An efficient protocol for genomic DNA extraction from formalin-fixed paraffin-embedded tissues

Formalin-fixed paraffin-embedded tissues (FFPET) represent the largest source of archival biological material available for genomic studies. In this work we present an advanced protocol for extraction of high quality DNA from FFPET that can be applied in several molecular studies. Although cat mammary tumours (CMT) are the third most frequent tumour in cats the recovery of significant number of samples for molecular studies are in some way restricted to FFPET samples. We were able to obtain high quality DNA from FFPET of thirty six CMT that were subjected to pre-fixation and fixation processes routinely used in the veterinary hospitals. The quality of DNA obtained was tested by PCR amplification using six sets of primers that amplify single-copy fragments. The DNA fragments obtained were further sequenced. This protocol was able to provide FFPET gDNA that can be amplified and sequenced for larger fragments up to 1182bp.     

Use of land-races in Czechoslovak wheat breeding

Summary Four groups of Czechoslovak land-races (Czech alternate, Czech red, Moravian white, awnless, South Moravian and Slovak awned wheats) are characterized on the base of wheat collection studies in the Research Institute for Crop Production, Praha-Ruzyn, and their use in the breeding in SSR. A review of Czechoslovak cultivars of winter and spring wheats registered in the years 1921–1980 (table 1 and 2) and the registration period of the most important older Czechoslovak cultivars (table 3) is presented.Of the total number of 284 registered Czechoslovak cultivars in the years 1921–1980 93 cultivars were developed by selection from land-races; 70% of 87 cultivars originated from crosses had a land-race or its derivative as one of its parents. From land-races derived cultivars Chlumecká 12, eská Pesívka, Slovenská 777, Dobrovická 10 displayed the longest duration in the registration.

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Die Verwendung von Landsorten in der tschechoslowakischen Weizenzüchtung

Zusammenfassung Vier Gruppen von tschechoslowakischen Weizenlandsorten werden auf der Grundlage von Untersuchungen an der Weizenkollektion im Institut für Pflanzenproduktion, Prag-Ruzyn, und ihrer Nutzung in der Züchtung in der SSR beschrieben. Es sind (1) tschechische Wechselweizen, (2) tschechische rote, (3) mährische weiße, grannenlose und (4) südmährische und slowakische begrannte Weizen. Ein Überblick über die in den Jahren 1921–1980 registrierten tschechoslowakischen Winter- und Sommerweizen (Tab. 1 und 2) sowie die Zeitdauer der Registrierung der bedeutendsten älteren tschechoslowakischen Sorten (Tab. 3) schließt sich an.Von insgesamt 284 in den Jahren 1921–1980 registrierten tschechoslowakischen Sorten entstanden 93 durch Auslese aus Landsorten. Bei den 87 Sorten, die aus Kreuzungen hervorgingen, hatten 70% entweder eine Landsorte oder den Abkommen einer Landsorte unter den Eltern. Die aus Landsorten hervorgegangenen Sorten Chlumecká 12, eská Pesívka, Slovenská 777 und Dobrovická 10 waren am längsten registriert.

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- : (1) , (2) , (3) , (4) . , 1921–1980 . (. 1 2), ( ) (. 3). 284 , 1921–1980 ., 93 ë . 87- , ë , 70% . ( ) : 12, , 777 10.

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Paper not delivered during the symposium     

Effect of drying conditions and storage period on polyphenolic content, antioxidant capacity, and ascorbic acid of prunes

In this study the main chemical parameters, ascorbic acid and polyphenol content, and antioxidant activity of two varieties of prunes, dried by high-temperature (85 + 70 degrees C) and low-temperature (60 degrees C) procedures, were monitored during storage. Ascorbic acid content was higher in the prunes dried at 60 degrees C but significantly decreased in both varieties during storage. The different classes of polyphenols analyzed (cinnamates, anthocyanins, flavonols) showed different stabilities during storage. Neochlorogenic acid decreased only in the President variety, whereas chlorogenic acid increased in both varieties; anthocyanins, present only in the President prunes, disappeared in the first months of storage, and the flavonol content fell significantly in both cultivars during the year of the study. Drying temperature significantly affected the polyphenol content, with different effects according to the class of polyphenols. Antioxidant activity showed a significant increase at the end of the storage period and in the President variety was higher in the sample dried at the higher temperature.     

Sugar yield,nitrogen uptake by sugar beet and optimal nitrogen fertilization in relation to nitrogen soil analyses and several additional factors

To test the relative usefulness of different methods of chemical analysis for soil nitrogen fractions in the assessment of the fertilizer nitrogen needs of sugar beet, different doses of nitrogen were applied in field experiments laid out during the years 1985–1991. The chemical methods used were N mineral (NO _– ³ +NH ₊ ⁴ ) analyses on soil samples taken in late winter, and extraction with 0.01 M CaCl₂ from soil samples taken the preceding autumn and in late winter. The results of the chemical methods were evaluated in models using estimated optimum nitrogen fertilization, nitrogen present in beets or beets+leaves at leaf maximum and sugar yield as variables. In addition, parameters such as estimates of possible rooting depth and mineralization capacity of the soil were also included in the model. All models for estimating nitrogen fertilization need showed low R ² values. The two methods of soil chemical analysis yielded similar R ² values for nitrogen uptake in plots both with and without nitrogen fertilization. The N mineral method was least useful in predicting sugar yield. Addition of the covariables rooting depth and mineralization capacity appreciably improved the explanatory value of the models with 0.01 M CaCl₂, especially when the analytical results of soil samples taken in autumn were used. For the N mineral method the addition of covariates was found to have far less influence.     

Factors contributing to the development of aluminum resistance in the Madeiran maize germplasm

Forty genotypes representing maize genetic diversity from the Island of Madeira were screened for resistance to aluminum (Al) in nutrient solution. Seeds of maize were obtained from local farmers from field plots spread around the island in a range of altitudes from 99 to 1000 m above sea level. The content of ionic aluminum and the pH of soils cultivated with maize were determined. The mean pH value of all examined soils was 4.9, while the mean ionic Al content was 0.76 cmol kg^–1. Seventy‐two hours (h) exposure to 100 and 200 μM Al followed by a 48 h recovery period of 4‐day old seedlings revealed significant differences in Al resistance in the Madeiran maize germplasm. Root survival and regrowth exceeding 80 % of the seedlings were observed in 22 and 8 of the tested genotypes screened at 100 and 200 μM Al in nutrient solution. High Al resistance among Madeiran maize genotypes appeared not to be associated with the lower pH of soil, and did not correlate with the amount of soil Al³⁺ or the altitude at the site of collection. The high level of Al resistance of the maize cultivars indicates an initial genetic trait enhanced by open pollination of maize.     

Transient interface sharpening in miscible alloys

We observed that diffuse interfaces sharpen rather than broaden in completely miscible ideal binary systems. This is shown in situ during heat treatments at gradually increasing temperatures by scattering of synchrotron radiation in coherent Mo/V multilayers containing initially diffuse interfaces. This effect provides a useful tool for the improvement of interfaces and offers a way to fabricate better x-ray or neutron mirrors, microelectronic devices, or multilayers with giant magnetic resistance.     

Effects of rumen-protected oil supplementation on finishing grazing beef cattle

Tropical Animal Health and Production - The objective of this study was to evaluate the effects of rumen-protected oil (soybean and palm oil) in supplements for beef cattle during the fattening...     

Ambulatory esophageal pHmetry in healthy dogs with and without the influence of general anesthesia

This research aimed to determine the value of esophageal pH in awake and anesthetized dogs, to evaluate the esophageal pH value in awake dogs, in different body positions, as well as to study the occurrence of gastroesophageal reflux episodes in these positions. Thus, 40 healthy male and female adult dogs with mean body weight of 15.5 ± 4.6 kg were used. Esophageal pHmetry was conducted by inserting a catheter through the oropharynx in 30 dogs (stage 1) anesthetized with acepromazine, propofol and isoflurane, submitted to elective ovariosalpingohysterectomy. In addition, 8-h esophageal pHmetry was carried out transnasally in 10 awake dogs (stage 2), allowed to move and change body positions (lateral and sternal decubitus, and standing position), which were recorded. The mean esophageal pH value was lower (p < 0.01) in the anesthetized dogs (7.3 ± 0.82) than in the awake dogs (8.2 ± 0.3). Only four anesthetized dogs (13.33%) suffered reflux episodes. Reflux was not observed in the awake dogs and no esophageal pH differences were found between the body positions studied. Compared to the alert state, general anesthesia in dogs submitted to the previously mentioned anesthesia protocol causes esophageal pH reduction and predisposes to the occurrence of gastroesophageal reflux episodes. Transnasal pHmetry of 8 h in healthy awake dogs reveals that the esophageal pH value is alkaline and does not vary according to body position. In these animals, decubitus position is not a determining factor for reflux episodes to occur.     

Molecular characterization of Corynebacterium pseudotuberculosis isolates using ERIC-PCR

Caseous lymphadenitis is an infectious sheep and goats disease caused by Corynebacterium pseudotuberculosis and characterized by abscesses in superficial and visceral lymph nodes. C. pseudotuberculosis strains isolated from these hosts have been shown to be very difficult to type by the existing methods. The aim of this study is evaluating the potential of the Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) as a tool for molecular typing of C. pseudotuberculosis strains isolated in sheep. One hundred and twenty seven isolates of C. pseudotuberculosis were isolated from lesions suspected to have had caseous lymphadenitis collected from sheep at the slaughterhouse. Animals were from 24 flocks in 13 municipalities of the Minas Gerais State, Brazil. Species identification of the isolates was performed by routine biochemical tests and mPCR. Fingerprint was performed by RAPD using ERIC-1R, ERIC-2 and ERIC-1R+ERIC-2 primers. Seventeen different genotypes were generated by ERIC 1-PCR, 21 genotypes by ERIC 2-PCR and 21 genotypes by ERIC 1+2-PCR. Hunter-Gaston Discrimination Index (HGDI) found for ERIC 1, ERIC 2, ERIC 1+2 PCR were 0.69, 0.87, and 0.84, respectively. For most herds evaluated observed at most three different genotypes among isolates from animals of these property, in all ERIC-PCR assays. However a few flocks observed between four and nine genotypes per flock. The W Kendall value found for correlation among the three techniques of ERIC-PCR was 0.91 (P<5.0 x 10(-6)). The results show that ERIC-PCR has good discriminatory power and advantages over other DNA-based typing methods, making it a useful tool to discriminate C. pseudotuberculosis isolates.     

Outbreak of Mycobacterium avium subsp. avium infection in one flock of domestic pigeons

An outbreak of Mycobacterium avium subsp. avium infection was diagnosed in one breed of domestic pigeons (Columba livia f. domestica) in the Czech Republic. Nodular granulomatous lesions were found in 42 (9.7%) pigeons of the 435 examined; histopathologic examination of livers with gross lesions of mycobacteriosis from 15 randomly selected pigeons revealed granulomatous inflammation typical for avian mycobacteriosis in all samples. Direct Ziehl-Neelsen (ZN) microscopy and conventional culture were performed for a total of 117 liver samples (42 pigeons with nodular lesions, 55 randomly selected pigeons without nodular lesions, and 20 randomly selected squabs). Acid-fast bacilli were observed in 19 (16.2%), and conventional culture yielded growth of M. a. avium in 40 (34.2%) liver samples. A triplex quantitative real-time PCR assay based on the IS901 detection system was performed successfully in 115 liver samples and revealed M. a. avium in 63 (54.8%) of them. Mycobacterium a. avium was also detected in two squabs. Eight domestic rabbits (Oryctolagus cuniculus f. domestica) living in the breeding facility were also examined. Pyogranulomatous lesions were only found in one adult male rabbit. At necropsy, both direct ZN microscopy and culture gave negative results for mycobacteria in all examined rabbit tissues. Mycobacterium a. avium was diagnosed in a liver sample of one juvenile rabbit using triplex qPCR, suggesting that M. a. avium infection can occur as early as juvenile animals.