Internal amplification control of PCR for the Glu1-Dx5 allele in wheat.

One of the limiting factors in using dominant markers is the unique amplification of the target fragment. Therefore, failures in polymerase chain reaction (PCR) or non-amplifications can be interpreted as an absence of the allele. The possibility of false negatives implies in reduced efficiency in the selection process in genetic breeding programs besides the loss of valuable genetic material. Thus, this study aimed to evaluate the viability of a microsatellite marker as an internal amplification control with a dominant marker for the wheat Glu1-Dx5 gene. A population of 77 wheat cultivars/breeding lines was analyzed. Fourteen microsatellite markers were analyzed in silico regarding the formation of dimers and clamps. The biplex reaction conditions were optimized, and the Xbarc117 marker was selected as the internal amplification control with a Glu1-Dx5 marker in wheat. It was concluded that the Xbarc117 microsatellite marker was effective in the simultaneous amplification with a dominant Glu1-Dx5 marker, making biplex PCR viable in wheat for the studied markers.

Genetic polymorphisms and retinal vein occlusion in an Italian population.

In this study, we assessed the prevalence of polymorphisms in genes involved in hyperhomocysteinemia or hemostasis to shed light on their role, if any, in retinal vein occlusion (RVO). We recruited 37 Italian patients (17 men and 20 women) with a diagnosis of central or branch RVO based on fundus examination and retinal fluorescein angiography, as well as 45 healthy controls. Risk factors and family history of RVO of all subjects were recorded. The distributions of polymorphisms in patients and controls were evaluated using the χ(2) test and OR. We confirmed an increased risk in subjects with dyslipidemia (high density lipoprotein <59 mg/dL: 17.8% of controls, 43.2% of patients, P = 0.0002; low density lipoprotein >130 mg/dL: 26.7% controls, 54.1% patients, P = 0.0002), arterial hypertension (60% controls, 75.7% patients, P = 0.023), and high body mass index (28.9% controls, 70.3% patients, P < 0.0001, and excluded involvement of the selected polymorphisms in RVO. Overall, the tested polymorphisms did not appear to be useful for assessing predisposition or for the diagnosis and prognosis of RVO.

Sobrepoblación aanina y felina: tendencias y nuevas perspectivas / Canine and feliee overpopulation: trends and new perspectives

A través de una revisión sistemática de la literatura, publicada entre 1973 y 2009, se consultaron las siguientes bases de datos, a saber: ScienceDirect, Ebsco, Springerlink y Medline, para la búsqueda de la información. Como palabras clave se utilizaron: perros de libre ambulación, vínculo animal-humano, población de mascotas, sobrepoblación y control de población. Además se consultó el banco de publicaciones de la Sociedad Mundial para la Protección Animal (WSPA) y la Organización Internacional de Epizootias (OIE). El objetivo de esta revisión fue presentar una posición crítica sobre la problemática de la sobrepoblación de mascotas, su percepción cultural y la relación hombre-animal. Asimismo, los fracasos asociados con esta relación, estableciendo posibles soluciones sin desconocer nuestro marco cultural. Se obtuvo un total de dieciséis referencias, a las que se aplicó los criterios de inclusión y exclusión; los artículos que cumplieron estos criterios son los que constituyeron la unidad de análisis de esta revisión. A medida que aumente nuestro conocimiento acerca de la tenencia responsable de las mascotas, mejorarán nuestros vínculos afectivos con estas. Solo a través de la educación sanitaria se puede adquirir el conocimiento necesario para evitar fracasos con respecto a una tenencia adecuada, de allí la responsabilidad y el papel fundamental que ejerce el médico veterinario en la comunidad.

Through a systematic review of literature comprising publications between 1973 and 2009, the following databases were consulted: Science Direct, EBSCO, Springer Link and MEDLINE. Key words used included stray dogs, animal-human bond, pet population, overpopulation and population control. The databases of the World Society for the Protection of Animals (WSPA) and the International Organization of Epizootics (OIE) were also consulted. The objective of the present review was to present a critical position about the pet overpopulation problem, cultural perception and relationships between human and animals, and failures associated with this relationship with the purpose of establishing possible solutions to the problem taking into consideration cultural issues. A total of 16 references were found, which, we applied the inclusion and exclusion criteria. Articles that met these criteria are those that constitute the unit of analysis of this review. As the knowledge about responsible ownership of pets is increased, the bond with them is enhanced. Only through health education, it is possible to acquire the necessary knowledge needed to avoid failures with respect to proper keeping. In this respect the veterinarian plays a pivotal role with his community.

Prevalence of Toxoplasma gondii in dogs from Colombia, South America and genetic characterization of T. gondii isolates.

The prevalence of Toxoplasma gondii in 309 unwanted dogs from Bogotá, Colombia, South America was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT) and found in 52 (16.8%) of 309 dogs with titers of 1:20 in 20, 1:40 in six, 1:80 in 17, 1:160 in three, 1:320 in three, 1:1280 or higher in three. Some organs obtained after necropsy of dogs (hearts, tongues and brains, either separately or pooled) were used in bioassays carried out in mice (37 samples, of which 20 were assayed with separate organs and 17 were assayed with pooled organs), cats (pooled organs from six) and pooled organs of two dogs both in mice and cat. Mice receiving dog tissues were examined for T. gondii infection. Feces of cats that received dog tissues were examined for oocyst shedding. In total, T. gondii was isolated from tissues of 20 dogs (16 by bioassays in mice, 3 by bioassay in cats and 1 by bioassay in mice and cat). All infected mice from 7 of 17 isolates bioassayed in this host died of toxoplasmosis during primary infection. Only 10 of the 20 dogs whose tissues were bioassayed separately induced infections in mice. Interestingly, dog organs varied in their capacity to induce T. gondii infection in mice, hearts and tongues producing more positive results than the brain. The 20 T. gondii isolates obtained from seropositive dogs were PCR-RFLP genotyped using polymorphisms at 10 nuclear markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, a new SAG2 and an apicoplast marker Apico. Ten genotypes were revealed. These genotypes are different from the three predominant Types I, II and III lineages that are widely spread in North America and Europe. A new allele denoted u-3 at PK1 locus was identified in three isolates. This result supports previous findings that T. gondii population is highly diverse in Colombia.

Prevalence of Toxoplasma gondii in cats from Colombia, South America and genetic characterization of T. gondii isolates.

Cats are important in the epidemiology of Toxoplasma gondii infection because they are the only hosts that can excrete the environmentally-resistant oocysts. In the present study, prevalence of T. gondii was determined in serum, feces, and tissues of 170 unwanted cats from Colombia, South America. Antibodies to T. gondii were assayed by the modified agglutination test and found in 77 of 170 (45.2%) cats with titers of <1:5 in 93, 1:5 in eight, 1:10 in 17, 1:20 in 10, 1:40 in seven, 1:80 in four, 1:160 in eight, 1:320 in six, and 1:640 or higher in 17 cats. T. gondii oocysts were not found in feces of any cat as ascertained by bioassay in mice. Tissues (brain, heart, tongue) of 116 cats were bioassayed in mice or cats. T. gondii was isolated from tissues of 15 of the 42 cats with titers of 1:40 or higher and not from any of the 90 cats titers of 1:20 or lower. Of the 29 cats whose tissues were bioassayed individually, T. gondii was isolated from the tongues of nine, hearts of eight, and brains of five. Mice inoculated with tissues of 12 of 15 infected cats died of toxoplasmosis; with nine T. gondii isolates all infected mice died. Overall, 65 of 92 (70%) of T. gondii-infected mice died of toxoplasmosis. Genotyping of these 15 isolates using polymorphisms at the SAG1, SAG2, SAG3, BTUB, and GRA6 loci revealed that three isolates (TgCtCo1, 2, and 7) had Type I alleles and one isolate (TgCtCo8) had Type II allele at all five loci. Eleven isolates contained the combination of Type I and III alleles and were divided into three genotypes, with TgCtCo3,5,6,9,12,13 and 15 had alleles I, I, III, I and III, TgCtCo4,10,11 had alleles I, III, III, I and I, and TgCtCo14 had alleles I, III, III, III, and III, at loci SAG1, SAG2, SAG3, BTUB and GRA6, respectively. All infected mice from each group had identical genotype except one mouse infected with TgCtCo5 had a Type III allele at locus BTUB and a unique allele (u-1) at locus SAG1 indicating mixed infection for TgCtCo5, whereas the rest seven mice had a Type I alleles at both loci.

Olanzapine acute administration in schizophrenic patients increases delta sleep and sleep efficiency.

BACKGROUND: A delta sleep deficit has been observed in schizophrenic patients. Olanzapine is a novel atypical antipsychotic agent with affinity at dopaminergic, serotonergic, muscarinic, adrenergic and histaminergic binding sites. The present study was designed to analyze a sleep promoting effect reported for olanzapine. METHODS: Twenty schizophrenic patients (DSM-IV) were studied, who were drug free and inpatients. Patients slept for 5 consecutive nights in the sleep unit as follows: one acclimatization night; two baseline nights (the first for sleep disorder screenings); and two olanzapine nights (10 mg olanzapine, one hour before sleep onset). RESULTS: Sleep continuity variables and total sleep time showed an overall improvement with olanzapine. Waking time was reduced since the first night of olanzapine administration. The main sleep architecture changes were: reduction in sleep stage 1, while sleep stage 2 and delta were significantly enhanced. Rapid eye movement density was also increased by the second olanzapine night. CONCLUSIONS: Total sleep improvement was due to the increase in sleep stages 2 and delta sleep. This may be related to serotonergic antagonistic properties of olanzapine. Olanzapine seems to have a sleep promoting effect in schizophrenic patients.

Photochemical and pharmacokinetic properties of selected flavins.

Some photochemical and pharmacokinetic properties of riboflavin, lumiflavin and the 2',3',4',5' tetraacetyl, tetrapropionyl, tetrabutiryl and tetrapalmitoyl esters of riboflavin have been studied. The esters are considerably more photostable than riboflavin but less so than lumiflavin, and appear to be photosensitizers with a behaviour similar to that of riboflavin, promoting photoreactions of biological targets even in the absence of molecular oxygen. The various flavins display important differences in their pharmacokinetic behaviour. Riboflavin, lumiflavin and the short-chain esters (the acetyl and propionyl esters) are rapidly cleared from serum, and recovered in comparable amounts from the liver and kidneys. These results are in agreement with their hydrophilic or moderately hydrophobic character. In contrast, the longer-chain butiryl and palmitoyl esters exhibit a prolonged retention in serum and undergo a significantly larger accumulation in the liver as compared with the kidneys; they are also found in the spleen. In all cases the tissue uptake of these esters becomes appreciable only after 24 h. These results are coherent with the highly hydrophobic character of these esters, which induce a slow release from serum lipoproteins and have a preferential clearance via the bile-gut pathway, showing affinity for the components of the reticuloendothelial system. These long-chain riboflavin esters will probably have a greater and more persistent risk of photoinduced hepatotoxicity than riboflavin, lumiflavin and the short-chain esters.

Development of tolerance after repeated administration of a selective muscarinic M1 antagonist biperiden in healthy human volunteers.

The muscarinic antagonist biperiden produces a dose-dependent inhibition of (REM) sleep on acute administration. The present study addressed the possibility of pharmacological tolerance after repeated biperiden administration. Six healthy volunteers were studied under sleep laboratory conditions in the following situations: one acclimatization, night, two baseline (that were averaged), 4 nights of biperiden administration, and 4 nights of placebo recovery administration. Six milligrams of biperiden and placebo were administered in identical capsules. Volunteers and technicians were blind to the order of the administration of the capsules. REM sleep time was reduced during the first and the second night, but was not significantly different in comparison with baseline by the third night. During placebo recovery nights, REM sleep time was not different from baseline. REM sleep latency was increased during the first and second nights of biperiden administration, but tolerance to this effect was observed by the third night. On placebo nights a dramatic shortening of REM latency was observed. The present findings support the hypothesis that anticholinergic drugs, even a selective M1 antagonist such as biperiden, induce tolerance soon after administration. A similar effect has been reported with scopolamine, a nonselective muscarinic antagonist, but the main difference is that biperiden withdrawal was not followed by an REM sleep rebound. The observed effect on REM sleep latency during placebo administration may be related to a supersensitivity to muscarinic M-1 receptors that trigger the first REM sleep period. Because short REM latency has been the main finding in the sleep of depressed patients, some implications of the present findings are discussed.

Rapid eye movement (REM) sleep increases by auditory stimulation reverted with biperiden administration in normal volunteers.

Auditory stimulation during REM sleep increases REM sleep time. The purpose of the present work was to determine if the selective muscarinic M-1 antagonist biperiden could modify the effect of the auditory stimulation on REM sleep. Twelve healthy volunteers were divided into placebo and biperiden groups. All the volunteers were studied under sleep laboratory conditions as follows: one acclimatization night, one baseline night, four nights with auditory stimulation either with placebo or biperiden, and two follow-up nights. Biperiden (4 mg) or placebo in identical capsules was administered 1 hour before beginning the sleep recordings. REM sleep time and REM density in the placebo group were increased relative to baseline by auditory stimulation. Biperiden blocked the REM time increase over the three treatment nights and suppressed the REM density increase over all four treatment nights. Biperiden also increased the latency compared to the placebo group. The present findings suggest that M-1 mechanisms are related to REM sleep regulation.