Attempted experimental modification of the postlaminectomy membrane by local instillation of recombinant tissue-plasminogen activator gel

A prospective, randomized, blinded trial involving 25 adult mongrel dogs was performed to evaluate whether placement of a fat graft or local instillation of recombinant tissue-plasminogen activator gel (rt-PA) could modify the development of a postlaminectomy membrane. One component of the study involved the performance of a lumbar laminectomy and placement of gel rt-PA, free fat, or no tissue placement over the exposed dura mater. Three months later the animals were killed and sections of spine were removed en bloc, decalcified, and examined histologically. No significant differences were found in the degree of cellular fibrosis or heavy collagen production. A similar laminectomy at another lumbar level was also followed by gel rt-PA, free fat, or no tissue placement. Three months after surgery, surgery was performed again at this level immediately before death. There were no differences in the adhesiveness of the laminectomy membrane to dura mater or roots. It was concluded that the local instillation of gel rt-PA after laminectomy did not inhibit scar formation or scar adherence to the dura mater.     

Early hemodynamic changes at the microcirculatory level and effects of mannitol following focal cryogenic injury

Changes in cerebral blood flow due to infusion of hyperosmolar solutions are of considerable importance in states of raised intracranial pressure. The present study was aimed to evaluate the effects of mannitol on the cerebral microcirculation, in a model of vasogenic brain edema. A right fronto-parietal craniotomy was performed in 30 adult Sprague-Dawley rats. Vasogenic edema was produced by placing dry-ice over the dura for 1 min. The cortical blood flow was monitored for 120 min using a laser-Doppler flowmeter (Perimed, Stockholm, Sweden), and graphics were recorded using a personal computer. Animals were randomly divided into three groups: group 1 (control group) received no mannitol; group 2 was treated with a bolus injection of 20% mannitol (1 mg kg-1); group 3 received the same dose over a 30 min infusion. Mean blood pressure, temperature, and respiratory rate were continuously monitored. At the end of the procedure, an intravenous injection of Evan's blue 2% was given. Results were compared by using repeated measures of analysis of variance and a two-sample t-test at each time. After the production of a cryogenic injury, we found a marked decrease in the cerebral blood flow, whereas mannitol partially reversed that effect. There was not significant difference between groups 2 and 3; however, there was a significant difference between mannitol and control groups after 15 min. During the early phase of vasogenic edema, early use of mannitol did not increase the blood flow, but stabilized it, preventing further decrease. Laser-Doppler flowmetry is a valuable method for continuous estimation of hemodynamic changes in the cerebral microcirculation.     

Role of erythrocytes in leukocyte-endothelial interactions: mathematical model and experimental validation

The binding of circulating cells to the vascular wall is a central process in inflammation, metastasis, and therapeutic cell delivery. Previous in vitro studies have identified the adhesion molecules on various circulating cells and the endothelium that govern the process under static conditions. Other studies have attempted to simulate in vivo conditions by subjecting adherent cells to shear stress as they interact with the endothelial cells in vitro. These experiments are generally performed with the cells suspended in Newtonian solutions. However, in vivo conditions are more complex because of the non-Newtonian flow of blood, which is a suspension consisting of 20-40% erythrocytes by volume. The forces imparted by the erythrocytes in the flow can contribute to the process of cell adhesion. A number of experimental and theoretical studies have suggested that the rheology of blood can influence the binding of circulating leukocytes by increasing the normal and axial forces on leukocytes or the frequency of their collision with the vessel wall, but there have been no systematic investigations of these phenomena to date. The present study quantifies the contribution of red blood cells (RBCs) in cell capture and adhesion to endothelial monolayers using a combination of mathematical modeling and in vitro studies. Mathematical modeling of the flow experiments suggested a physical mechanism involving RBC-induced leukocyte dispersion and/or increased normal adhesive contact. Flow chamber studies performed with and without RBCs in the suspending medium showed increases in wall collision and binding frequencies, and a decrease in rolling velocity in the presence of erythrocytes. Increased fluid viscosity alone did not influence the binding frequency, and the differences could not be attributed to large near-wall excesses of the lymphocytes. The results indicate that RBCs aid in the transport and initial engagement of lymphocytes to the vascular wall, modifying the existing paradigm for immune cell surveillance of the vascular endothelium by adding the erythrocyte as an essential contributor to this process.     

The inhibitory effect of tenidap on leukocyte-endothelial cell adhesion

Treatment of symptomatic unilateral vocal cord paralysis is most frequently surgical. Medialization of the vocal cord using Teflon injection has proved effective; however, studies have shown this technique to produce stiffness of the vocal fold with loss of the "mucosal wave" and concomitantly poor vocal function. As well, overcorrection may occur and is not reversible. Isshiki type 1 medialization thyroplasty has been shown to produce a substantial improvement in vocal quality, as well as preserve the mucosal wave. A number of problems encountered during the performance of Isshiki type 1 thyroplasty has led us to modify the original technique. We have developed a new implant that allows for precise, easily adjustable control of vocal cord medialization. To evaluate the degree of vocal cord medialization afforded by this implant, larynges of fresh male and female cadavers were used as an experimental model. In both larynges, vocal cord medialization was shown to occur in a predictable fashion for the anterior, middle, and posterior segments, as well as in the functionally important inter-arytenoid region. We believe the use of this implant in medialization thyroplasty will allow precise, atraumatic medialization of the paralyzed vocal cord. This greater control over positioning and ease of adjustment should contribute to enhanced vocal quality.     

Effect of stimulation on the stabilization of platelet-fibrinogen interactions

The interaction between fibrinogen and stimulated platelets is a multiphasic process that culminates in the stabilization of ligand binding and reduced accessibility of bound fibrinogen to exogenous antibody. The present study was designed to further explore platelet-fibrinogen interactions by examining the effect of agonist on bound fibrinogen expression and interaction with stimulated platelets as a function of time after ligand binding. Two agents were identified, Zn2+ and phorbol myristate acetate (PMA), which support progressive decreases in bound fibrinogen expression on platelets, but fail to support the stabilization of fibrinogen binding. Sixty min after binding to platelets, approximately 80% of bound fibrinogen remained reversibly associated with Zn(2+)- or PMA-treated platelets and failed to associate with the Triton X-100 insoluble cytoskeleton. In contrast, polyclonal anti-fibrinogen antibody binding decreased by more than 66%. Over the same time course, fibrinogen binding to control platelets, stimulated with thrombin or ADP, was not only accompanied by a 70% decrease in antifibrinogen antibody binding, but also an inability of EDTA or excess exogenous fibrinogen to dissociate more than half of platelet-associated fibrinogen, as well as the progressive association of bound fibrinogen with the platelet cytoskeleton. Costimulation of platelets with ZnCl2 and thrombin or ZnCl2 and ADP enhanced overall fibrinogen binding but not the EDTA-resistant component, and prevented the recovery of irreversibly bound fibrinogen with the Triton X-100 insoluble cytoskeleton. Costimulation of PMA- or Zn(2+)-treated platelets with low doses of A23187, however, restored the stabilization of platelet-fibrinogen interactions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of surgical panel composition on patient outcome at a level I trauma center

OBJECTIVE: To compare the effect of staffing with general surgeons vs trauma specialists on patient outcome at a trauma center. DESIGN: The care of injured patients at a level I urban trauma center serving a population of 2.5 million was the responsibility of 12 surgeons (10 general surgeons and 2 trauma specialists) between January 1 and June 30, 1996 (group 1). Between July 1 and December 31, 1996 (group 2), trauma was the responsibility solely of 4 trauma specialists. An additional comparison was made with those patients in group 1 who were admitted to the general surgeons (group 1A). The outcomes and quality of care for these periods, as determined by the quality assurance screens, were retrospectively analyzed and compared. SETTING: Urban, tertiary care, level I trauma center. PARTICIPANTS: Each trauma and burn patient admitted during the study periods is included in this study. Upon the patient's discharge from the hospital, specially trained nurses completed a review of the patient's stay and entered it into the TraumaOne database (Lancet Technology Inc, Cambridge, Mass). There were 693 trauma patients in group 1 (472 in group 1A) and 734 patients in group 2. MAIN OUTCOME MEASURES: Mortality, length of stay, and 16 quality assurance screens were quantified and compared using chi(2) analyses and t tests. RESULTS: The age and sex of the 2 groups were similar. The mortality rate was 6.2% (43/693) in group 1, 6.1% (29/472) in group 1A, and 6.5% (48/734) in group 2 (P = .80 and P = .78, respectively). When stratified by injury severity score (ISS), lengths of stay were statistically similar, except for patients with an ISS of 0 to 7. Patients with an ISS of 0 to 7 in groups 1 and 1A stayed a mean of 2.6 days, compared with 3.2 days for group 2 (P = .01 and P = .02, respectively). The results of quality assurance screens (missed injury, wound infection, readmission, and 13 others) were similar in the 2 groups. CONCLUSIONS: Transitions in staffing afforded the opportunity to examine patient outcomes by surgeon specialization and frequency of call. In our sample, 12 well-trained surgeons taking call less frequently managed a trauma service as efficiently as a group of 4 trauma specialists, without any differences in morbidity and mortality.     

Effect of cytokines on tumour cell-endothelial interactions

The adherence of tumour cells to microvascular endothelium is believed to be a necessary step in their migration to sites of metastasis. It has been proposed that this process occurs when cell surface molecules on tumour cells bind to complementary sites on endothelial cells. The expression of these endothelial-derived cell adhesion molecules appears to be modulated by cytokines, a broad class of protein mediators which play important roles in immune and inflammatory reactions. It has been found by ourselves and others that exposure of endothelium to some cytokines augments the adhesion of inflammatory cells as well as tumour cells in in vitro assays. We used a murine model consisting of P815 mastocytoma cells and microvascular endothelium and found that pretreatment of endothelial monolayers with TNF-alpha, IL-1, LPS or PMA augmented the number of tumour cells that attach in a dose-dependent fashion. FACS analysis showed that the change in binding was due to an increase in the expression of VCAM-1 on the surface of the endothelial cell. Methylxanthines (caffeine and theophylline) as well as "classical" calcium-mobilizing agents (ionomycin and thapsigargin) inhibited the expression of VCAM-1 in MME. We also studied the possible mechanisms of TNF-alpha signal transduction in endothelial cells. We examined the involvement of protein kinases in the TNF-alpha effect. Although we found that inhibitors of PKC could inhibit the TNF-alpha effect, our studies suggest that the "classical" PKC pathway is not completely responsible for signaling since TNF-alpha did not cause translocation of PKC to the cell membrane and its effect could not be completely mimicked by PMA. We also studied the effect of TGF-beta on the binding of tumour cells to endothelium. Exposure of endothelium to TGF-beta led to the inhibition of both basal and TNF-alpha enhanced binding of P815 cells. Inhibitors of G-proteins do not abolish TGF-beta action, and PKC and PKA activators elicit an opposite effect. However, TGF-beta-mediated inhibition of both basal binding and TNF-alpha-enhanced P815 binding to endothelium is completely abolished in the presence of the protein phosphatase inhibitor okadaic acid suggesting that TGF-beta elicits its effect by stimulating protein phosphatase activity.     

Evolutionary conserved rigid module-domain interactions can be detected at the sequence level: the examples of complement and blood coagulation proteases

Several extracellular modular proteins, including proteases of the complement and blood coagulation cascades, are shown here to exhibit conserved sequence patterns specific for a particular module-domain association. This was detected by comparative analysis of sequence variability in different multiple sequence alignments, which provides a new tool to investigate the evolution of modular proteins. A first example deals with the proteins featuring a common complement control protein (CCP) module-serine protease (SP) domain pattern at their C-terminal end, defined here as the CCP-SP sub-family. These proteins include the complement proteases C1r, C1s and MASPs, the Limulus clotting factor C, and the proteins of the haptoglobin family. A second example deals with blood coagulation factors VII, IX and X and protein C, all featuring a common epidermal growth factor (EGF)-SP C-terminal assembly. Highly specific motifs are found at the connection between the CCP or EGF module and the activation peptide of the SP domain: [P/A]-x-C-x-[P/A]-[I/V]-C-G-x-[P/S/K] in the case of the CCP-SP proteins, and C-x-[P/S]-x-x-x-[Y/F]-P-C-G in the case of the EGF-SP proteins. Each motif is strictly conserved in the whole sub-family and it is detected in no more than one other known protein sequence. Strikingly, most of the conserved residues specific to each sub-family appear to be clustered at the interface between the SP domain and the CCP or EGF module. We propose that a rigid module-domain interaction occurs in these proteins and has been conserved through evolution. The functional implications of these assemblies, underlined by such evolutionary constraints, are discussed.     

The microcirculatory bed of the human epididymis

Based on the material of 24 human epididymides at ages 18 to 54, hemomicrocirculatory bed was studied of epididymis in man with the aid of a complex of morphologic techniques (injection of 20% Chinese ink-gelatine suspension, injection of a weak solution of caustic silver, transmission electron microscopy). It has been ascertained that architectonics and ultrastructural features of various links of the hemomicrocirculatory bed have signs of regional specificity for the subcapsular vascular network, small seminal ducts of caput epididymidis, ductus epididymidis of the head, body and tail of the organ. Reasons are discussed why specific hemomicrocirculatory bed should be caused to develop in different parts of the organ.     

Effect of oral contraceptives on the blood lipid level

Secretion of estrogens in the urine and the content of cholesterol and triglycerides in blood plasma were studied in 36 healthy women 24 of whom took oral contraceptives including estrogenic and gestagenic components to prevent conception. It proved that after the first course of medication the overall estrogen excretion reduces at the expense of all fractions but more so at the expense of the active fraction. Beginning with the 2nd week the cholesterol level in the blood plasma increases and the level of triglycerides grows considerably. The increase in these indices was observed in the first 6 months when the oral contraceptives were taken. With the preparations taken for a longer time the blood plasma cholesterol and triglyceride levels became stable.     

Public health at the municipal level

The purposes of health protection and fortification in various sociums and conditions of attaining these purposes may differ at various levels (federal, regional and municipal) of the society organization. At the municipal level the fortification and protection of population health may be regarded as the chief idea, aim, and criterion of the optimal functioning and development of a socium of the given level, reflecting the measure of efficacy of providing the immediate vital activity of the population. The task of social management is as follows: the share of the national output, created by the socium, is to be distributed so that it were possible for this socium to permanently recreate and maintain at a certain qualitative level its labor resources--the major energy potential of the socium, and adequately provide and guarantee human rights as the main value of the society. A logistic model of the management structure ensuring population health at a municipal level is presented. For health control and for attaining this aim it is necessary to transfer the maximum of legal functions to the municipal level of society administration. It is desirable that every citizen, and not only administration, be involved in the reformation of the society as regards health protection, and that the citizens be personally interested in this process.     

Development of a TEM to study in situ structural and chemical changes at an atomic level during gas-solid interactions at elevated temperatures

A Philips (Eindhoven, The Netherlands) 430 (300 keV) high resolution transmission electron microscope has been modified for in situ study of gas-solid interactions at elevated temperatures. This microscope can be best described as a synthesis, processing, and characterization laboratory for nano-size materials. A differentially pumped environmental cell (E-cell), capable of handling up to 20 torr of gas pressure, is fitted in the objective lens pole-piece gap. Single-tilt or double-tilt heating holders can be used to heat the samples up to 1,300 degrees C and 850 degrees C, respectively. The system can handle any non-corrosive gases such as H2, O2, N2, NH3, CO, water vapor. Electron diffraction patterns are used to elucidate the reaction path and to identify stable and/or metastable phases formed. Time, temperature, and pressure resolved electron diffraction patterns can also be used to estimate the thermodynamical conditions for the onset of a reaction and the stability range of different phases observed during the process can also be determined. The high resolution imaging capabilities enable elucidation of the basic structural mechanisms involved at near atomic level. The TV rate camera/video recording system is used to measure the reaction rates (kinetics of the reaction). A post projector energy filter (Gatan Imaging Filter, GIF) is attached at the bottom of the microscope in order to filter the inelastic scattering from the gases/thick samples as well as to obtain energy filtered images (chemical maps). The GIF can also be used as a parallel electron energy loss spectrometer (PEELS) to obtain changes in the sample composition during the reactions. The changes in the near-edge structures of PEELS spectrum is used to monitor changes in bonding and/or chemical environment elements during reaction. The chemical maps obtained can also be used to identify preferred regions of gas reactions, e.g., grain boundaries or surfaces, etc. Various modifications of the microscope are described in detail, with suitable examples showing the performance.     

Phosphorylation of the cytoplasmic domain of E-selectin is regulated during leukocyte-endothelial adhesion

E-selectin, a selectin expressed on activated vascular endothelium, supports rolling and stable adhesion of leukocytes at sites of inflammation. Previously, we have reported that leukocyte adhesion to cultured endothelial cells induces association of the cytoplasmic domain of E-selectin with cytoskeletal elements, suggesting that outside-in signaling may occur during E-selectin-mediated adhesion. To investigate this potential signaling function of E-selectin, HUVEC activated with recombinant human IL-1beta (10 U/ml, 4 h) were labeled with [32P]orthophosphate, and E-selectin was immunoprecipitated using mAb H18/7. Autoradiography revealed constitutive phosphorylation of E-selectin in these cells and time-dependent dephosphorylation following adhesion of HL-60 cells. Cross-linking of cell surface E-selectin using H18/7 and a polyclonal secondary Ab induced E-selectin dephosphorylation, as did adhesion of beads coated with recombinant P-selectin glycoprotein ligand-1 (PSGL-1), an E-selectin ligand. Using adenoviral vector-mediated transfection in HUVEC of a tail-less E-selectin and phosphoamino acid analysis, we documented phosphorylation occurring exclusively within the cytoplasmic domain and involving serine residues. Additional experiments using a series of cytoplasmic domain mutants of E-selectin expressed in COS-7 cells localized the regions that were constitutively phosphorylated. Preincubation with okadaic acid and sodium vanadate abrogated adhesion-induced dephosphorylation of E-selectin. Thus, E-selectin, which is constitutively phosphorylated in cytokine-activated human endothelial cells, undergoes an enzymatically regulated dephosphorylation following leukocyte adhesion. This process appears to be triggered by multivalent ligand binding and/or cross-linking of cell surface E-selectin. Ligand-dependent regulation of the phosphorylation of E-selectin's cytoplasmic domain provides additional evidence for a transmembrane signaling function of this molecule during leukocyte-endothelial interactions.     

Effect of the double-bind communication on the anxiety level of normals

A double-bind communication was broken into its 2 components, punishment and contradictory material, and administered to 4 groups of 39 female college sophomores. There were 2 punishment conditions, 76% variable-interval white noise and no punishment, and 2 levels of stimulus material, contradictory and noncontradictory. These 4 factors were manipulated in a 2 * 2 factorial design over 3 levels of trait anxiety. The criterion measure was change in state anxiety as measured by 5 tests. It was found that the interaction of the 2 components, that is, the full double-bind condition, was significant at the .01 level. The double-bind, as opposed to the component conditions, formed a situation to which the Ss seemed unable to adapt. (17 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

A conformational model for the action of general anesthetics at the membrane level. II. Experimental observations on the effects of anesthetics on lipid fluidity and lipid protein interactions

We have investigated the effect of general anesthetics (the normal alcohol series up to pentanol, halothane, pentrane, ether, chloroform, and ketamine) on lipid fluidity of phospholipid vesicles and mitochondrial and erythrocyte membranes by using spin labels and fluorescent probes. The spin labels used (5- and 16-doxyl stearic acids) show that all anesthetics tested have a slight fluidizing effect on lipid vesicles but induce a very strong increase in mobility of spin labels in mitochondria and lower in erythrocyte ghosts. These results are interpreted as a labilization of lipid protein interactions at all depths in the bilayer. The fluorescent molecules ANS and NPN, which probe the glycerol region and the core of the bilayer respectively, show a decrease of fluorescence induced by alcohols, halothane, ether, chloroform in both lipid vesicles and membranes. The decrease of fluorescence is due to decreased quantum yield as shown by double reciprocal plots of probe fluorescence against membrane concentration. The fluorescence decrease is interpreted mainly as an increase in fluidity of the lipid bilayer and not as an increase of polarity of the probe environment. The effect of ketamine is that of fluidization in the bilayer core (NPN) but of increased rigidity in the glycerol region (ANS) perhaps due to the amphipathic character of this anesthetic, that is supposed to bind in the polar region of the bilayer. Pentrane also induces fluidization in the bilayer core (NPN) but has a peculiar effect near the surface (ANS): in lipid vesicles it induces a fluorescence decrease, whereas an increase is seen in mitochondrial membranes. These complex effects are considered as the result of some specific change in the lipid protein interactions in the region probed by ANS. The effects of anesthetics on maximal NPN fluorescence (Fo) have been usually found to be stronger in mitochondrial membranes than in lipid vesicles, thus confirming the results of the spin label studies, showing a labilization of lipid protein interactions induced by anesthetics. The effects on Fo of ANS, however, appear to be stronger in lipid vesicles than in membranes. These findings indicate that the presence of the proteins counteracts the perturbation induced by anesthetics at the level of the membrane surface, in contrast with the disruption of lipid protein interactions observed in the membrane hydrophobic areas.     

Mirror symmetry breaking at the molecular level

Reasoning from two basic principles of molecular physics, P invariance of electromagnetic interaction and the second law of thermodynamics, one would conclude that mirror symmetry retained in the world of chiral molecules. This inference is fully consistent with what is observed in inorganic nature. However, in the bioorganic world, the reverse is true. Mirror symmetry there is definitely broken. Is it possible to account for this phenomenon without going beyond conventional concepts of the kinetics of enantioselective processes? This study is an attempt to survey all existing hypotheses containing this phenomenon.