Clinical Outcomes of Living Liver Transplantation According to the Presence of Sarcopenia as Defined by Skeletal Muscle Mass, Hand Grip, and Gait Speed.

BACKGROUND: Sarcopenia is an independent predictor of death after living-donor liver transplantation (LDLT). However, the ability of the Asian Working Group for Sarcopenia criteria for sarcopenia (defined as reduced skeletal muscle mass plus low muscle strength) to predict surgical outcomes in patients who have undergone LDLT has not been determined. METHODS: This study prospectively enrolled 366 patients who underwent LDLT at Kyushu University Hospital. Skeletal muscle area (determined by computed tomography), hand-grip strength, and gait speed were measured in 102 patients before LDLT. We investigated the relationship between sarcopenia and surgical outcomes after LDLT performed in three time periods. RESULTS: The number of patients with lower skeletal muscle area has increased to 52.9% in recent years. The incidence of sarcopenia according to the Asian Working Group for Sarcopenia criteria was 23.5% (24/102). Patients with sarcopenia (defined by skeletal muscle area and functional parameters) had significantly lower skeletal muscle area and weaker hand-grip strength than did those without sarcopenia. Compared with non-sarcopenic patients, patients with sarcopenia also had significantly worse liver function, greater estimated blood loss, greater incidence of postoperative complications of Clavien-Dindo grade IV or greater (including amount of ascites on postoperative day 14, total bilirubin on postoperative day 14, and postoperative sepsis), and longer postoperative hospital stay. Multiple logistic regression analysis revealed sarcopenia as a significant predictor of 6-month mortality. CONCLUSIONS: The combination of skeletal muscle mass and function can predict surgical outcomes in LDLT patients.

Caveolar disruption causes contraction of rat femoral arteries via reduced basal NO release and subsequent closure of BKCa channels.

Background and Purpose. Caveolae act as signalling hubs in endothelial and smooth muscle cells. Caveolar disruption by the membrane cholesterol depleting agent methyl-ß-cyclodextrin (M-ß-CD) has various functional effects on arteries including (i) impairment of endothelium-dependent relaxation, and (ii) alteration of smooth muscle cell (SMC) contraction independently of the endothelium. The aim of this study was to explore the effects of M-ß-CD on rat femoral arteries. Methods. Isometric force was measured in rat femoral arteries stimulated to contract with a solution containing 20 mM K(+) and 200 nM Bay K 8644 (20 K/Bay K) or with one containing 80 mM K(+)(80 K). Results. Incubation of arteries with M-ß-CD (5 mM, 60 min) increased force in response to 20 K/Bay K but not that induced by 80 K. Application of cholesterol saturated M-ß-CD (Ch-MCD, 5 mM, 50 min) reversed the effects of M-ß-CD. After mechanical removal of endothelial cells M-ß-CD caused only a small enhancement of contractions to 20 K/Bay K. This result suggests M-ß-CD acts via altering release of an endothelial-derived vasodilator or vasoconstrictor. When nitric oxide synthase was blocked by pre-incubation of arteries with L-NAME (250 µM) the contraction of arteries to 20 K/Bay K was enhanced, and this effect was abolished by pre-treatment with M-ß-CD. This suggests M-ß-CD is inhibiting endothelial NO release. Inhibition of large conductance voltage- and Ca(2+)-activated (BKCa) channels with 2 mM TEA(+) or 100 nM Iberiotoxin (IbTX) enhanced 20 K/Bay K contractions. L-NAME attenuated the contractile effect of IbTX, as did endothelial removal. Conclusions. Our results suggest caveolar disruption results in decreased release of endothelial-derived nitric oxide in rat femoral artery, resulting in a reduced contribution of BKCa channels to the smooth muscle cell membrane potential, causing depolarisation and contraction.

The incidence and mechanism of sunitinib-induced thyroid atrophy in patients with metastatic renal cell carcinoma.

BACKGROUND: To elucidate the incidence and mechanisms of sunitinib-induced thyroid atrophy, we investigated serial volumetric and functional changes, and evaluated histological changes of the thyroid gland in metastatic renal cell carcinoma patients who received sunitinib. METHODS: Thyroid volume (by computed tomography volumetry) and thyroid function were measured at baseline, during the treatment, and at post-treatment periods. Histological evaluation of the thyroid gland was performed in four autopsied patients. RESULTS: The median reduction rate in thyroid volume at last evaluation during sunitinib treatment was 30% in all 17 patients. The incidence of hypothyroidism during sunitinib treatment was significantly higher in the high reduction rate group (n=8; more than 50% reduction in volume) than in the low reduction rate group (n=9; less than 50% reduction in volume). Half of the patients in the high reduction rate group exhibited a transient thyroid-stimulating hormone suppression, suggesting thyrotoxicosis during sunitinib treatment. Histological evaluation demonstrated atrophy of thyroid follicles and degeneration of follicular epithelial cells without critical diminution of vascular volume in the thyroid gland. CONCLUSION: Thyroid atrophy is frequently observed following sunitinib treatment and may be brought about by sunitinib-induced thyrotoxicosis or the direct effects of sunitinib that lead to degeneration of thyroid follicular cells.

Caveolin-1 and caveolin-3 regulate Ca2+ homeostasis of single smooth muscle cells from rat cerebral resistance arteries.

The role of caveolins, signature proteins of caveolae, in arterial Ca(2+) regulation is unknown. We investigated modulation of Ca(2+) homeostasis by caveolin-1 and caveolin-3 using smooth muscle cells from rat cerebral resistance arteries. Membrane current and Ca(2+) transients were simultaneously measured with voltage-clamped single cells. Membrane depolarization triggered Ca(2+) current and increased intracellular Ca(2+) concentration ([Ca(2+)](i)). After repolarization, elevated [Ca(2+)](i) returned to the resting level. Ca(2+) removal rate was determined from the declining phase of the Ca(2+) transient. Application of caveolin-1 antibody or caveolin-1 scaffolding domain peptide, corresponding to amino acid residues 82-101 of caveolin-1, significantly slowed Ca(2+) removal rate at a measured [Ca(2+)](i) of 250 nM, with little effect at a measured [Ca(2+)](i) of 600 nM. Application of caveolin-3 antibody or caveolin-3 scaffolding domain peptide, corresponding to amino acid residues 55-74 of caveolin-3, also significantly slowed Ca(2+) removal rate at a measured [Ca(2+)](i) of 250 nM, with little effect at a measured [Ca(2+)](i) of 600 nM. Likewise, application of calmodulin inhibitory peptide, autocamtide-2-related inhibitory peptide, and cyclosporine A, inhibitors for calmodulin, Ca(2+)/calmodulin-dependent protein kinase II, and calcineurin, also significantly inhibited Ca(2+) removal rate at a measured [Ca(2+)](i) of 250 nM but not at 600 nM. Application of cyclopiazonic acid, a sarcoplasmic reticulum Ca(2+) ATPase inhibitor, also significantly inhibited Ca(2+) removal rate at a measured [Ca(2+)](i) of 250 nM but not at 600 nM. Our results suggest that caveolin-1 and caveolin-3 are important in Ca(2+) removal of resistance artery smooth muscle cells.

Effects of hypoxia, anoxia, and metabolic inhibitors on KATP channels in rat femoral artery myocytes.

Vascular ATP-sensitive potassium (KATP) channels have an important role in hypoxic vasodilation. Because KATP channel activity depends on intracellular nucleotide concentration, one hypothesis is that hypoxia activates channels by reducing cellular ATP production. However, this has not been rigorously tested. In this study we measured KATP current in response to hypoxia and modulators of cellular metabolism in single smooth muscle cells from the rat femoral artery by using the whole cell patch-clamp technique. KATP current was not activated by exposure of cells to hypoxic solutions (Po2 approximately 35 mmHg). In contrast, voltage-dependent calcium current and the depolarization-induced rise in intracellular calcium concentration ([Ca2+]i) was inhibited by hypoxia. Blocking mitochondrial ATP production by using the ATP synthase inhibitor oligomycin B (3 microM) did not activate current. Blocking glycolytic ATP production by using 2-deoxy-D-glucose (5 mM) also did not activate current. The protonophore carbonyl cyanide m-chlorophenylhydrazone (1 microM) depolarized the mitochondrial membrane potential and activated KATP current. This activation was reversed by oligomycin B, suggesting it occurred as a consequence of mitochondrial ATP consumption by ATP synthase working in reverse mode. Finally, anoxia induced by dithionite (0.5 mM) also depolarized the mitochondrial membrane potential and activated KATP current. Our data show that: 1) anoxia but not hypoxia activates KATP current in femoral artery myocytes; and 2) inhibition of cellular energy production is insufficient to activate KATP current and that energy consumption is required for current activation. These results suggest that vascular KATP channels are not activated during hypoxia via changes in cell metabolism. Furthermore, part of the relaxant effect of hypoxia on rat femoral artery may be mediated by changes in [Ca2+]i through modulation of calcium channel activity.

Molecular analysis of the subtype-selective inhibition of cloned KATP channels by PNU-37883A.

1. In this study, we have used Kir6.1/Kir6.2 chimeric proteins and current recordings to investigate the molecular basis of PNU-37883A inhibition of cloned K(ATP) channels. 2. Rat Kir6.1, Kir6.2 and Kir6.1/Kir6.2 chimeras were co-expressed with either SUR2B or SUR1, following RNA injection into Xenopus oocytes, and fractional inhibition of K(ATP) currents by 10 microm PNU-37883A reported. 3. Channels containing Kir6.1/SUR2B were more sensitive to inhibition by PNU-37883A than those containing Kir6.2/SUR2B (mean fractional inhibition: 0.70, cf. 0.07). 4. On expression with SUR2B, a chimeric channel with the Kir6.1 pore and the Kir6.2 amino- and carboxy-terminal domains was PNU-37883A insensitive (0.06). A chimera with the Kir6.1 carboxy-terminus and Kir6.2 amino-terminus and pore was inhibited (0.48). These results, and those obtained with other chimeras, suggest that the C-terminus is an important determinant of PNU-37883A inhibition of Kir6.1. Similar results were seen when constructs were co-expressed with SUR1. Further chimeric constructs localised PNU-37883A sensitivity to an 81 amino-acid residue section in the Kir6.1 carboxy-terminus. 5. Our data show that structural differences between Kir6.1 and Kir6.2 are important in determining sensitivity to PNU-37883A. This compound may prove useful in probing the structural and functional differences between the two channel subtypes.

Ca2+-induced Ca2+ release in cardiac and smooth muscle cells.

Ca(2+) influx across plasma membranes may trigger Ca(2+) release by activating ryanodine-sensitive receptors in the sarcoplasmic reticulum. This process is called Ca(2+)-induced Ca(2+) release, and may be important in regulating cytosolic Ca(2+) concentration ([Ca(2+)](i)). In cardiac cells, the initial [Ca(2+)](i) increase, caused by L-type Ca(2+) current, is profoundly amplified with Ca(2+) release. The synchronized opening of several ryanodine-sensitive Ca(2+)-releasing channels was detected as discreet and highly localized Ca(2+) elevation, and termed as a 'Ca(2+) spark'. A Ca(2+) spark is under local control of an L-type Ca(2+) channel, and therefore a Ca(2+) spark does not normally trigger subsequent Ca(2+) sparks in the neighbouring area. In smooth muscle cells, the importance of Ca(2+)-induced Ca(2+) release in elevating [Ca(2+)](i) appears to differ among preparations and species. Significant elevation in [Ca(2+)](i) during depolarization was attributed to Ca(2+) release in some smooth muscle cells, but not in others. Ca(2+) sparks are also identified in smooth muscle cells, and may play a role as functional elementary events for Ca(2+)-induced Ca(2+) release. At rest, Ca(2+) sparks may be also important in regulating smooth muscle membrane potential. Ca(2+) sparks occurring at rest do not raise global [Ca(2+)](i), but trigger spontaneous transient outward currents (STOCs) or spontaneous transient inward currents (STICs), the former producing hyperpolarization; the latter, depolarization. Thus there may be multiple facets for Ca(2+)-induced Ca(2+) release in regulating the contractile status of smooth muscle cells.

Rapid MR imaging detection of renal cysts: age-based standards.

PURPOSE: To establish age-based standards for renal cysts depicted at magnetic resonance (MR) imaging and to compare these standards with existing standards for ultrasonography (US). MATERIALS AND METHODS: Three radiologists reviewed subsecond T2-weighted single-shot fast spin-echo kidney MR imaging findings in 528 patients (248 men, 280 women) selected from consecutive abdominal MR studies without regard to clinical indication. Age, sex, and number and diameter of cysts were noted. Results were analyzed with nonparametric tests and were compared with published US results. RESULTS: Men (mean, 2.0; 95% CI: 1.5, 2.5) had more renal cysts than women (mean, 1.2; 95% CI: 0.9, 1.5) (P < .001). Number and diameter of cysts increased with age (P < .001). Of 528 patients, 330 (62.5%) had at least one renal cyst, and 315 (59.7%) had cysts of 10 mm or less. MR imaging findings were comparable to published US criteria for type 1 autosomal dominant polycystic kidney disease (ADPKD) if only cysts larger than 1 cm were considered: Only one subject in the group of 18-29-year-old subjects had at least two renal cysts, and five of 493 subjects aged 30-59 years had at least two cysts in each kidney. CONCLUSION: Compared with reported US results, MR imaging depicted an increased number of simple renal cysts in healthy individuals because of its increased sensitivity for cysts smaller than 1 cm. If only simple renal cysts larger than 1 cm are considered, US criteria for type 1 ADPKD can be applied to MR imaging.

Elbow synovial fold syndrome: MR imaging findings.

OBJECTIVE: We describe the anatomy and MR imaging appearance of elbow plicae. MATERIALS AND METHODS: First, five cadavers were evaluated by sectioning and using MR arthrography for evidence of normal or prominent synovial folds to determine the potential origin of elbow plicae. Next, 164 consecutive MR images were evaluated to determine the frequency of the plicae in a clinical population. Last, we retrospectively studied a selected group of eight patients who underwent preoperative MR imaging and in whom enlarged synovial folds were confirmed at surgery. RESULTS: In the cadavers, the synovial fold appeared to originate from the synovium adjacent to a posterior fat pad. In the clinical population, half the patients showed a synovial fold at the same location; however, most folds were less than or equal to 2 mm in thickness. The eight patients presented clinically with symptoms mimicking an intraarticular body. The synovial fold in symptomatic patients was seen posteriorly just above the olecranon and averaged 3 mm in thickness. CONCLUSION: A synovial fold extending from the posterior fat pad in the elbow is a frequent finding on MR imaging. In a subgroup of patients, plicae, when thickened, may present clinically as a locking elbow. However, overlap exists between the thicknesses of symptomatic and asymptomatic plicae.

Utilization of "used" vials: cost-effective technique for MR arthrography.

Because full vials of commercially available MR arthrographic contrast are expensive, we hypothesized that the small residual contrast in a "used" vial would be adequate for MR arthrography. After sterility testing and quantity analysis of the residual contrast in 28 vials, this method was successfully used in 10 patients. J. Magn. Reson. Imaging 2000;12:953-955.

Mechanisms that regulate [Ca2+]i following depolarization in rat systemic arterial smooth muscle cells.

1. We have used the patch-clamp technique in combination with fluorimetric recording to study the mechanisms that regulate intracellular Ca2+, [Ca2+]i, following depolarization in cells isolated from the rat femoral artery. 2. Depolarization to 0 mV from a holding potential of -70 mV increased [Ca2+]i. Little Ca2+ release from sarcoplasmic reticulum, SR, was detected during depolarization since application of 30 microM ryanodine, a Ca2+-release inhibitor, had no significant effect on total Ca2+ buffering power. 3. Upon repolarization to -70 mV, 7 out of 13 cells showed three phases of Ca2+ removal; an initial rapid first phase, a slow second phase, and a faster third phase. Six cells, in which Ca2+ recovered quickly, lacked the third phase. The third phase was also absent in cells treated with a SR Ca2+-pump inhibitor, cyclopiazonic acid. 4. The peak first-phase Ca2+ removal rate observed upon repolarization to -70 mV was significantly reduced in cells treated with a mitochondrial Ca2+ uptake inhibitor, carbonyl cyanide m-chlorophenylhydrazone. However, an ATP-synthase inhibitor, oligomycin B, had no significant effect. 5. The Ca2+ removal rate was little affected by clamping the cell at +120 mV rather than -70 mV, suggesting that Ca2+ removal processes are largely voltage independent. Also, little inward current was associated with Ca2+ clearance, indicating that Ca2+ removal does not involve an electrogenic process. 6. Our results suggest that Ca2+-induced Ca2+ release contributes little to the elevation of Ca2+ in these cells. The SR Ca2+ pump may contribute to Ca2+ removal over a low [Ca2+]i range in cells where [Ca2+]i remains high for long enough, while mitochondrial Ca2+ uptake may be important when [Ca2+]i is high.

Pigmented hidrocystoma of the eccrine secretory coil in the vulva: clinicopathologic, immunohistochemical and ultrastructural studies.

A case of pigmented hidrocystoma of eccrine secretory coil is presented. A 47-year-old woman had developed a bluish black small nodule in the anterior portion of the labium minor a few years before entry. Microscopically, the cyst was lined by eosinophilic columnar epithelium with abundant brownish granules. There was a vague suggestion of decapitation secretion focally in the epithelial layer of cuboidal cells. This layer expressed distinct reactivity against CA19-9 with no reactivity for human milk fat globule-1 (HMFG-1). These features demonstrated that the cyst was not of apocrine nature but of eccrine derivation. In addition, positive immunoreaction for cytokeratin (CK)7, CK8 and CK19 defined the cyst as originating from the secretory coil of the sweat gland. Ultrastructurally, melanosomes in various stages were identified in most of the epithelial cells. These findings suggest that the present case was a hidrocystoma of eccrine secretory coil with abnormal melanin accumulation.

Ca2+ removal mechanisms in rat cerebral resistance size arteries.

Tissue blood flow and blood pressure are each regulated by the contractile behavior of resistance artery smooth muscle. Vascular diseases such as hypertension have also been attributed to changes in vascular smooth muscle function as a consequence of altered Ca2+ removal. In the present study of Ca2+ removal mechanisms, in dissociated single cells from resistance arteries using fura-2 microfluorimetry and voltage clamp, Ca2+ uptake by the sarcoplasmic reticulum and extrusion by the Ca2+ pump in the cell membrane were demonstrably important in regulating Ca2+. In contrast, the Na+-Ca2+ exchanger played no detectable role in clearing Ca2+. Thus a voltage pulse to 0 mV, from a holding potential of -70 mV, triggered a Ca2+ influx and increased intracellular Ca2+ concentration ([Ca2+]i). On repolarization, [Ca2+]i returned to the resting level. The decline in [Ca2+]i consisted of three phases. Ca2+ removal was fast immediately after repolarization (first phase), then plateaued (second phase), and finally accelerated just before [Ca2+]i returned to resting levels (third phase). Thapsigargin or ryanodine, which each inhibit Ca2+ uptake into stores, did not affect the first but significantly inhibited the third phase. On the other hand, Na+ replacement with choline+ did not affect either the phasic features of Ca2+ removal or the absolute rate of its decline. Ca2+ removal was voltage-independent; holding the membrane potential at 120 mV, rather than at -70 mV, after the voltage pulse to 0 mV, did not attenuate Ca2+ removal rate. These results suggest that Ca2+ pumps in the sarcoplasmic reticulum and the plasma membrane, but not the Na+-Ca2+ exchanger, are important in Ca2+ removal in cerebral resistance artery cells.

Reactivity of antibody YU-311 in formalin-fixed, paraffin-embedded specimens of normal organs, non-hematopoietic tumors, and mast cell tumors.

YU-311 is a monoclonal antibody that reacts with a human leukemia cell line resistant for cytosine arabinoside and that identifies a 92 kDa membrane protein. The reactivity of YU-311 in normal organs, various non-hematopoietic tumors and in mast cell tumors in formalin-fixed, paraffin-embedded specimens was examined using immunohistochemical methods. In normal organs, YU-311 reacted with fundic glands of the stomach, the intercalated duct of the pancreas, the distal portion and the loop of Henle of renal tubules and tissue mast cells. Benign neoplasms of various organs showed no immunoreaction with YU-311, except for mast cell tumors. Some types of malignant neoplasms were occasionally positive against YU-311, suggesting neoplasms arising from or differentiating along normal YU-311-positive counterparts. Some other types of malignancies were rarely positive for YU-311, although their normal counterparts showed no immunoreactivity with YU-311. None of the non-epithelial tumors reacted with YU-311, except for one case of malignant melanoma. In contrast, normal tissue mast cells and their related tumors, such as urticaria pigmentosa or solitary mastocytoma, were constantly positive for YU-311. None of the non-hematopoietic human tumor cell lines examined in the present study was reactive with YU-311. These findings indicate that YU-311 is a good marker of some types of tumors and mast cell tumors and that an aberrant expression of YU-311 rarely occurs.

Small cell carcinoma arising from the outer urethral orifice: a case report examined by histologic, ultrastructural and immunohistochemical methods.

A case of small cell carcinoma arising in the outer urethral orifice is presented. The resected tumor showed a proliferation of small round or fusiform neoplastic cells in the submucosa. Tumor cells were arranged in sheets or a trabecular manner and possessed markedly hyperchromatic nuclei with a high N:C ratio, closely resembling small cell carcinoma of the lung. Characteristically, pagetoid intraepithelial spreading could be identified. However, there was no evidence of in situ transitional cell carcinoma and adeno- or squamous cell carcinoma components anywhere. Ultrastructurally, each tumor cell contained only a few membrane-bound cored granules measuring 60-100 nm, which were compatible with neurosecretory granules, and desmosome-like intercellular attachments, but lacked aggregated microfilaments. By immunohistochemical examination, tumor cells were positive for epithelial markers, such as cytokeratin and epithelial membrane antigen, and neuron specific enolase, but negative for any other neuro-endocrine markers. Extensive systemic examination failed to show the primary site to be other than the outer urethral orifice. These findings indicate that the current tumor is a small cell carcinoma with neuro-endocrine differentiation arising from the outer urethral orifice.

Pigmented renal cell carcinoma: accumulation of abnormal lysosomal granules.

AIMS: Five cases of renal cell carcinoma (RCC) with abnormal pigmentation have been examined by histochemical, immunohistochemical and ultrastructural methods. METHODS AND RESULTS: Compared with conventional RCCs, there was no difference in histological findings of each case, except for the presence of pigmented cells. In three cases, tumour cells possessing various sized brown granules with neuromelanin-like features were scattered throughout the tumour. The granules observed in one of the others were angulated lysosomes and in another tumour cells in the pigmented areas possessed the granules closely resembling those of granular cell tumour. However, melanosome or neurosecretory granules could not be detected in any of the cases examined. In three cases, some of these abnormal granules showed a weak acid phosphatase activity. On immunohistochemical examination, tumour cells showed a positive immunoreaction for epithelial markers and lacked any antigens suggesting neuroectodermal or neuroendocrine differentiation. The granules in three cases were faintly positive for lysozyme and KP-1. CONCLUSIONS: These findings indicate that abnormal pigmentation of RCCs examined in this study is attributed to accumulation of abnormal lysosomal granules in the neoplastic cells.

Resistance to apoptosis induced by serum depletion and all-trans retinoic acid in drug-resistant leukemic cell lines.

The relation between resistance to anticancer drugs and resistance to apoptosis has been investigated in the human leukemic cell line(KY-821) and its drug-resistant sublines. Under serum depletion conditions, drug-resistant cell lines showed apoptotic resistance when compared with the parental cell line. Drug resistant cell lines also showed resistance to apoptosis when treated with all-trans retinoic acid. DNA fragmentation was low in drug resistant cell lines under both stimulations. Flowcytometry analysis did not show any alterations of the Fas antigen, p53, bcl-2 and c-myc protein expression toward inhibition of apoptotic response in drug-resistant sublines. These results indicate that drug-resistant leukemic cells still show resistance to apoptosis-inducing stimulation such as poor nutrition and differentiation-inducing agents.