Luteal and follicular count in bitches: assessment by means of magnetic resonance imaging

This study was done to determine whether preovulatory follicles or corpora lutea (physiological structures, PS) can be counted in the ovaries of bitches by means of magnetic resonance imaging (MRI). In Experiment 1, the ovaries from 15 German shepherd bitches (five in the follicular phase, one in the periovulatory period, five during the first 38 days of diestrus and four between Day 48 of diestrus and full-term gestation) were embedded in gelatin to form three phantoms with 10 ovaries each. Each phantom was exposed to MRI, using a 1mm slice thickness, a 1mm slice interval, a voxel size of 1mm cubic and a variety of pulse sequences, whereafter the ovaries were dissected and the numbers of follicles, corpora lutea and cysts counted. T2-weighted images were superior to T1-weighted images. Each of three operators counted the numbers of PS and cysts on T2-weighted images obtained in the coronal, transverse and sagital planes of each ovary, which, for the 30 ovaries, provided 270 operator by ovary by plane estimations and 90 operator by ovary estimations for each type of structure. Images of cysts were hyperintense, those of early corpora lutea and follicles similar and moderate and those of late corpora lutea hypo-intense and not clearly discernable from ovarian stroma. Estimations of PS were too low in 68%, correct in 12% and too high in 20% of estimations (n=270). Estimations of PS were correct in three operator by ovary combinations, out by 1 in 22 and out by more than 1 in 65. No operator estimated PS correctly in any bitch. In Experiment 2 MRI was done on three deeply sedated bitches in the periovulatory phase in an attempt to obtain images of the ovaries in order to count the follicles. The acquisition time of 5-7 min rendered images of poor quality from live bitches and none of their ovaries could be seen. MRI is not suitable for counting follicles or corpora lutea in the ovaries of bitches.     

Nisin inducible production of listeriolysin O in <Emphasis Type="Italic">Lactococcus lactis</Emphasis> NZ9000

Background  

Listeria monocytogenesis a well-characterized food-borne pathogen that infects pregnant women and immunocompromised individuals. Listeriolysin O (LLO) is the major virulence factor of the pathogen and is often used as a diagnostic marker for detection of L. monocytogenes. In addition, LLO represents a potent antigen driving T cell-mediated immunity during infection. In the present work, Lactococcus lactisNZ9000 was used as an expression host to hyper-produce LLO under inducible conditions using the NICE (NIsin Controlled Expression) system. We created a modified pNZ8048 vector encoding a six-His-tagged LLO downstream of the strong inducible PnisA promoter.     

Seed morphology,presence of areoles and water entry at imbibition of <i>Vicia dasycarpa</i> Ten. cv. Tolse F.C.A

The morphological characteristics of the seeds of Vicia dasycarpa Ten. cv. Tolse FCA were studied in relation to the area of imbibition water entry and were considered the presence of areoles. Seeds were analyzed using a stereo, optical and scanning electron microscope (SEM). The determination of the initial water entry area was carried out by immersing the seeds in a solution of tetrazolium (1%). This study showed that this species has seeds with a halo framing the hilum, an inconspicuous dry aril and a deltoid micropyle. The seedcoat pattern is papillose. The tracheid bar is surrounded by a ring of parenchymatous cells, and the tracheids show slight warty and non-vestured pits. It was confirmed the presence of an endospermic radicle pocket that surrounds and protect the radical tip. Two pairs of cotyledonar areoles were identified. It was established that the entry of water during imbibition starts in the area of the lens -having cracks- and moves in the sagittal plane. Both citological characteristic of tracheid bar and areoles presence show an apomorfous state between the Papiplionoids.     

A simple method for studying whole sections of rice grain

We developed a new and simple method to collect sections of a whole brown rice kernel for investigation of histological properties. A single kernel of rice was dehydrated through a graded ethanol series, transferred to xylene, and embedded in paraffin. During sectioning of the blocks using a rotary microtome, we used a special adhesive tape to collect and place the sections on slides so they remained flat. The use of the adhesive tape technique combined with autofluorescence characteristics allowed us to visualize cell walls throughout an entire longitudinal or transverse section of a whole rice kernel. We obtained scanning electron microscopy images of the sections to determine section quality.     

Staphylococcus aureus and Wegener's granulomatosis

Wegener's granulomatosis (WG) is a form of systemic vasculitis. It is characterized by granulomatous inflammation in the upper and lower airways, vasculitis and necrotizing glomerulonephritis, and is strongly associated with antineutrophil cytoplasmic antibodies against proteinase 3. Since the etiology of the disease is not clear, treatment, consisting of corticosteroids and immunosuppressives, is nonspecific and associated with severe side effects. Pinpointing the trigger(s) of the disease would highly improve treatment. Clinical evidence shows that an infectious agent, the bacterium Staphylococcus aureus, is a risk factor for disease relapse, suggesting its involvement in the pathogenesis of WG. Here we review both clinical and experimental data that either indicate or support a role for S. aureus in WG.     

Autoantibodies directed to novel components of the PM/Scl complex, the human exosome

The autoantigenic polymyositis/scleroderma (PM/Scl) complex was recently shown to be the human homologue of the yeast exosome, which is an RNA-processing complex. Our aim was to assess whether, in addition to targeting the known autoantigens PM/Scl-100 and PM/Scl-75, autoantibodies also target recently identified components of the PM/Scl complex. The prevalence of autoantibodies directed to six novel human exosome components (hRrp4p, hRrp40p, hRrp41p, hRrp42p, hRrp46p, hCsl4p) was determined in sera from patients with idiopathic inflammatory myopathy (n = 48), scleroderma (n = 11), or the PM/Scl overlap syndrome (n = 10). The sera were analyzed by enzyme-linked immunosorbent assays and western blotting using the affinity-purified recombinant proteins. Our results show that each human exosome component is recognized by autoantibodies. The hRrp4p and hRrp42p components were most frequently targeted. The presence of autoantibodies directed to the novel components of the human exosome was correlated with the presence of the anti-PM/Scl-100 autoantibody in the sera of patients with idiopathic inflammatory myopathy (IIM), as was previously found for the anti-PM/Scl-75 autoantibody. Other clear associations between autoantibody activities were not found. These results further support the conception that the autoimmune response may initially be directed to PM/Scl-100, whereas intermolecular epitope spreading may have caused the autoantibody response directed to the associated components.     

Human elongation factor 1 beta: cDNA and derived amino acid sequence.

From a cDNA library in lambda gt11 derived from poly (A+)RNA of human ovarian granulosa cells a cDNA clone lambda HGP34, containing an EcoRI insert of 829 bp, was identified. After subcloning of the insert into pUC18, the clone pHGP34 was obtained and sequenced. The derived amino acid sequence, corresponding to a protein of 225 amino acids, shows a high degree of homology to elongation factor 1 beta (EF-1 beta) of Artemia salina (57%) and known peptide sequences of Xenopus laevis EF-1 beta (86%). We therefore assume that the protein coded for by pHGP34 represents human EF-1 beta. Northern analysis reveals an EF-1 beta specific mRNA of 900 bp. Southern analysis indicates that EF-1 beta in the human genome, like EF-1 alpha, appears to be specified by more than one gene. A high degree of sequence homology for EF-1 beta specific sequences is observed for bovine, rat and mouse species.     

Seminalplasmin, the major basic protein of bull seminal plasma, is a secretory protein of the seminal vesicles.

From the experimental results of three independent methods: (1) indirect immunofluorescence employing monospecific anti-seminalplasmin-IgGs, (2) cell-free translation of poly(A)+ RNA from seminal vesicle and testicular tissue, as well as (3) Northern analysis of poly(A)+ RNA of the latter tissues with a synthetic seminalplasmin-specific antisense DNA probe, it is concluded that the biosynthesis of seminalplasmin occurs in seminal vesicles but not in testis.     

Polymorphism and divergence at the prune locus in Drosophila melanogaster and D. simulans

The prune locus of Drosophila melanogaster lies at the tip of the X chromosome, in a region of reduced recombination in which nearby loci show reduced variation relative to evolutionary divergence from D. simulans. DNA sequencing of prune alleles from D. melanogaster and D. simulans reveals extremely low variation in D. melanogaster but greater variation in D. simulans. Divergence between the two species is not reduced. This pattern may be explained by either positive selection leading to hitchhiking of neutral variation or background selection against deleterious mutations. The pattern of silent versus replacement polymorphism and divergence at prune is consistent with either a model of weakly deleterious selection against amino acid substitutions or balancing selection.