The aim of the present study was to examine the mechanisms of Ca2+ overload-induced contractile dysfunction in rat hearts independent of ischemia and acidosis. Experiments were performed on 30 excised cross-circulated rat heart preparations. After hearts were exposed to high Ca2+, there was a contractile failure associated with a parallel downward shift of the linear relation between myocardial O(2) consumption per beat and systolic pressure-volume area (index of a total mechanical energy per beat) in left ventricles from all seven hearts that underwent the protocol. This result suggested a decrease in O(2) consumption for total Ca2+ handling in excitation-contraction coupling. In the hearts that underwent the high Ca2+ protocol and had contractile failure, we found marked proteolysis of a cytoskeleton protein, alpha-fodrin, whereas other proteins were unaffected. A calpain inhibitor suppressed the contractile failure by high Ca2+, the decrease in O(2) consumption for total Ca2+ handling, and membrane alpha-fodrin degradation. We conclude that the exposure to high Ca2+ may induce contractile dysfunction possibly by suppressing total Ca2+ handling in excitation-contraction coupling and degradation of membrane alpha-fodrin via activation of calpain.

Recently, we have consistently observed curved endsystolic pressure-volume relations (ESPVRs) of the left ventricle (LV) in rat blood-perfused [1][2][3] and crystalloid-perfused whole heart preparations [4] and in situ ejecting rat hearts [5,6], like those of the puppy LV [7] and adult canine LV in supernormal contractility [8,9]. These studies suggest more generality of the curvilinear than linear ESPVR in different animal species. Despite this curvilinearity, we have obtained a linear myocardial oxygen consumption per Japanese Journal of Physiology, 49, 513-520, 1999 Key words: excitation-contraction coupling, oxygen consumption, E max (end-systolic pressure-volume ratio), systolic pressure-volume area (PVA). Abstract:We have already reported the linear oxygen consumption per beat (VO 2 )-systolic pressure-volume area (PVA) relation from the curved left ventricular (LV) end-systolic pressure-volume relation (ESPVR) in the cross-circulated rat heart. The VO 2 intercept (PVA-independent VO 2 ) is primarily composed of VO 2 for Ca 2ϩ handling in excitation-contraction (E-C) coupling and basal metabolism. The aim of the present study was to obtain the oxygen cost of LV contractility that indicates VO 2 for Ca 2ϩ handling in E-C coupling per unit LV contractility change in the rat heart. Oxygen cost of LV contractility is obtainable as a slope of a linear relation between PVA-independent VO 2 and LV contractility. We obtained a composite VO 2 -PVA relation line at a mid-range LV volume (mLVV) under gradually enhanced LV contractility by stepwise increased Ca 2ϩ infusion and thus the gradually increased PVA-independent VO 2 values. As a LV contractility index, we could not use E max (ESP-V ratio; ESP/ESV) for the linear ESPVR because of the curved ESPVR in the rat LV. A PVA at a mLVV (PVA mLVV ) has been proposed as a good index for assessing rat LV mechanoenergetics. Since the experimentally obtained PVA mLVV was not triangular due to the curved ESPVR, we propose an equivalent ESP-V ratio at a mLVV, (eESP/ ESV) mLVV , as a LV contractility index. This index was calculated as an ESP-V ratio of the specific virtual triangular PVA mLVV that is energetically equivalent to the real PVA mLVV . The present approach enabled us to obtain a linear relation between PVA-independent VO 2 and (eESP/ ESV) mLVV and the oxygen cost of LV contractility as the slope of this relation.

We conclude that (i) CVF prevents HAR, (ii) the addition of Spx + IS delays rejection, but (iii) the early deposition of antibody leads to progressive graft injury, resulting in (iv) delayed vascular rejection. Our findings indicate that the features of delayed xenograft rejection described in small animal models also occur in the pig-to-baboon model, and that rejection may occur in a complement-independent manner from the effects of antibody and/or host macrophages.

To gain insight into the pathogenesis of diabetic cardiomyopathy, we investigated cardiac function in terms of the coupling of left ventricular mechanical work and the energetics in Otsuka Long-Evans Tokushima Fatty rats, which are well known as a model of type 2 diabetes mellitus (DM). Neither left ventricular systolic function and mean coronary flow nor coronary flow reserve differed even in late DM rats. The amount of oxygen required for mechanical work and contraction was unaltered, although myosin isozyme was finally transformed from V(1) to V(3). The maximum pacing rate was decreased from 300 to 240 beats/min, and the left ventricular relaxation rate was significantly (P < 0.05) slower only in late DM rats, resulting in decreased oxygen consumption per minute for total Ca(2+) handling in excitation-contraction coupling mainly consumed by sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2) without significant changes in basal metabolism or in mitochondrial oxidative phosphorylation. The protein level of SERCA2 in membranes was significantly (P < 0.001) lower in severe DM rats. We conclude that the only lusitropic dysfunction due to the depressed expression of SERCA2 is related to generating diabetic cardiomyopathy even in the present type 2 diabetic rats.

a b s t r a c tWe examined the expression of the major H 2 S-producing enzymes, cystathionine-b-synthase (CBS) and cystathionine-c-lyase (CSE). CBS was ubiquitously distributed in the mouse pancreas, but CSE was found only in the exocrine. Freshly isolated islets expressed CBS, while CSE was faint. However, high glucose increased the CSE expression in the beta-cells. L-Cysteine or NaHS suppressed islet cell apoptosis with high glucose, and increased glutathione content in MIN6 beta-cells. Pretreatment with L-cysteine improved the secretory responsiveness following stimulation with glucose. The CSE inhibitor DL-propargylglycine antagonized these L-cysteine effects. We suggest H 2 S may function as an 'intrinsic brake' which protects beta-cells from glucotoxicity.

Patients who have DLT malposition after placing the patient in the lateral position had more DLT malposition during OLV and hypoxaemia during OLV.

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Mice carrying a null mutation in the glucokinase (GK) gene in pancreatic ␤-cells, but not in the liver, were generated by disrupting the ␤-cell-specific exon. Heterozygous mutant mice showed early-onset mild diabetes due to impaired insulin-secretory response to glucose. Homozygotes showed severe diabetes shortly after birth and died within a week. GK-deficient islets isolated from homozygotes showed defective insulin secretion in response to glucose, while they responded to other secretagogues: almost normally to arginine and to some extent to sulfonylureas. These data provide the first direct proof that GK serves as a glucose sensor molecule for insulin secretion and plays a pivotal role in glucose homeostasis. GK-deficient mice serve as an animal model of the insulin-secretory defect in human noninsulin-dependent diabetes mellitus.Glucokinase (GK), 1 mainly expressed in pancreatic ␤-cells and the liver, is thought to constitute a rate-limiting step in glucose metabolism in these tissues (1-4). Since insulin secretion parallels glucose metabolism and the high K m of GK (5-8 mM) ensures that it can change its enzymatic activity within the physiological range of glucose concentrations, GK has been proposed to act as a glucose sensor in the pancreatic ␤-cell (1, 5). Recently, mutations of the GK gene have been identified in patients with maturity-onset diabetes of the young, a subtype of early-onset non-insulin-dependent diabetes mellitus (NIDDM) (6 -8). However, since all the mutations in humans so far occur in the region of the gene that is common to pancreatic ␤-cells and hepatocytes (9), and are heterozygous, it may not have been possible to fully reveal physiological roles of pancreatic ␤-cell GK either in vivo or in vitro. To this end, mice carrying a null mutation in the GK gene in pancreatic ␤-cells, but not in the liver, were generated by homologous recombination. Heterozygous mutant mice showed early-onset mild diabetes resembling the phenotype for human maturity-onset diabetes of the young. Homozygotes showed severe diabetes shortly after birth and died within a week. GK-deficient islets showed defective insulin secretion in response to glucose, while they responded to other secretagogues: almost normally to arginine and to some extent to sulfonylureas. These data provide the first direct proof that GK serves as a glucose sensor molecule for insulin secretion and plays a pivotal role in glucose homeostasis. EXPERIMENTAL PROCEDURESCloning of the Mouse GK Gene, Construction of a Targeting Vector, and Homologous Recombinant Experiments-A DNA fragment including the pancreatic ␤-cell-specific exon 1␤ of the GK gene was cloned from a BALB/c mouse genomic library (Clontech). A BamHI site was introduced 30 base pairs 3Ј to the translation initiation codon of GK by the Kunkel method (10). A neomycin resistance gene (neo r ) with a pgk-1 promoter but without a poly(A) ϩ addition signal was substituted for the XbaI-BamHI fragment in the exon 1␤. A diphtheria toxin A fragment gene (DTA) with a MC1 promoter was lig...