Nos3 Gene Rs1799983 and Rs2070744 Polymorphisms in Patients with Periodontal Disease.

Periodontal disease is a common oral disease. Inflammatory and immune responses to oral microorganisms initiate the development of periodontitis. Cigarette smoking is an important environmental risk factor for periodontitis. Another important inflammatory mediator is nitric oxide (NO). NO modulates vascular tone, microvascular permeability, leukocyte migration and oxidative activity, contributing to the direct killing of microorganisms. Several polymorphisms of the NOS3 gene have been detected, which may alter gene expression and NO synthesis. The aim of this study was to examine the association between the NOS3 rs1799983 and rs2070744 polymorphisms and periodontal disease. This study enrolled 200 patients with periodontal diseases (130 were non-smokers and 70 were smokers) and 160 control subjects (126 were non-smokers and 34 were smokers). Among the patients with periodontal disease, we observed a statistically increased frequency of patients with the CT genotype (TC vs. TT; 95%CI 1.83, OR 1.16-2.88, P = 0.011). There was a statistically significant increased frequency of CT genotype carriers among non-smoking patients with periodontal disease as compared with non-smoking controls, whereas there were no statistically significant differences between smoking patients with periodontal disease and smoking control subjects. The results of our study suggest an association between the NOS3 rs2070744 polymorphism and periodontal disease.

Association of the 174 G/C polymorphism of the IL6 gene in Polish power-orientated athletes.

AIM: Interleukin-6 (IL6), has been called by some authors "an exercise factor" due to its pleiotropic effects during physical training. Several studies indicated that change in the guanine bases to cytosine at position -174 affects the transcription of the IL6 gene, and finally IL6 production level. The aim of this study was to perform confirmation studies to analyze the possible importance of the IL6 -174 G/C (rs1800795) polymorphism gene in Polish power-orientated athletes. METHODS: The study was carried out on two groups of men of the same Caucasian descent: 158 power-orientated athletes and 254 volunteers not involved in competitive sport. DNA was extracted from buccal cells donated by the subjects. Genotyping was carried out by polymerase chain reaction (PCR). Significance was assessed by Chi square (χ2) analysis. RESULTS: The results revealed that the frequency of the IL6 -174 GG genotype (53.16% vs. 35.82%; P=0.002) and G allele (68.67% vs. 57.87%; P=0.03) were significantly higher in the Polish power-orientated athletes compared to controls. CONCLUSION: These data suggest that the G allele could be one of the factors influencing the power-orientated sport performance. However, these conclusions should be supported with more experimental studies on other IL6 polymorphisms and other genes.

Recovery of DNA of Giardia intestinalis cysts from surface water concentrates measured with PCR and real time PCR.

The most important restriction for the detection in water samples is the low concentration of Giardia intestinalis cysts, additional difficulty is the presence of PCR inhibitors. We have carried out trials in order to assess the sensitivity of semi-nested PCR and TaqMan real time PCR on the basis of DNA extracted from G. intestinalis cysts coming from spiked environmental and distilled water samples, filtrated with the use of Filta-Max® equipment (1623 Method). Removal of inhibitors was carried out with addition of BSA in different concentrations. During the filtration and concentration of water samples, losses of cysts have been recorded. Moreover, addition of BSA to the PCR and real time PCR mix increases the sensitivity of reaction. The optimal concentration of BSA for semi­nested PCR was 15 and 20 ng/µl, whereas for real time PCR 5 ng/µl.

Comparison of efficiency of various DNA extraction methods from cysts of Giardia intestinalis measured by PCR and TaqMan real time PCR.

The aim of the presented study was to work out an effective method of extraction of DNA from Giardia intestinalis cysts as well as a sensitive and specific method for detection of DNA of this protozoan using a polymerase chain reaction (PCR). Twelve protocols for DNA extraction have been compared. Purification and extraction of DNA were preceded by additional actions in order to destroy the cysts' wall. The highest effectiveness of DNA extraction was obtained in case of alternating application of freezing the samples in liquid nitrogen and their incubation in water bath in the temperature of 100 degrees C, and then the extraction with the QIAamp DNA Tissue Mini Kit (QIAGEN)--T kit--with an all night long incubation with proteinase K in 56 degrees C. Effectiveness of DNA extraction with the use of each kit after extraction with each treatment was measured by nested PCR product of beta-giardin gene fragment and C(T) values of real time PCR of the SSU rRNA gene of G. intestinalis. The detection limit, defined as the lowest number detected in 100% cases, was 100 cysts per 200 microl when effectiveness was evaluated with nested PCR and 50 oocysts with real time PCR after extraction DNA with T kit. Results of our comparative studies have shown that all stages preceding the molecular detection of G. intestinalis DNA are equally important, and materially influence on the final effect and this version of method seems to be very useful for the sensitive detection of DNA of G. intestinalis.

The angiotensin converting enzyme gene I/D polymorphism in Polish rowers.

Angiotensin converting enzyme gene (ACE) is the most frequent investigated gene in the context of genetic conditioning of sports-predispositions. Product of this gene is a key-element in the renin-angiotensin system responsible for the regulation of blood pressure. In this study DNA polymorphism in the ACE gene was studied in Polish rowers in order to examine the hypothesis that ACE genotype is associated with athletes performance. Fifty-five male Polish rowers including Olympic and World champions were recruited for this study. Control samples were prepared from 115 unrelated volunteers. PCR amplification of the insertion (I) or deletion (D) fragment of ACE gene was performed. Genotype distribution and allele frequencies were determined by genotype and gene counting. Significance was assessed by chi2 analysis. ACE genotype distributions amongst subjects and controls were in Hardy-Weinberg equilibrium. Compared with controls, the frequency of I allele differ significantly from that found in rower's group: 56.3% vs. 44.3%, (P=0.038) and ACE genotype frequency amongst the whole athletes group (30.9% II, 50.9% ID, 18.2% DD) was also different from expected values (control group 19.1% II, 50.4% ID, 30.4% DD; P=0.039). This data confirm a positive association of the I allele of ACE gene with endurance performance.

Galanin influences vasopressin and oxytocin release from the hypothalamo-neurohypophysial system of salt loaded rats.

Galanin is a peptide present in the nervous system and peripheral tissues which exerts a broad range of physiological functions. The influence of centrally administered galanin (Gal; 100 pM i.c.v.) on arginine vasopressin (AVP) and oxytocin (OT) content in the hypothalamus and neurohypophysis as well as on their blood plasma concentration was estimated in male Wistar rats drinking ad libitum 2% solution of natrium chloride per 48 hours. In euhydrated rats and subsequently applied i.c.v. with Gal a significant fall in the hypothalamic and neurohypophysial content of OT but not AVP was observed, however, without simultaneous changes in these neurohormones blood plasma concentration. On the contrary, i.c.v. injection of Gal to salt-loaded rats caused a marked raise in AVP and OT level in the hypothalamus and neurohypophysis with subsequent diminution of both neurohormones concentration in blood plasma. These results suggest that in euhydrated rats Gal has an inhibitory influence on the biosynthesis as well as axonal transport of OT, but not AVP. On the contrary, in salt-loaded rats galanin restricts secretion of both neurohormones into the systemic circulation.

[Population genetics of 9 Y-chromosome STR loci w Northern Poland]. / Genetyka populacyjna 9 loci typu Y-STR w populacji Polski Pólnocnej.

A total of 508 unrelated males from the North Poland population were analyzed for 9 Y-chromosome STRs (DYS 19, DYS 390, DYS 393, DYS 392, DYS 391, DYS 389I, DYS 389II and DYS 385I/II) using two multiplex reactions and detection of PCR products using capillary electrophoresis. In the analyzed sample 328 different haplotypes were identified, among which 264 were unique. It was found that the model for a Polish population haplotype (DYS 19*17, DYS 390*25, DYS 393*13, DYS 392*11, DYS 391*10, DYS 389I*13, DYS 389II*30, DYS 385I/II*10,14) is almost 15 times more frequent in our population than in a cumulative European one. The haplotype diversity/discrimination index calculated for 9 loci is 0.9943. In the analysed population sample three mutations were detected in the DYS19 (duplication) and DYS385I/II loci (triplications).

[Application of pcr in paternity determination in pregnancy preceded by rape]. / Zastosowanie reakcji lancuchowej polimerazy (PCR) we wczesnej ciazy w badaniu spornego ojcostwa zwiazanego z gwaltem.

Authors present a case of PCR (Polymerase Chain Reaction) technique application for the paternity determination in pregnancy preceded by the rape. Diagnostic material was obtained in the 10th week of pregnancy by the use of transabdominal Chorionic Villous Sampling under the echo-guidance and its DNA feature was compared against the DNA material obtained from the vagina after the rape and the material from mother and her husband. The PCR technique revealed in chorionic villi the presence of allele that were present in material deriving from mother and her husband as well as no even single allele from the material of violator. Exclusion of violator as a father of the foetus effected in decision of continuing the pregnancy.

[Free crystalline silica: possibilities and limitations of its detection and determination in dust using infrared spectrometry]. / Wolna krystaliczna krzemionka--mozliwosci i ograniczenia wykrywania oraz oznaczania w pylach metoda fourierowskiej spektrometrii w podczerwieni (FT-IR).

The effect of the commonly occurring industrial dust components on the detection of crystalline silica and crystobalite by means of infrared spectrometry was investigated. The influence of minerals interfering in infrared with quartz and crystobalite: kaolinite, microcline, albite, mullite, and amorphous silica, as well as of the mixture of SiO2 crystalline forms was analyzed. The study was based on the computer-aided summing up of crystalline silica spectra with spectra of interfering components and spectra resulting from further transformation: deconvolved spectra, the second and fourth orders derivative spectra. It was found that the smallest amount of silica, possible to be detected in mixtures with interfering components amounted to 1-5% and crystobalite to 2-10%. The limit of detection, using the pellet technique in the adopted conditions of spectra registration, accounted for 0.5 microgram for silica and 1 microgram for crystobalite, whereas using PVC filters, 1 microgram of silica and 4 micrograms of crystobalite could be detected.

Solid dispersion of ketoprofen in pellets.

The formulation of ternary solid dispersions of ketoprofen with Macrogol and kollagen hydrolizate derivative as carriers was elaborated on the basis of the results of the experiments in which different methods of solid dispersion preparation (melting, solvent method, different cooling), different concentrations of drug/carriers and molecular weight of Macrogol were tested. The best solid dispersion consisted of: ketoprofen-Macrogol 6000-KLH(T) (1+8. 9+0.1) was chosen to formulate the pellets on the basis of the pharmaceutical availability of ketoprofen from solid dispersion and the physical chemical studies: thermomicroscopic, DSC and X-ray diffraction. The pellets were prepared by the extrusion and spheronization method. The mechanical properties of the pellets as well as ketoprofen released from pellets containing solid dispersion, in comparison with physical mixtures and the drug alone, were evaluated. The increase in the amount of released ketoprofen from solid dispersion pellets was 3.8-times greater than from the pellets containing the drug alone. The stability of solid dispersion pellets was satisfactory.

Forensic validation of a multiplex containing nine STRs--population genetics in northern Poland.

This paper presents the allele frequency distributions for the nine loci (D3S1358, VWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820) present in the commercially available Profiler Plus kit. DNA samples of 202 individuals from Northern Poland were amplified in a multiplex reaction with subsequent automatic detection using capillary electrophoresis. All loci met the Hardy-Weinberg equilibrium conditions. The calculated probability of identity was 2.26 x 10(-11) giving an average probability of identity of 1 in 44 billion. Considerable improvement of analysis precision was observed after substituting the GeneScan 500 for the fluorescent ladder 60-400 bp. The influence of DNA concentration on stutter and artefact formation and the ratio of heterozygote alleles was analysed.

Template-directed base pairing of 2-chloro-2'-deoxyadenosine catalyzed by AMV reverse transcriptase.

2-Chloro-2'-deoxyadenine (2CldA) is used for treatment of several lymphoid malignancies. Since this drug is incorporated into DNA, we have undertaken studies on base pairing of 2-chloroadenine (2ClA). 2CldA phosphoramidite was synthesized and used for preparation of 25-mer templates with 2ClA located at site 21 from the 3'-end. Kinetic parameters (Km and Vmax) for the incorporation of deoxynucleoside-5'-triphosphates by AMV reverse transcriptase opposite the 2ClA template, as well as for the extension of 2ClA.T pair, were determined. The efficiency (Vmax/Km) of incorporation of dGTP, dCTP, and dATP opposite 2ClA is at least one order of magnitude lower than opposite unmodified A. The efficiency of incorporation of dTTP opposite 2ClA is about 30-fold lower than opposite A and extension of 2ClA.T pair is 3-fold lower than of A.T pair. From the analysis of the parameters of dTTP incorporation we conclude that formation of 2ClA.T pair is thermodynamically, but not kinetically controlled. The difference in binding energy (deltadeltaG) between 2ClA.T and A.T pairs in the environment of the polymerase active site is 2 kcal/mol. Our results indicate that the presence of 2ClA in DNA slows down replication, but does not lead to base-substitution mutations.

[Exposure to ceramic fibers in the work environment. III. occupational exposure to ceramic fibers in plants which produce and apply insulation materials made of ceramic fibers]. / Narazenie na wlókna ceramiczne w srodowisku pracy. III. Zawodowa ekspozycja na wlókna ceramiczne w zakladach produkujacych i stosujacych materialy izolacyjne z wlókien ceramicznych.

The study was aimed at assessing the exposure to dust in the work environment of plants which produced and apply packing and insulation materials made of ceramic fibres. The study revealed that workers were exposed to dust containing respirable ceramic fibres and in some cases (production of packing cord, insulating tape and paperboard) respirable asbestos fibres. The mean concentration of respirable fibres ranged from 0.05 to 0.62 f/l cm3, and concentrations of total dust fell between 0.6 and 23.2 mg/m3. The mean concentrations of respirable fibres were usually below (0.5 f/l cm3 for respirable ceramic fibres with asbestos mixture; 1 f/cm3 for respirable ceramic fibres), and of total dust much higher (1 mg/m3 and 2 mg/m3, respectively) than MAC values proposed. The initial dermatological examinations (25 workers) allow the conclusion that contact with ceramic fibres induces in some workers acute dermatitis and dermal papilla.

Frequencies for five short tandem repeat (STR) systems in a population from north Poland.

A population study of unrelated individuals from North Poland (Gdansk area) was carried out to investigate the allele distributions of the five STR systems HUMCD4, HUMFES/FPS, HUMVWA31, HUMTH01 and ACTBP2. PCR products were separated on horizontal non-denaturing polyacrylamide gels followed by silver staining. For all STR systems analysed the distribution of observed phenotypes did not deviate from Hardy-Weinberg equilibrium. A comparison of allele distributions between Polish and other European Caucasian population samples is presented.

[Biological effect of wood dust]. / Biologiczne dzialanie pylu drzewnego.

The biological effect of exposure to wood dust depends on its composition and the content of microorganisms which are an inherent element of the dust. The irritant and allergic effects of wood dust have been recognised for a long time. The allergic effect is caused by the wood dust of subtropical trees, e.g. western red cedar (Thuja plicata), redwood (Sequoia sempervirens), obeche (Triplochiton scleroxylon), cocabolla (Dalbergia retusa) and others. Trees growing in the European climate such as: larch (Larix), walnut (Juglans regia), oak (Quercus), beech (Fagus), pine (Pinus) cause a little less pronounced allergic effect. Occupational exposure to irritative or allergic wood dust may lead to bronchial asthma, rhinitis, alveolitis allergica, DDTS (Organic dust toxic syndrome), bronchitis, allergic dermatitis, conjunctivitis. An increased risk of adenocarcinoma of the sinonasal cavity is an important and serious problem associated with occupational exposure to wood dust. Adenocarcinoma constitutes about half of the total number of cancers induced by wood dust. An increased incidence of the squamous cell cancers can also be observed. The highest risk of cancer applies to workers of the furniture industry, particularly those dealing with machine wood processing, cabinet making and carpentry. The cancer of the upper respiratory tract develops after exposure to many kinds of wood dust. However, the wood dust of oak and beech seems to be most carcinogenic. It is assumed that exposure to wood dust can cause an increased incidence of other cancers, especially lung cancer and Hodgkin's disease. The adverse effects of microorganisms, mainly mould fungi and their metabolic products are manifested by alveolitis allergica and ODTS. These microorganisms can induce aspergillomycosis, bronchial asthma, rhinitis and allergic dermatitis.

[Evaluation of occupational exposure of workers engaged in mining of rock raw materials in the Sleza massif]. / Ocena narazenia zawodowego pracowników zatrudnionych przy wydobyciu surowców skalnych na obszarze masywu Slezy.

In two serpentinite mines and in a factory producing asphalt pavements using serpentinite, exposure to dusts and polycyclic aromatic hydrocarbons (WWA-fraction soluble in benzene) was assessed. In rock samples taken in the serpentinite mine and in dust samples from work posts crystalline phases were determined (using X-ray radiography and spectrometry in infrared). Also, morphology of dust particles was performed using electron microscopy. Both in the rock samples and in dust fibrous antigorite, a mineral of strong cancerogenic properties, was found. Total dust concentrations varied between 1.05 and 280 mg/m3, concentration of respirable mineral fibres from 0.01 to 3.56 fibre/cm3, and concentration of aromatic hydrocarbons varied between 266.7 and 850 micrograms/m3. The study will be continued concerning evaluation of biological effect of serpentinite in animal experiments and assessment of extraoccupational exposure of people living in the neighbourhood of serpentinite mines and plants processing this mineral.

[Biological effect of cement dust]. / Biologiczne dzialanie pylu cementowego.

The harmful effect of cement dust upon living organisms consists in irritating, sensitizing and pneumoconiotic properties of its components. In animal studies it has been observed that cement dust induces atrophic and hypertrophic changes in nasal and pharyngeal mucosa and chronic exfoliative bronchitis. In the lungs of experimental animals slight tissue fibrosis and some emphysema foci were found. The examination of workers exposed to cement dust has shown that disorders of the upper respiratory airways they suffer from include most often chronic rhinitis, laryngitis and pharynx catarrh. Also, it has been noticed that chronic bronchitis in the exposed workers was 1.7 times more frequent compared to those non-exposed, and that asthma was diagnosed in some of the exposed workers. Chronic bronchitis was usually characterized by the symptoms of impaired, obstructive lung ventilation. Defects in lung ventilation were strictly related to the duration of mild cases of cement pneumoconiosis diagnosed in a small percentage of workers who were exposed for at least 10 years to high dust concentrations. Long-term contact of skin with cement results in inflammatory changes or, in some cases, in chemical burns. Etiological factors of inflammatory skin changes are allergenic elements (Cr, Nr, Co) and irritating agents found in cement.

[Experimental silicosis. Analysis of collagen levels in the heart, spleen and liver of rats in experimental silicosis]. / Doswiadczalna pylica krzemowa. Analiza poziomu kolagenu w sercu, sledzionie i watrobie szczurów z krzemica doswiadczalna.

Possibilities of increasing the content of collagen in heart, spleen and liver have been tested in rats affected by silicosis induced by intratracheal administration of 50 mg of siliceous earth dust suspension. An increased level of collagen has been found in rats' hearts after 9 months, whereas in spleens after 6 and 9 months of experiment. In the liver of rats suffering from experimental silicosis, no changes which would clearly indicate the fibrosis of this organ were found. In addition, the animals administered with dust exhibited inhibited physiological growth of the body weight and liver weight.

[Incidence of pneumoconiosis and the fibrogenic properties of dust occurring during the production of thermallite bricks]. / Zachorowalnosc na pylice pluc i wlasciwosci zwlókniajace pylów wystepujacych podczas produkcji cegly termalitowej.

Notifications of occupational diseases among building ceramics industry workers were analysed. During 1976-1980 six cases of pneumoconiosis among those workers were diagnosed. All those cases referred to workers of two divisions dealing with transport and preparation of raw materials for the production of thermallite brick of a mixture of clay from Jaroslaw, siliceous earth from Piotrowice and sawdust. Experiments on animals confirmed a very high biological aggressiveness of unheated siliceous earth and weak fibrogenic activity of clay dusts and dusts emitting from the burned thermallite brick. Diffractometric tests of the dusts indicated that the high biological aggressiveness of siliceous earth was due to alpha-crystoballite of low crystallinity. Under effect of high temperature (during brick burning) crystoballite of siliceous earth is subject to great changes (the degree of crystallinity gets increased), which results in its decreased fibrogenic properties. The experiments on animals confirmed a weak biological aggressiveness of the dust from burnt thermallite brick and considerably decreased aggressiveness of siliceous earth heated at 1200 degrees C for 4 hours, as compared to unheated siliceous earth. The weak fibrogenic activity of the dust of clay containing approx. 15% of alpha-quartz may be due to kaolinite and illite in this dust.