MicroRNA-383 located in frequently deleted chromosomal locus 8p22 regulates CD44 in prostate cancer.

A major genomic alteration in prostate cancer (PCa) is frequent loss of chromosome (chr) 8p with a common region of loss of heterozygosity (LOH) at chr8p22 locus. Genomic studies implicate this locus in the initiation of clinically significant PCa and with progression to metastatic disease. However, the genes within this region have not been fully characterized to date. Here we demonstrate for the first time that a microRNA component of this region-miR-383-is frequently downregulated in prostate cancer, has a critical role in determining tumor-initiating potential and is involved in prostate cancer metastasis via direct regulation of CD44, a ubiquitous marker of PCa tumor-initiating cells (TICs)/stem cells. Expression analyses of miR-383 in PCa clinical tissues established that low miR-383 expression is associated with poor prognosis. Functional data suggest that miR-383 regulates PCa tumor-initiating/stem-like cells via CD44 regulation. Ectopic expression of miR-383 inhibited tumor-initiating capacity of CD44+ PCa cells. Also, 'anti-metastatic' effects of ectopic miR-383 expression were observed in a PCa experimental metastasis model. In view of our results, we propose that frequent loss of miR-383 at chr8p22 region leads to tumor initiation and prostate cancer metastasis. Thus, we have identified a novel finding that associates a long observed genomic alteration to PCa stemness and metastasis. Our data suggest that restoration of miR-383 expression may be an effective therapeutic modality against PCa. Importantly, we identified miR-383 as a novel PCa tissue diagnostic biomarker with a potential that outperforms that of serum PSA.

Engineering a Spin-Orbital Magnetic Insulator by Tailoring Superlattices.

In 5d Ir oxides with an interplay of spin-orbit coupling and electron correlations, we have tailored a spin-orbital magnetic insulator out of a semimetal SrIrO(3) by tuning the structure through superlattices [(SrIrO(3))(m), SrTiO(3)] (m=1,2,3,4, and ∞). We observed the systematic decrease of the magnetic ordering temperature and the resistivity as a function of m. The transition from the semimetal to the insulator is found to be closely linked to the appearance of magnetism at m≃3. Long range magnetic ordering was realized even in the m=1 single layer superlattice, implying that the design and realization of novel electronic phases is feasible at the level of a single atomic layer in complex Ir oxides.

Increased phosphatidylcholine concentration in saliva reduces surface tension and improves airway patency in obstructive sleep apnoea.

Surface tension may have important role for maintaining upper airway patency in patients with obstructive sleep apnoea. It has been demonstrated that elevated surface tension increases the pharyngeal pressures required to reopen the upper airway following collapse. The aim of the study was to evaluate the associations between the concentrations of endogenous surfactants in saliva with indices of upper airway patency in obstructive sleep apnoea. We studied 20 male patients with obstructive sleep apnoea (age: 60·3 ± 10·3 years; BMI: 25·9 ± 4·6 kg m(-2); AHI: 41·5 ± 18·6 events h(-1)). We obtained 100-µL samples of saliva prior to overnight polysomnographic sleep study. The surface tension was determined using the pull-off force technique. The concentration of phosphatidylcholine (PC) was evaluated by liquid chromatography-mass spectrometry (LC-MS/MS). Regression analysis between apnoea, hypopnoea and apnoea/hypopnoea indices and the ratio of hypopnoea time/total disordered breathing time (HT/DBT) with surface tension and PC were performed. P < 0·05 was considered significant. The mean saliva surface tension was 48·8 ± 8·0 mN m(-1) and PC concentration was 15·7 ± 11·1 nM. The surface tension was negatively correlated with the PC concentration (r = -0·48, P = 0·03). There was a significant positive correlation between surface tension with hypopnoea index (r = 0·50, P = 0·03) and HT/DBT (r = 0·6, P = 0·006), but not apnoea or apnoea/hypopnoea index (P > 0·11). Similarly, PC concentration negatively correlated with hypopnoea index (r = -0·45, P = 0·04) and HT/DBT (r = -0·6, P = 0·004), but not with apnoea index or AHI (P > 0·08). An increase in salivary PC concentration may increase upper airway patency in obstructive sleep apnoea through a reduction in surface tension.

Protonation behavior and solution properties of amine oxide surfactants containing a pyridyl group.

Hydrogen bonding between surfactant molecules plays an important role in self-assembly formation. For long alkyl chain amine oxide surfactants, the specific protonation degree dependence of some solution properties has been considered to be due to hydrogen bonding between protonated and deprotonated species. In addition to this type of hydrogen bonding, we introduced a pyridyl group into an alkylamine oxide molecule as a new hydrogen-bonding site. The pyridyl group has three different structural isomers based on the position of the substituent. An amine oxide group in pyridylamine oxides was preferentially protonated. In addition, protonation of the pyridyl group revealed a pronounced substituent position effect on the critical micelle concentration, micellar size, and solubilization of oil-soluble dye into micelles. The intermolecular or intramolecular hydrogen bond formation could be controlled by altering the substituent position.

Reduction in radiation and contrast medium dose via optimization of low-kilovoltage CT protocols using a hybrid iterative reconstruction algorithm at 256-slice body CT: phantom study and clinical correlation.

AIM: To optimize low-kilovoltage (kV) computed tomography (CT) protocols using a hybrid iterative reconstruction (HIR) algorithm at 256-detector-row body CT. MATERIALS AND METHODS: Based on preliminary phantom studies, three different tube voltage protocols with an equal contrast-to-noise ratio (CNR) were developed. They were a conventional 120 kV protocol with filtered back-projection (FBP), an 80 kV protocol with HIR (a 160% increase in the tube current-time product and a 40% reduction in the contrast medium dose), and a 100 kV protocol with HIR (a 20% reduction in the tube current-time product and the contrast medium dose). The clinical study included 70 patients (34 women, 36 men; mean age 70.5 ± 9.1 years, range 44-92 years) who had undergone CT at 120 kV a mean of 148 ± 137 days before undergoing low kV contrast-enhanced body CT (80 kV with HIR, n = 35; 100 kV with HIR, n = 35). The estimated effective radiation dose (ED), image noise, and CNR were calculated and the visual image quality was scored on a four-point scale. RESULTS: Mean ED was 12.3, 8.4, and 15.4 mSv for the 80, 100, and 120 kV protocol, respectively, and significantly lower using the low kV protocols. There was no significant difference in the image noise and CNR between the low kV protocols with HIR and the 120 kV protocol with FBP, or in the visual scores among the three protocols. CONCLUSION: Without ensuing image-quality degradation, the radiation and contrast medium dose can be reduced with optimal contrast-enhanced CT protocols using a low kV technique and an HIR algorithm.

Zonisamide enhances delta receptor-associated neurotransmitter release in striato-pallidal pathway.

A recent randomized control study demonstrated that zonisamide (ZNS), an antiepileptic drug, is effective in Parkinson's disease at the lower than the therapeutic doses against epilepsy (25-50 mg/day); however, the detailed mechanism of antiparkinsonian effects of ZNS remains to be clarified. To determine the mechanism of antiparkinsonian effect of ZNS, we investigated the effects of ZNS on extracellular levels of dopamine in the striatum (STR), glutamate in substantia nigra pars reticulata (SNr), GABA in globus pallidus (GP), subthalamic nucleus (STN) and SNr, using multiple microdialysis probes. Striatal perfusion of 1000 microM ZNS (within therapeutic-relevant concentration against epilepsy) increased extracellular levels of dopamine in STR, whereas 100 microM ZNS (lower than the therapeutic-relevant concentration against epilepsy but within the therapeutic rage against Parkinson's disease) did not affect it. Striatal perfusion of ZNS (100 and 1000 microM) decreased the extracellular levels of GABA in STN and glutamate in SNr, but decreased extracellular GABA level in GP without affecting GABA level in SNr. These concentration-dependent effects of ZNS on extracellular neurotransmitter levels were independent of dopamine and delta(2) receptors; however, blockade of delta(1) receptor inhibited the effects of ZNS. Furthermore, activation of delta(1) receptor enhanced the effects of ZNS on neurotransmitter level. These results suggest that ZNS does not affect the direct pathway but inhibits the indirect pathway, which is mediated by delta(1) receptor. Therefore, the antiparkinsonian effects of ZNS seem to be mediated through the interaction between lower than therapeutically-relevant concentration against epilepsy of ZNS (100 microM) and delta(1) receptor.

Effects of zotepine on extracellular levels of monoamine, GABA and glutamate in rat prefrontal cortex.

BACKGROUND AND PURPOSE: The atypical antipsychotic drug, zotepine, is effective in treatment of schizophrenia and acute mania, but the incidence of seizures during treatment is higher than with other antipsychotics. In addition, the mechanisms underlying the clinical actions of zotepine remain uncharacterized. EXPERIMENTAL APPROACH: The effects of intraperitoneal administration of zotepine and haloperidol on the extracellular levels of noradrenaline, dopamine, 5-HT, GABA, and glutamate in the medial prefrontal cortex (mPFC) were compared. Neuronal activities induced by each drug in the ventral tegmental area (VTA), locus coeruleus (LC), dorsal raphe nucleus (DRN) and mediodorsal thalamic nucleus (MTN) were also analysed. KEY RESULTS: Haloperidol did not affect extracellular neurotransmitter levels in the mPFC. In contrast, zotepine activated neuronal activities in all nuclei and increased the extracellular levels of noradrenaline, dopamine, GABA, and glutamate in the mPFC, but not 5-HT levels. The zotepine-stimulated neuronal activity in the VTA, LC, DRN and MTN enhanced the release of dopamine, noradrenaline, 5-HT, glutamate and GABA in the mPFC, although the enhanced GABAergic transmission possibly inhibited noradrenaline, dopamine and 5-HT release. The other afferent to mPFC, which releases dopamine and noradrenaline, was partially insensitive to GABAergic inhibition, but possibly received stimulatory AMPA/glutamatergic regulation from the MTN. CONCLUSIONS AND IMPLICATIONS: Our results indicated that the positive interaction between prefrontal catecholaminergic transmission and AMPA/glutamatergic transmission from MTN might explain the regulatory effects of zotepine on neurotransmitter release. A mechanism is suggested to account for the pharmacological profile of this atypical antipsychotic and for its pro-convulsive action.

The effect of Lactobacillus helveticus fermented milk on sleep and health perception in elderly subjects.

OBJECTIVE: To study the effect of Lactobacillus helveticus fermented milk on sleep and health perception in elderly healthy subjects. SUBJECTS: The study included 29 healthy elderly subjects aged 60-81 years. METHODS: Prospective, randomized, double-blind and placebo-controlled, with a crossover design. The study included two intervention periods of 3 weeks each, separated by a 3-week washout period. Subjects took 100 g of fermented milk drink or a placebo drink (artificially acidified milk) daily in the first supplementary period and the other drink in the second supplementary period. For each period, we measured sleep quality by means of actigraphy and a sleep questionnaire, and assessed the quality of life (QOL) by SF-36 health survey. RESULTS: There was a significant improvement in sleep efficiency (P=0.03) and number of wakening episodes (P=0.007) in actigraph data after intake of fermented milk, whereas no significant changes were observed for the placebo. Fermented milk did not improve the SF-36 scores significantly from the baseline period. In the GH domain (general health perception) of the SF-36, however, there was marginal improvement as compared to the baseline period. Although the difference between fermented milk and placebo was not statistically significant for any of the sleep or QOL parameters, fermented milk produced slightly greater mean values for many parameters. CONCLUSION: This short-term (3-week) intervention study indicates that Lactobacillus helveticus fermented milk may have a more favorable effect on improving sleep in healthy elderly people as compared with placebo.

Application of artificial neural network modelling to identify severely ill patients whose aminoglycoside concentrations are likely to fall below therapeutic concentrations.

OBJECTIVE: Identification of ICU patients whose concentrations are likely to fall below therapeutic concentrations using artificial neural network (ANN) modelling and individual patient physiologic data. METHOD: Data on indicators of disease severity and some physiologic data were collected from 89 ICU patients who received arbekacin (ABK) and 61 who received amikacin (AMK). Three-layer ANN modelling and multivariate logistic regression analysis were used to predict the plasma concentrations of the aminoglycosides (ABK and AMK) in the severely ill patients. RESULTS: Predictive performance analysis showed that the sensitivity and specificity of ANN modelling was superior to multivariate logistic regression analysis. For accurate modelling, a predictable range should be inferred from the data structure before the analysis. Restriction of the predictable region, based on the data structure, increased predictive performance. CONCLUSION: ANN analysis was superior to multivariate logistic regression analysis in predicting which patients would have plasma concentrations lower than the minimum therapeutic concentration. To improve predictive performance, the predictable range should be inferred from the data structure before prediction. When applying ANN modelling in clinical settings, the predictive performance and predictable region should be investigated in detail to avoid the risk of harm to severely ill patients.

Preparation and characterization of glassy celecoxib.

Celecoxib, a poorly water-soluble drug, was converted into a glassy state by melt quenching. The properties of glassy celecoxib were studied using infrared (IR) spectroscopy, differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), intrinsic dissolution rate (IDR), and thin-layer-chromatography (TLC). Glass transition occurred at 51.8 degrees C. Infrared spectrum of glass has revealed significant changes due to H-bonding. Celecoxib glass shows around 15 times faster dissolution as compared with the crystalline state. Heckel plot analysis has shown better compressibility in glassy state. Unpulverized glass remained stable for 3 months, whereas after pulverization about 70% crystallinity was gained in 100 hours. Further attempts may be carried out to stabilize the glass.

Remodelling and healing process of moderately heat-treated bone grafts after wide resection of bone and soft-tissue tumors.

INTRODUCTION: In the reconstruction of extensive bone defects after massive resection of malignant musculoskeletal tumors, the clinical results of moderately heat-treated autogenous bone graft have rarely been documented. We evaluated the remodelling and healing process of moderately heat-treated autogenous bone graft by means of imaging features. MATERIALS AND METHODS: The subjects of this study were 19 patients with bone and soft-tissue tumors treated by heat-treated bone graft at our institution between 1992 and 2001, the mean follow-up period was 4.8+/-2.8 years (range 1-9 years). The remodelling and healing process of heat-treated bone graft was evaluated by means of radiography, bone scintigraphy, and MRI. RESULTS: The mean period to obtain bone union between host bone and grafted bone was 9.4 months. Infection was noted in 1 patient, and fracture was present in 2 patients. In 6 patients, pseudoarthrosis was found. Bone scintigraphy showed an increased uptake at the host-graft junction in the period between 3 and 36 months (median 10.7 months) postoperatively. A gradually increased diffuse uptake on the grafted side was evident at an average of 29.1 months (range 19-41 months) postoperatively. High signal intensity on T2-weighted images was observed in the early period after surgery, and iso-intense or low signal intensity became evident after an average of 28.3 months. A gradually increased diffuse uptake on scintigraphy and iso-intense or low signal intensity on T2-weighted images indicated remodelling of the grafted bone. CONCLUSION: Bone union of a moderately heat-treated autogenous bone graft was noted at about 9 months, and its remodelling was proceeding at about 30 months. This method will be useful for bone defects after massive resection of soft-tissue and bone tumors.

Myxoid liposarcoma metastatic to the thoracic epidural space without bone involvement: report of two cases.

Myxoid liposarcoma can frequently metastasize to extrapulmonary sites. We present two cases of myxoid liposarcoma metastatic to the epidural space. Both patients complained of back pain, but plain radiography revealed no abnormality. MR imaging clearly demonstrated metastatic tumors in the epidural space, but no involvement of vertebra. When patients with myxoid liposarcoma complain of back pain, metastasis in the epidural space should be considered even in patients without bone involvement.

[Treatment outcomes with vinorelbine for metastatic breast cancer patients previously treated with both doxorubicin and docetaxel].

The purpose of this study was to evaluate the efficacy and toxicity of weekly vinorelbine (VNB) in patients with metastatic breast cancer previously treated with both adriamycin (ADM) and docetaxel (TXT). VNB was administered weekly at the dose 20 mg/m2 by i.v. infusion over 20 minutes followed by flushing the vein with 100 ml of normal saline. From June 1999 to August 2000, ten patients were enrolled in this study. Patient characteristics were that the cumulative doses (median) of previous ADM and TXT were 300 mg (range, 120-880 mg), 560 mg (range, 120-960 mg) respectively. The median number of metastatic sites was four, with poor performance status (ECOG 1-2: 40%, 3-4: 60%). The median cycles of weekly VNB were seven (range: 2-12). Two of 10 assessable patients obtained partial response, with an overall response rate of 20%. The main toxicity (NCI grade 4) was leukopenia in 10% of 10 patients. Phlebitis (grade 2) was observed in 4 of 10 patients (40%). VNB is an active agent against metastatic breast cancer pretreated with both ADM and TXT, possessing no severe toxicities.

Differential effects of protein kinase C on human vascular smooth muscle cell proliferation and migration.

Vascular smooth muscle cell (SMC) migration and proliferation contribute to intimal hyperplasia, and protein kinase C (PKC) may be required for both events. In this report, we investigated the role of PKC in proliferation and migration of SMC derived from the human saphenous vein. Activation of PKC by phorbol-12,13-dibutyrate (PDBu) or (-)-indolactam [(-)-ILV] increases SMC proliferation. Downregulation of PKC activity by prolonged incubation with phorbol ester or inhibition of PKC with chelerythrine in SMC diminished agonist-stimulated proliferation. In contrast, stimulation of PKC with PDBu or (-)-ILV inhibited basal and agonist-induced SMC chemotaxis. Moreover, downregulation of PKC or inhibition with chelerythrine accentuated migration. We postulated that the inhibitory effect of PKC on SMC chemotaxis was mediated through cAMP-dependent protein kinase (protein kinase A, PKA). In support of this hypothesis, we found that activation of PKC in SMC stimulated PKA activity. The cAMP agonist forskolin significantly inhibited SMC chemotaxis. Furthermore, the inhibitory effect of PKC on SMC chemotaxis was completely reversed by cAMP or PKA inhibitors. In search of the PKC isotype(s) underlying these differential effects of PKC in SMC, we identified eight isotypes expressed in human SMC. Only PKC-alpha, -beta I, -delta, and -epsilon were eliminated by downregulation, suggesting that one or more of these four enzymes facilitate the observed phorbol ester-dependent effects of PKC in SMC. In summary, we found that PKC activation enhances proliferation but inhibits migration of human vascular SMC. These differential effect of PKC on vascular cells appears to be mediated through PKC-alpha, -beta I, -delta, and/or -epsilon.

The change in characteristics of microcrystalline cellulose during wet granulation using a high-shear mixer.

The objective of this study was to investigate the mechanism of hard granule formation and to demonstrate the applicability of X-ray diffraction methods for studying the polymeric pharmaceutical excipients. Using a high-shear mixer, microcrystalline cellulose (MCC) was granulated with water as the granulating liquid. The hardness of the MCC granules increased with granulation time and the amount of water added. The specific surface area measured by the N2 adsorption method was reduced during the process. Crystallite size of cellulose, calculated by Scherrer's equation adapted for wide angle X-ray diffraction method, decreased with granulation time and with increasing amounts of water added. Debye plots for X-ray small scattering patterns suggested that the average magnitude of the continuous solid region in MCC granules became significantly greater, whereas the specific surface area of the MCC granules, calculated from Debye plots, became smaller in comparison with that of intact MCC. These findings suggested that the long-chain structures in MCC were disrupted, resulting in smaller units with shorter chain lengths due to the strong shear force of the impeller. These smaller units then form a network within the granules. Thus, MCC granules are strengthened with longer granulation time and greater amounts of water, resulting in a more intricate network. The change in MCC chain length and physical structure can be experimentally detected using the small-angle X-ray scattering and wide-angle powder X-ray diffraction methods.

A species-selective allelopathic substance from germinating sunflower (Helianthus annuus L.) seeds.

From the exudate of germinating sunflower (Helianthus annuus L.) seeds was isolated a stereoisomer of diversifolide, 4, 15-dinor-3-hydroxy-1(5)-xanthene-12,8-olide (designated sundiversifolide) as determined by analysis of its IR, APCI-, ESI- and HR-MS and 13C and 1H NMR spectra. This substance inhibited shoot and root growth of cat's-eyes by about 50% at a concentration of 30 ppm. It also showed species-selective activity on the shoot and root growth of tested plants. When cat's-eyes seeds were incubated together with sunflower seeds, the cat's-eyes growth was inhibited. Furthermore, it was detected from an extract of river sand when sunflower seeds were incubated on the sand. These results indicate that sundiversifolide has an allelopathic function in sunflower plants.

Walking ability and activities of daily living after limb salvage operations for malignant bone and soft-tissue tumors of the lower limbs.

We evaluated the number of steps, activities of daily life (ADL) score, Enneking score, active range of motion and muscle strength by muscle manual testing for function in lower limbs after reconstructive procedures in surgical treatment of tumors. The 56 patients with 20 malignant bone tumors and 36 malignant soft-tissue tumors averaged 7119 +/- 3563 steps per day, or 69.8% of the control group. The average ADL score of patients was 14.0 +/- 4.1 points (70.0%), and the average Enneking score 20.4 +/- 6.0 points (68.0%). The scores of the bone tumor group were lower than those of the soft-tissue tumor group. These scores were not correlated with the range of motion. The number of steps and ADL score were correlated with Enneking score (coefficient 0.52 and 0.84, respectively). The number of steps and the ADL score appear to be useful, as is Enneking score.

Quantitative analysis of crystalline pharmaceuticals in powders and tablets by a pattern-fitting procedure using X-ray powder diffraction data.

A pattern-fitting procedure for quantitative analysis of crystalline pharmaceuticals in solid dosage forms using X-ray powder diffraction data is described. This method is based on a procedure for pattern-fitting in crystal structure refinement, and observed X-ray scattering intensities were fitted to analytical expressions including some fitting parameters, i.e. scale factor, peak positions, peak widths and degree of preferred orientation of the crystallites. All fitting parameters were optimized by the non-linear least-squares procedure. Then the weight fraction of each component was determined from the optimized scale factors. In the present study, well-crystallized binary systems, zinc oxide-zinc sulfide (ZnO-ZnS) and salicylic acid-benzoic acid (SA-BA), were used as the samples. In analysis of the ZnO-ZnS system, the weight fraction of ZnO or ZnS could be determined quantitatively in the range of 5-95% in the case of both powders and tablets. In analysis of the SA-BA systems, the weight fraction of SA or BA could be determined quantitatively in the range of 20-80% in the case of both powders and tablets. Quantitative analysis applying this pattern-fitting procedure showed better reproducibility than other X-ray methods based on the linear or integral intensities of particular diffraction peaks. Analysis using this pattern-fitting procedure also has the advantage that the preferred orientation of the crystallites in solid dosage forms can be also determined in the course of quantitative analysis.