Ab initio crystallographic structure determination of insulin from protein to electron density without crystal handling.

Insulin crystals suitable for cryogenic data collection and structure determination by single-wavelength anomalous scattering (SAS) were obtained by a self-optimization screening process in a single capillary tube without manipulation of the crystals at any time. Using the counter-diffusion crystallization technique, screening for optimal conditions for crystal growth, incorporation of a strong anomalous scattering halide and cryogenic solution took place simultaneously in a single capillary tube. The crystals in the capillaries can be placed directly in the cryostream for data collection using a conventional home-laboratory X-ray source.

Prolactin is a component of the human synovial liquid and modulates the growth and chondrogenic differentiation of bone marrow-derived mesenchymal stem cells.

The hormone prolactin (PRL) is the product of a single gene synthesized by pituitary and many extrapituitary tissues. In this study, we have purified and sequenced by mass spectrometry a 29 kDa protein from human synovial liquid, bound to the proteoglycan component of synovial liquid that showed an identical sequence in 20 amino acids to hPRL. We have also found PRL receptor (PRLR) in human knee tissues.

Are myeloma patients with renal failure candidates for autologous stem cell transplantation?

Introduction: Renal function is one of the most important prognostic factors in multiple myeloma (MM). Patients with renal failure are generally excluded from high dose therapy even though they display a poor prognosis with conventional chemotherapy schemes. The aim of this study was to analyze the outcome of MM patients with renal insufficiency undergoing autologous stem cell transplantation (ASCT), including the evaluation of the quality of PB stem cell collections, kinetics of engraftment, transplant-related mortality, response to high dose chemotherapy and survival.

Ovotransferrin is a matrix protein of the hen eggshell membranes and basal calcified layer.

The eggshell is an highly ordered structure deposited in the distal oviduct and composed of calcium carbonate and an organic matrix which is believed to influence its fabric. We have identified ovotransferrin as an 80 kDa matrix protein observed at high concentration in the uterine fluid at the initial stage of shell mineralization, by N-terminal sequencing and western blotting using monoclonal and polyclonal antibodies. It is present in extracts from demineralized eggshell and was localized by immunofluorescence in the eggshell membranes and mammillae, which are the sites of calcite nucleation.

Myeloablative treatments for multiple myeloma: update of a comparative study of different regimens used in patients from the Spanish registry for transplantation in multiple myeloma.

After a previous analysis that did not detect clear differences in the results of three conditioning regimens used for autologous stem cell transplantation (ASCT) in patients from the Spanish Registry for Transplant in Multiple Myeloma (MM), we have updated the registry, including a larger number of cases and a fourth conditioning regimen with a longer follow-up. We evaluate 472 MM patients treated with 200 mg/m2 melphalan (MEL200), 135 patients treated with 140 mg/m2 melphalan plus total body irradiation [(MEL140 + TBI)], 186 patients treated with 12 mg/kg busulphan plus 140 mg/m2 melphalan (BUMEL) and 28 patients treated with 14 mg/kg busulphan followed by cyclophosphamide 120 mg/kg (BUCY). There were no significant differences in respect to either transplant related death or haematological recovery, regardless of growth factor use, between the four conditioning programs.

Soaking: the effect of osmotic shock on tetragonal lysozyme crystals.

Protein crystals crack when they are soaked in a solution with ionic strength sufficiently different from the environment in which they grew. It is demonstrated for the case of tetragonal lysozyme that the forces involved and the mechanisms that lead to the formation of cracks are different for hypertonic and hypotonic soaking. Tetragonal lysozyme crystals are very sensitive to hypotonic shocks and, after a certain waiting time, cracks always appear with a characteristic pattern perpendicular to the crystallographic c axis.

Polarization-dependent XANES study of Bi2Sr2Ca(1-x)Pr(x)Cu2O8-delta insulating single crystal.

Pr doping has been extensively studied in cuprate superconductors to understand the mechanism of quenching of superconductivity in cuprate perovskites. Experience has revealed that it acts differently in different cuprate perovskites. We have made high resolution polarization XANES measurements on a Pr-doped single crystal within the ab-plane on the Cu K and Pr L III absorption edges to ascertain the valence state of Pr cation and thereby try learning how it is quenching superconductivity in the present case.

O K and Cu LIII edge study of itinerant holes in I2-, Hgl2- and HgBr2- intercalated BSCCO(2212) single crystals.

Intercalation effect on BSCCO (2212) system, although it increases the interlayer distance and the c-axis remarkably, produces only a small change in the transition temperature. Thus, amongst other things, intercalation provides an effective method to investigate the influence of the interblock coupling. Electrons are transferred from the host Cu-O2 layers to the guest molecules I2, HgBr2, HgI2 leading to evolution of the Tc.

Structure of tetragonal hen egg-white lysozyme at 0.94 A from crystals grown by the counter-diffusion method.

Very high quality crystals of tetragonal hen egg-white lysozyme were grown in the Advanced Protein Crystallization Facility (APCF) on board the Space Shuttle using a modified free-interface diffusion (FID) reactor designed ad hoc to have a longer diffusion path. This design allows the performance of true counter-diffusion experiments. Crystals were obtained under the classical chemical conditions defined 50 y ago with NaCl as a crystallizing agent and acetate pH 4.

Experimental evidence for the stability of the depletion zone around a growing protein crystal under microgravity.

Experimental evidence is presented for the first time for the development and time evolution of concentration-depletion zones around protein crystals growing in microgravity and gelled on-ground experiments. Crystal motion and buoyancy-driven fluid movements as a result of residual accelerations and g-jitters are demonstrated to have an adverse effect on the stability of these depletion zones, provoking the breakdown of their radial symmetry. These findings may explain some of the controversial results reported on the quality of single crystals grown under microgravity in previous space missions.

Identification and localization of lysozyme as a component of eggshell membranes and eggshell matrix.

The avian eggshell is a composite biomaterial composed of non-calcifying eggshell membranes and the overlying calcified shell matrix. The calcified shell forms in a uterine fluid where the concentration of different protein species varies between the initial, rapid calcification and terminal phases of eggshell deposition. The role of these avian eggshell matrix proteins during shell formation is poorly understood.

The effect of avian uterine fluid on the growth behavior of calcite crystals.

Eggshell formation takes place on the eggshell membrane in an acellular medium, the uterine fluid that contains the inorganic minerals and precursors of the organic matrix. The high degree of eggshell structure could be due to an interaction between calcium carbonate and the organic matrix. The aim of this study was to demonstrate such an interaction by measuring the effect in vitro of uterine fluid collected at various phases of shell formation on precipitation kinetics, size, and morphology of calcite crystals.

Remission status defined by immunofixation vs. electrophoresis after autologous transplantation has a major impact on the outcome of multiple myeloma patients.

We have retrospectively analysed 344 multiple myeloma (MM) patients (202 de novo patients) treated in a non-uniform way in whom high-dose therapy and autologous stem cell transplantation (ASCT) response was simultaneously measured by both electrophoresis (EP) and immunofixation (IF). Patients in complete remission (CR) by EP were further subclassified as CR1 when IF was negative and CR2 when it remained positive. Partial responders (PR) were also subclassified as PR1 (very good PR, > 90% reduction in M-component) or PR2 (50-90% reduction).

Conditioning regimens in autologous stem cell transplantation for multiple myeloma: a comparative study of efficacy and toxicity from the Spanish Registry for Transplantation in Multiple Myeloma.

High-dose chemoradiotherapy conditioning regimens for autologous stem cell transplantation (ASCT) are generally held to give similar results in multiple myeloma (MM), but no specific comparative study has been published. We addressed this issue by comparing the main high-dose chemoradiotherapy regimens used in the Spanish Registry. Patient cohorts included 315 cases treated with 200 mg/m2 melphalan (MEL200), 127 patients with 140 mg/m2 melphalan plus total body irradiation (MEL140 + TBI) and 121 cases with 12 mg/kg busulphan plus 140 mg/m2 melphalan (BUMEL).

Transgenic mice expressing bovine GH develop arthritic disorder and self-antibodies.

We observed disability of movement in 6-month-old transgenic mice expressing the fusion gene coding for the bovine GH (bGH) under the transcriptional control of phosphoenolpyruvate carboxykinase promoter (PEPCK-bGH). Histological study of the knee joint showed altered synovial and tibial articular cartilage tissues. In the cartilage the following observations were made: (i) generalized loss of the normal zonal structure and presence of clefts, and (ii) profound alterations in chondrocyte growth/differentiation processes consistent with hypertrophy.

Prolactin (PRL)-PRL receptor system increases cell proliferation involving JNK (c-Jun amino terminal kinase) and AP-1 activation: inhibition by glucocorticoids.

PRL receptor (PRLR) signal transduction supports PRL-induced growth/differentiation processes. While PRL is known to activate Jak2-Stat5 (signal transducer and activator of transcription 5) signaling pathway, the mechanism by which cell proliferation is stimulated is less known. We show that PRL induces proliferation of bovine mammary gland epithelial cells and AP-1 site activation.

In-situ measurement of rocking curves during lysozyme crystal growth.

The rocking curve of protein crystals contains a lot of useful information concerning crystal quality, most of which is lost owing to the superimposition of spurious features appearing in these fragile materials after growth, during handling and mounting. To minimize such data spoiling, an experimental setup to perform in situ X-ray diffraction experiments during crystal growth has been designed. The setup, which includes video observation to allow the correlation of crystal shape, size and growth rate with X-ray data, has been used to assess the mosaicity of tetragonal lysozyme crystals during crystal growth.

Mosaic spread characterization of microgravity-grown tetragonal lysozyme single crystals.

Mosaic spread values for crystals grown in microgravity were measured using synchrotron radiation. Full width at half maximum (FWHM) values for diffraction line profiles in the range 10-20" (arc seconds, 1" = 1 degrees /3600) were observed. These values are similar to those measured for crystals grown on earth using the gel-acupuncture method.

Detection of antibodies to Candida albicans germ tubes for diagnosis and therapeutic monitoring of invasive candidiasis in patients with hematologic malignancies.

We prospectively investigated the ability of detection of antibodies to Candida albicans germ tubes (CAGT) to diagnose invasive candidiasis in 95 consecutive admissions of 73 patients with hematologic disorders undergoing intensive chemotherapy. The episodes were divided into three groups according to clinical and microbiological diagnosis. Group 1 comprised eight admissions of eight patients with invasive candidiasis.

The short form of the prolactin (PRL) receptor silences PRL induction of the beta-casein gene promoter.

The PRL receptor (PRLR) is a member of the cytokine receptor superfamily. Rats and mice express two forms of PRLR, short (SPRLR) and long (LPRLR), which differ in the length and sequence of their cytoplasmic domains. We have analyzed the ability of each form of rat PRLR to transduce lactogenic signals in a bovine mammary gland epithelial cell line.

RESEARCH: Managing Mountainous Degraded Landscapes After Farmland Abandonment in the Central Spanish Pyrenees.

/ Plant succession and pasture resources have been studied in abandoned fields of the central Spanish Pyrenees, in an environment severely affected by strong demographic pressure in the past. Several hydromorphological features (runoff and sediment yield) were also analyzed for different environments of the abandoned fields, in order to forecast the effects of their reclamation and transformation into areas for livestock use. The availability and accessibility of pastures as well as soil and water conservation is related to the process of colonization of Genista scorpius.

Prolactin receptor is associated with c-src kinase in rat liver.

The mechanism of action of the pituitary hormone PRL was studied in hepatocytes of lactating rats. PRL receptor immune complexes obtained from liver lysates have an associated tyrosine kinase activity. The tyrosine kinase has been identified in isolated hepatocytes as pp60c-src.