Impact of genetic polymorphisms on the smoking-related risk of periodontal disease: the population-based study SHIP.

Periodontitis is a bacterial inflammatory disease leading to attachment loss with the consequence of tooth loss. There exists a multifactorial risk pattern including bacterial challenge, smoking, age, sex, diabetes, socio-economic and genetic factors. Smoking has the highest impact on the course of the disease modulated by all the other factors. Here, we report the relationship between smoking and the polymorphisms of genetic polymorphisms inflicted in the pathogenesis.In a randomly selected population-based study, 1083 subjects were typed for the polymorphisms of the IL-1 genotype, Fcgamma RIIIb receptor gene, myeloperoxidase and N-acetyltransferase (NAT2) and related to their periodontal state. Smoking behavior was assessed including present and past quality and quantity of smoking.There is a significant dose-effect relationship between the exposure to tobacco smoke and the extent of periodontal disease assessed as attachment loss and tooth loss. Moreover, there are gene-environmental interactions as subjects bearing variant genotypes show an enhanced smoking-associated risk of the disease modulated by these genotypes. In non-smokers, the impact of these genetic polymorphisms is mostly negligible.This study provides support for the hypothesis that subjects bearing genetic variants of polymorphically expressed phenotypes are at an increased risk of periodontitis when smoking. Mostly, this may be accomplished via the influence of smoking-related impairment on defense mechanisms rather than on the pathogenic pathways.

Nonmuscle myosin heavy chain IIA mutations define a spectrum of autosomal dominant macrothrombocytopenias: May-Hegglin anomaly and Fechtner, Sebastian, Epstein, and Alport-like syndromes.

May-Hegglin anomaly (MHA) and Fechtner (FTNS) and Sebastian (SBS) syndromes are autosomal dominant platelet disorders that share macrothrombocytopenia and characteristic leukocyte inclusions. FTNS has the additional clinical features of nephritis, deafness, and cataracts. Previously, mutations in the nonmuscle myosin heavy chain 9 gene (MYH9), which encodes nonmuscle myosin heavy chain IIA (MYHIIA), were identified in all three disorders. The spectrum of mutations and the genotype-phenotype and structure-function relationships in a large cohort of affected individuals (n=27) has now been examined. Moreover, it is demonstrated that MYH9 mutations also result in two other FTNS-like macrothrombocytopenia syndromes: Epstein syndrome (EPS) and Alport syndrome with macrothrombocytopenia (APSM). In all five disorders, MYH9 mutations were identified in 20/27 (74%) affected individuals. Four mutations, R702C, D1424N, E1841K, and R1933X, were most frequent. R702C and R702H mutations were only associated with FTNS, EPS, or APSM, thus defining a region of MYHIIA critical in the combined pathogenesis of macrothrombocytopenia, nephritis, and deafness. The E1841K, D1424N, and R1933X coiled-coil domain mutations were common to both MHA and FTNS. Haplotype analysis using three novel microsatellite markers revealed that three E1841K carriers--one with MHA and two with FTNS--shared a common haplotype around the MYH9 gene, suggesting a common ancestor. The two new globular-head mutations, K371N and R702H, as well as the recently identified MYH9 mutation, R705H, which results in DFNA17, were modeled on the basis of X-ray crystallographic data. Altogether, our data suggest that MHA, SBS, FTNS, EPS, and APSM comprise a phenotypic spectrum of disorders, all caused by MYH9 mutations. On the basis of our genetic analyses, the name "MYHIIA syndrome" is proposed to encompass all of these disorders.

Polymorphisms of Fc gamma-receptors RIIa, RIIIa, and RIIIb in patients with adult periodontal diseases.

Polymorphisms influencing the binding affinity between the Fcgamma receptors and IgG of different subclasses are thought to be of importance in the individual susceptibility to infections with Gram-negative bacteria contributing to periodontal disease. One hundred and fifty-four Caucasian subjects were clinically and radiographically examined for their periodontal status and genotyped for their allelic pattern of FcgammaRIIa, FcgammaRIIIa, and FcgammaIIIb polymorphism. In assessing periodontitis according to mean probing depth and attachment loss, no differences were found in allele frequencies or combined allotypes between the subjects with mild or moderate and those with severe signs of periodontitis. However, the extent and severity of bone loss were significantly associated with the genotype of the receptor FcgammaRIIIa. An increased risk of severe bone destruction was observed in individuals carrying the FcgammaRIIIa-VV genotype (OR = 5.3; 95% CI 1.4-26.2). FcgammaRIIIb is in linkage disequilibrium with FcgammaRIIIa. Hence it is also related to periodontal disease. There is no indication of an association between the polymorphism of FcgammaRIIa and periodontitis. The results are evidence that the FcgammaRIIIa genotype coding for the high affinity receptor imposes an additional risk of bone loss as does the FcgammaRIIIb genotype coding for the low affinity receptor.

Very low platelet counts in post-transfusion purpura falsely diagnosed as heparin-induced thrombocytopenia. Report of four cases and review of literature.

Differential diagnosis between post-transfusion purpura (PTP) and heparin-induced thrombocytopenia (HIT) can be difficult in the initial stages of thrombocytopenia, as the early clinical presentations are often similar. Four patients are described who were suspected clinically of suffering from HIT. All four patients had recent blood transfusions and platelet alloantibodies, thus the diagnosis of PTP was made. One lethal gastrointestinal and one retroperitoneal hemorrhage developed in two of the four patients. Unusually, one patient was male and two different platelet alloantibodies were present in his serum; in another patient platelet alloantibodies and HIT-antibodies were detectable. To arrive at the right diagnosis as quickly as possible is vitally important since treatment, which has to be initiated promptly, is very different for the two syndromes. Thus, we suggest that in patients where HIT is suspected, additional information should be sought. If features consistent with PTP (such as a recent blood transfusion or a marked drop in platelet count to below 15 Gpt/L) are present, we recommend parallel testing for platelet alloantibodies to rule out PTP.

Autosomal-dominant giant platelet syndromes: a hint of the same genetic defect as in Fechtner syndrome owing to a similar genetic linkage to chromosome 22q11-13.

Families with 3 different syndromes characterized by autosomal dominant inheritance of low platelet count and giant platelets were studied. Fechtner syndrome is an autosomal-dominant variant of Alport syndrome manifested by nephritis, sensorineural hearing loss, and cataract formation in addition to macrothrombocytopenia and polymorphonuclear inclusion bodies. Sebastian platelet syndrome is an autosomal-dominant macrothrombocytopenia combined with neutrophil inclusions that differ from those found in May-Hegglin syndrome or Chediak-Higashi syndrome or the Dohle bodies described in patients with sepsis. These inclusions are, however, similar to those described in Fechtner syndrome. Other features of Alport syndrome, though, including deafness, cataracts, and nephritis, are absent in Sebastian platelet syndrome. Epstein syndrome is characterized by macrothrombocytopenia without neutrophil inclusions, in addition to the classical Alport manifestations-deafness, cataracts, and nephritis-and it is also inherited in an autosomal-dominant mode. We mapped the disease-causing gene to the long arm of chromosome 22 in an Italian family with Fechtner syndrome, 2 German families with the Sebastian platelet syndrome, and an American family with the Epstein syndrome. Four markers on chromosome 22q yielded an LOD score greater than 2.76. A maximal 2-point LOD score of 3.41 was obtained with the marker D22S683 at a recombination fraction of 0.00. Recombination analysis placed the disease-causing gene in a 3.37-Mb interval between the markers D22S284 and D22S693. The disease-causing gene interval in these 3 syndromes is similar to the interval described recently in an Israeli family with a slightly different Fechtner syndrome than the one described here. Recombination analysis of these 3 syndromes refines the interval containing the disease-causing gene from 5.5 Mb to 3.37 Mb. The clinical likeness and the similar interval containing the disease-causing gene suggest that the 3 different syndromes may arise from a similar genetic defect.

M-like proteins of Streptococcus dysgalactiae.

Streptococcus dysgalactiae is one of the most important bacterial species isolated from bovine mastitis. To identify potential virulence factors of this species we prepared chromosomal DNA from strain 8215 and constructed a phage display library. By affinity selection of the library against fibrinogen (Fg), we isolated and characterized a gene, called demA, encoding a protein with the molecular mass of approximately 58 kDa, called DemA, displaying both plasma protein binding properties and sequence similarities with the M and M-like proteins of other streptococcal species. Purified recombinant DemA protein was found to completely inhibit Fg-binding to cells of S. dysgalactiae. A continued sequence analysis revealed that the demA gene was preceded by an open reading frame (dmgA) coding for a putative protein, called DmgA, with high similarities to the Mga proteins of Streptococcus pyogenes. By additional cloning, the corresponding dmgA and demA genes from another strain, called Epi9, were isolated and analyzed. These genes, called dmgB and demB, respectively, revealed a high degree of similarity to the corresponding genes in strain 8215. Increased binding of Fg by cells of strain Epi9, grown in an atmosphere with 10% CO(2), was correlated to an enhanced transcription of the demB gene as shown in a Northern blot. Strain 8215 did not respond to CO(2), which could be explained by a nonfunctional dmgA gene due to insertion of an insertion sequence element. Based on sequence similarities of the described proteins to Mga, M, and M-like proteins and the response to elevated level of CO(2), we suggest that the dmg and dem genes are members of a regulon similar to the described mga regulon in S. pyogenes, which encodes several virulence factors in this species.

The alpha2 gene coding sequence T807/A873 of the platelet collagen receptor integrin alpha2beta1 might be a genetic risk factor for the development of stroke in younger patients.

The polymorphisms C807T and G873A of the platelet integrin alpha2beta1 (collagen receptor glycoprotein [GP] Ia-IIa) are linked to the expression density of this receptor. The GPIa T807/A873 allele causes a higher receptor expression, enhancing platelet binding to collagen. This might present a genetic predisposition for the development of thromboembolic complications. In this case-control study, the genotypes of the GPIa C807T polymorphism and presence of conventional risk factors (hypertension, diabetes mellitus, and smoking) were compared in stroke patients and patients without cerebrovascular disease (non-CVD patients)

Heparin-induced thrombocytopenia: new insights into the impact of the FcgammaRIIa-R-H131 polymorphism.

Heparin-induced thrombocytopenia (HIT), a severe complication of heparin treatment, can be associated with new thrombotic complications. HIT antibodies activate platelets via the platelet Fcgamma-receptor (FcgammaRIIa), which carries a functionally relevant polymorphism (FcgammaRIIa-R-H131). The effect of this polymorphism on the clinical manifestations of HIT is controversial. We determined prospectively the FcgammaRIIa-R-H131 genotypes in 389 HIT patients, in 351 patients with thrombocytopenia or thrombosis due to causes other than HIT and without detectable HIT antibodies, and in 256 healthy blood donors. For this purpose, a novel nested sequence-specific primer-polymerase chain reaction (SSP-PCR) was developed. FcgammaRIIa-R/R131 was found to be overrepresented in the HIT patients (27%) compared with the control groups (non-HIT patients [21%] and blood donors [20%]). In a subgroup of 122 well-characterized HIT patients, the genotype distribution in patients presenting with thrombocytopenia only was compared with that of patients who developed thromboembolic complications. The frequency of FcgammaRIIa-R/R131 among patients with thrombotic events was significantly elevated (37% v 17%; P = .036). Our results indicate that genotype distribution can be correlated to the clinical outcome of patients with HIT. We speculate that the reduced clearance of immune complexes in patients with the FcgammaRIIa-R/R131 allotype causes prolonged activation of endothelial cells and platelets, thus increasing the risk for thrombotic complications.

Polymorphisms of the human platelet antigens HPA-1, HPA-2, HPA-3, and HPA-5 on the platelet receptors for fibrinogen (GPIIb/IIIa), von Willebrand factor (GPIb/IX), and collagen (GPIa/IIa) are not correlated with an increased risk for stroke.

BACKGROUND AND PURPOSE: A recent study has described a high incidence of the human platelet antigen (HPA)-1b alloantigen in patients with myocardial infarction. We investigated the distribution of gene polymorphisms of platelet glycoproteins (GPs) in patients with cerebrovascular disease (CVD) and stroke. The polymorphic systems we have studied are HPA-1 and HPA-3 on the fibrinogen receptor (GPIIb/IIIa), HPA-2 on the von Willebrand factor receptor (GPIb/IX), and HPA-5 on one of the platelet collagen receptors (GPIa/IIa). METHODS: DNA was isolated from peripheral blood collected from 218 consecutive stroke patients, 165 neurological inpatients without signs of CVD, and 321 healthy blood donors. The genotypes of HPA-1, HPA-2, HPA-3, and HPA-5 were determined by sequence specific primer polymerase chain reactions. RESULTS: The calculated allele frequencies were as follows: for CVD patients, HPA-1a/b 0.81/0.19, HPA-2a/b 0.91/0.09, HPA-3a/b 0.61/0.39, and HPA-5a/b 0.92/0.08; for inpatient HPA-1a/b 0.83/0.17, HPA-2a/b 0.91/0.09, HPA-3a/b 0.62/0.3 and HPA-5a/b 0.93/0.07; and for blood donors, HPA-1a 0.85/0.15, HPA-2a/b 0.94/0.06, HPA-3a/b 0.60/0.40, and HPA 5a/b 0.92/0.08. There were no statistically significant difference for the analyzed HPA polymorphism frequencies either between the CVD patients and the non-CVD inpatients or the CVD patients and blood donors. However, the HPA-1b genotype was slightly more frequent in patients (CVD and non-CVD) than in the healthy blood donors. CONCLUSIONS: Our results indicate that the HPA-1, HPA-2, HPA-3, and HPA-5 polymorphisms are not associated with an increased risk for stroke.

The tobacco homolog of mammalian calreticulin is present in protein complexes in vivo.

The analysis of protein sorting signals responsible for the retention of reticuloplasmins (RPLs), a group of soluble proteins that reside in the lumen of the endoplasmic reticulum (ER), has revealed a structural similarity between mammalian and plant ER retention signals. We present evidence that the corresponding epitope is conserved in a vast family of soluble ER resident proteins. Microsequences of RPL60 and RPL90, two abundant members of this family, show high sequence similarity with mammalian calreticulin and endoplasmin. RPL60/calreticulin cofractionates and costains with the lumenal binding protein (BiP). Both proteins were detected in the nuclear envelope and the ER, and in mitotic cells in association with the spindle apparatus and the phragmoplast. Immunoprecipitation of proteins from in vivo-labeled cells demonstrated that RPL60/calreticulin is associated with other polypeptides in a stress- and ATP-dependent fashion. RPL60/calreticulin transcript levels increased rapidly in abundance during the proliferation of the secretory apparatus and the onset of hydrolase secretion in gibberellic acid-treated barley aleurone cells. This induction profile is identical to that of the well-characterized ER chaperones BiP and endoplasmin. However, expression patterns in response to different stress conditions as well as tissue-specific expression patterns indicate that these genes are differentially regulated and may not act in concert.

Clinical findings related to morphologic changes in TMJ autopsy specimens.

Numerous temporomandibular joint autopsy studies have been presented in the literature for the last two decades, but signs and symptoms of temporomandibular disorders before death were not available. To investigate the clinical significance of morphologic changes in the temporomandibular joint, 19 persons were clinically examined for signs and symptoms of temporomandibular disorders. The temporomandibular joints were subsequently analyzed macroscopically at autopsy and statistically associated with history and clinical findings. The average time between clinical examination and autopsy was 12 months. Signs and symptoms of temporomandibular disorders were not common findings for these persons. Morphologically, 31 of the 34 joints showed different forms of changes such as deviation in form, arthrosis, disk displacement, disk deformation, and adhesions. Crepitation showed a significant association with arthrosis. It was concluded that the association between pain and dysfunction and joint morphology is complex and gross morphologic alterations can be present in the absence of temporomandibular joint pain and dysfunction.

Estimation of the possibility to treat the edentulous maxilla with osseointegrated implants.

The possibility to treat individuals with an edentulous maxilla with osseointegrated implants was studied in randomly selected individuals from the city of Malmö. From a panoramic radiograph one oral radiologist and one oral surgeon together decided possible number and length of implants potentially installable. It was found that treatment with osseointegrated implants could possibly be done in 80% of the individuals and in 20% bone transplantation would have been necessary. Preoperative tomography was needed in about two thirds of the individuals for a more accurate evaluation of the degree of bone resorption.

Histologic features of the temporomandibular joint disk and posterior disk attachment: comparison of symptom-free persons with normally positioned disks and patients with internal derangement.

To gain further knowledge about the differences between normal and pathologic anatomy of the temporomandibular joint disk, we examined histologically disks obtained at autopsy from 10 symptom-free persons and compared our findings with observations involving 17 surgically removed disks. The surgical patients had internal derangement and severe long-standing temporomandibular joint pain and dysfunction. The normal disks were biconcave, whereas the surgically removed disks were deformed and thicker than the normal disks. Chondrocytes (4 joints), a surface layer of proliferative connective tissue (4 joints), vessels (2 joints), and splitting (4 joints) were seen in the surgical specimens but not in the normal specimens. The surgical specimens also showed higher maximal density of fibroblasts and vessels. It was concluded that surgically removed temporomandibular joint disks demonstrated several histologic alterations that were not seen in normal disks. These characteristics may serve as the basis for development of a histologic grading of pathologic conditions of the temporomandibular joint disk and the posterior disk attachment.

Quantitative correlative proton and electron microprobe analysis of biological specimens.

To investigate the possibility of quantitative correlative proton microprobe (PMP) and electron microprobe (EMP) analysis of biological soft tissue, model specimens were analyzed by both techniques. The specimens consisted of freeze-dried sections of gelatin containing known concentrations of nickel chloride. Both for PMP and for EMP, the signal was expressed as the ratio of the characteristic intensity and the continuum intensity in a peak-free region of the spectrum. With both techniques, calibration curves (signal versus known concentration) obtained, showed a deviation from linearity at high nickel concentrations. However, a linear relation (correlation coefficient 0.996) was obtained in a plot of EMP signal versus PMP signal. This indicates that quantitative correlative PMP and EMP analysis can be carried out by using the same standard for both analytical techniques.

Elemental analysis on freeze-dried sections of human skin: studies by electron microprobe and particle induced X-ray emission analysis.

The possibility of using the proton microprobe (PMP) in the elemental analysis of dermatological material, under conditions where the spatial resolution can be restricted to a few micrometers, was demonstrated. Comparison with the electron microprobe (EMP) technique using duplicate sections from the same human skin biopsies revealed that the PMP and EMP techniques are complementary and yield closely corresponding elemental distributions for elements of dermatological interest. The concentrations of phosphorus and potassium were low in the dermis, high in the stratum basale and stratum spinosum, and decreased markedly in the stratum granulosum to a low level in the stratum corneum. The sulfur concentration was highest in the stratum corneum.

Particle-induced X-ray emission (PIXE) analysis of trace elements in human coronal dentin.

Mass concentrations reported on the elemental composition of human dentin show great discrepancies between different investigations using various techniques. The purpose of the study was to determine the concentrations of bromine (Br), copper (Cu), iron (Fe), strontium (Sr) and zinc (Zn) in the coronal dentin of permanent teeth from Sweden and from New York City. Particle-induced X-ray emission (PIXE) was used for the analyses. There were no statistically significant differences in elemental composition between the Swedish and the New York teeth, although the ranges were wider in the American material, except for Br. The results were compared with values presented by other investigators. The significance of the differences noted is uncertain in view of the differences in materials and methods employed in earlier studies. The PIXE technique applied to the analysis of trace elements in dental hard tissues offers great advantages in being accurate, rapid, and nondestructive.

Proton-induced X-ray emission analysis - a new tool in quantitative dermatology.

Proton-Induced X-ray Emission analysis (PIXE) constitutes a method for trace element analysis characterized by multielemental capability, detection limits in the low ppm-range and size resolution down towards a micrometre. In applications where the sensitivity of the Electron-Induced X-ray Emission (EIXE) analysis is not sufficient and where a spatial resolution not better than a few micrometres is required, the PIXE technique provides a powerful tool. In this paper properties of the PIXE method are demonstrated by quantitative results from three different samples of dermatological interest. Firstly, mercury results from a longitudinal scan of a single hair strand from a mercury poisoned person are shown. With a spatial resolution of one or a few millimetres very fast scans may be performed on hair strands giving information on time and magnitude of intoxication or other exposures, as well as deficiencies. Secondly, results are given from a radial scan with a beam width of 4 micron on hair from a person exposed to high amounts of iron. The calcium, iron and zinc distributions but not the sulphur and potassium distributions show narrow peaks of concentration (less than 4 micron) about 15 micron from the surface of the hair. Further investigations have to be performed in order to interpret these data. Thirdly, the depth profiles in skin of some elements were measured with a beam width of 10 micron. The results show significant increases in sulphur, calcium and zinc concentrations and significant decreases in phosphorous and potassium concentrations at the skin surface, i.e. in the stratum corneum.

Lead levels determined in Swedish permanent teeth by particle-induced X-ray emission.

The determination of lead in permanent teeth is a useful measure of past exposure in early childhood since these teeth are mineralized in early childhood. Particle-induced X-ray emission (PIXE) analysis has been shown to be a method with good applicability for the contamination-free analysis of elements heavier than calcium in dental hard tissues. The method is rapid and nondestructive. The purpose of this study, which used the PIXE technique, was to survey the average level of lead in the coronal dentin of permanent bicuspid teeth collected in three places representing Swedish urban and rural areas. In addition teeth from the New York City area were analyzed. The material comprised 165 teeth from Sweden and, for comparison, 14 teeth from New York City. The median value of lead in the Swedish teeth was 2.9 micrograms/g, a value indicating an insignificant influence from the environment in comparison to the New York teeth, for which the median value was 9.2 micrograms/g. There was however a statistically significant difference in the lead concentration of teeth from large and small Swedish cities; this finding may reflect different automobile traffic intensity.

Anatomy of mandibular third molars. A comparison between radiographic appearance and clinical observations.

The purpose of this study was to present a radiographic technique for the examination of mandibular third molars and to investigate the extent of correspondence between the radiographic appearance and the true anatomy of the tooth. Forty-four mandibular third molars were examined radiographically and subsequently removed. The investigation showed that radiographic examination of mandibular third molars, utilizing intraoral films in three different projections, gave an insight into the true anatomy of the tooth. The few instances of minor misinterpretation could be grouped into two categories: (1) root dilacerations which paralleled the direction of the beam were not always apparent on the radiographs; and (2) the degree of fusion between roots lying close together was not usually well defined on the radiographs.