Influence of bicarbonate on parathyroid hormone-induced changes in fluid absorption by the proximal tubule

Influence of bicarbonate on parathyroid hormone-induced changes in fluid absorption by the proximal tubule. Segments of the proximal tubule of the rabbit kidney were perfused in vitro in order to examine the influence of bicarbonate on the reduction in fluid absorption that occurs following the acute administration of parathyroid hormone (PTH). Studies were performed using either normal ultrafiltrate as perfusion fluid and normal rabbit serum as bath or low bicarbonate ultrafiltrate as perfusion fluid and low bicarbonate rabbit serum as bath. Low bicarbonate fluids were prepared by replacement of bicarbonate with chloride. In the presence of normal concentrations of bicarbonate, the addition of PTH to the bath (1 U/ml) resulted in a decrease in the fluid absorption rate (Jv)from 1.13 ±0.08 to 0.60 ± 0.04 nl/mm · min (P < 0.001) in 23 convoluted segments and from 0.64 ± 0.05 to 0.46 ± 0.05 nl/mm · min P < 0.01) in ten straight portions. Simultaneous with the PTH-induced reduction in Jv, the chloride concentration in the collected fluid changed from 119.0 ± 2.0 to 113.4 ± 1.1 mEq/liter (P < 0.01 ) in the pars convoluta and from 117.7 ± 0.6 to 114.0 ± 1.9 mEq/liter (P < 0.01 ) in the pars recta. However, there was no change in the net flux of chloride which averaged 42.58 ± 5.00pEq/mm · min during the control periods. Additional studies were performed in eight convoluted segments during perfusion on a randomized basis with low bicarbonate fluids as well as during perfusion with fluids having normal levels of bicarbonate. As before, in the presence of normal levels of bicarbonate, PTH reduced Jv from 1.16 ± 0.15 to 0.68 ± 0.07 nl/mm · min (P < 0.001) and the chloride concentration in the collected fluid ([Cl]0) from 118.6 ± 2.9 to 111.6 ± 1.3 mEq/liter (P < 0.005). Substitution of low bicarbonate fluids for normal bicarbonate fluids resulted in a decrease in Jv from 1.16 ± 0.15 to 0.74 ± 0.10 nl/mm · min (P < 0.001). In the presence of low bicarbonate fluids, the addition of PTH resulted in no further decrease in Jv (0.74 ± 0.10 vs. 0.72 ± 0.10 nl/mm · min). These data indicate that in the proximal tubule the PTH-induced reduction in fluid absorption may be mediated by changes in bicarbonate absorption

Nonimmunological alterations of glomerular filtration by s-PAF in the rat kidney

Nonimmunological alterations of glomerular filtration by s-PAF in the rat kidney. Rat kidneys were isolated and perfused with a cell-free perfusion buffer containing 4% albumin. Infusion of platelet activating factor (s-PAF) into the isolated perfused kidney caused a dose-dependent fall in renal vascular resistance (RVR): 12 ± 6% at 10nM s-PAF, 18 ± 3% at 100nM s-PAF and 20 ± 7% at 1 µM. s-PAF. Glomerular filtration rate fell by 32 ± 5% at 10nM, 38 ± 6% at 100nM, and 52 ± 10% at 1 µM. s-PAF (50nM) increased urinary protein excretion after 20 minutes. Because GFR fell to a greater extent than RVR, possible changes in glomerular permeability after s-PAF treatment were assessed morphologically using native ferritin. After s-PAF treatment (100nM), the number of ferritin particles/µm2 increased from 1.2 ± 0.9 (control) to 795 ± 69 in the glomerular basement membrane (GBM) and from 0.2 ± 0.06 (control) to 98 ± 29 in lamina rara externa (LRE). To quantitate changes in fixed anionic charges, polyethylenimine (PEI) was quantitated morphologically in GBM. No significant change between s-PAF treated and untreated kidneys was seen. s-PAF did not alter the sialoglycoprotein pattern in the perfused kidney as assessed by lysozyme staining. These results are in contrast to findings with s-PAF in vivo where in addition to increased glomerular permeability, a reduction of fixed anionic charges is seen. Thus, these results help to differentiate a dual mechanism of s-PAF: 1) a direct action of s-PAF on glomerular epithelial and vascular cells and, 2) an indirect action of s-PAF on glomerular structures via stimulation of release of inflammatory mediators from circulatory cells

Correlating liposomal adjuvant characteristics to in-vivo cell-mediated immunity using a novel Mycobacterium tuberculosis fusion protein : a multivariate analysis study

OBJECTIVE: In this study, we have used a chemometrics-based method to correlate key liposomal adjuvant attributes with in-vivo immune responses based on multivariate analysis. METHODS: The liposomal adjuvant composed of the cationic lipid dimethyldioctadecylammonium bromide (DDA) and trehalose 6,6-dibehenate (TDB) was modified with 1,2-distearoyl-sn-glycero-3-phosphocholine at a range of mol% ratios, and the main liposomal characteristics (liposome size and zeta potential) was measured along with their immunological performance as an adjuvant for the novel, postexposure fusion tuberculosis vaccine, Ag85B-ESAT-6-Rv2660c (H56 vaccine). Partial least square regression analysis was applied to correlate and cluster liposomal adjuvants particle characteristics with in-vivo derived immunological performances (IgG, IgG1, IgG2b, spleen proliferation, IL-2, IL-5, IL-6, IL-10, IFN-γ). KEY FINDINGS: While a range of factors varied in the formulations, decreasing the 1,2-distearoyl-sn-glycero-3-phosphocholine content (and subsequent zeta potential) together built the strongest variables in the model. Enhanced DDA and TDB content (and subsequent zeta potential) stimulated a response skewed towards a cell mediated immunity, with the model identifying correlations with IFN-γ, IL-2 and IL-6. CONCLUSION: This study demonstrates the application of chemometrics-based correlations and clustering, which can inform liposomal adjuvant design

Th1 immune responses can be modulated by varying dimethyldioctadecylammonium and distearoyl-sn-glycero-3-phosphocholine content in liposomal adjuvants

OBJECTIVES: Cationic liposomes of dimethyldioctadecylammonium bromide (DDA) combined with trehalose 6,6'-dibehenate (TDB) elicit strong cell-mediated and antibody immune responses; DDA facilitates antigen adsorption and presentation while TDB potentiates the immune response. To further investigate the role of DDA, DDA was replaced with the neutral lipid of distearoyl-sn-glycero-3-phosphocholine (DSPC) over a series of concentrations and these systems investigated as adjuvants for the delivery of Ag85B-ESAT-6-Rv2660c, a multistage tuberculosis vaccine. METHODS: Liposomal were prepared at a 5 : 1 DDA-TDB weight ratio and DDA content incrementally replaced with DSPC. The physicochemical characteristics were assessed (vesicle size, zeta potential and antigen loading), and the ability of these systems to act as adjuvants was considered. KEY FINDINGS: As DDA was replaced with DSPC within the liposomal formulation, the cationic nature of the vesicles decreases as does electrostatically binding of the anionic H56 antigen (Hybrid56; Ag85B-ESAT6-Rv2660c); however, only when DDA was completed replaced with DSPC did vesicle size increase significantly. T-helper 1 (Th1)-type cell-mediated immune responses reduced. This reduction in responses was attributed to the replacement of DDA with DSPC rather than the reduction in DDA dose concentration within the formulation. CONCLUSION: These results suggest Th1 responses can be controlled by tailoring the DDA/DSPC ratio within the liposomal adjuvant system

Th1 immune responses can be modulated by varying dimethyldioctadecylammonium and distearoyl-sn-glycero-3-phosphocholine content in liposomal adjuvants

Objectives Cationic liposomes of dimethyldioctadecylammonium bromide (DDA) combined with trehalose 6,6′-dibehenate (TDB) elicit strong cell-mediated and antibody immune responses; DDA facilitates antigen adsorption and presentation while TDB potentiates the immune response. To further investigate the role of DDA, DDA was replaced with the neutral lipid of distearoyl-sn- glycero-3-phosphocholine (DSPC) over a series of concentrations and these systems investigated as adjuvants for the delivery of Ag85B-ESAT-6-Rv2660c, a multistage tuberculosis vaccine. Methods Liposomal were prepared at a 5: 1 DDA-TDB weight ratio and DDA content incrementally replaced with DSPC. The physicochemical characteristics were assessed (vesicle size, zeta potential and antigen loading), and the ability of these systems to act as adjuvants was considered. Key findings As DDA was replaced with DSPC within the liposomal formulation, the cationic nature of the vesicles decreases as does electrostatically binding of the anionic H56 antigen (Hybrid56; Ag85B-ESAT6-Rv2660c); however, only when DDA was completed replaced with DSPC did vesicle size increase significantly. T-helper 1 (Th1)-type cell-mediated immune responses reduced. This reduction in responses was attributed to the replacement of DDA with DSPC rather than the reduction in DDA dose concentration within the formulation. Conclusion These results suggest Th1 responses can be controlled by tailoring the DDA/DSPC ratio within the liposomal adjuvant system. © 2013 Royal Pharmaceutical Society

Numerical Solution of the S‐Limit Schrödinger Equation

Numerical solutions to the S‐limit Schrödinger equation have been obtained for He and Li^+. Using these the energy and the expectation values 〈Σr_i〉 and 〈Σr_(i^2)〉 were calculated and compared to the radial configuration interaction values. The results demonstrate that the direct numerical solution of many partial differential equations in chemical physics can be accomplished in a practical and straightforward manner