Electron spin resonance determination and superoxide dismutase activity in polymorphonuclear leukocytes in congestive heart failure. Electron spin resonance spin trapping technique was used to measure the generation of active oxygen free radicals during the respiratory burst of phorbol myristate acetate-stimulated leukocytes, and the superoxide dismutase activity in healthy subjects and in patients with congestive heart failure. The authors also measured the concentration of peroxidation products (primarily malondialdehyde) by the thiobarbituric acid method. Experimental results showed that the electron spin resonance spectra obtained during the respiratory burst of polymorphonuclear leukocytes stimulated with phorbol myristate acetate primarily were those of the spin adduct of 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) with the superoxide anion and to a lesser extent those of DMPO with hydroxyl radical. Compared with healthy subjects, the release of oxygen free radicals in the respiratory burst of polymorphonuclear leukocytes stimulated with phorbol myristate acetate and the concentration of thiobarbituric acid reactive product in plasma were significantly increased in patients with congestive heart failure while the activity of superoxide dismutase was markedly lower. The increased production of oxygen free radicals by polymorphonuclear leukocytes and the decreased capability of antioxidative defences might play an important role in the generation and development of cardiac failure.


Arginine induced insulin release in patients with newly onset non-insulin-dependent diabetes mellitus. To examine the release of insulin in response to oral glucose, intravenous glucagon and intravenous arginine, we measured the levels of plasma glucose, immuno-reactive insulin (IRI) and C-peptide levels on fasting and following an oral glucose loading (OGTT), intravenous glucagon (GON) and arginine (ARG) infusion test in nine newly diagnosed non-insulin dependent diabetics. Their ages ranged from 38 to 65. The fasting plasma glucose and hemoglobin A1c levels were 240 +/- 14 mg/dl (Mean +/- SEM) and 10.7 +/- 0.54%, respectively. Mean values of the peak C-peptide/fasting C-peptide ratio and peak IRI/fasting IRI ratio were significantly increased, as compared with the basal level (P < 0.05), but not significantly different from those of the OGTT, GON and ARG test. In conclusion, the effect of arginine-induced insulin secretion in non-insulin dependent diabetes mellitus is as good as those of glucose or glucagon.


Measurement of separate glomerular filtration rate by means of 99mTc-DTPA. We detected the separate glomerular filtration rate (SGFR) with 99mTc-DTPA in patients on the nephrology ward since 1987. There were 40 patients, 19 males and 21 females. Their age ranged from 15 to 79 years old, with a mean +/- 1SD of 44 +/- 17 years old. They all received a SGFR examination and one or two creatinine clearance (Ccr) examinations during hospitalization. The Ccr values ranged from 4.6 to 141.5ml/min, and the mean +/- 1SD was 68.1 +/- 37.9ml/min. The total GFR value by the SGFR method was 1 to 142ml/min, with a mean of 78.9 +/- 39.4ml/min. The correlation coefficient of Ccr and SGFR was r = 0.81 (p = 0.0001). All 16 patients who had a SGFR less than 40% of the total GFR had specific clinical problems in that particular kidney. We concluded that 99mTc-DTPA renography was a correct and rapid method to check GFR, and may be another choice in addition to the traditional Ccr measurement. It was also a useful method for the diagnosis of disease in one kidney.


Operative laparoscopy in benign cystic teratoma of ovary. Eighteen women of reproductive age underwent laparoscopic management of unilateral ovarian benign cystic teratoma from 3 to 7 cm in diameter. Nine patients received pelviscopic enucleation of ovarian cysts and nine patients received unilateral oophorectomy or salpingo-oophorectomy. No major surgical complication was encountered and the hospital stay was short. Our series suggest that operative laparoscopy is a safe and effective method in the management of benign cystic teratoma.


Clinical application of ejection fraction by gated MRI. To evaluate the clinical desired accuracy of the left ventricular ejection fractions (EFs) calculated by magnetic resonance imaging (MRI). Within one to three days post coronary angiography and left ventriculogram, twenty-five adults were studied by MR imaging. They had nuclear medicine studies for left ventricular ejection fractions as well. EKG-gated spin-echo 30-msec echo-delay images were obtained in end systole and end diastole in a plane parallel to the ventricular septum. Analysis of ventricular volumes and ejection fractions were performed using the area-length method. The EFs calculation by gated MRI were compared with that obtained by angiocardiography and nuclear medicine studies. The linear regression line obtained for ejection fraction was y = 0.858x +6.813, r = 0.753, p = 0.009; y' = 1.567x'-24.692, r' = 0.783, p' = 0.002. The above figure indicate a reasonable correlation among these three methods.


Distal paresthesia in patients with cervical cord lesions. Nine patients complaining of distal paresthesia were found to have spinal cord lesions with no evidence of peripheral nerve disease. This pattern may be due to central nervous system dysfunction, with a lesion being intramedullary or extramedullary. The sensory deficits were initially asymmetric. Paresthesias could originate from the hands and progress to the stocking-glove pattern. It was usually more severe in hands than in feet. The abnormal somatosensory evoked potentials, found in all patients, indicated a lesion involving the posterior column, a site supposed to be the pathologic basis of paresthesia. As hands and feet have relatively large somatotopic representation in the posterior column of the cervical cord, the paresthesia tends to localize over distal portions of extremities.


Pericardiocentesis: a 20 patients study. Emergency pericardiocentesis, guided by a two-dimensional echocardiography, was performed on twenty patients with symptomatic pericardial effusion of various types and causes. There were fourteen men and six women. The underlying causes were: primary lung cancer (6 cases), metastatic cardiac tumors (3 cases), tuberculosis (4 cases), complicated interventional procedures with cardiac chamber or vessel perforations (2 cases), dissecting aortic aneurysm (1 case), systemic lupus erythematous (1 case), idiopathic pericarditis (1 case), bacterial pericarditis (1 case), and myxedema heart disease (1 case). Seventeen cases were performed through the left xipho-sternal approach and 3 cases through the apical approach. None of the patients died as a result of these procedures. A two-dimensional echocardiogram is useful in diagnosing cardiac tamponade as well as in guiding pericardiocentesis, and obtaines highly positive results (20/20). The positive rate of pericardial fluid cytology for malignant cells was 89% (8/9), however, pericardial fluid cultures or direct smear for tuberculosis were negative (0/4). In cancer patients, the mean survival time following pericardiocentesis was 4.2 months (range, 1-7.8 months). We concluded that neoplastic involvement of the pericardium is the most frequent cause of symptomatic pericardial effusion. Pericardiocentesis assisted by a two-dimensional echocardiogram is safe and easy. In addition, pericarditis caused by TB is still significant and must be considered in every case in our nation.


Superior vena cava syndrome in children with malignancy: analysis of seven cases. Superior vena cava syndrome (SVCS) is rare in children. In Veterans General Hospital-Taipei, a total of 364 cases of SVC syndrome were diagnosed in the past 12 years. Of them only seven cases were younger than 18 years of age, ranging from 6 to 17 years, and they were all caused by mediastinal tumor. The underlying malignancy included malignant lymphoma in 4, teratocarcinoma in one and unknown in 2. The most common initial symptom was cough, followed in order of frequency by chest discomfort or neck mass. Dyspnea, orthopnea, swelling of head and neck, and venous engorgement might develop gradually within one to three weeks. Of the reported seven cases, two cases received immediate resuscitation upon arrival but expired in 1-2 hours. The other five cases received treatment with intravenous steroid as well as chemotherapy, and three cases also received committent emergent radiotherapy. Two of them expired 4 months and 2 years after treatment, respectively. Of the two surviving cases, one has received a complete course of chemotherapy and the other is still under regular chemotherapy in our hospital. Both of them are stable till now.


[Epidemiologic study of head injuries in Taipei City, Taiwan]. This study was designed to examine the descriptive epidemiology of head injuries in an urban population in the Taiwan area, Taipei City, during the period from July 1, 1987 to June 30, 1988. Clinical records reviewed included emergency room (ER) charts, inpatient charts of 19 major hospitals, death registration forms and medical examiners' reports in this city. A formulated definition was used to identify patients with head injuries. A total of 4,692 cases were collected, which included 4,319 hospital inpatients and 373 non-hospital deaths. The average incidence rate for head injury was 180/100,000 per year, 246/100,000 for males and 111/100,000 for females. The highest incidence rate was observed in the elderly group followed by the age group of 20-29 years. Sixty eight percent of the head injuries were caused by traffic accidents, 59.7 percent of which were involved in motorcycle rides. This rate was higher than those in any western reports. Among the 10-39 age group, the head injuries caused by traffic accidents were mainly due to motorcycle accidents. However, most of the children and the elderly who were injured were either pedestrians or bicycle riders. The average mortality rate was 23/100,000 per year, 34/100,000 for males and 12/100,000 for females. Initially the Glasgow Coma Scale was used in assessing the severity of head injuries. Seventy-two percent of the cases were considered mild, and 28 percent moderate to severe in degree, including 609 deaths.(ABSTRACT TRUNCATED AT 250 WORDS)


[Motor evoked potentials in patients with cervical spondylotic myelopathy]. We applied somatosensory and motor evoked potential studies to investigate patients with cervical spondylotic myelopathy (CSM) diagnosed clinically and roentgenographically. There were 32 consecutive cases collected, including 25 males and 7 females, at an average age of 62 +/- 8.2 (49-75) years. A control group consisted of 30 healthy subjects matched in body height and age with patients. The normal values of head-to-neck and head-to-lumbar region central motor conduction times (CMCTs) were 7.04 +/- 0.60 (6.1-8.2) and 15.94 +/- 1.23 (14.0-18.7) ms, which were not affected by body height. The somatosensory evoked potential (SEP) findings were related to clinical posterior column signs, levels of cord lesion, and nerves tested. The abnormality rate of median nerve SEP was relatively low, 53% (17/32), particularly for the lesion below C5-6 level in which median nerve SEP was usually normal. The abnormality rates of ulnar and tibial nerve SEPs were up to 84.5% (27/32) and 90.6% (29/32) respectively. However, median, ulnar and tibial nerve SEPs were normal in 3 (9.4%) patients showing normal posterior column function clinically. Only 18 (56.3%) patients had recordable motor evoked potential (MEP) from abductor digiti minimi (ADM) to head stimulation. The head-to-neck CMCT, 9.12 +/- 1.53 (6.3-12.7) ms, was significantly prolonged. Of these 18 cases there were 9 cases of delayed CMCT with reduced MEP amplitude, 4 of delayed CMCT, 3 of reduced MEP amplitude, and 2 of normal CMCT and MEP amplitude. The remaining 14 cases (43.7%) showed absent MEP from ADM to head stimulation. The overall abnormal rate of MEP from ADM was 93.8% (30/32). MEP from tibialis anterior (TA) to head stimulation was recorded only in 6 patients in whom the responses were all abnormal. There was no response recorded from TA in the remaining 26 CSM patients.(ABSTRACT TRUNCATED AT 250 WORDS)


[Detection of the myelination process of Chinese infants by 1.0T MRI]. To establish the myelination process of brain of Chinese infants, our study consisted of seventy-five infants ranging in age from 5 days to 2 years old. They were studied with a 1.0 T superconductive MRI. In the new born, myelination could be seen in the brainstem, middle cerebellar peduncle, and posterior limb of the internal capsule in both T1WI and T2WI. Then the myelination process rapidly progressed month by month. Myelination of the centrum semiovale and optic radiation could be seen in T1WI in the newborn up to 3 months old, and in T2WI at 3-5 months old. The myelination of the splenium and the genu of corpus callosum were seen at 3-6 months in T1WI and 5-6 months in T2WI. Myelination of the occipital lobe and frontal lobe were seen at 3-6 months and 5-10 months in T1WI, and 8-14 months, 10-16 months in T2WI respectively. In 7-10 months, the brain appeared as an "early adult pattern" in T1WI; while, in T2WI, the brain appeared as an "early adult pattern" at 14-18 months old. The understanding of the myelination process in infants is important and essential for the judgement of the infantile brain and for the diagnosis of many CNS diseases of infants.


[The pattern of fresh frozen plasma transfusion in Veterans General Hospital-Taichung]. Conducted a survey at Veterans General Hospital in Taichung, to compile a statistical analysis on the usage of fresh frozen plasma (FFP) transfusions. The investigation was conducted from July 1st to September 30, 1991. According to the distribution of usage of FFP, we investigated 726 transfusions (4,216 units) based on our grouping criteria. We found the following: 532 units were used for clotting support which accounts for 12.6% of total, 815 units (19.3%) for blood pressure support, 148 units (3.5%) for combination of clotting and blood pressure support, 681 units (16.2%) for albumin replacement, 436 units (10.3%) for therapeutic pheresis, 396 units (9.4%) for packed red cell concurrently, 819 units (19.4%) for burn cases, and the remaining (9.2%) for other or unidentified reasons. If it was classified by the departments applying FFP; 76% (3,200 units) were used by surgery, 22.4% (947 units) by internal medicine, and 1.6% (69 units) by other departments. Having classified all the reasons for FFP transfusion, we found that FFP was commonly used as a volume expander, for nutrition support, and reconstituted whole blood. These reasons are out of the range of indications for FFP transfusion. This misuse of FFP transfusion increases the chance of transmission infections therefore, we will thoroughly investigate these treatment modalities in order to ensure our blood source is being used in an appropriate manner. This will allow patients the best possible treatment available.


[Leukemia cutis: clinical and histopathological analysis of 14 cases]. We report 14 cases of leukemia cutis registered at Department of Dermatology, Veterans General Hospital Over a period of 18 years. There were one patient with acute lymphocytic leukemia (ALL), one with chronic lymphocytic leukemia (CLL), seven with acute monocytic leukemia (AMOL), one with acute myelomonocytic leukemia (AMML), and four with chronic myelocytic leukemia (CML). Multiple papules and nodules were the most frequent clinical lesions. Metastatic skin lesions occurred most commonly on legs (71%), followed by arms (64%), back (50%), anterior chest (50%), scalp (14%), and face (14%). The feet (7%) were rarely involved while palms and soles were rarely involved. Cutaneous leukemic lesions may be concomitant with or after, but never before the diagnosis of systemic leukemia in our series has had such change. In general, the histopathology of leukemia cutis showed diffuse or nodular infiltration of leukemic cells in the dermis and subcutaneous tissue, often typing of leukemia relays on more confirmative studies of peripheral smear and bone marrow biopsy. Leukemia cutis seems to be dissemination of systemic leukemia to the skin, and the presence of cutaneous leukemic lesions are associated with a very poor prognosis. Most patients (85%) died within 4 months after appearance of skin metastasis.


Rotator interval tear: a case report.

Rotator interval tear: a case report. Rotator cuff tear is one of the most common shoulder disorders. Most of the tears involve the critical area of the supraspinatus tendon. The rotator interval tear is extremely rare. Tear in this area is important due to its relationship with shoulder instability, different clinical symptom and different management. Recently we have uneventually treated a case with rotator interval tear after a violent throwing injury. The purpose of the paper is to emphasize the clinical differences of this unusual tear.


Fibromatosis of the chest wall in children: a case report. Fibromatosis is often found in the anterior abdominal wall of parous women. It occurs rarely in the chest wall and is rarely found in children. Our case was a 7-year-old male patient with a rapid-growing, painless tumor in the right anterior chest wall for 6 months. He received a resection of the tumor, the bilateral clavicular heads, the upper-third of the sternum and segments of the first and second ribs bilaterally. Reconstruction was conducted by transposition of the pectoralis major muscle and a prosthesis of wire mesh with methyl methacrylate. Histologically, the tumor showed dense collagenous tissue consistent with mature aggressive fibromatosis. It was a benign tumor without distant metastatic potential but with an aggressive clinical behavior. Although postoperative radiotherapy is usually indicated in adults, it was reserved due to the immature skeleton of this boy. In this case, the use of a rigid prosthesis avoided tracheostomy and postoperative ventilation.


Indomethacin worsens and a leukotriene biosynthesis inhibitor accelerates mucosal healing in rat colitis. The implication of leukotrienes as mediators of inflammation and recent evidence that prostaglandin analogues provide a beneficial effect during experimental colitis led to the speculation that (i) leukotrienes may be injurious and (ii) prostaglandins may be protective to colonic mucosa. Using a 2% acetic acid induced rat colitis model, we administered specific cyclooxygenase (indomethacin) and leukotriene biosynthesis inhibitors (MK-886) to examine the effect of endogenous prostaglandins and leukotrienes on colonic macroscopic injury, mucosal inflammation as measured by myeloperoxidase activity, net in vivo intestinal fluid absorption, and colonic PGE2 and LTB4 levels as measured by in vivo rectal dialysis. Indomethacin treatment prior to induction of colitis reduced endogenous mucosal PGE2 levels and exacerbated macroscopic ulceration and net fluid absorption. Addition of the exogenous PGE1 analogue misoprostol to the indomethacin-exacerbated colitis completely healed colonic macroscopic ulceration and inflammation but only partially improved fluid absorptive injury. The specific leukotriene biosynthesis inhibitor MK-886 administered prior to induction of colitis healed macroscopic ulceration and inflammation but not fluid absorptive injury. This mucosal reparative effect of MK-886 occurred at a dose that reduced colonic LTB4 synthesis while concomitantly enhancing PGE2 levels. Combining MK-886 with misoprostol treatment improved not only macroscopic ulceration and inflammation but also provided a synergistic effect that maintained net colonic fluid absorption at noncolitic control levels. These studies suggest that, during the induction of experimental colitis, endogenous prostaglandins play a pivotal role in providing a mucosal healing effect, and that leukotriene biosynthesis inhibitor may manifest part of its beneficial effect by shifting arachidonic acid metabolism towards production of prostaglandins.


Influence of arachidonic acid metabolites on cardiac alpha 1-adrenoceptor responses in diabetic rats. There is evidence that one or more metabolites of arachidonic acid can produce positive inotropic effects and may also be implicated in the enhanced alpha 1-adrenoceptor responses in hearts from diabetic rats. We therefore carried out a study to investigate the possibility that arachidonic acid metabolites could be involved in the altered cardiac alpha 1-effect of norepinephrine (in the presence of propranolol) in chronic streptozotocin-diabetic rats. Our results have shown that in the presence of the cyclooxygenase inhibitor indomethacin or the thromboxane synthetase inhibitor imidazole, the norepinephrine-stimulated positive inotropic effect and the formation of inositol 1,4,5-trisphosphate were significantly increased in control hearts but were unaltered in hearts from diabetic rats. The addition of the prostacyclin synthetase inhibitor tranylcypromine reduced the norepinephrine-stimulated positive inotropic effect and inositol 1,4,5-trisphosphate formation only in diabetic hearts and had no effect in the controls. The nature and physiological significance of the enhanced positive inotropic effect and inositol 1,4,5-trisphosphate formation in the control heart with the addition of indomethacin and imidazole are still unclear. The effect of tranylcypromine may indicate the participation of prostacyclin in mediating the enhanced alpha 1-inotropic effect of norepinephrine in the chronic diabetic heart.


Pharmacology of MK-0591 (3-[1-(4-chlorobenzyl)-3-(t-butylthio)-5-(quinolin-2-yl-methoxy)- indol-2-yl]-2,2-dimethyl propanoic acid), a potent, orally active leukotriene biosynthesis inhibitor. MK-0591 (3-[1-(4-chlorobenzyl)-3-(t-butylthio)-5-(quinolin-2-yl-methoxy)- indol-2-yl]-2,2-dimethyl propanoic acid, previously L-686,708) is a potent inhibitor of leukotriene (LT) biosynthesis in intact human and elicited rat polymorphonuclear leukocytes (PMNLs) (IC50 values 3.1 and 6.1 nM, respectively) and in human, squirrel monkey, and rat whole blood (IC50 values 510, 69, and 9 nM, respectively). MK-0591 had no effect on rat 5-lipoxygenase. MK-0591 has a high affinity for 5-lipoxygenase activating protein (FLAP) as evidenced by an IC50 value of 1.6 nM in a FLAP binding assay and inhibition of the photoaffinity labelling of FLAP by two different photoaffinity ligands. Inhibition of activation of 5-lipoxygenase was shown through inhibition of the translocation of the enzyme from the cytosol to the membrane in human PMNLs. MK-0591 was a potent inhibitor of LT biosynthesis in vivo, first, following ex vivo challenge of blood obtained from treated rats and squirrel monkeys, second, in a rat pleurisy model, and, third, as monitored by inhibition of the urinary excretion of LTE4 in antigen-challenged allergic sheep. Inhibition of antigen-induced bronchoconstriction by MK-0591 was observed in inbred rats pretreated with methysergide, Ascaris-challenged squirrel monkeys, and Ascaris-challenged sheep (early and late phase response). These results indicate that MK-0591 is a potent inhibitor of LT biosynthesis both in vitro and in vivo indicating that the compound will be suitable for assessing the role of leukotrienes in pathological situations.


Chronic low frequency stimulation reduces myosin phosphorylation in rabbit fast twitch muscle. The effect of 1-12 days of electrical stimulation (10 Hz) on the ability to phosphorylate the P-light chain of myosin was studied in rabbit tibialis anterior muscle. Myosin phosphorylation was induced by exposure of the stimulated muscle and that of the contralateral leg to a single conditioning stimulus train (5 Hz) for 25 s via the motor nerve. Isometric tension was measured as were the myosin light chain composition and the activities of the enzymes responsible for phosphorylation and dephosphorylation. A computer simulation of the potential effect of a stimulation-induced disruption of Ca2+ metabolism on phosphorylation was also performed. Chronic stimulation for as little as 1 day eliminated light chain phosphorylation and reduced the myosin light chain kinase activity by approximately 36%. Conversely, phosphatase activity and light chain composition were unaffected. The model demonstrated that a slight depression in the magnitude of the Ca2+ transient could potentially attenuate phosphorylation. The data suggest that phosphorylation of myosin is extremely sensitive to prolonged muscle activity. Furthermore, it appears more likely that this sensitivity is related to regulation of intracellular free Ca2+ than to the other elements of the calmodulin-dependent system for myosin phosphorylation examined.


Attempts to alter the heterogeneous response to ANP in sodium-retaining caval dogs. Chronic caval dogs with ascites were identified as being natriuretic "responders" or "nonresponders" (delta UNaV less than 20 microequiv./min) following an infusion of atrial natriuretic peptide (ANP) (100 ng.kg-1.min-1). To learn more about the factors modulating tubular resistance to ANP, we attempted to convert responders into nonresponders and vice versa by manipulating the physiological environment. To responding dogs, we readministered ANP in the presence of noradrenaline (n = 5), angiotensin (n = 5), indomethacin (n = 4), and adenosine receptor blockage with theophylline (n = 4), and with purposeful reduction of blood pressure (n = 5). To nonresponding dogs, we readministered the ANP in the face of alpha-adrenergic blockade (n = 4), saralasin (n = 4), dipyridamole to block adenosine cellular uptake (n = 5), and elevation of blood pressure (n = 4). In no case were we able to alter the initial natriuretic response to ANP. Binding parameters of ANP receptors in suspensions of renal papillary cells were equivalent in responding caval dogs (n = 6), nonresponding dogs (n = 7), and normal controls (n = 7), as was cGMP generation. We conclude that the tubular resistance to ANP in caval dogs unresponsive to this natriuretic peptide is not due to antagonism from catecholamines or angiotensin but may be due to a post-cGMP problem in signal transduction, or a reduction in the delivery of ANP to the distal nephron.


Neuropathology of the acquired immune deficiency syndrome (AIDS): report of 39 autopsies from Vancouver, British Columbia. Neuropathological findings from 39 acquired immune deficiency syndrome (AIDS) autopsies of primarily neurologically symptomatic patients and 7 brain biopsies from AIDS patients performed at St. Paul's Hospital, Vancouver, British Columbia are reported. Autopsy findings included human immunodeficiency virus-1 (HIV)-type multinucleated giant cell (MNGC)-associated encephalitis seen in 17 patients, toxoplasmosis in 7 patients, and cytomegalovirus encephalitis and/or microglial nodule-associated nuclear inclusions in brain parenchyma in 9 patients. Central nervous system lymphoma was identified in 11 autopsy patients and in 4 of 7 brain biopsies. Infectious processes including HIV encephalitis were seen in 10 of 11 autopsied patients with lymphoproliferative lesions in the brain parenchyma, while 40% of patients without lymphoma had HIV-type MNGC or opportunistic infections. CNS lymphoma was not significantly increased in incidence in patients with a clinical history of zidovudine treatment, but increased duration of survival after the diagnosis of AIDS was associated with increased incidence of lymphoma in both untreated and zidovudine-treated patients. Patients displaying HIV MNGC within microglial nodules had a shorter mean duration of survival after diagnosis of AIDS than those patients with HIV encephalitis with dispersed MNGC, white matter vacuolation, and gliosis.


Malignant astrocytoma of the optic nerve in a child. Malignant gliomas of optic nerve and chiasm are rare, rapidly fatal neoplasms of adulthood. This report documents the occurrence of a malignant astrocytoma of the optic nerve in an 11-year-old boy who 9 years previously had a cerebellar medulloblastoma treated with surgery and irradiation. This malignant optic nerve glioma followed the same aggressive clinical course as that seen in adults, with death 9 months after diagnosis despite surgery and chemotherapy. Radiation may have been an important factor in the development of this malignant tumor which is almost never seen in the pediatric age group.


Trainability of the cardiorespiratory system during childhood. Children have long been suspected of having a diminished capability of responding to endurance training with improvements of maximal oxygen uptake compared to adults. However, studies examining the trainability of children have been beset with methodologic flaws that have precluded firm conclusions about such adult/child differences. Most studies in children that have involved adequate intensity, type, and duration of training have demonstrated the same qualitative changes as would be expected from adult subjects. Some information suggests that children may need a greater exercise intensity than adults to trigger cardiovascular adaptations to training. Other data raise questions regarding differences in autonomic influences on the heart and myocardial function in children that could relate to age-dependent responses to training.


Resistance training during pre- and early puberty: efficacy, trainability, mechanisms, and persistence. Resistance training, under conditions of high intensity and volume loading, is effective in increasing strength in pre- and early pubertal children. The mechanisms underlying strength gain with resistance training in this population have not been established unequivocally. However, resistance training appears to have little if any effect on muscle size (hypertrophy), but it has resulted in neurological (percent motor unit activation and increased integrated EMG activity) changes and changes in intrinsic muscle function (twitch torque), which could account for part of the training-induced increases in voluntary strength. Changes in motor skill coordination (synchronization of muscle action) probably also contribute substantially to resistance-training-induced strength increases in children, particularly for multijoint, complex strength manoeuvres. Most, but not all, studies indicate that pre- and early pubertal children make similar relative strength gains compared to adolescents and adults, but usually demonstrate smaller absolute strength gains following training. Training-induced strength gains appear to decay during detraining, and maintenance training consisting of only one training session per week appears to be ineffective in preserving prior strength gains.


Physical activity among children and youth. The positive influence of physical activity on chronic disease has been well established for the adult population. Much less is known about the relationship between physical inactivity and risk factors for coronary heart disease in children. It has not been clearly established how active children are, nor do we know the factors that determine physical activity patterns among youth. The purpose of this brief review is to describe different techniques used to assess physical activity that are consistent with the operational definition of physical activity. Descriptive information from the National Children and Youth Fitness Studies related to children's level of physical activity is provided which indicates that children are physically active. Nevertheless, it is suggested that the importance of regular physical activity should be emphasized to our youth to promote lifetime activity participation for long-term health and well-being.


Exercise training and body composition in childhood. The influence of exercise training on body composition changes in children is examined in terms of muscle, bone, and fat development. Because of the inexact body composition methodologies in general use and an over-reliance on the two-components system--fat and fat-free body--the extent of changes especially for muscle and bone with exercise training has not been well quantified. With the recent development of new methodologies these limitations may be overcome. The relationship of body composition to health related fitness is also explored with an emphasis on establishing fitness standards for body fatness and the need for research on the etiology of lower back pain and osteoporosis.


A crisis related analysis of perceived spectators' behavior in competition. Spectator behavior in competition is analyzed in terms of spectators' response (positive or negative), or lack of response in reaction to a player's action, or which is independent of a player's action, in a home or away game. This study investigated the perceived contribution of these variables to extreme psychological arousal states (crisis, noncrisis). Forty-eight elite ball-game players responded to a questionnaire asking them to estimate the occurrence probability of the Bayesian combinations among these variables under crisis and noncrisis conditions. An RM-ANOVA procedure revealed that under the crisis condition a negative response, an unsuccessful action, and a home game were rated as more probable, whereas under the noncrisis condition the reverse would be true. The athlete's psychological state was more strongly related to spectator response than to game location or athlete's action. Results are discussed theoretically in reference to the crisis construct.


A validation of a physical activity monitor for young and older adults. The accuracy of the Caltrac accelerometer was studied in 28 young (26.1 +/- 1.1 yrs) and 28 older (64.8 +/- 1.0 yrs; M +/- SEM) men and women to determine its usefulness as an activity monitor for young and older adults. Oxygen uptake was measured by indirect calorimetry while subjects walked on a motorized treadmill at six different speeds while wearing an accelerometer on the hip and on the upper back. The test-retest reliability of the Caltrac was r = .95 and r = .98 for young and older subjects, respectively. In the young group, activity counts correlated highly to net caloric expenditure (absolute minus estimated resting expenditure) (hip vs. kcal/kg: r = .89; back vs. kcal/kg: r = .88) when averaged across speeds, but the relationship was only moderate (back: r = .51; hip: r = .46) when the influence of speed was removed. In the older group the correlations were considerably weaker (back: r = .73; hip: r = .25), and essentially zero when the influence of speed was removed. These data indicate that the Caltrac is a highly reliable accelerometer that is useful for assessing qualitative differences in the level of physical activity among groups, but it lacks accuracy in quantifying energy expenditure in individuals, especially in older adults.


A comparison of predictive tests of aerobic capacity. This study examined the relationship among three predictive tests of aerobic capacity when analyzed within and across gender. Sixty-three active college students (37 males, 26 females) performed the multistage 20-metre shuttle run (SR), the Canadian Aerobic Fitness Test (ST), and the 1.5-mile run (DR). Predicted VO2max (ml.kg-1 x min-1) scores were obtained for each subject. Significant correlations were found among the VO2max results on all possible pairings of tests analyzed as a group, and by gender (p < .001) except for the females' SR and ST pairing (p > .05). One-way analysis of variance was performed for each of the three subject pools with significant F values for the grouped (F = 5.8, p < .01) and male (F = 30.7, p < .01) samples. Tukey HSD post hoc comparison identified significant differences among the SR and ST, and ST and DR VO2 max scores for both samples. However, there were no significant differences between the cell means when females were analyzed separately.


Reliability and validity of three fitness tests for adults with mental handicaps. To examine the reliability and validity of cardiovascular fitness tests, 10 untrained controls and 18 adults with mental handicaps (experimental group) completed five trials on each of four exercise protocols. VO2 max results (M +/- SD) averaged across trials were as follows: Maximal treadmill test, 27.2 +/- 6.2 for the experimental group (E) and 45.5 +/- 8.1 for the control group (C); Maximal shuttle run test, 19.7 +/- 3.4 (E) and 42.1 +/- 6.2 (C); Submax step test, 33.0 +/- 7.6 (E) and 44.5 +/- 7.6 (C); and Submax cycle ergometer test, 36.4 +/- 13.1 (E) and 42.0 +/- 7.1 (C). The four modes had similar VO2 max values across trials with intraclass correlations ranging from r = .90 to .97. To examine validity, the predictive tests were compared with the treadmill VO2 max test. Both the step and shuttle run tests correlated significantly with the treadmill test. The duration of the shuttle run test was unrelated to the exercise intensity. The stepping test is recommended over both the maximum shuttle run test and the submaximum cycling test when measuring the aerobic fitness of adults with mental handicaps.


Effect of time of day on aerobic and anaerobic responses to high-intensity exercise. This study evaluated the effect of time of day on performance of high-intensity, constant-power cycle ergometry by both men and women. Subjects performed all-out cycle ergometer tests in the morning and in the afternoon in randomized order. For all tests, work rate was a constant 5.0 W.kg-1 (women, n = 6) or 6.0 W.kg-1 (men, n = 8). Total work performed was 9.6% greater in the afternoon (mean +/- SE, 348.8 +/- 40.6 J.kg-1) compared to the morning (318.2 +/- 39.5 J.kg-1). The greater amount of work in the afternoon was associated with a 5.1% higher aerobic power and a 5.6% larger anaerobic contribution. There was no interaction between gender and the effect of time of day on the aerobic or anaerobic contributions. These results provide evidence of a circadian rhythm in aerobic and anaerobic responses to high-intensity short-duration exercise, in women as well as in men.


[A qualitative study of the violence in hockey: perceptions of trainers and players]. During semistructured interviews, coaches and players have expressed their perception of violence in hockey through several game situations. The responses reveal that coaches disapprove and even sanction players receiving too many useless penalties, but occasionally congratulate them for a penalized action executed to save a goal. During matches, verbal intimidation is high and not always criticized, especially when it causes the opponent to lose concentration and take a penalty. Body checks have been identified as a main generator of frustration and lack of discipline among players. Data analysis suggests two interventions in training programmes for coaches: the development of teaching material on body checking and on individual counselling techniques to impart sportsmanship attitudes to young players.


Factors affecting university women's basketball coaches' timeout decisions. This study investigated the criteria that coaches of university women's basketball teams used when calling a timeout. Thirty-five of Canada's university coaches of women's basketball responded to Likert scale questions rating the importance of six factors in calling a timeout. Differences due to sex, coaching experience, and team success indicated that some factors were perceived to influence timeout decisions more than others. Specifically, responses from female coaches, coaches with less than 5 years of experience, and coaches whose teams were ranked in the CIAU's top 10 within the last 3 years indicated that they used offensive game events more frequently than their counterparts did. Interactions were also found for Experience x Gender and Experience x Gender x Success in the way that offensive game events were perceived, and for gender and success for the factor attentional state of players.


Revision of the Physical Activity Readiness Questionnaire (PAR-Q). The original Physical Activity Readiness Questionnaire (PAR-Q) offers a safe preliminary screening of candidates for exercise testing and prescription, but it screens out what seems an excessive proportion of apparently healthy older adults. To reduce unnecessary exclusions, an expert committee established by Fitness Canada has now revised the questionnaire wording. The present study compares responses to the original and the revised PAR-Q questionnaire in 399 men and women attending 40 accredited fitness testing centres across Canada. The number of subjects screened out by the revised test decreased significantly (p < .05), from 68 to 48 of the 399 subjects. The change reflects in part the inclusion of individuals who had made an erroneous positive response to the original question regarding high blood pressure. There is no simple gold standard to provide an objective evaluation of the sensitivity and specificity of either questionnaire format, but the revised wording has apparently had the intended effect of reducing positive responses, particularly to the question regarding an elevation of blood pressure.


A case-control study of selected pathogens including verocytotoxigenic Escherichia coli in calf diarrhea on an Ontario veal farm. A case-control study of diarrheal disease in veal calves was conducted over a three month period on a single large veal farm in southern Ontario. One hundred diarrheic calves (cases) were identified by visual examination of their feces. Each case was matched to two nondiarrhetic controls from the same room on the same day, and a fecal sample was obtained from each animal. Fecal consistency of cases and controls was observed daily for one week following sample collection. Control calves which developed diarrhea during that period were excluded from the study. Breed, sex and the date and nature of antimicrobial drugs administered to each calf were recorded. Moisture content of fecal samples was measured by weighing samples before and after oven drying. Samples were screened for verocytotoxigenic Escherichia coli (VTEC) using a Vero cell assay, for enterotoxigenic E. coli (ETEC) using an immunoblot procedure with anti-K99 monoclonal antibodies, and for Salmonella species using modified semi-solid Rappaport-Vassiliadis medium. A latex agglutination test was used to detect rotaviruses, and samples were examined for cryptosporidia using sucrose wet mounts. No VTEC were identified in cases or controls. One calf was positive for Salmonella and three were positive for ETEC. Rotaviruses were detected in four cases and four controls. A significant positive association was found between diarrhea and infection with Cryptosporidium. This study thus provided no evidence of an association between diarrhea and infection with either VTEC, ETEC, Salmonella spp. or rotaviruses in the population examined. On the other hand our results do suggest that Cryptosporidium infection may promote transient diarrheal disease in veal calves in Ontario.


A pathogenesis study of foot-and-mouth disease in cattle, using in situ hybridization. Eight calves were exposed in an aerosol chamber to nebulized foot-and-mouth disease virus. Two control animals were exposed in a similar manner to cell culture media only. Animals were euthanized at intervals and various tissues examined by in situ hybridization using a biotinylated RNA probe corresponding to a portion of the viral gene coding for the polymerase enzyme. By this technique large amounts of viral nucleic acid were found in coronary band, interdigital cleft and tongue as early as six hours postexposure, indicating a very rapid delivery from the portal of entry to the predilection sites for lesion development. This occurred well before the onset of viremia which by virus isolation was not detectable until 30 hours postexposure. The in situ hybridization signal in these tissues decreased in intensity and extent with time to focally positive areas, occasionally surrounding a vesicle. Other epidermal sites not normally thought of as sites for foot-and-mouth lesion development, such as carpus and eyelid, also had some viral nucleic acid detectable at various time intervals. In the lung by in situ hybridization, alveolar septa had viral nucleic acid early in infection (6-18 h postexposure) while later (36-96 h postexposure), the in situ hybridization signal was prominent in alveolar macrophages.


Evaluation of an enzyme-linked immunosorbent assay for the detection of antibodies to caprine arthritis-encephalitis virus in goat serum. An indirect enzyme-linked immunosorbent assay (ELISA), was evaluated for its ability to detect serum antibodies against caprine arthritis-encephalitis virus (CAEV). The ELISA was compared to three other serological immunoassays, agar gel immunodiffusion test (AGIDT), immunoblot assay (IBA), and a fixed-cell immunoperoxidase assay (FCIPA). A total of 511 samples, from 40 farms representing a variety of goat breeds and ages were tested. An estimate of the ELISA sensitivity and specificity was made, relative to combined test results of the three other CAEV serological assays. The degree of agreement of test results among these four assays was evaluated. The number of positives detected by the ELISA, AGIDT, IBA and IPA tests was 193, 154, 204 and 163, respectively. Of the 511 sera tested, 172 were positive to any two or all three of these tests, and were defined as reference positive. A total of 237 samples were negative to all three reference tests, and were defined as reference negative. Relative to these references, the ELISA had a point estimate of 98.3% sensitivity and 97.9% specificity. There was good agreement between the ELISA and the other three assays with a kappa statistic of agreement greater than 0.7 for all three comparisons. The ELISA is therefore considered a suitable assay, with high sensitivity and specificity, for detection of antibodies to CAEV in serum.


Viral infections.

Viral infections. Viral infections may cause serious morbidity as well as death in elderly patients. Of the RNA viruses, influenza virus is the most important pathogen; the majority of influenza-related deaths occur in older patients. Respiratory syncytial virus appears to be gaining increasing importance in elderly persons. Herpes zoster or shingles is caused by the DNA virus, varicella-zoster virus, and its major morbidity in older patients is postherpetic neuralgia.


Clinical stage II non-small cell lung cancer treated with radiation therapy alone. The significance of clinically staged ipsilateral hilar adenopathy (N1 disease). The prognosis of patients with clinically staged hilar nodal involvement (Stage N1) or clinical Stage II non-small cell lung cancer (NSCLC, Stage T1-2N1M0) treated with radiation therapy (RT) alone is not well established. Records of 758 patients with clinical Stage I-III NSCLC treated with RT were reviewed. Sixty-two patients were identified with clinical Stage II NSCLC, and 126 patients had Stage N1 disease. The median survival time (MST) of the 62 patients with clinical Stage II disease was 17.9 months, with 1-year, 2-year, 3-year, and 5-year overall actuarial survival rates of 70%, 33%, 20%, and 12%, respectively. The survival of patients with clinical Stage II disease was significantly better than that of 389 patients with clinical Stage IIIA disease (MST, 11.3 months; P < 0.008) and 267 patients with clinical Stage IIIB disease (MST, 9.8 months; P = 0.0003), but it was similar to that of 40 patients with clinical Stage I lesions (MST, 15.0 months). Patients with performance statuses of 0-1 lived longer than those with a status of 2 or more (MST, 22.8 versus 6.1 months; P < 0.0001). The median survival for patients with N0, N1, N2, and N3 disease was 13.7, 12.6, 10.9, and 9.1 months, respectively. Patients with Stage N0-1 disease (MST, 13.2 months) had significantly improved MST compared with those with Stage N2-3 disease (MST, 10.3 months). The survival of patients with clinical Stage II NSCLC treated with RT alone was significantly better than that of those with clinical Stage IIIA or IIIB disease. It was comparable to that of patients with clinical Stage I lesions. The clinical staging of nodal involvement limited to the ipsilateral hilum does not necessarily portend a worse prognosis than that of patients with clinical Stage N0 disease. The absence of clinically evident Stage N2-3 disease is of significant predictive value for patients with NSCLC treated with RT.


Subclinical ductal carcinoma in situ of the breast. Treatment by local excision and surveillance alone. Mammography has led to earlier detection of subclinical ductal carcinoma in situ (DCIS) of the breast either as nonpalpable calcifications or as an incidental finding in a biopsy performed for another reason. Many women in whom DCIS was detected early may not be destined to have an invasive carcinoma. How should subclinical DCIS be treated if that is the case? What is the role of excision and surveillance only as an alternative to mastectomy or irradiation? All patients with DCIS detected as nonpalpable calcifications or as an incidental finding were eligible for this study. Diagnosis was confirmed, and the histologic subtype was determined. Results of postbiopsy mammography confirmed excision of calcifications; wide local reexcision and assessment of margins was also performed in most patients. The maximum diameter of calcifications considered suitable for this treatment was 25 mm. Between 1978 and 1990, 70 women (72 breasts) were entered into this study (mean follow-up time, 49 months; median follow-up time, 47 months). Of this group, 66% were detected as calcifications and 33% were detected as incidental findings. The recurrence rate was 15.3%. All but one of the patients who experienced a recurrence had the comedo type of DCIS as the initial lesion. Each of the recurrences was of the comedo type. All but one recurrence was at the same site as the primary lesion. None of the patients with DCIS as an incidental finding experienced a recurrence. Excision and surveillance is a reasonable alternative to mastectomy or irradiation for selected women with DCIS that presents as nonpalpable calcifications or as an incidental finding.


Primary small cell carcinoma of the vagina arising in a background of atypical adenosis. Primary small cell carcinoma of the vagina is rare, with only nine cases published to date. The authors report what they believe to be the first case of primary small cell carcinoma of the vagina arising in a setting of atypical adenosis. The tumor was studied by light and electron microscopy and immunohistochemistry. Intracytoplasmic electron-dense neurosecretory type granules were seen, and immunohistochemical demonstration of chromogranin A was documented. These features were found to be similar to small cell carcinomas arising elsewhere in the female genital tract (i.e., cervix, endometrium, ovary, and vulva).


CD30 (Ki-1)-positive, anaplastic large cell lymphoma mimicking gastrointestinal carcinoma. There is great diversity in the clinical presentation of CD30 (Ki-1)-positive, anaplastic large cell lymphoma (ALCL). The authors report two patients in whom the clinical and morphologic features mimicked gastrointestinal carcinoma. Surgical pathology specimens were examined using standard histologic techniques. Paraffin- and frozen-section immunohistochemistry was performed by an avidin-biotin-peroxidase method. The first patient is a 58-year-old man who presented with a constricting mass of the sigmoid colon. Immunohistochemical staining for CD45 (LCA) was negative in formalin-fixed paraffin sections. However, CD45 and CD74 (LN2) were detectable in B5 postfixed material. The second patient is a 44-year-old woman who presented with dysphagia and back pain. Endoscopic examination revealed an ulcerated luminal mass in the proximal esophagus and a gastric ulcer. Mucosal biopsy specimens of the esophagus and stomach contained ALCL, which was CD45 positive. Retroperitoneal lymph nodes contained ALCL infiltrating the sinuses, interfollicular areas, and blood vessel lumina. In addition to CD45 and pan-T-cell antigens, the neoplastic cells expressed CD68 (KP1) and epithelial membrane antigen (EMA). Monoclonal antikeratins were negative, but a polyclonal antikeratin stain was positive. These findings indicate that clinical, morphologic, and some immunologic features of ALCL may mimic gastrointestinal carcinoma.


In vitro measurement of chemosensitivity of human small cell lung and gastric cancer cell lines toward cell cycle phase-nonspecific agents under the clinically equivalent area under the curve. Based on the previous finding that cell killing effects of cell cycle phase-nonspecific agents depend on the concentration-time product (C x T) or the area under the curve (AUC), the authors investigated in vitro cytotoxic effects of nimustine hydrochloride (ACNU) and mitomycin C (MMC) under an experimental condition in which the assay AUC was equivalent to their clinically achievable AUC. The cytotoxic effects of these agents on human cancer cell lines, consisting of 9 small cell lung carcinomas (SCLC) and 10 gastric cancers, were measured by a tetrazolium-based colorimetric assay (MTT assay). These cell lines individually responded to ACNU and MMC in this assay condition. When the authors considered 60% or greater cell kill to be effective, the in vitro response rates of SCLC to ACNU and MMC were 22% (two of nine carcinomas) and 67% (six of nine carcinomas), respectively. The response rates of gastric cancer to ACNU and MMC were 10% (1 of 10 carcinomas) and 40% (4 of 10 carcinomas), respectively. Except for the response of SCLC to ACNU, these in vitro response rates corresponded well to the clinical rates (SCLC to ACNU and MMC, 47% [14 of 30 carcinomas] and 50% [17 of 34 carcinomas], respectively; gastric cancer to ACNU and MMC, 11% [4 of 37 carcinomas] and 30% [63 of 211 carcinomas], respectively). These results suggest that the introduction of the clinically equivalent AUC to the in vitro chemosensitivity test for cell cycle phase-nonspecific agents may improve its clinical predictability.


Urokinase combination chemotherapy in small cell lung cancer. A phase II study. Fifty-one patients with small cell lung cancer (SCLC) were treated with alternating urokinase (UK)-cyclophosphamide-doxorubicin (Adriamycin, Adria Laboratories, Columbus, OH)-vincristine and cisplatin-etoposide-vincristine. UK was given as a loading dose of 3000 micrograms/kg body weight, followed by 3000 micrograms/kg/h for 6 hours. Thoracic irradiation with split technique (46 Gy) and prophylactic cranial irradiation (25 Gy) were administered to responding patients. A second staging was performed in patients exhibiting a clinical complete response (CR) after 1 year. In 27 patients with limited disease, there were 23 CR and 8 partial responses (PR) (CR, 85.1%; 66.2-95.8% at 95% confidence intervals); in 24 patients with extensive disease, there were 17 CR, 4 PR, and 3 cases with progression. Pathologically proven CR were observed in 59.2% patients with limited disease and 33.3% patients with extensive disease. Survival rates were as follows: in patients with limited disease, 1 year, 85.1%; 2 years, 55.5%; and 3 years, 25.9%; in patients with extensive disease, 1 year, 54.1; and 2 years, 16.9%. Median survival times were 26.3 months (patients with limited disease) and 13.3 months (patients with extensive disease). UK-related toxic effects included four episodes of mild to moderate bleeding, one allergic reaction, and one cerebrovascular accident. Myelotoxicity was severe, with a median of two episodes of Grade III-IV (World Health Organization classification) aplasia per patient. These results are consistent with a potential benefit of fibrinolytic therapy in combination with chemotherapy in patients with SCLC with limited disease. Additional trials are indicated.


Immunohistochemistry of Hodgkin and non-Hodgkin lymphomas with emphasis on the diagnostic significance of the BNH9 antibody reactivity with anaplastic large cell (CD30 positive) lymphomas. Morphologic and immunohistologic studies were performed on a series of 213 lymphoma cases, including CD30-positive anaplastic large cell (ALC) lymphomas (45 cases), other CD30-positive (18 cases) or CD30-negative (72 cases) non-Hodgkin lymphomas (NHL), cases of Hodgkin disease (HD) (73 cases), and an additional 5 cases exhibiting features of both CD30-positive ALC and Hodgkin lymphomas. The aim was to assess differential expression by tumor cells of CD30/BerH2, epithelial membrane antigen (EMA), and other monoclonal antibodies, including BNH9, an epithelial and endothelial marker that has been found to be reactive with ALC lymphoma cells. A significantly greater proportion of cases of ALC lymphoma compared with HD exhibited positive results for CD45, EMA, CD45RO, and CD3 in CD30-positive atypical large cells, whereas Reed-Sternberg cells in HD most frequently coexpressed CD15 and CD30 antigens. ALC lymphoma cells reacted with BNH9 MoAb in 10 of the 42 (23.8%) assessable cases, whereas Reed-Sternberg cells reacted with this antibody in 5 of 73 (6.8%) HD cases. Only 3 of 90 (3.3%) NHL cases had positive results for BNH9; they were CD30-negative high-grade or low-grade lymphomas. It was noteworthy that 9 of 10 BNH9-positive ALC lymphomas also were EMA positive. Four of the five cases with morphologic features intermediate between those of HD and ALC lymphoma showed immunohistologic findings (positive results for CD30 and CD15, negative results for EMA, CD45, and BNH9) similar to those frequently observed in the HD cases; conversely, the remaining case showed a profile (positive results for CD30, CD45, and EMA, negative results for CD15) typically observed in ALC lymphomas. This study suggests that the differential expression of CD45, EMA, and CD15 could be used in the distinction of ALC lymphomas and HD, whereas it seems that BNH9 antibody reactivity may be of diagnostic use only in that it reinforces the diagnostic value of EMA expression in the differentiation of these entities.


Verification of isochromosome 12p and identification of other chromosome 12 aberrations in gonadal and extragonadal human germ cell tumors by bicolor double fluorescence in situ hybridization. A diverse group of gonadal and extragonadal human germ cell tumors (GCT) and GCT-derived cell lines was examined for the presence of an i(12p) marker chromosome and/or other abnormalities involving chromosome 12, especially 12p, by bicolor double fluorescence in situ hybridization (FISH). For this purpose three probes, pBS-12, M28, and p alpha 12H8, were used, allowing specific identification of the entire chromosome 12, its short arm, and its pericentromeric region, respectively. The presence of one or more copies of a genuine i(12p) chromosome could be demonstrated in three GCT of the testis, in one ovarian GCT, in one dysgenetic GCT, and in one extragonadal intracranial GCT. Moreover, additional aberrations involving chromosome 12 were shown to be present not only in i(12p) minus but also in i(12p) positive GCT. These data suggest that the occurrence of such aberrations may be a common, although less clearly perceptible and frequent, phenomenon in human GCT.


Effects of genistein on the growth and cell cycle progression of normal human lymphocytes and human leukemic MOLT-4 and HL-60 cells. Genistein (GEN) is an isoflavone known to inhibit both tyrosine protein kinases and DNA topoisomerase II. The effects of GEN on cell proliferation and cell cycle kinetics of human myelogenous leukemia HL-60 and lymphocytic leukemia MOLT-4 cell cultures were studied, and the data were compared to results obtained with normal human lymphocytes stimulated to proliferate with phytohemagglutinin. GEN concentrations greater than 50 micrograms/ml (185 microM) were cytotoxic to HL-60 and MOLT-4 cells following exposure for 24 h; in HL-60 cell cultures, a population of cells with decreased DNA content and nuclear fragmentation characteristic of apoptosis was observed within 8 h. The 50% inhibition concentration after 24 h of exposure for HL-60 and MOLT-4 cells was 8.5 and 13.0 micrograms/ml, respectively. Normal proliferating lymphocytes survived a 24-h exposure of up to 200 micrograms/ml GEN. Short-term (4-8 h) exposures of MOLT-4 or HL-60 cells to 5-20 micrograms/ml GEN resulted in a suppression of cell progression through S or through both S and G2 phases, respectively, while equivalent treatment had no effect on proliferating lymphocytes. A stathmokinetic experiment using MOLT-4 cells revealed that as little as 5 micrograms/ml GEN suppressed cell exit from S to G2 phase by 40%, with a terminal point of action at or near the S-G2 border. Cell progression through the very early portion of G1 phase (G1A, characterized by postmitotic chromatin decondensation) was also suppressed by approximately 40%, whereas cell advancement through the remainder of the G1 phase was not markedly affected. Longer (24 h) exposure of proliferating lymphocytes to 20 micrograms/ml GEN led to an S-phase arrest, while similar treatment of leukemic cells caused cell arrest in G2 phase and an increase in the number of cells entering the cycle at higher DNA ploidy. The mitogen-induced transition of lymphocytes from G0 to G1 phase was extremely sensitive to inhibition by GEN; the 50% inhibition concentration was 1.6 micrograms/ml. The chemotherapeutic value of GEN may be due to the fact that, in terms of cytotoxicity, this agent is more active against proliferating leukemic cells than against normal proliferating lymphocytes. The sensitivity of the G0 to G1 transition in normal lymphocyte cultures and the suppressive effect of GEN on the G1A exit in MOLT-4 cells both suggest that protein kinases involved in chromatin decondensation may be a target of this drug. In light of the observation that lymphocyte stimulation is sensitive to the presence of GEN, the drug is expected to be a strong immunosuppressant.


Chrysotile fiber is a strong mutagen in mammalian cells. Although chrysotile asbestos is a proven human carcinogen, several studies have concluded that these fibers are not mutagenic to cultured mammalian cells. We show here, on the other hand, that when tested using the AL cell system that detects both intragenic and multilocus mutations, chrysotile is indeed mutagenic and comparable in strength to that of gamma-rays. Southern analysis of the induced mutants shows that the majority contains large deletions ranging in size from a few thousand to several million base pairs. Results of our study demonstrate that, while chrysotile may be less durable in vivo than the amphibole fibers such as crocidolites and amosites, it can effectively create genetic damage involved in the cancer process.


p53 gene mutation spectrum in hepatocellular carcinoma. In order to clarify the significance of mutation of the p53 tumor suppressor gene in the genesis and development of human hepatocellular carcinoma (HCC) in an aflatoxin B1 low-exposure area, the spectrum, i.e., incidence, type, and site, of p53 gene mutations was examined in 169 tissue samples resected mainly from Japanese patients using single-strand conformation polymorphism analysis and direct sequencing. Forty-nine tumors (29%) showed a p53 mutation (39 point mutations and 10 frameshifts). The point mutations comprised 18 transitions, only 4 of which occurred at CpG sites, and 21 transversions. Two evolutionarily conserved domains, IV and V, contained 65% of all mutations and codon 249 was the most frequent mutation site (7/49). The spectrum of p53 mutation did not differ among HCCs in relation to the type of hepatitis virus infection, sex, age, and background liver disease of patients, tumor size, or presence of metastasis, but incidence and site were significantly associated with the degree of differentiation of cancer cells. In poorly differentiated HCC, p53 mutation was frequent (54%) and clustered on domains IV and V, whereas in well or moderately differentiated HCC, the mutation was less frequent (21%) and equally distributed on domains II to V. Restriction fragment length polymorphism analysis revealed loss of heterozygosity on chromosome 17p in 55 (69%) of 80 informative cases and in 34 (95%) of 36 cases with p53 mutation. Therefore, p53 gene mutation is suggested to occur independently of the type of viral infection or status of preexisting liver disease and to occur preferentially in moderately and poorly differentiated HCCs in association with or after loss of another p53 allele as a late event of HCC progression.


Accessibility to DNA in carcinoma chromatin is promoted by nanomolar okadaic acid: effect on AT-rich DNA binding proteins. Differential accessibility to DNA in tumor cell chromatin is important to growth, differentiation apoptosis, and the targeting of DNA modifying drugs. We now show that endonuclease accessibility to DNA in the nuclei of A431 human carcinoma cells is increased within 90 min by nontoxic nanomolar levels of okadaic acid, known to inhibit protein phosphatase 2A. This genomic hypersensitivity was partly enhanced by joint treatment with epidermal growth factor and okadaic acid but did not appear without the latter. Nuclei with greater DNA susceptibility showed a decrease in M(r) 80,000 DNA binding protein doublet specific for dAT-rich sequences concurrent with the "apparent" hyperphosphorylation of a M(r) 70,000 nuclear matrix protein. We propose that some of the tumor-promoting effects of okadaic acid may be partly associated with its ability to promote genomic susceptibility.


Molecular cloning of rat Wilms' tumor complementary DNA and a study of messenger RNA expression in the urogenital system and the brain. In an effort to study the molecular basis of kidney development and carcinogenesis, we isolated complementary DNA clones of the rat homologue of the human Wilms' tumor gene, WT-1. When compared to the predicted sequence of the human WT-1 polypeptide, the rat WT-1 amino acid sequence is highly conserved (> 97%), except for the loss of one amino acid. In situ mRNA hybridization experiments localized WT-1 expression to the glomerular cells in the kidney during embryogenesis and the Sertoli cells of the testis. Similar to expression in humans and mice, WT-1 mRNA expression in the rat kidney and testis is developmentally regulated. In addition, two novel sites of specific high level WT-1 mRNA expression were seen. One is an area in the spinal cord where high level expression occurs throughout embryonic development. The second is the area postrema in the brain where localized expression continues through adulthood, suggesting a functional role for WT-1 in rat brain.


Tumor-associated M(r) 34,000 and M(r) 32,000 membrane glycoproteins that are serine phosphorylated specifically in bovine leukemia virus-induced lymphosarcoma cells. Tumor-associated antigens that are expressed in tumor cells of cattle with enzootic bovine leukosis (EBL) were analyzed previously by use of 13 monoclonal antibodies. We biochemically identified one of the tumor-associated antigens, which is recognized by the c143 monoclonal antibody, as two glycoproteins, each having an apparent molecular weight of 32,000 or 34,000. These glycoproteins were found in the plasma membrane of peripheral blood lymphocytes of both bovine leukemia virus (BLV)-free normal and BLV-infected cattle. With the progression of EBL, the proportion and the absolute number of cells positive for the tumor-associated antigen increased. Moreover, the level of the M(r) 34,000 component, which was susceptible to cell-surface labeling, increased over the level of the M(r) 32,000 component. Partial proteolytic peptide mapping with V8 protease and deglycosylation analysis revealed that the two glycoproteins most likely have an identical M(r) 30,000 polypeptide portion but have different N-linked oligosaccharide portions. Both glycoproteins were found to be phosphorylated at serine residue(s) in EBL-derived B-lymphoid cell lines and in tumor cells and peripheral blood lymphocytes from cattle with EBL, but not in peripheral blood lymphocytes from BLV-free normal cattle and BLV-infected cattle without any evidence of tumor, suggesting that the phosphorylation of these glycoproteins is related to the transformed state of the BLV-infected B-lymphoid cells.


Pregnancy-dependent to ovarian-independent progression in mammary tumors delineated in primary culture: changes in signal transduction, growth factor regulation, and matrix interaction. A model for the evolution of hormone-independent tumors from a pregnancy-dependent precursor is exemplified by the TPDMT-4[pregnancy-dependent (PD)] and T4OI96 [ovarian-independent (OI)] in vivo tumor lines developed in DDD mice. In vivo, the OI tumor grows rapidly in virgin mice and is more anaplastic than the PD tumor which, in virgin mice, grows as a hyperplastic alveolar gland from which tumors arise only during pregnancy. The regulation of the proliferation of these two tumors was compared in primary culture using a three-dimensional, serum-free, collagen gel cell culture system. In medium containing insulin, the growth of cell organoids from PD tumors was stimulated by the same factors that stimulate the growth of normal mammary epithelial cells from virgin or pregnant mice. These factors include progesterone and prolactin (but not estrogen), epidermal growth factor, basic fibroblast growth factor, linoleic acid, cyclic AMP, prostaglandin E2, phosphatidic acid, and lithium. Most of these tumors (about 80%) could not grow in medium with only insulin present; of those that did, growth was slow and was stimulated further by the above agents. PD tumor cells formed stellate colonies in the collagen matrix similar to those of normal cells. These findings show that the growth regulation of PD tumor cells is, in all aspects examined, similar to that of normal cells. In addition, the capacity for growth in the presence of only insulin (more autonomous growth) is not necessarily accompanied by deletions in responses to growth factors or hormones, or in lipid response pathways. In contrast, organoids from OI tumors needed only insulin for growth. The growth of some of these tumors was stimulated further by only basic fibroblast growth factor and phosphatidic acid. Cyclic AMP and agents that elevate intracellular cyclic AMP inhibited growth, the opposite of the response seen in normal or PD cells. OI organoids grew as cell masses rather than as stellate structures. These data show that while PD tumors are remarkably similar to normal cells in the regulation of their proliferation, OI tumors have incurred multiple defects in growth-regulatory pathways possibly including the production of autocrine growth factor(s). In addition, the ability of OI tumor cells to adhere to the collagen gel matrix and undergo normal morphogenesis is reduced. These results may highlight specific events required for the progression from ovarian dependency to ovarian independency related to the hormonal regulation of growth.


Mapping chromosomal breakpoints of Burkitt's t(8;14) translocations far upstream of c-myc. To analyze the region upstream of c-myc, a number of novel probes were established. These were generated by chromosomal walking starting from the breakpoint of the chromosomal translocation of the B-cell line 380 and by cloning the breakpoint of the translocation of the Burkitt lymphoma cell line IARC/BL72. Using the newly isolated probes a detailed physical map of 500 kilobases of the region upstream of c-myc was established applying pulsed-field gel electrophoresis. The chromosomal breakpoint of IARC/BL72 cells was mapped to a site 55 kilobases 5' of c-myc. A region 20 kilobases in length and containing the breakpoints of 380, EW36, P3HR-1, and Daudi cells was identified 170-190 kilobases upstream of c-myc. In addition the HPV18 integration site in HeLa cells was located between 340 and 500 kilobases 5' of c-myc. The probes were used to define the c-myc amplification units in Colo320-HSR and HL60 cells as well as in four cases of small cell lung cancer. Evidence is provided that the amplicon of HL60 cells is discontinuously organized.


T-cell receptor beta chain gene rearrangement in acute myeloid leukemia always occurs at the allele that contains the undermethylated J beta 1 region. The epigenetic phenomenon could play a role in the interaction between chromatin and DNA-binding enzymes, allowing us to consider an association between the phenomenon and gene rearrangement. The correlation between methylation status and rearrangement of the T-cell receptor (TCR) beta chain gene in leukemia cells obtained from patients with acute myeloid leukemia (AML) was examined. All of the AML patients with a TCR-beta rearrangement had hypomethylated CCGG sequences within the J beta 1 region on the rearranged allele, while the germline allele had completely methylated CCmeGG sequence in this region, indicating a strong association between hypomethylation status and rearrangement of the TCR beta chain gene. In the DNA from AML patients with or without a TCR-beta rearrangement, the C beta 2 region contained completely methylated CCmeGG sequences, even though they express T-cell-associated antigens, including CD7; this pattern is quite different from that observed in T-cell neoplasias. Moreover, some AML patients showed a TCR-beta rearrangement without the presence of immunoglobulin heavy-chain gene rearrangement, suggesting that TCR beta chain gene involvement in AML is required for unknown factors other than common recombinase activity.


Sensitivity of resistant human tumor cell lines to tumor necrosis factor and adriamycin used in combination: correlation between down-regulation of tumor necrosis factor-messenger RNA induction and overcoming resistance. A number of human tumor cell lines of various histological origin were examined for their sensitivity and resistance to tumor necrosis factor-alpha (TNF) and Adriamycin (ADR). Six ovarian lines, and one each of a renal, lung, and B-cell line, were tested for putative mechanisms of resistance to these agents. Cytotoxicity resulting from TNF or ADR showed no overall correlation in these lines. The combination of TNF and ADR produced enhanced cytotoxicity against these tumor lines and furthermore resulted in overcoming the resistance of TNF or ADR alone or in combination. A proposed mechanism of TNF resistance in tumor cells is the endogenous production of TNF mRNA and protein. There was a positive correlation between resistance to TNF and the constitutive production of TNF mRNA and protein. The TNF-resistant lines that did not constitutively produce TNF mRNA and protein and the three TNF-sensitive tumor lines exhibited up-regulation of their TNF mRNA in the presence of TNF or phorbol myristate acetate/ionophore, but did not secrete any detectable protein. Due to the enhanced cytotoxicity seen with the combination of TNF and ADR, the effect of this combination on the level of TNF mRNA was examined. ADR alone reduced the constitutive level of TNF mRNA and in combination with TNF reduced the level of induction produced by TNF. This down-regulation of TNF mRNA by ADR may play a role in the enhanced cytotoxicity seen with the combination of these 2 agents.


Cellular localization of the folate receptor: potential role in drug toxicity and folate homeostasis. In the past year, gp38, a glycosyl-phosphatidylinositol linked membrane protein that is overexpressed in some malignant tissues, has been shown to be the folate receptor. Using immunohistochemical techniques with the monoclonal antibody MOv19 against gp38, we evaluated the cellular localization of folate receptors in normal human tissues, which are potential target sites for drugs that utilize this uptake mechanism. The choroid plexus was intensely positive with staining limited to the epithelium, which in some foci had a distinct bilaminar pattern limited to the luminal and basal surfaces. The epithelium of the fallopian tube, uterus, and epididymis was highly immunoreactive. The acinar cells of the breast, submandibular salivary, and bronchial glands also showed intense staining as did the trophoblastic cells of the placenta. In the kidney reactivity was localized to the proximal tubules. Lung alveolar lining including type I and II pneumocytes stained intensely. Limited but focal reactivity was noted in the vas deferens, ovary, thyroid, and pancreas. This study in conjunction with previous work showing marked overexpression of folate receptor in some malignant cells suggests that the folate receptor may be an important target for diagnostic or therapeutic exploitation and indicates sites of potential drug toxicity.


T cell recognition of Epstein-Barr virus associated lymphomas. Epstein-Barr virus, a lymphotropic herpesvirus of humans, has potent B cell growth transforming activity yet persists in the lymphoid tissues of most individuals as a lifelong asymptomatic infection. Virus induced B cell growth transformation in vitro is associated with the expression of a limited set of viral genes encoding six nuclear antigens (EBNA 1, 2, 3A, 3B, 3C and LP) and two latent membrane proteins (LMP 1, 2). Healthy virus carriers possess strong EBV specific CTL memory that can be reactivated in vitro. Here, we summarize experiments in which the antigenic specificities of these HLA class I restricted memory CTL responses have been mapped in a range of individuals with different HLA backgrounds. Of the known EBV latent proteins, EBNA 3A, 3B and 3C are frequently the dominant targets for such responses, but examples of responses directed against epitopes of EBNA 2, EBNA-LP or the LMP have been identified; by contrast, CTL responses against epitopes of EBNA 1 have not been observed. Epstein-Barr virus is associated with at least three malignancies of lymphoid origin--immunoblastic lymphomas of the immunosuppressed, endemic Burkitt's lymphoma and a subset of Hodgkin's disease. The immunoblastic lymphomas express the complete spectrum of EBV coded latent proteins and a cellular phenotype similar to that of in vitro transformed B lymphoblastoid cell lines; accordingly, they remain sensitive to EBV specific CTL recognition. Endemic BL cells are not recognized by such CTL, and at least three consistent features of this tumour could contribute to immune escape: (a) allele specific downregulation of HLA class I antigen expression, (b) absence/low expression of cellular adhesion molecules and (c) restriction of EBV latent protein expression to EBNA 1 only. The relative importance of these three features of the BL cell phenotype with regard to sensitivity to CTL recognition is re-interpreted in the light of recent results. Finally, the pattern of virus latent protein expression in EBV positive Hodgkin's disease is described, and the possibility of EBV specific CTL control against this tumour is discussed.


Progression from steroid sensitive to insensitive state in breast tumours. Progression from a hormone sensitive to insensitive tumour can take place by several independent pathways, some of which involve loss of receptor, whereas, others may be due to the presence of abnormal receptors. Examples now exist of insensitivity despite seemingly normal receptors, such cells being generated either by spontaneous changes or by steroid deprivation. Breast cancer is a multifactorial set of diseases with steroids being only one component of the jigsaw; it is likely that further mechanisms of response loss will be identified when experimental systems are refined and additional parts of the jigsaw identified.


Retinoic acid receptors in normal growth and development. Retinoic acid receptors are ligand activated transcription factors that have a variety of important roles in normal development, through regulating the transcription of other developmental control genes. The great diversity of RA mediated developmental events is made possible through the existence of three receptor subtypes, each of which has a number of isoforms. The mechanism of gene regulation is thought to involve binding to an RA response element on the target gene, although in practice, few genes known to be activated by RA have so far been demonstrated to include such an element in the promoter region. Differential spatiotemporal expression patterns of the various subtypes have been described in mouse embryos, including very fine differences within different subpopulations of cells of the same tissue. In addition to qualitative differences in the gene activation responses to RA through spatiotemporal differences in RAR expression patterns, local differences in RAR activation may be modulated by cytoplasmic binding proteins for retinol and RA, through quantitative control of ligand availability. The developmental control genes known to be activated by RA include the 3' members of each of the homoeogene clusters, and it is likely that others will be identified in the near future.


Retinoic acid receptor alpha in acute promyelocytic leukaemia. Acute promyelocytic leukaemia has two highly specific particularities: a t(15;17) chromosomal translocation and the ability of a differentiation inducer all-trans-RA, to revert the malignant phenotype both in vitro and in vivo. Molecular characterization of the t(15;17) translocation has shown that it fuses a previously unknown zinc finger encoding gene, PML, to the RAR alpha, suggesting a link between the molecular mechanism of transformation and of RA dependent differentiation. The PML/RAR alpha fusion receptor--which is functionally altered--may block RA target genes, impair RA mediated differentiation and lead to transformation. Alternatively, or in addition, the PML transduction pathway may also be affected. Although it is clear that RA treatment must relieve APL cells from differentiation arrest, so far no model can satisfactorily account for this effect.


Mechanistic interrelationships between two superfamilies: the steroid/retinoid receptors and transforming growth factor-beta. The fields of endocrinology and peptide growth factors can no longer be studied independently, nor can hypotheses about the modes of action of hormones and growth factors be constrained by outmoded classic definitions. Although much is still to be learned about the mechanisms of induction of TGF-beta activity by steroids/retinoids, about the involvement of specific nuclear receptors and about the factors regulating the differential effects of steroids/retinoids on the various isoforms of TGF-beta, it is clear that the identification of the interrelationships between these two superfamilies of multi-functional effectors has heralded a new era of discovery of the underlying regulatory events controlling cellular proliferation and differentiation.


Peroxisome proliferators: a model for receptor mediated carcinogenesis. Peroxisome proliferators are a diverse group of rodent hepatocarcinogens, which include hypolipidaemic drugs, plasticizers and herbicides. A member of the nuclear hormone receptor superfamily has been identified that can be activated by peroxisome proliferators. The receptor is therefore termed the peroxisome proliferator activated receptor (PPAR). Three Xenopus nuclear hormone receptors have been described that are closely related to the mouse PPAR and can all be activated by peroxisome proliferators. Therefore, a family of PPARs that is possibly important in mediating the action of some hypolipidaemic drugs may exist in many species including mice and humans. The most widely used marker of peroxisome proliferator action is the peroxisomal beta-oxidation enzyme acyl CoA oxidase. PPAR recognizes a specific peroxisome proliferator response element located in the acyl CoA oxidase gene promoter. It is therefore argued that the hypolipidaemic and carcinogenic effects of peroxisome proliferators may be mediated by PPAR in a manner similar to that of steroid hormone action.


GABA-active steroids: endogenous modulators of GABA-gated chloride ion conductance. Naturally occurring 3-alpha-hydroxy ring A-reduced metabolites of progesterone and deoxycorticosterone and their synthetic analogues bind to specific sites within the hydrophobic channel domain of the GABAA receptor complex. Acting at these sites, these ligands function as positive allosteric effectors of the complex; they potentiate GABA-stimulated membrane chloride ion conductance, enhance the binding of [3H]muscimol and [3H]flunitrazepam, and displace the binding of [35S]t-butylbicyclophosphorothionate ([35S]TBPS), a channel ligand that is a specific marker of the GABA-associated chloride ionophore. Moreover, steroid metabolites (namely pregnenolone sulfate and dehydroepiandrosterone sulfate) have been identified that display properties of GABA-negative allosteric effectors. The identification of this membrane-associated steroid binding should stimulate development of new classes of anxiolytic, sedative-hypnotic, anticonvulsant, anesthetic, and muscle-relaxant medications that may be devoid of many of the side effects associated with benzodiazepines and barbiturates. Also, elucidation of the physiologic role of this binding site should contribute both to our understanding of endogenous mechanisms for modulating inhibitory neurotransmission, and the pathophysiologic role of the GABAA receptor complex in a variety of neuropsychiatric disorders.


Clinical and pharmacokinetic effects of a diet rich in insoluble fiber on Parkinson disease. In this study, the effects of a diet rich in insoluble fiber (DRIF) on motor disability and the peripheral pharmacokinetics of orally administered L-dopa in Parkinsonian patients with marked constipation are analyzed. We found a useful effect of a DRIF on plasma L-dopa concentration and motor function. The greatest effect on the plasma L-dopa levels was found early (at 30 and 60 min) after oral administration. There was a relationship between the improvement of constipation and the higher bioavailability of L-dopa. DRIF can be a coadjuvant treatment in patients with Parkinson's disease.


Interaction of zinc with dental mineral. As some currently available toothpastes contain zinc compounds, the reaction of zinc with dental mineral and its effect on crystal growth rates were studied using three synthetic calcium-deficient hydroxyapatites (HAP) as being representative of dental mineral. Zinc was readily acquired by all HAP samples in the absence of added calcium, the amount adsorbed being proportional to the HAP surface area; about 9 mumol Zn/m2 was adsorbed at high zinc concentrations. As zinc was acquired, calcium was released, consistent with 1:1 Ca:Zn exchange. Soluble calcium reduced zinc uptake and similarly, calcium post-treatment released zinc. Pretreatment of HAP with 0.5 mM zinc reduced its subsequent ability to undergo seeded crystal growth, as did extracts of a toothpaste containing 0.5% zinc citrate, even in the presence of saliva. The reverse reaction, i.e. displacement of adsorbed zinc by salivary levels of calcium, however, indicates the mechanism by which zinc can reduce calculus formation in vivo by inhibiting plaque mineralisation without adversely affecting the anti-caries effects of fluoride.


Analysis of binding and activating functions of the chick muscle acetylcholine receptor gamma-subunit upstream sequence. 1. The skeletal muscle acetylcholine receptor comprises several subunits whose coordinated expression during myogenesis is probably controlled by cis elements in the individual subunit genes. We have previously analyzed promoter regions of the alpha and delta genes (Wang et al., 1988, 1990); to gain further insight into receptor regulation, we have now studied the promoter of the chick muscle gamma-subunit gene. 2. This analysis was faciliated by the close upstream proximity of the coding region of the delta-subunit gene and the consequent brevity (740 bp) of the untranslated linker connecting the two genes (Nef et al., 1984). Nuclease protection and primer extension analysis revealed that transcription of the gamma-subunit gene starts at position 56 upstream of the translational initiation site. 3. Nested deletions of the promoter region were employed to identify functionally important elements. A 360-bp sequence (-324 to +36) was found to activate transcription, in a position- and orientation-independent manner, during myotube formation. This sequence comprises 5 M-CAT (Nikovits et al., 1986) similarities and contains, at positions -52/-47 and -33/-28, two CANNTG (Lassar et al., 1989) motifs. 4. Binding experiments were performed by means of gel retardation, gel shift competition, and footprint analysis. The CANNTG motifs were found to bind MyoD and myogenin fusion proteins and to interact with proteins in nuclear extracts from cultured myotubes. 5. Point mutations in the CANNTG motifs revealed that these elements are crucial for full promoter activity in myotubes and essential in fibroblasts cotransfected with a myogenin expression vector. 6. We conclude that the activity of the gamma-subunit gene is determined largely by E boxes, which in vivo are likely to be activated by MyoD family proteins; in addition, other transactivators such as the M-CAT binding protein presumably play a role. Both CANNTG elements and M-CAT motifs are also present in the alpha- and delta-subunit enhancer and may therefore account for the coordinate expression of the three subunits during muscle differentiation.


Accumulation of different c-erbA transcripts during rat brain development and in cortical neurons cultured in a synthetic medium. 1. Accumulation of different c-erbA transcripts was studied, during rat brain maturation and in cortical neurons differentiating in a serum-free medium, by quantitative Northern blot hybridization. 2. The alpha and beta forms of c-erbA mRNAs exhibit different patterns of accumulation, with a precocious increase in the alpha forms compared with the beta forms both in vivo and in culture. 3. erbA alpha 2 mRNA (2.6 kb) is by far the predominant form, with a maximum at birth (PO). 4. The accumulation patterns of both alpha and beta forms show discrete differences in isolated neurons compared to brain cortices; in particular the pattern of alpha 2 mRNA accumulation in culture suggests its predominant localization to neurons. 5. The presence of T3 in the culture medium does not have significant effects on the level of any of erbA mRNAs. 6. Possible implications and relationships with neuronal terminal differentiation are discussed.


Effects of imidazole compounds on catecholamine release in adrenal chromaffin cells. 1. Effects of imidazole compounds and guanabenz on the stimulus-evoked release of catecholamine (CA) were studied in cultured bovine adrenal chromaffin cells. 2. Clonidine, oxymetazoline, phentolamine, chlorpheniramine, and guanabenz inhibited acetylcholine (ACh)-evoked CA release in a dose-dependent manner, but not high K(+)-evoked release. 3. The inhibition by these compounds was not antagonized by nonimidazole and nonguanidine alpha 2-antagonists (yohimbine and phenoxybenzamine) but was significantly antagonized by tolazoline (imidazole alpha 2-antagonist) and cimetidine (imidazole H2-antagonist). Moreover, tolazoline by itself augmented the ACh-evoked, but not the high K(+)-evoked, CA release. 4. Although chlorpheniramine and cimetidine are antagonists for H1 and H2 histaminergic receptors, the site of action for these compounds in our results seemed to differ from the histamine receptors. 5. These results suggest that the inhibitory action of imidazole compounds and guanabenz on ACh-evoked CA release in adrenal chromaffin cells is mediated through an imidazole receptor. Adrenal chromaffin cells may contain an endogenous clonidine-displacing substance (CDS) which has been found in adrenal gland and brain as an endogenous ligand for imidazole receptors. Thus, CDS may have a regulatory role in the stimulus-secretion coupling in these cells.


[IgA antibodies against Epstein-Barr virus capsid antigen]. The authors describe infection with the Epstein-Barr virus (EBV) which is associated with the formation of various types of antibodies--heterophil (HP) antibodies as well as antibodies against individual EBV antigens. They evaluate the contribution of a recently elaborated and introduced method used for detection of antibodies against the capsid antigen of the EBV class IgA (anti-VCA IgA) by indirect immunofluorescence to the diagnosis of acute and chronic EBV infections. Anti-VCA IgA are useful in the diagnosis of EBV infections if the patient has no longer HP and anti-VCA IgM, but only anti-VCA IgG. If, however, in addition to anti-VCA IgA also anti-VCA IgA are present, their presence can be the only detectable sign of a recent EBV infection and also a sign of reactivation of chronic EBV infection.


Regeneration of autotransplanted splenic tissue at different implantation sites. Inbred animals (Lewis rats) were used to investigate the regeneration of autologously implanted splenic tissue at intra-omental and subcutaneous sites. Quantitative immunohistology with monoclonal antibodies against lymphocytes and macrophages was performed to analyse the cell density of red pulp (RP), periarteriolar lymphoid sheath (PALS), marginal zone (MZ) and follicle, 7-180 days after transplantation. Antigenic, allogeneic and mitogenic stimulation and Northern blotting were also performed. Transplant groups differed from spleen only in the reduced size of PALS; however, quantitative analysis demonstrated subtle differences between spleen and transplants. The cell density of B-cells and ED-1+ macrophages was reduced in the RP, Tsupp/cyt-cells were decreased and B-cells increased in PALS, and B-cells and Thelper-cells reduced in the MZ. No differences could be detected between the transplant groups. Flow-cytometric analysis of cell suspensions from spleen and transplants revealed a reduction of T-cells (OX-19+), MHC-I and transferrin-receptor-bearing cells in both transplant groups, and a decrease in the number of Thelper-cells and ED-3+ macrophages in subcutaneous transplants. Both transplant groups were defective regarding the allogeneic and pokeweed mitogen response. Aberration of the lipopolysaccharide response was restricted to subcutaneous transplants, which additionally showed abnormal expression of interferon-gamma, interleukin-5 and interleukin-6 mRNA. Thus, subtle alterations of the newly developed microenvironment and/or lymphocyte-homing may influence the regeneration of splenic tissue; the implantation site may represent an important parameter in functional reorganisation.


Immunocytochemistry of somatotrophs, gonadotrophs, prolactin and adrenocorticotropin cells in larval sea bream (Sparus auratus) pituitaries. The chronological appearance of endocrine cells in the pituitary of sea-bream (Sparus auratus) larvae was studied using antisera against salmon prolactin, trout growth hormone, salmon gonadotropin and N-terminal human adrenocorticotropin. The larval pituitary (1-12 days after hatching) was oval in shape and was composed of a dense mass of cells with few neurohypophysial fibres. By 60 days after hatching it began to resemble the adult and was divisible into a distinct rostral pars distalis containing prolactin and adrenocorticotropin cells; a proximal pars distalis containing somatotrophs and gonadotrophs and a pars intermedia. Cells immunoreactive with antisera against growth hormone were observed immediately after hatching (2 days post-fertilization). Weakly staining prolactin cells were observed 2 days later in the region corresponding to the rostral pars distalis. Cells immunoreactive with antigonadotropin and anti-adrenocorticotropin sera were observed in the pituitary 6 and 8 days after hatching, respectively. All the cell-types studied were immunoreactive from the time they were first identified until the final samples 90 days after hatching.


Ecto-ATPase/phosphatase activity in the olfactory sensilla of the spiny lobster, Panulirus argus: localization and characterization. Electrophysiological studies have shown that the olfactory organ (antennule) of the spiny lobster, Panulirus argus, has chemoreceptors that are selectively excited by adenine nucleotides in seawater. Biochemical studies have revealed that these same nucleotides can be rapidly dephosphorylated by ectoenzymes associated with the olfactory sensilla (aesthetascs). In this study the distribution of ecto-ATPase/phosphatase activity within aesthetascs was determined cytochemically and the nature of the adenine-nucleotide dephosphorylating activity was dissected biochemically. Cytochemically, the distribution of ATP-dephosphorylating activity was similar to that shown previously for AMP and beta-glycerol phosphate; i.e., cerium phosphate reaction product was specifically localized to the transitional zone where the sensory dendrites develop cilia and branch to form the outer dendritic segments. Unlike the dephosphorylation of AMP and beta-glycerol phosphate, Mg2+ or Ca2+ was required for ecto-ATPase/phosphatase activity. Biochemical measures of both AMP- and ATP-dephosphorylating activity within aesthetascs corroborated the cytochemical evidence that these activities are localized to the transitional zone. A major portion of the AMP dephosphorylation (about 67%) derives from nonspecific alkaline phosphatase activity that is insensitive to levamisole and L-bromotetramisole. In contrast, nonspecific phosphatase activity accounted for a much smaller part of the ATP dephosphorylation (about 15%). Ectoenzymatic activity in the transitional zone may be an important means of removing excitatory/inhibitory nucleotides from this region.