Angiopoietin-like protein 2 promotes chondrogenic differentiation during bone growth as a cartilage matrix factor.

OBJECTIVE: Chondrocyte differentiation is crucial for long bone growth. Many cartilage extracellular matrix (ECM) proteins reportedly contribute to chondrocyte differentiation, indicating that mechanisms underlying chondrocyte differentiation are likely more complex than previously appreciated. Angiopoietin-like protein 2 (ANGPTL2) is a secreted factor normally abundantly produced in mesenchymal lineage cells such as adipocytes and fibroblasts, but its loss contributes to the pathogenesis of lifestyle- or aging-related diseases. However, the function of ANGPTL2 in chondrocytes, which are also differentiated from mesenchymal stem cells, remains unclear. Here, we investigate whether ANGPTL2 is expressed in or functions in chondrocytes. METHODS: First, we evaluated Angptl2 expression during chondrocyte differentiation using chondrogenic ATDC5 cells and wild-type epiphyseal cartilage of newborn mice. We next assessed ANGPTL2 function in chondrogenic differentiation and associated signaling using Angptl2 knockdown ATDC5 cells and Angptl2 knockout mice. RESULTS: ANGPTL2 is expressed in chondrocytes, particularly those located in resting and proliferative zones, and accumulates in ECM surrounding chondrocytes. Interestingly, long bone growth was retarded in Angptl2 knockout mice from neonatal to adult stages via attenuation of chondrocyte differentiation. Both in vivo and in vitro experiments show that changes in ANGPTL2 expression can also alter p38 mitogen-activated protein kinase (MAPK) activity mediated by integrin α5ß1. CONCLUSION: ANGPTL2 contributes to chondrocyte differentiation and subsequent endochondral ossification through α5ß1 integrin and p38 MAPK signaling during bone growth. Our findings provide insight into molecular mechanisms governing communication between chondrocytes and surrounding ECM components in bone growth activities.

Nanodiamond finding in the hyblean shallow mantle xenoliths.

Most of Earth's diamonds are connected with deep-seated mantle rocks; however, in recent years, µm-sized diamonds have been found in shallower metamorphic rocks, and the process of shallow-seated diamond formation has become a hotly debated topic. Nanodiamonds occur mainly in chondrite meteorites associated with organic matter and water. They can be synthesized in the stability field of graphite from organic compounds under hydrothermal conditions. Similar physicochemical conditions occur in serpentinite-hosted hydrothermal systems. Herein, we report the first finding of nanodiamonds, primarily of 6 and 10 nm, in Hyblean asphaltene-bearing serpentinite xenoliths (Sicily, Italy). The discovery was made by electron microscopy observations coupled with Raman spectroscopy analyses. The finding reveals new aspects of carbon speciation and diamond formation in shallow crustal settings. Nanodiamonds can grow during the hydrothermal alteration of ultramafic rocks, as well as during the lithogenesis of sediments bearing organic matter.

Mechanisms inducing autonomic dysreflexia during urinary bladder distention in rats with spinal cord injury.

OBJECTIVES: This study investigated the mechanisms inducing autonomic dysreflexia due to enhanced bladder-to-vascular reflexes in rats with spinal cord injury (SCI). METHODS: SCI was produced by the transection of the Th4-5 spinal cord in female Sprague-Dawley rats. At 4 weeks after SCI, changes in blood pressure during graded increases in intravesical pressure (20-60 cm H2O) were measured in spinal-intact (SI) and SCI rats under urethane anesthesia. In five animals, effects of C-fiber desensitization induced by intravesical application of resiniferatoxin (RTX), a TRPV1 agonist, on the bladder-to-vascular reflex were also examined. Nerve growth factor (NGF) levels of mucosa and detrusor muscle layers of the bladder were measured by enzyme-linked immunosorbent assay. The expression levels of TRPV1 and TRPA1 channels were also examined in laser captured bladder afferent neurons obtained from L6 DRG, which were labeled by DiI injected into the bladder wall. RESULTS: In SI and SCI rats, systemic arterial blood pressure was increased in a pressure-dependent manner during increases in the intravesical pressure, with significantly higher blood pressure elevation at the intravesical pressure of 20 cm H2O in SCI rats vs SI rats. The arterial blood pressure responses to bladder distention were significantly reduced by RTX-induced desensitization of C-fiber bladder afferent pathways. SCI rats had higher NGF protein levels in the bladder and higher TRPV1 and TRPA1 mRNA levels in bladder afferent neurons compared with SI rats. CONCLUSIONS: The bladder-to-vascular reflex induced by TRPV1-expressing C-fiber afferents during bladder distention is enhanced after SCI in association with increased expression of NGF in the bladder and TRP channels in bladder afferent neurons.

Simultaneous modified Camitz opponensplasty using a pulley at the radial side of the flexor retinaculum in severe carpal tunnel syndrome.

Camitz opponensplasty using the palmaris longus has been used in patients undergoing open carpal tunnel release. It is considered to have several advantages over other opponensplasty techniques, but it provides weak flexion and pronation, which are prerequisites for opposition. To address this shortcoming, we have used a modified Camitz procedure with a pulley at the radial side of the dissected flexor retinaculum and have assessed the results in comparison with the conventional Camitz procedure. Both procedures provided significant improvements in palmar abduction and Disabilities of the Arm, Shoulder, and Hand and Kapandji scores at 3 months post-operatively, but patients who underwent the modified Camitz procedure showed better improvement in pulp pinch, palmar abduction, and thumb pronation.

Retroperitoneal approach for laparoscopic nephroureterectomy with stripping technique: extracorporeal ligation of ureter and ureteral catheter.

INTRODUCTION: The pluck and stripping techniques are used for lower ureter management in renal pelvic cancer patients. Herein, we report our experience of extracorporeal ligation of the ureter and the ureteral catheter through the trocar port, which differs from conventional laparoscopic ligation in the retroperitoneal space. This technique was selected to reduce the time needed for ureter management using the stripping technique and to provide secure ligation. MATERIALS AND SURGICAL TECHNIQUE: We performed this stripping technique in patients with T1 and T2 stage renal pelvic cancer without imaging-evident lymph node metastasis. After transurethrally placing a ureteral catheter, we resected the circumference of the ureteral orifice. After laparoscopic nephrectomy via a retroperitoneal approach, the ureteral catheter and distal ureter were ligated extracorporeally. The catheter was pulled to invaginate the ureter so it could then be pulled through the external urethral orifice. DISCUSSION: This technique of extracorporeal ligation ensures more a secure ligation of the ureter and ureteral catheter. This modified stripping technique does not require lower ureter management with laparotomy, and it is also useful in shortening the operative time. This method is effective for relatively early stage renal pelvic cancer.

Phase I study of paclitaxel, cisplatin and 5-fluorouracil combination chemotherapy for unresectable / recurrent gastric cancer.

PURPOSE: We investigated the safety of triple combination therapy by addition of Paclitaxel (PTX) to Cisplatin (CDDP) and 5-fluorouracil (5-FU) combination therapy, which was considered the conventional standard therapy for patients with unresectable / recurrent gastric cancer. MATERIAL AND METHODS: The doses of PTX and CDDP were fixed at 80 and 50 mg/m2. They were administered on days 1 and 8, followed by a resting period of 20 days. 5-FU 300 mg/m2 at a maximum dose of 500 mg/m2 was administered at levels 0 and 2, respectively, and the dose was increased by 100 mg/m2 until the maximum tolerated dose (MTD). It was administered on days 1 - 5 and 8 - 12, followed by a resting period of 16 days. RESULTS: Twelve patients enrolled in this study. Of them, three patients were excluded from evaluation because treatment continuation was not feasible. There were 4 leukopenia and 7 neutropenia cases with hematological toxicity at grade 3 or higher. They were observed at all dose levels, but no case showed infection. In terms of non-hematological toxicity at grade 3 or higher, there were two patients with nausea and vomiting and two patients with diarrhea, one patient with mucositis, one patient with anorexia. All patients with non-hematological toxicity at grade 3 or higher were at level 2. The dose-limiting toxicity (DLT) was observed at level 2, and 5-FU at 400 mg (level 1) was adopted. CONCLUSIONS: We proved in this study that PTX, CDDP, and 5-FU combination chemotherapy was a safe treatment.

Transvaginal mesh (TVM) reconstruction with TVT/TOT sling for vaginal prolapse concurrent with stress urinary incontinence.

We evaluated clinical efficacies of transvaginal mesh (TVM) reconstruction alone and those concomitant with a TVT/TOT sling for the treatment of pelvic organ prolapse (POP) and stress urinary incontinence (SUI). Between January 2006 and February 2007, 138 female patients with POP underwent TVM reconstruction. The mean age was 66.6 years (range: 52-84). Fourteen individuals were qualified as grade II in the POP quantification (POP-Q) system, 85 and 39 were grades III and IV, respectively. One hundred and seventeen of 138 (85 %) cases showed SUI. Twenty-one patients without SUI underwent TVM alone, and 117 cases with SUI underwent TVM concomitant with TVT/TOT sling. Mean operation time and intra-operative bleeding was 79 min (range: 25-177) and 74 ml (range: 10-429), respectively. Mean follow-up period is 5.3 months (range: 1-14). The vaginal prolapses were cured (grade 0) in 129 cases (93 %) after the surgery. Total inter-national prostate symptom score (IPSS), its QOL score, International Consultation on Incontinence Questionnaires Short Form (ICIQ-SF) significantly improved (from 12.6 to 3.9; p < 0.0001, from 5.0 to 1.0; p < 0.0001, and from 6.1 to 2.5; p < 0.01, respectively). Six of 21 cases (29%) who underwent TVM alone showed postoperative de-novo SUI. In contrast, 116 cases (99%) who underwent TVM concomitant with TVT/TOT, experienced a cure of SUI. Maximum flow rate did not change postoperatively in the both groups. In conclusion, the short-term efficacies of TVM reconstruction for POP are excellent, and a concomitant TVT/TOT sling prevents postoperative SUI.

T-type calcium channel blockade as a therapeutic strategy against renal injury in rats with subtotal nephrectomy.

T-type calcium channel blockers have been previously shown to protect glomeruli from hypertension by regulating renal arteriolar tone. To examine whether blockade of these channels has a role in protection against tubulointerstitial damage, we used a stereo-selective T-type calcium channel blocker R(-)-efonidipine and studied its effect on the progression of this type of renal injury in spontaneously hypertensive rats that had undergone subtotal nephrectomy. Treatment with racemic efonidipine for 7 weeks significantly reduced systolic blood pressure and proteinuria. The R(-)-enantiomer, however, had no effect on blood pressure but significantly reduced proteinuria compared to vehicle-treated rats. Both agents blunted the increase in tubulointerstitial fibrosis, renal expression of alpha-smooth muscle actin and vimentin along with transforming growth factor-beta (TGF-beta)-induced renal Rho-kinase activity seen in the control group. Subtotal nephrectomy enhanced renal T-type calcium channel alpha1G subunit expression mimicked in angiotensin II-stimulated mesangial cells or TGF-beta-stimulated proximal tubular cells. Our study shows that T-type calcium channel blockade has renal protective actions that depend not only on hemodynamic effects but also pertain to Rho-kinase activity, tubulointerstitial fibrosis, and epithelial-mesenchymal transitions.

A novel macrocyclic spermine alkaloid from Incarvillea sinensis.

A novel macrocyclic spermine alkaloid incasine C' (1), along with a known compound incasine C (2), were isolated from the whole plants of Incarvillea sinensis, and their structures were elucidated on the basis of chemical and spectroscopic evidence.

Evaluation of a method to determine the natural occurrence of aflatoxins in commercial traditional herbal medicines from Malaysia and Indonesia.

Traditional herbal medicines, popularly known as 'jamu' and 'makjun' in Malaysia and Indonesia, are consumed regularly to promote health. In consideration of their frequent and prolonged consumption, the natural occurrence of aflatoxins (AF) in these products was determined using immunoaffinity column clean-up and high-performance liquid chromatography with pre-column derivatization. The evaluated method, which entails dilution of sample extracts with Tween 20-phosphate buffered saline (1:9, v/v) and a chromatographic system using isocratic mobile phase composed of water-methanol-acetonitrile (70:20:10, v/v/v), was effective in separating AFB1, AFG1 and AFG2 from interference at their retention times. Results were confirmed using post-column derivatization with photochemical reactor. For 23 commercial samples analyzed, mean levels (incidence) of AFB(1), AFB(2) and AFG1 in positive samples were 0.26 (70%), 0.07 (61%) and 0.10 (30%) microg/kg, respectively; one sample was positive for AFG2 at a level of 0.03 (4%) microg/kg. In contrast to the high levels of AF in crude herbal drugs and medicinal plants reported previously by other researchers, the low contamination levels reported in this study may be attributed to the higher selectivity to AF of the method applied. Based on the AFB1 levels and the daily consumption of positive samples, a mean probable daily intake of 0.022 ng/kg body weight was calculated.

Microbiological and physicochemical factors affecting Aspergillus section Flavi incidence in Cavendish banana (Musa cavendishii) chips production in Southern Philippines.

Microbiological and physicochemical factors affecting the incidence of Aspergillus section Flavi in dried Cavendish banana (Musa cavendishii) chips production in Southern Philippines were examined. The average counts of Aspergillus section Flavi (AFC) in fresh and dried Cavendish bananas from 10 production batches of the Philippine Agro-Industrial Development Cooperative in Davao del Norte, Southern Philippines were 1.2 x 10(2) and 1.6 x 10(2) cfu/g, respectively. Isolates from both samples were identified to be Aspergillus flavus based on spore type and conidial structure of isolates. An increasing trend in the AFC of Cavendish bananas was observed during dried banana chips processing. Variability in the AFC between production batches was attributed to differences in aerobic and fungal populations and physicochemical characteristics of the fruits, peel damage of the raw materials, concentration of AFC in the air and food-contact surfaces of the production area, and temperature and relative humidity (RH) conditions of the environment during production and storage. Physicochemical characteristics of Cavendish bananas from the receipt of raw materials up to the first day of drying were within the reported range of values allowing growth and toxin production by aflatoxigenic fungi. Air-borne AFC varied depending on the section of the production area examined. The close proximity of the waste disposal area from the production operation to the preparation, drying and storage areas suggests that cross-contamination, probably air-borne or insect-borne was a likely occurrence. The hands of workers were also identified as AFC sources. Results of this study highlight the need for the development of strategies to control aflatoxigenic fungi and aflatoxin contamination in Philippine dried Cavendish bananas.

Menopause in Australia and Japan: effects of country of residence on menopausal status and menopausal symptoms.

OBJECTIVES: The aim of the study was to explore the effects of country of residence on menopausal status and menopausal symptoms in Australian and Japanese women. The study objectives included exploring the impact of country of residence (Australia and Japan) and menopausal status on menopausal symptoms, and identifying whether country of residence (Australia and Japan) moderates the relationship between menopausal status and menopausal symptoms. METHODS: Analyses are based on 1743 women aged between 40 and 60 years who participated in the multi-race, multi-site, cross-sectional study of mid-aged women called the Australian and Japanese Midlife Women's Health Study (AJMWHS) in 2001--2002. Study participants completed a mailed questionnaire that contained questions on a variety of health-related topics. RESULTS: In both cultures there was a similar increase in prevalence of depression (p < 0.001), somatic symptoms (p < 0.001) and vasomotor symptoms (p < 0.001) at perimenopause. Australian women experienced more night sweats than Japanese women but the prevalence of hot flashes was not statistically different. Postmenopausal Japanese women had more somatic, psychological and sexual symptoms. The main effect for menopausal status and the interaction effect of country of residence was significant in the somatic symptoms (p < 0.001), but not in any of the other areas. CONCLUSIONS: Vasomotor, psychological and somatic symptoms decrease after menopause in Australian women, with only sexual symptoms continuing. In Japanese women, somatic, psychological and sexual symptoms remain high after menopause. It is possible that westernization may be having a significant impact on the aging of women in Japan and it is, therefore important to capture through research just what this impact may be.

Occurrence of aflatoxin M1 in domestic milk in Japan during the winter season.

A total of 208 samples of commercial pasteurized milk gathered from retail outlets across Japan during the winter season were analysed for aflatoxin M1 (AFM1). Japan was divided into 11 regions from north to south, and nine to 45 milk samples from each region were randomly purchased between December 2001 and February 2002. Each milk sample was cleaned up by an immunoaffinity column, and AFM1 was quantified by liquid chromatography with fluorescence detection in four independent laboratories. The limit of detection of the method was 0.001 microg kg(-1). The identity of the putative AFM1 in milk sample was confirmed by the formation of AFM1 hemi-acetal with trifluoroacetic acid. Based on the results obtained with spiked samples (0.05 microg AFM1 kg(-1)), the mean recovery was 91.4%, the relative standard deviation for repeatability was 4.6%, and the relative standard deviation for reproducibility was 8.0% among four independent laboratories. AFM1 was detected in 207 (99.5%) of 208 milk samples at 0.001-0.029 microg kg(-1), with a mean of 0.009 microg kg(-1) and a 90th percentile of 0.014 microg kg(-1). No significant difference of the level of AFM1 contamination was observed among the regions.

In-house direct cELISA for determining aflatoxin B1 in Thai corn and peanuts.

To overcome the problem of aflatoxins (AF) in Thai foods, a sensitive in-house direct cELISA using monoclonal antibody (mAb) was established and compared with a commercial ELISA kit and thin-layer chromatography for the determination of AFB(1) levels in corn and peanuts. Among eight in-house mAbs (AF1-8), AF5 was used in the direct cELISA owing to its excellent specificity and sensitivity with the detection limit of 4 microg kg(-1). The recovery of AFB(1) spiked at 5, 10, 20, 40 and 80 microg kg(-1) ranged from 88.1 to 99.5%. Correlation coefficients of the ELISA with the commercial ELISA kit and thin-layer chromatography were 0.912 and 0.802 for corn, and 0.941 and 0.832 for peanuts, respectively (p<0.05). The cost per sample was estimated to be about 16 times lower than the commercial ELISA kit. Subsequently, the in-house direct cELISA was successfully applied to screen the contamination of AFB(1) in Thai corn and peanuts. Mean levels of AFB(1) (per cent positive) were 73 microg kg(-1) (85.7%) in corn and 102 microg kg(-1) (67.9%) in peanuts, for which 46.4% of both foods were above the Thailand regulation limit (20 microg kg(-1)).