In situ remineralizing effect of fluoride varnishes containing sodium trimetaphosphate

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Synergistic effect of fluoride and sodium hexametaphosphate in toothpaste on enamel demineralization in situ

To evaluate the effect of a fluoride dentifrice containing sodium hexametaphosphate (HMP) on enamel demineralization in situ. This double-blind and cross-over study consisted of 3 phases (7 days each) in which 12 volunteers wore intraoral appliances containing four enamel bovine blocks. Specimens were treated (3×/day) with placebo (no F or HMP), 1100ppm F (1100F) and 1100F plus HMP1% (1100F-HMP1%) toothpastes, and the cariogenic challenge was performed using a 30% sucrose solution (6×/day). Final surface hardness, the percentage of surface hardness loss (%SH), the integrated loss of subsurface hardness (ΔKHN), as well as enamel calcium (Ca), phosphorus (P) and firmly-bound fluoride (F) were determined. Also, biofilm formed on the blocks were analyzed for F, Ca, P and insoluble extracellular polysaccharide (EPS) concentrations. Data were submitted 1-way ANOVA, followed by Student-Newman-Keuls' test (p<0.05). 1100F-HMP1% promoted the lowest %SH and ΔKHN among all groups (p<0.001). The addition of HMP1% to 1100F did not enhance enamel F uptake, but significantly increased enamel Ca concentrations (p<0.001). Similar EPS concentrations were seen for 1100F-HMP1% and 1100F groups (p>0.05). All the groups were supersaturated with respect to HA. However, only 1100F-HMP1% group was supersaturated with respect to CaF2 (p<0.05). The ionic activities of F(-), CaF(+) and HF(0) for the 1100F-HMP1% group were the highest among all groups (p<0.001). The addition of HMP1% to a conventional toothpaste significantly reduces enamel demineralization in situ when compared to 1100F. This dentifrice could be a viable alternative to patients at high risk of caries.

Effectiveness of a toothpaste with low fluoride content combined with trimetaphosphate on dental biofilm and enamel demineralization in situ

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

In vitro effect of low-fluoride toothpastes containing sodium trimetaphosphate on enamel erosion

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Adição de plasma bovino em salmouras para injeção de coxão duro bovino (m. Biceps femoris) e seus efeitos no pH e na carga microbiana de bifes cozidos embalados à vácuo e mantidos sob refrigeração

The physical (pH) and microbiological (psychotrofi c microorganisms and lactic bacteria) characteristics of beef outside round (m. Biceps femoris) injected (15%) with brines free of polyphosphates containing and sodium lactate or sodium lactate and sodium diacetate and liquid bovine plasma (PLL and PLO) or dehydrated bovine plasma (PDL, PDO) were evaluated along with beef cuts injected with brines free from plasma, but containing polyphosphates and bacteriostatic agents (CL and CO) and non injected beef cuts (IN), comprising seven treatments of cooked and vacuum packaged beef steaks stored under refrigeration (6ºC) during 43 days. No differences in pH were detected among raw or cooked injected treatments, although IN showed lower pH value in raw beef cuts. The addition of liquid or dehydrated bovine plasma did not affect the microbial load after whole muscles pasteurization, but increased the bacterial counts in cooked beef steaks during refrigerated storage, comparing to treatments with no plasma addition (CL and CO). The storage temperature (6ºC), usually found during commercialization of meat increased the microorganisms growth rate affecting the microbiological quality, especially when plasma was added to the brine.

Calcium-dependent switching of the specificity of phosphoinositide binding to synaptotagmin

The synaptic vesicle membrane protein synaptotagmin (tagmin) is essential for fast, calcium-dependent, neurotransmitter release and is likely to be the calcium sensor for exocytosis, because of its many calcium-dependent properties. Polyphosphoinositides are needed for exocytosis, but it has not been known why. We now provide a possible connection between these observations with the finding that the C2B domain of tagmin I binds phosphatidylinositol-4,5-bisphosphate (PIns-4,5-P2), its isomer phosphatidylinositol-3,4-bisphosphate and phosphatidylinositol-3,4,5-trisphosphate (PIns-3,4,5-P3). Calcium ions switch the specificity of this binding from PIns-3,4,5-P3 (at calcium concentrations found in resting nerve terminals) to PIns-4,5-P2 (at concentration of calcium required for transmitter release). Inositol polyphosphates, known blockers of neurotransmitter release, inhibit the binding of both PIns-4,5-P2 and PIns-3,4,5-P3 to tagmin. Our findings imply that tagmin may operate as a bimodal calcium sensor, switching bound lipids during exocytosis. This connection to polyphosphoinositides, compounds whose levels are physiologically regulated, could be important for long-term memory and learning.

Unblocking of chain-terminated primer by HIV-1 reverse transcriptase through a nucleotide-dependent mechanism

HIV-1 replication is inhibited by the incorporation of chain-terminating nucleotides at the 3′ end of the growing DNA chain. Here we show a nucleotide-dependent reaction catalyzed by HIV-1 reverse transcriptase that can efficiently remove the chain-terminating residue, yielding an extendible primer terminus. Radioactively labeled 3′-terminal residue from the primer can be transferred into a product that is resistant to calf intestinal alkaline phosphatase and sensitive to cleavage by snake venom phosphodiesterase. The products formed from different nucleotide substrates have unique electrophoretic migrations and have been identified as dinucleoside tri- or tetraphosphates. The reaction is inhibited by dNTPs that are complementary to the next position on the template (Ki ≈ 5 μM), suggesting competition between dinucleoside polyphosphate synthesis and DNA polymerization. Dinucleoside polyphosphate synthesis was inhibited by an HIV-1 specific non-nucleoside inhibitor and was absent in mutant HIV-1 reverse transcriptase deficient in polymerase activity, indicating that this activity requires a functional polymerase active site. We suggest that dinucleoside polyphosphate synthesis occurs by transfer of the 3′ nucleotide from the primer to the pyrophosphate moiety in the nucleoside di- or triphosphate substrate through a mechanism analogous to pyrophosphorolysis. Unlike pyrophosphorolysis, however, the reaction is nucleotide-dependent, is resistant to pyrophosphatase, and produces dinucleoside polyphosphates. Because it occurs at physiological concentrations of ribonucleoside triphosphates, this reaction may determine the in vivo activity of many nucleoside antiretroviral drugs.

Identification of Inositol 1,3,4-Trisphosphate 5-Kinase and Inositol 1,3,4,5-Tetrakisphosphate 6-Kinase in Immature Soybean Seeds

In extracts of immature soybean (Glycine max [L.] Merr.) seeds inositol tetrakisphosphate was formed from [3H]inositol 1,3,4-trisphosphate but not from [3H]inositol 1,4,5-trisphosphate. Inositol 1,3,4-trisphosphate kinase was purified to a specific activity of 3.55 min−1 mg−1 by polyethylenimine clarification and anion-exchange chromatography. The partially purified enzyme converted [3H]inositol 1,3,4-trisphosphate to inositol 1,3,4,5-tetrakisphosphate as the major product and inositol 1,3,4,6- and/or 1,2,3,4-tetrakisphosphate as the minor product. Subsequent experiments revealed a separate inositol 1,3,4,5-tetrakisphosphate 6-kinase activity, which could link these enzymes to inositol hexakisphosphate synthesis via the previously reported inositol 1,3,4,5,6-pentakisphosphate 2-kinase. The apparent Km values for inositol 1,3,4-trisphosphate kinase were 200 ± 0 nm for inositol 1,3,4-trisphosphate and 171 ± 4 μm for ATP, and the reaction was not reversible. The kinetics were such that no activity could be detected using unlabeled inositol 1,3,4-trisphosphate and [γ-32P]ATP, which suggested that other kinases may have been observed when less purified fractions were incubated with radiolabeled ATP. Inositol 1,3,4-trisphosphate kinase was nonspecifically inhibited more than 80% by various inositol polyphosphates at a concentration of 100 μm.

Identification of a novel inositol phosphate recognition site: specific [3H]inositol hexakisphosphate binding to brain regions and cerebellar membranes

[3H]Inositol hexakisphosphate (InsP6) binds with a heterogeneous distribution to frozen sections of unfixed rat brain and is displaced by unlabelled InsP6. The pattern of binding correlates with binding to neuronal cell bodies. [3H]InsP6 binding to cerebellar membranes has been further characterised, is reversible, and saturable, and exhibits high specificity for inositol polyphosphates. The IC50 for competition by unlabelled InsP6 is approximately 100nM, whereas inositol 1,3,4,5,6 pentakisphosphate (Ins(13456)P5), inositol 1,3,4,5 tetrakisphosphate (Ins(1345)P4), and inositol 1,4,5 trisphosphate (Ins(145)P3) bind with an affinity at least one order of magnitude lower. [3H]InsP6 binding is clearly distinct from previously characterised Ins(145)P3 (ref. 1, 2) and Ins(1345)P4 (ref. 3) binding, both in terms of pharmacology and brain distribution.