Fewer bone disease events, improvement in bone remodeling, and evidence of bone healing with bortezomib plus melphalan-prednisone vs. melphalan-prednisone in the phase III VISTA trial in multiple myeloma

Objectives: Bone disease is a key presenting feature of myeloma. This post hoc analysis of the phase III VISTA trial of bortezomib plus melphalan–prednisone (VMP) vs. MP in previously untreated myeloma patients assessed clinical bone disease events and changes in alkaline phosphatase (ALP), a marker for osteoblast activation, and serum Dickkopf‐1 (DKK‐1), an inhibitor of osteoblast differentiation, during treatment. Methods: Patients received nine 6‐wk cycles of VMP (bortezomib 1.3 mg/m², days 1, 4, 8, 11, 22, 25, 29, 32, cycles 1–4, days 1, 8, 22, 29, cycles 5–9, plus melphalan 9 mg/m² and prednisone 60 mg/m², days 1–4, cycles 1–9; N = 344) or MP alone (N = 338). Results: Rates of bisphosphonates use during treatment (73% vs. 82%), progression because of worsening bone disease (3% vs. 11%), and requirement for subsequent radiotherapy (3% vs. 8%) were lower with VMP vs. MP. Median maximum ALP increase was significantly higher with VMP vs. MP overall (49.7% vs. 30.3%, P = 0.029), and higher by response group (complete response [CR]: 68.7% vs. 43.9%; partial response [PR]: 41.5% vs. 31.2%). Greater maximum ALP increase was strongly associated with achievement of CR (P ≤ 0.0001) and CR/PR (P ≤ 0.01). Median DKK‐1 decreased with VMP by 694.4 pg/mL and increased with MP by 1273.3 pg/mL from baseline to day 4 (P = 0.0069). Available radiologic data revealed evidence of bone healing in 6/11 VMP‐treated patients, who achieved best responses of three CR, one PR, and two stable disease. Conclusions: These results suggest a positive effect of bortezomib on bone metabolism and potentially bone healing in myeloma.     

Addictive behavior among young people in Ukraine: a pilot study

The AUDIT-like tests system was created for complex assessment and evaluation of the addictive status of adolescents in a Ukrainian population. The AUDIT-like tests system has been created from the Alcohol Use Disorders Identification Test (AUDIT) developed by the World Health Organization. The AUDIT-like tests were minimally modified from the original AUDIT. Attention was brought to similarities between stages of different addictions (TV, computer games, the Internet, etc.) and alcohol addiction. Seventeen AUDIT-like tests were created to detect the different types of chemical and non-chemical addictions.     

Determination of human leukocytic interferon by a microfluorometric immunological method

The possibility of quantitative determination of human leukocytic interferon by means of anti-interferon antibodies labeled with fluorescein isothiocyanate (FITC) was studied. As a result of prolonged immunization of a donkey with a preparation of human leukocytic interferon and subsequent fractionation and immunosorption, a donkey antiinterferon immunoglobulin with high specific activity was obtained. This immunoglobulin was labeled with FITC and used for quantitative determination of human leukocytic interferon in direct and indirect immunologic inhibition of fluorescence tests. The titers of the various preparations of human leukocytic interferon in the inhibition of fluorescence test were comparable with the titers of these same preparations in the test of inhibition of virus cytopathic action by interferon.Laboratory of Immunology of Leukemias, Central Institute of Hematology and Blood Transfusion, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Fedorov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 87, No. 4, pp. 322–325, April, 1979.     

General anesthetics inhibit gap junction communication in cultured organotypic hippocampal slices

Gap junctions are protein channels that directly connect the cytosol of neighboring cells, thus forming electrical synapses and promoting synchronous neuronal activities. Such activities lead to the initiation and propagation of electroencephalogram oscillations implicated in cognition and consciousness. In this study, we investigated the effects of propofol, thiopental, and halothane on gap junction communication in cultured organotypic hippocampal slices by recovery of fluorescence after photo bleaching (FRAP) technique and electrophysiological recordings. Propofol 15 microM and thiopental 10 microM attenuated gap junction communication in slice cultures by 46.7% +/- 4.5% and 48.8% +/- 5.5%, respectively, as measured by FRAP. Smaller concentrations of propofol 5 microM and thiopental 2 microM did not change gap junction coupling. Accompanying the decreased gap junction communication, hippocampus slice cultures exposed to propofol 15 microM and thiopental 10 microM were found to have reduced electrophysiologic spontaneous discharges and primary after discharges evoked by a tetanic train of 50 Hz for 2 s. On the other hand, halothane 0.64 mM, a concentration slightly larger than twice its minimum alveolar concentration had no effect on gap junction coupling while halothane 2.8 mM blocked FRAP by 70%. The current study illustrates that anesthetic concentrations of propofol and thiopental, but not halothane, attenuate gap junction communication in cultured hippocampal slices. Suppression of gap junction function could compound the mechanisms of anesthetic actions.     

Visualization of cytoplasmic diffusion within living myelin sheaths of CNS white matter axons using microinjection of the fluorescent dye Lucifer Yellow

The compactness of myelin allows for efficient insulation defining rapid propagation of action potentials, but also raises questions about how cytoplasmic access to its membranes is achieved, which is critical for physiological activity. Understanding the organization of cytoplasmic ('water') spaces of myelin is also important for diffusion MRI studies of CNS white matter. Using longitudinal slices of mature rat spinal cord, we monitored the diffusion of the water-soluble fluorescent dye Lucifer Yellow injected into individual oligodendrocytes or internodal myelin. We show that living myelin sheaths on CNS axons are fenestrated by a network of diffusionally interconnected cytoplasmic 'pockets' (1.9 ± 0.2 pockets per 10μm sheath length, n=58) that included Schmidt-Lanterman clefts (SLCs) and numerous smaller compartments. 3-D reconstructions of these cytoplasmic networks show that the outer cytoplasmic layer of CNS myelin is cylindrically 'encuffing', which differs from EM studies using fixed tissue. SLCs were found in different 'open states' and remained stable within a 1-2hour observation period. Unlike the peripheral nervous system, where similarly small (<500Da) molecules diffuse along the whole myelin segment within a few minutes, in mature CNS this takes more than one hour. The slower cytoplasmic diffusion in CNS myelin possibly contributes to its known vulnerability to injury and limited capacity for repair. Our findings point to an elaborate cytoplasmic access to compact CNS myelin. These results could be of relevance to MRI studies of CNS white matter and to CNS repair/regeneration strategies.     

Protein tyrosine phosphatase kappa and SHP-1 are involved in the regulation of cell-cell contacts at adherens junctions in the exocrine pancreas

BACKGROUND: We have previously shown that cell contacts between pancreatic acinar cells dissociate early in pancreatitis and that this is a prerequisite for the development of pancreatic oedema. Here we studied the underlying mechanism. METHODS: Employing experimental caerulein induced pancreatitis in vivo and isolated pancreatic acini ex vivo, in conjunction with protein chemistry, morphology, and electron microscopy, we determined whether cell contact regulation in the pancreas requires or involves: (1) changes in cadherin-catenin protein expression, (2) tyrosine phosphorylation of adhesion proteins, or (3) alterations in the actin cytoskeleton. RESULTS: During initial cell-cell contact dissociation at adherens junctions, expression of adhesion proteins remained stable. At time points of dissociated adherens junctions, the cadherin-catenin complex was found to be tyrosine phosphorylated and internalised. The receptor type protein tyrosine phosphatase (PTP)kappa was constitutively associated with the cadherin-catenin complex at intact cell contacts whereas following the dissociation of adherens junctions, the internalised components of the cadherin-catenin complex were tyrosine phosphorylated and associated with the cytosolic PTP SHP-1. In isolated acini, inhibition of endogenous protein tyrosine phosphatases alone was sufficient to induce dissociation of adherens junctions analogous to that found with supramaximal caerulein stimulation. Dissociation of actin microfilaments had no effect on adherens junction integrity. CONCLUSIONS: These data identify tyrosine phosphorylation as the key regulator for cell contacts at adherens junctions and suggest a definitive role for the protein tyrosine phosphatases PTPkappa and SHP-1 in the regulation, maintenance, and restitution of cell adhesions in a complex epithelial organ such as the pancreas.     

CD40L blockade prevents autoimmune encephalomyelitis and hampers TH1 but not TH2 pathway of T cell differentiation

 Although it is well established that CD40 and its ligand (CD40L) play pivotal roles in the development of humoral immunity, their roles in cell-mediated immunity and cell-mediated autoimmune diseases are not well defined. We report here that CD40:CD40L interaction is crucial for the development of experimental autoimmune encephalomyelitis (EAE), a prototype TH1-cell mediated autoimmune disease. Specific blockade of CD40L at the time of immunization markedly suppressed the incidence, mortality, day of onset, and clinical scores of EAE in (PLJ×SJL) F1 mice. Importantly, the disease suppression was not associated with anergy or deletion of autoreactive T cells but was accompanied by a drastic alteration of their cytokine profiles. The production of interferon (IFN)-γ was markedly suppressed while that of interleukin (IL)-4 enhanced. These results suggest that CD40:CD40L interaction plays important roles in the differentiation of autoreactive TH1 versus TH2 cells in vivo, and that CD40L blockade is effective in preventing autoimmune encephalomyelitis. Received: 7 November 1996 / Accepted: 3 April 1997     

Properties of WC–10%Co–4%Cr Detonation Spray Coating Deposited on the Al–4%Cu–1%Mg Alloy

One of the methods of local improvement of the wear resistance of aluminum alloy parts is the deposition of hard tungsten carbide-based coatings on the surfaces subjected to intense external influence. This paper is devoted to the characterization of the WC–10Co–4Cr (wt.%) coating deposited on an Al–4Cu–1Mg (wt.%) alloy by the detonation spray method. In comparison with the common thermal spray techniques like High Velocity Oxygen Fuel (HVOF) or Atmospheric Plasma Spraying (APS), the heat input delivered to the substrate during detonation spray is significantly lower, that is especially important in case of coating deposition on aluminum alloys. The paper presents the results of morphology investigation, microstructure, phase composition, microhardness, and cohesive strength of deposited carbide-based detonation spray coating. Results showed that the coating has a porosity less than 0.5% and the carbide grain refinement down to the submicron size during coating deposition was detected. According to the investigation, the variation of spraying distance from 270 to 230 mm does not influence on the coating microstructure and composition.