HER-2/neu x aromatase double transgenic mice model: the effects of aromatase overexpression on mammary tumorigenesis

A majority of breast cancers are hormone-responsive, and require estrogen for growth, and respond to hormonal therapy that blocks estrogen receptor action. Breast tumors with low levels of or completely lacking estrogen receptor fail to respond to antiestrogen therapy yet require estrogen for tumor initiation. To address the importance of local estrogen in oncogene-mediated breast tumorigenesis, we have crossed MMTV-aromatase with MMTV-HER2/neu and examined the incidence of breast cancer in double transgenic mice in comparison with parental strains. Double transgenic mice show normal mammary development and express both transgenes at similar levels to that of parental strains. Tumor incidence in double transgenic mice (<5%) decreased compared to HER2/neu mice (>65%). In addition to a significant decrease in tumorigenesis, these mice expressed ER as well as high levels of ERβ along with decreased levels of cyclin D1 and phosphorylated pRb among other changes. Furthermore, experiments using THC (ER-agonist and ERβ-antagonist) clearly demonstrate the critical role of ERβ in HER2/neu-mediated tumorigenesis. These studies provide the first genetic evidence that estrogen receptor, mainly ERβ than ER and its dependent changes play an important role in regulating mammary tumorigenesis. These findings provide further evidence for development and testing of novel therapeutic approaches based on selective regulation of estrogen receptors (ER and β)-dependent actions for the treatment and prevention of breast cancers.     

Myotonic dystrophy protein kinase (DMPK) and its role in the pathogenesis of myotonic dystrophy 1

Myotonic dystrophy 1 (DM1) is an autosomal, dominant inherited, neuromuscular disorder. The DM1 mutation consists in the expansion of an unstable CTG-repeat in the 3'-untranslated region of a gene encoding DMPK (myotonic dystrophy protein kinase). Clinical expression of DM1 is variable, presenting a progressive muscular dystrophy that affects distal muscles more than proximal and is associated with the inability to relax muscles appropriately (myotonia), cataracts, cardiac arrhythmia, testicular atrophy and insulin resistance. DMPK is a Ser/Thr protein kinase homologous to the p21-activated kinases MRCK and ROCK/rho-kinase/ROK. The most abundant isoform of DMPK is an 80 kDa protein mainly expressed in smooth, skeletal and cardiac muscles. Decreased DMPK protein levels may contribute to the pathology of DM1, as revealed by gene target studies. Here we review current understanding of the structural, functional and pathophysiological characteristics of DMPK.     

Within-subject differences in degree of delay discounting as a function of order of presentation of hypothetical cash rewards

Procedural variants in estimating delay discounting (DD) have been shown to yield significant differences in estimated degree of DD as well as variations in individual patterns of choice. For example, a recent study found significantly different degrees of DD between groups assessed using either an ascending or descending order of presentation of the immediately available rewards. The purpose of this study was to test for within-subject effects of order of presentation of the immediate rewards in a DD task. In a single session, college students (N = 29) were asked to complete two DD tasks, one with the immediate rewards presented in ascending order and one in descending order. Consistent with previous results, significantly larger mean area under the discounting curve (AUC) was observed when the descending sequence was used compared to the ascending order of presentation; and the correlation between both measurements was moderate. These results suggest that some DD assessment tasks may be sensitive to contextual variables such as order and range of the reward and delay values.     

Presence of pregnant females of the Gorgona guitarfish,Pseudobatos prahli,in the Mexican tropical Pacific

Five specimens (four females and one male) of the Gorgona guitarfish, Pseudobatos prahli, were sighted during two dives carried out off the western Gulf of Tehuantepec. This document describes the reproductive condition of three adult females captured, two of which had uterine eggs, whereas one had embryos. The sighting and capture of specimens occurred during two Tehuano wind events; the authors, therefore, believe that variations in temperature and dissolved oxygen could have affected P. prahli, leading it to carry out temporal migrations towards coastal shallow waters.     

Chronic oral or intraarticular administration of docosahexaenoic acid reduces nociception and knee edema and improves functional outcomes in a mouse model of Complete Freund’s Adjuvant–induced knee arthritis

Introduction

Clinical and preclinical studies have shown that supplementation with ω-3 polyunsaturated fatty acids (ω-3 PUFAs) reduce joint destruction and inflammation present in rheumatoid arthritis (RA). However, the effects of individual ω-3 PUFAs on chronic arthritic pain have not been evaluated to date. Thus, our aim in this study was to examine whether purified docosahexaenoic acid (DHA, an ω-3 PUFA) reduces spontaneous pain-related behavior and knee edema and improves functional outcomes in a mouse model of knee arthritis.

Methods

Unilateral arthritis was induced by multiple injections of Complete Freund’s Adjuvant (CFA) into the right knee joints of male ICR adult mice. Mice that received CFA injections were then chronically treated from day 15 until day 25 post–initial CFA injection with oral DHA (10, 30 and 100 mg/kg daily) or intraarticular DHA (25 and 50 μg/joint twice weekly). Spontaneous flinching of the injected extremity (considered as spontaneous pain-related behavior), vertical rearing and horizontal exploratory activity (considered as functional outcomes) and knee edema were assessed. To determine whether an endogenous opioid mechanism was involved in the therapeutic effect of DHA, naloxone (NLX, an opioid receptor antagonist, 3 mg/kg subcutaneously) was administered in arthritic mice chronically treated with DHA (30 mg/kg by mouth) at day 25 post–CFA injection.

Results

The intraarticular CFA injections resulted in increasing spontaneous flinching and knee edema of the ipsilateral extremity as well as worsening functional outcomes as time progressed. Chronic administration of DHA, given either orally or intraarticularly, significantly improved horizontal exploratory activity and reduced flinching behavior and knee edema in a dose-dependent manner. Administration of NLX did not reverse the antinociceptive effect of DHA.

Conclusions

To the best of our knowledge, this report is the first to demonstrate DHA’s antinociceptive and anti-inflammatory effects as individual ω-3 PUFAs following sustained systemic and intraarticular administration in a mouse model of CFA-induced knee arthritis. The results suggest that DHA treatment may offer a new therapeutic approach to alleviate inflammation as well as a beneficial effect on pain-related functional disabilities in RA patients.     

Evaluation of different shades to improve dairy cattle well-being in Argentina

 Two tree shades (TS1 and TS2) and an artificial shade structure (AS) were evaluated using black globe temperatures (BGTs) to assess their effectiveness in reducing heat load. The artificial structure consisted of a black woven polypropylene cloth providing 80% shade, mounted on 2.5-m-high eucalyptus posts. The work was carried out at Rafaela Experimental Station, Argentina, during the summer (January and February) 1994. BGTs and floor temperatures were measured in concrete floor holding pens with and without artifical shade. The results showed no difference between TS1, TS2 and AS, their average BGTs being 30.2 (SD 0.58), 29.0 (SD 0.70) and 30.2 (SD 0.74)°C, respectively. BGTs under all three shades were significantly lower (P<0.01) than the average BGT recorded outside: 35.5 (SD 1.12)°C. Average BGTs in holding pens were 32.4 (SD 1.38) and 39.9 (SD 1.91)°C for shaded and non-shaded areas (P<0.01). The corresponding average floor temperatures were 27.8 (SD 0.68)°C and 47.7 (SD 2.13)°C (P<0.01). To assess the effects of shade on animal well-being, afternoon rectal temperatures (RT) and respiratory rate (respirations per minute, RR) of lactating cows were recorded twice a week. Rectal temperatures were significantly higher for non-shaded cows (P<0.01), mean RT being 40.1 (SD 0.65)°C vs 39.3 (SD 0.42)°C for the shaded animals. Corresponding RRs were 78.9 (SD 18.0) and 60.7 (SD 10.6) (P<0.05). It was concluded that: (1) tree and artificial shades produced similar effects, (2) shading the holding pen with an 80% shading cloth was effective in reducing heat load and floor temperatures, and (3) access to shade in our pasture-based system improved animal well-being. Received: 9 September 1996 / Revised: 10 March 1997 / Accepted: 2 July 1997     

3D Imaging Detection of HER2 Based in the Use of Novel Affibody-Quantum Dots Probes and Ratiometric Analysis

Patients with breast cancer (BC) overexpressing HER2 (HER2+) are selected for Trastuzumab treatment, which blocks HER2 and improves cancer prognosis. However, HER2+ diagnosis, by the gold standard, immunohistochemistry, could lead to errors, associated to: a) variability in sample manipulation (thin 2D sections), b) use of subjective algorithms, and c) heterogeneity of HER2 expression within the tissue. Therefore, we explored HER2 3D detection by multiplexed imaging of Affibody-Quantum Dots conjugates (Aff-QD), ratiometric analysis (RMA_FI) and thresholding, using BC multicellular tumor spheroids (BC-MTS) (~120 μm of diameter) as 3D model of BC. HER2+, HER2– and hybrid HER2+/? BC-MTS (mimicking heterogeneous tissue) were incubated simultaneously with two Aff-QD probes (anti-HER2 and negative control (NC), respectively, (1:1)). Confocal XY sections were recorded along the Z distance, and processed by automatized RMA_FI (anti-HER2 Aff-QD/ NC). Quantifying the NC fluorescence allowed to predict the fraction of non-specific accumulation of the anti-HER2 probe within the thick sample, and resolve the specific HER2 level. HER2 was detected up to 30 μm within intact BC-MTS, however, permeabilization improved detection up to 70 μm. Specific HER2 signal was objectively quantified, and HER2 3D-density of 9.2, 48.3 and 30.8% were obtained in HER2?, HER2+ and hybrid HER2+/? permeabilized BC-MTS, respectively. Therefore, by combining the multiplexing capacity of Aff-QD probes and RMA_FI, we overcame the challenge of non-specific probe accumulation in 3D samples with minimal processing, yielding a fast, specific spatial HER2 detection and objective quantification.