Pharmacokinetic study of saquinavir hard gel caps/ritonavir in HIV-1-infected patients: 1600/100 mg once-daily compared with 2000/100 mg once-daily and 1000/100 mg twice-daily.

OBJECTIVES: A pharmacokinetic comparison of three dosing regimens of saquinavir/ritonavir was carried out: 1600/100 mg once-daily with 1000/100 mg twice-daily, and 1600/100 mg once-daily with 2000/100 mg once-daily. METHODS: Twenty patients on saquinavir hard gel caps/ritonavir 1600/100 mg once-daily in combination with two nucleoside reverse transcriptase inhibitors for at least 4 weeks were enrolled and randomized to either saquinavir hard gel caps/ritonavir 1000/100 mg twice-daily or 2000/100 mg once-daily. Two pharmacokinetic curves were plotted, at baseline (day 0) and 7 days after the switch. Plasma concentrations were measured at 0, 2, 4, 6, 8, 10, 12 (and 24 for once-daily dosing) hours after drug intake by validated high-performance liquid chromatographic assay (HPLC). The area under the plasma concentration-time curve (AUC0-24 or AUC0-12), maximum and minimum concentration (Cmax and Cmin) and elimination half-life were calculated using a non-compartmental model. RESULTS: Compared with saquinavir/ritonavir 1600/100 mg once-daily dosing, the saquinavir AUC and Cmin improved significantly when dosed as 1000/100 mg twice-daily (53% and 299%, respectively), and as 2000/100 mg once-daily (71% and 65%, respectively). Low Cmin in three subjects at baseline was corrected after switch to the other dosages. Saquinavir/ritonavir 2000/100 mg once-daily was also associated with a significant increase in saquinavir Cmax (52%) compared with saquinavir/ritonavir 1600/100 mg once-daily. CONCLUSIONS: Saquinavir/ritonavir when dosed as 2000/100 mg once-daily or 1000/100 mg twice-daily achieves higher saquinavir plasma levels compared with saquinavir/ritonavir 1600/100 mg once-daily. Taking the convenience of once-daily dosing into consideration, dosage of 2000/100 mg once-daily may be preferred.

New transport peptides broaden the horizon of applications for peptidic pharmaceuticals.

Protein transduction domains (PTDs) have proven to be an invaluable tool to transduce a wide variety of cargo's including peptides across the plasma membrane and into intact tissue. The PTDs are able to deliver biologically active molecules both in vitro and in vivo. This study describes many new polybasic PTDs of which some are just as potent as the PTDs derived from extracellular RNAses or other published PTDs. Large differences in potency became apparent when the PTDs are coupled to particular cargoes. Therefore, the unique characteristic of a PTD may only become apparent when it is selected for a particular application. Rules for optimization of PTDs for particular applications are now emerging and open the way for a new generation of drug delivery agents. Because fixation artifacts and irreversible membrane binding may cause misinterpretation of the amount of internalization of polybasic peptides, we have developed an enzyme transduction assay based on the intracellular loading of a cell permeable substrate. In this assay, a fluorescent signal is generated by internalized enzyme in intact cells and not by membrane-bound or extracellular enzyme.

Design and synthesis of a multivalent homing device for targeting to murine CD22.

CD22 is a cell-surface glycoprotein uniquely located on mature B-cells and B-cell derived tumour cells. Current evidence suggests that binding of endogenous ligands to CD22 leads to modulation of B-cell activation by antigen. Incidentally, however, B-cell activation may derail. and lead to an undesired immune response, for example in cases of allergy, rheumatoid arthritis and Crohn's disease. In this situation, synthetic high-affinity ligands for CD22 may be of therapeutic value as inhibitors of B-cell activation. Recent studies have revealed that natural ligands for CD22 contain the trisaccharide NeuAc alpha-2,6-Lac as the basic binding motif. In addition, it has been demonstrated that binding to CD22 is strongly enhanced by multivalent presentation of the basic binding motif (cluster effect). In this paper. the stepwise development of a novel multivalent high-affinity ligand for CD22 is described. In the first stage, a series of monovalent NeuAc alpha-2,6-Glc(Y)X type binding motifs was prepared, and their affinity for murine CD22 was monitored, to obtain more insight into the effect of separate structure elements on ligand recognition. In the second stage, we prepared a trivalent cluster, based on the monovalent motif that displayed the highest affinity for CD22, NeuAc alpha-2,6-GlcNBzNO2OMe (7). This cluster, TRIS(NeuAc alpha-2,6-GlcNBzNO2)3 (52), displayed a more than 58-fold higher affinity for CD22 than the reference structure NeuAc alpha-2,6-LacOMe (10). To our knowledge, the cluster 52 is one of the most potent antagonists for CD22 yet synthesised.

A structure-function study of ligand recognition by CD22beta.

B-cell-specific CD22 is a member of a group of cell adhesion molecules within the immunoglobulin superfamily that display binding to glycans with terminal sialic acid residues. Binding of endogenous ligands to CD22 triggers B-cell activation and proliferation. It is therefore conceivable that high affinity ligands for CD22 may be of value as inhibitors of B-cell activation in allergy and chronic inflammation. In this study, we aimed to delineate the structural requirements for ligand binding to CD22. A library of 20 mono-, di-, and trisaccharide analogs of the basic binding motif Neu5Ac(alpha2,6)Lac was synthesized and screened for affinity for CD22beta. In general, CD22 ligand recognition appeared to be rather tolerant with respect to structural modifications of the anomeric sugar on a mono-, di-, and trisaccharide level, although affinity was increased by the presence of a nitro aromatic group at C-2. The most potent monovalent ligand, Neu5Ac-4-nitrobenzoyl-Glc, was selected to generate multivalent ligands based on either a glutamate or Tris cluster core. All multivalent ligands displayed at least a 10-fold increased affinity for CD22 compared with the corresponding monovalent glycoside. Interestingly, a maximal gain in affinity was already obtained for bivalent ligands, regardless of the terminal glycoside. A trivalent Tris-based cluster of Neu5Ac-4-nitrobenzoyl-Glc displayed a 300-fold higher affinity compared with the basic binding motif, which makes it, to our knowledge, the most potent antagonist for CD22 yet synthesized. As our in vitro fluorescence-activated cell sorting studies demonstrated efficient cellular uptake of a CD22 substrate, the most potent ligand in this study may hold promise as a homing device for immunomodulatory compounds and cytostatics.

Deep vein thrombosis: clinical nursing management.

The occurrence of a deep vein thrombosis is potentially life threatening and rapid assessment and treatment are essential to prevent development of a pulmonary embolism. Prophylaxis and risk assessment are important aspects of DVT management.

Calculating patients' risk of deep vein thrombosis.

Deep vein thrombosis (DVT) constitutes a serious threat to patients' general recovery. DVT in postoperative and bedridden patients is usually preventable and a thromboprophylaxis protocol based on risk assessment categories is strongly recommended. The Autar DVT risk assessment scale was developed to separate risk into no risk, low, moderate and high risk categories. Founded on Virchow's triad in the genesis of DVT, the scale is composed of seven categories of risk factors. When the scale was tested on a trauma/orthopaedic unit a cut-off score of 16 yielded 100% sensitivity, 81% specificity and a correlation coefficient of 0.98. The DVT scale is designed to allow application in diverse clinical specialties. It is recommended that nurses using the Autar DVT scale should evaluate for themselves the best cut-off score to achieve maximum predictive accuracy.

Renal clearance of the thyrotropin-releasing hormone-like peptide pyroglutamyl-glutamyl-prolineamide in humans.

TRH-like peptides have been identified that differ from TRH (pGlu-His-ProNH2) in the middle amino acid. We have estimated TRH-like immunoreactivity (TRH-LI) in human serum and urine by RIA with TRH-specific antiserum 8880 or with antiserum 4319, which binds most peptides with the structure pGlu-X-ProNH2. TRH was undetectable in serum (< 25 pg/mL), but TRH-LI was detected with antiserum 4319 in serum of 27 normal subjects, 21 control patients, and 12 patients with carcinoid tumors (range 17-45, 5-79, and 18-16,600 pg/mL, respectively). Because serum was kept for at least 2 h at room temperature, which causes degradation of TRH, pGlu-Phe-ProNH2, and pGlu-Tyr-ProNH2, serum TRH-LI is not caused by these peptides. On high-performance liquid chromatography, serum TRH-LI coeluted with pGlu-Glu-ProNH2 (< EEP-NH2), a peptide produced in, among others, the prostate. Urine of normals and control patients also contained TRH-LI (range 1.14-4.97 and 0.24-5.51 ng/mL, respectively), with similar levels in males and females. TRH represented only 2% of urinary TRH-LI, and anion-exchange chromatography and high-performance liquid chromatography revealed that most TRH-LI in urine was < EEP-NH2. In patients with carcinoid tumors, increased urinary TRH-LI levels were noted (range 1.35-962.4 ng/mL). Urinary TRH-LI correlated positively with urinary creatinine, and the urinary clearance rate of TRH-LI was similar to the glomerular filtration rate. In addition, serum TRH-LI was increased in 17 hemodialysis patients (43-373 pg/mL). This suggests that serum < EEP-NH2 is cleared by glomerular filtration with little tubular resorption. The possible role of the prostate as a source of urinary TRH-LI was evaluated in 11 men with prostate cancer, showing a 25% decrease in urinary TRH-LI excretion after prostatectomy (0.19 +/- 0.02 vs. 0.15 +/- 0.01 ng/mumol creatinine, mean +/- SEM). However, TRH-LI was similar in spontaneously voided urine and in urine obtained through a nephrostomy cannula from 16 patients with unilateral urinary tract obstruction (0.15 +/- 0.01 vs. 0.14 +/- 0.01 ng/mumol creatinine). These data indicate that: 1) TRH-LI in human serum represents largely < EEP-NH2, which is cleared by renal excretion; 2) part of urinary < EEP-NH2 is derived from prostatic secretion into the blood and not directly into urine; and 3) urinary < EEP-NH2 can be used as marker for carcinoid tumors.

The scope of professional practice in specialist practice.

The Scope of Professional Practice (UKCC, 1994a) now offers nurses, midwives and health visitors new opportunities to develop their clinical role. Role expansion is at the centre of debate in the health service. In the spirit of Scope, trusts and regional health authorities have been experimenting with new roles that combine medical and nursing practice in order to promote a holistic approach to health care. Education, experience and the appropriate clinical competency are the foundations for role expansion and the development of professional practice. However, some of the new roles and innovative practices have also been targeted at reducing junior doctors' hours (Read and Graves, 1994). New roles must be evaluated before they are allowed to develop and there is an urgent need for national evaluation of the initiatives underpinning Scope. This article examines the changing role of practitioners in specialist practice. The ethical, professional and legal issues surrounding these changes will also be considered.

Nursing assessment of clients at risk of deep vein thrombosis (DVT): the Autar DVT scale.

Deep vein thrombosis (DVT) poses a threat to hospitalized clients' recovery. It is preventable and the cost of treating this problem is considerably more than that of preventative practices. Accurate DVT risk assessment facilitates the application of the most appropriate venous thromboprophylaxis. Founded on Virchow's triad of risk factors in the genesis of deep vein thrombosis, the Autar DVT scale was developed as a predictive index. The DVT scale is composed of the following seven risk categories: increasing age, build and body mass index (BMI), immobility, special DVT risk, trauma, surgery and high risk disease. The DVT scale was tested on two trauma wards and the study was essentially a data generating exercise. Clinical data were gathered on 21 clients to validate the reliability, sensitivity and specificity of the DVT scale. Pearson moment correlation coefficient (r) and total percentage agreement (T%) measurement yielded a value of r at 0.98 and a T% ranging between 70% and 87% respectively for both reliability studies. Predictive validity of the scale calculated from a threshold score of 16 achieved 100% sensitivity and 81% specificity. The Autar DVT scale has produced some interesting results and holds considerable promise as a predictive index. However, as this was a small study further testing in diverse clinical areas of a large client population is recommended.

Role of the nurse teacher in advanced nursing practice.

Education and experience are the foundations on which practitioners should base their clinical judgment and broaden the scope of their professional practice. PREP has secured a professional market for nurse education, the viability of which depends on closer collaboration between nursing education and nursing practice. Nurse teachers who are committed to practice are in a strong position to assist the development of practitioners, and work towards research-based practice. This article examines the role of the nurse teacher in the domain of advanced nursing practice.

Biphasic model of the trabecular meshwork in the eye.

A biphasic continuum model for the flow of intraocular fluid, the aqueous humour, in the trabecular meshwork is proposed in the paper. The model considers the meshwork as a circular ring with uniform thickness of homogeneous, isotropic, viscoelastic material swollen with continuously percolating aqueous humour. The model further assumes the permeability of the meshwork as a function of dilation in its solid phase. The study considers steady and quasisteady states. Approximate solutions to the problem are obtained by Picard's type iterative procedure and computational results for the dimensionless solid displacement, permeability and pressure profiles are presented. It is concluded that intraocular pressure has a permeability-decreasing effect, whereas the elasticity of the meshwork shows a permeability-increasing effect. Thus, rising intraocular pressure develops a further rise in itself, whereas the elasticity tends to diminish the intraocular pressure through facilitating aqueous outflow.

Flow of aqueous humor in the canal of Schlemm.

A mathematical model is presented for the flow of aqueous humor in Schlemm's canal in the eye. The model introduces a canal segment between two collector channels as a rectangular channel with porous upper wall. Two cases have been considered in the model: (I) the inner porous wall of the canal is rigid; (II) the inner wall is collapsible. Analytical solution of the governing equation in case I is straightforward, whereas the nonlinear equation in case II is solved by an iterative procedure. Aqueous fluid pressure and flow profiles in the proposed model are drawn, and the effects of important parameters on these profiles are brought out and discussed. It is concluded that for case I, resistance to aqueous flow is influenced by the filtration constant of the trabecular and endothelial meshwork and that narrowing of the canal reduces outflow. In case II, an increase in intraocular pressure (IOP) or compliance coefficient of the canal inner wall increases the collapse of the canal, which offers increased resistance to flow resulting in the decreased flow whereas increasing filtration constant facilitates aqueous outflow. These theoretical results suggest that increased IOP or decreased rigidity of the inner wall may contribute to the development of increased resistance as observed in some cases of glaucoma and that increasing values of filtration constant may contribute to the facility of outflow increase.