Connective tissue growth factor promotes fibrosis downstream of TGFbeta and IL-6 in chronic cardiac allograft rejection.

Cardiac transplantation is an effective treatment for multiple types of heart failure refractive to therapy. Although immunosuppressive therapeutics have increased survival rates within the first year posttransplant, chronic rejection (CR) remains a significant barrier to long-term graft survival. Indicators of CR include patchy interstitial fibrosis, vascular occlusion and progressive loss of graft function. Multiple factors have been implicated in the onset and progression of CR, including TGFbeta, IL-6 and connective tissue growth factor (CTGF). While associated with CR, the role of CTGF in CR and the factors necessary for CTGF induction in vivo are not understood. To this end, we utilized forced expression and neutralizing antibody approaches. Transduction of allografts with CTGF significantly increased fibrotic tissue development, though not to levels observed with TGFbeta transduction. Further, intragraft CTGF expression was inhibited by IL-6 neutralization whereas TGFbeta expression remained unchanged, indicating that IL-6 effects may potentiate TGFbeta-mediated induction of CTGF. Finally, neutralizing CTGF significantly reduced graft fibrosis without reducing TGFbeta and IL-6 expression levels. These findings indicate that CTGF functions as a downstream mediator of fibrosis in CR, and that CTGF neutralization may ameliorate fibrosis and hypertrophy associated with CR.

Validating and corroborating the deposition of two annual growth zones in asteriscus otoliths of common carp Cyprinus carpio from South Africa's largest impoundment.

A total of 816 common carp Cyprinus carpio asteriscus otolith pairs were collected from Lake Gariep, South Africa. Otoliths were interpreted whole, submerged in methyl salicylate and viewed under transmitted light. The precision of growth zone counts of the primary reader was estimated at 5·54 and 7·03% using the average per cent error method and the coefficient of variation, respectively. Age-bias plots indicated no systematic bias between the primary reader and the three secondary readers for up to nine growth zones (95% of the sample). Growth zone deposition rate was validated using a mark-recapture experiment of chemically tagged C. carpio (n = 21) conducted in a large earthen pond under ambient conditions in the vicinity of Lake Gariep. The validation results were corroborated for the wild population by edge analysis and a length-based age-structured model. All three methods suggest that growth zone formation occurred biannually, exemplifying the importance of age validation as a prerequisite for understanding the life history of C. carpio.

Critical role for IL-6 in hypertrophy and fibrosis in chronic cardiac allograft rejection.

Chronic cardiac allograft rejection is the major barrier to long term graft survival. There is currently no effective treatment for chronic rejection except re-transplantation. Though neointimal development, fibrosis, and progressive deterioration of graft function are hallmarks of chronic rejection, the immunologic mechanisms driving this process are poorly understood. These experiments tested a functional role for IL-6 in chronic rejection by utilizing serial echocardiography to assess the progression of chronic rejection in vascularized mouse cardiac allografts. Cardiac allografts in mice transiently depleted of CD4+ cells that develop chronic rejection were compared with those receiving anti-CD40L therapy that do not develop chronic rejection. Echocardiography revealed the development of hypertrophy in grafts undergoing chronic rejection. Histologic analysis confirmed hypertrophy that coincided with graft fibrosis and elevated intragraft expression of IL-6. To elucidate the role of IL-6 in chronic rejection, cardiac allograft recipients depleted of CD4+ cells were treated with neutralizing anti-IL-6 mAb. IL-6 neutralization ameliorated cardiomyocyte hypertrophy, graft fibrosis, and prevented deterioration of graft contractility associated with chronic rejection. These observations reveal a new paradigm in which IL-6 drives development of pathologic hypertrophy and fibrosis in chronic cardiac allograft rejection and suggest that IL-6 could be a therapeutic target to prevent this disease.

Healthcare-associated rotavirus gastroenteritis in a large paediatric hospital in the UK.

Rotavirus is a major cause of acute, community-acquired gastroenteritis in infants and young children but its importance in healthcare-associated paediatric gastroenteritis is much less well understood. A prospective study was undertaken at the Royal Liverpool Children's NHS Trust, Liverpool, UK. We enrolled 243 children with acute gastroenteritis (AGE) who were hospitalised between January and May 2006. Rotavirus was detected in faecal specimens by enzyme immunoassay. This virus was responsible for 17/91 cases (19%) of healthcare-associated AGE and 54/152 cases (36%) of community-acquired AGE. Sixteen children with healthcare-associated acute rotavirus gastroenteritis required source isolation and eight received intravenous rehydration. We conclude that rotavirus is an important cause of healthcare-associated AGE in a large paediatric hospital. Rotavirus vaccines represent a public health tool that could prevent the majority of symptomatic rotavirus infections within this healthcare setting.

Pseudopterygium arising in a patient with multiple keloids.

We describe an unusual case of mechanical restriction of the globe caused by pseudopterygium formation at the site of a minor ocular injury, which led to a possible link between Ebstein's anomaly and keloids.

The direct brow lift: efficacy, complications, and patient satisfaction.

BACKGROUND/AIMS: The direct brow lift operation can be used to treat brow ptosis arising from either involutional changes or facial nerve palsy. The authors reviewed their experience with this operation to establish its efficacy and complication rate in the light of concerns over poor scar cosmesis and forehead paraesthesiae in the postoperative period. METHODS: A retrospective review of patients undergoing direct brow lifting from 1989 to 2002 was conducted, and information gained on patient satisfaction by questionnaire. RESULTS: The direct brow lift operation was found to give a predictable outcome, with high levels of patient satisfaction. With careful wound closure, postoperative scars are rarely cosmetically unacceptable to the patient. Paraesthesiae are a common but well tolerated sequelae. CONCLUSIONS: The direct brow lift was found to be a reliable method for treating brow ptosis arising through involutional change or facial nerve palsy in both men and women. The postoperative scars may be more evident in younger patients so the authors reserve this technique for "rehabilitative" rather than cosmetic brow lifts in patients of middle age and beyond.

Case report: the pulmonary lesions of chickenpox pneumonia--revisited.

Infection with the varicella zoster virus can include pulmonary complications. These may cause such minor symptomatology as to go unrecognized and unimaged. We describe a case of active chickenpox pneumonia that mimicked pulmonary metastases and was detected incidentally by means of computed tomography in a man with a past history of testicular teratoma.

Ultrasound examination of the irritable hip.

We made a prospective study of 111 children with acute hip pain to assess whether ultrasound can replace traditional radiography. An effusion was diagnosed in 71% by ultrasound but in only 15% by radiography. This effusion persisted for a mean of nine days; symptoms lasted for five days. Two patients found to have Perthes' disease had longer-lasting effusion and symptoms. Patients without an effusion had no obvious cause for their pain, so the pressure of an effusion from a transient synovitis does not account for all patients with irritable hips. Patients with an effusion persisting for over 24 days (the mean + 2 s.d. of our series) had more symptoms, a significantly larger effusion and greater limitation of movement. They may be more at risk for avascular necrosis. We found that radiographic examination influenced the immediate management of only two patients, those with Perthes' disease. We therefore propose a protocol of management for irritable hip, using ultrasonography at the first presentation of certain categories of patients. This would reduce the number of early radiographs by 75%.

Correction of metabolic acidosis in diarrheal calves by oral administration of electrolyte solutions with or without bicarbonate.

Acid-base balance was evaluated in calves with experimentally induced viral diarrhea. When blood pH decreased to less than 7.200, calves were assigned to treatment groups and fed milk replacer, electrolyte solution without bicarbonate, or electrolyte solution containing bicarbonate. Calves in the electrolyte treatment groups had lower mortality (P less than 0.05), were better hydrated (P less than 0.05), and were less acidotic (P less than 0.05) than calves fed milk replacer. Bicarbonate-containing electrolyte solution restored acid-base balance (P less than 0.05) and corrected depression better (P less than 0.05) than electrolyte solution that did not contain bicarbonate. Both electrolyte solutions were equally good at correcting dehydration.

On the life history of the lesser gurnard (Scorpaeniformes: Triglidae) inhabiting the Agulhas Bank, South Africa.

Growth analysis based on sectioned sagittal otoliths revealed the lesser gurnard Chelidonichthys queketti on the Agulhas Bank to be relatively fast growing and long lived, with ages of up to 7 years being recorded. Total length at age (mm) was described best by the specialized von Bertalanffy growth model as LT =306·1 (1 - e0·53(t+0·18) ). First approximations of total, natural and fishing mortality rates were determined at 0·73, 0·38 and 0·35 year-1 respectively. The adult population was male dominated with a sex ratio of 1 female: 1·2 males with the mean size of males and females being similar. The lesser gurnard is an iteroparous species with females maturing by the end of the first year of life (195 mm LT ), thereafter spawning throughout the year with reproductive activity peaking over spring and late summer. The lesser gurnard appears to exhibit similar life-history patterns to other triglid species in that it can be classified as a generalist. Generalistic life-history characteristics such as a fast growth rate, early sexual maturity at a relatively large size, a non-seasonal spawning pattern, feeding on a variety of prey organisms and the ability to inhabit various substrata could all contribute to it maintaining a high biomass on the Agulhas Bank.

Salmon poisoning disease in dogs on southern vancouver island.

Salmon poisoning disease in dogs has previously been reported in North America only along the western coast of the U.S.A. This paper presents the findings from eight affected dogs recently diagnosed on Vancouver Island, Canada. The clinical signs shown by these dogs were lethargy, anorexia, pyrexia and lymph node enlargement. The causative agent, Neorickettsia helminthoeca was observed in macrophages obtained from lymph node aspirates. This organism is transmitted to dogs in cysts of the fluke Nanophyetus salmincola salmincola within the tissues of the salmon or trout. The presence of fluke eggs in the feces of dogs showing typical signs is very suggestive of a diagnosis of salmon poisoning disease. Appropriate treatment, including chloramphenicol or oxytetracycline and fluid therapy, resulted in recovery. Prevention of salmon poisoning disease in endemic areas can be achieved by advising owners against allowing their dogs to eat raw salmon or trout. We suggest, based on the diagnoses made in these eight dogs, that Vancouver Island now be considered an endemic area for salmon poisoning disease.

Ultra scale-down studies of the effect of flow and impact conditions during E. coli cell processing.

The ability to recover cells from a fermentation broth in an intact form can be an important criterion for determining the overall performance of a recovery and purification sequence. Disruption of the cells can lead to undesired contamination of an extracellular product with intracellular components and vice versa loss of intracellular products may occur. In particular, the value of directed location of a product in the periplasmic space of say Escherichia coli (E. coli) would be diminished by such premature non-selective cell disruption. Several options exist for cell recovery/removal; namely centrifugation, in batch or continuous configuration, filtration or membrane operations, and in selected cases expanded beds. The choice of operation is dependant on many variables including the impact on the overall process sequence. In all cases, the cells are exposed to shear stresses of varying levels and times and additionally such environments exist in ancillary operations such as pumping, pipe flow, and control valves. In this study, a small-scale device has been designed to expose cells to controlled levels of shear, time and impact in a way that seeks to mimic those effects that may occur during full-scale processes. The extent of cell breakage was found to be proportional to shear stress. An additional level of breakage occurred due to the jet impacting on the collecting surface. Here it was possible to correlate the additional breakage with the impact velocity, which is a function of the distance that the jet travels before meeting the collection surface and the initial jet velocity.

Fatal pulmonary embolism due to osteogenic sarcoma in a child.

A case of fatal pulmonary embolus in a 9-year-old child is presented, in which there is an unusual underlying cause. Whilst pulmonary embolism is rare in childhood, it must be considered when the clinical and radiological circumstances are appropriate. Extensive thrombosis of the lower limbs can occur without clinical signs and the value of venography is emphasised.

Modeling the performance of pilot-scale countercurrent chromatography: scale-up predictions and experimental verification of erythromycin separation.

Biosynthesis of polyketide antibiotics, such as erythromycin A (EA), can result in the formation of analogues of the main compound that are chemically and structurally extremely similar. The large-scale purification of these antibiotics by conventional high-performance liquid chromatography (HPLC) can be prohibitively expensive due to the large volume of both solvent and adsorbent required. This study examines the feasibility of using a novel pilot-scale countercurrent chromatography (CCC) machine as an alternative to HPLC. CCC is a low-pressure (typically <4000 kN m(-2)) liquid-liquid chromatographic technique that allows the separation of solutes on the basis of their partitioning between two immiscible liquid phases. The effects of mobile phase flow rate, column rotational speed, and sample injection volume on the attainable yield and purity of EA were investigated. Our results show that, at a mobile phase flow rate of 40 mL min(-1), a rotational speed of 1200 rpm, and an injection volume of 100 mL (10 g total erythromycin), EA could be satisfactorily fractionated with a purity of approximately 92% (w/w) and a recovery yield of approximately 100% (w/w). The total solute throughput was estimated to be 0.41 kg day(-1). More importantly, we demonstrated simple and predictive linear scale-up of the CCC separation based on data obtained from a single laboratory-scale CCC chromatogram, and verified this experimentally. The retention time and peak width of the target compound at the pilot scale could be predicted to within 4% for operation at a range of mobile-phase flow rates and injection volumes. This predictable nature of CCC separations, unlike HPLC methods, can greatly reduce process development times and enable a complete process-scale operating scenario to be planned.

Antibiotic purification from fermentation broths by counter-current chromatography: analysis of product purity and yield trade-offs.

Counter-current chromatography (CCC) is a low pressure, liquid-liquid chromatographic technique which has proven to be a powerful purification tool for the high-resolution fractionation of a variety of active pharmaceutical compounds. The successful integration of CCC into either existing or new manufacturing processes requires the predictable purification of target compounds from crude, fermentation-derived, feed streams. This work examines the feasibility of CCC for the purification of fermentation-derived erythromycin A (EA) from its structurally and chemically similar analogues. At the laboratory scale, the effect of feed pre-treatment using either clarified, forward extracted (butyl acetate) or back extracted broth on EA separation was investigated. This defined the degree of impurity removal required, i.e. back extracted broth, to ensure a reproducible elution profile of EA during CCC. Optimisation and scale-up of the separation studied the effects of mobile phase flow (2-40 ml.min(-1)) and solute loading (0.1-10 g) on the attainable EA purity and yield. The results in all cases demonstrated a high attainable EA purity (>97% w/w) with throughputs up to 0.33 kg.day(-1). Secondly, a predictive scale-up model was applied demonstrating, that from knowledge of the solute distribution ratio of EA (K(EA)) at the laboratory scale, the EA elution time at the pilot scale could be predicted to within 3-10%, depending upon the solute injection volume. In addition, this study has evaluated a "fractionation diagram" approach to visually determine the effects of key operational variables on separation performance. This resulted in accurate fraction cut-point determination for a required degree of product purity and yield. Overall, the results show CCC to be a predictable and scaleable separation technique capable of handling real feed streams.

Ultrasound diagnosis of splenic injuries in children--the importance of free peritoneal fluid.

Three cases of injury to the spleen in children are presented. The value of ultrasound in the diagnosis and monitoring of treatment of such splenic injuries is described, and the importance of free peritoneal fluid is stressed. Since splenectomy disturbs the immune mechanisms of the body, treatment should, if possible, be medical.

Waiting times and patient satisfaction in the accident and emergency department.

A survey of the waiting times and patients' opinions of these times was undertaken in a busy district general hospital A&E department. The various components of the overall waiting time are analysed and specific points of the patients' attendance, where waiting times were prolonged, are identified. Standards are derived which is hoped may result in 75% of patients being satisfied with the duration of their wait. The current levels of achievement are compared with these standards. Suggested and actual improvements to the department to improve our performance are described.