Polymer-infiltrated ceramic CAD/CAM inlays and partial coverage restorations: 3-year results of a prospective clinical study over 5 years.

OBJECTIVE: The aim of this prospective clinical 5-year study was to evaluate the long-term behavior of monolithic computer-aided design and computer-aided manufacturing (CAD/CAM)-fabricated minimally invasive polymer-infiltrated ceramic network (PICN) inlays and partial coverage restorations (PCR). MATERIAL AND METHODS: Posterior teeth of 47 patients were restored with 103 restorations (45 inlays, 58 PCRs). After defect-oriented preparations, monolithic PICN restorations of VITA Enamic were fabricated with a CAD/CAM system (inEoS blue/CEREC inLab MCXL) and adhesively bonded (Variolink II). Clinical reevaluations were so far performed at baseline and 6, 12, 24, and 36 months after insertion according to modified United States Public Health Service (USPHS) criteria. Absolute failures were demonstrated by Kaplan-Meier survival rate and relative failures by Kaplan-Meier success rate. A logistic regression model was adjusted for modified USPHS criteria to investigate time and restoration effects (p < 0.05). RESULTS: After an observation time of 3 years, survival rates were 97.4% for inlays and 95.6% for PCRs. Three restorations had to be replaced due to clinically unacceptable fractures. Secondary caries and debonding were not observed. The 3-year Kaplan-Meier success rate was 84.8% for inlays and 82.4% for PCRs. The decrease in marginal adaption (p = 0.0005), increase in marginal discoloration (p < 0.0001), and surface roughness (p = 0.0005) over time were significant. Color match and anatomic form were excellent. No significant differences were found between both types of restorations for survival (p = 0.716) and success rate (p = 0.431). CONCLUSIONS: Minimally invasive PICN restorations showed a favorable clinical performance over an observation period of 36 months. However, clinical long-term data have to be awaited. CLINICAL RELEVANCE: PICN restorations are a suitable treatment option for posterior inlays and PCRs.

Management of shortened dental arches and periodontal health: 5-year results of a randomised trial.

In a multicentre randomised trial (German Research Association, grants DFG WA 831/2-1 to 2-6, WO 677/2-1.1 to 2-2.1.; controlled-trials.com ISRCTN97265367), patients with complete molar loss in one jaw received either a partial removable dental prosthesis (PRDP) with precision attachments or treatment according to the SDA concept aiming at pre-molar occlusion. The objective of this current analysis was to evaluate the influence of different treatments on periodontal health. Linear mixed regression models were fitted to quantify the differences between the treatment groups. The assessment at 5 years encompassed 59 patients (PRDP group) and 46 patients (SDA group). For the distal measuring sites of the posterior-most teeth of the study jaw, significant differences were found for the plaque index according to Silness and Löe, vertical clinical attachment loss (CAL-V), probing pocket depth (PPD) and bleeding on probing. These differences were small and showed a slightly more unfavourable course in the PRDP group. With CAL-V and PPD, significant differences were also found for the study jaw as a whole. For CAL-V, the estimated group differences over 5 years amounted to 0.27 mm (95% CI 0.05; 0.48; P = 0.016) for the study jaw and 0.25 mm (95% CI 0.05; 0.45; P = 0.014) for the distal sites of the posterior-most teeth. The respective values for PPD were 0.22 mm (95% CI 0.03; 0.41; P = 0.023) and 0.32 mm (95% CI 0.13; 0.5; P = 0.001). It can be concluded that even in a well-maintained.patient group statistically significant although minor detrimental effects of PRDPs on periodontal health are measurable.

Five-year results of a prospective randomised controlled clinical trial of posterior computer-aided design-computer-aided manufacturing ZrSiO4 -ceramic crowns.

The aim of this prospective randomised controlled clinical trial was to evaluate the clinical outcome of shrinkage-free ZrSiO4 -ceramic full-coverage crowns on premolars and molars in comparison with conventional gold crowns over a 5-year period. Two hundred and twenty-three patients were included and randomly divided into two treatment groups. One hundred and twenty-three patients were restored with 123 ZrSiO4 -ceramic crowns, and 100 patients received 100 gold crowns, which served as the control. All crowns were conventionally cemented with glass-ionomer cement. After an observation period of 6, 12, 24, 36, 48 and 60 months, the survival probability (Kaplan-Meier) for the shrinkage-free ZrSiO4 -ceramic crowns was 98·3%, 92·0%, 84·7%, 79% and 73·2% and for the gold crowns, 99%, 97·9%, 95·7%, 94·6% and 92·3%, respectively. The difference between the test and control group was statistically significant (P = 0·0027). The gold crowns showed a better marginal integrity with less marginal discoloration than the ceramic crowns. The most common failure in the ceramic crown group was fracture of the crown. The 60-month results of this prospective randomised controlled clinical trial suggest that the use of these shrinkage-free ZrSiO4 -ceramic crowns in posterior tooth restorations cannot be recommended.

Effect of veneering technique on the fracture resistance of zirconia fixed dental prostheses.

To compare the fracture resistance of zirconia 3-unit posterior fixed dental prostheses (FDPs) frameworks veneered with different veneering materials and techniques before and after artificial ageing. Forty-eight zirconia 3-unit FDPs, representing a missing first molar, were adhesively cemented on human teeth. The zirconia frameworks were randomly distributed according to the veneering materials and techniques into three groups, each containing 16 samples: group LV (layering technique/Vintage ZR), group LZ (layering technique/ZIROX) and group PP (CAD/CAM and press-over techniques/PressXZr). Half of each group was artificially aged through dynamic loading and thermocycling to simulate 5 years of clinical service. Afterwards, all specimens were tested for fracture resistance using compressive load. An analysis of variance (anova) was used to assess the effect of veneering ceramic and artificial ageing on fracture resistance (P < 0·05). Except for one minor cohesive chipping in group LV1, all specimens survived artificial ageing. The mean fracture resistance values (in Newton) of different non-aged (± s.d.)/aged (± s.d.) groups were as follows: LV0 2034 (± 401)/LV1 1625 (± 291); LZ0 2373 (± 718)/LZ1 1769 (± 136); and PP0 1959 (± 453)/PP1 1897 (± 329). Artificial ageing significantly reduced the fracture resistance in groups veneered with the layering technique (P < 0·05), whereas no significant effect was found in specimens veneered with the CAD/CAM and press-over techniques. All tested systems have the potential to withstand occlusal forces applied in the posterior region. The combination of the CAD/CAM and press-over techniques for the veneering process improved the overall stability after artificial ageing, relative to the layering technique.

Success rates of prosthetic restorations on endodontically treated teeth; a systematic review after 6 years.

The aims of this systematic review were to investigate the success rates of prosthetic restorations on endodontically treated teeth and their manner of failure. PubMed and hand literature searches were conducted on studies published until June 2012. Only clinical studies on human subjects referring to the success rates of prosthetic restorations on endodontically treated teeth with a follow-up period of at least 6 years were reviewed. A total of four studies were identified. Meta-analysis showed the success rate to be 92% (CI 82-98%) for single crowns on endodontically treated teeth and 79% (72-86%) for fixed dental prostheses. Only one study reported on removable dental prostheses with a success rate of 66%. Single crowns on teeth restored without posts demonstrated a success rate of 94% (CI 84-99%), whereas where posts were placed, the success rate was lower (92% CI 82-98%). Single crowns over cast post-and-cores and prefabricated posts showed success rates of 93% (CI 82-99%) and 94% (CI 90-97%), respectively; both differences were not statistically significant (significance level of 5%). The most common reason for failure was post-debonding. Single crowns seem to be the best treatment modality for endodontically treated teeth. However, due to the low number of studies included and their design, the results of this systematic review should be interpreted with caution. Further clinical studies are needed to provide high-quality evidence on the topic.

Prognosis of implants and fixed restorations after lateral sinus elevation: a literature review.

Aim of this review was to investigate the prognosis of implants inserted in augmented sinuses and fixed restorations supported by these implants. Special attention was given to the impact of grafting material, time of implant placement, residual bone height and type of fixed restoration. An electronic search in PubMed, the German database medpilot and the Cochrane Library was executed followed by supplementary manual search in relevant journals. The search was limited to human studies published up to November 2010. Only publications in English and German, in peer-reviewed journals, were considered. After the initial search and application of selection criteria on titles and abstracts, a full-text analysis of 67 articles was performed, out of which six prospective and three retrospective studies were finally included in the review. The heterogeneous properties of the identified articles did not allow systematic analysis of the data. Success rates of implants were between 96·3% and 100%, survival rates were between 75% and 100%, and survival rates of single crowns, splinted crowns and fixed partial dentures ranged between 96·4% and 100% after a follow-up of 12-101 months. Within the limits of this review, the prognosis of implants and fixed restorations seemed not to be influenced by the type of restorations, graft material, residual bone height and time of implant placement. However, conclusions of this review are based on studies with low level of evidence; therefore, careful interpretation is required. Multicentre randomised controlled clinical trials with sufficient statistical power concentrating on few factors are needed to reach sound conclusions.

Prognosis of immediately loaded implants and their restorations: a systematic literature review.

Immediate loading of oral implants has become popular because of the increasing demands of a shortened treatment time. This literature review evaluates the prognosis of immediately loaded implants and their restorations with immediate or delayed implant placement. Special attention was given to the impact of type of jaw, bone quality, implant length, time of implant placement and type of restoration. An electronic (PubMed) and a manual search in relevant journals were conducted until February 2012. Only publications in English, in peer-reviewed journals, were considered. Nine studies met the inclusion criteria: five studies dealt with fixed restorations, two studies with removable rehabilitation of edentulous jaws and two studies dealt with partially edentulous patients. Implant survival rates ranged from 95·8% to 100%, implant success rates in the treatment for the mandible from 79% to 100% and restoration survival rates for both jaws from 96·4% to 100%. Within the limits of this review, appropriate patient selection, primary implant stability, splinting of implants and the expertise of surgeons seem to be important for the prognosis of immediately loaded implants and their restorations. Good bone quality and use of long implants appear to play a role. However, careful interpretation is required because conclusions are based on articles with low level of evidence. While immediate loading of oral implants in the mandible shows encouraging and predictable results, further multicenter randomised controlled clinical trials with sufficient statistical power are needed to examine (i) the outcome of immediately loaded implants in the maxilla and (ii) the outcome of immediate loading of immediately placed implants.

The dental patient on oral anticoagulant medication: a literature review.

Specific diseases and medications may considerably influence the delivery of oral care and the course of dental therapy. The purpose of this literature review is to examine the relationship between oral anticoagulant medication and dental treatment. Electronic and manual searches were conducted for clinical studies in the English literature for the years 1988-2010. The review process provided a total of 110 pertinent literature references, out of which 38 studies dealt with oral anticoagulants and dental treatment. Different treatment strategies relative to dental periprocedural anticoagulation regimens have been identified, and their accompanying thromboembolic and bleeding risks are being presented and discussed. Regarding to what extent a safe and successful dental treatment in patients on anticoagulant medication is feasible, the level of evidence is lacking. Until high-level data are provided, an individualised treatment approach after consultation with the physician of the patient is highly recommended.

Prosthetic outcome of cement-retained implant-supported fixed dental restorations: a systematic review.

The aim of the article is to assess the current literature in terms of the prosthetic outcome of cement-retained implant-supported fixed restorations, as well as to determine the type of cement that can be recommended for clinical application. A review of the literature published up to May 2010 was conducted to identify clinical studies about cement-retained implant-supported fixed restorations. The search strategy applied was a combination of MeSH terms and free text words, including the following keywords: implants, implant-supported fixed dental prostheses (FDPs), bridges, implant-supported single crowns (SCs), cement-retained, cement fixation, cement, cementation, cement failure, retention, and loss of retention, technical complications, mechanical complications, prosthetic complication, retrievability and maintenance. Thirty-two studies met the inclusion criteria. The studies were divided into two categories: 15 short-term clinical studies with an observation period of less than 5 years, and 17 long-term clinical studies with an observation period of 5 years and more. The most common technical complications of cement-retained implant-supported fixed restorations were loss of retention, chipping and abutment screw loosening. The results of the current review revealed no guidelines about cement or cementation procedures. It may be stated that despite the questionable retrievability of cement-retained implant-supported fixed restorations, this treatment modality is a reliable and effective option, especially for implant-supported SCs and short-span FDPs. The literature does not provide accurate information about the clinical outcome of cement-retained implant-supported fixed restorations nor about the ideal type of cement that facilitates stability and maintains retrievability. Standardised randomised clinical trials will provide valuable information to this issue.

Music Community, Improvisation, and Social Technologies in COVID-Era Música Huasteca.

This article examines two interrelated aspects of Mexican regional music response to the coronavirus crisis in the música huasteca community: the growth of interactive huapango livestreams as a preexisting but newly significant space for informal community gathering and cultural participation at the onset of the coronavirus pandemic, and the composition of original verses by son huasteco performers addressing the pandemic. Both the livestreams and the newly created coronavirus disease (COVID) verses reflect critical improvisatory approaches to the pandemic in música huasteca. The interactive livestreams signaled an ad hoc community infrastructure facilitated by social media and an emerging community space fostered by Do-It-Yourself (DIY) activists. Improvised COVID-related verses presented resonant local and regional themes as a community response to a global crisis. Digital ethnography conducted since March 2020 revealed a regional burst of musical creativity coupled with DIY intentionality, a leveling of access to virtual community spaces, and enhanced digital intimacies established across a wide cultural diaspora in Mexico and the USA. These responses were musically, poetically, and organizationally improvisational, as was the overall outpouring of the son huasteco music inspired by the coronavirus outbreak. Son huasteco is a folk music tradition from the Huasteca, a geo-cultural region spanning the intersection of six states in central Mexico. This study examines a selection of musical responses by discussing improvisational examples in both Spanish and the indigenous language Nahuatl, and in the virtual musical communities of the Huasteca migrant diaspora in digital events such as "Encuentro Virtual de Tríos Huastecos," the "Huapangos Sin Fronteras" festival and competition, and in the nightly gatherings on social media platforms developed during the pandemic to sustain the Huastecan cultural expression. These phenomena have served as vibrant points of transnational connection and identity in a time where physical gatherings were untenable.

Results of a prospective randomized controlled trial of posterior ZrSiO(4)-ceramic crowns.

The aim of this randomized controlled clinical trial was to evaluate over a 5-year period, the clinical outcome of shrinkage-free ZrSiO(4)-ceramic (KaVo Everest HPC) full coverage crowns on posterior teeth in comparison with conventional gold crowns that served as the control. This study reports results of a 5-year study protocol up to 24 months. Patients totalling 224 were randomly divided into two treatment groups. Of these, 123 patients were restored with 123 Everest HPC crowns, fabricated by the Kavo Everest computer-aided manufacturing (CAM) procedure and 101 patients received 101 gold crowns, but two were excluded from analysis. All crowns were conventionally cemented with glass-ionomer cement. After an observation period of 6, 12 and 24 months, the prospective survival rates (Kaplan-Meier) for the KaVo Everest HPC crowns were 97.9%, 95.1% and 89.8% and for the gold crowns 100%, 94.8% and 92.7%, respectively. There were no significant differences between the two groups (P = 0.2). The 1-year failure rates were 4.9% for the KaVo Everest HPC crowns and 5.2% for the gold crowns. The 1-year cumulative risks for loss of vitality, secondary caries, fractures, loss of crown and extraction of abutment of the analyzed abutments (88) were 8.9%, 0%, 0%, 1.1% and 1.1%, respectively, for the gold crowns and 2.8%, 0%, 4.7%, 0% and 0.9%, respectively, for the ceramic crowns (107 analyzed abutments). No perfect marginal fit was shown by 49.5% of the evaluated ceramic crowns and 26.1% of the gold crowns. Only 1.9% of the KaVo Everest HPC crowns had a marginal crevice. In conclusion, Everest HPC crowns with an adequate occlusal tooth reduction of >1.5 mm are suitable for posterior restorations, but the marginal fit shows a potential for improvement.

Dental occlusion: a critical reflection on past, present and future concepts.

For nearly a century, the diversity of concepts about 'normal' and 'ideal' dental occlusal relationships has led to confusion in trying to describe the occlusion of any individual patient. In addition, a similar controversy arises when trying to formulate treatment plans for patients who need extensive dental restorations or orthodontic treatment. And finally, the application of occlusal concepts to patients with temporomandibular pain and dysfunction has created a third area of debate. Over the past few decades, however, an appreciable part of the tenacious dogmatic heritage of this topic has been challenged. As a result, the acceptance of morphological and functional variability of the stomatognathic system has gained increasing support, and this change has important consequences for modern dental practice. In this article, the past, present and future of the subject of occlusion will be considered.

Material characteristics of a novel shrinkage-free ZrSiO(4) ceramic for the fabrication of posterior crowns.

OBJECTIVE: To confirm the clinical applicability of a novel ZrSiO(4) (zircon) based shrinkage-free ceramic material, the flexural strength, fracture toughness and chemical solubility were tested. In addition, the fracture load of full crowns made from this material was tested after cyclic thermomechanical loading. METHODS: Flexural strength of 12 specimens was measured using a biaxial bending test. Fracture toughness was measured using 10 slotted box shaped specimens. The specimens were fractured using a universal testing machine; fracture loads were recorded. A chemical solubility test was performed in accordance with ISO norm 6872. Additionally, 32 ZrSiO(4) all-ceramic crowns were fabricated on extracted caries-free human molars. Sixteen Empress 2 and 16 PFM crowns served as a reference control. After artificial aging of 1.2 million cycles in the chewing simulator, the survival rate of the crowns was determined. The fracture load of all surviving specimens was obtained by loading the crowns until fracture in a universal testing machine. RESULTS: A flexural strength of 328MPa was found. The fracture toughness of the ZrSiO(4) ceramic was 5.16MPam(0.5). The chemical solubility amounted to 7.2microg/cm(2). All specimens survived the chewing simulation (survival rate: 100%); no crowns had to be re-cemented. A mean fracture strength of 1790N was found for Everest HPC for Empress 2 crowns, 1715N for Empress 2 crowns and 2416N for PFM crowns. Fracture loads of PFM crowns were significantly higher than for Empress 2 crowns (P=0.032) as well as ZrSiO(4)-crowns (P=0.007). There was no significant difference between ZrSiO(4)-crowns and Empress 2 crowns (P=0.743). SIGNIFICANCE: At the present stage, Everest HPC can be recommended for the fabrication of single crowns as an alternative to conventional PFM and other all-ceramic crowns, because its fracture strength exceeds average masticatory forces in the posterior region.

The GPI transamidase complex of Saccharomyces cerevisiae contains Gaa1p, Gpi8p, and Gpi16p.

Gpi8p and Gaa1p are essential components of the GPI transamidase that adds glycosylphosphatidylinositols (GPIs) to newly synthesized proteins. After solubilization in 1.5% digitonin and separation by blue native PAGE, Gpi8p is found in 430-650-kDa protein complexes. These complexes can be affinity purified and are shown to consist of Gaa1p, Gpi8p, and Gpi16p (YHR188c). Gpi16p is an essential N-glycosylated transmembrane glycoprotein. Its bulk resides on the lumenal side of the ER, and it has a single C-terminal transmembrane domain and a small C-terminal, cytosolic extension with an ER retrieval motif. Depletion of Gpi16p results in the accumulation of the complete GPI lipid CP2 and of unprocessed GPI precursor proteins. Gpi8p and Gpi16p are unstable if either of them is removed by depletion. Similarly, when Gpi8p is overexpressed, it largely remains outside the 430-650-kDa transamidase complex and is unstable. Overexpression of Gpi8p cannot compensate for the lack of Gpi16p. Homologues of Gpi16p are found in all eucaryotes. The transamidase complex is not associated with the Sec61p complex and oligosaccharyltransferase complex required for ER insertion and N-glycosylation of GPI proteins, respectively. When GPI precursor proteins or GPI lipids are depleted, the transamidase complex remains intact.

VISLISI trial, a prospective clinical study allowing identification of a new metalloprotease and putative virulence factor from Staphylococcus lugdunensis.

OBJECTIVE: Staphylococcus lugdunensis is a coagulase-negative staphylococcus that displays an unusually high virulence rate close to that of Staphylococcus aureus. It also shares phenotypic properties with S. aureus and several studies found putative virulence factors. The objective of the study was to describe the clinical manifestations of S. lugdunensis infections and investigate putative virulence factors. METHOD: We conducted a prospective study from November 2013 to March 2016 at the University Hospital of Strasbourg. Putative virulence factors were investigated by clumping factor detection, screening for proteolytic activity, and sequence analysis using tandem nano-liquid chromatography-mass spectrometry. RESULTS: In total, 347 positive samples for S. lugdunensis were collected, of which 129 (37.2%) were from confirmed cases of S. lugdunensis infection. Eighty-one of these 129 patients were included in the study. Bone and prosthetic joints (PJI) were the most frequent sites of infection (n=28; 34.6%) followed by skin and soft tissues (n=23; 28.4%). We identified and purified a novel protease secreted by 50 samples (61.7%), most frequently associated with samples from deep infections and PJI (pr 0.97 and pr 0.91, respectively). Protease peptide sequencing by nano-liquid chromatography-mass spectrometry revealed a novel protease bearing 62.42% identity with ShpI, a metalloprotease secreted by Staphylococcus hyicus. CONCLUSION: This study confirms the pathogenicity of S. lugdunensis, particularly in bone and PJI. We also identified a novel metalloprotease called lugdulysin that may contribute to virulence.

Marginal adaptation of different types of all-ceramic partial coverage restorations after exposure to an artificial mouth.

OBJECTIVES: To determine the influence of the preparation design and the dimensions of all-ceramic partial coverage restorations (PCR) on the marginal accuracy before and after masticatory simulation. METHODS: In this in vitro study 80 extracted human maxillary molars were restored with MOD inlay restorations and four different modified PCR restorations using a new press ceramic IPS e.max Press (IPS e.max Press VP 1989). The teeth were divided into five groups of 16 specimens each and prepared as follows: Group A received an MOD inlay preparation and Group B, C, D and E received modified PCR. The restorations were adhesively luted and exposed to a mastication simulator. The discrepancies of the marginal fit were examined on epoxy replicas before and after luting as well as after masticatory simulation at 200x magnification. RESULTS: The mean (geometrical) [95% confidence limits] marginal gap decreased from Group A to E before cementation (A-83[77-90]microm, B-68[65-70]microm, C-59[55-64]microm, D-56[52-61]microm, E-50[45-55]microm). Group A had significantly higher marginal gap values than group B (p = 0.017) and the other groups (p<0.0001). After cementation the marginal accuracy was recorded as following: A-103[93-114]microm, B-101[94-108]microm, C-93[89-98]microm, D-102[98-105]microm and E-99[96-102]microm. Cementation increased the marginal gap in groups B-E significantly (p<0.00001), not significantly in group A (p = 0.059). Artificial ageing (A-116[106-127]microm, B-114[109-120]microm, C-106[103-110]microm, D-109[100-118]microm and E-109[105-112]microm) led to further significant decrease of marginal accuracy in Group B (p = 0.029) and C (p = 0.026) only. After cementation and masticatory simulation of the ceramic restorations, the marginal gap values of Groups A, B, C, D and E did not significantly differ from each other (p = 1.00). CONCLUSIONS: The result of this in vitro study showed that IPS e.max Press can be used to fabricate all-ceramic inlays and PCR which meet the requirements in terms of a clinically acceptable marginal gap, irrespective of the preparation design used. However, the preparation design and dimensions of the restorations appeared to affect the initial marginal fit and flowing off of luting material during the cementation process. The factors responsible for these findings require further substantiation.

Antibacterial activity of secretolytin, a chromogranin B-derived peptide (614-626), is correlated with peptide structure.

Amongst the chromogranin B (CGB) derived fragments naturally generated in bovine chromaffin granules and detected in the extracellular space, we recently identified a major peptide corresponding to the 614-626 sequence of CGB. This peptide, named secretolytin, shared an interesting sequence homology with the lytic domain of cecropins and displayed a potent antibacterial activity. The aim of the present study was to determine the structural features of secretolytin necessary for this biological activity. Our results suggest that an alpha-helical amphipathic structure common to secretolytin, cecropins and pig myeloid antibacterial peptide may account for the antibacterial activity.

Study on agglutinating factors from flocculent Saccharomyces cerevisiae strains.

The lectin-like theory suggest that yeast flocculation is mediated by an aggregating lectinic factor. In this study we isolated an agglutinating factor, which corresponds to lectin, from whole cells by treating the flocculent wild-type Saccharomyces cerevisiae NCYC 625 strain and its weakly flocculent mutant [rho degrees ] with EDTA and two non-ionic surfactants (Hecameg and HTAC). The dialysed crude extracts obtained in this way agglutinated erythrocytes and this hemagglutination was specifically inhibited by mannose and mannose derivatives. However, SDS-PAGE profiles showed that the three reagents had different effects on the yeast cells. The non-ionic surfactants appeared to be the most efficient, as their extracts possessed the highest specific agglutinating activity. The products released by the wild-type strain presented a higher specific agglutinating activity than those released by the [rho degrees ] mutant. Purification of the agglutinating factor from extracts of both strains by affinity chromatography revealed two active bands of relative mass of 26 and 47 kDa on SDS-PAGE. Mass spectrometry analysis by MALDI-TOF, identified a 26 kDa band as the triose phosphate isomerase (TPI) whereas a 47 kDa band was identical to enolase. Edman degradation showed that the N-terminal sequences of these proteins were similar to TPI and enolase, respectively. The difference in the flocculation behaviour of the two strains is due to changes in the protein composition of the cell wall and in the protein structure involved in cell-cell recognition.

HistoCheck: rating of HLA class I and II mismatches by an internet-based software tool.

HLA polymorphism is a major barrier for hematopoietic stem cell and solid organ transplantation. To estimate the allogeneic potential between HLA-mismatched stem cell donor/recipient pairs, we recently proposed a matching score (dissimilarity index) that is based on the structural data of HLA class I molecules, and on the functional similarity of amino acids (AA). This first approach revealed new features about presumptive subtype allogenicities within the HLA-A*23 and A*24 groups. We have now developed an internet-based software tool ("HistoCheck") that is capable to assess the allogenicity (matching score) between any pair of clinically relevant HLA class I, and also class II, alleles. Newly described HLA sequences will be regularly integrated into the database according to the nomenclature for factors of the HLA system updates. The software is intended to be a first step for estimating the allogenicity of HLA mismatches in peculiar clinical settings, as long as there are no reliable in vitro or clinical studies available. The algorithm can later be modified according to functional data, for example, peptide-binding specificities. With the extension of the sequence similarity concept to all clinically relevant HLA class I and II loci, HistoCheck may contribute to prevent HLA mismatching being a matter of chance.

Antimicrobial chromogranins and proenkephalin-A-derived peptides: Antibacterial and antifungal activities of chromogranins and proenkephalin-A-derived peptides.

The secretory granules from adrenal medullary chromaffin cells contain a complex mixture of low-molecular mass constituents such as catecholamines, ascorbate, nucleotides, calcium, peptides, and several high-molecular mass water-soluble proteins including chromogranins and proenkephalin-A. These proteins are sequestered into secretory granules in which processing yields a large variety of peptides. These fragments are released into the extracellular space upon cell stimulation and are recovered in blood, lymph, cerebrospinal fluid, and synovial fluid. Some of them have biological activity on cells in an autocrine, paracrine, or endocrine fashion. In addition, we have shown that peptides with antimicrobial activity are present with the secretory chromaffin granules and demonstrated that they are released from stimulated chromaffin cells. We have shown that posttranslational modifications modulate the antimicrobial activities. For some peptides, using confocal laser microscopy, we have examined the interaction of the rhodaminated peptides with biological membranes. In addition, we have shown that chromofungin, the antifungal peptide corresponding to chromogranin A(47-66), can bind calmodulin in the presence of calcium and induce inhibition of calcineurin, a calmodulin-dependent enzyme. Because these antibacterial peptides are colocalized with catecholamines, they may be activated during stress, playing a role as a first protective barrier against bacterial infection, and thus act as factors of the innate immunity shortly after infection and before the induction and mobilization of an adaptative immune system.