Validation of tissue microarrays for immunohistochemical analyses of canine lymphomas.

In most validation studies of tissue microarrays (TMAs), a fixed number of cores with a given diameter are analyzed to determine the degree of accuracy by which the TMA represents the whole section. The statistical model described in the present study predicts this property for various combinations of 2 core sizes (0.6 mm and 1.2 mm) and different core numbers. The model was based on artificial TMA core biopsies generated from Ki-67 and active caspase-3 immunostains of 40 canine lymphoma samples. Positivity was scored on a continuous scale, and a large number of cells were analyzed with the help of semiautomated cell counting. Despite considerable differences in range and distribution of Ki-67 and active caspase-3 positivity values, the model predictions showed a high degree of agreement for both markers. Comparison of 0.6 mm and 1.2 mm cores indicated that the use of small cores necessitates inclusion of a larger number of samples but requires counting a markedly smaller number of cells. Suitability of TMAs to determine the immunophenotype of the whole section was assessed using 2 different combinations of core sizes and numbers. Both displayed a high degree of concordance with the whole section (kappa(0.6) = 0.79; kappa(1.2) = 0.91). The present study provides a basis for the use of TMAs in future high-throughput immunohistochemical investigations of selected markers in canine lymphomas. The statistical model presented can be used to determine an optimal TMA design depending on a desired accuracy.     

Unicentric extranodal lymphoma of the upper eyelid conjunctiva in a dog

A case of conjunctival lymphoma in a 4-year-old Siberian husky is reported. A large red mass protruding from the conjunctiva of the upper eyelid of the right eye was present. Irritation, blepharospasm and epiphora were revealed on initial ophthalmic examination. After anti-inflammatory treatment, surgery was performed. Histologically, the mass consisted of large polygonal cells with a high nucleus to cytoplasm ratio, moderate amounts of slightly eosinophilic cytoplasm, pleomorphic nuclei with vesicular chromatin and prominent multiple nucleoli. Mitotic figures were frequent. Approximately 70% of the neoplastic cells were CD3 positive and CD79alpha negative. On the basis of histologic and immunohistochemical findings, a diagnosis of intermediate-grade, diffuse large-cell lymphoma with T-cell immunophenotype was made. The surgical area healed uneventfully and, although chemotherapy was not received, after 12 months the dog exhibited no recurrence. To the authors' knowledge, this is the first report of unicentric extranodal conjunctival lymphoma in a dog.     

Long-term study of the reproductive timing of the Neotropical catfish Iheringichthys labrosus (Lütken, 1874): Influence of temperature and river discharge

Unravelling the effect of climate variability on species biology has been one of the main goals of ecological studies. Environmental factors such as river discharge and temperature have being proposed as triggers of reproductive cycle in fish. In temperate climates, fish reproduction is affected mainly by temperature, while is influenced by flood pulses in large tropical rivers. We evaluated the influence of temperature and river discharge as triggers of Iheringichthys labrosus’ reproduction. We studied the following reproductive variables: gonadosomatic index (GSI), hepatosomatic index (HSI) and condition factor (K). Females of I. labrosus were examined in a time series of ten years along three sites located in Lower Uruguay River at the beginning (last week of November-first week of December, spring) and by the end of the reproductive period (middle-end of April, autumn). Generalised linear models detected a positive effect of mean winter temperature and site on the spring mature female GSI. Site was significant in addition to the interaction between temperature and site. Moreover, the effect of winter temperature was stronger than water discharge in triggering reproductive timing of I. labrosus, contrary to the model proposed for other species in tropical systems. Additionally, a literature review suggested a latitudinal gradient on reproduction, occurring earlier and prolonged with decreasing latitude (and increasing temperature). In this study, we stated the effect of temperature on the life history in a Neotropical fish using a multifaceted approach, particularly important in the current context of global climate warming.     

Effect of phase-feeding crude protein on performance and carcass characteristics of crossbred beef bulls: an application to reduce nitrogen compounds in beef cattle diets

Beef cattle (24) with an average initial body weight of 417?±?54 kg were assigned in a 2?×?2 factorial design with six repetitions. The factors studied were two levels of CP: 11 (low CP) or 13 % (high CP) in the initial feedlot phase (days 1–36) and in the final phase (days 37–72). After 36 days, half of the bulls fed with each level of CP were selected to reverse the CP level. No interaction was observed (P?>?0.05) between the level of protein in the initial and final phases on the intake of the nutrients evaluated. There was no effect (P?>?0.05) of the protein levels in the initial and final phases on intake of dry matter, organic matter, CP, non-fiber carbohydrates, and total digestible nutrients. No differences were observed among treatments (P?>?0.05) for average daily gain (X = 1.99 kg/day) and carcass traits. We conclude that it is possible to feed Holstein-Zebu crossed bulls with an average daily gain of approximately 2 kg using a fixed level of 11 % CP during the entire feedlot period, and this diet is economically viable and environmentally sound.     

Influence of cultivar on quality parameters and chemical composition of strawberry fruits grown in Brazil

Six strawberry cultivars grown on the same commercial plantation in Brazil were evaluated for their chemical composition and quality attributes at the ripe stage. The profiles of the main soluble sugars, ascorbic acid, and anthocyanins were also obtained during the developmental stages. Results showed significant differences among cultivars in all of the investigated parameters. Cv. Campineiro showed an average value for texture of 0.63 N, half the value found for cv. Oso Grande. Anthocyanin content ranged from 13 (cv. Campineiro) to 55 (cv. Mazi) mg/100 g. Total ascorbic acid found for cv. Campineiro (85 mg/100 g) was twice the amount found in cv. Dover (40 mg/100 g). Fructose was the predominant soluble sugar in almost all cultivars. The proportion among the main soluble sugars (fructose, sucrose, and glucose) was similar for Oso Grande and Toyonoka cultivars. The flavonol content (quercetin plus kaempferol derivatives) ranged from 2.7 to 7.1 mg/100 g, with a mean value of 6.1 mg/100 g, whereas free ellagic acid ranged from 0.9 to 1.9 and total phenolics varied from 159 to 289 (mean 221) mg/100 g.     

Mercury pollution in the soil and mosses around a geothermal plant

Samples of soils and mosses were collected near a geothermal power plant, which is located in one of the most active geothermal fields of the world (Southern Tuscany). High concentrations of Hg (up to 1.8 μg g^?1 d.w.) occur in mosses as far as 0.6 km from the geothermal plant, and the Hg uptake of mosses is unrelated to the species except for Bryum torquescens. The Hg in the soil was lower than in mosses, and the greatest concentrations in soil were near the Travale-22 well, which feeds the most powerful geothermal plant (30 MW) in the area. In addition, the concentrations of Hg detected in soil cores and in old bottom leaves of several specimens of Hypnum cupressiforme show that this element has been emitted into the air also in the past probably since the discovery of T-22 well in 1972.     

Molecular and physiological characterisation of psychrotrophic hydrocarbon-degrading bacteria isolated from Terra Nova Bay (Antarctica)

A set of 21 Antarctic marine bacteria isolated from the Ross Sea and able to utilise diesel fuel as the sole carbon and energy source was characterised. Isolates were analysed by amplified 16S rDNA restriction analysis using the enzyme AluI, resulting in two different groups corresponding to different bacterial species. These species were assigned to the genera Rhodococcus and Alcaligenes, on the basis of 16S rDNA sequencing. This low degree of inter-specific biodiversity was parallel to a low intra-specific biodiversity, as shown by Random Amplified Polymorphic DNA analysis. Then, a 550-bp DNA fragment coding for the inner region of alkane mono-oxygenase was PCR-amplified from the genome of each strain. The phylogenetic analysis of the sequence of the putative AlkB protein coded for by the amplified DNA fragment revealed that these alkB genes were very likely inherited by horizontal gene transfer. Lastly, the analysis of the biodegradation ability of four strains revealed two different strategies of hydrocarbon uptake, mediated either by bio-surfactants and peculiar of Rhodococcus isolates, or by membrane modifications and shown by Alcaligenes isolates. In order to understand the interrelationships between hydrocarbon-degrading isolates, the dynamics of two strains, belonging to Rhodococcus and Alcaligenes, grown together in a co-culture was also followed over a seventeen days period.     

Systemic acquired resistance in crop protection: from nature to a chemical approach

Plant natural resistance to potential parasites is regulated by two fundamental mechanisms: the "nonhost" and the "gene-for-gene" resistance, respectively. The latter is relevant when a cultivar resistant (R) gene product recognizes an avirulence gene product in the attacking pathogen and triggers an array of biochemical reactions that halt the pathogen around the site of attempted invasion. To cope with virulent pathogens, plants may benefit by some temporary immunity after a challenge triggering such an array of defense reactions, following a localized necrotizing infection as a possible consequence of a hypersensitive response (HR). This process, mediated by accumulation of endogenous salicylic acid (SA), is called systemic acquired resistance (SAR) and provides resistance, to a certain extent even against unrelated pathogens, such as viruses, bacteria, and fungi, for a relatively long-lasting period. SAR may be more potently activated in plants pretreated with chemical inducers, most of which appear to act as functional analogues of SA. This review summarizes the complex aspects of SAR as a way to prevent crop diseases by activating the plants' own natural defenses. The following outline is taken: (1) introduction through the historical insight of the phenomenon; (2) oxidative burst, which produces high levels of oxygen reactive species in a way similar to the inflammation state in animals and precedes the HR to the pathogen attack; (3) SAR as a coordinate action of several gene products leading to the expression of defenses well beyond the time and space limits of the HR; (4) jasmonic acid (JA) and ethylene as other endogenous factors mediating a different pathway of induced resistance; (5) pathogenesis related proteins (PR proteins) de novo synthesized as specific markers of SAR; (6) exogenous inducers of SAR, which include both synthetic chemicals and natural products; (7) the pathway of signal transduction between sensitization by inducers and PR expression, as inferred by mutageneses, a process that is still, to a large extent, not completely elucidated; (8) prospects and costs; (9) final remarks on the state-of-the-art of the topic reflecting the chemical view of the author, based on the more authoritative ones expressed by the authors of the reviewed papers.     

The importance of wild potato species resistant to the potato cyst nematode,Globodera pallida,pathotypes P4A and P5A,in potato breeding. I. Resistance studies

Summary Seven wild diploid potato species, Series Tuberosa, representing 1023 clones were screened for resistance to the potato cyst nematode, Globodera pallida. Over 25% of the clones were resistant to pathotype P₄A and almost 30% were resistant to pathotype P₅A. The resistance in hybrid progenies of these and other resistant species with cultivated potatoes was evaluated, and over 2200 seedlings were screened. High frequencies of resistance (>50%) to P₄A were found in progenies with Solanum leptophyes, S. vernei, S. gourlayi and S. capsicibaccatum, whereas resistance to P₅A was found in these species as well as S. sparsipilum. The importance of nematode resistant wild species for potato breeding is discussed.     

Genetic characterization of 218 elite CIMMYT maize inbred lines using RFLP markers

Characterization of genetic diversity among maize inbred lines can facilitate organization of germplasm and improve efficiency of breeding programs. A set of 218 phenotypically diverse inbred maize lines developed at CIMMYT for hybrid production was characterized using 32 RFLP markers to: (1) analyze the genetic diversity present; (2) define potential heterotic groups based on clusters formed with marker data; and (3) identify the most representative testers for each potential heterotic group. Lines were clustered using five different genetic distance measurements to find consensus non-hierarchical clusters. Dendrograms were produced to study hierarchical classification within smaller groups of lines. A very high average allelic diversity was seen in this germplasm. Lines did not cluster based on phenotype, environmental adaptation, grain color or type, maturity, or heterotic response (as determined based on hybrid performance with testers), but lines related by pedigree usually did cluster together. Previously defined testers from opposite heterotic groups were not genetically differentiated, and did not represent well their heterotic group. Discrete clusters were difficult to find; thus, potential heterotic groups will be difficult to suggest using RFLP markers alone. However, suggestions on how to use molecular markers and cross performance information to refine heterotic groups and select representative testers are presented.