Continuous dosing of a novel contraceptive vaginal ring releasing Nestorone® and estradiol: pharmacokinetics from a dose-finding study.

BACKGROUND: As part of a program to develop a novel estradiol-releasing contraceptive vaginal ring (CVR), we evaluated the pharmacokinetic (PK) profile of CVRs releasing segesterone acetate (Nestorone® (NES)) combined with one of three different estradiol (E2) doses. STUDY DESIGN: A prospective, double-blind, randomized, multi-centered study to evaluate a 90-day CVR releasing NES [200mcg/day] plus E2, either 10mcg/day, 20mcg/day, or 40mcg/day in healthy reproductive-age women with regular cycles. Participants provided blood samples twice weekly for NES and E2 levels during the first 60 days (ring 1) and the last 30 days (ring 2) of use. A subset underwent formal PK assessments at ring initiation, ring exchange (limited PK), and study completion. RESULTS: The main study enrolled 197 women; 22 participated in the PK substudy. Baseline characteristics between the main and PK participants were comparable, with an average BMI of 25.8 kg/m2 (SD 4.3). In the PK substudy, all three rings showed similar NES PK: mean area under the curve (AUC(0-72)) 34,181 pg*day/mL; concentration maximum (Cmax) 918 pg/mL; time to maximum concentration (Tmax) 3.5 h. For E2, the Cmax occurred at 2 h, and was significantly higher with the 20 mcg/day ring (mean 390 pg/mL); 10 mcg/day, 189 pg/mL, p=.003; 40 mcg/day, 189 pg/mL, p<.001), and declined rapidly to≤50 pg/mL for all doses by 24 h. For all subjects, the median E2 levels remained under 35 pg/mL during treatment. CONCLUSION: PK parameters of NES were not affected when paired with different doses of E2, but E2 levels from all three doses were lower than anticipated and no dose response was observed. IMPLICATIONS: While these novel estradiol-releasing combination contraceptive vaginal rings provided sustained release of contraceptive levels of Nestorone over 90 days, the E2 levels achieved were not consistent with bone protection, and a dose-response was not observed.

Surface hydrophilisation and antibacterial functionalisation for microporous polypropylene membranes.

The surface properties of polymer membranes are crucial to their separation performances. For the microporous polypropylene membranes, the high hydrophobicity and lack of functionality easily cause protein adsorption and subsequent microorganism attachment and biofilm formation, i.e. biofouling. Thus, their applications in water treatment, bioseparation and biomedical fields are largely limited. Surface hydrophilisation and antibacterial functionalisation are, therefore, reasonably necessary. This review provides a concise summarisation of related studies according to the surface modification strategies. Especially, the interfacial crosslinking approach developed in our previous studies is presented in detail.

Kinetics of KB and HEp-2 cell responses to an invasive, cytolethal distending toxin-producing strain of Actinobacillus actinomycetemcomitans.

The periodontal pathogen Actinobacillus actinomycetemcomitans produces cytolethal distending toxin (CDT), a complex multicomponent toxin that arrests the growth of many types of eukaryotic cell. The kinetics of the effects of CDT-containing extracts, from an invasive strain of this bacterium, were examined on epithelial-like cells routinely used in invasion studies. Both KB and HEp-2 cells were exquisitely sensitive to the effects of the CDT with TD50 of 30 and 300 pg of total bacterial protein, respectively. Initial cell morphology changes were relatively rapid, occurring within the first 13 h of exposure. CDT-treated KB cells increased in size to 4-5 times the size of untreated controls. Cytotoxicity was irreversible when attached cells were incubated, for a minimum of 120 min, with nanogram quantities of CDT-containing extract. As cultures aged, the cells became more resistant to the effects of the CDT-containing extracts. These findings have important implications for understanding the ability of A. actinomycetemcomitans to invade and multiply in epithelial cells.

Endometrial brush biopsy for the diagnosis of endometrial cancer.

OBJECTIVE: To evaluate a new technique for processing endometrial cytology for the diagnosis and exclusion of endometrial cancer. STUDY DESIGN: All women at risk for endometrial cancer with clinical indications for endometrial biopsy were evaluated by endometrial brush biopsy (Tao Brush, Cook OB-GYN, Bloomington, Indiana) and Pipelle (Cooper Surgical, Shelton, Connecticut) endometrial biopsies during one office visit. Patients were followed longitudinally for the development of endometrial cancer or until undergoing dilatation and curettage or hysterectomy. All comparisons were analyzed using the chi 2 or t test. RESULTS: One hundred one women (mean age, 58; range, 35-86) had endometrial biopsies performed. Median follow-up was > 21 months (range, 3-29). Twenty-two had cancer or atypia, while the remaining had benign diagnoses. When correlated with the final diagnosis, the Tao Brush had 95.5% sensitivity and the Pipelle, 86% sensitivity. Both devices had 100% specificity, positive predictive value of 100% and negative predictive value of 98%. When the results of the two biopsy devices are considered together, the positive and negative predictive value for detecting or excluding endometrial cancer was 100%. Based on 1998 Medicare reimbursements, a simultaneous second office biopsy using the Tao brush could save approximately $67 per case as compared to a sonohistogram and much more when compared to dilatation and curettage. CONCLUSION: Endometrial cancer can be reliably detected and excluded using these two distinct office biopsy devices simultaneously during one office visit. In patients with an indication for endometrial biopsy, no further diagnostic test may be necessary to exclude or diagnose endometrial cancer or atypia.

Role of base plate rotational speed in controlling spheroid size distribution and minimizing oversize particle formation during spheroid production by rotary processing.

The occurrence of material adhesion and formation of oversize particles in the product yield during one-pot spheroid production by rotary processing leads to a less predictable process and a decrease in the usable portion of the total product yield obtained from each production run. The use of variable speeds of the rotating frictional base plate during the spheronization run was investigated for achieving optimal spheroid production. When the base plate speed was increased during liquid addition, the greater centrifugal forces generated improved liquid distribution and the mixing of the moist powder mass, resulting in a decrease in the amount of oversize particles formed. When the base plate was maintained at a high speed throughout the run, the amount of oversize particles and mean spheroid size increased, and a greater "between batch" mean spheroid size variability was also observed. The findings showed that, when higher speeds were used, the residence time must be adjusted accordingly to avoid excessive coalescence and growth while maintaining even liquid distribution. A "low-high-low" speed variation during rotary processing may be used to produce spheroids with a narrow size distribution and with a minimal amount of oversize particles in the total product yield.

Endometrial biopsy using the Tao Brush method. A study of 50 women in a general gynecologic practice.

OBJECTIVE: To test whether the Tao Brush can retrieve sufficient endometrium for diagnosis and also to observe patients' tolerance of its use. STUDY DESIGN: Fifty women underwent Tao Brush sampling: 25 were sampled by Tao Brush alone, and the remainder were sampled by Pipelle immediately following. Patients' reactions to each method were observed. RESULTS: Pipelle currettes larger pieces of endometrium; the Tao Brush obtains smaller pieces. There was no discrepancy between Tao Brush and Pipelle except that Pipelle sampled two of five endometrial polyps, while the Tao Brush sampled none. There was less tissue insufficient for diagnosis with the Tao Brush (2%) than Pipelle (12%). Most patients did not show signs of distress during Tao Brush sampling but grimaced during Pipelle suction curettage. For each of the 16 patients in the second group, the Tao Brush was significantly less painful than Pipelle (P < .01). CONCLUSION: Our data suggest that the Tao Brush is an effective alternative endometrial sampler, causes less pain and produces less tissue insufficient for diagnosis than does the Pipelle. The CPT billing codes (58100) are the same for both the Pipelle and Tao Brush.

Mixed polymer coating for modified release from coated spheroids.

Modified-release drug spheroids coated with an aqueous mixture of high-viscosity hydroxypropylmethylcellulose (HPMC) and sodium carboxymethylcellulose (NaCMC) were formulated. The preparation of core drug spheroids and the coating procedures were performed using the rotary processor and a bottom-spray fluidized bed, respectively. Dissolution studies indicated that incorporation of suitable additives, such as poly(vinylpyrrolidone) (PVP) and poly(ethylene glycol) 400 (PEG) improved the flexibility and integrity of the coat layer by retarding the drug release. An increase in coating levels applied generally retarded the release rate of the drug. However, the ratio of HPMC to NaCMC in the mixed, plasticized polymeric coat played a more dominant role in determining the dissolution T50% values. The optimal ratio of HPMC to NaCMC for prolonged drug release was found to be 3:1, whereas an increase in the amount of NaCMC in the mixed polymer coat only increased drug release. The synergistic viscosity effect of HPMC and NaCMC in retarding drug release rate was greater in distilled water than in dissolution media of pH 1 and 7.2. Cross-sectional view of the scanning electron micrograph showed that all of the coated spheroids exhibited a well-fused, continuous, and distinct layer of coating film. The drug release kinetics followed a biexponential first-order kinetic model.

Effects of physical properties of PEG 6000 on pellets produced by melt pelletization.

This study examined the effects of polyethylene glycol (PEG) binders of different physical forms and from different commercial sources on size and size distribution of pellets produced. The meltable binders used were PEG 6000 in the form of flakes, and coarse and fine powders, and melt pelletization using lactose 450 M was carried out in an 8-liter high-shear mixer. Binder particle size, molecular weight, tack, and viscosity were determined. The results showed that the size and size distribution of the pellets obtained could not be explained by the binder particle size. The size and size distribution of the pellets were related to the tack and viscosity of the molten binders. PEGs used were labeled as the same nominal molecular weight grade, although their determined molecular weights could be quite different. Differences in tack and viscosity of the molten binders were associated with determined molecular weight of the binders. The melt pelletization process is sensitive to tack and viscosity of the molten binders. When different PEG brands of the same nominal molecular weight or different batches of the same brand are used in melt pelletization, it is important to characterize the tack and viscosity of the binders used. The effects of the physical form of binders on the pellet quality appear to be less important when compared to the influences of tack and viscosity of the molten binder.

Effect of off-bottom clearance on properties of pellets produced by melt pelletization.

This study examines the influence of off-bottom clearance on size and size distribution of pellets produced during melt pelletization at different postmelt impeller speeds and binder concentrations using lactose and polyethylene glycol. Melt pelletization of lactose powder 450 M in an 8-liter highshear mixer, the floor of which was made of polytetrafluoroethylene (PTFE), was carried out with polyethylene glycol 3000 as the meltable binder. Erosion of the PTFE floor of the mixer occurred with time of use and this caused a change in the off-bottom clearance. The findings showed that with a wider clearance, the size distribution of pellets was wider and pellets were much larger. These changes were the results of changes in the mixing intensity and impact frequency of the mass in relation to the eddies formed within the off-bottom clearance. The changes were not associated with the reduction in the circulating material load by entrapment into the off-bottom clearance. In the melt pelletization process, the quality of product was higher if the clearance was kept to a minimum. The off-bottom clearance was best measured at the beginning of each pelletization run because the PTFE floor of the mixer was prone to erosion.

Effect of magnesium stearate on chitosan microspheres prepared by an emulsification-coacervation technique.

Chitosan microspheres were prepared using an emulsification-coacervation technique. The w/o emulsion comprised a mixture of light and heavy liquid paraffins containing sodium dioctyl sulphosuccinate as the oil phase and chitosan solution as the aqueous phase. Pentasodium tripolyphosphate was included as a counterion. The chitosan microspheres obtained showed a high degree of aggregation. This was markedly reduced, by the incorporation of magnesium stearate in the disperse phase. The resultant microspheres were then discrete, spherical with smooth surfaces. Additionally, with an increasing magnesium stearate content, larger-sized microspheres were produced. The DSC analysis data suggested that the magnesium stearate was converted to stearic acid during the preparation process. Chitosan microspheres containing propranolol hydrochloride were similarly prepared, but their surface was convoluted and their shape not well defined. Unlike the microspheres without drug, the size of the drug-loaded microspheres decreased with increasing magnesium stearate content. The release of propranolol hydrochloride from the microspheres was fast, irrespective of the content of magnesium stearate. Drug encapsulation efficiency was enhanced when a greater amount of magnesium stearate was used.

Compact cell blocks. Use for body fluids, fine needle aspirations and endometrial brush biopsies.

OBJECTIVE: To obtain an ideal cell block wherein the maximal number of cells are displayed within the smallest area on the block surface. STUDY DESIGN: Cyto-Rich Red (AutoCyte, Inc., Elon College, North Carolina, U.S.A.) is added to fresh cellular sediment in a centrifuge tube at a ratio of 1:1. After two minutes, three to four drops of plasma and topical thrombin (5,000 U/10 mL) is added. The tube is then gently agitated for two minutes, until a gelatinous clot is obtained. The clot is then slid onto a lens tissue on top of paper towels. The lens tissue is folded once over the clot. By gently squeezing the excess fluid from it through the lens tissue into the paper towels, the clot is transformed into a flat, compact, densely cellular aggregate, which is painted with mercurochrome prior to fixation in formaldehyde. RESULTS: From each of the 495 cases, including 250 body cavity fluids, 170 fine needle aspirates and 75 endometrial brush biopsies, processed with the above protocol, there was a compact cell block containing packed cells or tissue fragments in a clean background devoid of red blood cells. CONCLUSION: The compact cell block is about 10-20% the size of a conventional cell block, yet more cells are on display, thus reducing the need for deeper cuts and screening time while increasing the efficiency of cytodiagnosis. The compact cell block technique is particularly helpful for endometrial brush biopsies.

A review of the endometrial changes in Norplant users.

PIP: Prolonged exposure to levonorgestrel associated with use of Norplant contraceptive implants leads, in many acceptors, to menstrual irregularities. Endometrial biopsies collected from a group of Norplant acceptors with persistent or prolonged breakthrough bleeding indicated this side effect is associated with proliferative-type endometrium. The number of both proliferative and secretory endometrium was markedly reduced. The stroma showed edema, sloughing, and inflammatory infiltration. Despite chronic levonorgestrel exposure, only 31% of stroma showed poorly developed decidualization. There was no association between endometrial findings and levonorgestrel levels. Hysteroscopy revealed markedly engorged superficial vessels in long-term Norplant users. It is hypothesized that progestin-related bleeding abnormalities may be due to the changes in the vascular system (e.g., damaged vessels) and hemostatic factors. Confirmation of this hypothesis requires detection of the presence of hemostatic plugs in patients with and without abnormal bleeding, study of the perivascular expression of tissue factor, measurement of estrogen and progesterone receptors at bleeding and non-bleeding sites, and identification of factors that increase angiogenesis and the fragility of uterine vessels.^ieng

Effect of cellulose derivatives on alginate microspheres prepared by emulsification.

Generally discrete and spherical calcium alginate microspheres with a high drug encapsulation efficiency were readily prepared by an emulsification process. They were found to release drug rapidly. In the present study, co-polymer in the form of cellulose derivatives was added to sodium alginate in an attempt to modify the drug release profiles of the microspheres. The effects of cellulose derivatives on the morphology and drug encapsulation efficiency of the microspheres were also evaluated. The cellulose derivatives increased the degree of agglomeration of the microspheres. Small and spherical microspheres were produced from cellulose derivatives of low viscosity while larger microspheres which tended to be elongated were produced from cellulose derivatives of high viscosity. The drug encapsulation efficiency and the drug release profiles were influenced by the chemical nature of the cellulose derivative as well as its viscosity. The efficiency of drug encapsulation generally increased while the rate of drug release decreased with increasing viscosity of the cellulose derivatives. Less hydrophilic cellulose derivatives such as methyl cellulose and hydroxypropylmethyl cellulose were found to increase the efficiency of encapsulating sulphaguanidine, while more hydrophilic cellulose derivatives such as hydroxypropyl cellulose and carboxymethyl cellulose had the opposite effect. Among the cellulose derivatives used, only hydroxypropyl cellulose retarded the drug release of the microspheres.

Prolonged retention of cross-linked trypsin in calcium alginate microspheres.

Trypsin microencapsulated in a calcium alginate matrix was lost quickly through diffusion when the microspheres were placed in an aqueous medium. This problem was overcome by first reacting trypsin with glutaraldehyde to form cross-linkages and then incorporating the enzyme in the alginate microspheres. The performance of the cross-linked trypsin remained optimal at pH 8 while it was found to be more heat-stable and remained highly active even at 80 degrees C. Esters and amides of L-arginine were preferentially hydrolysed by the enzyme indicating that cross-linking did not adversely affect the conformation of the active site. There was a suppression in enzymatic activity when the microspheres were placed in reaction media with an increasing concentration of organic solvent such as ethanol, acetonitrile or isopropanol. However, when returned to a totally aqueous environment, the enzyme resumed its initial tryptic capability. Such a microencapsulated form of cross-linked enzyme may find application in enzyme replacement therapy, optical resolution of racemic compounds as well as organic synthesis in an aqueous-organic environment.

Deviation of the ratio of drugs in a two-component mixture encapsulated in cellulose phthalate microspheres.

Microspheres were formed when a solution of cellulose phthalate was extruded into 30% glacial acetic acid solution. Sulphonamides entrapped in such microspheres leached into the hardening solution because they dissolved freely in the acetic acid solution. This resulted in poor loading efficiency of the sulphonamides in the microspheres. When mixtures of sulphaguanidine and sulphathiazole in various drug ratios were microencapsulated by this method, the observed drug ratios were found to be markedly changed. This was attributed to the difference in solubility of the two sulphonamides in acid such that their extent of diffusion into the hardening solution was not similar. NSAIDS such as ibuprofen and mefenamic acid which are acidic and more hydrophobic in nature are less soluble in acetic acid. These drugs were retained better in the microspheres during the hardening process and the loading efficiency was consequently improved. In cases where mixture of the NSAIDS were encapsulated, the drug ratios showed little deviation from the theoretical values. This study shows that loading of the CAP microspheres is dependent on the solubility of the drugs in acetic acid. When more than one drug is required to be microencapsulated, the drug ratio may change if the drugs have different solubility in acetic acid.

Citric acid as a plasticizer for spray-dried microcapsules.

An aqueous spray-drying process was used to coat theophylline particles with a cellulose polymer, sodium carboxymethylcellulose (NaCMC) or hydroxypropylmethylcellulose (HPMC). Effect of a plasticizer, citric acid, was studied. Strength of cast films have a direct relationship to the dissolution T50% values of the respective spray-dried products. With varying plasticizer concentrations, changes in interaction between plasticizer and polymer, and in size of crystallized drug crystals, were observed with HPMC films. The plasticizer content also affected the films of NaCMC by bringing about a change in the form and arrangement of the drug. X-ray diffraction analyses of the different spray-dried products revealed a possibility of the predominance of different crystal forms. Spherical spray-dried microcapsules of NaCMC were formed with a 20-30% w/w plasticizer content. With HPMC, as plasticizer concentration was increased there was a corresponding increase in mean size of the spray-dried products. In terms of drug release and formation of spherical particles, a plasticizer concentration of 30% w/w was found to be suitable for both HPMC and NaCMC.

Drug encapsulation in alginate microspheres by emulsification.

A method based on an emulsification process was developed for the production of calcium alginate microspheres. Isopropyl alcohol and acetone, which are strong dehydrating agents, were used to aid in the hardening and drying of the microspheres. However, the amount of drug encapsulated was very low. This was due to the drug being soluble in the dehydrating solvents. In the absence of the solvents a high percentage of drug was encapsulated, and this was further increased by forming the microspheres by phase inversion. It was also found that a suspension of the drug particles was required for effective microencapsulation. The efficiency of drug encapsulation generally increased with the ratio of drug to encapsulating material. The microspheres produced were free-flowing and most of them were smaller than 150 microns.

Interaction of preservatives with macromolecules: Part I--Natural hydrocolloids.

Antibacterial activity of methyl-p-hydroxybenzoate against Ps. aeruginosa was evaluated in the presence of varying concentrations of acacia, tragacanth, sodium alginate, guar gum and carrageenin. All these hydrocolloids reduced the antibacterial activity to varying degrees. Tragacanth and guar gum inhibited the activity to a greater extent than acacia, sodium alginate and carrageenin. Hydrocolloids reduce the antibacterial activity of preservatives in two ways. Interaction of the preservative with hydrophilic macromolecules and subsequent reduction in the availability of preservative appears to be the predominant mechanism by which tragacanth and guar gum reduce the activity of methyl-p-hydroxybenzoate. Acacia, sodium alginate and carrageenin apparently act by offering physical protection to microbial cells from the action of the preservative. It is also probable that these hydrocolloids provide more favourable media for microbial growth thereby increasing the preservative requirement for adequate preservation.

Plasticizers and their effects on microencapsulation process by spray-drying in an aqueous system.

Microencapsulated theophylline particles were prepared by an aqueous spray-drying process using hydroxypropylmethylcellulose. The effect of different plasticizers, triethylcitrate, polyethylene glycol, propylene glycol, glycerin and citric acid, was investigated. Triethylcitrate, a water-insoluble plasticizer, produced a porous honeycomb-like microcapsule wall resulting in rapid drug release. The presence of the plasticizers also influenced crystallization of the drug. The formation of a solid drug dispersion was observed with the addition of citric acid or glycerin. Changes in the pH of liquid feed caused by the plasticizer had an effect on the product dissolution profile, but this was not a major factor. Formation of pores due to leaching of plasticizers during dissolution enhanced drug release. Flow property measurements indicated that the plasticizers also affect the cohesiveness of the spray-dried products. Compared to the microcapsules formed without any plasticizers, propylene glycol, glycerin and citric acid appeared to be beneficial to the microcapsule wall formation, with microcapsules containing citric acid having the slowest drug release.