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The implementation of new methods of osteoporotic therapy requires tests on animal model. The use of sheep as model has numerous advantages over other animals. The aim of this study was to describe the change in parameters in sheep with osteoporosis induced using steroids and ovariorectomy methods as opposed to the parameters in healthy sheep. The study was performed on female "merinos" breed sheep divided into the three groups: negative control (NC)--healthy animals, positive control (PC)--ovariorectomized animals and steroid control group (SC)--in which methylprednisolone was administered. This paper presents histological and ultrastructural examination with mechanical comparative tests for force/strength values as well as indentation tests of joint cartilage. The obtained results confirm the loss of bone mass associated with mineral composition content in bones, which has an influence on bone strength.

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The implementation of new methods of oseoporotic therapy requires tests on an animal model. One of the best is the sheep, whose numerous advantages over other models are described in the literature. The aim of this study was induction of osteoporosis using steroids and ovariorectomy methods in sheep and description of the change in parameters with regard to healthy sheep. The study was performed on female "merino" breed sheep divided into three groups: Negative control (NC) healthy animals, positive control (PC) ovariorectomised animals and steroid group (SC) where methylprednisolone was implemented. Blood tests, diagnostic arthroscopy, quantitative computed tomography and X-Ray micro-tomography of bone were carried out. Blood tests revealed a decreased level of estrogens, progesterone and increased parathormone and cortisol levels in the SC group. A decrease in bone turnover markers and an increase in bone resorption markers in all groups were also noted. Diagnostic arthroscopy revealed osteoarthrosis in PC and SC groups. Radiological density tests showed a slight decrease in PC and NC groups whereas there was more than a triple decrease in SC. Results obtained from microCT showed quickly developing osteoporosis in the SC group, which is reflected in numerous parameters analysed in this study. The best effects for osteoporosis induction were obtained using ovariorectomised sheep with methylprednisolone injections.

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Osteochondrosis is a common and clinically important joint disorder that occurs in human and many species of animals such as pigs, horses and dogs. The aim of this article is to present the results of conservative and surgical treatment of Osteochondrosis dissecans (OCD) and to compare the recovery time in relation to race, age and extent of cartilage damage. The study was performed on 36 dogs of both sexes, aged from 4 to 24 months, weighing 12-70 kg, of different breeds (predisposed and non-specific for this disease). All patients underwent clinical, orthopedic and radiological examination through which the location of OCD changes in the shoulder joint and the degree of this problem were established. Based on the examinations the patients were qualified for particular treatment procedures. Control tests were performed after 14, 30 and 60 days, which allowed for a precise evaluation of the progress of therapy. Achieved results indicate that dogs age and the lesions extent in the shoulder joint have an impact on the outcome of treatment. Studies showed that the best therapeutic effects were obtained in dogs aged 6-10 months, which underwent surgery. The study proved that the early diagnosis of OCD strongly influence the recovery rate and may inhibit the development of degenerative changes in the joint (DJD).

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The nanosystem composed of only as few as seven fullerenols C(60)(OH)(24) molecules was studied using the molecular dynamics (MD) technique. The interaction was taken to be the full site-site pairwise additive Lennard-Jones (LJ) potential, which generates both translational and anisotropic rotational motions of each fullerenol. The radial distribution function, the mean square displacement, the Lindemann index, the translational and angular velocity correlation functions of fullerenol have been calculated for several energies of the nanosystem. We provide also the in-depth evidence of the solid-liquid phase transition in the investigated cluster. The comparison with the properties of pure fullerene cluster (C(60))(7) is provided. The study are motivated by the expected diverse biological applications of water-soluble fullerenols.

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OBJECTIVE: To determine which factors were associated with hospitalization for tuberculosis (TB) in New York City (NYC), United States, and to estimate the proportion of potentially avoidable admissions. DESIGN: Patients diagnosed with TB from April to June 2003 were included. Records of hospitalized patients were reviewed to determine whether hospitalization was appropriate. Hospitalization was considered appropriate if patients met ≥1 of the NYC health department hospitalization criteria and/or needed hospitalization per study physicians' judgment. The association of patient characteristics with hospitalization and with having an inappropriate hospitalization was evaluated using multivariate analyses. TB cases from 2008 were also evaluated to determine whether more recent cases had similar associations with hospitalization. RESULTS: Of 315 patients diagnosed with TB during the study, 226 (72%) were hospitalized. Hospitalized patients were more likely to have a cavitary chest radiograph (adjusted odds ratio [aOR] 8.11, 95%CI 1.82-36.20), abuse alcohol/drugs (aOR 6.53, 95%CI 2.06-20.67), be Black non-Hispanic (aOR 3.05, 95%CI 1.00-9.38), have unknown human immunodeficiency virus status (aOR 2.67, 95%CI 1.24-5.76), and to have been first evaluated by a private medical provider (aOR 2.37, 95%CI 1.11-5.08). Eighty-seven (38%) of the hospitalizations may have been inappropriate; foreign-born (aOR 3.16, 95%CI 1.39-7.14) and acid-fast bacilli sputum smear-positive (aOR 2.49, 95%CI 1.18-5.23) patients were more likely to be hospitalized inappropriately. CONCLUSION: Many TB hospitalizations in NYC may be avoidable. Existing guidelines for diagnosing and managing cases as out-patients need to be put into practice.

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OBJECTIVE: Onconeuronal antibodies are important diagnostic tool in patients with suspicion of paraneoplastic neurological syndromes (PNS). However, their role in PNS pathophysiology and specificity for particular neurological manifestation remains unclear. The aim of this study was to evaluate onconeuronal and antineuronal antibodies in patients with pulmonary pathologies and suspected for PNS. MATERIAL AND METHODS: Twenty one patients with pulmonary pathologies were selected from the database of 525 consecutive patients with suspicion of PNS. Patients' sera were screened for the presence of onconeuronal and antineuronal antibodies by means of indirect immunofluorescence; the presence was confirmed by Western blotting. Clinical data were obtained from medical records, hospital data base, and questionnaire-based direct telephone contact with patients. RESULTS: Among 21 patients, aged 54 +/-11, with pulmonary pathologies, the most frequent neurological manifestations were neuropathies. Typical PNS included paraneoplastic cerebellar degeneration (PCD) and limbic encephalitis (LE). We found cases with multiple onconeuronal antibodies (anti-Ri and anti-Yo) and coexisting PNS (PCD/LE). Well-defined onconeuronal antibodies were identified in 23.8% of patients. Among antineuronal antibodies, the most frequent were anti-MAG (23.8%). ROC curves analysis revealed high sensitivity of onconeuronal and antineuronal antibodies for typical PNS and lower for pulmonary malignancies. CONCLUSIONS: Tests for antibodies are highly sensitive for the diagnosis of typical paraneoplastic neurological syndromes. Anti-myelin and anti-MAG antibodies are associated with non-neoplastic pulmonary diseases. Patients with well-defined onconeuronal antibodies require careful screening and follow-up, because the PNS diagnosis indicates a high probability of an underlying malignancy.

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We have simulated (by the molecular dymanics (MD) method) the dynamics of fullerenes (C(60)) in an extremely small cluster composed of only as many as seven C(60) molecules. The interaction is taken to be the full 60-site pairwise additive Lennard-Jones (LJ) potential which generates both translational and anisotropic rotational motions of each molecule. Our atomically detailed MD simulations discover the plastic phase (no translations but active reorientations of fullerenes) at low energies (temperatures) of the (C(60))(7) cluster. We provide the in-depth evidence of the dynamical solid-liquid bistability region in the investigated cluster. Moreover, we confirm the existence of the liquid phase in (C(60))(7), the finding of Gallego et al (1999 Phys. Rev. Lett. 83 5258) obtained earlier on the basis of Girifalco's model, which assumes single-site only and spherically symmetrical interaction between C(60) molecules. We have calculated the translational and angular velocity autocorrelation functions and estimated the diffusion coefficient of fullerene in the liquid phase.

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Past genotypic studies of Mycobacterium tuberculosis may have incorrectly estimated the importance of specific drug resistance mutations due to a number of sampling biases including an overrepresentation of multidrug-resistant (MDR) isolates. An accurate assessment of resistance mutations is crucial for understanding basic resistance mechanisms and designing genotypic drug resistance assays. We developed a rapid closed-tube PCR assay using fluorogenic reporter molecules called molecular beacons to detect reportedly common M. tuberculosis mutations associated with resistance to isoniazid and rifampin. The assay was used in a comparative genotypic investigation of two different study populations to determine whether these known mutations account for most cases of clinical drug resistance. We analyzed samples from a reference laboratory in Madrid, Spain, which receives an overrepresentation of MDR isolates similar to prior studies and from a community medical center in New York where almost all of the resistant isolates and an equal number of susceptible controls were available. The ability of the molecular beacon assay to predict resistance to isoniazid and rifampin was also assessed. The overall sensitivity and specificity of the assay for isoniazid resistance were 85 and 100%, respectively, and those for rifampin resistance were 98 and 100%, respectively. Rifampin resistance mutations were detected equally well in isolates from both study populations; however, isoniazid resistance mutations were detected in 94% of the isolates from Madrid but in only 76% of the isolates from New York (P = 0.02). In New York, isoniazid resistance mutations were significantly more common in the MDR isolates (94%) than in single-drug-resistant isolates (44%; P < 0.001). No association between previously described mutations in the kasA gene and isoniazid resistance was found. The first mutations that cause isoniazid resistance may often occur in sequences that have not been commonly associated with isoniazid resistance, possibly in other as yet uncharacterized genes. The molecular beacon assay was simple, rapid, and highly sensitive for the detection of rifampin-resistant M. tuberculosis isolates and for the detection of isoniazid resistance in MDR isolates.

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Discovery of genotypic markers associated with increased transmissibility in Mycobacterium tuberculosis would represent an important step in advancing mycobacterial virulence studies. M. tuberculosis strains may be classified into one of three genotypes on the basis of the presence of specific nucleotide substitutions in codon 463 of the katG gene (katG-463) and codon 95 of the gyrA gene (gyrA-95). It has previously been reported that two of these three genotypes are associated with increased IS6110-based clustering, a potential proxy of virulence. We designed a case-control analysis of U.S.-born patients with tuberculosis in San Francisco, Calif., between 1991 and 1997 to investigate associations between katG-463 and gyrA-95 genotypes and epidemiologically determined measures of strain-specific infectivity and pathogenicity and IS6110-based clustering status. We used a new class of molecular probes called molecular beacons to genotype the isolates rapidly. Infectivity was defined as the propensity of isolates to cause tuberculin skin test conversions among named contacts, and pathogenicity was defined as their propensity to cause active disease among named contacts. The molecular beacon assay was a simple and reproducible method for the detection of known single nucleotide polymorphisms in large numbers of clinical M. tuberculosis isolates. The results showed that no genotype of the katG-463- and gyrA-95-based classification system was associated with increased infectivity and pathogenicity or with increased IS6110-based clustering in San Francisco during the study period. We speculate that molecular epidemiologic studies investigating clinically relevant outcomes may contribute to the knowledge of the significance of laboratory-derived virulence factors in the propagation of tuberculosis in human communities.

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We developed a new approach to DNA sequence analysis that uses fluorogenic reporter molecules--molecular beacons--and demonstrated their ability to discriminate alleles in real-time PCR assays of genomic DNA. A set of overlapping molecular beacons was used to analyze an 81-bp region of the Mycobacterium tuberculosis rpoB gene for mutations that confer resistance to the antibiotic rifampin. In a blinded study of 52 rifampin-resistant and 23 rifampin-susceptible clinical isolates, this method correctly detected mutations in all of the resistant strains and in none of the susceptible strains. The assay was carried out entirely in sealed PCR tubes and was simple to perform and interpret. This approach can be used to analyze any DNA sequence of moderate length with single base pair accuracy.

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Within the group of 260 members of medical staff, the presence of the HBV markers was found in 87 persons (33.5%). Vaccination, which was of a voluntary character, was initiated in the group of 75 persons according to the pattern of 0, 1, 6 months with the use of vaccine GenH-B-Vax (Merck Sharp Dohme-Chibret A. G.) was completed in 62 persons. Humoral response was investigated with quantitative measurement of anti-HBs 30 days after each doses of the vaccine. After three doses of vaccine a protective level of anti-HBs (greater than or equal to 10 IU/l) have been obtained in 90.3% of vaccinated. Humoral response was higher in the group of younger (below 35 years of life) employees. Investigated vaccine was highly immunogenic and was well tolerated.

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