A fibre and phenolic-rich flour from Isabel grape by-products with stimulatory effects on distinct probiotics and beneficial impacts on human colonic microbiota in vitro.

This study evaluated the effects of a fibre and phenolic-rich flour (IGF) prepared from Isabel grape by-products on the growth and metabolism of different probiotics and distinct bacterial populations part of the human intestinal microbiota during an in vitro colonic fermentation. IGF was submitted to simulated gastrointestinal digestion before use in the experiments. IGF favoured the growth of the probiotics Lactobacillus acidophilus La-05, L. casei L-26 and Bifidobacterium lactis Bb-12, with viable counts of >7 log CFU per ml, as well as caused decreases in pH values and increases in organic acid production in the growth medium during 48 h of cultivation. IGF increased the population of beneficial micro-organisms forming the human intestinal microbiota, particularly Lactobacillus spp., decreased the pH values, and increased the lactic acid and short-chain fatty acid (acetic, butyric and propionic acids) production during 24 h of in vitro colonic fermentation. These results indicate the potential prebiotic effects of IGF, which should represent a novel sustainable added-value ingredient with functional properties and gut-health benefits.

Thyroid hormone responsive QTL and the evolution of paedomorphic salamanders.

The transformation of ancestral phenotypes into novel traits is poorly understood for many examples of evolutionary novelty. Ancestrally, salamanders have a biphasic life cycle with an aquatic larval stage, a brief and pronounced metamorphosis, followed by a terrestrial adult stage. Repeatedly during evolution, metamorphic timing has been delayed to exploit growth-permissive environments, resulting in paedomorphic salamanders that retain larval traits as adults. We used thyroid hormone (TH) to rescue metamorphic phenotypes in paedomorphic salamanders and then identified quantitative trait loci (QTL) for life history traits that are associated with amphibian life cycle evolution: metamorphic timing and adult body size. We demonstrate that paedomorphic tiger salamanders (Ambystoma tigrinum complex) carry alleles at three moderate effect QTL (met1-3) that vary in responsiveness to TH and additively affect metamorphic timing. Salamanders that delay metamorphosis attain significantly larger body sizes as adults and met2 explains a significant portion of this variation. Thus, substitution of alleles at TH-responsive loci suggests an adaptive pleiotropic basis for two key life-history traits in amphibians: body size and metamorphic timing. Our study demonstrates a likely pathway for the evolution of novel paedomorphic species from metamorphic ancestors via selection of TH-response alleles that delay metamorphic timing and increase adult body size.

Excited state characterization of a polymeric indigo.

A comprehensive investigation of the solution photophysics of a 5,5'-methylene-bridged polymeric indigo, a statistical copolymer consisting of indigo and N-acetylindigo units, was performed in organic solvents at room temperature and further compared with indigo. A complete spectral and photophysical characterization based on photoacoustic calorimetry, steady-state and time-resolved fluorescence data was undertaken. A fluorescence quantum yield of 0.00037 and an intersystem crossing singlet-to-triplet quantum yield of 0.006 (close to the value for indigo) were obtained, leading to a value of 0.9936 for the S(1) → S(0) internal conversion (IC) quantum yield. Spectral and photophysical characteristics similar to indigo were obtained with, however, a special signature: it (mainly) decays single exponentially (in contrast with indigo, found to decay bi-exponentially), with a decay time value of 40-50 ps and an even more efficient S(1) → S(0) IC deactivation channel, related to an efficient energy migration within an energetic ladder of the polymer chromophoric segments. The photochemistry of this polymer, namely the degradation under light excitation, was also investigated and the obtained photoreaction quantum yield (ϕ(R)) in DMF was found to be 0.003, which is lower than the previously determined value for indigo in the same solvent (ϕ(R) = 0.0078). The overall data indicate that although the polymer and indigo have a close finger-print, the former is more stable which is suggested to be due to the additional intramolecular energy transfer process (within different chromophoric units) found with the polymer.

Evaluation of the safety and immunogenicity of Plasmodium falciparum apical membrane antigen 1, merozoite surface protein 1 or RTS,S vaccines with adjuvant system AS02A administered alone or concurrently in rhesus monkeys.

In an effort to broaden the immune response induced by the RTS,S/AS02(A),vaccine, we have evaluated the immunogenicity of the RTS,S antigen when combined with MSP1(42) and with AMA1, antigens derived from the asexual blood stage. The objectives of this study were (i) to determine whether MSP1(42) and AMA1 vaccines formulated with the AS02(A) Adjuvant System were safe and immunogenic in the rhesus monkey model; (ii) to investigate whether MSP1(42) or AMA1 induced immune interference to each other, or to RTS,S, when added singly or in combinations at a single injection site; (iii) in the event of immune interference, to determine if this could be reduced when antigens were administered at separate sites. We found that MSP1(42) and AMA1 were safe and immunogenic, eliciting antibodies, and Th1 and Th2 responses using IFN-gamma and IL-5 as markers. When malaria antigens were delivered together in one formulation, MSP1(42) and RTS,S reduced AMA1-specific antibody responses as measured by ELISA however, only MSP1(42) lowered parasite growth inhibitory activity of anti-AMA1 antibodies as measured by in vitro growth inhibition assay. Unlike RTS,S, MSP1(42) significantly reduced AMA1 IFN-gamma and IL-5 responses. MSP1(42) suppression of AMA1 IFN-gamma responses was not seen in animals receiving RTS,S+AMA1+MSP1(42) suggesting that RTS,S restored IFN-gamma responses. Conversely, AMA1 had no effect on MSP1(42) antibody and IFN-gamma and IL-5 responses. Neither AMA1 alone or combined with MSP1(42) affected RTS,S antibody or IFN-gamma and IL-5 responses. Immune interference by MSP1(42) on AMA1 antibody responses was also evident when AMA1, MSP1(42) and RTS,S were administered concurrently at separate sites. These results suggest that immune interference may be complex and should be considered for the design of multi-antigen, multi-stage vaccines against malaria.

Normal hospital and low-bacterial diet in patients with cytopenia after intensive chemotherapy for hematological malignancy: a study of safety.

BACKGROUND: The purpose of this randomized, controlled pilot study is to address the question whether normal hospital diet (NHD) is safe when compared with low-bacterial diet (LBD) given to prevent infections in cytopenic patients who receive antimicrobial prophylaxis (AP). PATIENTS AND METHODS: The patients were randomized into two groups: one group to receive AP and LBD, the other to receive the same AP and NHD. The primary outcome parameter is colonization of the digestive tract with aerobic gram-negative bacilli and yeasts. Secondary outcome parameters were infections and total societal costs. RESULTS: No statistically significant differences between treatment groups were observed regarding the primary outcome parameter, gut colonization by yeasts or gram-negative bacilli, or infections, use of antimicrobials, days with fever and total societal costs. CONCLUSION: On the basis of the results of this pilot study, NHD appears to be as safe as LBD in patients with chemotherapy-induced cytopenia. Furthermore, the results indicate that LBD may offer no additional benefit as an infection preventive measure to the measures already implemented, such as AP. Thus, a larger randomized study, powered adequately to determine noninferiority of NHD to LBD is warranted and safe to be carried out.

Photophysics of an indigo derivative (keto and leuco structures) with singular properties.

Spectral and photophysical properties of the indigo derivative Cibalackrot in keto and reduced (leuco) forms were studied by absorption spectra, fluorescence and pulse radiolysis and compared with the structurally similar indigo. With the keto form of this dye, fluorescence (phiF = 0.76) and intersystem crossing (phiT = 0.11) are dominant, whereas with indigo, efficient internal conversion (phiIC = 0.99) is observed, probably involving proton transfer through intramolecular hydrogen bonds. With Cibalackrot, this pathway is blocked, supporting the above model for indigo. With the reduced form of Cibalackrot, more than 98% of the absorbed quanta are dissipated through S1 approximately --> S0 internal conversion, which contrasts with leuco-indigo, where fluorescence (phiF = 0.35), internal conversion (phiIC = 0.53) and intersystem crossing (phiT = 0.125) are found to be competitive. In addition, a synthetic precursor of Cibalackrot (preCiba) was also investigated. This has a rigid molecular structure (with a moiety identical to Cibalackrot and the other to indigo), but intra- or intermolecular proton transfer is allowed between adjacent carbonyl and N-H groups. With this precursor in its keto structure the photophysical parameters are generally very close to those of the keto form of indigo, and different from those of Cibalackrot. A more detailed investigation of the time-decay profiles of preCiba in dioxane (and with added water and D2O) has shown that these follow biexponential laws with a shorter component of 14-25 ps, which appears associated with a risetime at longer wavelength emissions (and to a positive preexponential at shorter emission wavelengths) and a longer lived (decay) component of 104-130 ps. In the steady-state spectra of preCiba, the variation with temperature reveals a blue shift of the emission maxima, which is interpreted as the presence (simultaneous emission) of two species (keto and enol) in the excited state. Indigo and deuterated indigo are also found to present a similar behavior. The overall data are interpreted as to be due to an excited-state process involving the proton transfer between keto and enol forms. Rate constants with values of 7 x 10(10) s-1 for preCiba and 1.6 x 10(11) s-1 for deuterated indigo were obtained. This inverse isotope effect is justified on the basis of the proposed model for proton-transfer excited-state deactivation.

Characterization of antibody responses elicited by human immunodeficiency virus type 1 primary isolate trimeric and monomeric envelope glycoproteins in selected adjuvants.

We previously reported that soluble, stable YU2 gp140 trimeric human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein immunogens could elicit improved breadth of neutralization against HIV-1 isolates compared to monomeric YU2 gp120 proteins. In this guinea pig immunization study, we sought to extend these data and determine if adjuvant could quantitatively or qualitatively alter the neutralizing response elicited by trimeric or monomeric immunogens. Consistent with our earlier studies, the YU2 gp140 immunogens elicited higher-titer neutralizing antibodies against homologous and heterologous isolates than those elicited by monomeric YU2 gp120. Additionally, the GlaxoSmithKline family of adjuvants AS01B, AS02A, and AS03 induced higher levels of neutralizing antibodies compared to emulsification of the same immunogens in Ribi adjuvant. Further analysis of vaccine sera indicated that homologous virus neutralization was not mediated by antibodies to the V3 loop, although V3 loop-directed neutralization could be detected for some heterologous isolates. In most gp120-inoculated animals, the homologous YU2 neutralization activity was inhibited by a peptide derived from the YU2 V1 loop, whereas the neutralizing activity elicited by YU2 gp140 trimers was much less sensitive to V1 peptide inhibition. Consistent with a less V1-focused antibody response, sera from the gp140-immunized animals more efficiently neutralized heterologous HIV-1 isolates, as determined by two distinct neutralization formats. Thus, there appear to be qualitative differences in the neutralizing antibody response elicited by YU2 gp140 compared to YU2 monomeric gp120. Further mapping analysis of more conserved regions of gp120/gp41 may be required to determine the neutralizing specificity elicited by the trimeric immunogens.

Cost-effectiveness of Chlamydia antibody tests in subfertile women.

BACKGROUND: For the evaluation of tubal function, Chlamydia antibody testing (CAT) has been introduced as a screening test. We compared six CAT screening strategies (five CAT tests and one combination of tests), with respect to their cost-effectiveness, by using IVF pregnancy rate as outcome measure. METHODS: A decision analytic model was developed based on a source population of 1715 subfertile women. The model incorporates hysterosalpingography (HSG), laparoscopy and IVF. To calculate IVF pregnancy rates, costs, effects, cost-effectiveness and incremental costs per effect of the six different CAT screening strategies were determined. RESULTS: pELISA Medac turned out to be the most cost-effective CAT screening strategy (15 075 per IVF pregnancy), followed by MIF Anilabsystems (15 108). A combination of tests (pELISA Medac and MIF Anilabsystems; 15 127) did not improve the cost-effectiveness of the single strategies. Sensitivity analyses showed that the results are robust for changes in the baseline values of the model parameters. CONCLUSIONS: Only small differences were found between the screening strategies regarding the cost-effectiveness, although pELISA Medac was the most cost-effective strategy. Before introducing a particular CAT test into clinical practice, one should consider the effects and consequences of the entire screening strategy, instead of only the diagnostic accuracy of the test used.

The virtual articulator.

The virtual articulator (VA) is intended for use as a tool for the analysis fo the complex static and dynamic occlusal relations. Its chief application is in individual diagnosis and the simulation of the mechanical articulator. As entry data, the VA requires digital, 3-dimensional representations of the jaws and patient-specific data on jaw movement. It then simulates jaw movement and provides a dynamic visualization of the occlusal contacts. If no patient-specific data are available, the modus operandi of the mechanical articulator can be simulated. Virtual reality (VR) techniques applied in computer-aided diagnosis are revolutionizing medical routine in many disciplines. Particularly in the field of dental engineering, VR combined with CAD/CAM technology has great potential in implant planning and design in terms of greater precision and shorter treatment duration. The geometry of teeth can be digitized, the implant models can be designed with specialized CAD systems, and the final implants can be manufactured with a CNC machine. In this context, the virtual articulator completes the digital manufacturing process.

Efficacy of RTS,S/AS02 malaria vaccine against Plasmodium falciparum infection in semi-immune adult men in The Gambia: a randomised trial.

BACKGROUND: RTS,S/AS02 is a pre-erythrocytic malaria vaccine based on the circumsporozoite surface protein of Plasmodium falciparum fused to HBsAg, incorporating a new adjuvant (AS02). We did a randomised trial of the efficacy of RTS,S/AS02 against natural P. falciparum infection in semi-immune adult men in The Gambia. METHODS: 306 men aged 18-45 years were randomly assigned three doses of either RTS,S/AS02 or rabies vaccine (control). Volunteers were given sulfadoxine/pyrimethamine 2 weeks before dose 3, and kept under surveillance throughout the malaria transmission season. Blood smears were collected once a week and whenever a volunteer developed symptoms compatible with malaria. The primary endpoint was time to first infection with P. falciparum. Analysis was per protocol. FINDINGS: 250 men (131 in the RTS,S/AS02 group and 119 in the control group) received three doses of vaccine and were followed up for 15 weeks. RTS,S/AS02 was safe and well tolerated. P. falciparum infections occurred significantly earlier in the control group than the RTS,S/AS02 group (Wilcoxon's test p=0.018). Vaccine efficacy, adjusted for confounders, was 34% (95% CI 8.0-53, p=0.014). Protection seemed to wane: estimated efficacy during the first 9 weeks of follow-up was 71% (46-85), but decreased to 0% (-52 to 34) in the last 6 weeks. Vaccination induced strong antibody responses to circumsporozoite protein and strong T-cell responses. Protection was not limited to the NF54 parasite genotype from which the vaccine was derived. 158 men received a fourth dose the next year and were followed up for 9 weeks; during this time, vaccine efficacy was 47% (4-71, p=0.037). INTERPRETATION: RTS,S/AS02 is safe, immunogenic, and is the first pre-erythrocytic vaccine to show significant protection against natural P. falciparum infection.

Absorption and disposition of levocetirizine, the eutomer of cetirizine, administered alone or as cetirizine to healthy volunteers.

The primary objective of the present study was to compare the absorption and disposition of levocetirizine, the eutomer of cetirizine, when administered alone (10 mg) or in presence of the distomer. An additional objective was also to investigate the configurational stability of levocetirizine in vivo in humans. The study was performed in a randomized, two-way cross-over, single-dose design with a wash-out phase of 7 days between the two periods. A total of 12 healthy male and 12 healthy female volunteers were included in the study. Bioequivalence can be concluded from the analysis of the pharmacokinetic parameters of levocetirizine when administered alone or as the racemate cetirizine. No chiral inversion occurs in humans when levocetirizine is administered, i.e. there is no formation of the distomer. When comparing the pharmacokinetic characteristics of levocetirizine and the distomer, the apparent volume of distribution of the eutomer is significantly smaller than that of the distomer (0.41 and 0.60 L/kg, respectively). For an H1-antagonist a small distribution volume can be considered as a positive aspect, both in terms of efficacy and safety. Moreover the non-renal clearance of levocetirizine is also significantly lower than that of the distomer (9.70 and 28.70 mL/min, respectively), which constitutes an additional positive aspect particularly as far as metabolism-based drug interactions are concerned. The information collected in the present study on the pharmacokinetics of levocetirizine and the distomer provide additional reasons for eliminating the distomer and developing levocetirizine as an improvement on cetirizine.

Characterization of HIV-specific proliferative T cell responses in HIV-infected persons.

Virus-specific helper T cell responses are thought to be an important host defense in HIV infection. The proliferative responses to HIV p24, p55, and gp120 were tested in a cohort of 27 HIV-infected subjects. Vigorous proliferative responses directed at the Gag protein with stimulation indices in excess of 6 were detected in 10 of the individuals tested but an Env-specific response was present in only 1 subject. Viral load and proliferative activity to Gag were inversely correlated in untreated individuals. Proliferation was also observed in some individuals treated in the chronic phase of infection, and responses were maintained over time in the absence of detectable viremia. Positive proliferative responses could also occasionally be detected in treated persons with CD4(+) cell counts below 200/microl. Thus, vigorous Gag-specific proliferative responses are present in a minority of HIV-infected individuals and can be detected in individuals receiving highly active antiretroviral therapy at advanced disease stages. Proliferative responses are maintained for an extended time period in the presence of antiviral therapy.

LAHYSTOTRAIN development and evaluation of a complex training system for hysteroscopy.

Hysteroscopy has already become an irreplaceable method in gynaecoloic diagnosis and therapy. In the diagnostic case the hysteroscope with a 30 degrees optic is insert transvaginally, in the therapeutic case the resectoscope with a 12 degrees optic is used. The endoscopic intervention requires special surgical skills for endoscope handling and remote instrument control. To acquire these skills currently hands-on training in clinical praxis has become standard, which is linked with higher danger for the women. To overcome current drawbacks of traditional training methods the European project LAHYSTOTRAIN was set up, that tries to combine Virtual Reality (VR), Multimedia (MM) technology, and Intelligent Tutoring Systems (ITS) to develop an alternative training system for hysteroscopic interventions. The first prototype of the LAHYSTOTRAIN demonstrator has been shown on several European conferences. An evaluation of the system was performed, with the idea, to collect feedback and impressions, that should be considered in further developments. This paper presents the LAHYSTOTRAIN prototype and the results of these evaluations.

ICAPS an integrative computer-assisted planning system for pedicle screw insertion.

Robot Assisted Surgery (RAS) Systems win more and more recognition in the field of orthopaedics. Especially in Hip Surgery RAS has proved to be suited for application in medical routine. Often Robot Assisted Surgery Systems consist of a planning- and an interoperative component. According to specifications done with the planning software the tools are driven. Benefits of the robot assisted surgery should be higher precision and a better surgical outcome. In the co-operation project of several Fraunhofer Institutes "RoMed" (Robots and Manipulators for Medical Application) an exemplary application of robot aided spine surgery is developed [1]. The planning software used in this context is proposed in this article.

Efficacy of recombinant circumsporozoite protein vaccine regimens against experimental Plasmodium falciparum malaria.

After initial successful evaluation of the circumsporozoite-based vaccine RTS,S/SBAS2, developed by SmithKline Beecham Biologicals with the Walter Reed Army Institute of Research, protective efficacy of several regimens against Plasmodium falciparum challenge was determined. A controlled phase 1/2a study evaluated 1 or 2 standard doses of RTS,S/SBAS2 in 2 groups whose members received open-label therapy and 3 immunizations in blinded groups who received standard, one-half, or one-fifth doses. RTS,S/SBAS2 was safe and immunogenic in all groups. Of the 41 vaccinees and 23 control subjects who underwent sporozoite challenge, malaria developed in 7 of 10 who received 1 dose, in 7 of 14 who received 2 doses, in 3 of 6 who received 3 standard doses, in 3 of 7 who received 3 one-half doses, in 3 of 4 who received 3 one-fifth doses, and in 22 of 23 control subjects. Overall protective efficacy of RTS,S/SBAS2 was 41% (95% confidence interval, 22%-56%; P=.0006). This and previous studies have shown that 2 or 3 doses of RTS,S/SBAS2 protect against challenge with P. falciparum sporozoites.

Planning system for computer assisted total knee replacement.

Total knee replacement (TKR) is a common orthopaedic surgical intervention and includes the removal of bone sections from the end of the femur and the top of the tibia for replacement by prosthetic components. Pain relief and functional improvement are predictable clinical results. But the accuracy of the alignment affects the surgical outcome and the longevity of the prosthesis. Hence, current total knee implantation systems attempt to align the knee joint in the mechanical axis for placement of the total knee components. These approaches use templates and plain radiographs for preoperative planning and alignment devices for bone cuts. To overcome the inherent inaccuracy of the presently used systems a computer-assisted planning system has been developed delivering the necessary control data for the intraoperative surgical robot system.

LAHYSTOTRAIN intelligent training system for laparoscopy and hysteroscopy.

Rapid developments in the medical field, as an expanding knowledge base and emerging new technologies require continuing medical education to achieve life long learning and to keep the surgeons up to date. Consequently, specific training is necessary to guarantee qualification of the surgeons. The goal of LAHYSTOTRAIN is to overcome the current drawbacks of traditional training methods for laparoscopic/hysteroscopic procedures. A computer-assisted simulator for training and quality control in laparoscopy and hysteroscopy is developed using Virtual Reality (VR), Multimedia (MM) technology, and Intelligent Tutoring Systems (ITS).

Rhinosurgical therapy planning via endonasal airflow simulation.

Nowadays, Computational Fluid Dynamics (CFD) methods play an important part in the production process of the automotive industry. Progress in recent years has made possible highly sophisticated airflow-simulation models that are used in engineering for optimization and verification of aerodynamics. The key purpose of the Simulation Tool for Airflow in the human Nose (STAN), developed at the Darmstadt University of Technology in cooperation with the University Hospital in Greifswald, is to use these techniques to support the rhinosurgeon in diagnosis and planning of therapy (Frühauf T, Mlynski G. Simulation and visualization of the air flow in the human nose. Proceedings of the First World Congress on Computational Medicine, Austin, Texas, 1994). A system has been developed that realizes a three-dimensional (3D) reconstruction of the endonasal cavities based on computer tomography (CT) scans. This semiautomatic reconstruction method requires minimal manual intervention. The surface model is used to create an unstructured 3D volume mesh suitable for finite volume simulations. In this way, an individual simulation based on patient-specific data can be realized. At the University Hospital in Greifswald, experimental investigations and measurements are made in nasal models to verify the simulation result. The goal of this project is to investigate individual nasal complaints and to detect respiratory disorders. The surgeon should be able to simulate the disordered respiration before performing a surgical procedure, and thereby increase the effectiveness of surgical planning.

Simian immunodeficiency virus evades a dominant epitope-specific cytotoxic T lymphocyte response through a mutation resulting in the accelerated dissociation of viral peptide and MHC class I.

The ability of an AIDS virus to escape from immune containment by selective mutation away from recognition by CTL was explored in simian immunodeficiency virus of macaques (SIVmac)-infected rhesus monkeys. CTL recognition of a previously defined common viral mutation in an immunodominant SIVmac Gag epitope was evaluated. CTL were assessed for their ability to recognize a SIVmac Gag protein with a single residue 2 (T --> A) replacement in the minimal epitope peptide bound by the MHC class I molecule Mamu-A*01. SIVmac Gag-specific CTL lysed Mamu-A*01+ target cells infected with recombinant vaccinia virus expressing the wild-type but not the mutant Gag protein. In addition, CTL recognized the mutant epitope peptide less efficiently than the wild-type virus peptide. In studies to determine the mechanism by which the mutant virus evaded CTL recognition, this peptide was shown to bind Mamu-A*01 in a manner that was indistinguishable from the wild-type peptide. However, experiments in which an increasing duration of delay was introduced between peptide sensitization of target cells and the assessment of these cells as targets in killing assays suggest that the mutant peptide with a T --> A replacement had a higher off-rate from Mamu-A*01 than the wild-type peptide did. Therefore, these findings suggest that AIDS viruses can evade virus-specific CTL responses through the accelerated dissociation of mutant peptide from MHC class I.