Comparison study for genotyping of a single-nucleotide polymorphism in the tumor necrosis factor promoter gene.

A variety of methods that address the detection of single-nucleotide polymorphism (SNP) have been used in molecular diagnostics. The allele-specific polymerase chain reaction (ASPCR) has been one of the most extensively studied, including its application in the tumor necrosis factor (TNF)(-308) genotyping. Many studies have demonstrated that the ASPCR sensitivity and specificity depends on various PCR parameters, with mismatches occurring to a degree of 4%. The purpose of our study was to evaluate a comparison of genotyping of the TNF(-308) using an ASPCR and automated sequencing (ASEQ). In a total of 204 DNA samples, their duplicate examination by the ASPCR and ASEQ revealed concordant results in 96.5% and mismatches in 3.5% genotypes. Depending on the target TNF(-308G/G), TNF(-308G/A) , TNF(-308A/A) sequences, this translated into decreased ASPCR sensitivity to a degree of 98.6%, 94.2%, 60.0%, specificity 94.7%, 97.4%, 100.0%, positive predictive values 97.9%, 92.5%, 100.0%, and negative predictive values 96.4%, 98.0%, 99.0%, respectively. Based on these results, we found ASEQ to be more accurate than ASPCR for the TNF(-308) genotyping. By eliminating the need of empirical determination of appropriate PCR conditions for each studied sequence, ASEQ provides a sensitive and reproducible quality-control benchmark for other SNP assays.     

Initiation of protein synthesis and heat-shock protein-72 expression in the rat brain following severe insulin-induced hypoglycemia

Summary Following stress such as heat shock or transient cerebral ischemia, global brain protein synthesis initiation is depressed through modulation of eucaryotic initiation factor (eIF) activities, and modification of ribosomal subunits. Concomitantly, expression of a certain class of mRNA, heat-shock protein (HSP) mRNA, is induced. Here we report that the activity of eucaryotic initiation factor-2 (eIF-2), a protein that participates in the regulation of a rate-limiting initiation step of protein synthesis, transiently decreases following insulin-induced severe hypoglycemia in the rat brain neocortex. Expression of HSP 72, a 72-kDa HSP, in surviving neurons was seen at 1–7 days of recovery following 30 min of hypoglycemic coma, but not at 1 h and 6 h of recovery. In the neocortex, HSP 72 was first seen in layer IV, and later also in surviving neurons in layer II. In the CA1 region and in the crest of dentate gyrus, HSP 72 expression was evident in cells adjacent to irreversibly damaged neurons. In the CA3 region and the hilus of dentate gyrus, HSP 72 was expressed in a few scattered neurons. In septal nucleus, HSP 72 was expressed in a lateral to medial fashion over a period of 1–3 days of recovery. We conclude that severe insulin-induced hypoglycemia induces a stress response in neurons in the recovery phase, including inhibition of protein synthesis initiation, depression of eIF-2 activity, and a delayed and prolonged expression of HSP 72 in surviving neurons. The HSP 72 expression may be a protective response to injurious stress.     

Dislokation und ausscheidung des quecksilbers bei den ratten nach applikation von unitiol

Zusammenfassung In Laborverfahren wurde die Wirkung von Unitiol (2,3-Dimerkaptosulfonat-Natrium) auf die Quecksilberausscheidung aus dem Organismus beobachtet.Bei 16 weißen Ratten wurde Quecksilber in Einzeldosen von 0,2 mg/kg einmalig intravenös eingefüihrt. Dann verabreichte man den Ratten Unitiol in Einzeldosen von 5 mg/kg. Innerhalb von 6 Tagen bekam jedes Tier 11 mg Unitiol.Es wurde eine sichtbare Steigerung der Quecksilberausscheidung im Harn und Kot festgestellt, und zwar um 16,48% bei den Tieren, denen Unitiol in den Tagen 2–6 und um 12,87% bei denjenigen, denen Unitiol in den Tagen 8–14 nach Einführung des Quecksilbers verabreicht wurde. Bei den beiden Tiergruppen wurden Quecksilberablagerungen in den Nieren auf ca. 3–4 % herabgesetzt im Vergleich mit der Tiergruppe, an die kein Unitiol verabreicht wurde, wobei die Quecksilbermenge in den Nieren ca. 20% der eingefiihrten Dose betrug.     

Angiogenesis and cardioprotection after TNFalpha-inducer-Tolpa Peat Preparation treatment in rat's hearts after experimental myocardial infarction in vivo

The aim of the presented work was to evaluate whether short subcutaneous (s.c.) administration of TNFalpha-inducer-Tolpa Peat Preparation (TPP or TPP batch 0210) modulates the process of ischemic remodeling and spontaneous angiogenesis after experimental myocardial infarction (MI) in rats in vivo. The results obtained using three complementary and correlative methods: histological studies, Proliferating Cell Nuclear Antigen (PCNA) reaction and Lymphocytes Induced Angiogenesis (LIA) test showed a clear pro-angiogenic and cardioprotective effect of TPP administration after experimental MI. TPP batch 0210 should be considered as an angiogenesis stimulating factor and consecutively as a cardioprotective preventing development of ischemic cardiomyopathy after MI in rats. It might possibly be used as an adjunct to conventional therapy of coronary artery disease, including late phase after myocardial infarction or ischemic cardiomyopathy.     

Inhibition of plant and mammalian diamine oxidase by substrate analogues

Imidazoles, aliphatic substrate analogues and the natural dipeptides, carnosine and anserine, were investigated as inhibitors of diamine oxidase from the pig kidney, human pregnancy plasma and pea seedlings. Imidazole, methyli-midazoles,N-acetylimidazole, histamine andN -methyl-histamine are relatively potent inhibitors of mammalian diamine oxidase showing no influence on plant enzymes. Anserine and carnosine are inhibitors of pig kidney and pea seedling enzymes.K ₁ values are 2 M and 10 M respectively. Investigated natural derivatives of putrescine and cadaverine have no influence on diamine oxidase of different origin.In conclusion, we present some evidence to suggest that mammalian diamine oxidase, despite a high reaction rate with putrescine, is better adapted to histamine oxidation, whereas for plant enzymes the diamines are preferred substrates.     

Use of plate osteosynthesis as an effective method in surgical treatment of congenital cranial anomalies

Operative treatment of developmental anomalies were presented in actual work carried out in Maxillo-Facial Clinic PMA in Szczecin. Mesiocclusion was the most frequent anomaly, followed by microgenia, joint ankylosis of tempora mandibular, next laterogenia and first and second branchial arch syndrome. The operative treatment was finished by placing and fixing the bones in new changed position. The obtained results indicated that this type of management is the best effective in correction of visceral cranium development anomalies, it assure quick maintenance of good oral hygiene as well as stabilisation and functioning jaws.     

The evolution of epidemiology and clinical characteristics of laryngeal and hypopharyngeal carcinoma in Poland from 1991 to 2001

On the base of retrospective analysis of 12,888 cases of carcinoma of larynx and hypopharynx, diagnosed in 19 ENT Departments in Poland from 1991 to 2001, the assessment of basic epidemiological data, including the localization of tumor and stage of local and clinical advancement of the disease at the time of diagnosis has been conducted. In analyzed period of 11 years the trends to change of the mentioned above parameters has been examined. The significant increase of female patients in this period was observed, with average proportion M:F = 8:1. The glottis localization of carcinoma dominated (47.6%), followed by supraglottis (40.8%) and pyriform fossa (7.8%), with significant increase of pyriform fossa tumors in the analyzed period of 11 years. In the majority of cases the carcinoma of larynx and hypopharynx was diagnosed in the advanced stage (T3 + T4) of local disease, with the highest percentage in localization within the pyriform fossa (81.0%), and the lowest percentage in glottis tumors (45.6%). The regional lymph nodes metastases has been diagnosed in 46.7% of the analyzed group, with the highest percentage in tumors localized in pyriform fossa (82.9%), and the lowest percentage in tumors of glottis localization (33.1%). In the 11 years time the significant drop down of N0 cases and tendency to increase of N2 and N3 in the supraglottis localization of tumor. The distant metastases in the analyzed group at the time of diagnosis has been registered in 2.0%, with the highest percentage in posterior pharyngeal wall (7.6%) and pyriform fossa (7.4%). The authors postulate the renewal of prospective study on epidemiology, clinical characteristics and treatment results of larynx and hypopharynx carcinoma in Poland.     

Simultaneous Ionic and Electronic Conductivity in Polymeric Materials

Summary: Pyrrole and 3‐(3,6‐dioxaheptyl)pyrrole were polymerized in the presence of an oligo(oxyethylene monomethylether) sulfonate (DP_PEO = 17) with ammonium peroxydisulfate as an oxidizing agent and dry chloroform as a solvent. The oligo(oxyethylene) sulfonate acted as a phase transfer agent and also became incorporated into the polymer as a counter ion. The polymers were obtained as gel‐like particles that could be easily dispersed in organic solvent and could be doped with lithium triflate to enhance the ionic conductivity of the electronically conductive materials. DSC and X‐ray measurements showed a glass transition (T_g) in all samples at around ?40 °C which is related to the dynamics of the EO segments. In addition, the system composed of polypyrrole with oligo(oxyethylene) sulfonate counter ions showed partial crystallinity and a melting transition at 34 °C. Conductivity was explored by impedance spectroscopy with regard to frequency and temperature dependence. The data could be evaluated in the coordinates of an Arrhenius plot as the sum of two Arrhenius functions in all cases. Although one Arrhenius‐type function was sufficient to describe the temperature profile of the conductivity below T_g, a second term needed to be added at T > T_g. The absolute values of conductivity were also dependent on the level of doping with lithium triflate. These phenomena were interpreted as a contribution of ionic conductivity by the lithium ions to the electronic conductivity, which comes about from the oxidation state of the polypyrrole backbone.

A good Li‐ion conducting polymer which also exhibits electronic conductivity.     

High-resolution whole-organ mapping with SNPs and its significance to early events of carcinogenesis

We attempted to identify deleted segments in two model tumor suppressor gene loci on chromosomes 13q14 and 17p13 that were associated with clonal expansion of in situ bladder preneoplasia using single nucleotide polymorphisms (SNPs)-based whole-organ histologic and genetic mapping. For mapping with SNPs, the sequence-based maps spanning approximately 27 and 5 Mb centered around RB1 and p53, respectively, were assembled. The integrated gene and SNP maps of the regions were used to select 661 and 960 SNPs, which were genotyped by pyrosequencing. Genotyping of SNPs was performed on DNA samples corresponding to histologic maps of the entire bladder mucosa in human cystectomy specimens with invasive urothelial carcinoma. By using this approach, we have identified deleted regions associated with clonal expansion of intraurothelial neoplasia; which ranged from 0.001 to 4.3 Mb (average 0.67 Mb) and formed clusters of discontinuous deleted segments. The high resolution of such maps is a prerequisite for future positional targeting of genes involved in early phases of bladder neoplasia. This approach also permits analysis of the overall genomic landscape of the involved region and discloses that a unique composition of noncoding DNA characterized by a high concentration of repetitive sequences may predispose to deletions.