Cross-Linked Fiber Network Embedded in Elastic Matrix.

The mechanical behavior of a three-dimensional cross-linked fiber network embedded in matrix is studied in this work. The network is composed from linear elastic fibers which store energy only in the axial deformation mode, while the matrix is also isotropic and linear elastic. Such systems are encountered in a broad range of applications, from tissue to consumer products. As the matrix modulus increases, the network is constrained to deform more affinely. This leads to internal forces acting between the network and the matrix, which produce strong stress concentration at the network cross-links. This interaction increases the apparent modulus of the network and decreases the apparent modulus of the matrix. A model is developed to predict the effective modulus of the composite and its predictions are compared with numerical data for a variety of networks.

Evaluation of a decision support system to predict preoperative investigations.

BACKGROUND: We have developed the 'Optimising Surgical Care and Assessment Record' (OSCAR), a clinical decision support system, to help nurses in predicting necessary preoperative investigations before surgery. OSCAR applies the hospital's protocols, which are based on the National Institute for Health and Clinical Excellence guidelines, to the patient's medical history and surgical details before recommending required investigations. METHODS: We selected case notes of 50 patients randomly from the OSCAR system that were recorded between October 2006 and January 2007. To form a reference standard, these case histories were anonymized and then sent to 10 consultant anaesthetists across the country. They were asked to study the case history and choose which tests they would carry out and which they would not. Then we have evaluated OSCAR's ability to predict the necessary investigations and the nurses' judgement, in comparison with the reference standard. RESULTS: OSCAR's ability to identify which investigations should be carried out, that is, its sensitivity, was 91.5% and its ability to identify which investigations not to carry out, that is, its specificity, was 82.7%. OSCAR was consistent in predicting investigations for differing severities of surgery, for ASA grade and gender. We were unable to demonstrate any overall difference between OSCAR and the nurse's ability to predict preoperative investigations. When combining the nurse's predictions with OSCAR's recommendations, an even greater sensitivity of 98.2% could be achieved. CONCLUSIONS: OSCAR's prediction algorithm cannot replace the nurse's judgement, but it can be used as a supplementary decision aid to promote consistency and improve accuracy.

Rat supraspinatus tendon expresses cartilage markers with overuse.

The goals of this study were to investigate the response of the rat supraspinatus tendon to overuse at the molecular level using transcriptional profiling, and to identify potential markers of tendinopathy. Adult rats were subjected to an overuse protocol that consists of downhill running (10% grade) at 17 m/min for 1 h/day, 5 days/week, for a total of either 1, 2, or 4 weeks. Another group of rats served as nonrunning time 0 controls. Transcriptional profiling was performed on the supraspinatus and patellar tendons using an Affymetrix rat genome array. A gene was considered to be differentially expressed if the p value from an ANOVA test was less than 0.01 and the difference between runners and controls was at least twofold at any time point. The supraspinatus tendon had increased expression of well-known cartilage genes such as col2a1, aggrecan, and sox9. These genes were not regulated in the patellar tendon, an internal comparator. Few genes associated with inflammation, or angiogenesis, were differentially expressed, and no significant change in the regulation of matrix metalloproteinases was detected. The results of this study suggest that by expressing more cartilage genes, the tendon is converting toward a fibrocartilage phenotype as a result of the repetitive loading and repeated compression of the tendon as it passes through the acromial arch.

Large-scale purification of human islets utilizing discontinuous albumin gradient on IBM 2991 cell separator.

A new method is described for the large-scale purification of human pancreatic islets with a discontinuous gradient of bovine serum albumin formed on an IBM 2991 cell separator. Fifteen human pancreases were processed, and after density-gradient centrifugation, a mean of 2643 islets/ml pancreatic digest were recovered with a mean purity of 63% and contained in 430 microliter mean vol. Viability of gradient-isolated islets was compared with that of non-density-gradient islets (handpicked) and showed no difference in function. This technique allows isolation of intact, viable human islets of Langerhans of sufficient purity for potential human transplantation.

Successful reversal of diabetes in nude rats by transplantation of isolated adult human islets of Langerhans.

A method is described in which the viability of isolated adult human islets of Langerhans can be assessed in vivo. The Rowett nude rat, made diabetic with streptozocin (STZ), has been used as the islet recipient in these studies. Although these animals are athymic and are able to accept xenogeneic grafts for prolonged periods, they are very susceptible to dehydration and infection once made diabetic. Therefore, a considerably shortened diabetes induction period was used. The basis of the study was to prepare pure adult human pancreatic islets that were cultured for 48 h. Nude rats were given 80 mg/kg i.v. STZ during islet isolation and were transplanted with 800-1000 islets under the renal capsule at 48 h. To monitor islet function, animals were bled regularly for random blood glucose measurements and were given a glucose tolerance test at day 20. The kidney containing the graft was removed on day 21 to allow histological assessment of the graft and to confirm that glucose control was due to the transplanted islets and was not secondary to reversion of the animal's own islets. Seven rats were transplanted, and five were deemed to have received viable human islets. Two rats that received islets from the same donor did not reverse their diabetes and were found by histology to have vacuolated islet structures with scant insulin-staining tissue under the kidney capsule. This method allows a definitive judgment of the ability of isolated adult human islets to reverse diabetes.

Using topical negative pressure therapy to resolve wound failure following perineal resection.

Elective resection of the rectum and perineum in a patient with recurrent squamous cell carcinoma of the anus resulted in significant wound failure. Full healing was achieved with topical negative pressure, with no recurrence of the cancer.

Differential effects of beta-adrenergic agonists on insulin secretion from pancreatic islets isolated from rat and man.

The selective beta 2-adrenergic agonist clenbuterol was ineffective as a stimulus for insulin secretion when isolated rat pancreatic islets were incubated with glucose at concentrations between 4 and 20 mM. Inclusion of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine led to potentiation of glucose-induced insulin secretion, but did not facilitate stimulation by clenbuterol. Furthermore, maintenance of isolated rat islets for up to 3 days in tissue culture also failed to result in the appearance of a secretory response to beta-agonists. By contrast, clenbuterol induced a dose-dependent increase in insulin release from isolated human islets incubated with 20 mM glucose. Clenbuterol did not increase the basal rate of insulin secretion (4 mM glucose) in human islets. Under perifusion conditions, the secretory response of human islets to clenbuterol was rapid, of similar magnitude to that seen under static incubation conditions and could be sustained for at least 30 min. The increase in insulin secretion induced by clenbuterol was inhibited by propranolol, indicating that the response was mediated by activation of beta-receptors. In support of this, a similar enhancement of glucose-induced insulin secretion was elicited by a different beta 2-agonist, salbutamol, in human islets. The results indicate that the B cells of isolated rat islets are unresponsive to beta-agonists, whereas those of human islets are equipped with functional beta-receptors which can directly influence the rate of insulin secretion.

Bovine serum albumin density gradient isolation of rat pancreatic islets.

The use of a bovine serum albumin (BSA) density gradient for isolation of rat pancreatic Islets of Langerhans after collagenase digestion has been compared with the standard Ficoll separation technique. The criteria studied were islet yield (insulin extraction of the islet interfaces and pellet), purity of preparation (amylase content of the islet preparation), insulin release characteristics, and the result of isologous transplantation in diabetic rats. The islet interface of the BSA gradient contained 62.2% of the total insulin, whereas the corresponding interface of the Ficoll gradient contained only 36.6% (P less than 0.001). The amylase content of the Ficoll-separated islet preparation was 6133 U/L, as opposed to 1230 U/L with BSA (P less than 0.001). BSA-isolated islets gave similar insulin release characteristics to non-density-gradient-isolated islets, whereas Ficoll-separated islets showed suboptimal insulin release. Single-donor-single-recipient transplantation was successfully performed with BSA-isolated islets whereas multiple donors were required with Ficoll-separated islets. Thus significantly improved results were found with the bovine serum albumin density gradient separation in all criteria and consequently the use of this gradient represents an advance in islet isolation techniques.

Gamma irradiation of isolated rat islets pretransplantation produces indefinite allograft survival in cyclosporine-treated recipients.

In this study we have examined the use of low-dose gamma-irradiation for the reduction of islet immunogenicity in the strong allogeneic combination of WAG rat islets transplanted into diabetic AUG recipients. First, we determined that gamma-irradiation reduced immunogenicity in vitro by use of a modified MLR with WAG islets as stimulators and AUG splenocytes as responders. We then determined the maximum dose of gamma-irradiation that could be used (250 rads) before islet function was affected. As 250 rads islet pretreatment alone was ineffective in prolonging allograft survival, we combined the pretreatment with a short course (days 0, 1, 2; 30 mg/kg) of cyclosporine. We found that CsA was only effective in significantly prolonging allograft survival when given subcutaneously in olive oil. The CsA treatment alone gave a significantly prolonged survival time for the islet allografts (median, 37 days vs. 6 days for controls), but when combined with the 250 rads islet pretreatment a synergistic effect was seen with 100% becoming long-term survivors (greater than 100 days). The long-term surviving AUG rats from both the CsA alone group and the CsA plus 250 rads pretreated islets group were challenged with WAG dendritic cells (DC). The islets from the 250 rads pretreated group were subsequently rejected (day 6) while the CsA alone group were not affected. The role of low dose gamma-irradiation when combined with CsA treatment of islet graft recipients in inducing specific unresponsiveness will be discussed.

A simple method for the release of islets by controlled collagenase digestion of the human pancreas.

A simple technique for the controlled collagenase digestion of the human pancreas is described. The pancreas is distended with collagenase, and a biopsy taken and divided into 5 pieces that are placed in Universals containing minimal essential medium and dithizone at 39 degrees C. The pancreas itself is incubated in MEM at 39 degrees C. Starting at 5 min and at intervals thereafter, a Universal is removed from the water bath, shaken for 30 sec, and the contents examined by microscopy. As soon as free cleaved islets are seen, the pancreas is placed into one compartment of a kidney-bowl divided in half by a 1-mm mesh. The pancreas is gently teased apart and fluid digest in the empty half of the bowl aspirated and passed through a 500-micron mesh into ice-cold MEM containing 20% newborn calf serum. This process is repeated until the digestion process has ceased. Using this technique on 20 consecutive pancreata, median wt. (range) 53.9 (45.2-72.9) g, we have counted 131,672 (43,516-400,000) islets in the digest, equivalent to 2394 (715-8000) islets/g pancreas. The volume of islet tissue in the digest was 299 (26-1341) mm3 equivalent to 5.81 (0.36-26.81) mm3/g pancreas. In conclusion, we have found this simple technique to be an effective method for the controlled collagenase digestion of the human pancreas.

HLA DR, DP, DQ induction in human islet beta cells by the cytokine combination IFN-gamma + TNF-alpha.

Human islet beta cells do not express HLA Class II normally, yet, in the diabetic pancreas, beta cells are selectively positive for Class II and this may facilitate their recognition by T cells. It has been demonstrated that human beta cells can be induced to express Class II when cultured with IFN-gamma + TNF-alpha or IFN-gamma + TNF-beta. To assess whether or not they can be induced to express the products of the Class II subregions, DR, DP and DQ, human islet cultures from 10 pancreas were supplemented with the combination of IFN-gamma + TNF-alpha using MoAbs specific for DR, DP and DQ products, and antibodies to insulin and glucagon. The combination IFN-gamma + TNF-alpha (100-1000 U/ml each) was able to induce the expression of the three subregions in both beta and alpha cells. The induction of subregion expression followed the hierarchy DR greater than DQ greater than or equal to DP. The capability of beta cells to express all three Class II subregions supports the possibility that these cells can present their self antigens to T cells.

Management of pelvic sepsis after Ivalon rectopexy.

The consequences of pelvic sepsis after Ivalon rectopexy are described in four patients. Despite clear evidence of pelvic infection, reoperation was delayed by ineffective conservative measures and morbidity thereby prolonged. In three, the causative organism was Staphylococcus aureus and it is suggested that prophylactic antimicrobial regimens for intestinal organisms alone may be inadequate.

Serum gastrin levels in the pre-operative localization and surgical treatment of gastrinomas.

The introduction of sensitive radioimmunoassays for gastrin has led to the earlier and more accurate diagnosis of gastrinomas and, as methods for tumour localization both pre- and intraoperatively have improved, the emphasis of surgery has changed from control of gastric acid secretion to tumour removal. We present three cases of sporadic gastrinoma who underwent exploratory laparotomy. In two cases, gastrinomas were discovered and excised resulting in cure for one patient. The third case underwent a negative laparotomy. The changes in serum gastrin levels taken during and immediately after surgery were related to the success or otherwise of tumour removal in each of the three cases. As in parathyroid surgery, with the development of rapid radioimmunoassays, the intraoperative measurement of declining serum gastrin levels will help in the early definition of surgical success supplementing frozen section and clinical judgement and improving patient management.

A randomized trial to compare single with multiple phenol injection treatment for haemorrhoids.

One hundred and twenty consecutive patients were entered into a randomized trial of single versus multiple phenol injection for the treatment of haemorrhoids. Follow-up at 3 and 12 months was available in 105 patients (56 in the single group and 49 in the multiple group). The results have shown that injection therapy, whether this be single or multiple, is an extremely effective form of therapy for patients with first or second degree haemorrhoids.

In vivo assessment of isolated pancreatic islet viability using the streptozotocin-induced diabetic nude rat.

In order to establish a model for the in vivo assessment of islet function we have used the Rowett nude rat with transplantation of allogeneic and xenogeneic (mouse) islets into the renal subcapsular space following a minimal period of diabetic induction. Thirty-one nude rats were given streptozotocin and 30 became diabetic with blood glucose levels of greater than 20 mmol/l at 48 h. Rat and mouse islets were prepared by intraductal collagenase and bovine serum albumin density gradient isolation. Eight rats received transplants of freshly prepared allogeneic islets and 8 rats received transplants of 48 h cultured allogeneic islets. Seven rats received transplants of 48 h cultured mouse islets. Diabetes was reversed in all animals and all remained normoglycaemic for 21 days. Graft removal by nephrectomy resulted in hyperglycaemia in 22 out of 23 animals. Histological examination of the grafts showed a band of endocrine tissue beneath the renal capsule which stained strongly positive for insulin and there was no evidence of lymphocytic infiltration/rejection. One rat remained normoglycaemic after graft removal, which may represent recovery of the animal's own islets from the streptozotocin-induced diabetes. Control rats remained diabetic until death. In conclusion, the athymic nude rat can be used for the assessment of allogeneic and xenogeneic islet function when a short (48 h) period of streptozotocin-induced diabetes is used. This model offers a potential method for assessing in vivo function of isolated human islets.

Mycotic aneurysm of the popliteal artery following right hemicolectomy.

A case of mycotic aneurysm of the popliteal artery, following right hemicolectomy for carcinoma, is described. This was treated by ligation and excision without bypass grafting. The clinical features are discussed and the literature reviewed.

Renal subcapsular islet transplantation in the rat: a comparison of three techniques.

The purpose of this study was to investigate whether the results of renal subcapsular islet transplantation in the rat could be improved by placing the islets in a blood or plasma clot. Islets were isolated and transplanted into diabetic recipients using a standardized technique and islet function post transplantation measured by the ability to restore normoglycemia and by intravenous glucose tolerance testing. It was found that 750 islets in minimal essential medium reversed diabetes (6/6 recipients) and produced a normal IVGTT whereas the success rate if the islets were placed in a blood or plasma clot was only 0/6 and 1/6 recipients respectively (p less than 0.008). We conclude that any possible benefit from trophic factors within blood or plasma is outweighed by the fact that islets in a blood or plasma clot are distanced from the blood supply on the kidney surface. This finding has important implications for human islet transplantation.