Identification of c-kit mutations-independent neoplastic cell proliferation of canine mast cells

Gain-of-function mutations in the proto-oncogene c-kit have been considered the molecular mechanism of neoplastic proliferation of mast cells. However, the importance of c-kit gene mutations is not well evaluated in canine mast cell tumors (MCTs). In the present study, we established and characterized a mast cell line, HRMC, derived from a dog with MCT. We also examined c-kit mutations in HRMC cells and assessed an inhibitory effect of a tyrosine kinase inhibitor, STI571, on HRMC cells. HRMC cells had cytoplasmic metachromatic granules, chymase and tryptase, and expressed both KIT and FcepsilonRI on the cell surface. HRMC cells contained histamine and released beta-hexosaminidase through FcepsilonRI cross-linking and calcium ionophore stimulation. Nucleotide sequence analysis demonstrated no mutations in an open reading frame of c-kit cDNA and genomic DNA of the juxtamembrane domain of c-kit in HRMC cells. STI571 did not show any inhibitory effects on the proliferation of HRMC cells. These findings clearly demonstrated the existence of c-kit mutations-independent neoplastic canine mast cell proliferation. The growth factor-independent mast cell line established in this study might be valuable to explore novel mechanisms of c-kit mutations-independent neoplastic proliferation of mast cells in dogs.     

Autoimmune disease and impaired uptake of apoptotic cells in MFG-E8-deficient mice

Apoptotic cells expose phosphatidylserine and are swiftly engulfed by macrophages. Milk fat globule epidermal growth factor (EGF) factor 8 (MFG-E8) is a protein that binds to apoptotic cells by recognizing phosphatidylserine and that enhances the engulfment of apoptotic cells by macrophages. We report that tingible body macrophages in the germinal centers of the spleen and lymph nodes strongly express MFG-E8. Many apoptotic lymphocytes were found on the MFG-E8-/- tingible body macrophages, but they were not efficiently engulfed. The MFG-E8-/- mice developed splenomegaly, with the formation of numerous germinal centers, and suffered from glomerulonephritis as a result of autoantibody production. These data demonstrate that MFG-E8 has a critical role in removing apoptotic B cells in the germinal centers and that its failure can lead to autoimmune diseases.     

Thyroid hormones concentrations and ECG picture in the dog

Disorders of the thyroid gland activity are the most commonly encountered disturbances of endocrine origin in the dog. Hypo- or hyperthyroidism may disturb the function of the cardiovascular system and cause arrhythmias. The aim of this study was to evaluate the influence of thyroid gland activity on electrocardiogram (ECG) picture in the dog by comparing ECG curves of healthy dogs, dogs with hypothyroidism and dogs with cardiac insufficiency caused by endocardiosis of the mitral valve. The study was performed on 38 dogs, patients of the Department of Internal and Parasitic Diseases with Clinic for Horses, Dogs and Cats in Wroc?aw. The animals were assigned to 3 groups: Group I--control group, 13 clinically healthy dogs; Group II--14 dogs with diagnosed cardiac insufficiency caused by endocardiosis of the mitral valve; Group III--11 dogs with hypothyroidism. Clinical examination of the animals was conducted according to the following pattern: anamnesis, general clinical examination, cardiological examination (ECG, USG of the heart) and laboratory analysis (triacylglycerydes, cholesterol, T3, T4, FT4). In this study, the significant influence of thyroid gland activity on ECG picture of the evaluated dogs was found. In the dogs with hypothyroidism a decrease in the sino-atrial node activity was observed, which led to decreased heart rate. In dogs with hypothyroidism, the innerheart conduction was reduced, which was demonstrated by prolongation of the P wave, QRS complex and the QT interval.     

Protective effect of anthocyanins from black soybean seed coats on UVB-induced apoptotic cell death in vitro and in vivo

UVB radiation proves to be one of the most relevant environmental risks because of its hazardous effects, such as premature skin aging and especially skin photocarcinogenesis. Anthocyanins, water-soluble pigments present in plants, are known to be powerful antioxidants that help protect plants from UV damage. In this study, we aimed at investigating the protective effect of anthocyanins from black soybean [Glycine max (L.) Merr] seed coats on UVB-induced apoptosis, and furthermore, we investigated the molecular mechanism responsible for regulation of apoptosis in vitro and in vivo. Pretreatment with anthocyanins reduced UVB-induced reactive oxygen species levels and inhibited UVB-induced apoptotic cell death through the prevention of caspase-3 pathway activation and reduction of proapoptotic Bax protein levels. UVB irradiation induced apoptotic cell death, which was inhibited by topical application of anthocyanins in hairless mice. It is concluded that anthocyanins from the seed coat of black soybeans may be useful compounds to modulate UVB-induced photoaging.     

Actions of quinolizidine alkaloids on Periplaneta americana nicotinic acetylcholine receptors

BACKGROUND: Botanical insecticides do not play a major role as crop protectants, but they are beneficial in some applications. The authors investigated the actions of naturally occurring alkaloids on insect nicotinic acetylcholine (ACh) receptors (nAChRs) by evaluating their abilities to inhibit specific binding of [³H]imidacloprid (IMI) to nerve‐cord membranes from Periplaneta americana L. Two alkaloids were also tested for their actions on nAChRs expressed by cockroach neurons using patch‐clamp electrophysiology. RESULTS: Four natural quinolizidine alkaloids (matrine, sophocarpine, cytisine and aloperine) exhibited more than 50% inhibition of [³H]IMI binding at 10 µM , although other compounds were found to have no or low inhibitory activity. The rank order of potency based on concentration–inhibition curves was cytisine > sophocarpine ≥ aloperine ≥ matrine. Patch‐clamp analysis indicated that sophocarpine and aloperine were not agonists of nAChRs expressed in P. americana neurons, yet, at 10 µM , aloperine, but not sophocarpine, suppressed ACh‐induced inward currents significantly. CONCLUSION: Three of the four natural alkaloids tested possess structural moieties that are necessary for interaction with P. americana nAChRs. Aloperine, which possesses a unique structure and showed a distinctive dose–response curve, was found to act as an antagonist. Appropriate modifications of these alkaloids might result in novel insecticidal nAChR ligands. Copyright © 2008 Society of Chemical Industry     

Chicken peripheral blood CD3+CD4-CD8- cells are regulated by endocrine and nerve systems

The existence of CD3(+)CD4(-)CD8(-) T cells in thymus and spleen has already been known. However, because of the presence of large amounts of thrombocytes in peripheral blood (PB), the proportion of CD3(+)CD4(-)CD8(-) T cells in PB has yet to be investigated. Therefore, the proportion of peripheral T cell-subsets was investigated in 6-week-old chickens. The percentage of CD3(+) cells, CD4(+) cells, CD8 alpha(+) cells, CD8 beta(+), and CD3(+)CD4(-)CD8(-) cells was 76%, 41%, 14%, 5%, and 15%, respectively. The proportion of CD3(+)CD4(-)CD8(-) cells in PB increased during egg-laying periods and in chickens treated with an analog of estrogen, while it decreased with age and in response to restraint stress. All of the CD3(+)CD4(-)CD8(-) cells expressed TCR1, and did not have NK activity. CD3(+)CD4(-)CD8(-) cells represent about 60% of peripheral TCR1(+) cells. These findings indicate that the proportion of CD3(+)CD4(-)CD8(-) cells is regulated by the endocrine and nerve systems.     

Isolation of myxoviruses from migratory waterfowls in San-in district,western Japan in winters of 1997-2000

Between November 1997 and February 2000, winter migratory waterfowls of several species staying in San-in district, western Japan were surveyed for influenza A virus and paramyxovirus at four stations. A total of 18 influenza A viruses was isolated from 1,404 fecal samples of whistling swans, pintails, mallards, and white-fronted geese. Five different hemagglutinins and eight neuraminidases were identified in the viruses isolated, in 11 different combinations, including H7N8 related to a subtype of a highly pathogenic chicken virus. In 2000, five lentogenic (non-pathogenic) Newcastle disease viruses were also isolated from white-fronted geese. These results suggested that possible precursor viruses for highly pathogenic avian myxoviruses are still brought into Japan by migratory waterfowls. The results also support the contention that continued surveillance of wild waterfowl population should be an integral part of control policies for these serious poultry diseases.     

Current rectification by Pauli exclusion in a weakly coupled double quantum dot system

We observe spin blockade due to Pauli exclusion in the tunneling characteristics of a coupled quantum dot system when two same-spin electrons occupy the lowest energy state in each dot. Spin blockade only occurs in one bias direction when there is asymmetry in the electron population of the two dots, leading to current rectification. We induce the collapse of the spin blockade by applying a magnetic field to open up a new spin-triplet current-carrying channel.     

A role for interaction of the RNA polymerase flap domain with the sigma subunit in promoter recognition

In bacteria, promoter recognition depends on the RNA polymerase sigma subunit, which combines with the catalytically proficient RNA polymerase core to form the holoenzyme. The major class of bacterial promoters is defined by two conserved elements (the -10 and -35 elements, which are 10 and 35 nucleotides upstream of the initiation point, respectively) that are contacted by sigma in the holoenzyme. We show that recognition of promoters of this class depends on the "flexible flap" domain of the RNA polymerase beta subunit. The flap interacts with conserved region 4 of sigma and triggers a conformational change that moves region 4 into the correct position for interaction with the -35 element. Because the flexible flap is evolutionarily conserved, this domain may facilitate promoter recognition by specificity factors in eukaryotes as well.