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Biomedical subjects

L Peña

Publications and source records attributed to L Peña.

At least 19 recordsLinked to original sources

The effect of dexamethasone on disruption of ovarian steroid levels and receptors in female rats.

This study was conducted to investigate if the injection of a single dose of dexamethasone may cause disruption of adult female rat gonadal function in terms of plasma and ovarian level of both androgen and estrogen, ovarian morphology, and changes in localization of androgen, estrogen and glucocorticoid receptors. Adult female Long Evans rats (n=50, 250-300 g) were used. At day 0 rats received subcutaneously 1 ml of saline (n=25; control group) or dexamethasone at 0.1 mg/kg (n=25, treated group). Rats were sacrificed in groups of five on days 10, 15, 20, 25 and 30 after injection. Blood samples and one ovary were collected to analyze dexamethasone, 17beta-estradiol (E2), testosterone (T) and androstenedione (A4) concentrations by amplified EIA. The remaining ovary was removed and processed for histopathology and immunocytochemistry. Differences between individual means were analyzed by Pairwise t-test and Bonferroni post test to asses whether values presented statistical significance. Increased E2, T and A4 levels were observed both in plasma and ovary samples in treated group when comparing with control (p< 0.01) at all days post-injection even when dexamethasone was undetectable. Ovarian morphology of treated group showed features compatible with female infertility. Inmmunolocalization of androgen and estrogen receptors showed that both were negative in treated group while controls showed highest positivity (AR +++, ER ++). Glucocorticoid receptor showed higher positivity in dexamethasone treated rats (GR ++) than in controls (GR +). Obtained results showed clear evidence that a single dose of dexamethasone may disrupt gonadal function in rats, and that possibly leads to infertility.

Androstenedione↗

Role of steroid hormones and prolactin in canine mammary cancer.

In several animal studies, prolactin has been found to be essential for mammary epithelial development, and its administration has been consistently shown to increase the rate of mammary tumours. High levels of steroid hormones have also been suggested to enhance mammary cancer development. The present study investigates the levels of the following hormones in serum and in tissue homogenates in dogs bearing canine mammary tumours: prolactin (PRL), progesterone (P4), dehydroepiandrosterone (DHEA), androstenedione (A4), testosterone (T), 17beta-estradiol (17beta-E2) and estrone sulfate (S04E1). Eighty mammary tumours (40 dysplasias and benign and 40 malignant tumours) from 32 female dogs, and 10 normal mammary glands from eight female dogs without history of mammary tumours, were analysed. Prolactin and steroid hormones in serum and tissue homogenates, were analysed by enzyme immunoassays (EIA) techniques, previously validated for this animal species. Levels of prolactin in tissue homogenates were significantly different between malignant and benign mammary tumours (p<0.01). Serum prolactin concentrations were lower in the control group as compared with the group of dogs with benign tumours and in dogs with malignant tumours (p=0.01). Serum prolactin levels in dogs with benign lesions were not significantly different than those obtained from dogs with malignant tumours. Levels of steroid hormones were significantly higher in malignant tumours compared with the benign tumours and normal mammary glands (p<0.01) both in serum and homogenate determinations. Our results suggest that the canine neoplastic mammary gland could be a source of prolactin. Our hypothesis is that both prolactin and steroid hormones are involved in the growth of canine mammary cancer, and that they might have an autocrine/paracrine role in the maintenance of this disease.

Androgens↗

Cellular and subcellular changes in muscle, neuromuscular junctions and nerves caused by bee (Apis mellifera) venom.

Biopsy specimens of cervico-scutular muscles obtained from animals injected with bee crude venom were prepared for electron microscopy studies. At 6 h from Apis mellifera venom injection, in mice under transmission electron microscopy, the muscular fibres presented different atrophy levels with increment of the intermyofibrillar spaces. Tubules and sarcoplasmic reticulum elements were altered, in some places only tubular fragments and an increment of the intermyofibrillar spaces were noticed as well as loss of fibre regularity and prominent triads. In subsarcolemma region, areas lacking myofibrils and mitochondria damages were observed. Muscular segmental necrosis and atrophy areas were observed. Neuromuscular junctions were altered. The number of synaptic vesicles was very variable and synaptic clefts showed irregularities. A decrease in the number and arrangement of the synaptic clefts, as well as free polysomes, suggesting regeneration processes, were also observed. The myelinic nerves exhibited in the axon or in the wall vacuolisation areas. The presence of severe muscular lesions, the finding of venom activities in the presynaptic region and the detection of damages in the neuromuscular junctions at different chronological stages of our experiments indicate that the bee venom is highly toxic for neuromuscular structures.

Animals↗

Steroid hormone profile of canine inflammatory mammary carcinoma: a preliminary study.

Inflammatory mammary carcinoma (IMC) is the most aggressive spontaneous type of mammary malignant tumor both in women and dogs. Latest studies in dogs indicate that different endocrine mechanisms seem to be involved in inflammatory carcinomas (IMCs). The aim of the present study was to characterize the steroid hormone profile of inflammatory carcinoma, and to compare it with mammary dysplasias, benign tumors and other malignant tumors. Eighty-six mammary samples (10 normal mammary tissue, 21 dysplasias, 26 benign, 22 malignant, and 7 IMC) from 30 female dogs were used. Hormone levels of progesterone (P4), 17beta-estradiol (E2), androstenedione (A4), dehydroepiandrosterone (DHEA), and estrone sulphate (E1SO4) in tissue homogenates were measured by enzyme immunoassays (EIAs) techniques, previously validated for this species. IMC displayed the following steroid profile: P4: 13.80+/-0.56 microg/g; E2: 675.19+/-33.00 ng/g; A4: 631.73+/-70.73 microg/g; DHEA: 702.22+/-89.93 microg/g, and E1SO4: 2.84+/-0.32 mg/g. All of these hormones were significantly higher (P<0.001) compared with the hormone steroid profile determined for malignant, benign, dysplasias, and normal mammary tissue. The most relevant finding was the increased levels, two or three times, of both DHEA and E1SO4 in IMC respect to other groups (P<0.001). These results, together with the highest immunohistochemical expression of P450scc found in IMC, suggest the hypothesis that an autocrine mechanism could be especially involved in the development of canine inflammatory carcinoma.

Androstenedione↗

BRCA1 expression in canine mammary dysplasias and tumours: relationship with prognostic variables.

BRCA1 is a nuclear phosphoprotein that participates in the regulation of the cell cycle. The role of the BRCA1 gene in canine mammary tissue and mammary tumours has not been studied. The present study examined immunohistochemically the expression and intracellular distribution of BRCA1 protein in two normal, seven dysplastic and 44 neoplastic canine mammary glands and its relationship with clinical and pathological variables and other prognostic parameters. Strong nuclear immunolabelling of BRCA1 protein was observed in the epithelial cells of the normal mammary glands and mammary dysplasias. The majority of benign tumours, and more especially of malignant tumours, showed a significant reduction in the nuclear expression of BRCA1 protein and an increase in cytoplasmic expression. Loss of BRCA1 expression was associated with high proliferation marker Ki-67 and ER-alpha negative tumours. The reduction and aberrant distribution of BRCA1 in canine mammary tumours were significantly associated with malignant characteristics. The results may indicate that BRCA1 has a role in the malignant behaviour of these tumours.

Adenocarcinoma↗

Regeneration of transgenic citrus plants under non selective conditions results in high-frequency recovery of plants with silenced transgenes.

Insertion of foreign DNA into plant genomes frequently results in the recovery of transgenic plants with silenced transgenes. To investigate to what extent regeneration under selective conditions limits the recovery of transgenic plants showing gene silencing in woody species, Mexican lime [ Citrus aurantifolia (Christm.) Swing.] plants were transformed with the p25 coat protein gene of Citrus tristeza virus (CTV) with or without selection for nptII and uidA. Strikingly, more than 30% of the transgenic limes regenerated under non-selective conditions had silenced transgenes, and in all cases silencing affected all the three transgenes incorporated. These results indicate that the frequency of transgene silencing may be greatly underestimated when the rate of silencing is estimated from the number of regenerants obtained under selective conditions. To our knowledge, this is the first report in which the frequency of gene silencing after transformation has been quantified. When the integration pattern of T-DNA was analyzed in silenced and non-silenced lines, it was observed that inverted repeats as well as direct repeats and even single integrations were able to trigger gene silencing. Gene silencing has often been associated with the insertion of DNA sequences as inverted repeats. Interestingly, here, direct repeats and single-copy insertions were found in both silenced and non-silenced lines, suggesting that the presence of inverted-repeat T-DNAs and the subsequent formation of dsRNAs triggering gene silencing cannot account for all silencing events.

Citrus↗

Green fluorescent protein as a visual marker in somatic hybridization.

Using a transgenic citrus plant expressing Green Fluorescent Protein (GFP) as a parent in somatic fusion experiments, we investigated the suitability of GFP as an in vivo marker to follow the processes of protoplast fusion, regeneration and selection of hybrid plants. A high level of GFP expression was detected in transgenic citrus protoplasts, hybrid callus, embryos and plants. It is demonstrated that GFP can be used for the continuous monitoring of the fusion process, localization of hybrid colonies and callus, and selection of somatic hybrid embryos and plants.

Biomarkers↗

Use of aminoglutethimide in the treatment of pituitary-dependent hyperadrenocorticism in the dog.

The aim of this study was to evaluate the efficacy and safety of aminoglutethimide in the treatment of dogs with pituitary-dependent hyperadrenocorticism (PDH). Ten dogs were diagnosed with PDH based on clinical and laboratory data, adrenal function tests (adrenocorticotropic hormone [ACTH] stimulation test and urinary cortisol/creatinine ratio [UCCR] combined with a high dose oral dexamethasone suppression test) and ultrasonographic evaluation of the adrenal glands. Aminoglutethimide was administered daily at a dose of 15 mg/kg bodyweight for one month. Median basal cortisol concentration and post-ACTH cortisol concentration one month after treatment were significantly lower than pretreatment values. Complete response was achieved in one dog, and partial response was obtained in three dogs. Severe side effects of anorexia, vomiting and weakness occurred in one dog and medication was withdrawn. Two further dogs developed decompensations of concurrent diseases and medication was stopped in these animals as well. Mild toxicity occurred in four dogs. Moderate to severe elevations in liver enzymes occurred in all dogs. The efficacy of this drug is lower than that observed using mitotane and ketoconazole, and adverse effects limit its use. Aminoglutethimide, using the protocol described, cannot be recommended for long-term management of PDH in the dog.

Adrenal Glands↗

Lingual calcinosis circumscripta in a dog.

Calcinosis circumscripta occurs rarely in dogs and cats, especially in the oral cavity. This case report describes the diagnosis and management of lingual calcinosis circumscripta in an eight-month-old, intact/male German shepherd-cross dog. An excellent prognosis for this disease may be achieved when the patient is managed as described in this report.

Animals↗

Inflammatory mammary carcinoma in dogs: 33 cases (1995-1999).

OBJECTIVE: To determine epidemiologic, clinical, and pathologic characteristics of inflammatory mammary carcinoma (IC) in dogs. DESIGN: Retrospective study. ANIMALS: 33 dogs with IC and 153 dogs with malignant mammary tumors other than IC. PROCEDURES: Medical records were reviewed, and signalment, history, physical examination findings, and results of thoracic radiography and necropsy were obtained. RESULTS: 33 of 436 (7.6%) dogs examined at a veterinary teaching hospital because of dysplasia or tumors of the mammary glands and 33 of 186 (17.7%) dogs with at least 1 malignant tumor had IC. Thirty-two of the 33 dogs were sexually intact. Dogs with IC were significantly older than were dogs with other malignant mammary tumors, and in dogs with IC, the tumor was initially noticed a mean of 52 days after the beginning of the last observed estrus, whereas in dogs with other mammary tumors, the tumor was initially noticed a mean of 137 days after the beginning of the last observed estrus. Dogs with IC were more likely to be anorectic and to have generalized weakness, weight loss, and thoracic metastases. Dogs with IC survived a mean of 25 days with palliative treatment. Histologically, involvement of dermal lymphatic vessels was identified in 14 of 19 (74%) dogs with IC. Two clinical forms of IC (primary and secondary) were identified. Dogs with primary IC had a worse clinical condition. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that IC is an uncommon but distinct entity in dogs. A histologic finding of dermal lymphatic involvement should be considered a hallmark for the pathologic diagnosis of IC in dogs.

Age Factors↗

Recent advances in the genetic transformation of trees.

As the commercial production of transgenic annual crops becomes a reality in many parts of the world, many people wonder if the genetic engineering of perennial trees will allow their eventual commercialization. Not long ago, trees were considered to be recalcitrant material for most molecular biology techniques, including genetic transformation. However, transgenes for shortening the juvenile phase or for phytoremediation purposes have now been incorporated, and the alteration of lignin biosynthesis and increased cellulose accumulation in forest trees have also been accomplished. For long-lived tree species, new questions arise regarding the stability of integration and expression of foreign genes. Biosafety considerations, including transgene dispersion through the pollen and advances in strategies to avoid this, are also important.

Biodegradation, Environmental↗

Constitutive expression of Arabidopsis LEAFY or APETALA1 genes in citrus reduces their generation time.

Citrus trees have a long juvenile phase that delays their reproductive development by between 6 and 20 years, depending on the species. With the aim of accelerating their flowering time, we transformed juvenile citrus seedlings to constitutively express the Arabidopsis LEAFY (LFY) or APETALA1 (AP1) genes, which promote flower initiation in Arabidopsis. Both types of transgenic citrus produced fertile flowers and fruits as early as the first year, notably through a mechanism involving an appreciable shortening of their juvenile phase. Furthermore, expression of AP1 was as efficient as LFY in the initiation of flowers, and did not produce any severe developmental abnormality. Both types of transgenic trees flowered in consecutive years, and their flowering response was under environmental control. In addition, zygotic and nucellar derived transgenic seedlings had a very short juvenile phase and flowered in their first spring, demonstrating the stability and inheritance of this trait. These results open new possibilities for domestication, genetic improvement, and experimental research in citrus and other woody species.

Arabidopsis↗

Factors influencing the incidence and prognosis of canine mammary tumours.

Factors relating to the incidence of canine mammary tumours are reviewed. Increased age, intact status or ovariectomy after 2.5 years of age, as well as progestagen treatment, can all lead to an increased risk of mammary neoplasia in the bitch. In addition, obesity early in life, and a habitual diet based on home-made food (rich in beef and pork, and poor in chicken) as opposed to commercial food, are also associated with the occurrence of mammary tumours. Other aspects related to incidence are also discussed. Increased age at diagnosis, invasive growth (fixed to adjacent tissues), large tumour size, ulceration of skin, and axillary or inguinal node involvement are clinical parameters associated with a low chance of survival after surgical excision of mammary tumours. Histological typing and grading of the tumour allows the establishment of a prognosis, which is poor where there is tumour proliferation as measured by S-phase fraction determination and Ki-67 immunostaining.

Age Factors↗

Immunohistologic detection of estrogen receptor alpha in canine mammary tumors: clinical and pathologic associations and prognostic significance.

Eighty-nine canine mammary tumors and dysplasias of 66 bitches were investigated to determine the immunohistochemical expression of classical estrogen receptor (ER-alpha) and its clinical and pathologic associations and prognostic value. A complete clinical examination was performed and reproductive history was evaluated. After surgery, all animals were followed-up for 18 months, with clinical examinations every 3-4 months. ER-alpha expression was higher in tumors of genitally intact and young bitches (P < 0.01, P < 0.01) and in animals with regular estrous periods (P = 0.03). Malignant tumors of the bitches with a previous clinical history of pseudopregnancy expressed significantly more ER-alpha (P = 0.04). Immunoexpression of ER-alpha decreased significantly with tumor size (P = 0.05) and skin ulceration (P = 0.01). Low levels of ER-alpha were significantly associated with lymph node involvement (P < 0.01). Malignant tumors had lower ER-alpha expression than did benign tumors (P < 0.01). Proliferation index measured by proliferating cell nuclear antigen immunostaining was inversely correlated with ER-alpha scores (P = 0.05) in all tumors. Low ER-alpha levels in primary malignant tumors were significantly associated with the occurrence of metastases in the follow-up (P = 0.03). Multivariate analyses were performed to determine the prognostic significance of some follow-up variables. ER-alpha value, Ki-67 index, and age were independent factors that could predict disease-free survival. Lymph node status, age, and ER-alpha index were independent prognostic factors for the overall survival. The immunohistochemical detection of ER-alpha in canine mammary tumors is a simple technique with prognostic value that could be useful in selecting appropriate hormonal therapy.

Animals↗

Pathological alterations caused by Anoplocephala perfoliata infection in the ileocaecal junction of equids.

The pathological alterations caused by Anoplocephala perfoliata in the ileocaecal junction of 28 equids slaughtered in an abattoir in Madrid (Central Spain) are described. The lesions were scored in grades based on the intensity of the damage and were related to the tapeworm number observed. The first grade (grade I) of alterations consisted of a slight enteritis associated with focal erosions observed in 43% of parasitized animals with low parasitic burden (1-26 tapeworms). The second grade (grade II) was a focal pseudomembranous enteritis, present in the ileocaecal junctions of 36% infected animals with moderate to high burden (23-188 tapeworms), and the third grade (grade III) was a regional necrotizing enteritis, present in the animals (21%) with the highest burden (72-248 tapeworms). The possible role of the lesions caused by this parasite in the aetiology of colic is discussed.

Animals↗

An unusual late-onset case of propionic acidaemia: biochemical investigations, neuroradiological findings and mutation analysis.

UNLABELLED: We report a 5-year-old boy with propionic acidaemia who developed a rapidly fatal necrosis of the basal ganglia after an episode of clinical deterioration. Neither metabolic acidosis nor hyperammonaemia were present. Organic acid analysis in both urine and CSF showed increased levels of methylcitric and 3-hydroxypropionic acids. Propionic acidaemia was confirmed by demonstrating a propionyl-CoA carboxylase deficiency (11% of control value) in skin fibroblasts. DNA analysis revealed that the patient was a compound heterozygote for two mutations in the PCCB gene. CONCLUSION: Propionic acidaemia can present as a sudden and fatal neurological disease and not only as an organic aciduria with severe biochemical dis-turbances and progressive neurological deterioration.

Amino Acid Metabolism, Inborn Errors↗