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Background Metastasis to the breast is rare. We herein report a patient with metastatic medullary thyroid carcinoma to the breast for whom measuring the calcitonin level was an important clue to the correct diagnosis. Case presentation A 54-year-old woman visited our hospital for the treatment of recurrent metastatic medullary thyroid carcinoma due to multiple endocrine neoplasia 2A and breast cancer. Positron emission tomography performed before the operation for metastatic medullary thyroid carcinoma recurrence in the neck showed the accumulation of 18F-fluorodeoxyglucose in the bilateral breast at sites other than the disease in the neck. Ultrasonography revealed multiple tumors in both breasts. A core needle biopsy of three breast tumors was performed. Microscopically, the tumor cells showed solid growth and did not show a tubular structure. She was diagnosed with triple-negative invasive ductal carcinoma. Post-operative positron emission tomography was performed as the serum calcitonin level increased after the operation. The accumulation of 18F-fluorodeoxyglucose in the bilateral breast tumors and lymph nodes in the neck was noted. The possibility of the breast tumors being metastasis of metastatic medullary thyroid carcinoma was considered. Needle aspiration was performed for three breast tumors. The calcitonin level of the washout fluid was measured and found to be ≥ 17,500 pg/mL. Immunohistochemistry showed that the tumor cells were calcitonin-positive and gross cystic disease fluid protein-15-negative. Vandetanib was started as recurrent metastatic medullary thyroid carcinoma with breast metastasis was finally diagnosed. The serum calcitonin level decreased after 1 month. Conclusion Although breast metastasis of medullary thyroid carcinoma is rare, a correct diagnosis is indispensable for appropriate treatment. When a breast tumor shows atypical morphological features for breast cancer according to the histopathology in a patient with a history of cancer, metastasis to the breast should be considered. Calcitonin measurement of the needle washout fluid was useful for confirming metastatic medullary thyroid carcinoma.

The quality of diagnostic tests conducted by the Animal Quarantine Service (AQS) is critical to the safety of national livestock production. In procedures for tests such as polymerase chain reaction and enzyme-linked immunosorbent assay, measurement of tiny quantities of samples or reagents is quite common. Therefore, many efforts have been made to maintain the quality of micropipette operations, including routine calibration of equipment and training programs for operators. In this study, we developed a pilot training program to analyze the effects of training on pipetting accuracy, i.e., trueness and precision. The program involved a self-instruction A4-sized leaflet describing how to improve pipetting technique. Thirty-three staff members from seven AQS laboratories participated in the program. Each participant repeated sampling of 300 μL of distilled water 10 times before and after reading the leaflet, and the effects of training on the trueness and precision of the pipetting results were analyzed. To determine pipetting trueness, we examined the variation of the delivered volume from the target volume (300 μL) using the proportional odds mixture model. Our data demonstrated that participation in the training program and the level of experience with pipetting significantly improved trueness. Analysis of standard deviations of the ten samplings, as an indicator of precision, also showed that participation in the training program improves pipetting precision. Thus, our data supported the implementation of such simple, quick training sessions to improve pipetting trueness and precision in diagnostic laboratories.

Mounting evidence suggests that glutamate excitotoxicity induces both enzymatic cleavage and nuclear translocation of apoptosis-inducing factor (AIF), which is involved in apoptosis-like programed cell death characterized by nuclear condensation without appearance of apoptotic bodies. Given the lack of apoptotic bodies in motor neurons in the spinal cord of patients with amyotrophic lateral sclerosis (ALS), the aim of the present study was to determine the role for AIF in this disease. We investigated the expression of AIF in spinal cords obtained at autopsy from ten sporadic ALS patients and ten age-matched, control subjects, using morphological and quantitative techniques. Immunohistochemical analysis showed that AIF immunoreactivity was localized in the nucleus as well as the cytoplasm of a subset of affected motor neurons and reactive astrocytes in the ALS cases, while it was restricted to the cytoplasm of these cells in the control cases. Immunoblot analysis disclosed immunoreactivity for cleaved AIF in both cytoplasmic and nuclear protein extracts at a 57-kDa mobility. Densitometric analysis revealed significant increases in the cytoplasmic cleaved AIF/cytoplasmic β-actin ratio and the nuclear cleaved AIF/nuclear histone H1 ratio in the ALS group compared with the control group. There was no significant link between the cytoplasmic and nuclear cleaved AIF levels in the ALS spinal cords and the clinical features such as phenotypes, age at death, and disease duration. Our results provide evidence for persistent cleavage and nuclear translocation of AIF in ALS spinal cord, suggesting implications for the AIF-mediated motor neuron death in this disease.

Treating patients with severe aplastic anemia (SAA) who fail to respond to immunosuppressive therapy (IST) and do not have an HLA-matched donor is challenging. We report favorable outcomes in 11 patients who underwent bone marrow transplantation (BMT) from an HLA-mismatched unrelated donor. The median age was 11 years (range, 3-20 years). The conditioning regimen consisted of cyclophosphamide (200 mg/kg), antithymocyte globulin (10 mg/kg), and total body irradiation (5 Gy). Patients received tacrolimus and methotrexate for prophylaxis against graft-versus-host disease (GVHD). Donorrecipient pairs were mismatched for the HLA-DR antigen in 8 patients by serologic typing. HLA-A and HLA-B antigens were mismatched in 1 and 2 patients, respectively. Ten patients achieved engraftment. One patient who failed to engraft was rescued by a second transplantation from her mother, who was mismatched at 2 HLA antigens. Acute GVHD of grades II to IV occurred in 2 patients. Three patients developed limited chronic GVHD, and 1 patient developed extensive chronic GVHD of the lung. All patients are alive at 9 to 56 months after transplantation (median, 33 months). Considering our encouraging results, HLA-mismatched unrelated-donor BMT for SAA is feasible as a salvage therapy for nonresponders to IST.

Several studies have documented the involvement of oxidative stress represented by lipid peroxidation in the pathogenesis of Alzheimer's disease (AD). To test whether the highly reactive carbonyl crotonaldehyde (CRA), generated during lipid peroxidation, is involved in AD, we performed an immunohistochemical analysis in AD and age-matched control hippocampi using a specific antibody against protein-bound CRA (P-CRA). In the AD cases, P-CRA immunoreactivity was preferentially localized in reactive astrocytes and microglia around senile plaques (SPs) and those present in the neuropil, while it was weakly detectable in neurons and neurofibrillary tangles. P-CRA immunoreactivity was also localized in all portions of diffuse SPs and the dystrophic neurites of neuritic and classical SPs, but was undetectable in amyloid cores. Age-matched controls showed P-CRA immunoreactivity only very weakly in neurons. In contrast to P-CRA, immunoreactivities for protein-bound acrolein and 4-hydroxy-2-nonenal were mainly localized to neurons and rarely seen in glial cells. Our results suggest that increased oxidative stress and CRA formation in glial cells is implicated in the disease processes of AD.

Recent studies have documented carbonyl stress involvement in the pathogenesis of sporadic amyotrophic lateral sclerosis (ALS). The aim of the present study was to assess a role for carbonyl stress in motor neuron degeneration associated with superoxide dismutase-1 (SOD1) mutant familial ALS and its transgenic mouse model, using an immunohistochemical investigation of advanced glycation end products (AGEs) and advanced lipoxidation end products (ALEs). In the spinal cords from six familial ALS patients with SOD1 A4V mutation and six transgenic mice expressing G93A mutant human SOD1, immunoreactivities for N(epsilon)-(carboxyethyl)lysine, argpyrimidine, pyrraline and N(epsilon)-(carboxymethyl)lysine as AGEs were distinct in almost all of the reactive astrocytes and obscure in the residual neurons, whereas no immunoreactivity for pentosidine as an AGE, or 4-hydroxy-2-nonenal-histidine, malondialdehyde-lysine or acrolein-lysine as ALEs was detectable. Spinal cords from age-matched control humans and mice exhibited no significant immunoreactivities for the examined products. Our results indicate that protein glycation, but not lipid peroxidation, is enhanced in ALS patients with an SOD1 mutation and mutant SOD1 transgenic mice, in which certain AGEs are selectively formed in the spinal cord astrocytes.

This report describes a case of aortic papillary fibroelastoma causing transient ischemic attack (TIA), and is followed by a review of the relevant literature. A tumor measuring 1.5 x 1.5 cm was located in the right coronary cusp of the aortic valve, which was found to have a "sea anemone-like" configuration due to collagenous or elastic papillary projections. Aortic fibroelastoma can cause cerebrovascular symptoms, and this patient presented with TIA. Moreover, aortic fibroelastoma usually causes chest symptoms and occasionally results in death despite its small size of generally less than 1 cm. According to the literature, all of the patients who died of fibroelastoma had an aortic valve tumor in the right or left coronary cusp, which indicates that the location of the tumor rather than its size may be an important factor contributing to the case of death. Surgical treatment is usually indicated especially for aortic fibroelastoma because of the high associated risk or cerebrovascular and chest diseases. The recent evolution of echocardiography will promote the chance of establishing a preoperative diagnosis of this lesion. However, because it is not possible to differentiate fibroelastoma from other lesions, including malignancies, by echocardiography alone, this may be another reason for performing surgical removal.

The adhesion of Vibrio cholerae O1 strains to human intestinal epithelial cell, Intestine 407, was analyzed by flow cytometer. According to positive percentages of Intestine 407 cells adhered by V. cholerae, two groups of V. cholerae strains were classified as follows: more adhesive (more than 50%), less adhesive (less than 50%) strains. In addition, the fluorescence intensity after attachment of V. cholerae was directly correlated to the number of the microorganisms. It was concluded that flow cytometry is a useful and objective method for analyzing adhesion of V. cholerae to cultured cells.

Background Emerging evidence suggests that innate immunity and increased oxidative stress contribute to pathomechanisms in amyotrophic lateral sclerosis (ALS). The aim of the present study was to verify the involvement of monocyte chemoattractant protein-1 (MCP-1) and its specific CC chemokine receptor 2 (CCR2) in the disease progression of ALS. We here demonstrate the expression state of MCP-1 and CCR2 in lumbar spinal cords of mice overexpressing a transgene for G93A mutant human superoxide dismutase 1 (SOD1) (ALS mice) as a mouse model of ALS as well as the involvement of MCP-1/CCR2-mediated signaling in behavior of cultured astrocytes derived from those mice. Results Quantitative polymerase chain reaction analysis revealed that MCP-1 and CCR2 mRNA levels were significantly higher in ALS mice than those in nontransgenic littermates (control mice) at the presymptomatic stage. Immunoblot analysis disclosed a significantly higher CCR2/β-actin optical density ratio in the postsymptomatic ALS mouse group than those in the age-matched control mouse group. Immunohistochemically, MCP-1 determinants were mainly localized in motor neurons, while CCR2 determinants were exclusively localized in reactive astrocytes. Primary cultures of astrocytes derived from ALS mice showed a significant increase in proliferation activity under recombinant murine MCP-1 stimuli as compared to those from control mice. Conclusions Our results provide in vivo and in vitro evidence that MCP-1 stimulates astrocytes via CCR2 to induce astrocytosis in ALS with SOD1 gene mutation. Thus, it is likely that MCP-1/CCR2-mediated sigaling is involved in the disease progression of ALS.

DNA sensitive field-effect transistors (DNA-FET) have been realized using single crystalline diamond grown by plasma-enhanced chemical vapor deposition (CVD). To bond DNA to diamond, amine linker-molecules are covalently attached by photochemical means to H-terminated diamond surfaces. Using hetero-bifunctional cross-linker and thiol-modified single-strand (ss) marker DNA, the gate of diamond FETs is modified to sense hybridization of DNA, forming double-strand (ds) DNA molecules on the gate. The density of DNA bonded to diamond is varied between 1012 and 1013 cm−2 to explore sensitivity enhancements by reduction of the DNA molecule density. DNA-FET characterization in 1M NaCl buffer solution (pH 7.2) reveal gate-potential threshold shifts by hybridization in the range 30 mV to 100 mV with decreasing DNA density. The variation is discussed based on the transfer doping model which predicts with decreasing pH increasing hole-densities in the surface conductive layer of diamond.

This book is a manual for the zoological study of intertidal invertebrates. This manual was originally created in 1941 as a class syllabus for undergraduate students in invertebrate zoology. Now in the 4th edition, the number of authors has increased from 22 (2nd edition, 1954) to 120 and the study area has been expanded as well, covering Point Conception to the Oregon–Washington border. The improvements, including newly added information, in this manual serve as a good guide not only for students but also for professional natural scientists. This manual is composed of two parts. The first part, with six chapters, is a general introduction to field study and intertidal invertebrate animals. It deals with the environmental characteristics of intertidal habitats and their effects on the life of invertebrates; biogeography in this area; survey and processing methods of each group of invertebrates; and recent topics such as molecular identification and introduced species. It is remarkable that every chapter has specific examples of studies conducted in the study area dealing not only with environmental characteristics of each habitat or biogeography, but also with molecular studies. The second part consists of taxonomic accounts introducing the taxonomical position of each phylum, morphological keys for identification of species, genus, family or subclass, and species list. It introduces 31 phyla from Protista to Chordata. This part includes fine illustrations or black and white photographs of the species collected from the study area. They illustrate their lives in the field effectively. Each chapter also explains the basic body structure, ecology, and techniques for studies such as sampling, treatment, and identification of the phylum. It is notable that much valuable information is accumulated in a subset of the Pacific Coast of North America. These taxonomic accounts will assist not only beginners in this field but also zoologists who work on invertebrate animals in other areas of the world. For community ecologists working on intertidal benthic organisms, the study area represents a special one because it is where most contributive studies for setting up the principle of community organization were conducted. Menge, Lubchenco, Sutherland, and many other ecologists contributed experimental studies to figure out the determining factor of community structure on the coast of Oregon. They proved that the predators that forage on the dominant species in the prey community have a strong influence on the community because they can moderate the competition for space and some of them can determine their vertical distribution. This effect is known as the top– down effect. These ecologists also stressed that the strength and direction of the top–down effect is changed by environmental conditions. Roughgarden brought forward supply-side ecology based on the studies conducted in the area between Point Conception and San Francisco. He pointed out that the structure of the intertidal benthic community is organized not only by environmental factors and species interactions at the adult benthic stage but also by the supply of larvae. Since the supply of planktonic larvae is governed by conditions and events in the offshore waters, his model coordinated ecology and oceanography. By reading this manual, we can clearly understand that these many T. Yamamoto (&) Faculty of Fisheries, Kagoshima University, 4-50-20 Shimoarata, Kagoshima 890-0056, Japan e-mail: yamamoto@fish.kagoshima-u.ac.jp

Background It is well known that expression of certain bacterial genes responds rapidly to such stimuli as exposure to toxic chemicals and physical agents. It is generally believed that the proteins encoded in these genes are important for successful survival of the organism under the hostile conditions. Analogously, the proteins induced in bacterial cells exposed to antibiotics are believed to affect the organisms' susceptibility to these agents.

The chromosomal gene(s) for cephalosporinase production of Citrobacter freundii GN346 has been cloned into vector plasmid pMK1, initially as a 7.3 kb EcoRI fragment. From the substrate profile and the response to anti-GN346 CSase serum of the enzyme produced, it was confirmed that the hybrid plasmid (pTY71) carries the relevant chromosomal cephalosporinase gene from C. freundii GN346. A restriction endonuclease cleavage map of cloned EcoRI fragments was constructed, and the structural gene of the cephalosporinase could be limited in the 1.5 kb BamHI fragment. The cloned gene(s) was expressed at an extremely low level in Escherichia coli. Furthermore, its expression was constitutive in E. coli, although inducible in its own cytoplasm.

The fine structure of the genes tnpA, tnpR and res of Tn2603 required for its own transposition, was determined. The order of the genes was tnpA-tnpR-res from the right end of the right hand side region in Tn2603, the tnpA and tnpR encoded gene products having molecular weights of 110,000 and 21,000, respectively. The 110,000 molecular weight polypeptides was absolutely required for replicon fusion as the first stage of transposition, and named transposase. On the other hand, the 21,000 molecular weight polypeptide was necessary for resolution of the cointegrate as the second stage of transposition, and named resolvase. We also examined the ability of various transposons, assumed to be closely related, to complement the tnpA and tnpR mutations of Tn2603. The results indicated that the mercury resistance transposon, Tn2613, and Tn501, can complement both genes, but TnAs and gamma delta cannot at all. Tn501 had much less efficiency of complementation for tnpA than Tn2613. We have also discovered that the transposition frequency of transposons in the tn2613 family systematically depend on their size of transposon.