An open-label, phase 2, single centre, randomized, crossover design bioequivalence study of AndroForte 5 testosterone cream and Testogel 1% testosterone gel in hypogonadal men: study LP101.

We compared a novel 5% testosterone (T) cream (AndroForte 5, Lawley Pharmaceuticals, Australia) with a 1% T gel (Testogel, Besins Healthcare, Australia). Using an open-label crossover design, subjects were randomized to one of two treatment sequences using either the T gel or T cream first in a 1 : 1 ratio. Each treatment period was 30 days with a 7-14 days washout period between them. On Days 1 and 30 of each treatment period blood was sampled at -15, -5 min, 0, 2, 4, 5, 6, 7, 8, 9, 10, 12 and 16 h post study drug administration. Sixteen men with established androgen deficiency aged between 29 and 73 years, who had undertaken a washout from prior testosterone therapy participated in the study. One subject failed to complete both arms and another was excluded post-completion because of a major protocol violation. Bioequivalence was established based on key pharmacokinetic (PK) variables: AUC, C(avg), C(max), T(max), % fluctuation (with and without baseline correction) for the two formulations of testosterone on Day 1 and Day 30. The ratio and 90% CI of AUC 0.99 (0.86-1.14), C(max) 1.02 (0.84-1.24) and C(avg) 0.99 (0.86-1.14) for T cream/T gel were within the predetermined bio-equivalence criteria of 80% to 125% at Day 30. There were no statistically significant differences between secondary biochemical markers: serum dihydrotestosterone (DHT), oestradiol (E2), sex hormone-binding globulin (SHBG), luteinizing hormone (LH) and (FSH). The two testosterone formulations were shown to be bioequivalent.

Separate value comparison and learning mechanisms in macaque medial and lateral orbitofrontal cortex.

Uncertainty about the function of orbitofrontal cortex (OFC) in guiding decision-making may be a result of its medial (mOFC) and lateral (lOFC) divisions having distinct functions. Here we test the hypothesis that the mOFC is more concerned with reward-guided decision making, in contrast with the lOFC's role in reward-guided learning. Macaques performed three-armed bandit tasks and the effects of selective mOFC lesions were contrasted against lOFC lesions. First, we present analyses that make it possible to measure reward-credit assignment--a crucial component of reward-value learning--independently of the decisions animals make. The mOFC lesions do not lead to impairments in reward-credit assignment that are seen after lOFC lesions. Second, we examined how the reward values of choice options were compared. We present three analyses, one of which examines reward-guided decision making independently of reward-value learning. Lesions of the mOFC, but not the lOFC, disrupted reward-guided decision making. Impairments after mOFC lesions were a function of the multiple option contexts in which decisions were made. Contrary to axiomatic assumptions of decision theory, the mOFC-lesioned animals' value comparisons were no longer independent of irrelevant alternatives.

Does the medial orbitofrontal cortex have a role in social valuation?

It has been claimed that social behaviour changes after lesions of the ventromedial prefrontal cortex (vmPFC). However, lesions in humans are rarely restricted to a well defined cortical area. Although vmPFC lesions usually include medial orbitofrontal cortex (mOFC), they typically also affect subgenual and/or perigenual anterior cingulate cortex. The purpose of the current study is to investigate the role of mOFC in social valuation and decision-making. We tested four macaque monkeys prior to and after focal lesions of mOFC. Comparison of the animals' pre- and postoperative performance revealed that, unlike lesions of anterior cingulate gyrus (ACCg), lesions of mOFC did not induce alterations in social valuation. MOFC lesions did, however, induce mild impairments in a probabilistic two-choice decision task, which were not seen after ACCg lesions. In summary, the double dissociation between the patterns of impairment suggest that vmPFC involvement in both decision-making and social valuation may be mediated by distinct subregions centred on mOFC and ACCg respectively.

A role for the macaque anterior cingulate gyrus in social valuation.

Complex human social interaction is disrupted when the frontal lobe is damaged in disease, and in extreme cases patients are described as having acquired sociopathy. We compared, in macaques, the effects of lesions in subdivisions of the anterior cingulate and the orbitofrontal cortices believed to be anatomically homologous to those damaged in such patients. We show that the anterior cingulate gyrus in male macaques is critical for normal patterns of social interest in other individual male or female macaques. Conversely, the orbitofrontal cortex lesion had a marked effect only on responses to mildly fear-inducing stimuli. These results suggest that damage to the anterior cingulate gyrus may be the cause of changes in social interaction seen after frontal lobe damage.

Endoscopic argon plasma coagulation for the treatment of gastric antral vascular ectasia (watermelon stomach): long-term results.

INTRODUCTION: Gastric antral vascular ectasia is a rare but well-recognised cause of occult gastrointestinal bleeding. Various endoscopic treatments have been tried in this condition. We report our experience with argon plasma coagulation in the treatment of gastric antral vascular ectasia. PATIENTS AND METHODS: Twelve patients with endoscopically proved gastric antral vascular ectasia were included. All patients received argon plasma coagulation with power of 40 W at a median interval of 4 weeks. The pre-treatment haemoglobin and transfusion requirements were compared with the post-treatment values. RESULTS: There was a sustained increase in mean haemoglobin levels post-treatment. The mean haemoglobin levels pre- and post-treatment were 8.13 +/- 0.70 and 12.2 +/- 0.32 g/dl, respectively (P = 0.008). All patients were anaemic and 58.3% of the patients were transfusion dependent. The mean number of units of blood transfusion in the period 6 months prior to treatment was 11.3 +/- 5.68. Following argon plasma coagulation, the number of transfusions decreased significantly to 1.1 +/- 0.57 units (P = 0.018). No significant procedure-related complications were identified. CONCLUSION: Argon plasma coagulation is a safe and effective alternative to the currently available endoscopic modalities of treatment for gastric antral vascular ectasia.

Review article: current therapeutic options for gastric antral vascular ectasia.

Gastric antral vascular ectasia, or 'watermelon stomach', is a rare but important cause of gastrointestinal blood loss and anaemia, which has characteristic endoscopic and histological features. The pathogenesis of this condition remains unclear; however, many associated disorders have been documented. Various medical, surgical and endoscopic therapeutic modalities have been attempted with variable success. Leading contenders for the therapeutic modality of choice include hormonal therapy, endoscopic Nd:YAG laser and argon plasma coagulation. Randomized controlled trials to identify the ideal treatment method are lacking at present.

New normalization methods for cDNA microarray data.

MOTIVATION: The focus of this paper is on two new normalization methods for cDNA microarrays. After the image analysis has been performed on a microarray and before differentially expressed genes can be detected, some form of normalization must be applied to the microarrays. Normalization removes biases towards one or other of the fluorescent dyes used to label each mRNA sample allowing for proper evaluation of differential gene expression. RESULTS: The two normalization methods that we present here build on previously described non-linear normalization techniques. We extend these techniques by firstly introducing a normalization method that deals with smooth spatial trends in intensity across microarrays, an important issue that must be dealt with. Secondly we deal with normalization of a new type of cDNA microarray experiment that is coming into prevalence, the small scale specialty or 'boutique' array, where large proportions of the genes on the microarrays are expected to be highly differentially expressed. AVAILABILITY: The normalization methods described in this paper are available via http://www.pi.csiro.au/gena/ in a software suite called tRMA: tools for R Microarray Analysis upon request of the authors. Images and data used in this paper are also available via the same link.

Differences in mandibular distraction osteogenesis after corticotomy and osteotomy.

Corticotomy or osteotomy was performed on opposing sides of the mandibles in 18 goats. A custom-made distractor was used to lengthen the mandible at a rate of 1 mm/day for 10 days (total 10 mm elongation). Six goats were sacrificed respectively at 2, 4 and 8 weeks after completion of distraction. The distracted calluses were harvested and processed for radiographic, histologic, and scanning electron microscopic evaluation as well as Ca/P ratio analysis. The regenerate bone in the corticotomy side showed more bone formation and earlier mineralization than in the osteotomy side. The results of this study suggest that preservation of intramedullary vessels is beneficial to bone regeneration following mandibular osteodistraction, and that performing corticotomy may be a simple but effective way to promote the maturity of the distracted callus and shorten the time for fixation.

Selective perceptual impairments after perirhinal cortex ablation.

It has been suggested that the primate perirhinal cortex contributes exclusively to memory. However, recent studies in macaque monkeys have implied that the perirhinal cortex may also contribute to object perception. To investigate whether the perirhinal cortex does contribute to perception, we devised several perceptual oddity tasks in which monkeys had to choose which stimulus of several presented concurrently on a touch screen was different. Macaques with bilateral perirhinal cortex ablations were selectively impaired relative to controls at perceptually discriminating the odd stimulus when the odd stimulus was a different object and when the discrimination could not be done on the basis of simple differences in features between the stimuli. They remained unimpaired relative to controls on discriminating the odd stimulus when the odd stimulus was a different color, a different shape, or a different size even when these discriminations were extremely difficult. They were also impaired on human and monkey face oddity tasks and oddity tasks with scenes containing objects. Therefore, we reject the notion that the macaque perirhinal cortex has a role exclusive to memory and conclude that the macaque perirhinal cortex does contribute to perception. We argue that the perirhinal cortex is neither specialized for perception nor memory processes alone, but rather, is specialized for processing stimuli that require processing at a more abstract level such as at the level of an object and that the perirhinal cortex contributes to both memory and perception of such stimuli.

Changes in the inferior alveolar nerve after mandibular lengthening with different rates of distraction.

PURPOSE: This study investigated the changes in the inferior alveolar nerve after mandibular lengthening with different rates of distraction. MATERIALS AND METHODS: Bilateral mandibular corticotomies were performed in 8 goats. The mandibles in 6 goats were lengthened 10 mm using a custom-made distractor with 2 different rates of distraction (1 mm/d [n = 3] and 2 mm/d [n = 3]); the other 2 nondistracted mandibles served as a control. The goats with distracted mandibles were killed at 2 weeks after completion of distraction. The inferior alveolar nerve specimens from all animals were harvested and processed for histologic and ultrastructural evaluation. RESULTS: The mandibles were lengthened successfully in the distracted animals. Morphologic changes in the inferior alveolar nerves were observed when compared with the nondistracted controls. Moreover, signs of nerve degeneration, such as demyelination, axonal swelling, axoplasmic darking, and decrease in the number of axons, were more extensive and prominent in those nerves distracted at a rate of 2 mm/d. CONCLUSIONS: Degenerative changes in the inferior alveolar nerve occur after mandibular lengthening by distraction osteogenesis. The distraction rate of 1 mm/d appears to be tolerable and safe for the inferior alveolar nerve, but rapid distraction may cause serious degeneration.

Cyclic tensile strain suppresses catabolic effects of interleukin-1beta in fibrochondrocytes from the temporomandibular joint.

OBJECTIVE: To discern the effects of continuous passive motion on inflamed temporomandibular joints (TMJ). METHODS: The effects of continuous passive motion on TMJ were simulated by exposing primary cultures of rabbit TMJ fibrochondrocyte monolayers to cyclic tensile strain (CTS) in the presence of recombinant human interleukin-1beta (rHuIL-1beta) in vitro. The messenger RNA (mRNA) induction of rHuIL-1beta response elements was examined by semiquantitative reverse transcriptase-polymerase chain reaction. The synthesis of nitric oxide was examined by Griess reaction, and the synthesis of prostaglandin E2 (PGE2) was examined by radioimmunoassay. The synthesis of proteins was examined by Western blot analysis of the cell extracts, and synthesis of proteoglycans via incorporation of 35S-sodium sulfate in the culture medium. RESULTS: Exposure of TMJ fibrochondrocytes to rHuIL-1beta resulted in the induction of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2), which were paralleled by NO and PGE2 production. Additionally, IL-1beta induced significant levels of collagenase (matrix metalloproteinase 1 [MMP-1]) within 4 hours, and this was sustained over a period of 48 hours. Concomitant application of CTS abrogated the catabolic effects of IL-1beta on TMJ chondrocytes by inhibiting iNOS, COX-2, and MMP-1 mRNA production and NO, PGE2, and MMP-1 synthesis. CTS also counteracted cartilage degradation by augmenting expression of mRNA for tissue inhibitor of metalloproteinases 2 that is inhibited by rHuIL-1beta. In parallel, CTS also counteracted rHuIL-1beta-induced suppression of proteoglycan synthesis. Nevertheless, the presence of an inflammatory signal was a prerequisite for the observed CTS actions, because fibrochondrocytes, when exposed to CTS alone, did not exhibit any of the effects described above. CONCLUSION: CTS acts as an effective antagonist of rHuIL-1beta by potentially diminishing its catabolic actions on TMJ fibrochondrocytes. Furthermore, CTS actions appear to involve disruption/regulation of signal transduction cascade of rHuIL-1beta upstream of mRNA transcription.

Analysis of cDNA microarray images.

Microarrays are part of a new class of biotechnologies that allow the monitoring of expression levels for thousands of genes simultaneously. Image analysis is an important aspect of microarray experiments, one that can have a potentially large impact on subsequent analyses, such as clustering or the identification of differentially expressed genes. This paper reviews a number of existing image analysis methods used on cDNA microarray data. In particular, it describes and discusses the different segmentation and background adjustment methods. It was found that in some cases background adjustment can substantially reduce the precision--that is, increase the variability of low-intensity spot values. In contrast, the choice of segmentation procedure seems to have a smaller impact.

Interaction of strain and interleukin-1 in articular cartilage: effects on proteoglycan synthesis in chondrocytes.

In temporomandibular joint disorders, the release of proinflammatory cytokines such as interleukin-1 (IL-1) initiates an inflammatory process disrupting cartilage homeostasis, ultimately leading to cartilage destruction. Additionally, mechanical stimuli affect articular chondrocyte metabolism. While articular chondrocytes generate nitric oxide (NO) in the presence of IL-1 proteoglycan synthesis is consecutively suppressed. The purpose of this study was to assess the effects of proinflammatory cytokines and mechanical strain in the form of cyclic tensile stretch on proteoglycan synthesis in chondrocytes, as compared to the NO competitive inhibitor L-N-monomethyl arginine (LMA), and to assess whether this effect is secondarily related to the activity of growth factors such as transforming growth factor beta (TGF-beta). Lapine articular chondrocytes were exposed to one of four different treatment regimens: no cyclic tensile stretch, IL-1, cyclic tensile stretch, or IL-1 plus cyclic tensile stretch. NO production was determined as medium nitrite accumulation. TGF-beta-bioactivity in chondrocyte conditioned medium was measured with the mink-lung epithelial cell bioassay. Proteoglycan synthesis was measured as the incorporation of 35-[S]-sodium sulfate into macromolecules separated from unincorporated label by gel filtration on PD-10 columns. In resting chondrocyte cultures, only baseline levels of NO were measured and the application of stretch for 24 h did not affect NO production. Addition of IL-1 provoked a large increase in NO synthesis which was abrogated in the presence of LMA. Application of stretch decreased the IL-1 induced NO synthesis, but did not modify the effect of LMA (being a competitive inhibitor of the inducible NO synthase) inhibiting IL-1 induced NO production. Glucosaminoglycan production was noted as proteoglycan synthesis showing almost no effect of cyclic stretch alone in comparison to the control condition, which correlates with the missing NO production in control and stretch conditions. Addition of IL-1 strongly inhibited proteoglycan synthesis, which was partly restored in the presence of LMA. However, cyclic stretch acted as a stronger restorer of proteoglycan synthesis in IL-1 treated conditions in the absence, and even more in the presence, of LMA. It was concluded that motion in the form of cyclic tensile stretch is a remarkable anti-inflammatory stimulus reversing the IL-1 induced suppression of proteoglycan synthesis in chondrocytes. These findings have therapeutic implications for the treatment of temporomandibular joint disorders, supporting early onset of postoperative and post-traumatic continuous passive motion therapy.

A new source of drug-induced acute pancreatitis: codeine.

A variety of drugs have been reported to cause acute pancreatitis during the past 40 years. We report the first series of four cases of acute pancreatitis related to codeine ingestion. Four patients (three female, mean age 50.2 yr) presented with clinical, biochemical, and radiological evidence of acute pancreatitis. All four had ingested a therapeutic dose of codeine 1-3 h before the onset of abdominal symptoms. Unintentional rechallenge occurred in three cases and was followed by recurrence of acute pancreatitis in all three. All patients made a full recovery. All four patients had had a previous cholecystectomy. The likely underlying pathophysiological mechanism is codeine-induced spasm of the sphincter of Oddi combined with sphincter of Oddi dysfunction related to a previous cholecystectomy. Codeine ingestion leads to acute pancreatitis in some individuals. Previous cholecystectomy seems to predispose to codeine-induced pancreatitis.

[Cytokine-induced nitric oxide production of joint cartilage cells in continuous passive movement. Anti-inflammatory effect of continuous passive movement on chondrocytes: in vitro study]. / Zytokininduzierte Stickstoffmonoxidproduktion von Gelenkknorpelzellen unter kontinuierlicher passiver Bewegung. Antientzündlicher Effekt von kontinuierlicher passiver Bewegung auf Chondrozyten: In-vitro-Nachweis.

PURPOSE: The temporomandibular joint is a place of motion where release of proinflammatory cytokines like interleukin-1 beta (IL-1 beta) induces cartilage destruction via production of nitric oxide (NO). The purpose of this study was to evaluate the effects of continuous passive motion in the form of cyclic stretch on the synthesis of inducible nitric oxide synthase (iNOS). METHODS: Articular chondrocytes were harvested from rabbit cartilage slices and cultured in F12 medium supplemented with 10% fetal calf serum. Subsequently, cells (10(5)/ml per well) were transferred to Flexercell plates, grown to 90% confluency, and subjected to one of the following regimens: (1) no mechanical strain (10 Hz, 20% elongation rate); (2) rhIL-1 beta (1 ng/ml); (3) mechanical strain; (4) mechanical strain and rhIL-1 beta. The cells were exposed to cyclic stretch for 24 h. Thereafter, cells were trypsinized and centrifuged on microscope slides in a cytospin centrifuge. The presence of iNOS was determined by indirect immunoperoxidase staining using polyclonal rabbit anti-iNOS as primary antibodies, and goat anti-rabbit IgG conjugated with horseradish peroxidase (HRP) as secondary antibodies. Diaminobenzidine was used as substrate for HRP. Total NO production was spectrophotometrically measured in the supernatants of cultures using Greiss reaction. All experiments were done in triplicates and the statistical significance was analyzed by Student's t-test. RESULTS: Cells with treatment 1 and 3 did not exhibit presence of iNOS. IL-1 beta-treated cells in group 2 exhibited intense peroxidase staining. Cells in group 4 exposed to IL-1 and mechanical stress showed staining with considerably less intensity. Control cells treated with normal rabbit IgG as a primary antibody did not exhibit peroxidase staining. The data were further confirmed by measurement of statistically significant differences of NO levels in supernatants assessed in the four different treatment groups. CONCLUSION: Our findings suggest that continuous passive motion exerts anti-inflammatory effects on chondrocytes, downregulating the quantity of iNOS-positive cells. Since NO acts as an intracellular, transcellular, and cytotoxic molecule, it has a basic importance for proper post-traumatic and postoperative TMJ function as well as the course and therapy of inflammatory TMJ diseases.

Growth factor and cytokine gene expression in mechanically strained human osteoblast-like cells: implications for distraction osteogenesis.

OBJECTIVE: An understanding of bone cellular biology is a basic necessity to understanding events such as distraction osteogenesis. The goal of this study was to determine the effect of continuous cyclic mechanical stretch as a fundamental event in distraction osteogenesis on the expression of 3 bone growth factors, transforming growth factor-beta 1 (TGF-beta1), insulin-like growth factor 1 (IGF-1), basic fibroblast growth factor (bFGF) and 2 cytokines, interleukin (IL)-1 (IL-1) and 6 (IL-6) in human osteoblast-like cells. MATERIAL AND METHODS: A human osteoblast-like cell line, SaOS-2, capable of forming a ground substance and mineralizing it, was maintained. Cells were transferred to 6-well plates with flexible silicon bottoms grown to confluence and either subjected to tensile stretch for different time intervals or used as the control group. RNA was isolated to conduct Northern blot analysis for the expression of 3 bone growth factors, transforming TGF-beta1, IGF-1, bFGF, and 2 cytokines, IL-1 and IL-6. RESULTS: After 8 hours, mRNA for TGF-beta1 and IGF-1 increased in the experimental group, whereas bFGF decreased but cytokines IL-1 and IL-6 were not affected. At 16 hours, TGF-beta1, IGF-1, and bFGF showed increased levels of mRNA; IL-6 showed a slight increase. After 24 hours, TGF-beta1, IGF-1, bFGF, and IL-6 had increased mRNA levels. IL-1beta did never show significant alterations in mRNA production as compared with the control. CONCLUSION: Tensile stretch on osteoblast-like cells alter local regulation of bone formation, increasing the expression of bone growth factors, whereas catabolic cytokines are unaffected. These findings suggest a direct effect of mechanical strain on osteoblasts and may be the driving factors of bone growth during distraction.