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The sea cucumber (phylum Echinodermata) body wall is the first line of defense and is well known for its production of secondary metabolites; including vitamins and triterpenoid glycoside saponins that have important ecological functions and potential benefits to human health. The genes involved in the various biosynthetic pathways are unknown. To gain insight into these pathways in an echinoderm, we performed a comparative transcriptome analysis and functional annotation of the body wall and the radial nerve of the sea cucumber Holothuria scabra; to define genes associated with body wall metabolic functioning and secondary metabolite biosynthesis. We show that genes related to signal transduction mechanisms were more highly represented in the H. scabra body wall, including genes encoding enzymes involved in energy production. Eight of the core triterpenoid biosynthesis enzymes were found, however, the identity of the saponin specific biosynthetic pathway enzymes remains unknown. We confirm the body wall release of at least three different triterpenoid saponins using solid phase extraction followed by ultra-high-pressure liquid chromatography-quadrupole time of flight-mass spectrometry. The resource we have established will help to guide future research to explore secondary metabolite biosynthesis in the sea cucumber.

Exposure to solar ultraviolet B (UV-B) is a known causative factor for many skin complications such as wrinkles, black spots, shedding and inflammation. Within the wavelengths 280-320 nm, UV-B can penetrate to the epidermal level. This investigation aimed to test whether extracts from the tropical abalone [Haliotis asinina (H. asinina)] mucus-secreting tissues, the hypobranchial gland (HBG) and gills, were able to attenuate the inflammatory process, using the human keratinocyte HaCaT cell line. Cytotoxicity of abalone tissue extracts was determined using an AlamarBlue viability assay. Results showed that HaCaT cells could survive when incubated in crude HBG and gill extracts at concentrations between <11.8 and <16.9 μg/ml, respectively. Subsequently, cell viability was compared between cultured HaCaT cells exposed to serial doses of UV-B from 1 to 11 (x10) mJ/cm2 and containing 4 different concentrations of abalone extract from both the HBG and gill (0, 0.1, 2.5, 5 μg/ml). A significant increase in cell viability was observed (P<0.001) following treatment with 2.5 and 5 μg/ml extract. Without extract, cell viability was significantly reduced upon exposure to UV-B at 4 mJ/cm2. Three morphological changes were observed in HaCaT cells following UV-B exposure, including i) condensation of cytoplasm; ii) shrunken cells and plasma membrane bubbling; and iii) condensation of chromatin material. A calcein AM-propidium iodide live-dead assay showed that cells could survive cytoplasmic condensation, yet cell death occurred when damage also included membrane bubbling and chromatin changes. Western blot analysis of HaCaT cell COX-2, p38, phospho-p38, SPK/JNK and phospho-SPK/JNK following exposure to >2.5 μg/ml extract showed a significant decrease in intensity for COX-2, phospho-p38 and phospho-SPK/JNK. The present study demonstrated that abalone extracts from the HGB and gill can attenuate inflammatory proteins triggered by UV-B. Hence, the contents of abalone extract, including cellmetabolites and peptides, may provide new agents for skin anti-inflammation, preventing damage due to UV-B.

In decapod crustaceans, the antennal gland (AnG) is a major primary source of externally secreted biomolecules, and some may act as pheromones that play a major role in aquatic animal communication. In aquatic crustaceans, sex pheromones regulate reproductive behaviours, yet they remain largely unidentified besides the N-acetylglucosamine-1,5-lactone (NAGL) that stimulates male to female attraction. In this study, we used an AnG transcriptome of the female giant freshwater prawn (Macrobrachium rosenbergii) to predict the secretion of 226 proteins, including the most abundantly expressed transcripts encoding the Spaetzle protein, a serine protease inhibitor, and an arthropodial cuticle protein AMP 8.1. A quantitative proteome analysis of the female AnG at intermolt, premolt and postmolt, identified numerous proteins of different abundances, such as the hemocyanin subunit 1 that is most abundant at intermolt. We also show that hemocyanin subunit 1 is present within water surrounding females. Of those metabolites identified, we demonstrate that the NAGL and N-acetylglucosamine (NAG) can bind with high affinity to hemocyanin subunit 1. In summary, this study has revealed components of the female giant freshwater prawn AnG that are released and contribute to further research towards understanding crustacean conspecific signalling.

International Perspectives on Sexual and Reproductive Health,2019,45:45-54, https://doi.org/10.1363/45e7819 Metodos: Entre 2013 y 2014, se realizo una encuesta transversal de conocimientos, actitudes y practicas en todo el pals entre 777 vendedores de medicamentos seleccionados al azar; los datos fueron analizados descriptivamente. Se utilize regresion logistica para evaluar las asociaciones entre la exposition a tres intervenciones disenadas para mejorar las practicas de los vendedores de medicamentos (capacitacion conducida por una organization no gubernamental [ONG], un centro de atencion telefonica y capacitacion en el negocio por parte de representantes de las companias farmaceuticas), asl como el conocimiento correcto del regimen de misoprostol solo usado para RM.

International Perspectives on Sexual and Reproductive Health,2019,45:45-54, https://doi.org/10.1363/45e7819 Metodos: Entre 2013 y 2014, se realizo una encuesta transversal de conocimientos, actitudes y practicas en todo el pals entre 777 vendedores de medicamentos seleccionados al azar; los datos fueron analizados descriptivamente. Se utilize regresion logistica para evaluar las asociaciones entre la exposition a tres intervenciones disenadas para mejorar las practicas de los vendedores de medicamentos (capacitacion conducida por una organization no gubernamental [ONG], un centro de atencion telefonica y capacitacion en el negocio por parte de representantes de las companias farmaceuticas), asl como el conocimiento correcto del regimen de misoprostol solo usado para RM.

CONTEXT: In Bangladesh, prior to the availability of the approved combination regimen of mifepristone and misoprostol for menstrual regulation (MR), drug seller provision of misoprostol-only regimens for MR without a prescription was widespread but service quality was poor. Examining provider practices relating to misoprostol-only provision in Bangladesh may increase understanding of misoprostol use and provision in other low-resource, legally restrictive settings. METHODS: In 2013–2014, a countrywide cross-sectional knowledge, attitudes and practice survey was conducted among 777 randomly selected drug sellers; data were analyzed descriptively. Logistic regression was used to test the associations between exposure to three interventions designed to improve drug seller practice (nongovernmental organization [NGO]–led training, a call center and in-shop training from pharmaceutical company representatives) and correct knowledge of the misoprostol-only MR regimen. RESULTS: Almost all (97%) of the drug sellers reported providing medications intended for MR; misoprostol-only was more commonly sold than the combination regimen (96% vs. 26%). Nine percent had received NGO–led training, 62% had received in-shop training from a pharmaceutical company representative and 27% had used the call center. Overall, 19% of drug sellers knew the correct misoprostol-only MR regimen, and 74% wanted more information about this regimen. Correct regimen knowledge was positively associated with receipt of NGO training and call center utilization (odds ratios, 2.0 and 1.9, respectively). CONCLUSIONS: NGO–led training and call centers should be considered in other settings in which misoprostol alone is provided off-label for pregnancy termination.

The purpose of the study was to determine the concentration of serum trace and other essential elements of generalized anxiety disorder patients and to find out the relationship between element levels and nutritional status or socioeconomic factors. The study was conducted among 50 generalized anxiety disorder patients and 51 healthy volunteers. Patients were selected and recruited in the study with the help of a clinical psychologist by random sampling. The concentrations of serum trace elements (Zn, Cu, Mn, and Fe) and other two essential elements (Ca and Mg) were determined by graphite furnace and flame atomic absorption spectroscopy. Data were analyzed by independent t test, Pearson’s correlation analysis, regression analysis, and analysis of variance. The serum concentrations of Zn, Cu, Mn, Fe, Ca, and Mg in generalized anxiety disorder patients were 1.069 ± 0.40, 1.738 ± 0.544, 1.374 ± 0.750, 3.203 ± 2.065, 108.65 ± 54.455, and 21 ± 4.055 mg/L, while those were 1.292 ± 0.621, 0.972 ± 0.427, 0.704 ± 0.527, 1.605 ± 1.1855, 101.849 ± 17.713, and 21.521 ± 3.659 mg/L in control subjects. Significantly decreased (p < 0.05) serum Zn concentration was found in the patient group compared to the control group while serum level of Cu, Mn, and Fe was significantly (p < 0.05) higher, but the differences of the concentration of Ca and Mg between the patient and control groups were not significant (p > 0.05). Socioeconomic data revealed that most of the patients were in the lower middle class group and middle-aged. Mean BMI of the control group (23.63 ± 3.91 kg/m²) and the patient group (23.62 ± 3.77 kg/m²) was within the normal range (18.5–25.0 kg/m²). The data obtained from different interelement relations in the generalized anxiety disorder patients and control group strongly suggest that there is a disturbance in the element homeostasis. So changes in the serum trace element level in generalized anxiety disorder patients occur independently and they may provide a prognostic tool for the diagnosis and treatment of this disease.

The cell wall structure of gram-negative bacteria is more complex than that of gram-positive bacteria because of the presence of two layers external to the cytoplasmic membrane. Purpose of this study is to compare the four different methods for genomic DNA extraction from three gram negative strains-Escherichia coli, Klebsiella species, and Salmonella typhi. The genomic DNA was extracted from gram-negative bacteria cultures using enzymatic lysis with Proteinase-K, chemical lysis with cetyl trimethyl ammonium bromide (CTAB), physical lysis with Triton X-100 and Beads beating methods. The yield and quality of DNA were compared by spectrophotometry and agarose gel electrophoresis. The genomic DNA purity ratio was found within 1.1-1.9 for all four methods. This study revealed that, the chemical lysis with CTAB was the most efficient method for extracting genomic DNA from gram negative strains. In terms of chemical lysis with CTAB for Salmonella typhi and Klebsiella species the highest purity ratio were 1.9 and 1.8, respectively.