Professional role of the clinical pharmacologist.

The position of clinical pharmacologists in the field of medicine is reviewed. In more than one-third of the medical schools in Europe, teaching of clinical pharmacology is inadequate or non-existent. The situation in Italy is even worse; there are, however, 18 schools of specialization in pharmacology in that country, covering four areas, clinical and applied pharmacology, toxicology and chemotherapy with the course lasting four years. In addition there are 13 hospitals at regional level which provide service in clinical pharmacology, though in most of these the clinical pharmacologist has no direct clinical responsibilities. Three basic requirements, accepted by the College of Italian Pharmacologists, are listed aimed at obtaining official recognition of the professional role of a physician specialized in clinical pharmacology.


Establishing requirements for the use of terms for reporting adverse drug reactions.

The establishment of a project for the standardization of international reporting of adverse drug reactions (ADR), sponsored by the Council for International Organizations of Medical Sciences is described. The need to establish requirements for properly diagnosing a given suspected ADR and to describe it with the correct terms is demonstrated. However in view of the difficulties involved, it is suggested that the present deliberate approach needs to be reconsidered.


Efficacy and tolerability of medium-term treatment with simvastatin in primary hypercholesterolaemia.

3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, a new class of hypolipidaemic agents, have been recently shown to be effective in lowering both total and low-density lipoprotein (LDL)-cholesterol. In the present study 20 patients were treated with simvastatin for six months. Various dosing regimens of 10, 20, and 40 mg/day were employed. Administration of simvastatin 10 to 40 mg/day resulted in a 24.7 to 35.8% reduction in LDL-cholesterol levels and in a 19.5 to 26.9% decrease of total cholesterol levels. Moreover, simvastatin increased high-density lipoprotein-cholesterol and induced a decrease in plasma triglycerides. The side-effect profile was similar to other observations and included reversible transaminase elevations, myalgia and gastrointestinal complaints. From the present study simvastatin seems to be well tolerated in the short- to medium-term and to be effective; however, its long-term tolerability needs to be confirmed.


Effect of midecamycin acetate on gastrointestinal motility in humans.

The effect was studied on gastrointestinal motor activity of three different macrolides (erythromycin, roxithromycin and midecamycin acetate), administered by mouth in therapeutic doses. This placebo-controlled study was performed in 12 normal human subjects by means of intraluminal pressure measurements in the gastric antrum, duodenum and upper jejunum. In each subject, three manometries were done for 5 h in the interdigestive period and for 3 h postprandially. In the interdigestive period, midecamycin acetate did not affect the characteristics of the gastric migrating motor complex (MMC) and did not increase the number of antral contractions or the gastric motility index as compared to the placebo. Erythromycin and roxithromycin increased the number of antral contractions (24.5 +/- 11 versus 15.2 +/- 7 and 28.4 +/- 12 versus 14.9 +/- 5.9 respectively) and the motility index (4.05 +/- 0.5 versus 3.17 +/- 0.6 and 4.38 +/- 0.2. versus 3.64 +/- 0.7 respectively) as compared to the placebo. In the postprandial period, the number of antral contractions was not significantly increased by any of the three antibiotics. The postprandial antral motility index was not significantly increased by midecamycin acetate. In contrast, the postprandial antral motility indexes after erythromycin (4.4 +/- 0.5) and after roxithromycin (4.3 +/- 0.2) were significantly greater than after the placebo. In the upper small intestine, erythromycin elicited an increased number of phase-III-like activity events and roxithromycin shortened the MMC cycle length. Midecamycin acetate had no effect on interdigestive upper jejunal motility. The postprandial jejunal motor activity was not altered by any of the three antibiotics neither during the interdigestive nor the postprandial periods.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of calcium channel blockers on cardiovascular responses to smoking in normotensive and hypertensive smokers.

To evaluate the effects of short- and long-term administration of calcium (Ca) entry blockers on cardiovascular responses to cigarette smoking, 28 normotensive and 26 hypertensive smokers were studied. According to a double-blind, cross-over design, normotensive smokers were given a single dose (10 mg) of isradipine or a placebo and were checked before and after dosing. After a four-week period on a placebo, hypertensive smokers were treated with slow-release nicardipine 40 mg twice daily for six months and were checked at the end of the placebo period, after the first dose of nicardipine and at the end of six months of therapy. In both groups, blood pressure and heart rate were monitored by a Takeda TM 2420 non-invasive device every 3 min for 2 h. During the first hour patients were not allowed to smoke, during the second hour they were asked to smoke one cigarette every 15 min. In both normotensive and hypertensive smokers, calcium entry blockers produced a significant attenuation of the rise in blood pressure induced by cigarette smoking. Such effect was more evident after long-term therapy. Acceleration of the heart rate due to smoking was not significantly affected by the administration of Ca-antagonists, although a tendency towards a lesser heart rate increase was observed, particularly after long-term treatment. It was concluded that calcium entry blockers, reducing blood pressure variability, which bears a positive relationship with target organ damage, might be useful in the treatment of hypertensive smokers who were unwilling or unable to stop smoking.


Metabolic aspects of acute cerebral hypoxia during extracorporeal circulation and their modification induced by acetyl-carnitine treatment.

Following their previous research experiences in human tissue hypoxia, in the present study the authors. investigated the metabolic effects of acute brain hypoxia in a group of patients in course of extracorporeal circulation for aorto-pulmonary bypass. One hundred subjects were treated, half with a placebo and half with acetyl-carnitine to evaluate the effects of oxidative stress in some brain plasmatic metabolites and to verify the effect of acetyl-carnitine on the tissue energy capacity. The levels of lactate, pyruvate, succinate and fumarate showed a significant imbalance due to hypoxia, while the acetyl-carnitine treatment confined the metabolic gradients within physiological limits. This means that during the course of extracorporeal circulation brain hypoxia plays a pathological role assuming the typical picture of cellular oxidative damage and the acetyl-carnitine antagonizes these deleterious effects of hypoxia by a protective mechanism on the energy processes and then on the cellular enzymic activities. In this regard, the d-tyrosine levels, considered as a proteolytic index, confirm the action of acetyl-carnitine on the cell morpho-functional integrity.


Combined therapy with cyclobenzaprine and ibuprofen in primary fibromyalgia syndrome.

Non-steroidal antiinflammatory drugs are not very effective, even if commonly used, in primary fibromyalgia syndrome (PFS), whereas cyclobenzaprine (C) has proved to be quite useful. The aim of this open randomized study was to compare low-dose C alone or in combination with ibuprofen (I) in 32 female patients suffering from PFS. Fifteen patients were given C 10 mg, and 17 patients C 10 mg plus I 600 mg. All patients received the drugs orally at night and were evaluated at baseline and at days 5 and 10. Assessment of efficacy included the number of tender points (max. = 16), muscle tightness (score 1-5), sleep difficulty (score 1-10), pain intensity (visual analogue scale 0-10) and duration of morning stiffness (min). At the end of the study all symptoms were found to be improved to the same extent in both treatment groups. An exception was morning stiffness, which became significantly more reduced in the patients taking C plus I. No patient discontinued the trial owing to adverse side-effects. In conclusion, C and I given concomitantly at night proved to be safe and advantageous in relieving the discomfort of PFS in the short term.


Subchondral vascularisation and osteoarthritis.

A prospective experiment was carried out in 40 rabbits in which the blood supply to the patella was interrupted for defined periods from 2 weeks to 6 months. With prolonged ischaemia, there were clear changes in the articular cartilage which were comparable with those seen in human osteoarthritis.


Histological examination of surface repair tissue after successful osteotomy for osteoarthritis of the hip joint.

In this study biopsy specimens of the surface repair tissue from the weightbearing area of the hip joint were collected from 15 patients who had undergone successful valgus osteotomy, and these were examined histologically. In 6 of the 15 patients the specimens stained strongly with safranin-O in all three layers of the matrix. However, the repair tissue from another 6 patients showed strong staining only in the intermediate and deep layers, while that from 2 patients stained only in the deep layer. Specimens from 5 patients were stained for S-100 protein and positive cells were detected in all cases, demonstrating that the cells in the repair tissue were chondrocytes. However, there was no tide mark in the repair tissue and the arrangement of the cells differed from that in normal cartilage. Discontinuity of the subchondral bone was also noted. These findings make it likely that the repair tissue was of bone marrow origin.


The long term outcome of open total and partial meniscectomy related to the quantity and site of the meniscus removed.

Two hundred patients were randomly selected for either a partial or a total meniscectomy for a meniscal tear during open operation. They were followed for a median of 7.8 years after operation. After partial meniscectomy, posterior horn tears had the worst outcome, but this was only apparent when more than two-thirds of the meniscus had been removed. The amount of meniscal tissue excised was inversely related to the function of the knee, except with bucket-handle tears treated by partial meniscectomy. Patients with bucket-handle, anterior and posterior horn tears had similar functional results after total meniscectomy. Preservation of the peripheral rim of the meniscus following partial meniscectomy produces the best functional results.


Osteochondritis dissecans patellae. Is there a genetic predisposition?

The aetiology of Osteochondritis Dissecans Patellae remains controversial. We present a family suffering from the condition in all members of one generation. Clinical data support the hypothesis that the aetiology of the disorder in this family is recurrent subluxation of the patella, the inherited predisposing factor being an abnormality of the mechanics of the knee.


Synovial haemangioma of the knee joint.

Synovial haemangioma of joints are rare, but usually affect the knee joints of children and adolescents. Recurrent swelling, usually due to haemarthrosis, and intermittent pain may be present for a long time before the diagnosis is made. In this case report the common features of the condition are described, with emphasis on the occurrence of atraumatic haemarthrosis in children. The value of arthroscopy in diagnosis is discussed.


Intertrochanteric osteotomy for the treatment of chronic slipped capital femoral epiphysis.

Thirty six patients with a chronic slip of the capital femoral epiphysis and a femoral headneck angle of more than 30 degrees, as measured in a lateral radiograph, were treated by intertrochanteric osteotomy. The patients had an abnormal gait, an average age of 14.1 years and symptoms for an average of 14.5 months. The postoperative complications included two patients with avascular necrosis of the femoral head, four with coxa vara and two with loss of position requiring further operation. At an average of 7.5 years after operation 14 were good, 18 fair, and 4 poor on clinical assessment, with 13 good, 19 fair and 4 poor when judged by radiographs. Patients with complications had only fair or poor results. A postoperative head-neck angle of less than 10 degrees indicated a good result.


The management of unreduced traumatic dislocation of the hip in developing countries.

Traumatic dislocation of the hip often remains untreated in developing countries. This paper reports the outcome of twenty nine patients with neglected dislocations (3 anterior and 26 posterior). We have outlined the indications for five different methods of treatment. An attempt has been made to explain the causes of poor results.


The suppression of heterotopic ossification after total hip arthroplasty.

In a double-blind prospective randomised study we examined the effects of Diclofenac on heterotopic ossification after hip arthroplasty. Either the drug, or a placebo, was given by mouth to 158 patients in doses of 3 x 50 mg for 6 weeks. Diclofenac resulted in highly significant improvement (p less than 0.0001 versus controls) without severe side-effects. Heterotopic ossification decreased from 55% in the placebo to 15%. Significant degrees of heterotopic ossification did not occur with the drug. Movement of the hip was considerably increased after operation. We recommend Diclofenac prophylaxis against heterotopic ossification after hip operations in view of the serious clinical effects of this complication and its quoted incidence of 15% to 80%.


Experimental kyphoscoliosis induced in rats by selective brain stem damage.

The relationship between damage to particular brain stem nuclei related to postural control and equilibrium and the occurrence of spinal deformity has been investigated. The model used was the Wistar albino rat and the brain stem structures damaged were the gracillis nucleus, the superior colliculus and the lateral vestibular nucleus. Out of 60 brain stem damaged animals, 44 had accurate lesions from which 11 (25%) showed kyphoscoliotic deformities. Rats with spinal deformity showed an imbalance of the paraspinal muscles when assessed by EMG; this was expressed by an increase of muscular activity on the convex side. This study indicates that postural dysfunction caused by brain stem damage may induce kyphoscoliosis in experimental animals and could be one of the underlying mechanisms in the production of human idiopathic scoliosis.


Failed lumbar spinal surgery.

Failures and poor results of 160 patients after lumbar spinal surgery between 1980 and 1984 were analysed retrospectively. A self-rated questionnaire carried out 12 months after operation revealed 20 poor results; these occurred most commonly after multiple operations, decompression and fusion as compared to disc excision. The commonest cause was failure to recognise abnormal pain behaviour before operation. A more careful preoperative assessment should reduce the incidence of failure.


[Clinical results of the treatment of chronic hematogenous osteomyelitis of the femur by operation. A series of 165 cases].

Between 1968 and 1987 the authors treated 165 cases of chronic haematogenous osteomyelitis of the femur by operation. The procedure used combined decompression, sequestrectomy and saucerization in 161 cases, and sequestrectomy alone in 4. Chemotherapy was used in association for between 10 and 60 days. Eleven patients, thought to be healed, were lost to follow-up after 4 months. The remainder were reviewed for between 4 and 252 months, 124 for more than one year. After the first operation 105 lesions (64%) healed completely, while 60 (36%) relapsed. Of these, 48 were treated by further operation and 12 by drainage of the abscess. Nine patients relapsed twice and there were 3 or more relapses in a further 9. When last seen, 147 lesions were healed and 18 had a permanent sinus. Special features in relation to the femur are discussed, including destructive lesions of the knee and hip joints, the high incidence of stiffness of the knee even in the absence of radiographic evidence of destruction (48.5%), and the danger of injuries to the femoral vessels when using an approach through a medial sinus. The frequency of occurrence of lesions on both medial and posterior aspects of the femur has led the authors to recommend definitive surgical approaches, especially the true postero-lateral and antero-medial routes. Unsatisfactory results after operation may be due to the difficulty in performing sequestrectomy in deep and extensive lesions, problems in undertaking adequate saucerization and the lack of muscle to fill the residual cavity.


Evaluation of radiological investigations in spinal tuberculosis.

Thirty-four cases of spinal tuberculosis were investigated by plain radiography, computerised tomography and myelography followed by computerised tomography. All the patients were operated on and the findings compared with the results of the investigations.


Reconstruction of the pelvic ring after tumour resection.

Three different types of vascularised bone grafts have been used to reconstruct the pelvic ring after resection of tumours. Rapid bone healing is guaranteed and the period of rehabilitation is lessened compared with conventional methods.


Intraoperative radioactive localization of small bone tumours.

A sodium-iodide scintillation probe and a portable scaler rate-metre were used to localize small bone tumours at operation, after the preoperative injection of technetium-99m methylene diphosphonate. This technique allows complete removal of the tumour nidus of an osteoid osteoma or the active part of other lesions without undue sacrifice of the surrounding bone. The method has been used for successful relief of pain in thirteen patients.


Allogenic decalbone in the repair of benign cystic lesions of bone.

Forty-six cases of benign cystic lesions of bone were treated by curettage and compact filling using partially decalcified allogenic bone graft (Decalbone); of these, 35 were available for study. Decalbone was prepared by partial decalcification with 0.6 N hydrochloric acid (HCl) of human bones generally obtained from freshly amputated limbs. The commonest lesions of bone were giant cell tumours (14) and aneurysmal bone cysts (15), and the commonest bones involved were the femur (23) and the tibia (12). There was one failure and four recurrences. Five cases were infected but this did not interfere with healing of the primary lesion. Radiological incorporation of the graft was seen at about 3 months in unicameral bone cysts, at 4-6 months in aneurysmal bone cysts and at 6-9 months in giant cell tumours. There was no recurrence in any case of giant cell tumour, but three aneurysmal bone cysts recurred. There was no clinical immune reaction.


Osteofibrous dysplasia of the tibia treated by bracing.

Osteofibrous dysplasia of the tibia in children will commonly recur after surgical resection. The use of a leg brace until puberty offers a useful method of conservative management. Seven female patients, age three months to nine years at the time of brace treatment, have been followed for 3.5 to 18 years, an average of nine years, with evidence of satisfactory healing of the lesions in all. Three of the cases had recurred after surgery, two with fibular grafts and one required leg lengthening. The use of a brace to control bowing of the tibia while awaiting spontaneous regression is advised until epiphyseal closure.


Leiomyosarcoma of the great saphenous vein.

Primary malignant tumours of the great saphenous vein are extremely rare with only 15 cases reported in the English literature since 1919. We report a patient who survived 5.5 years after treatment with surgery and chemotherapy before dying from lung metastases.


Arthrolysis in the relief of post-traumatic stiffness of the elbow.

A retrospective clinical study of 34 patients treated by arthrolysis for elbow stiffness is presented. These patients had sustained previous injury which was a simple fracture in 44%. The average age at operation was 31 years with a range from 6 to 57 years. Follow up for a minimum of one year, and an average of four years, showed an increase of motion from a preoperative average of 45 degrees to a postoperative average of 92 degrees with few complications and only three unsatisfactory results. This analysis has allowed conclusions concerning the technique and indications for operation.


Simultaneous flexion of the thumb and index finger after a fracture of the first metacarpal bone. A case report.

We report a case in which simultaneous flexion of the thumb and index finger occurred after a fracture of the 1st metacarpal bone. This was demonstrated to be due to a fibrous adhesion around an anomalous slip between the tendons of flexor pollicis longus and flexor digitorum profundus indicis.


[Walking in children after poliomyelitis].

Between 1983 and 1990 the authors examined 630 children who had suffered from poliomyelitis, mostly between the age of one and three, at an average age of 9 years 3 months. Half were unable to walk because of residual paralysis. Operation was carried out on 252 children. Most underwent soft tissue release at the hips, knees or tendo Achilles. Triple arthrodesis was carried out on 24. Calipers, produced locally, were used, and trunk bracing was not employed. The average duration of treatment was 3 months, with gradual achievement of correction. Vascular complications occurred in one leg, there were 5 paralyses of the sciatic nerve and 4 fractures of the upper tibia. One hundred children who were not able to walk before operation were reviewed. Sixty five were walking and able to go to school. The best results were achieved before the age of 12.


Bilateral ankylosis of the hips following heterotopic ossification of the ilio-psoas in a child.

A case of bilateral ankylosis of the hip in a 3-year-old child due to heterotopic ossification of the iliopsoas following a long period of coma is presented. A review of the literature has failed to reveal a similar case with such extensive heterotopic ossification. The patient was treated successfully by surgical excision of the heterotopic bone which was extra-articular and extra-capsular and involved only the iliopsoas muscles.


Pulmonary function in adolescents with idiopathic scoliosis.

Two groups of patients, with an average age of 15 years, have been studied to establish the improvement in pulmonary function after surgical correction in relation to the amount of correction achieved. The first group of 33 patients were treated surgically; an average Cobb angle of 72 degrees was reduced to 32.6 degrees. The second group of 30 patients with an average Cobb angle of 33 degrees were not operated on. Results of tests undertaken have shown that pulmonary function was improved by surgical correction, but the improvement does not match the degree of correction achieved. Statistical analysis of the two groups shows a difference in cardiopulmonary function to the advantage of those who were not operated on. This suggests that there is partial irreversibility of ventilation and perfusion in those who are treated surgically.


Correction of the sagittal plane in idiopathic scoliosis using the Zielke procedure (VDS).

This paper reports a retrospective review of 91 patients with idiopathic thoracic scoliosis corrected by Zielke VDS instrumentation. The results were analysed to demonstrate the correction in all three planes. Satisfactory correction was obtained, and was particularly good in the sagittal and horizontal planes. There were no disadvantages from the thoracotomy.


Operative treatment of unstable thoracolumbar fractures by the posterior approach with the use of Williams plates or Harrington rods.

Two similar groups of patients with unstable fractures and fracture dislocations of the thoracic or lumbar spine have been treated with the posterior application of Williams plates or Harrington distraction rods over a period of 10 years. The instrumentation was routinely removed from 13 to 19 months after insertion. The initial preoperative kyphotic angle was corrected by a mean of 4.5 and 6.9 degrees respectively. However, follow up after a minimum of four years has shown a loss of correction by a mean of 14.4 and 12.7 degrees to an angle of 21.9 degrees even in the Harrington rod cases. These results do not show the ability to maintain correction of deformity by either method with a dorsal approach. Reduction and stabilization of fractures of the spine should be directed to the anterior part of the vertebrae to overcome late deformity of the spine.


Translaminar screw fixation of the lumbar spine.

Lower lumbar spinal fusion was carried out in 72 patients by a technique which involved screw fixation of the facet joints. Stability was achieved and mobilisation was allowed immediately. Radiographs after operation showed bony fusion in 94.5%. Pain was diminished and 76% of the patients would undergo the same treatment again. There were no neurological complications. The technique is useful and safe for the fusion of short segments of the lumbar spine.


Halo-cast treatment of fractures and dislocations of the cervical spine.

Twenty-eight consecutive patients with dislocated, and often unstable, injuries of the cervical spine were treated by halo-cast stabilisation. One died and 27 were followed up. Twenty had no symptoms and 17 had full movement of the neck. There were only a very few minor complications. Seven patients had initial neurological impairment. They included the single death, but the remaining 6 regained useful muscle function. The halo-cast allows complete reduction in many types of fractures and dislocations, and good immobilisation is maintained.


Transplantation of an island fascial flap with a vascular pedicle for arthoplasty of the hip.

Intra-articular transplantation of a vascularised island fascial flap has been used for arthroplasty of the hip in 18 patients with an average age of 22 years. They have been followed up for an average of 3 years; 14 were graded excellent and 4 good using the criteria of Charnley and Wu Zhikang. The procedure has a place in the treatment of hip disease in the younger patient.


Listeria monocytogenes infection in prosthetic joints.

Listeria monocytogenes is increasingly recognised as a cause of prosthetic joint infection. These infections tend to be more insidious and indolent in contrast to similar infections with Staphylococcus aureus. They can occur in patients who are immunocompromised due to malignancy or other illness or in nonimmunocompromised elderly patients. Listeria infections should not be treated with cephalosporins and prolonged antimicrobial treatment is generally advised for prosthetic joint infections. We noted a high affinity of L. monocytogenes for foreign bodies. Three of twenty-seven cases 11%) of listeriosis under the period of investigation were associated with foreign bodies (hip prosthesis, knee prosthesis and arterial graft).


Rotator cuff tears in anterior dislocation of the shoulder.

We diagnosed 16 rotator cuff tears in 109 shoulders in 104 patients with a traumatic anterior dislocation. Twelve of these were followed up for an average of 32 months. A surgical repair of the cuff was carried out in eleven and the results were satisfactory in eight (73%) regardless of a Bankart lesion. The repair of a torn cuff seems to be sufficient to stabilise the shoulder in elderly patients, even when a Bankart lesion is present and is not repaired.


Macrodactyly of the hands and feet.

Sixteen patients with macrodactyly have been reviewed; the hand was involved in 9 (one was bilateral) and the feet in 7. The condition was familial in 3. No qualitative difference was found between the static and progressive types. In the hand, the digital nerve was enlarged and lengthened whereas in the foot it looked normal. Microscopy showed infiltration of the nerve by fat and fibrous tissue in the hand and only slight perineural fibrosis in the foot.


[Absorbable rods and screws: a new method of fixation for fractures of the olecranon].

Between April 1986 and July 1990 fractures of the olecranon in 41 adult patients were treated by fixation with absorbable rods (20 patients) and screws (21 patients) of self-reinforced polyglycolide (SR-PGA), 3.2 mm in diameter and 20-70 mm in length. Patients were followed up for a mean time of 2 years 7 months (range 1 year to 4 years 6 months). After reduction of the fracture, channels were drilled from the proximal fragment through the cortex of the distal fragment and the fractures were fixed with absorbable rods or screws. By one year from follow-up maintenance of an anatomical reduction of the fracture was seen in 34 patients. Failure of fixation requiring a second operation occurred in 2 cases. In all cases functional recovery was at least satisfactory. Sinus formation as a sign of transient tissue reaction was observed in 3 cases, but did not influence the healing of the fractures or the functional recovery. The results in patients treated with rods or screws was similar. Absorbable screws combined with small rods and absorbable sutures allow treatment of +ore severe fractures of the olecranon than do rods alone.


Injuries in paragliding.

In a retrospective study, 376 paragliding accidents have been analysed. Leg injuries were most common, but a large number of spinal injuries also occurred. The causes were either misjudgement by the pilot or the influence of weather and terrain. Improvements in the instructor's knowledge and the pilot's training could have prevented most of the accidents. Analysis of the mechanisms of the crashes and the pattern of trauma help to produce an efficient approach to diagnosis and treatment.


Prediction of late complications of femoral neck fractures by scintigraphy.

A prospective study was undertaken to determine the value of scintigraphy in predicting nonunion or necrosis of the femoral head following femoral neck fractures. Scintigraphy was carried out in 49 patients before operation and in 37 one week after operation; in 18 it was done before and after operation. The uptake of isotope was estimated visually as either normal or reduced compared to the opposite side. All the patients were followed up for at least 2 years. The results indicate that pre-operative bone scintigraphy is useful in elderly patients to predict nonunion and necrosis, and therefore is recommended to select those patients who will benefit from primary arthroplasty instead of fixation of the fracture.


Fixation of fractures by twisted plates. A new concept of osteosynthesis.

A new twisted plate has been designed for biplane fixation of diaphyseal fractures of long bones. Comparative mechanical testing of the new twisted plate and conventional flat plates showed that the twisted plate was 40% stronger when a bending moment was applied and 132% stronger to a twisting moment. The overall improvement in strength was 90.5%. Clinical trials have been carried out and this paper describes the use of the new plate.


The deltoid muscle: an electromyographical analysis of its activity in arm abduction in various body postures.

We have carried out an electromyographical examination of the activity of five different regions of the deltoid muscle during abduction/adduction in various body postures with different biomechanical actions of arm gravity. The results show that the deltoid action is highly differentiated in its different regions and is not restricted only to the generation of an abducting moment in the shoulder joint. There is obviously a biomechanical contribution, mainly by its spinal and clavicular regions, to the stabilization of the glenohumeral joint and to the control of the selected plane of abduction.


The diagnostic significance of anti-type II collagen antibody assay in rheumatoid arthritis.

An improved enzyme-linked immunosorbent assay (ELISA) for the detection of anticollagen antibodies in human serum has been developed. With the use of this method, antibodies against native human Type II collagen were detected in 22.7% of sera from 480 patients with rheumatoid arthritis (RA). The antibodies were found to be collagen type specific, showing no reaction with human Type I and Type III collagens. The antibodies appeared in high incidence during the early phase of the disease, and RA patients with involvement of a single joint, mono-articular arthritis, were often positive for anti-Type II collagen antibodies. In most of these patients, anti-Type II collagen antibodies preceded the appearance of rheumatoid factors. The antibodies were all negative in sera from patients with gout, osteoarthritis (OA) and non-arthritic diseases. Thus, anti-Type II collagen antibody assay may have diagnostic significance for RA patients, especially those in whom laboratory and clinical findings provide only minimal help in diagnosis.


A new method for measurement of femoral anteversion. A comparative study with other radiographic methods.

A new biplanar method of measuring femoral anteversion, which may be considered a modified Magilligan's method, is described. In addition to the true antero-posterior radiograph, a true translateral radiograph is taken, instead of a transcervical lateral radiograph as in the Magilligan method. The acute angles (alpha and gamma) between the long axes of the shaft and neck of the femur on both the antero-lateral and lateral radiographs are measured. The angle (gamma') of anteversion is obtained by the trigonometric formula; tan gamma' = tan/tan alpha. The femoral anteversion in 20 adult dried femora and 40 femora in 20 children was measured by the Magilligan method, the authors' method and the conventional CT method. Compared with direct measurement in the dried femora, it was found that both the Magilligan and the authors' methods tended to overestimate and the CT method tended to underestimate (p < 0.05) the anteversion. In children's femora, a similar trend was observed among the three methods and the measurements by each method differed significantly from the other (p < 0.05). We feed that our method of biplanar measurement of femoral anteversion is more accurate, although probably marginally, than the conventional Magilligan method. It also has some relative advantages in clinical application in that positioning is easier, rotation is better controlled, and less time is consumed.


Patellar metastasis from a malignant melanoma.

We report a patient with a patellar metastasis from a malignant melanoma. Only 16 other cases have been previously described in detail in the literature. A lytic lesion in the patella is most often caused by a benign tumour, but a primary or secondary malignant tumour has to be excluded.


Comparison of magnetic resonance imaging and computed tomography in the local assessment of osteosarcoma.

We have compared the value of magnetic resonance imaging (MRI) with computed tomography (CT) in assessing the extent and the localization of osteosarcomas. Twenty patients were examined by both systems, and the results related to the clinical and histological findings. While both investigations are valuable, MRI may be more useful for defining the local extent of an osteosarcoma.


One session surgery on the lower limb in children with cerebral palsy. A five year follow-up.

Twenty three children with cerebral palsy, who were operated on upon before 1984, have been re-assessed. All the lower limb deformities were corrected in one surgical session. Review after five years included foot-switch measurements and assessment of video recordings. There were no recurrences, and gait ability showed normal growth-related development.


Morton's neuroma.

During the past 10 years we have operated on 75 feet for Morton's neuroma with a minimal follow up of nine months. Associated anomalies were found in 80%. Pathological studies demonstrated that the lesion was irritative in nature. The clinical results were satisfactory in 94%.


Humoral immune responses in osteomyelitis.

Fifty-three patients with osteomyelitis were grouped according to the duration of the disease at presentation, the bone and the part involved, the aetiology, activity, severity and stage of disease. Sera from these patients was estimated for IgA, IgM and IgG levels and in 23 matched controls. The results were compared with the normal range, and correlative studies were carried out between serum immunoglobulin levels and the different clinical parameters. Serum levels of the 3 immunoglobulins were found, in general, to fail to rise above the normal in the presence of infection. Furthermore, the levels did not show significant variation with the clinical parameters. These findings suggest that there is an intrinsically subdued immune response to osteomyelitis in most of the patients, partly confirming the suspicion of an immunodeficient state.


Septic arthritis in HIV positive haemophiliacs. Four cases and a literature review.

Septic arthritis is rare in haemophiliacs. Four new cases who were also HIV positive are reported. In three, the knee was involved, and in the fourth the elbow. The organism was streptococcus pneumoniae and staphylococcus aureus in one patient each, and salmonella in two. Although all the patients were human immunodeficiency virus (HIV) positive at the time of diagnosis, only two patients developed autoimmune deficiency syndrome (AIDS) after their septic arthritis. These two died later due to AIDS complications. Treatment was conservative in all cases with antibiotic therapy and prompt rehabilitation. The results were fair in two and good in two. Therefore nonoperative management is advocated before surgical drainage is considered. It seems likely that a positive HIV status is related to the appearance of septic arthritis in haemophiliacs.


Orthopaedic complications in sickle cell disease. A comparative study from two regions in Saudi Arabia.

The haematological and orthopaedic complications of patients with sickle cell disease from two different regions of Arabia are presented. Nineteen patients from the South-western region were matched for age and sex with an equal number from the Eastern region. The mean sickle cell haemoglobin was 75.95% in those from the South-west and 77.5% in those from the East. The haemoglobin concentration was marginally lower in the Eastern region patients at 9.19 g/% compared with 9.51 g/%. Bone and joint infections occurred in 17% of Eastern region patients and in 15% in those from the South-western region. There was no significant difference between the haematological parameters and the orthopaedic complications in the two regions. We conclude that these complications are severe in the Eastern region and the disease is not benign as previously thought. Aggressive treatment of orthopaedic complications is indicated in sickle cell disease in the Eastern region.


"Existential" therapists and their group therapy practices.

Existential philosophical thought insists that human behavior is indeterminate and subject only to the individual's will. It is, therefore, skeptical of all dynamic psychologies based as they are on Freudian determinism. Group therapists imbued with this philosophy stop short of its extreme. Their existential position, however, does greatly modify treatment, and yet they preserve both the form and goals of therapy. They place properly selected and prepared patients in a group where their individuality and authenticity are highly valued. Finding most technical procedures irrelevant, the therapists themselves, that is, their evolving personalities are central. The key to patient change is the spontaneous meeting of members and the therapist which the therapist orchestrates. Instead of the usual interpretation, members are encouraged to confront the paradoxes in their lives, their humanness, and especially their finitude. Patients are to be brought up to the threshold of their self-knowledge so they can choose. Choice, therefore, along with action coupled with responsibility are frequent themes. Unless the individual is incompetent, decisions made for him or her by the therapist or by group consensus are thought to be nontherapeutic.


Group-analytic psychotherapy.

Group analysis is a flexible and effective method of providing group psychotherapy, thereby promoting maturation and symptom relief. Its metapsychology remains incompletely worked out but provides potentially exciting interfaces with biological and physical sciences, through Foulkes's (1973) seminal concept of the group matrix.


Psychodrama: group psychotherapy through role playing.

The theory and the therapeutic procedure of classical psychodrama are described along with brief illustrations. Classical psychodrama and sociodrama stemmed from role theory, enactments, "tele," the reciprocity of choices, and the theory of spontaneity-robopathy and creativity. The discussion focuses on key concepts such as the therapeutic team, the structure of the session, transference and reality, countertransference, the here-and-now and the encounter, the group-as-a-whole, resistance and difficult clients, and affect and cognition. Also described are the neoclassical approaches of psychodrama, action methods, and clinical role playing, and the significance of the concept of behavioral simulation in group psychotherapy.


Feminism and group psychotherapy: an ethical responsibility.

In response to Martin Lakin's (1991) IJGP article, "Some Ethical Issues in Feminist-Oriented Therapy Groups for Women," this article examines recent developments in feminist theory and proposes that a feminist perspective is both ethical and can make significant contributions to the practice of group psychotherapy. The overview of feminist theory focuses on (1) the importance of the social context, (2) contributions and challenges to psychoanalytic and developmental theory, (3) attention to power relations, (4) the connection between the personal and political, and (5) recognition and integration of diversity and difference. Clinical examples illustrate ways in which male and female group therapists can take a feminist perspective and become "ethical advocates."


Modulation of IL-2, IFN-gamma, TNF-alpha and IL-4 production in mice of different ages by thymopentin.

The effect of an immunomodulator drug thymopentin (TP5) on the production of various cytokines (IFN-gamma, IL-2, IL-4, TNF-alpha) in mice of different ages has been studied. TP5 enhanced IL-2, TNF-alpha and IFN-gamma production but reduced the IL-4 secretion by splenocytes from aged mice (greater than 120 week old) in vitro. However, it had no effect on the IL-2, IFN-gamma, TNF-alpha or IL-4 production by splenocytes from young and adult mice. TP5 injected subcutaneously was able to induce high levels of IL-2 production by splenocytes from all groups of mice. The TP5 effect on TNF-alpha and IFN-gamma was similar, even though it was significant only in old mice. Furthermore, TP5 was able to significantly reduce IL-4 production in old mice, which normally produced high levels of this cytokine after mitogen stimulation. Since it has been observed in the mouse that the Th1 cells secrete IFN-gamma and IL-2, whereas the Th2 cells preferentially produce IL-3, IL-4 and IL-5, these results indicate that the immunopotentiatory activity of TP5 is due to the preferential up-regulation of Th1 cells.


Low dose streptozotocin-induced diabetes in mice: reduced IL-2 production and modulation of streptozotocin-induced hyperglycemia by IL-2.

The possible role of interleukin 2(IL-2) in the pathogenesis of multiple low dose streptozotocin (Sz)-induced diabetes in mice was analysed. Spleen cells from diabetic male C57Bl/6 mice showed diminished mitogen-induced IL-2 production as determined by bioassay using the IL-2-dependent T-cell line CTLL-2. In parallel the proliferative response was reduced. Systemic daily administration of human recombinant IL-2 for 3 weeks had dose-dependent effects on the development of hyperglycemia in Sz-treated (5 x 40 mg) mice: while IL-2 at doses of 1 x 2, 1 x 10, 2 x 10 micrograms/kg body weight caused partial suppression of hyperglycemia, higher doses (2 x 20, 2 x 40 micrograms/kg) had an enhancing effect. Treatment with the lowest dose (1 x 1 micrograms/kg) or with a control preparation from bacteria (2 x 10 micrograms/kg) did not significantly alter the course of diabetes. Effects of IL-2 were similar when treatment was started concomitantly with or only after streptozotocin injections. This observation argues against the direct interaction between IL-2 and streptozotocin but suggests modulation of immune reactivity by IL-2. Our findings of decreased mitogen-stimulated IL-2 production by splenic lymphocytes, and the disease-modulating effect of IL-2 in the low-dose streptozotocin diabetes extend our previous observations in spontaneously diabetic BB rats and further support the notion of an involvement of IL-2 in the control of autoimmune diseases.


Differential sensitivity of in vivo TNF and IL-6 production to modulation by anti-inflammatory drugs in mice.

The effect of dexamethasone and two non-steroidal anti-inflammatory agents ibuprofen and indomethacin on the production of serum interleukin 6(IL-6) and tumor necrosis factor (TNF) levels in mice treated with endotoxin (2.5 micrograms/mouse, i.p.) was investigated. Pretreatment of mice with dexamethasone (0.3-30.0 mg/kg, i.p., 30 min before endotoxin) completely blocked TNF production but did not affect that of IL-6. Conversely, pretreatment with indomethacin (5 mg/kg, i.p.) or ibuprofen (30 mg/kg, i.p.) potentiated the production of both IL-6 (+ 80% with INDO; + 100% with IBU) and TNF (+ 500% with INDO; + 50% with IBU). In the case of IL-6, the two anti-inflammatory drugs were able per se to induce significant levels of this cytokine even in the absence of LPS. These data indicate that IL-6 and TNF production are differently susceptible to glucocorticoids, and that prostaglandins can physiologically provide a negative feedback regulation of IL-6 and TNF synthesis.


The in vitro effects of mercury on peritoneal leukocytes (PMN and macrophages) from inbred brown Norway and Lewis rats.

The present paper demonstrates that HgCl2 can affect rat peritoneal polymorphonuclear leukocyte (PMN) and macrophage (M phi) functions in vitro. In addition, we have noticed that these effects of mercury vary according to the rat strain: for example, HgCl2 stimulates H2O2 release from Lewis (LEW) but not Brown Norway (BN) PMN. Similarly, LEW M phi produce high levels of H2O2 when exposed to HgCl2 in vitro, whereas BN M phi do not. Finally, mercury inhibits erythrophagocytosis of both LEW and BN "resident" peritoneal M phi. Preliminary experiments using M phi from other rat strains have also shown that MAXX M phi are stimulated by HgCl2 to release H2O2 in vitro, whereas Yoshida M phi are inhibited. Differences in lymphocyte responses (e.g. delayed-type hypersensitivity reactions and mitogen stimulation) between rats of various strains are well known. To these examples one may now add variations in PMN and M phi responses to mercury and possibly other metals. Our results suggest that caution should be exercised in interpreting the outcome of immunotoxicity studies in experimental animals. In particular, outbred rats may not provide appropriate models, that might be better obtained by comparative investigations of rats from various inbred strains.


In vitro activation of murine peritoneal exudate cells (PEC) and peritoneal macrophages by ST 789.

ST 789 is a new synthetic compound characterized by an amino acidic group joined to the N9 position of the hypoxanthine ring, which has been shown recently to have immunomodulating properties and minimal toxicity. The drug has been reported to protect immunosuppressed mice from microbial infections and tumour growth, and to restore the mitogen-induced proliferation of splenocytes from immunosuppressed young mice. In this study, we show that in vitro addition of ST 789 is able to markedly augment the sheep red blood cells (SRBC) phagocytosis by PEC, and to potentiate the cytotoxic activity of peritoneal exudate (PE) macrophages (M phi) vs the L-M tumour cell line. We also found that ST 789 enhanced the rIFN-gamma-induced NO2- release from cultured PE M phi. Similarly, in vitro addition of ST 789 to the latter cultures significantly increased the production of interleukin 1 (IL-1) and tumour necrosis factor (TNF) induced by lipopolysaccharide (LPS). These studies demonstrate that ST 789 is a potent phagocyte activator for the induction of cytokine release, phagocytosis and cytotoxic activity against tumour cells in vitro.


Antiviral activity of individual versus combined treatments with interferon alpha, beta and gamma on early infection with HTLV-I in vitro.

We have shown previously that infection of mononuclear cells derived from neonatal cord (CBMC) or adult peripheral (PBMC) blood with HTLV-1 can be controlled in vitro by treatment with interferon (IFN) alpha, beta or gamma. The activity of IFNs was mainly related to the induction of an active antiviral competence in host's immune effector cells. The antiviral activity of IFN-boosted CBMC could be ascribed both to a positive regulation of cell-mediated immunity and to inhibition of viral infection. Data described herein provide further information on the mechanisms of the antiviral activity of IFNs and compare the activity of each type of IFN with the association of alpha + beta, alpha + gamma and beta + gamma IFNs, at a concentration of 100 or 1000 IU/ml. When added at the onset of the co-culture of CBMC with lethally irradiated, virus-donor MT-2 cells, IFNs could protect host CBMC by inhibiting HTLV-1 infection in terms of reduced proviral integration and a lower percentage of virus-positive cells, until 4 weeks of culture. Infection of CBMC was inhibited at a comparable extent by either individual or combined IFN treatments. However, a clearcut inhibition of HTLV-I transcription was found only when alpha 100 + beta 1000 IU/ml and especially alpha 1000 + gamma 100 IU/ml combined treatments were tested. When the chronically infected, virus-producing MT-2 cells were treated with IFNs, a remarkable inhibition of HTLV-I transcription was found only after multiple treatments. However, MT-2 cells became resistant to the antiviral activity of IFN gamma, but not to that of IFN alpha or beta. These data provide further information on the control of HTLV-I replication mediated by IFNs at different steps of the viral life cycle, being therefore relevant to the clinical use of combined IFNs in the treatment of acute infection. Moreover, IFNs could be used to prevent the establishment of a persistent infection, which is a prerequisite for developing adult T-cell leukemia (ATL) and/or virus-associated myelopathy.


Spermine dialdehyde, a novel ex vivo purging agent for both allogenic and autologous bone marrow transplantations.

The use of ex vivo purging agents has shown to be beneficial for both allogeneic and autologous bone marrow transplantations. We have shown previously that spermine dialdehyde (SDA), an oxidized product of spermine, when used in phosphate-buffered saline (PBS) preferentially inhibits T-cell proliferation while sparing myeloid cells. Lethally irradiated mice were rescued by reconstitution with SDA-treated allogeneic marrow and showed no sign of graft-vs-host disease (GVHD). In this paper, we show by HPLC analysis that SDA degraded rapidly in PBS but remained intact in saline. Administered in saline, SDA was more inhibitory on leukemic cell lines than normal myeloid or T-cells. Lethally irradiated mice receiving a syngeneic bone marrow leukemic cell mixture treated ex vivo with SDA in saline did not manifest leukemia. Thus, the preferential inhibitory effect of SDA and its degraded products in different buffers suggest that SDA could act as a novel purging agent for both allogeneic and autologous bone marrow transplantations.


Effects of K-76COOH (MX-1) on immune response: induction of suppressor T-cells by MX-1.

K-76COOH (MX-1), isolated from the cultured supernatant of a species of fungi imperfecti, Stachybotrys complement nov. sp. K-76, is an inhibitor of the complement component, C5. The effects of MX-1 on various immune responses were investigated. MX-1 enhanced the response of spleen cells to PHA and LPS: MX-1 at 0.01-250 micrograms/ml for PHA and at 10-250 micrograms/ml for LPS. In contrast, it inhibited the response to Con A: MX-1 at 0.01-500 micrograms/ml for spleen cells and at 100-500 micrograms/ml for thymocytes. MX-1 and IL-1 synergistically acted to enhance the Con A response of spleen and thymus cells from which accessory cells and Ia-positive cells had been removed by passing through Sephadex G-10 columns and treating with anti-Ia monoclonal antibody plus complement. T-cells pretreated with MX-1, IL-1 and Con A for 3 days suppressed not only the response of B-cells to LPS but also the production of anti-SRBC antibodies. In addition, MX-1 was found to increase CD8+ T-cells. These results suggest that MX-1 acts on T-cells to induce suppressor T-cells.


Effects of subcutaneous versus intravenous administration of cyclosporin A on rat thymic histology and pharmacokinetics.

Cyclosporin A (CsA) was administered to rats by repeated subcutaneous (s.c.) or intravenous (i.v.) injections for 14 days. Changes in thymic histology were independent of the route of administration. Blood concentration--time profiles of CsA were similar at dose levels of i.v. 7.5 and s.c. 15 mg/kg/day, respectively. So, provided that the dose is reduced, i.v. injection can serve as an adequate alternative to s.c. injection, thereby preventing unwanted painful side-effects associated with the latter route of administration.


Differential effect of dipyridamole upon thymidine incorporation and expression of surface activation antigens by in vitro stimulated lymphocytes.

Dipyridamole inhibits, by 50% at 10(-6) M and 100% at 10(-5) M, the incorporation of tritiated thymidine by lymphocytes stimulated with 1 microgram/ml phytohaemagglutinin A for 72 h. At the latter concentration, a clinically relevant one, dipyridamole fails to inhibit lymphocyte proliferation and expression of the cell surface stimulation markers, CD25 and HLA-DR, or differential regulation of the CD45 -RA and -RO isoforms, as determined by single laser two-colour flow-cytometry. No significant immunodepressant effect of dipyridamole can be demonstrated in vitro.


Cocaine-induced suppression of interferon-gamma secretion in leukocytes from young and old C57BL/6 mice.

The effects of cocaine (0.1-100 micrograms/ml) were studied on interferon-gamma (IFN) secretion in Con A-stimulated splenocytes from young (4 months) and old (18 months) C57BL/6 mice. IFN secretion was significantly suppressed by cocaine in a concentration-dependent manner. Suppression was observed in splenocytes from both young and old mice. Splenocytes from young mice released higher amounts of IFN (23.40 +/- 1.47 ng/ml) than those from old mice (6.10 +/- 0.35 ng/ml) after 24 h in culture. The concentration of Con A used to stimulate IFN secretion significantly affected the suppressive effect of cocaine. Pretreatment of splenocytes with cocaine followed by culture in the absence of cocaine did not affect IFN secretion in old mice, while splenocytes from young mice showed a sustained depression in IFN release. This suggests that cocaine suppresses IFN release in vitro and the susceptibility to this inhibitory effect may be age related.


Effect of apocynin on the induction of ulcerative lesions in rat skin injected with tubercle bacteria.

Apocynin is an effective and selective inhibitor of the neutrophil oxidative burst in vitro. Other neutrophil functions, tested in vitro, such as chemotaxis, exocytosis, phagocytosis and intracellular killing of bacteria are unaffected by apocynin. These properties make apocynin a potential anti-inflammatory agent in vivo. This was tested in WAG/Rij rats, injected intracutaneously with tubercle bacteria in oil (complete Freund's adjuvant). We show here that continuous administration of apocynin via the drinking water prevents the formation of ulcerative lesions in the inflamed skin. No effects on humoral and cellular immunity were observed after treatment with apocynin (between 1 and 100 micrograms/ml drinking water) measuring serum antibodies and delayed-type hypersensitivity, respectively.


Immunosuppression of triptolide and its effect on skin allograft survival.

In this study the immunosuppressive properties of triptolide were evaluated. Triptolide was found to inhibit skin allograft rejection in a dose-dependent manner. This inhibitory effect was time dependent. Triptolide at 0.1 mg/kg/day significantly prolonged the graft survival when triptolide was given for 9 days after transplantation, but not before transplantation. In vitro studies showed that triptolide markedly suppressed cytotoxic T-lymphocyte (CTL) induction and mixed lymphocyte reaction (MLR) at concentrations ranging from 0.08 to 10 ng/ml. The inhibition on MLR was also significant when triptolide was added to the cultures at 36 h after initial incubation. Furthermore, exogenous IL-2 did not reverse this inhibitory effect of triptolide. Our results suggest that triptolide inhibits lymphocyte activation at a relatively late stage, and its effect on immune response is not exerted through altering IL-2 production.


Stimulation of microbicidal host defence mechanisms against aerosol influenza virus infection by lentinan.

The ability of polysaccharide immunomodulator lentinan to stimulate non-specific resistance against respiratory viral infections was investigated. Significant protection was conferred by lentinan administered intranasally before lethal influenza virus infection and could be corroborated by a reduction of the lung virus titres. Since the lung is the target organ of influenza virus infection, lentinan was also administered by the intravenous route. Lentinan conferred complete protection against a LD75 challenge dose of virulent influenza virus and significantly prolonged the survival time after a LD100 challenge. The effect on respiratory burst of broncho-alveolar macrophages was investigated by luminol-dependent chemiluminescence (CL) in response to stimulation by zymosan. Enhanced CL activity was present at an early stage in groups receiving lentinan. Significant nitric oxide activity could also be stimulated by culturing broncho-alveolar macrophages in the presence of lentinan. TNF activity could not be detected in lung lavage but measurable IL-6 was produced already after 6 h in animals administered lentinan alone and in lentinan-pretreated influenza virus-infected mice. Influenza virus alone did not induce measurable IL-6 at 6 h but high activity was present at later time periods.


Immunopharmacological properties of azepino [2, 1-b] quinazolin-12(6H)-one-7, 8, 9, 10-tetrahydro (RLX).

The effect of azepino [2,1-b]quinazolin-12(6H)-one-7, 8, 9, 10-tetrahydro (RLX) was studied on cell-mediated and humoral components of the immune system. In single (50, 100 and 200 mg/kg) and multiple (6.25, 12.50, 25 and 50 m/kg) oral dose schedules RLX significantly reduced the early (4 h) and delayed (24 and 48 h) hypersensitivity reactions to SRBC in mice. Daily oral administration of RLX (25, 50 and 100 mg/kg) produced a dose-related reduction in developing adjuvant arthritis in rats. In carrageenan-induced pleurisy in rats RLX (25, 50 and 100 mg/kg p.o.) caused a marked reduction in the volume of pleural exudate (25.26-59.64%) and infiltration of leucocytes (28.75-50.29%) into the pleural cavity. RLX produced marked inhibition of humoral antibody synthesis both in mice and rats and that of complement fixing antibody in mice. It was not cytotoxic and showed no appreciable effect on the responsiveness of splenocytes to mitogens. The observed effect of RLX on cell-mediated and humoral immune responses does not appear to be the result of general toxicity.


Human recombinant interleukin-1 receptor antagonist (hrIL-1ra) enhances the stimulatory effect of interleukin-2 on natural killer cell activity against MOLT-4 target cells.

Interleukin-1 (IL-1) is considered an enhancer of host defence against malignancies. Patients with different diseases, including cancer patients with large tumour burdens, have demonstrated a reduced production of IL-1 from circulating leukocytes, in vitro. There are many naturally occurring substances which inhibit IL-1 activity aspecifically. Recently an interleukin-1 receptor antagonist (IL-1ra) has been discovered, which is secreted by human macrophages and is structurally similar to IL-1 beta (26% homology). The pretreatment of human peripheral blood mononuclear cells (PBMC) with hrIL-1ra (0.25-250 ng/ml) inhibits IL-1 alpha or IL-1 beta and enhances, in a dose-dependent manner, the stimulatory effect of IL-2 on their natural killer (NK) activity against a lymphoid cell line MOLT-4. The enhancing effect of IL-1ra on IL-2 activity was similar to that provoked by IL-1 beta. However, when IL-1ra was used alone without IL-2, no stimulatory effect was found compared with the control. In our data we show that a member of the IL-1 family, IL-1ra, has a significant effect on IL-2-stimulated NK activity against the MOLT-4 cell line. These studies provide new evidence of the biological potential of IL-1ra since this new protein enhances IL-2 activity on NK cells.


Hydrolysis and association of leucine enkephalin to lymphomic and erythroleukaemic cell lines--III. Soluble enzymes.

The possible existence of soluble proteolytic enzymes released by cells of lymphomic (U937 and 1301) and erythroleukaemic (K562) lines was studied measuring the hydrolysis of 3H-leucine enkephalin in the presence of cell-free supernatants. Results obtained indicate that in the presence of these supernatants leu-enkephalin rapidly disappears and that enkephalin disappearance is paralleled by the formation of peptides that can be interpreted as its hydrolysis fragments. Chromatographic analyses of cell supernatants indicate the presence of three groups of proteins active in leu-enkephalin degradation: aminopeptidases, dipeptidylaminopeptidases and dipeptidylcarboxypeptidases. In all three lines, soluble enzymes are represented by a sizeable number (from 13 to 25) of distinct activities. The number of enzymes identified and their considerable total activity suggest a possible role in the regulatory degradation of informational peptides, as proposed by several groups for the membrane-bound proteolytic enzymes of immunocompetent cells.


The distribution of anaerobic energy in 1000 and 4000 metre cycling bouts.

In sprinting events of short duration, performance depends not only on the mean external power output and the frictional losses but also on the distribution of energy over the race. To investigate the optimal distribution of anaerobic energy during cycling the 1000 m time trial and the 4000 m pursuit, we simulated a power equation which contains expressions for the production of aerobic and anaerobic power, for frictional losses and for the rate of change of kinetic energy. Parameters for air- and rolling resistance were derived from the literature and the equations for aerobic and anaerobic power production were based on supra-maximal bicycle ergometer tests. Simulated lap and final times were compared to those realized by the best four athletes during the 1990 World Championships. The mean final times of these athletes of 64.1 s and 272.6 s for the 1000 m and 4000 m respectively were closely approximated by the simulated times of 63.7 s and 281.3 s. The simulations show that performance in the 1000 m race depends to a great extent on a large power output at the onset of the race. Moreover, it is demonstrated that this distance should be cycled in an all out fashion and not with a uniform velocity after the start despite the higher air frictional losses. For the 4000 m pursuit it appears to be more effective to perform a short but powerful start and then continue the race with a constant or only slightly decreasing power output.


Acute exercise and immune function. Relationship between lymphocyte activity and changes in subset counts.

Twenty-one young male subjects exercised on a cycle ergometer for 60 min at 60% of VO2max. Blood samples collected every 30 min throughout exercise and continuing to 120 min recovery served for the immunological tests. Exercise induced biphasic changes in the various leucocyte subsets. There was a granulocytosis, lymphocytosis and monocytosis during exercise, and a further granulocytosis and a slight monocytosis, but a lymphocytopenia during recovery. All lymphocyte subsets (CD3+, CD19+, CD4+, CD8+, and CD16+ cells) increased in number during exercise, were decreased 30 min after exercise, and had not returned to baseline levels by 120 min of recovery. The apparent lymphocyte responsiveness to the mitogens phytohaemagglutinin (PHA) and pokeweed mitogen (PWM) declined significantly during exercise, returning to normal by 120 min of recovery. The natural killer (NK) activity rose markedly during exercise, but decreased to almost half the pre-exercise level at 30 and 60 min of recovery, returning to baseline levels after 120 min of recovery. Functional capability correlated well with the percentage of each major responder subset in the assay, suggesting that the in vitro lymphocyte PHA- and PWM-responsiveness and the NK activity did not change significantly on a per cell basis. The analysis of lymphocyte marker antigen density revealed that the CD3+, CD4+, CD8+ and CD19+ lymphocytes mobilized into the circulation during exercise did not express the respective CD3, CD4, CD8 and CD19 molecules as strongly as did the subsets circulating at rest, whereas the expression of the CD16 antigen on CD16+ lymphocytes remained unchanged.


A comparative study of blood lactate analytic methods.

Three different blood lactate analytic methods were tested for precision, accuracy, linearity, and intermethod comparison: a photoenzymatic assay (PHE), and three electroenzymatic (EE) semiautomatic assays (EE1, EE2, EE3). Reference standards and duplicate capillary blood samples from the earlobe were used. Precision and accuracy of the three techniques, when measuring L-lactate standards, were good in the whole range of measurement (mean variation coefficient, VC = 1.78-3.38%; mean difference = 1.81-3.38%). Correlation between the three methods was high (r = 0.913-0.946), but all three electroenzymatic techniques systematically measured lower values as compared to the PHE tests. The differences ranged from 0.1-1.2 (5 mmol.l-1 PHE level), to 3.4-5.7 (20 mmol.l-1 PHE level). These differences were drastically reduced when a hemolyser and a glycolytic inhibitor were added to the sample prior to the assay. The measurements obtained in capillary blood by the three techniques are not equivalent. The differences are partially attributed to the fact that the PHE technique measures total blood lactate, while the EE methods only measure plasmatic-extraerythrocytic lactate. Some regression equations are presented that may be used to convert values measured by the PHE technique, to EE values and vice versa.


Day to day variation in time trial cycling performance.

In an attempt to assess the reproducibility of laboratory cycling performance, eight well-trained (VO2max = 4.6 +/- 0.2 l.min-1) male cyclists completed 12 trials involving 4 successive performance rides at each of three total work outputs (approximately 1600, 200, and 14 kilojoules, respectively). These trials, designated as long, medium, and short trials (LT, MT, ST), represented exercise bouts of 105.12 +/- 0.41, 12.03 +/- 0.17 and 0.55 +/- 0.11 minutes, respectively. The trials, conducted on a computerized cycle ergometer in an isokinetic mode, were separated by a minimum of 72 hrs. All trials for each subject were completed at the same time of day. In all trials, subjects were allowed to select the pace in order to complete the ride in the shortest possible time. The mean coefficient of variation (CV) for performance time in each trial was: LT = +/- 1.01%, MT = +/- 0.95%, and ST = +/- 2.43%, respectively. The CV for performance time in ST was significantly greater than the CV in either LT or MT. In LT, performance time was significantly faster, and the mean % VO2max was significantly higher in trial 4 versus trials 1-3. There was no order effect in the MT or ST rides. The CV for mean VO2 (l.min-1), mean % VO2max, and RER during the LT rides were +/- 3.02%, +/- 3.64%, and +/- 3.53%, respectively. These data suggest that trained cyclists have the ability to reproduce endurance performance with a CV of approximately 1.0% in a time-trial protocol.(ABSTRACT TRUNCATED AT 250 WORDS)


Relationship between post-exercise plasma CK elevation and muscle mass involved in the exercise.

The purpose of this study was to investigate whether a larger post-exercise increase in plasma creatine kinase (CK) activity would be produced when a larger amount of muscle is damaged by eccentric exercise. Twenty-two non-weight trained females were placed into two groups; Group A (n = 12) and Group B (n = 10). Both groups performed 24 maximal eccentric actions of the forearm flexors on each bout. The right and left arm were exercised on the same day for Group A (24 eccentric actions per arm, a total of 48 actions). In contrast, Group B performed 24 actions with either the right or the left arm on the first bout and performed 24 eccentric actions with the opposite arm on the second bout 3-5 weeks later. Blood samples were taken before and for 5 days after each exercise and plasma CK activity was determined. Forearm flexion isometric force (ISO), range of motion evaluated by relaxed elbow joint angle (RANG) and flexed elbow joint angle (FANG), and perceived muscle soreness (SOR) were also examined to indirectly assess muscle damage. All of the muscle damage indicators changed significantly over time (p < 0.01) for both groups, but changes were not significantly different between arms or between groups. Because it seemed that both arms were equally "damaged" for each group, it was expected that Group A (two arms were exercised on the same day) should show an approximate two-fold increase in plasma CK compared to Group B when one arm was exercised on each bout.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of voluntary exercise on bone growth and calcium metabolism in spontaneously hypertensive rats.

The effects of voluntary exercise on bone length and calcium metabolism were investigated in spontaneously hypertensive rats (SHR) and Wistar rats. They were classified into voluntary exercise or sedentary group. Systolic blood pressure (SBP), bone length, urinary calcium excretion, plasma calcium and parathyroid hormone levels (PTH) were measured for analysis. SBP of the trained SHR (T-SHR) was significantly lower than that of the nontrained SHR (NT-SHR) at 10 and 11 weeks of age, but the trained Wistar rats (T-WR) and the nontrained Wistar rats (NT-WR) had similar SBP throughout the experimental period. The T-SHR had significantly longer bones in the extremities and the trunk than the NT-SHR, whereas the T-WR had similar or shorter bones than the NT-WR. The PTH level at 12 weeks of age was significantly lower in the T-SHR than in the NT-SHR. The urinary calcium excretion of the T-SHR was greater than that of the NT-SHR, but the food intake (calcium intake) of the T-SHR was also greater. The urinary calcium excretion and food intake of the T-WR and NT-WR were similar. No noticeable differences were observed in the plasma calcium level between the T-SHR and NT-SHR, the T-WR and NT-WR. From these results it is suggested that the abnormal calcium metabolism in the SHR is improved in some degree by voluntary exercise.


L-tryptophan supplementation does not improve running performance.

In 1988 Segura and Ventura (14) reported that 1.2 g of L-Tryptophan (L-TRY) supplementation increased total exercise time by 49.4% when the subjects were running at 80% of maximal oxygen uptake (VO2max). In human performance research, acute improvements of that category are rather uncommon. Both for this reason and because ingestion of purified L-TRY may have adverse effects, it seemed pertinent to repeat the investigation of Segura and Ventura. Forty-nine well-trained male runners, aged 18-44, with an average maximal aerobic power of 66 (57-78) ml.kg-1.min-1, participated in a randomized double blind placebo (P) study. Each subject underwent four trials on the treadmill. The first two served as learning experience, including measurement of VO2max and anaerobic threshold. During the last two trials the subjects ran until exhaustion at a speed corresponding to 100% of their VO2max-first an initial trial and then after receiving a total of 1.2 g L-TRY or P over a 24 hour period prior to the run. No significant difference between the improvements in the L-TRY and P group could be demonstrated. It is concluded that oral L-TRY supplementation does not enhance running performance.


Influence of acupuncture on physical performance capacity and haemodynamic parameters.

In a single blind study the question as to whether the needling of specific acupuncture points is able to produce an increase in physical performance capacity and better regulation of heart rate and blood pressure was examined. Thirty-six healthy young men were assigned at random to three groups, receiving either actual acupuncture, placebo acupuncture or no stimulation. Performance was determined by means of a spiro-ergometer test which was carried out at the beginning and at the end of five weeks of treatment consisting of one session per week. The subjects from the group which actually received acupuncture were able to increase maximum performance capacity significantly and also physical performance at the anaerobic threshold. This may be interpreted as a sign of functional improvement in haemodynamic and metabolic mechanisms. There was, on the whole, no noticeable effect produced by the placebo acupuncture. The control group, which received no stimulation, showed unfavourable changes in the values obtained compared with the results of the performance test at the commencement of the study.


Physical inactivity and depression in the community. Evidence from the Upper Bavarian Field Study.

In the Upper Bavarian Field Study a representative community sample of 1,536 persons (from 15 years of age and upwards) was interviewed by research psychiatrists. Twenty-seven percent of the respondents reported taking regular and 26.2% occasional physical exercise. Physical inactivity as well as depression were significantly associated with the female gender, increasing age, low socioeconomic status and the presence of a somatic disorder. Using a logistic regression model, the relation between physical activity and depression was measured by the odds ratio with subjects reporting regular physical activity as the reference group. Cross-sectional analysis revealed that after controlling for potential confounder variables the odds ratio for depression was, at 3.15, significantly higher for the physically inactive compared to the regular exercisers. The odds ratio for the group practising occasionally (1.55) was also elevated but not statistically significant, 87.3% of the subjects who participated in the baseline study were reinterviewed five years later. In contrast to the cross-sectional findings, low physical activity at wave 1 was not a risk factor for developing depression at wave 2.


Psychological effects during reduced training volume and intensity in distance runners.

Improved mood state has ben linked with reduced training (RT). However, RT has been measured only as a reduction in training volume, either maintaining or not considering the intensity of training employed. To investigate the effects of a 4-week reduction in both training volume and intensity on running performance and mood state, 10 well-trained adult male runners trained for 4 weeks at baseline training distance and pace (BT), followed by 4 weeks training reduced in volume by 66%, with intensity diminished so that all workouts were below 70% VO2max. Subjects completed the Profile of Mood States (POMS) before BT (PREBT), before RT (PRERT), and after RT (POSRT). They ran 5 km time trials PRERT and POSRT. Comparisons were made for the positive mood state of Vigor with an average of the values for the 5 negative mood states (NM) of the POMS. Eight of 10 changes from PRERT to POSRT were toward more negative mood. Nine of 10 runners required more time for the 5 km run POSRT than PRERT. Overall, 7 of 10 runners exhibited both an increase in 5 km time and a change toward more negative mood state (p < .004). However, the magnitudes of these changes were unrelated. These results suggest that mood state and running performance may be linked. Moreover, because mood state did not improve, this study suggests that studies dealing with the topic of reduced training should be specific with regard to the influence of both training intensity and volume.


Contextual issues for vocational placement coordinators: some preliminary findings.

While vocational placement is seen as an important component of the rehabilitation process, the experience of being a vocational placement officer has not been well documented in New Zealand. In the present study, previously developed and validated measures of occupational stress (defined as responsibility pressure, quality concern, role conflict, and job vs non-job conflict) and general life satisfaction were completed by 16 vocational placement officers employed by the Accident Compensation Corporation. Subjects also completed two open-ended questions assessing significant stresses in their work and strategies whereby stress could be reduced. There were high negative correlations between job stress and life satisfaction, and subjects reported particularly high levels of role conflict in their work. Open-ended stresses identified by subjects included: time pressure/workload problems; difficulties with negative societal and employer attitudes to disability; problems with clients, and; personal motivation problems. Most strategies for stress reduction that were identified were concerned with creating more effective communication and decision making processes in the organizational environment. Problems and limitations of the present study are addressed and future research directions are identified.


Informal caring networks among Chinese elderly with disabilities in Hong Kong.

Community care has been a major guiding policy in developing services for the elderly in Hong Kong. It has been assumed that Chinese families tend to have a large caring network to render care in the community. However, in a survey conducted in early 1991, it was found that such caring networks for disabled elderly people were very small and inconsequential. Quite a high proportion of Chinese disabled elderly are living alone in the community, and have a small network for emotional support. Effective measures to promote family care of disabled elderly people are discussed. Two pathways for integrating the formal and informal care sectors are examined in relation to promoting effective practice for the frail elderly living in the community.


Economic status of families living with multiple sclerosis.

A nation-wide survey of 604 families living with multiple sclerosis was conducted. The sample was recruited through local chapters of the National Multiple Sclerosis Society and the public press. This paper describes the economic impact of the disease on families. There were 427 families in which the woman had multiple sclerosis and 177 families in which the man had the disease. The average duration of the disease was 10 years. Family income was well above the national median income. Thirty-nine per cent of the men and 19% of the women with multiple sclerosis retired because of disability. Income was inadequate to pay for medical expenses in 21% of the families and 25% had inadequate funds to meet basic living expenses. While 80% of the families had health care insurance, for 28% of the families the insurance was inadequate to cover the cost of their illness. The number of children living at home and the amount of health care bills paid out-of-pocket failed to explain why families in the high income group had difficulty meeting their expenses. Factors other than income and proportion of medical expenses covered by insurance determine the economic impact on the family. If health care providers are going to meet all the needs of a family living with multiple sclerosis, a more comprehensive assessment of their economic status is needed.


Young blind children: towards assessment for rehabilitation.

Traditional assumptions about assessment need to be rethought, recast or rejected when blind children are to be assessed. Given that blind children follow a unique developmental route, rehabilitation specialists must take account of the impact of blindness on children and think of them also as clinically a special population. Dependence on standardized tests for the primary information about any child is always suspect; with regard to a blind child it is irresponsible. The authors describe the characteristics of an approach that is both systemic and common-sense. They illustrate it by reference to a new procedure (The Simmons-Davidson Developmental Profile) and to a case study.


Creating a disability mythology.

People with disabilities have, for the most part, failed to identify with each other as a group. This has been detrimental because it has built a sense of isolation when a camaraderie based upon existing commonalities could have been developed. During the past ten to twenty years, there has been a great deal of discussion about appropriate language to use when discussing disability issues. This discussion has been a part of a larger debate concerning the existence of a disability culture. I believe that there is indeed a disability culture and I am a proponent of identifying and passing on stories which contribute to that culture. I have chosen to use mythology to convey this message and have begun with a focus on heroes - people who do something out of the ordinary. It is contended that almost all people with disabilities have performed heroic activities because of the pervasive discrimination encountered by each individual with a disability. Creating a disability mythology is an attempt to recognize and promote heroes within the disabled community and to advocate the importance of telling other people how positive change has occurred through instances of individual heroism.


T-cell-receptor isoforms.

Early work on T-cell hybridomas lacking the T-cell-receptor (TCR) sub-unit CD3 eta had suggested a correlation between the presence of CD3 zeta-eta heterodimers and signalling leading to phosphatidyl-inositol (PI) turnover as well as activation-induced cell death. The cloning of CD3 eta has now allowed thorough and direct analysis of the signal transduction properties of CD3 zeta-zeta-, CD3 zeta-eta- and CD3 eta-eta-containing TCRs. We have found that all forms of the TCR are capable of transducing signals leading to PI turnover, Ca2+ mobilization, IL-2 production and cell-cycle arrest. CD3 zeta and CD3 eta utilize the same promoter which yields coordinate expression of both products, so that restricted CD3 eta expression in a sub-population of thymocytes is unlikely. Immunohistochemical methods employing an anti-CD3 eta-specific monoclonal antibody (MAb) show no detectable staining of thymic sections from adult mice, implying at best a low level of constitutive CD3 eta expression. In contrast, CD3 eta expression is readily detected in the majority of cortical thymocytes of CD3 eta transgenic mice using a Thy-1 promoter construct. However, over-expression of CD3 eta in mice transgenic for this polypeptide does not result in increased negative selection in vivo, consistent with the in vitro findings that induction of cell death is not strictly dependent on CD3 eta. Despite earlier reports of the detection of human CD3 eta protein, we find no CD3 eta message in human thymus or T cells. Cloning of the human CD zeta-eta genomic locus has demonstrated approximately 70% homology between the mouse and human genomic sequence, corresponding to the mouse CD3 eta-specific exon. However, translation of the DNA sequence does not result in a homologous amino acid sequence. Thus, there does not appear to be a CD3 eta protein in humans.


Structure and function of the CD16:zeta:gamma complex expressed on human natural-killer cells.

Natural-killer (NK) cells are large granular lymphocytes involved in host defense against tumor cells and virally-infected cells. In addition to natural cytotoxicity, NK cells can effect antibody-dependent cytotoxicity mediated by CD16 (Fc gamma RIIIA alpha). It has recently been shown that CD16 is associated with disulfide-linked dimers composed of 2 homologous sub-units, zeta and gamma. These transducing molecules are also associated with other multimeric cell-surface receptors such as the T-cell antigen receptor (CD3:TCR) complex (zeta and gamma) and the high-affinity Fc receptor for IgE (Fc epsilon RI) expressed on basophils and mast cells (gamma). These results show that distinct cell-surface receptors utilize common transducing sub-units, and emphasize the homology between the CD16, Fc epsilon RI and CD3:TCR complexes. However within the lymphoid cells, the gamma-gamma homodimer is preferentially expressed in NK cells and cytotoxic T cells, suggesting that specific combinations of these transducing dimers might sub-serve distinct signal-transducing functions, and contribute to lymphocyte heterogeneity.


Targeting of T or NK lymphocytes against tumor cells by bispecific monoclonal antibodies: role of different triggering molecules.

MAbs directed against triggering surface molecules expressed by T lymphocytes (CD3, TCR, CD2, CD28) or by NK cells (CD2, CD16) are able to induce the functional program of these cells. These MAbs represent suitable reagents to construct biMAbs directed against TAA, in order to specifically target effector lymphocytes against tumor cells. Anti-CD3/anti-EGF-R biMAbs were constructed to specifically direct T lymphocytes against EGF-R+ tumor cells. Such biMAb are able to induce cytolysis of EGF-R+ tumor cell lines (A431, IGROV, KATO-III and U-87) by cytolytic CD3+ effector lymphocytes while tumor cells having low or absent expression of EGF-R were not lysed. In addition, both cytolytic T (CD8+) cells and non-cytolytic (CD4+) IL-2-expanded lymphocytes were able to secrete lymphokines upon contact with EGF-R+ tumor cells. To target NK cells against NK resistant ovarian carcinomas, we used an anti-CD16 Mab (IgG1) together with an anti-ovarian carcinoma MAb (IgG2a), to construct biMAbs using the hybrid hybridoma technique. The hybrid IgG1/IgG2a biMAb triggered the specific lysis of relevant target cells by resting NK cells and by a subset of NK clones. In addition, some TCR gamma/delta+ clones but not TCR alpha/beta+ clones could be targeted by the biMAb.


Mechanisms of lymphocyte traffic and cell targeting.

Several concepts in the immunotherapy of cancer aim to exploit the powerful cellular effector mechanisms of the immune system. In contrast to antibodies and other soluble agents, cells do not distribute randomly with the circulation but have their own program of trafficking through the body and entering the various organs. Some characteristics of migration and homing of different populations of lymphocytes as well as the molecular mechanisms governing lymphocyte entry into tissues are discussed here in order to delineate problems and prospects in targeting cell populations into specific organs.


Targeted cytokine production.

It has been well established that bispecific antibodies containing anti-T-cell receptor MAbs crosslinked to anti-tumor MAbs induce T cells to lyse tumor cells, as measured in a 51Cr-release assay. Such lysis requires direct attachment between target and cytotoxic cells and most probably involves the exocytosis of cytolytic substances into the cell:cell interface. In addition, targeted T cells mediate a second activity, the secretion into the medium of factors that can block the growth of bound tumor cells and unbound bystander cells. In order to test how targeted effector cells mediate anti-tumor effects in vivo, we are currently developing a totally syngeneic murine system in which murine T cells are targeted against mouse mammary tumors. The system allows us to treat both primary tumors and tumor transplants, using a mammary-tumor-virus antigen as the entity that is specifically recognized on the tumor cells.


Activation of T cells in vivo using anti-CD3 and staphylococcal enterotoxins.

One of the major problems in clinical immunity is that neoplastic and virally infected cells are insufficiently immunogenic to trigger an immune response. During the past several years, our laboratory has explored the use of T-cell-specific reagents including monoclonal antibodies (MAbs) and staphylococcal enterotoxins to amplify immune responses. This report summarizes our efforts to augment immunity with these agents.


Possible targets on carcinoma for bMAb retargeting of lymphocyte or drug cytotoxicity.

Retargeting of drugs or lymphocyte cytotoxicity through bi-specific monoclonal antibodies (bMAbs) has been proven a therapeutic tool against human carcinoma both in pre-clinical in vitro and in vivo studies. Some of these reagents have already been introduced into clinical trials and preliminary results appear to be promising. However, improvement of the specificity of this approach could be achieved by selecting more suitable target molecules on tumor cells. The research focused on developing MAbs directed against molecules with tumor-restricted distribution and homogeneous expression. Cell-membrane receptors for nutrients or growth factors which operationally represent tumor-specific molecules due to their overexpression, could be considered appropriate targets. Several bMAb anti-nutrients (the folate-binding protein) or growth-factor receptor (c-erbB1, c-erbB2)/anti-triggering molecules have been generated and all were able to efficiently retarget the relevant population of lymphocytes on tumor cells. In order to design a more reliable and selective therapeutic tool, the following parameters were analyzed: correlation between cytotoxicity and antigen level, potency and possible modulation of the target molecule.


Engineering a humanized bispecific F(ab')2 fragment for improved binding to T cells.

We recently constructed a humanized bispecific antibody (BsF(ab')2v1) by separate E. coli expression of each Fab' arm followed by directed chemical coupling in vitro. BsF(ab')2 v1 (anti-CD3/anti-p185HER2) was demonstrated to retarget the cytotoxic activity of human CD3+ CTL in vitro against the human breast-tumor cell line, SK-BR-3, which over-expresses the p185HER2 product of the proto-oncogene HER2. Our minimalistic humanization strategy is to install as few murine residues as possible into a human antibody in order to recruit antigen-binding affinity and biological properties comparable to that of the murine parent antibody. This strategy proved very successful for the anti-p185HER2 arm of BsF(ab')2 v1. In contrast BsF(ab')2 v1 binds to T cells via its anti-CD3 arm much less efficiently than does the chimeric BsF(ab')2 which contains the variable domains of the murine parent anti-CD3 antibody. Here we have constructed additional BsF(ab')2 fragments containing variant anti-CD3 arms with selected amino acid replacements in an attempt to improve antibody binding to T cells. One such variant, BsF(ab')2 v9, was created by replacing 6 residues in the second hypervariable loop of the anti-CD3 heavy chain variable domain of BsF(ab')2 v1 with their counterparts from the murine parent anti-CD3 antibody. BsF(ab')2 v9 binds to T cells (Jurkat) much more efficiently than does BsF(ab')2 v1 and almost as efficiently as the chimeric BsF(ab')2. This improvement in the efficiency of T-cell binding of the humanized BsF(ab')2 is an important step in its development as a potential therapeutic agent for the treatment of p185HER2 over-expressing cancers.


Janusin: new molecular design for bispecific reagents.

It is well established that soluble CD4 (sCD4) inhibits HIV infection in vitro, regardless of the virus strain or genetic variant. Most effective molecules, thus far, based on sCD4 are those in which CD4 is combined with immunoglobulin constant regions (CD4-IgG or CD4-IgM). Such molecules maintained HIV-gp120 specificity mediated by CD4 and also antibody effector functions such as complement activation, Fc receptor binding, long serum half-life or transport across the placental barrier. We have now developed sCD4 molecules which are even more potent anti-HIV reagents. These molecules are based on the principle of bispecific antibodies and they have properties capable of retargeting cytotoxic T lymphocytes onto HIV-infected cells and inducing efficient killing. CD4 combined with anti-human CD3 (FvCD3) single-chain combining site has been produced (CD4-FvCD3-JANUSIN). This molecule shows the expected biological activities, namely, binding to the 2 ligands, human CD3 and gp120, also efficiently retargeting CTLs of any specificity onto HIV-infected cells. In addition, several advantages over classical bispecific antibodies can be achieved: only one polypeptide, not a mixture containing the desired product, is produced, thus simplifying the purification process. In addition, Janusin designs do not contain the Ig Fc portion, which could mediate illegitimate retargeting of T-cells. In addition to CD4-FvCD3-JANUSIN, receptor-Fv, Fv-Fv or ligand-Fv Janusins can be produced.


Functional assembly of chimeric T-cell receptor chains.

We have generated cytotoxic T-cell hybridomas expressing chimeric T-cell receptors (cTCR) with an antibody-type specificity for the TNP hapten. Transfectants expressing the cTCR genes could mediate specific lysis of haptenated tumor cell lines of various types and secrete IL-2 upon stimulation with TNP modified cells. In a previous report, we showed that double-gene transfectants expressing either VHC alpha and VLC beta or VHC beta and VLC alpha could be activated by TNP-modified stimulator cells or TNP proteins immobilized on plastic. Single-chain transfectants (expressing VHC alpha or VHC beta alone) could be mainly activated by TNP-cells. We now report that transfection of chimeric VHC alpha gene into an alpha-chain-defective mutant restores the surface expression of the TCR/CD3 complex. In parallel, such transfectants regained the ability to respond to mitogen and anti-CD3 antibodies and responded weakly to TNP cells. Double gene transfectants, bearing 2 complementary chimeric chains, expressed high amounts of cTCR on their surface, sufficient to acquire sound anti-TNP reactivity. Cells expressing the VHC beta gene only were not functional and had no detectable surface TCR chains. Taken together, our results suggest that chimeric VHC alpha chains can pair with endogenous V beta C beta chains, but that there is preferential association between complementary chimeric chains, resulting in higher functional expression of the chimeric TCR.


Bispecific-antibody-mediated targeting of radiolabeled bivalent haptens: theoretical, experimental and clinical results.

Chemically conjugated bispecific (anti-cell surface antigen, anti-hapten) Fab'-Fab antibodies (Bs-MAbs) have been used to target 125I-, 111In- and 99mTc-labeled haptens to cell sub-sets. In vitro, bivalent haptens were found to bind more strongly than their monovalent analogs to the Bs-MAbs bound to ("ordered" on) the cell surface, or than to free ("disordered") Bs-MAbs: they are selective for cell-bound Bs-MAbs. In tumor-grafted nude mice models, the sequential injections of microgram amounts of Bs-MAb, and 1 day later, of microC amounts of bivalent haptens permits to sharply delineate small tumors (using a gamma camera), hours after injection. Further, the isotope biodistribution was found to be at least 3 times more selective for the tumor than that obtained with directly labeled anti-CEA F(ab)'2 or with monovalent haptens. This better in vivo selectivity of the 2-step targeting of bivalent haptens was also demonstrated in a pharmacokinetic study using therapeutic amounts of reagents. In primary-colon-carcinoma patients, a similar comparative immunoscintigraphy study confirmed the better selectivity of bivalent hapten targeting over direct targeting, on the basis of image quality and ex vivo tissue counting. In patients with medullary carcinoma of the thyroid, bivalent hapten targeting allowed us to confirm tumor extension and to find occult lesions. Interestingly, radio-immunoguided surgery was necessary to resect these small lesions. These experimental results, together with technological and theoretical considerations, suggest that Bs-MAb-mediated targeting of isotopes (or other agents) is one of the major ways to increase the clinical performance of MAb-based targeting diagnostic and therapeutic tools.


Novel surface molecules involved in human NK cell activation and triggering of the lytic machinery.

Three new monoclonal antibodies (MAbs) termed 7A6, PP35 and A6/143 were isolated after mouse immunization with CD3- CD16+ NK clones. The screening procedure was based on the ability of MAbs to trigger cytolytic activity of the immunizing clones in a re-directed killing assay against the P815 murine mastocytoma cell line. The 7A6 MAb reacts with 58 kDa surface molecules that appear to belong to the same molecular family defined by the previously described NK-sub-set-specific GL183 and EB6 MAbs. However, unlike from these MAbs, the 7A6 MAb reacted with (and activated) all CD3- NK lymphocytes, independent of their sub-set assignment (based on the expression or lack of expression of EB6, GL183 and CD16). The PP35 MAb reacted with a 70 kDa surface molecule expressed on all CD3- NK cells, as well as on TCR gamma/delta + cells and on a small sub-set of TCR alpha/beta + CD8+ lymphocytes. The PP35 MAb induced activation of essentially all NK cells, although clonal analysis revealed quantitative differences in the magnitude of the cytolytic responses elicited in different clones. Finally, the A6/143 MAb reacted with a molecule of 115 kDa expressed by all human PBL. Similarly to 7A6 and PP35 MAbs, the A6/143 MAb activated all sub-sets of cloned NK cells.