Studi Karakteristik Tailing Pada Lokasi Eks Penambangan Timah di Bukit Sambung Giri Kecamatan Merawang Kabupaten Bangka

Tailing deposits at ex-tin mining area in Bangka and Belitung Island has a large number of economical residual minerals. This study is to identify tailing characteristic and its potential mineralogy and elements based on shallow hand boring (Auger) samples, along stream line in west part of Sambung Giri Hill. Every coordinate sample plotted on base map. Loboratory analisys using Strereo Mikroscope for identify minerals characteristics, and geochemical analysis using XRF for identification the presence of elements. The tailing deposits has laterally grain size gradation to down stream, composed above 94% sand fraction, and small amount of shale and granule fraction. Quartz mineral presence more then 90%, followed by iron oxide (hematite), clay, zircon, cassiterite, ilmenite, monazite and tourmaline minerals, showed low consistency in lateral spatial exept quartz, with its Rare Earth Elementh (REE) composition is Cerium (Ce), Lanthanum (La), Yttrium (Y), Neodymium (Nd) and Samarium (Sm).

β-adrenoceptor activation increased VAMP-2 and syntaxin-4 in secretory granules are involved in protein secretion of submandibular gland through the PKA/F-actin pathway

Autologous submandibular gland transplantation is an effective treatment for severe dry eye syndrome. However, the protein secretion in transplanted gland is altered by a mechanism that remains to be elucidated. In the present study, we found that β1-adrenoceptor (β1-AR) and β2-AR expression and the phosphorylation of the downstream molecule protein kinase A (PKA) were elevated in transplanted submandibular glands obtained from epiphora patients. Synaptobrevin/vesicle-associated membrane protein 2 (VAMP-2) interacted with syntaxin-4 and actin in human submandibular gland. The contents of syntaxin-4 and actin interacting with VAMP-2 were increased in transplanted gland. Moreover, VAMP-2 and syntaxin-4 expression in the secretory granule fraction, and VAMP-2 expression in the membrane protein fraction were increased in isoproterenol-treated and transplanted glands. Isoproterenol increased F-actin polymerization in the apical and lateral regions of the cytoplasm in both control and transplanted glands. Inhibiting PKA activity and/or F-actin formation abolished the isoproterenol-enhanced expression of VAMP-2 and syntaxin-4 in the secretory granule fraction and the isoproterenol-enhanced expression of VAMP-2 in the membrane protein fraction. Taken together, these results indicate that the activation of β-ARs induces secretory granules and cell membrane fusion via the interaction of VAMP-2 and syntaxin-4 in a PKA- and F-actin-dependent manner in human submandibular gland. Up-regulated β-ARs might participate in altering protein secretion in transplanted submandibular gland by promoting the interaction of VAMP-2 with syntaxin-4.

Promotion of partial nitritation-anammox process by improving granule proportion

For enhancing the partial nitritation-anammox (PN/A) process, the effects of granule fraction on system performance were investigated in this study. Two sequencing batch reactors (SBRs) were inoculated with PN/A biomass with a floc mass fraction of 53%. In SBR1, when the nitrogen removal rate (NRR) was stable, flocculent sludge was gradually discharged from the reactor using a screen, and the granule fraction was therefore increased. However, nitrogen removal was not improved and finally deteriorated due to the loss of nitritation activity. In SBR2, most flocculent sludge was eliminated and granular proportion was maintained at over 90% by controlling a short settling and decanting time. NRR was low initially but gradually improved to 1.23 kg N/(m3·d), which was 54% higher than SBR1. Ammonium oxidation activities of flocs and granules were respectively measured. Results suggested that the increase of nitritation activity in the granules was the main reason for the improvement of nitrogen removal in SBR2.

Calcium- and polyphosphate-containing acidic granules of sea urchin eggs are similar to acidocalcisomes, but are not the targets for NAADP

Acidocalcisomes are acidic calcium-storage compartments described from bacteria to humans and characterized by their high content in poly P (polyphosphate), a linear polymer of many tens to hundreds of Pi residues linked by high-energy phosphoanhydride bonds. In the present paper we report that millimolar levels of short-chain poly P (in terms of Pi residues) and inorganic PPi are present in sea urchin extracts as detected using 31P-NMR, enzymatic determinations and agarose gel electrophoresis. Poly P was localized to granules randomly distributed in the sea urchin eggs, as shown by labelling with the poly-P-binding domain of Escherichia coli exopolyphosphatase. These granules were enriched using iodixanol centrifugation and shown to be acidic and to contain poly P, as determined by Acridine Orange and DAPI (4′,6′-diamidino-2-phenylindole) staining respectively. These granules also contained large amounts of calcium, sodium, magnesium, potassium and zinc, as detected by X-ray microanalysis, and bafilomycin A1-sensitive ATPase, pyrophosphatase and exopolyphosphatase activities, as well as Ca2+/H+ and Na+/H+ exchange activities, being therefore similar to acidocalcisomes described in other organisms. Calcium release from these granules induced by nigericin was associated with poly P hydrolysis. Although NAADP (nicotinic acid–adenine dinucleotide phosphate) released calcium from the granule fraction, this activity was not significantly enriched as compared with the NAADP-stimulated calcium release from homogenates and was not accompanied by poly P hydrolysis. GPN (glycyl-L-phenylalanine-naphthylamide) released calcium when added to sea urchin homogenates, but was unable to release calcium from acidocalcisome-enriched fractions, suggesting that these acidic stores are not the targets for NAADP.

Subcelluar Distribution of Myeloid-Related Protein 8 (MRP8) and MRP14 in Human Neutrophils.

Myeloid-related protein 8 (MRP8) and MRP14 are S100 family calcium binding proteins that form a heterodimer known as MRP8/14 that is present in the cytosol of neutrophils and monocytes. MRP8/14 becomes associated with endothelium at sites of monocyte and neutrophil adhesion and transmigration and induces a thrombogenic and inflammatory response by increasing the endothelial transcription of proinflamatory chemokines and adhesion molecules. MPR8/14 is the most abundant protein in neutrophil cytosol making up 30 to 60% of all cytosolic protein. However, the distribution of MPR8/MRP14 among neutrophil granules and plasma membranes in unclear and was investigated to better understand the role or MPR8/14 in acute inflammation. Three monoclonal antibodies specific for MRP8 and MPR14 were characterized, AHN-17, ANH-17.1, and 15H9, and were used to investigate the subcellar distribution of MPR8/14. The 10 and 14 kDa proteins recognized by the three antibodies were isolated by affinity chromatography with 15H9 and were separated by reverse phase high pressure liquid chromatography. N-terminal amino acid sequencing of the 10 kDa protein yielded a sequence of 29 amino acids that was identical to MRP8. The N-terminus of the 14 kDa protein was blocked, however, the amino acid sequence of two tryptic peptides were found to be identical to MRP14 at 25 of 27 amino acids. Nitrogen cavitation and density gradient separation was used to isolate neutrophil cytosol, plasma membranes, primary granules, and secondary granules. The secondary granule fraction also contains a distinct granule population termed tertiary or secretory granules. The proteins recognized by AHN-17 were isolated from each of these fractions by affinity chromatography and analyzed by SDS-PAGE under reducing conditions. MRP8 and MRP14 were located in the cytosol, plasma membrane, and secondary granule, and to a lesser degree primary granule fraction. To further determine the location of MPR-8/14 in the cellular granules, primary and secondary granules were washed and disrupted by freeze-thawing and sonication and the membranes were separated from the granule contents by centrifugation and were analyzed by immunoblotting with 15H9. Both MRP8 and MRP14 were detected in the soluble portion of the secondary granules. MPR14 was also detected in the soluble portion of primary granules and MRP8 in the plasma membrane fraction. The largest quantities of the MRP8 and MRP14 were present in cytosol and in the soluble portion of the secondary granules. MRP8 and MRP14 had a loose calcium-dependent adherence to neutrophil primary granules and plasma membranes and they were removed by washing with EGTA in a high ionic strength buffer. In conclusion MRP8/14 is located in neutrophil cytosol and in the contents of secondary granules and is loosely associated with plasma membranes and primary granules. MRP8/14 released with secondary granules by activated neutrophils likely binds to endothelium and plays an important role in acute inflammation.

The New Ketolide HMR3647 Accumulates in the Azurophil Granules of Human Polymorphonuclear Cells

ABSTRACT HMR3647 is a semisynthetic representative of a new group of drugs, the ketolides, derived from erythromycin A. Since macrolides have been shown to accumulate in human polymorphonuclear cells (PMNs), we have investigated the ability of the molecule HMR3647 to enter human PMNs as well as other cell types, such as peripheral blood mononuclear cells and cell lines of hematopoietic and nonhematopoietic origin. In these experiments, HMR3647 was compared to erythromycin A, azithromycin, clarithromycin, and roxithromycin. Our results show that HMR3647 is specifically trapped in PMNs, where it is concentrated up to 300 times. In addition, it is poorly released by these cells, 80% of the compound remaining cell associated after 2 h in fresh medium. By contrast, it is poorly internalized and quickly released by the other cell types studied. This differs from the results obtained with the macrolide molecules, which behaved similarly in the different cells studied. In addition, subcellular fractionation of PMNs allowed us to identify the intracellular compartment where HMR3647 was trapped. In PMNs, more than 75% of the molecule was recovered in the azurophil granule fraction. Similarly, in NB4 cells differentiated into PMN-like cells, almost 60% of the molecules accumulated in the azurophil granule fraction. In addition, when HMR3647 was added to disrupted PMNs, 63% accumulated in the azurophil granules. Therefore, this study shows that the ketolide HMR3647 specifically accumulates in PMN azurophil granules, thus favoring its delivery to bacteria phagocytosed in these cells.