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PubMed1997

Cellular retention of liposome-delivered anionic compounds modulated by a probenecid-sensitive anion transporter.

PURPOSE: Drug carriers such as liposomes may enhance the intracellular delivery of therapeutic agents for infectious or neoplastic diseases. However, the mechanisms affecting cellular retention of liposome contents are understood poorly. We tested the hypothesis that retention of anionic compounds may be modulated by a nonspecific probenecid-sensitive anion transport mechanism, and that liposome composition may determine the impact of such transporters on drug retention by cells. METHODS: The fluorescent anionic dye hydroxy-pyrene-[1,3,6]-trisulfonate (HPTS) was transferred to the cytoplasm of cultured CV-1 or J774 cells by direct needle-microinjection or by ATP-induced permeabilization of the-plasma membrane, respectively, to investigate whether the cells have anion transport mechanisms capable of extruding HPTS from the cytoplasm. Cellular retention of dye was monitored in the presence and absence of the anion transport inhibitors probenecid or sulfinpyrazone. Liposomes containing HPTS were co-labeled with tetramethylrhodamine-labeled phosphatidylethanolamine (Rho-PE) as a marker of liposome membrane fate, and uptake was investigated using J774 cells. RESULTS: Needle-injected HPTS underwent both sequestration in early endocytic vesicles and rapid extrusion into the extracellular medium. Probenecid or sulfinpyrazone reduced the extrusion of HPTS. Thus HPTS is a substrate for a probenecid-sensitive anion transporter in J774 and CV1 cells. After delivery via fluid liposomes composed of phosphatidylglycerol:phosphatidylcholine:cholesterol (3:7:5 mole ratio) and co-labeled with Rho-PE, cell-associated HPTS declined more rapidly than did Rho-PE. Exposure of cells to 5 mM probenecid doubled the quantity of HPTS retained by cells, without changing the retention of the Rho-PE membrane marker. In contrast, the effect of probenecid was negligible when gel-phase liposomes of distearoylphosphatidylglycerol:cholesterol (10:5 mole ratio) were used. CONCLUSIONS: Probenecid-sensitive nonspecific anion transporters can mediate the extrusion of model anions delivered via liposomes. However, liposome composition modulates the amount of material subject to extrusion from cells, possibly by altering the endocytic compartment in which liposomes release their contents.

Animals↗
PubMed2003

Predictors of retention in dual-focus self-help groups.

Attendance at 12-step groups has been found useful in maintaining abstinence from substance use; many members disengage early, missing out on potential benefits. New 12-step based groups have emerged to address the recovery needs of the many substance users with psychiatric comorbidity. Little is known about factors associated with retention in 12-step, especially in this population. This study sought to identify predictors of retention over a one-year period among members of a dual-focus 12-Step fellowship (N = 276). Using multivariate analysis, the following baseline characteristics were associated with greater retention one year later: older age, more lifetime arrests, abstinence in the pre-baseline year, more psychiatric symptoms in the pre-baseline year, not taking psychiatric medication, being more troubled by substance abuse than by mental health, and greater level of self-efficacy for recovery; residing in supported housing and being enrolled in outpatient treatment at follow-up were also significantly associated with better retention. Clinical implications to enhance retention in specialized 12-step groups are discussed.

Adult↗
PubMed2001

Therapeutic alliance as a predictor of outcome and retention in the National Institute on Drug Abuse Collaborative Cocaine Treatment Study.

The authors examined the relation between therapeutic alliance, retention, and outcome for 308 cocaine-dependent outpatients participating in the National Institute on Drug Abuse Collaborative Cocaine Treatment Study. High levels of alliance were observed in supportive-expressive therapy (SE), cognitive therapy (CT), and individual drug counseling (IDC), and alliance levels increased slightly but significantly from Session 2 to Session 5 in all groups. In contrast to other studies, alliance was not a significant predictor of drug outcome. However, alliance did predict patient retention differentially across the 3 treatments. In SE and IDC, either higher levels of alliance were associated with increased retention or no relationship between alliance and retention was found, depending on the time alliance was measured. In CT, higher levels of alliance were associated with decreased retention.

Adult↗
PubMed1992

An endoplasmic reticulum retention signal in the CD3 epsilon chain of the T-cell receptor.

Isolated polypeptide chains of the T-cell antigen receptor complex are degraded or retained in the endoplasmic reticulum (ER). Assembly of the multisubunit complex allows the individual chains to escape retention in the ER and to be expressed on the cell surface. We engineered a series of deletions in the CD3 epsilon subunit of the human T-cell receptor in order to find the sequences responsible for its retention in the ER. Deletion of amino acids 171 to 180 in the cytosolic tail resulted in the cell-surface expression of the isolated chain. This sequence also promotes retention when it is appended to CD4, a plasma membrane protein. Mutagenesis of the 10-amino-acid CD3 epsilon sequence established that the tyrosine and serine residues are important for ER retention. This and other ER retention signals must be hidden when a complete T-cell receptor complex is assembled in order to allow its expression on the cell surface.

Amino Acid Sequence↗
PubMed2001

Retention characteristics of porous graphitic carbon in reversed-phase liquid chromatography with methanol-water mobile phases.

The solvation parameter model is used to study the retention mechanism of neutral organic compounds on porous graphitic carbon with methanol-water mobile phases containing from 0-100% (v/v) methanol. The dominant contribution to retention is the cavity formation-dispersion interaction term, composed of favorable interactions in the mobile phase (hydrophobic effect) and additional contributions from adsorption on the graphite surface. Electron lone pair and dipole-type interactions in the adsorbed state result in increased retention. Hydrogen-bonding interactions are more favorable in the mobile phase resulting in lower retention. The changes in the system constants of the solvation parameter model for cavity formation-dispersion interactions and hydrogen-bond interactions are linearly related to the volume fraction of water in the mobile phase. The system constants for electron lone pair interactions and dipole-type interactions are non-linear and go through a maximum and minimum value, respectively, at a specific mobile phase composition. The solvation parameter model poorly predicts the retention properties of angular molecules. This is probably due to the failure of the characteristic volume to correctly model the contact surface area for the interaction of angular molecules with the planar graphite surface. General factors affecting the quality of model fits for adsorbents are discussed.

Journal Article↗
PubMed1971

Testosterone action in the rat ventral prostate. The effects of diethylstilboestrol and cyproterone acetate on the metabolism of ( 3 H)testosterone and the retention of labelled metabolites by rat ventral prostate in vivo and in vitro.

Recent reports have indicated that the prior metabolism of testosterone by the secondary sexual tissues may be necessary for its androgenic effect. The effects of two anti-androgens, diethylstilboestrol and cyproterone acetate (17alpha-acetoxy-6-chloro-1,2alpha-methylenepregna-4,6-diene-3,20-dione) used in the chemotherapy of human prostatic carcinoma, have been examined on both the metabolism of testosterone and the retention of its metabolites by the rat ventral prostate gland. Cyproterone acetate was found to inhibit the retention of labelled metabolites of [(3)H]-testosterone by prostatic nuclei, both in vivo and in vitro. This inhibition appeared to be competitive. In contrast with its effect on nuclear retention of metabolites of testosterone, cyproterone acetate had no significant effect on the metabolism of [(3)H]testosterone by rat ventral prostate tissue. Diethylstilboestrol similarly had little effect on the metabolism of [(3)H]testosterone by prostatic tissue, although it did appear partially to inhibit its initial metabolism in all the incubation systems used. Diethylstilboestrol inhibited the nuclear retention of dihydrotestosterone when both [(3)H]testosterone and diethylstilboestrol were injected intraperitoneally in vivo, but had no effect on dihydrotestosterone retention when both testosterone and diethylstilboestrol were supplied directly to the prostate either in vivo or in vitro. It was concluded that if diethylstilboestrol has an anti-androgenic effect at the level of the target organ as distinct from its effect on androgen production by the testes, then it is probably due to a mechanism differing from that of cyproterone acetate.

Androgen Antagonists↗
PubMed2003

The investigation of occlusal contacts during the retention period.

Removable retention appliances were applied to 20 treated orthodontic patients, and their occlusal contact points were determined from occlusal registrations taken at the beginning and the end of retention. Furthermore, to determine the results at the end of retention, the occlusion of 20 treated patients was compared with a control group of another 20 subjects who had an ideal occlusion. During the retention phase the number of contacts in centric occlusion increased significantly. No significant difference was observed with regard to the location of contacts. Whereas the number of ideally located contacts was similar to that in the control group, differences were observed in actual and canine contacts between the groups. At the end of retention, the balancing side contacts in lateral movements and the posterior contacts in protrusive movements were generally determined as near contacts and showed a similarity to the control group. In order to maintain the occlusal stability that is needed for the success of orthodontic treatments, ideal occlusal contacts and localization of contacts in centric and eccentric occlusion should be considered.

Adolescent↗
PubMed2007

The critical shortage of speech-language pathologists in the public school setting: features of the work environment that affect recruitment and retention.

PURPOSE: The primary focus of this study was to elicit the perspectives of speech-language pathologists (SLPs) regarding features of the work environment that contribute to and/or hinder recruitment and retention in the public school setting. METHOD: A questionnaire was distributed to SLPs employed in 10 school districts in Central Florida representing small, medium, and large school districts. The primary goal of the questionnaire was to elicit the perspectives of school-based SLPs regarding (a) factors in the work environment that contribute to retention, (b) factors in the work environment that hinder retention, and (c) issues that may contribute to the recruitment and retention of SLPs in the school setting. A total of 382 questionnaires was obtained, yielding a 64.5% response rate. RESULTS: The participants ranked working with children, school schedule, and educational setting as primary reasons for their satisfaction with working in the public school setting. The participants ranked workload, role ambiguity, salary, and caseload as primary reasons for their dissatisfaction with working in the public school setting. CONCLUSION: Themes emerged from the data that provide insight into several factors that serve as powerful influences in understanding issues of recruitment and retention of SLPs in the public school setting.

Humans↗
PubMed2003

Improving endothelial cell retention for single stage seeding of prosthetic grafts: use of polymer sequences of arginine-glycine-aspartate.

OBJECTIVE: single stage seeding within the timeframe of a typical vascular operation has not been successful. One reason for this is poor cell adherence to the graft lumen once exposed to pulsatile blood flow. In this study we have carried out investigations with the use of two different fibronectin-based peptides, fibronectin-like engineered protein polymer (FEPP) which contains multiple copies of arginine-glycine-aspartate (RGD) and fibronectin adhesion promoting peptide (FAPP) to improve cell adherence. MATERIALS AND METHODS: FAPP and FEPP were coated onto native polyurethane and heparinised polyurethane grafts. The grafts were then seeded for either 1 or 2h with human umbilical vein endothelial cells (HUVEC). After the incubation period the cells were washed off and cell retention was calculated. Cell metabolism was measured using Alamar Blue, and confirmed with scanning electron microscopy (SEM). RESULTS: heparinised grafts coated with FEPP showed the best cell retention after both 1 and 2h seeding (80+/-4% vs 81+/-3%). This graft had no significant difference in cell retention after both times whilst all the other grafts had better cell retention after a 2h seeding. The Alamar blue and SEM results confirmed cell viability and function for all graft types. CONCLUSION: heparinised graft coated with FEPP allows significant cell retention after only 1h of seeding and shows promise for single stage seeding.

Blood Vessel Prosthesis↗
PubMed1998

Rapid palatal expansion. Part 3: strains developed during active and retention phases.

Patterns of strain accumulation/dissipation during the active and retention phases of rapid palatal expansion treatment were studied in a preliminary animal model (5 cats) followed by clinical study (14 patients). Two uni-axial strain gauges were bonded to the arms of a hyrax screw. The strain gauges were wired intraorally to a common male connector and protected against salivary assault. For strain registration, the connector was hooked to a switch and balance instrument and a digital strain indicator. The screw was activated (4 x 1/4 turn) every 3 to 4 days and strain and interarch distances were measured during rapid palatal expansion active and retention phases. Interarch distances were also measured during the relapse phase. Both studies share the same results. Strains and expansion increased progressively during active and retention phases of rapid palatal expansion. No difference between anterior and posterior strain was found. An immediate dental strain response that occurred during screw activations (intrasession strain) was related to tooth compression in the PDL. During the 3 to 4 day pause, a delayed skeletal strain response (intersession strain) developed that was inverted or continuous to the preceding intrasession strain. Complete dissipation of residual strains was limited to the first 1 to 3 active phase sessions. Strain accretion resulted essentially from intersession strain build-up. Strain level was preserved during the retention phase, apparently due to relapse strains. Relapse strains could not be measured but are inferred from the predominant interarch rebound measured during the relapse phase. Clinically, an extension of the intersession intervals and the retention phase are recommended.

Adolescent↗
PubMed2002

Low-dose desmopressin infusion: renal action in healthy women in moderate salt retention and depletion, and interactions with prostanoids.

Studies were carried out to evaluate the influence of variations in sodium balance on the renal response to low-dose infusion of 1-desamino-8- D -arginine vasopressin (dDAVP), and the functional interaction between dDAVP and renal prostanoids. The studies were performed on healthy women in conditions of extracellular fluid volume expansion (SR group, n =9) and depletion (SD2 group, n=6), respectively. The study protocol included hypotonic polyuria (induced by oral water load) and subsequent antidiuresis (induced by low-dose infusion of dDAVP). Three 60-min clearance (cl.) periods were performed during polyuria (cl. P), early (cl. A1) and late (cl. A2) antidiuresis. The urinary concentrations of prostaglandin (PG) E(2) and the stable metabolites of PGI(2) and thromboxane (Tx) A(2), 6-keto-PGF(1alpha) (6KPGF) and TxB(2), were estimated. Paired renal functional explorations were performed in salt retention and salt depletion both in absence and presence of indomethacin (SR.I and SD2.I groups). In both paired and unpaired studies, the early and late effects of dDAVP on the functional excretory variables and the excretion of prostanoids were assessed as percentage variations, (A1-P)% P and (A2-A1)% A1. (I) dDAVP in salt retention and depletion. During early infusion dDAVP produced in both conditions a significant reduction in urinary flow rate, creatinine cl., absolute and fractional excretions of sodium, chloride and potassium; during late infusion dDAVP was effective in inducing a further significant reduction in urinary flow rate. In salt retention compared to depletion the early reductions in sodium and chloride (absolute and fractional) excretions were significantly lower. (II) Indomethacin pretreatment. During early infusion the dDAVP-induced reductions in the urinary flow rate and 6KPGF excretion were enhanced in both conditions. In salt depletion the dDAVP effects in reduction of creatinine cl. and urinary electrolyte excretions were also enhanced. During late infusion the antidiuretic effect of dDAVP was suppressed in salt retention, while in salt depletion creatinine cl., the urinary excretions of electrolytes and both 6KPGF and TxB(2) showed increases significantly different from the dDAVP effects in the absence of indomethacin. In conclusion, (a) the salt-retaining effect of dDAVP was less effective in salt retention compared to depletion. (b) Indomethacin pretreatment affected the renal action of dDAVP in a time-dependent pattern. The early effects in both conditions were consistent with an inhibited synthesis of modulator PGs. On the contrary, the late effects were consistent with the occurrence, at least in salt depletion, of an escape from dDAVP renal action. This escape phenomenon probably depended on a partial regression of the pharmacological inhibition of the modulating PGs.

Adult↗
PubMed1986

[Comparison of the renal ultrasound-furosemide test with the isotope nephrogram. Possible clue to the retention phenomenon?].

In 23 patients with essential hypertension renographic and sonographic examination of the kidneys was compared. No correlation could be observed for functional parameters and urinary excretion between the two different methods. Correlations have only been detected between renographic retentions and furosemide-conditioned swelling of the kidneys in sonography: patients with severe retention showed a statistically significant stronger response to furosemide. The increased retention in the renal pelvis could be responsible for alterations in sonographic findings. This study provides new data on the mechanisms of retention, whereby retention in the renal pelvis in some patients seems to be most probably caused by nephrosclerosis.

Adult↗
PubMed2000

In vivo retention of endothelial cells adenovirally transduced with tissue-type plasminogen activator and seeded onto expanded polytetrafluoroethylene.

PURPOSE: Seeding a prosthetic graft with genetically engineered vascular endothelial cells (ECs) has the potential to enhance the graft's antithrombotic properties. However, it has been reported that ECs transduced with tissue-type plasminogen activator (tPA) have very low levels of retention on grafts, probably because of increased proteolytic activity. We examined the retention of human tPA (htPA)-transduced ECs after the cells were seeded onto expanded polytetrafluoroethylene (ePTFE) grafts and implanted into dogs. We also examined the function of secreted htPA in this model. METHODS AND RESULTS: Canine jugular venous ECs were transduced with adenoviral vectors encoding htPA (Adex1CAhtPA) and beta-galactosidase (Adex1CALacZ). There was a positive relationship between the percentage of X-gal ECs staining and the multiplicity of infection (MOI) of Adex1CALacZ. The level of htPA production in vitro increased with the increasing MOI of Adex1CAhtPA, but decreased gradually 4 days after infection. ECs coinfected with Adex1CAhtPA and Adex1CALacZ (htPAEC) or ECs infected with Adex1CALacZ alone (LacZEC) were seeded onto ePTFE grafts at densities equivalent to confluence to visualize retained ECs in an in vivo flow study. The grafts were implanted into canine carotid arteries and harvested after 5 hours of exposure to blood flow. The harvested grafts showed patchy defects in ECs, most of which were covered with mural thrombi. There was no significant difference in retention between htPAEC (29.3% +/- 8.7%) and LacZEC (19.5% +/- 3. 6%). There was a significant negative correlation between the in vivo EC retention on the grafts and the in vitro cellular passage level of ECs (P =.041; r = -.40). htPAEC produced 210.3 +/- 22.2 ng htPA antigen/10(6) cells per 6 hours in vitro and continued to secrete htPA on the harvested graft. CONCLUSIONS: We demonstrated that a large amount of functional htPA was produced by adenovirally modified canine ECs. The results of the in vivo study may suggest that overexpression of tPA has little effect on the short-term retention of early passage ECs seeded onto ePTFE grafts.

Adenoviridae↗
PubMed2001

A critical review of rural medical workforce retention in Australia.

The problem of how best to recruit and retain doctors in rural and remote communities has led governments to adopt a range of medical workforce incentives, including retention grants. A comprehensive literature survey suggests that medical workforce retention has been poorly distinguished from other supply issues such as recruitment, and that its determinants and the process leading to retention are poorly understood. Such a knowledge gap is likely to limit the effectiveness of retention incentives. This article reports the results of this literature review, and advances a conceptual framework as the basis for ongoing research and evaluating how best to deliver effective retention interventions.

Australia↗
PubMed2003

A high-molecular-weight complex of membrane proteins BAP29/BAP31 is involved in the retention of membrane-bound IgD in the endoplasmic reticulum.

B cell antigen receptors (BCRs) are multimeric transmembrane protein complexes comprising membrane-bound immunoglobulins (mIgs) and Ig-alpha/Ig-beta heterodimers. In most cases, transport of mIgs from the endoplasmic reticulum (ER) to the cell surface requires assembly with the Ig-alpha/Ig-beta subunits. In addition to Ig-alpha/Ig-beta, mIg molecules also bind two ER-resident membrane proteins, BAP29 and BAP31, and the chaperone heavy chain binding protein (BiP). In this article, we show that neither Ig-alpha/Ig-beta nor BAP29/BAP31 nor BiP bind simultaneously to the same mIgD molecule. Blue native PAGE revealed that only a minor fraction of intracellular mIgD is associated with high-molecular-weight BAP29/BAP31 complexes. BAP-binding to mIgs was found to correlate with ER retention of chimeric mIgD molecules. On high-level expression in Drosophila melanogaster S2 cells, mIgD molecules were detected on the cell surface in the absence of Ig-alpha/Ig-beta. This aberrant transport was prevented by coexpression of BAP29 and BAP31. Thus, BAP complexes contribute to ER retention of mIg complexes that are not bound to Ig-alpha/Ig-beta. Furthermore, the mechanism of ER retention of both BAP31 and mIgD is not through retrieval from a post-ER compartment, but true ER retention. In conclusion, BAP29 and BAP31 might be the long sought after retention proteins and/or chaperones that act on transmembrane regions of various proteins.

Animals↗
PubMed1998

Hepatitis C virus glycoprotein complex localization in the endoplasmic reticulum involves a determinant for retention and not retrieval.

The hepatitis C virus (HCV) genome encodes two envelope glycoproteins (E1 and E2). These glycoproteins interact to form a noncovalent heterodimeric complex which in the cell accumulates in endoplasmic reticulum (ER)-like structures. The transmembrane domain of E2, at least, is involved in HCV glycoprotein complex localization in this compartment. In principle, ER localization of a protein can be the consequence of actual retention in this organelle or of retrieval from the Golgi. To determine which of these two mechanisms is responsible for HCV glycoprotein complex accumulation in the ER, the precise localization of these proteins was studied by immunofluorescence, and the processing of their glycans was analyzed. Immunolocalization of HCV glycoproteins after nocodazole treatment suggested an ER retention. In addition, HCV glycoprotein glycans were not modified by Golgi enzymes, indicating that the ER localization of these proteins is not because of their retrieval from the cis Golgi. Retention of HCV glycoprotein complexes in the ER without retrieval suggests that this compartment plays an important role for the acquisition of the envelope of HCV particles. A true retention in the ER was also observed for E2 expressed in the absence of E1 or for a chimeric protein containing the ectodomain of CD4 in fusion with the transmembrane domain of E2. These data indicate that, in HCV glycoprotein complex, the transmembrane domain of E2, at least, is responsible for true retention in the ER, without recycling through the Golgi.

Animals↗
PubMed2004

An improved model for prediction of retention times of tryptic peptides in ion pair reversed-phase HPLC: its application to protein peptide mapping by off-line HPLC-MALDI MS.

The proposed model is based on the measurement of the retention times of 346 tryptic peptides in the 560- to 4,000-Da mass range, derived from a mixture of 17 protein digests. These peptides were measured in HPLC-MALDI MS runs, with peptide identities confirmed by MS/MS. The model relies on summation of the retention coefficients of the individual amino acids, as in previous approaches, but additional terms are introduced that depend on the retention coefficients for amino acids at the N-terminal of the peptide. In the 17-protein mixture, optimization of two sets of coefficients, along with additional compensation for peptide length and hydrophobicity, yielded a linear dependence of retention time on hydrophobicity, with an R2 value about 0.94. The predictive capability of the model was used to distinguish peptides with close m/z values and for detailed peptide mapping of selected proteins. Its applicability was tested on columns of different sizes, from nano- to narrow-bore, and for direct sample injection, or injection via a pre-column. It can be used for accurate prediction of retention times for tryptic peptides on reversed-phase (300-A pore size) columns of different sizes with a linear water-ACN gradient and with TFA as the ion-pairing modifier.

Amino Acid Sequence↗
PubMed1992

Involvement of food intake in the decreased energy retention associated with single deficiencies of lysine and sulphur-containing amino acids in growing chicks.

1. Growth and energy utilisation were determined in growing chicks fed ad libitum on diets deficient either in lysine (5.95 g/kg) or in sulphur-containing amino acids (SAA, 3.5 g/kg). Food intake, body weight gain, energy retained as protein and as fat, and total energy retention were significantly (P less than 0.05) reduced by single deficiencies of either lysine or SAA. 2. Another two experiments were conducted to determine if the decreased total energy retentions in chicks fed on diets deficient in lysine (experiment 3) or SAA (experiment 4) were associated with reduced food intake, by using tube-feeding to control the amount and pattern of food consumption. Chicks fed on diets deficient in lysine or SAA retained less energy as protein and more energy as fat than the control chicks. Neither total energy retention nor heat increment was affected by these deficiencies. Total energy retention was proportional to metabolisable energy (ME) intake alone. 3. It is concluded that the decreased total energy retentions caused by single deficiencies of lysine and SAA were associated with decreased food intake.

Amino Acids, Sulfur↗