Bacteroides fragilis: A whole MALDI-based workflow from identification to confirmation of carbapenemase production for routine laboratories.

Bacteroides fragilis is a frequent anaerobic pathogen and can cause severe infections. Resistance to carbapenems, associated with the cfiA gene encoded carbapenemase, represents an emerging problem. To date, no rapid methods are available to detect and confirm this resistance mechanism in routine laboratories, and the missed recognition of carbapenemase-producing strains can lead to therapeutic failures. In this study we have investigated a whole MALDI-TOF MS-based workflow to detect carbapenemase-producing B. fragilis, using the largest set of B. fragilis clinical isolates ever tested. The presence of the cfiA gene was predicted by MALDI subtyping into Division I (cfiA-negative) or Division II (cfiA-positive). The carbapenemase activity in cfiA-positive strains was confirmed by a MALDI-TOF MS imipenem hydrolysis assay (MBT STAR-Carba, Bruker Daltonik, Germany), that was further used for a characterization of the strains in terms of cfiA expression level. The validity of MALDI subtyping was verified by PCR for the cfiA gene, while results of MALDI hydrolysis assay were compared to conventional methods for susceptibility testing and carbapenemase detection (Carba-NP and disk diffusion synergy test). A genetic analysis of the IS elements upstream cfiA was performed, for the evaluations regarding the expression level of cfiA. A total of 5300 B. fragilis isolates (406 from Bologna, Italy, and 4894 from Dortmund, Germany) were identified and subtyped by MALDI-TOF MS, yielding 41/406 (10.1%) strains from Bologna and 374/4894 (7.6%) from Dortmund to belong to Division II. Molecular verification by PCR for the cfiA gene on a subset of strains confirmed the MALDI typing results in all cases (sensitivity and specificity of 100%). MBT STAR-Carba assay detected the carbapenemase activity in all of the 70 cfiA-carrying strains tested. Moreover, it allowed distinct separation into slow (59) and fast (11) imipenem hydrolyzers corresponding to cfiA expression levels as well as to low or high MICs for carbapenems, respectively. Among the 11 cfiA-positive strains with high carbapenem MIC, only 7 harboured IS elements upstream the carbapenemase gene showing low expression level as well. The MALDI-TOF MS-based workflow was superior to the currently available phenotypic methods for carbapenemase detection as it proved to be more sensitive and accurate than Carba NP and disk diffusion synergy test. The whole MALDI-TOF MS-based workflow allows an accurate identification of B. fragilis clinical strains with reliable classification into Division I/II, and confirmation of the carbapenemase-production, together with estimation of carbapenemase activity, within less than 2 h. This may be of particular interest for early therapeutical decisions in life-threatening infections.

Effectiveness of bionator therapy for Class II malocclusions: a comparative long-term study.

OBJECTIVE: The goal of this retrospective study was to examine the effectiveness of isolated bionator therapy in Class II patients both longitudinally and over the long term. We aimed to determine whether skeletal and/or dentoalveolar reactions differ in relationship to the Angle Class (II, Division 1 vs. II, Division 2). PATIENTS AND METHODS: A total of 50 juvenile patients with Class II malocclusion (♀ n = 26, ♂ n = 24) were included. A total of 24 patients presented an Angle Class II, Division 1 and 26 an Angle Class II, Division 2. We compared the patients' lateral cephalograms taken at the beginning of treatment (t1: 10.1 years), after active therapy (t2: 13.8 years), and at the end of retention (t3: 16.4 years) analyzing the following cephalometric parameters: SNA, SNB, ANB, ANB(ind), SNPog, Wits appraisal, U1-SN, U1-SpP, L1-MeGo, interincisal angle. Mean and standard deviations of each of the variables were calculated. Differences between t1-t2 and t2-t3 were tested for statistical significance. Changes in the variables were then analyzed biometrically for specific differences in terms of Angle Class (II, Division 1 vs. II, Division 2). RESULTS: Between t1 and t2, SNB (p = 0.000) and SNPog (p = 0.000) increased significantly, as did ANB (p = 0.000), while the difference between ANB and ANB(ind) (p = 0.000) and Wits appraisal (p = 0.000) decreased significantly. The dentoalveolar variables U1-SN, U1-SpP, and the interincisal angle changed significantly in both groups. The inclination of the upper incisors was corrected by retrusion in the Class II, Division 1 and by protrusion in the Class II, Division 2 group. Only marginal changes in all variables between t2 and t3 were observed. CONCLUSION: A significant skeletal effect (even in long-time stability) through bionator treatment could be confirmed in this study of Class II, Divisions 1 and 2 patients. The desired effect on the upper front teeth was realized, and there was no appreciable dentoalveolar compensation in the mandible.

[Burning mouth]. / Mundbrennen.

BACKGROUND: The complaint of burning mouth is fairly common. Epidemiological studies suggest a prevalence of around 15% in postmenopausal women and between 0,7 and 7,9% for the general population. PURPOSE: The purpose of this article is to present a review of the literature on epidemiology, clinical symptoms, etiology, diagnosis, treatment and prognosis of burning mouth and burning mouth syndrome. RESULTS AND CONCLUSIONS: Potential causal factors have been extensively studied and reviewed, suggesting a multifactorial aetiology, that apparently includes local, systemic and psychogenic factors. However, reports of their relative importance are conflicting. Consequently this lack of evidence is reflected in inconsistent guidelines for diagnosis and treatment. Most of the authors emphasize the importance of history taking and clinical intraoral examination for diagnosis. The symptom of burning mouth associated with clinical mucosal abnormality has to be differentiated from burning mouth syndrome (BMS), a condition in which no mucosal abnormality is evident on examination. Whereas the symptom of burning mouth associated with clinical mucosal signs is described to be often manageable by eliminating possible causal factors, the background of most of the proposed treatments for BMS is empiric or even purely anecdotal. Only a few randomised controlled studies have been performed on treatment outcome. Their differing results are presented in a short overview.

Clp-mediated proteolysis in Gram-positive bacteria is autoregulated by the stability of a repressor.

The heat shock proteins ClpC and ClpP are subunits of an ATP-dependent protease of Bacillus subtilis. Under non-stressed conditions, transcription of the clpC and clpP genes is negatively regulated by CtsR, the global repressor of clp gene expression. Here, CtsR was proven to be a specific substrate of the ClpCP protease under stress conditions. Two proteins of former unknown function, McsA and McsB, which are also encoded by the clpC operon, act as modulators of CtsR repression. McsA containing zinc finger motifs stabilizes CtsR under non-stressed conditions. McsB, a putative kinase, can inactivate CtsR by modification to remove the repressor from the DNA and to target CtsR for degradation by the ClpCP protease during stress. Thus, clp gene expression in Gram-positive bacteria is autoregulated by a novel mechanism of controlled proteolysis, a circuit of down-regulation by stabilization and protection of a transcription repressor, and induction by presenting the repressor to the protease. Thereby, the ClpC ATPase, a member of the Hsp100 family, was identified as a positive regulator of the heat shock response.

The temporomandibular joint and the disc-condyle relationship after functional orthopaedic treatment: a magnetic resonance imaging study.

Causative correction of Class II skeletal malocclusions may be achieved through bite jumping by various means. Numerous animal experiments have yielded evidence of remodelled temporomandibular structures after mandibular protrusion. However, the mode and extent of structural and/or topographic changes of the disc-condyle relationship after functional orthopaedic treatment is still unresolved. A problem exists in defining the physiological position of the condyles and disc-condyle relationship, which is tentatively determined by various methods particularly in magnetic resonance tomographic studies. Despite the high resolution provided, the results have to be interpreted with caution, as osseous resorption and apposition cannot be assessed by visual evidence. This investigation examined the impact on the temporomandibular joints (TMJ), i.e. the condylar shape and position, and the disc-condyle relationship, of the bionator plus extra-oral traction in combination with vertical elastics. The underlying reactions were studied by means of magnetic resonance images (MRI) obtained from n = 15 successfully treated patients (mean age 11.6 years).

Characterisation of the newly identified human Ump1 homologue POMP and analysis of LMP7(beta 5i) incorporation into 20 S proteasomes.

Biogenesis of mammalian 20 S proteasomes occurs via precursor complexes containing alpha and unprocessed beta subunits. A human homologue of the yeast proteasome maturation factor Ump1 was identified in 2D gels of 16 S precursor preparations and designated as POMP (proteasome maturation protein). We show that POMP is detected only in precursor fractions and not in fractions containing mature 20 S proteasome. Northern blot experiments revealed that expression of POMP is induced after treatment with interferon gamma. To analyse the role of the beta 5 propeptide for proper maturation and incorporation of the beta 5 subunit into the complex, human T2 cells, which highly express derivatives of the beta 5i subunit (LMP7), were studied. In contrast to yeast, the presence of the beta 5 propeptide is not essential for incorporation of LMP7 into the proteasome complex. Mutated LMP7 subunits either carrying the prosequence of beta 2i (LMP2) or containing a mutation in the active threonine site are incorporated like wild-type LMP7, while a LMP7 derivative lacking the prosequence completely is incorporated to a lesser extent. Although the absence of the prosequence does not affect incorporation of LMP7, its deletion leads to delayed proteasome maturation and thereby to an accumulation of precursor complexes. As a result of the precursor accumulation, an increased amount of the POMP protein can be detected in these cells.

The clp proteases of Bacillus subtilis are directly involved in degradation of misfolded proteins.

The presence of the heat stress response-related ATPases ClpC and ClpX or the peptidase ClpP in the cell is crucial for tolerance of many forms of stress in Bacillus subtilis. Assays for detection of defects in protein degradation suggest that ClpC, ClpP, and ClpX participate directly in overall proteolysis of misfolded proteins. Turnover rates for abnormal puromycyl peptides are significantly decreased in clpC, clpP, and clpX mutant cells. Electron-dense aggregates, most likely due to the accumulation of misfolded proteins, were noticed in studies of ultrathin cryosections in clpC and clpP mutant cells even under nonstress conditions. In contrast, in the wild type or clpX mutants such aggregates could only be observed after heat shock. This phenomenon supports the assumption that clpC and clpP mutants are deficient in the ability to solubilize or degrade damaged and aggregated proteins, the accumulation of which is toxic for the cell. By using immunogold labeling with antibodies raised against ClpC, ClpP, and ClpX, the Clp proteins were localized in these aggregates, showing that the Clp proteins act at this level in vivo.

A therapeutic concept for the combined orthodontic surgical correction of angle Class II deformities with short-face syndrome: Surgical lengthening of the lower face.

In today's society not only facial esthetics have become important, but also the information on ways to correct adult orthodontic problems is readily available. Subsequently, increasing number of adults seek orthodontic treatment merely to change their facial appearance. In general, these adult patients exhibit such a severe skeletal deformity that it is noticeable even by non-experts. The nature of these adult deformities is such that the only promising treatment is the combined orthodontic-surgical approach. A stable and functional occlusion with a physiologic position for the condyle is the common goal of orthodontic treatment. In patients with skeletal deformities, however, improvement of facial esthetics takes paramount importance. They judge the success of treatment by extraoral appearance. Accordingly, the clinician must assess both the dental and facial appearance, and then inform the patient of different treatment possibilities. In this scenario, patient's input into the decision making process is critical for a mutually satisfactory result. This clinical report describes a concept of systematic approach to treatment of Class II deformities with skeletal deep bite and short lower face (short-face syndrome). This approach emphasizes the soft tissue analysis.

A therapeutic concept for the combined orthodontic surgical correction of angle Class II deformities with short-face syndrome: Surgical lengthening of the lower face.

In today's society not only facial esthetics have become important, but also the information on ways to correct adult orthodontic problems is readily available. Subsequently, increasing number of adults seek orthodontic treatment merely to change their facial appearance. In general, these adult patients exhibit such a severe skeletal deformity that it is noticeable even by non-experts. The nature of these adult deformities is such that the only promising treatment is the combined orthodontic-surgical approach. A stable and functional occlusion with a physiologic position for the condyle is the common goal of orthodontic treatment. In patients with skeletal deformities, however, improvement of facial esthetics takes paramount importance. They judge the success of treatment by extraoral appearance. Accordingly, the clinician must assess both the dental and facial appearance, and then inform the patient of different treatment possibilities. In this scenario, patient's input into the decision making process is critical for a mutually satisfactory result. This clinical report describes a concept of systematic approach to treatment of Class II deformities with skeletal deep bite and short lower face (short-face syndrome). This approach emphasizes the soft tissue analysis.

Effectiveness of intra- and extraoral aids to the bionator. A controlled study within the scope of the "Wuerzburg concept".

A bimaxillary device is effective only if the therapeutic position of the mandible is maintained for as long a period as possible by day and night, i.e. the lower jaw cannot "drop" from the construction bite position. One simple means of preventing the mandible from dropping out of the bimaxillary appliance during sleep and thus of ensuring adaptation of the muscles and of the joint structures during the night without impairing active adaptation during daytime functions (speaking, swallowing) is being used at Wuerzburg University Department of Orthodontics: In connection with anterior traction, which anchors the bimaxillary appliance to the maxilla, attachments are fixed to the mandibular canines or first premolars, so that the mandible is kept in the therapeutic position during the night by means of elastics. Besides the description of this method, the results of a clinical study on its effectiveness are presented in this paper. In the experimental group (20 Class II/1 patients) treated with the combination of bionator with anterior extraoral traction and up-and-down elastics, the increase of the SNB angle and accordingly of the ANB angle was twice as great as in a control group treated only with bionator and extraoral traction without additional securing of the mandible.

Adenovirus death protein, a transmembrane protein encoded in the E3 region, is palmitoylated at the cytoplasmic tail.

The 11.6-K protein of human adenovirus 2 (Ad2), which was recently renamed as adenovirus death protein (ADP), is a type III membrane glycoprotein that ultimately localizes to the nuclear membrane. ADP is encoded in the E3 transcription unit of Ad2 and migrates as a set of multiple bands in SDS-PAGE with three major forms. The corresponding gene product of adenovirus 5 (Ad5) has a slightly lower molecular weight and shows the same pattern in SDS-PAGE. We report here the covalent attachment of fatty acids to cysteine residues of ADP. In the case of Ad5-ADP all three major forms of this protein can be labeled by [3H]palmitic acid, but not by [3H]myristic acid, whereas only two [3H]palmitic acid-labeled Ad2-ADP species could be detected. The label is sensitive to treatment with 1 M hydroxylamine at pH 7 and with 20% beta-mercaptoethanol indicating that the fatty acids are linked via a thioester bond. By thin layer chromatography, the vast majority of the incorporated label was identified as palmitic acid. Two cysteine residues at the boundary between transmembrane domain and cytoplasmic tail which could serve as acceptor sites were mutated to alanine residues by site-directed mutagenesis of the cloned Ad5-ADP gene. Expression of wild-type Ad5-ADP and the resulting mutants was performed in HeLa cells using the vaccinia virus T7 expression system. As demonstrated by labeling with [3H]palmitic acid, only the mutants with one remaining cysteine residue in the cytoplasmic tail were able to incorporate [3H]palmitic acid, indicating that either could serve as acceptor site. In contrast the double cysteine mutant could not be labeled by [3H]palmitic acid, clearly demonstrating that cysteines 53 and 54 are required for palmitoylation and probably represent the palmitoylation sites in Ad5-ADP.

Lüscher colour test in orthodontic patients. Practicability for compliance assessment?

The Lüscher colour test has been repeatedly recommended as a tool for the prediction of orthodontic treatment course and patient cooperation despite its scientific shortcomings. A series of studies involving patients aged 9 to 16 years undergoing treatment with removable appliances at various orthodontic practices and at one university clinic provided no evidence of systematic correlations between Lüscher test colour preferences and cooperation with appliance wear. There was only slight consistency in colour preferences in a re-test conducted a few months after baseline measurements. Statistical evaluation of specific hypotheses and exploratory analysis of the global potential of the test provided no support for generalizable and useful correlations between colour preferences and patient compliance. Therefore the Lüscher test cannot be generally recommended for the rating of patient compliance in the clinical orthodontic practice.

Parental reactions following the birth of a cleft child.

Parental experiences and behaviour following the birth of a cleft child were studied by comparing the results of 2 interview studies conducted at a 7-year interval. The parents of a total of 55 cleft children, most of whom had passed the primary treatment, were interviewed about their psychosocial adaptation, using an interview scheme with the focus on: a) emotional, cognitive and social reactions following birth, b) information-giving and support provided by the medical staff, c) interaction with the cleft child, and d) impact on family life. In both studies, more than 70% of the parents were very shocked at the birth of the cleft child. In the later study, parents more often expressed depression, social avoidance and feelings of guilt. They indicated better psychosocial care and more understanding provided by the medical staff compared to the earlier study. Twice as many parents were satisfied with the information provided by the cleft centre, however, the amount of information had little impact on coping with the shock. In conclusion, further sensitisation of the medical staff with respect to parental coping processes and needs for information and emotional support was called for, along with the inclusion of these topics in the respective curricula.

Effects of information-giving and communication during orthodontic consultation and treatment. Part 3: Optimized orthodontist-patient communication.

Effective, orthodontist-patient communication must meet certain minimum requirements whose importance might vary, depending on the individual case and stage of treatment. In the course of treatment the following tasks are of primary importance: 1. Before the start of treatment, during preliminary consultation, information tailored in extent and quality to the individual patient has to be provided, a professional assessment of treatment needs made, and a relationship of mutual trust established. 2. In the initial phase of treatment, the individual shaping and optimisation of treatment needs is a matter of priority if initial acceptance of the appliance is to be assured. The patient should be advised how to meet the treatment needs in his individual situation. 3. In the further course of treatment, the fulfillment of the demands has to be repeatedly clarified. Feedback with regard to patient cooperation should be handled with caution. A tight recall schedule, a time-keeping record and involvement of the parents are obvious interventions in cases of inadequate cooperation. The treatment may also have to be reevaluated.

Assessment of the combined orthodontic-surgical treatment from the patients' point of view. A longitudinal study.

In a longitudinal study, 40 patients who underwent a combined orthodontic-surgical treatment, were interviewed 4 weeks before operation and 1 week and/or half a year after surgery in order to evaluate patients' expectations and appraisal of the operation and their psycho-social situation. The comparisons between the patients' pre- and postoperative statements yielded the following picture: the patients showed a high degree of satisfaction with the result of the operation. They judged their postoperative facial appearance, with the help of a "semantic differential', to be significantly more attractive. Satisfaction also was expressed in a higher degree of psychological well-being and thus an increase in self-confidence and motivation. Postoperatively, the "Giessen test' showed also a distinctly positive change of the patients' mood and improved experiences with social interaction. Before the operation almost half of the patients feared sensitivity loss or disturbance. One out of 5 patients did in fact suffer from this problem--but not from pain--half a year after the operation. Some patients felt they had received insufficient prior information about the procedure and the risk of the operation. In spite of all burdens for the patients, the assessment of the result of the operation is (generally) positive, not only from the medical point of view.