ms1, a novel stress-responsive, muscle-specific gene that is up-regulated in the early stages of pressure overload-induced left ventricular hypertrophy

We have identified and characterised a cDNA encoding a novel gene, designated myocyte stress 1 (ms1), that is up-regulated within 1 h in the left ventricle following the application of pressure overload by aortic banding in the rat. The deduced ms1 protein of 317 amino acids contains several putative functional motifs, including a region that is evolutionarily conserved. Distribution analysis indicates that rat ms1 mRNA expression is predominantly expressed in striated muscle and progressively increases in the left ventricle from embryo to adulthood. These findings suggest that rust may be important in striated muscle biology and the development of pressure-induced left ventricular hypertrophy. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

ESC Working Group on Myocardial Function Position Paper: how to study the right ventricle in experimental models

The right ventricle has become an increasing focus in cardiovascular research. In this position paper, we give a brief overview of the specific pathophysiological features of the right ventricle, with particular emphasis on functional and molecular modifications as well as therapeutic strategies in chronic overload, highlighting the differences from the left ventricle. Importantly, we put together recommendations on promising topics of research in the field, experimental study design, and functional evaluation of the right ventricle in experimental models, from non-invasive methodologies to haemodynamic evaluation and ex vivo set-ups.

Phorbol ester, but not ischemic preconditioning, activates protein kinase D in the rat heart

The signal transduction pathways that mediate the cardioprotective effects of ischemic preconditioning remain unclear. Here we have determined the role of a novel kinase, protein kinase D (PKD), in mediating preconditioning in the rat heart. Isolated rat hearts (n=6/group) were subjected to either: (i) 36 min aerobic perfusion (control); (ii) 20 min aerobic perfusion plus 3 min no-flow ischemia, 3 min reperfusion, 5 min no-flow ischemia, 5 min reperfusion (ischemic preconditioning); (iii) 20 min aerobic perfusion plus 200 nmol/l phorbol 12-myristate 13-acetate (PMA) given as a substitute for ischemic preconditioning. The left ventricle then was excised, homogenized and PKD immunoprecipitated from the homogenate. Activity of the purified kinase was determined following bincubation with [γ32P]-ATP±syntide-2, a substrate for PKD. Significant PKD autophosphorylation and syntide-2 phosphorylation occurred in PMA-treated hearts, but not in control or preconditioned hearts. Additional studies confirmed that recovery of LVDP was greater and initiation of ischemic contracture and time-to-peak contracture were less, in ischemic preconditioned hearts compared with controls (P<0.05). Our results suggest that the early events that mediate ischemic preconditioning in the rat heart occur via a PKD-independent mechanism.

Downregulation of cyclin-dependent kinase inhibitors p21 and p27 in pressure-overload hypertrophy

We postulated that the cyclin-dependent kinase inhibitors p21 and p27 could regulate the alterations in growth potential of cardiomyocytes during left ventricular hypertrophy (LVH). LVH was induced in adult rat hearts by aortic constriction (AC) and was monitored at days 0, 1, 3, 7, 14, 21, and 42 postoperation. Relative to sham-operated controls (SH), left ventricle (LV) weight-to-body weight ratio in AC increased progressively with time without significant differences in body weight or right ventricle weight-to-body weight ratio. Atrial natriuretic factor mRNA increased significantly in AC to 287% at day 42 compared with SH (P < 0.05), whereas p21 and p27 mRNA expression in AC rats decreased significantly by 58% (P < 0.03) and 40% (P < 0.05) at day 7, respectively. p21 and p27 protein expression decreased significantly from days 3 to 21 in AC versus SH, concomitant with LV adaptive growth. Immunocytochemistry showed p21 and p27 expression in cardiomyocyte nuclei. Thus downregulation of p21 and p27 may modulate the adaptive growth of cardiomyocytes during pressure overload-induced LVH.

The neural processing of masked speech: Evidence for different mechanisms in the left and right temporal lobes

It has been previously demonstrated that extensive activation in the dorsolateral temporal lobes associated with masking a speech target with a speech masker, consistent with the hypothesis that competition for central auditory processes is an important factor in informational masking. Here, masking from speech and two additional maskers derived from the original speech were investigated. One of these is spectrally rotated speech, which is unintelligible and has a similar (inverted) spectrotemporal profile to speech. The authors also controlled for the possibility of “glimpsing” of the target signal during modulated masking sounds by using speech-modulated noise as a masker in a baseline condition. Functional imaging results reveal that masking speech with speech leads to bilateral superior temporal gyrus (STG) activation relative to a speech-in-noise baseline, while masking speech with spectrally rotated speech leads solely to right STG activation relative to the baseline. This result is discussed in terms of hemispheric asymmetries for speech perception, and interpreted as showing that masking effects can arise through two parallel neural systems, in the left and right temporal lobes. This has implications for the competition for resources caused by speech and rotated speech maskers, and may illuminate some of the mechanisms involved in informational masking.

Superovulation and embryo collection in nulliparous Boer goat does immunized against a recombinant ovine alpha-subunit inhibin

With the purpose of eliciting a superovulatory response, 12 adult nulliparous Boer goat does were actively immunized against a recombinant a-subunit of ovine inhibin (roIHN-alpha; two injections of 100 mg 4 weeks apart). Another 12 control Boer goat does were treated with physiological saline and acted as controls. One year later the immunized animals were boostered by the administration of another dose (100 mg) of the immunogen. Following treatment, blood samples were collected twice weekly for the periods of 16 and 12 weeks, respectively, to monitor the inhibin binding ability with the aid of a radio-tracer binding assay. Throughout the experiment, estrus detection was conducted twice daily with the aid of an aproned intact buck. From the first day after treatment to 48 h after standing estrus, ovarian activity was monitored daily by transrectal ultrasonography. On alternate estrous cycles, does were mated and 6 days later flushed transcervically to recover embryos. All goats treated with the roIHN-alpha produced antibodies reactive to the native bovine inhibin tracer-the titre increasing from 2.9 +/- 0.4 to a maximum of 21.9 +/- 2.9% binding after the second injection. The antibody titre gradually subsided over the next 16 weeks. The booster injection restored an elevated antibody titre (11.7 +/- 0.4%), which was maintained until the end of the sampling period 12 weeks later. In the control goats only trace amounts of antibody were recorded throughout the trial. In the roIHN-alpha-immunized goats the number of follicles reaching a diameter of > 4 mm was 14.6 +/- 1.2 per doe. A positive correlation was recorded between the follicle number and antibody titre (r=0.61; P < 0.01). The number of follicles ovulating per doe (6.9 +/- 0.7) followed the same tendency-however, the proportion decreased with increasing follicle numbers. A relatively weak correlation was recorded between the inhibin binding ability and number of ovulations (r=0.27; P < 0.05). In the control goats the majority (92%) of follicles exceeding 4 mm in diameter ovulated (2.5 +/- 0.1 follicles/doe). Embryo collection proved unsatisfactory (42% versus 39% recovery for immunized and control animals, respectively)-presumably because the uterine lumen of the nulliparous does was too narrow to permit effective flushing. In the group of immunized goats the occurrence of short estrous cycles (< 15 days) recorded was 34% versus only 6% in the controls. Overall, immunization of goats against roIHN-alpha led to an almost six-fold increase in number of ovarian follicles, a three-fold increase in ovulations and, despite the low recovery rate, a more than three-fold increase in ova or embryos recovered. It may be concluded that treatment of female goats with roIHN-alpha leads to an inhibin antibody response, accompanied by enhanced ovarian activity. The response was, however, accompanied by a large proportion of retained follicles and a high incidence of short estrous cycles. These problems need to be further investigated before rendering the method fit for application in embryo transfer programs in goats. (C) 2007 Elsevier B.V. All rights reserved.

Calcium turns sinister in left-right asymmetry

A major challenge in deciphering the development of left-right asymmetry in vertebrates is uncovering how subtle early differences between the left and right sides are translated into robust differences in gene expression. Recently, Raya and colleagues suggested that asymmetric localization of extracellular calcium ions at the node could differentially modulate Notch signalling, resulting in asymmetric expression of the signalling molecule Nodal on the left side of the node. In this article, I examine the implications of this finding and explore their relevance to the evolution of asymmetry in vertebrates and invertebrates.

Cheesemaking with vegetable coagulants - the use of Cynara L. for the production of ovine milk cheeses

This paper reviews the use of plant extracts as vegetable coagulants for cheesemaking. It covers the plants used as sources of coagulants, with a historical overview and particular emphasis on Cynara species. The genus Cynara L., its composition, milk clotting and proteolytic enzymes (cardosins) and their specificity towards peptide linkages are also described. Cheeses produced in the Iberian Peninsula using Cynara L. as coagulant are documented. Cynara L. is still the most used vegetable coagulant in cheesemaking, and also the most investigated. However, much work remains to be done to understand its action during cheese maturation and further characterization.

The neural processing of masked speech: evidence for different mechanisms in the left and right temporal lobes

It has been previously demonstrated that extensive activation in the dorsolateral temporal lobes associated with masking a speech target with a speech masker, consistent with the hypothesis that competition for central auditory processes is an important factor in informational masking. Here, masking from speech and two additional maskers derived from the original speech were investigated. One of these is spectrally rotated speech, which is unintelligible and has a similar (inverted) spectrotemporal profile to speech. The authors also controlled for the possibility of "glimpsing" of the target signal during modulated masking sounds by using speech-modulated noise as a masker in a baseline condition. Functional imaging results reveal that masking speech with speech leads to bilateral superior temporal gyrus (STG) activation relative to a speech-in-noise baseline, while masking speech with spectrally rotated speech leads solely to right STG activation relative to the baseline. This result is discussed in terms of hemispheric asymmetries for speech perception, and interpreted as showing that masking effects can arise through two parallel neural systems, in the left and right temporal lobes. This has implications for the competition for resources caused by speech and rotated speech maskers, and may illuminate some of the mechanisms involved in informational masking.

Hemispace differences in the visual perception of size in left hemi-Parkinson's disease

The visual perception of size in different regions of external space was studied in Parkinson's disease (PD). A group of patients with worse left-sided symptoms (LPD) was compared with a group with worse right-sided symptoms (RPD) and with a group of age-matched controls on judgements of the relative height or width of two rectangles presented in different regions of external space. The relevant dimension of one rectangle (the 'standard') was held constant, while that of the other (the 'variable') was varied in a method of constant stimuli. The point of subjective equality (PSE) of rectangle width or height was obtained by probit analysis as the mean of the resulting psychometric function. When the standard was in left space, the PSE of the LPD group occurred when the variable was smaller, and when the standard was in right space, when the variable was larger. Similarly, when the standard rectangle was presented in upper space, and the variable in lower space, the PSE occurred when the variable was smaller, an effect which was similar in both left and right spaces. In all these experiments, the PSEs for both the controls and the RPD group did not differ significantly, and were close to a physical match, and the slopes of the psychometric functions were steeper in the controls than the patients, though not significantly so. The data suggest that objects appear smaller in the left and upper visual spaces in LPD, probably because of right hemisphere impairment. (C) 2002 Elsevier Science Ltd. All rights reserved.

A left-ear disadvantage for the presentation of irrelevant sound: manipulations of task requirements and changing state

Three experiments attempted to clarify the effect of altering the spatial presentation of irrelevant auditory information. Previous research using serial recall tasks demonstrated a left-ear disadvantage for the presentation of irrelevant sounds (Hadlington, Bridges, & Darby, 2004). Experiments 1 and 2 examined the effects of manipulating the location of irrelevant sound on either a mental arithmetic task (Banbury & Berry, 1998) or a missing-item task (Jones & Macken, 1993; Experiment 4). Experiment 3 altered the amount of change in the irrelevant stream to assess how this affected the level of interference elicited. Two prerequisites appear necessary to produce the left-ear disadvantage; the presence of ordered structural changes in the irrelevant sound and the requirement for serial order processing of the attended information. The existence of a left-ear disadvantage highlights the role of the right hemisphere in the obligatory processing of auditory information. (c) 2006 Published by Elsevier Inc.

Effects of glucose, propionate and splanchnic hormones on neuropeptide mRNA concentrations in the ovine hypothalamus

The capacity for glucose, propionate or hormones of splanchnic origin to influence appetite by directly regulating the expression of neuropeptides in the feeding centres of the hypothalamus of the ruminant is not described. Therefore, our objective was to measure the direct effect of metabolites (glucose and propionate) or hormones [insulin, cholecystokinin (CCK), glucagon-like peptide-1 (GLP-1) and polypeptide YY (PYY)] on hypothalamic mRNA concentrations for neuropeptide Y (NPY), agouti-related peptide (AgRP) and proopiomelanocortin (POMC) following in vitro incubation. Hypothalamic tissue from 4- to 5-month-old lambs was obtained at slaughter and immediately incubated in culture media for 2 h at 36 °C. Treatments included a control Dulbecco’s modified Eagle medium (DMEM) containing 1 mm glucose or DMEM with the following additions: 10 mm glucose, 1 mm propionate, 1 nm insulin, 120 pm GLP-1, 100 pm PYY, 80 pm CCK or 10 mm glucose plus 1 nm insulin. The abundance of mRNA for NPY, AgRP and POMC was measured using quantitative reverse transcriptase PCR. Fisher’s protected LSD test was used to compare changes in relative mRNA concentrations for the hypothalamus incubated in the control media vs. the rest of the treatments. The media containing glucose plus insulin increased POMC mRNA concentration (p < 0.05), but did not affect NPY or AgRP mRNA concentration. There were no effects observed for the other treatments (p > 0.20). Results of the present study are consistent with the concept that effects of propionate on feed intake in ruminants is not mediated through direct effects on the hypothalamus, and that insulin is required for an effect of glucose on hypothalamic POMC expression.