Avidin related protein 2 shows unique structural and functional features among the avidin protein family

Background  

The chicken avidin gene family consists of avidin and several avidin related genes (AVRs). Of these gene products, avidin is the best characterized and is known for its extremely high affinity for D-biotin, a property that is utilized in numerous modern life science applications. Recently, the AVR genes have been expressed as recombinant proteins, which have shown different biotin-binding properties as compared to avidin.     

Androgen receptor mRNA under-expression in poorly differentiated human hepatocellular carcinoma

Many studies suggest that hepatocellular carcinoma (HCC) is an androgen-dependent tumor with an incidence five times higher in males, but few data are available on the androgen receptor (AR) mRNA levels in different physiological classes of human liver specimens. In this study 108 human hepatic samples have been analyzed for AR mRNA expression by a comparative RT-PCR assay. These consisted of 35 non-tumoral hepatic samples (3 normal parenchymas, 4 steatosis, 10 hepatitis, 18 cirrhosis), 38 tumoral specimens derived from uninodular and multinodular HCCs and 35 peritumoral hepatic tissues. Normalized AR mRNA levels in tumoral and peritumoral liver tissues spanned from 0 to 146% and from 7 to 125% respectively. Only in a relatively small percentage of HCCs, the levels of expression of AR mRNA were higher than in the corresponding peritumoral tissues (16% of total HCCs). Although extremely variable, the AR mRNA levels were related to histological tumoral differentiation and proved to be lower in the highly dedifferentiated HCCs as compared to the well differentiated ones. Therefore, the evaluation of AR expression in HCC patients might be relevant for the planning of clinical studies on anti-androgen therapies, which might be useful only in the cases in which a high level of AR mRNA is detected, considering the high heterogeneity of AR mRNA levels which characterizes HCC samples. It is likely that the HCCs, expressing low or undetectable levels of AR mRNA, would not benefit by the anti-androgen therapy.     

Induction of fibronectin mRNA by urokinase- and tissue-type plasminogen activator in human skin fibroblasts: differential role of u-PA and t-PA at the fibronectin protein level

Plasminogen activators of the urokinase- and tissue-type and fetal calf serum (u-PA, t-PA, FCS) exert their mitogenic effect on quiescent human dermal fibroblasts and modulate the mRNA expression of cell-cycle related genes. The present study deals with the effects of PAs on the expression of fibronectin (FN), a heterodimeric extracellular matrix (ECM) protein that can be modulated in different ways by various mitogens. The kinetics of FN gene response was examined in quiescent fibroblasts upon PA stimulation (30 min -24 h). The results obtained evidenced that: (i) all mitogens tested (u-PA, t-PA and FCS) led to an increase of FN mRNA expression in early G1, as shown by the analysis of two sequences, III-9, common to all FN mRNAs, and EDA+, present only in the EDA+FN isoform; (ii) the kinetic profiles of FN mRNA stimulation were comparable for the three mitogens, although the effects on the FN-ECM assembly were distinct; (iii) t-PA and FCS led to FN assembly in the ECM, which was absent or decreased in u-PA-treated cultures. Immunobiochemical analysis of total FN and EDA+ FN showed that FN induced by t-PA was mainly dimeric (450-500 kDa), whereas FN induced by u-PA was mainly monomeric (230-250 kDa). These differences are probably due to the differential enzymatic action of t-PA and u-PA on FN, which might be related to a differential role of the two PAs in several physiopathological conditions.     

Optimizing Controlled Environment Assessment of Levels of Resistance to Yam Anthracnose Disease Using Tissue Culture-derived Whole Plants

Studies were conducted to determine the timing and frequency of disease assessment required to effectively identify levels of resistance to yam anthracnose using tissue culture‐derived whole plant inoculation assay. The effects of inoculation methods (paint brush and spray), and disease scoring methods [individual leaf area (ILA) and whole plant area (WPA)] were also assessed. Spray inoculation resulted in rapid infection and higher variations among yam genotypes, leading to earlier discrimination of genotypes than with the paintbrush method. Both the ILA and WPA scoring methods showed variation among yam genotypes, and association between the two methods gave a high positive correlation (r > 0.90). However, the WPA was faster and had the advantage of detecting differences in reactions of yam genotypes to less aggressive pathogen isolates to which the ILA method showed no variation. A single disease evaluation at 7 days after inoculation was as good as the area under the disease progress curve (AUDPC) and the disease progress rate (R_d) derived from multiple evaluations. However, a significant time–genotype interaction, suggests a need for more than a single assessment for effective comparison of genotypes. AUDPC derived from two assessments (5 and 7 DAI) was better than AUDPC from three assessments (5, 7 and 9 DAI) in separating genotypes reactions to a less aggressive pathogen isolate. This study showed that the use of spray inoculation method, the WPA scoring method, and AUDPC derived from two assessments (5 and 7 DAI) provided best conditions for evaluating yam genotypes for levels of anthracnose resistance with the tissue culture‐derived whole plant assay.     

Response of hepatic metallothionein to iron administration

Female broiler chicks receiving an ip injection of Fe (10 mg/kg bw) were found to have a greatly increased level of hepatic metallothionein. The increase in metallothionein was not correlated with changes in serum corticosterone. Attempts to vary serum corticosterone levels by feeding metyrapone, an 11-β steroid hydroxylase inhibitor, were unsuccessful.     

Straightforward hit identification approach in fragment-based discovery of bromodomain-containing protein 4 (BRD4) inhibitors

A combination approach of a fragment screening and “SAR by catalog” was used for the discovery of bromodomain-containing protein 4 (BRD4) inhibitors. Initial screening of 3695-fragment library against bromodomain 1 of BRD4 using thermal shift assay (TSA), followed by initial hit validation, resulted in 73 fragment hits, which were used to construct a follow-up library selected from available screening collection. Additionally, analogs of inactive fragments, as well as a set of randomly selected compounds were also prepared (3?×?3200 compounds in total). Screening of the resulting sets using TSA, followed by re-testing at several concentrations, counter-screen, and TR-FRET assay resulted in 18 confirmed hits. Compounds derived from the initial fragment set showed better hit rate as compared to the other two sets. Finally, building dose-response curves revealed three compounds with IC₅₀?=?1.9–7.4?μM. For these compounds, binding sites and conformations in the BRD4 (4UYD) have been determined by docking.