Unusual diagnosis of feline cardiac lymphoma using cardiac needle biopsy.

BACKGROUND: Cardiac tumors in cats are relatively rare, with lymphoma accounting for more than half of all cases. However, feline cardiac lymphoma is often diagnosed post-mortem, and it is difficult to diagnose while the cat is still alive. It is the first report of a direct, rather than estimative, diagnosis with cardiac needle biopsy of a living cat with cardiac lymphoma. CASE PRESENTATION: A 3-year-old domestic short-haired male cat experienced loss of energy and loss of appetite. Thoracic radiography and transthoracic echocardiography showed cardiomegaly with slight pleural effusion and cardiac tamponade due to pericardial effusion, respectively. In addition, partial hyperechoic and hypertrophy of the papillary muscle and myocardium were observed. Blood test showed an increase in cardiac troponin I levels. Pericardial fluid, removed by pericardiocentesis, was analyzed; however, the cause could not be determined. With the owner's consent, pericardiectomy performed under thoracotomy revealed a discolored myocardium. Cardiac needle biopsy was performed with a 25G needle, and a large number of large atypical lymphocytes were collected; therefore, a direct diagnosis of cardiac lymphoma was made. Pathological examination of the pericardium diagnosed at a later date revealed T-cell large cell lymphoma. The cat underwent chemotherapy followed by temporary remission but died 60 days after the diagnosis. Postmortem, two-dimensional speckle-tracking echocardiography (data when alive) revealed an abnormal left ventricular myocardial deformation, which corresponded to the site of cardiac needle biopsy. CONCLUSIONS: This rare case demonstrates that cardiac lymphoma should be added to the differential diagnosis in cats with myocardial hypertrophy and that the diagnosis can be made directly by thoracotomy and cardiac needle biopsy. In addition, the measurement of cardiac troponin I levels and local deformation analysis of the myocardium by two-dimensional speckle-tracking echocardiography may be useful in the diagnosis of cardiac tumors.

Enhanced exudation of malate in the rhizosphere due to AtALMT1 overexpression in blackgram (Vigna mungo L.) confers increased aluminium tolerance.

Worldwide, 50% of soil is acidic, which induces aluminium (Al) toxicity in plants, as the phyto-availability of Al3+ increases in acidic soil. Plants responds to Al3+ toxicity by exuding organic acids into the rhizosphere. The organic acid responsible for Al3+ stress response varies from species to species, which in the case of blackgram (Vigna mungo L.) is citrate. In blackgram, an Arabidopsis malate transporter, AtALMT1, was overexpressed with the motive of inducing enhanced exudation of malate. Transgenics were generated using cotyledon node explants through Agrobacterium tumefaciens-mediated transformation. The putative transgenics were initially screened by AtALMT1-specific genomic DNA PCR, followed by quantitative PCR. Two independent transgenic events were identified and functionally characterized in the T3 generation. The transgenic lines, Line 1 and 2, showed better root growth, relative water content and chlorophyll content under Al3+ stress. Both lines also accounted for less oxidative damage, due to reduced accumulation of ROS molecules. Photosynthetic efficiency, as measured in terms of Fv /Fm , NPQ and Y(II), increased when compared to the wild type (WT). Relative expression of genes (VmSTOP1, VmALS3, VmMATE) responsible for Al3+ stress response in blackgram showed that overexpression of a malate transporter did not have any effect on their expression. Malate exudation increased whereas citrate exudation did not show any divergence from the WT. A pot stress assay found that the transgenics showed better adaptation to acidic soil. This report demonstrates that the overexpression of a malate transporter in a non-malate exuding species improves adaptation to Al3+ toxicity in acidic soil without effecting its stress response mechanism.

Carrier and spin dynamics of high-density exciton magnetic polarons in Cd0.8Mn0.2Te.

We investigated the carrier and spin dynamics of high-density exciton magnetic polarons (HD-EMPs) in Cd0.8Mn0.2Te based on the measurement of their time-resolved photoluminescence (PL) spectra and polarization states, and the utilization of photo-induced Faraday rotation techniques. The PL from the HD-EMPs were collected in a forward scattering configuration, and was observed as a pulsed emission of a few picoseconds duration, exhibiting a blue-shift with time evolution. The blue shift originated from the refractive-index dispersion of the sample. By excluding the influence of the refractive-index dispersion on the time profile, it was revealed that the ultra-short pulsed emission with a time width smaller than 1 ps was initially radiated with a time delay of ~2.4 ps after photoexcitation. From the results of time evolution of the polarization states, it is concluded that the exciton-Mn spin interactions occurs immediately after the excitation, which causes the Mn ion spins to align to follow the spin states of photoexcited excitons. The alignment of the Mn ion spins through the formation of the HD-EMPs was significantly faster than that of the localized EMP. On the other hand, the time evolution of the photo-induced Faraday rotation showed two decay components attributed to spin relaxations of the excitons and Mn ions within the HD-EMP. The observation of the Faraday rotation signal due to the Mn ion spins further confirms that these spins were aligned by the photo-excited spin-aligned excitons. Our findings suggest a novel mechanism for the effective optical control of spins in a semimagnetic semiconductor, which is associated with a multi-exciton system and its localized state.

GhSTOP1, a C2H2 type zinc finger transcription factor is essential for aluminum and proton stress tolerance and lateral root initiation in cotton.

Aluminum (Al) and proton (H+ ) ions are major acid soil stress factors deleteriously affecting plant root growth and crop yield. In our preliminary studies, cotton (Gossypium hirsutum L.) seedlings displayed very sensitive phenotypes to Al and H+ rhizotoxicities. Based on previous Arabidopsis results, we aimed to characterise the Al stress responsive Sensitive to Proton rhizotoxicity 1 (GhSTOP1) transcription system in cotton using RNAi-mediated down-regulation. With the help of seed embryo apex explants, we developed transgenic cotton plants overexpressing a GhSTOP1-RNAi cassette with NPTII selection. Kanamycin-tolerant T1 seedlings were further considered for Al and H+ stress tolerance studies. Down-regulation of the GhSTOP1 increased sensitivity to Al and proton rhizotoxicities, and root growth was significantly reduced in RNAi lines. The expression profile of GhALMT1 (Aluminum-activated Malate Transporter 1), GhMATE (Multidrug and Toxic Compound Extrusion), GhALS3 (Aluminium Sensitive 3) and key genes involved in the GABA shunt were down-regulated in the transgenic RNAi lines. Additionally, the lateral root initiation process was delayed and expression of GhNAC1, which is involved in lateral roots, was also suppressed in transgenic lines. Besides, overexpression of GhSTOP1 in Arabidopsis accelerated root growth and AtMATE and AtALMT1 expression under Al stress conditions. These analyses indicate that GhSTOP1 is essential for the expression of several genes which are necessary for acid soil tolerance mechanisms and lateral root initiation.

Usefulness of dyssynchrony indices based on two-dimensional speckle tracking echocardiography in a canine model of left bundle branch block.

INTRODUCTION: Determine the usefulness of dyssynchrony indices derived from two-dimensional speckle tracking echocardiography for the detection of mechanical dyssynchrony in a canine model of left bundle branch block. ANIMALS: Ten healthy beagles. MATERIALS AND METHODS: Segmental, time-radial strain curves were obtained using two-dimensional speckle tracking echocardiography. The maximum difference and standard deviation of the time to peak radial strain for six predefined segments (MaxD-TpSR and 6SD-TpSR) were calculated, together with the left ventricular dyssynchrony by radial strain (DysSR), before and after ablation of the left bundle branch block. Receiver operating characteristic curve analysis was performed using dogs after ablation as positive controls. RESULTS: After ablation, all dogs showed multiple peaks in at least one segment on the time-radial strain curve, while all dyssynchrony indices increased significantly (MaxD-TpSR from 16.25 ± 16.04 [mean ± standard deviation] to 44.4 ± 26.18 ms, 6SD-TpSR from 7.59 ± 7.40 to 19.62 ± 11.91 ms, and DysSR from 4.20 ± 2.12 to 10.87± 2.92%, p<0.05). In receiver operating characteristic curve analysis, the areas under the curve for MaxD-TpSR, 6SD-TpSR, and DysSR were 0.825, 0.800, and 0.980, respectively. CONCLUSIONS: Left ventricular dyssynchrony by radial strain can detect mechanical dyssynchrony with higher sensitivity and specificity than dyssynchrony indices, based on the time to peak radial strain.

Myocardial torsional deformations in cats with hypertrophic cardiomyopathy using two-dimensional speckle-tracking echocardiography.

OBJECTIVES: The objective of our study was to quantitatively measure systolic torsional deformations in cats with hypertrophic cardiomyopathy (HCM) and in controls. ANIMALS: Twenty-six client-owned cats with HCM and 14 healthy cats. HCM cats were categorized based on their symptoms (asymptomatic and symptomatic) and with or without left ventricular outflow tract obstruction (obstructive and non-obstructive). METHODS: The cats were examined for myocardial deformations using two-dimensional speckle-tracking echocardiography and were evaluated for peak systolic rotation and the rotation rate at each basal and apical view. Cats were also evaluated for the peak systolic torsion and torsion rate. RESULTS: The peak systolic apical rotation and torsion were higher in asymptomatic and symptomatic cats with HCM than in control cats. Also, the peak systolic apical rotation, apical rotation rate, torsion, and torsion rate were higher in cats with obstructive HCM than in control cats. CONCLUSIONS: Myocardial torsional deformations assessed by two-dimensional speckle-tracking echocardiography may be useful for evaluating compensatory myocardial function of HCM.

Increased undercarboxylated osteocalcin/intact osteocalcin ratio in patients undergoing hemodialysis.

UNLABELLED: Serum undercarboxylated osteocalcin (ucOC)/intact osteocalcin (iOC) ratio increased >1.0 in the patients undergoing hemodialysis, particularly in those with high bone turnover state. Consequently, serum ucOC/iOC ratio might lose its significance as a bone metabolic marker to indicate vitamin K deficiency in hemodialysis patients. INTRODUCTION: Serum intact osteocalcin (iOC), undercarboxylated OC (ucOC), and the ucOC/iOC ratio are considered clinically relevant indices in pre-dialysis chronic kidney disease (CKD) and hemodialysis (HD) patients, despite their accumulation in uremic serum. METHODS: Serum iOC and ucOC were measured along with serum intact parathyroid hormone (iPTH), bone alkaline phosphatase (BAP), and tartrate-resistant acid phosphatase (TRACP)-5b in 89 pre-dialysis CKD and 189 HD patients. RESULTS: Serum iOC and ucOC showed significantly negative correlations with estimated glomerular filtration rate in pre-dialysis CKD patients, although serum ucOC/iOC ratio did not correlate. Serum ucOC was significantly greater in HD patients than in pre-dialysis CKD patients, while serum iOC did not differ significantly, resulting in serum ucOC/iOC ratio >1.0 in 135 (71.4%) out of 189 HD patients. HD patients with high serum ucOC/iOC ratio (>1.0) had a significantly younger age and significantly higher values of body mass index, serum creatinine, albumin, phosphate, iPTH, and TRACP-5b than those with low ucOC/iOC ratio (≤ 1.0). The baseline iPTH and P1NP correlated with the changes of the ucOC/iOC ratio during the 2 days of the inter-dialytic period. Multivariate analysis showed that log [ucOC/iOC] in HD patients was significantly associated with log [iPTH], log [BAP], or log [TRACP-5b]. CONCLUSIONS: Serum ucOC/iOC ratio >1.0 was observed in as high as 71.4% of HD patients, preferentially with high bone turnover state, in comparison with pre-dialysis CKD patients. These data suggested that serum ucOC/iOC ratio might lose its significance as a bone metabolic marker to indicate vitamin K deficiency in HD patients.

Diffusion-weighted imaging vs STIR turbo SE imaging: capability for quantitative differentiation of small-cell lung cancer from non-small-cell lung cancer.

OBJECTIVE: To compare the capability of differentiation of small-cell lung cancer (SCLC) from non-SCLC (NSCLC) between diffusion-weighted imaging (DWI) and short tau inversion recovery (STIR) turbo spin-echo imaging. METHODS: The institutional review board of Kobe University Hospital, Kobe, Japan, approved this study, and written informed consent was obtained from each patient. 49 patients with NSCLC (30 males and 19 females; mean age, 66.8 years) and 7 patients with SCLC (5 males and 2 females; mean age, 68.6 years) enrolled and underwent DWI and STIR. To quantitatively differentiate SCLC from NSCLC, apparent diffusion coefficient (ADC) values on DWI and contrast ratios (CRs) between cancer and muscle on STIR were evaluated. ADC values and CRs were then compared between the two cell types by Mann-Whitney's U-tests, and the diagnostic performances were compared by McNemar's test. RESULTS: There were significant differences of mean ADC values (p < 0.001) and mean CRs (p = 0.003). With adopted threshold values, the specificity (85.7%) and accuracy (85.7%) of DWI were higher than those of STIR (specificity, 63.3%; p = 0.001 and accuracy, 66.1%; p = 0.001). In addition, the accuracy of combination of both indexes (94.6%; p = 0.04) could significantly improve as compared with DWI alone. CONCLUSION: DWI is more useful for the differentiation of SCLC from NSCLC than STIR, and their combination can significantly improve the accuracy in this setting. ADVANCES IN KNOWLEDGE: Pulmonary MRI, including DWI and STIR, had a potential of the suggestion of the possibility as SCLC.

Role of accumulated calcium in alleviating aluminum injury in wheat plants.

Aluminum (Al) sensitive wheat cultivar kalyansona was grown for 14 d in a range of Ca solution (125, 625, and 2500 µM) plus other nutrients without Al. At 14 d after Ca treatment, half of these plants were harvested (H1), and the rest of the plants were exposed to 100 µM Al for additional 6 d and harvested (H2). Severe Al injury was found only in the plants with the lowest supply of Ca before Al treatment. Aluminum concentration in the apoplastic fluid was very high at 125 µM Ca probably because the plasma membrane of some of the cells was destroyed due to the attack of 100 µM Al. Aluminum content in roots decreased with increasing supply of Ca before Al treatment. Calcium content decreased drastically at harvest (H2) in the plants with 100 µM Al. Under Al stress conditions, the plant responded to Al in different ways due to not only the different Ca supply but also the variation of Ca content in the plant tissues. Actually, the plants having the largest Ca content in the roots before Al treatment can receive less Al injury during Al treatment. To substantiate this idea, a companion study was conducted to investigate the effects of 2500 µM Ca supply during, before, and after 100 µM Al treatment on root growth. The results indicated clearly that exogenous Ca supply before Al treatment is able to alleviate Al injury but less effective than Ca supply during Al treatment.

Dobutamine stress echocardiography for assessment of systolic function in dogs with experimentally induced mitral regurgitation.

BACKGROUND: Systolic dysfunction is associated with poor outcomes in dogs with myxomatous mitral valve disease. However, assessment of systolic variables by conventional echocardiographic methods is difficult in these dogs because of mitral regurgitation (MR). HYPOTHESIS: We hypothesized that assessment of systolic function by dobutamine stress may identify systolic dysfunction in dogs with MR, and that 2-dimensional speckle-tracking echocardiography (2D-STE) could quantitatively evaluate myocardial function. ANIMALS: Anesthetized dogs with experimentally induced MR. METHODS: Dogs were examined for systolic myocardial deformations using 2D-STE during dobutamine infusion before and 3 and 6 months after MR induction. We evaluated peak systolic rotation and rotation rate in each basal and apical view; peak systolic torsion and torsion rate were also calculated. RESULTS: Invasive peak positive first derivatives of left ventricular pressure (dp/dt) were significantly decreased in dogs 6 months after induction of MR compared with pre-MR results. After 3 and 6 months of MR, dogs had diminished peak systolic torsion values and torsion rates in response to dobutamine infusion compared with pre-MR results (3 months, P < .001 and P = .006; 6 months, P = .003 and P = .021). These results were significantly correlated with overall invasive dp/dt (r = 0.644, P < .001; r = 0.696, P < .001). CONCLUSIONS AND CLINICAL IMPORTANCE: Decreased torsion during dobutamine infusion in dogs with MR may reflect latent systolic dysfunction. Dobutamine infusion, therefore, may be useful for the assessment of systolic function in dogs with MR.

Left ventricular geometrical differences in dogs with various stages of myxomatous mitral valve disease.

OBJECTIVES: To evaluate left ventricular geometry in dogs with various stages of myxomatous mitral valve disease. METHODS: Ninety-seven dogs with myxomatous mitral valve disease classified by the International Small Animal Cardiac Health Council system and 20 weight- and age-matched healthy dogs. Left ventricular long-axis to short-axis ratio, sphericity index in end-diastole and end-systole, left ventricular wall thickness to internal dimension ratio and relative wall thickness were assessed. RESULTS: The diastolic sphericity index was lower in classes Ib, II and III than in healthy dogs (P=0·003, P<0·001 and P<0·001) and was also lower in class III than in classes Ia, Ib and class II dogs (P<0·001, P<0·001 and P=0·002). The relative wall thickness was lower in classes II and III than in class Ia (P=0·003 and P<0·001), class Ib (P=0·004 and P<0·001), and healthy dogs (P<0·001 and P<0·001) and was also lower in class III than in class II (P=0·005). CLINICAL SIGNIFICANCE: Sphericity index and relative wall thickness are simple methods for assessing left ventricular geometry using two-dimensional echocardiography that may be useful in myxomatous mitral valve disease dogs as part of risk stratification.

Multiple inflammatory gastric polyps treated by endoscopic polypectomy with argon plasma coagulation in a dog.

An 11-year-old spayed female miniature dachshund was evaluated for a 2-month history of chronic vomiting. Abdominal ultrasonography revealed a heterogeneous mass in the pyloric region. Contrast upper gastrointestinal radiography demonstrated impairment of gastric outflow. Endoscopic examination revealed multiple polyps at the gastric pylorus. The pyloric polyps were variable in size, sessile-shaped and pedunculated. Initially, endoscopic polypectomy was attempted, but all the polyps could not be completely resected. Thus, endoscopic polypectomy with argon plasma coagulation was performed to cauterise the lesions. The histopathological diagnosis of the lesions was inflammatory polyps, and a moderate number of Helicobacter spp. was revealed. After the argon plasma coagulation treatment, the dog did not vomit, and improvement of clinical signs was maintained for 13 months. Endoscopic polypectomy with argon plasma coagulation may be useful for mixtures of sessile and pedunculated polyps. The present report may provide a basis for further studies of argon plasma coagulation treatment for canine gastrointestinal polyps.

Noninvasive clinical assessment of systolic torsional motions by two-dimensional speckle-tracking echocardiography in dogs with myxomatous mitral valve disease.

BACKGROUND: Left ventricular torsional motion plays an important role for effective pump function. However, noninvasive clinical assessment of torsional deformations by two-dimensional speckle-tracking echocardiography (2D-STE) in dogs with myxomatous mitral valve disease (MMVD) has not been reported. HYPOTHESIS: Left ventricular torsion is determined by the native orientation of the helical myocardial fibers, such that it might provide better assessment of myocardial function than conventional methods. ANIMALS: Sixty-seven client-owned dogs with MMVD were classified into 3 classes based on the International Small Animal Cardiac Health Council classification and 16 weight- and age-matched healthy dogs. METHODS: Dogs were examined for myocardial deformations by 2D-STE and were evaluated for peak systolic rotation and rotation rate at each basal and apical view. Dogs also were evaluated for peak systolic torsion and torsion rate. RESULTS: Peak systolic torsion was higher in class II than in class I (P < .001) dogs. Peak systolic torsion was lower in class III than in class II (P = .001) dogs and controls (P = .003). CONCLUSIONS AND CLINICAL IMPORTANCE: Torsional deformations assessed by 2D-STE differed among clinical classes of MMVD. Myocardial torsional deformations by 2D-STE may provide more detailed assessment of contractile function in dogs with MMVD.

Short communication: effects of serum obtained from dairy cows with low or high body condition score on in vitro embryo development.

The objective of the study was to determine whether the serum obtained from animals differing in body condition score (BCS) affects in vitro embryo development. After in vitro fertilization, serum obtained from dairy cows of either low (L-BCS; 2.1 ± 0.14 on a scale of 1 to 5) or high BCS (H-BCS; 4.0 ± 0.0), or commercially available bovine serum (control) was added at 5% concentration to the in vitro culture medium. Use of serum obtained from H-BCS cows increased the cleavage rates compared with control serum at both 24 and 48 h after in vitro fertilization (78.3 vs. 71.9% and 79.9 vs. 75.1%, respectively), whereas use of serum obtained from L-BCS cows increased the blastocyst rate compared with control serum at 7d (23.8 vs. 19.1%), but this difference was not evident at 8 or 9 d after in vitro fertilization. As nonesterified fatty acid concentrations were highest in control serum, followed by serum from L-BCS and H-BCS cows (621, 559, and 272 µEq/L, respectively), a high concentration of nonesterified fatty acids might adversely affect the very early stages of embryo development, and its negative effects might be greater immediately after fertilization compared with developmental stages after morula formation. Our findings also indicate that factors promoting early stage embryo development do not necessarily promote blastocyst development. Serum obtained from animals under different physiological conditions may be used for in vitro embryo culture to study the effects of nutritional management of dairy cattle on embryo development.

Three-dimensional airway lumen volumetry: comparison with bronchial wall area and parenchymal densitometry in assessment of airway obstruction in pulmonary emphysema.

OBJECTIVES: The purpose of this study was to compare three-dimensional airway lumen volumetry with bronchial wall area and parenchymal densitometry in the assessment of airway obstruction in pulmonary emphysema. METHODS: 56 patients, who were smokers, underwent CT examination and pulmonary function tests (PFTs). For quantitative assessments, the following parameters were computationally calculated: (1) percentage of voxels -950, -960 and -970 HU in the lung (%LAA(-950), %LAA(-960) and %LAA(-970), respectively); (2) percentage of partial bronchi luminal volumes per total luminal volumes (LV(main), main and distal bronchial volume/total luminal volume; LV(lobe), lobar and distal bronchial volume/total luminal volume); and (3) mean wall area percentages of segmental bronchi of the right apical and left apicoposterior segment (WA%(seg)) and of subsegmental bronchi (WA%(sub)) in the upper lobes. These parameters were correlated with PFTs and statistically compared between a chronic obstructive pulmonary disease (COPD) group [forced expiratory volume in 1 s (FEV1)/forced vital capacity (FVC)<70] and a non-COPD group. RESULTS: FEV1, maximum mid-expiratory flow rate and forced expiratory flow at 25% vital capacity had significant correlation with LV(main) (r>0.53, p<0.0001), LV(lobe) (r>0.52, p<0.0001), WA%(seg) (|r|>0.29, p<0.05) and WA%(sub) (|r|>0.31, p<0.05). FEV1/FVC had significant correlation with all parameters (0.27<|r|<0.52, p<0.05). LV(main) and LV(lobe) and WA%(seg) and WA%(sub) were significantly different between the two groups (LV(main) and LV(lobe); p<0.0001, WA%(seg) and WA%(sub); p<0.05). CONCLUSIONS: Bronchial luminal volumetric assessment better reflected the airflow limitation parameters. ADVANCES IN KNOWLEDGE: Bronchial luminal volumetric assessment can potentially be used to gauge airflow limitation in pulmonary emphysema.

Effects of folic acid on the development and oxidative stress of mouse embryos exposed to heat stress.

The development of mammalian pre-implantation embryos is inhibited by heat stress, and the inhibitory effect is associated with excess reactive oxygen species (ROS). Folate is a nutrient with various physiological functions including antioxidative effects. We first investigated the transcript expression for 10 enzymes in the cycle of folate metabolism (folate-methionine cycle) in mouse embryos at the 1-cell, 2-cell, 4- to 8-cell, morula and blastocyst stages using reverse transcription-polymerase chain reaction. All of the transcripts were consistently expressed, except for Mat1a, which was not detected from the 4- to 8-cell stage onward. Next, the effects of folic acid (the synthetic form of folate) on the development and ROS levels of heat-stressed embryos were investigated. One-cell mouse embryos were cultured with or without 1000 ng/ml folic acid basically at 38°C, and in the heat-stressed groups, embryos were exposed to 39.5°C/10 h/day on the first two days of culture. The heat stress significantly (p < 0.05) decreased blastocyst development and cell number and increased ROS levels compared to those in the group not subjected to heat stress; however, among the heat-stressed groups, blastocyst development and cell number were increased and the ROS level was decreased by the addition of folic acid. These results indicate that the mRNA of folate-methionine cycle enzymes are expressed in mouse pre-implantation embryos, suggesting they can independently utilize folate, and the inhibitory effects of heat stress on the development of mouse pre-implantation embryos are ameliorated by folic acid. The ameliorating effects of folic acid may be partly due to its antioxidative property.

Impact of polyplex micelles installed with cyclic RGD peptide as ligand on gene delivery to vascular lesions.

Gene therapy is expected to open a new strategy for the treatment of refractory vascular diseases, so the development of appropriate gene vectors for vascular lesions is needed. To realize this requirement with a non-viral approach, cyclo(RGDfK) peptide (cRGD) was introduced to block copolymer, poly(ethylene glycol)-block-polycation carrying ethylenediamine units (PEG-PAsp(DET)). cRGD recognizes α(v)ß(3) and α(v)ß(5) integrins, which are abundantly expressed in vascular lesions. cRGD-conjugated PEG-PAsp(DET) (cRGD-PEG-PAsp(DET)) formed polyplex micelles through complexation with plasmid DNA (pDNA) and the cRGD-PEG-PAsp(DET) micelles achieved significantly more efficient gene expression and cellular uptake as compared with PEG-PAsp(DET) micelles in endothelial cells and vascular smooth muscle cells. Intracellular tracking of pDNA showed that cRGD-PEG-PAsp(DET) micelles were internalized via caveolae-mediated endocytosis, which is associated with a pathway avoiding lysosomal degradation and that, PEG-PAsp(DET) micelles were transported to acidic endosomes and lysosomes via clathrin-mediated endocytosis. Further, in vivo evaluation in rat carotid artery with a neointimal lesion revealed that cRGD-PEG-PAsp(DET) micelles realized sustained gene expression, whereas PEG-PAsp(DET) micelles facilitated rapid, but transient gene expression. These findings suggest that introduction of cRGD to polyplex micelles might create novel and useful functions for gene transfer and contribute to the establishment of efficient gene therapy for vascular diseases.

Effects of bovine milk ingestion on urinary excretion of oxypurinol and uric acid.

OBJECTIVE: Although allopurinol is a xanthine oxidase inhibitor, its overall effect may be due to the action of oxypurinol, a metabolite of allopurinol and another xanthine oxidase inhibitor, since the biological half-life of oxypurinol is longer than that of allopurinol. Oxypurinol shares a renal transport pathway with uric acid and ingestion of bovine milk increases the urinary excretion of uric acid. Therefore, we investigated whether its ingestion promotes the urinary excretion of oxypurinol. SUBJECTS/METHODS: Bovine milk (15 ml/kg body weight) was administered to 6 healthy subjects who took allopurinol (300 mg) 12 h prior to ingestion. In addition, a control experiment was performed with the same subjects using the same protocol, except for the ingestion of water instead of bovine milk. Blood and urine samples were collected before and after bovine and water ingestion. RESULTS: In the bovine milk ingestion experiment, the urinary excretion values of oxypurinol and uric acid were increased by 18% and 38%, respectively, and the fractional excretion values of oxypurinol and uric acid were increased by 20% and 40%, respectively, whereas those did not change in the control experiment. In addition, the concentration of alanine and sum of concentrations of amino acids were increased by 16% and 20%, respectively, in the bovine milk ingestion experiment. CONCLUSION: These results suggest that bovine milk ingestion promotes the urinary excretion of oxypurinol as well as uric acid by increasing amino acid concentration.

Serum CRP in patients with gout and effects of benzbromarone.

OBJECTIVE: C-reactive protein (CRP) is associated with increased risk for myocardial infarction, atherosclerosis, and peripheral artery diseases, while increased serum uric acid level is suggested to be independently associated with an increased risk of cardiovascular mortality. Accordingly, to investigate whether hyperuricemia is associated with serum CRP, we compared serum CRP levels between healthy subjects and patients with gout. In addition, we also examined whether benzbromarone has effects on serum CRP levels in patients with gout and the expression of CRP messenger RNA of CRP in the hepatoma cell line HuH7. METHODS: In the first experiment, 40 healthy males and 43 male patients with gout were enrolled, then blood samples were drawn from each after an overnight fast. In the second experiment, 42 male patients with gout were given uric acid-lowering therapy with benzbromarone. Blood samples were drawn after an overnight fast before and 1 year after beginning benzbromarone treatment. In the third experiment, the effects of benzbromarone on IL1beta-induced CRP expression were determined in HuH7 cells. RESULTS: Log serum CRP levels were not significantly different between the patients with gout and healthy subjects, while log serum CRP levels were decreased by 11% after benzbromarone treatment, as compared to the values before treatment (p < 0.01). In addition, log serum adiponectin levels were elevated by 2% after treatment (p < 0.01). Furthermore, our in vitro findings demonstrated that benzbromarone down-regulated IL1beta-stimulated CRP gene expression. CONCLUSIONS: These results suggest that hyperuricemia may not contribute to an increase in serum CRP level, while benzbromarone may have a favorable effect on CRP.

Uracil-tegafur and tamoxifen vs cyclophosphamide, methotrexate, fluorouracil, and tamoxifen in post-operative adjuvant therapy for stage I, II, or IIIA lymph node-positive breast cancer: a comparative study.

BACKGROUND: It has been reported that treatment with uracil-tegafur (UFT) has shown significantly better survival and relapse-free survival (RFS) than surgery alone. Therefore, we compared UFT with a combination therapy of cyclophosphamide, methotrexate, and fluorouracil (CMF) in patients who had undergone curative surgery for axillary lymph node-positive breast cancer. METHODS: A total of 377 node-positive patients with stage I, II, or IIIA disease were registered from September 1996 through July 2000 and were randomly assigned to either 6 cycles of CMF or 2 years of UFT. In both arms, tamoxifen (TAM) was concurrently administered for 2 years. The primary end point in this study was the non-inferiority of UFT to CMF. RESULTS: No statistically significant difference between the two groups was observed with regard to the 5-year RFS rate (72.2% in the UFT and 76.3% in the CMF). Adverse event profiles differed between the two groups, with a significantly lower incidence of leukopenia and anaemia in the UFT group, as well as anorexia, nausea/vomiting, stomatitis, and alopecia, which have implications for quality of life. CONCLUSION: UFT administered in combination with TAM holds promise in the treatment of lymph node-positive early breast cancer. On stratified analysis, the recurrence rate in the UFT group was found to be better in oestrogen receptor (ER)-positive patients. Tegafur-based treatment should be evaluated by a prospective randomised trial conducted in ER-positive patients.