From CT scanning to 3D printing technology: a new method for the preoperative planning of a transcutaneous bone-conduction hearing device.

SUMMARY: The aim of the present study was to assess the feasibility and utility of 3D printing technology in surgical planning of a transcutaneous bone-conduction hearing device (Bonebridge®) (BB), focusing on the identification of the proper location and placement of the transducer. 3D printed (3DP) models of three human cadaveric temporal bones, previously submitted to CT scan, were created with the representation of a topographic bone thickness map and the sinus pathway on the outer surface. The 3DP model was used to detect the most suitable location for the BB. A 3DP transparent mask that faithfully reproduced the surface of both the temporal bone and the 3DP model was also developed to correctly transfer the designated BB area. The accuracy of the procedure was verified by CT scan: a radiological marker was used to evaluate the degree of correspondence of the transducer site between the 3DP model and the human temporal bone. The BB positioning was successfully performed on all human temporal bones, with no difficulties in finding the proper location of the transducer. A mean error of 0.13 mm was found when the transducer site of the 3DP model was compared to that of the human temporal bone. The employment of 3D printing technology in surgical planning of BB positioning showed feasible results. Further studies will be required to evaluate its clinical applicability.

Early-stage roughening of the polymer-polymer interface approaching the glass transition temperature by real-time neutron reflection.

The early-stage roughening of the interface between thin deuterated poly(methyl methacrylate) (d-PMMA) layers on thick polystyrene (PS) films was studied as a function of the temperature using real-time specular neutron reflectivity. By measuring the growth of the interface roughness as a precursor of the dewetting, the characteristic time constant of the early stages of the process was studied as a function of the temperature approaching the glass transition temperature (T(g)) of the two polymers from above and compared with the prediction of the growth of the interface by the spinodal process. Both solid and liquid regimes were probed, in which the viscosity of the thin film or the substrate dominates respectively. The characteristic growth time of the process also depends on the upper film thickness to a power of 5 or 6 in the solid or liquid regimes, respectively, as predicted by the theory of spinodal dewetting.

Steroid hormones and neurosteroids in normal and pathological aging of the nervous system.

Without medical progress, dementing diseases such as Alzheimer's disease will become one of the main causes of disability. Preventing or delaying them has thus become a real challenge for biomedical research. Steroids offer interesting therapeutical opportunities for promoting successful aging because of their pleiotropic effects in the nervous system: they regulate main neurotransmitter systems, promote the viability of neurons, play an important role in myelination and influence cognitive processes, in particular learning and memory. Preclinical research has provided evidence that the normally aging nervous system maintains some capacity for regeneration and that age-dependent changes in the nervous system and cognitive dysfunctions can be reversed to some extent by the administration of steroids. The aging nervous system also remains sensitive to the neuroprotective effects of steroids. In contrast to the large number of studies documenting beneficial effects of steroids on the nervous system in young and aged animals, the results from hormone replacement studies in the elderly are so far not conclusive. There is also little information concerning changes of steroid levels in the aging human brain. As steroids present in nervous tissues originate from the endocrine glands (steroid hormones) and from local synthesis (neurosteroids), changes in blood levels of steroids with age do not necessarily reflect changes in their brain levels. There is indeed strong evidence that neurosteroids are also synthesized in human brain and peripheral nerves. The development of a very sensitive and precise method for the analysis of steroids by gas chromatography/mass spectrometry (GC/MS) offers new possibilities for the study of neurosteroids. The concentrations of a range of neurosteroids have recently been measured in various brain regions of aged Alzheimer's disease patients and aged non-demented controls by GC/MS, providing reference values. In Alzheimer's patients, there was a general trend toward lower levels of neurosteroids in different brain regions, and neurosteroid levels were negatively correlated with two biochemical markers of Alzheimer's disease, the phosphorylated tau protein and the beta-amyloid peptides. The metabolism of dehydroepiandrosterone has also been analyzed for the first time in the aging brain from Alzheimer patients and non-demented controls. The conversion of dehydroepiandrosterone to Delta5-androstene-3beta,17beta-diol and to 7alpha-OH-dehydroepiandrosterone occurred in frontal cortex, hippocampus, amygdala, cerebellum and striatum of both Alzheimer's patients and controls. The formation of these metabolites within distinct brain regions negatively correlated with the density of beta-amyloid deposits.

Decrease in pyrogenicity of muramyl dipeptide after coupling with luteinizing hormone-releasing hormone.

Muramyl dipeptide (MDP) and its adjuvant active derivative lysine-MDP (Lys-MDP) have been demonstrated to be pyrogenic and to induce endogenous pyrogen (EP) production in vivo and in vitro. It has recently been shown that immunologic castration can be achieved in mice by immunization with luteinizing hormone-releasing hormone (LHRH) directly conjugated by carbodiimide to Lys-MDP, termed LHRH-Lys-MDP (cdi), or with a linear monomeric MDP-linked molecule obtained by total synthesis, termed LHRH-Lys-MDP (s). These preparations were tested in the rabbit for their capacity to induce fever and were found to be devoid of pyrogenicity at dosage levels of Lys-MDP that induced fever. This decrease of pyrogenicity of Lys-MDP after coupling to LHRH seems to be related to the structure of the conjugate because the derivative LHRH-LysNH2-MDP exhibited the same pyrogenic activity as the free glycopeptide. Surprisingly, nonpyrogenic LHRH-Lys-MDP induced production of EP and interleukin-1 (IL-1) in vitro and increased in vivo modifications of metal levels attributed to the action of IL-1. Moreover, LHRH-Lys-MDP reduced the pyrogenic effect of an exogenous dose of EP.

Approaching criticality in polymer-polymer systems.

The interfacial width of polyolefins blends has been probed as a function of distance away from the critical point by using neutron reflectivity. For strongly immiscible polymer pairs, the width of the interface increases slowly when the degree of immiscibility is decreased and the interfacial width varies with the interaction parameter chi of the polymers. Closer to the critical point the dependence on the degree of miscibility becomes stronger and the way in which the interfacial width diverges, as criticality is approached, is related to both the chain length and chi. The self-consistent field theory numerical calculations, with the additional contribution due to capillary waves, provides a good description of the width of the interface between two polymer bulk phases in particular at intermediate values of the degree of immiscibility.

Analysis of the antigenic relationship of various derivatives of n-acetyl-muramyl-l-ala-d-isoglutamine (MDP), using anti-MDP antibodies.

Antibodies to N-acetyl-muramyl-L-alanyl-D-isoglutamine (MDP) were produced in rabbits by injection of MDP conjugated to various carriers [bovine gamma globulin (BGG), methylated BSA or sheep erythrocytes]. The anti-MDP was assayed by a direct enzyme-linked immunosorbent assay (ELISA) using horse radish peroxidase linked to MDP-Lys. Various derivatives of MDP were employed in an inhibition of ELISA for analysis of specificity of antibodies and study of the relationship of configuration to biological activity. The results confirmed previous findings that MDP alone is not immunogenic but can act as a hapten when conjugated to carriers. The antibodies were shown to be primarily directed against the muramyl residue. Modifications of this region of MDP yielded derivatives with weak reactivity against anti-MDP, while some changes of other regions had no effect on its antigenicity. Optical isomers of MDP had reduced activity as compared to MDP and polymeric MDP was a strong inhibitor. The structure and function relationship is discussed for some derivatives.

One-year follow-up of vaccine therapy in HIV-infected immune-deficient individuals: a new strategy.

Immunization of AIDS/ARC patients with autologous cells expressing HIV antigens, although providing clinical and biological benefits, fails to restore cellular immunity. The latter result is due partly to the antiproliferative effect of HIV-1 on activated T-cells (immune suppression), which leads to blockade of specific immune reactions. To overcome immune suppression, a new vaccine strategy was designed consisting of an immunization against HIV-1 combined with components of the T-cell-suppressive (antiproliferative) network. This new vaccine treatment proved to be innocuous in mice, monkeys, and two non-HIV-infected humans. A Phase I clinical trial was performed in six patients previously under cellular immunotherapy and still presenting a cellular immune defect. Preliminary results confirmed, after a 1-year follow-up of the patients, the safety of the new vaccine, which also partially restored the cellular immune response, including anti-HIV HLA-restricted cell-mediated cytotoxicity, delayed hypersensitivity to recall antigens, and proliferation of T-cells specifically activated by recall antigens.

Long-lasting rat growth enhancement by an immunoregulating synthetic peptide.

We have studied the long-lasting effects on rat weight gain of an active immunization with a peptide construction consisting of the covalent linkage of the 104-113 GH sequence to the 323-339 sequence of ovalbumin and to adjuvant muramyl dipeptide (MDP). The 104-113 GH sequence has already been identified as a potential epitope capable of enhancing growth hormome (GH) activity when complexed with the specific antibodies. Our results show that: a) 104-113 peptide antibodies after reacting with the endogenous GH produced a weight gain up to 7% higher than that observed in unimmunized rats; and b) the position of the adjuvant sequence in the molecule was critical for the immune response. Antibodies elicited in hypophysectomised rats had no effect on weight gain, thus confirming that the increase in hormonal activity is actually linked to the binding of specific peptide antibodies to endogenous GH.

HIV-1-induced immune suppression may result from autoimmune disorders including anti-SLWDQ autoantibodies.

We have previously unravelled the striking SLWDQ pentapeptide identity between HIV-1 env gp120 and the CD4 molecule. We show here that this pentapeptide is required for the functioning of the co-stimulatory MHC-CD4 signal in T4-cell activation since it suppresses antigen-induced T-cell proliferation. Moreover, concerning the MHC class II counterpart, the LNGQEETGVVSTN sequence which strongly inhibits T-cell immune activation is likely to be part of the functional site of the molecule. Interestingly the MHC/gp120 homology described by Young overlaps this MHC region. We further report that the gp120 SLWDQ peptide triggers an immune reaction which is both humoral (anti-SLWDQ antibodies) and cellular (CTLs against autologous targets carrying the pentapeptide) in HIV-1 infected individuals. Finally, anti-SLWDQ antibodies from patients sera purified by column chromatography strongly inhibit antigen-induced immune T-cell activation. This result led us to postulate that these antibodies found in high titers in HIV-1 infected individuals could contribute to set up the progressive systemic immune T-cell suppression characterizing AIDS.

Immunogenicity of combined anti-HIV and anti-suppressive vaccine preparations.

HIV-1 antigens generate in man both a humoral and cellular immune reaction. However, in ARC/AIDS patients, the cellular response is inhibited by HIV-1 which induces an antiproliferative (suppressive) effect on activated T cells. To overcome this inhibition and up-regulate the cellular response, we designed a new vaccine strategy directed both against HIV-1 and immunosuppression and we used an immunizing preparation composed of HIV-1 antigens combined with immunoregulatory peptides prepared in a biologically inactivated but immunogenic form. In mice, this preparation induced anti-HIV-1 antibodies and a cell-mediated cytotoxicity directed against H2 restricted cells carrying HIV-1 antigens.

Removal of gp160 induced bio-hazards for a safe AIDS vaccine candidate.

In the first AIDS vaccine trial, immunizing preparations were based on HIV-1 Env protein (gp160). Immunogenic properties of gp160 which trigger both a humoral and cellular immune response have since justified its use in various vaccine programs, both past and present. Many reports however have underlined deleterious effects on the immune system--anti-HIV-1 enhanced antibodies, anti-CD4 autoantibodies, and inhibition of T cell activation by HIV-1--particularly associated with the Env protein. The present study shows that gp160 presented in a biologically inactivated but immunogenic form, as used in our trial, could avoid these complications. Bio-hazards associated with gp160 which indeed could be removed by appropriate treatment of the native protein, should be taken into consideration in AIDS vaccine programs.

Percutaneous catheterization of the subclavian vein in hemophiliac patients: report of 47 cases.

Hemophiliac patients frequently require venous access, but usually their peripheral veins are collapsed and not puncturable. Several procedures employed to overcome this difficulty present a high complication rate. The authors report 47 cases of percutaneous subclavian vein catheterization (PSC) in hemophiliacs with an overall complication rate of 23%. Six patients (13%) had complications attributed to the coagulation disorder. The complications attributed to the coagulopathy and presenting major clinical importance occurred early in the series. After the improvement of the coagulation control with a routine AHG administration policy, the authors observed that among the last 22 patients there were 3 complications related to the coagulopathy, all of them without major clinical importance. The authors conclude that, in the lack of a safer access, PSC seems to be an acceptable procedure in hemophiliac patients, if employed with strict indication, accurate technique and rigorous control of the coagulopathhy.

[Immunisation of spontaneously hypertensive rats against angiotensin I]. / Immunisation du rat spontanément hypertendu contre l'angiotensine I.

Immunization against angiotensin I has been considered in comparison with the immunization against renin, in the spontaneously hypertensive rat (SHR). Among 6 different methods of immunization, two (AI-gluta-LPH and AI-Carbo-LPH coupling) permitted to obtain high levels of antibodies against angiotensin I (higher than 1/10,000), after four injections of 50 micrograms AI at three weeks interval. The titration of the antibodies was realized in radio-immuno-assay (RIA), with the determination of the cross-reactivity with AII by the same method. Characterization of the isotypes and the affinity calculation were realized with the ELISA method. The average level of antibodies is about 1/10,000 to 1/100,000, and the cross reactivity of the antibodies for AII is about 0.1 p. 100 in RIA. In ELISA, the study of the different isotypes shows a good maturation of the immune system with a sharp elevation of the IgG1 and IgG2 alpha isotypes, after 2 or 3 injections. The affinity of the antibodies purified by affinity chromatography is about 10.3 10(-9) M. The weekly measure of the arterial pressure during 6 months does not reveal at any moment a fall of pressure during the immunizations. The average pressure of the immunized group (209.4 +/- 23.8 mmHg, n = 40) is non significantly different from the average pressure of the mock group (208.5 +/- 22.6 mmHg, n = 10).

[Immunologic approach of the blockage of the renin angiotensin system in vivo]. / Approche immunologique du blocage du système rénine angiotensine in vivo.

To block the renin-angiotensin system by antibodies directed against renin or angiotensins is an old and recent goal. This goal can be attained by passive transfer of antibodies or by active immunization against the different molecules of the system. Only passive transfer of polyclonal antibodies directed against the native substrate (angiotensinogen) has been performed in rats. This acute blockade of angiotensinogen substrate availability decrease blood pressure about 30 mmHg in salt depleted rats. Passive transfer of anti-converting enzyme immunoglobulins has been already performed in rabbit and rat. It induced an immunoallergic reaction in the pulmonary capillary bed. Immunization against angiotensin II has been a powerful tool in the exploration of the role of the renin angiotensin system in hypertension. Passive and active immunization have been performed in different species: rabbit, rat. The majority of the results concerning the decrease in blood pressure was negative. However, some works reported positive results which could be related to the high affinity of antibodies for angiotensins. Passive and active immunizations against renin were also performed in different species: dog, pig, rat, rabbit, primates. The majority of the results concerning the decrease of blood pressure were positive, if species specificity of renin was taken into account. Recently passive transfer of polyclonal and monoclonal antibodies, directed against human renin have been performed in normotensive and hypertensive primates, demonstrating an acute fall in blood pressure comparable to that observed with converting enzyme inhibitors. Active immunization against human renin has also been performed in primates; and the chronic blockade of the renin-substrate reaction obtained in this way was associated with a significant decrease in blood pressure, aldosterone secretion and a disappearance of plasma renin activity. Unfortunately, such an active immunization was associated with an organ specific autoimmune disease within the kidney. In conclusion, passive and active immunization against the different proteins and peptides of the system offers specific models of blockade which can be compared with synthetic inhibitors of renin, converting enzyme and angiotensins. Therapeutic application of this immunological approach necessitates the verification of the total absence of autoimmune disease.

[Production and characterization of human antirenin antibodies prepared with synthetic immunogens]. / Production et caractérisation d'anticorps antirénine humaine préparés avec des immunogènes de synthèse.

Specific blockade of renin angiotensin system can be obtained both by enzymatic inhibitors and by passive and active immunization against renin. Recent studies have shown that synthetic peptides mimicking a protein segment can be used as immunogens to elicit antibodies which react with the parent molecule. In order to develop synthetic antirenin antigens we have selected peptidic sequences from human active renin and synthesized the corresponding peptides to produce antibodies able to recognize the entire human molecule and to inhibit its enzymatic activity. Three dimensional models of human renin were used to predict 17 putative epitopes. Then 18 peptides related to 13 potential antigenic determinants were synthesized by solid or liquid phase technic and 11 were shown to be antigenic when tested by their binding to several polyclonal and monoclonal human renin antibodies. The peptides were injected into rabbits and antisera tested by radio-immunoassay (RIA) and enzyme linked immunosorbent assay (ELISA) showed that nine peptides were immunogenic. Mainly, antiserum raised against the peptide mimicking the beta-hairpin 81-50 of active human renin which lies across the catalytic cleft, produced a 25 p. 100 inhibition of plasma renin activity at a 1: 50 dilution. Immunoglobulins, purified from antibodies raised against this epitopes, bound labelled renin and inhibited enzymatic activity of pure human renin on its synthetic tetradecapeptide substrate, in a dose dependent manner.

Immunological approach to blockade of the renin-substrate reaction.

To block the renin-substrate reaction by immunological tools is a long-standing dream. Since 1951 active immunization and the passive transfer of antirenin antisera have been successfully carried out in different species and in different experimental models of hypertension and normotension. These studies indicated that renin is a powerful immunogenic protein, capable of breaking down self-tolerance in different species. In this initial period the most significant results were obtained with hog renin. Passive transfer of antisera raised against hog renin or active immunization with hog renin was able to decrease blood pressure in renovascular or essential hypertension in dogs. Renin was semipurified and injected without adjuvant, however, since there was no method for determining plasma renin activity. Recently, complete purification of murine and human renin has allowed an extension of this approach, using the passive transfer of antirenin polyclonal antisera or monoclonal antibodies. Active immunization against pure human renin was successful in normotensive marmosets. This immunization with a nearly homologous renin in a primate model induced a significant decrease in blood pressure, associated with a complete disappearance of plasma renin activity. Unfortunately this powerful immunization was associated with an autoimmune disease that is specific for the kidney, related to self-recognition of the production site of renin by antibodies and lymphocytes. Similar results were reported with the use of mouse submandibular gland renin as an immunogen in spontaneously hypertensive rats (SHR). This manipulation decreased blood pressure in SHR to a level near that of normotensive Wistar-Kyoto control rats. However, again the animals showed a severe autoimmune disease of the kidney.(ABSTRACT TRUNCATED AT 250 WORDS)

Production and characterization of human renin antibodies prepared with synthetic immunogens.

Renin is an aspartyl protease that plays a key role in regulating blood pressure. The aim of this study was to use synthetic peptides to produce antibodies able to recognize the entire human renin molecule and to inhibit its enzymatic activity. Two three-dimensional renin models were used to predict 17 putative epitopes. Then 18 peptides related to 13 potential antigenic determinants were synthesized by solid- or liquid-phase technique, and 11 were shown to be antigenic when tested by their binding to several polyclonal human renin antibodies. The peptides were injected into rabbits, and antisera tested by radio-immunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) showed that nine peptides were immunogenic. The peptide related to the flap region which lies across the renin catalytic cleft showed potentially useful characteristics. Immunoglobulin G, purified from antibodies raised against this epitope, bound labelled renin and inhibited its enzymatic activity in a dose-dependent manner. This approach constitutes the basis for the development of a synthetic antirenin vaccine able to inhibit the renin-angiotensin system specifically.

Controlled sparse and percolating cross-linking in waterborne soft adhesives.

The effect of low levels of cross-linking on the adhesive and mechanical properties of waterborne pressure-sensitive adhesives was investigated. We have taken advantage of a core-shell latex particle morphology obtained by emulsion polymerization to create a heterogeneous structure of cross-links without major modification of the monomer composition. The latex particles comprise a shell containing cross-linkable diacetone acrylamide (DAAM) repeat units localized on the periphery of a slightly softer core copolymer of very similar composition. Adipic acid dihydrazide was added to the latex prior to film formation to react with DAAM repeat units and affect interfacial cross-linking between particles in the adhesive films. The honeycomb-like structure obtained after drying of the latex results in a good balance between the dissipative properties required for adhesion and the resistance to creep. The characterization of the mechanical properties of the films shows that the chosen cross-linking method creates a percolating lightly cross-linked network, swollen with a nearly un-cross-linked component. With this cross-linking method, the linear viscoelastic properties of the soft films are nearly unaffected by the cross-linking while the nonlinear tensile properties are greatly modified. As a result, the long-term shear resistance of the adhesive film improves very significantly while the peel force remains nearly the same. A simple rheological model is used to interpret qualitatively the changes in the material parameters induced by cross-linking.