Gene Expression of Fresh and Frozen/Thawed Canine Embryos.

BACKGROUND: Canine embryo cryopreservation and subsequent transfer are relevant in the use of reproductive technologies. OBJECTIVE: The purpose of this study is the identification and quantification of the gene expression BAX and Bcl2, AQP3, Na+/K+ ATPase alpha-1 and beta-1 and LIFr in canine embryos obtained in vivo and after freezing. MATERIALS AND METHODS: For the collection of embryos, the bitches were identified at pro-estrous until the detection of 80-90% superficial cells. After that, they were artificially inseminated with fresh semen. The embryos were collected after ovariohysterectomy. RNA was extracted and amplified, and embryos were randomly distributed into fresh (Fr) and frozen/thawed (Ft) groups. RESULTS: Eighteen blastocysts were collected from three bitches. Genes BAX, AQP3 and LIFr did not differ among the studied groups. CONCLUSION: We suggest, through these results, that the genes BAX, Bcl2, AQP3, Na + / K + ATPase alpha-1 and beta-1 and LIFr were expressed in canine blastocysts collected in vivo and after slow freezing cryopreservation.

Expressão gênica dos REα, REβ e PR em tumores mamários de cadelas por meio do q-PCR / REα, REβ and PR gene expression in mammary gland tumors of female dogs by q-PCR

A expressão de receptores de estrógeno (ER) e progesterona (PR) por meio da técnica de q-PCR foi avaliada em 26 cadelas portadoras de neoplasias mamárias e cinco cadelas sem afecções mamárias (grupo controle). Os resultados mostraram que os três grupos de animais estudados - com tumor maligno ou benigno e controle - expressaram receptores de estrógeno alfa, beta e progesterona. A quantificação relativa mostrou tendência para uma expressão maior de receptores no grupo controle e menor no grupo de animais com neoplasias malignas. Além disso, observou-se expressão maior de ERα em relação ao ERβ, e as neoplasias malignas de origem mista apresentaram maiores concentrações dos receptores PR, ERα e ERβ que as neoplasias de origem epitelial.

The estrogen and progesterone receptor (ER and PR) expression with the q-PCR technique was evaluated in 26 female dog carrying of mammary tumors and five female dogs without mammary disease (control group). The results showed that the three animal groups evaluated - malignant or benign tumor and control - expressed alpha and beta estrogen and progesterone receptors. The relative quantification showed a tendency for a higher expression of receptors from the control group and smaller in the malignat tumors animal group. Also, there was a major ERα expression regarding to ERβ and the malignat tumors from mixed origin presented higher concentrations of receptors PR, ERα and ERβ, when compared to tumors of epithelial origin.

Imunoistoquímica de útero e cérvice de cadelas com diagnóstico de piometra / Immunohistochemical of the cervix and uterus of bitches diagnosed with pyometra

Analisou-se a piometra de 31 cadelas, de raças e idades variadas, sendo 25 cadelas com piometra de cérvice aberta e seis de cérvice fechada. Após ovariossalpingo-histerectomia, foram coletados fragmentos da cérvice e do útero para a avaliação imunoistoquímica. Foram analisados os receptores de estrógenos α e β, progesterona e colágenos I e III. Foram realizadas imunomarcações em diferentes regiões da cérvice, como o epitélio glandular, o epitélio luminal e o estroma glandular, assim como em diferentes regiões do útero, como o epitélio glandular e o estroma glandular. As imunomarcações de colágenos I e III foram realizadas nas regiões glandular e muscular da cérvice e do útero. Concentrações de receptores de progesterona foram maiores em cadelas com piometra fechada.

The pyometra in 31 bitches of different breeds and ages, 25 with pyometra cervix open and 6 closed cervix was analyzed. After the ovariohysterectomy procedure, samples were collected from the cervix and uterus to evaluate immunohistochemistry. For immunohistochemical evaluation estrogen receptors α and β, progesterone and collagen I and III were analyzed. Immunostainings were performed in different regions of the cervix such as glandular epithelium, luminal epithelium and glandular stroma, as well as in different regions of the uterus, such as glandular epithelium and glandular stroma. The immunostainings for collagen I and III were performed in muscular and glandular regions of the cervix and uterus. The concentration the progesterone receptors were elevated in bitches from the closed pyometra.

Ovarian activity reversibility after the use of deslorelin acetate as a short-term contraceptive in domestic queens.

The objective was to evaluate ovarian activity reversibility in domestic queens after short-term contraceptive treatment with deslorelin acetate. Ten mature queens were used. In all queens, the estrous cycle was evaluated every 72 h by vaginal cytology (VC) and behavior assessments. When queens had VC characteristic of interestrus or diestrus, one deslorelin acetate implant (4.7 mg) was placed in the subcutaneous tissue of the interscapular region (day of insertion = Day 0). Thereafter, VC was performed every 48 h and on Day 90, implants were removed. At Day 100, estrus and ovulation were induced with 100 IU eCG (im), followed by 100 IU hCG (im), 84 h later (Day 103.5). Queens were ovariohysterectomized on Day 106. Corpora lutea (CL) were counted, oviducts were flushed, and oocytes were identified, isolated and stained to assess viability. In all queens, blood samples for plasma progesterone concentrations were collected once a week, from Days -21 to 106. After deslorelin acetate application, four queens had VC and behavior typical of estrus, and one ovulated. Furthermore, ovulation occurred in three queens that did not have VC or behavior consistent with estrus. After the initial ovarian stimulation, all females had anestrous VC during the deslorelin treatment period. Implants were readily removed. Following implant removal, all females responded to treatments to induce estrus and ovulation. There were (mean ± SEM) 13.1 ± 5.5 CL and 8.1 ± 5.5 oocytes per queen; the oocyte recovery rate was 56.8 ± 25.4% and all recovered oocytes were viable. We concluded that deslorelin acetate can be used as a reversible short-term contraceptive in domestic cats, because estrus and ovulation were successfully induced following implant removal.

Progesterone (PR), oestrogen (ER-α and ER-ß) and oxytocin (OTR) gene expression in the oviduct and uterus of pregnant and non-pregnant bitches.

The aim was to assess hormone receptor gene expression in the oviduct and uterus during canine pregnancy. Nineteen pregnant bitches divided into four groups were ovariohysterectomized (OVH) at either day 8, 12, 21 or 60 of pregnancy, and five non-pregnant females underwent OVH 12 days after the pre-ovulatory Luteinizant Hormone (LH) surge and served as controls. RT-qPCR for progesterone (PR), oestrogen (ER-α and ER-ß) and oxytocin (OTR) receptors was performed on the oviduct and uterine tissue. The mRNA PR expression in the uterus during early stages of pregnancy and the luteal phase was higher than at other times. The mRNA ER-ß expression in the oviduct during early pregnancy was less than in non-pregnant bitches. In the uterus, the mRNA ER-ß expression was higher in the initial stages of pregnancy. The ER-α expression was higher in the oviduct and uterus in advanced stages of pregnancy. The mRNA OTR expression in the oviduct was lower than in the uterus in control group. The expression of this receptor in oviduct and the uterus was higher in the final stages of pregnancy, when compared with other phases. These data suggested that the serum progesterone concentrations probably exert a direct control on the PR and ER (α and ß) expression and indirectly on OTR expression in the bitch oviduct and uterus.

Lactate in bitches with pyometra.

Lactate is a compound produced by the anaerobic metabolism of glucose, and hyperlactataemia occurs when the rate of production of lactate exceeds the rate of elimination. This occurs in situations of hypoxia and tissue hypoperfusion. Lactate has been considered a useful prognostic indicator in critically ill patients. Pyometra is a disease of adult female dogs characterized by inflammation of the uterus with an accumulation of exudate, which occurs during the luteal phase. It is one of the most common diseases that occur in the genital tract of female dogs. A total of 31 dogs were diagnosed with pyometra. The diagnosis was confirmed at ultrasonography. Of the 31 dogs, 25 females had open cervix pyometra and six had closed cervix pyometra. Plasma lactate concentrations were determined by an enzymatic colorimetric method. The average concentration (±SD) of plasma lactate in all 31 bitches with pyometra was 3.55 ± 0.46 mm. Healthy dogs had plasma lactate concentrations between 0.3 and 2.5 mm (mean ± SD). Concentrations ranged from 0.8 to 2.9 mm when plasma lactate was measured with a portable device and 0.4-2.6 mm with the blood gas analyser. Even though plasma lactate values vary between several studies and equipment used to measure concentrations, our results for dogs with pyometra are higher indicating hyperlactataemia (Thorneloe et al. , Can Vet J 48, 283-288). Plasma lactate in dogs with closed cervix pyometra was mean ± SD and in dogs with open cervix pyometra, it was mean ± SD. The plasma lactate concentration in dogs with pyometra was higher than in healthy bitches, and there was no influence of patency of the cervix on the concentration of plasma lactate concentrations. Plasma lactate concentrations were similar for animals with open and closed pyometra (3.54 ± 0.52 to 3.64 ± 1.03 mm).

Diabetes mellitus, hypercholesterolemia, and hypertension but not vascular disease per se are associated with persistent platelet activation in vivo. Evidence derived from the study of peripheral arterial disease.

BACKGROUND: Previous studies relating increased thromboxane (TX) biosynthesis to cardiovascular risk factors do not answer the question whether platelet activation is merely a consequence of more prevalent atherosclerotic lesions or reflects the influence of metabolic and hemodynamic disturbances on platelet biochemistry and function. METHODS AND RESULTS: We examined 64 patients with large-vessel peripheral arterial disease and 64 age- and sex-matched control subjects. TXA2 biosynthesis was investigated in relation to cardiovascular risk factors by repeated measurements of the urinary excretion of its major enzymatic metabolite, 11-dehydro-TXB2, by radioimmunoassay. Urinary 11-dehydro-TXB2 was significantly (P = .0001) higher in patients with peripheral arterial disease (57 +/- 26 ng/h) than in control subjects (26 +/- 7 ng/h). Seventy percent of patients had metabolite excretion > 2 SD above the normal mean. However, 11-dehydro-TXB2 excretion was enhanced only in association with cardiovascular risk factors. Multivariate analysis showed that diabetes, hypercholesterolemia, and hypertension were independently related to 11-dehydro-TXB2 excretion. During a median follow-up of 48 months, 8 patients experienced major vascular events. These patients had significantly (P = .001) higher 11-dehydro-TXB2 excretion at baseline than patients who remained event free. CONCLUSIONS: The occurrence of large-vessel peripheral arterial disease per se is not a trigger of platelet activation in vivo. Rather, the rate of TXA2 biosynthesis appears to reflect the influence of coexisting disorders such as diabetes mellitus, hypercholesterolemia, and hypertension on platelet biochemistry and function. Enhanced TXA2 biosynthesis may represent a common link between such diverse risk factors and the thrombotic complications of peripheral arterial disease.

Platelet activation markers in patients with peripheral arterial disease--a prospective comparison of different platelet function tests.

Peripheral vascular disease (PVD) is an indicator of diffuse atherosclerosis and is associated with a greatly increased incidence of coronary heart and cerebrovascular disease. Although several studies have assessed whether in vivo platelet activation takes place in patients with PVD, no data are available comparing different platelet function tests in this patient population. We have compared prospectively four tests for the measurement of in vivo platelet activation (plasma betaTG, plasma PF4, intraplatelet betaTG and urinary excretion of 11-dehydro-TXB2) and one in vitro platelet function test (ADP-induced platelet aggregation) in 63 well-characterized patients with intermittent claudication and in 18 age- and sex-matched healthy volunteers. No statistically significant difference was found between patients and controls for plasma betaTG (20.0 +/- 11.8 vs. 18.8 +/- 9.0 ng/ml, respectively), plasma PF4 (5.2 +/- 2.9 vs. 6.3 +/- 3.5 ng/ml), betaTG/PF4 ratio (4.0 +/- 2.9 vs. 3.6 +/- 1.8), intraplatelet betaTG (4503 +/- 1482 vs. 4059 +/- 1065 ng/ml), and threshold aggregatory concentration of ADP (1.7 +/- 0.72 vs. 1.45 +/- 0.56 microM). Urinary 11-dehydro-TXB2 was instead significantly higher in the PVD group (55.4 +/- 27.5 vs. 26.7 +/- 7.0 ng/h, p <0.001). Our study shows that urinary 11-dehydro-TXB2 is a more sensitive index of in vivo platelet activation than the measurement of either platelet specific proteins or of in vitro platelet aggregation in patients with PVD.

[Fine needle aspiration biopsy of thyroid nodules. Analysis of results obtained using a new method with histological examination of the sample]. / Punción de nódulos tiroídeos con aguja fina. Análisis de los resultados obtenidos utilizando una nueva metódica con examen histológico de la muestra.

Fine needle aspiration biopsy (FNAB) is the most useful preoperative work up method for patients with nodular goiter. We analyze 173 patients that underwent FNAB and were subsequently operated. The sample with FNAB was processed both for cytological and histological study, using methods developed at the hospital. The preoperative diagnosis reached was compared with that obtained with the surgical piece. Results showed that the histological analysis of the FNAB sample improved the sensitivity of the method, when compared with the cytological study, from 68.5% to 87.8% (p < 0.035) without modifying specificity (100% for both methods). It is concluded that FNAB with histological study of the sample has a high diagnostic yield and is exempt of complications.

Detection of asymptomatic deep vein thrombosis by real-time B-mode ultrasonography in hip surgery patients.

The aim of this study was to prospectively evaluate the accuracy of real-time B-mode ultrasonography in the diagnosis of asymptomatic proximal deep vein thrombosis (DVT) in hip surgery patients. Venography was adopted as the gold standard. We studied 100 consecutive patients undergoing hip surgery: 60 patients for hip fracture and 40 patients for elective hip replacement. Bilateral real-time B-mode ultrasonography was performed prior to bilateral venography. The two diagnostic procedures were performed on the same day by different investigators unaware of the assigned prophylatic regimen for DVT. Compressibility of the vein segment was adopted as the single criterion for DVT. Venography was performed and judged by radiologists unaware of the ultrasonography results. In 13 limbs venography was either impossible to perform or not adequate for judgement. Ultrasonography and an adequate venography was obtained in 187 limbs. A venography proven DVT was observed in 49 limbs (26.2%) and a proximal DVT in 21 limbs (11.2%). All the patients were asymptomatic for DVT. The sensitivity and specificity of real time B-mode ultrasonography for proximal DVT were 57% (95% confidence interval: C.I. 36-80) and 99% (C.I. 99-100), respectively and the positive and negative predictive values were 93% (C.I. 73-100) and 95% (C.I. 91-97), respectively. The sensitivity and specificity for overall DVT were 25% (C.I. 11-38) and 99% (C.I. 97-100), respectively and the positive and negative predictive values were 92% (C.I. 73-100) and 79% (C.I. 76-85), respectively. Our data indicate that real-time B-mode ultrasonography for its high specificity could make venography unnecessary in patients with positive results.(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoma adenoideoquístico salival: gradación histológica / Adenoid cystic carcinoma of the salivary gland: histological gradation

Se revisó las características histológicas y clínicas de 27 casos de carcinoma adenoideoquístico (CAQ), de las glándulas salivales mayores y menores. Hubo 13 casos en glándulas salivales mayores (8 submaxilares) y 14 en las menores (4 palatinas). Se reclasificó los tumores en tres grados de malignidad creciente: Grado I: tumores tubulares y cribiformes sin áreas sólidas (14 casos); Grado II: tumores tubulares y cribiformes con menos de 30% de áreas sólidas (8 casos); Grado III: tumores con más de 30% de áreas sólidas (5 casos). Los tumores grado I fueron más pequeños (2,5 cm), predominantemente cribiformes, sin componente sólido, más susceptibles de escisión quirúrgica adecuada y tuvieron un curso clínico prolongado. Los tumores grado II tuvieron 10,62% de áreas sólidas y los grados III, 55,2% de dichas áreas, fueron más grandes (3,7 y 4,8 cm), recurrieron con mayor frecuencia y tuvieron un curso clínico agresivo con muerte del paciente al cabo de 3-4 años. El grado histológico y el tamaño del tumor fueron los parámetros que mejor correlacionaron con la evolución y pronóstico de nuestros pacientes

Double-blind study of dothiepin versus placebo in the treatment of primary fibromyalgia syndrome.

A double-blind study comparing the efficacy and tolerability of dothiepin with that of placebo in the treatment of primary fibromyalgia syndrome was carried out. Dothiepin was shown to improve significantly the condition of patients with primary fibromyalgia syndrome and there was a significant difference between dothiepin and placebo in all the clinical variables measured. Only mild and transient side-effects were reported. Further controlled studies are required to define the effects of dothiepin on fibromyalgia.

Thyroid cancer. Survival in 148 cases followed for 10 years or more.

One hundred forty-eight patients with differentiated carcinoma of the thyroid treated between May 1954 and April 1973 are presented. There were 89 papillary and 59 follicular carcinomas. They were classified according to Woolner criteria. Treatment consisted of surgery I-131 and thyroid hormone. Recurrences occurred in 8.7% of the patients, and lethality at the end of the observation period was 3.3%. The impact of histologic type, extent of the primary, and age of the patient at the time of treatment on prognosis were studied. The data were submitted to statistical analysis. This study revealed that these factors are determinant on prognosis. Best survival rates were observed in patients 40 years of age or younger at the time of treatment, in patients with intrathyroid papillary carcinomas, and in patients with noninvasive follicular carcinomas.

[The treatment of loose diaphyseal fractures of the humeral diaphysis using the hanging cast method]. / Sul trattamento delle fratture diafisarie scomposte della diafisi omerale col metodo gel gesso pendente.

A personal series of loose diaphyseal fractures of the humeral diaphysis, treated with the hanging plaster case method, has been examined in order to assess results. These were positive in most treated cases, with a complication percentage comparable to that obtainable with other methods. Although it is not considered that the usual treatment of reduction and plaster cast thoraco-brachial immobilisation should be abandoned, it is considered that the hanging cast is an equally valid alternative in all cases in which the patient's general condition represents a contraindication to the application of a bulky chest-extended cast.