Digital image analysis protocol for determining the radiocarpal joint space in the rheumatoid arthritic wrist.

This paper describes a simple protocol for measuring the joint space of the rheumatoid arthritic (RA) wrist from projection radiographs. The protocol is implemented using a computer algorithm based upon the Interactive Data Language platform. The computerized algorithm features a user-friendly graphical interface to aid the operator to measure joint space parameters, namely distance and area, of the wrist vertebral morphometry at the radiocarpal region. Dual-energy X-ray absorptiometry (DXA) radiograph of a standard hand and wrist phantom was evaluated using the measurement protocol to determine the accuracy and precision of the protocol. The accuracy, parameterised by the systematic error, returned a mean of 5.20% for distance and is equal to 3.49% for area measurement. The precision of the measurement protocol, parameterised by the coefficient of variation (CV), for distance returned a mean of 1.96%; the CV for area measurement equals 2.1%. Three observers participated to investigate the repeatability (intra-observer) and reproducibility (inter-observer) of the measurement protocol, parameterised by the CV, using DXA radiographs from a healthy volunteer and a RA patient. The inter-observer repeatability for distance measurement for the respective observers returned mean values of 10.9%, 7.7% and 11.4% for the healthy wrist. However, the results revealed improved repeatability for the RA wrist; the CV for the respective observers returned mean values of 7.7% 7.1% and 10.0%. The inter-observer repeatability for area measurement for the respective observers returned mean values of 10.2%, 7.1% and 10.1% for the healthy wrist. However, the results revealed improved repeatability (in two out of the three observers) for the RA wrist; the CV for the respective observers returned mean values of 6.8% 6.5% and 10.8%. Student's t-test analysis of the intra-observer repeatability revealed that the measurements of distance and area were generally not intra-observer sensitive. On the other hand, student's t-test analysis of the inter-observer reproducibility revealed that half of the distance measurements were inter-observer sensitive; whereas the remaining were not. Similar findings were obtained for area measurements. Overall the results reveal that the variabilities in accuracy and precision tests and the repeatability and reproducibility tests were typically 10% or less. These findings, in addition to the versatility and simplicity of the digital image analysis protocol, lend to the potential of using the protocol to complement the acquisition of bone mineral density data derived from DXA for diagnosing the progression of RA in patients.

Investigating cholesterol metabolism and ageing using a systems biology approach.

CVD accounted for 27 % of all deaths in the UK in 2014, and was responsible for 1·7 million hospital admissions in 2013/2014. This condition becomes increasingly prevalent with age, affecting 34·1 and 29·8 % of males and females over 75 years of age respectively in 2011. The dysregulation of cholesterol metabolism with age, often observed as a rise in LDL-cholesterol, has been associated with the pathogenesis of CVD. To compound this problem, it is estimated by 2050, 22 % of the world's population will be over 60 years of age, in culmination with a growing resistance and intolerance to pre-existing cholesterol regulating drugs such as statins. Therefore, it is apparent research into additional therapies for hypercholesterolaemia and CVD prevention is a growing necessity. However, it is also imperative to recognise this complex biological system cannot be studied using a reductionist approach; rather its biological uniqueness necessitates a more integrated methodology, such as that offered by systems biology. In this review, we firstly discuss cholesterol metabolism and how it is affected by diet and the ageing process. Next, we describe therapeutic strategies for hypercholesterolaemia, and finally how the systems biology paradigm can be utilised to investigate how ageing interacts with complex systems such as cholesterol metabolism. We conclude by emphasising the need for nutritionists to work in parallel with the systems biology community, to develop novel approaches to studying cholesterol metabolism and its interaction with ageing.

Mathematically modelling the dynamics of cholesterol metabolism and ageing.

Cardiovascular disease (CVD) is the leading cause of morbidity and mortality in the UK. This condition becomes increasingly prevalent during ageing; 34.1% and 29.8% of males and females respectively, over 75 years of age have an underlying cardiovascular problem. The dysregulation of cholesterol metabolism is inextricably correlated with cardiovascular health and for this reason low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) are routinely used as biomarkers of CVD risk. The aim of this work was to use mathematical modelling to explore how cholesterol metabolism is affected by the ageing process. To do this we updated a previously published whole-body mathematical model of cholesterol metabolism to include an additional 96 mechanisms that are fundamental to this biological system. Additional mechanisms were added to cholesterol absorption, cholesterol synthesis, reverse cholesterol transport (RCT), bile acid synthesis, and their enterohepatic circulation. The sensitivity of the model was explored by the use of both local and global parameter scans. In addition, acute cholesterol feeding was used to explore the effectiveness of the regulatory mechanisms which are responsible for maintaining whole-body cholesterol balance. It was found that our model behaves as a hypo-responder to cholesterol feeding, while both the hepatic and intestinal pools of cholesterol increased significantly. The model was also used to explore the effects of ageing in tandem with three different cholesterol ester transfer protein (CETP) genotypes. Ageing in the presence of an atheroprotective CETP genotype, conferring low CETP activity, resulted in a 0.6% increase in LDL-C. In comparison, ageing with a genotype reflective of high CETP activity, resulted in a 1.6% increase in LDL-C. Thus, the model has illustrated the importance of CETP genotypes such as I405V, and their potential role in healthy ageing.

Cholesterol metabolism: A review of how ageing disrupts the biological mechanisms responsible for its regulation.

Cholesterol plays a vital role in the human body as a precursor of steroid hormones and bile acids, in addition to providing structure to cell membranes. Whole body cholesterol metabolism is maintained by a highly coordinated balancing act between cholesterol ingestion, synthesis, absorption, and excretion. The aim of this review is to discuss how ageing interacts with these processes. Firstly, we will present an overview of cholesterol metabolism. Following this, we discuss how the biological mechanisms which underpin cholesterol metabolism are effected by ageing. Included in this discussion are lipoprotein dynamics, cholesterol absorption/synthesis and the enterohepatic circulation/synthesis of bile acids. Moreover, we discuss the role of oxidative stress in the pathological progression of atherosclerosis and also discuss how cholesterol biosynthesis is effected by both the mammalian target of rapamycin and sirtuin pathways. Next, we examine how diet and alterations to the gut microbiome can be used to mitigate the impact ageing has on cholesterol metabolism. We conclude by discussing how mathematical models of cholesterol metabolism can be used to identify therapeutic interventions.

The association between plumage damage and feather-eating in free-range laying hens.

Severe feather-pecking (SFP) persists as a highly prevalent and detrimental behavioural problem in laying hens (Gallus gallus domesticus) worldwide. The present experiment investigated the association between feather-eating and plumage damage, a consequence of SFP, in groups of free-range, ISA Brown laying hens. Single feathers were placed on the floor of the home pens. Feathers were sourced from seven different birds. A total of 50 birds in six pens with extensive plumage damage were compared with birds in six control pens with little plumage damage at 41 to 43 weeks of age (n=12 pens, 600 hens). Birds in pens with extensive plumage damage ingested more feathers (F=8.1, DF=1, 8, P=0.02), and also showed shorter latencies to peck at (χ 2=54.5, DF=1, P<0.001), and ingest feathers (χ 2=55.6, DF=1, P<0.001). Birds ingested feathers from a bird in the free-range facility, in which the testing took place, more quickly than from a bird housed in a separate cage facility (χ 2 = 39.0, DF=6, P<0.001). A second experiment investigated the predictive relationship between feather-eating and plumage damage. Feathers were presented to 16 pens of 50 pullets prior to the development of plumage damage, at 15 weeks of age, and then to the same hens after plumage damage had become prominent, at 40 weeks of age. Birds had a higher probability of ingesting feathers (F=142.0, DF=1, 231, P<0.001), pecked feathers more times (F=11.24, DF=1, 239, P<0.001), and also pecked (χ 2 = 127.3, DF=1, P<0.001) and ingested (χ 2=189.3, DF=1, P<0.001) the feathers more quickly at 40 than 15 weeks of age. There was a trend for an interaction, where birds pecked feathers from the rump more times than feathers from the back at 40 weeks of age (F=3.46, DF=1, 237, P=0.06). However, a lack of variability in plumage damage between pens in this experiment precluded investigation of the predictive relationship. The results from the present study confirm the association between feather-eating and plumage damage, and suggest that birds may prefer feathers from particular body areas and from particular hens. Future experiments should focus on elucidating whether feather-eating may act as a predictor of SFP.

Relationships between range access as monitored by radio frequency identification technology, fearfulness, and plumage damage in free-range laying hens.

Severe feather-pecking (SFP), a particularly injurious behaviour in laying hens (Gallus gallus domesticus), is thought to be negatively correlated with range use in free-range systems. In turn, range use is thought to be inversely associated with fearfulness, where fearful birds may be less likely to venture outside. However, very few experiments have investigated the proposed association between range use and fearfulness. This experiment investigated associations between range use (time spent outside), fearfulness, plumage damage, and BW. Two pens of 50 ISA Brown laying hens (n=100) were fitted with radio frequency identification (RFID) transponders (contained within silicone leg rings) at 26 weeks of age. Data were then collected over 13 days. A total of 95% of birds accessed the outdoor run more than once per day. Birds spent an average duration of 6.1 h outside each day over 11 visits per bird per day (51.5 min per visit). The top 15 and bottom 15 range users (n=30), as determined by the total time spent on the range over 13 days, were selected for study. These birds were tonic immobility (TI) tested at the end of the trial and were feather-scored and weighed after TI testing. Birds with longer TI durations spent less time outside (P=0.01). Plumage damage was not associated with range use (P=0.68). The small group sizes used in this experiment may have been conducive to the high numbers of birds utilising the outdoor range area. The RFID technology collected a large amount of data on range access in the tagged birds, and provides a potential means for quantitatively assessing range access in laying hens. The present findings indicate a negative association between fearfulness and range use. However, the proposed negative association between plumage damage and range use was not supported. The relationships between range use, fearfulness, and SFP warrant further research.

The effects of environmental enrichment and beak-trimming during the rearing period on subsequent feather damage due to feather-pecking in laying hens.

This experiment investigated effects of environmental enrichment and beak-trimming during the rearing period on behavior in rearing and plumage damage later in life. Treatments were applied in a 2 × 2 factorial arrangement. Half of the birds were beak-trimmed at 1 d of age using an infra-red laser. A follow-up light-trim was performed at 11 wk of age with a hot blade. Environmental enrichment consisted of pecking strings, whole oats in the litter, and greater litter depth. Sixteen pens of 50 ISA Brown laying hens were used. Four pullets were selected from each pen as focal birds and observed in their home pens between 3 and 14 wk of age. Plumage damage was scored at the end of the experiment in wk 43. Beak-trimmed birds performed less ground-pecking (P = 0.003), less severe feather-pecking (P = 0.021) and more gentle feather-pecking (P = 0.018) than their non-trimmed counterparts during the rearing period. These birds also exhibited less feather damage in wk 43 (P < 0.001). The results indicate that gentle feather-pecking during rearing is not related to plumage damage when older. Additionally, higher rates of ground-pecking and severe feather-pecking during rearing may be predictive of plumage damage later in life. There was no effect of enrichment on plumage damage. It was concluded that while there was no effect on enrichment, beak-trimming appeared to be effective in reducing plumage damage in wk 43.

Assessment of postcrumble addition of limestone and calcium-specific appetite in broilers during the starter phase.

A study was done to determine whether broilers can regulate Ca intake when limestone is provided separately or mixed with a crumbled feed of variable Ca and P content, and the influence of this on performance and apparent ileal digestibility (AID) of Ca and P (AIDP). Twelve crumbled diets were fed from 10 to 20 d of age (8 replicates, 8 broilers/replicate). Diets A to D contained 0.28% nonphytate P (nPP) and 0.27, 0.51, 0.77, and 1.02% Ca, respectively. Diets E to H contained 0.48% nPP and 0.41, 0.51, 0.77, and 1.02% Ca, respectively. A large particle size limestone was mixed manually to the crumbled diet on a daily basis to achieve 1.02% total Ca in diets A to H. Diets I to L had the same Ca and nPP as diets A to D, but limestone was provided in a separate feeder to assess spatial importance of limestone supply. Limestone consumption, provided in a separate feeder, decreased as Ca concentration increased in the crumble diet (P < 0.05). Calcium intake increased as Ca concentration in crumbled diets increased (P < 0.05). Increased tibia ash and decreased AIDP were observed as Ca intake increased (P < 0.05). When limestone was added to diets containing 0.28% nPP postcrumble, Ca intake (6.38 g/bird), tibia ash (717 mg/bone), and AIDP (39.78%) were not affected by crumbled diet Ca concentration or consumed Ca. Broilers fed diets containing 0.48% nPP and limestone mixed with the crumble, Ca intake changed (5.96, 6.93, 6.59, and 6.04 g/bird for crumble diet with 0.41, 0.51, 0.77, and 1.02% Ca, respectively). Increasing Ca concentration in the crumble from 0.41 to 1.02% resulted in greater tibia ash (875 mg/bone) but lower AIDP (P < 0.05), although Ca intake was similar. In conclusion, when large particle size limestone was provided ad libitum, the ability of broilers to select for Ca was not sufficient to meet their requirement when crumble Ca was less than 0.77%. The AIDP was highest in birds fed the 0.27% Ca concentration diet.

Effect of dietary nonphytate phosphorus and calcium concentration on calcium appetite of broiler chicks.

The effect of dietary nonphytate P (nPP) and Ca concentration on the Ca appetite in broilers was evaluated. A total of 288 one-day-old male Ross 308 broilers were fed a commercial diet for 7 d then randomly allocated to 1 of 8 dietary treatments for a 28-d study. Diets were corn-soybean meal based and formulated to be nutritionally adequate except for nPP and Ca. Two concentrations of Ca (5.0 and 10.0 g/kg) and 4 of nPP (2.5, 3.5, 4.5, and 5.5 g/kg) were used, and all birds had access to a separate Ca source (CaCO3). Bird performance, nutrient digestibility, and tibia ash were determined. Birds fed 5.0 g of Ca/kg diets consumed more (P < 0.01) of the separate Ca source than birds fed diets containing 10.0 g of Ca/kg. Increased consumption (P < 0.01) of the separate Ca source was associated with increasing nPP concentration. Bird performance was not influenced by dietary treatment. Birds fed 5.5 g of nPP/kg diets had lower (P < 0.01) digestibility of DM, CP, and energy than the other groups. Phosphorus digestibility was reduced in birds fed high Ca diets and those fed 2.5 g of nPP/kg diets (P < 0.001). Birds fed 2.5 g of nPP/kg had lower tibia ash values than those fed higher concentrations of nPP while feeding diets containing 10.0 g of Ca/kg led to higher concentrations (P < 0.05) of tibia ash than for birds fed 5.0 g of Ca/kg. This study confirms previous findings that birds are able to meet their Ca requirement when fed Ca separately from the mixed ration. Consumption of the separate Ca source responded to not only Ca concentration but also to the amount of nPP in the diet. These data suggest that dietary nPP concentration influences the Ca specific appetite of broilers, and this may indicate that birds attempt to regulate their intake of Ca relative to nPP. This may be mediated via a physiological mechanism to maintain an appropriate Ca:nPP intake.

Nutritional geometry of calcium and phosphorus nutrition in broiler chicks. Growth performance, skeletal health and intake arrays.

The interaction between calcium (Ca) and non-phytate phosphorus (nPP) in broiler nutrition and skeletal health is highly complex with many factors influencing their digestion, absorption and utilisation. The use of an investigative model such as the geometric framework allows a graphical approach to explore these complex interactions. A total of 600 Ross 308-day-old male broiler chicks were allocated to one of 15 dietary treatments with five replicates and eight birds per replicate. Dietary treatments were formulated to one of three total densities of total Ca+nPP; high (15 g/kg), medium (13.5 g/kg) and low (12 g/kg) and at each density there were five different ratios of Ca : nPP (4, 2.75, 2.1, 1.5 and 1.14 : 1). Weekly performance data was collected and at the end of the experiment birds were individually weighed and the right leg removed for tibia ash analysis. Skeletal health was assessed using the latency to lie (LTL) at day 27. At low Ca and high nPP as well as high Ca and low nPP diets, birds had reduced feed intake, BW gain, poorer feed efficiency and lower tibia ash, resulting in a significant interaction between dietary Ca and nPP (P<0.05). LTL times were negatively influenced by diets having either a broad ratio (high Ca, low nPP) or too narrow a ratio (low Ca, high nPP) indicating that shorter LTL times may be influenced by the ratio of Ca : nPP rather than absolute concentrations of either mineral. The calculated intake arrays show that broilers more closely regulate Ca intake than nPP intake. Broilers are willing to over consume nPP to defend a Ca intake target more so than they are willing to over consume Ca to defend an nPP target. Overall dietary nPP was more influential on performance metrics, however, from the data it may appear that birds prioritise Ca intake over nPP and broadly ate to meet this requirement. As broilers are more willing to eat to a Ca intake target rather than an nPP intake target, this emphasises the importance of formulating diets to a accurately balanced density of Ca : nPP considering the biological importance of both minerals.

Nutritional geometry of calcium and phosphorus nutrition in broiler chicks. The effect of different dietary calcium and phosphorus concentrations and ratios on nutrient digestibility.

A total of 600 Ross 308-day-old male broiler chicks were used in a 28 day digestibility study to investigate the interaction between dietary calcium (Ca) and non-phytate phosphorus (nPP) on the digestibility of minerals and amino acids. Diets were formulated to be nutritionally adequate except for Ca and nPP. Fifteen mash diets based on corn and soya bean meal with varying concentrations of Ca (6.4 to 12.0 g/kg) and nPP (2.4 to 7.0 g/kg) were used. Diets were clustered around total densities of Ca and nPP of 12, 13.5 or 15.0 (g/kg) and within each density, a range of five Ca : nPP ratios (1.14 : 1, 1.5 : 1, 2.0 : 1, 2.75 : 1 and 4.0 : 1) were fed. Birds had free access to feed and water throughout the study. At day 28, birds were euthanised for the determination of apparent ileal mineral and amino acid digestibility. Data were modelled in R version 2.15 using a linear mixed-effects model and interrogation of the data was performed by fitting a low order polynomial function. At high Ca concentrations, increasing nPP led to an increase in the apparent digestibility of minerals. Apparent ileal digestibility of phosphorus (P) was enhanced with increasing dietary nPP up to 5.5 g/kg beyond which no improvements were found. Maximal Ca digestibility was found in diets with >8.0 g/kg Ca with concomitant low concentrations of nPP. Diets with a broader Ca : nPP ratio improved the digestibility of Ca but were deleterious to the digestibility of P. In this study, apparent digestibility of amino acids was broadly unaffected by dietary Ca and nPP concentrations. However, interactions between Ca and nPP were observed for the digestibility of glutamine, tyrosine and methionine (all P<0.001). Nitrogen digestibility showed discrete optima around 10.0 and 5.0 g/kg nPP and Na digestibility was maximised around 8 to 9.0 g/kg Ca and 4.5 to 5.4 g/kg nPP. These data show that the ratio of Ca : nPP is more influential to mineral digestibility than the absolute dietary concentration of each macro mineral.

Androgen-related expression of G-proteins in ovarian cancer.

BACKGROUND: Epidemiological and in vitro data implicate androgens in the aetiology of ovarian cancer, but the mechanisms by which this is mediated are unclear. In this study, we wished to examine the effects of androgens on gene expression in ovarian cancer. METHODS: The expression of androgen receptor (AR) in OVCAR3 and OSEC2 cells was confirmed using immunoblotting and response to androgens was measured using flow cytometric assessment of S-phase fraction. The differential gene expression between androgen stimulated and unstimulated OVCAR3 ovarian cancer cells was examined with a cDNA microarray. The upregulation of a subset of these genes was then confirmed with reverse transcriptase PCR in both OVCAR3 and OSEC2, an ovarian epithelial cell line. Finally, the clinical significance of this upregulation was investigated by examining the expression of Rab25 and Rab35, two G-protein-related molecules in an ovarian cancer tissue microarray (TMA). RESULTS: OVCAR3 and OSEC2 cells were shown to express the AR and showed an increase in S-phase fraction in response to androgen treatment. Treatment of OVCAR3 cells with androgen resulted in a significant upregulation of 121 genes. These findings were confirmed for a subset of seven monomeric G-protein-related genes in both OVCAR3 and OSEC2 cells. After staining for Rab25 and Rab35, the majority of TMA sections examined showed expression for Rab25 (92%) and Rab35 (95%). The expression of Rab25 correlated with histological grade, and expression was higher in endometrioid (median histoscore 10.5) than serous (7.5) or mucinous (5.3) tumours. The expression of Rab25 correlated positively with AR expression supporting its role as an androgen responsive gene in ovarian cancer. CONCLUSIONS: These results suggest that androgens can effect expression of the oncogenic GTPases in ovarian cancer. We propose that the androgen responsive Rab35 may have clinical importance as a biomarker of AR function.

Expression of gonadotrophin releasing hormone receptor I is a favorable prognostic factor in epithelial ovarian cancer.

The majority of epithelial ovarian cancers originate in the ovarian surface epithelium. The ovarian surface epithelium is a hormonally responsive tissue, and hormones are thought to play a key role in the development of this type of cancer. Gonadotrophin releasing hormone II is one of 2 isoforms which are thought to act through gonadotrophin releasing hormone receptor I, and gonadotrophin releasing hormone II has been shown to cause growth inhibition of cultured ovarian surface epithelium. The aim of this study was to investigate the expression levels and prognostic significance of gonadotrophin releasing hormone II and the gonadotrophin releasing hormone receptor I in epithelial ovarian cancer. Gonadotrophin releasing hormone II and gonadotrophin releasing hormone receptor I messenger RNA expression was examined in 23 cancers and 7 normal ovarian surface epithelium samples by quantitative real time polymerase chain reaction. An ovarian cancer tissue microarray containing 139 cases was constructed and immunohistochemical analysis of gonadotrophin releasing hormone II and gonadotrophin releasing hormone receptor I protein expression was performed and correlated with clinical outcome data. Gonadotrophin releasing hormone II messenger RNA expression was lower in cancer samples compared to normal ovarian surface epithelium samples (P < .05). Gonadotrophin releasing hormone II protein expression correlated with histologic subtype (25% serous versus 45% nonserous, P < .05) but not with overall survival. Gonadotrophin releasing hormone receptor I messenger RNA expression was highest in serous tumors when compared to non serous (P < .05) and normal tissue (P < .001). Expression of the gonadotrophin releasing hormone receptor I protein was also found to correlate with patient survival (P < .05). We have demonstrated gonadotrophin releasing hormone II and its receptor, gonadotrophin releasing hormone receptor I, are present in clinical ovarian samples, and that gonadotrophin releasing hormone receptor I protein expression is a favorable prognostic factor, suggesting these proteins play an important role in the development of epithelial ovarian cancer.

Model fitting in two dimensions to small angle diffraction patterns from soft tissue.

In our research programme small angle x-ray scattering (SAXS) is used to provide information on the axial arrangement of collagen molecules as well as data about the state of other components of the extra cellular matrix (ECM) in human tissues. Derivation of parameters to describe and simplify the data is required for much of the SAXS patterns analysis. A method is presented here to achieve function fitting to collagen diffraction peaks along with a representation of the underlying diffuse scatter. A simple model was used which proved reliable in fitting a variety of 2D diffraction patterns. The logarithm of the scatter intensity over the area of the scatter image was taken to reduce the range and improve fitting accuracy. Our model was then used to fit the log data. The model consisted of a radial exponential diffuse scatter component added to a specified number of Gaussian peaks. In 2D the peak model is toroidal, each component being rotated about a common specified centre. Initial search parameters from a 1D averaged sector were supplied to the iterative 2D fitting routine. With the aid of data weighting and basic wavelet filtering, successful and reliable fitting of a specified 2D model to real data is achievable. The process is easily automated. Multiple SAXS patterns can be fitted without operator intervention. As described the model is simple enough to converge rapidly and yet allows image data to be parameterized to a form suitable for extracting the requisite information. The fitting method is flexible enough to be extended to achieve a more comprehensive and complex pattern fitting in two dimensions if this turns out to be necessary. It is our intention to implement orientation distribution functions in the near future by including an angular scaling factor.

A preliminary study of breast cancer diagnosis using laboratory based small angle x-ray scattering.

Breast tissue collected from tumour samples and normal tissue from bi-lateral mastectomy procedures were examined using small angle x-ray scattering. Previous work has indicated that breast tissue disease diagnosis could be performed using small angle x-ray scattering (SAXS) from a synchrotron radiation source. The technique would be more useful to health services if it could be made to work using a conventional x-ray source. Consistent and reliable differences in x-ray scatter distributions were observed between samples from normal and tumour tissue samples using the laboratory based 'SAXSess' system. Albeit from a small number of samples, a sensitivity of 100% was obtained. This result encourages us to pursue the implementation of SAXS as a laboratory based diagnosis technique.

The development of a blend-sampling technique to assess the uniformity of a powder mixture.

This article discusses the challenges overcome during the development of a blend-sampling technique and the successful validation of the blending operation for a tablet dosage form containing 2% active ingredient. Content uniformity results are discussedfor three pilot-scale (15-kg) and seven commercial-scale (150-kg) batches of tablets. Blend and core content uniformity data from the pilot-scale batches were acceptable. For the initial commercial-scale batches, although the tablet core content uniformity data were acceptable, the blend uniformity results were poor. The blend data for these batches had very high mean values, but acceptable relative standard deviations (RSDs). This suggested that the drug was being preferentially sampled by the thief but in a consistent, reproducible manner. Extensive testing was performed on a commercial-scale development batch to identify potential causes of sampling error. The results of this testing helped define the blend-sampling technique and strategy used to validate the mixing operation.

Spinous process strength.

STUDY DESIGN: Mechanical testing of cadaveric lumbar spines and dual energy radiograph absorptiometry scanning were performed. OBJECTIVES: To devise a technique to measure the strength of lumbar spinous processes and to determine the bone mineral density of the vertebrae used. SUMMARY OF BACKGROUND DATA: The spinous process has been identified as the weakest part of the anatomy to which a flexible fixation device can be attached. It was unknown if the spinous processes could withstand the forces applied by the device. METHODS: A hook was fitted to the spinous process of 32 lumbar vertebrae. A custom-built rig was designed to secure a vertebra to a materials testing machine. A loop of cord was passed over a bar mounted on the crosshead of the machine and around the two bollards of the hook. As the crosshead was raised, a tension was applied to the cord. Each vertebra was tested to failure. The bone mineral density of each vertebra was then measured using dual energy radiograph absorptiometry. RESULTS: Failure of the specimens occurred by failure of the spinous process, pedicles, or vertebral body. The logarithm (base 10) of the load (N) at which failure occurred was 2.53 +/- 0.3, which corresponded to a mean failure load of 339 N. The bone mineral density of each vertebral body varied between 0.263 and 0.997 g/cm2. A significant linear correlation was found between bone strength and bone mineral density (P < 0.0001). CONCLUSIONS: Specimens with a bone mineral density in the range of 0.263-0.997 g/cm2 failed at a mean load of 339 N when the load was applied through the spinous process hook of a flexible fixation device.

Two families with an autosomal dominant inheritance pattern for papillary carcinoma of the thyroid.

BACKGROUND: Papillary carcinoma of the thyroid (PTC) is the most prevalent malignancy of the thyroid gland. Although the majority of lesions are sporadic tumors, an established relationship exists between familial adenomatous polyposis (FAP) and PTC. Moreover, some authors postulate the existence of familial PTC as a distinct entity. Evidence for this is limited, however, there being few well characterized descriptions of pedigrees with high prevalence of PTC. AIMS: The objective of the present study was to examine an apparent heritable predisposition to PTC occurring in two Tasmanian families in which PTC occurs commonly. METHODS: Pedigree charts were constructed for both families and the medical records of the members reviewed. RESULTS: In Pedigree I, 7 of 25 members had PTC (6 of these had coexisting multinodular goiter (MNG), and 11 others had MNG. In Pedigree II, identical male twins and their daughters had PTC. CONCLUSIONS: In both families there is evidence of autosomal dominant inheritance of PTC. The association of PTC with MNG suggests a possible role for MNG in tumor pathogenesis in hereditary PTC. The majority of the patients were diagnosed with PTC before commencement of prospective screening, indicating clinically relevant disease in the families described.

Evidence that different regional sympathetic outflows vary in their sensitivity to the sympathoinhibitory actions of putative 5-HT1A and alpha 2-adrenoceptor agonists in anaesthetized cats.

1. An investigation was carried out to determine whether the centrally acting hypotensive drugs whose mechanisms of action are due either to activation of 5-HT1A receptors (flesinoxan, 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) and urapidil--also an alpha 1-adrenoceptor antagonist) or to activation of alpha 2-adrenoceptors (clonidine and moxonidine) cause differential sympathoinhibition. 2. Cats were anaesthetized with alpha-chloralose and simultaneous recordings were made of whole cardiac, splanchnic and renal nerve activity, blood pressure and heart rate. Cumulative dose-response (i.v.) curves were constructed in separate experiments for the above hypotensive agents on these parameters. 3. Renal nerve activity was found to be more sensitive to the sympathoinhibitory action of flesinoxan and 8-OH-DPAT when compared with cardiac nerve activity, whereas the reverse was observed for clonidine and moxonidine, cardiac being more sensitive than renal nerve activity. Splanchnic nerve activity was similarly affected by all drugs. Furthermore at the highest dose, all drugs tended to cause complete inhibition in all regional sympathetic nerve outflows. 4. Urapidil differed from all the above hypotensive drugs in that it caused a similar degree of sympathoinhibition in all sympathetic outflows at all doses. It is suggested that this may be due to the ability of urapidil to block central alpha 1-adrenoceptors in addition to stimulation of 5-HT1A receptors.