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Background: We recently reported that multilineage-differentiating stress enduring (Muse) cells intravenously administered after acute myocardial infarction (AMI), selectively engrafted to the infarct area, spontaneously differentiated into cardiomyocytes and vessels, reduced the infarct size, improved the left ventricular (LV) function and remodeling in rabbits. We aimed to clarify the efficiency of Muse cells in a larger animal AMI model of mini-pigs using a semi-clinical grade human Muse cell product. Method and result: Mini-pigs underwent 30 min of coronary artery occlusion followed by 2 weeks of reperfusion. Semi-clinical grade human Muse cell product (1x107, Muse group, n = 5) or saline (Vehicle group, n = 7) were intravenously administered at 24 h after reperfusion. The infarct size, LV function and remodeling were evaluated by echocardiography. Arrhythmias were evaluated by an implantable loop recorder. The infarct size was significantly smaller in the Muse group (10.5±3.3%) than in the Vehicle group (21.0±2.0%). Both the LV ejection fraction and fractional shortening were significantly greater in the Muse group than in the Vehicle group. The LV end-systolic and end-diastolic dimensions were significantly smaller in the Muse group than in the Vehicle group. Human Muse cells homed into the infarct border area and expressed cardiac troponin I and vascular endothelial CD31. No arrhythmias and no blood test abnormality were observed. Conclusion: Muse cell product might be promising for AMI therapy based on the efficiency and safety in a mini-pig AMI. [ABSTRACT FROM AUTHOR]

To understand the anatomical characteristics of microminipigs, one of the smallest miniature pigs, as a large animal model, we measured the body and organ sizes of four-, five-, six-, and seven-month-old microminipigs (n = 4, females) using computed tomography. In addition, the results were compared with those of young mature beagles (10 months old, two males and three females), which have been widely used as a large animal model. The microminipigs at 4–6 months of age were much smaller than the beagles. However, when the microminipigs reached seven months of age, their overall size was similar to that of the beagles. The thoracic cavity volume of the seven-month-old microminipigs was less than half that of the beagles, and the cavity was largely filled by the heart. The liver size of the seven-month-old microminipigs was approximately half of that of the beagles. Moreover, the spleen of the seven-month-old microminipigs was different in morphology, but not different in size from that of the beagles. In addition, although their volumes were the same, the kidneys of the seven-month-old microminipigs, unlike those of the beagles, were flattened in shape. Collectively, the major abdominal organs of the seven-month-old microminipigs were either the same size or smaller than those of the beagles, but the abdominal cavity volume of the seven-month-old microminipigs was larger than that of the beagles. Thus, the abdominal cavity of microminipigs is assumed to be filled with the gastrointestinal tract. The anatomical characteristics of the young mature microminipigs revealed in our study suggest that microminipigs could have great potential as a large animal model for biomedical research. [ABSTRACT FROM AUTHOR]

A new fish reef block ensuring accurate placement by a free fall was developed to enable a more economical construction of nursery reefs for snow crabs. The block is based on a cube-shaped frame structure with extra shelf areas attached to the upper and lower frames to act as stabilizers. The falling behavior of the block in water was investigated by hydraulic model experiments and three-dimensional (3D) numerical computations. A stochastic model based on the experimental data was used to confirm a narrow distribution of the placement position as a result of the swing motion of the block during its fall. The numerical computation results indicate that the upper stabilizer produces a restoring moment for a tilted block and the lower stabilizer reduces instability in the wake of the falling block. In addition, a quick estimation method for the placement position was developed to determine the release point of the block during actual construction in the presence of ambient currents. [ABSTRACT FROM AUTHOR]

The article discusses research on suitable reference genes for quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) in total ribonucleic acid (RNA) extracted from canine whole blood using the PAXgene™ system. It says appropriate reference genes are required for determining accurately relative messenger RNA (mRNA) transcription levels, in gene expression analysis with quantitative real-time RT-PCR. It notes that a comparison of transcription levels of twelve candidate reference genes was carried out to determine the optimal reference gene in canine whole blood. It cites the use of statistical programs GeNorm, Normfinder and Bestkeeper to evaluate the reference gene's stability.

Recombinant canine interferon-γ (rCaIFN-γ) produced by a baculovirus expression system has therapeutic efficacy against atopic dermatitis in dogs. Although the mechanism of action of rCaIFN-γ is not completely understood, rCaIFN-γ is thought to downregulate the activity of interleukin-4- and interleukin-5-producing T helper 2 cells. However, rCaIFN-γ may also act directly on canine keratinocytes by inhibiting the release of inflammatory mediators. In this study, we investigated the effects of rCaIFN-γ on cytokine and chemokine mRNA transcription in a canine keratinocyte cell line, CPEK. It was found that granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA transcription was significantly inhibited after treatment with rCaIFN-γ ( P < 0.001), whereas transforming growth factor-β and CC chemokine ligand 17 mRNA levels were unchanged. This study suggests that rCaIFN-γ may suppress GM-CSF production from canine keratinocytes, although further studies are required to confirm this. [ABSTRACT FROM AUTHOR]

The article presents the results of a study on the capability of Staphylococcus pseudintermedius and Staphylococcus schleiferi taken from dogs with pyoderma in Japan to resist antimicrobial drugs and methicillin. According to the authors, isolates of coagulase-positive staphylococci (CoPS) have a high level of resistance to the antimicrobials usually used for treatment against staphylococcal infections. They add that the common cases of canine pyoderma were a high level of methicillin-resistant CoPS (MRCoPS). They also indicate remarkable differences in antimicrobial susceptibilities between staphylococcal species and between methicillin-susceptible CoPS and MRCoPS strains.

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Staphylococcal exfoliative toxins are involved in some cutaneous infections in mammals by targeting desmoglein 1 (Dsg1), a desmosomal cell-cell adhesion molecule. Recently, an exfoliative toxin gene ( exi) was identified in Staphylococcus pseudintermedius isolated from canine pyoderma. The aim of this study was to identify novel exfoliative toxin genes in S. pseudintermedius. Here, we describe a novel orf in the genome of S. pseudintermedius isolated from canine impetigo, whose deduced amino acid sequence was homologous to that of the SHETB exfoliative toxin from Staphylococcus hyicus (70.4%). The ORF recombinant protein caused skin exfoliation and abolished cell surface staining of Dsg1 in canine skin. Moreover, the ORF protein degraded the recombinant extracellular domains of canine Dsg1, but not Dsg3, in vitro. PCR analysis revealed that the orf was present in 23.2% (23/99) of S. pseudintermedius isolates from dogs with superficial pyoderma exhibiting various clinical phenotypes, while the occurrence in S. pseudintermedius isolates from healthy dogs was 6.1% (3/49). In summary, this newly found orf in S. pseudintermedius encodes a novel exfoliative toxin, which targets a cell-cell adhesion molecule in canine epidermis and might be involved in a broad spectrum of canine pyoderma. [ABSTRACT FROM AUTHOR]

Abstract: Keratinocytes produce inflammatory mediators that are involved in the pathogenesis of skin disorders such as atopic dermatitis (AD). In particular, the CC chemokines, thymus and activation regulated chemokine (TARC)/CCL17 and mucosae-associated epithelial chemokine (MEC)/CCL28 are considered to play an important role in the lesional infiltration of lymphocytes in canine AD. However, there have been no reports on the regulatory mechanisms of CCL17 and CCL28 transcription in canine keratinocytes. In this study, we investigated whether CCL17 and CCL28 transcription in cultured keratinocytes is induced by TNF-α, IL-1β, or IFN-γ. It was found that CCL17 mRNA transcription is augmented by TNF-α only, whereas the CCL28 mRNA level could be increased by TNF-α, IL-1β, or IFN-γ. The present study suggests that pro-inflammatory cytokines are important inducing factors for the production of CCL17 and CCL28 in the lesional skin of dogs with AD. [Copyright &y& Elsevier]

The article presents a study which reports on the molecular cloning of canine protease-activated receptor-2 (PAR-2) gene, and examines its transcription levels in various dog tissues and atopic skin lesions. It states that the study aim to provide better understanding of the role of PAR-2 in canine allergic diseases. It also states that quantitative real-time polymerase chain reaction (RT-PCR) was used in performing the transcription analysis of PAR-2 mRNA in normal dog tissues. Results show that PAR-2 mRNA was detected in the small intestine, colon, lung, pancreas, thyroid gland, lymph node, kidney, and skin, proving that PAR-2 can be associated to allergic inflammation and pruritus in canine atopic dermatitis (AD).

Abstract: House dust mite (HDM) allergens are the most common allergens involved in the induction of IgE-mediated hypersensitivity. Recently, epicutaneous sensitization with HDM allergens has been emphasized in the development of atopic dermatitis (AD); however, direct stimulation of canine keratinocytes by mite allergens has not been well investigated. In the present study, we investigated the effects of Der f 1, a major allergen of Dermatophagoides farinae, on cytokine and chemokine gene expression in a canine keratinocyte cell line, CPEK. CPEK constitutively expressed mRNA for TNF-α, IL-12p35, IL-18, GM-CSF, TGF-β, IL-8/CXCL8, TARC/CCL17, CTACK/CCL27 and MEC/CCL28. Of all the cytokines and chemokines investigated in CPEK, transcription levels of GM-CSF, IL-8/CXCL8 and TNF-α mRNA were significantly enhanced by stimulation with Der f 1. The present results suggest that Der f 1 can directly augment inflammatory cytokine and chemokine production from keratinocytes, and may initiate allergic inflammation independently of Type-I hypersensitivity. [Copyright &y& Elsevier]

Lipase-catalyzed acylation of ethyl (1-hydroxyalkyl)phenylphosphinates afforded a single diastereomer in high enantiomeric excess. The substituent effect of the alkyl group toward the acylation using Candida antractica (CAL) was larger than that of an immobilized lipase from Pseudomonas fluorescens lipase (lipase AK). [Copyright &y& Elsevier]