Carnitine for Body Composition in Hemodialysis Patients

Background: Authors and colleagues have continued clinical research for hemodialysis patients. Currently, a pilot study presents intervention of carnitine for changes of the body composition. Subjects and Methods: Subjects were six patients on hemodialysis with intervention of carnitine (group 1). Average data were 74.3 years, 65.4 kg, 22.6 in BMI. As levocarnitine, L-Cartin FF injection 1000 mg was administered three times a week for six months. Group 2 has six control patients for age-, sex-, body weight, BMI-matched (group 2). Body composition of muscle and fat tissues were measured by InBody 770 on 0 and 6 months. Results: In group 1, muscle volume and skeletal muscle showed increasing tendency without statistical significance. In contrast, there were significant decreases of body fat volume (22.3 kg vs 20.5 kg, 39.0% vs 35.8%) (p<0.05). No significant differences were found in hemoglobin, total protein, albumin and Cardio-Thoracic Ratio (CTR) of chest X-ray. Group 2 showed no significant changes. Discussion and Conclusion: Hemodialysis patients often have muscular reduction. Previous reports showed improved lean body mass by carnitine administration, which may support our result. These results from current pilot study would be expected to become useful reference data in the pathophysiological investigation in patients on hemodialysis

Investigation of Nerve Conduction in Patients with Diabetes and/or Hemodialysis

Diabetic peripheral neuropathy (DPN) has been clinically important, and nerve conduction studies (NCS) have been performed with rather complexity and high cost. By advances in technology, simple and useful DPN-Check device was developed obtaining NCS data as sural nerve conduction velocity (SNCV) and sural nerve action potential (SNAP). We enrolled 52 subjects classified into 4 groups according to the presence of hemodialysis (HD) and diabetes mellitus (DM) as follows: HD (+), DM (+) in group 1, HD (+), DM (-) in group 2, HD (-), DM (+) in group 3 and healthy controls in group 4. Average age was similar from 68 to 74 years in 4 groups. Median value of SNCV was 31, 48, 49, 54 m/sec, and median value of SNAP was 3, 9, 6, 22 μV, respectively, in 4 groups. These results might suggest some relationship between impaired states of HD and DM, and would become fundamental data for pathophysiological investigation of peripheral neuropathy of HD and/or DM in the future

Investigation of Nerve Conduction in Patients with Diabetes and/or Hemodialysis

Diabetic peripheral neuropathy (DPN) has been clinically important, and nerve conduction studies (NCS) have been performed with rather complexity and high cost. By advances in technology, simple and useful DPN-Check device was developed obtaining NCS data as sural nerve conduction velocity (SNCV) and sural nerve action potential (SNAP). We enrolled 52 subjects classified into 4 groups according to the presence of hemodialysis (HD) and diabetes mellitus (DM) as follows: HD (+), DM (+) in group 1, HD (+), DM (-) in group 2, HD (-), DM (+) in group 3 and healthy controls in group 4. Average age was similar from 68 to 74 years in 4 groups. Median value of SNCV was 31, 48, 49, 54 m/sec, and median value of SNAP was 3, 9, 6, 22 μV, respectively, in 4 groups. These results might suggest some relationship between impaired states of HD and DM, and would become fundamental data for pathophysiological investigation of peripheral neuropathy of HD and/or DM in the future

Influence of Diabetes and Hemodialysis Against Nerve Conduction Studies

Background: Diabetic peripheral neuropathy (DPN) has been prevalent and discussed, and nerve conduction studies (NCS) has been continued. We have checked NCS using recently introduced useful DPN-Check device. Subjects and Methods: The subjects were 66 patients (pts) classified into 4 groups according to existence of diabetes mellitus (DM) and hemodialysis (HD); Group1: DM (+), HD (+) in 15 pts, group 2: DM (-), HD (+) in 15 pts, group 3: DM (+), HD (-) in 20 pts, group 4: 16 healthy controls. Methods included measurements of sural nerve conduction velocity (SNCV) and sural nerve action potential (SNAP) using HDN-1000. Results: Average age in each group was 64.4 years to 72.6 years. SNCV value of 4 group in average was 37.1 m/sec, 46.3 m/sec, 49.3 m/sec, 53.2 m/sec, respectively, and value of group 1 was significantly lower than those of group 2,3,4 (p<0.01). Similarly, average SNAP was 4.1 μV, 8.7 μV, 8.0 μV, 21.6 μV, respectively, and group 1,2,3 were significantly lower than group 4 (p<0.01). There was significant correlation between SNCV and SNAP in all subjects (p<0.01). Significant correlations were shown between DM duration and SNCV, and DM duration and SNAP (p<0.01). Discussion and Conclusion: SNCV and SNAP were measured successfully and easily by HDN-1000, indicating clinical availability. Obtained data suggested that 1) SNCV is not significantly decreased due to only uremic neuropathy, 2) SNCV is significantly decreased in patients with both HD and DM, 3) SNAP is significantly decreased in patents with DM for years and 4) SNAP would be remarkably decreased when HD is in addition to DM. These results would become the basal data of future NCS for DM and HD

Vascular Smooth Muscle Cells Stimulate Platelets and Facilitate Thrombus Formation through Platelet CLEC-2: Implications in Atherothrombosis

<div><p>The platelet receptor CLEC-2 is involved in thrombosis/hemostasis, but its ligand, podoplanin, is expressed only in advanced atherosclerotic lesions. We investigated CLEC-2 ligands in vessel walls. Recombinant CLEC-2 bound to early atherosclerotic lesions and normal arterial walls, co-localizing with vascular smooth muscle cells (VSMCs). Flow cytometry and immunocytochemistry showed that recombinant CLEC-2, but not an anti-podoplanin antibody, bound to VSMCs, suggesting that CLEC-2 ligands other than podoplanin are present in VSMCs. VSMCs stimulated platelet granule release and supported thrombus formation under flow, dependent on CLEC-2. The time to occlusion in a FeCl₃-induced animal thrombosis model was significantly prolonged in the absence of CLEC-2. Because the internal elastic lamina was lacerated in our FeCl₃-induced model, we assume that the interaction between CLEC-2 and its ligands in VSMCs induces thrombus formation. Protein arrays and Biacore analysis were used to identify S100A13 as a CLEC-2 ligand in VSMCs. However, S100A13 is not responsible for the above-described VSMC-induced platelet activation, because S100A13 is not expressed on the surface of normal VSMCs. S100A13 was released upon oxidative stress and expressed in the luminal area of atherosclerotic lesions. Suspended S100A13 did not activate platelets, but immobilized S100A13 significantly increased thrombus formation on collagen-coated surfaces. Taken together, we proposed that VSMCs stimulate platelets through CLEC-2, possibly leading to thrombus formation after plaque erosion and stent implantation, where VSMCs are exposed to blood flow. Furthermore, we identified S100A13 as one of the ligands on VSMCs.</p></div

S100A13 is not expressed on the surface of coronary artery smooth muscle cells (CASMCs), but oxidative stress induced surface expression of S100A13.

<p>A) Binding of recombinant S100A13 on the surface of CASMCs were examined by flow cytometory. CASMCs were preincubated with vehicle only (1st panel) or recombinant S100A13 in the presence of vehicle (2^nd panel), 0.1 mM CaCl₂ (3^rd panel), 1 mM EDTA (4^th panel), or 0.1 mM CaCl₂ + 1 mM EDTA (5^th panel). After excess of the recombinant protein was removed by centrifugation, cells were incubated with control mouse IgG (filled) or anti-S100A13 antibody (line), followed by Alexa Flour 488-conjugated anti-mouse IgG. B) Surface expression of endogenous S100A13 was analyzed by flow cytometry. CASMCs pretreated with indicated concentrations of H₂O₂ were incubated with control mouse IgG (filled) or anti-S100A13 antibody (line), followed by Alexa Flour 488-conjugated anti-mouse IgG.</p

Coronary artery smooth muscle cells (CASMCs) stimulated release of α and dense granule contents through CLEC-2.

<p>Washed platelets obtained from the CLEC-2-deficient chimeras were incubated with buffer, CASMCs, CRP, rhodocytin, or lysis buffer for 10 min. The platelets were centrifuged. The amount of secreted 5-hydroxytryptamine (5-HT) (A) or platelet-derived growth factor (PDGF) (B) in the supernatants was measured by ELISA. Platelet lysates were used to measure the total amount of 5-HT or PDGF stored in platelets. The results were expressed as the percentage of secreted 5-HT or PDGF relative to the total amount stored in platelets ± SE (A. n = 9 from three independent experiments, B. n = 12 from four independent experiments). Three asterisks denote p < 0.005.</p

S100A13 potentiated thrombus formation on collagen-coated surfaces under flow.

<p>(A) Anti-coagulated human whole blood was perfused into cell microscopy chambers coated with collagen or collagen plus S100A13 for 10 min at a shear rate of 1500 s⁻¹. Adherent platelets were visualized using a fluorescence video microscope. Movie data were converted into sequential photo images. (B) Thrombus volume was quantified. After 5 min of perfusion, adherent platelets were visualized by confocal laser microscopy, and the z-stack data were analyzed. The thrombus volume was expressed as the cIFI per image (404374 μm²). The graph illustrates the percentage of the surfaces coated with collagen only cIFI ± SE (n = 6, from two independent experiments).</p

Time to thrombotic occlusion of vessels stimulated by FeCl₃, but not by green light and dye, was significantly delayed in the CLEC-2-deficient chimeras.

<p>The femoral artery of anesthetized wild type (WT) chimeras or CLEC-2-deficient chimeras was exposed by blunt dissection. A) Time to vessel occlusion in the FeCl₃ model: a 1strip of filter paper saturated with 10% FeCl₃ was applied to the adventitial surface of the exposed femoral artery. B) Time to occlusion in the photochemically induced thrombosis (PIT) model. After 5 min of irradiation to the exposed femoral artery, rose Bengal was infused. The time to thrombotic vessel occlusion of the artery was measured by monitoring femoral blood flow using a Doppler blood flow velocimeter. Each symbol represents one individual. The mean time to occlusion ± SE is indicated in the graphs. Two asterisks denote p < 0.01.</p