Dentine sialophosphoprotein signal in dentineogenesis and dentine regeneration.

Dentineogenesis starts on odontoblasts, which synthesise and secrete non-collagenous proteins (NCPs) and collagen. When dentine is injured, dental pulp progenitors/mesenchymal stem cells (MSCs) can migrate to the injured area, differentiate into odontoblasts and facilitate formation of reactionary dentine. Dental pulp progenitor cell/MSC differentiation is controlled at given niches. Among dental NCPs, dentine sialophosphoprotein (DSPP) is a member of the small integrin-binding ligand N-linked glycoprotein (SIBLING) family, whose members share common biochemical characteristics such as an Arg-Gly-Asp (RGD) motif. DSPP expression is cell- and tissue-specific and highly seen in odontoblasts and dentine. DSPP mutations cause hereditary dentine diseases. DSPP is catalysed into dentine glycoprotein (DGP)/sialoprotein (DSP) and phosphoprotein (DPP) by proteolysis. DSP is further processed towards active molecules. DPP contains an RGD motif and abundant Ser-Asp/Asp-Ser repeat regions. DPP-RGD motif binds to integrin αVß3 and activates intracellular signalling via mitogen-activated protein kinase (MAPK) and focal adhesion kinase (FAK)-ERK pathways. Unlike other SIBLING proteins, DPP lacks the RGD motif in some species. However, DPP Ser-Asp/Asp-Ser repeat regions bind to calcium-phosphate deposits and promote hydroxyapatite crystal growth and mineralisation via calmodulin-dependent protein kinase II (CaMKII) cascades. DSP lacks the RGD site but contains signal peptides. The tripeptides of the signal domains interact with cargo receptors within the endoplasmic reticulum that facilitate transport of DSPP from the endoplasmic reticulum to the extracellular matrix. Furthermore, the middle- and COOH-terminal regions of DSP bind to cellular membrane receptors, integrin ß6 and occludin, inducing cell differentiation. The present review may shed light on DSPP roles during odontogenesis.

The silencing of cathepsin K used in gene therapy for periodontal disease reveals the role of cathepsin K in chronic infection and inflammation.

BACKGROUND AND OBJECTIVE: Periodontitis is a severe chronic inflammatory disease and one of the most prevalent non-communicable chronic diseases that affects the majority of the world's adult population. While great efforts have been devoted toward understanding the pathogenesis of periodontitis, there remains a pressing need for developing potent therapeutic strategies for targeting this dreadful disease. In this study, we utilized adeno-associated virus (AAV) expressing cathepsin K (Ctsk) small hairpin (sh)RNA (AAV-sh-Ctsk) to silence Ctsk in vivo and subsequently evaluated its impact in periodontitis as a potential therapeutic strategy for this disease. MATERIAL AND METHODS: We used a known mouse model of periodontitis, in which wild-type BALB/cJ mice were infected with Porphyromonas gingivalis W50 in the maxillary and mandibular periodontium to induce the disease. AAV-sh-Ctsk was then administrated locally into the periodontal tissues in vivo, followed by analyses to assess progression of the disease. RESULTS: AAV-mediated Ctsk silencing drastically protected mice (> 80%) from P. gingivalis-induced bone resorption by osteoclasts. In addition, AAV-sh-Ctsk administration drastically reduced inflammation by impacting the expression of many inflammatory cytokines as well as T-cell and dendritic cell numbers in periodontal lesions. CONCLUSION: AAV-mediated Ctsk silencing can simultaneously target both the inflammation and bone resorption associated with periodontitis through its inhibitory effect on immune cells and osteoclast function. Thereby, AAV-sh-Ctsk administration can efficiently protect against periodontal tissue damage and alveolar bone loss, establishing this AAV-mediated local silencing of Ctsk as an important therapeutic strategy for effectively treating periodontal disease.

Impact of Periodontal Disease on the Quality of Life of Older People in Indonesia: A Qualitative Study.

INTRODUCTION: The impact of periodontal disease on oral health-related quality of life (OHRQoL) has often been investigated from a quantitative research perspective, which is based on clinical findings and an OHRQoL questionnaire. Very few studies have examined the issue from the view of qualitative research. To our knowledge, there have been no previous qualitative studies focusing the effect of periodontal disease on OHRQoL in Indonesian older people. OBJECTIVES: To explore and understand the impact of periodontal disease on the OHRQoL of older people as a subjective reflection in relation to periodontal disease experiences. METHODS: Semi-structured interviews were conducted in a sample of 31 older people with generalized chronic periodontitis. Thematic analysis was used to identify the key issues in participants' accounts. The analysis was undertaken by 2 independent coders to ensure reliability. To achieve thematic saturation, successive interviews were undertaken until 5 sequential interviews did not bring new themes. RESULTS: Participants reported the negative effects likely related to periodontal disease. The impacts of periodontal disease were described by these older people as affecting more than pain, physical discomfort, and physical function restrictions. Periodontal disease also affected their psychological and social aspects of daily living. In addition, this study identified themes related to individual and environmental factors that may modify and personalize periodontal disease experiences. Furthermore, this study identified a misleading belief that problems related to periodontal disease were a normal part of aging, which might influence individuals' expectations toward oral health. Relatedly, participants frequently reported that the progression of tooth mobility to tooth loss was an inevitable part of the aging process. CONCLUSIONS: Periodontal disease negatively affected participants' OHRQoL. It is fundamental to understand older people's perceptions toward their periodontal disease as well as individual and environmental factors that may have an influence on their periodontal disease experiences. KNOWLEDGE TRANSFER STATEMENT: This study is a reflection of Indonesian older people's subjective periodontal disease experiences. Therefore, the present study can be used to understand older people's perceptions, attitudes, behaviors, and experiences toward periodontal disease and how this disease may affect their quality of life. This study also highlights a widespread and misleading belief that oral problems related to periodontal disease are an inevitable part of aging in this study population.

Periodontal Disease and Oral Health-Related Quality of Life in the Older Population in Indonesia.

INTRODUCTION: Despite being acknowledged as the second global burden of oral disease, periodontal disease has few epidemiologic studies in the literature, particularly for developing countries. Many previous studies have assessed the relationship between periodontal disease and oral health-related quality of life (OHRQoL), with patients attending dental clinic or hospitals rather than a general population. This study attempted to fill the knowledge gap in limited information about periodontal disease and OHRQoL, with reference to a general population in a developing country. OBJECTIVES: To investigate the relationship between OHRQoL and periodontal diseases in an older population in Indonesia. METHODS: We invited 582 older people from community health centers. The 369 (63.4%) older people who agreed to participate consented to an oral health examination and a questionnaire capturing demographic, socioeconomic, behavioral, and Oral Health Impact Profile-14 (OHIP-14) data. RESULTS: Almost 75% of the older people had generalized periodontitis; 3% had healthy periodontal status; and around 22% had localized periodontitis. There was a lack of statistical evidence for an association between periodontal disease status and OHRQoL. This result was based on the appraisal of the prevalence of the impact (Odds ratio [OR], 0.95 [95% CI, 0.54 to 1.59]; P = 0.77), difference in mean severities (0.07 [95% CI, -1.66 to 1.80]; P = 0.94), and extent of the impact (P = 0.996). However, we found evidence for a relationship between tooth mobility and OHRQoL for all of the OHIP assessments, including prevalence of the impact (OR, 1.87 [95% CI, 1.16 to 3.01]; P = 0.009), difference in mean severities (-2.98 [95% CI, -4.50 to -1.45]; P < 0.001), and extent of the impact (P = 0.001). CONCLUSION: There was a lack of statistical evidence for a relationship between periodontal disease status and OHRQoL in this society. However, we found evidence that tooth mobility, as a sign of periodontal disease progression, is related to OHRQoL. KNOWLEDGE TRANSFER STATEMENT: The present study can be used by dentists, community health workers, and policy makers in Indonesia to understand the prevalence, severity, and extent of the negative impacts of periodontal disease on older people's quality of life. In addition, this study provides information about factors that might considerably affect the oral health-related quality of life in this society, such as brushing habits, dental visit, family income, DMF-T status, and subjective appraisal toward dental health.

Differential enamel and osteogenic gene expression profiles in odontogenic tumors.

Odontogenic tumors occur within the jaw bones and may be derived from odontogenic epithelium or ectomesenchyme or contain active components of both tissue types. We investigated the gene expression profile of enamel matrix proteins (EMPs), genes related to osteogenesis, and the mineralization process in odontogenic tumor cell populations focusing on an ameloblastoma (AB-1), a keratocystic odontogenic tumor (KCOT-1), and a calcifying epithelial odontogenic tumor (CEOT-1). All cell populations were shown to be epithelial in origin by CK14 expression. All tested EMPs were expressed by all odontogenic tumor cell types, with higher transcript levels seen in the AB-1 population especially for AMEL, AMBN, and ODAM. CEOT-1 cell populations showed a greater content of ALP-positive cells as well as higher ALP mRNA levels. Using qRT-PCR, we found a higher expression of 8 genes in the CEOT-1 compared to the AB-1 and KCOT-1. In this study we demonstrated the establishment of AB-1, KCOT-1 and CEOT-1 cell populations. The unique gene expression profiles of AB-1, KCOT-1, and CEOT-1 cells and their interactions with the surrounding microenvironment may support their unique tumor development, progression, and survival.

Prognostic significance of the lung scintigraphy scan result and corresponding chest X-ray in patients with suspected pulmonary embolism.

AIM: To determine whether the survival of patients with suspected acute pulmonary embolism (PE) relates to radiological probability of acute PE assessed using lung scintigraphy scans (LSS). METHODS: Lung scintigraphy scan results from a venous thromboembolism database were categorised as high, indeterminate or low probability using the modified PIOPED criteria and corresponding chest X-rays (CXRs) as normal or abnormal. Mortality data on these cases were obtained from the General Register Office for Scotland, and survival was analysed using the Kaplan-Meier method. RESULTS: Of the 1,818 LSS analysed, 941 (51.8%) were normal, 532 (29.3%) indeterminate and 345 (19.0%) high probability. After an adjustment for age and gender, no significant survival difference was found between patients with normal and high probability LSS (p=0.182). However, patients with indeterminate LSS had significantly lower survival than patients in the other groups. This difference persisted after adjustment for CXR result. CONCLUSIONS: Indeterminate LSS results are associated with a poor prognosis. Careful follow-up of patients with inderminate LSS would appear to be justified.

Phenotype characterization and DSPP mutational analysis of three Brazilian dentinogenesis imperfecta type II families.

The aim of this study was to perform phenotype analysis and dentin sialophosphoprotein (DSPP) mutational analysis on 3 Brazilian families diagnosed with dentinogenesis imperfecta type II (DGI-II) attending the Dental Anomalies Clinic in Brasilia, Brazil. Physical and oral examinations, as well as radiographic and histopathological analyses, were performed on 28 affected and unaffected individuals. Clinical, radiographic and histopathological analyses confirmed the diagnosis of DGI-II in 19 individuals. Pulp stones were observed in ground sections of several teeth in 2 families, suggesting that obliteration of pulp chambers and root canals results from the growth of these nodular structures. Mutational DSPP gene analysis of representative affected family members revealed 7 various non-disease-causing alterations in exons 1-4 within the dentin sialoprotein domain. Further longitudinal studies are necessary to elucidate the progression of pulpal obliteration in the DGI-II patients studied as well as the molecular basis of their disease.

Anti-Saccharomyces cerevisiae antibodies status is associated with oral involvement and disease severity in Crohn disease.

OBJECTIVES: To determine anti-Saccharomyces cerevisiae antibodies (ASCA) status and its relation to disease phenotype in patients with inflammatory bowel disease (IBD). PATIENTS AND METHODS: A total of 301 Scottish patients with early-onset IBD-197 Crohn disease (CD), 76 ulcerative colitis (UC), 28 indeterminate colitis (IC)-and 78 healthy control individuals were studied. ASCA status (IgA, IgG) was determined by enzyme-linked immunosorbent assay. ASCA status was then analyzed in relation to CD phenotype. RESULTS: Patients with CD had a higher prevalence of ASCA than patients with UC and healthy controls: 82/197 versus 12/76, odds ratio (OR) 3.80 (1.93-7.50) and 82/197 versus 6/78, OR 8.56 (3.55-20.62), respectively. Univariate analysis showed that positive ASCA status was associated with oral CD (17/25 vs 59/153, OR 3.39 [1.38-8.34]), perianal CD (39/77 vs 38/108, OR 1.89 [1.04-3.44]) and the presence of granulomata (63/132 vs 15/52, OR 2.25 [1.13-4.48]) and also with markers of disease severity: raised C-reactive protein (44/90 vs 12/49, OR 2.95[1.36-6.37]), hypoalbuminemia (44/85 vs 20/74, OR 2.28[1.19-4.37]), and surgery (27/49 vs 54/147, OR 2.11 [1.10-4.06]). From multivariate analysis, the presence of oral disease (adjusted P = 0.001, OR 22.22 [3.41-142.86]) and hypoalbuminemia (adjusted P = 0.01, OR 4.78 [1.40-16.39]) was found to be independently associated with ASCA status. No association was demonstrated between ASCA and IBD candidate genes. CONCLUSIONS: Patients with CD had a higher prevalence of ASCA than did other patients with IBD. ASCA status described patients with CD who had a specific phenotype, showing an association with markers of disease severity and oral CD involvement.

General practitioner involvement and patient outcomes in HIV management.

We undertook this study to try to determine whether disease outcomes were poorer in patients with HIV infection whose general practitioner (GP) was unaware of their status compared with those whose GP was aware. The notes of 375 HIV-positive patients attending Edinburgh's genitourinary (GU) medicine clinic were reviewed. The GPs of 292 patients (78%) had been informed of their patient's HIV infection. Advancing disease was associated with disclosure of the status to GPs (P = 0.037) but no significant association was found between informing GPs and the viral load results of treated (P = 0.389) and untreated patients (P = 0.070). Twenty-three percent of patients had had one or more bacterial sexually transmitted infections (STIs) while receiving their HIV care at a GU medicine clinic. Patients diagnosed with an STI were less likely to disclose their HIV status to their GP (P < 0.0005). Non-disclosure of the HIV status to a GP may be a predictor of unsafe sexual practices.

Third Dentition Is the Main Cause of Premolar Supernumerary Tooth Formation.

While the prevalence of supernumerary teeth (ST) is high in permanent dentition, the etiology of ST in humans remains unclear. However, multiple murine models of ST have elaborated on dated mechanisms traditionally ascribed to ST etiology: one involves the rescue of rudimental teeth, and the second considers the contribution of odontogenic epithelial stem cells. It remains unclear whether these mechanisms of ST formation in mice are applicable to humans. The third dentition is usually regressed apoptotic-that is, the teeth do not completely form in humans. Recently, it was suggested that ST result from the rescue of regression of the third dentition in humans. The present investigation evaluates the proportion of collected general ST cases that evinced a third dentition based on the clinical definition of ST derived from the third dentition. We also investigated the contribution of SOX2-positive odontogenic epithelial stem cells to ST formation in humans. We collected 215 general ST cases from 15,008 patients. We confirmed that the general characteristics of the collected ST cases were similar to the results from previous reports. Of the 215 cases, we narrowed our analysis to the 78 patients who had received a computed tomography scan. The frequency of ST considered to have been derived from the third dentition was 26 out of 78 cases. Evidence of a third dentition was especially apparent in the premolar region, was more common in men, and was more likely among patients with ≥3 ST. SOX2-positive odontogenic epithelial stem cells within the surrounding epithelial cells of developing ST were observed in non-third dentition cases and not in third dentition cases. In conclusion, the third dentition is the main cause of ST in humans. The odontogenic epithelial stem cells may contribute to ST formation in cases not caused by a third dentition.

PDGF up-regulates CSF-1 gene transcription in ameloblast-like cells.

Macrophage colony-stimulating factor (CSF-1) is a key regulatory cytokine for amelogenesis, and ameloblasts synthesize CSF-1. We hypothesized that PDGF stimulates DNA synthesis and regulates CSF-1 in these cells. We determined the effect of PDGF on CSF-1 expression using MEOE-3M ameloblasts as a model. By RT-PCR, MEOE-3M expressed PDGFRs and PDGF A- and B-chain mRNAs. PDGF-BB increased DNA synthesis and up-regulated CSF-1 mRNA and protein in MEOE-3M. Cells transfected with CSF-1 promoter deletion constructs were analyzed. A PDGF-responsive region between -1.7 and -0.795 kb, containing a consensus Pea3 binding motif, was identified. Electrophoretic mobility shift assay (EMSA) showed that PDGF-BB stimulated protein binding to this motif that was inhibited in the presence of anti-Pea3 antibody. Analysis of these data provides the first evidence that PDGF-BB is a mitogen for MEOE-3M and increases CSF-1 protein levels, predominantly by transcription. Elucidation of the cellular pathways that control CSF-1 expression may provide novel strategies for the regulation of enamel matrix formation.

Caudal autotomy as anti-predatory behaviour in Palaeozoic reptiles.

Many lizards can drop a portion of their tail in response to an attack by a predator, a behaviour known as caudal autotomy. The capacity for intravertebral autotomy among modern reptiles suggests that it evolved in the lepidosaur branch of reptilian evolution, because no such vertebral features are known in turtles or crocodilians. Here we present the first detailed evidence of the oldest known case of caudal autotomy, found only among members of the Early Permian captorhinids, a group of ancient reptiles that diversified extensively and gained a near global distribution before the end-Permian  mass extinction event of the Palaeozoic. Histological and SEM evidence show that these early reptiles were the first amniotes that could autotomize their tails, likely as an anti-predatory behaviour. As in modern iguanid lizards, smaller captorhinids were able to drop their tails as juveniles, presumably as a mechanism to evade a predator, whereas larger individuals may have gradually lost this ability. Caudal autotomy in captorhinid reptiles highlights the antiquity of this anti-predator behaviour in a small member of a terrestrial community composed predominantly of larger amphibian and synapsid predators.

Predicting survival in patients with hepatocellular carcinoma: a UK perspective.

BACKGROUND AND AIMS: Hepatocellular carcinoma (HCC) is a cancer of rising incidence in the UK. The aim of this study was to compare the Okuda, Cancer of the Liver Italian Program (CLIP), and Barcelona Clinic Liver Cancer (BCLC) classifications as predictors of survival in UK patients with HCC. METHODS: Data were analysed from a prospective database maintained in a specialist hepatobiliary unit from 1998 to 2003. Each system was assessed for its discriminatory power, monotonicity of gradient, and independent contribution to prediction of mortality status based on a multivariate model. RESULTS: One hundred and two patients (77 males, 25 females) were identified with a median age of 65 (range, 14-87) years. The overall median survival time was 13 months and the one- and five-year survival rates were 52.9% (95% CI: 43.2%, 62.6%) and 35.3% (95% CI: 26.0%, 44.6%), respectively. All three classification systems had the capacity to differentiate between patient survival times across different stages. The Okuda system was superior in overall discriminatory power and in strength of monotonicity. The BCLC system, however, made the highest independent contribution of all three systems in predicting survival in the Cox regression model. CONCLUSIONS: All three classification systems were effective in predicting survival for patients with HCC in a UK population.

MEPE expression in osteocytes during orthodontic tooth movement.

UNLABELLED: MEPE and DMP1 may play a role in mineralisation and demineralisation within the osteocyte microenvironment. Our earlier studies showed that DMP1 is mechanically responsive [Gluhak-Heinrich J, Ye L, Bonewald LF, Feng JQ, MacDougall M, Harris SE, et al. Mechanical loading stimulates dentin matrix protein 1 (DMP1) in osteocytes in vivo. J Bone Min Res 2003;18(5):807-17]. OBJECTIVES: To examine the effect of mechanical loading on the expression of MEPE using mouse tooth movement model, and compare this effect to that on DMP1. METHODS: In situ hybridisation and immunohistochemistry was performed on 38 treated and 38 control bone sites loaded 6-72 h. ImageJ was used for quantification of mRNA expression in osteocytes. RESULTS: Alveolar osteocytes showed high basal level of MEPE that decreased during the first day of loading, followed by 2.8-fold stimulation at day 3, and returning to a control level by day 7. CONCLUSION: The osteocyte specific mechanical stimulation of MEPE was delayed and different, compared to that of DMP1. This suggests a distinct role of MEPE and DMP1 in the response of osteocytes to mechanical loading in vivo.

Targeted expression of csCSF-1 in op/op mice ameliorates tooth defects.

OBJECTIVE: The aim of this study was to characterize the tooth phenotype of CSF-1-deficient op/op mice and determine whether expression of csCSF-1 in these mice has a role in primary tooth matrix formation. DESIGN: Ameloblasts and odontoblasts, isolated from wt/wt frozen sections using laser capture microdissection, were analysed for csCSF-1, sCSF-1 and CSF-1R mRNA by RT-PCR. Mandibles, excised from 8 days op/op and wt/wt littermates, were examined for tooth morphology as well as amelogenin and DMP1 expression using in situ hybridisation. op/opCS transgenic mice, expressing csCSF-1 in teeth and bone using the osteocalcin promoter, were generated. Skeletal X-rays and histomorphometry were performed; teeth were analysed for morphology and matrix proteins. RESULTS: Normal dental cells in vivo express both CSF-1 isoforms and CSF-1R. Compared to wt/wt, op/op teeth prior to eruption showed altered dental cell morphology and dramatic reduction in DMP1 transcripts. op/opCS mice showed marked resolution of osteopetrosis, tooth eruption and teeth that resembled amelogenesis imperfecta-like phenotype. At 3 weeks, op/op teeth showed severe enamel and dentin defects and barely detectable amelogenin and DMP1. In op/opCS mice, DMP1 in odontoblasts increased to near normal and dentin morphology was restored; amelogenin also increased. Enamel integrity improved in op/opCS, although it was thinner than wt enamel. CONCLUSIONS: Results demonstrate that ameloblasts and odontoblasts are a source and potential target of CSF-1 isoforms in vivo. Expression of csCSF-1 within the tooth microenvironment is essential for normal tooth morphogenesis and may provide a mechanism for coordinating the process of tooth eruption with endogenous matrix formation.

A survey regarding acceptability of oral emergency contraception according to the posited mechanism of action.

OBJECTIVE: The objective was to determine the acceptability to women of oral emergency contraception (EC) that works by inhibiting ovulation, preventing implantation or disrupting implantation, and also to determine the characteristics of women associated with the acceptability of each posited mechanism of action. STUDY DESIGN: Women completed a self-administered, anonymous questionnaire asking whether they would consider using an EC pill based on each of three hypothetical mechanisms of action: inhibiting ovulation, preventing implantation or disrupting implantation. The questionnaire was distributed among women in Edinburgh, UK, (a) presenting for EC at a community pharmacy, (b) attending a clinic for insertion of intrauterine contraception (IUC) or (c) attending a clinic for an induced abortion. Descriptive analyses stratified women according to healthcare setting and personal characteristics. Univariable and multivariable analyses were used to establish factors which may predict acceptability of each EC pill's mechanism of action. RESULTS: Four hundred and nineteen out of 458 (91%) women responded to the survey. Overall, women reported that EC would be acceptable if it worked by inhibiting ovulation (89%), preventing implantation (83%) or disrupting implantation (75%). Among women seeking abortion, more would accept an EC pill which disrupted implantation compared to women seeking IUC (odds ratio, 2.19; 95% confidence interval, 1.30-3.69; p=.004). Based on multivariable analyses, factors associated with acceptability included previous use of EC, previously holding strong views against abortion and having had a previous abortion. CONCLUSION: For each of the posited mechanisms of action, a majority of women surveyed would be willing to consider oral EC to prevent unintended pregnancy. IMPLICATIONS STATEMENT: The scope of the study was limited, and further work on the views of women in the wider population is needed. This is important as the development of such drugs to prevent pregnancy is likely to raise political and ethical challenges, particularly in relation to disruption of implantation.

Clinical features remain important for the diagnosis of infective endocarditis in the modern era.

BACKGROUND: Infective endocarditis (IE) can be difficult to diagnose, due to multiple (often non-specific) presenting features. AIM: To assess the predictive accuracy of classical clinical features and blood investigations readily available at the time of presentation. DESIGN: Cross-sectional analysis. METHODS: We studied 29 IE cases and 79 controls (clinically suspicious contemporaneous cases where IE was subsequently excluded) from a hospital-based group of patients referred to a cardiac department with possible infective endocarditis. Patients were identified from the echocardiography database. Cases were defined by final diagnosis. Symptoms, signs, risk factors for IE and blood investigations were recorded from case notes and examined by univariate and multivariate analyses. RESULTS: The sensitivity, specificity, and positive and negative predictive values of transthoracic echocardiography (TTE) for detection of IE in clinically suspected cases were 71%, 98%, 57% and 99%, respectively. Univariate analyses revealed a significant association between IE and several clinical features. Under multivariate analysis, previous heart valve surgery (OR 13.3, 90%CI 3.2-55.6), positive blood cultures (OR 17.2, 90%CI 4.9-58.8), signs of embolism (OR 11.4, 90%CI 3.0-43.5), a new, altered or changing murmur (OR 10.3, 90%CI 2.8-38.5) and splenomegaly (OR 18.2, 90%CI 3.6-90.9) were independent predictors for IE. DISCUSSION: Clinical features at presentation continue to be important for the diagnosis of IE. Features such as positive blood cultures, signs of embolism and a changing heart murmur should be used to guide investigation and treatment of IE prior to echocardiography, or when TTE is negative.

G1 expression and multistage dynamics of cyclin A in human osteosarcoma cells.

Cyclin A was initially characterized as a 'mitotic cyclin', believed to function exclusively at the G2-to-M phase transition; however, recent studies have provided compelling evidence that cyclin A additionally functions earlier in the mammalian somatic cell cycle as a putative 'S-phase-promoting factor'. Moreover, numerous inconsistencies have arisen concerning the temporal induction, subcellular localization, subunit configuration, covalent modification and proteolytic destruction of cyclin A, as well as the physiological function of the cyclin A-associated protein kinase complexes. Utilizing precisely synchronized human MG-63 osteosarcoma cells, the present study demonstrates that cyclin A mRNA and protein are clearly expressed in late G1 prior to S-phase entry, as is cyclin A-associated kinase activity and concomitant phosphorylation of the Rb protein. A series of monospecific cyclin A antibodies were generated and utilized to confirm that multiple covalent modifications of cyclin A occur during the course of the cell cycle, and to characterize the subcellular dynamics in additional detail. Pharmacological blockade with mimosine was utilized to further delineate cyclin A expression and to distinguish the temporal induction from the mechanisms of enzyme activation. Subcellular fractionation and immunocytochemical staining localized nascent cyclin A to the cytoplasm, and revealed a distinct translocation to the nucleus during the G1-to-S phase transition. The results of these studies support a multistage model of cyclin A metabolism and enzyme activation.

Effect of angiotensin-converting enzyme inhibition on renal function and albuminuria in normotensive type I diabetic patients.

Normotensive patients with insulin-dependent (type I) diabetes mellitus (n = 18) were given 25 mg captopril (b.i.d.) and placebo for 3 mo in a randomized double-blind crossover study. Patients had normal renal function, and none had retinopathy. Albuminuria was less than 20 micrograms/min in 12 patients and between 20 and 200 micrograms/min in the other 6. Patients were examined at the end of the placebo and captopril phases. Captopril caused little reduction in blood pressure obtained by 24-h ambulatory monitoring (systolic 126.0 +/- 2.7 to 123.9 +/- 2.4 mmHg, P less than 0.08; diastolic 74.2 +/- 1.9 to 72.1 +/- 1.9 mmHg, P less than 0.09). Captopril lowered glomerular filtration rate from 99.5 +/- 7.7 to 71.0 +/- 5.5 ml.min-1. 1.73 m-2 (P less than 0.01), whereas renal plasma flow (443.9 +/- 15.2 ml.min-1. 1.73 m-2) remained unchanged. Filtration fraction was reduced from 22.4 +/- 1.4 to 17.4 +/- 1.4% (P less than 0.01). Urinary albumin excretion was reduced from 59.1 +/- 0.15 to 27.7 +/- 13.9 micrograms/min (P less than 0.1). Reduction was related to the extent of initial albuminuria (r = 0.997, P less than 0.001), a relationship that remained significant after logarithmic transformation (r = 0.540, P less than 0.02). Dextran clearance was used to determine glomerular capillary function. Angiotensin inhibition caused reduction in effective glomerular pore size and also reduced flow via the nondiscriminatory shunt. Angiotensin inhibition in normotensive patients with type I diabetes was well tolerated. Reduction in albuminuria is mediated by a combination of hemodynamic changes and alterations in glomerular capillary function.

Recognition of hypertension and abnormal blood pressure burden with ambulatory blood pressure recordings in type I diabetes mellitus.

Ambulatory blood pressure (AMBP) measurements were obtained at 20-min intervals for 24 h in 25 subjects with insulin-dependent (type I) diabetes mellitus and 21 control subjects. The diabetic patients had normal kidney function (glomerular filtration rate 112.1 +/- 7.2 ml.min-1.1.73 m-2, renal plasma flow 459.0 +/- 23.4 ml.min-1.1.73 m-2) and were normotensive according to standard sphygmomanometer examinations. Mean +/- SE AMBP (systolic/diastolic in mmHg) measurements in diabetic patients (24 h, 131.7/77.2 +/- 2.9/1.8; 0600-2200, 132.3/78.4 +/- 2.9/3.4; 2200-0600, 125.1/75.7 +/- 3.9/3.4) significantly exceeded control values during all times (24 h, 121.8/70.3 +/- 2.9/1.9; 0600-2200, 120.7/71.8 +/- 2.6/2.0; 2200-0600, 108.2/61.5 +/- 6.6/2.7). Mean 24-h AMBP exceeded 135/85 mmHg in 49% of diabetic patients. The same threshold of 135/85 mmHg was used to determine the prevalence of abnormal measurements per time period (pressure burden). Pressure burden was increased twofold in diabetic patients compared with control subjects. Mean AMBP was significantly reduced at night in control subjects but not in diabetic patients. Changes in blood pressure were not related to kidney function in diabetic patients. AMBP recordings uncovered an increased prevalence of abnormal mean blood pressure, increased pressure burden, and a lack of diurnal variation of blood pressure in subjects with type I diabetes mellitus. These findings have important implications for early intervention strategies in diabetes mellitus because AMBP recordings correlate well with end-organ damage.