A microplate technique to simultaneously assay calcium accumulation in endoplasmic reticulum and SERCA release of inorganic phosphate

Traditional analyses of calcium homeostasis have separately quantified either calcium accumulation or release mechanisms. To define the system as a whole, however, requires multiple experimental techniques to examine both accumulation and release. Here we describe a technique that couples the simultaneous quantification of radio-labeled calcium accumulation in endoplasmic reticulum (ER) microsomes with the release of inorganic phosphate (Pi) by the hydrolytic activity of sarco-endoplasmic reticulum calcium ATPase (SERCA) all in the convenience of a 96-well format

Measurement of resting energy expenditure in infants

Objective: The method for measurement of basal metabolic rate (BMR) using indirect calorimetry in adults is well established but is impractical in infants. Methods: In this prospective study energy expenditure was measured using indirect calorimetry in 14 infants when sleeping and when lying quietly awake. Results: Sleeping metabolic rate (SMR) was lower than energy expenditure (EE) measured in the same infants in a quiet resting state (mean difference [SD]: 297 [162] kJ/d; P < 0.005; 55 [33.4] kJ/kg per day; P < 0.005). The correlation within individuals suggests that these differences are related to the level of arousal. Awake EE, but not SMR, was significantly greater than estimated BMR using the FAO/WHO/UNU predictive equation. Conclusions: In infants, the level of arousal during measurement of EE can significantly impact on the interpretation of EE results. A standardized method for the measurement of EE in infants using indirect calorimetry is proposed

A novel vasopressin-induced transcript promotes MAP kinase activation and ENaC downregulation

In the principal cell of the renal collecting duct, vasopressin regulates the expression of a gene network responsible for sodium and water reabsorption through the regulation of the water channel and the epithelial sodium channel (ENaC). We have recently identified a novel vasopressin-induced transcript (VIT32) that encodes for a 142 amino acid vasopressin-induced protein (VIP32), which has no homology with any protein of known function. The Xenopus oocyte expression system revealed two functions: (i) when injected alone, VIT32 cRNA rapidly induces oocyte meiotic maturation through the activation of the maturation promoting factor, the amphibian homolog of the universal M phase trigger Cdc2/cyclin; and (ii) when co-injected with the ENaC, VIT32 cRNA selectively downregulates channel activity, but not channel cell surface expression. In the kidney principal cell, VIP32 may be involved in the downregulation of transepithelial sodium transport observed within a few hours after vasopressin treatment. VIP32 belongs to a novel gene family ubiquitously expressed in oocyte and somatic cells that may be involved in G to M transition and cell cycling