Abstract. Previous our studies have shown that CD44, the principal receptor for hyaluronan, is present on cumulus cells during oocyte maturation. Although hyaluronan-CD44 interaction has been implicated in cumulus expansion and/or oocyte maturation, the full significance of CD44 remains unknown. The objective of the present study was to further investigate the role of CD44 in cumulus expansion and oocyte maturation in pigs. We demonstrate here in that CD44 has a key role in oocyte maturation but not in cumulus expansion. Previous studies have reported the physiological significance of cumulus expansion in oocyte maturation. However, our results suggest that cumulus expansion is a necessary condition for oocyte maturation, but that it is not sufficient on its own. Furthermore, western blot analysis demonstrated that the CD44 of the in vitro-matured cumulusoocyte complexes (COCs) had a larger molecular weight and more terminal sialic acid, which has been proven to inhibit the hyaluronan-binding ability of the receptor, than the CD44 of the in vivo-matured COCs, indicating that the hyaluronan-CD44 interactions during in vitro maturation might be insufficient compared with those in vivo. The insufficient interactions of hyaluronan-CD44 during in vitro maturation may cause the inferior capacity of fertilization and development of oocytes matured in vitro.

Macrophages are essential in cleaning up apoptotic debris during follicular atresia. However, the key factors of this process are still unclear. In the present study, we evaluated CD44 mRNA, CD44 protein, and CD44 antigen glycosylation on macrophages during follicular atresia in the pig. Atresia was classified into five stages: stage I, healthy follicles; stage II, early atretic follicles having apoptotic granulosa cells with an unclear basement membrane; stage III, progressing atretic follicles having apoptotic granulosa cells completely diffused from the basement membrane; stage IV, late atretic follicles with increasing lysosomal activity; and stage V, disintegrated atretic follicles having collapsed theca cells and strong lysosomal activity. Immunohistological analysis showed that macrophages expressing CD44 invaded the inside of stage III follicles, accompanied by a collapse of basement membrane. Semiquantitative RT-PCR showed that only mRNA of the CD44 standard isoform (CD44s) was present in inner cells of follicles, and not any CD44 variant isoform (CD44v) mRNAs. The amount of CD44s mRNA was increased at stage III. Western blot and lectin blot analyses showed that CD44 was markedly expressed at stage III and glycosylated with polylactosamine at the same time. After macrophages invaded atretic follicles at stages III-V, the CD44 expressed on macrophages was glycosylated with polylactosamine. The lysosomal activity began to increase at stage IV, and reached the highest level at stage V. Increased CD44s protein and posttranslational modification of CD44 with polylactosamine on macrophages from stage III could be involved in the cleaning up apoptotic granulosa cells.

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Background: Noncontact Electro Capacitive Cancer Therapy (ECCT) is a novel treatment modality in cancer. Chemokine (C-C motif) ligand 2 (CCL2) has a major role in the outgrowth of metastatic breast cancer. Interleukin 18 (IL18) plays a role in macrophage alteration, which leads to excessive angiogenesis. This study aims to elaborate on the association of CCL2, IL18, IL23α, and TNF-α (tumor necrosis factor-alpha) expression with the anti-proliferative effect of ECCT in rat breast tumor tissue.   Methods: Low intensity (18 Vpp) and intermediate frequency (150 kHz) alternating current-electric field (AC-EF) between two capacitive electrodes were exposed as external EF to a rat cage. Twenty-four rats were divided into four groups of six replicates. Breast tumor tissues were collected from 7, 12-dimethylbenz[a]anthracene (DMBA)-induced rats. Two groups were non DMBA-induced rats without ECCT exposure (NINT) and with (NIT). The other two groups were DMBA-induced rats without ECCT exposure (INT) and with (IT). Mammary glands and breast tumor tissues were collected from each group and preserved. Hematoxylin-eosin and immunohistochemistry staining were performed on paraffin sections of tissues using anti-PCNA, anti-ErbB2, anti-Caspase3, and anti-CD68. CCL2, IL18, IL23α, and TNF-α mRNA relative expressions were analyzed using qRT-PCR. Results: ECCT exposure may cause the reduction of PCNA protein expression as well as ErbB2 on breast tumor tissues, but it causes the increase of Caspase3 and macrophage CD68 protein. In rat breast tumor tissues of IT groups, the mRNA expression of CCL2 and IL18 are significantly down-regulated, in contrast with the up-regulated expression of these cytokines in tumor tissues of the INT group. IL23α and TNF- α expression remained similar in both groups. Conclusion: CCL2 and IL18 expressions have an association with the inhibition of breast tumor cell proliferation affected by ECCT exposure

The chemical contents and health benefits of black rice bran of some rice cul vars have been inves gated. However, there has been li le research on the 'Cempo Ireng' cul var from Sleman, Yogyakarta. The aim of this present study was to determine the anthocyanin, an oxidant ac vity, and macro-and micronutrients contents of black rice bran from this local cul var. The anthocyanin in the black rice bran was extracted using the macera on method with methanol as a solvent. The extract obtained was separated through a prepara ve thin layer chromatography (TLC) of silica GF254 and a mobile phase composed of n-butanol, ace c acid, and water. Two frac ons were collected and analyzed for the anthocyanin content. The prepara ve TLC spots were separated for further detec on and measurement of cyanidin 3-O-glucoside using HPLC followed by LC-MS. The an oxidant ac vity of the frac ons were measured using the DPPH free radical scavenging method. The results showed that the anthocyanin in frac on 1 was iden fied as cyanidin 3-O-glucoside (66.1 ± 10.6 µg/g). The IC 50 of frac ons 1 and 2 were 200.96 and 218.36 µg/mL, respec vely. Analysis of the macro-and micronutrients revealed that the black rice bran of 'Cempo Ireng' had nutrient contents comparable with other rice cul vars. Therefore, this local black rice bran can be used as a source of an oxidants and macro-and micronutrients.

Previous study showed that kaffir lime leaf contains alkaloid, flavonoid, terpenoid, tannin and saponin. The objective of this study was to examine the cytotoxic effect of kaffir lime leaf extract on cervical cancer and neuroblastoma cell lines. The method used for this research to determine cell viability was an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results showed that an ethyl acetate extract had an IC50 for HeLa cells, UKF-NB3, IMR-5 and SK-N-AS parental cells of 40.7 μg · mL -1 , 28.4 μg · mL -1 , 14.1 μg · mL -1 , and 25.2 μg · mL -1 respectively. Furthermore, the IC50 of chloroform extracts for HeLa cells, UKF-NB3, IMR-5 and SK-N-AS parental were 17.6 μg · mL -1 , 18.9 μg · mL -1 , 6.4 μg · mL -1 , and 9.4 μg · mL -1 respectively. These data showed that kaffir lime extract reduces the viability of cervical and neuroblastoma cell lines and may have potential as anti-cancer compounds. © 2015 W.A.S. Tunjung, J. Cinatl, M. Michaelis, C.M. Smales. Published by Elsevier B.V. Peer-review under responsibility of the Scientific Committee of HK-ICONS 2014.

Abstract. Hyperuricemia can be induced by the high level of uric acid in the blood. Ginger flower (E. elatior Jack.) contains flavonoid, phenol and glycoside. Polyphenol and flavonoid have been reported having Xantine Oxidase Inhibitor (XOI) activity that can help for decreasing uric acid levels. Hence, the ginger flower can be used as a candidate of traditional hyperuricemia medicine. The objective of this study was to analyze the ability of ginger flower extract to decrease uric acid level in the hyperuricemic rat. Methods of this study were sample collection and preparation of ginger flower extract by infundation method, induction of hyperuricemic rat using per oral administration of beef broth at dose of 700 mg/kg body weight, treatment of hyperuricemic rat using ginger extract at dose of 200 mg/kg body weight and allopurinol as uric acid drug at dose of 180 mg/kg body weight and measurement of uric acid concentration. Results showed that ginger flower extract decreased uric acid concentration until 31.78% on discontinuous hyperuricemic induction sub-group and 17.90% on continuous hyperuricemic induction sub-group. Allopurinol decreased uric acid concentration until 45.65% on discontinuous hyperuricemic induction sub-group and 23.53% on continuous hyperuricemic induction sub-group. Hence, the ginger flower had the ability to decrease uric acid concentration in hyperuricemic rat into the normal level.

Previous studies have reported that a number of organic compounds are present in kaffir lime (Citrus hystrix DC.) leaf extracts. Further research is needed to purify these compounds and determine which are biologically active. The objective of this study is to identify the volatile organic compounds of kaffir lime leaf crude extracts and fractions and to study their bioactivity. Fractionation was performed by the double maceration method, using hexane as the second solvent. TLC was performed to analyze the qualitative separation, whereas the individual constituents were detected using GC-MS. Our results showed that chloroform and ethyl acetate crude extracts contained various volatile organic compounds such as fatty acids, fatty alcohols, prenol lipids, sterol lipids, terpenoids and long chain alkanes. Fractionation separated these compounds into non-hexane fractions, which contained less volatile compounds, and hexane fractions. The volatile compounds of non-hexane fractions were identified to be long chain alkanes, meanwhile the hexane fractions contained terpenoids, fatty acids, fatty alcohols, prenol lipids and sterol lipids. Palmitic acid and terpenoids, such as citronellyl propionate, nerolidol, citronella and caryophyllene oxide were found to be the most dominant bioactive compounds in chloroform and ethyl acetate crude extract and their hexane fractions, which were reported to possess cytotoxicity against cancer cells. Meanwhile in non-hexane fractions, long chain alkanes such as triacontane and hentriacontane were found to be the most dominant bioactive compound which also possessed cytotoxic effect. In conclusion, fractionation using the double maceration method yielded different volatile organic compounds composition with different biological activities. The crude extracts and fractions of kaffir lime leaves were potential to be developed as a traditional medicine for cancer treatment.