Calmodulin is essential for estrogen receptor interaction with its motif and activation of responsive promoter

Calmodulin (CaM) has been reported to have affinity for the estrogen receptor (ER). Observations reported here reveal a direct physical interaction between purified CaM and ER. This direct ER-CaM interaction may be an initial event preceding the assembly of ER plus auxiliary proteins into the active ER complex with its DNA motif, the estrogen response element. We demonstrate that CaM is an integral component of this complex by using a system reconstituted from purified ER and nuclear extract from ER-negative breast cancer cells and also with ER-depleted nuclear extract of an ER-positive breast cancer cell line. Although CaM is essential for formation of this complex, it is not sufficient, suggesting roles also of auxiliary proteins. CaM also is functionally required for activation of an ER-responsive promoter, in the 17beta-estradiol-ER pathway of hormone action and regulation of 17beta-estradiol-responsive gene expression that is associated with proliferation of mammary epithelial cells.     

A nonblocking transaction data flow graph based protocol forreplicated databases

Replicated data management systems adopt the 1-copy serializability criteria for processing transactions. In order to achieve this goal, many approaches rely on obtaining votes from other sites for processing update requests. In the proposed approach, a technique for generation of precedence graphs for each transaction execution is analyzed. The transaction data flow graph approach is a fully distributed approach. The proposed technique, is free from deadlocks, and avoids resubmission of transactions     

Identification of an interaction between the m-band protein skelemin and beta-integrin subunits. Colocalization of a skelemin-like protein with beta1- and beta3-integrins in non-muscle cells

Signaling across integrins is regulated by interaction of these receptors with cytoskeletal proteins and signaling molecules. To identify molecules interacting with the cytoplasmic domain of the beta3-integrin subunit (glycoprotein IIIa), a placental cDNA library was screened in the yeast two-hybrid system. Two identical clones coding for a 96-amino acid sequence were identified. This sequence was 100% identical to a sequence in skelemin, a protein identified previously in skeletal muscle. Skelemin is a member of a superfamily of cytoskeletal proteins that contain fibronectin-type III-like motifs and immunoglobulin C2-like motifs and that regulate the organization of myosin filaments in muscle. The amino acid residues in the isolated clones encompassed C2 motifs 4 and 5 of skelemin. A recombinant skelemin protein consisting of C2 motifs 3-7 interacted with beta1- and beta3-integrin cytoplasmic domains expressed as glutathione S-transferase (GST) fusion proteins, but not with GST-beta2-integrin cytoplasmic tail or GST alone. The skelemin-binding region was in the membrane proximal cytoplasmic domains of the integrins. Full-length skelemin interacted with integrin in intact cells as demonstrated by the colocalization of hemagglutinin-tagged skelemin in Chinese hamster ovary (CHO) cells containing alphaIIbbeta3-integrin and by the finding that microinjection of C2 motif 4 of skelemin into C2C12 mouse myoblast cells caused spread cells to round up. A skelemin-like protein was detected in CHO cells, endothelial cells, and platelets, and this protein colocalized with beta1- and beta3-integrins in CHO cells. This study suggests the presence of a skelemin-like protein in non-muscle cells and provides evidence that it may be involved in linking integrins to the cytoskeleton.     

Cloning of the cDNA for glutamyl-tRNA synthetase from Arabidopsis thaliana

High-sensitivity titration calorimetry is used to measure changes in enthalpy, heat capacity and protonation for the binding of captopril to the angiotensin I-converting enzyme (ACE; EC 3.4.15.1). The affinity of ACE to captopril is high and changes slightly with the pH, because the number of protons linked to binding is low. The determination of the enthalpy change at different pH values suggests that the protonated group in the captopril-ACE complex exhibits a heat protonation of approximately -30 kJ/mol. This value agrees with the protonation of an imidazole group. The residues which may become protonated in the complex could be two histidines existing in two active sites, which are joined to the amino acids coordinated to Zn2+. Calorimetric measurements indicate that captopril binds to two sites in the monomer of ACE, this binding being enthalpically unfavorable and being dominated by a large positive entropy change. Thus, binding is favored by both electrostatic and hydrophobic interactions. The temperature dependence of the free energy of binding deltaG degrees is weak because of the enthalpy-entropy compensation caused by a large heat capacity change, deltaCp =-4.3+/-0.1 kJ/K/mol of monomeric ACE. The strong favorable binding entropy and the negative deltaCp indicate both a large contribution to binding due to hydrophobic effects, which seem to originate from dehydration of the ligand-protein interface, and slight conformational changes in the vicinity of the active sites.     

Geographic variations in the composition of ivory of the African elephant (Loxodonta africana)

OBJECTIVES: The purpose of the present study was to compare the inclination of the occlusal plane with occlusal guidance as a contributing factor to masticatory movement. METHODS: Masticatory movements of 41 young adults were measured using a 3-D mandibular movement analysing system. The inclination of the occlusal plane was measured in the sagittal plane using a 3-D digitizer. The contribution of the occlusal guidance and the inclination of the occlusal plane to the direction of the masticatory path of closure was evaluated at various closing levels. RESULTS: The masticatory path of closure outside the intercuspal range was influenced mainly by the inclination of the occlusal plane, and the masticatory path of closure near the intercuspal range was only influenced by occlusal guidance. The so-called gliding type masticatory pattern was observed predominantly in subjects with a posteriorly inclined occlusal plane. In contrast, a chopping type masticatory pattern was observed predominantly in subjects with an anteriorly inclined occlusal plane. CONCLUSIONS: The contribution of the inclination of the occlusal plane to masticatory movement was greater than that of occlusal guidance throughout the closing phase except near the intercuspal range.     

Descriptive epidemiology of childhood cancers in Bangalore, India

While fairly complete and reliable incident data on childhood cancers are available from the registries in India, mortality and survival information is not. Information concerning the latter was obtained by the Bangalore cancer registry through active follow-up involving visits to homes of patients. Between 1982 and 1989, 617 cases of cancers in childhood were registered, giving an age-standardized incidence rate of 84.8 and 48.4 per million in male and female children, respectively. Active follow-up provided mortality/survival information in 532 or 86.2 percent of these cases. Overall, observed five-year survival was 36.8 percent (both genders combined) with a relative survival of 37.5 percent when childhood mortality in the general population was taken into account. The five-year relative survival was best for thyroid carcinoma (100 percent) followed by Hodgkin's disease (73 percent) and retinoblastoma (72.9 percent). Survival was comparatively low, being 9.9 percent in acute nonlymphatic leukemia and less than 20 percent in rhabdomyosarcoma and the category grouped as 'other malignant neoplasms.' Survival in Hodgkin's disease was influenced by clinical stage at presentation, but was not statistically significant possibly due to small numbers.     

Using time-driven activity-based costing to manage digital forensic readiness in large organisations

A digital forensic readiness (DFR) programme consists of a number of activities that should be chosen and managed with respect to cost constraints and risk. Traditional cost systems, however, can not provide the cost of individual activities. This makes it difficult or impossible for organisations to consider cost when making decisions about specific activities. In this paper we show that the relatively new cost system, time-driven activity-based costing (TDABC), can be used to determine the cost of implementing and managing activities required for DFR. We show through analysis and simulation that the cost information from a TDABC model can be used for such decisions. We also discuss some of the factors that ought to be considered when implementing or managing the use of TDABC in a large organisation.     

Microstructure refinement with forced convection in aluminium and superalloys

The cooling and average local solidification times were determined for slow solidifiation of Al-4.4 wt% Cu alloy under natural convection and under electromagnetically forced axisymmetric rotation during liquid cooling and solidification in graphite moulds. Cooling rates were measured within situ thermocouples. The conditions needed to stabilize the radial temperature gradient with rotation were established. The microstructure size decreased with increasing rotation, as did the local solidification times. The average grain and dendrite size without imposed rotation is coarser near the mould wall compared with the centre of the casting. This trend is reversed with imposed rotation. Rotation also led to a smaller spread of grain and dendrite size at any chosen height of the casting. These results are discussed in relation to existing theories, and several reasons for an improved heat transfer coefficient with rotation are presented. Forced convective solidification was then carried out for various shapes of integral investment cast Nimonic-90 alloy solidifying under modified conditions that prevented columnar grain formation. Similar results to those recorded for the aluminium case were obtained and are presented here. The major conclusion is that observations indicating a reduction of microstructure spacing during forced convection should also consider improved heat extraction at the mould-metal interface.List of symbols Gr Grashof number =gTZ ^{3 3}/ ³ - g _r acceleration in radial direction - g acceleration in direction - g _z acceleration inZ direction (gravity) - h heat transfer coefficient - k _l thermal conductivity of liquid - Nu _z Nusselt number =hZ/k _l - Pr Prandtl number =/ - Ra Rayleigh numberGr Pr - R radius of mould - Re _r Reynolds number =V ₀ R/ - T temperature - T temperature difference in radial direction - Ta Taylor number = ²4H ⁴ W ²/ ² - V velocity - W r.p.m. - thermal diffusivity - coefficient of volume expansion - viscosity - density Mr G. S. Reddy is also a post graduate student registered at the Banaras Hindu University, Varanasi, India.