The British Society of Periodontology referral policy and parameters of care.

This referral policy and parameters of care document was commissioned by the Council of the British Society of Periodontology to provide guidance for practitioners, given the changing environment of primary and secondary care dentistry in the UK. These changes included the provision of the so-called 'high street' specialists in various mono-specialties and engendered a new paradigm of openness and information giving from the profession outwards. This was in turn necessitated by both dento-legal requirements and an increasing demand from the public for the provision of more advanced periodontal care. The promotion of periodontal therapy is both public and practitioner based, and by promoting periodontology to the general practitioner there is an inevitable knock-on effect to the patient base. Thus, in October 2000, these guidelines for referring from primary to secondary care were published so that GDPs could be given some indications of good referral practice, based upon the well-known tenets of the Basic Periodontal Examination (BPE). In addition, a number of succinct and relevant parameters of care were proposed, highlighting the need for the practitioner to not only examine the periodontium but also to make a diagnosis and effect a treatment plan and defined therapeutic goals using evidence based procedures.

Suppression of preadipocyte differentiation and promotion of adipocyte death by HIV protease inhibitors.

Many human immunodeficiency virus (HIV)-infected patients taking combination antiretroviral therapy that includes HIV protease inhibitors experience atrophy of peripheral subcutaneous adipose tissue. We investigated the effects of HIV protease inhibitors on adipogenesis and adipocyte survival using the 3T3-L1 preadipocyte cell line. Several HIV protease inhibitors were found either to inhibit preadipocyte differentiation or to promote adipocyte cell death. One protease inhibitor, nelfinavir, elicited both of these effects strongly. When induced to differentiate in the presence of nelfinavir, 3T3-L1 preadipocytes failed to accumulate cytoplasmic triacylglycerol and failed to express normal levels of the adipogenic transcription factors CCAAT/enhancer-binding protein alpha and peroxisome proliferator-activated receptor gamma. The level of the proteolytically processed, active 68-kDa form of sterol regulatory element-binding protein-1, a transcription factor known to promote lipogenic gene expression, also was reduced markedly in nelfinavir-treated cells, whereas the level of the 125-kDa precursor form of this protein was unaffected. The inhibitory effect of nelfinavir occurred subsequent to critical early events in preadipocyte differentiation, expression of CCAAT/enhancer-binding protein beta and completion of the mitotic clonal expansion phase, because these events were unaffected by nelfinavir treatment. In addition, nelfinavir treatment of fully differentiated 3T3-L1 adipocytes resulted in DNA strand cleavage and severe loss of cell viability. In contrast, cell proliferation and viability of preadipocytes were unaffected by nelfinavir treatment. Thus, molecular or cellular changes that occur during acquisition of the adipocyte phenotype promote susceptibility to nelfinavir-induced cell death. When considered together, these results suggest that nelfinavir may promote adipose tissue atrophy by compromising adipocyte viability and preventing replacement of lost adipocytes by inhibiting preadipocyte differentiation.

Interaction of GRASP, a protein encoded by a novel retinoic acid-induced gene, with members of the cytohesin family of guanine nucleotide exchange factors.

A novel, retinoic acid-induced gene, GRP1-associated scaffold protein (GRASP), was isolated from P19 embryonal carcinoma cells using a subtractive screening strategy. GRASP was found to be highly expressed in brain and exhibited lower levels of expression in lung, heart, embryo, kidney, and ovary. The predicted amino acid sequence of GRASP is characterized by several putative protein-protein interaction motifs, suggesting that GRASP may be a component of a larger protein complex in the cell. Although GRASP does not harbor a predicted membrane spanning domain(s), the protein was observed to be associated with the plasma membrane of transiently transfected mammalian cells. Yeast two-hybrid screening revealed that GRASP interacted strongly with the General Receptor for Phosphoinositides 1 (GRP1), a brefeldin A-insensitive guanine nucleotide exchange factor for the ADP-ribosylation factor family of proteins. GRASP. GRP1 interactions were also demonstrated in vitro and in mammalian cells in which GRASP was shown to enhance GRP1 association with the plasma membrane. Furthermore, GRASP colocalized with endogenous ADP-ribosylation factors at the plasma membrane in transfected cells, suggesting that GRASP may modulate signaling by this family of small GTPases.

Management of molar teeth with periodontal disease.

Over the years patients' attitudes towards maintaining a functional and aesthetic masticatory apparatus have improved, and their expectations of delivery of care by the dental professional have risen. With the advance of new techniques and materials, the periodontist can now offer an ever-expanding range of treatments in the management of molar teeth with periodontal disease. This paper considers such treatment in relation to the levels of disease present and within the overall context of adult restorative dental healthcare.

The human p53 negative regulatory domain mediates inhibition of reporter gene transactivation in yeast lacking thioredoxin reductase.

Stimulation of target gene transcription by human p53 is inhibited in budding yeast lacking the TRR1 gene encoding thioredoxin reductase. LexA/p53 fusion proteins were used to study the basis for thioredoxin reductase dependence. A fusion protein containing all 393 of the residues of p53 efficiently and specifically stimulated transcription of a LexOP-LacZ reporter gene in wild-type yeast but was several-fold less effective in delta trr1 yeast lacking the thioredoxin reductase gene. Thus, even when p53 was tethered to a reporter gene by a heterologous DNA-binding domain, reporter gene transactivation remained dependent on thioredoxin reductase. A fusion protein containing only the activation domain of p53 stimulated reporter gene transcription equally in wild-type and delta trr1 cells, suggesting that p53 residues downstream from the activation domain created the requirement for thioredoxin reductase. Experiments using additional LexA/p53 truncation mutations indicated that the p53 negative regulatory domain, rather than the DNA-binding or oligomerization domains, created the requirement for thioredoxin reductase. The fusion protein results suggested that, under oxidative conditions, the negative regulatory domain inhibited the ability of DNA-bound p53 to stimulate transcription. However, deletion of the negative regulatory domain did not alleviate the requirement of non-LexA-containing p53 for thioredoxin reductase. The results, thus, suggest that oxidative conditions inhibit both DNA binding and transactivation by p53, and that inhibition of the latter requires the negative regulatory domain.

Heterodimeric interactions between chicken ovalbumin upstream promoter-transcription factor family members ARP1 and ear2.

Members of the chicken ovalbumin upstream promoter-transcription factor (COUP-TF) subfamily of orphan nuclear receptors, which minimally includes COUP-TFI and ARP1, are highly expressed in brain and are generally considered to be constitutive repressors of transcription. We have used a yeast two-hybrid system to isolate proteins expressed in brain that interact with ARP1. One of the proteins isolated in this screen was Ear2, another orphan receptor that has been suggested to be a member of the COUP-TF subfamily. Here we demonstrate that ARP1 and Ear2 form heterodimers in solution and on directly repeated response elements with high efficiency and a specificity differing from that of homodimeric complexes composed of either receptor. ARP1 and Ear2 were observed to interact in mammalian cells, and the tissue distribution of Ear2 transcripts was found to overlap precisely with the expression pattern of ARP1 in several mouse tissues and embryonal carcinoma cell lines. Heterodimeric interactions between ARP1 and Ear2 may define a distinct pathway of orphan receptor signaling.

Identification of nuclear receptor corepressor as a peroxisome proliferator-activated receptor alpha interacting protein.

Nuclear receptor corepressor (NCoR) was demonstrated to interact strongly with peroxisome proliferator-activated receptor alpha (PPARalpha), and PPARalpha ligands suppressed this interaction. In contrast to the interaction of PPARalpha with the coactivator protein, p300, association of the receptor with NCoR did not require any part of the PPARalpha ligand binding domain. NCoR was found to suppress PPARalpha-dependent transcriptional activation in the context of a PPARalpha.retinoid X receptor alpha (RXRalpha) heterodimeric complex bound to a peroxisome proliferator-responsive element in human embryonic kidney 293 cells. This repression was reversed agonists of either receptor demonstrating a functional interaction between NCoR and PPARalpha.RXRalpha heterodimeric complexes in mammalian cells. NCoR appears to influence PPARalpha signaling pathways and, therefore, may modulate tissue responsiveness to peroxisome proliferators.

Ligand-induced peroxisome proliferator-activated receptor alpha conformational change.

Structurally diverse peroxisome proliferators and related compounds that have been demonstrated to induce the ligand-dependent transcriptional activation function of mouse peroxisome proliferator-activated receptor alpha (mPPARalpha) in transfection experiments were tested for the ability to induce conformational changes within mPPARalpha in vitro. WY-14,643, 5,8,11,14-eicosatetraynoic acid, LY-171883, and clofibric acid all directly induced mPPARalpha conformational changes as evidenced by a differential protease sensitivity assay. Carboxyl-terminal truncation mutagenesis of mPPARalpha differentially affected the ability of these ligands to induce conformational changes suggesting that PPAR ligands may make distinct contacts with the receptor. Direct interaction of peroxisome proliferators and related compounds with, and the resulting conformational alteration(s) in, mPPARalpha may facilitate interaction of the receptor with transcriptional intermediary factors and/or the general transcription machinery and, thus, may underlie the molecular basis of ligand-dependent transcriptional activation mediated by mPPARalpha.

p300 functions as a coactivator for the peroxisome proliferator-activated receptor alpha.

The integrator protein, p300, was demonstrated to interact with mouse peroxisome proliferator-activated receptor alpha in a ligand-enhanced manner. The PPARalpha-interacting domain of p300 was mapped to amino acids 39-117 which interacted strongly with PPARalpha but did not interact with retinoic acid receptor-gamma or retinoid X receptor-alpha. Amino acids within the carboxyl terminus of PPARalpha as well as residues within the hinge region were required for ligand-dependent interaction with p300. p300 enhanced the transcriptional activation properties of PPARalpha and, therefore, can be considered a bona fide coactivator for this nuclear receptor. These observations extend the group of p300-interacting proteins to include mPPARalpha and further characterize the molecular mechanisms of PPARalpha-mediated transcriptional regulation.

A comparative clinical study: the use of human type I collagen with and without the addition of metronidazole in the GTR method of treatment of periodontal disease.

This investigation was undertaken to evaluate cross-linked human type I collagen, with and without added metronidazole, when used as a barrier membrane in the guided tissue regeneration (GTR) principle of treatment for periodontal disease. 16 patients suffering from moderate to severe periodontitis with 78 bilaterally matched periodontal defects underwent similar contralateral surgical flap procedures after preliminary scaling, polishing and oral hygiene instruction. At the experimental sites, which were selected at random, the flap was closed over metronidazole impregnated collagen as a GTR membrane, the contralateral sites receiving a plain collagen barrier as control. The plaque index (PLI), gingival index (GI), bleeding index (BI), probing pocket depth (PPD) and probing attachment level (PAL) were recorded at baseline, 6, 12 and 26 weeks post-operatively. The bony defects were classified and furcation involvement noted. The clinical parameters were recorded by an examiner, other than the surgeon, who had been previously assessed for accurate reproducibility of measurements and was unaware of the experimental sites. PPD and PAL were measured with a constant pressure probe, localised by a soft stent. Post-operative discomfort was evaluated by means of a questionnaire. PLI, GI and BI were significantly improved compared to baseline for both test and control sites at 6, 12 and 26 weeks post surgery (p < 0.001) but there was no significant difference between these sites (p > 0.05). There was a reduction in PPD at 6 weeks which was significant at 12 and 26 weeks post-operatively (p < 0.001) for both test and control sites, but no difference between these sites was evident (p > 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Clinical trial of cross-linked human type I collagen as a barrier material in surgical periodontal treatment.

This study was undertaken to evaluate freeze-dried cross-linked human type I collagen when used as a barrier membrane for guided tissue regeneration (GTR) in periodontal surgery. 14 patients with radiographic evidence of bone loss and residual pocketing of > 6 mm on bilaterally matched sites were given preliminary scaling, polishing and oral hygiene instruction before undergoing contralateral flap surgery. At the experimental sites, a collagen membrane was adapted to the root surfaces, extending from 2 mm apical to the bone crest to just subgingival, before replacing the flap and closing with sutures. The control sites underwent a similar procedure but without the placement of the collagen barrier. The experimental sites were selected at random. Plaque index (PLI), gingival index (GI), bleeding index (BI), probing pocket depth (PPD) and probing attachment level (PAL) were recorded at baseline, 6, 12 and 26 weeks post-operatively. The bony defects were classified and furcation involvement noted. The clinical parameters were recorded by an examiner, previously assessed for accurate reproducibility of measurement, who was not the surgeon and unaware of the experimental sites. PPD and PAL were measured using a constant pressure probe localised by a soft stent. There was significant improvement in the PLI, GI and BI at both test and control sites at 6, 12 and 26 weeks compared to baseline (P < 0.001 for PLI; P < 0.0001 for GI and BI) but not significant difference between these sites.(ABSTRACT TRUNCATED AT 250 WORDS)

The pharmacokinetics of suriclone after single- and multiple-dose administration in healthy elderly volunteers.

Healthy elderly people aged 70 or more should receive a starting dosage of the anxiolytic drug suriclone half that recommended for younger people; frail or ill elderly people may require further dosage reduction. These conclusions were based on a study of the pharmacokinetics and tolerability of orally administered suriclone after a single dose of 0.2mg and after multiple doses of 0.2mg 3-times daily in 8 male and 8 female healthy elderly volunteers. Informal comparisons of the pharmacokinetic data from this study with studies in younger volunteers showed a 53% decrease in clearance and an 84% increase in elimination half-life. Area under the plasma concentration-time curve (AUC) for suriclone and total antigenic products in the elderly were twice the values in younger volunteers when adjusted for different dosage. There were no significant changes in pulse rate or blood pressure during the study. One person showed a change from sinus rhythm to atrial fibrillation. Six people had transient adverse events including unsteadiness, vomiting, difficulty in reaching for objects, urinary incontinence and sedation.

Evaluation of functional analogs of CC-1065 and the duocarmycins incorporating the cross-linking 9a-chloromethyl-1,2,9,9a-tetrahydrocyclopropa[c]benz[e]indol-4-on e (C2BI) alkylation subunit.

The DNA alkylation properties and in vitro cytotoxic activity of a series of analogs of CC-1065 and the duocarmycins incorporating the 9a-chloromethyl-1,2,9,9a-tetrahydrocyclopropa[c]benz[e]indol-4-one (C2BI) alkylation subunit are detailed. The C2BI-based agents have been shown to alkylate DNA within the minor groove in a fashion analogous to CC-1065 or duocarmycin. The stereoelectronically-controlled adenine N3 addition to the least substituted cyclopropane carbon occurs with a selectivity that represents a composite of the two enantiomers of the corresponding CBI-based agents. Additional high affinity alkylation sites were detected which were not prominent alkylation sites for either enantiomer of the CBI-based agents. Such sites may represent induced high affinity alkylation sites resulting from DNA cross-linking following complementary strand alkylation at a high affinity alkylation site and each such site detected proved consistent with predicted models of an adenine-adenine cross-linking event. Further, consistent with this interpretation, the C2BI agents were shown to constitute efficient cross-linking agents with DNA cross-linking being observed at the same concentrations as DNA alkylation. In comparison to the parent CBI-based agents, the C2BI-based agents proved to be approximately 100-10,000x less effective at DNA alkylation and 100-10,000x less potent in cytotoxic assays. This is suggested to be the consequence of a significant steric deceleration of the adenine N3 alkylation reaction attributable to the additional 9a-chloromethyl substituent. Consistent with this interpretation, the noncovalent binding constant of C2BI-CDPI2 for poly[dA]-poly[dA]-poly[dT] proved nearly identical to that of CDPI3 under kinetic binding conditions, and prolonged incubation of C2BI-CDPI2 with poly[dA]-poly[dT] (72 h, 25 degrees C) provided covalent complexes with a helix stabilization comparable to that observed with (+)- or (-)-CPI-CDPI2 indicating that the size of the C2BI subunit inhibits but does not preclude productive DNA alkylation.

Freeze-dried, cross-linked bovine type I collagen: analysis of properties.

This study was undertaken to assess the physical and biological properties of freeze-dried cross-linked bovine type I collagen and to assess its potential for use in the guided tissue regeneration method of treatment of periodontal disease in human adult subjects. The modulus of elasticity, swelling ratio, and biodegradation rate were investigated. The collagen sponge was implanted subdermally into Sprague-Dawley rats and a histological study carried out at 2, 7, 21, 35, and 49 days post implantation. Growth of human gingival and periodontal ligament derived fibroblasts on collagen sponge was assessed, as well as the effect of bovine collagen supernatants upon gingival and periodontal fibroblast cultures. The physical properties of the collagen sponge were consistent with good handling qualities and, therefore, it was appropriate for use at a surgical site. The histological study demonstrated a reduction in thickness of the collagen at 21 days; at 35 days there was a hazy appearance of the collagen remnants; and at 49 days the graft material had been completely replaced with fibrous tissues. The in vitro response of human gingival and periodontal fibroblasts to bovine collagen showed that, after 21 days, confluent fibroblast growth was observed around and underneath the sponge. The effect of bovine collagen supernatants upon fibroblasts demonstrated an apparent proliferative effect of the supernatant with both gingival and periodontal ligament fibroblasts. However, the non-parametric Friedman test revealed no significant differences between dilutions or time points. The overall findings provide encouraging evidence of the safety of freeze-dried cross-linked bovine collagen sponge in the surgical treatment of periodontal disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Guided tissue regeneration.

One of the main goals of periodontal therapy is the predictable regeneration of the periodontium by allowing repopulation of periodontal ligament cells into the wound area after surgery and preventing the colonisation of the exposed root surface with epithelial, gingival, and bone cells. In order to achieve this, emphasis has been placed on the use of barrier materials in the form of semipermeable membranes which are interposed between the mucoperiosteal flap and the bone and tooth surfaces during surgery. This technique is known as 'guided tissue regeneration' (GTR) and this article looks at the theory and practice of GTR, as well as reviewing the use of non-resorbable membrane materials such as expanded polytetrafluor-ethylene and ethyl cellulose, and resorbable materials such as collagen and polylactic acid.

Dentine hypersensitivity: effect of interactions between metal salts, fluoride and chlorhexidine on the uptake by dentine.

A large number of compounds may be applied to the teeth for preventive or therapeutic purposes, most notably in dentine hypersensitivity. The uptake of any one compound by tooth substances may clearly be affected by another to produce synergistic, additive or antagonistic effects. This study determined whether uptake interactions occurred between fluoride, chlorhexidine, strontium, tin and zinc. All compounds, in particular tin, showed considerable affinity for dentine. Fluoride reduced the uptake of zinc but not tin, strontium or chlorhexidine. Chlorhexidine increased the uptake of zinc, but not tin, strontium or fluoride. Zinc and tin decreased fluoride uptake but strontium had no such effect. Finally, chlorhexidine uptake was increased by zinc and tin but not by fluoride or strontium. Based on the theories proposed for the reaction of these various compounds with calcified tissues, explanations for the interactions can be made. The results appear relevant to some reported clinical effects of combinations of these compounds and indicate that such interactions may have advantageous or disadvantageous consequences depending on the basis for treatment.

The pharmacokinetics of fentiazac and its metabolite, p-hydroxyfentiazac, after twice-daily oral administration to male volunteers.

The pharmacokinetics of the anti-inflammatory agent fentiazac and its principal plasma metabolite, p-hydroxyfentiazac, have been investigated after repeated oral administration of fentiazac to male volunteers. Each volunteer received 200 mg of fentiazac twice daily for 15 d. Absorption was quite rapid, though some inter-subject variation in rates of absorption and bioavailability was observed. tmax values after the first dose ranged from 0.75-3 h while Cpmax values were 1050-4880 ng/ml. Elimination of fentiazac from plasma occurred rapidly in curvilinear fashion, so that concentrations were only 1% of their maximum value by 12 h after dosing. Maximum concentrations of p-hydroxyfentiazac after a single dose of fentiazac were 25.6-79.4% of those of fentiazac and were achieved at similar times. The metabolite was more slowly eliminated; the mean concentration of p-hydroxyfentiazac 12 h after a single dose was still 8% of its maximal value. On repeated administration, AUC0-12 h values for fentiazac and hydroxyfentiazac increased, as indicated by accumulation factors of 1.17 +/- 0.11 and 1.30 +/- 0.11 on days 8 and 15, respectively, for fentiazac and 1.72 +/- 0.15 and 1.77 +/- 0.10 for hydroxyfentiazac. There was no significant difference between days 8 and 15 in the extent of accumulation of either compound. Trough concentrations of fentiazac and hydroxyfentiazac on days 12 and 15 were similar to those on day 8. The clinical significance of these observations is discussed.