Ex vivo assessment of the buccal and oral bone by CBCT.

PURPOSE: Identifying buccal and oral bone as an important supporting periodontal structure for teeth provides important information for treatment planning in periodontics and orthodontics. This study aims to add evidence to the knowledge of preciseness of cone beam computed tomography (CBCT) measurements of the vertical dimension of buccal and oral bone. The hypothesis is that CBCT is an accurate and reliable method to measure vertical vestibular and oral bone loss. METHODS: The amount of vertical buccal and oral bone loss (bl) of 260 sites of 10 human cadavers was investigated clinically and radiographically by CBCT. Radiographic measurements were rated by two blinded raters. Measurements and the corresponding differences between clinical and radiological findings are described by medians and quartiles (Q1-Q3). For statistical analysis, Lin's concordance correlation coefficient (CCC) and Bland-Altman plots were calculated. RESULTS: The CCC between the raters was 0.994 (95% confidence interval 0.992-0.995). The median bone loss (bl) distance from the cementoenamel junction (CEJ) to the bony defect (BD) was 3.5 mm (range 3-5 mm). The median bl measured in the CBCT was 3.8 mm (range 3.1-4.8 mm). The median difference of the 2 measurements for all sites included in the study (N = 260) was -0.2 mm (-0.7 to 0.3 mm). CONCLUSIONS: CBCT seems to be an accurate and highly reliable method to detect and describe vertical buccal and oral bone loss. It could improve planning and prediction for successful combined periodontal and orthodontic therapies.

Ex vivo comparison of CBCT and digital periapical radiographs for the quantitative assessment of periodontal defects.

OBJECTIVES: Accurate imaging is essential for effective treatment planning in periodontology. The aim of this ex vivo study was to investigate the accuracy of cone beam computed tomography (CBCT) and digital periapical radiographs (PA) in imaging periodontal defects. Hypotheses are: 1. That CBCT is a more accurate method than PA concerning vertical measurements of periodontal bone defects2. That CBCT itself is an accurate method to describe vertical periodontal bone loss MATERIAL AND METHODS: In this study, 117 periodontal defects from 10 human cadavers were investigated radiographically by CBCT and PA by one calibrated observer. Afterwards the vertical bone loss was measured with a periodontal probe by the same calibrated observer. Differences between radiographic and clinical measurements were calculated and analyzed. Bland-Altmann plots including 95% limits of agreement were calculated. RESULTS: The 95% limits of agreement ranged from 3.29 to -3.27 mm between clinical measurements and measurements in PAs, and from 2.13 to -1.97 mm in CBCTs. The mean difference between clinical and radiographic measurements was 0.0009 mm for PA and 0.0835 mm for CBCT. CONCLUSIONS: When comparing the clinical measurements, CBCT had a higher agreement and less deviations than PAs, and CBCT seems to be an accurate method to describe vertical periodontal bone loss. CLINICAL RELEVANCE: Accurate description of defects is helpful for accurate treatment planning.

[Evaluation of a structured smoking cessation program for primary care medicine]. / Evaluierung eines strukturierten Tabakentwöhnungsprogramms für die ärztliche Praxis.

BACKGROUND: Despite the fact that 9 of 10 general practitioners in Germany believe that smoking cessation is an important topic structured programs are only rarely offered to patients. Beside a lack of time and missing reimbursement, physician's limited treatment skills are frequent reasons for this observation. Therefore we aimed to develop a structured, easy to learn and time-effective smoking cessation program for the general practice. Evidence based treatment elements were combined and standardized by a step by step treatment guideline. METHODS: In a non-interventional observation we tested the program's integration in the daily routines of physicians, the rate of continuous tobacco abstinence after 12 weeks and both patient's and physician's satisfaction with the program and the medication. RESULTS: 44 physicians participated in the study. 184 patients were observed over a mean period of 12.8 weeks and were treated with an individually adapted nicotine replacement therapy for 10 weeks. At the end of treatment (12.1 weeks after the target quit date) 48.4% of the patients reported continuous abstinence. The combination of structured counseling with nicotine substitution in this program was easily implemented in doctor's practice, induced a high user satisfaction and a long usage of medication. The encouraging abstinence rate could have been influenced by selecting highly motivated patients, by offering structured and regular consultations at fixed intervals over the first weeks and by the treatment with the individually adapted nicotine substitution. CONCLUSION: This study shows an easy way to implement an individual and structured smoking cessation therapy in primary care medicine in Germany. Nevertheless, the results should be confirmed in larger cohorts and on a higher methodological level.

New partial sequences of phosphoenolpyruvate carboxylase as molecular phylogenetic markers.

To better understand the evolution of the enzyme phosphoenolpyruvate carboxylase (PEPC) and to test its versatility as a molecular character in phylogenetic and taxonomic studies, we have characterized and compared 70 new partial PEPC nucleotide and amino acid sequences (about 1100 bp of the 3' side of the gene) from 50 plant species (24 species of Bryophyta, 1 of Pteridophyta, and 25 of Spermatophyta). Together with previously published data, the new set of sequences allowed us to construct the up to now most complete phylogenetic tree of PEPC, where the PEPC sequences cluster according to both the taxonomic positions of the donor plants and the assumed specific function of the PEPC isoforms. Altogether, the study further strengthens the view that PEPC sequences can provide interesting information for the reconstruction of phylogenetic relations between organisms and metabolic pathways. To avoid confusion in future discussion, we propose a new nomenclature for the denotation of PEPC isoforms.

Molecular phylogeny of the genus Kalanchoe (Crassulaceae) inferred from nucleotide sequences of the ITS-1 and ITS-2 regions.

The study presents an analysis of genotypic diversity in the genus Kalanchoe (Crassulaceae) on the level of Internal Transcribed Spacer (ITS) sequences and the attempt to correlate this diversity with previous findings on ecophysiological behavior, habitat preference, infrageneric taxonomic position of the species and DNA polymorphism derived from RAPD-PCR data. The Kalanchoe species are mainly abundant in Madagascar and eastern continental Africa and perform in situ diverse modes of crassulacean acid metabolism (CAM), an ecophysiologically relevant adaptation of photosynthesis. Total DNA was extracted from 68 Kalanchoe species and varieties. The ITS-1 and ITS-2 regions of the nuclear RNA genes were amplified by polymerase chain reaction, cloned and sequenced. The alignments of the sequences were evaluated by distance (neighbor joining) and character state (maximum parsimony) methods. The main topologies of the obtained ITS phylogenetic trees were quite similar irrespective of the mode of evaluation and show: (A) within the Crassulaceae the genus Kalanchoe forms a monophyletic clade; and (B) within the genus the species form three main clusters which coincide well with the previously reported three infrageneric sections of the species distinguishable by classical taxonomic criteria, the mode of in situ CAM performance, and DNA fragment pattern obtained by RAPD-PCR analyses. Moreover, the ITS phylogenetic trees show that all African Kalanchoe species form a distinct group within the most derived of the three main clusters. This is consistent with the view that the center of phylogenetic radiation of the genus is located in Madagascar from where the species have spread into the continental Africa.

Identification of phosphoenolpyruvate carboxylase isoforms in leaf, stem and roots of the obligate CAM plant Vanilla planifolia Salib. (Orchidaceae): a physiological and molecular approach.

This study provides the first comparative analysis of phosphoenolpyruvate carboxylase isoforms (PEPc; EC 4.1.1.31) in an obligate crassulacean acid metabolism (CAM) plant, Vanilla planifolia Salisb. (Orchidaceae). Nocturnal CO2 fixation and malate accumulation by the leaves and the green stem show that these organs perform CAM. The chloroplast-containing aerial roots, however, exhibit C3 photosynthesis. The catalytic activity of PEPc was highest in the leaves compared with the stem and aerial roots. The Km (PEP) and Ki (malate) were similar in the PEPc extracted from leaf and aerial roots, and significant higher in stem. cDNA was obtained from those tissues and also from the soil-grown roots, and various cDNA clones were detected and amplified by means of RT-PCR and RACE-PCR. The amino-acid sequences of the PEPc isoforms deduced from the cDNA showed a great degree of homology, and Southern blot analysis suggests that the encoding genes form a small multigene family of at least two members. One PEPc isoform (PpcV1) is assumed to be related to CAM because, as shown by northern blot analysis, it is mainly expressed in the CAM-performing organs, i.e. in the leaves and the stem. A further isoform (PpcV2) was identified in the soil-grown roots and aerial roots, but northern blots show that to some extent PpcV2 is also expressed in the leaf and the stem tissues. Thus, it is assumed that PpcV2 encodes the housekeeping isoform of PEPc. Altogether, the present study provides support in favour of the view that isoforms of PEPc are related to specific functions.

Toward a better knowledge of the molecular evolution of phosphoenolpyruvate carboxylase by comparison of partial cDNA sequences.

To get deeper insight into the evolution of phosphoenolpyruvate carboxylase we have identified PEPC fragments (about 1,100 bp) of another 12 plants species not yet investigated in this context. The selected plants include one Chlorophyta, two Bryophyta, four Pteridophyta, and five Spermatophyta species. The obtained phylogenetic trees on PEPC isoforms are the most complete ones up to now available. Independent of their manner of construction, the resulting dendrograms are very similar and fully consistent with the main topology as it is postulated for the evolution of the higher terrestrial plants. We found a distinct clustering of the PEPC sequences of the prokaryotes, the algae, and the spermatophytes. PEPC isoforms of the archegoniates are located in the phylogenetic trees between the algae and spermatophytes. Our results strengthen the view that the PEPC is a very useful molecular marker with which to visualize phylogenetic trends both on the metabolic and organismic levels.

Chicken microtubule-associated protein 4 (MAP4): a novel member of the MAP4 family.

Chicken gizzard smooth muscle has often been used as a source of proteins of the contractile and cytoskeletal apparatus. In the present study, we isolated a hitherto unknown doublet of proteins, with apparent molecular weights of 200 kDa, from embryonic chicken gizzard and showed its association with the microtubules (MTs) and by immunofluorescence staining of cultured cells. Immunoblot analysis also revealed the ubiquitous expression of this protein in all embryonic chicken tissues examined. Molecular cloning techniques allowed its identification as the chicken homologue of the microtubule-associated protein 4 (MAP4), known from mammalian species, and revealed approximately 90% of its amino acid sequence. MAP4 is the major MAP of non-neuronal tissues and cross-species comparisons clearly demonstrated its highly conserved overall structure, consisting of a basic C-terminal MT-binding region and an acidic N-terminal projection domain of unknown function. Despite these conserved features, overall sequence homologies to its mammalian counterparts are rather low and focused to distinct regions of the molecule. Among these are a conserved 18-amino acid motif, which is known to mediate binding to MTs and a part of the MT-binding domain known as the proline-rich region, which is thought to be the regulatory domain of MAP4. The N-terminal 59 amino acids are a conserved and unique feature of the MAP4 sequence and might be an indication that MAP4 performs other functions besides the enhancement of MT assembly.

Geosiphon pyriforme, a fungus forming endocytobiosis with Nostoc (cyanobacteria), is an ancestral member of the Glomales: evidence by SSU rRNA analysis.

Geosiphon pyriforme inhabiting the surface of humid soils represents the only known example of endocytobiosis between a fungus (Zygomycotina; macrosymbiont) and cyanobacteria (Nostoc; endosymbiont). In order to elucidate the taxonomical and evolutionary relationship of Geosiphon pyriforme to fungi forming arbuscular mycorrhiza (AM fungi), the small-subunit (SSU) ribosomal RNA genes of Geosiphon pyriforme and Glomus versiforme (Glomales; a typical AM fungus) were analyzed and aligned with SSU rRNA sequences of several Basidiomycetes, Ascomycetes, Chytridiomycetes, and Zygomycetes, together with all AM-fungal (Glomales) sequences published yet. The distinct group of the order Glomales, which includes Geosiphon, does not form a clade with any other group of Zygomycetes. Within the Glomales, two main lineages exist. One includes the families Gigasporaceae and Acaulosporaceae; the other one is represented by the genus Glomus, the members of which are very divergent. Glomus etunicatum and Geosiphon pyriforme both form independent lineages ancestral to the Glomales. The data provided by the present paper confirm clearly that Geosiphon represents a fungus belonging to the Glomales. The question remains still open as to whether or not Geosiphon is to be placed within or outside the genus Glomus, since this genus is probably polyphyletic and not well defined yet. Geosiphon shows the ability of a Glomus-like fungus to form a "primitive" symbiosis with a unicellular photoautotrophic organism, in this case a cyanobacterium, leading to the conclusion that a hypothetical association of a Glomus-like fungus with a green alga as a step during the evolution of the land plants appears probable.

Identification of multiple PEPC isogenes in leaves of the facultative Crassulacean acid metabolism (CAM) plant Kalanchoe blossfeldiana Poelln. cv. Tom Thumb.

In the facultative Crassulacean Acid Metabolism (CAM) plant Kulanchoe blossfeldiana cv. Tom Thumb, CAM can be induced by short-day treatment or water deficiency stress. From young leaves of well-watered and water-stressed individuals of this plant, cDNA clones coding for a partial sequence of the key enzyme of CAM, phosphoenolpyruvate carboxylase, were isolated after transcription of mRNA. cDNA polymorphism was established by enzyme restriction profiles and sequencing data. Four PEPC isogenes could be shown to exist in K. blossfeldiana forming two gene pairs, with 95%-98% homology inside and only 75% between the pairs. One cDNA sequence pair having a length of 1113 bp and an open reading frame of 371 AA was identified as PEPC isoform specific for the C3 state, whereas the pair having a length of 1116 bp and an open reading frame of 372 AA could be attributed to the CAM state. These results were confirmed by Southern Blot hybridization.

Complete nucleotide sequence of band 3 related anion transport protein AE2 from human kidney.

The cDNA coding for the complete human band 3 related anion exchange protein AE2 has been cloned from human kidney mRNA. The protein is encoded by a mRNA of approx. 3885 nucleotides containing an open reading frame of 3720 nucleotides. The AE2 protein consists of 1240 amino acid residues and has a molecular mass of 136,805 Da.

[Serological cross-reactions between Echinococcus multilocularis and Dipetalonema viteae infected rodents (author's transl)]. / Serologische Kreuzreaktionen zwischen experimentell mit Echinococcus multiocularis oder Dipetalonema viteae infizierten Nagetieren.

Sera from 30 E. multilocularis infected NMRI-mice and 30 D. viteae infected golden hamsters were tested in the indirect fluorescent antibody (IFAT) and indirect hemagglutination (IHA) tests. In the IFAT there were cross-reactions with all sera, but there were differences between the average titres. While filarial antisera gave higher titres in the homologous system (1:640 against 1:80), it was the opposite with Echinococcus antisera (1:160 against 1:640). Other differences referred to the parasite structures which fluoresced: using Echinococcus antigen the homologous sera gave a bright fluorescence of the outer protoscolex layers including the hooks, while Dipetalonema antisera showed fluorescence of the protoscolex parenchyma only. Thus it is easy to distinguish between homologous and heterologous reactions. Corresponding differences could not be found when using Dipetalonema antigen. In the IHA tests with E. granulosus antigen, which showed an average titre of 1:640 witch E. multilocularis sera, there were few cross-reactions, only 4 out of 30 Dipetalonema antisera giving a positive result.