Cryptococcal lymphadenitis in a dog.

We describe a case of canine crytococcosis, the clinical symptoms were: feverish syndrome, vomiting and diarrhoeas and bilateral lymphadenitis in superficial lymph nodes. Microbiology and histopathology study of popliteal lymph node biopsy demonstrated the presence of round yeasts of some 3 microm of diameter, which we identified as Cryptococcus neoformans. Thirty months after suspending the medication the dog returned to the surgery; the dog was very thin and it had nervous symptoms; the owners decided it upon euthanasia. After carrying out the necropsy we take samples for their microbiology and histopathology study, both techniques detect to C. neoformans in marrow, CNS tissue and lymph nodes.     

RNA cleaving '10-23' DNAzymes with enhanced stability and activity

‘10-23’ DNAzymes can be used to cleave any target RNA in a sequence-specific manner. For applications in vivo, they have to be stabilised against nucleolytic attack by the introduction of modified nucleotides without obstructing cleavage activity. In this study, we optimise the design of a DNAzyme targeting the 5′-non-translated region of the human rhinovirus 14, a common cold virus, with regard to its kinetic properties and its stability against nucleases. We compare a large number of DNAzymes against the same target site that are stabilised by the use of a 3′-3′-inverted thymidine, phosphorothioate linkages, 2′-O-methyl RNA and locked nucleic acids, respectively. Both cleavage activity and nuclease stability were significantly enhanced by optimisation of arm length and content of modified nucleotides. Furthermore, we introduced modified nucleotides into the catalytic core to enhance stability against endonucleolytic degradation without abolishing catalytic activity. Our findings enabled us to establish a design for DNAzymes containing nucleotide modifications both in the binding arms and in the catalytic core, yielding a species with up to 10-fold enhanced activity and significantly elevated stability against nucleolytic cleavage. When transferring the design to a DNAzyme against a different target, only a slight modification was necessary to retain activity.     

Interrelations between monoaminergic afferents and corticotropin-releasing factor-immunoreactive neurons in the rat central amygdaloid nucleus: ultrastructural evidence for dopaminergic control of amygdaloid stress systems

Ample evidence implicates corticotropin-releasing factor (CRF)-producing neurons of the central amygdaloid nucleus (CeA) in vegetative, endocrine, and behavioral responses to stress and anxiety in laboratory rats. Monoaminergic systems are involved in modulating these responses. In the present paper, interrelations between CRF-immunoreactive (ir) neurons, and noradrenergic, serotonergic, and dopaminergic afferents were studied using single and double immunolabeling for light and electron microscopy in the rat CeA. Dopaminergic axons formed dense plexus in the CeA overlapping with the localization of CRF-ir neurons, and their terminals formed frequent associations with CRF-ir somata. Contacts of serotonergic axons on CRF-ir neurons were few, and contacts of noradrenergic axons were the exception. Ultrastructurally, symmetric synapses of dopaminergic terminals on CRF-ir somata and dendrites were found. More than 83% of CRF-ir somata were contacted in single ultrathin sections. About half of these possessed two or more contacts. Of non-ir somata, 37% were contacted by dopaminergic terminals, and only 13% of these had two or more contacts. Correlative in situ hybridization indicated that CeA CRF-ir neurons may express receptor subtype dopamine receptor subtype 2. In conclusion, dopaminergic afferents appear to specifically target CeA CRF neurons. They are thus in a position to exert significant influence on the rat amygdaloid CRF stress system.     

Identification of the three genotypic classes for soybean lipoxygenases 1 and 3 based on enzymatic activity

A non-destructive micro-test which allow the identification of homozygous and heterozygous soybean seeds for lipoxygenases (LOX) 1 and 3 has been developed based on determination of their enzymatic activities. Identification of heterozygous seeds will be extremely important in backcross breeding programme to expedite the creation of new soybean lines lacking seed lipoxygenases because no selfings are necessary during the odd numbered generations.     

3-(4-Aminobutyn-1-yl)pyridines: binding at alpha 4 beta 2 nicotinic cholinergic receptors

The binding of a series pyridylbutynylamines 6 was examined at alpha4beta2 nACh receptors. Structural modifications, comparing 6 with pyridyl ethers 2, did not consistently result in parallel effects on receptor affinity, suggesting possible differences in their modes of binding. Furthermore, the binding of amine 6a seemed to be accounted for by the newer vector pharmacophore models.     

New effects in polynucleotide release from cationic lipid carriers revealed by confocal imaging, fluorescence cross-correlation spectroscopy and single particle tracking

We report on new insights into the mechanisms of short single and double stranded oligonucleotide release from cationic lipid complexes (lipoplexes), used in gene therapy. Specifically, we modeled endosomal membranes using giant unilamellar vesicles and investigated the roles of various individual cellular phospholipids in interaction with lipoplexes. Our approach uses a combination of confocal imaging, fluorescence cross-correlation spectroscopy and single particle tracking, revealing several new aspects of the release: (a) phosphatidylserine and phosphatidylethanolamine are equally active in disassembling lipoplexes, while phosphatidylcholine and sphingomyelin are inert; (b) in contrast to earlier findings, phosphatidylethanolamine alone, in the absence of anionic phosphatidylserine triggers extensive release; (c) a double-stranded DNA structure remains well preserved after release; (d) lipoplexes exhibited preferential binding to transient lipid domains, which appear at the onset of lipoplex attachment to originally uniform membranes and vanish after initiation of polynucleotide release. The latter effect is likely related to phosphatidyleserine redistribution in membranes due to lipoplex binding. Real time tracking of single DOTAP/DOPE and DOTAP/DOPC lipoplexes showed that both particles remained compact and associated with membranes up to 1-2 min before fusion, indicating that a more complex mechanism, different from suggested earlier rapid fusion, promotes more efficient transfection by DOTAP/DOPE complexes.     

The effects of chronic aerobic and anaerobic exercises n lymphocyte subgroups

Exercise is the strongest stress to which the body is ever exposed. The body responds to this stress through a set of physiological changes in its metabolic, hormonal and immunological systems. In this study, responses of the immune system to the long-term aerobic and anaerobic exercises have been investigated. Twenty-four sedentary male university students and officers participated in this study. Subjects were divided into two groups, each consisting of twelve people. Group-1 (age: 25.67 +/- 3.79 years, height: 174.83 +/- 5.15 cm, body mass: 72.17 +/- 8.05 kg) and Group-2 (age: 24.83 +/- 2.89 years, height: 175.3 +/- 6.68 cm, body mass: 70.67 +/- 6.15 kg). After physical examinations of the two groups, resting ECG, respiratory function tests and metabolic tests with the use of the breath by breath method were completed, and anerobic heart rates at the threshold level were determined. The first group was subjected to exercise using Monark ergometry cycles at a heart rate 10% below the threshold level for 8 weeks, 3 days a week, 30 min a day. The second group exercised at a heart rate 10% above the threshold level for 8 weeks, 3 days a week, 20 min a day. Heart rates were checked with the Polar Test during exercises. Pre-exercise (Ep) venous blood samples were taken from each group before their 1st and 24th exercises. Hb (gr), Hct (%), erythrocyte (x10(6)/microl), leukocyte (x10(6)/microl), leukocyte subpopulations (neutrophil, lymphocyte, monocyte, eosinophil, basophil %) and thrombocyte (x10(6)/microl) values were determined. CD3, CD4, CD8, CD19 and CD56 values were determined by Flow Cytometry method using monoclonal antibodies. The chronic effects of exercise were examined through a comparison of Ep blood samples at the 1st exercise with Ep blood samples at the 24th exercise. While the increase in the total leukocyte number was significant (p<0.05) in the first group, increase in the second group was found to be non-significant. When percentiles of leukocyte subpopulations were taken into consideration, changes in the first and second group were found to be non-significant. When lymphocyte subgroups were examined; in the first group a decrease in CD3 and CD4 percentiles to 7% and 12%, respectively (p<0.05) and a 65% increase (p<0.01) in the CD56 value were observed. In the second group a decrease in CD3 and CD4 percentiles to 13% and 17%, respectively (p<0.05) and a 73% increase (p<0.01) in the CD56 value were observed. The Sample-t Test and The Wilcoxon Test were used for statistical analysis.     

The effect of L-NAME administrations after oral mucosal incision on wound NO level in rabbit

The aim of the current study was to comparatively investigate the effect of inhibition of nitric oxide (NO) production by N-nitro-L-arginine methyl ester (L-NAME), an isoform non-specific inhibitor of nitric oxide synthase (NOS), after oral mucosal incision on wound tissue NO levels. A standard incision was applied to the oral mucosa of rabbits. After oral mucosal incision, rabbits were divided into five groups as follows: (1) Untreated incisional group (control); (2) Titanium (Ti) implanted group; (3) Ti + Polyethylene glycol (PEG) 4000 implanted group; (4) Ti + PEG 4000 + L-NAME (2 × 10⁻⁴ M) implanted group and (5) i.p. L-NAME administrated group (10 mg/kg). At 5 days after oral incision operations, wound tissue strips and plasma were obtained from rabbits. Oral wound tissue and plasma nitric oxide, plasma thiobarbituric acid reactive substances (TBARS) and total sulfhydryl group (RSH) levels were investigated. Plasma TBARS and NOx levels decreased after i.p. L-NAME administration. Total RSH group levels were not changed in all groups (p>0.05). This means that L-NAME inhibits the deteriorating effects of free radicals without affecting healing. L-NAME in PEG and titanium also has no effect on tissue and plasma NOx levels. These findings indicate that NO generation will not be affected both Ti and local nitric oxide synthase (NOS) inhibitor. (Mol Cell Biochem 278: 65–69, 2005)