Symptoms and QOL in breast cancer patients receiving hormone therapy in Japan.

BACKGROUND: The aim was to clarify subjective symptoms and quality of life (QOL) in breast cancer patients receiving hormone therapy. METHODS: After obtaining approval from the research ethics committee, an observational study using a self-administered questionnaire was conducted at outpatient clinical oncology offices in 3 facilities that targeted breast cancer patients under the age of 50 who had been undergoing hormone therapy for less than 1 year. The study examined elements such as the breast cancer patients' basic information, symptoms, pain in daily life, QOL, and depression/anxiety. RESULTS: There were 214 valid responses. The respondents had an average age of 43.6. Of them, 100% were also treated with Tamoxifen and 30% with LH-RH agonist. There were 75% who were cognizant of side effects. Difficult symptoms that occurred with high frequency were stiff shoulders/back pain, decreased physical strength, hot flashes, and sweating. Over half the respondents were uncertain as to whether the subjective symptoms were side effects. They lost confidence in their physical strength and felt distressed over weight gain. There were 51 with a HADS anxiety score of 8 or higher, and 46 who scored 8 points or higher for depression. CONCLUSION: Breast cancer patients undergoing hormone therapy experience a variety of pains, and some also have serious psychological symptoms. Reassessing support systems to examine screening and self-care support is an issue going forward.

Developing a checksheet for breast cancer patients receiving endocrine therapy-examining reliability and validity.

BACKGROUND: The aim was to develop a checksheet that comprehends the daily lives of breast cancer patients receiving endocrine therapy, and examine its reliability and validity. METHODS: An anonymous, cross-sectional questionnaire was given to breast cancer patients under the age of 50 who had been receiving endocrine therapy for less than 1 year at outpatient clinical oncology offices in three facilities. Reliability was examined using Cronbach's alpha and test-retest reliability. Content validity, face validity, concurrent validity, convergent validity/discriminant validity, and factor analysis were used to examine validity. The study was carried out with approval from the research ethics committee. RESULTS: The checksheet was composed of three sections: subjective symptoms, discomforts in daily life while undergoing treatment, and coping with stress. Content validity was assessed as being mostly appropriate, and items identified as needing revision were corrected. Factor analysis led to four factors and 22 items for subjective symptoms, five factors and 18 items for daily life while undergoing treatment, and six factors and 24 items for coping with stress. Results indicated a high degree of correlation for concurrent validity, a moderate degree of correlation for convergent validity, and a low degree of correlation for discriminant validity. CONCLUSION: This study verified that the checksheet in its initial development as a scale has high reliability and validity.

Thrombopoietin induces production of nucleated thrombocytes from liver cells in Xenopus laevis.

The development of mammalian megakaryocytes (MKs) and platelets, which are thought to be absent in non-mammals, is primarily regulated by the thrombopoietin (TPO)/Mpl system. Although non-mammals possess nucleated thrombocytes instead of platelets, the features of nucleated thrombocyte progenitors remain to be clarified. Here, we provide the general features of TPO using Xenopus laevis TPO (xlTPO). Hepatic and splenic cells were cultured in liquid suspension with recombinant xlTPO. These cells differentiated into large, round, polyploid CD41-expressing cells and were classified as X. laevis MKs, comparable to mammalian MKs. The subsequent culture of MKs after removal of xlTPO produced mature, spindle-shaped thrombocytes that were activated by thrombin, thereby altering their morphology. XlTPO induced MKs in cultured hepatic cells for at least three weeks; however, this was not observed in splenic cells; this result demonstrates the origin of early haematopoietic progenitors in the liver rather than the spleen. Additionally, xlTPO enhanced viability of peripheral thrombocytes, indicating the xlTPO-Mpl pathway stimulates anti-apoptotic in peripheral thrombocytes. The development of thrombocytes from MKs via the TPO-Mpl system in X. laevis plays a crucial role in their development from MKs, comparable to mammalian thrombopoiesis. Thus, our results offer insight into the cellular evolution of platelets/MKs in vertebrates. (200/200).

Cellular characterization of thrombocytes in Xenopus laevis with specific monoclonal antibodies.

Platelets are produced from megakaryocytes (MKs) in the bone marrow. In contrast, most nonmammalian vertebrates have nucleated and spindle-shaped thrombocytes instead of platelets in their circulatory systems, and the presence of MKs as thrombocyte progenitors has not been verified. In developing a new animal model in adult African clawed frog (Xenopus laevis), we needed to distinguish nucleated thrombocytes and their progenitors from other blood cells, because the cellular morphology of activated thrombocytes resembles lymphocytes and other cells. We initially generated two monoclonal antibodies, T5 and T12, to X. laevis thrombocytes. Whereas T5 recognized both thrombocytes and leukocytes, T12 specifically reacted to spindle-shaped thrombocytes. The T12(+) thrombocytes displayed much higher DNA ploidy than nucleated erythrocytes, and they expressed CD41 and Fli-1. In the presence of CaCl2, adenosine diphosphate, thrombin, or various collagens, T12(+) thrombocytes exhibited aggregation. These thrombocytes were located predominantly in the hepatic sinusoids and the splenic red pulp, suggesting that both organs are the sites of thrombopoiesis. Notably, circulating thrombocytes exhibited lower DNA ploidy than hepatic thrombocytes. Intraperitoneal administration of T12 produced immune thrombocytopenia in frogs, which reached a nadir 4 days postinjection, followed by recovery, suggesting that humoral regulation maintained the number of circulating thrombocytes. Although differences between MKs and thrombocytes in X. laevis remain to be defined, our results provide further insight into MK development and thrombopoiesis in vertebrates.

Efficient approach for simultaneous estimation of multiple health-promoting effects of foods.

The investigation of new food constituents for purposes of disease prevention or health promotion is an area of increasing interest in food science. This paper proposes a new system that allows for simultaneous estimation of the multiple health-promoting effects of food constituents using informatics. The model utilizes expression data of intracellular marker proteins as descriptors that reply to stimulation of a constituent. To estimate three health-promoting effects, namely, cancer cell growth suppression activity, antiviral activity, and antioxidant stress activity, each model was constructed using expression data of marker proteins as input data and health-promoting effects as the output value. When prediction performances of three types of mathematical models constructed by simple, multiple regressions, or artificial neural network (ANN), were compared, the most adequate model was the one constructed using an ANN. There were no statistically significant differences between the actual data and estimated values calculated by the ANN models. This system was able to simultaneously estimate health-promoting effects with reasonable precision from the same expression data of marker proteins. This novel system should prove to be an interesting platform for evaluation of the health-promoting effects of food.

Dominant dystrophic epidermolysis bullosa caused by a novel G2037R mutation and by a known G2028R mutation in the type VII collagen gene (COL7A1).

An autosomal dystrophic epidermolysis bullosa (DDEB) is a hereditary mechanobullous disease characterized by blistering of the skin and the mucous membrane. DDEB is caused by a heterozygous mutation in the COL7A1 gene encoding type VII collagen, the major component of anchoring fibrils, and phenotypically classified into several types. We experienced two boys with DDEB and examined the mutation analyses of the COL7A1 genes of the two patients and their fathers to clarify the relationship between the genotypes and phenotypes, that is, the mutation sites of COL7A1 gene and the clinical types of DDEB. The case 1 and 2 patients and their fathers revealed a heterozygous nucleotide G to A transition at position 6109 and 6082 in 73 exon of COL7A1, which resulted in a glycine to arginine substitution (G2037R and G2028R), respectively. G2037R found in the case 1 patient was a novel mutation. There was no clear relationship recognized between the two mutation sites in the COL7A1 gene and the clinical variations.

Genotype analysis of Candida albicans isolates obtained from different body locations of patients with superficial candidiasis using PCRs targeting 25S rDNA and ALT repeat sequences of the RPS.

BACKGROUND: Several molecular biology-based genotyping techniques have been adapted for studying the molecular characteristics of Candida albicans strains, which constitute the majority of the etiologic agents in candidiasis. Recently, we reported a PCR system targeting 25S rDNA and ALT repeat sequences in the repetitive sequence (RPS) for genotyping of C. albicans. OBJECTIVE: To assess the potential of 25S rDNA and RPS-based genotyping for studying the molecular epidemiology of C. albicans, and define the genotypic relationship of C. albicans between invasive and non-invasive lesions in the same individual. METHODS: C. albicans strains were isolated from infected lesions and commensal sites, such as oral mucosa and/or feces, of patients with superficial candidiasis. The genomic DNAs were amplified by PCRs using P-I and P-II to determine the 25S rDNA- and RPS-based genotypes of the isolates. RESULTS: Genotype A:3 C. albicans constituted the majority of the isolates, followed by A:3/4 and B:3 C. albicans. There was usually one genotype of C. albicans per person. The genotypes of infected lesion isolates and non-infected oral mucosa and/or feces isolates were identical in the same individual, even in serially isolated C. albicans. CONCLUSION: The results indicate that our combined PCR technique using P-I and P-II is a potential tool for molecular typing of C. albicans, and reveal that the genotypes of isolates are identical in the same individual, independent of the infective and non-infective phases or the body location.

Comparison of three cytologic preparation methods and immunocytochemistries to distinguish adenocarcinoma cells from reactive mesothelial cells in serous effusion.

We assessed whether a panel of seven antibodies is useful in the differentiation of adenocarcinoma cells (ACCs) from reactive mesothelial cells (RMCs) in effusion samples and to determine optimal specimen preparation conditions for immunocytochemical analysis of effusion samples. Immunocytochemistry (ICC) was performed on three types of effusion preparations from the same effusion specimens: ethanol-fixed smears, ethanol-fixed cell-blocks, and formalin-fixed cell-blocks. Commercially available antibodies MOC-31, Ber-EP4, CA19-9, CEA, EMA, CA125, and HBME-1 were tested on RMCs from four samples of various etiology and 15 samples of adenocarcinoma from various primary sites. Ethanol-fixed smears showed strong immunoreactivity to all antibodies tested. The immunoreactivity of ethanol-fixed and formalin-fixed cell-blocks was significantly lower with all antibodies except CA19-9. Smear preparations are more sensitive than cell-blocks for immunocytochemical study. A panel of antibodies MOC-31, Ber-EP4, CA19-9, and CEA appears to be suitable to distinguish between ACCs and RMCs.

Genotyping of Candida albicans on the basis of polymorphisms of ALT repeats in the repetitive sequence (RPS).

BACKGROUND: Candida albicans is one of the most important etiologic agents causing superficial and deep fungal infections. For prevention of candidiasis, it is important to develop a rapid system that discriminates C. albicans at the strain level. OBJECTIVE: To develop a system that can identify C. albicans at the strain level. METHODS: Genomic DNAs were purified from 179 clinical isolates of C. albicans, and were used as templates for PCR amplification of 25S rDNA and ALT repeats in repetitive sequences (RPSs). PCR products generated from ALT repeats were digested with EcoRI and/or ClaI in order to study the relationships between restriction profiles and amplification profiles. RESULTS: One hundred and seventy nine clinical isolates were grouped into genotypes A (92 isolates), B (38 isolates) and C (49 isolates) on the basis of their 25S rDNA, and each was further classified into five types (types 3, 4, 3/4, 2/3/4 and 3/4/5) by PCR amplification targeting ALT repeats. Type 3 C. albicans constituted the majority of isolates in any genotypes (66.3% for genotype A, 76.3% for genotype B and 73.4% for genotype C). Each C. albicans type showed several amplification patterns, indicating the existence of subtypes. RFLP analysis revealed that restriction profiles of PCR products corresponded to amplification patterns from PCR. CONCLUSION: The present results indicate that PCR amplifications targeting 25S rDNA and ALT repeats are useful for rapid genotyping and distinction of C. albicans involved in superficial candidiasis.

Rapid identification of Candida albicans and its related species Candida stellatoidea and Candida dubliniensis by a single PCR amplification using primers specific for the repetitive sequence (RPS) of Candida albicans.

BACKGROUND: Candidiasis is caused by several Candida species, of which Candida stellatoidea and C. dubliniensis are phenotypically close to C. albicans. Although current molecular biology-based techniques can distinguish between C. albicans and C. dubliniensis, a convenient tool that can distinguish C. stellatoidea from C. albicans has not yet been developed. OBJECTIVE: To develop a system that can simply, rapidly and specifically distinguish C. albicans from the related Candida species C. stellatoidea and C. dubliniensis. MATERIALS: Genomic DNAs were purified from various yeast species and amplified by primers specific for the repetitive sequence (RPS) of C. albicans. The PCR products were purified and sequenced in order to test the specificity of the PCR amplification. RESULTS: The PCR primers only amplified several products from C. albicans, C. stellatoidea and C. dubliniensis. Sequence analysis of the products revealed that C. stellatoidea and C. dubliniensis both had RPSs including alt repeats, similar to C. albicans. After the PCR amplification, each of the three Candida species showed a unique amplification profile. Furthermore, RFLP analysis of the PCR products using EcoRI and ClaI produced species-specific restriction profiles. CONCLUSIONS: This PCR-based technique targeting the alt repeats in the RPS is useful as a tool for the rapid identification and distinction of C. albicans, C. stellatoidea and C. dubliniensis.

Mesenteric paraganglioma: report of a case.

We report a rare case of a solitary primary paraganglioma arising in the mesentery, found in a 72-year-old woman who presented with abdominal pain and a palpable abdominal mass. This extra-abdominal paraganglioma developed from paraganglionic cells that travelled by vertebral migration from the root of the superior mesenteric artery. Extra-adrenal paraganglia extend anywhere from the neck down to the base of the pelvis. Ultrasonography, computed tomography (CT), and angiography showed a solid and cystic heterogeneously enhanced mass, which was fed by the superior mesenteric artery, without distant metastasis. Exploratory laparotomy revealed a large, dark, brownish-red mass in the mesentery of the ileum, which was distinct from the ileum. The mass consisted of peripherally solid areas with central hemorrhage and cystic degeneration. It was diagnosed as a paraganglioma histologically. The patient is free from recurrence of paraganglioma after 1 year of follow up. To our knowledge, this represents only the seventh case of a paraganglioma arising in the mesentery.

Efficient secretion of human lysozyme from the yeast, Kluyveromyces lactis.

Efficient secretion of human lysozyme from the yeast, Kluyveromyces lactis, was achieved by using more stable vectors in the order of S11 replication origin-containing episomal vector < full-length K. lactis plasmid pKD1-containing vector < centromeric vector < chromosome-integrated vectors. Cells containing a PGK (phosphoglycerate kinase) promoter-driven integration vector grown in non-selective rich medium achieved the highest level of secretion, approximately 100 microg lysozyme secretion ml(-1) culture: this level was approximately 10-fold higher than that achieved by episomal vectors. An additional copy of the protein disulfide isomerase gene further facilitated the secretion.