Hypoxic pulmonary vasoconstriction (HPV) is an important mechanism for matching the ventilation/perfusion ratio in the lung, but the signal transduction pathway through which hypoxia induces vasoconstriction remains unclear. We hypothesized that the decrease in K+ current induced by hypoxia is a key mechanism for HPV, and examined the effects of the substances which are expected to accumulate during hypoxia on the activity of large conductance Ca2+-activated K+ (BKCa) channels. Pulmonary and ear arterial smooth muscle cells were isolated from the rabbit using enzymatic digestion, and large conductance Ca2+-activated K+ current (IBK,Ca) was recorded in symmetrical K+ concentrations using the inside-out mode of the patch-clamp technique. Increasing the Na+ concentration on the intracellular side suppressed IBK,Cadose dependently: 4·6, 20·9, 35·5 and 44·6 % reduction with 4, 8, 12 and 16 mM Na+, respectively. Mg2+ also reduced IBK,Ca, and the maximum reduction was obtained at 0·5 mM. Lactate, adenosine, ADP and ATP did not significantly affect IBK,Ca. There was no difference between pulmonary and ear arterial smooth muscle cells in their response to the above substances; this finding rules out modulation of BKCa channels by the various factors thought to accumulate during hypoxia as a major mechanism involved in the decrease in the K+ conductance of pulmonary arteries in hypoxia.

The intracellular mechanisms responsible for inhibition of osteoclast activity are of significant interest in the search for more effective ways of managing bone diseases associated with enhanced bone resorption. Previous studies have suggested that the protein kinase C (PKC) pathway is an important inhibitory second messenger in osteoclasts. We, therefore, investigated the effects of the synthetic peptide fragments, PKC(530-558) and (19-36), which correspond to parts of the catalytic and regulatory domains of PKC, on the activity of isolated osteoclasts. These fragments have been shown to activate and inhibit PKC, respectively, in biochemical studies employing isolated rat brain PKC, but have rarely been employed in studies of cellular activity. PKC(19-36), an enzyme inhibitor (PKC-I), had no effect by itself on osteoclastic bone resorption. However, PKC(530-558), a PKC activator (PKC-A), caused a dose-responsive inhibition of bone resorption, which was accompanied by a rapid and distinctive change in osteoclast morphology. This effect was reversible: (a) upon removal of PKC-A, (b) upon continuous exposure to this fragment for more than 36 h, or (c) in the presence of PKC-I. In conclusion, a short synthetic peptide fragment of PKC (PKC-A) significantly inhibits osteoclastic bone resorption; this, together with the fact that the inhibitory effect is abolished in the presence of PKC-I, provides further evidence for an important physiological role for the PKC pathway in the regulation of osteoclast activity. Selective activation of this pathway may have important therapeutic implications for the management of bone diseases associated with enhanced resorption.

The acinar cells of the ferret parotid gland are supplied with parasympathetic nerve fibres containing vasoactive intestinal peptide (VIP) and substance P. In this study intracarotid infusions of the two peptides (0·5-1·0 µg kg-1 min-1 of each for 40 min) in the pentobarbitone-anaesthetized ferret, treated with atropine and adrenoceptor antagonists, induced a loss of acinar secretory granules from this gland, by 32 % in response to VIP and by 46 % in response to substance P. Stimulation of the parasympathetic auriculo-temporal nerve (40 Hz, in the presence of adrenoceptor antagonists) caused a larger loss of acinar granules from the gland than stimulation of the sympathetic superior cervical ganglion (intermittently, 50 Hz for 1 s every 10th second, in the presence of atropine) over 40 min (52 % versus 10 %). A 27 % granular loss in response to parasympathetic stimulation followed upon atropinization. The parasympathetic response was not further diminished by the tachykinin antagonist Spantide ((D-Arg1, D-Pro2, D-Trp7,9, Leu)-substance P). Thus, despite the large exocytotic response to the infusion of substance P, the parasympathetic non-adrenergic, non-cholinergic secretion of storage granules seemed, under the present experimental conditions, to occur independently of the action of substance P.

Electrophysiological investigation of neurones in sacral segments of the spinal cord was performed in [alpha]-chloralose-anaesthetized cats in order to establish whether at least some ascending tract neurones could diverge to three different centres located in the brainstem, the cerebellum or the spinal cord. Recordings of antidromic action potentials from cells in S1 and S2 segments were taken following stimulation of the contralateral gigantocellular nucleus, contralateral restiform body and ipsi- and contralateral grey matter of the C6 spinal segment. Antidromic responses allowed identification of several types of neurones that differed in their pattern of supraspinal or propriospinal projections. In eighteen out of a total of sixty-three neurones triple projections to all the above structures were found. In the majority of cells investigated their axons divided into two branches ascending both ipsi- and contralaterally in the lateral funiculi of the spinal cord. Their cell bodies were distributed in laminae VII-VIII except for a minor group of neurones that projected to the C6 segment only, which were located in laminae V-VI. The latter group also displayed lower values of axonal conduction velocities. Comparison of conduction velocities in proximal and distal parts of axons revealed significant slowing in most, raising the possibility that additional collaterals were present to other spinal or supraspinal centres. Dual and triple projections from most cells in this study suggest that such a divergence may be a more common feature of ascending tract neurones than has been reported before.

Regional differences in action potential characteristics and membrane currents were investigated in subendocardial, midmyocardial and subepicardial myocytes isolated from the left ventricular free wall of guinea-pig hearts. Action potential duration (APD) was dependent on the region of origin of the myocytes (P < 0·01, ANOVA). Mean action potential duration at 90 % repolarization (APD90) was 237 ± 8 ms in subendocardial (n = 30 myocytes), 251 ± 7 ms in midmyocardial (n = 30) and 204 ± 7 ms in subepicardial myocytes (n = 36). L-type calcium current (ICa) density and background potassium current (IK1) density were similar in the three regions studied. Delayed rectifier current (IK) was measured as deactivating tail current, elicited on repolarization back to -45 mV after 2 s step depolarizations to test potentials ranging from -10 to +80 mV. Mean IK density (after a step to +80 mV) was larger in subepicardial myocytes (1·59 ± 0·16 pA pF-1, n = 16) than in either subendocardial (1·16 ± 0·12 pA pF-1, n = 17) or midmyocardial (1·13 ± 0·11 pA pF-1, n = 21) myocytes (P < 0·05, ANOVA). The La3+-insensitive current (IKs) elicited on repolarization back to -45 mV after a 250 ms step depolarization to +60 mV was similar in the three regions studied. The La3+-sensitive tail current, (IKr) was greater in subepicardial (0·50 ± 0·04 pA pF-1, n = 11) than in subendocardial (0·25 ± 0·05 pA pF-1, n = 9) or in midmyocardial myocytes (0·38 ± 0·05 pA pF-1, n = 11, P < 0·05, ANOVA). The contribution of a Na+ background current to regional differences in APD was assessed by application of 0·1 µM tetrodotoxin (TTX). TTX-induced shortening of APD90 was greater in subendocardial myocytes (35·7 ± 7·1 %, n = 11) than in midmyocardial (15·7 ± 3·8 %, n = 10) and subepicardial (20·2 ± 4·3 %, n = 11) myocytes (P < 0·05, ANOVA). Regional differences in action potential characteristics between subendocardial, midmyocardial, and subepicardial myocytes isolated from guinea-pig left ventricle are attributable, at least in part, to differences in IK and Na+-dependent currents.

The passive and active electrical properties of left ventricular myocardium were measured, using conducted action potentials and current clamp of isolated myocytes. The objective was to quantify changes of intracellular resistivity, Ri, during hypertrophic growth and the simultaneous imposition of cellular hypoxia. Ri was estimated from the time course of the rising phase of a conducted action potential using a solution of the two-dimensional cable equation. The thoracic aorta of guinea-pigs was constricted to induce left ventricular hypertrophy (LVH) and myocardium used 50 and 150 days post-operation. Conduction velocity increased in the earlier stage of LVH and declined in the later stage, compared with age-matched controls. Hypoxia reduced conduction velocity in all experimental groups. Ri increased only in the later stage of hypertrophy (253 ± 39 [Omega] cm to 544 ± 130 [Omega] cm) and was additionally increased by hypoxia in all groups (e.g. control myocardium 252 ± 39 [Omega] cm to 506 ± 170 [Omega] cm). The magnitude of the increase of Ri in hypertrophied, hypoxic myocardium can create conditions required to generate re-entrant arrhythmias.

We studied the development of nutritional iron deficiency 0, 10, 20, 30 and 40 days after the intake of a semisynthetic diet lacking iron (diet 0) and the possible interactions with calcium, phosphorus and magnesium in both control rats and rats after 40 days of iron deficiency. During this period, iron deficiency was found to produce stress in the rats, as evidenced by high levels of cortisol in the serum. High levels of parathyroid hormone (PTH) were also found. There was a considerable increase in the absorption of calcium, phosphorus and magnesium, but the phosphorus and magnesium balance decreased and that of calcium remained practically unchanged, although there was an increase in calcium urinary elimination. Despite the noticeable degree of bone demineralization, which was evident in the femur, serum levels of calcium, phosphorus and magnesium remained constant. The present study shows that severe nutritional ferropenic anaemia provokes significant alterations in the metabolism of calcium, phosphorus and magnesium. We conclude that these alterations should be taken into account in the treatment of this pathology, given its prevalence and the fact that it may exacerbate other pathologies, particularly those related to the metabolism of calcium and phosphorus.

Four adult Corriedale sheep were used in an experiment divided into three parts. In part 1 a primed continuous infusion of [6,6-2H2]glucose was infused for 7 h. The first 3 h was the control period, from 3 to 7 h glucose-dependent insulinotropic polypeptide (GIP) was infused, and from 5 to 7 h somatostatin was infused. Part 2 of the experiment was the same as for part 1 except that insulin was infused between 3 h and 7 h and GIP was infused between 5 and 7 h. Coincident with the insulin infusion, normal glucose was also infused at a variable rate in order to keep the plasma glucose at basal levels. In part 3 of the experiment [6,6-2H2]glucose was infused for 5 h and somatostatin was infused between 3 and 5 h. Measurements of glucose turnover were made in the last 40 min of the control, GIP only, insulin only, somatostatin only, GIP plus somatostatin and GIP plus insulin infusion periods. Plasma insulin levels were reduced to the limit of detection by the somatostatin infusion; under such conditions whole-body glucose uptake should be entirely non-insulin-mediated (NIMGU). Expressing glucose disposal as glucose metabolic clearance rate demonstrated that elevated, but still physiological GIP levels had no effect on NIMGU but significantly increased insulin-mediated glucose uptake when plasma insulin levels were similar to levels typically observed after a meal. These results indicate that in sheep, GIP may enhance insulin action with respect to glucose disposal following a meal, but has no effect on glucose disposal pathways not responsive to insulin.

Newborn pigs rely essentially on shivering thermogenesis in the cold. In order to understand the rapid postnatal enhancement of thermogenic capacities in piglets, the oxygen and nutrient uptake of hindquarters was measured in vivo in 1- (n = 6) and 5-day-old (n = 6) animals at thermal neutrality and during cold exposure. The hindquarters were considered to represent a skeletal muscle compartment. Indirect calorimetry and arterio-venous techniques were used. The cold challenge (23¡C at 1 day old and 15¡C at 5 days old for 90 min) induced a similar increase (+90 %) in regulatory heat production at both ages. Hindquarters blood flow was higher at 5 days than 1 day old at thermal neutrality (26 ± 3 vs. 17 ± 1 ml min-1 (100 g hindquarters)-1) and its increase in the cold was much more marked (+65 % at 5 days old vs. +25 % at 1 day old). Oxygen extraction by the hindquarters rose from 30-35 % at thermal neutrality to 65-70 % in the cold at both ages. The calculated contribution of skeletal muscle to total oxygen consumption averaged 34-40 % at thermal neutrality and 50-64 % in the cold and skeletal muscle was the major contributor to regulatory thermogenesis. Based on hindquarters glucose uptake and lactate release, carbohydrate appeared to be an important fuel for shivering. However, net uptake of fatty acids increased progressively during cold exposure at 5 days old. The enhancement in muscular blood supply and fatty acid utilization during shivering is probably related to the postnatal improvement in the thermoregulatory response of the piglet.

The effects of exogenous arginine vasotocin (AVT), isotocin (IT) and melatonin on the blood pressure of conscious Atlantic cod (Gadus morhua) were examined. Ventral aortic blood pressure (PVA) was recorded from a cannula inserted into the afferent branchial artery of the third gill arch. Dorsal aortic blood pressure (PDA) was recorded via a cannula implanted into the efferent branchial artery in the same gill arch. Each fish received two doses (10 and 50 ng kg-1) of AVT, IT and melatonin. AVT was also administered in combination with melatonin. Injection of 10 ng kg-1 AVT produced significant hypertension, especially in animals injected during the daytime. In contrast, the same dose of IT induced no significant change in either PVA or PDA. Administration of 10 ng kg-1 melatonin at night caused a long-lasting decrease in both parameters. Melatonin also inhibited the increase in blood pressure elicited by AVT. These results indicate that AVT, but not IT, is vasopressor in the cod. The effects of combined administration of melatonin and AVT show the antihypertensive action of melatonin in AVT-induced hypertension.

The present study was planned to investigate the effect of left ventricular pressure and inotropic state on coronary arterial inflow in systole in the anaesthetized dog. A wide range of left ventricular systolic pressures, including the physiological range, were studied. Experiments were done under conditions of maximal vasodilatation and low perfusion pressure in order to avoid vascular autoregulative interference and to keep the microvascular pressure within the normal range. In five anaesthetized dogs, perfused with extracorporeal circulation system, ventricular volume was changed from 20 to 50 ml in steps of 10 ml by filling an intraventricular latex balloon, and the related changes in left ventricular pressure and coronary flow were measured. The volume was then extended to 70 ml to obtain an overstretch which induced a transient decrease in cardiac contractility. During the period of low cardiac contractility the volume was brought back to 20 ml in steps of 10 ml. Systolic ventricular pressure changed with volume but was lower during the period of low contractility. For systolic pressures below 100 mmHg there was no significant relationship between pressure and coronary systolic flow, but the relationship shifted to higher flows during low contractility. For systolic pressures above 100 mmHg systolic coronary flow decreased significantly when systolic pressure increased. In this case the slopes of the relationships were not significantly different before and after the reduction in contractility. These findings suggest that for systolic pressures less than 100 mmHg (i.e. below the physiological range) the shielding effect of the contracting ventricle prevents the ventricular pressure from being transmitted in the myocardial wall. When systolic pressure exceeds 100 mmHg the shielding effect is overcome and the amplitude of the systolic flow reduction varies with ventricular pressure.

The mechanism by which pulmonary blood flow increases and pulmonary vascular resistance decreases after birth is not fully understood. The aim of this study was to simulate the decrease in lung volume caused by the onset of air-breathing at birth and determine whether it can duplicate the changes in pulmonary blood flow and vascular resistance that occur at this time. In chronically catheterized fetal sheep near term (145 days of gestation), fetal pulmonary arterial blood flow was measured, using coloured microspheres, before and after fetal lung liquid volumes were reduced from 52·2 ± 2·7 to 21·2 ± 1·6 ml kg-1. During the 30 min period following the reduction in lung liquid volume, the pulmonary-to-systemic arterial pressure difference decreased from 6·8 ± 1·2 mmHg (pulmonary > systemic) to 1·6 ± 0·5 mmHg. Reducing the volume of fetal lung liquid increased pulmonary blood flow from 59·1 ± 10·5 to 204·2 ± 40·4 ml min-1 (100 g tissue)-1 and reduced pulmonary vascular resistance from 0·53 ± 0·20 to 0·14 ± 0·04 mmHg min ml-1 (100 g tissue)-1. We conclude that a reduction in fetal lung liquid volume, which simulates the reduction in lung volume that occurs at birth, causes a 3- to 4-fold increase in pulmonary blood flow and a reduction in pulmonary vascular resistance of a similar magnitude. Thus, the reduction in lung volume associated with the lung changing from a liquid- to an air-filled organ, may partly account for the increase in pulmonary blood flow and decrease in pulmonary vascular resistance at birth.

The present study examined the hypothesis that the antagonistic ankle dorsiflexor coactivation level during maximum isometric voluntary plantarflexion (MVC) is a function of ankle angle. Six male subjects generated plantarflexion and dorsiflexion MVC trials at ankle angles of -15 deg (dorsiflexed direction), 0 deg (neutral position), +15 deg (plantarflexed direction) and +30 deg having the knee flexed at an angle of 90 deg. In all contractions surface EMG measurements were taken from tibialis anterior and soleus which were considered representative muscles of all dorsiflexors and plantarflexors, respectively. Antagonistic dorsiflexor coactivation was expressed as normalized EMG and moment. Calculations of the antagonistic dorsiflexor moment were based on the tibialis anterior EMG-dorsiflexor moment relationship from contractions at 50, 40, 30, 20 and 10 % of the dorsiflexion MVC moment. In both legs dorsiflexor coactivation level followed an open U-shaped pattern as a function of ankle angle. Differences of 9 and 14 % (P < 0·05) were found in the measured net plantarflexion MVC moment between legs at ankle angles of -15 and +30 deg, respectively. No difference (P > 0·05) was found in the calf circumference between legs. Differences were found in the antagonistic dorsiflexor coactivation between legs at ankle angles of -15 and +30 deg. In the weaker leg the antagonistic EMG measurements were higher by 100 and 45 % (P < 0·01) and the estimated antagonistic moments were higher by 70 and 43 % (P < 0·01) compared with the weaker leg at -15 and +30 deg, respectively. This finding was associated with a decreased range of motion (ROM) in the weaker leg (14 %, P < 0·01), such that no difference (P > 0·05) was found in dorsiflexor antagonistic coactivation between legs at end-range ankle angles. The findings of the study (i) have to be taken into consideration when estimating musculoskeletal loads in the lower extremity, (ii) imply that stretching training can result in a stronger plantarflexion at end-range ankle angles through inhibition of the dorsiflexors, and (iii) imply a neural drive inadequacy during a plantarflexion MVC at end-range angles.

The control of self-initiated falls from different heights was studied. The objective of the study was to investigate in a quantitative manner the modulation of EMG timing (i.e. onset from take-off and duration from onset to touch-down) and amplitude (before and after foot contact) as a function of fall height. The muscles studied were m. soleus and m. tibialis anterior. Kinematic (ankle joint angle) and kinetic (ground reaction force) variables were also measured. Six subjects took part in the experiments that consisted of ten landings from each of five heights (0·2, 0·4, 0·6, 0·8 and 1 m) onto a force platform. We found a consistent pattern of co-contraction before and after touch-down across the fall heights studied. In both muscles, the onset of pre-landing EMG activity occurred at a longer latency following take-off when landing from greater heights. The absolute EMG duration was affected to a lesser extent by increasing fall height. These findings suggest that the onset of muscle activity of the muscles studied prior to foot contact is timed relative to the expected time of foot contact. Pre- and post-landing EMG amplitude tended to increase with height. Despite a doubling in the magnitude of ground reaction force, the amplitude of ankle joint rotation caused by the impact remained constant across heights. These findings suggest that the observed pattern of co-contraction is responsible for increasing ankle joint stiffness as fall height is increased. The attainment of an appropriate level of EMG amplitude seems to be controlled by (a) timing muscle activation at a latency timed from the expected instant of foot contact and (b) varying the rate at which EMG builds up.

This is a long awaited update of a much loved text, first produced in 1976. Bentley has retained a very successful format considering structures, actions and physiological roles of hormones across the vertebrate series. The evolution and origins of hormones and their receptors is discussed in the context of their contribution to major body systems and the ecology and evolutionary development of key vertebrate groups. In this way the integrative functions of hormones are examined in major body systems including nutrition, reproduction, osmoregulation, calcium homeostasis and in relation to animal colouration, moulting and development. Species considered range from representatives of lower chordates through the major fish groups to tetrapods including mammals.

The book, which is primarily a teaching aid aimed at a wide target audience, provides a short introductory course to the principles of molecular biology techniques.The drab appearance and cost of this manual belittle the wealth of easily accessed information presented in it. That, which appears to be yet another turgid introduction to the basics of molecular biology is, in fact, a technical handbook covering the whole spectrum of this complex subject in a simple way. The text provides a translation of the intricate subject of DNA technology and its terminology into everyday language. Each procedure is presented as blocks of information generously illustrated with self-explanatory figures which emphasize each point. The format allows incorporation of ideas and methodology into a teaching programme leading the student to understand not only the complexities of molecular biology but also its wider applications, for example its use in plant technology and medicine.