Distribution Of Plasma Proteins Research Articles

Normative Data of Beta-2-Microglobulin and Plasma Proteins from an Agrarian Indian Community By a Door-to-Door Survey: Results of Simple (IMAGe-002B) Study

Introduction: Most diagnostic laboratories in India follow the reference intervals given in Western literature whose Caucasian reference population is entirely different from the Indian ethnic population (viz., dietary habits, lifestyle, socioeconomic status, ethnicity, and environmental factors). Objectives: To collate normative data of beta-2-microglobulin (β2M) and plasma proteins from an otherwise healthy population from an agrarian Indian Community. The secondary objectives were to evaluate the correlation of β2M levels with age, gender, and BMI of these individuals. Methods: A cross-sectional study was conducted in an agrarian village in India with a 24.76Km perimeter and 4588.3 hectares with an 89.2% population coverage for adults (>45y) spending 12096 man-hours. This was a part of SIMPLe (Screening Intervention for Myeloma and Prevention of Lifestyle diseases) study registered vide CTRI/2023/03/051220 after obtaining informed consent. Demographic details, medical history, and blood samples were collected after a thorough medical examination to ascertain the normal health status of the study participants. Hemogram, biochemistry (Liver and renal function tests), β2M, and serum protein electrophoresis (SPEP) were performed in all individuals. Only individuals with normal renal and liver function tests were included in the final analysis. Bivariate analysis (Fit Y by X) was done to study the correlations. Any p value < 0.05 was considered significant. The data was analyzed using JMP ver. 16.0.0. Results: A total of 850 individuals were included in this community-based door-to-door survey. The mean age of the study population was 57.6±10.5y, with a slight female predominance (56.3%). The distribution of β2M and various plasma proteins is mentioned in Table 1. The β2M significantly differed between males and females (χ 2-8.58; dF-1; p-0.0034). On bivariate analysis, β2M was significantly associated with age (correlation coefficient: 0.31; p<0.0001), and BMI (correlation coefficient -0.07; p-0.023). Conclusion: The data obtained from our studies would act as the first step in collating normative data for β2M and plasma proteins from Indian community.

Assessing Dose-Exposure–Response Relationships of Miltefosine in Adults and Children using Physiologically-Based Pharmacokinetic Modeling Approach

ObjectivesMiltefosine is the first and only oral medication to be successfully utilized as an antileishmanial agent. However, the drug is associated with differences in exposure patterns and cure rates among different population groups e.g. ethnicity and age (i.e., children v adults) in clinical trials. In this work, mechanistic population physiologically-based pharmacokinetic (PBPK) models have been developed to study the dose-exposure–response relationship of miltefosine in in silico clinical trials and evaluate the differences in population groups, particularly children and adults.MethodsThe Simcyp population pharmacokinetics platform was employed to predict miltefosine exposure in plasma and peripheral blood mononuclear cells (PBMCs) in a virtual population under different dosing regimens. The cure rate of a simulation was based on the percentage of number of the individual virtual subjects with AUCd0-28 > 535 µg⋅day/mL in the virtual population.ResultsIt is shown that both adult and paediatric PBPK models of miltefosine can be developed to predict the PK data of the clinical trials accurately. There was no significant difference in the predicted dose-exposure–response of the miltefosine treatment for different simulated ethnicities under the same dose regime and the dose-selection strategies determined the clinical outcome of the miltefosine treatment. A lower cure rate of the miltefosine treatment in paediatrics was predicted because a lower exposure of miltefosine was simulated in virtual paediatric in comparison with adult virtual populations when they received the same dose of the treatment.ConclusionsThe mechanistic PBPK model suggested that the higher fraction of unbound miltefosine in plasma was responsible for a higher probability of failure in paediatrics because of the difference in the distribution of plasma proteins between adults and paediatrics. The developed PBPK models could be used to determine an optimal miltefosine dose regime in future clinical trials.Graphical

Clinical Pharmacokinetic Assessment of Kratom (Mitragyna speciosa), a Botanical Product with Opioid-like Effects, in Healthy Adult Participants.

Increasing use of the botanical kratom to self-manage opioid withdrawal and pain has led to increased kratom-linked overdose deaths. Despite these serious safety concerns, rigorous fundamental pharmacokinetic knowledge of kratom in humans remains lacking. We assessed the pharmacokinetics of a single low dose (2 g) of a well-characterized kratom product administered orally to six healthy participants. Median concentration-time profiles for the kratom alkaloids examined were best described by a two-compartment model with central elimination. Pronounced pharmacokinetic differences between alkaloids with the 3S configuration (mitragynine, speciogynine, paynantheine) and alkaloids with the 3R configuration (mitraciliatine, speciociliatine, isopaynantheine) were attributed to differences in apparent intercompartmental distribution clearance, volumes of distribution, and clearance. Based on noncompartmental analysis of individual concentration-time profiles, the 3S alkaloids exhibited a shorter median time to maximum concentration (1–2 vs. 2.5–4.5 h), lower area under the plasma concentration-time curve (430–490 vs. 794–5120 nM × h), longer terminal half-life (24–45 vs. ~12–18 h), and higher apparent volume of distribution during the terminal phase (960–12,700 vs. ~46–130 L) compared to the 3R alkaloids. Follow-up mechanistic in vitro studies suggested differential hepatic/intestinal metabolism, plasma protein binding, blood-to-plasma partitioning, and/or distribution coefficients may explain the pharmacokinetic differences between the two alkaloid types. This first comprehensive pharmacokinetic characterization of kratom alkaloids in humans provides the foundation for further research to establish safety and effectiveness of this emerging botanical product.

Sterol Constituents of Sea Fan (Gorgonia mariae) as Potential Candidates of MPro Protein SARS-CoV-2 Inhibitor: in silico Study

Ethnobotanically, sea fan was a group of gorgonian coral that have used the Maluku people as medicinal ingredients with secondary metabolites containing sterols, terpenoids, and alkaloids that have anti-inflammatory, anti-viral, anti-cancer, analgesic, gastroprotective, anti-bacterial, anti-cancer, and anti-fouling agents. However, the effectiveness of sterols as anti-viral SARS-CoV-2 has not been reported, so research was needed. The initial stage of targeting (SwissTargetPrediction), binding affinity (Autodock Tools 4.2), and amino acid interactions (Discovery Studio 2016 Client®). The route of administration, pharmacokinetic properties, and acute oral toxicity (LD50) were predicted by Lipinski's rule of five, pre-ADMET, and ProTox-II. The results of target class obtained probability of 10.6% (4,24-dimethyl cholesta-7,22-dien-3ß-ol and 4,24-dimethyl-22-dehydro-cholestanol) and 11, 8% (dinosterol). Binding affinity (ΔG, kcal/mol and Ki, nM) potentially 4,24-dimethyl cholesta-7,22-dien-3ß-ol (-9.90; 55.13) > dinosterol (-9.77; 68.66) > 4,24-dimethyl-22-dehydro-cholestanol (-9.48; 113.33), respectively with the crucial amino acid, Asp187. The test compound has a log P value > 5, so solubility must be considered. Pre-ADMET showed an excellent disposition as a drug and was not mutagenic and carcinogenic. However, the distribution of plasma proteins and the dose of LD50 need to be considered. Thus, sea fan sterols have potential as MPro protein inhibitors.

Bacterial serum opacity factor catalyzes the discordant distribution of plasma proteins: Analysis by asymmetric flow flow-field fractionation (AF4)

Background and Aims: Serum opacity factor (SOF), a highly stable bacterial protein, converts HDL to three products, lipid-free APOA1, lipoprotein remnants (neo HDL), and large cholesteryl ester rich microemulsions (CERM). SOF has potential cardioprotective activities: both neo HDL and APOA1 support macrophage-free cholesterol efflux, and low-dose SOF in mice lowers plasma cholesterol (>50%) via the APOE-mediated targeting of CERM to hepatic receptors for disposal. Previous studies characterized the SOF reaction vs.

Протеїнограма крові тварин в стані штучного гіпобіозу за ешерихіозу

The features of fractional distribution of plasma proteins in rats with Escherichia coli at different stages of infection are established. It was studied the effect of hypobiosis on the proteinogramma under and after artificial sleep too. Artificial hypobiosis is important for therapeutic use at bacterial infection and proteinogramma is informatively significant as a diagnostic element of the functional changes provoked by a hypobiosis. We used the nephelometric method. Experiments were performed on young rats with Escherichia coli, in a state of hypobiosis and one day after leaving it. It was established the infection provoked changes in plasma proteins such as increasing globulins and decreasing albumin at the beginning of the disease and the opposite trend during the period of total depletion. The introduction of sick animals into a state of hypobiosis causes an increase in nonspecific resistance of the organism level due to an increase in the content of γ-globulin fraction and a decrease in the level of albumin, which increases the survival of animals after removal from hypobiosis. Indicators of the proteinogram of blood of animals that were introduced into the state of hypobiosis at the initial stage of the disease and the stage of progressive symptoms return to control values on the first day after the elimination of hypobiosis factors. Further research should be aimed at a comprehensive study of functional changes in the body according to the proposed model.

Effects of paracetamol (acetaminophen) on gene expression and permeability properties of the rat placenta and fetal brain.

Background: Paracetamol (acetaminophen) is widely used in pregnancy and generally regarded as "safe" by regulatory authorities. Methods: Clinically relevant doses of paracetamol were administered intraperitoneally to pregnant rats twice daily from embryonic day E15 to 19 (chronic) or as a single dose at E19 (acute). Control samples were from un-treated age-matched animals. At E19, rats were anaesthetised, administered a final paracetamol dose, uteruses were opened and fetuses exposed for sample collection. For RNA sequencing, placentas and fetal brains were removed and flash frozen. Fetal and maternal plasma and cerebrospinal fluid were assayed for α-fetoprotein and interleukin 1β (IL1β). Brains were fixed and examined (immunohistochemistry) for plasma protein distribution. Placental permeability to a small molecule ( 14C-sucrose) was tested by injection into either mother or individual fetuses; fetal and maternal blood was sampled at regular intervals to 90 minutes. Results: RNA sequencing revealed a large number of genes up- or down-regulated in placentas from acutely or chronically treated animals compared to controls. Most notable was down-regulation of three acute phase plasma proteins (α-fetoprotein, transferrin, transthyretin) in acute and especially chronic experiments and marked up-regulation of immune-related genes, particularly cytokines, again especially in chronically treated dams. IL1β increased in plasma of most fetuses from treated dams but to variable levels and no IL1β was detectable in plasma of control fetuses or any of the dams. Increased placental permeability appeared to be only from fetus to mother for both 14C-sucrose and α-fetoprotein, but not in the reverse direction. In the fetal brain, gene regulatory changes were less prominent than in the placenta of treated fetuses and did not involve inflammatory-related genes; there was no evidence of increased blood-brain barrier permeability. Conclusion: Results suggest that paracetamol may induce an immune-inflammatory-like response in placenta and more caution should be exercised in use of paracetamol in pregnancy.

Effects of paracetamol (acetaminophen) on gene expression and permeability properties of the rat placenta and fetal brain

Background: Paracetamol (acetaminophen) is widely used in pregnancy and generally regarded as “safe” by regulatory authorities. Methods: Clinically relevant doses of paracetamol were administered intraperitoneally to pregnant rats twice daily from embryonic day E15 to 19 (chronic) or as a single dose at E19 (acute). Control samples were from un-treated age-matched animals. At E19, rats were anaesthetised, administered a final paracetamol dose, uteruses were opened and fetuses exposed for sample collection. For RNA sequencing, placentas and fetal brains were removed and flash frozen. Fetal and maternal plasma and cerebrospinal fluid were assayed for α-fetoprotein and interleukin 1β (IL1β). Brains were fixed and examined (immunohistochemistry) for plasma protein distribution. Placental permeability to a small molecule ( 14C-sucrose) was tested by injection into either mother or individual fetuses; fetal and maternal blood was sampled at regular intervals to 90 minutes. Results: RNA sequencing revealed a large number of genes up- or down-regulated in placentas from acutely or chronically treated animals compared to controls. Most notable was down-regulation of three acute phase plasma proteins (α-fetoprotein, transferrin, transthyretin) in acute and especially chronic experiments and marked up-regulation of immune-related genes, particularly cytokines, again especially in chronically treated dams. IL1β increased in plasma of most fetuses from treated dams but to variable levels and no IL1β was detectable in plasma of control fetuses or any of the dams. Increased placental permeability appeared to be only from fetus to mother for both 14C-sucrose and α-fetoprotein, but not in the reverse direction. In the fetal brain, gene regulatory changes were less prominent than in the placenta of treated fetuses and did not involve inflammatory-related genes; there was no evidence of increased blood-brain barrier permeability. Conclusion: Results suggest that paracetamol may induce an immune-inflammatory-like response in placenta and more caution should be exercised in use of paracetamol in pregnancy.

Effects of paracetamol (acetaminophen) on gene expression and permeability properties of the rat placenta and fetal brain.

Background: Paracetamol (acetaminophen) is widely used in pregnancy and generally regarded as "safe" by regulatory authorities. Methods: Clinically relevant doses of paracetamol were administered intraperitoneally to pregnant rats twice daily from embryonic day E15 to 19 (chronic) or as a single dose at E19 (acute). Control samples were from un-treated age-matched animals. At E19, rats were anaesthetised, administered a final paracetamol dose, uteruses were opened and fetuses exposed for sample collection. For RNA sequencing, placentas and fetal brains were removed and flash frozen. Fetal and maternal plasma and cerebrospinal fluid were assayed for ⍺-fetoprotein and interleukin 1β (IL1β). Brains were fixed and examined (immunohistochemistry) for plasma protein distribution. Placental permeability to a small molecule ( 14C-sucrose) was tested by injection into either mother or individual fetuses; fetal and maternal blood was sampled at regular intervals to 90 minutes. Results: RNA sequencing revealed a large number of genes up- or down-regulated in placentas from acutely or chronically treated animals compared to controls. Most notable was down-regulation of three acute phase plasma proteins (⍺-fetoprotein, transferrin, transthyretin) in acute and especially chronic experiments and marked up-regulation of immune-related genes, particularly cytokines, again especially in chronically treated dams. IL1β increased in plasma of most fetuses from treated dams but to variable levels and no IL1β was detectable in plasma of control fetuses or any of the dams. Increased placental permeability appeared to be only from fetus to mother for both 14C-sucrose and ⍺-fetoprotein, but not in the reverse direction. In the fetal brain, gene regulatory changes were less prominent than in the placenta of treated fetuses and did not involve inflammatory-related genes; there was no evidence of increased blood-brain barrier permeability. Conclusion: Results suggest that paracetamol may induce an immune-inflammatory-like response in placenta and more caution should be exercised in use of paracetamol in pregnancy.

Preliminary Evaluation of Seminal Plasma Proteins and Immunoreactivity of Nerve Growth Factor as Indicative of an Ovulation Inducing Factor in Odontocetes

In the seminal plasma of terrestrial mammalian species known as induced (e.g., camels) and spontaneous (e.g., cattle) ovulators, an ovulation-inducing factor (OIF) with a protein structure similar to beta-nerve growth factor (β-NGF) has been identified. Detection of an OIF/NGF in the seminal plasma of cetaceans would have both basic and applied implications in reproductive biology and conservation management programs. A preliminary evaluation was conducted to characterize the distribution and abundance of seminal plasma proteins in aquarium-based belugas and a Pacific white-sided and bottlenose dolphin. Initially, SDS-PAGE was used with 50 μg of total protein for separation; thereafter, Western immunoblot was used with anti-NGF. In addition to odontocete seminal plasma, a purified fraction of llama seminal plasma (100 ng protein) and an extract of mouse brain (20 μg total protein) were included as positive controls for NGF. Within the two belugas, visual inspection of the protein bands indicated similar distribution and intensity. However, among the belugas and Pacific white-sided and bottlenose dolphins there was more diversity than similarity in the distribution and abundance of seminal plasma proteins. While immunoreactivity of NGF was distinctly evident in the llama and mouse positive controls, there was no visual reactivity in any of the odontocete samples. These preliminary results provide novel information indicating more homogeneity within and heterogeneity among seminal plasma proteins of ondentocetes. Although NGF was not immunologically detected, future studies are required to address the apparent limitations of immuno-detection of NGF, especially if the post-translational form of β-NGF is in low abundance in the seminal plasma of belugas and Pacific white-sided and bottlenose dolphins.

Chromatographic and Computational Assessment of Potential Biological Activity of N-(Substituted Phenyl)- 2-Chloroacetamides Applying Multivariate Methods

The lipophilicity of the newly synthesized N-(substituted phenyl)-2-chloroacetamides, as the most commonly used molecular descriptor of a potential biological activity was the subject of research. The lipophilicity of the tested derivatives was determined by applying the reversed-phase thin-layer chromatography (RPTLC18F254s) in mixtures of water and one protic (n-propanol) and one aprotic (acetonitrile) solvent, as well as mathematically. The effects of the substituent on the lipophilicity of chloroacetamides were discussed. The obtained chromatographic retention constant, , of examined chloroacetamides was correlated with the standard measure of lipophilicity, log P, and with pharmacokinetic predictors such as human effective permeability in jejunum, Peff, plasma protein binding, PPB, and distribution through blood-brain barrier, log BBB, using linear regression analysis and two multivariate methods, cluster analysis and principal component analysis. All the applied methods gave very similar results, and all the procedures also confirmed the fact that determined by RPTLC could be used as a descriptor to estimate the potential biological activity of N-(substituted phenyl)-2-chloroacetamides.

Seasonal dynamics in capillary filtration of plasma proteins in eastern redfins of the genus Tribolodon (Cyprinidae)

A wide range of the osmotic gradient created by plasma and tissue fluid proteins in relation to the capillary wall in redfins of the genus Tribolodon during an annual cycle was revealed. The maximum gradient values are close to those in mammals. The plasma protein distribution between the intraand extravascular body fluids depends on the annual cycle period in the particular season (spawning and feeding completion) and the barrier properties of the capillary wall (peritoneum, brain, and white muscles). Among the osmotic reflection coefficient values characterizing the barrier properties of the capillary wall, those corresponding to its absolute impermeability were not found. This strategy can be explained by the fact that fish have to sustain the fluctuations in the intravascular fluid volume and its rapid recovery after hypovolemia, as well as by the features of protein metabolism in fish.

Macromolecular Distribution Volume in Human Ovarian and Endometrial Cancer

Elements of the extracellular matrix (ECM), notably collagen and glucosaminoglycans, will restrict part of the space available for soluble macromolecules because the molecules cannot occupy the same space. This phenomenon may influence macromolecular drug uptake. To study the influence of steric and charge effects of the ECM on the distribution volumes of macromolecules in human healthy and malignant gynecologic tissues we used as probes 15 abundant plasma proteins quantified by high-resolution mass spectrometry. The available distribution volume (VA) of albumin was increased in ovarian carcinoma compared with healthy ovarian tissue. Furthermore, VA of plasma proteins between 40 and 190 kDa decreased with size for endometrial carcinoma and healthy ovarian tissue, but was independent of molecular weight for the ovarian carcinomas. An effect of charge on distribution volume was only found in healthy ovaries, which had lower hydration and high collagen content, indicating that a condensed interstitium increases the influence of negative charges. A number of earlier suggested biomarker candidates were detected in increased amounts in malignant tissue, showing that interstitial fluid, even when unfractionated, can be a valuable source for tissue-specific proteins. We demonstrate that the distribution of abundant plasma proteins in the interstitium can be elucidated by mass spectrometry methods and depends markedly on hydration and ECM structure. Our data can be used in modeling of drug uptake, and give indications on ECM components to be targeted to increase the uptake of macromolecular substances.

Distribution volumes of macromolecules in human ovarian and endometrial cancers--effects of extracellular matrix structure.

Elements of the extracellular matrix (ECM), notably collagen and glucosaminoglycans, will restrict part of the space available for soluble macromolecules simply because the molecules cannot occupy the same space. This phenomenon may influence macromolecular drug uptake. To study the influence of steric and charge effects of the ECM on the distribution volumes of macromolecules in human healthy and malignant gynecologic tissues we used as probes 15 abundant plasma proteins quantified by high-resolution mass spectrometry. The available distribution volume (VA) of albumin was increased in ovarian carcinoma compared with healthy ovarian tissue. Furthermore, VA of plasma proteins between 40 and 190 kDa decreased with size for endometrial carcinoma and healthy ovarian tissue, but was independent of molecular weight for the ovarian carcinomas. An effect of charge on distribution volume was only found in healthy ovaries, which had lower hydration and high collagen content, indicating that a condensed interstitium increases the influence of negative charges. A number of earlier suggested biomarker candidates were detected in increased amounts in malignant tissue, e.g., stathmin and spindlin-1, showing that interstitial fluid, even when unfractionated, can be a valuable source for tissue-specific proteins. We demonstrate that the distribution of abundant plasma proteins in the interstitium can be elucidated by mass spectrometry methods and depends markedly on hydration and ECM structure. Our data can be used in modeling of drug uptake, and give indications on ECM components to be targeted to increase the uptake of macromolecular substances.

Age-related changes in rat dermal extracellular matrix composition affect the distribution of plasma proteins as a function of size and charge

Collagen and glycosaminoglycans (GAGs) constituting the ECM may limit the space available and thus exclude macromolecules from a fraction of the interstitial fluid (IF) phase. This exclusion phenomenon is of importance for transcapillary fluid and solute exchange. The purpose of the study was to examine the range of interstitial exclusion in rat skin by using probes within a span of molecular weights and electrical charge and also to test if a change in interstitial composition, occurring as a consequence of aging, affected exclusion. To this end, we used a novel approach, involving the exact determination of albumin concentration and mass in IF and tissue eluate by HPLC and thereafter, expressing the corresponding numbers relative to albumin for a set of probe proteins assessed by quantitative proteomics. Albumin was excluded from 55±4% (n=8) of the extracellular fluid phase. There was a highly significant, positive correlation between probe Stokes-Einstein (SE) radius and fractional excluded volume (VEF), described by VEF=0.078·SE radius+0.269 (P<0.001), and oppositely, a negative correlation between probe isoelectric point (pI) and exclusion for proteins with comparable size, VEF=-0.036·pI+0.719 (P=0.04). Aging resulted in a significant reduction in skin hydration and sulfated GAGs, a moderate increase in hyaluronan, and a corresponding, reduced VEF for albumin and the other macromolecular probes. Our findings suggest that the changes in the ECM in aged skin may result in delayed adjustments of fluid perturbations and reduced ability for salt storage.

Pharmacokinetics of Vilazodone in Patients with Mild or Moderate Renal Impairment

In patients with impaired renal function, the pharmacokinetics of a drug may be altered, resulting in decreased renal excretion or metabolism, and altered absorption, plasma protein binding or distribution. Vilazodone is a selective serotonin reuptake inhibitor and 5-HT1A receptor partial agonist approved for the treatment of major depressive disorder. Vilazodone is extensively hepatically metabolized with minimal renal excretion. The primary objective of this study was to assess the pharmacokinetics of a single 20-mg dose of vilazodone in subjects with mild or moderate renal impairment. This was a Phase 1, open-label, single-dose study of vilazodone in community-dwelling subjects with renal impairment and in healthy subjects to evaluate the pharmacokinetics of vilazodone in the presence of renal impairment. Thirty-two subjects were enrolled: eight subjects with mild (estimated glomerular filtration rate [est GFR] >50-80mL/min) renal impairment matched individually by age, sex and body mass index to eight control subjects with normal renal function (est GFR >80mL/min), and eight subjects with moderate (est GFR≥30-50mL/min) renal impairment matched with eight control subjects with normal renal function. Subjects received a single, 20-mg dose of vilazodone and pharmacokinetics, safety and plasma protein binding were assessed for 14days. The pharmacokinetic parameters calculated were maximum plasma concentration (Cmax), time to maximum plasma concentration, area under the plasma concentration-time curve (AUC) from time zero to 24h, AUC from time zero to the last measurable concentration, AUC from time zero to infinity estimated by linear trapezoidal rule and extrapolation, oral clearance, terminal elimination rate constant, elimination half-life (t½), free fraction in plasma, apparent free drug clearance, amount of vilazodone recovered in urine 0-96h following drug administration, percent of dose recovered in urine over 96h following drug administration, renal clearance and volume of distribution. Safety assessments were adverse events, clinical laboratory test results, 12-lead electrocardiograms and vital signs. Vilazodone pharmacokinetic parameters in renally impaired subjects were variable but not substantially different from healthy controls. Mean values for vilazodone Cmax and AUC were similar among groups. Mean t½ (35.7 and 34.8h mild and moderate vs. 37.0 and 34.8h matched controls), total drug clearance (19.9 and 25.1L/h vs. 26.4 and 26.9L/h), and mean vilazodone recovery in urine (1.21% and 0.58% vs. 0.95% and 0.81%) were similar for mild and moderate renally impaired subjects and matched controls with normal renal function. There were no apparent systematic trends in vilazodone pharmacokinetic parameters associated with decreasing renal function. Protein binding was variable (coefficient of variation, 29-65 %) but not substantially different among the three groups, and total drug clearance was not affected. Safety and tolerability of vilazodone were comparable in all groups of subjects. This study suggests that systemic exposure of vilazodone is not affected by mild or moderate renal impairment. No dose adjustments are recommended in patients with mild or moderate renal impairment.

Binding patterns of seminal plasma plasma proteins on bovine epididymal and ejaculated sperm membrane

The present study was designed to investigate the topographical distribution of seminal plasma (SP) proteins on epididymal and ejaculated bovine sperm. Using immunocytochemistry and confocal microscopy the binding patterns of bovine SP proteins BSP-A3, albumin, transferrin, prostaglandin D-synthase (PGDS) and nucleobindin in ejaculated and cauda epididymal sperm from adult bulls were evaluated. Experiments were performed using sperm from 5 males. Data showed a positive signal, only detected for anti-PGDS, in the acrosomal cap of epididymal and ejaculated sperm. In ejaculated sperm, a very weak signal for nucleobindin 2 in the midpiece and equatorial regions was detected, using the anti-rat nucleobindin. BSP-A3 was detected on all sperm regions studied, with a more evidenced signal in acrosome and midpiece. However, no binding was detected for albumin or transferrin in neither epididymal nor ejaculated sperm. In conclusion, PGDS, BSP-A3 and nucleobindin interact directly with bovine sperm, with specific topographic distribution. These findings may add to the knowledge of how these proteins modulate sperm functions, thus providing fundamental support for studies designed to evaluate how they influence sperm functions.

Reversible binding of the novel anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid to plasma proteins and its distribution into blood cells in various species.

The plasma protein binding and distribution in blood cells of the novel anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA) has been investigated in-vitro using filtration and an HPLC method to measure DMXAA. DMXAA (500 microM) was extensively bound in plasma from all species with an unbound fraction (fu) of 4.61+/-1.10 (mouse), 2.59+/-0.32 (rat), 2.02+/-0.48 (rabbit) and 2.07+/-0.23% (human). The binding was concentration dependent with DMXAA concentrations > or = 1,000 microM markedly increasing the fu in the plasma from all species. The estimated number of binding sites in plasma were 2.4+/-0.2 (mouse), 1.7+/-0.2 (rat), 0.8+/-0.1 (rabbit) and 2.1+/- 0.2 (human). The major binding protein in human plasma was albumin, with negligible binding to gamma-globulin and alpha1-acid glycoprotein. There was a significant linear relationship between the bound:free DMXAA concentration ratio (Cb/Cu) and albumin concentration in human serum albumin solution (r = 0.955; P < 0.05) and in healthy human plasma (r = 0.998; P< 0.05), but not in plasma from cancer patients (n = 5), nor across species. In cancer patients (n = 5) DMXAA had a significantly higher (P < 0.05) fu (4.60+/- 0.42%) compared with healthy human plasma (2.07+/-0.23%). In human plasma, the fu of DMXAA (500 microM) was significantly reduced by 500 microM diazepam (P < 0.05), but not by warfarin, phenylbutazone, salicylic acid, ibuprofen or clofibric acid at that concentration. DMXAA significantly reduced the binding of dansylsarcosine (a Site-II binder) to HSA, but significantly increased the binding of dansylamide (a Site-I binder). Within species, the blood:plasma concentration ratio (CBL/CP) of DMXAA was relatively constant (mouse, 0.581+/-0.005; rat, 0.667+/-0.025; rabbit, 0.637+/-0.019; human, 0.673+/-0.103) over the range 50-1000 microM, but increased significantly at DMXAA concentrations > 1000 microM in all species except the rabbit. These results indicate that significant alterations in DMXAA plasma binding and distribution into blood cells occur with increasing concentrations of DMXAA in all species, and also that significant interspecies differences exist. It would be more appropriate to compare plasma unbound concentrations when assessing DMXAA exposure in cancer patients or when extrapolating across species.

In vivo antioxidative activity of a quantified Pueraria lobata root extract

Aim of the study Oxidative stress has been associated with many pathological disorders such as atherosclerosis, diabetes and cancer. Supplementation with exogenous antioxidants, including phenolic compounds from plant sources, may help to restore the pro-oxidative/antioxidative balance. To take into account effects of absorption, metabolisation, plasma protein binding, distribution, and elimination, antioxidative research should not be limited to in vitro assays but be extended to in vivo models. Materials and methods In the present work a quantified 50% EtOH root extract of Pueraria lobata (Willd.) Ohwi (Fabaceae) was selected to determine its in vivo antioxidative activity in a diabetic rat model, where diabetes and the accompanying oxidative stress were induced by intraperitoneal administration of streptozotocin. This root extract was found to contain 10.42 ± 0.15% puerarin as the main constituent and smaller amounts of the related isoflavonoids 3′-hydroxypuerarin, 3′-methoxypuerarin, 6″-xylosylpuerarin, daidzin, genistin, daidzein and genistein, as determined by a validated HPLC method. This extract was administered orally at a daily dose of 500 mg/kg root extract, corresponding to 50 mg/kg puerarin, during 3 weeks. In addition the effect on the plasma concentration of some fat-soluble antioxidants (co-enzyme Q 9, α- and γ-tocopherol) was evaluated. Results and conclusions The level of malondialdehyde (MDA) in plasma, used as a marker of oxidative damage to lipids, was reduced to the same level as in healthy control animals, and as in the positive control group treated daily with 50 mg/kg α-tocopherol acetate. No obvious signs of toxicity were observed by administration of 10× the treatment dose.

Effect of colostrum immunoglobulin concentration on immunity in Majorera goat kids

The aim of the research was to evaluate the effects of immunoglobulin G (IgG) colostrum concentration on goat kid immune status when the total amount of IgG fed was constant. Majorera goat kids (n=56) were randomly assigned to 1 of 4 groups, and kids received 4g of IgG per kg of body weight of atomized colostrum at 4 different IgG concentrations: 20 (AC-20), 40 (AC-40), 60 (AC-60), and 80 (AC-80) mg/mL. Blood samples were obtained on d 0, 1, 2, 3, 4, and 5 postpartum. Immunoglobulin G, IgA, and IgM plasma concentrations, apparent efficiency of absorption of IgG, plasma chitotriosidase activity, plasma complement activity, and plasma proteinogram were measured. Plasma IgG and IgM concentrations were highest on d 1 in AC-80 animals, and IgA plasma concentration was lower in AC-20 than in AC-80. The apparent efficiency of absorption was higher in AC-80 (24.4%) than in the other treatment groups (by an average of 13.8%). Chitotriosidase plasma activity on d 5 (1,488 nmol/mL per hour) was higher than on d 0 and 1 (average of 1,183 nmol/mL per hour). There were no effects of colostrum IgG concentration on complement activity and plasma protein distribution, but γ-globulin and α-globulin were lower on d 0 than on d 1, 2, 3, 4, and 5. Increasing the immunoglobulin concentration in colostrum using atomized colostrum improves the immunoglobulin absorption at the same amount of immunoglobulin fed.