Protein expression of KIT, BRAF, GNA11, GNAQ and RASSF1 in feline diffuse iris melanomas.

Feline iris melanoma, the most common feline intraocular tumour, has a reported metastatic rate of 19-63%. However, there is a lack of knowledge about its molecular biology. Previous studies have reported that feline iris melanomas do not harbour mutations comparable to common mutations found in their human counterpart. Nevertheless, there are differences in the gene expression patterns. The aim of this study was to investigate the protein expression of B-RAF oncogene serine/threonine kinase (BRAF), G protein subunit alpha q (GNAQ) and 11 (GNA11), KIT proto-oncogene receptor tyrosine kinase (KIT), and Ras association family member 1 (RASSF1) in feline iris melanomas. Fifty-seven formalin-fixed paraffin embedded (FFPE) iris melanomas and 25 FFPE eyes without ocular abnormalities were stained with antibodies against the respective proteins using immunofluorescence. Averaged pixel intensities/µm2 and percentage of stained area from total tissue area were measured and the results were compared. Compared to the control group, iris melanomas showed overexpression of BRAF, GNAQ, GNA11 and KIT. The higher expression of BRAF, GNAQ, GNA11 and KIT in feline iris melanomas suggest that these proteins may play a key role in the development of feline iris melanomas and KIT may present a possible target for future therapies in cats with feline iris melanomas.

Cobalt and manganese carboxylates for metal oxide thin film deposition by applying the atmospheric pressure combustion chemical vapour deposition process.

Coordination complexes [M(O2CCH2OC2H4OMe)2] (M = Co, 4; M = Mn, 5) are accessible by the anion exchange reaction between the corresponding metal acetates [M(OAc)2(H2O)4] (M = Co, 1; M = Mn, 2) and the carboxylic acid HO2CCH2OC2H4OMe (3). IR spectroscopy confirms the chelating or µ-bridging binding mode of the carboxylato ligands to M(ii). The molecular structure of 5 in the solid state confirms a distorted octahedral arrangement at Mn(ii), setup by the two carboxylato ligands including their α-ether oxygen atoms, resulting in an overall two-dimensional coordination network. The thermal decomposition behavior of 4 and 5 was studied by TG-MS, revealing that decarboxylation occurs initially giving [M(CH2OC2H4OMe)2], which further decomposes by M-C, C-O and C-C bond cleavages. Complexes 4 and 5 were used as CCVD (combustion chemical vapour deposition) precursors for the deposition of Co3O4, crystalline Mn3O4 and amorphous Mn2O3 thin films on silicon and glass substrates. The deposition experiments were carried out using three different precursor solutions (0.4, 0.6 and 0.8 M) at 400 °C. Depending on the precursor concentration, particulated layers were obtained as evidenced by SEM. The layer thicknesses range from 32 to 170 nm. The rms roughness of the respective films was determined by AFM, displaying that the higher the precursor concentration, the rougher the Co3O4 surface is (17.4-43.8 nm), while the manganese oxide films are almost similar (6.2-9.8 nm).

Expectations in giving and receiving help among nurses and Russian refugees.

Stimulated by the emergence of a new refugee group that is unknown and vulnerable to misunderstanding by health care providers, and the exigency in nursing to understand potential variance in nurse and client assumptions about helping, this paper reports exploratory research comparing helping orientations of Russian Protestant Pentecostal refugees (N = 28) with professional nurses (N = 32) in the United States. Findings based on the Brickman et al. (1982) theory of helping, which addresses attribution of responsibility in helping interactions, indicated differences among the nurse and Russian orientations. Contrasting with the nurses' primary selection of the moral orientation, which assumes high individual responsibility for the causes and solutions of problems, the Russian refugee group's first selection was the medical orientation, which assumes low individual responsibility for the causes and solutions of problems. The two groups' differing orientations are considered in the context of nurse-client interactions and divergent cultural values bearing on assumptions of individual responsibility.

Anorexia as symbolic expression of a woman's rejection of her mother's life.

Studies have failed to find consistent pathologic characteristics of mothers of women with anorexia nervosa. Yet the anorexic woman's perception of her mother remains implicated in the illness (Brumberg, 1988). Literature based on anecdotal and clinical reports of anorexia nervosa has questioned anorexic women's views of their mothers and their mothers' lives. Comparison of symptomatic and asymptomatic college women found that women with anorexia nervosa described more negative impressions of their mothers' lives. Both symptomatic and asymptomatic women expressed positive feelings for their mothers, but symptomatic women tended to say their mother's lives were perhaps fulfilling for the mothers but not desirable for themselves or that their mothers' lives were very difficult and dissatisfying for the mothers and they would not want such lives for themselves. Perception of maternal role model may be related to anorexia nervosa.

Stimulation of gene expression by introns: conversion of an inhibitory intron to a stimulatory intron by alteration of the splice donor sequence.

Efficient expression of many mammalian genes depends on the presence of at least one intron. We previously showed that addition of almost any of the introns from the mouse thymidylate synthase (TS) gene to an intronless TS minigene led to a large increase in expression. However, addition of intron 4 led to a reduction in minigene expression. The goal of the present study was to determine why TS intron 4 was unable to stimulate expression. Insertion of intron 4 into an intron-dependent derivative of the ribosomal protein L32 gene did not lead to a significant increase in expression, suggesting that its inability to stimulate expression was due to sequences within the intron. Deleting most of the interior of intron 4, improving the putative branch point, removing purines from the pyrimidine stretch at the 3' end of the intron, or removing possible alternative splice acceptor or donor sites within the intron each had little effect on the level of expression. However, when the splice donor sequence of intron 4 was modified so that it was perfectly complementary to U1 snRNA, the modified intron 4 stimulated expression approximately 6-fold. When the splice donor site of TS intron 1 (a stimulatory intron) was changed to that of TS intron 4, the modified intron 1 was spliced very inefficiently and lost the ability to stimulate mRNA production. Our observations support the idea that introns can stimulate gene expression by a process that depends directly on the splicing reaction.

Introns are essential for growth-regulated expression of the mouse thymidylate synthase gene.

The thymidylate synthase (TS) gene is expressed at much higher levels in proliferating cells than in quiescent cells. We have been studying the sequences that are important for regulating the mouse TS gene. We previously showed that DNA sequences upstream of the essential promoter elements as well as downstream of the ATG codon are both necessary (but neither is sufficient) for normal regulation in growth-stimulated cells. In the present study, we examined the possible roles of the coding region, polyadenylation signal, and introns as downstream regulatory elements. Minigenes consisting of 1 kb of the TS 5'-flanking region, the coding region (with or without various introns at their normal locations), and polyadenylation signals from the TS gene, the human beta-globin gene, and the bovine growth hormone gene were stably transfected into wild-type mouse 3T6 cells. Minigenes that contained introns 5 and 6, 1 and 2, or 1 alone were regulated regardless of which polyadenylation signal was included. A minigene that contained an internally deleted version of intron 1 was also regulated in response to growth stimulation. However, when all introns were omitted, there was little if any change in the level of minigene expression as cells progressed from G1 through S phase. These observations indicate that TS introns contain sequences that are necessary for normal growth-regulated expression of the mouse TS gene. These sequences appear to be associated with sequences that are important for splicing and to function in cooperation with upstream regulatory elements to bring about normal S-phase-specific expression.