The Geriatric Nutritional Risk Index is Independently Associated with Prognosis in Patients with Critical Limb Ischemia Following Endovascular Therapy.

OBJECTIVES: Patients with critical limb ischemia (CLI) have poor overall and limb prognosis. Although nutritional status influences overall prognosis, and the Geriatric Nutritional Risk Index (GNRI) is a widely used, simple and well established nutritional status screening method, the association between the GNRI and the overall and limb prognosis of patients with CLI following endovascular therapy (EVT) has not been explored. METHODS: Clinical outcomes were retrospectively evaluated in 473 consecutive patients (74 ± 10 years; 59% male) with CLI who underwent EVT. The GNRI on admission was calculated as follows: [14.89 × albumin (g/dL)] + [41.7 × (body weight/ideal body weight)]. Cox proportional hazard analysis was performed to explore the independent association between the GNRI and mortality and major amputation. RESULTS: Patients (53% ambulatory, 38% wheelchair bound, and 9% bedridden) were divided into two groups based on the median GNRI: the higher group (GNRI ≥ 91.2, n = 237) and the lower group (GNRI < 91.2, n = 236). Median follow up duration after EVT was 11.3 months. Three years after EVT, the survival rate (74% in the higher GNRI, and 48% in the lower GNRI, respectively), and limb salvage rate (92% in the higher GNRI, and 84% in the lower GNRI) were significantly lower in the lower GNRI group. GNRI (hazard ratio [HR], 1.03; 95% confidence interval [CI], 1.01-1.05), along with being wheelchair bound (HR, 1.87; 95% CI 1.17-2.97; vs. ambulatory status), being bedridden (HR, 3.10; 95% CI, 1.63-2.97; vs. ambulatory status), being on hemodialysis (HR, 2.33; 95% CI, 1.49-3.64), and having chronic heart failure (HR, 2.22; 95% CI, 1.44-3.43) were the independent predictors of mortality. The GNRI (HR, 1.04; 95% CI, 1.01-1.07), being bedridden (HR, 4.15; 95% CI, 1.67-10.3; vs. ambulatory status), isolated below knee disease (HR, 2.49; 95% CI, 1.30-4.77), and hemodialysis (HR, 2.44; 95% CI, 1.23-4.85) were independently associated with major amputation. CONCLUSIONS: The GNRI on admission was independently associated with mortality and major amputation after EVT in patients with CLI.

Predictors of delayed wound healing after endovascular therapy of isolated infrapopliteal lesions underlying critical limb ischemia in patients with high prevalence of diabetes mellitus and hemodialysis.

OBJECTIVES: Acceptable limb salvage rates underlie the widespread use of endovascular therapy (EVT) for patients with critical limb ischemia (CLI) secondary to isolated infrapopliteal lesions; however, post-EVT delayed wound healing remains a challenge. Predictors of delayed wound healing and their use in risk stratification of EVT in patients with CLI due to isolated infrapopliteal lesions are explored. METHODS: This was a retrospective multicenter study. 871 consecutive critically ischemic limbs were studied. There was tissue loss in 734 patients (age: 71 ± 10 years old; 71% male) who had undergone EVT between April 2004 and December 2012. The wound healing rate after EVT was estimated by the Kaplan-Meier method. The association between baseline characteristics and delayed wound healing was assessed by the Cox proportional hazard model. RESULTS: Diabetes mellitus and regular dialysis were present in 75% (553/734) and 64% (476/734) of patients, respectively; 67% of limbs (585/871) had Rutherford class 5 CLI; 8% (67/871) of wounds were located in the heel only; 25% (219/871) of limbs had Rutherford 6 (involving not only the heel); and 42% (354/871) of wounds were complicated by infection. The rate of freedom from major amputation at 1 year reached 88%, whereas the wound healing rate was 67%. Median time to wound healing was 146 days. By multivariate analysis, non-ambulatory status (hazard ratio [HR], 1.58; 95% confidence interval [CI] 1.31-1.91) serum albumin <3 g/dL (HR 1.42; 95% CI 1.08-1.86), Rutherford 6 (not only heel) (HR 1.68; 95% CI 1.33-2.14), wound infection (HR 1.24; 95% CI 1.03-1.50), EVT not based on angiosome concept (HR 1.28; 95% CI 1.06-1.55), and below the ankle (BTA) 0 vessel runoff after EVT (HR 1.45; 95% CI 1.14-1.86) were independent predictors of delayed wound healing. CONCLUSIONS: Non-ambulatory status, low albumin level, Rutherford 6 (not only heel), wound infection, indirect intervention, and poor BTA runoff were independent predictors for delayed wound healing after EVT in patients with CLI secondary to infrapopliteal lesions, and their use in risk stratification allows estimation of the wound healing rate.

Ectopic expression of cone-specific G-protein-coupled receptor kinase GRK7 in zebrafish rods leads to lower photosensitivity and altered responses.

To investigate the roles of G-protein receptor kinases (GRKs) in the light responses of vertebrate photoreceptors, we generated transgenic zebrafish lines, the rods of which express either cone GRK (GRK7) or rod GRK (GRK1) in addition to the endogenous GRK1, and we then measured the electrophysiological characteristics of single-cell responses and the behavioural responses of intact animals. Our study establishes the zebrafish expression system as a convenient platform for the investigation of specific components of the phototransduction cascade. The addition of GRK1 led to minor changes in rod responses. However, exogenous GRK7 in GRK7-tg animals led to lowered rod sensitivity, as occurs in cones, but surprisingly to slower response kinetics. Examination of responses to long series of very dim flashes suggested the possibility that the GRK7-tg rods generated two classes of single-photon response, perhaps corresponding to the interaction of activated rhodopsin with GRK1 (giving a standard response) or with GRK7(giving a very small response). Behavioural measurement of optokinetic responses (OKR) in intact GRK7-tg zebrafish larvae showed that the overall rod visual pathway was less sensitive, in accord with the lowered sensitivity of the rods. These results help provide an understanding for the molecular basis of the electrophysiological differences between cones and rods.

Coexpression of microsomal-type prostaglandin E synthase with cyclooxygenase-2 in brain endothelial cells of rats during endotoxin-induced fever.

Fever is triggered by an elevation of prostaglandin E(2) (PGE(2)) in the brain. However, the mechanism of its elevation remains unanswered. We herein cloned the rat glutathione-dependent microsomal prostaglandin E synthase (mPGES), the terminal enzyme for PGE(2) biosynthesis, and examined its induction in the rat brain after intraperitoneal injection of pyrogen lipopolysaccharide (LPS). In Northern blot analysis, mPGES mRNA was weakly expressed in the brain under the normal conditions but was markedly induced between 2 and 4 hr after the LPS injection. In situ hybridization study revealed that LPS-induced mPGES mRNA signals were mainly associated with brain blood vessels, especially vein or venular-type ones, in the whole brain area. Immunohistochemical study demonstrated that mPGES-like immunoreactivity was expressed in the perinuclear region of brain endothelial cells, which were identified as von Willebrand factor-positive cells. Furthermore, in the perinuclear region of the endothelial cells, mPGES was colocalized with cyclooxygenase-2 (COX-2), which is the enzyme essential for the production of the mPGES substrate PGH(2). Inhibition of cyclooxygenase-2 activity resulted in suppression of both PGE(2) level in the CSF and fever (Cao et al., 1997), suggesting that the two enzymes were functionally linked and that this link is essential for fever. These results demonstrate that brain endothelial cells play an essential role in the PGE(2) production during fever by expressing COX-2 and mPGES.

Genetic transfer of endothelin converting enzyme activity to CHO-K1 cells: detection of positive cells by reverse hemolytic plaque assay.

We have established a novel method of molecular cloning of endothelin converting enzyme, a key enzyme in the production of a potent vasoconstrictor endothelin-1, by modification of the reverse hemolytic plaque assay. Also, we demonstrated that a cell line, CHO-K1, showed no detectable activity of endothelin converting enzyme. This cell line was transfected with a cDNA library of bovine endothelial cells. The modified reverse hemolytic plaque assay was shown to detect even a single CHO-K1 cell that was changed to produce mature ET-1 by transfection. Thus, this novel method is suggested to be useful for the molecular cloning of other secreted antigens and their processing enzyme.

Cloning full-length, cap-trapper-selected cDNAs by using the single-strand linker ligation method.

We have developed the single-strand linker ligation method (SSLLM), which uses DNA ligase to add a dsDNA linker to single-stranded (ss) full-length cDNA. The linkers have random 6-bp (dN6 or dGN5) 3' overhangs that can ligate to any cDNA sequence, thereby facilitating the production of cDNA libraries with titers exceeding 1 x 10(6) independent clones. We confirmed that the 5' ends of cDNA inserts cloned by using SSLLM are full-length and include the 5' untranslated regions. The great advantage of our method is that the elimination of the GC tail simplifies the sequencing and protein translation of the full-length clones. Further, our method tags ss cDNAs more efficiently than does the traditional RNA ligase reaction.

Removal of polyA tails from full-length cDNA libraries for high-efficiency sequencing.

We have developed a method to overcome sequencing problems caused by the presence of homopolymer stretches, such as polyA/T, in cDNA libraries. PolyA tails are shortened by cleaving before cDNA cloning with type IIS restriction enzymes, such as GsuI, placed next to the oligo-dT used to prime the polyA tails of mRNAs. We constructed four rice Cap-Trapper-selected, full-length normalized cDNA libraries, of which the average residual polyA tail was 4 bases or shorter in most of the clones analyzed Because of the removal of homopolymeric stretches, libraries prepared with this method can be used for direct sequencing and transcriptional sequencing without the slippage observed for libraries prepared with currently available methods, thus improving sequencing accuracy, operations, and throughput.

A p53 polymorphism associated with increased risk of hepatitis C virus infection.

Incidence of polymorphisms on exon 4 (CGC vs. CCC, p53Arg vs. p53Pro, A2 allele vs. A1 allele at codon 72, respectively) of the p53 gene was compared in 75 cases with hepatitis C virus (HCV) infection and 232 noninfected control subjects in a defined geographical area in Japan. Polymorphism was analyzed by the polymerase chain reaction-single strand conformation polymorphism method using DNA from peripheral blood leukocytes. When all cases and controls were compared, there was no significant correlation between hepatitis C virus (HCV) infection and the p53 polymorphism in question. However, when male cases infected with HCV type 1b, the most common viral genotype, were compared with controls matched by sex and age, significantly higher homozygotes for p53Pro were found in cases compared with controls (P=0.039). Significantly higher allelic frequency of this polymorphism was also observed with cases (P=0.010). We found no significant statistical difference between the p53 polymorphism and other genotypes of HCV (2a, 2b and others). On the basis of our study we believe there exists a significant correlation between male homozygotes for p53Pro with HCV type 1b infection.

LiV2O4 spinel as a heavy-mass fermi liquid: anomalous transport and role of geometrical frustration

Transport and specific heat measurements on hydrothermally grown single crystals reveal the formation of a heavy-mass Fermi liquid in the LiV2O4 spinel, below a coherence temperature of T* = 20-30 K. A few observations which illustrate the uniqueness of this spinel are discussed in connection with the origin of the heavy mass, such as the anomalous absence of resistivity saturation above T* and the close proximity to a spin glass phase where the influence of the magnetic frustration is evident.

Antihypertensive effects of CS-045 treatment in obese Zucker rats.

The association of hypertension with obesity has been well recognized, but the etiology remains poorly understood. Obesity is characterized by hyperinsulinemia, which reflects peripheral insulin resistance. Insulin resistance may participate in the development of hypertension with obesity. CS-045 [(I)-5-[4-(5-hydroxy-2,5,7,8-tetramethylchroman-2-yl-methoxy)be nzy l]-2,4- thiazolidiendion] is a new orally effective antidiabetic agent that potentiates insulin action and reduces insulin resistance in obese Zucker rats and other obese diabetic animals. In this study, we examined the antihypertensive effect of CS-045 in obese male and female Zucker rats as a model of hypertension associated with obesity. CS-045 was administered as a food admixture (approximately 16 and approximately 70 mg/kg/d) for 4 weeks (female) and (approximately 15 and approximately 67 mg/kg/d) 8 weeks (male) in obese Zucker rats at 5 to 7 months of age. CS-045 slightly but significantly decreased plasma glucose levels. Plasma insulin levels were significantly decreased in obese male rats, but were not significantly decreased in obese female rats. Drug administration led to significant decreases in plasma triglyceride and cholesterol levels and systolic blood pressure (SBP) in a dose-dependent manner from 1 week after administration in obese Zucker rats. CS-045 increased urinary sodium excretion, sodium/potassium ratio, and creatine clearance in a dose-dependent manner, and also led to a remarkable decrease in urinary protein excretion. However, CS-045 did not reduce urinary catecholamine excretion. These data indicate that CS-045 may promote renal sodium excretion and improve decreased glomerular filtration rates, which may reflect the amelioration of insulin resistance.(ABSTRACT TRUNCATED AT 250 WORDS)

l-Menthol-induced [Ca2+]i increase and impulses in cultured sensory neurons.

We investigated the effects of l-menthol on cultured dorsal root ganglion (DRG) cells, instead of free nerve endings of sensory fibers. Using Fura-2 microfluorimetry, we identified a few DRG neurons that showed an increase in intracellular free Ca2+ concentration ([Ca2+]i) in response to l-menthol. They made up only 10% of the neurons activated by a high K+ solution. l-Menthol induced the [Ca2+]i increase in a dose-dependent manner, with an EC50 of 37.9 microM and a Hill coefficient of 0.97. A related compound, cyclohexanol, had no effect. When extracellular Ca2+ was removed, l-menthol did not induce the [Ca2+]i increase. Whole-cell current-clamp recordings revealed that l-menthol induced depolarization (13.2 mV, receptor potential) leading to impulses. We conclude that l-menthol induced the impulses through activation of menthol receptors in a small subset of the cultured sensory neurons.

Prostaglandin E2 excites neurons of the nucleus tractus solitarius by activating cation channels.

Nucleus tractus solitarius (NTS) has a high density of prostaglandin E2 (PGE2)-binding sites. Action of PGE2 (10(-9)-10(-6) M) was tested on neurons in a NTS slice with patch-clamp recording under synaptic blockade. PGE2 raised the firing rate in approximately half of the neurons in cell-attached patch mode. In whole-cell current clamp, PGE2 depolarized membrane potential accompanied by an increase in firing rate. In whole-cell voltage clamp (-58 mV), PGE2 induced the inward current with an increase in conductance. The current was linearly related to voltage from -100 mV to -10 mV and suppressed between -10 mV and 20 mV. The current-voltage curve remained similar under low external Cl- or high internal Cl- conditions and after external Na+ was replaced by Cs+. It is concluded that PGE2 excites NTS neurons by activating cation conductance.

Effects of emergency coronary angioplasty on left ventricular volume after acute myocardial infarction compared with intracoronary thrombolysis.

BACKGROUND: The effects of emergency coronary angioplasty on left ventricular remodeling have not been fully evaluated. We compared the effects of emergency coronary angioplasty on left ventricular volume with those of intracoronary thrombolysis in patients suffering from their first acute myocardial infarction (AMI). METHODS: A total of 51 patients who had a culprit lesion at site No 6 of the American Heart Association (AHA) classification were analyzed. Patients with re-obstruction of the culprit lesion after reperfusion were excluded from the study. The coronary angioplasty and the thrombolysis groups consisted of 28 and 23 patients, respectively. Left ventricular volume was measured by left ventriculography at 4 weeks after infarction. RESULTS: Clinical and angiographic background factors showed no significant differences between these two groups. The left ventricular end-systolic volume index was significantly smaller in the angioplasty group than in the thrombolysis group (31 +/- 14 versus 45 +/- 14 ml/m2, P < 0.01). Similarly, the left ventricular end-diastolic volume index was significantly smaller in the angioplasty group (64 +/- 13 versus 82 +/- 13 ml/m2, P < 0.01). Conversely, the left ventricular ejection fraction was larger in the angioplasty group than in the thrombolysis group (54 +/- 13 versus 45 +/- 13%, P < 0.05). Multivariate analysis revealed that the selection of coronary angioplasty was one of the factors significantly associated with a decrease in left ventricular end-systolic and end-diastolic volume. CONCLUSION: These results suggest that emergency coronary angioplasty produces better left ventricular remodeling than intracoronary thrombolysis after AMI.

Prediction of tableting problems such as capping and sticking: theoretical calculations.

Tabletability factors were developed through the use of an automated tabletability tester, and the applicability of these factors to actual manufacturing procedures was tested by the large-scale manufacturing of tablets. Use of this approach does not require knowledge of the tablet's physical properties, e.g., moisture content, porosity, particle size, fluidity, and angle of repose.

Cyclooxygenase in the vagal afferents: is it involved in the brain prostaglandin response evoked by lipopolysaccharide?

The vagal afferents are proposed to transmit abdominal immune signals to the brain. In this immune-brain communication, prostaglandins might play a mediator role. In fact, prostaglandin receptors are abundant in the vagal afferents. We examined here the presence of cyclooxygenase, an enzyme necessary for prostaglandin biosynthesis, in the vagal afferents of rats. We also tested whether the vagal afferents contribute to the elevation of prostaglandin E2 in the brain after intraperitoneal injection of lipopolysaccharide. Under normal conditions, cyclooxygenase-1-like immunoreactivity was constitutively expressed in the vagal afferents at their central terminals and in their cell bodies. Cyclooxygenase-2-like immunoreactivity was absent in the vagal afferents under normal as well as lipopolysaccharide-challenged conditions. Instead, cyclooxygenase-2-like immunoreactivity was induced in brain endothelial cells by the lipopolysaccharide challenge. The elevation of prostaglandin E2 in the cerebrospinal fluid after lipopolysaccharide challenge was not inhibited, but was rather enhanced, by the bilateral vagotomy. These results suggest that the vagal afferents potentially generate prostaglandins, which may locally modulate the vagal signal transmission, but that the vagal afferents are not essential to the elevation of prostaglandin E2 in the brain after intraperitoneal challenge with LPS.

A case-control study of hepatocellular carcinoma in Hyogo Prefecture.

We conducted a case-control study to elucidate factors contributing to the development of hepatocellular carcinoma (HCC) in Hyogo Prefecture, Japan where its incidence is high. In this study, the incidence of hepatitis virus infection in HCC patients and lifestyle factors including dietary and drinking habits were investigated. One hundred and eight patients with HCC (89 males and 19 females) and 93 controls (63 males and 30 females) were enrolled. Hepatitis C virus (HCV) antibody was identified in 87% of cases of which HCV-RNA was further detected in more than 95%. Genotype distribution of HCV was similar to that reported for the Japanese infected with HCV. There was a positive correlation between HCC and history of blood transfusion (both male and female) and surgery (male). Alcohol consumption over a period of 30 years was higher in the HCC patients than in controls; but not significant. These data indicated that HCV infection is a primary contributing factor to the development of HCC in Hyogo Prefecture.

Bone and soft tissue tumours of the foot: review of 83 cases.

PURPOSE: To review cases of bone and soft tissue tumours of the feet managed at the Hyogo College of Medicine, Nishinomiya and Takarazuka Municipal Hospital, Takarazuka, Japan. METHODS: Retrospective analysis of 83 patient records treated for bone and soft tissue tumours of the feet between 1974 and 2000. RESULTS: There were 33 benign bone tumours, one primary malignant bone tumour, and 2 metastatic bone cancer. Marginal resection was performed in cases of osteochondroma and curettage in cases of other benign bone tumour. Despite below-knee amputation in the case of chondrosarcoma, the patient died because of pulmonary metastasis. Two patients with metastatic cancer also died, and 2 cases of osteochondroma and one of benign chondroblastoma recurred. There were 47 cases of soft tissue tumour. Treatment for benign soft tissue tumours was marginal resection; no cases recurred. In contrast, all patients with soft tissue sarcoma died after surgery. The majority of bone tumours were located in the toe and hindfoot areas, in the first and second decades of life, whereas soft tissue tumours occurred mainly in the midfoot area and in patients aged between 20 and 50 years. The sex distribution was almost even for bone tumours (male: female ratio, 19:17), whereas about half as many males as females had soft tissue tumours (14:33). CONCLUSION: Bone and soft tissue tumours of the feet are uncommon. Most bone tumours are chondrogenic, but differential diagnosis of malignant from benign disease is difficult and prognosis is poor. Management of benign tumours by marginal resection has good prognosis, whereas prognosis of soft tissue sarcomas is very poor.

[Studies on faint attack due to arrhythmias with Holter electrocardiogram].

Out of 1,400 Holter ECGs, 15 records revealed arrhythmias which would be responsible for faint attack, including 8 sick sinus syndrome with cardiac arrest, 4 non-sustained ventricular tachycardia (VT), 2 paroxysmal atrial fibrillation and 1 paroxysmal supraventricular tachycardia. All of cardiac arrest corresponding to faint attack lasted for longer than 4 sec, and an averaged duration of the arrest was 5.8 +/- 2.3 sec. This was significantly longer than that with no subjective symptoms, 3.4 +/- 0.7 sec. Duration and rate of VT did not show any significant relationship with faint attack. The result suggests that cardiac arrest is mostly responsible for arrhythmia-induced involvement of cerebral circulation and faint attack. It is also suggested that cardiac arrest longer than 4 sec should be extensively treated including pacemaker implantation.

[Giant plasmacytoma of the skull which appeared in the clinical course of multiple myeloma--a case report].

A 51-year-old woman with large plasmacytoma occurring from the temporal bone is presented. She has a history of multiple myeloma for 9 years. She manifested marked swelling in the left temporal area with tenderness. Neurological examination revealed no abnormality. She showed monoclonal free light chain (lambda type) in the serum and urine, and had multiple osteolytic lesions in her general bones. T1 WI of MRI exhibited a huge mass showing slightly high intensity in the left middle fossa and infratemporal fossa, and a part of the mass protruded into the extracranial space. The mass was markedly enhanced by Gd-DTPA. Angiography showed a hypervascular mass supplied by the external carotid artery. Biopsy disclosed plasmacytoma. She underwent local irradiation of 30 Gy and chemotherapy of Ranimustine (100 mg) and Cyclophosphamide (400 mg). The tumor reduced its size, and tenderness in her temporal area disappeared.

[A malignant thymoma combined with sarcoidosis].

A case of 69-year-old female of malignant thymoma with sarcoidosis was experienced. Although many diseases such as myasthenia gravis are associated with thymoma, sarcoidosis is a rare thymoma-associated disease. Only one case of thymoma with sarcoidosis was reported all over the world, and our case is the first case in Japan. As thymus is essential for the development of T lymphocytes, thymoma may cause T lymphocytes disorder and sarcoidosis. So, we consider thymoma with sarcoidosis as a syndrome rather than as a spectrum of coincidental disease.