A simple method for coating native polysaccharides onto nitrocellulose.

A method for coating native, non-derivatized, polysaccharide (PS) onto nitrocellulose (NC) for identifying PS-specific antibodies has been developed. The new feature of this method is that PS molecules are vacuum filtered onto NC in their native state by devices that can accommodate NC of different sizes and shapes. PS-coated NC disks were used to localize antibody secreting hybridoma cells cultured on filter paper disks. These were analyzed by blotting with size-matched PS-coated NC disks and specific antibodies secreted by individual colonies were detected by enzyme-linked immunoblot. In another application of this method, immune sera were separated by isoelectric focusing and the gels were blotted with PS-coated NC sheets. The spectrotype and isotype of antibodies that bound to the NC were examined using isotype specific enzyme-linked antibody. These immunoblots showed high resolution and specificity. The advantages of this method are that the PS used for coating does not need to be derivatized in order to bind the NC, and that smaller quantities of PS may be utilized by this coating method when compared to other techniques. This provides a useful tool to ask many questions regarding the immune response to PS.     

Job ending among youth and adults with severe mental illness

This study examined job leaving over a 36-month period among 326 persons (74 youth and 252 adults) with severe mental illness who were participating in an urban vocational rehabilitation program. Data from 627 job endings indicated that younger clients displayed job-ending patterns that were different in some aspects from such patterns for nondisabled youth (such as displaying a lower average job tenure) yet similar to job-ending patterns for nonhandicapped youth in other ways (such as displaying a high frequency of job changing). Youth and their adult counterparts with mental illness displayed similar tenure on agency-sponsored placements but significantly different tenure on independent jobs. While adults held their independent jobs for an average of seven months, youth averaged only three months at competitive employment. Youth also were significantly more likely than adults to be fired from both placements and independent jobs. These findings and others are discussed in terms of their implications for development of public policy and service delivery models for providing on-going job support to youth and adults with severe psychiatric disabilities.     

Inhibin and age in men

OBJECTIVE: Normal elderly men are reported to have decreased testicular function despite elevated gonadotrophin levels. We wished therefore to determine if changes in testicular function occur over the age range 19-60 years. DESIGN: Single fasting blood samples were obtained between 0800 and 0900 h. PATIENTS: Working men in a large industrial company between the ages of 19 and 60 years participated in the study. MEASUREMENTS: FSH, serum immunoreactive inhibin and total testosterone were measured, the latter two as measurements of Sertoli and Leydig cell function respectively. RESULTS: The mean baseline serum immunoreactive inhibin level was significantly lower in men from the older age groups, 31-40 years (479 U/l), 41-50 years (439 U/l) and 51-60 years (415 U/l) than in men from the youngest age group, 21-30 years (613 U/l) while serum FSH was higher in men from the older age groups, 41-50 years (3.7 IU/l) and 51-60 years (6.1 IU/l) than in men from the youngest age group, 21-30 years (2.6 IU/l). There appears to be a change in both FSH and inhibin production, consistent with a primary decline in testicular function. There was no significant difference in testosterone levels between the older age group, age 51-60 years and the younger age group, age 21-30 years. However, testosterone levels were significantly lower in the 41-50 year age group, when compared with the 21-30 year, this significance levelling out at about age 45 years. CONCLUSION: The data are consistent with the hypothesis that immunoreactive inhibin reflects inhibin bioactivity, and that inhibin plays a role in the feedback control of FSH secretion in men.     

TGFβ-1 regulation of VEGF production by breast cancer cells

Background: Angiogenesis is essential for tumor growth and metastasis. Vascular endothelial growth factor (VEGF) is the most potent angiogenic factor identified to date. TGFβ-1 acts as an indirect angiogenic agent. Methods: VEGF and TGFβ-1 were measured in the serum of breast cancer patients and agematched controls and in tumor tissue of cancer patients by ELISA. VEGF protein and mRNA expression by breast tumor cell lines were examined, and the effect of TGFβ-1 on VEGF production in these cells was assessed. Results: VEGF levels were significantly higher (P=.03) in the serum of patients with breast cancer compared to age-matched controls. A positive correlation was found between serum (r=0.539) and tumor tissue (r=0.688) levels of VEGF and TGFβ-1. Metastatic MDA-MB-231 breast cancer cells produce more VEGF than do the primary BT474 cells. TGFβ-1 significantly (P<.05) increased production of VEGF. Conclusions: Breast cancer cells constitutively produce VEGF protein and mRNA. There is a relationship between VEGF and TGFβ-1 levels in breast cancer patients, and TGFβ-1 regulates VEGF expression by breast cancer cells. Presented at the 50th Annual Cancer Symposium of the Society of Surgical Oncology, Chicago, Illinois, March 20–23, 1997.     

Mechanics of the anterior drawer and talar tilt tests: A cadaveric study of lateral ligament injuries of the ankle

We analyzed the changes in lateral ligament forces during anterior drawer and talar tilt testing and examined ankle joint motion during testing, following an isolated lesion of the anterior talofibular ligament (ATFL) or a combined lesion of the ATFL and calcaneofibular ligament (CFL). 8 cadaver specimens were held in a specially designed testing apparatus in which the ankle position (dorsiflexion-plantarflexion and supination-pronation) could be varied in a controlled manner. Ligament forces were measured with buckle transducers, and joint motion was measured with an instrumented spatial linkage. An anterior drawer test was performed using an 80 N anterior translating force, and a talar tilt test was performed using a 5.7 Nm supination torque with intact ligaments, after sectioning of the ATFL, and again after sectioning of the CFL. The tests were repeated at 10° dorsiflexion, neutral, and 10° and 20° plantarflexion. In the intact ankle, the largest increases in ATFL force were observed during testing in plantarflexion, whereas the largest increases in CFL force were observed in dorsiflexion. Isolated ATFL injury caused only small laxity changes, but a pronounced increase in laxity was observed after a combined CFL and ATFL injury.     

Imaging strategies in children with suspected appendicitis

Appendicitis is the commonest acute surgical emergency of childhood, and occurs in approximately 2–4 per 1000 infants. It is usually seen in infants older than 5 years but can occur at any age. Atypical clinical findings are seen in 30–50% of children, especially the younger ones, and often leads to a delayed diagnosis. Preoperative clinical assessment alone has yielded an accuracy ranging between 70 and 90% but is associated with a normal appendectomy rate of 13–25%. Preoperative imaging using the graded compression US technique and/or different helical CT techniques has been able to reduce this rate to 3–7% without an increase in perforation rate. An extensive review of the literature revealed several papers examining the accuracy of different imaging modalities and strategies of acute appendicitis in children. The reported sensitivity of US varied between 87 and 95%, vs 95–97% for helical CT, while the specificity ranged between 85 and 98% for US and 94 and 97% for helical CT. Only one truly randomised study was found, showing that compared with US alone, a combination of US and helical CT increased the sensitivity from 86 to 99%, while the specificity decreased from 95 to 89%. We conclude that imaging should be performed in all children with suspected appendicitis and that US should be the initial procedure with CT as a complementary tool.