Anthelmintic resistance of intestinal nematodes to ivermectin and pyrantel in Estonian horses.

There is evidence of resistance in horses to anthelmintic treatment using ivermectin and pyrantel. However, little information is available about the parasites, treatment practices or anthelmintic resistance in the horse population in Estonia. In the present study, we examined 41 trotting and riding horses aged < 3 years from four stables in Estonia. Faecal samples were collected, and horses were selected for treatment if the nematode egg count per gram faeces exceeded 200. Horses (n= 32) that shed strongyle-type eggs were treated with pyrantel, whereas Parascaris equorum-positive animals received ivermectin. Up to 78% of horses required anthelmintic treatment and the efficiency of the anthelmintics was evaluated using a faecal egg count reduction test. Resistance of P. equorum was observed in 50% of horses treated with ivermectin and of strongyles in 27% of horses treated with pyrantel. Ivermectin treatment resulted in a mean reduction of 100% for strongyle eggs and an 89% reduction in P. equorum, and pyrantel-treated horses exhibited an 88% reduction in strongyle eggs. These results are considered to be the first indication of resistance to pyrantel, but further studies of ivermectin resistance are required. According to questionnaires completed by the owners of horses, resistance might be explained by a lack of evidence-based strategies, a strong preference for using ivermectin and possibly a subjective evaluation of the body weight of horses.

Efficacy of salbutamol via Easyhaler unaffected by low inspiratory flow.

The fine particle dose delivered via dry powder inhalers (DPIs) is often affected by the inspiratory flow rate generated during inhalation. This has clinical implications, since the fine particle dose determines the amount of drug reaching the lungs. With Easyhaler DPI the fine particle dose remains relatively constant over the range of inspiratory flow rates from 30-60 l min(-1). The aim of this study was to confirm that clinical efficacy is maintained even at low flow rates by comparing the bronchodilating effect of salbutamol (100 microg) delivered via Easyhaler at a target inspiratory flow of 30 l min(-1) with the same dose of salbutamol via pressurised metered-dose inhaler (pMDI) plus spacer. This was a double-blind, randomized, cross-over study with double-dummy technique. Twenty-one paediatric and adult asthmatic patients completed the study, which was conducted over 2 study days. The main outcome parameter was forced expiratory volume in 1 sec (FEV1). The patients were trained to generate a low peak inspiratory flow rate (PIFR) of 30 l min(-1), and the actual PIFR through Easyhaler was recorded. The average PIFR through Easyhaler was 28.7 l min(-1). The difference in the maximum value of FEV1 (FEV1max) between the treatments after drug inhalation was 0.01 l. The mean of FEV1max was 2.67 l after pMDI plus spacer compared to 2.69 l after Easyhaler. Improvements in FEV1 were clinically significant. No significant differences between treatments were found. A reasonably low inspiratory flow rate through Easyhaler produces an equivalent improvement in lung function to a correctly used pMDI plus spacer. Hence, Easyhaler can be used with confidence in patients who may have difficulty in generating a high inspiratory flow rate, such as children and the elderly.

A lipid carrier with a membrane active component and a small complex size are required for efficient cellular delivery of anti-sense phosphorothioate oligonucleotides.

Anti-sense oligonucleotides are potential therapeutic agents that are used to block protein expression from mRNA. To assess the essential properties for an efficient cellular delivery system of phosphorothioate oligonucleotides (PS-ODNs), different cationic carriers were compared. The carriers were complexed with oligonucleotides at various +/- charge ratios in MES-Hepes buffer. Cationic polymers, polylysines (PLL, mean MWs 4000, 20000, 200000 kDa), polyethyleneimines (PEI, mean MWs 25 and 800 kDa) and fractured sixth-generation polyamidoamine dendrimer (PAMAM) were tested for ODN delivery into a D 407 cell line (human retinal pigment epithelial cells) with stably transfected luciferase gene. Anti-sense ODN was directed against the luciferase gene, and the anti-sense effect was determined using a luminometric method. Lipid-based vehicles included DOTAP, DOTAP/DOPE (1/1 by mol), DOTAP/Chol (1/1 by mol), DOTAP/DOPE/Chol (2/1/1 by mol), DOGS and Cytofectin GS/DOPE (2/1 by mol). Additionally a membrane-active peptide JTS-1 (NH(2) -GLFEALLELLESLWELLLEA-COOH) was added to the complexes containing DOTAP, PEI or PLL. In D 407 and CV-1 cells, the anti-sense effect was seen only with lipid-based carriers with a membrane-active component (DOPE or JTS-1). The polymeric systems were ineffective. The effect of the complexation medium was further studied on CV-1 cells. Complexes were prepared in either water, MES-Hepes buffer or cell growth medium (DMEM). Complexes prepared in water were generally most effective and the greater activity is probably due to the smaller complex size. Complex sizes differed greatly in buffer and DMEM, especially in the case of DOPE containing complexes. In conclusion, lipid carrier with a membrane active component and small complex size are required for an efficient cellular delivery of phosphorothioate oligonucleotides.

Microemulsions for topical delivery of estradiol.

Estradiol has been widely used for the treatment of hormonal insufficiencies. Due to its extensive first pass metabolism after oral administration, transdermal administration of estradiol in gels and emulsions has been used to improve its bioavailability, prolong activity and to optimize metabolic profile. The purpose of this study was to investigate microemulsions as delivery systems for estradiol. Various o/w microemulsions were used to deliver estradiol across human abdominal skin in vitro. Trasdermal flux of estradiol was determined using Franz-type diffusion cells and the samples were analyzed by high-performance liquid chromatography (HPLC). The permeation data showed that microemulsion formulations increased estradiol flux 200-700-fold over the control, but permeability coefficients were decreased by 5-18 times. The superior transdermal flux of estradiol was due to 1500-fold improvement in solubilization of estradiol by microemulsions. The results suggest that microemulsions are potential vehicles for improved topical delivery of estradiol.

An X-ray diffraction study of the crystalline composition of gallstones.

The crystalline composition of gallstones removed from 30 patients from southwestern Finland was determined by the X-ray powder method. A total of eight crystalline compounds, varying from one to four per stone, were identified. Anhydrous cholesterol was by far the most abundant compound, occurring in 29 patients (97%), and calcium salts occurred in half the material studied. The stones could be classified on the basis of crystalline composition: pure cholesterol stones (40%), stones of cholesterol and calcium carbonate (37%), stones of cholesterol and sodium chloride or/and calcium palmitate (20%), and a stone of apatite and calcium carbonate (3%). The average amount of crystalline components per stone was as follows: cholesterols, 82%; calcium carbonates, 14%; and the rest, apatite, calcium palmitate, and sodium chloride. The crystalline composition of the stones related to the sex and age of the patients indicated several trends, including the occurrence of calcium carbonates in the stones of patients over 50 years of age and their simultaneous occurrence in small stones and with the cholesterols. Calcium palmitate was also more frequently present in the calculi of male patients.

Randomised trial of the effect of co-administration with acellular pertussis DTP vaccine on immunogenicity of Haemophilus influenzae type b conjugate vaccine.

BACKGROUND: Inclusion of new vaccines in vaccination programmes for children would be easier if they could be combined with existing vaccines. Vaccines containing acellular pertussis in the diphtheria/tetanus/pertussis (DTP-a) combination are expected to replace the conventional whole-cell vaccines (DTP-w). We tested the immunogenicity and safety of a combination of DTP-a with the Haemophilus influenzae type b (Hib) conjugate of Hib capsular polysaccharide and tetanus toxid (PRP-T), and inactivated poliovirus vaccine (i.p.v.). METHODS: 120 infants were enrolled and randomised to four groups to receive DTP-a at ages 2, 4, and 6 months. At 4 and 6 months they also received Hib conjugate and i.p.v., either as separate injections or mixed with DTP-a. All injections were given intramuscularly in the anterolateral area of the thigh. Any reactions after each vaccination were noted by the parents. EIA was used to measure titres of diphtheria, tetanus, and pertussis antibodies, RIA for Hib anticapsular antibodies, and microneutralisation assay for poliovirus antibodies from serum samples collected at the ages of 2, 4, 6, and 7 months. FINDINGS: There were 30 infants in each group. Only mild adverse events were reported. There was a tendency towards slightly lower concentrations of filamentous haemagglutinin, tetanus, and poliovirus 1 antibodies when the vaccines were mixed. However, there was a more pronounced difference (p = 4 x 10(-6)) in Hib antibodies between groups receiving Hib capsular polysaccharide mixed with DTP-a (geometric mean concentrations 0.37 microgram/mL and 0.56 microgram/mL) compared with groups receiving the vaccines separately (3.10 micrograms/mL and 3.94 micrograms/mL). INTERPRETATION: Administration of premixed DTP-a, Hib conjugate, and i.p.v. affect the immune response significantly. The mechanism of this interference is not clear. The immunogenicity of all antigens must be tested before new combinations can be accepted for vaccination programmes for infants.