An in vitro evaluation of the glycation potential of a natural disaccharide,trehalose

Background. Glucose, an osmotic agent generally used in continuous ambulatory peritoneal dialysis (CAPD) dialysate, has a critical characteristic of forming advanced glycation endproducts (AGEs). We undertook this study to investigate whether a possible osmotic agent, trehalose, formed fewer AGEs than glucose. Methods. Hemoglobin (Hb), a counter-protein of AGE, was incubated in four kinds of medium; glucose-phosphate buffered saline (PBS), autoclaved glucose-PBS, trehalose-PBS, and autoclaved trehalose-PBS, for 3, 7, 14, and 30 days, respectively. Polymerization of the Hb molecule was detected by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and carboxymethylated Hb was detected by Western blotting, using specific mono-clonal antibody for carboxymethylated N-terminal valine-Hb (CMV-Hb). Results. PBS containing glucose showed bands of polymerized Hb molecule, a phenomenon which was markedly exaggerated by autoclaving. Likely, PBS containing glucose showed the formation of CMV-Hb in the long incubation of 30 days, and PBS containing autoclaved glucose showed accelerated formation of CMV-Hb in an incubation as short as 3 days. By contrast, PBS containing trehalose showed much less increase in a band of 30k Dalton and in CMV-Hb formation even in autoclaved medium. Conclusions. Our present in vitro study clearly showed the superior characteristic of trehalose to produce fewer AGEs. Based upon the results of this study, we propose that the application of trehalose should be considered for CAPD solution.     

Relation between lymphatic vessel diameter and clinicopathologic parameters in squamous cell carcinomas of the oral region.

BACKGROUND: In an attempt to determine the mechanism of cervical lymph node metastases, the authors studied the relation between lymphatic vessels in or around tumor tissue and lymph node metastases in patients with primary squamous cell carcinoma (SCC) of the oral region by enzyme histochemistry using 5'nucleotidase-alkaline phosphatase. METHODS: The subjects were 23 patients who had biopsy proven oral SCC. After enzyme histochemical staining, the cross-sectional dimension (referred to as diameter) of the lymphatic vessels were measured and analyzed in relation to the T classification of the tumor, degree of tumor differentiation, and mode of invasion. RESULTS: The average diameter of the lymphatic vessels in or around tumor tissue was significantly greater than that in tumor free tissue (P < 0.01). The mode of invasion correlated significantly with the lymphatic vessel diameter (P < 0.01). The diameter did not correlate significantly with the T classification (P range, 0.135-0.254) or tumor differentiation (P = 0.274). The following relation was found between the incidence of cervical lymph node metastases and the mode of invasion: 40.0% of Grade 2 tumors were positive for metastases, 71.4% of Grade 3 tumors were positive, and 75.0% of Grade 4 tumors were positive (grading was according to Jakobsson's classification). CONCLUSIONS: Of the factors evaluated in this study, only the mode of invasion correlated significantly with the diameter of the lymphatic vessels. Although other studies have shown that tumor thickness and perhaps even perineural and blood vessel invasion may be equally important, the findings of the current study suggest that both lymphatic vessel diameter and the mode of invasion may be important factors in the prediction of cervical lymph node metastases.     

Hyperpolarization-induced dilatation of submucosal arterioles in the guinea-pig ileum

1. The effects of inhibition of acetylcholine (ACh)-induced hyperpolarization on dilatation of submucosal arterioles were investigated in the guinea-pig ileum. 2. In smooth muscles of the arterioles depolarized by Ba(2+) (0.5 mM) to about -40 mV, ACh (3 microM) repolarized the membrane to about -65 mV (hyperpolarization), irrespective of the absence or presence of L-N(omega)-nitroarginine (L-NOARG, 0.1 mM) and diclofenac (1 microM), and increased the diameter (dilatation). 3. Combined application of charybdotoxin (CTX, 50 nM) and apamin (0.1 microM), inhibitors of some types of K(+)-channels, abolished the ACh-induced hyperpolarization and dilatation. 4. 18 beta-Glycerrhetinic acid (18 beta-GA, 30 microM), a known inhibitor of gap junctions, depolarized the membrane to about -36 mV, either in the absence or in the presence of Ba(2+), with no associated contraction of the arterioles. In the presence of 18 beta-GA, ACh-induced hyperpolarization was abolished, however the dilatation was inhibited only partially, with associated inhibition of constriction produced by Ba(2+) and NA. 5. 18 beta-GA inhibited the dilatation produced by sodium nitroprusside, an NO donor. 6. The ACh-induced hyperpolarization and dilatation were abolished in the presence of 2-aminoethoxydiphenyl borate (30 microM), an inhibitory modulator of inositol trisphosphate receptor-mediated Ca(2+) release from intracellular stores. 7. It is concluded that in submucosal arterioles, hyperpolarizations produced by ACh have causal relationship to the arteriolar dilatation. 18 beta-GA did not induce parallel relationship between hyperpolarization and dilatation produced by ACh. 18 beta-GA may have unidentified inhibitory effects on agonist-mediated actions, in addition to the inhibition of gap junctions.     

The volume limitation of the galeal temporalis flap in facial augmentation

Galeal temporalis flaps based on the superficial temporal vessels have been used for facial augmentation and can be extended to the contralateral side beyond the midline in order to achieve maximum volume. In five patients, the volumes of extended galeal flaps were measured intraoperatively using a water displacement method. The calculated volume varied between 28 and 38 cm3. Experience with this flap showed satisfactory results with no complications; therefore, it is concluded that the extended galeal temporalis flap may be a first choice in the correction of facial soft tissue deficits less than 40 cm3. Clinical cases are presented.     

Gastric Wall Structure Using a 30 MHz Endoscopic Ultrasound Probe,Focusing upon Delineation of the Muscularis Mucosae

Background: The normal gastric wall has been reported to appear to be a five‐layered structure. The structure of the gastric wall using a 30 MHz endoscopic ultrasound probe and especially the identification of the muscularis mucosae (MM), has not been analyzed clearly. Methods: In a basic study, 11 sections of normal gastric wall with 26 horizontally inserted nylon sutures were immersed in water. The sections were scanned and the findings correlated using standard histology. In a clinical study, 15 early gastric cancers were examined by a 30 MHz endoscopic ultrasound probe. Results: In a basic study, layers deeper than the lower part of the submucosa could not be seen using ultrasonography. The first to fourth layers represented the mucosal layer except the MM, the fifth layer (high‐echo layer) represented the boundary echo and a part of the MM, while the sixth layer (low‐echo layer) represented the rest of the MM. The muscularis mucosae was seen clearly in all samples. In a clinical study the layers deeper than the submucosal layer could not be seen and the MM was visible in 87% of cases. The depth of invasion was estimated accurately in 90% of mucosal cancers and in 80% of submucosal cancers. Conclusion: A 30 MHz endoscopic ultrasound probe, which cannot image the entire gastric wall, can visualize the MM and may help to confirm the structure of gastric wall layers and improve the ability to determine the depth of invasion in gastric cancer.     

Long-term results of Konno procedure for complex left ventricular outflow tract obstruction.

OBJECTIVE: The current study aims to evaluate the long-term outcomes of the Konno procedure. METHODS: The clinical records of 63 patients who had undergone the Konno procedure between February 1984 and March 2007 were reviewed. During this period, the Ross procedure was introduced in 1996. Of the 63 patients, 38 were male and 25, female. Their ages at the time of operation ranged from 1 year 9 months to 37 years, and their body weights ranged from 8.1 to 63 kg. Valves larger than 23 mm were used in 57 patients. RESULTS: There was one hospital death (myocardial infarction) and six late deaths (sudden death, 2; congestive heart failure, 2; infectious endocarditis, 1; traffic accident, 1). The Kaplan-Meier survival rates including hospital mortality and late mortality were 91.9% at 10 years and 87.7% at 15 years. There were 20 significant complications in 16 patients: thromboembolism was noted in 1 patient; reoperations (Konno procedure (aortic valve replacement), 5 (thrombosed valve, 3; pannus formation, 1; IE, 1); mitral valve replacement, 3; coronary artery bypass grafting, 2; grafting of the descending aorta, 1), balloon dilatation for recoarctation, and 7 catheter interventions were required in 9, 1, and 5 patients, respectively. The event-free rates including all events were 75.2% at 10 years and 67.2% at 15 years. In the long-term period, the results of echocardiography revealed good cardiac function. CONCLUSIONS: The Konno procedure is effective and safe for the treatment of complex left ventricular outflow tract obstruction and for the preservation of ventricular function. Since some issues concerning anticoagulation-related complications and infectious endocarditis remain, careful observation is mandatory.     

Influenza vaccine with Surfacten, a modified pulmonary surfactant, induces systemic and mucosal immune responses without side effects in minipigs

Immune responses and side effects of intranasally administered flu vaccine with the commercial product Surfacten, a modified bovine pulmonary surfactant, were investigated in minipigs. The use of minipigs was based on the anatomical resemblance of nasal lymph nodes, the principal antigen uptake site of respiratory mucosal immunity, between pig and human. Intranasal instillation of HA vaccine adjuvanted with Surfacten elicited significantly higher serum hemagglutination inhibition titers than the antigen alone, with wide cross-neutralizing activities of secretory IgA in nasal washes. No significant induction of inflammatory cytokines or migration of inflammatory cells was observed at the site of immunization or serum after the first immunization. These data suggest the potential usefulness of Surfacten for mucosal vaccination.     

Operative fiberoptic nephroureteroscope--removal of renal and upper ureteral stones

Two types of operative fiberoptic nephroureteroscopes were developed with the cooperation of the Olympus Optical Company mainly to remove the upper urinary tract calculi. Removal of the renal and upper ureteral stones was attempted in nine cases. The ureter was dilated with olive tip, balloon and Teflon dilators alone or in combination. The combination of balloon and Teflon dilators seemed the most promising. As a result of ureteral dilation, operative fiberoptic nephroureteroscopes, 4.5 mm and 3.5 mm in diameter, could be passed into the ureter in all the cases and the stones could be visualized clearly. Four of the 7 upper ureteral stones and 1 of the 2 pelvic stones could be removed. The success ratio was 56%. The method of stone removal still requires improvement. At present, application of the operative fiberoptic nephroureteroscope is indicated for upper ureteral and renal stones less than 1.0 cm in diameter. With the improved techniques of stone removal using this fiberscope, the indications of extracorporeal shock-wave lithotripsy or percutaneous nephrolithotripsy will probably be greatly reduced in the near future.     

Effects of inhibitors of nonselective cation channels on the acetylcholine-induced depolarization of circular smooth muscle from the guinea-pig stomach antrum.

In circular smooth muscle bundles isolated from the guinea-pig stomach antrum, the effects of quinidine, Ni2+, flufenamic acid, niflumic acid, La3+, SKF-96365 and 4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) on acetylcholine (ACh)-induced depolarization were investigated. Recording membrane potentials from smooth muscle cells with intracellular microelectrodes revealed that ACh (1 microM) depolarized the membrane by 5-8 mV and increased the amplitude and frequency of slow potentials. These effects were inhibited by atropine. Quinidine (10 microM) increased the amplitude of ACh-induced depolarization, with no alteration to the properties of slow potentials. Ni2+ (50 microM) transiently (5-10 min) depolarized the membrane by about 5 mV, with an associated increase in frequency and amplitude of slow potentials. In the stabilized condition with Ni2+, the amplitude of ACh-induced depolarization remained unchanged. Flufenamic acid (10 microM) inhibited the generation of slow potentials, with no change in either the amplitude of ACh-induced depolarization or of the amplitude and frequency of slow potentials generated during ACh stimulation. A high concentration of flufenamic acid (100 microM) depolarized the membrane and increased the amplitude of ACh-induced depolarization. Niflumic acid (10 microM) hyperpolarized the membrane and increased the amplitude and frequency of slow potentials and also the amplitude of ACh-induced depolarization. DIDS (100 microM) hyperpolarized the membrane and inhibited the amplitude and frequency of slow potentials, with no alteration to the amplitude of ACh-induced depolarization. SKF-96365 (3-50 microM) depolarized the membrane in a concentration-dependent manner, but did not change the level of ACh-induced depolarization. La3+ (50 microM) did not alter the properties of the slow potentials or the ACh-induced responses. These results provide evidence that ACh-induced depolarization is not inhibited by chemicals known to inhibit non-selective cation channels. We suggest that muscarinic receptor-mediated signal transduction may be different in smooth muscle and interstitial cells.