ABSTRACT
The incidence of fungal pulmonary infections is known to be on the increase, and yet there is an alarming gap in terms of marketed antifungal therapies that are available for pulmonary administration. Amphotericin B (AmB) is a highly efficient broad-spectrum antifungal only marketed as an intravenous formulation. Based on the lack of effective antifungal and antiparasitic pulmonary treatments, the aim of this study was to develop a carbohydrate-based AmB dry powder inhaler (DPI) formulation, prepared by spray drying. Amorphous AmB microparticles were developed by combining 39.7 % AmB with 39.7 % γ-cyclodextrin, 8.1 % mannose and 12.5 % leucine. An increase in the mannose concentration from 8.1 to 29.8 %, led to partial drug crystallisation. Both formulations showed good in vitro lung deposition characteristics (80 % FPF < 5 µm and MMAD < 3 µm) at different air flow rates (60 and 30 L/min) when used with a DPI, but also during nebulisation upon reconstitution in water.
Subject(s)
Amphotericin B , Pneumonia , Humans , Amphotericin B/pharmacology , Antifungal Agents/pharmacology , Dry Powder Inhalers , Mannose , Lung , Macrophages, Alveolar , Administration, Inhalation , Particle Size , Powders/pharmacology , Respiratory Aerosols and DropletsABSTRACT
OBJECTIVE: The focus of this study was to investigate the possibility of producing ibuprofen-nicotinamide (IBU-NIC) and ibuprofen-isonicotinamide (IBU-INA) cocrystal-containing granules, using a one-step fluidized bed dryer granulation manufacturing process, and evaluate their mechanical properties. SIGNIFICANCE: Pharmaceutical cocrystals represent a suitable strategy to improve properties of active pharmaceutical ingredients (APIs), such as solubility and processability. Ibuprofen (IBU) is a small molecule API which can form cocrystals with different coformers, including NIC and INA. An improvement in mechanical properties for IBU-NIC cocrystals relative to IBU was previously reported but, to date, the formulation of IBU cocrystals in a solid dosage form has not been investigated. METHODS: In situ cocrystallization and granulation were achieved concurrently by processing in a lab-scale fluidized bed granulator following a design of experiment (DoE) approach using a two-level factorial design with both process and formulation variables. Solid-state, micrometric, dissolution, and mechanical (tabletability) characteristics of granules were assessed post-processing. RESULTS: Granules containing cocrystals were successfully prepared for 11 of 16 DoE runs. Parameters with a significant effect on granule drug loading, flow function, porosity, and size could be identified from the DoE model. Process yield was increased by using a high inlet temperature at high solution feed rate. To avoid the formation of sticky particles, caking and over-wetting of the powder during the process, the utilization of high inlet temperature, low API + coformer:filler ratio, low API concentration in solution and low solution feed rate were suggested by the model. CONCLUSION: The multivariable model developed enables accurate optimization of the granulation process for IBU cocrystals.
Subject(s)
Excipients , Ibuprofen , Porosity , Powders , SolubilityABSTRACT
Currently in the pharmaceutical industry, continuous manufacturing is an area of significant interest. In particular, hot-melt extrusion (HME) offers many advantages and has been shown to significantly reduce the number of processing steps relative to a conventional product manufacturing line. To control product quality during HME without process interruption, integration of inline analytical technology is critical. Vibrational spectroscopy (Raman, NIR and FT-IR) is often employed and used for real-time measurements because of the non-destructive and rapid nature of these analytical techniques. However, the establishment of reliable Process Analytical Technology (PAT) tools for HME of thermolabile drugs is challenging. Indeed, the Raman effect is inherently weak and might be subject to interference. Moreover, during HME, heating and photodecomposition can occur and disrupt spectra acquisition. The aim of this research article was to explore the use of inline Raman spectroscopy to characterise a thermolabile drug, ramipril (RMP), during continuous HME processing. Offline measurements by HPLC, LC-MS and Raman spectroscopy were used to characterise RMP and its main degradation product, ramipril-diketopiperazine (RMP-DKP, impurity K). A set of HME experiments together with inline Raman spectroscopic analyses were performed. The feasibility of implementing inline Raman spectroscopic analysis to quantify the level of RMP and RMP-DKP in the extrudate was addressed. Two regions in the Raman spectrum were selected to differentiate RMP and RMP-DKP. When regions were combined, a principle component analysis (PCA) model defined by these two main components (PC 1â¯=â¯50.1% and PC 2â¯=â¯45%) was established. Using HPLC analyses, we were able to confirm that the PC 1 score was attributed to the level of RMP-DKP, and the PC 2 score was related to the RMP drug content. Investigation of the PCA scatterplot indicated that HME processing temperature was not the only factor causing RMP degradation. Additionally, the plasticiser content, feeding speed and screw rotating speed contributed to RMP degradation during HME processing.
Subject(s)
Hot Melt Extrusion Technology , Quality Control , Spectrum Analysis, Raman/methods , Chromatography, High Pressure Liquid , Citrates/chemistry , Drug Combinations , Plasticizers/chemistry , Polymethacrylic Acids/chemistry , Ramipril/chemistryABSTRACT
The present study evaluates the effect of L-leucine concentration and operating parameters of a laboratory spray dryer on characteristics of trehalose dry powders, with the goal of optimizing production of these powders for inhaled drug delivery. Trehalose/L-leucine mixtures were spray dried from aqueous solution using a laboratory spray dryer. A factorial design of experiment (DoE) was undertaken and process parameters adjusted were: inlet temperature, gas flow rate, feed solution flow rate (pump setting), aspiration setting and L-leucine concentration. Resulting powders were characterised in terms of particle size, yield, residual moisture content, and glass transition temperature. Particle size was mainly influenced by gas flow rate, whereas product yield and residual moisture content were found to be primarily affected by inlet temperature and spray solution feed rate respectively. Interactions between a number of different process parameters were elucidated, as were relationships between different responses. The leucine mass ratio influenced the physical stability of powders against environmental humidity, and a high leucine concentration (30% w/w) protected amorphous trehalose from moisture induced crystallization. High weight ratio of leucine in the formulation, however, negatively impacted the aerosol performance. Thus, in terms of L-leucine inclusion in a formulation designed for pulmonary delivery, a balance needs to be found between physical stability and deposition characteristics.
Subject(s)
Drug Compounding/methods , Leucine/chemistry , Trehalose/chemistry , Administration, Inhalation , Aerosols , Desiccation , Drug DesignABSTRACT
The purpose of this work was to investigate the application of different advanced continuous processing techniques (hot melt extrusion and spray drying) to the production of fixed-dose combination (FDC) monolithic systems comprising of hydrochlorothiazide and ramipril for the treatment of hypertension. Identical FDC formulations were manufactured by the two different methods and were characterised using powder X-ray diffraction (PXRD) and modulated differential scanning calorimetry (mDSC). Drug dissolution rates were investigated using a Wood's apparatus, while physical stability was assessed on storage under controlled temperature and humidity conditions. Interestingly both drugs were transformed into their amorphous forms when spray dried, however, hydrochlorothiazide was determined, by PXRD, to be partially crystalline when hot melt extruded with either polymer carrier (Kollidon® VA 64 or Soluplus®). Hot melt extrusion was found to result in significant degradation of ramipril, however, this could be mitigated by the inclusion of the plasticizer, polyethylene glycol 3350, in the formulation and appropriate adjustment of processing temperature. The results of intrinsic dissolution rate studies showed that hot-melt extruded samples were found to release both drugs faster than identical formulations produced via spray drying. However, the differences were attributable to the surface roughness of the compressed discs in the Wood's apparatus, rather than solid state differences between samples. After a 60-day stability study spray dried samples exhibited a greater physical stability than the equivalent hot melt extruded samples.
Subject(s)
Angiotensin-Converting Enzyme Inhibitors/chemistry , Antihypertensive Agents/chemistry , Diuretics/chemistry , Hot Temperature , Hydrochlorothiazide/chemistry , Ramipril/chemistry , Technology, Pharmaceutical/methods , Calorimetry, Differential Scanning , Crystallography, X-Ray , Drug Carriers , Drug Combinations , Drug Compounding , Drug Liberation , Drug Stability , Kinetics , Microscopy, Electron, Scanning , Particle Size , Plasticizers/chemistry , Polyethylene Glycols/chemistry , Polyvinyls/chemistry , Powder Diffraction , Pyrrolidines/chemistry , Solubility , Surface Properties , Vinyl Compounds/chemistryABSTRACT
Using the Interactive Autism Network Research Database, the Autistic Spectrum Disorder concordance rates for twins and non-twin siblings were calculated. For males, females and both genders together, the concordance rate for dizygotic twins is approximately twice that of non-twin siblings. We also determined that the concordance rate for non-twin siblings decreases as the interval between pregnancies increases. Our results appear to indicate that the uterine environment may contribute to autism concordance rates.
Subject(s)
Birth Intervals/statistics & numerical data , Child Development Disorders, Pervasive/epidemiology , Diseases in Twins/epidemiology , Siblings , Twins , Uterus/physiology , Female , Humans , Male , Models, BiologicalABSTRACT
Pulmonary administration of protein and peptide drugs using inhaled dry powder particles is an interesting alternative to parenteral delivery. The stabilisation of these molecules is essential to the maintenance of biological activity in such inhalation formulations. Here salmon calcitonin (sCT) was co-spray dried with linear or branched PEG (L-PEG and B-PEG) and PVP in order to formulate aerosolisable particles of the bioactive peptide. Co-spray drying L-PEG and PVP resulted in porous particles, with minimal D(50) (median volume diameter) and MMAD (mass median aerodynamic diameter) values obtained for a PEG/PVP w/w ratio of 1. For particles based on both L-PEG and B-PEG, an increase in acetone, a poor solvent for the PVP, up to 70wt% of the spray dried solution led to a decrease in D(50) and MMAD. Crystallinity of PEG in the particles ranged between 90 and 97% when the PVP content varied between 15 and 70wt%, indicating a low degree of interaction between PVP and PEG. Additionally, dynamic vapour sorption analysis showed that an increase in PVP content increased the particle surface hygroscopicity. Hence, particle properties were adjusted by altering the water/acetone and PEG/PVP ratio in the spray dried solutions. PVP present at the particles surface protects them from melting during the spray drying process but also increases their hygroscopicity, adversely affecting their aerodynamic properties. Targeting a 5wt% of sCT loading resulted in a loading efficiency of 77.9 and 83.6% with L-PEG and B-PEG-based particles, respectively. Loading of sCT in L-PEG or B-PEG-based particles modified particle roughness and D(50), leading to an increase in MMAD of the L-PEG-based particles. However, particles were still considered to be suitable for aerosolisation as their FPFs (fine particle fractions) were higher than 30%. These particles formulated with PVP and PEG allowed sCT biological activity to be maintained when evaluated by measuring cAMP production by T47D cells.
Subject(s)
Lung/metabolism , Polymers/chemistry , Calorimetry, Differential Scanning , Particle Size , Polymers/administration & dosage , Solubility , Surface Tension , Thermogravimetry , X-Ray DiffractionABSTRACT
The objective of this work was to examine the physicochemical properties of spray dried budesonide, formoterol fumarate and their mixtures at two different weight ratios: 100:6 and 400:6 of budesonide and formoterol fumarate, respectively. A comparison of the thermal properties, crystalline/amorphous nature and particle size of the starting micronised as well as processed materials was carried out. The micronised drugs on their own and the physical mixtures were crystalline in contrast to the spray dried counterparts which were shown to be amorphous. The glass transition temperatures (T(g)s) of the processed actives were determined and appeared at 89.5 and 88 degrees C for budesonide and formoterol fumarate, respectively. As for the spray dried composites, an indication of miscibility and/or interactions between the components was indicated by differential scanning calorimetry and infrared analysis. The spray drying in all cases resulted in smooth, spherical microparticles of sizes suitable for inhalation.
Subject(s)
Anti-Asthmatic Agents/chemistry , Budesonide/chemistry , Drug Compounding/methods , Ethanolamines/chemistry , Calorimetry, Differential Scanning , Crystallization , Drug Combinations , Formoterol Fumarate , Microscopy, Electron, Scanning , Particle Size , Phase Transition , Surface Properties , X-Ray DiffractionABSTRACT
The aim of this study was to investigate the effect of changing spray drying parameters on the production of a budesonide/formoterol fumarate 100:6 (w/w) composite. The systems were spray dried as solutions from 95% ethanol/5% water (v/v) using a Büchi 191-Mini Spray Dryer. A 2(5-1) factorial design study was undertaken to assess the consequence of altering spray drying processing variables on particle characteristics. The processing parameters that were studied were inlet temperature, spray drier airflow rate, pump rate, aspirator setting and feed concentration. Each batch of the resulting powder was characterised in terms of thermal and micromeritic properties as well as an in vitro deposition by twin impinger analysis. Overall, the parameter that had the greatest influence on each response investigated was production yield - airflow (higher airflow giving greater yields), median particle size - airflow (higher airflow giving smaller particle sizes) and Carr's compressibility index - feed concentration (lower feed concentration giving smaller Carr's indices). A six- to seven-fold difference in respirable fraction can be observed by changing the spray drying process parameters. The co-spray dried composite system which displayed best in vitro deposition characteristics, showed a 2.6-fold increase in respirable fraction in the twin impinger experiments and better dose uniformity compared with the physical mix of micronised powders.
Subject(s)
Anti-Asthmatic Agents/chemistry , Budesonide/chemistry , Drug Compounding/methods , Drug Compounding/statistics & numerical data , Ethanolamines/chemistry , Calorimetry, Differential Scanning , Crystallization , Drug Combinations , Formoterol Fumarate , Microscopy, Electron, Scanning , Particle Size , Phase Transition , Research Design , Surface Properties , X-Ray DiffractionABSTRACT
The purpose of this study was to prepare excipient-free porous microparticles of bendroflumethiazide by spray drying and to characterise the physicochemical properties of the particles produced. Solutions of bendroflumethiazide in ethanol/water, ethanol/water/ammonium carbonate or methanol/water/ammonium carbonate were spray dried using a laboratory spray dryer. Spray dried products were characterised by scanning electron microscopy, X-ray powder diffraction, differential scanning calorimetry, FTIR, laser diffraction particle sizing and density measurement. Nanoporous microparticles (NPMPs) were prepared from the alcoholic solutions containing ammonium carbonate. NPMPs were amorphous in nature, had median particles sizes less than 3 microm and densities that were significantly reduced compared to non-porous spray dried bendroflumethiazide powder. The novel process may be used to produce excipient-free amorphous microparticles with desirable physical properties such as amorphous solid state, porosity and low bulk density. This new engineering technology has applications in the design of other therapeutic agents such as those used in pulmonary delivery.
Subject(s)
Antihypertensive Agents/chemistry , Bendroflumethiazide/chemistry , Nanoparticles , Antihypertensive Agents/administration & dosage , Bendroflumethiazide/administration & dosage , Desiccation , Microscopy, Electron, Scanning , Nebulizers and Vaporizers , Particle Size , Powders , Spectroscopy, Fourier Transform Infrared , Surface-Active Agents , X-Ray DiffractionSubject(s)
Animal Welfare , Anti-Bacterial Agents/therapeutic use , Cattle Diseases/prevention & control , Dairying/methods , Veterinary Medicine/methods , Animals , Cattle , Cattle Diseases/drug therapy , Cattle Diseases/therapy , Drug Utilization , Female , Ireland , Mastitis, Bovine/drug therapy , Mastitis, Bovine/prevention & control , Mastitis, Bovine/therapyABSTRACT
Somatic cell count (SCC) data for 480 cows in 10 Irish dairy herds from January 2001 until June 2002 were analysed. Herds were selected on the basis of a recent or ongoing history of clinical or subclinical mastitis. An individual cow SCC of 200 000 cells per ml was used as the threshold for elevation of SCC. The duration of elevated SCC prior to drying-off and the magnitude of the elevation in SCC were found to have an impact on the response to dry cow therapy (DCT). A trend also emerged indicating that increasing parity had a negative influence on the response to DCT.
Subject(s)
Mastitis, Bovine/prevention & control , Milk/cytology , Animals , Cattle , Cell Count/veterinary , Female , Ireland , Multivariate Analysis , Parity , PregnancyABSTRACT
This study compared concentrations of amyloid A in bovine milk with the cell-based indicators of intramammary inflammation, somatic cell count and California Mastitis Test. Mammary quarter data pertaining to 180 cows were categorised according to concentrations of serum amyloid A in the cow of origin. Ranked correlation, ranked regression and receiver operator characteristics all demonstrated acceptable agreement between milk amyloid A concentrations and cell-based indices. There were some indications of reduction in this agreement, in cows with raised concentrations of serum amyloid A. However, there were also indications that serum amyloid A did not significantly influence milk amyloid A. The results of the current study indicate that milk amyloid A exhibits good correlation with established cell-based indicators of intramammary inflammation.
Subject(s)
Mammary Glands, Animal/pathology , Mastitis, Bovine/blood , Mastitis, Bovine/pathology , Milk/chemistry , Serum Amyloid A Protein/analysis , Animals , Biomarkers/analysis , Biomarkers/blood , Cattle , Female , Inflammation/blood , Inflammation/diagnosis , Inflammation/pathology , Mastitis, Bovine/diagnosis , Sensitivity and SpecificityABSTRACT
The aim of this work was to investigate the dissolution rate from both the curved and planar surfaces of cylindrical compacts of benzoic acid, which were placed centrally and non-centrally at the base of the vessel of the paddle dissolution apparatus. The effect of fixing the compacts to a particular position on the variability of dissolution results was also examined. In addition, computational fluid dynamics (CFD) was used to simulate fluid flow around compacts in the different positions in the vessel, and the relationship between the local hydrodynamics in the region of the compacts and the dissolution rate determined. The dissolution rate was found to increase from the centre position to the off-centre positions for each surface examined. There was a corresponding increase in maximum fluid velocities calculated from the CFD fluid flow simulations at a fixed distance from the compact. There was less variability in dissolution from compacts fixed to any of the positions compared with those that were not fixed. Fluid flow around compacts in different positions could be successfully modelled, and hydrodynamic variability examined, using CFD. The effect of asymmetric fluid flow was evident visually from the change in shape of the eroded compacts.
Subject(s)
Computer Simulation , Tablets/chemistry , Technology, Pharmaceutical/methods , Benzoic Acid/chemistry , Benzoic Acid/pharmacokinetics , Equipment Design , Rheology/methods , Solubility , Tablets/pharmacokinetics , Technology, Pharmaceutical/instrumentationSubject(s)
Scrapie/epidemiology , Animals , Data Collection , Female , Humans , Ireland/epidemiology , Male , Prevalence , Random Allocation , Scrapie/diagnosis , Sheep , Surveys and QuestionnairesSubject(s)
Acid-Base Equilibrium , Babesiosis/urine , Babesiosis/veterinary , Cattle Diseases/blood , Cattle Diseases/urine , Animals , Babesiosis/blood , Cattle , Female , Hydrogen-Ion Concentration , Ixodes , MaleABSTRACT
The aim of the present study was to determine the apparent diffusion boundary layer and dissolution rate constant for various surfaces of compacts and at various locations in the USP paddle dissolution apparatus. Benzoic acid compacts were coated with paraffin wax leaving only the surface under investigation free for dissolution. The dissolution rates for various surfaces at varying locations in the paddle dissolution vessel were determined from the slope of the dissolution profile (amount dissolved (mg) versus time (min)). The apparent diffusion boundary layer and dissolution rate constant were calculated and were found to vary depending on the surface of the compact from which dissolution took place and also on the location and size of the compact. It may be concluded that, in developing models to describe the dissolution from solid dosage forms, it is not accurate to assume constant hydrodynamics and mass transfer rates at all surfaces of the system, or in different locations within the test device. A more exact description of the hydrodynamics would be necessary in order to precisely model drug dissolution in the paddle dissolution apparatus.
Subject(s)
Technology, Pharmaceutical/instrumentation , Antifungal Agents , Benzoic Acid , Equipment Design , Sensitivity and SpecificityABSTRACT
We examined Gas 6-Axl interactions in human pulmonary artery endothelial cells (HPAEC) and in Axl-transduced HPAEC to test Gas 6 function during endothelial cell survival. We identified the 5.0-kb Axl, 4.2-kb Rse, and 2.6-kb Gas 6 mRNAs in HPAEC. Immunoprecipitation and Western blotting confirmed the presence of these proteins. Gas 6 is present in cell-associated and secreted fractions of growth-arrested HPAEC, independent of cell density. In addition, the Axl receptor is constitutively phosphorylated in growth-arrested cultures, and exogenous Gas 6 enhanced Axl phosphorylation threefold. Gas 6 added to growth-arrested HPAEC resulted in a significant increase in cell number (1.5 nM Gas 6 increased cell number 35%). Flow cytometry revealed that Gas 6 treatment resulted in 28% fewer apoptosing cells. Transduction of a full-length Axl cDNA into HPAEC resulted in 54% fewer apoptosing cells after Gas 6 treatment. Collectively, the data demonstrate antiapoptotic activities for Gas 6 in HPAEC and suggest that Gas 6 signaling may be relevant to endothelial cell survival in the quiescent environment of the vessel wall.
Subject(s)
Endothelium, Vascular/metabolism , Intercellular Signaling Peptides and Proteins , Oncogene Proteins/metabolism , Proteins/metabolism , Pulmonary Artery/metabolism , Receptor Protein-Tyrosine Kinases/metabolism , Animals , Apoptosis , Blotting, Northern , Blotting, Western , Cell Count , Cell Division/drug effects , Cell Line , Cell Survival/drug effects , Culture Media, Serum-Free/pharmacology , Dose-Response Relationship, Drug , Endothelium, Vascular/cytology , Humans , Lung/cytology , Lung/metabolism , Mice , Oncogene Proteins/genetics , Phosphorylation , Precipitin Tests , Proteins/genetics , Proteins/pharmacology , Proto-Oncogene Proteins , Pulmonary Artery/cytology , RNA, Messenger/metabolism , Receptor Protein-Tyrosine Kinases/genetics , Transduction, Genetic , Axl Receptor Tyrosine KinaseABSTRACT
We used whole lung cultures as a model to study blood vessel formation in vitro and to examine the role that epithelial-mesenchymal interactions play during embryonic pulmonary vascular development. Mouse lungs were isolated at embryonic day 11.5 (E11.5) and cultured for up to 4 days prior to blood vessel analysis. Platelet endothelial cell adhesion molecule-1 (PECAM/CD31) and thrombomodulin (TM/CD141) immunolocalization demonstrate that vascular development occurs in lung cultures. The vascular structures identified in lung cultures first appear as a loosely associated plexus of capillary-like structures that with time surround the airways. To investigate the potential role of vascular endothelial cell growth factor (VEGF) during pulmonary neovascularization, we immunolocalized VEGF in embryonic lungs. Our data demonstrate that VEGF is uniformly present in the airway epithelium and the subepithelial matrix of E11.5 lungs. At later time points, E13.5 and E15.5, VEGF is no longer detected in the proximal airways, but is restricted to the branching tips of airways in the distal lung. RT-PCR analysis reveals that VEGF(164) is the predominant isoform expressed in lung cultures. Grafting heparin-bound VEGF(164) beads onto lung explants locally stimulates a marked neovascular response within 48 hr in culture. Semi-quantitative RT-PCR reveals an 18% increase in PECAM mRNA in VEGF(164)-treated whole lung cultures as compared with untreated cultures. The restricted temporal and spatial expression of VEGF suggests that matrix-associated VEGF links airway branching with blood vessel formation by stimulating neovascularization at the leading edge of branching airways.
Subject(s)
Endothelial Growth Factors/metabolism , Lung/embryology , Lung/metabolism , Lymphokines/metabolism , Neovascularization, Physiologic , Animals , Base Sequence , Culture Techniques , DNA Primers/genetics , Endothelial Growth Factors/genetics , Endothelial Growth Factors/pharmacology , Gene Expression Regulation, Developmental , Immunohistochemistry , Lung/blood supply , Lymphokines/genetics , Lymphokines/pharmacology , Mice , Models, Biological , Neovascularization, Physiologic/drug effects , Platelet Endothelial Cell Adhesion Molecule-1/genetics , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Protein Isoforms/genetics , Protein Isoforms/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Thrombomodulin/metabolism , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth FactorsABSTRACT
Thirty of 52 pedigree Suffolk lambs (58 per cent) were born with arthrogryposis characterised by bilateral flexion rigidity of the metacarpophalangeal and carpal joints. The recent introduction of a breeding ram was identified as the only significant risk factor in the flock, and embryo transfer was used to test the hypothesis that the arthrogryposis was an inherited disorder associated with the introduction of this ram. Two adult ewes that had previously produced lambs with arthrogryposis by the ram and four of its affected daughters were available as donors, and 20 crossbred ewes were used as recipients. Ten Suffolk-crossed ewes that had no known familial relationship with the ram were also mated by the ram as controls and they produced 10 normal lambs. Following embryo transfer, 19 lambs were born, of which seven were stillborn; arthrogryposis was evident in 10 of the 12 live lambs. Analysis of the data suggested that in the population under study, arthrogryposis was inherited as an autosomal recessive condition.