Untargeted-based metabolomics analysis and in vitro/in silico antiviral activity of extracts from Phyllanthus brasiliensis (Aubl.) Poir.

INTRODUCTION: This study describes the molecular profile and the potential antiviral activity of extracts from Phyllanthus brasiliensis, a plant widely found in the Brazilian Amazon. The research aims to shed light on the potential use of this species as a natural antiviral agent. METHODS: The extracts were analysed using liquid chromatography-mass spectrometry (LC-MS) system, a potent analytical technique to discover drug candidates. In the meantime, in vitro antiviral assays were performed against Mayaro, Oropouche, Chikungunya, and Zika viruses. In addition, the antiviral activity of annotated compounds was predicted by in silico methods. RESULTS: Overall, 44 compounds were annotated in this study. The results revealed that P. brasiliensis has a high content of fatty acids, flavones, flavan-3-ols, and lignans. Furthermore, in vitro assays revealed potent antiviral activity against different arboviruses, especially lignan-rich extracts against Zika virus (ZIKV), as follows: methanolic extract from bark (MEB) [effective concentration for 50% of the cells (EC50 ) = 0.80 µg/mL, selectivity index (SI) = 377.59], methanolic extract from the leaf (MEL) (EC50 = 0.84 µg/mL, SI = 297.62), and hydroalcoholic extract from the leaf (HEL) (EC50 = 1.36 µg/mL, SI = 735.29). These results were supported by interesting in silico prediction, where tuberculatin (a lignan) showed a high antiviral activity score. CONCLUSIONS: Phyllanthus brasiliensis extracts contain metabolites that could be a new kick-off point for the discovery of candidates for antiviral drug development, with lignans becoming a promising trend for further virology research.

The association between 11C-methionine uptake, IDH gene mutation, and MGMT promoter methylation in patients with grade II and III gliomas.

AIM: To evaluate the association between 11C-methionine positron-emission tomography (11C-methionine PET) findings, isocitrate dehydrogenase (IDH) gene mutation, and O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation in patients with grade II and III gliomas. MATERIALS AND METHODS: Data were collected from 40 patients with grade II and III gliomas who underwent both magnetic resonance imaging (MRI) and 11C-methionine PET as part of their pre-surgical examination. IDH mutation was examined via DNA sequencing, and MGMT promoter methylation via quantitative methylation-specific polymerase chain reaction (PCR). RESULTS: A threshold of MGMT promoter methylation of 1% was significantly associated with tumour/normal tissue (T/N) ratio. The T/N ratio in samples with MGMT promoter methylation ≥1% was higher than that in samples with MGMT promoter methylation <1%, and the difference was statistically significant (p=0.011). Reliable prediction of MGMT promoter methylation (<1% versus ≥1%) was possible using the T/N ratio under the receiver operator characteristic (ROC) curve with a sensitivity and specificity of 75% each (cut-off value=1.6: p=0.0226, area under the ROC curve [AUC]=0.76172). Conversely, the T/N ratio had no association with IDH mutation (p=0.6). The ROC curve revealed no reliable prediction of IDH mutation using the T/N ratio (p=0.606, AUC=0.60577). CONCLUSION: 11C-methionine PET parameters can predict MGMT promoter methylation but not IDH mutation status. 11C-methionine uptake may have limited potential to reflect DNA methylation processes in grade II and III gliomas.

Secure and Sustainable Sourcing of Plant Tissues for the Exhaustive Exploration of Their Chemodiversity.

The main challenge of plant chemical diversity exploration is how to develop tools to study exhaustively plant tissues. Their sustainable sourcing is a limitation as bioguided strategies and dereplication need quite large amounts of plant material. We examine if alternative solutions could overcome these difficulties by obtaining a secure, sustainable, and scalable source of tissues able to biosynthesize an array of metabolites. As this approach would be as independent of the botanical origin as possible, we chose eight plant species from different families. We applied a four steps culture establishment procedure, monitoring targeted compounds through mass spectrometry-based analytical methods. We also characterized the capacities of leaf explants in culture to produce diverse secondary metabolites. In vitro cultures were successfully established for six species with leaf explants still producing a diversity of compounds after the culture establishment procedure. Furthermore, explants from leaves of axenic plantlets were also analyzed. The detection of marker compounds was confirmed after six days in culture for all tested species. Our results show that the first stage of this approach aiming at easing exploration of plant chemodiversity was completed, and leaf tissues could offer an interesting alternative providing a constant source of natural compounds.

Histopathological study of carcinoma showing thymus-like differentiation (CASTLE).

INTRODUCTION: Carcinoma showing thymus-like differentiation (CASTLE) is a rare tumour that mainly arises from the thyroid gland, or occasionally, from the head and neck. Although the 10-year survival rate of patients with CASTLE is approximately 80%, local recurrence and distant metastasis are observed in some cases. A recent systematic review for CASTLE indicated that the prognostic factors are treatment-dependent, and postoperative radiotherapy significantly improves patient survival. CASE REPORT: Herein, we describe and compare three cases of CASTLE, including a case with distant metastasis despite administering postoperative chemotherapy. Thus, the mechanisms underlying metastasis of CASTLE are unclear. This case study helps to elucidate the histopathological risk factors of metastasis in CASTLE. DISCUSSION: We found that prominent lymphovascular invasion and higher proliferative activities might be risk factors of metastasis in CASTLE. In addition, we have summarised the cytological, morphological, and immunohistochemical features of CASTLE for an accurate diagnosis.

Primary solitary fibrous tumour of the prostate: A case report and literature review.

INTRODUCTION: Solitary fibrous tumour (SFT) is a rare mesenchymal tumour with intermediate malignant potential. Although this tumour arises in several sites, prostatic SFT is an extremely rare neoplasm and may prove confusing owing to the lack of clinical experience because of tumour rarity. The diagnosis may be further difficult because SFTs can manifest positive immunoreactivity for CD34 and progesterone receptor, which are known markers of prostatic stromal tumours. Herein, we describe a case of prostatic SFT that was difficult to differentiate from a prostatic stromal tumour of uncertain malignant potential because of positive immunoreactivity to CD34 and progesterone receptor. CASE REPORT: A 40-year-old Japanese man presented with lower abdominal pain. Computed tomography revealed a prostatic mass; furthermore, prostate core needle biopsy revealed proliferating bland spindle cells, without necrosis or prominent mitoses. Tumour cells were positive for CD34 and progesterone receptor on immunohistochemical analysis; thus, a prostatic stromal tumour of uncertain malignant potential was initially suspected. However, as the tumour cells showed positive immunoreactivity for STAT6, the final diagnosis was an SFT of the prostate. The patient underwent tumour resection, and at the 6-month postoperative follow-up, neither local recurrence nor distant metastasis occurred. CONCLUSION: For an accurate diagnosis of an SFT of the prostate, STAT6 immunohistochemistry should be conducted for all mesenchymal tumours of the prostate. When STAT6 immunohistochemical analysis is unfeasible, pathologists should be aware that the morphological and immunohistochemical characteristics of SFT variable from case to case and diagnose with combined analysis of several immunohistochemical markers.

Hematological and anthelminthic responses of tambaqui (Colossoma macropomum) supplemented with Artrhospira platensis and Chlorella vulgaris.

The present study evaluated the hematological, antiparasitic and growth responses in tambaqui (Colossoma macropomum) fed with diets supplemented with the microalgae Arthrospira platensis and Chlorella vulgaris (0%; 10% A. platensis; 10% C. vulgaris; and 5% A. platensis+5% C. vulgaris). Tambaqui (n=60, 62.57 ± 8.76 g) were fed for 20 days with experimental diets. Blood samples collection was done to determine hematological parameters, and gills were removed to identify and count monogenetic parasites. Supplementation with A. platensis 10% reduced red blood cells count, in consequence mean corpuscular volume and mean hemoglobin concentration increased. Total leukocyte, monocyte, eosinophil, and basophil counts reduced with the use of A. platensis. Higher monocytes, eosinophil, and basophil numbers in tambaqui fed with diet supplemented with 10% C. vulgaris were observed and may have been due to the presence of immunostimulants in this microalga composition. Reduction on total cholesterol in tambaqui that received both microalgae (A. platensis 5%+C. vulgaris 5%) may indicate that combined supplementation presented greater benefits to the health for C. macropomum than separately. Both microalgae were efficient against monogenetic parasites of tambaqui. Thus, the dietary use of the microalgae A. platensis and C. vulgaris provided immunostimulant and antiparasitic efficacy in C. macropomum.

Exploring the antiviral potential of justicidin B and four glycosylated lignans from Phyllanthus brasiliensis against Zika virus: A promising pharmacological approach.

BACKGROUND: Zika virus (ZIKV) is an emerging arbovirus that in recent years has been associated with cases of severe neurological disorders, such as microcephaly in newborns and Guillain-Barré syndrome in adults. As there is no vaccine or treatment, the search for new therapeutic targets is of great relevance. In this sense, plants are extremely rich sources for the discovery of new bioactive compounds and the species Phyllanthus brasiliensis (native to the Amazon region) remains unexplored. PURPOSE: To investigate the potential antiviral activity of compounds isolated from P. brasiliensis leaves against ZIKV infection. METHODS: In vitro antiviral assays were performed with justicidin B (a lignan) and four glycosylated lignans (tuberculatin, phyllanthostatin A, 5-O-ß-d-glucopyranosyljusticidin B, and cleistanthin B) against ZIKV in Vero cells. MTT colorimetric assay was used to assess cell viability and plaque forming unit assay to quantify viral load. In addition, for justicidin B, tests were performed to investigate the mechanism of action (virucidal, adsorption, internalization, post-infection). RESULTS: The isolated compounds showed potent anti-ZIKV activities and high selectivity indexes. Moreover, justicidin B, tuberculatin, and phyllanthostatin A completely reduced the viral load in at least one of the concentrations evaluated. Among them, justicidin B stood out as the main active, and further investigation revealed that justicidin B exerts its antiviral effect during post-infection stages, resulting in a remarkable 99.9 % reduction in viral load when treatment was initiated 24 h after infection. CONCLUSION: Our findings suggest that justicidin B inhibits endosomal internalization and acidification, effectively interrupting the viral multiplication cycle. Therefore, the findings shed light on the promising potential of isolated compounds isolated from P. brasiliensis, especially justicidin B, which could contribute to the drug development and treatments for Zika virus infections.

Margaritaria nobilis L.f. (Phyllanthaceae) Ethanolic Extract: Low Acute Oral Toxicity and Antinociceptive Activity.

Margaritaria nobilis L.f. (Phyllanthaceae), a native Brazilian tree occurring mainly in the Amazon, is used in folk medicine for the treatment of abscesses (bark) and cancer-like symptoms (leaves). The present study evaluates the safety of its acute oral administration and its effects on nociception and plasma leakage. The chemical constitution of the leaf's ethanolic extract is determined by ultra-performance liquid chromatography-high-resolution mass spectrometry (LC-MS. Its acute oral toxicity is evaluated in female rats at a dose of 2000 mg/kg, evaluating the occurrence of deaths and Hippocratic, behavioral, hematological, biochemical, and histopathological changes, as well as food and water consumption and weight gain. Antinociceptive activity is evaluated in male mice with acetic-acid-induced peritonitis (APT) and formalin (FT) tests. An open field (OF) test is performed to verify possible interferences in the animals' consciousness or locomotion. LC-MS analysis shows the presence of 44 compounds classified as phenolic acid derivatives, flavonoids and O-glycosylated derivatives, and hydrolyzable tannins. No deaths or significant behavioral, histological, or biochemical changes are observed in the toxicity assessment. In nociception tests, M. nobilis extract significantly reduces abdominal contortions in APT, demonstrating selectivity for inflammatory components (FT second phase), not interfering in neuropathic components (FT first phase) or consciousness and locomotion levels in OF. Additionally, M. nobilis extract inhibits plasma acetic-acid-induced leakage. These data demonstrate the low toxicity of M. nobilis ethanolic extract, as well as its effectiveness in modulating inflammatory nociception and plasma leakage, possibly related to the flavonoids and tannins present in its composition.

Physiological responses of tambaqui (Colossoma macropomum) fed diets supplemented with silage from fish and vegetables residues.

The demand for products to replace high-cost raw materials, such oil and fish meal, in the manufacture of feed for use in aquaculture, while also guaranteeing the nutritional quality of the diets, is increasing. Silage produced with fish and vegetables residues is a low-cost and efficient protein source. The objective of the present study was to evaluate the physiological and biochemical responses of tambaqui fingerlings fed four different levels of silage included in commercial feed with 28% crude protein, over two periods: 45 and 90 days. Each treatment was carried out over three replications, with 10 tambaqui in each 100 L experimental tank. At the end of each established period, blood samples were collected from five animals from each repetition to determine the hematological and biochemical variables. Body weight and total length, hepatosomatic and liposomal indices and hematocrit of specimens fed with diets supplemented with silage did not exhibit significant changes in both assessment period. After 45 days of feeding, the hemoglobin concentration increased when tambaqui were fed a diet including 20% silage. The red blood cell count, mean corpuscular volume and mean corpuscular hemoglobin did not change between treatments in either period. The total protein concentrations increased significantly in the plasma of tambaqui fed with diets with the inclusion of 5 and 10% of silage, evaluated after feeding for 45 days. It was found that the groups which had silage included in their diet did not exhibit significant alterations in the evaluated parameters, and the diet was therefore not consider harmful to the health of tambaqui. Therefore, the use of silage as a feed supplement during tambaqui farming is a sustainable alternative for producers, as it leads to a reduction of impacts of fish and vegetables waste disposal.

Measurement of adiponectin production from differentiated metabolic stem cells.

To treat metabolic syndrome, fat tissue dysfunction should be corrected rather than controlling conventional risk factors such as hypertension, dyslipidemia, and diabetes mellitus. For this purpose, accumulating evidence suggests increasing plasma adiponectin levels can be a key treatment strategy, especially in setting of food or drug selection. Here we report that adipocyte precursors obtained from several sites of fat tissue, which we call Metabolic Stem Cells (MSC), could be used as a novel screening system to identify adiponectin enhancing drugs or food for individual patients. MSC were prepared from fat tissues collected from 29 patients. They were differentiated in cultures into mature adipocytes. The time course of adiponectin production was independent of the number of mature adipocytes and gradually decreased at 48 h after differentiation. Pioglitazone, a full PPARgamma agonist, stabilized adiponectin production at days 8-16 after differentiation, whereas telmisartan, a partial PPARgamma agonist, showed variable response. Dividing the adiponectin secretion of day 12 by that of day 10 provided an estimate of adiponectin-producing activity irrespective of the number of MSC-derived adipocytes in culture. Using this score of adiponectin-production activity, we successfully assessed 16 agents in a 96-well plate. The effect of each agent on adiponectin production showed a similar pattern, independent of the site of isolated adipose tissue. Our results show that MSC can be used as a tool for selecting drugs that enhance adiponectin-production activity.

Time trends in suicide rates by domestic gas or car exhaust gas inhalation in Japan, 1968-1994.

AIMS: A reduction in the carbon monoxide content of domestic gas and car exhaust gas has been associated with a decrease in gassing suicides in many western countries. In Japan, a reduction in the carbon monoxide content of domestic gas supply began in the early 1970s, and carbon monoxide emissions standards of new passenger cars were significantly strengthened in 1978. However, little is known about the impact of detoxification of these gases on gassing-related suicides in Japan. Therefore, we examined the changing patterns of suicide due to domestic gas or car exhaust gas inhalation by gender and age in Japan between 1968 and 1994. METHODS: Suicide mortality data were obtained from the Vital Statistics of Japan. In this study, age was divided into four groups: 15-24, 25-44, 45-64 and 65+ years. Method of suicide was divided into three groups: domestic gas, car exhaust gas and non-gases. We calculated method-specific age-standardised suicide rates by gender within each of the four age groups. We applied joinpoint regression to the data and quantified the observed changes. RESULTS: Suicide rates by domestic gas, regardless of gender and age, increased from 1968 to the mid-1970s and then decreased sharply. The proportion of all suicides accounted for by domestic gas was comparatively high in the mid-1970s among females aged 15-24 and 25-44 years, while for other gender-age-groups the proportion of domestic gas suicides remained small, even at the peak. For females aged 15-44 years, the decrease in domestic gas suicides appeared to cause a substantial decrease in overall suicides in this gender/age group. Car exhaust gas was a more common method for males, particularly those aged 25-64 years. Car exhaust gas suicide rates for males aged 25-64 years peaked in the mid-1980s, followed by a sharp decrease. CONCLUSIONS: A reduction in the carbon monoxide content of the domestic gas, which began in the early 1970s in Japan, was associated with a decrease in domestic gas suicides for both genders of all ages. Concerning females aged 15-44 years, a decrease in domestic gas suicides caused a substantial decrease in overall suicides in this gender/age group since the proportion of domestic gas suicides among all suicides combined was comparatively large. However, it remains uncertain whether the introduction of catalytic converters in the 1970s in Japan resulted in a reduction of suicides from car exhaust gas inhalation.

Physiological responses of tambaqui (Colossoma macropomum) fed diets supplemented with silage from fish and vegetables residues / Respostas fisiológicas de tambaqui (Colossoma macropomum) alimentado com dietas suplementadas com silagem de resíduos de pescado e vegetais

The demand for products to replace high-cost raw materials, such oil and fish meal, in the manufacture of feed for use in aquaculture, while also guaranteeing the nutritional quality of the diets, is increasing. Silage produced with fish and vegetables residues is a low-cost and efficient protein source. The objective of the present study was to evaluate the physiological and biochemical responses of tambaqui fingerlings fed four different levels of silage included in commercial feed with 28% crude protein, over two periods: 45 and 90 days. Each treatment was carried out over three replications, with 10 tambaqui in each 100 L experimental tank. At the end of each established period, blood samples were collected from five animals from each repetition to determine the hematological and biochemical variables. Body weight and total length, hepatosomatic and liposomal indices and hematocrit of specimens fed with diets supplemented with silage did not exhibit significant changes in both assessment period. After 45 days of feeding, the hemoglobin concentration increased when tambaqui were fed a diet including 20% silage. The red blood cell count, mean corpuscular volume and mean corpuscular hemoglobin did not change between treatments in either period. The total protein concentrations increased significantly in the plasma of tambaqui fed with diets with the inclusion of 5 and 10% of silage, evaluated after feeding for 45 days. It was found that the groups which had silage included in their diet did not exhibit significant alterations in the evaluated parameters, and the diet was therefore not consider harmful to the health of tambaqui. Therefore, the use of silage as a feed supplement during tambaqui farming is a sustainable alternative for producers, as it leads to a reduction of impacts of fish and vegetables waste disposal.

A procura por insumos que substituam produtos de alto custo, como óleo e farinha de peixe, na fabricação de rações para uso na aquicultura é crescente, sendo necessário garantir a qualidade nutricional das dietas. A silagem produzida a partir de resíduos de pescado e de vegetais apresenta-se como uma alternativa de baixo custo e eficiente fonte proteica. O objetivo do presente estudo foi avaliar as respostas fisiológicas e bioquímicas de alevinos de tambaqui alimentados com quatro níveis de inclusão de silagem em ração comercial com 28% de proteína bruta, em dois períodos: 45 e 90 dias. Cada tratamento foi realizado em três repetições, com 10 tambaquis em cada caixa experimental de 100 L. Ao término de cada período estabelecido, amostras sanguíneas foram coletadas de cinco animais de cada repetição para determinação das variáveis hematológicas e bioquímicas. Peso, comprimento total e índices hepatossomático e lipossomático de espécimes alimentados com silagem não mostraram alterações significativas em ambos os períodos de avaliação, bem como os valores de hematócrito. A concentração de hemoglobina de tambaqui após 45 dias de alimentação aumentou quando foi fornecida dieta com inclusão de 20% de silagem. A contagem de eritrócitos, volume corpuscular médio e hemoglobina corpuscular média não apresentaram alterações entre os tratamentos, em ambos os períodos. As proteínas totais aumentaram significativamente no plasma de tambaquis que receberam dietas com inclusão de 5 e 10% de silagem, avaliados após 45 dias de alimentação. Evidenciou-se que os grupos com inclusão de silagem na dieta não apresentaram alterações significativas nos parâmetros avaliados, assim não sendo prejudiciais à higidez do tambaqui. Portanto, o uso da silagem como suplemento alimentar durante o cultivo do tambaqui é uma alternativa sustentável para produtores, por promover a redução do descarte de resíduos de pescado e da agricultura.

Physiological responses of tambaqui (Colossoma macropomum) fed diets supplemented with silage from fish and vegetables residues

Abstract The demand for products to replace high-cost raw materials, such oil and fish meal, in the manufacture of feed for use in aquaculture, while also guaranteeing the nutritional quality of the diets, is increasing. Silage produced with fish and vegetables residues is a low-cost and efficient protein source. The objective of the present study was to evaluate the physiological and biochemical responses of tambaqui fingerlings fed four different levels of silage included in commercial feed with 28% crude protein, over two periods: 45 and 90 days. Each treatment was carried out over three replications, with 10 tambaqui in each 100 L experimental tank. At the end of each established period, blood samples were collected from five animals from each repetition to determine the hematological and biochemical variables. Body weight and total length, hepatosomatic and liposomal indices and hematocrit of specimens fed with diets supplemented with silage did not exhibit significant changes in both assessment period. After 45 days of feeding, the hemoglobin concentration increased when tambaqui were fed a diet including 20% silage. The red blood cell count, mean corpuscular volume and mean corpuscular hemoglobin did not change between treatments in either period. The total protein concentrations increased significantly in the plasma of tambaqui fed with diets with the inclusion of 5 and 10% of silage, evaluated after feeding for 45 days. It was found that the groups which had silage included in their diet did not exhibit significant alterations in the evaluated parameters, and the diet was therefore not consider harmful to the health of tambaqui. Therefore, the use of silage as a feed supplement during tambaqui farming is a sustainable alternative for producers, as it leads to a reduction of impacts of fish and vegetables waste disposal.

Resumo A procura por insumos que substituam produtos de alto custo, como óleo e farinha de peixe, na fabricação de rações para uso na aquicultura é crescente, sendo necessário garantir a qualidade nutricional das dietas. A silagem produzida a partir de resíduos de pescado e de vegetais apresenta-se como uma alternativa de baixo custo e eficiente fonte proteica. O objetivo do presente estudo foi avaliar as respostas fisiológicas e bioquímicas de alevinos de tambaqui alimentados com quatro níveis de inclusão de silagem em ração comercial com 28% de proteína bruta, em dois períodos: 45 e 90 dias. Cada tratamento foi realizado em três repetições, com 10 tambaquis em cada caixa experimental de 100 L. Ao término de cada período estabelecido, amostras sanguíneas foram coletadas de cinco animais de cada repetição para determinação das variáveis hematológicas e bioquímicas. Peso, comprimento total e índices hepatossomático e lipossomático de espécimes alimentados com silagem não mostraram alterações significativas em ambos os períodos de avaliação, bem como os valores de hematócrito. A concentração de hemoglobina de tambaqui após 45 dias de alimentação aumentou quando foi fornecida dieta com inclusão de 20% de silagem. A contagem de eritrócitos, volume corpuscular médio e hemoglobina corpuscular média não apresentaram alterações entre os tratamentos, em ambos os períodos. As proteínas totais aumentaram significativamente no plasma de tambaquis que receberam dietas com inclusão de 5 e 10% de silagem, avaliados após 45 dias de alimentação. Evidenciou-se que os grupos com inclusão de silagem na dieta não apresentaram alterações significativas nos parâmetros avaliados, assim não sendo prejudiciais à higidez do tambaqui. Portanto, o uso da silagem como suplemento alimentar durante o cultivo do tambaqui é uma alternativa sustentável para produtores, por promover a redução do descarte de resíduos de pescado e da agricultura.

Time trends in method-specific suicide rates in Japan, 1990-2011.

BACKGROUND: Little is known about whether particular suicide methods have contributed differently to the recent unfavourable suicide mortality trends in Japan. Analysing such trends may shed light on the effect of potentially preventable factors, such as the impact of restricting access to certain popular suicide methods, on overall rates. Therefore, we assessed recent trends in method-specific suicide by gender and age in Japan. METHOD: Suicide mortality and population data between 1990 and 2011 were obtained from the Vital Statistics of Japan and used to calculate method-specific mortality rates. Suicide methods were divided into seven groups: overdose, gases, hanging, drowning, cutting, jumping and other means. Age was divided into four groups: 15-24, 25-44, 45-64 and 65+ years. We applied joinpoint regression to the data and quantified the observed changes. RESULTS: The results of the joinpoint regression analyses showed a sharp increase in overall suicide rates for males and females of all ages until the late 1990s. Suicide from hanging and jumping, in particular, contributed to this increase. After 2000, an increasing trend in overall suicide rates in both males and females aged 15-24 and 25-44 years was observed, with overdose, gases and hanging contributing to this increasing trend. CONCLUSIONS: Our findings revealed that different suicide methods varied in their contribution to the recent overall suicide transition in Japan. Regarding factors associated with the recent increase in suicides by overdose, gases, hanging and jumping, further research is needed in order to promote and implement effective means restriction strategies.

Relationships of C-reactive protein, uric acid, and glomerular filtration rate to arterial stiffness in Japanese subjects.

The relationships between C-reactive protein (CRP), uric acid (UA), glomerular filtration rate (GFR), and arterial stiffness have not been fully investigated. The aim of this study was to clarify whether CRP, UA, and estimated GFR are related to arterial stiffness estimated using brachial-ankle pulse wave velocity (baPWV). The subjects were local government employees (3412 men and 854 women). baPWV, CRP, UA, GFR, and conventional risk factors were evaluated. Multiple linear regression analyses revealed that CRP and UA were significantly related to an elevation of PWV in male and female subjects, and that the estimated GFR was significantly related to an elevation of PWV in male subjects. Significant progressive increases in baPWV were observed across the quartiles of CRP in male subjects and for UA in male and female subjects. In female subjects, the relationship of quartile CRP to baPWV had marginal significance (P = 0.055). But, in male and female subjects, quartile of estimated GFR had no significant association with PWV. These results suggest that CRP and UA are associated with an increase of arterial stiffness in male and female subjects, and that estimated GFR is possibly related to arterial stiffness in male subjects.

Data on the sites of stimulatory feedback action of gonadal steroids indispensable for luteinizing hormone release in the rat.

The sites of the stimulatory feedback action of gonadal steroids on LH release were investigated in estrogen-primed ovariectomized rats. Either estradiol benzoate (E2) or estrone (E1) injections 72 h after E2 priming induced a significant increase in serum LH 30 h later, whereas injections of progesterone (P) did so 6 h later. Horizontal sections placed above the medial preoptic area prevented the increase after E2 injections but not after E1 and P injections. Retrochiasmatic sections or bilateral lesions placed in the medial-basal part of the suprachiasmatic area prevented any increases in serum LH induced by steroid injections. Intracerebral implantations of E2 and P in E2-primed ovariectomized rats induced a similar significant increase in serum LH when E2 was implanted into the bed nucleus of stria terminalis (BST), the lateral septum (l-SEPT) and the preoptic suprachiasmatic area (POSC), and when P was implanted in to the medial amygdala, the diagonal band of Broca (DBB), the POSC, the l-SEPT and the anterior hypothalamic area (AHA). These results suggest that the stimulatory feedback effect of E2 is exerted on the limbic structures, especially the BST and the l-SEPT, while the main sites of the stimulatory feedback action of P are located in the DBB, the POSC, and the AHA.

Data on the sites of the stimulatory feedback action of gonadal steroids indispensable for follicle-stimulating hormone release in the rat.

The sites of the stimulatory feedback action of gonadal steroids on FSH release were examined in estrogen-primed ovariectomized rats. Estradiol benzoate (E2) injected 72 h after E2 priming induced a significant increase in serum FSH 6 and 30 h later. However, injections of estrone failed to induce a similar significant increase. Progesterone (P) injections also increased serum FSH 6 h later. Horizontal sections placed above the medial preoptic area prevented the increase in serum FSH after E2 injections but not after P injections. Furthermore, retrochiasmatic sections or bilateral lesions in the medial basal part of the suprachiasmatic area prevented the increase after E2 injections and P injections. E2, implanted into the bed nucleus of stria terminalis and the anterior hypothalamic area in E2-primed ovariectomized rats, induced a significant increase and decrease in serum FSH, respectively, whereas P implanted into the same areas failed to induce it. On the other hand, P implantations into the diagonal band of Broca and the lateral and the medial septum were effective in increasing and decreasing serum FSH, respectively, whereas E2 implantations into these areas were ineffective. Neither E2 nor P implanted into the medial amygdala, the medial preoptic area, or the hippocampus altered serum FSH levels. These results suggest that the stimulatory feedback actions of E2 and P on FSH release are exerted at different sites in the limbic structures.