Molecular characteristics of cutaneous papillomavirus from the canine pigmented epidermal nevus

To investigate the relation between the canine pigmented epidermal nevus (PEN) and cutaneous papillomavirus, we cloned and sequenced the L1 gene of papillomavirus from the canine pigmented epidermal nevus (PEN). Amplification of DNA sample with the L1 consensus primers yielded an expected fragment of approximately 450-bp. The nucleotide sequences of the fragment showed about 64% of sequence similarity to the L1 region of human papillomavirus isolate CP6108 and less than 57% sequence similarity to those of canine oral papillomavirus (COPV). In situ hybridization determined the presence of papillomavirus DNA mainly in the upper stratum granulosum of skin in this case. The results indicated that the canine cutaneous papillomavirus from the PEN lesion was genetically close to human papillomavirus rather than COPV.     

Phylogenetic Analysis of Anaplasma marginale Strains from Paraná State,Brazil, Using the msp1α and msp4 genes

Anaplasma marginale is an obligate intraerythrocytic rickettsial pathogen (order, Rickettsiales: family, Anaplasmataceae) that causes bovine anaplasmosis. This disease is widely distributed in tropical and sub‐tropical regions of the world and causes important economic losses to cattle production. Major surface protein (MSP)1a (msp1α gene) is one of the six MSPs identified on A. marginale from cattle, whose sequence and size vary according to the number of tandem 28‐ to 29‐amino acid repeats. This study characterized the msp1α and msp4 genes obtained from three distinct Brazilian herds from the State of Paraná. Three strains of the msp1α and one strain of the msp4 gene were sequenced. The strains evaluated revealed PCR products of different size, representing three, five and six internal repeats. Sequence analyses confirmed the number of tandem sequence copies and revealed a high degree of sequence identity with strains from other Brazilian States, as well as strains from the USA, Europe and Israel. The msp1α DNA and amino acid sequences from A. marginale and DNA sequences of msp4 strains did not reveal distinct phylogeographical segregation. However, the amino acid sequences of msp4 demonstrated definite phylogeographical relationship. These results suggest that the amino acid sequences of msp4 should be used for phylogenetic identification of A. marginale strains and may be an important tool for the epidemiology and control of anaplasmosis. Additionally, the close similarity of the Paraná strains of A. marginale with strains from USA, Europe and Asia may reflect the introduction of these genes during the development of the Brazilian bovine herd.     

Immune response of BALB/c mouse immunized with recombinant MSPs proteins of Anaplasma marginale binding to immunostimulant complex (ISCOM)

Anaplasmosis, caused by Anaplasma marginale, results in significant economic losses of cattle in tropical and subtropical regions worldwide. Six major surface proteins (MSPs) were well characterized and designated as MSP1, MSP2, MSP3, MSP4, and MSP5. The objective of this study was to evaluate the humoral immune response of BALB/c mice against the recombinant MSPs, incorporated into immunostimulating complex (ISCOM). The recombinant proteins purified by Ni-NTA columns were incorporated into ISCOM and ISCOMATRIX by the lipid film hydration method. BALB/c mice immunized with ISCOM/rMSPs and ISCOMATRIX/rMSPs vaccines produced whole IgG, IgG1, and IgG2a, in contrast to the negative groups (PBS and ISCOMATRIX adjuvant). All groups that received antigen responded specifically against the rMSPs by Western blotting, showing the rMSP1a (60-105kDa), rMSP1b (100kDa), rMSP4 (47kDa), and rMSP5 (29kDa). Additional studies will have to be performed in cattle to evaluate the humoral and cellular mechanisms of this subunit vaccine and their possible use as protective vaccines against homologous and heterologous strains of A. marginale.     

Evidence for Alfvén waves in solar x-ray jets

Coronal magnetic fields are dynamic, and field lines may misalign, reassemble, and release energy by means of magnetic reconnection. Giant releases may generate solar flares and coronal mass ejections and, on a smaller scale, produce x-ray jets. Hinode observations of polar coronal holes reveal that x-ray jets have two distinct velocities: one near the Alfvén speed ( approximately 800 kilometers per second) and another near the sound speed (200 kilometers per second). Many more jets were seen than have been reported previously; we detected an average of 10 events per hour up to these speeds, whereas previous observations documented only a handful per day with lower average speeds of 200 kilometers per second. The x-ray jets are about 2 x 10(3) to 2 x 10(4) kilometers wide and 1 x 10(5) kilometers long and last from 100 to 2500 seconds. The large number of events, coupled with the high velocities of the apparent outflows, indicates that the jets may contribute to the high-speed solar wind.     

Slipping magnetic reconnection in coronal loops

Magnetic reconnection of solar coronal loops is the main process that causes solar flares and possibly coronal heating. In the standard model, magnetic field lines break and reconnect instantaneously at places where the field mapping is discontinuous. However, another mode may operate where the magnetic field mapping is continuous but shows steep gradients: The field lines may slip across each other. Soft x-ray observations of fast bidirectional motions of coronal loops, observed by the Hinode spacecraft, support the existence of this slipping magnetic reconnection regime in the Sun's corona. This basic process should be considered when interpreting reconnection, both on the Sun and in laboratory-based plasma experiments.     

Ascorbate regeneration by the reduced form of 2-amino-3-carboxy-1, 4-naphthoquinone, a strong growth stimulator for bifidobacteria

Nonenzymatic reduction of dehydroascorbate into ascorbate by the reduced form (quinol form) of 2-amino-3-carboxy-1,4-naphthoquinone, a strong growth stimulator for bifidobacteria, has been found. The bimolecular reaction rate constant was evaluated as 9 M(-)(1) s(-)(1) at pH 7.0. This reaction has been successfully coupled with enzymatic regeneration of the naphthoquinol by NAD(P)H in cell-free extracts of Bifidobacterium longum 6001. The overall reaction is a regeneration of NAD(P)(+) by dehydroascorbate [or a regeneration of ascorbate by NAD(P)H], in which the naphthoquinone/quinol redox couple functions as an electron transfer mediator. Kinetic study of the reduction of dehydroascorbate with related quinol compounds suggested the significance of the amino substituent of the naphthoquinol. A mechanism of the electron transfer from the quinol to dehydroascorbate is proposed, where the first step of the reaction is a nucleophilic addition of the C(2)-amino substituent of the naphthoquinol to the C(2)-position of dehydroascorbate to form a Schiff base intermediate.     

Subepithelial elastic fibers in the bovine gastric mucosa

The arrangement of subepithelial elastic fibers in the bovine gastric mucosa was studied by light and electron microscopy, with special reference to the elastic architecture of the cornified papillae. A common feature of their arrangement was that the fibers, derived from the elastic plexus in the deep layer of the lamina propria, extend perpendicularly toward the epithelium giving off the slender branches that end at the epithelial basement membrane. The connecting branches were observed electron microscopically in the form of oxytalan or slender elaunin fibers. The high frequency of fibers was found in the mucosa of the upper part of the ruminal atrium, the ruminal pillars, the reticular cristae and two kinds of cornified unguiculiform and omasal papillae. Each papilla consisted of the core of three-dimensional elastic network and numerous connecting-twigs arising from the superficial points of the network. The mucosae of the ruminal sacs, the floor of the reticular cells and the abomasum except the pyloric part were relatively poor in elastic fibers. These results suggest that the variation in fiber-arrangement may be relevant to the condition of mechanical stress inflicting on the gastric innersurface.     

Serum creatine kinase activities in dogs with dirofilariasis.

Total serum creatine kinase (CK) and its isozyme activities were determined in dogs with dirofilariasis. Before heartworm removal, total CK and isozyme activities in dogs of the mild group were not different from those in dogs of the heartworm-free group. BB activity was higher in dogs of the hemoptysis group. Dogs of the ascites group displayed a mild increase in MM activity. In dogs of the caval syndrome (CS) group, total CK and MM activities were highest among the heartworm-free and heartworm-infected dogs, and MM isozyme accounted for most (75%) of total CK activity. MB and BB activities were also higher. However, there were no significant differences in CK activities between the surviving and non-surviving cases. In dogs with pulmonary heartworm disease (mild and ascites groups), MM activity correlated significantly with the number of heartworms (r = 0.45), hematocrit value (Ht, r = -0.40), serum alanine aminotransferase (ALT, r = 0.42) and lactate dehydrogenase (LDH, r = 0.46) activities, mean pulmonary arterial pressure (r = 0.64) and total pulmonary resistance (r = 0.50). In dogs with CS, MM activity did not correlate with any parameter, but BB activity correlated with the number of heartworms at the right atrium (r = 0.61), Ht (r = -0.53), ALT (r = 0.80), LDH (r = 0.73) and serum urea nitrogen (r = 0.47). At 1 week after heartworm removal, BB and MM activity decreased in dogs of the hemoptysis and ascites groups, respectively. In dogs of the CS group, total CK and MM isozyme activities decreased markedly (P less than 0.01) regardless of their prognosis.     

Aberrant development of mammary glands, but precocious expression of beta-casein in transgenic females ubiquitously expressing whey acidic protein transgene

It has been suggested that whey acidic protein (WAP) may function as a protease inhibitor. However, the actual function of WAP remains obscure. We investigated the histological development of the mammary glands of transgenic mice ubiquitously expressing WAP and CAG/WAP transgene. Ubiquitous expression of WAP induced aberrant development of the lobular alveoli of the mammary glands: mammary alveoli that were either aberrantly large or small in size increased in number in the developing mammary glands of these transgenic females during pregnancy and lactation. The expression of beta-casein was precociously induced in the mammary glands of the transgenic females during early pregnancy and accompanying this was a histological observation that abnormally developed lobular alveoli filled with milk proteins appeared in the mammary glands of transgenic females during early pregnancy. However, during lactation, the development of mammary glands was impaired in transgenic females. To investigate the possible paracrine action of WAP associated with mammary gland aberration, we transplanted the mammary tissue of CAG/EGFP transgenic females into the fat pad of virgin CAG/WAP transgenic females and initiated pregnancy by mating. The development of mammary tissue transplanted to the recipient was histologically examined on day 3 of lactation. The results revealed that the development of grafted mammary tissues was impaired in a manner similar to that of the mammary glands of CAG/WAP transgenic females, indicating that the inhibitory effect of WAP acts via a paracrine mechanism. In vitro experiments using HC11 cells with forced expression of exogenous WAP demonstrated the inhibitory function of WAP on proliferation of mammary epithelial cells.