High MET expression is an adverse prognostic factor in patients with triple-negative breast cancer.

BACKGROUND: The mesenchymal-epithelial transition (MET) pathway is frequently altered in tumours. The purpose of our study was to determine the prognostic value of tumour MET expression levels in patients with triple-negative breast cancer (TNBC), in order to strengthen the rationale for targeted therapy of TNBC using MET inhibitors. METHODS: We determined expression of MET in formalin-fixed paraffin-embedded surgical specimens of TNBC by immunohistochemistry. Recurrence-free and overall survival was analysed with Cox models adjusted for clinical and pathological factors. RESULTS: Immunostaining for MET was classified as high in 89 of 170 (52%) tumours. MET expression was more frequently observed in G3 carcinomas (P=0.02) but was not significantly associated to any of the other clinical or pathological parameters. High MET expression predicted shorter survival of the patients. Multivariate Cox proportional hazards regression analyses identified MET to be an independent prognostic factor for recurrence (adjusted hazard ratio (HR) for recurrence 3.43; 95% confidence interval (CI) 1.65-7.12; P=0.001) and death (adjusted HR for death 3.74; 95% CI 1.65-8.46; P=0.002). CONCLUSION: These results provide further evidence that the MET pathway could be exploited as a target for TNBC.

Effects of muscle type, castration, age, and compensatory growth rate on androgen receptor mRNA expression in bovine skeletal muscle.

The effect of testosterone on sexual dimorphism is evident by differential growth of forelimb and neck muscles in bulls and steers. Divergent hormone sensitivites may account for the differential growth rates of individual muscles. Therefore, the objective of this study was to compare androgen receptor (AR) expression in three different muscles of bulls and steers at various ages and growth rates. Thirty Montbéliard bulls and 30 steers were assigned to four slaughter age groups. Four or five animals of each sex were slaughtered at 4 and 8 mo of age. Animals in the remaining two slaughter groups (12 and 16 mo) were divided into groups of either restricted (R) or ad libitum (AL) access to feed. Five animals of each sex and diet were slaughtered at the end of the restricted intake period at 12 mo of age. To simulate compensatory growth, the remaining animals (R and AL) were allowed ad libitum access to feed until slaughter at 16 mo of age. Total RNA was extracted from samples of semitendinosus (ST), triceps brachii (TB), and splenius (SP) muscles. Androgen receptor mRNA was quantified in 200-ng total RNA preparations using an internally standardized reverse transcription (RT) PCR assay. Data were analyzed using 18S ribosomal RNA concentrations as a covariable. Steers had higher AR mRNA levels per RNA unit than bulls (P < .01). Androgen receptor mRNA levels differed between muscles (P < .05), with lowest expression in the SP. The pattern of AR expression differed (P < .05) for each muscle with increasing age. Between 4 and 12 mo of age, AR mRNA levels increased (P < .05) in SP but remained unchanged in the ST and TB. Feeding regimen had no effect on muscle AR expression, but steers exhibiting compensatory growth had higher AR mRNA levels than AL steers (P < .01) or bulls (P < .01). Our results show that AR expression is muscle-specific and may be modulated by circulating testicular hormones. These data suggest that the regulation of AR expression may be linked to allometric muscle growth patterns in cattle and compensatory gain in steers.

Quantification of myosin heavy chain isoform in porcine muscle using an enzyme-linked immunosorbent assay.

The objective of this study was to develop an indirect enzyme-linked immunosorbent assay (ELISA) to quantify the relative abundance of the myosin heavy chain (MyHC) isoforms in porcine muscle. Longissimus muscle samples were taken from halothane positive (HAL+) and negative (HAL-) pigs and subjected to ELISA using newly generated and commercially available myosin monclonal antibodies (mAbs). Muscle of HAL+ pigs possessed less type I (P <0.01) and IIA (P<0.1), and more type IIB MyHC (P <0.01) than muscle of HAL- pigs. Abundance of IIX MyHC content was negatively correlated (P < 0.0001) to the amount of IIB MyHC in porcine muscle. These data show indirect ELISA can be used to detect genotype differences in muscle MyHC content, and it provides a rapid, sensitive method for determining muscle fiber type in porcine skeletal muscle. Furthermore, these data suggest that the proportion of glycolytic muscle fibers increases at the expense of oxidative fibers.

Uterine androgen receptor mRNA expression in metestrous and anestrous bitches being healthy or suffering from Pyometra.

The importance of androgens for the female reproductive system has been investigated for decades and a number of androgen sensitive processes has now been identified in female reproductive organs. For carnivore species no data were available so far about uterine androgen sensitivity and its regulation. The present study therefore aimed to investigate whether androgen receptors (AR) are present in the dog uterus, whether they are regulated throughout the ovarian cycle and whether pyometra affects their expression rate. Uterine tissue samples were collected from 28 bitches of different ages and various breeds. The samples were grouped according to the stage of estrous cycle (metestrus ME or anestrus AE) and the pathological status of the uterus (i.e. suffering from pyometra or not). Androgen receptor mRNA (AR mRNA) was quantified from 500 ng of total RNA isolated from the tissue samples using an internally standardized reverse transcription polymerase chain reaction (RT-PCR) described previously. The amount of total RNA extractable per g tissue was elevated during pyometra. The successful amplification of the expected 172 bp fragment from canine uterine RNA together with the confirmation of the identity of this fragment by sequence analysis, demonstrates that AR is expressed in this particular tissue. Comparing the expression rates in uteri from bitches during ME or AE being healthy (H) or suffering from pyometra (P), the only significant (p < 0.01) difference was found between H and P uteri during ME with 3.5-fold lower expression rates in P. Although the same seems true for AE bitches, a significant difference could not be demonstrated due to the low number (n = 2) of diseased animals in the AE group. There was no evident effect of the stage of ovarian cycle on uterine AR mRNA levels.

Regional variations of muscle fibre characteristic in m. semitendinosus of growing cattle.

The objectives of the present study were to elaborate an intra-muscular profile of metabolic enzyme equipment, contractile and morphometric features along the longitudinal axis of m. semitendinosus at various ages throughout the growth phase. Thirty-seven male Montbéliard cattle, about half of them castrated, were representatively allocated to various slaughter dates, scheduled at 4, 8, 12 and 16 months of age. Samples were collected from proximal, medial and distal locations of m. semitendinosus. Isocitrate dehydrogenase (aerobic metabolism) and lactate dehydrogenase (anaerobic metabolism) were measured spectrophotometrically. Contractile muscle type was classified by quantification of myosin heavy chain I isoform proportion using the sensitive enzyme-linked immunosorbent assay. Mean muscle fibre area was obtained on histologically-stained cross-sections utilizing an image analysis system. Our results indicate the existence of a regular intra-muscular pattern of muscle fibre traits along the length of m. semitendinosus, with decreasing glycolytic activities and concomitantly an increase in oxidative capacity towards the distal extremity. The metabolic characteristics were in good agreement with decreasing cross-sectional muscle fibre areas and the slow myosin heavy chain I isoform proportion becoming gradually more abundant from proximal to distal regions of the muscle. Moreover, the observed gradient was found to be closely related to age and diminished with advanced physiological maturity. At the final slaughter age (16 months) no differences among the distinct portions were detected, m. semitendinosus was longitudinally homogeneous in all the characteristics studied.

[Extracorporeal shockwave lithotripsy in sedation-analgesia]. / Extrakorporale Stosswellenlithotripsie in Sedoanalgesie.

The aim of the present study was to examine the effects of combined sedation and analgesia during extracorporeal shockwave lithotripsy using the Dornier lithotriptor HM III. We used a combination of a benzodiazepin derivatives with an opioid. We tested the dosage of drugs needed in relation to the length of treatment, the size of the stone and the overall energy output of the lithotriptor. In addition, continuous records were made of the patient's blood pressure and the oxygen saturation in the blood, with and without oxygen insufflation. Our results show that sedation combined with analgesia is a reasonable and useable alternative to general or regional anaesthesia for extracorporeal lithotripsy.