A dermal equivalent developed from fibroblast culture alone: Effect of EGF and insulin

We have recently developed a new dermal equivalent without exogenous materials by culturing dermal fibroblasts alone in serum-containing medium treated with several supplements. In this study, we investigated the effects of epidermal growth factor (EGF) and insulin on the formation of a dermal equivalent. After cultured dermal fibroblasts reached a confluence in serum-containing medium, they were treated with EGF or insulin. The combined effects of EGF and insulin were also studied. Macroscopically, in contrast to the culture without supplement, the addition of EGF or insulin produced a fibrous sheet. The combination of EGF and insulin showed a more marked effect than a single factor. Histologically, EGF or insulin alone induced a three-dimensional tissue containing several layers of fibroblasts. The combination of EGF and insulin produced a thicker tissue. It was composed of abundant extracellular matrix containing fibroblasts, suggesting a dermis-like tissue. It revealed collagen fibers by Masson-trichrome staining. Immunohistochemically, the components of dermal extracellular matrix such as type 1 collagen, elastin, and fibrillin-1 were diffusely expressed. Ultrastructurally, a large number of collagen fibrils with cross-striated patterns were found around the fibroblasts. These results showed that a dermal equivalent could be formed by culturing dermal fibroblasts alone postconfluently in serum-containing medium with EGF and insulin. They suggest that the two factors play an important role in the formation of a dermal equivalent.     

Bone Mineral Density Determinations by Dual-Energy x-ray Absorptiometry in the Management of Patients with Marfan Syndrome—Some Factors Which Affect the Measurement

Reduced bone mineral density (BMD) was sporadically reported in patients with Marfan syndrome. This may or may not place the Marfan patient at increased risk for bone fracture. In comparing the BMDs of our patients with those reported in the literature, it seemed that agreement between values, and hence the degree of osteoporosis or osteopenia reported, was dependent on the instrumentation used. The objective of this study was to statistically assess this impression. Bone mineral density measurements from our previously published study of 30 adults with Marfan syndrome performed on a Lunar DPXL machine were compared with studies published between 1993–2000 measured using either Lunar or Hologic bone densitometry instruments. The differences of our measurements compared with those made on other Lunar machines were not statistically significant, but did differ significantly with published results from Hologic machines (P < 0.001). Before progress can be made in the assessment of BMD and fracture risk in Marfan patients and in the evidence-based orthopedic management of these patients, standardization of instrumental bone density determinations will be required along with considerations of height, obesity, age, and sex.     

AlphaV beta3 (αvβ3) integrin inhibition reduces leukocyte–endothelium interaction in a pressure-induced reperfusion model

The leukocyte-endothelium interaction is known to contribute to reperfusion injury, which is considered to participate in the pathophysiology of pressure ulcers, and integrin alphaV beta3 (alphavbeta3) has been shown to mediate the processes of cellular adhesion in various types of cells. This study aims to clarify leukocyte behavior in our original microcirculatory pressure-induced reperfusion model, which can visualize the microcirculation in vivo. We also estimated the effect of alphavbeta3 integrin inhibition on the reduction of the leukocyte-endothelium interaction. Mice with dorsal skinfold chambers were divided into three groups: the baseline group (n=6), in which animals received no compression; the compression-reperfusion group (n=6), in which animals underwent 2-hour compression of the dorsal skin, followed by release, and the inhibitor-treated group (n=7), in which an alphavbeta3 inhibitor, CP4715, was administered in addition to the compression-release procedure. Staining with rhodamine 6G quantitatively visualized leukocyte behavior under the intravital fluorescent microscope. Compression-reperfusion induced a significant increase in rolling, sticking, and extravasation of the leukocytes. Treatment with the inhibitor strikingly reduced leukocyte sticking and extravasation. The present experiment has provided evidence that alphavbeta3 inhibition reduces leukocyte-endothelium interaction in our original pressure-induced reperfusion model.     

Doctors as managers: investors and reluctants in a dual role.

Government reform of the NHS in the UK has sought to increase the involvement of doctors (clinicians) in hospital management. Using frameworks from the psychological contract and organisational misbehaviour literatures, this paper examines the processes involved when clinicians assume management roles. This literature seeks to explain breaches to expectations regarding prior agreements with management and subsequent actions of 'getting even' as a result of breaches to the employment relationship. A qualitative methodology using interviews was undertaken, which identified two distinct groups of clinician-manager. Investors actively pursued a management opportunity as an alternative to clinical medicine, whilst reluctants tended to assume a management role to protect particular specialities from outside influence or from those they thought would be inappropriate clinician-managers. Investors and reluctants often had very little prior experience of management and managers and had problems reconciling their dual clinician-management role. Poor relationships with hospital managers who often had no understanding of their dual responsibilities led to tensions and conflict, which questions continued developments in this important area of UK health policy. Suggestions for improving this process are outlined.     

Influence of Chewing and Clenching on Salivary Cortisol Levels as an Indicator of Stress

PURPOSE: The purpose of this study was to investigate the effects of chewing and clenching on salivary cortisol levels as an indicator of stress. MATERIALS AND METHODS: Seventeen healthy dentulous subjects were given arithmetic exercises to perform within a 20-minute time limit in order to elicit stress (stress loading). In the first experiment (chewing), after stress loading, the subjects were asked to chew a paraffin wax while reading printed material (books, magazines, etc.) in silence for 10 minutes. The same procedure was then carried out again for control purposes, but this time the subjects were not required to chew wax. In the second experiment (light clenching), after stress loading, the subjects were required to carry out 5 seconds of light clenching followed by 5 seconds of rest repeatedly over a 3-minute period. The whole 3-minute process was repeated a total of three times. The control data for this second experiment consisted of measurements taken during the rest periods. Saliva specimens were collected in both experiments both before stress loading and after each procedure during 1-minute intervals to measure cortisol levels. RESULTS: In the chewing experiment, salivary cortisol levels were significantly reduced by chewing, compared with those in the controls (p < 0.05). This reduction in salivary cortisol was observed during chewing over a 10-minute period following stress loading. In the clenching experiment, salivary cortisol levels also showed a significant reduction during clenching, compared with those in the controls (p < 0.05). CONCLUSIONS: These results suggest that chewing and clenching promote relaxation in subjects under stress.     

The αvβ6 integrin plays a role in compromised epidermal wound healing

The alphavbeta6 integrin is an exclusively epithelial integrin that is highly expressed during fetal development. In adult tissue, alphavbeta6 integrin is expressed during inflammation, carcinogenesis, and in wound healing. We previously reported that alphavbeta6 integrin is highly expressed in poorly healing human wounds and its over-expression is associated with chronic wounds in a mouse model. The objective of this study was to investigate the role of alphavbeta6 integrin in compromised wound healing induced by hydrocortisone treatment or aging by using young and old mice deficient in or overexpressing the beta6 integrin subunit in the epidermis. Untreated aged beta6 integrin-deficient (beta6-/-) animals showed a significant delay in wound healing when compared to their age-matched controls or younger beta6-/- mice. The most significant delay was observed at the stages where granulation tissue deposition was occurring. Hydrocortisone treatment significantly delayed wound healing in wild-type and beta6 integrin-deficient mice in comparison with the untreated controls. However, hydrocortisone treatment in beta6 integrin overexpressing animals did not cause a significant delay in wound healing. The results of this study suggest that alphavbeta6 integrin plays an important role in wound healing in animals compromised by either age or stress mimicked by hydrocortisone.     

An Independent Evaluation of Four Quantitative Emergency Department Crowding Scales

Background Emergency department (ED) overcrowding has become a frequent topic of investigation. Despite a significant body of research, there is no standard definition or measurement of ED crowding. Four quantitative scales for ED crowding have been proposed in the literature: the Real‐time Emergency Analysis of Demand Indicators (READI), the Emergency Department Work Index (EDWIN), the National Emergency Department Overcrowding Study (NEDOCS) scale, and the Emergency Department Crowding Scale (EDCS). These four scales have yet to be independently evaluated and compared. Objectives The goals of this study were to formally compare four existing quantitative ED crowding scales by measuring their ability to detect instances of perceived ED crowding and to determine whether any of these scales provide a generalizable solution for measuring ED crowding. Methods Data were collected at two‐hour intervals over 135 consecutive sampling instances. Physician and nurse agreement was assessed using weighted κ statistics. The crowding scales were compared via correlation statistics and their ability to predict perceived instances of ED crowding. Sensitivity, specificity, and positive predictive values were calculated at site‐specific cut points and at the recommended thresholds. Results All four of the crowding scales were significantly correlated, but their predictive abilities varied widely. NEDOCS had the highest area under the receiver operating characteristic curve (AROC) (0.92), while EDCS had the lowest (0.64). The recommended thresholds for the crowding scales were rarely exceeded; therefore, the scales were adjusted to site‐specific cut points. At a site‐specific cut point of 37.19, NEDOCS had the highest sensitivity (0.81), specificity (0.87), and positive predictive value (0.62). Conclusions At the study site, the suggested thresholds of the published crowding scales did not agree with providers' perceptions of ED crowding. Even after adjusting the scales to site‐specific thresholds, a relatively low prevalence of ED crowding resulted in unacceptably low positive predictive values for each scale. These results indicate that these crowding scales lack scalability and do not perform as designed in EDs where crowding is not the norm. However, two of the crowding scales, EDWIN and NEDOCS, and one of the READI subscales, bed ratio, yielded good predictive power (AROC >0.80) of perceived ED crowding, suggesting that they could be used effectively after a period of site‐specific calibration at EDs where crowding is a frequent occurrence.     

Chronic lower extremity ischemia: a human model of ischemic tolerance.

BACKGROUND: Ischemic preconditioning (IPC) has been found in animals to have a protective effect against future ischemic injury to muscle tissue. Such injury is unavoidable during some surgical procedures. To determine whether chronic ischemia in the lower extremities would imitate IPC and reduce ischemic injury during vascular surgery, we designed a controlled clinical study. PATIENTS AND METHODS: Two groups of patients at a university-affiliated medical centre with chronic lower-extremity ischemia served as models of IPC: 6 patients awaiting femoral distal bypass (FDB) and 4 scheduled for aortobifemoral (ABF) bypass grafting for aortoiliac occlusive disease. Seven patients undergoing elective open repair of an infrarenal abdominal aortic aneurysm (AAA) were chosen as non-IPC controls. Three hematologic indicators of skeletal-muscle injury, lactate dehydrogenase (LDH), creatine kinase (CK) and myoglobin, were measured before placement of the proximal clamp, during surgical ischemia, immediately upon reperfusion, 15 minutes after and 1 hour after reperfusion, and during the first, second and third postoperative days. RESULTS: Baseline markers of skeletal-muscle injury were similar in all groups. In postreperfusion samples, concentrations of muscle-injury markers were significantly lower in the 2 PC groups than in the control group. For example, at day 2, LDH levels were increased by about 30% over baseline measures in the elective AAA (control) group, whereas levels in the FDB and ABF groups remained statistically unchanged from baseline. Myoglobin in controls had increased by 977%, but only by 160% in the FDB and 528% in the ABF groups. CK levels, in a similar trend, were 1432% higher in the control group and only 111% (FDB) and 1029% (ABF) in the study groups. Taken together, these data represent a significant level of protection. CONCLUSIONS: Patients with chronic lower-extremity ischemia suffered less severe ischemic injury after a period of acute ischemia than those with acute ischemia alone. Ischemic preconditioning is one proposed mechanism to help explain this protective effect.