New piericidin rhamnosides as potentiators of amphotericin B activity against Candida albicans produced by actinomycete strain TMPU-A0287.

Four new piericidin rhamnosides (2, 4-6) together with three known piericidins (1, 3, 7) were isolated from the culture broth of the unidentified actinomycete strain TMPU-A0287 as potentiators of antifungal amphotericin B (AmB) activity. The structures of piericidins were elucidated by spectroscopic analyses, including NMR and MS. Compounds 2 and 4-6 possessed a ketone at C-10 and one or two methoxy groups on the rhamnose in their structures. Compounds 1-7 did not exhibit antifungal activity against Candida albicans and all potentiated AmB activity. The MIC values of AmB against C. albicans combined with 1-7 (4.0 µg ml-1) decreased from 0.50 to 0.063 or 0.031 µg ml-1, yielding an 8- or 16-fold increase in AmB activity.

Using the Barthel Index to Assess Activities of Daily Living after Musculoskeletal Tumour Surgery: A Single-centre Observational Study.

OBJECTIVE: The objective of the current study was to find the factors affecting the activities of daily living, as evaluated by the Barthel Index, at the end of rehabilitation after musculoskeletal tumour surgery. Further, we evaluated whether the Barthel Index correlates with functional scores that are specific to musculoskeletal tumours at final follow-up. METHODS: The activities of daily living of 190 patients who underwent postoperative rehabilitation after surgery to treat musculoskeletal tumours were evaluated at the end of the program using the Barthel Index. Functional evaluation at the time of final follow-up observation was evaluated using the Musculoskeletal Tumour Society Score and the Toronto Extremity Salvage Score. RESULTS: The post-rehabilitation Barthel Index was significantly lower in elderly patients aged more than 60 years and in those with malignant tumours and tumours larger than 10 cm. Malignancy and large tumour size were risk factors for a low Barthel Index. There was significant correlation between the Musculoskeletal Tumour Society Score/Toronto Extremity Salvage Score at final functional evaluation and the Barthel Index at the end of rehabilitation. CONCLUSION: The Barthel Index is a simple method to assess the activities of daily living and can potentially predict disease-specific health-related quality of life at final functional evaluation after musculoskeletal tumour surgery.

Guava leaf extract suppresses osteoarthritis progression in a rat anterior cruciate ligament transection model.

Guava leaf extract and ellagic acid, one of its polyphenolic components, inhibit the activity of a disintegrin and metalloproteinase with thrombospondin type 5 (ADAMTS-5), which is associated with aggrecan degeneration during the early stage of osteoarthritis (OA). To investigate the efficacy of guava leaf extract for preventing OA, we examined the effect of its dietary intake on cartilage destruction in anterior cruciate ligament-transected (ACLT) rats. Rats were randomly assigned to four groups: ACLT control rats fed with control diet, ACLT rats fed with diet containing 0.2% guava leaf extract, ACLT rats fed with diet containing 0.5% guava leaf extract, and sham-operated rats fed with control diet. Mankin's scores, an index of cartilage damage, were higher in rats that underwent ACLT. Guava leaf extract treatment dose-dependently led to lower Mankin's scores and higher concentrations of ellagic acid in the serum and synovial membrane. Ellagic acid levels in the synovial membrane negatively correlated with cartilage destruction scores. These results suggest that dietary guava leaf extract suppresses OA progression in ACLT rats through ellagic acid-mediated inhibition of early joint destruction.

Effects of Psidium guajava Linn. leaf extract in Japanese subjects with knee pain: a randomized, double-blind, placebo-controlled, parallel pilot study.

The extract of Psidium guajava Linn. (guava) leaf was recently revealed to suppress the advance of osteoarthritis (OA) in rat anterior cruciate ligament-transection models. To investigate the efficacy of guava leaf extract in improving knee pain, which is a common symptom of OA, we conducted a double-blind parallel pilot clinical study in Japanese subjects with knee pain. The subjects, who had no medical history of knee treatment, were randomly assigned to two groups with similar total Japanese Knee Osteoarthritis Measure (JKOM) scores. During the 12-week intake period, the subjects in each group ingested 1 g of guava leaf extract (the guava group) or placebo (the placebo group) daily. At week 12, pain and stiffness in knees (one subcategory of JKOM score) in the guava group was significantly lower than that in the placebo group. Using the visual analogue scale (VAS) for knee pain, a significant association between treatment effect and test period was shown, and the guava group had a lower VAS score at week 12 than the placebo group. In conclusion, continuous intake of guava leaf extract might relieve knee pain, suggesting a potential preventive effect against OA symptoms.

Purification, Characterization, and Gene Cloning of a Cold-Adapted Endo-1,4-ß-glucanase from Bellamya chinensis laeta.

An endo-1,4-ß-glucanase from Bellamya chinensis laeta was purified to electrophoretically homogeneous state. The molecular weight of the purified enzyme was estimated 70,000 by SDS-PAGE. The enzyme was most active at pH 5.5 and 50 °C, and stable at around pH 10 and 50 °C. The enzyme exhibited the significant activity at 20 °C (30 % of the activity at optimal 50 °C). The enzyme was hydrolyzed cellohexaose into cellobiose, cellotriose, and cellotetraose as main products. Three cDNAs (BC-EG70a, BC-EG70b, and BC-EG70c) encoding the endo-1,4-ß-glucanase were cloned by PCR-based method. Three endo-1,4-ß-glucanases consisted of 1758 bp encoding 586 amino acids. The three genes were almost the same nucleotide sequences. The deduced proteins were consisted of a signal sequence, cellulose binding domain, linker, and catalytic domain. The amino acid sequence of BC-EG70a shares sequence identity degree with the endo-1,4-ß-glucanases of Haliotis discus hannai (61 %), Ampullaria crossean (52 %), and Mizuhopecten yessoensis (51 %) which all belong to glycoside hydrolase family 9.

Significance of beta-catenin and pRB pathway components in malignant ovarian germ cell tumours: INK4A promoter CpG island methylation is associated with cell proliferation.

To clarify the mechanisms underlying cell cycle promotion in malignant germ cell tumours of the ovary (MGCTOs), beta-catenin and components of the pRB pathway, cyclin D1 and p16, were analysed in relation to cell proliferation. Immunohistochemically, p16 protein was not expressed in a number of MGCTOs (9 of 42 tumours: 21.4%) and was associated with p16 gene (INK4A) promoter 5'-CpG islands methylation. Amplification of the cyclin D1 gene (CCND1) was detected in a small number of MGCTOs (5 of 42 tumours: 13.5%). Reduced expression of p16 due to promoter methylation correlated significantly with increased cell proliferation as evidenced by Ki-67 labelling index (p < 0.001) and mitotic index (p < 0.01). In some tumour types, nuclear localization of beta-catenin has been reported to be associated with beta-catenin gene (CTNNB1) mutation, cyclin D1 overexpression, and increased cell proliferation. Nuclear localization of beta-catenin, which was observed in MGCTOs other than dysgerminoma, was not associated with cyclin D1 expression and increased cell proliferation, but appeared to be related to tumour differentiation. Furthermore, CTNNB1 mutations were not detected in any of the MGCTOs examined. Our results suggest that reduced expression of p16 due to INK4A promoter methylation is one of the principal factors that promote cell proliferation in MGCTOs. Thus, p16 may be a novel target for gene therapies to treat MGCTOs.

11q23-24 loss is associated with chromosomal instability in endometrial cancer.

A loss of the DNA copy number at chromosomal region 11q23-24 as detected by comparative genomic hybridization (CGH) is a marker of poor prognosis in patients with endometrial cancer. Malignant tumors display genetic instability, which is classified into two types: microsatellite instability (MIN) and chromosomal instability (CIN). In the present study, we examined whether there is a relation between loss of 11q23-24 and genetic instability in endometrial adenocarcinoma. Loss of 11q23-24 was detected in 4 of 70 endometrial cancers by fluorescence in situ hybridization (FISH), and DNA aneuploidy was detected by laser scanning cytometry (LSC) in 14 tumors. All tumors with 11q23-24 loss were aneuploid, and three of them were considered to have CIN. These findings suggest that 11q23-24 contains gene(s) necessary for normal chromosome replication and cell division.

Genetic alterations related to lymph node metastasis and peritoneal dissemination in epithelial ovarian cancers.

Genetic alterations are assumed to be necessary for the development and progression of ovarian cancer. However, the genetic alterations that occur during lymph node metastasis and peritoneal dissemination are poorly understood. In the present study, we used comparative genomic hybridization to detect genetic alterations in 30 tumors from patients with primary ovarian cancers and analyzed the associations of these genetic alterations with clinical stage and surgical pathological factors, such as histological grade, lymph node metastasis, and peritoneal dissemination. The total number of genetic alterations per tumor ranged from 0 to 39, with an average of 17.7 alterations per tumor. Among the genetic alterations in ovarian cancers, gains on chromosomes 8q and 3q and losses on chromosomes 17p, 18q, and 4q were observed frequently. Although the difference in total numbers of genetic alterations between early-stage tumors and advanced-stage tumors was not significant, the difference was significant when high-grade cancers were compared with low-grade cancers. Eight regions on seven chromosomes showed genetic alterations related to lymph node metastasis or peritoneal dissemination. Gain at 11q13-q14 and loss at 17q11.2-q21 were related not only to lymph node metastasis and peritoneal dissemination but also to clinical stage and histological grade.

Alpha-monoisostearyl glyceryl ether enhances percutaneous penetration of indometacin in-vivo.

Molecules that reversibly remove the barrier resistance of skin enhance penetration. alpha-Monoisostearyl glyceryl ether (GE-IS) is a novel compound that can be used as a non-ionic surfactant and increases percutaneous penetration of indometacin in rat abdominal skin in-vitro. The present study investigated GE-IS-induced enhancement of indometacin penetration in-vivo. When 1% GE-IS in propylene glycol was applied to rat abdominal skin, serum and muscle concentrations of indometacin increased markedly. Anti-inflammatory activities of test solutions containing both indometacin and GE-IS were investigated in experimental models of acute and chronic inflammation. Application of indometacin with GE-IS to the skin produced greater inhibitory effects on carrageenan-induced rat paw oedema, UV-induced erythema in guinea-pigs, and adjuvant arthritis in rats, compared with application of indometacin alone. The results suggest that GE-IS enhances penetration in-vivo and improves the anti-inflammatory effects of indometacin in animal models. Thus, GE-IS might contribute to the development of cosmetic or medical formulations to improve transfer of bioactive substances to hypodermal sites.

Syndecan-1 expression in cancer of the uterine cervix: association with lymph node metastasis.

The development of carcinoma is associated with alterations in the expression of many cell adhesion molecules. Syndecan-1 is a cell surface proteoglycan that binds cells to the extracellular matrix and changes its expression following malignant transformation in some tumors. Our purpose was to examine the pattern of syndecan-1 expression in cancer of the uterine cervix and assess the clinicopathological significance of syndecan-1 expression. A total of 106 tissue specimens (6 normal, 19 cervical intraepithelial neoplasia (CIN) and 81 invasive cancer) were analyzed immunohistochemically. In addition, the corresponding expression of mRNA in tumor tissues was evaluated by reverse transcription-polymerase chain reaction (RT/PCR) in comparison with normal counterparts. Syndecan-1 was positive in normal squamous cells except the basal cell layer. The intensity of syndecan-1 staining was the strongest in normal epithelium, followed by CIN, and invasive squamous cell carcinoma. Syndecan-1 expression in cancer tissue tended to be higher in keratinizing type than non-keratinizing type and not found in adenocarcinoma. Syndecan-1 expression was markedly decreased at the mRNA level in invasive squamous cell carcinoma as compared with that of normal uterine cervix. Interestingy, there was an inverse correlation between the expression of syndecan-1 in the primary site and lymph node metastasis, although there was no significant correlation between syndecan-1 expression and the prognosis. The results of the present study suggest that syndecan-1 expression is associated with squamous tissues and plays a key role in the progression of the cancer of the uterine cervix especially in the metastatic process.