The state of the oral health in patients with type 2 diabetes.

Over 3 mln people in Poland suffer from diabetes out of which 1/3 are undiagnosed. Diabetes is a metabolic disease characterized by the chronic hyperglycaemia resulting from disturbances in insulin secretion and tissue resistance to its action. The most serious clinical problem in the diabetics is periodontal disease since hyperglycaemia influences the disturbance of the immune response and increases the number of periopathogens. AIM: The aim of the study was an assessment of the oral health in patients with type 2 diabetes. MATERIALS AND METHODS: The study was conducted in the group of 450 people who reported to the selected specialist medical centers at the Rural Medicine Institute. Adult patients from Lublin macroregion were referred to the Department. 153 people among the studied were Diabetic Clinic patients with type 2 diabetes The assessment of the oral health was carried out on the basis of clinical examination included DMFt - the number of teeth with decay (D), the number of teeth removed because of tooth decay (M) or filled (F), index and masticatory function loss index (t). The results are summarized in tables and subjected to statistical analysis. A non- parametric Kruskall-Wallis test was used. RESULTS: In the group up to 34 years of age the number of extracted teeth was higher in healthy people. The index of masticatory function loss below 100% was higher in the diabetic patients. CONCLUSIONS: Diabetic outpatient clinic patients should receive special dental care and frequent check-ups.

Periodontal condition in patients of the specialist Outpatient Clinics at the Institute of Rural Health in Lublin, Poland.

INTRODUCTION: Periodontal disease is a chronic inflammation which, if remains untreated, can lead to the loss of teeth and supporting structures. Evidence data support the relationship of periodontal disease with the development and course of diseases such as heart attack, stroke, hypertension, chronic renal diseases, rheumatoid arthritis or diabetes. OBJECTIVE: The aim of the study was to conduct an assessment of periodontal status and periodontal needs in people from the rural environment who were patients of selected specialist outpatient clinics at the Institute of Rural Health in Lublin, Poland. MATERIAL AND METHODS: The examined population included 450 patients. The Community Periodontal Index of Treatment Needs, which is a measure of the assessment of the selected periodontal symptoms incidence, was used. The obtained data was discussed and analyzed with Chi-square test. RESULTS: The data obtained revealed that a healthy periodontium occurred only in 5.1% of respondents, tartar in 41.6%, pathological pockets of 3.5-5.5 mm in 23.6%, and pockets deeper than 5.5 mm in 5.8% of patients. Most people with healthy periodontium were in the youngest age group. In the analyzed group, 7.1% of patients required specialized comprehensive periodontal treatment, and only 6.5% of the examined persons did not show any need for periodontal treatment. CONCLUSIONS: Patients of specialist clinics of the Institute of Rural Health who formed the analyzed group, had affected periodontium which required comprehensive periodontal treatment. The alarmingly high percentage of people over 55 years of age with advanced periodontopathy may translate into an increased risk of cause-and-effect incidence of systemic diseases.

Using the comet assay and lysis conditions to characterize DNA lesions from the acrylamide metabolite glycidamide.

The alkaline comet assay and a cell-free system were used to characterise DNA lesions induced by treatment with glycidamide (GA), a metabolite of the food contaminant acrylamide. DNA lesions induced by GA were sensitively detected when the formamidopyrimidine-DNA-glycosylase (Fpg) enzyme was included in the comet assay. We used LC-MS to characterise modified bases from GA-treated naked DNA with and without subsequent Fpg treatment. N7-GA-Guanine and N3-GA-Adenine aglycons were detected in the supernatant showing some depurination of adducted bases; treatment of naked DNA with Fpg revealed no further increase in the adduct yield nor occurrence of other adducted nucleobases. We treated human lymphocytes with GA and found large differences in DNA lesion levels detected with Fpg, depending on the duration and the pH of the lysis step. These lysis-dependent variations in GA-induced Fpg sensitive sites paralleled those observed after treatment of cells with methyl methane sulfonate (MMS). On the other hand, oxidative lesions (8-oxoGuanine) induced by a photoactive compound (Ro 12-9786) plus light, and also DNA strand breaks induced by X-rays, were detected largely independently of the lysis conditions. The results suggest that the GA-induced lesions are predominantly N7-GA-dG adducts slowly undergoing imidazole ring opening at pH 10 as in the standard lysis procedure; such structures are substrate for Fpg leading to strand breaks. The data suggest that the characteristic alkaline lysis dependence of some DNA lesions may be used to study specific types of DNA modifications. The comet assay is increasingly used in regulatory testing of chemicals; in this context, lysis-dependent variations represent a novel approach to obtain insight in the molecular nature of a genotoxic insult.

Identification of adducts formed in the reactions of malonaldehyde-glyoxal and malonaldehyde-methylglyoxal with adenosine and calf thymus DNA.

The reactions of adenosine with malonaldehyde and glyoxal, and with malonaldehyde and methylglyoxal resulted in the formation of one malonaldehyde-glyoxal and one malonaldehyde-methylglyoxal conjugate adduct, respectively. These adducts were isolated and purified by reversed-phase liquid chromatography, and structurally characterized by UV, (1)H- and (13)C-NMR spectroscopy, and mass spectrometry. The malonaldehyde-glyoxal adduct was identified as 8-(diformylmethyl)-3-(beta-D-ribofuranosyl)imidazo[2,1-i]purine (M(1)Gx-A), while the malonaldehyde-methylglyoxal one as 8-(diformylmethyl)-7-methyl-3-(beta-D-ribofuranosyl)imidazo[2,1-i]purine (M(1)MGx-A). Both adducts were also observed in calf thymus DNA when incubated in the respective aldehydes under physiological pH and temperature. Moreover, in the reaction of methylglyoxal and malonaldehyde with adenosine, an additional adduct was formed. This adduct was found to consist of one unit derived from methylglyoxal and one unit from formaldehyde. The adduct was identified as N(6)-(2,3-dihydroxy-2-methylpropanoyl)-9-(beta-D-ribofuranosyl)purine (MGxFA-A). Formaldehyde was found to originate from the commercial methylglyoxal in which it was present as an impurity.

Reactions of 4-[Bis(2-chloroethyl)amino]benzenebutanoic acid (chlorambucil) with DNA.

4-[Bis(2-chloroethyl)amino]benzenebutanoic acid (=chlorambucil, 1; 2.5 mM) was allowed to react with single- and double-stranded calf thymus DNA at physiological pH (cacodylic acid, 50% base) at 37 degrees . The DNA-chlorambucil adducts were identified by analyzing the DNA hydrolysates by NMR, UV, HPLC, LC/ESI-MS/MS techniques as well as by spiking with authentic materials. ssDNA was more reactive than dsDNA, and the order of reactivity in ssDNA was Ade-N1>Gua-N7>Cyt-N3>Ade-N3. The most reactive site in dsDNA was Ade-N3. The Gua-N7 and Ade-N3 adducts were hydrolytically labile. Ade-N7 adduct could not be identified in the hydrolysates of ssDNA or dsDNA. The adduct Gua-N7,N7, which consists of two units of Gua bound together with a unit derived from chlorambucil, is a cross-linking adduct, and it was detected in the hydrolysates of ssDNA and dsDNA. Also several other adducts were detected which could be characterized by spiking with previously isolated authentic adducts or tentatively by MS. The role of chlorambucil-DNA adducts on the cytotoxicity and mutagenity of 1 is also discussed.

Characterization of adducts formed in reactions of acrolein with thymidine and calf thymus DNA.

Acrolein, an important industrial chemical and environmental contaminant, has been shown to interact with nucleic acids in vitro and in vivo. In this study, we examined the reactivity of acrolein towards thymidine and calf-thymus double- and single-stranded DNA in aqueous buffered solutions. LC-MS Analyses of the reaction mixture of acrolein with thymidine showed the formation of five structurally different adducts. The structures of the products were determined on the basis of mass spectrometry, UV absorbance, and (1)H- and (13)C-NMR spectroscopy. The adducts were identified as 3-(3-oxopropyl)thymidine (dT1), 3-[(tetrahydro-2,4-dihydroxypyran-3-yl)methyl]thymidine (dT2), 2-(hydroxymethyl)-5-(thymidin-3-yl)pent-2-enal (dT3), 3-hydroxy-2-methylidene-5-(thymidin-3-yl)pentanal (dT4), and 2-[(thymidin-3-yl)methyl]penta-2,4-dienal (dT5). The adducts dT2-dT5 were formed in reaction of dT1 with acrolein. In the reaction of acrolein with calf-thymus DNA, dT1 was the only adduct detected in the DNA hydrolysate.

Formation of adducts in the reaction of glyoxal with 2'-deoxyguanosine and with calf thymus DNA.

The reactions of glyoxal with 2'-deoxyguanosine and calf thymus single- and double-stranded DNA in aqueous buffered solutions at physiological conditions resulted in the formation of two previously undetected adducts in addition to the known reaction product 3-(2'-deoxy-beta-D-erythro-pentofuranosyl)-5,6,7-trihydro-6,7-dihydroxyimidazo[1,2-a]purine-9-one (Gx-dG). The adducts were isolated and purified by reversed-phase liquid chromatography and structurally characterised by UV absorbance, mass spectrometry, (1)H and (13)C NMR spectroscopy. The hitherto unknown adducts were identified as: 5-carboxymethyl-3-(2'-deoxy-beta-D-erythro-pentofuranosyl)-5,6,7-trihydro-6,7-dihydroxyimidazo[1,2-a]purine-9-one (Gx(2)-dG) and N(2)-(carboxymethyl)-9-(2'-deoxy-beta-D-erythro-pentofuranosyl)-purin-6(9H)-one (Gx(1)-dG). Both adducts were shown to arise from Gx-dG. Gx-dG and Gx(2)-dG were found to be unstable and partly transformed to Gx(1)-dG, which is a stable adduct and seems to be the end-product of the glyoxal reaction with 2'-deoxyguanosine. All adducts formed in the reaction of glyoxal with 2'-deoxyguanosine were observed in calf thymus DNA. Also in DNA, Gx(1)-dG was the only stable adduct. The transformation of Gx-dG to Gx(1)-dG seemed to take place in single-stranded DNA and therefore, Gx(1)-dG may be a potentially reliable biomarker for glyoxal exposure and may be involved in the genotoxic properties of the compound.

Reactions of malonaldehyde and acetaldehyde with calf thymus DNA: formation of conjugate adducts.

Our previous work has shown that treatment of nucleosides with malonaldehyde simultaneously with acetaldehyde affords stable conjugate adducts. In the present study we demonstrate that conjugate adducts are also formed in calf thymus DNA when incubated with the aldehydes. The adducts were identified in the DNA hydrolysates by their positive ion electrospray MS/MS spectra, by coelution with the 2'-deoxynucleoside standards, and, in the case of adducts exhibiting fluorescent properties, also by LC using a fluorescence detector. In the hydrolysates of double-stranded DNA (ds DNA), two deoxyguanosine and two deoxyadenosine conjugate adducts were detected and in single-stranded DNA (ss DNA) also, the deoxycytidine conjugate adduct was observed. The guanine base was the major target for the malonaldehyde-acetaldehyde conjugates and 2'-deoxyguanosine adducts were produced in ds DNA at levels of 100-500 adducts/10(5) nucleotides (0.7-3 nmol/mg DNA).

Formation of malonaldehyde-acetaldehyde conjugate adducts in calf thymus DNA.

It has previously been shown that malonaldehyde forms conjugates with acetaldehyde and that these conjugates react with nucleobases forming so-called conjugate adducts. In the current study, it is shown that conjugate adducts are also formed in calf thymus DNA when incubated simultaneously with malonaldehyde and acetaldehyde. The adducts were identified in the DNA hydrolysates by their positive ion electrospray MS/MS spectra and by coelution with the 2'-deoxynucleoside standards and, in the case of adducts exhibiting fluorescent properties, also by LC using a fluorescence detector. In the hydrolysates of double-stranded DNA (ds DNA), two deoxyguanosine and two deoxyadenosine conjugate adducts were detected, and in single-stranded DNA (ss DNA) also, the deoxycytidine conjugate adduct was observed. The guanine base was the major target for the malonaldehyde-acetaldehyde conjugates, and 2'-deoxyguanosine adducts were produced in ds DNA at levels of 100-500 adducts/10(5) nucleotides (0.7-3 nmol/mg DNA). The 2'-deoxyadenosine adducts and the 2'-deoxycytidine adduct were generated in higher amounts when the incubation was performed at pH 6.0 than at pH 7.4, while the opposite formation profile was noted for the 2'-deoxyguanosine adducts, especially in the ss DNA reaction. This observation is exactly in accordance with our previously reported pH-dependent reactivity of the individual nucleosides with malonaldehyde-acetaldehyde conjugates. The findings of this work show that the genotoxic effects observed for malonaldehyde and acetaldehyde could be in part due to the formation of the conjugate adducts.

Formation of acrolein adducts with 2'-deoxyadenosine in calf thymus DNA.

Acrolein is a ubiquitous environmental contaminant that has been found to be mutagenic in prokaryotic and eukaryotic cells. In the present study, we examined the reactions of acrolein with 2'-deoxyadenosine and calf thymus single- and double-stranded DNA in aqueous buffered solutions at physiological conditions. The deoxynucleoside adducts were isolated by reversed-phase liquid chromatography, and their structures were determined by their UV absorbance, mass spectrometry, and 1H and 13C NMR spectroscopy. The reaction of 2'-deoxyadenosine with acrolein resulted in the formation of four structurally different adducts (dAI, dAII, dAIII, dAIV). The structures of the novel acrolein adducts, dAIII and dAIV, were assigned as 3-[N(6)-(2'-deoxyadenosinyl)]propanal (dAIII) and 9-(2'-deoxyribosyl-6-(3-formyl-1,2,5,6-tetrahydropyridyl)purine (dAIV), respectively. The adduct dAIII was found to arise via a Dimroth rearrangement of adduct dAI, while the adduct dAIV was shown to be formed upon further reaction of acrolein with dAIII. In the reaction of acrolein with calf thymus DNA, all studied 2'-deoxyadenosine-acrolein adducts were observed. For the first time, it is shown that the N(6)-adduct and the adducts which are derived from two acrolein units are formed in calf thymus DNA.

Reaction of acrolein with 2'-deoxyadenosine and 9-ethyladenine--formation of cyclic adducts.

Treatment of 2'-deoxyadenosine with acrolein at pH 4.6 in 37 degrees C affords unstable adducts containing either one or two fused ring systems where the hydroxypropano units are derived from acrolein. Since the use of 2'-deoxyadenosine resulted in the creation of at least four diastereoisomers for the adduct made up of two fused rings, therefore, for identification and assignment of the products, 9-ethyladenine was used instead as the starting material in the reaction. The products, 3E and 4E, were structurally characterised by UV, mass spectrometry and NMR spectroscopy.

Evaluation of millers' dental health. Part II. State of the parodontium.

A study of the oral health of workers in flour mills was carried out. The examined group consisted of 40 males and 8 females currently employed at flour mills. As much as 97.92 % of the workers showed evidence of parodontopathy. The results of the researches indicate the necessity of professional periodontal prophylaxis among mill workers.