Immunolocalization of estrogen and progesterone receptors in neuroendocrine tumors of lung, skin, gastrointestinal and female genital tracts

Expression of estrogen (ER) and progesterone (PR) receptors has traditionally been associated with hormone-responsive organs, such as breast, ovary, and endometrium, and carcinomas arising therefrom. More recently, examples of ''unexpected'' ER or PR expression have been reported, particularly in tumors of endocrine tissues, such as thyroid and pancreatic islet cells. We tested the hypothesis that neuroendocrine tumors of various primary and metastatic sites might also express ER or PR or both by performing a retrospective immunohistochemical study in a series of 59 formalin- or mechacarn-fixed neuroendocrine carcinomas of various sites, including lung, skin, gastrointestinal and female genital tracts, and including carcinoid and atypical carcinoid tumors, small cell carcinomas, and Merkel cell carcinomas. We employed the anti-ER monoclonal antibody 1D5 and the anti-PR monoclonal antibody PgR1A6 using standard immunohistochemical techniques after microwave-based heat-induced epitope retrieval. Two of 28 carcinoid tumors demonstrated ER positivity; six of 30 cases were positive for progesterone receptor only. In addition, PR expression was found in one of two cases of atypical carcinoid, in five of 25 cases of small cell carcinoma, and in one of two cases of Merkel cell carcinoma. None of the atypical carcinoids, small cell carcinomas, or Merkel cell carcinomas were ER positive. In most cases, the fraction of tumor cell nuclei that were positive was <50%. These studies add the spectrum of neuroendocrine tumors that can express these hormone receptors. Similar to the pattern previously described in the subsets of meningiomas and islet cell tumors, PR but not ER is detectable in most cases. These results underscore the caution that should be exercised in determining tissue origin of metastatic carcinomas based only on detection of hormone receptors by immunohistochemistry.

AGE-RELATED-CHANGES OF GLUTATHIONE CONTENT, GLUTATHIONE-REDUCTASE AND GLUTATHIONE-PEROXIDASE ACTIVITY OF HUMAN ERYTHROCYTES

1. In order to investigate the effect of aging on the erythrocyte glutathione system, total glutathione (GSH), glutathione reductase (GSH-red) and glutathione peroxidase (GSH-px) levels were measured in erythrocytes from 33 young (mean age = 30.5 +/- 9.7 years) and 28 aged (mean age = 68.9 +/- 11.4 years) healthy individuals.2. GSH was 3.5 +/- 1.8-mu-M/g Hb for the young group, a value significantly greater (P < 0.01) than 2.3 +/- 0.9-mu-M/g Hb found for the aged group. Similarly, GSH-red activity, 5.5 +/- 1.8 IU/g Hb, was higher (P < 0.05) for the young group than 3.4 +/- 0.9 IU/g Hb found for the aged group. The GSH-px activity levels for the young group, 21.1 +/- 5.9 IU/g Hb, were significantly greater (P < 0.01) than 12.0 +/- 3.3 IU/g Hb for the aged group. The lower activity detected in the aged group for all of these parameters of the glutathione redox system was not related to low levels of hematocrit or hemoglobin.3. There was no statistical difference in the activation coefficient (AC) of reductase (+FAD/-FAD) between groups, which seems to indicate that the lower activity of glutathione reductase observed in the aged group was not due to riboflavin deficiency.4. Additional information is required to determine the mechanisms controlling the glutathione redox system and its role in the aging process.

Effects of Short-term Hyper- and Hypoprolactinaemia on Hormones of the Pituitary, Gonad and -Thyroid Axis and on Semen Quality in Male Beagles

Effects of a short-term hyper- and hypoprolactinaemia on serum concentrations of LH, testosterone and semen quality in six male Beagles were investigated. Blood samples were collected at 3-day intervals for 12 weeks. The time span was divided into five 3-week periods: pre-treatment, metoclopramide (MCP) treatment (0.2 mg/kg orally three times daily), cabergoline (CAB) treatment (5 mu g/kg orally once daily), post-treatment 1 and post-treatment 2. In the latter, only semen characteristics were evaluated. Semen parameters were analyzed once per week during the whole 15-week investigation time. At the end of each period, the effects of a single intravenous injection of thyrotropin-releasing hormone (TRH; 10 mu g/kg) on the secretion of prolactin (PRL), LH, testosterone, thyroid-stimulating hormone and thyroxine (T4) were investigated. Pre-treatment serum PRL concentration increased under MCP (p < 0.05), followed by a decrease under CAB administration (p < 0.05). Luteinizing hormone and testosterone concentrations were not affected. Except for straight-line sperm velocity, semen quality did not differ between collection periods. A single iv TRH injection induced a significant PRL increase at 20 min in all experimental periods except during CAB treatment. Luteinizing hormone and testosterone did not show clear TRH-related changes. Basic T4 levels were significantly reduced after CAB treatment ( p < 0.05). The results of the present study demonstrate that MCP-induced short-term hyperprolactinaemia in male beagles does not seriously affect the hypothalamo-pituitary axis and semen quality.

Comparison of two immunochromatographic assays and the indirect immunofluorscence antibody test for diagnosis of Trypanosoma cruzi infection in dogs in south central Louisiana

Two rapid tests evaluated in dogs considered to be of high risk of Infection with the Chagas parasite Trypanosoma cruzi using two immunochromatographic assays. Trypanosoma Detect (TM) for canine, InBios, Seattle, WA and CHAGAS STAT-PAK (TM) assay, Chembio Diagnostic Systems, Medford, NY, in south central Louisiana. For this purpose a serological survey was carried out in a total of 122 dogs and a serum bank was created. These 122 animals were first tested by IFAT that was used as the standard test From the serum bank 50 samples were tested using the two rapid Chagas assays and results compared to the standard test IFAT The serological survey using IFAT showed it prevalence of T cruzi infection in 22.1% of the tested dogs. In the immunochromatographic assays. 13 and 11 animals were positive on rapid assay Trypanosoma Detect (TM) for canine, InBios and CHAGAS STAT-PAK (TM), Chembio Diagnostic Systems, respectively compared to 11 positive by IFAT. These two immunochromatographic tests have shown high susceptibility and specificity compared to our standard method IFAT. The rapid, easy and accurate screening assays used in conjunction with confirmatory tests, would be an excellent tool for veterinarians to diagnose T cruzi infection. Early detection of T cruzi infection may prevent complications through an effective treatment. Greater awareness by veterinarians of the risk. clinical findings, history along with diagnostic methods will contribute greatly to an understanding of the true prevalence of Chagas disease in dogs in Louisiana. (c) 2009 Elsevier B.V. All rights reserved.

Effect of thermal stress on physiological parameters, feed intake and plasma thyroid hormones concentration in Alentejana, Mertolenga, Frisian and Limousine cattle breeds

The aim of the present study was to assess the heat tolerance of animals of two Portuguese (Alentejana and Mertolenga) and two exotic (Frisian and Limousine) cattle breeds, through the monitoring of physiological acclimatization reactions in different thermal situations characterized by alternate periods of thermoneutrality and heat stress simulated in climatic chambers. In the experiment, six heifers of the Alentejana, Frisian and Mertolenga breeds and four heifers of the Limousine breed were used. The increase in chamber temperatures had different consequences on the animals of each breed. When submitted to heat stress, the Frisian animals developed high thermal polypnea (more than 105 breath movements per minute), which did not prevent an increase in the rectal temperature (from 38.7 degrees C to 40.0 degrees C). However, only a slight depression in food intake and in blood thyroid hormone concentrations was observed under thermal stressful conditions. Under the thermal stressful conditions, Limousine animals decreased food intake by 11.4% and blood triiodothyronine (T3) hormone concentration decreased to 76% of the level observed in thermoneutral conditions. Alentejana animals had similar reactions. The Mertolenga cattle exhibited the highest capacity for maintaining homeothermy: under heat stressful conditions, the mean thermal polypnea increased twofold, but mean rectal temperature did not increase. Mean food intake decreased by only 2% and mean T3 blood concentration was lowered to 85,6% of the concentration observed under thermoneutral conditions. These results lead to the conclusion that the Frisian animals had more difficulty in tolerating high temperatures, the Limousine and Alentejana ones had an intermediate difficulty, and the Mertolenga animals were by far the most heat tolerant.

Evaluation of macrophage activity and antibody production in genetically selected mice infected with Leptospira serovar pomona

The aim of the present study was to evaluate the role of macrophage activity and antibody production in experimental infection with Leptospira Pomona in mice genetically selected for high (H) or low (L) humoral immune response. To evaluate macrophage activity, reactive oxygen and nitrogen intermediates were determined. Also, the production of tumor necrosis factor (TNF-alpha) and the recovery of Leptospira-specific antibodies in the kidneys and liver were assessed; histological lesions were analyzed using the hematoxylin-eosin technique, and Leptospira antigens in tissues were determined by immunohistochemistry. Results showed that recovery of microorganisms from the analyzed organs was lower in LIV-A mice. However, HIV-A animals showed total restraint since the 14th day after infection, whereas LIV-A mice still had bacteria in the liver at the 21st post-infection day. Immune response against Pomona serovar in those lineages was characterized as high production of antibodies, mainly in late periods of the infectious process. The production of reactive oxygen and nitrogen intermediates also contributed to the elimination of Leptospira Pomona in all two lineages; H2O2 production was an important factor in HIV-A mice, as well as NO production in the LIV-A animals, mainly at the latest post-inoculation periods. The same occurred regarding TNF-alpha production. Severe renal lesions were observed at periods in which larger numbers of leptospires were isolated using the culture technique. Tissue alterations persisted in LIV-A mice, even at periods in which leptospires were not recovered. Immunohistochemistry showed to be more sensitive than culturing. However, both techniques were appropriate for the agent identification in the studied lineages. Results suggest that such lineages could represent an important model to investigate pathogenesis and immune response against the varied serovars of leptospires.

Detection of cross infections by Leishmania spp. and Trypanosoma spp. in dogs using indirect immunoenzyme assay, indirect fluorescent antibody test and polymerase chain reaction

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of gonadectomy on prolactin and LH secretion and the pituitary-thyroid axis in male dogs

The effects of gonadectomy on the secretion of prolactin, LH, TSH, and thyroxine were investigated. Blood serum hormone concentrations were analysed before and at 20, 120, and 180 min after a single iv TRH injection in each of eight healthy intact and castrated male beagle dogs before (control) and after 4-week treatment with the dopamine-2 receptor agonist cabergoline. Under control conditions the mean prolactin, TSH, and thyroxine concentrations were similar in intact and gonadectomised dogs, and administration of TRH provoked a significant (p < 0.01) increase in concentrations of the three hormones. The overall inhibitory effect of cabergoline treatment on prolactin secretion was more pronounced in the castrated dogs compared with the intact group. Cabergoline significantly suppressed the TRH-induced prolactin increase in each group (p < 0.01). Corresponding TRH-stimulated TSH concentrations were not affected by cabergoline. In the gonadectomised dogs, thyroxine concentrations before and at 120 and 180 min after TRH injection were significantly lower than under control conditions. LH concentrations were always higher (p < 0.01) in gonadectomised dogs compared with the intact dogs, but appeared to be affected neither by TRH nor by cabergoline administration. It can thus be concluded from the results, that gonadectomy does not result in hyperprolactinaemia in male dogs, while LH concentrations are significantly increased due to missing androgen feedback. Thyroid function remains unaffected by gonadectomy. Testicular steroids appear to interact with central dopaminergic and probably other neuroendocrine mechanisms regulating the secretion of prolactin, TSH, and thyroxine. Thus, long-term dopamine-2 receptor agonistic treatment may lead to a hypothyroid condition in castrated male dogs. (c) 2009 Elsevier B.V. All rights reserved.

Maternal antibody transfer to broiler progeny varies among strains and is affected by grain source and cage density

Two experiments were conducted to examine the effects of broiler breeder dietary grain source and cage density on maternal antibody (MatAb) transfer to progeny in 2 genetic strains (A and B). Broiler breeders were assigned to 16 litter floor pens and fed either corn- or wheat-based diets. Breeders were administered 4 live vaccines against Newcastle disease virus (NDV). At 23 wk of age, pullets and cocks, which reflected the full BW distribution from each treatment, were moved to a cage breeder house and placed at 1 or 2 hens/cage. Breeders were artificially inseminated at 44 wk (experiment 1) and 52 wk of age (experiment 2). Eggs were collected for 8 d, incubated, and placed in individual pedigree bags at d 19 of incubation. Blood samples from 5 chicks per treatment combination were collected at hatch in both experiments. Spleen and bursa were collected from the same chicks for histomorphometry analyses in experiment 2. In the second experiment, 12 chicks per treatment were placed in cages. Progeny were provided diets based on the same grain (corn or wheat) as their parents. Serum samples were collected at 5, 9, and 13 d of age and analyzed for anti-NDV MatAb. Data were analyzed as a 2 x 2 x 2 factorial design considering strain, dietary grain source, and cage density as main factors. Interaction effects were observed in breeders and progeny. Experiment 1 showed that strain A chicks had lower levels of MatAb when hens were housed at 2 hens/cage rather than 1 hen/cage. The MatAb levels of strain B chickens were not affected by cage density in either experiment. Experiment 2 demonstrated similar effects of cage density on MatAb levels and the area of bursa follicles for both strains. Progeny of breeders fed corn-based diets had smaller spleen white pulp only when hens were housed at 2 hens/cage compared with 1 hen/cage. The results of these experiments suggest that breeder strain and cage-density conditions affected MatAb transfer to progeny and embryo development of spleen and bursa.

Magneto Immunoassays for Plasmodium falciparum Histidine-Rich Protein 2 Related to Malaria based on Magnetic Nanoparticles

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Real-time monitoring and kinetic parameter estimation of the affinity interaction of jArtinM and rArtinM with peroxidase glycoprotein by the electrogravimetric technique

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Follicular thyroid carcinoma in dogs. Report of cases

The treatment of choice for these cases was surgical removal. In both cases the diagnostic was infiltrative follicular carcinoma of thyroid. The first animal did not present a good recovery after the surgery, and died four hours after the procedure. In the second case, the diagnosis was more precocious and antineoplastic chemotherapy was used after the surgery. At the time of submission of this manuscript, this animal had survived for foully months. Currently, there is a need to define the protocols of chemotherapy to avoid relapse and metastases, in order to increase the life expectancy in dogs with thyroid neoplasm.