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An efficient method for the one-pot synthesis of tetramic acid derivatives was developed utilizing tandem umpolung N-alkylation/reduction/cyclization of γ-hydrazono β-ketoester. By using this reaction as a key step, the total synthesis of the 3-spiro 7-hydroxamic acid tetralin which possesses an HDAC inhibitory activity was also achieved.

Summary To better understand the association between high salt intake and osteoporosis, we investigated the effect of sodium chloride (NaCl) on mice and human osteoclastogenesis. The results suggest a direct, activating role of NaCl supplementation on bone resorption. Introduction High NaCl intake is associated with increased urinary calcium elimination and parathyroid hormone (PTH) secretion which in turn stimulates the release of calcium from the bone, resulting in increased bone resorption. However, while calciuria after NaCl loading could be shown repeatedly, several studies failed to reveal a significant increase in PTH in response to a high-sodium diet. Another possible explanation that we investigated here could be a direct effect of high-sodium concentration on bone resorption. Methods Mouse bone marrow macrophage and human peripheral blood mononuclear cells (PBMC) driven towards an osteoclastogenesis pathway were cultivated under culture conditions mimicking hypernatremia environments. Results In this study, a direct effect of increased NaCl concentrations on mouse osteoclast differentiation and function was observed. Surprisingly, in a human osteoclast culture system, significant increases in the number of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts, calcitonin receptor (CTR)-positive osteoclasts, nuclear factor-activated T cells c1 (NFATc1) gene expression, and areal and volumetric resorptions were observed for increasing concentrations of NaCl. This suggests a direct, activating, cell-mediated effect of increased concentrations of NaCl on osteoclasts. Conclusions The reported that enhanced bone resorption after high-sodium diets may not only be secondary to the urinary calcium loss but may also be a direct, cell-mediated effect on osteoclastic resorption. These findings allow us to suggest an explanation for the clinical findings independent of a PTH-mediated regulation.

Wheat (Triticum aestivum L.) and rice (Oryza sativa L.), which belong to the Poaceae family, are starch-grain crops, but wheat is adapted to cooler temperature conditions than rice. In this study, the difference between the two contrasting crops in grain-filling adaptability in response to temperature was investigated. Two spring wheat cultivars were grown in the Mediterranean-type environments of Western Australia and Southeast Turkey, and four temperate-zone rice cultivars were grown in several locations in Japan under irrigated conditions. Portions of the crops were enclosed under a plastic canopy to elevate the temperature after anthesis. Average temperatures during grain filling ranged from 14 to 24 °C for wheat and from 23 to 29 °C for rice. Grain yield varied from 280 to 599 g m-2 in wheat and 354 to 736 g m-2 in rice. When plant density was halved at flowering to estimate the potential grain-filling rate under an increased supply of assimilates, the grain-filling percentages [%F, observed grain weight (G)/potential grain weight (PG)] of both crops were represented by similar logistic curves of cumulative temperatures during the grain filling period. These results suggest that grain-filling responses to temperature scarcely differ between spring wheat and temperate-zone rice. G was estimated for the spring wheat and temperate-zone rice cultivars under different temperatures after anthesis using an assimilate-limited grain-potentiality model consisting of the following parameters: rate of whole-plant weight increase (?W/?t), rate of potential grain dry weight increase (?PG/?t) based on rate of%F (%F/?t) and PG, and the amount of stem reserves (SP). The observed data showed that the decrease in ?W/?t with an increase in temperature in wheat was greater than in rice. According to the model, G started to decrease at lower temperatures in wheat than in rice, and this decrease was accelerated by lower amounts of SP. Therefore, the difference in the optimal temperatures for G during grain filling between the two crops was suggested to mainly result from the sensitivity of assimilation to high temperatures. © 2017 Elsevier B.V. Commonwealth Scientific and Industrial Research Organisation Research Institute for Humanity and Nature Japan Society for the Promotion of Science Asthma Foundation of Western Australia Our thanks go to a project in Çukurova Unviersity, Turkey of the Impact of Climate Changes on Agricultural Production System in Arid Areas (ICCAP) in the Research Institute for Humanity and Nature, Japan, and CSIRO, Western Australia for the substantial support of, and contributions to, our research. We would like to thank Dr. Neil C. Turner, University of Western Australia for his comments and suggestions to our manuscript. We thank JSPS for their financial support of the project in Australia and Japan. Appendix A

We demonstrate that disulfide bond replacement is an efficient strategy for engineering therapeutic peptides. In previous work, short peptide fragments, known as WP9QY, with sequence homology with the predicted ligand contact surface of the receptor activator of NF-κB (RANK) were crosslinked through intramolecular disulfide bonds to inhibit RANK ligand (RANKL)-induced signaling, osteoclastogenesis, bone resorption in vitro, and bone loss in vivo. We report that replacement of the disulfide bond of WP9QY with an amine cross-linkage results in a significant improvement in enzymatic stability, with only a slight loss of bone resorption-blocking activity in vitro. Furthermore, the WP9QY derivative inhibits bone loss significantly in vivo, whereas the native form of WP9QY was not effective under the same conditions.

Mucosa-associated lymphoid tissue (MALT) lymphoma is a low-grade extra nodal often gastric condition. MALT lymphoma of the head and neck has been reported in the salivary gland, thyroid, larynx, orbit, and skin. A 76-year-old woman seen for double vision was found in neuro opthalmological examination to have trochlear and oculomoter nerve palsy. Epipharyngeal endoscopy showed a submucosal tumor in the posterior epipharynx wall. Computed tomography (CT) and magnetic resonance imaging (MRI) demonstrated a mass lesion of the epipharynx, pterygopalatine fossa, and cavernous sinus. Endoscopic tumor biopsy performed twice for the epipharynx and for the pterygopalatine fossa was pathologically difficult to differentiate from lymphoid tissue and lymphoma hypertrophy. Clonal Southern blot analysis showed clonal rearrangement of the heavy immunoglobulin gene chain, yielding a diagnosis of MALT lymphoma.

AIM The morbidity of hypertension increases with aging although the exact relationship between hypertension and menopause has not been clearly elucidated. Therefore we set out to clarify the effects of aging and menopause upon women's vascular systems. METHODS We divided 151 elderly and middle-aged women into 3 groups (premenopause group, menopause group and post-menopause group). We measured height, weight, blood pressure (BP), total cholesterol (T-chol), HDL-cholesterol (HDL-chol), LDL-cholesterol (LDL-chol), high sensitivity C-reactive protein (hsCRP), creatinine (Cr), triglyceride (TG), fasting blood glucose (FPG), IRI, HOMA-R, brachial-ankle pulse wave velocity (baPWV), augmentation index (AI), percentage of flow-mediated dilatation (%FMD), and echocardiography. RESULTS In the post-menopause and menopause groups systolic BP and AI were significantly higher than those in the pre-menopause group. There was a significant difference in systolic BP between the post-menopause group and menopause group. In the post-menopause group, baPWV, Cr, and hsCRP were significantly higher than those in the pre-menopause group. There was significant difference in baPWV between the post-menopause group and menopause group. In the post-menopause group, %FMD and eGFR were significantly lower than those in other 2 groups. In the post-menopause group and the menopause group, E/A was significantly lower than in the pre-menopause group. There was also a significant difference in E/A between the post-menopause group and the menopause group. CONCLUSIONS Elevated blood pressure, atherosclerosis and endothelial dysfunction were associated with aging and menopause in their present study. These results suggest that understanding women's cardiovascular changes which accompany aging are important for women's health care and prevention of cardiovascular events.

Endoscopic examination, including narrow-band imaging (NBI), is being increasingly used to detect early hypopharygeal cancer. Because these high-risk cases have already often undergone radiotherapy for esophageal cancer, early hypopharygeal cancer is a good indication for endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD). Slightly larger tumors, however, require partial hypopharyngotomy preserving the larynx. Since reconstruction with a free flap may be highly invasive, we reported using an anterior chest skin flap as a useful alternative in pharyngeal reconstruction.

Pediatric thyroid gland carcinoma is uncommon, representing less than 1% of all thyroid cancer. We report cases of thyroid carcinoma in a 9-year-old boy and a 12-year-old girl, both with dyspnea and lateral cervical tumors, and both found in computed tomography (CT) and fine-needle aspiration cytology. Lung metastasis was found in chest CT on presentation. Because both were in the bilateral cervix, we conducted total thyroidectomy and bilateral selective neck dissection level 2-6. Case 1 had recurrent laryngeal nerve and tracheal invasion. Radioactive iodine therapy was administered to both postperatively for lung metastasis. Children with well-differentiated papillary and follicular carcinoma are reported to have a substantially better prognosis than adults despite comparatively high regional and distant metastasis incidence. Appropriate diagnosis, surgical procedure choice, and careful follow-up are thus important in pediatric thyroid cancer.

Formation of osteoclasts and consequent joint destruction are hallmarks of rheumatoid arthritis (RA). Here we show that LIGHT, a member of the tumour necrosis factor (TNF) superfamily, induced the differentiation into tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells (MNCs) of CD14(+) monocytes cocultured with nurse-like cells isolated from RA synovium, but not of freshly isolated CD14(+) monocytes. Receptor activator of nuclear factor-kappaB ligand (RANKL) enhanced this LIGHT-induced generation of TRAP-positive MNCs. The MNCs showed the phenotypical and functional characteristics of osteoclasts; they showed the expression of osteoclast markers such as cathepsin K, actin-ring formation, and the ability to resorb bone. Moreover, the MNCs expressed both matrix metalloproteinase 9 (MMP-9) and MMP-12, but the latter was not expressed in osteoclasts induced from CD14(+) monocytes by RANKL. Immunohistochemical analysis showed that the MMP-12-producing MNCs were present in the erosive areas of joints in RA, but not in the affected joints of osteoarthritic patients. These findings suggested that LIGHT might be involved in the progression of inflammatory bone destruction in RA, and that osteoclast progenitors might become competent for LIGHT-mediated osteoclastogenesis via interactions with synoviocyte-like nurse-like cells.

獨協医科大学越谷病院に勤務する教職員を対象に,平成18年度健康診断受診率および健康診断結果を全国平均と比較し,現状と今後の課題について検討した.胸部レントゲン検査,血圧,尿検査,心電図検査,聴力検査では要精検率が全国平均を下回ったが,男性では血中脂質,女性では貧血の項目で全国平均を上回る者が多くみられた.その要因としては,運動不足や食生活などの生活習慣の問題があると考えられ,それらの改善が必要であると推測された., To clarify actual and near future health concerns, a study was performed on the faculty and staff of Dokkyo Medical University, Koshigaya Hospital, Japan, in 2006. Chest x-ray, blood pressure, proteinuria and urine sugar, ECG (electrocardiogram), and aural test results indicated a lower percentage of abnormality when compared with the general population of Japan. A high level of hyperlipidemia in males and anemia in females was recognized in the data, especially for those past the age of fifty. It can be speculated that a change in life style is necessary, for example improvement of our dietary habits and making up for the lack of proper exercise, in order to maintain good health.

Introduction We designed OP3-4 (YCEIEFCYLIR), a cyclic peptide, to mimic the soluble osteoprotegerin (OPG), and was proven to bind to RANKL (receptor activator of NF-κB ligand), thereby inhibiting osteoclastogenesis. We recently found that another RANKL binding peptide, W9, could accelerate bone formation by affecting RANKL signaling in osteoblasts. We herein demonstrate the effects of OP3-4 on bone formation and bone loss in a murine model of rheumatoid arthritis. Methods Twenty-four seven-week-old male DBA/1J mice were used to generate a murine model of collagen-induced arthritis (CIA). Then, vehicle or OP3-4 (9 mg/kg/day or 18 mg/kg/day) was subcutaneously infused using infusion pumps for three weeks beginning seven days after the second immunization. The arthritis score was assessed, and the mice were sacrificed on day 49. Thereafter, radiographic, histological and biochemical analyses were performed. Results The OP3-4 treatment did not significantly inhibit the CIA-induced arthritis, but limited bone loss. Micro-CT images and quantitative measurements of the bone mineral density revealed that 18 mg/kg/day OP3-4 prevented the CIA-induced bone loss at both articular and periarticular sites of tibiae. As expected, OP3-4 significantly reduced the CIA-induced serum CTX levels, a marker of bone resorption. Interestingly, the bone histomorphometric analyses using undecalcified sections showed that OP3-4 prevented the CIA-induced reduction of bone formation-related parameters at the periarticular sites. Conclusion The peptide that mimicked OPG prevented inflammatory bone loss by inhibiting bone resorption and stimulating bone formation. It could therefore be a useful template for the development of small molecule drugs for inflammatory bone loss. Electronic supplementary material The online version of this article (doi:10.1186/s13075-015-0753-8) contains supplementary material, which is available to authorized users.

Objectives. Nurse-like stromal cells (NLC) in synovia and bone marrow of patients with rheumatoid arthritis (RA) can support pseudoemperipolesis, protect from apoptosis and enhance immunoglobulin production of peripheral blood B cells isolated from healthy individuals, suggesting the profound contribution of hyperactivation of B cells in RA. In the course of establishing RA-NLC from RA patients, we observed the growth of B cells in the presence of RA-NLC. Methods. We cloned B cells from the synovium or bone marrow of RA patients using the limiting dilution technique. For established clones, nucleotide sequences of immunoglobulin and surface antigens were investigated. To investigate the dependence of these clones on NLC, differences in the proliferation and the amount of immunoglobulin produced in the presence or absence of NLC were compared. Immunocytochemical staining of various cells was performed using the antibody these clones produced. Results. Nine B-cell clones established from RA patients showed RA-NLC-dependent growth. These B-cell clones expressed CD19, CD20, CD38, CD39 and CD40, suggesting that the cloned cells were mature and activated. All clones secreted immunoglobulins in culture media, which were specific for intracellular components of various cell lines, including RA-NLC. Interestingly, we found limited usage of immunoglobulin heavy-chain variable regions (VH) among B-cell clones from RA patients. These repertoires were reported to be detected preferentially in fetal livers. Conclusion. The present study provides a novel insight into the involvement of RA-NLC in the immunopathogenesis of RA via an autoreactive B cell development and/or activation mechanism.

Doc2alpha is a synaptic vesicle-associated Ca2 + -binding protein. To study the role of Doc2alpha in synaptic transmission and modulation, we generated homozygous null Doc2alpha mutant mice. In the CA1 region of hippocampal slices in the mutant mice, excitatory synaptic responses evoked with prolonged 5 Hz stimulation showed a significantly larger frequency facilitation followed by a steeper depression than those in wild-type mice, whereas there was no difference in synaptic transmission at lower frequencies or in paired-pulse facilitation. These results suggest that Doc2alpha regulates synaptic transmission when high Ca2 + concentrations in the presynaptic terminal are sustained. Furthermore, the mutant mice showed impairment in long-term potentiation and passive avoidance task. Thus, Doc2alpha may regulate transmitter release during repetitive synaptic activation, thereby contributing to memory formation.

Doc2 has one Munc13-interacting domain at the N-terminal region and two C2-like domains interacting with Ca2+ and phospholipid at the C-terminal region. Doc2 consists of two isoforms, Doc2α and -β. Doc2α is specifically expressed in neuronal cells and implicated in Ca2+-dependent neurotransmitter release, whereas Doc2β is ubiquitously expressed and its function is unknown. We show here that both Doc2α and -β interact with rat tctex-1, a light chain of cytoplasmic dynein, in both cell-free and intact cell systems. Overexpression of the N-terminal fragment of Doc2 containing the tctex-1-interacting domain induces changes in the intracellular localization of cation-independent mannose 6-phosphate receptor and its ligand, cathepsin D, which are transported from trans-Golgi network to late endosomes. Overexpression of the C-terminal fragment containing two C2-like domains shows the similar effect, but to a lesser extent, whereas overexpression of full-length Doc2 or the C-terminal fragment of rabphilin3 containing two C2-like domains does not show this effect. Because dynein is a minus-end-directed microtubule-based motor protein, these results suggest that Doc2, especially Doc2β, plays a role in dynein-dependent intracellular vesicle transport.

Munc13 is a component of the neurotransmitter release machinery which is specifically expressed in brain. Munc13 interacts with Doc2 and syntaxin which are also implicated in the neurotransmitter release process. Here we isolated another Munc13-interacting molecule from a rat brain cDNA library by use of the yeast two-hybrid system, identified it to be a novel type of beta spectrin, and named it beta SpIII sigma 1. beta SpIII sigma 1 was specifically expressed in brain, where it was enriched in the synaptic vesicle and plasma membrane fractions. Because spectrin has been shown to interact with the actin cytoskeleton which is involved in the exocytotic process, the present results suggest that the Munc13-beta SpIII sigma 1 interactions play a role in neurotransmitter release.

Histoid leprosy is a rare multibacillary form that presents with disseminated papule-nodular cutaneous lesions. To study the inflammatory infiltrate of the histoid form and to compare it with other lepromatous forms, we performed histological and immunohistochemical analysis on skin biopsies. Fifteen patients were included for histopathological analysis (10 histoid and five lepromatous) via the haematoxylin-eosin and Ziehl-Neelsen-Faraco stains. Thus, immunohistochemical techniques using immunoperoxidase assay were performed for: anti-BCG, anti-M. leprae, anti-CD8, anti-CD3, anti-CD20, anti-S100, anti-CD1a, anti-CD68 and antivimentin. Spindle cells were present in all histoid patients. A pseudocapsule was observed in half of both studied forms. A comparison using the Ziehl-Neelsen-Faraco stain to evaluate anti-BCG and anti-M.leprae showed no major differences. The CD3+ cells were more pronounced in the histoid form than the lepromatous form. There was greater immunoreactivity toward CD8+ cells in the histoid form, as well as the CD20+ cell count. A similar count of S100+ cells in the epidermis of both leprosy forms was observed. There was a slight increase of dendritic cells in the histoid patients in the superficial and deep dermis. For CD1a marker, we observed expression in the epidermis and superficial dermis in both forms. A diffuse and intense infiltrate of CD68+ cells was also observed in the histoid and lepromatous forms. The high positivity for vimentin did not allow for a positive cell count. We concluded that the activation of both the cellular and humoral response is more pronounced in the histoid form because the T and B cells showed greater infiltration than those in the lepromatous form. The activation of dendritic and Langerhans cells is similar in both forms. The spindle cells likely belong to the macrophage population, thus maintaining phagocytic ability. The quantities of pseudocapsules and bacilli are similar and cannot serve as criteria for diagnosis.

We have previously isolated from a human brain cDNA library, a new protein having two C2-like domains which interact with Ca2+ and phospholipid, and named Doc2alpha. Doc2alpha is abundantly expressed in brain, where it is highly concentrated on the synaptic vesicle fraction, and is implicated in Ca2(+)-dependent exocytosis. We have isolated here a mouse Doc2alpha cDNA and determined the localization of its mRNA in adult mouse brain. The amino acid sequence of the mouse Doc2alpha cDNA is 92% identical with that of the human counterpart. Northern blot analysis and in situ hybridization on adult mouse brain sections have revealed that Doc2alpha is predominantly expressed in mouse brain, where it is expressed in neuronal cells, but not in non-neuronal cells. Doc2alpha is highly expressed in the olfactory bulb, cerebral cortex, hippocampus, amygdaloid complex, and ventromedial hypothalamus nucleus, but not in the cerebellum, caudate-putamen, or ventral thalamus. These results indicate that Doc2alpha is expressed heterogeneously in mouse brain, where it is predominantly expressed in neuronal cells, and suggest that Doc2alpha plays a specific role in the area where it is expressed.

We previously isolated a new protein having two C2-like domains which interacted with Ca2+ and phospholipid and named Doc2 (Double C2). Because Doc2 was abundantly expressed in brain where it was highly concentrated on the synaptic vesicle fraction, we have examined here whether Doc2 is involved in Ca2+-dependent exocytosis from cultured PC12 cells. For this purpose, we took advantage of the growth hormone (GH) co-expression assay system of PC12 cells in which GH is stored in dense core vesicles and released in response to high K+ in an extracellular Ca2+-dependent manner. Northern and Western blot analyses indicated that Doc2 is present in PC12 cells. Overexpression of hemagglutinin-tagged Doc2 stimulated the Ca2+-dependent, high K+-induced release of co-expressed GH without affecting the basal release. In the PC12 cells transfected with a plasmid with the coding sequence of Doc2 in the antisense orientation, the high K+-induced release of co-expressed GH was inversely inhibited. The Doc2 mutant expressing an N-terminal fragment or a C-terminal fragment containing two C2-like domains inhibited the high K+-induced release of co-expressed GH. These results indicate that Doc2 enhances Ca2+-dependent exocytosis of dense core vesicles from PC12 cells.

Rabphilin-3A, a putative target molecule of Rab3A small GTP-binding protein implicated in Ca2+-dependent exocytosis, consists of two functionally different domains: the N-terminal Rab3A-binding domain and the C-terminal two C2-like domains (C2A and C2B domains) interacting with Ca2+ and phospholipid. Here, we used the growth hormone (GH) co-expression assay system of PC12 cells in which expressed GH is released in response to high K+. Reduction of endogenous rabphilin-3A inhibited the Ca2+-dependent, high K+-induced GH release. Various rabphilin-3A mutants expressing an N-terminal, C-terminal, or C2B fragment, but not the rabphilin-3A mutant expressing a C2A fragment, inhibited the high K+-induced GH release. These results indicate that rabphilin-3A is involved at least in Ca2+-dependent exocytosis from PC12 cells and that the C2A and C2B domains have different functions.

We previously isolated a novel protein having two C2-like domains known to interact with Ca2+ and phospholipid, and named Doc2 (Double C2). Doc2 is predominantly expressed in brain and is implicated in Ca(2+)-dependent neurotransmitter release. We have isolated here an isoform of Doc2 and named the original one Doc2 alpha and the new one Doc2 beta. Doc2 beta alsp has two C2-like domains and is 61% identical to Doc2 alpha at the amino acid level. In contrast to Doc2 alpha, the Doc2 beta mRNA is expressed ubiquitously. These results indicate that there are at least two isoforms of Doc2, and suggest that Doc2 beta is involved in Ca(2+)-dependent intracellular vesicle trafficking in various types of cells.

Objective. To investigate the morphology and function of multinucleated bone-resorbing giant cells derived from CD14-positive cells in the synovial fluids (SF) of patients with rheumatoid arthritis (RA) or osteoarthritis (OA). Methods. CD14-positive cells were obtained by magnetic-activated cell sorting of primary cultures of mononuclear cells from the SF. Multinucleated bone-resorbing giant cells were induced from the CD14-positive cells in the presence or absence of cytokines. We examined various characteristics, including osteoclast markers, fusion index and bone-resorption activities of the multinucleated giant cells. Results. Multinucleated giant cells were induced from the CD14-positive cells in the SF of the RA and OA patients by the addition of interleukin (IL)-3, IL-5 and IL-7, or granulocyte–macrophage colony-stimulating factor (GM-CSF), respectively. These multinucleated giant cells were positive for tartrate-resistant acid phosphatase (TRAP), carbonic anhydrase II, actin, vitronectin receptor and the calcitonin receptor. However, the average values for the number of nuclei, fusion index and boneresorption functions of the SF cells from the RA patients were significantly higher than those derived from the OA patients. Conclusion. These results suggest that the induction and activities of multinucleated bone-resorbing giant cells may play a pivotal role in bone destruction, and that these processes may be enhanced significantly in RA patients.

Sequence analysis of a cDNA for D-erythrulose reductase from chicken liver showed that the deduced open reading frame encodes the protein with a molecular mass of 26 kDa consisting of 246 amino acids. Although the reductase shares more than 60% identity in the amino acid sequence with the mouse tetrameric carbonyl reductase, these two enzymes have many biochemical differences; their substrate specificity, subcellular localization, organ distribution, etc. A three-dimensional structure of D-erythrulose reductase was predicted by comparative modeling based on the structure of the tetrameric carbonyl reductase (PDB entry = 1CYD). Most of the residues at the active site (within 4 A from the ligand) of the carbonyl reductase were also conserved in the D-erythrulose reductase. Nevertheless, Val190 and Leu146 in the active site of the tetrameric carbonyl reductase were substituted in the D-erythrulose reductase by Asn192 and His148, respectively. The substitutions in the active sites may be related to the difference in substrate specificity of the two enzymes. The phylogenic analysis of D-erythrulose reductase and the other related proteins suggests that the protein described as a carbonyl reductase D-erythrulose reductase.

Este trabalho refere-se ao controle de uma epidemia de escabiose em aldeia da tribo Panará, localizada no Xingu, Estado do Mato Grosso, Brasil. Descreve as dificuldades encontradas, decorrentes das diferenças culturais entre os investigadores e a população, o processo de conscientização do problema e a participação de toda comunidade no controle da epidemia de escabiose. Relata estratégias de cunho educativo utilizadas e as dificuldades encontradas para conseguir a adesão da comunidade às ações terapêuticas, essenciais ao controle e eliminação da epidemia. Essas estratégias, alicerçadas no ideário da educação em saúde e de respeito aos conhecimentos, valores e necessidades do grupo, previam a participação da comunidade como elemento essencial à consecução dos objetivos dos profissionais de saúde. Elas determinaram o redirecionamento do plano de ação inicial que, por meio de ações conjuntas dos profissionais de saúde, lideranças locais e população alvo, permitiu investigar e controlar a situação inicialmente encontrada. This work reports on an epidemic of scabies in a village of the tribe Panará, located in Xingu, Mato Grosso, Brazil. The educational strategies utilized and the difficulties found to obtain the adhesion of the comunity for the therapeutics actions, essentials to the control of the epidemic are discussed. These strategies, based on an conceptual framework for education in health rooted on the respect for the knowledge, values and necessities of the group, considered the participation of the comunity an essential element for the the adjustment of the initial plan. The actions and combined decisions of health profissionals, local leaderships and target population, made possible the investigation and the control of the epidemic. FUNAI Departamento de Saúde UNIFESP Projeto Xingu da Unidade de Saúde e Meio Ambiente USP Faculdade de Saúde Pública Departamento de Prática de Saúde Pública UNIFESP, Projeto Xingu da Unidade de Saúde e Meio Ambiente SciELO

We have discovered new characteristics of D-erythrulose reductase, namely, that it can catalyze reduction of not only D-erythrulose but also such diketones as diacetyl. These substrates have a common structure with two neighboring carbonyls possibly in s-cis plane structure, showing that the enzyme may rigorously distinguish between substrates and other compounds. D-Erythrulose reductase was predominantly located in the kidney and the liver of the chicken. The obtained results suggest that D-erythrulose reductase plays an important role in metabolizing alpha-dicarbonyls in animal organs, because these diketones widely occur in natural foods.

Doc2 has two C2 domains that interact with Ca2+ and phospholipid. Munc13 has two C2 domains and one C1 domain that interacts with phorbol ester or diacylglycerol (DAG) and phospholipid. Both Doc2 and Munc13 are implicated in Ca2+-dependent neurotransmitter release, but their modes of action still remain unclear. We show here that Doc2 interacts with Munc13 both in a cell-free system and in intact PC12 cells during the high K+-induced Ca2+-dependent exocytosis. The Doc2-Munc13 interactions are stimulated by phorbol ester through the C1 domain of Munc13. Overexpression of the Doc2-interacting domain of Munc13 reduces the Ca2+-dependent exocytosis from PC12 cells, and co-expression with Doc2 suppresses this reduction. These results, together with the earlier findings that secretagogues produce DAG and elevate cytoplasmic Ca2+, suggest that the DAG-induced Doc2-Munc13 interactions play an important role in Ca2+-dependent exocytotic machinery.

Two repeated C 2 -like domains interacting with Ca 2+ and phospholipid are found in synaptotagmin and Rabphilin-3A which are implicated in neurotransmitter release. Here we have isolated a cDNA encoding a novel protein having two repeated C 2 -like domains from a human brain cDNA library. The isolated cDNA encodes a protein with 400 amino acids and a Mr of 44,071. The purified recombinant protein indeed interacts with Ca 2+ and phospholipid. We have named this protein Doc2 ( Do uble C 2 ). Doc2 is exclusively expressed in brain and is highly concentrated in the synaptic vesicle fraction. These results suggest that Doc2 is a novel brain protein and serves as a Ca 2+ sensor in neurotransmitter release.

In addition to the three typical transcripts such as genomic/gag-pol mRNA, env mRNA and tax/rex mRNA, we previously found the singly spliced pX mRNA, termed p21X mRNA, responsible for producing the p21X protein in human T-cell leukemia virus type 1 (HTLV-1)-infected cells. Our finding of the p21X mRNA being constitutively expressed in the fresh peripheral blood mononuclear cells (PBMCs) from patients with ATL has suggested that the expression mechanism is quite different from that of the others. In this paper, the expression mechanism of p21X mRNA was investigated by analyzing the organization of the proviral genomes present in the representative HTLV-1-infected cell lines which are positive or negative for the expression of p21X mRNA. Southern and PCR analyses show that most of the analyzed cell lines contain both one complete and one defective genome each. However, one cell line without the p21X mRNA expression, C91/PL, contains only the complete genome, suggesting that the complete HTLV-1 has no ability to express p21X mRNA in spite of having the ability to produce the infectious virus. The defective genomes of the p21X mRNA positive cell lines, MT-2 and H582, have a large deletion of the entire pol and parts of the gag and env regions including the common domain of the second exon of the doubly spliced tax/rex mRNA, while another defective genome of the p21X mRNA negative cell line, MT-1, has a deletion within the gag-pol gene. We show that these defective genomes have the ability to express their distinct, defective genomic mRNA, suggesting they are active. The defective genomic mRNAs in MT-2 and H582 cells retain the first splice donor and the second splice acceptor sites, suggesting the possibility of synthesizing p21X mRNA by splicing singly with these sites. These findings assume that defective HTLV-1 genomes deleting the second exon region acquire the ability to express p21X mRNA but no ability to express tax/rex mRNA. Such a deletion may explain the difference between the expression mechanisms in the p21X mRNA transcript and those in the other viral transcripts.

Bone resorption in the joints is the characteristic finding in patients with rheumatoid arthritis (RA). Osteoclast-like cells are present in the synovial tissues and invade the bone of patients with RA. The characteristics of these cells are not completely known. In the work reported here, we generated these cells from peripheral-blood monocytes from healthy individuals. The monocytes were co-cultured with nurse-like cells from synovial tissues of patients with RA (RA-NLCs). Within 5 weeks of culture, the monocytes were activated and differentiated into mononuclear cells positive for CD14 and tartrate-resistant acid phosphatase (TRAP). These mononuclear cells then differentiated into multinucleated giant bone-resorbing cells after stimulation with IL-3, IL-5, IL-7, and/or granulocyte-macrophage-colony-stimulating factor. TRAP-positive cells with similar characteristics were found in synovial fluid from patients with RA. These results indicate that multinucleated giant bone-resorbing cells are generated from monocytes in two steps: first, RA-NLCs induce monocytes to differentiate into TRAP-positive mononuclear cells, which are then induced by cytokines to differentiate into multinucleated giant bone-resorbing cells.

Human T-cell leukemia virus type 1 (HTLV-1) remains latent throughout the life of the carrier, with cells containing the provirus and viral gene expression efficiently down-regulated. On a molecular level, exactly how viruses are down-regulated in vivo remains unresolved. We described here the possibility that down-regulation results from the presence of inhibitory elements within the gag-env region of the provirus in fresh peripheral blood mononuclear cells from carriers. In vitro experiments then revealed that potent cis-acting inhibitory elements (CIEs) are indeed contained in two discrete fragments from the pol region and weaker ones in the env region. The effect of CIEs is relieved by the HTLV-1 posttranscriptional regulator Rex through binding to the Rex-responsive element (RxRE), suggesting that Rex might interfere with pre-mRNA degradation and/or activate the export of mRNA molecules harboring both of the inhibitory elements and RxRE on the same RNA molecule. Thus, we propose the hypothesis that such functions of CIEs may be involved in HTLV-1 persistence.

A simulation and experiments have been conducted to develop a two-dimensional position-sensitive detector (PSD) from the previously presented MOS-type PSD. The ac photovoltage phase is used as a positional signal for this MOS-type PSD which requires no reverse-bias voltage, being similar to the Wallmark-type or floating-junction PSD. The simulation has been made using a RC network which equivalently represents the PSD; it has been shown that our PSD is useful for two-dimensional photosensing, admitting a light spot even of nonuniform intensity incident on its face. Experiments have demonstrated that the MOS-type two-dimensional PSD of Al-SiO2-Si has good linearity and electrically adjustable sensitivity. A resolution of 1 µm also has been demonstrated for the present PSD equipped with a simple phase-measuring system.