Imerslund-Grasbeck syndrome in a cross-breed dog

Imerslund-Gräsbeck syndrome is an autosomal recessive disease reported only in certain pure-breed dogs. An 18-month-old, male neutered beagle cross-breed was presented for evaluation of severe lethargy, progressive weakness and anorexia. Main clinicopathological findings included low body condition score (2.5/9), severe muscle atrophy, several neurological abnormalities, mild normochromic, normocytic, non-regenerative anaemia, severe hypocobalaminemia and mild proteinuria. Extensive diagnostic tests ruled out most of differential diagnoses for the aforementioned clinicopathological abnormalities and genetic evaluation showed that the dog was heterozygous for two previously described mutations affecting the CUBN gene, the beagle and the border collie variants. The dog showed an excellent clinical response to oral cobalamin supplementation with no relapse after 4 months. In conclusion, this case creates awareness that Imerslund-Gräsbeck syndrome should be considered even in mixed-breed dogs with compatible clinical signs and that two different pathogenic CUBN mutations in compound heterozygosity can lead to a typical Imerslund-Gräsbeck syndrome phenotype.     

Use of nootkatone as a senescence indicator for Rouge La Toma Cv. grapefruit (Citrus paradisi Macf.)

The objective of this research was to study the usefulness of nootkatone as a senescence indicator for Rouge La Toma cv. grapefruit (Citrus paradisi Macf.), simulating different treatments that included the normal postharvest handling of citrus fruits: temperature conditioning, cold storage, shipment periods to overseas markets such as Japan and the U.S., marketing conditions, and storage at nonchilling temperature (control treatments). The highest nootkatone levels, determined by GLC-MS analyses, were detected in fruits subjected to control treatments. No significant differences were observed in nootkatone levels between treatments either with or without temperature conditioning prior to the start of the cold storage. Levels of nootkatone increased throughout time for all assayed treatments. The linear regressions of nootkatone levels showed correlation coefficients of 0.80 and 0.83 with storage time (29 and 42 days, respectively). Therefore, nootkatone appears to be a good indicator of senescence for Rouge La Toma grapefruit.     

Evaluation in mice of Brucella ovis attenuated mutants for use as live vaccines against B. ovis infection

Brucella ovis causes ram contagious epididymitis, a disease for which a specific vaccine is lacking. Attenuated Brucella melitensis Rev 1, used as vaccine against ovine and caprine brucellosis caused by B. melitensis, is also considered the best vaccine available for the prophylaxis of B. ovis infection, but its use for this purpose has serious drawbacks. In this work, two previously characterized B. ovis attenuated mutants (Δomp25d and Δomp22) were evaluated in mice, in comparison with B. melitensis Rev 1, as vaccines against B. ovis. Similarities, but also significant differences, were found regarding the immune response induced by the three vaccines. Mice vaccinated with the B. ovis mutants developed anti-B. ovis antibodies in serum of the IgG₁, IgG_2a and IgG_2b subclasses and their levels were higher than those observed in Rev 1-vaccinated mice. After an antigen stimulus with B. ovis cells, splenocytes obtained from all vaccinated mice secreted similar levels of TNF-α and IL12(p40) and remarkably high amounts of IFN-γ, a crucial cytokine in protective immunity against other Brucella species. By contrast, IL-1α -an enhancer of T cell responses to antigen- was present at higher levels in mice vaccinated with the B. ovis mutants, while IL-10, an anti-inflammatory cytokine, was significantly more abundant in Rev 1-vaccinated mice. Additionally, the B. ovis mutants showed appropriate persistence, limited splenomegaly and protective efficacy against B. ovis similar to that observed with B. melitensis Rev 1. These characteristics encourage their evaluation in the natural host as homologous vaccines for the specific prophylaxis of B. ovis infection.     

Determination of thiamin in cooked sausages

A reliable and rapid high-performance liquid chromatography (HPLC) method has been set up for the determination of total thiamin in difficult sample matrices such as cooked sausages. Different hydrolysis conditions and enzymes were tested to release the vitamin from its phosphate ester. The best data in the enzymatic digestion were obtained by incubating the samples with 6% clara-diastase at 50 degrees C for 3 h. After oxidation of thiamin to thiochrome, the sample extracts were purified by using a C(18) Sep-Pak cartridge. Final determination was performed by reversed-phase HPLC with fluorescence detector (excitation 360 nm, emission 430 nm), on a low-cost 25 cm x 4 mm i.d. Spherisorb C(8) cartridge using a mixture of 5 mM phosphate buffer pH 7.0 and acetonitrile (70:30, v/v) as mobile phase. Precision of the method was 1.5% (within a day) and 5. 2% (between days). The detection limit was 0.015 mg/100 g. All the recoveries from the different cooked sausages were better than 90% of thiamin hydrochloride added to samples of meats. In the samples analyzed, the mean value for thiamin was between 0.039 and 0.508 mg/100 g fresh weight.     

Geomorphological significance of lichen colonization in a present snow hollow: Hoya del Cuchillar de las Navajas, Sierra de Gredos (Spain)

This paper discusses the results of a lichenometrical and geomorphological study of one of the few remaining active snow hollows in the central region of the Iberian Peninsula. The study area, located on a glacial shoulder, is called Hoya del Cuchillar de las Navajas. A protalus rampart occurs at the base of the hollow. Our studies, conducted between 1992 and 1998, were designed to determine the geomorphological characteristics of Hoya, the mobility of the deposits, and the characteristics of the snow cover. These data formed the basis for a study of the lichen colonization on the blocks and on the wall surrounding the snow hollow. All of the lichen species found were analyzed according to their abundance, distribution and the extent of their surface cover. Measurements of the diameter of the thalli of the species Rhizocarpon geographicum were also obtained. Thalli of this species were found to require a mean snow-free growing season of at least 95 days (13.5 weeks) per year. Maximum mean thallus diameters indicate that the protalus rampart was formed during the Little Ice Age and became inactive 130 years ago.     

Histochemical study of the interstitial tissue in scrotal and abdominal boar testes.

The present study describes the glycosidic content of the interstitial tissue in testes from healthy boars and from unilateral and bilateral abdominal cryptorchid boars using lectin histochemistry. The Leydig cells of healthy boars contained glycans with fucosyl, mannosyl, glucosyl, neuraminic acid and galactosyl residues, which have structural and transport functions, and participate in androgen synthesis and in cell regulation. Unilateral cryptorchidism induced high glucosyl and low galactosyl content in the Leydig cells of scrotal testes, resulting in impaired androgen production. In abdominal testes, the Leydig cells exhibited increased amounts of glucosyl and reduced amounts of galactosyl and neuraminic acid residues, resulting in defective cell regulation and lack of androgen synthesis. In healthy boars, the extracellular glycans contained fucosyl, galactosyl, glucosyl and neuraminic acid residues, which confer viscoelasticity on the interstitial tissue and participate in substrate transport, hormone binding and cell-cell interaction. Unilateral cryptorchidism did not induce anomalies in extracellular glycans in scrotal testes, but unilateral and bilateral cryptorchidism resulted in an increased content of fucosyl and galactosyl, and a decreased content of glucosyl and neuraminic acid residues in abdominal testes, leading to reduced viscoelasticity and defective substrate transport across the extracellular matrix.     

Determination of total riboflavin in cooked sausages.

A simple and rapid method for determining riboflavin content in cooked sausages by ion-pair reversed-phase liquid chromatography has been set up. Samples were subjected to acid and enzymatic hydrolysis. Sample extracts were directly chromatographed, avoiding purification and concentration treatment. Final determination was performed by high-performance liquid chromatography with fluorescence detector (excitation, 227 nm; emission, 520 nm), on a 25 cm x 4 mm i.d. Spherisorb ODS-2 cartridge using a mixture of 5 mM heptanesulfonic acid adjusted to pH 2.7 with phosphoric acid and acetonitrile (75:25, v/v) as mobile phase. Precision of the method was 1.3% (within a day) and 2.6% (between days). The detection limit was 0.015 mg/100 g. The recovery was >95%.     

Soil evolution over the Quaternary period in a semiarid climate (Segre river terraces,northeast Spain)

A macro- and micromorphologic study was done on the soils from a stepped sequence of seven dated fluvial terraces in the lower Segre river valley (Lleida, northeast Spain) under a present-day semiarid Mediterranean climate. The soils have evolved from the Holocene through the early Pleistocene, providing an excellent morphostratigraphic framework for evaluating time-dependent factors influencing soil formation in a dry and calcareous environment. Throughout the chronosequence, some properties are regularly age-related specially carbonatation in subsurface horizons. The carbonates occur mainly as micrite, and although microsparite and sparite also appear in the oldest soils, they are replaced by fine-grained calcite by dissolution–reprecipitation processes (micritization process), which is active at present. Some pedological paleofeatures as the presence of sparite and recarbonated argillans in oldest terrace can be interpreted as the reflex of climatic changes during the Quaternary. In spite of this climate variability, the soils display progressive and systematic patterns of carbonate accumulation: on the lowest terraces, the soils do not yet have secondary carbonates but in the beginning Late Pleistocene calcic horizons, with carbonate pendents, are developed; these pendents increase its thickness with age although pendent growth rates decreases from Late Pleistocene to Middle Pleistocene. In the middle of the Late Pleistocene, calcic horizons evolved to petrocalcic horizons, which increase its thickness in the Middle and specially in Early Pleistocene. The presence of calcic and petrocalcic horizons is the primary basis for soil classification. This criterion is applicable not only to the soils of the lower Segre river basin, but also to many soils throughout the semiarid Mediterranean region.