Characterization of cytoplasmic hyaline bodies in a hepatocellular carcinoma of a dog

This report describes the morphological and immunohistochemical features of intracytoplasmic inclusion bodies found in a 13-year-old Yorkshire dog with a hepatocellular carcinoma and referred for anorexia, lethargy and mild polydipsia. Fine-needle aspirates of the large abdominal mass revealed high number of pleomorphic neoplastic hepatocytes, containing round to polygonal, well-demarcated, hyaline bodies. Same findings were histologically confirmed on multiple biopsies. Immunohistochemically, the inclusion bodies were negative for alpha-1-antitrypsin, carcinoembryonary antigen, fibrinogen, IgG, IgM, cytokeratins 7, 8, 18, 19, 20. By transmission electron microscopy, the cytoplasmic inclusions were composed of granular homogeneous or reticulated electrondense matrix, enclosed within dilated rough endoplasmic reticulum or remnants of its membranes, consistent with proteinaceous material accumulated within neoplastic hepatocytes due to aberrant protein secretion or transport. This is the first detailed characterization of hyaline cytoplasmic inclusion bodies in canine hepatocellular carcinoma.     

Survival of the biocontrol agents Coniothyrium minitans and Bacillus subtilis MBI 600 introduced into pasteurised, sterilised and non-sterile soils

The biocontrol agents Coniothyrium minitans and Bacillus subtilis MBI 600 were added separately to three soil types that had been either sterilised, pasteurised or left non-sterile. Applied as a conidial suspension of 1×10⁶ cfu g⁻¹ soil, C. minitans showed good survival in all sterilised, pasteurised and non-sterile soils, remaining at the numerical level at which it was applied for the duration of the 30 d experiment. Applied at a lower rate of 1×10³ cfu g⁻¹ soil, C. minitans proliferated in sterilised soil to numbers slightly over 1×10⁶ cfu g⁻¹ soil, whereas no increase was seen in pasteurised or non-sterile soils from this lower application rate. However, although C. minitans was not easily recovered on plates from non-sterile soil, it did survive at the lower numerical level in pasteurised soil, and was recoverable throughout the experiment at the rate at which it was applied. B. subtilis MBI 600 survived well following introduction as a cell suspension into sterilised soil at a rate of 1×10⁶ cfu g⁻¹ soil. Spores were formed rapidly and, after 14 d, the introduced microorganism survived in this form rather than as vegetative cells. However, in non-sterile soil, the introduced microorganism did not compete well and decreased in number, with spores being formed in low numbers. Survival of B. subtilis MBI 600 in pasteurised soil was variable, but resembled the survival seen in non-sterile soil more than that seen in sterilised soil. More B. subtilis MBI 600 spores were formed in pasteurised soil than in non-sterile soil, however, and may have been important for survival in pasteurised soil. In conclusion, this work has shown that the biocontrol agent C. minitans can survive well in soil irrespective of whether the soil has been pasteurised or not and shows good promise as a soil inoculant for control of Sclerotinia sclerotiorum. Although soil pasteurisation does improve establishment of B. subtilis MBI 600 compared to non-sterile soil, survival is relatively poor when applied as cells. The best survival of B. subtilis MBI 600 occurred as spores in sterilised soil, and spore applications to pasteurised soil in an integrated control strategy may allow sufficient establishment of the biocontrol agent to target pathogens causing damping-off.     

Survival of Coniothyrium minitans associated with sclerotia of Sclerotinia sclerotiorum in soil

The development and survival of the mycoparasite Coniothyrium minitans associated with sclerotia of the plant pathogen Sclerotinia sclerotiorum was studied in pasteurised and non-sterile (untreated) soil. Using scanning electron microscopy, developing pycnidia were first seen within the sclerotial medulla at 7 days post-inoculation with the mycoparasite in pasteurised soil. However, by 14 days post-inoculation, pycnidia had developed fully in both pasteurised and non-pasteurised treatments, and conidial droplets were exuded onto the outer surface of the infected sclerotia. Thirty days post-inoculation, irrespective of soil treatment, the majority of the sclerotial medulla had been converted to pycnidia, with the sclerotial rind remaining largely intact. The pycnidia and dried intact droplets were still observed 6 months post-inoculation with C. minitans, although the conidia on the outer surface of the dried droplets had largely collapsed by this stage. Germinability studies at 10 months post-inoculation showed that approximately 13% of the conidia in dried droplets were still viable. This work shows the potential for infected sclerotia of S. sclerotiorum to provide a unique reservoir for the survival of C. minitans.     

Headspace sorptive extraction (HSSE), stir bar sorptive extraction (SBSE), and solid phase microextraction (SPME) applied to the analysis of roasted Arabica coffee and coffee brew.

Headspace sorptive extraction (HSSE) and stir bar sorptive extraction (SBSE), two recently introduced solventless enrichment techniques, have been applied to the analysis of the headspace of Arabica roasted coffee and of the headspace of the brew and of the brew itself. In both HSSE and SBSE enrichment is performed on a thick film of poly(dimethylsiloxane) (PDMS) coated onto a magnet incorporated in a glass jacket. Sampling is done by placing the PDMS stir bar in the headspace (gas phase extraction or HSSE) or by immersing it in the liquid (liquid phase extraction or SBSE). The stir bar is then thermally desorbed on-line with capillary GC-MS. The performance of HSSE and SBSE have been compared through the determination of the recoveries and relative abundances of 16 components of the coffee volatile fraction to classical static headspace (S-HS) and to headspace and in-sample solid phase microextraction (HS-SPME and IS-SPME, respectively) applying the fibers PDMS 100 microm, Carbowax/divinylbenzene 65 microm (CW/DVB), Carboxen/PDMS 75 microm(CAR/PDMS), polyacrylate 85 microm(PA), PDMS/divinylbenzene 65 microm(PDMS/DVB), and Carboxen/divinylbenzene/PDMS 50-30 microm(CAR/PDMS/DVB). In all cases, HSSE and SBSE gave higher recoveries, and this is entirely due to the high amount of PDMS applied.     

Presence of allergenic proteins in different peach (Prunus persica) cultivars and dependence of their content on fruit ripening

It has been reported that various cultivars of fruits and vegetables may present a different pattern for the contained allergens. Here, we report on the different content in allergenic proteins for different peach (Prunus persica) cultivars, sampled during two consecutive harvest seasons. Fruits from six cultivars of peaches were harvested fully ripe, and the proteins extracted from whole or chemically peeled fruits were analyzed by SDS-PAGE and immunoblotting. All the protein extracts from whole fruit contained a 9 kDa protein. This protein proved to be absent in the extracts taken from chemically peeled fruit. In four cultivars, this protein corresponds to the allergen Pru p3, a lipid transfer protein that causes the oral allergy syndrome (OAS) in sensitized people. In the following year, fruits from four of the six cultivars of peaches studied previously were harvested at different times, at one and two weeks before the commercial ripening time and when fully ripe, to ascertain whether the presence of the 9 kDa allergen might be related to the ripening process. Two cultivars out of four produced an intense allergenic band corresponding to a 9 kDa protein already two weeks before the commercial ripening date, while the others showed a progressive increment of the 9 kDa allergen during ripening.     

Erratum to: Comparative analysis of a wood-adhesive bondline

The wood–adhesive interface was analyzed using five methods with the objective of quantitatively assessing penetration of adhesive into the porous wood network. Methods included fluorescence microscopy, scanning electron microscopy, backscatter electron imaging, wavelength dispersive spectroscopy, and X-ray microtomography (XMT). Each method provided a visual inspection, and all of the analysis methods were applied to the same field of view. XMT was the primary technique of interest. Rubidium hydroxide was used in place of sodium hydroxide in the formulation of phenol–formaldehyde adhesive. Rubidium was found to increase the X-ray attenuation of the adhesive. However, rubidium migrated beyond the adhesive interphase during specimen preparation, thus reducing its effectiveness for image contrast enhancement. The wood species studied included red oak (Quercus rubra), Douglas-fir (Pseudotsuga menziesii), and hybrid poplar (Populus deltoides × Populus trichocarpa). All techniques used for this study were useful, but each presented some limitations for bondline analysis. Despite the problem with rubidium migration, XMT for this application was promising.     

Locomotor primitives in newborn babies and their development

How rudimentary movements evolve into sophisticated ones during development remains unclear. It is often assumed that the primitive patterns of neural control are suppressed during development, replaced by entirely new patterns. Here we identified the basic patterns of lumbosacral motoneuron activity from multimuscle recordings in stepping neonates, toddlers, preschoolers, and adults. Surprisingly, we found that the two basic patterns of stepping neonates are retained through development, augmented by two new patterns first revealed in toddlers. Markedly similar patterns were observed also in the rat, cat, macaque, and guineafowl, consistent with the hypothesis that, despite substantial phylogenetic distances and morphological differences, locomotion in several animal species is built starting from common primitives, perhaps related to a common ancestral neural network.     

Zorrimidazolone, a bioactive alkaloid from the non-indigenous mediterranean stolidobranch Polyandrocarpa zorritensis

Chemical analysis of the Mediterranean ascidian Polyandrocarpa zorritensis (Van Name 1931) resulted in the isolation of a series of molecules including two monoindole alkaloids, 3-indolylglyoxylic acid (3) and its methyl ester (4), 4-hydroxy-3-methoxyphenylglyoxylic acid methyl ester (1) and a new alkaloid we named zorrimidazolone (2). The structure of the novel compound 2 has been elucidated by spectroscopic analysis and bioactivity of all compounds has been investigated. Zorrimidazolone (2) showed a modest cytotoxic activity against C6 rat glioma cell line.     

Slow fertilization of stickleback eggs: the result of sexual conflict?

Background  

The fertilization success in sperm competition in externally fertilizing fish depends on number and quality of sperm. The time delay between sequential ejaculations may further influence the outcome of sperm competition. Such a time interval can load the raffle over fertilization if fertilization takes place very fast. Short fertilization times are generally assumed for externally fertilizing fish such as the three-spined stickleback (Gasterosteus aculeatus). In this pair-spawning fish, territorial males often try to steal fertilizations in nests of neighbouring males. This sneaking behaviour causes sperm competition. Sneakers will only get a share of paternity when eggs are not fertilized immediately after sperm release. Contrary to males, females may be interested in multiple paternity of their clutch of eggs. There thus may be a sexual conflict over the speed of fertilization.