Species distribution and <Emphasis Type="Italic">in vitro</Emphasis> antifungal susceptibility of oral yeast isolates from Tanzanian HIV-infected patients with primary and recurrent oropharyngeal candidiasis

Background  

In Tanzania, little is known on the species distribution and antifungal susceptibility profiles of yeast isolates from HIV-infected patients with primary and recurrent oropharyngeal candidiasis.     

<Emphasis Type="Italic">K-ras,BRCA1/2</Emphasis>, and <Emphasis Type="Italic">CHEK2</Emphasis> mutations and loss of heterozygosity at 9p, 17p,and 18q in sporadic adenocarcinoma of the pancreas

The diagnostic significance of molecular markers was assessed for the most common somatic aberrations at the K-ras, TP53, CDKN2A, and MADH4 loci, as well as less common mutations of BRCA1, BRCA2, and CHEK2, arising in preinvasive stages of sporadic adenocarcinoma of the pancreas. The study was performed on paired primary pancreatic adenocarcinoma and normal pancreatic tissue specimens obtained from 37 Russian patients. Surgical adenocarcinoma specimens were subjected to manual microdissection. Mutations of K-ras codon 12 were found in 24 tumor specimens (0.65), but not in normal pancreatic tissue specimens. Mutations of BRCA1 (185delAG, 300T > G, 4153delA, 4158A > G, 5382insC), BRCA2 (695insT, 6174delT), and CHEK2 (1100delC) were not found. The informativeness of allelic losses did not differ significantly among the three tumor suppressor loci and was 60% for TP53 (GDB186817) and CDKN2A (D9S974 + D9S162) and 65.7% for MADH4 (D18S363 + D18S474) (t = 0.48). The CDKN2A locus had the highest LOH frequency of 0.95. For TP53 and MADH4 the LOH frequency was 0.62 and 0.70, respectively. In 80% of adenocarcinomas, at least one locus was characterized with LOH. The overall informativeness of the combined data on K-ras mutations and loss of heterozygosity at 9p, 17p, and 18q was 85.7%. Only 9% of the tumors were characterized with microsatellite instability.     

Effect of a new potential psychotropic drug, 8-(trifluoromethyl)-1,2,3,4,5-benzopentathiepin-6-amine hydrochloride,on the expression of serotonin-related genes in the mouse brain

Investigation of molecular mechanisms underlying psychotropic drug action is the main aim of molecular psychopharmacology. Previously, a new synthetic varacin analog, 8-(trifluoromethyl)-1,2,3,4,5-benzopentathiepin-6-amine (TC-2153) was shown to produce anxiolytic and anticonvulsant effects in mice. This study investigated the effects of chronic TC-2153 administration on the expression of some serotonin-related genes in the mouse brain. The drug was administered (10 mg/kg, per os, 16 days) to adult male mice of the ASC (Antidepressant Sensitive Catalepsy) strain characterized by altered behavior and hereditary impairment of the brain serotonin system. Expression of genes encoding tryptophan hydroxylase 2 (TPH2), the key enzyme of serotonin synthesis, monoamine oxydase A (MAOA), the major serotonin-degrading enzyme, 5-HT transporter (SERT), and 5-HT_1A receptor was studied using quantitative RT-PCR. TC-2153 significantly reduced the 5-HT_1A receptor and MAOA mRNA levels in the midbrain, but did not have any effect on the expression of these genes in the frontal cortex and the hippocampus. The drug did not affect the expression of TPH2 and SERT in the midbrain. The results indicate that the brain 5-HT system is involved in the molecular basis of TC-2153 action.     

Three dimensional typological studies using scanning electron microscopy for characterization of Termitomyces pellets obtained from submerged growth conditions

There are gaps in existing understanding of fungal pellet growth dynamics. We used scanning electron microscopy (SEM) for morphological characterization of the biomass organization of Termitomyces pellets for seven species: T. microcarpus (TMI1), T. albuminosus (TAL1, TAL2), T. striatus (TSTR), T. aurantiacus (TAUR), T. heimii (THE1, THE2), T. globulus (TGLO) and T. clypeatus (TCL1, TCL2, TCL3, TCL4, TCL5). We assessed the utility of SEM for morphological and structural characterization of Termitomyces spp. in three dimensional (3D) pellet form to identify ideal pellet morphology for industrial use. Typological classification of Termitomyces species was based on furrows, isotropy, total motifs and fractal dimensions. The pellets formed were entangled and exhibited highly compacted mycelial mass with microheterogeneity and microporosity. The mean density of furrows of Termitomyces species was between 10,000 and 11,300 cm/cm², percentage isotropy was 30?80 and total motifs varied from 300 to 2500. TGLO exhibited the highest furrow mean density, 11243 cm/cm², which indicated a compact, cerebroid structure with complex ridges and furrows, whereas TAL2 exhibited the lowest furrow density. TMI1a exhibited a high percentage isotropic value, 74.6, TSTR exhibited the lowest, 30.9. Total motif number also was used as a typological classification parameter. Fractal values were 2.64?2.78 for various submerged conditions of Termitomyces species. TAL1 exhibited the highest fractal dimension and TAL2 the lowest, which indicates the complexity of branching patterns. Three-dimensional SEM image analysis can provide insight into pellet micromorphology and is a powerful tool for exploring topographical details of pellets.     

Adverse effects of tempol on hidrosaline balance in rats with acute sodium overload

We studied the effects of tempol, an oxygen radical scavenger, on hydrosaline balance in rats with acute sodium overload. Male rats with free access to water were injected with isotonic (control group) or hypertonic saline solution (0.80 mol/l NaCl) either alone (Na group) or with tempol (Na-T group). Hydrosaline balance was determined during a 90 min experimental period. Protein expressions of aquaporin 1 (AQP1), aquaporin 2 (AQP2), angiotensin II (Ang II) and endothelial nitric oxide synthase (eNOS) were measured in renal tissue. Water intake, creatinine clearance, diuresis and natriuresis increased in the Na group. Under conditions of sodium overload, tempol increased plasma sodium and protein levels and increased diuresis, natriuresis and sodium excretion. Tempol also decreased water intake without affecting creatinine clearance. AQP1 and eNOS were increased and Ang II decreased in the renal cortex of the Na group, whereas AQP2 was increased in the renal medulla. Nonglycosylated AQP1 and eNOS were increased further in the renal cortex of the Na-T group, whereas AQP2 was decreased in the renal medulla and was localized mainly in the cell membrane. Moreover, p47-phox immunostaining was increased in the hypothalamus of Na group, and this increase was prevented by tempol. Our findings suggest that tempol causes hypernatremia after acute sodium overload by inhibiting the thirst mechanism and facilitating diuresis, despite increasing renal eNOS expression and natriuresis.     

Fasting protects against the side effects of irinotecan but preserves its anti-tumor effect in Apc15lox mutant mice

Irinotecan is a widely used topoisomerase-I-inhibitor with a very narrow therapeutic window because of its severe toxicity. In the current study we have examined the effects of fasting prior to irinotecan treatment on toxicity and anti-tumor activity. FabplCre;Apc^15lox/+ mice, which spontaneously develop intestinal tumors, of 27 weeks of age were randomized into 3-day fasted and ad libitum fed groups, followed by treatment with a flat-fixed high dose of irinotecan or vehicle. Side-effects were recorded until 11 days after the start of the experiment. Tumor size, and markers for cell-cycle activity, proliferation, angiogenesis, and senescence were measured. Fasted mice were protected against the side-effects of irinotecan treatment. Ad libitum fed mice developed visible signs of discomfort including weight loss, lower activity, ruffled coat, hunched-back posture, diarrhea, and leukopenia. Irinotecan reduced tumor size in fasted and ad libitum fed groups similarly compared to untreated controls (2.4 ± 0.67 mm and 2.4 ± 0.82 mm versus 3.0 ± 1.05 mm and 2.8 ± 1.08 mm respectively, P < 0.001). Immunohistochemical analysis showed reduced proliferation, a reduced number of vascular endothelial cells, and increased levels of senescence in tumors of both irinotecan treated groups. In conclusion, 3 days of fasting protects against the toxic side-effects of irinotecan in a clinically relevant mouse model of spontaneously developing colorectal cancer without affecting its anti-tumor activity. These results support fasting as a powerful way to improve treatment of colorectal carcinoma patients.     

Polymorphism and structural features of two noncoding regions of the liver fluke <Emphasis Type="Italic">Fasciola hepatica</Emphasis> (Plathelminthes: Trematoda) mitochondrial genome

Structural characteristics and polymorphism of the long (LNR) and short (SNR) mitochondrial noncoding regions were studied in the liver fluke Fasciola hepatica. Flukes were sampled from several populations of Russia and Belarus. LNR amplification yielded a set of nine fragments, neighboring ones differing in length by one tandem repeat (85 bp), published for Australian flukes. The LNR amplification products of different lengths were cloned and sequenced. A comparison of the LNR sequences of Australian and Belarussian flukes revealed three nucleotide substitutions and one point heteroplasmy in the first positions of the imperfect repeat and four adjacent perfect repeats. The positions of the three mutations coincided in the perfect and imperfect repeats. The frequency of mutations was 4.0–4.7 %, while the frequency of heteroplasmic sites varied from 0.1 to 1.2%. It was shown that the mutations and the heteroplasmy of one site could change the structure and stability of the putative secondary structures of the perfect and imperfect repeats. SNR amplification in F. hepatica from several populations yielded fragments that differed from the published SNR sequence of Australian F. hepatica by one transversion (T → G in position 21). Both noncoding regions had several conserved and potential regulatory sequences. The possible causes of heteroplasmy and a concerted origin of substitutions in different repeats are discussed.     

Mutation analysis of <Emphasis Type="Italic">K-ras</Emphasis> protooncogene in colorectal adenocarcinomas and polyps in Russian patients

To estimate diagnostic value of K-ras mutations during cancer risk group formation, they were studied in the samples of sporadic carcinomas (n = 58), benign (n = 33), and malignant (n = 13) polyps of large intestine obtained during surgery or polypectomy. Using PCR analysis, restriction analysis, SSCP analysis and automated sequencing, eight various point mutations were revealed. Six of them were located in codon 12 and two, in codon 13 of the K-ras gene. Mutation frequency in carcinomas, benign and malignant polyps was 43, 49, and 69%, respectively. In the normal tissue samples of colorectum, no changes in codons 12 and 13 in the K-ras gene were observed. Mutations in the groups of Russian patients examined partially overlapped. In patients with colorectal carcinoma the mutation frequency in the K-ras gene was not associated with disease onset age, location, and the extent of tumor differentiation while it was associated with the stage of tumor process. In polyps, the maximum mutation frequency was revealed among patients over 70 years of age as well as in the adenomas of villous histology and large size (≥1cm). No correlation between the K-ras mutation frequency and the extent of polyp dysplasia was observed.     

MIDAS: software for analysis and visualisation of interallelic disequilibrium between multiallelic markers

Background  

Various software tools are available for the display of pairwise linkage disequilibrium across multiple single nucleotide polymorphisms. The HapMap project also presents these graphics within their website. However, these approaches are limited in their use of data from multiallelic markers and provide limited information in a graphical form.     

Synthesis of polymeric neoglycoconjugates based onN-substituted polyacrylamides

Several types of polymeric glycoconjugates,N-substituted polyacrylamides, have been synthesized by the reaction of activated polymers with -aminoalkylglycosides: (i) (carbohydrate-spacer) _n -polyacrylamide, pseudopolysaccharides; (ii) (carbohydrate-spacer) _n -phosphatidylethanolamine _m -polyacrylamide, neoglycolipids, derivatives of phosphatidylethanolamine; (iii) (carbohydrate-spacer) _n -biotin _m -polyacrylamide, biotinylated probes; (iv) (carbohydrate-spacer) _n -polyacrylamide-(macroporous glass), affinity sorbents based on macroporous glass, covalently coated with polyacrylamide. An almost quantitative yield in the conjugation reaction makes it possible to insert in the conjugate a predetermined quantity of the ligand(s).Pseudopolysaccharides proved to be a suitable form of antigen for activation of polystyrene and poly(vinyl chloride) plates (ELISA) and nitrocellulose membranes (dot blot), being advantageous over traditional neoglycoproteins. Polyvalent glycolipids insert well in biological membranes: their physical properties, particularly solubility, can be changed in a desired direction. Biotinylated derivatives were used as probes for detection and analysis of lectins.Abbreviations sp spacer arm - PAA polyacrylamide - PE phosphatidylethanolamine - Biot biotin - MPGlass macroporous glass 200 Å - ELISA enzyme-linked immunosorbent assay - HPLC high-performance liquid chromatography - AIBN azodiisobutyronitrile - BSA bovine serum albumin - DMG 3,6-di-O-methyl-d-glucose - TLC thin-layer chromatography - DMF dimethylformamide - DMSO dimethylsulfoxide - RI refractive index - PBS phosphate buffered saline (0.14m NaCl, 0.01m sodium phosphate, pH 7.3)