Molecular cloning and characterization of an inorganic phosphate transporter protein encoding gene in Polyporus umbellatus / 中草药

Objective To clone an inorganic phosphate transporter (PiT) gene from Polyporus umbellatus and perform the bioinformatics and expression mode analysis. Methods Using RT-PCR.to clone the full-length cDNA of PiT. The characteristics of physiochemical properties, conserved domains, signal peptide and transmembrane domain of the predicted PiT protein were determined by using bioinformatic tools. Results A inorganic phosphate transporter (PiT) gene (NCBI: KU179154), designated as PuPiT, was cloned from Polyporus umbellatus sclerotia by RT-PCR. The full open reading frame cDNA sequence of PuPiT was 1 590 bp, encoding a putative PiT protein with 530 amino acids with a molecular weight of 57 552, and a theoretical pI of 6.82. The amino acids possess 12 membrane-spanning domains. Phylogenetic tree analysis indicated that PuPiT had the highest similarity with PiT from Moniliophthora rorer, and had high similarity with Moniliophthora roreri, Laccaria bicolor, and Heterobasidion irregulare. Quantitative real-time PCR showed that PuPiT expressed in both the symbiotic part and non-symbiotic part. Meanwhile, the expression of PuPiT in the symbiotic part was significantly up-regulated, about 12 times more than that in the non-symbiotic part. This result showed that PuPiT might play an important role in the Pi accumulating. Conclusion Molecular cloning and characterization of the novel PuPiT gene will be useful for further functional determination of the gene involving in phosphorus translocation regulation and symbiotic process.

Research and Application of Analytical Technique onδ18Op of Inorganic Phosphate in Soil / 分析化学

Analytical technique on oxygen isotope composition of phosphate in soil has important applications in biogeochemical research. As the composition of the soil is complex with multiple oxygen sources, a liable phosphate enrichment and purification technology is important for phosphate oxygen isotope research. Here we presented a protocol on the analysis of the δ18 Op of inorganic phosphate in soil, and phosphate was purified with multiple mineral precipitations and finally precipitated as silver phosphate. The efficiency of NH4 OH purification method was examined and the purity of the product was verified through XRD and element analysis. The result showed that treatment of the Ag3 PO4 sample by recrystallizing with ammonia could remove the impurity efficiently without significantly modifying oxygen isotope compositions. The validity of sample processing and reliability of isotope analyses were confirmed by processing a KH2 PO4 standard in parallel with soil P sample. The analytical result of three soil types indicated that this method provided good precision of 0. 2‰. The differences of δ18 Op between three soils groups are significant ( From 15. 2‰ to 19. 5‰) and can be used to tracing P sources.

Effect of different 1 percent chlorhexidine varnish regimens on biochemical composition of the dental biofilm / Efeito de diferentes posologias do verniz de clorexidina a 1 por cento na composição bioquímica do biofilme dentário

PURPOSE: This investigation evaluated in a randomized controlled study the effect of different 1 percent chlorhexidine varnish (1 percentCHX-V) regimens on biochemical composition of the dental biofilm. METHODS: Subjects with mutans streptococci > 10(5) CFU/mL saliva, were allocated into 4 groups: A - single 1 percent CHX-V application; B - 1 percent CHX-V was applied once daily on 3 consecutive days; C - 1 percent CHX-V was applied 3 times with an interval of 4 days between each application; and D - placebo varnish was applied once daily on 3 consecutive days. Dental biofilm samples were collected at baseline and 1, 4, and 8 weeks after the final varnish application and evaluated for inorganic phosphate and insoluble polysaccharide concentrations. RESULTS: No difference was observed between all groups during the different experimental periods in relation to biochemical composition. After 1 week, a significant increase was observed in inorganic phosphate concentration, returning to baseline values after 4 weeks. All groups showed a significant decrease in insoluble polysaccharide concentration for more than 8 weeks after treatment. CONCLUSION: Varnish application resulted in modification of the dental biofilm toward a less cariogenic one.

OBJETIVO: Este estudo avaliou o efeito de diferentes posologias do verniz de clorexidina a 1 por cento (V-CHX1 por cento) na composição bioquímica do biofilme dental. METODOLOGIA: Voluntários com níveis de Streptococcus mutans > 10(5) UFC/mL saliva, foram divididos em 4 grupos: A - uma única aplicação do V-CHX1 por cento; B - aplicações de 1x/dia do V-CHX1 por cento durante 3 dias consecutivos; C - 3 aplicações de 1x/dia do V-CHX1 por cento com intervalos de 4 dias entre elas; D - aplicações de 1x/dia do verniz placebo durante 3 dias consecutivos. Amostras de biofilme dental foram coletadas, no início e após 1, 4 e 8 semanas das aplicações dos vernizes, e analisadas quanto às concentrações de fosfato inorgânico (Pi) e polissacarídeos insolúveis (PI). RESULTADOS: Não foram observadas diferenças, em relação à composição bioquímica, entre os grupos durante os diferentes períodos experimentais. Foi observado, após 1 semana, um aumento significativo nas concentrações de Pi, retornando aos valores iniciais após 4 semanas. Todos os grupos apresentaram uma redução significativa na concentração de PI por mais de 8 semanas após os tratamentos. CONCLUSÃO: A aplicação do verniz resultou em uma modificação do biofilme dentário tornando-o menos cariogênico.

Responses of parathyroid gland, C cells, and plasma calcium and inorganic phosphate levels in rat to sub-lethal heroin administration.

In order to record the effects of heroin on plasma calcium (Ca) and inorganic phosphate (Pi) levels as well as parathyroid gland and C cells, two sub-lethal doses (0.50 LD50 and 0.75 LD50) of the drug were administered intramuscularly in Rattus norvegicus for 30 days. Plasma Ca level of control rats ranged between 9.53±0.32 - 9.88±0.22 mg 100 ml-1 while plasma Pi concentration fluctuated between 4.55±0.18 - 4.71±0.24 mg 100 ml-1. Sub-lethal heroin administration induced progressive increase in plasma Ca level during the first seven days (p <0.001), thereafter the level declined on day 15 and 30. However, plasma Pi level of the heroin-treated rats registered increase with the peak value (p <0.001) on day 30. The treatment elicited degenerative changes in parathyroid gland as evident by cytoplamic vacuolization, presence of more pycnotic nuclei and occurrence of patchy areas among the chief cells. Degenerative changes were also noticed in cristae of mitochondria, Golgi complex and endoplasmic reticulum. There was decrease in chromatin material in the nucleus and loss of hormone granules in the cytoplasm. Oxyphil cells of the heroin-treated rat depicted dilation of endoplasmic reticulum and damaged cristae. Sub-lethal heroin administration in the rat for 30 days induced dilation in endoplasmic reticulum and loss of secretory granules in C cells.

Low dietary inorganic phosphate affects the lung growth of developing mice

Inorganic phosphate (Pi) plays a critical role in diverse cellular functions, and regulating the Pi balance is accomplished by sodium-dependent Pi co-transporter (NPT). Pulmonary NPT has recently been identified in mammalian lungs. However, to date, many of the studies that have involved Pi have mainly focused on its effect on bone and kidney. Therefore, current study was performed to discover the potential effects of low Pi on the lung of developing transgenic mice expressing the renilla/firefly luciferase dual reporter gene. Two-weeks old male mice divided into 2 groups and these groups were fed either a low PI diet or a normal control diet (normal: 0.5% Pi, low: 0.1% Pi) for 4 weeks. After 4 weeks of the diet, all the mice were sacrificed. Their lungs were harvested and analyzed by performing luciferase assay, Western blotting, kinase assay and immunohistochemistry. Our results demonstrate that low Pi affects the lungs of developing mice by disturbing protein translation, the cell cycle and the expression of fibroblast growth factor-2. These results suggest that optimally regulating Pi consumption may be important to maintain health.

The Effect of Saponin on the Na+ - K+ Activated ATPase of Cattle Lens Epithelial Cell

The effect of saponin on the Na+ - K+ ATPase of anterior capsule and epithelium of cattle lens has been investigated. The experiment also designed to determine the cation of saponin on the Na+ - K+ ATPase activity in anterior capsule and epithelium of cattle lens. The following results are observed. 1. The Na+ - K+ ATPase activity of cattle lens epithelial cell membrane is inhibited by low concentration of saponin, but increased by high concentration. The strongest activity showed at the dose of 50mg% saponin concentration. 2. The activating effect of saponin on the cattle epithelial cell membrane Na+ - K+ ATPase activity is inhibited by ouabain, but the stimulation of the Mg++ ATPase by high concentration of saponin is not inhibited by ouabain. 3. The activity ratio of Na+ - K+ ATPase by high concentration of saponin is increased by raising the sodium concentration but decreased by raising the potassium concentration. 4. The Na+ - K+ ATPase activity is increased by small amount of calcium but completely inhibited by over 5mM concentration of calcium.

Effect of Carteolol Hydrochloride on Sodium-Potassium Activated ATPase in Cattle Lena Epithelial Cell Membrane

The effect of Carteolol hydrocholride on the Na-K ATPase of anterior capsule and epithelium of cattle lens has been investigated. The experiments were also designed to determine the action of Carteolol hydrochloride on the Na-K ATPase activity in anterior capsule and epithelium of cattle lens. The following results were observed. 1. The predominent location of Na-K ATPase was located in the epithelim and NaK ATPase was closely related with active trasport system of sodium and potassium in the lens epithelium. 2. The activity of Na-K ATPase of cattle lens epithelium was alomst totally inhibited by Ouabain. 3. Beta-adrenergic blocking agent(Carteolol hydrochloride) in the epithelium of cattle lens was shown to be actively transported by the ATPase. 4. The Na-K ATPase activity of cattle lens epithelium was inhibited by Carteolol hydrochloride.