The influence of body size on measurements of overall cardiac function.

The purpose of this study was to determine the best scaling method to account for the effects of body size on measurements of overall cardiac function and subsequently the interpretation of data based on cardiac power output (CPO). CPO was measured at rest (CPO(rest)) and at maximal exercise (CPO(max)) on 88 and 103 healthy but untrained men and women, respectively, over the age range of 20-70 yr. Cardiac reserve (CR) was calculated as CPO(max) - CPO(rest). CPO(rest), CPO(max), and CR were all significantly related to body mass (BM), body surface area (BSA), and lean body mass (LBM). The linear regression model failed to completely normalize these measurements. In contrast, the allometric model produced size-independent values of CPO. Furthermore, all the assumptions associated with the allometric model were achieved. For CPO(rest), mean body size exponents were BM(0.33), BSA(0.60), and LBM(0.47). For CPO(max), the exponents were BM(0.41), BSA(0.81), and LBM(0.71). For CR, mean body size exponents were BM(0.44), BSA(0.87), and LBM(0.79). LBM was identified (from the root-mean-squares errors of the separate regression models) as the best physiological variable (based on its high metabolic activity) to be scaled in the allometric model. Scaling of CPO to LBM(b) (where b is the scaling exponent) dramatically reduced the between-gender differences with only a 7% difference in CPO(rest) and CPO(max) values. In addition, the gender difference in CR was completely removed. To avoid erroneous interpretations and conclusions being made when comparing data between men and women of different ages, the allometric scaling of CPO to LBM(b) would seem crucial.

From CONSENSUS to CHARM--how do ACEI and ARB produce clinical benefits in CHF?

Two decades of research from CONSENSUS to CHARM using modulators of the renin-angiotensin-aldosterone system (RAAS) in chronic heart failure (CHF) patients have shown convincing clinical benefits, but the majority of clinicians prescribing these drugs are still unclear about what mechanisms are responsible for the observed benefits. Of the candidate mechanisms hitherto proposed, there emerges a theme that best fits the spectrum of known factors from pathophysiology of heart failure to how the drugs enhance longevity of patients. This concept can be summarised as follows: after the onset of heart failure, neurohormones are activated resulting in raised levels of angiotensin, aldosterone and catecholamines, which are all known cardiotoxic agents. Cumulatively over time, they are responsible for accelerated cardiomyocyte attrition, manifesting as a faster reduction of cardiac pumping reserve, leading to worsening heart failure, more neurohormonal activation, thus propagating a vicious cycle spiralling towards an earlier fatality. The vicious cycle can be interrupted by dampening the excessive neurohormonal activities, thereby minimising cardiomyocyte losses and preserving cardiac functional reserve for longer. This culminates in maintenance of a reasonable quality of life and enhanced longevity. Such a mechanistic understanding would enable clinicians to have a better perspective on how to apply data from various clinical trials involving these drugs into clinical practice, to optimise and tailor therapy to the individual patient so that each patient can gain maximal benefits.

Growth induced by incremental static stretch in adult rabbit latissimus dorsi muscle.

Incrementally applied static stretch over 3 weeks resulted in a 72 % increase in the weight of the in situ latissimus dorsi muscle in rabbits. True growth rather than tissue oedema was confirmed by increases in the protein content (130 %), the cross-sectional area of the type I fibres (30 %) and the muscle length (i.e. number of sarcomeres in series increased 25 %). Despite an increase in the proportion of fibres staining positive for the enzyme succinate dehydrogenase (SDH), the myosin ATPase stain showed no appreciable fibre type transformation. While total power output in the stretched muscle was unchanged, its maximum mass specific power output, as determined by oscillatory work loops, was decreased by 50 %. The cross-sectional area that was occupied by connective tissue increased from 15 to 19 % in the stretched muscles, with a concomitant increase in passive energy dissipation. Some incrementally stretched muscles were then allowed an additional 3 weeks of maintained stretch to determine whether the adaptive changes would be preserved or reversed. Previous gains in muscle weight, length and area of type I fibres all remained. In contrast, the connective tissue content and the passive properties returned to control values during this period.

Mechanical properties of rabbit latissimus dorsi muscle after stretch and/or electrical stimulation.

The work loop technique was used to measure the mechanical performance in situ of the latissimus dorsi (LD) muscles of rabbits maintained under fentanyl anesthesia. After 3 wk of incrementally applied stretch the LD muscles were 36% heavier, but absolute power output (195 mW/muscle) was not significantly changed relative to that of external control muscle (206 mW). In contrast, continuous 10-Hz electrical stimulation reduced power output per kilogram of muscle >75% after 3 or 6 wk and muscle mass by 32% after 6 wk. When combined, stretch and 10-Hz electrical stimulation preserved or increased the mass of the treated muscles but failed to prevent an 80% loss in maximum muscle power. However, this combined treatment increased fatigue resistance to a greater degree than electrical stimulation alone. These stretched/stimulated muscles, therefore, are more suitable for cardiomyoplasty. Nonetheless, further work will be necessary to find an ideal training program for this surgical procedure.

Isometric and isotonic muscle properties as determinants of work loop power output.

The power output of rabbit latissimus dorsi muscle was calculated under isotonic conditions and during oscillatory work. Isotonic shortening studies yielded a maximum power output of 120 W . kg-1 at a P/P0 of 0.4 compared to a maximum power output of 32 W . kg-1 obtained using the work loop technique. This difference can largely be explained by comparing actual work loops with those constructed using force velocity (P/V) and isometric data. At low cycle frequencies, work loop power output is quite close to that predicted from P/V and isometric data. However, at higher frequencies other dynamic muscle properties appear to exert a more marked effect.

The induction of c-fos and c-jun in the stretched latissimus dorsi muscle of the rabbit: responses to duration, degree and re-application of the stretch stimulus.

The mRNA levels of the proto-oncogenes c-fos and c-jun were measured in the rabbit latissimus dorsi (LD) muscle in response to the application of various stretch regimes in vivo. It was shown that it was necessary for the stretch to be applied continuously over 1 h in order to achieve full induction of c-fos and c-jun mRNA at 1 h. In addition, a correlation was demonstrated between the degree of stretch imposed on the LD and the induced levels of c-fos and c-jun mRNA. Moreover, a second induction of these genes occurred when the LD was subjected to a second stretch stimulus. The magnitude and time course of the second response depended on the precise timing of the second stimulus in relation to the initial induction.

The temporal and cellular expression of c-fos and c-jun in mechanically stimulated rabbit latissimus dorsi muscle.

The levels of c-fos and c-jun mRNA were measured by reverse transcription PCR in the rabbit latissimus dorsi muscle following three separate training regimes, i.e. passive stretch, 10 Hz electrical stimulation or a combination of the two. Both c-fos and c-jun mRNA expression peaked at around 1 h after imposing stretch and at around 4.5-6 h after the initiation of electrical stimulation. The combined stretch/electrical stimulation regime induced biphasic expression of both c-fos and c-jun mRNA, with peaks coinciding temporally with those for the individual regimes. Immunostaining with anti-Fos and anti-Jun antibodies revealed the accumulation of these proteins in both myofibre and interstitial cell nuclei following passive stretch. In contrast, following electrical stimulation the localization of immunoreactive c-Fos and c-Jun proteins was predominantly in interstitial cell nuclei. c-Fos and c-Jun immunoreactivity was also clearly colocalized in a proportion of myonuclei from stretched muscle. These findings suggest that the rapid induction of c-fos and c-jun is an early event in response to mechanical stretch and might trigger [via activator protein-1 (AP-1) transcriptional factors] events leading to muscle fibre hypertrophy. However, the involvement of AP-1 in inducing the phenotypic changes in muscle fibres as a result of electrical stimulation appears less clear.

Muscle growth in response to mechanical stimuli.

The relative merits of the separate and combined uses of stretch and electrical stimulation at 10 Hz in influencing the rates of protein synthesis in vivo, proteolysis, and the growth of the extensor digitorum longus muscle have been investigated after 3 days in the rabbit. Continuous electrical stimulation failed to change muscle protein turnover or growth. Static stretch caused significant adaptive growth, with increases in c-fos, c-jun, and insulin-like growth factor I (IGF-I; 12-fold) mRNA levels, and protein (19%), RNA (128%), and DNA (45%) contents. Both the fractional (138%) and total (191%) rates of protein synthesis increased with stretch, correlating with increased ribosomal capacities. Combining stretch and electrical stimulation increased the mRNA concentration of IGF-I (40-fold). The adaptive growth was greater (35%), with massive increases in the nucleic acids (185 and 300%), ribosomal capacities (230%), and the rates of protein synthesis (345 and 450%). Large increases (i.e., 200-400%) in cathepsins B and L and dipeptidyl aminopeptidase I activities during stretch, with or without stimulation, suggest a role for these enzymes in tissue remodeling during muscle hypertrophy.

The mechanical properties of fast and slow skeletal muscles of the mouse in relation to their locomotory function.

The mechanical properties of soleus and extensor digitorum longus (EDL) muscles from the mouse were studied using the work loop technique. Under optimum conditions, the EDL produced a maximum mean power output of 107 W kg-1 at a cycle frequency of 10 Hz. In comparison, the maximum mean power output of the soleus was 34 W kg-1 at 5 Hz cycle frequency. Video analysis of mice determined the stride frequency range to be from 2.87 Hz at a walk to 8.23 Hz at a flat-out gallop, with the trot-to-gallop transition occurring at 5.89 Hz. In vivo EDL electromyogram (EMG) activity is recorded primarily during shortening and the muscle operates in a power-generating mode. The soleus is close to isometric when EMG activity is recorded, but mechanical activity persists into the shortening phase. Both muscles are likely to operate over cycle frequency ranges just below, or at, those yielding maximal power. Soleus and EDL produced maximal power output in vitro when operating at mean sarcomere lengths of 2.58 microns and 2.71 microns respectively. These lengths are slightly above the plateau of the length-force curve predicted for rat leg muscle (2.3-2.5 microns). The sarcomere length ranges used in vivo by the soleus and EDL were determined, by fixing muscles in the extreme active positions predicted from video and cine analysis, to be 2.28-2.57 microns and 2.49-2.88 microns respectively. These ranges are both close to those shown to yield maximum power output in vitro and to the plateau of the sarcomere length-force curve.

Thyroid-induced changes in the growth of the liver, kidney, and diaphragm of neonatal rats.

Eu-, hypo- and hyper-thyroid rats were studied 12 days postpartum. Hypothyroidism was induced by administering propylthiouracil (PTU) via the mother's drinking water between late gestation and throughout lactation. This procedure effectively blocked the normal early postnatal surge of T3 and T4. In contrast, hyperthyroidism was induced in the young pups by daily injections of T4 from day 3 postpartum. The effects of these experimental manipulations of thyroid status on the rates of protein turnover and growth of the liver, kidney, and diaphragm were studied and compared with measurements made on appropriate euthyroid control tissues. Tissue rates of protein synthesis were decreased in response to hypothyroidism with consequent growth retardation of all three tissues and the whole animal. In contrast, the three body tissues responded very differently to the induction of hyperthyroidism. Hepatic rates of protein synthesis and growth were completely unaffected by thyroid excess. The response of the diaphragm was essentially the reverse of that seen with hypothyroidism, i.e., the enhanced rates of protein synthesis and protein degradation leading to muscle hypertrophy. The rates of protein turnover in the kidney were also increased, but unlike the diaphragm the net result was renal atrophy. Clearly, thyroid hormones influence the normal rapid growth of the neonate and its individual tissues. However, beyond a certain concentration the threshold of responsiveness to these hormones seems to vary between individual tissues.

The fibre type composition of the rabbit latissimus dorsi muscle.

The fibre type distribution has been mapped in the latissimus dorsi muscle of the Dutch rabbit. Using the myosin ATPase stain, a distinct border was found to run in a cranial to caudal direction, which effectively divided the muscle into 2 segments of different fibre type proportions. Although both segments contained mostly fast twitch fibres, the medial areas were found to contain approximately 10-20% slow (i.e. type I) fibres while the lateral portions contained very few, if any, slow fibres. Significantly fewer type IIa fibres were also found in the lateral areas of the muscle. These histochemical findings were confirmed by the use of the reverse transcriptase polymerase chain reaction, which demonstrated that more messenger RNA of the slow myosin heavy chain was found in the medial regions compared with the lateral segment. These results demonstrate the importance of choosing well defined sampling sites when evaluating regimes designed to transform this heterogeneous muscle for use in subsequent myoplasty procedures.

The influence of thyroid hormones on the growth of the atria and ventricles of the heart in immature rats.

The normal plasma concentrations of tri-iodothyronine (T3) and thyroxine (T4) increase approximately six- and fourfold respectively between the end of gestation and weaning in the rat. This early postnatal surge of thyroid hormones was experimentally modified to produce either a state of hypo- or hyperthyroidism. The growth and rates of protein turnover in the atria and ventricles of the heart were studied, 12 and 20 days postpartum, both as a function of age and of changing thyroid status. Neonatal hypothyroidism was induced by adding propylthiouracil to the mothers' drinking water late in gestation and throughout lactation. Hyperthyroidism was achieved by giving the suckling pups daily injections of T4 from day 3 postpartum onwards. Between 12 and 20 days the weight and protein mass of the combined ventricles of the euthyroid animals approximately doubled, along with substantial increases (50%) in the RNA and DNA contents. Over this same 8 days, growth in the combined atria was much slower. During the same period, hypothyroidism significantly retarded the growth of these immature rats and their atria and ventricles. Both the rates of protein synthesis and protein degradation were decreased in the atria and ventricles. In contrast, hyperthyroidism significantly increased growth in both types of cardiac tissue, this being more pronounced in the atria than in the ventricles between 12 and 20 days. The rates of protein synthesis were increased accordingly, principally by increases in the ribosomal activities. In conclusion, thyroid hormones clearly influence the early postnatal growth of the atria and ventricles of the heart in the rat.

The influence of thyroid hormones on the growth of the lungs in perinatal rats.

The growth and rates of protein turnover in the perinatal lung have been studied in the rat during normal development between late gestation and weaning, and after altering their thyroid status. The aim was to establish what influence thyroid hormones have on the early stages of growth in the lungs. Perinatal hypothyroidism was induced by administering propylthiouracil (PTU) via the mothers' drinking water from late gestation and throughout lactation. A precocious and elevated surge of thyroid hormones was induced by daily injections of T4 from day 3 postpartum onwards. Hypothyroidism in the neonate, but not the fetus, significantly retarded the growth of the animal and its lungs. This was attributable to a decrease in both the pulmonary rates of protein synthesis and protein degradation; the effect on the former rate exceeding that on the latter. Neonatal hyperthyroidism did not significantly alter protein turnover or the growth of the lungs, compared with euthyroid control tissues. This contrasts with the accelerated growth of some other body tissues in the presence of excess thyroid hormones.

The effects of established diabetes on the growth of the fetal liver and skin in the rat.

1. The effects of uncontrolled maternal diabetes on the growth of the whole rat fetus and its liver and skin were studied over the last 4 days of gestation. 2. Smaller fetuses, with growth-retarded livers and skins, were consistently found between 18 and 21 days in the diabetic pregnancies. 3. The smaller diabetic livers and skins (i.e. combined epidermis and dermis) possessed lower protein, RNA and DNA contents, compared with the same fetal tissues from normal pregnancies. 4. The growth retardation of the livers in diabetic fetuses was attributed to fewer normally sized hepatic cells. 5. Whilst hepatic rates of protein synthesis (measured in vivo) remained largely unchanged, these were actually increased in the skin, compared with normal control tissues. 6. These findings point to an elevated rate of protein degradation in both diabetic tissues as the most likely cause of their retarded growth.

The influence of age and chronic restricted feeding on protein synthesis in the small intestine of the rat.

Rates of protein synthesis (measured in vivo) and growth of the small intestine were studied as a function of age in ad libitum fed (control) and chronic dietary-restricted rats. At weaning, the fractional rates of synthesis in the mucosal and muscularis externa and serosal layers of the small intestine of control animals were similarly high (90-100% per day). Although these rates subsequently declined with age in the muscularis externa and serosa, they remained constant in the mucosa. Restricted feeding (50% reduced intake), when imposed from weaning onwards, significantly extends the maximum life span of rodents. However, the change in nutritional status slows the accumulation of protein, RNA, and DNA in both layers of the small intestine. Although underfeeding did not prevent the age-related fall in muscularis externa and serosal protein synthesis, significantly higher rates (both fractional and per ribosome) were found when compared age for age with controls. Mucosal fractional synthetic rates were similarly increased by the reduced food intake. These changes in protein turnover in the small intestine are consistent with the higher rates of whole body turnover previously observed in chronically underfed rats.