Ultrasonic treatment stimulates multiple shoot regeneration and explant enlargement in recalcitrant squash cotyledon explants in vitro

Ultrasonic treatment (0.5–2 min) stimulated multiple shoot regeneration to high levels in vitro from recalcitrant cotyledon explants of commercial squash (Cucurbita pepo L.) cultivars Ma’yan and Bareqet, on Murashige and Skoog [Physiol Plant 15:473–497, 1962] (regeneration) medium augmented with 4.4 μM benzyladenine. At this stage, unsonicated control explants regenerated only a few very small shoots or bud-like structures. Ultrasound also stimulated massive explant growth. Ultrasound treatment resulted in further multiple shoot production (five times greater than control) after explant transfer to elongation medium (Murashige and Skoog [Physiol Plant 15:473–497, 1962] medium with 0.44 μM benzyladenine and 2.9 μM gibberellic acid). Longer ultrasonic treatments (5 or 10 min) promoted multiple shoot regeneration and explant growth accompanied by hyperhydration. Scanning electron microscope observations showed that 2 min ultrasound changed the joint area between epidermal cells and removed some of the surface from the cotyledon epidermal cells, without gross surface injury to the explants. Longer periods of ultrasound (5–10 min) caused further surface erosion. Rubbing the explant contact surface with chloroform or sandpaper emulated the effect of sonication on shoot regeneration and explant growth, demonstrating that ultrasound exerts its morphogenic influence by surface removal. Sonication of explants from other batches of squash seeds (of cultivars Ma'yan and True French), that regenerated without such treatment, reduced regeneration and caused hyperhydration. This is the first report of stimulation of in vitro regeneration by ultrasound treatment. Electronic Supplementary Material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Nutritional needs for exercise in the heat

Although hot conditions are not typically conducive to optimal sports performance, nutritional strategies play an important role in assisting an athlete to perform as well as possible in a hot environment. A key issue is the prevention of hypohydration during an exercise session. Fluid intake strategies should be undertaken in a cyclical sequence: hydrate well prior to the workout, drink as much as is comfortable and practical during the session, and rehydrate aggressively afterwards in preparation for future exercise bouts. There is some interest in hyperhydration strategies, such as hyperhydration with glycerol, to prepare the athlete for a situation where there is little opportunity for fluid intake to match large sweat losses. Recovery of significant fluid losses after exercise is assisted by the simultaneous replacement of electrolyte losses. Carbohydrate (CHO) requirements for exercise are increased in the heat, due to a shift in substrate utilization towards CHO oxidation. Daily food patterns should focus on replacing glycogen stores after exercise, and competition strategies should include activities to enhance CHO availability, such as CHO loading for endurance events, pre-event CHO intake, and intake of sports drinks in events lasting longer than 60 min. Although CHO ingestion may not enhance the performance of all events undertaken in hot weather, there are no disadvantages to the consumption of beverages containing 4-8% CHO and electrolytes. In fact, the palatability of these drinks may enhance the voluntary intake of fluid. Although there is some evidence of increased protein catabolism and cellular damage due to production of oxygen radicals during exercise in the heat, there is insufficient evidence to make specific dietary recommendations to account for these issues.     

Hydration effects on thermoregulation and performance in the heat

During exercise, sweat output often exceeds water intake, producing a water deficit or hypohydration. The water deficit lowers both intracellular and extracellular fluid volumes, and causes a hypotonic-hypovolemia of the blood. Aerobic exercise tasks are likely to be adversely effected by hypohydration (even in the absence of heat strain), with the potential affect being greater in hot environments. Hypohydration increases heat storage by reducing sweating rate and skin blood flow responses for a given core temperature. Hypertonicity and hypovolemia both contribute to reduced heat loss and increased heat storage. In addition, hypovolemia and the displacement of blood to the skin make it difficult to maintain central venous pressure and thus cardiac output to simultaneously support metabolism and thermoregulation. Hyperhydration provides no advantages over euhydration regarding thermoregulation and exercise performance in the heat.     

Effect of daily hyperhydration on fluid-electrolyte changes in endurance-trained volunteers during prolonged restriction of muscular activity

The aim of this investigation was to evaluate the effect of a daily intake of fluid and salt supplementation on fluid and electrolyte losses in endurance-trained volunteers during prolonged restriction of muscular activity (hypokinesia). The studies were performed on 30 long-distance runners aged 23–26 who had a peak oxygen uptake of 65.5 mL/kg/min and had taken 13.8 km/d on average prior to their participation in the study. The volunteers were divided into three groups: The volunteers in the first group were placed under normal ambulatory conditions (control subjects), the second group of volunteers subjected to hypokinesia alone (hypokinetic subjects), and the third group of volunteers was submitted to HK and consumed daily 0.1 g sodium chloride (NaCl)/kg body wt and 26 mL water/kg body wt (hyperhydrated subjects). The second and third group of volunteers were kept under an average of 2.7 km/d for 364 d. During the pre-experimental period of 60 d and during the experimental period of 364 d sodium, potassium, calcium, and magnesium in urine and plasma were determined. Blood was also assayed for osmolality, hemoglobin, hematocrit, plasma volume, plasma renin activity and plasma aldosterone. Mean arterial blood pressure was also determined. In the hyperhydrated volunteers plasma volume and arterial blood pressure increased, whereas plasma osmolality, plasma renin activity, plasma aldosterone, hematocrit, hemoglobin concentration, and urinary excretion and concentrations of electrolytes in plasma decreased. In the hypokinetic volunteers, plasma volume and arterial blood pressure decreased significantly, whereas hematocrit values, hemoglobin concenfration, plasma osmolality, plasma renin activity, plasma aldosterone, and electrolytes in urine and plasma increased significantly during the experimental period. It was concluded that chronic hyperhydration may be used in minimizing fluid and electrolyte losses in endurance-trained volunteers during prolonged restriction of muscular activity.