Role of Runx2 phosphorylation in prostate cancer and association with metastatic disease.

The osteogenic transcription factor, Runx2, is abnormally expressed in prostate cancer (PCa) and associated with metastatic disease. During bone development, Runx2 is activated by signals known to be hyperactive in PCa including the RAS/MAP kinase pathway, which phosphorylates Runx2 on multiple serine residues including S301 and S319 (equivalent to S294 and S312 in human Runx2). This study examines the role of these phosphorylation sites in PCa. Runx2 was preferentially expressed in more invasive PCa cell lines (PC3>C4-2B>LNCaP). Furthermore, analysis using a P-S319-Runx2-specific antibody revealed that the ratio of P-S319-Runx2/total Runx2 as well as P-ERK/total ERK was highest in PC3 followed by C4-2B and LNCaP cells. These results were confirmed by immunofluorescence confocal microscopy, which showed a higher percentage of PC3 cells staining positive for P-S319-Runx2 relative to C4-2B and LNCaP cells. Phosphorylated Runx2 had an exclusively nuclear localization. When expressed in prostate cell lines, wild-type Runx2 increased metastasis-associated gene expression, in vitro migratory and invasive activity as well as in vivo growth of tumor cell xenografts. In contrast, S301A/S319A phosphorylation site mutations greatly attenuated these Runx2 responses. Analysis of tissue microarrays from 129 patients revealed strong nuclear staining with the P-S319-Runx2 antibody in primary PCas and metastases. P-S319-Runx2 staining was positively correlated with Gleason score and occurrence of lymph node metastases while little or no Runx2 phosphorylation was seen in normal prostate, benign prostate hyperplasia or prostatitis indicating that Runx2 S319 phosphorylation is closely associated with PCa induction and progression towards an aggressive phenotype. These studies establish the importance of Runx2 phosphorylation in prostate tumor growth and highlight its value as a potential diagnostic marker and therapeutic target.

Role of melanin in mechanical properties of Glycera jaws.

The remarkable mechanical prowess of the jaws of the bloodworm Glycera dibranchiata appears to be a consequence of a robust cross-linked network of organic molecules, notably melanin and proteins, as well as small amounts of unmineralized Cu and a Cu-based mineral. The present study focuses on the role of melanin. Mechanical properties of untreated jaws and the constituent melanin are probed through nanoindentation, both in air and underwater. Complementary information is obtained from density and porosity measurements and attempts at Cu removal from the jaws using EDTA, an effective metal chelator in most biological systems. In near-tip regions of the jaws, mechanical properties attain the highest values and diminish only slightly when wet (by 15-25%), in contrast to the behavior of other organic biomaterials. The melanin constituent contributes significantly to the mechanical integrity of the jaw; its hardness and elastic modulus are about half those of untreated jaws. Although melanin may be the dominant shape-determining component of the structure, it remains to be shown whether jaw assembly is mediated by protein deposition on a melanin scaffold or, conversely, by melanin deposition on a protein scaffold. The inability of EDTA to chelate Cu from the jaws and the relatively high density of the jaws and the melanin support the notion of a highly cross-linked molecular structure. Finally, based on the metric H(3)/E(2) (H being hardness and E the Young's modulus), the results suggest that the abrasion resistance of the jaws is superior to all engineering polymers and competitive with the hardest metallic alloys.

A comparative study on the oestrous response to PGF2 alpha analogue treatment, and conception rates according to time of artificial insemination, in Zebu (Bos indicus) and Baoulé (Bos taurus) cattle.

The oestrous response, interval and conception rates were studied after synchronization with a prostaglandin analogue (cloprostenol) and artificial insemination (AI) performed at different times in 50 Zebu (Bos indicus) and 83 Baoulé (Bos taurus) cattle indigenous to Burkina Faso. The overall proportion of cows responding to synchronization was 70% (93/133). Although the response was higher for the Baoulé cattle, at 73.5% (61/83), than for the Zebu, at 64% (32/50), the difference was not statistically significant (p > 0.05). The mean oestrous interval from treatment to the onset of oestrus (TOI) was shorter in the Zebu (54.1 h, SD 6.7) than in Baoulé (65.2 h, SD 12.9) cattle (p < 0.001). Of the Zebu (n = 32) that responded, 65.7% presented oestrus over a period of 12 h ranging from 48 h to 60 h after treatment. For the Baoulé cows, the highest proportion of animals in oestrus over a period of 12 h was 41% between 60 h and 72 h after treatment. The frequency distribution of onset of the oestrus indicated that up to 64.5% of the Zebu and 79.5% of the Baoulé cattle showed onset of oestrus during the daytime. For Zebu and Baoulé cows inseminated 13 h or 18 h after the onset of oestrus, conception rates were 56% and 57% (p > 0.05) and 33% and 64% (p < 0.05), respectively. Based on these findings, it appears that the oestrous response to synchronization was adequate for both Zebu and Baoulé cattle and that the time to onset of oestrus varied according to genotype. It was also concluded that conception rates were satisfactory for both genotypes but that, for Baoulé cattle, AI performed 18 h after oestrus significantly increased conception rates compared to AI at 13 h after oestrus.

The reproductive performance of East African (Bos Indicus) Zebu cattle in Ethiopia. 1. Estrous cycle length, duration, behavior and ovulation time.

The average (+/- standard deviation) estrous cycle length of 28 East African Zebu cows over a 217-d period, was 22.6 +/- 6.5 d with no significant (P>0.05) difference between seasons. Estrus had a mean duration of 7.66 +/- 4.68 h (ranging from 1 to 24 h) followed by ovulation 25.82 +/- 5.25 h after the onset of estrus. A larger number of estruses started during the day (64 vs 36% P<0.001) and they were longer during the dry season (P<0.05). Proestrus and metaestrus had average duration of 3.46 +/- 3.57 and 3.65 +/- 2.87 h, respectively. Of the estruses recorded, 31% had no proestrus and 34% had no metaestrus. More mounting occurred during the day than night (59 vs 41%; P<0.001), and mounting activity had two peaks: 0600 to 0900 h and 1600 to 1900 h. The average number of mounts observed per estrus was 9.2 (ranging from 1 to 58), and the mounts were concentrated at the beginning and end of estrus, irrespective of their duration. Vaginal mucus discharge was detected in 64% of the cows in estrus.