas fox), in the ecology of R. parkeri s.s. For that, 32 live-trapped free-ranging wild canids were sampled. Ticks were observed in 30 of the 32 foxes. Of the 292 ticks collected, 22 (7.5%) were positive by PCR for the presence of R. parkeri s.s. DNA. Also, 20 (62%) wild canids showed antibodies against R. parkeri. The results suggest that wild canids are involved in the enzootic cycle of R. parkeri s.s. in the Pampa biome and could be responsible for pathogen (and its vectors) dispersal.

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.

Vectorial competence of Amblyomma tonelliae to transmit Rickettsia rickettsii.

The aim of this work was to test the vectorial competence of Amblyomma tonelliae (Ixodida: Ixodidae) to transmit Rickettsia rickettsii (Rickettsiales: Rickettsiaceae), the agent of Rocky Mountain spotted fever (RMSF). All parasitic stages of A. tonelliae were exposed to R. rickettsii by allowing each stage to feed on hosts inoculated with this pathogen. Thereafter, ticks were fed on uninfected hosts. All stages of A. tonelliae were able to acquire the R. rickettsii infection and maintain it by transstadial and transovarial transmission. When infected ticks fed on uninfected hosts, the hosts developed rickettsiosis disease. This study demonstrates the vectorial competence of A. tonelliae to transmit R. rickettsii. These results have epidemiological relevance because A. tonelliae is one of the tick species most likely to infest humans in Argentina, including in areas in which RMSF has been reported.

Dynamics of Exposure to Rickettsia parkeri in Cattle in the Paraná River Delta, Argentina.

Several cases of human rickettsiosis caused by Rickettsia parkeri were recently documented in the Paraná River delta of Argentina, where the tick vector is Amblyomma triste Koch. As cattle suffer recurrent A. triste infestations, they are at risk of becoming infected with R. parkeri Herein we investigated the dynamics of R. parkeri and its A. triste vector in a herd of beef cattle. Cattle were followed for 18 mo and samples were analyzed for the presence of antibodies against four Rickettsia species (R. parkeri, Rickettsia bellii, Rickettsia amblyommii, and Rickettsia felis) and also for the presence of rickettsial DNA. Additionally, cattle were examined for attached ticks and questing adult ticks were collected. All ticks were analyzed for the presence of rickettsial DNA. No evidence of rickettsemia was found in any cow, but the high R. parkeri infection rate documented in A. triste both questing in the study area (13.9%) and feeding on cattle (19.8%) and the identification of antibodies against R. parkeri antigen in 90% of cattle are evidence that infection is taking place. Altogether, our data suggest that A. triste ticks are capable of naturally exposing cattle to R. parkeri However, the progress of R. parkeri infection and its impact on bovine health and production remain to be established.

The potential of secondary metabolites from plants as drugs or leads against protozoan neglected diseases - part II.

Infections with protozoan parasites are a major cause of disease and mortality in many tropical countries of the world. Diseases caused by species of the genera Trypanosoma (Human African Trypanosomiasis and Chagas Disease) and Leishmania (various forms of Leishmaniasis) are among the seventeen "Neglected Tropical Diseases" (NTDs) defined by the WHO. Furthermore, malaria (caused by various Plasmodium species) can be considered a neglected disease in certain countries and with regard to availability and affordability of the antimalarials. Living organisms, especially plants, provide an innumerable number of molecules with potential for the treatment of many serious diseases. The current review attempts to give an overview on the potential of such plant-derived natural products as antiprotozoal leads and/or drugs in the fight against NTDs. In part I, a general description of the diseases, the current state of therapy and need for new therapeuticals, assay methods and strategies applied in the search for new plant derived natural products against these diseases and an overview on natural products of terpenoid origin with antiprotozoal potential were given. The present part II compiles the current knowledge on natural products with antiprotozoal activity that are derived from the shikimate pathway (lignans, coumarins, caffeic acid derivatives), quinones of various structural classes, compounds formed via the polyketide pathways (flavonoids and related compounds, chromenes and related benzopyrans and benzofurans, xanthones, acetogenins from Annonaceae and polyacetylenes) as well as the diverse classes of alkaloids. In total, both parts compile the literature on almost 900 different plant-derived natural products and their activity data, taken from over 800 references. These data, as the result of enormous efforts of numerous research groups world-wide, illustrate that plant secondary metabolites represent an immensely rich source of chemical diversity with an extremely high potential to yield a wealth of lead structures towards new therapies for NTDs. Only a small percentage, however, of the roughly 200,000 plant species on earth have been studied chemically and only a small percentage of these plants or their constituents has been investigated for antiprotozoal activity. The repository of plant-derived natural products hence deserves to be investigated even more intensely than it has been up to present.

The potential of secondary metabolites from plants as drugs or leads against protozoan neglected diseases - part I.

Infections with protozoan parasites are a major cause of disease and mortality in many tropical countries of the world. Diseases caused by species of the genera Trypanosoma (Human African Trypanosomiasis and Chagas Disease) and Leishmania (various forms of Leishmaniasis) are among the seventeen "Neglected Tropical Diseases" (NTDs) defined as such by WHO due to the neglect of financial investment into research and development of new drugs by a large part of pharmaceutical industry and neglect of public awareness in high income countries. Another major tropical protozoan disease is malaria (caused by various Plasmodium species), which -although not mentioned currently by the WHO as a neglected disease- still represents a major problem, especially to people living under poor circumstances in tropical countries. Malaria causes by far the highest number of deaths of all protozoan infections and is often (as in this review) included in the NTDs. The mentioned diseases threaten many millions of lives world-wide and they are mostly associated with poor socioeconomic and hygienic environment. Existing therapies suffer from various shortcomings, namely, a high degree of toxicity and unwanted effects, lack of availability and/or problematic application under the life conditions of affected populations. Development of new, safe and affordable drugs is therefore an urgent need. Nature has provided an innumerable number of drugs for the treatment of many serious diseases. Among the natural sources for new bioactive chemicals, plants are still predominant. Their secondary metabolism yields an immeasurable wealth of chemical structures which has been and will continue to be a source of new drugs, directly in their native form and after optimization by synthetic medicinal chemistry. The current review, published in two parts, attempts to give an overview on the potential of such plant-derived natural products as antiprotozoal leads and/or drugs in the fight against NTDs.

The potential of secondary metabolites from plants as drugs or leads against protozoan neglected diseases - part I

Infections with protozoan parasites are a major cause of disease and mortality in many tropical countries of the world. Diseases caused by species of the genera Trypanosoma (Human African Trypanosomiasis and Chagas Disease) and Leishmania (various forms of Leishmaniasis) are among the seventeen "Neglected Tropical Diseases" (NTDs) defined as such by WHO due to the neglect of financial investment into research and development of new drugs by a large part of pharmaceutical industry and neglect of public awareness in high income countries. Another major tropical protozoan disease is malaria (caused by various Plasmodium species), which -although not mentioned currently by the WHO as a neglected disease- still represents a major problem, especially to people living under poor circumstances in tropical countries. Malaria causes by far the highest number of deaths of all protozoan infections and is often (as in this review) included in the NTDs. The mentioned diseases threaten many millions of lives world-wide and they are mostly associated with poor socioeconomic and hygienic environment. Existing therapies suffer from various shortcomings, namely, a high degree of toxicity and unwanted effects, lack of availability and/or problematic application under the life conditions of affected populations. Development of new, safe and affordable drugs is therefore an urgent need. Nature has provided an innumerable number of drugs for the treatment of many serious diseases. Among the natural sources for new bioactive chemicals, plants are still predominant. Their secondary metabolism yields an immeasurable wealth of chemical structures which has been and will continue to be a source of new drugs, directly in their native form and after optimization by synthetic medicinal chemistry. The current review, published in two parts, attempts to give an overview on the potential of such plant-derived natural products as antiprotozoal leads and/or drugs in the fight against NTDs.

Reuse of sugarcane bagasse ash (SCBA) to produce ceramic materials.

Sugarcane bagasse ash (SCBA) is a residue resulting from the burning of bagasse in boilers in the sugarcane/alcohol industry. SCBA has a very high silica concentration and contains aluminum, iron, alkalis and alkaline earth oxides in smaller amounts. In this work, the properties of sintered ceramic bodies were evaluated based on the concentration of SCBA, which replaced non-plastic material. The ash was mixed (up to 60 wt%) with a clayed raw material that is used to produce roof tiles. Prismatic probes were pressed and sintered at different temperatures (up to 1200 °C). Technological tests of ceramic probes showed that the addition of ash has little influence on the ceramic properties up to 1000 °C. X-ray diffraction and thermal analysis data showed that, above this temperature the ash participates in the sintering process and in the formation of new important phases. The results reported show that the reuse of SCBA in the ceramic industry is feasible.

A proposal for chemical characterization and quality evaluation of botanical raw materials using glandular trichome microsampling of yacón (Polymnia sonchifolia, Asteraceae), an Andean medicinal plant

A proposal for chemical characterisation and quality evaluation of botanical raw materials by analysing the glandular trichomes from the leaves of two different populations of yacón (Polymnia sonchifolia Poeppig & Endlicher, Asteraceae) is described. This species is an Andean medicinal plant and the tea prepared with their leaves displays hypoglycemic property. The method was based on the glandular trichome microsampling using LC/DAD. Qualitative chromatographic fingerprints of the glands were obtained by isocratic runs and then compared between the two populations, showing the same qualitative profile. The two main metabolites detected in the glands were later isolated from a leaf rinse extract by preparative separation and identified as the melampolides enhydrin and uvedalin. These compounds, subtypes of sesquiterpene lactones, were used as chemical markers. This technique is applicable to other medicinal plants and can be used for the evaluation of the quality of dried material or leaf rinse extracts.

Neste trabalho é descrita uma proposta para a caracterização química e avaliação da qualidade de drogas vegetais através da análise de tricomas glandulares das folhas de duas populações diferentes de yacón (Polymnia sonchifolia Poeppig & Endlicher, Asteraceae). Esta espécie é uma planta medicinal andina e o chá preparado com suas folhas apresenta propriedade hipoglicemiante. O método baseou-se na microamostragem de tricomas glandulares, empregando-se cromatografia líquida de alta eficiência e detecção por arranjo de diodos. As impressões digitais dos cromatogramas das glândulas foram obtidas em sistema isocrático e comparadas entre si, resultado em um mesmo perfil qualitativo para ambas as populações avaliadas. Os dois metabólitos principais encontrados nas glândulas foram isolados através de métodos preparativos a partir de um extrato de lavagem foliar, sendo identificados como enidrina e uvedalina. Estas substâncias, subtipos de lactonas sesquiterpênicas, foram utilizadas como marcadores químicos. Esta técnica é aplicável a outras plantas medicinais e pode ser usada para avaliar a qualidade de materiais secos ou extratos foliares.

Immune and hormonal activity in adults suffering from depression.

An association between depression and altered immune and hormonal systems has been suggested by the results of many studies. In the present study we carried out immune and hormonal measurements in 40 non-medicated, ambulatory adult patients with depression determined by CID-10 criteria and compared with 34 healthy nondepressed subjects. The severity of the condition was determined with the Hamilton Depression Rating Scale. Of 40 depressed patients, 31 had very severe and 9 severe or moderate depression, 29 (72.5%) were females and 11 (27.5%) were males (2.6:1 ratio). The results revealed a significant reduction of albumin and elevation of alpha-1, alpha-2 and beta-globulins, and soluble IL-2 receptor in patients with depression compared to the values obtained for nondepressed subjects (P<0.05). The decrease lymphocyte proliferation in response to a mitogen was significantly lower in severely or moderately depressed patients when compared to control (P<0.05). These data confirm the immunological disturbance of acute phase proteins and cellular immune response in patients with depression. Other results may be explained by a variety of interacting factors such as number of patients, age, sex, and the nature, severity and/or duration of depression. Thus, the data obtained should be interpreted with caution and the precise clinical relevance of these findings requires further investigation.

Immune and hormonal activity in adults suffering from depression

An association between depression and altered immune and hormonal systems has been suggested by the results of many studies. In the present study we carried out immune and hormonal measurements in 40 non-medicated, ambulatory adult patients with depression determined by CID-10 criteria and compared with 34 healthy nondepressed subjects. The severity of the condition was determined with the Hamilton Depression Rating Scale. Of 40 depressed patients, 31 had very severe and 9 severe or moderate depression, 29 (72.5 percent) were females and 11 (27.5 percent) were males (2.6:1 ratio). The results revealed a significant reduction of albumin and elevation of alpha-1, alpha-2 and beta-globulins, and soluble IL-2 receptor in patients with depression compared to the values obtained for nondepressed subjects (P<0.05). The decrease lymphocyte proliferation in response to a mitogen was significantly lower in severely or moderately depressed patients when compared to control (P<0.05). These data confirm the immunological disturbance of acute phase proteins and cellular immune response in patients with depression. Other results may be explained by a variety of interacting factors such as number of patients, age, sex, and the nature, severity and/or duration of depression. Thus, the data obtained should be interpreted with caution and the precise clinical relevance of these findings requires further investigation

Sesquiterpene lactones and a myoinositol from glandular trichomes of Viguiera quinqueremis (Heliantheae; Asteraceae).

The extract of the floral parts of Viguiera quinqueremis afforded, in addition to known compounds, six new sesquiterpene lactones as well as a new myoinositol derivative. All compounds were detected in glandular trichomes which were collected micromechanically from the anther appendages and were analyzed by HPLC. Structure identification was performed by 1H NMR measurements including LC NMR and LC MS experiments.

Diterpenes and synthetic derivatives from Viguiera aspillioides with trypanomicidal activity.

The dichloromethane extract of the tuberous roots of Viguiera aspillioides was tested in vitro against T. cruzi and then investigated in order to identify its active compounds, which were the known diterpenes (-)-ent-kaur-16-en-19-oic acid, (-)-trachyloban-19-oic acid, and (-)-kauran-16alpha-ol. Synthetic derivatives of the acidic compounds were obtained and tested; one of them, (-)-kaur-16-en-19-ol, was also active. Their IC50 are given.

[Gastric neoplasm associated with IgA deficiency. Importance of multidisciplinary care]. / Neoplasia gástrica associada a défice de IgA. Importância da Multidisciplinaridade de Cuidados.

We present the case of 55-year old female patient with a rare malignant disease of the stomach associated with IgA deficiency. For its management the surgical, transfusional and renal support strategy required good coordination of pre, intra and post-operative care, and good articulation among the various sub-specialties. The patient experienced two severe anaphylactoid transfunctional reactions. Congenital IgA deficiency was detected with probable anti-IgA immunization associated with a metacrone adenocarcinoma of the stomach and nephrotic syndrome with renal failure. The transfusion methods utilized are discussed, and the results of the study aimed at detecting a compatible donor, with identical deficit, among family members are presented. In the absence of a national registry of donors with IgA deficiency, the prevention of these potentially dangerous reactions imposes a burden on the Service of Immuno-Hematherapy to take special measures on its own. This case points out the need to implement a blood bank with rare groups and IgA deficient blood, for homo/autologous transfusions.

Neonatal treatment with naloxone causes permanent hyperalgesia in rats.

The effect of treatment with naloxone early in life on pain responsiveness was studied in Wistar rats. Litters of six rats were divided equally into groups of 3 pups receiving daily naloxone (50 mg/kg, sc) and 3 pups receiving saline from the 3rd to 18th day of life. On days 30, 50, 70 and 90, one group of animals previously injected during suckling with naloxone (N = 21) and another with saline (N = 21) were submitted to the hot-plate test to measure the latency to paw licking. Other groups of rats also treated during suckling with naloxone (N = 13) and saline (N = 14) were assessed for the antinociceptive effect of morphine (10 mg/kg,sc). The naloxone group displayed a lower latency than the saline group in all test sessions and a diminished analgesic response to morphine. The results indicate that the use of naloxone (an antagonist opioid) during suckling, the brain growth spurt period, facilitates a long-lasting increased pain responsiveness and alters antialgesic mechanisms. In this respect, the opioid and non-opioid effects of naloxone on the ontogeny of neural systems should be taken into account.

Neonatal treatment with naloxone causes permanent hyperalgesia in rats

The effect of treatment with naloxone early in life on pain responsiveness was studied in Wistar rats. Litters of six rats were divided equally into groups of 3 pups receiving daily naloxone (50 mg/kg, sc) and 3 pups receiving saline from the 3rd to 18th day of life. On days 30, 50, 70 and 90, one group of animals previously injected during suckling with naloxone (N = 21) and another with saline (N = 21) were submitted to the hot-plate test to measure the latency to paw licking. Other groups of rats also treated during suckling with naloxone (N = 13) and saline (N = 14) were assessed for the antinociceptive effect of morphine (10 mg/kg,sc). The naloxone group displayed a lower latency than the saline group in all test sessions and a diminished analgesic response to morphine. The results indicate that the use of naloxone (an antagonist opioid) during suckling, the brain growth spurt period, facilitates a long-lasting increased pain responsiveness and alters antialgesic mechanisms. In this respect, the opioid and non-opioid effects of naloxone on the ontogeny of neural systems should be taken into account

Sciatic nerve conduction velocity of malnourished rats fed the human "basic regional diet" of the northeast of Brazil.

1. Rats were fed the "basic regional diet" (BRD) utilized by some human populations in northeastern Brazil, which contains only 7.8% protein (mainly from beans), to produce a form of malnutrition. The effects of chronic malnutrition on nerve impulse conduction velocity was studied in the sciatic nerve of young (121-143 days) and old (420-540 days) rats. 2. BRD rats presented a 50% reduction in sciatic nerve conduction velocity compared to controls fed a diet containing 22% casein. 3. Rats chronically fed a commercially available diet (CD group) containing an intermediate level of protein (14%) presented conduction velocities intermediate between values obtained from control and BRD rats (62% to 76% of the control values). 4. The conduction velocities of animals belonging to the same nutritional group were independent of sex, body weight or age. 5. Nutritional restriction imposed only in adult life by increasing the number of rats per cage in the casein group did not have any effect on the conduction velocity of nerve impulses. 6. The severity of the effects observed in the malnourished rats may be related to the quantity and quality of protein in the diet. In this respect, the present data agree with those reported in the literature on human and animal experiments. 7. The present results justify the use of the foods ingested by malnourished human populations for the development of experimental animal models of malnutrition.