Giant cell arteritis-a systemic spectrum including temporal arteritis and polymyalgia rheumatica. Polymyalgia rheumatica and giant cell arteritis are closely associated disorders that may pose serious threats to the elderly. While the etiological factors remain unknown, these clinical entities may represent different expressions of a common disorder. Recent evidence suggests that autoimmunological involvement of the internal elastic lamina of the large and medium-sized arteries might play an importent role in pathogenesis. Genetic and environmental factors may also contribute. Polymyalgia rheumatica and giant cell arteritis have subtle and protean manifestations and formes frustes occur, contributing to delays in diagnosis unless the clinician holds a high index of suspicion while treating the elderly. The sequelae, including generalized disability, blindness, myocardial and cerebrovascular catastrophies can probably be avoided by timely and prudent use of corticosteroids. These are not uncommon disorders are among the most amenable to treatment of the rheumatological diseases. Family physicians must be diligent in their efforts to diagnose these disorders since early generalized and focal signs and symptoms frequently precede the catastrophic events.


Complete right bundle branch block and acute myocardial infarction an ominous sign. Complete right bundle branch block associated with a normal QRS axis and a normal PR interval was observed in 2.8 percent of 500 consecutive acute myocardial infarction patients admitted to the coronary care unit of a community teaching hospital. This experience and a review of the literature reveals that this unifascicular block complicating acute myocardial infarction is typically associated with (1) an elderly patient population, (2) a hospital mortality rate in excess of 50 percent, and (3) a "new" onset complete right bundle branch block which carries a higher mortality than when this electrocardiographic pattern preexists the acute infarction.


Effect of topical corticosteroids on symptoms of clinical sunburn. Topical corticosteroids have been advocated in the treatment of sunburn. This therapy was evaluated by means of a random, double-blind study in which subjects who had acquired acute natural sunburn served as their own controls. Subjects applied fluocinolone cream to an acutely exposed area of their body and the inert carrier base to a symmetrical, similarly exposed area twice a day for five days while avoiding further exposure. The subjects recorded by means of a rating scale the progression, at both sites, of redness, pain, blistering, swelling, and peeling. The natural course of mild to moderate sunburn to symptomatic resolution was observed to be about three to five days. There was no significant difference in response between the steroid-treated area and the inert-base-treated area.


A systematic approach to brief psychological intervention in the primary care setting. A systematic approach to psychological intervention that can be used by the primary care physician is presented. The approach specifies the sequence the physician takes when providing therapy: (1) identify the context of the patient's complaint(s), (2) assess the patient's current status, (3) develop a therapeutic plan, and (4) implement a therapy approach. These steps are based on four current status categories-functional, ambiguous, crisis, and dysfunctional-that describe the patient's life situation. Each category serves as a gateway for planning and for carrying out therapy. Four types of psychological intervention-short-term therapy, structured assessment, crisis intervention, and referral-are described and methods for implementing each activity are given. The advantages of the schema and a brief critique are presented.


The family as patient: preliminary experience with a recorded assessment schema. Although family medicine has become a well-established discipline, emphasis more often than not continues to be on the individual and not the family. It is the intent of this paper to expand the focus of physician concern beyond the presenting symptom and, indeed, beyond the individual patient, to include the concept of family as patient. The family is discussed as part of a complex interactional system of stress and support factors both from within the family itself and from the wider social system of which it forms a part. A three-dimensional model for the comprehensive psychosocial evaluation of family functioning in health and illness has been presented both diagrammatically and descriptively. Most importantly, a simple set of questions is provided as an instrument to assist the physician in the assessment of family functioning in health and illness.


The no-show patient in the model family practice unit. Appointment breaking by patients causes problems for the physician's office. Patients who neither keep nor cancel their appointments are often referred to as "no shows." Twenty variables were identified as potential predictors of no-show behavior. These predictors were applied to 291 Family Practice Center patients during a one-month study in April 1977. A discriminant function and multiple regression procedure were utilized ascertain the predictability of the selected variables. Predictive accuracy of the variables was 67.4 percent compared to the presently utilized constant predictor technique, which is 73 percent accurate. Modification of appointment schedules based upon utilization of the variables studies as predictors of show/no-show behavior does not appear to be an effective strategy in the Family Practice Center of the Community Hospital of Sonoma County, Santa Rosa, due to the high proportion of patients who do, in fact, show. In clinics with lower show rates, the technique may prove to be an effective strategy.


Analysis of after-hours calls and visits in a family practice. The after-hours encounter is an important aspect of primary care. This study examined the pattern of patient behavior after hours in a rural Ontario setting serving a mixed white and native population. Of the 83 telephone calls and 59 visits over a six-week period, approximately 60 percent were from females (52 percent of the clinic population), and 70 percnet were for new problems, with acute infection accounting for nearly one third of the contacts. Forty-seven percent of the calls were rated as highly necessary; 56 percent of the calls received visits. The following factors related to the preceding call increased the likelihood of a visit: (1) an early evening call; (2) a high necessity rating; (3) a classification of trauma: (4) caller at the limit of tolerance of symptoms: (5) white caller. The significance of the results and their implications are discussed.


Problems in family practice: the suicidal patient. Suicidal patients are common in family practice. Risk factors can alert the family physician to persons at unusual risk. Demographic factors, especially those revealing changing social status, recent loss, intentionality, lethality, past history, and high-risk groups, can be rapidly assessed. Viewing suicide as a final common pathway of system dysfunction at any or all major levels-biologic, psychologic, sociocultural-leads to an appreciation of the ubiquitous nature of passive or active "self murder" impulses. Persons depressed for any significant length of time for whatever reasons are potentially suicidal. Thus, the "typical" depression is suicidal but so are persons whose depression is related to drug side effects, primary illness effects, to living in a dysfunctional family system, or to normal grief. Diagnosis requires inquiry about the depth of the patient's depression pain. Treatment takes into account the aforementioned risk factors, plus interventions appropriate to the malfunctioning system.


The ultrastructural effects of acute decompression on the lung of rats: the influence of frusemide. Rats were placed in a decompression chamber and the chamber and the pressure reduced to 265 mm Hg over a period of 1 hr. Other rats were subjected to the same treatment after having been given an injection of frusemide. An ultrastructural study of the lungs of these rats showed that acute decompression of this magnitude can cause swelling and disintegration of type I alveolar epithelial cells. Although capillary endothelial cells showed only minor cytoplasmic damage many capillaries were ruptured with diapedesis of red cells. This is believed to be caused by gross capillary dilatation associated with pulmonary congestion. Pretreatment with frusemide prevented capillary rupture and reduced the degree of epithelial degeneration. However, frusemide induced swelling of capillary endothelial cells which was unassociated with acute decompression.


Acid mucins in human intestinal goblet cells. Acid mucins in goblet cells of the duodenum, jejunum and ileum of neonates and infants with cystic fibrosis (CF) and without CF were measured by scanning microdensitometry after alcian blue staining according to the protocol of McCarthy and Reid (1963) which characterises four groups of acidic mucins. In CF infants over 6 mth of age, but not in controls, there was an increase along the gut from duodenum to ileum of both weakly acidic and strongly acidic sulphomucins. In the ileum the increase was in total mucins from 6 mth previously in the same CF patients and this difference could be accounted for by an increase of sialidase-resistant mucins. The increase in sulphomucins was more marked at the tip than at the base of the villi. In CF neonates there was significant difference in the quantities of acidic mucins. The question whether the mucins of CF are chemically abnormal or merely accumulated to an abnormal extent is probably best investigated by analysis of sialidase-resistant mucins and sulphomucins of the ileum and strongly acidic sulphomucins of the duodenum and jejunum in CF infants over 6 mth of age.


Ultrastructure of human foetal and neonatal hyaline cartilage. Ultrastructural studies showed that human foetal and neonatal hyaline cartilage contained at least two types of chondrocytes. The predominant chondrocytes of chief cells were characterised by scanty, flat endoplasmic reticulum, few mitochondria, a rudimentary Golgi zone, and by the presence of clear cytoplasmic vacuoles which contained electron-dense granules and beaded filaments. The second type of cells had dark or electron-dense cytoplasm which contained numerous dilated endoplasmic reticulum and prominent Golgi zones; these cells gave rise to slender cytoplasmic processes from which the matrix vesicles were probably derived. The lacunar matrices surrounding both cell types were morphologically different reflecting some functional differences between these cells. Both types of cells participated in the formation of costochondral growth zones. Condrocytes with morphology of the chief cells proliferated, columnised, underwent hypertrophy,  and differentiated either into a compact hypertrophic cell or a swollen pale cell. The cells also became hypertrophic and gave rise to increasing numbers of dense vesicles. Calcification was first noted in the longitudinal septa inside or adjacent to matrix vesicles. The recognition of the heterogeneity of chondrocytes in human hyaline cartilage may be helpful in understanding the pathology of the chondrodysplasias.


Endocytosis and formation of macrophage polykarya: an ultrastructural study. Mononuclear phagocytes in foreign body granulomas become entrapped by the lamellipodia or enter the intracytoplasmic membranous labyrinth of macrophage polykarya. Evidence of membrane fusion was found in such instances and it is suggested that the internalisation of the mononuclear phagocyte is allied to the motility of the syncytium. This concept is also supported by the reports of other authors who emphasise the role of endocytosis in the formation of macrophage polykarya. The net effect of such a mechanism is growth of the syncytium.


The leucocoyte disappearance reaction in non-immune acute inflammation. Injection of a variety of irritants (saline, ovalbumin, compound 48/80 and powdered glass) into the rat pleural cavity induced the disappearance of pleural leucocytes during the first two hours of the reaction. This phenomenon, termed the leucocyte disappearance reaction (LDR), was suppressed by treatment with the anticoagulants heparin and warfarin. The in-vitro incubation of normal, or inflammatory pleural leucocytes resulted in the deposition of dense interconnecting meshwork of fibrin only upon addition of fibrinogen to the culture medium. It is suggested from these results that the LDR is related to the clotting system, involving leucocyte-derived enzyme(s) analogous to those of the clotting system (e.g., tissue thromboplastin), which convert fibrogen to fibrin, resulting in cell-trapping and subsequent "disappearance" of pleural leuococytes. Similarities were observed betweeen the LDR in non-immune inflammation and the macrophage disappearance reaction of cell-mediated immunity. The significance of these phenomena in the inflammatory process, both immune and non-immune, is discussed.


[Breast check-up and high risk factors in breast cancer screening (author's transl)]. Between 1966 and 1977, 142 breast cancers have been discovered in 85 250 women among 120,000 routine clinical examinations. Comparison between the information given by mammography alone compared with the panel of specialized techniques (cytology, thermography, mammography, echography) shows superiority for the latter. Complete breast check-ups have doubled the percentage of cancers discovered in the women that were examined (from 0,8% to 1,8%). A family history of breast cancer does not help in the discovery of cancer, except in cases of mammary changes noticed by the patient herself. However, these women have to be checked regularly, because the number of cancers diagnosed does not decrease at subsequent visits. The generalized practice of self-examination and of less mutilating treatment should encourage patients to consult earlier, as they have less fear that mutilating surgery will be performed.


[Present day development of the Paris System. First part: coplanar patterns and those called "in squares" (author's transl)]. The Paris System is a previsional dosimetric system which facilitates the work of curietherapists working with iridium 192 wires. Introduced about twelve years ago for radio-active implantations in one linear plane, this system has now become more generalized for implantations in two or more linear planes. After a brief review of the principles on which the Paris System is based, and introducing the new definitions arising from its development, the authors present the results of a systematic computerized study. The results obtained were used to establish, among others, the simple relationships between the geometric implantation data (disposition, length, number, separation... of the radioactive lines) and the dimensions of the volume treated, for the coplanar patterns and those called "in squares",


The relation of rheumatoid arthritis to hypogammaglobulinemia. The following hypothesis is proposed. Hypogammaglobulinemia is produced by an excess of normal 7S complement-fixing anti-immunoglobulin which is specific for "altered" Fc portions of immunoglobulin. This is present lesser amount in normal blood, is not reactive with the native immunoglobulins of blood, and plays a part in controlling the normal level of antibody production. When it is present in excess, some patients respond by producing also an excess of normal non-complement-fixing 7S anti-immunoglobulin of the same specificity as well as classical 19S IgM rheumatoid factor. These latter agents successfully prevent the hypogammaglobulinemia but they produce "side effects". These "side effects" constitute classical rheumatoid arthritis.


Studies into the occurrence of soluble antigen-antibody complexes in disease. VIII. Fractionation of rheumatoid samples containing immune complex-like material. Fractionation studies were performed of an immune complex-like material measured in rheumatoid synovial fluid using a biological assay based on histamine release from guinea pig lung. This material was eluted from Sepharose 6B in the excluded fraction, from DEAE cellulose predominantly in the third buffer, and from polyacrylamide electrophoresis in a cathodal position. Heterogeneity of the complexes was evident on iso-electric focussing, with one peak of activity at a pH of 4.2--5.6 and the other at pH 6.8--8.6. Purification of the material was limited by a loss of activity which occurred on using three fractionation procedure in sequence. This instability was avoided in the case of Sepharose 6B by adding 0.1% bovine or rabbit albumin to the elution buffer.


Inter-relationships of laboratory variables in rheumatoid arthritis. The inter-relationships of 25 variables in 140 rheumatoid patients were studied. One group, consisting mainly of hematologic variables, was differentiated on the basis of its high correlation with disease activity. The second group consisted of CRP and rheumatoid factor, which had highly significant positive correlations with each other. The third group consisted of serum aspartate aminotransferase, serum creatinine, antistreptolysin titre and blood eosinophils. These relationships are discussed.


Candida tropicalis arthritis - assessment of amphotericin B therapy. A 28 year old male heroin addict developed Candida tropicalis infection of the knee joint in association with candidemia. Assessment of amphotericin B therapy was facilitated by the determination of serum and synovial fluid amphotericin B concentrations using a radiometric bio-assay method. The results indicate that adequate synovial fluid drug levels were achieved with intravenous systemic therapy.


Rheumatoid arthritis with a normal erythroycte sedimentation rate. A persistently normal erythrocyte sedimentation rate in the presence of active synovitis was found in 17 out of 500 patients with rheumatoid arthritis. These were compared with 17 age- and sex-matched random rheumatoid arthritis controls. The patients were mostly female (15/17) and seronegative (12/17). Other acute phase reactants were measured in some cases and were also normal. Athough erosion and deformity occurred in more than half, significant involvement was restricted to the small, peripheral joints. The limited mass of inflamed synovial tissue may account for the absence of an acute phase response.


Cogan's syndrome with arthritis and aortic insufficiency. A young man with nonsyphilitic keratitis and vestibulo-auditory dysfunction (Cogan's syndrome) and a recurrent febrile illness has been observed over a 10 year period. The systemic manifestations have included a splenic abscess vasculitis, pericarditis, aortic valvulitis, and mono-articular inflammatory arthritis.


Cardiopulmonary manifestations of systemic lupus erythematosus. Nine of 17 patients (52%) with systemic lupus erythematosus participating in a controlled prospective study were found to have maximal mid-expiratory flow rates suggestive of obstructive or obstructive and restrictive pulmonary disease. Cigarette smokers with systemic lupus erythematosus had significantly lower (P less than .05) maximal mid-expiratory flow rates than either non-smokers with systemic lupus erythematosus or smokers with degenerative joint disease. By echocardiography, pericardial thickening was seen in six of 17 patients (41%) and mitral valvular abnormalities were noted in four of 17 (24%) patients with systemic lupus erythematosus. Echocardiography was found to be an effective means of detecting pericardial disease in patients with systemic lupus erythematosus.


The CREST syndrome variant of scleroderma in a mother-daughter pair. The CREST syndrome variant of scleroderma was discovered in a 48 year old woman whose mother, age 70, also had CREST syndrome characterized by extensive skin involvement. HLA  typing of the two patients, as well as the two asymptomatic daughters of the proband, revealed the A11-Bw22 haplotype. Review of the family relationships revealed two first cousin marriages. This is the fifth case report of vertical transmission of PSS or its variants, and the first report of this HLA type in such patients. An unusual feature of the illness in the mother was a large perforation of the nasal septum of the type usually associated with Wegener's disease.


Prevalence of sacroiliitis and ankylosing spondylitis in psoriasis patients. The presence of sacroiliitis was studied in 100 consecutive psoriatic patients, 62 males and 38 females. In this selected population 14 patients (14%) had grade 2 to 4 sacroiliitis, in seven associated with clinical findings of ankylosing spondylitis (AS). Sacroiliac joint involvement was significantly related to more severe cutaneous and ungual psoriatic disease. Sacroiliitis was observed in nine males and five females but clinical AS predominated in the males, six males and one female. No difference in distribution was observed by sex, onset age, or duration of psoriasis when patients with and without sacroiliitis were compared. However, patients with sacroiliitis more frequently had peripheral joint involvement.


Behçet's disease: a case with hemoptysis, pseudotumor cerbri, and arteritis. A case of Behçet's diease with massive hemoptyis is described. The hemoptysis occurred during disease exacerbation with extensive oropharyngeal and laryngeal ulcerations, and responded to high-dose corticosteroid therapy. Other unusual manifestations included a light sensitive dermatitis, pseudotumor cerebri, and right bundle branch block. Also, arteritis and subsequent occlusion of the femoral and popliteal arteries occurred. Although uncommon, pumonary involvement may be life-threatening and should be treated with corticosteroids. Both pulmonary and neurologic involvement tend to occur with active aphthosis and respond to corticosteroids. This case underscores the protean nature of the organ system involvement in Behçet's disease and supports the concept that vasculitis is a primary pathophysiologic event.


Bone ankylosis. a rare manifestation of gout. A 44 year old man with neglected tophaceous gout developed progressive bony and fibrous ankylosis of the lower leg articulations extending from a toe through the ankle, in spite of three years of otherwise successful therapy.


Impairments of glycogen and drug metabolisms by chloropeptide, a hepatotoxic mycotoxin of Penicillium islandicum sopp. Administration of chloropeptide, a hepatotoxic cyclic pentapeptide of Penicillium islandicum Sopp, caused a marked decline in the hepatic glycogen synthetase activity with concomitant increase in NADP-dependent G-6-P dehydrogenase activity. The microsomalmixed function oxygenases were also reduced in the liver of rats following chloropeptide treatment.


Potentiation of toluene toxicity by hepatic enzyme inhibition in mice. The toxicity of toluene was evaluated in male mice by the narcotic effects and lethality. The i.p. LD50 of toluene for male mice was calculated to be 1.15g/kg according to the method of Weil. Pretreatment with phenobarbital, produced a marked decrease in the number of sleep induced by a test dose (0.96 g/kg i.p.) of toluene. By contrast, pretreatment with a variety of hepatic enzyme inhibitors such as CCl4, SKF 525A, pyrazole and cyanamide was found to produce an increase in the number of sleep and death, accompanied with the prolongation of sleeping time. The present results suggest that the toxicity of toluene is enhanced by the inhibition of any step of metabolism.


Protein-mediated transfer of DDT to adipose tissue. The in vitro transfer of DDT through various media to rat adipose tissue was examined. About 15% of DDT was transferred through Krebs-Ringer solution to the tissue slices after 2-hr incubation at 37 degrees C. The addition of albumin accelerated the transfer of DDT proportionally to the concentrations up to 1.0%. Hemoglobin and gamma-globulin showed a lesser accelerating activity, but protamine, trypsin and pepsin showed little or no effect. When the Krebs-Ringer solution was replaced by triglyceride suspensions, only a small amount of DDT was taken up by the adipose tissue. Ffflux experiments revealed that the adipose tissue scarcely released DDT into albumin solution or triglyceride suspensions. The results suggest that lipid may play an important role as a storage site of DDT and that albumin may behave as a carrier system.


Effects of heavy metal ions on the electrical properties of mucous epithelial cells in the newt stomach. Zinc ion, Cu2+ and Cd2+ at 10(-4) M or lower concentrations caused a marked decrease in the membrane potential of mucous epithelial cells in the isolated newt stomach without appreciable change in the effective membrane resistance and electrical coupling ratio. Cysteine and acetylpenicillamine antagonized the action of these ions. These ions as well as ouabain blocked the hyperpolarization of membrane due to an increase in intracellular Na+. The effect of the ions on the relationship between external K+-concentration and membrane potential was similar to that of ouabain. Discussion is made on the possibility that the ions inhibit active ion transport by combining with a SH-group of the cell membrane and, thus, bring about a decrease in membrane potential.


On the metabolic detoxication of 4-iodothymol in man. The detoxication of 4-iodothymol was investigated in man. A new metabolite was isolated as a crystalline derivative from 4-iodothymol medicated urine and identified as 4-iodothymol glucuronide as a methyl acetyl derivative. The deiodinated metabolite of 4-iodothymol, thymoquinone, was also recognized in a very small amount in 4-iodothymol medicated urine.


[Neurofilament hypertrophy induced in the rabbit spinal cord after intracisternal injection of aluminum chloride (author's transl)]. The cytological changes induced in the spinal cords of rabbits by intracisternal injections of aluminum chloride have been studied at the light- and electron-microscopic level for examining the progress of the 100 A filament hypertrophy. A moderate increase in the number of the 100 A filament was detected in perikarya of neurones in anterior, lateral, and dorsal horns 4--5 days after aluminum injection. Tangles of 100 A filament were seen in perikarya and proximal parts of dendrites in these neurones 1--2 weeks after the administration of aluminum. In this stage the tangles were detected as large clear spots at light-microscopic level. The 100 A filament bundles developed in between clumps of densely aggregated endoplasmic reticulum keeping close contact with this and squeezing other organella out. In high resolution studies the 100 A filament closely resembled the neurofilament in normal neurones in size and appearances and was seen to be connected with each other via side arms coming out from its wall. Relations between the endoplasmic reticulum and the 100 A filament hypertrophy was discussed referring to intoxicating effects of aluminum compounds on the nucleus of the neuron.


[Subacute toxicity test of AMI-U-II, a new amino acid solution for renal failure (author's transl)]. Subacute toxicity test of AMI-U-II was carried out using male and female JCL:SD rats. The animals were given intravenously AMI-U-II (80, 40 and 20ml/kg) or reference solution (80 and 40ml/kg) for five weeks. Tachypnea, depression of spontaneous activity, blepharoptosis and edema of face were observed in rats treated with AMI-U-II or reference solution at highest dose. Food consumptions and gaining body weight were slightly reduced in male of these animals, but water intakes were increased in both sexes. Autopsies of the animals which died during five weeks showed pulmonary congestion and/or edema, ascites and pleural effusion. Microscopically, hydropic degeneration of liver cells and dilation of renal tubules and Bowman's capsules were shown. It seems likely that most of these findings were caused by hypervolumic administration of amino acid solution.


Rifampicin in the treatment of tuberculous meningitis. Seventy-one patients diagnosed to have tuberculous meningitis were treated with isoniazid, streptomycin plus either rifampicin (36 patients or ethambutol (35 patients). Results of therapy were identical in both treatment-groups (approximately 50 per cent mortality). Rifampicin appears to be as effective as ethambutol in the treatment of this infection.


Attitudes of Sudanese urban and rural population to mental illness. A systematic sample of 200 Sudanese individuals (100 males and 100 females) attending 4 health centres (2 urban and 2 rural) were interviewed, using standardized vignettes and a structured questionnaire, to explore their attitudes to mental illness and the mentally ill people. Information obtained from 183 (91.5%) of the respondents was analysed. Serious mental illness, represented by schizophrenia, was recognized as such by 76 per cent of both the urban and rural population and psychiatric treatment was suggested for it by more than half of them. About one-third of the respondents though of depression as just over-worry. The least recognition, as mental illness, was for alcoholism especially in the rural areas. More rural people than urban opted for religious healing as a method of treatment. Religious factors and people's concept of mental illness seem to influence their tolerance of deviant behaviour.


Isolation of Vibrio parahaemolyticus from Persian Gulf. We have been able to establish, for the first time, the presence of V. parahaemolyticus in the waters of the Persian Gulf. Samples were collected from water, fish, shrimp, sand and sea vegetation around various coastal towns of the Persian Gulf during November 1977--April 1978. Various biochemical tests have confirmed the identification of an isolate from one of the water samples as V. parahaemolyticus.


An experimental study on the role of coronary collateral development in preservation and improvement of contractile force in the ischemic myocardium. Attempts have been made to demonstrate that the presence of coronary collateral development can preserve and improve the regional contractile force in the ischemic myocardium. Studies were carried out in dogs with developed collaterals and in control dogs. Contractile force was measured with strain-gauge sutured onto the ischemic and the border areas of the myocardium. After ligation of the anterior descending coronary artery, an augmentation of coronary collateral flow via the left circumflex artery was carried out by means of infusion pump up to two and a half fold level. In dogs with developed collaterals, contractile force decreased by 57.6 +/- 11.1% (p less than 0.01) in the ischemic area and 27.2 +/- 5.4% (p less than 0.01) in the border area after coronary ligation. The marked recovery of contractile force up to 63.5 +/- 20.4% (p less than 0.05) of control level in the ischemic area and 70.4 +/- 20.4% (p less than 0.01) in the border area was observed by the augmentation. In control dogs, however, contractile force decreased by 93.2 +/- 17.2% (p less than 0.01) in the ischemic area and 24.6 +/- 5.5% (p less than 0.01) in the border area after the ligation. The augmentation of coronary flow did not recover the depressed contractile force in both areas. These results indicated that the beneficial effect was observed only when the ischemic area was supplied by developed collaterals and closely correlated with the extent of coronary collateral development.


The effects of diltiazem hydrochloride on the cardiac conduction: a clinical study of His bundle electrogram. The effects of diltiazem hydrochloride, a new Ca++ antagonistic antianginal drug, on the cardiac conductivity were evaluted in 10 patients with atherosclerotic heart disease. His bundle electrograms were obtained by catheter technique. R-R, A-H and H-V intervals were measured before and after intravenous administration of diltiazem. Diltiazem acted towards significant prolongation in A-H interval. R-R and H-V intervals were insensitive to diltiazem. Diltiazem did not interrupt the cardiac effects of atropine when A-H prolongation was still observed in the subject pretreated with atropine. In short, diltiazem depresses the conductivity of the atrioventricular node selectively and this is not mediated through vagus tone change. This Ca++ antagonist induced A-H prolongation strongly suggests the contribution of Ca++ current on the generation of atrioventricular nodal action potential in humans.


A case of myocarditis caused by Mycoplasma pneumoniae. A 16-year-old girl with myocarditis and hepatitis in the course of mycoplasma pneumoniae infection was reported. She had fever and coughed for ten days prior to admission. At the time of admission infiltrations of the left lower lung field were revealed on the chest X-ray films. The ESR was elevated and CRP+6. There were no leukocytosis and anemia, but S-GOT, S-GPT and LDH were moderately increased. On the 11th day of admission VPC in bigeminy appeared and the third sound was heard. Subsequently biphastic and inverted T waves in leads V2 and V3 and flattening of T waves in leads II and aVF appeared. At the same time, the cardiac shadow was enlarged. Antibody titer to mycoplasma pneumoniae increased to more than 1:640 two weeks after admission and then it decreased gradually. The cold agglutinin test was 1:64 on the 8th day of the disease and then it became normal. ASO, antibodies to DNA and immunoglobulins were normal; ANA, Coombs test and LE test were negative. The abnormal ECG-findings were normalized three months later.


Isolated stenosis of the pulmonary artery branches--an autopsy case with review of the literatures. A necropsy of a 21-year-old male with isolated stenosis of the pulmonary artery was presented. The pulmonary artery branches showed various fibrous thickening including atheromata and mural thrombi. Those findings appeared mostly in elastic and large muscular pulmonary arteries as well as in medium sized arteries, while the small arteries and arterioles remained in slight changes. Systemic large elastic arteries seemed to be small and showed atheromatous lesions similar to those of the main pulmonary arteries. Moreover, eight autopsy cases with pulmonary artery stenosis from the literature were shortly reviewed and some pathological findings as an aid for angiographic cardiovascular studies were discussed.


IgG.antibody-dependent cell-mediated cytotoxicity against cultured liver cells in patients with chronic active hepatitis. Antibody-dependent cell-mediated cytotoxicity was studied in patients with chronic active hepatitis (CAH) and normal controls (N) using a non-immune allogeneic lymphocytes and cultured rat parenchymal cells (Coon cells). Normal non sheep erythrocyte rosette forming cells (non E-RFC) alone showed a mild cytotoxicity to Coon cells in the absence of human sera (21.5 +/- 1.5) and in the presence of N sera (26.0) +/- 1.5) and in the presence of N sera (29.0) +/- 5.1), however, preincubation of Coon cells with CAH sera significantly augmented the cytotoxic activity of normal non E-RFC (49.4 +/- 8.7). This was blocked by aggregated IgG, anti-IgG and anti-IgG/Fc. CAH sera absorbed with Coon cells or rat hepatocyte-surface membranes failed to induce significant target cell lysis. In the absence of normal non E-RFC, N and CAH sera alone did not provoke cytotoxicity to Coon cells. Using membrane immunofluorescence method membrane-fixed IgG on target Coon cells was demonstrated in 7 of 10 sera from CAH. It seems likely that the IgG.antibody-dependent cell-mediated cytotoxicity may play a role in pathogenesis of CAH.


Toxicological approaches to the metabolities of fusaria. XII. Fate and distribution of T-2 toxin in mice. Tissue distribution, excretion and fate of T-2 toxin, a potent cytotoxic mycotoxin of Fusarium species, were examined in mice and rats by using tracer method as well as chromatographic technique. Orally administered T-2 toxin to mice and rats was rapidly eliminated into the feces and urine with a ratio of 4--5:1. In mice, the toxin distrubited in the liver, kidney and other organs without showing a specific accumulation. In rats, the toxin was excreted into the feces partly as HT-2 toxin, 8-hydroxydiacetoxyscirpenol (neosolaniol) and three unidentified metabolites. Toxicological significance of the hepatic and derm floral carboxyesterase was discussed in regard to the specific deacetylation of T-2 toxin.


Evaluation of the intrathoracic injection method for screening of filaricides. Intrathoracic injection technique was utilized to examine its value in screening of antifilaria drugs in Litomosoides carinii in the cotton rat, Sigmodon hispidus, using existing filaricides such as diethylcarbamazine, Mel W. metrifonate, suramin, arsenics and antimonials. Diethylcarbamazine at a dose of 100 mg/kg for 5 days given intrathoracically caused marked decrease of more than 95% of the microfilaria count in the blood 1 week after the initial injection which is the same effect as was observed by intraperitoneal injection. Whereas adult worms in the pleural cavity were not affected. Mel W at a dose of 10 mg/kg for 5 days killed all adults but had no effect on microfilaria density in the blood. When it was given intraperitoneally, even a larger dose of 50 mg/kg did not affect adults in the pleural cavity. In cotton rats tolerating suramin at 80 and 40 mg/kg for 5 days, adult worms were intact. However, a reduced dose of 20 mg/kg and 10 mg/kg given weekly for 6 weeks showed remarkable macrofilaricidal activity. Thus the slow action of suramin was reproduced by this method. There was no significant change in the microfilaria density in the blood. Aresenics such as Mapharsemin and Neo Neo Arsemin, and antimonials such as stibophen and tartar emetic were shown to have macrofilaricidal activity by intrathoracic injection with no effect on microfilaria density in the blood within a week. Effect of existing filaricides by this method showed remarkable coincidence with their action Wuchereria bancrofti.


Protease-induced experimental emphysema: the relationship between elastolytic activity and emphysema induction. Induction of experimental emphysema by protease was performed with several proteases both in vivo and in vitro to compare the ability of inducing emphysematous change and their elastolytic activity. The following results were obtained. 1) Only elastase and papain have emphysema inducing capacity. Emphysematous changes induced by elastase in vitro were dose dependent. But papain has no genuine elastolytic activity. Nature of the emphysema-inducing capacity of papain remains obscure. 2) Advantages of using the isolated lung for experiment were discussed, especially for the small dose required for enzyme-instillation. Moreover, there is no interferencey by endogeneous enzyme or protease-inhibitors. 3) Guinea pig is useful for the experiemnt of emphysema-induction both in vivo and in vitro. Minimal requirement of elastase is 50 microgram in vitro and 250 microgram for in vivo experiment.


[Effects of hippocampal lesions on punishment training in rats (author's transl)]. The present experiment was designed to determine the effect of hippocampal lesions on punishment training and to evaluate the spatial cognition hypothesis. Hippocampectomized and operated control rats were each divided into runway and goal box punishment groups. Food motivated animals were tested in the elevated straight alley on 10 acquisition days and 1 punishment day. Start box latency, runway running time, goal box entry time and goal box time were measured. The controls spent longer time in all measures on the postashock trial. The hippocampals showed no hesitation in the start box latency on the postshock trial, but they spent longer time in other measures. The location of punishment had significant effects only on the start box latency of the controls. These results support the spatial cognition hypothesis.


[A developmental study of visual-motor function in cerebral palsied children (author's transl)]. The purpose of this study was to investigate whether the disorder in the visual-motor function of cerebral palsied children was affected by specific defects or by the distortion in perceptual motor system or by mere developmental lag in the function of recognition and reconstruction using the method of multiple choice and Shochi's Block Design Test (1971). The results were as follows : Cerebral palsied children's developmental lag in the function of recognition was not conspicuous but the development in the function of reconstruction was obviously inferior to that of normal children. There was the difference in reconstruction ability by children with the types of cerebral palsy and intellectual levels and design-constructive patterns. Those factors and the developmental lag cause visual-motor disorders.


[Hemispheric differences in the mode of information processing (author's transl)]. To examine hemispheric differences in the mode of information processing, 24 normal right-handed subjects were required to make same or different responses to a set of letters varying in number from two to four which were all identical or contained one letter differing from the rest. When letters were presented in the normal upright position, the left hemisphere showed serial processing wherein reaction times increased as a function of number of letters. The right hemisphere, however, showed no increase of reaction times, i.e. parallel processing. When letters were presented in the inverted upside-down position, the left hemisphere showed partial serial processing, despite letters being analyzed physically. The results suggest that visuopatial analysis doesn't always force parallel processing in the left hemisphere.


[Two kinds of apparent size distortion in the Ponzo illusion (author's transl)]. It was proposed that there are two kinds of apparent size distortion to be distinguished in the Ponzo illusion. The apparent size of the single line in the converging lines (CL) became larger as it was located nearer to the apex. However, this distortion could not predict the obtained illusion sufficiently. Hence there should be assumed another kind of size distortion in the perception of the size relation as a result of comparison of two paralle lines (PL). In this comparison process, breaking of the continuity of CL and elimination of the oblique components from CL had a slight decreasing effect on the illusion. Moreover, it was found in the various contour figures that the greater the separation of PL was, the larger the illusion was.


[Effects of a warning signal on heart rate and 17-hydroxycorticosteroid in urine (author's transl)]. The purpose of this study was to test whether a warning signal given prior to an unpleasant stimulus would influence heart rate (HR) and 17-Hydroxycorticosteroid in urine (17-OHCS). The results for HR were as follows: (a) In the anticipation period, subjects given no warning signal (NS group) showed higher degree in their HR than that of those given a warning signal (S group), (b) On at impact, a warning signal tended to decrease subjects' HR when an unpleasant stimulus was given, (c) In the recovery period, subject group indicated lower degree in their HR than that of NS group, and (d) Subject group showed more adaptation by repeated trials to an unpleasant stimulus in their HR. However, as for urinary 17-OHC, nosignificant difference was observed.


[EEG asymmetry during mental arithmetic and recalls of verbal (kanji, kana, the alphabet) and visual imageries (author's transl)]. EEG was recorded from the left and the right parietal areas in normal subjects with eyes-closed while they were engaged in mental arithmetic, recalls of verbal (Japanese kanji characters, kana syllabary, and the alphabet) and visual (scenery) imageries. Fourier analysis of EEG revealed attenuations of the alpha activity (a) in the left hemisphere during the mental arithmetic, (b) in the right hemisphere during the visual imagination of a scenery, and (c) in the both hemispheres during the kanji imagery accompanies by a stroke-count task. Results suggest that the kanji characters seem to activate cognitive processes of the both hemispheres. No attenuation of the alpha activity in the both hemispheres was seen during either the alphabet or kana syllabary imagery.


Lactate and pyruvate metabolism in isolated renal tubules of normal dogs. The kinetics of lactate and pyruvate (1 and 5 mM in each case) metabolism was studied in isolated dog renal tubules. Utilization of these two substrates and the production of glucose, pyruvate, or lactate, and alanine were determined. The rates of lactate and pyruvate utilization and of glucose production were constant during 60 min of incubation. Glucose production from pyruvate was less than that from lactate. Addition of albumin to the incubation medium greatly inhibited lactate and pyruvate utilization at both substrate concentrations. It stimulated, however, glucose production from 1 mM, but not 5 mM, lactate or pyruvate. These effects were found to be due to the presence of fatty acids in the albumin solution used. In the absence of fatty acids, glucose production represented 35 to 40% of lactate uptake, but represented less than 20% of pyruvate uptake. Fatty acids markedly enhanced the percentage of transformation of lactate and pyruvate into glucose, and that of pyruvate into lactate. Alanine represented 20% or less of lactate and pyruvate uptake. These results suggest that fatty acids have a regulatory influence on lactate and pyruvate dog kidney metabolism.


Structural-functional heterogeneity along the rabbit collecting tubule. The possibility that the rabbit collecting tubule is heterogeneous with respect to electrical and morphologic properties was examined in isolated segments. Collecting tubules from rabbits treated with desoxycorticosterone acetate were dissected from various locations within the cortex and outer medulla, and the transepithelial potential was measured. The cortical collecting tubule always displayed a negative potential. In contrast, collecting tubules dissected from the inner stripe of the outer medulla displayed a positive potential. That this change is a function of position within the kidney is supported by the observation that some tubules generated a negative potential at the superficial (cortical) end and a positive potential at the deep (medullary) end of the same tubule. Histologic evaluation of tubules after perfusion resulted in a striking correlation between the existence of a negative potential and the presence of dark cells. The results of this study provide evidence that the collecting tubule is heterogeneous with respect to active transport processes and suggest that these functional differences may correlate with morphologic differences between the cortical segment and the medullary segment of the collecting tubule.


Radioimmunologic method for detection of antitubular basement membrane antibodies. A radioimmunoassay for detection of antitubular basement membrane (TBM) antibodies was set up using a human TBM antigen (mol wt, 70,000 daltons), purified after collagenase treatment of the insoluble membrane by preparative polyacrylamide electrophoresis, and labeled with iodine 125. Free labeled antigens were separated from those bound to immunoglobulins by a 20% polyethylene glycol (mol wt, 6,000 daltons) solution. In the presence of normal human or Brown Norway rat sera, less than 10% of the labeled antigens were precipitated. In the presence of sera or of kidney eluates from rats immunized with human TBM, the precipitation of the labeled TBM antigens reached 73%, but in the presence of sera from two patients presenting with an interstitial nephritis and linear deposits along the TBM only, up to 47% of the same antigens were precipitated. In these two cases, the anti-TBM antibodies were mainly directed against the heteropolysaccharide-containing glycopeptides isolated from TBM, that is, against the noncollagenous polypeptides of the TBM antigens. Anti-TBM antibodies were sought in the sera of 52 normal blood donors and of 11 patients presenting with glomerulonephritis and linear deposits of immunoglobulins. The average percentage (+/- 1 SD) of labeled TBM antigens precipitated in the serum of normal blood donors was 7.1 +/- 1.2. Of the patients presenting with glomerulonephritis and linear deposits along the GBM, 9 out of 11 exhibited anti-TBM antibodies by radioimmunoassay; among these 9 patients, 8 also displayed linear deposits of IgG along the TBM. Absorption of anti-TBM and anti-GMB antibodies with particulate TBM or GBM, with both types of glycopeptides isolated from GBM or TBM, indicated that the anti-TBM antibodies were directed against the noncollagenous polypeptides of TBM but that the anti-GBM antibodies mainly reacted with the collagenous polypeptides of TBM and GBM. Finally, it was found that the sera of 2 patients out of 15 presenting with lupus nephritis contained a significant anti-TBM-binding activity, mainly directed against the noncollagenous material of TBM.


Antibody responses to influenza vaccination in patients with chronic renal failure. A bivalent type A inactivated influenza virus vaccine containing both sets of H3N2 and Hsw1N1 antigens was given to patients with chronic renal failure and to control subjects. The patients were divided into an azotemic group in whom dialysis was not yet required and a hemodialyzed group. Hemagglutination-inhibition (HI) antibody responses were measured at time intervals of 1,3, and 4 weeks after vaccination. We found that the mean postvaccination HI titers against both sets of antigens in the patients as a group did not differ significantly from those found in the control subjects as a group. The proportion of responders showing a fourfold or greater increase in post-vaccination antibody responses over prevaccination antibody values for either set of antigens was similar in both groups of patients and the group of control subjects. In general, an inverse correlation was found between prevaccination antibody levels and postvaccination antibody responses in both patients and control subjects. The only exception to this trend was the response of two of the azotemic patients to the H3N2 antigens who failed to respond despite low prevaccination antibody levels. These were the patients with the highest serum creatinine values.


Independence of normal phenotypic properties and cell surface receptor mobility in variant cell lines. We report the use of three classes of variants from the long-established, malignantly transformed LM cell line to demonstrate that the apparent mobility of cell surface receptors need not be dependent on the expression of the transformed phenotype in vitro.


Glycolipids as indicators of tumorigenesis. Hyperplastic liver nodules and hepatocellular carcinomas were induced in rats by oral administration of the carcinogen N-2-fluorenylacetamide. Neoplastic tissue was compared with control, fetal, neonatal, and precancerous liver tissues. The development of the tumors was slow, such that temporal changes in the biochemical and morphologic development of carcinogenesis could be identified. Ganglioside sialic acid levels were elevated in all but the most poorly differentiated tumors. Experiments to monitor individual enzymes suggested that the alterations in glycolipid composition were a direct effect of alterations in biosynthetic activities. The pattern during tumorigenesis was the inverse of that during normal development. Also, ganglioside patterns showed a progressive simplification from hyperplastic nodules to well-differentiated hepatomas and through two grades of poorly differentiated hepatomas. An increase in the activity of the branchpoint enzyme of ganglioside biosynthesis preceded both a decrease in the branchpoint enzyme of the disialoganglioside pathway and a marked increase in the galactosyltranferase of GM1 formation. The results indicate that ganglioside deletions are the end result of a cascade of events in the tumorigenic transformation. The onset of ganglioside deletions but not of the cascade per se may correlate with the onset of malignancy. Glycolipid levels are elevated early in certain surrounding tissues especially in the blood. In rats bearing transplantable hepatomas, serum levels of lipid-bound sialic acid were elevated 2.5-fold. Similar results were obtained with sera of mice bearing transplantable mammary carcinomas and of cancer patients. These findings provide new emphasis for gangliosides in both cancer detection and as regulatory signals for growth and multiplication of cells.


Lipopolysaccharide effects on sensitive and resistant variant Chinese hamster ovary cell lines. Chinese hamster ovary (CHO . K1 . PRO) cell growth was inhibited by addition of a gram-negative bacterial lipopolysaccharide (LPS) to the cell culture medium. Growth inhibition began after three or four days of incubation, was dose-dependent up to a maximum at an LPS concentration of 500 microgram/ml and was accompanied by cell shape changes and enhanced cytoplasmic vacuolization. Formation of bizarre CHO . K1 . PRO cell shapes and vacuole formation were most pronounced after seven days of incubation with LPS and could be observed by light and electron microscopy. An LPS-resistant cell population was obtained by intermittent in vitro exposure to high levels of LPS; these variant cells or clones derived from them failed to display growth inhibition in the presence of LPS. A clone from the LPS-resistant variant population showed altered cell properties compared to the parental cell line which included changes in cell morphology, adhesion, and endocytosis. Parental cells was markedly density-inhibited, whereas the cariant clone exhibited considerable growth after confluency. The LPS-resistant variant cells showed a more elongated morphology than the parental line. No significant differences were observed between rates of detachment of parental and variant cells when sparse cultures of either line were removed from tissue culture dishes by ethylenediaminetetracetate (EDTA). However, at confluency approximately 100% of the variant cells versus 35% of the parental cells were removed by EDTA in one hour. Measurements of 125I-ferritin uptake by parental and variant cells showed approximately twenty-fold and twofold increases, respectively, in uptake induced by LPS when compared to untreated control cultures.


Binding of ricin A chain to rat liver ribosomes: relationship to ribosome inactivation. Ricin A chain was radioactively labeled using reductive alkylation, lactoperoxidase catalyzed iodination, and reaction with iodoacetamide or N-ethylmaleimide (NEM). The inhibition of cell-free rat liver protein synthesis by the modified A chains and the ribosome binding characteristics of each of the labeled derivatives was examined. [3H] NEW was found to quantitatively react with the A chain sulfhydryl group normally involved in a disulfide bond with the B chain in intact ricin. Labeling the protein with [3H] NEM had no effect on the in vitro inhibition of protein synthesis by the A chain. [3H] NEM-labeled A chain binds to rat liver ribosomes in a manner which is dependent on the concentrations of NaCl and Mg2+. At optimal Mg2+ concentration (5.5 mM), A chain binding to ribosomes is saturable and fully reversible either by dilution of the reaction mixture or by addition of unlabeled A chain. At 5.5 mM Mg2+, A chain was found to bind to a single site on rat liver ribosomes with a dissociation constant of 6.2 x 10(-8) M. [3H] NEM-labeled A chain did not bind to isolated 40S ribosomal subunits and bound to 60S ribosomal subunits with a 1 : 1 molar stoichiometry and a dissociation constant of 2.2 x 10(-7) M. The relationship between ribosome binding and A chain inhibition of eucaryotic protein synthesis is discussed.


Membrane effects of imidoesters in hereditary stomatocytosis. The marked increase in cation (Na+, K+) permeability that results in swollen, cup-shaped red cells in the hereditary stomatocytosis syndrome can be corrected in vitro with a bifunctional crosslinking reagent, dimethyl adipimidate (DMA). 45Ca influx in intact RBC, 45Ca efflux in red ghosts, and 45Ca retention in red ghosts are normal and not influenced by DMA. Endocytosis in resealed red ghosts is strikingly impaired but becomes normal if cells are first treated with 2 mM DMA. Protein kinase mediated phosphorylation of membrane proteins by AT32P--only 20--40% of normal control values in both shortterm (5 min) and more extended (60 min) incubations--is not improved by DMA. After reaction of 14C-DMA with stomatocytes, radiolabel is found associated with phosphatidyl serine and phosphatidyl ethanolamine and is also widely distributed among membrane proteins. Cation permeability of stomatocytes in corrected at DMA concentrations (1 mM) that result in barely detectable crosslinking of aminophospholipids or proteins, suggesting that either crosslinking of a minor component present in only small quantities or intramolecular (rather than intermolecular) crosslinking is responsible for the permeability effects. DMA, whose maximal crosslinking dimension is 7.3--9 A, is the most effective bifunctional imidoester of those tested. Shorter (dimethyl malonimidate) or longer (dimethylsuberimidate) reagents are either less effective than DMA or totally without effect.


Plasma membrane proteins and glycoproteins from Chinese hamster cells sensitive and resistant to actinomycin D. Plasma membrane proteins and glycoproteins have been isolated from Chinese hamster cells of the spontaneously transformed DC-3F parental cell line and the DC-3F/AD X line with a high level of acquired resistance to actinomycin D. Plasma membrane preparations from both cell lines band at 1.16 g/ml after isopycnic centrifugation. We present evidence to indicate differences in the leucylpeptide backbones of the antibiotic-sensitive cells and the drug-resistant DC-3F/AD X cells. In addition, there are differences in the plasma membrane glycopeptides of the two cell lines as revealed by sodium dodecyl gel electrophoresis. Drug-resistant cells synthesize a surface glycopeptide which is much larger than the major one present on the drug-sensitive cells. Both of these cell lines are devoid of 5'-nucleotidase and alkaline phosphatase activities. The role of plasma membrane protein differences in drug-resistant cells is discussed.


Transmembrane action of thrombin initiates chick cell division. Thrombin immobilized on polystyrene beads initiates DNA synthesis and cell division in quiescent cultures of chick embryo (CE) cells in serum-free medium. These thrombin beads also produce morphological changes in CE cells similar to those produced by soluble thrombin. The amount of acid-precipitable material released from 125I-thrombin-beads into the culture medium was 60-fold less than the amount of soluble thrombin required to produce an equivalent increase in cell number. Moreover, EM autoradiography of CE cells 10 h after 125I-thrombin-bead addition showed that there was no direct release and accumulation of radioactive material in the cytoplasm of these cells. These results demonstrate that thrombin action at the cell surface is sufficient to initiate proliferation of CE cells.


Resistance of the intact and reconstituted adipocyte hexose transport system to irreversible inhibition by sulfhydryl and amino reagents. Sensitivity of the adipocyte D-glucose transport system in intact plasma membranes or following solubilization and reconstruction into phospholipid vesicles to several protein-modifying reagents was investigated. When intact plasma membranes were incubated with N-ethylmaleimide (20 mM) or fluorodinitrobenzene (4 mM), D-glucose transport activity was virtually abolished. However, washing the membranes free of unreacted reagents restored transport activity, indicating that covalent interaction with the membrane did not mediate the transport inhibition. Reaction of [3H]N-ethylmaleimide with plasma membranes under similar conditions resulted in extensive labeling of all protein fractions resolved on dodecyl sulfate gels. Similarly, addition of N-ethylmaleimide to cholate-solubilized membrane protein had no effect on transport activity in artificial phospholipid vesicles reconstituted under conditions where the membrane protein was free of unreacted N-ethylmaleimide. Transport activity in plasma membranes was also inhibited by both reduced and oxidized dithiothreitol or glutathione (15 mM) in a readily reversible manner. consistent with a noncovalent mode of inhibition. Thus, the insulin-responsive adipocyte D-glucose transport system differs from the red cell hexose transport system in its remarkable insensitivity to modulation by covalent blockade of sulfhydryal or amino groups by the reagents studied.


Transmembrane interactions and the mechanisms of transport of proteins across membranes. We have made observations, by double fluorescence staining of the same cell, of the distributions of surface receptors, and of intracellular actin and myosin, on cultured normal fibroblasts and other flat cells, and on lymphocytes and other rounded cells. The binding of multivalent ligands (a lectin or specific antibodies) to a cell surface receptor on flat cells clusters the cell receptors into small patches, which line up directly over the actin- and myosin-containing stress fibers inside the cell. Similar ligands binding to rounded cells can cause their surface receptors to be collected into caps on the surface, and these caps are invariably found to be associated with concentrations of actin and myosin under the capped membrane. Although these ligand-induced surface phenomena appear to be different on flat and rounded cells, we propose that in both cases clusters of receptors become linked across the membrane to actin- and myosin-containing structures. In flat cells these structures are very long stress fibers; therefore, when clusters of receptors become linked to these fibers, the clusters are immobilized. In round cells, membrane-associated actin- and myosin-containing structures are apparently much less extensive than in flat cells; therefore, clusters of receptors linked to these structures are still mobile in the plane of the membrane. We suggest that in this case the clusters are then actively collected into a cap by an analogue of the muscle sliding filament mechanism. To explain the transmembrane linkage, we propose that actin is associated with the plasma membrane as a peripheral protein which is directly or indirectly bound to an integral protein (or proteins) X of the membrane. Individual molecules of any receptor are not bound to X, but after they are specifically clustered into patches, a patch of receptors then becomes bound to S and hence to actin/myosin.


Effects of PTH and Ca2+ on renal adenyl cyclase. The effects of calcium ion on the adenylate cyclase system was studied in isolated, renal basal-lateral plasma membranes of the rat. Bovine parathyroid hormone (bPTH) and a guanyl triphosphate analogue, Gpp(NH)p were used to stimulate cyclase activity. Under conditions of maximal stimulation, calcium ions inhibited cyclic adenosine monophosphate (cAMP) formation, the formation rate falling exponentially with the calcium concentration. Fifty percent inhibition of either bPTH- or Gpp(NH)p-stimulated activity was given by approximately 50 microns Ca++. Also the Hill coefficient for the inhibition was close to unity in both cases. The concentration of bPTH giving half-maximal stimulation of cAMP formation (1.8 x 10(-8) M) was unchanged by the presence of calcium. These data suggest that calcium acts at some point other than the initial hormone-receptor interaction, presumably decreasing the catalytic efficiency of the enzymic moiety of the membrane complex.


Bilayer balance and regulation of red cell shape changes. Discocytic human red cells undergo discocyte-echinocyte and discocyte-stomatocyte transformations under the action of a wide variety of lipid-soluble anionic and cationic agents respectively. These shape transformations are explained by the bilayer couple hypothesis of Sheetz and Singer to be the result of preferential distribution of the anionic agents in the outer half of the bilayer and the cationic agents in the inner half of the bilayer. We demonstrate that echinocytogenic effects indeed occur when the naturally occurring phospholipid lysophosphatidylcholine (LPC) is localized in the outer half of the bilayer, and stomatocytogenic effects occur when LPC is in the inner half. However, in contrast to the bilayer couple hypothesis, our results show that simple equivalent membrane surface area expansion on each layer is insufficient to maintain the discocytic shape and there exists a differential concentration effect of LPC on the two halves of the bilayer.


Production of complete and incomplete antibodies to human red blood cells in dogs and rabbits. The characteristics of the immune response of dogs and rabbits to single and multiple injections of human red blood cells were analysed. The results show that the primary and secondary response of both species are essentially the same. Following repeated stimulation with this antigen quantitative differences of the immune response between these animals were observed. The rabbit produced substantially greater amounts of IgG complete and incomplete antibody, and proportionally greater amounts of IgM antibody, whereas the dog response was seen to be inhibited.


A quantitative ultrastructural and chemical investigation of the accumulation of iodophilic polysaccharide in two cariogenic strains of Streptococcus mutans. The amount of intracellular, iodophilic, glycogen-like polysaccharide (IPS) present in cells of two strains of Streptococcus mutans at various stages of growth in a chemically defined medium was determined by quantitative electron microscopy. The results obtained were then compared with the chemically determined, iodophilic polysaccharide content of cultures. The ultrastructural method used determined the fraction of area of central longitudinal sections of individual cells occupied by stained granules, and was therefore capable of determining amounts of polysaccharide in cells starved of glucose. Although the results of the two methods showed very good quantitative correlation, the ultrastructural method allowed study of glycogen synthesis on a cellular basis, and detected some heterogeneity in amounts of IPS stored by individual cells. The ultrastructural method also permits the detection of much smaller amounts of stored IPS than does the chemical method.


Pathogenicity and pathology of Jos virus infection in mice and tissue culture. Jos virus was pathogenic to suckling mice infected by intracerebral (i.c.), intraperitoneal (i.p.), subcutaneous (s.c.), and oral (per os) routes. However, consistent fatality was only obtained in adult mice infected intracerebrally. Suckling mice inoculated by the i.c. route developed viraemia within 24 h post infection, and virions were rapidly disseminated into all visceral organs. High infectivity titres found in organs such as liver and lung as compared to blood indicated that virus multiplication took place in them. Pathological studies in infected suckling mice showed acute cell necrosis in the liver, lymph nodes, bone marrow and spleen. Other organs, including the brain, were unaffected. Secondary cell culture did not readily support growth of the virus and there was no evidence of cytopathic effect or virus multiplication in Vero and BHK-21 cell culture after three passages.


Some physiological studies on Penicillium expansum Link. Penicillium expansum Link showed excellent/good growth on all carbohydrates except ribose, lactose, sorbose and sucrose, which induced fair or poor growth. It made excellent growth on mannitol and dulcitol while the rest of the carbon sources were either poorly utilized or not utilized. Sorbose neither inhibited nor promoted growth of the pathogen in the presence of good carbon sources. The pathogen showed fair/poor growth on cystine, phenylalanine, histidine, mono-HCl, histidine di-HCl, ammonium sulphate, ammonium nitrate, ammonium chloride, ammonium phosphate, and urea, while the rest of the nitrogen sources induced excellent/good growth. However, there was no growth on potassium nitrite. Cystine inhibited the growth of the pathogen to some extent in the presence of good amino acids, i.e. serine proline, methionine, arginine mono-HCl, tyrosine and tryptophane, and not at all were inhibited in the presence of the remaining amino acids.


Biological autoxidation. II. Cholesterol esters as inert barrier antioxidants. Self-assembly of porous membrane sacs. An hypothesis. The antioxidation defenses recognized thus far appear too weak. Needed are inert barriers to encapsulate foci of activated oxygen (FAOs) and contain their spreading. These capsules must: 1. self-assemble nonenzymatically and spontaneously in face of adversity; 2. resist oxidation and dissolution in water; and 3. be moderately fluid and elastic enough to withstand flexing by tissues. Evidence shows activated oxygen: a. is produced by common cholesterolester (CE)-raising agents; b. boosts accumulation of CEs; and c. splits low-density lipoproteins (LDL), thus releasing CE-rich coalescence-prone lipid micelles. I am proposing that CEs, combined with polar lipids, are uniquely suited to form inert-lipid antioxidation barriers (ILABs). Porous ILAB capsules self-assemble from lipid micelles released by oxidatively degraded LDL. The capsules are thermodynamically unstable but elastic, durable and capable of self-repair through oxidation of ambient LDL. All capsules tend to contract into spheres. Enclosed needle-like "foreign bodies", such as asbestos, puncture the contracting capsules. Hence the odd bulbous architecture of asbestos bodies. ILABs protect from--and their failure initiates and promotes--carcinogenesis and atherosclerosis. ILABs may be mediators of membrane biogenesis. The loss of arterial flexibility in atherosclerosis protects ILAB capsules from breakage.


A hypothesis concerning the structural relationship between memory and emotion. Limbic and temporal lobe structures have long been implicated as the anatomic substratum for emotion. Recent investigations have localized the processing of short term memory to a virtually identical anatomical locus. It would seem logical that the same structures subserve both functions if one assumes that the strength of the memory trace is dependent on the emotional value of the information for the organism. Under this hypothesis, the hippocampus emotionally weights sensory data as a preliminary to incorporation of such material into long term memory. It is not necessary, nor is it likely, that long term memory reside within the temporal lobe. Previous associations stored in remote memory can be accessed by the temporal lobe and these may serve to modulate the weighting of the present stimulus. Some of the functions of the temporal lobe can be modeled by a simple electronic filter; the addition of a single feedback loop to this system can account for other properties inherent in the hypothesis. The anatomical connections of the system would permit it to be an early and efficient repressor, in the pyschodynamic sense.


The olive and central control of blood pressure. This evidence, then, would suggest that the olive becomes active when, with a change in the motor task, the motor performance is required to adjust to the new circumstance. From Smith's experiments on blood pressure, it is extrapolated that the olive becomes active when there is a change in the physiological state, and the autonomic performance is required to adjust to the new circumstance. The important point is that the olive does have an experimental effect on blood pressure, and that it is a specific, though indirect, effect acting through the carotid sinus reflex. This effect appears similar to the effect on motor control which is specific for motor learning, and appears to act indirectly through the cerebellum. The hypothesis is being put forward because of the importance of blood pressure abnormalities, and the possible therpeutic benefits to be derived from an understanding of the central control of blood pressure. It is hoped that in pursuing this elusive control system one shall not overlook the olive.


The relationship of hormones to arterial glycosaminoglycans and atherosclerosis. Glycosaminoglycan fractions were measured in representative large and medium sized arteries of normal, hypophysectomized and hormone treated young beagles. Hyaluronate, heparan sulphate, dermatan sulphate and the isomeric chondroitin sulphates were determined in the aortic arch, thoracic and abdominal segments, in the external iliac, superior mesenteric, renal, common carotid and coronary arteries. The hormones used for replacement therapy of hypophysectomized animals were growth hormone, thyroxine, cortisone and the sex hormones testosterone, estrogen and progesterone. The sensitivity to an individual hormone was found to differ in various segments of the arterial tree; the thoracic and abdominal aorta were most responsive but renal and superior mesenteric arteries were relatively inert. The hypothesis is advanced that arteries with a GAG metabolism highly sensitive to hormones are more prone to develop atherosclerosis than arteries that have a limited sensitivity to alterations in endocrine balance.


Role of platelet factors in the growth of cells in culture. The studies reported suggest that the principal mitogen(s) present in sera responsible for the proliferation of diploid cells in culture is derived from the physiologic response of platelet adherence, aggregation, and release upon their exposure to factors present in serum, such as thrombin, or in tissue, such as collagen. Since it is impossible to make whole blood serum without platelet release, all sera contain platelet mitogenic factor(s). In contrast, serum made from platelet-free plasma lacks mitogenic activity and permits maintenance of cells in culture in a quiescent state for long periods if the cells are routinely fed. The platelet factor(s) appears to be a heat-stable, basic polypeptide or protein, that upon exposure to the cells recruits them into the cell cycle, DNA synthesis, and mitosis. The factor(s) has been shown to act not only in cell culture but in vivo as well. Maintaining cells in a culture medium containing platelet-free, plasma-derived serum may be more analogous to the quiescence of adult cells in vivo, since quiescent cells in adult tissues are normally exposed to interstitial fluid that is probably more like a filtrate of plasma or lymph rather than to whole blood serum. In contrast, growth of cells in a culture medium containing whole blood serum would be more analogous to the pathologic situation that occurs during tissue injury accompanied by hemorrhage.


Role of bovine albumin in a serum-free culture medium and its application. Our experiments showed that fatty acids bound to bovine serum albumin V (BSA) played an important role in the growth of cells in primary culture. A serum-free culture medium containing BSA could support continuous growth of various normal and malignant cells from primary culture, and the amount of BSA in the medium could be reduced by supplementation with minute amounts of basic high polymers.


The somatomedin group of peptide growth factors. The somatomedins, a group of serum growth factors, have in common the ability to stimulate proteoglycan synthesis in cartilage. All are growth hormone dependent and have insulinomimetic properties in extraskeletal tissues. All stimulate DNA synthesis and mitosis in a variety of cell lines. Somatomedin-A and multiplication-stimulating activity are neutral peptides of 7,000--10,000 daltons, whereas nonsuppressible insulin-like activity and somatomedin-C are basic peptides of 5,700--7,000 daltons. Whether some of these substances are identical remains to be established. By use of a highly specific  radioimmunoassay for somatomedin-C and less specific radioreceptor assays for insulin-like activity and somatomedin-C, evidence has been obtained for the existence of two major classes of somatomedins: neutral somatomedin, which is more insulin-like, and basic somatomedin, which is more rigorously growth hormone dependent, and which, in some systems, is s more powerful stimulant of growth.


Multiplication-stimulating activity (MSA): a somatomedin-like polypeptide from cultured rat liver cells. Multiplication-stimulating activity (MSA) has been purified to homogeneity with the use of Dowex chromatography. Sephadex gel filtration, and preparative disc gel electrophoresis. A molecular weight of 8,700 was determined for both native and reduced and alkylated MSA by chromatography on a 6% agarose column in 6 M guanidine HCl. Amino acid analysis of performic acid-oxidized MSA revealed 2--3 cysteic acid residues, and reduction and alkylation resulted in loss of biologic residues, and reduction and alkylation resulted in loss of biologic activity. These results suggested the presence on one intrachain disulfide bond, which is required for biologic activity. Specific receptors for MSA have been identified on chick embryo fibroblasts, human skin fibroblasts, a rat liver cell line, and purified rat liver plasma membranes. The closely related peptides, acid ethanol-soluble nonsuppressible insulin-like activity (NSILA-S) and somatomedin-A, competed for MSA tracer binding to these MSA receptors, whereas unrelated peptides did not compete. MSA receptors were divided into two types based on their reactivity with insulin and proinsulin. Insulin and proinsulin competed potently for MSA binding to chick embryo and human skin fibroblasts (type II) but did not compete for MSA tracer binding to rat liver cells and purified rat liver membranes (type I). In chick embryo fibroblasts, the concentrations of MSA, insulin, proinsulin, and somatomedin-A, which inhibited [125I]MSA binding by 50%, also gave half-maximal stimulation of DNA synthesis, consistent with the MSA receptor being a mediator of DNA synthesis. MSA tracer has also been shown to bind specifically to rat serum proteins. The MSA binding profile on Sephadex G-200 was shown to be growth hormone dependent. Since the serum half-life of somatomedin activity in the rat was also growth hormone dependent, these results suggest that growth hormone induces the binding protein for somatomedin and thereby governs the half-life of somatomedin. Finally, the ability of two rat liver cells lines to multiply in serum-free medium did not depend upon the level of MSA in the conditioned medium.


Cell culture for the study of epithelial cells. Keratinocytes depend on the support of fibroblasts or fibroblast products to grow from single cells into colonies. The essentials of a human stratified squamous epithelium can be constructed from single human epidermal keratinocytes and lethally irradiated fibroblasts. Established lines of mouse keratinocytes obtained from teratomas have many of the same properties. In this way it is possible to study many aspects of this epithelial tissue or organ under essentially cell culture conditions.


Induction of differentiation in mouse neuroblastoma cells. In the presence of 1--2% dimethyl sulfoxide (DMSO), mouse neuroblastoma cells are induced to differentiate morphologically as well as electrically. In addition, treatment of neurolbastoma cells with 2% DMSO results in a marked increase in the veratridine-activated K+ or Rb+ efflux. At 4% DMSO, neurite outgrowth is completely repressed and electrical activity is poorly developed. However, at this concentration, the cells have a relatively high resting potential which suggested that membrane components determining passive and active permeability properties are not necessarily under coordinated control. Induction of differentiation by 2% DMSO is also accompanied by an increase in a heavier molecular form of acetylcholinesterase sedimenting at 10.5S. The effect of other agents on the growth and differentiation of neuroblastoma cells is also presented.


In vitro studies on neural specificity.

In vitro studies on neural specificity. The migration of the retinal ganglionic axons through the optic nerve, over the tectal surface, and into the tectum to synapse correctly with brain neurons has been one of the most studied paradigms of neural specificity. Through the use of dissected tecta from the developing chick embryo, dissociated retinal cells can make the same choices in vitro that the ganglion cell termini make in vivo. That is, cells from the dorsal retina prefer to adhere to ventral tectum fragments and vice versa, in accord with the final map of the retina on the tectum. This assay has been used to analyze the biochemical components on the cell and tectal surfaces that might account for the recognition observed. Also, the assay has made it clear that virtually all the retinal cells can distinguish between dorsal and ventral tectal fragments, even though the cells of the retina that normally synapse with the tectum make up not more than 5% of the total population of the retina. One reason for this may be that, although only the retinal ganglion cells send their processes into the brain, these processes use all of the retina initially to get to the fundus of the retina, where the optic nerve begins. The vast majority of the retinal cells may possess surface recognition molecules not for finding the tectum but for serving as substrates for the few retinal cells whose axons must first leave the retina before finding the correct tectal locus.


Establishment of mouse neuroblastoma clone N18, rat glioma clone C6, and rat striated muscle clone L6 in serum-free, chemically defined medium. Mouse neuroblastoma clone N18, rat glioma clone C6, and rat striated muscle clone L6 were grown in a serum-free Dulbecco's modified Eagle's medium. Their doubling times were 48 hours, 40 hours, and 4 days, respectively. Morphologic features were similar to the original parent cell lines. Membrane components of N18 and C6 grown in serum-free medium were compared with the original parent cell lines. Defects of several membrane proteins were found with sodium dodecyl sulfate--polyacrylamide gel electrophoresis.


Surface membrane glycopeptides and cell differentiation. It is suggested that a correlation exists between the proportions of specific groups of glycopeptides that are found in the surface membranes and the ability of neuroblastoma clones to differentiate morphologically. The evidence for this is examined and a comparison is made with other properties of the differentiated cells.


Phenotypic modulation in mouse neuroblastoma cells: the case of acetylcholinesterase. Mouse neuroblastoma clone NIE-115 contains two species of acetylcholinesterase (AChE) which sediment at 4S and 11S. The former is predominant in growing cells (70%), whereas the 11S is more abundant in cells with neurites (55--65%). When cell replication is arrested by sodium butyrate, there is an increase in AChE activity but no modification in the proportion of 4S:11S species. Inhibition of protein synthesis by cycloheximide provokes a slow increase in the 11S form and a parallel decrease in 4S. Neurite retraction in presence of colchicine or excess serum does not lead to a concomitant reversion in the high 11S proportion. We postulate that the 11S AChE is formed by a conversion of the 4S and that the shift in the S-value ratio of the enzyme reflects changes in membrane remodeling during neurite extension.


Adenylate cyclase activity of normal and transformed fibroblasts in culture. The intracellular concentration of cyclic AMP regulates many cellular properties of normal rat, mouse, and human fibroblasts in culture. Thus it is important to elucidate how the activity of adenylate cyclase of fibroblasts might be altered and regulated by both intracellular and extracellular agents and events. In studies with several virally transformed fibroblast lines, as well as those transformed spontaneously or by 3-methylcholanthrene, a common feature of each type of transformation is a defective adenylate cyclase system. However, the means by which adenylate cyclase activity is altered differs with the cell system and the type of transformation. Here we concentrate on efforts to understand the regulatory properties of the adenylate cyclase from normal rat kidney fibroblasts. The modulation (increase or decrease) of the hormonal responsiveness of this enzyme may play an important role in its regulation. Of substantial interest is the isolation from serum of a high-molecular weight factor that selectively decreases the GTP and hormone-stimulated activities of adenylate cyclase. These findings are of value in our attempts at elucidation of the reasons for altered cyclase activity following virus transformation and during various stages of growth.


Recent developments with karyoplast regeneration and nuclear transplantation. Karyoplasts and cytoplasts prepared by cytochalasin-induced enucleation techniques were used in the development of systems for large-scale nuclear transplantation and the regeneration of karyoplasts to reform whole viable cells. Ten to 20% of a preparation of mouse L929 karyoplasts remained viable. These bodies, purified from whole cells, cytoplasts, and nonviable karyoplats, contained less than 10% of the cytoplasm in whole cells. By the technique of mass nuclear transplantation, up to 40% of a culture of cytoplasts could be renucleated by fusion of karyoplasts to a monolayer of enucleated cells. More than 5 x 10(5) renucleated or true cytoplasmic-nuclear hybrid cells could be prepared from a single cytoplast monolayer culture.


Cellular and molecular studies on globin gene expression. Globin gene expression has been studied with the use of a combination of cell and molecular biology techniques. With a somatic cell hybrid between mouse erythroleukemia (MEL) and human erythroid cells, human and mouse globin genes can be coexpressed in the same hybrid cell. Somatic cell hybridization between MEL cells and nonerythroid cells (e.g., human fibroblasts) results in hybrid cells that cannot be induced to produce hemoglobin of any type. Molecular hybridization of cellular DNA and RNA with globin cDNA indicates that the globin genes are present but that no globin mRNA is in the nonexpressing hybrid cell. This finding demonstrates that the loss of globin gene expression in the hybrid cell occurs at the level of transcription or mRNA processing. The nucleus of the nonerythroid cell is not necessary for the extinction of globin gene expression, since cybrids formed from MEL cells with enucleated nonerythroid cells also result in cells which cannot be induced to synthesize hemoglobin even after 40 or more generations. These data suggest that the cytoplasm of cells contains diffusible regulatory molecules which influence globin gene expression. In our attempt to develop an intracellular assay to test for regulatory molecules, we studied the procedure microinjection with red blood cell (RBC) ghosts. Rabbit globin mRNA (with hemin) was loaded into RBC's and then, by Sendai virus-mediated fusion, into Chinese hamster ovary (CHO) cells growing in culture; these microinjected CHO cells synthesized hemoglobin. Since this microinjection procedure appears to be effective in inserting both proteins and nucleic acids into intact cells, it should be possible to develop modifications for inserting aliquots of fractionated cytoplasm into growing cells. The procedure might then be useful as an assay for putative genetic regulatory molecules.


Use of an EK-2 vector for the cloning of DNA from higher organisms. A certified EK2 bacteriophage lambda vector, which is useful for cloning fragments of DNA from higher organisms, is described.


Amplification and characterization of eukaryotic structural genes. An approach to the study of eukaryotic structural genes which are differentially expressed during development is described. This approach involves the isolation and amplification of mRNA sequences by in vitro conversion of mRNA to double-stranded cDNA followed by molecular cloning in bacterial plasmids. This procedure provides highly specific hybridization probes that can be used to identify genes and their contiguous DNA sequences in genomic DNA, and to detect specific RNA transcripts during development. The nature of the method allows the isolation of individual mRNA sequences from a complex population of molecules at different stages of development.


Castration effects on tumor-specific immunity. Using 2 immunogenic (3-methylcholanthrene-induced fibrosarcomas in BALB/c x DBA/2 F1 (CD2F1) male mice, we observed initially that the rate of tumor growth might be enhanced by castration. For confirmation, tumor transplantation experiments with over 500 mice were done to compare tumor-specific transplantation immunity in castrate and in control male mice. Inbred mice bearing a 3-methylcholanthrene-induced fibrosarcoma transplant underwent surgical excision of the tumor; specific resistance to subsequent challenges with varying doses of that tumor cell line were compared in castrate and in noncastrate groups of mice. Although castration influenced the rate of tumor growth, it had no apparent effect on tumor-specific immunoresistance. Mechanisms of host-tumor immunorelationships are discussed as they might relate to endocrine therapy of prostate adenocarcinoma.


Monocyte chemotactic mechanisms in patients with hypernephroma. Peripheral blood monocyte chemotaxis was evaluated in 15 patients with hypernephroma, 10 of whom were evaluated preoperatively and postoperatively. Five patients had a significantly reduced response preoperatively that returned to normal levels after radical nephrectomy. Animal experiments confirm the presence of a low-molecular weight (6,000--10,000 daltons) inhibitor, which is heat labile and functions at the macrophage level. Preliminary experiments indicate inhibitors of macrophage chemotaxis can be extracted from renal cell carcinomas. The importance of these inhibitors in modulating immune surveillance is discussed.


Prostate carcinoma: mitogenic stimulation of lymphocytes with phytohemagglutinin. Lymphocyte transformation in response to mitogenic stimulation with PHA was studied in 46 patients with carcinoma of the prostate who have been followed for more than 3 years. This was related to clinical stage and histologic grade to form a prognostic index to forecast the natural history of this disease.


Factors that affect the results of microcytotoxicity testing in man. A great number of variables affect the pattern of test results obtained in studies of cell-mediated cytotoxicity of blood lymphocytes for tumor cells in vitro. The classic microcytotoxicity test measures both cell destruction and effects on cell growth. Microcytotoxicity tests in which isotopically prelabeled target cells are used measure primarily cell destruction. The method of preparation of effector cells from human blood can greatly influence results of such tests. Perhaps the most clear-cut example of this influence is the effect of Tris--NH4Cl treatment of effector cells to lyse contaminating red blood cells. This treatment inactivates the lytic function of certain effector cell types. In addition, a major consideration that must be kept in mind is the difference in patients under study by different groups of investigators.


Renal cell carcinoma: tumor membrane lymphocyte stimulation assay. A tumor plasma membrane LSA has been developed to measure cell-mediated immunity in patients with renal cell carcinoma. In this assay system, plasma membranes are adjusted to antigenic equivalence with the use of HLA as a plasma membrane marker. Fifteen patients were studied; 6 of 8 had a significant response to autologous tumor (P less than 0.05). Of 7 patients studied with allogeneic tumor, only 1 was responsive. Various parameters of the tumor membrane LSA are discussed, including dose response, kinetics, and method of data presentation.


In vitro generation of cytotoxic lymphocytes by tumor extracts of transitional cell carcinoma. Results of experiments in which in vitro generation of cytotoxic lymphocytes were used are presented. Problems of specificity and levels of cytotoxicity are also briefly discussed.


Correlation of primary tumor bed lymphocytic infiltration and peripheral lymphocyte cytotoxicity in patients with transitional cell carcinoma. Patients with transitional cell carcinoma were studied in an attempt to correlate lymphocyte infiltration into the primary tumor bed with peripheral lymphocyte cytotoxicity as measured by a chromium-51 release assay.


Evaluation of the relationships of prealbumin components in sera of patients with cancer. Serum proteins of cancer and control sera have been evaluated by the conventional Mancini method as well as by scanning densitometry measurements of components after separation by gradient PAGE. We established than in sera of cancer patients, prealbumin levels decrease and alpha 1-acid glycoprotein levels increase significantly. However, the Mancini method was not as sensitive as PAGE for quantitative evaluation of differences among sera of healthy adults, smokers, and patients with benign disease. We find that a more sensitive measure is the ratio between the two components (alpha 1-acid glycoprotein and prealbumin) and suggest that these values possibly may be useful as a CSI. In this study, the average CSI for healthy adults is 0.79, for heavy smokers, 1.24; for benign diseases, 1.67; and patients with various forms of cancer as follows: early lung, 4.17; late lung, 8.78; bladder, 8.92; colon, 13.77; breast, 2.43; and melanoma, 9.22.


Use of serum beta-chain human chorionic gonadotropin in the management of testis tumor. One-third of all testis tumors secrete HCG which can be detected in the serum by radioimmunoassay. beta-HCG levels can be used for diagnosis and follow-up of testis tumors. Seminomas with elevated serum beta-HCG levels probably have nonseminomatous metastases.


The carcinoembryonic antigen test in urologic cancer. The diagnostic value of the CEA test was evaluated in 2,029 blood and urine samples from 308 patients with urologic cancer, 13 with nonurologic cancer, 20 urologic patients with nonmalignant disease, and in 30 controls. The blood CEA test was positive in 50% of the patients with active urologic cancer and in 35% of those with inactive cancer. The urine CEA test was elevated only in patients with active bladder cancer, but most of these had concurrently infected urines. At the present time, the CEA test does not reflect the accuracy necessary for an acceptable diagnostic test for the detection of urologic cancer.


Transitional cell cancer: establishment and characterization of cell lines. Eleven long-term (in culture more than 1 yr) cell lines were established from surgical specimens of human TCC. Characterization studies performed on the individual cell lines showed that each 1) demonstrated an abnormal human karyotype, 2) grew in soft agar, 3) exhibited rapid growth and multilayering 4) was free from microbial and HeLa cell contamination, 5) produced tumors in cheek pouches of immunosuppressed Syrian golden hamsters, 6) contained ultrastructural features consistently found in epithelial cells in culture, and 7) could be grown to high cell densities in roller-bottle cultures.


Carcinoembryonic antigen as an adjunct to determination of clinical stage in prostate cancer. CEA level has been measured in a series of prostate cancer patients. Data are presented to show CEA correlation with clinical course and acid phosphatase level.


A radioimmunoassay for prostatic acid phosphatase. A solid phase radioimmunoassay for human prostatic acid phosphatase has demonstrated substantially greater biochemical sensitivity than a standard enzymatic method for which p-nitrophenylphosphate was used as substrate. Preliminary data indicate that the radioimmunochemical approach can precisely classify 43% stage I-II and 94% stage III-IV prostate cancers. In contrast, the standard enzymatic methods correctly classified only 9% stage I-II and 46% stage III-IV cancers. It is clinically apparent that a radioimmunochemical approach for the measurement of human prostatic phosphatase may have distinct potential in the clinical diagnosis of prostate cancer.


Biochemical and immunologic techniques for acid phosphatase measurement in the diagnosis of prostate cancer. A counter immunoelectrophoresis procedure for the measurement of acid phosphatase is described. Acid phosphatase was purified from human prostate. A monospecific antiserum to the purified enzyme was produced in rabbits. With the specific antiserum and a sensitive chemical staining technique, a counter immunoelectrophoresis assay was developed for detection of serum prostate acid phosphatase. The isoenzymes of serum prostate acid phosphatase were also studied by an isoelectric focusing technique.


Transitional cell carcinoma: circulating tumor antigen correlated with presence/absence of tumor (leukocyte migration assay). Sera from patients with TCC of the urinary bladder have been fractionated. We tested the fractions for activity in the leukocyte migration test, using peripheral blood leukocytes from TCC patients, other cancer patients, and healthy control donors as indicator cells. The results suggest that an antigen associated with TCC can be detected in the sera of patients bearing TCC but not in tumor-free individuals.


Skin tests with soluble tumor membrane antigens in patients with transitional cell cancers. Patients with transitional cell cancers and control patients with other forms of cancer were tested with cell membranes and soluble membrane antigens with the use of delayed hypersensitivity skin tests. These ongoing and parallel studies in which LMI tests are used have not been completed.


Characterization of prostate carcinoma lines in the Copenhagen rat. A comprehensive study of the R-3327 line of prostate adenocarcinoma of the Copenhagen rat was performed. This tumor, investigated in its histologic, endocrinologic, and immunologic aspects, was compared with a squamous cell prostate carcinoma derived from the R-3327. The two tumor lines differ in their rates of growth and in their androgen receptor contents, i.e., the adenocarcinoma is androgen dependent and grows slowly, whereas the squamous cell carcinoma has a rapid rate of growth and is androgen independent. A study of cell-mediated immune responses revealed that: 1) Nonspecific responses to mitogens in the blastogenic assay, as well as antibody-dependent cellular cytotoxicity, are enhanced in animals bearing tumors. 2) The R-3327 is immunogenic to its host as demonstrated by two parameters. 3) The antigens present in the squamous cell carcinoma are not recognized by lymphocytes of the animals bearing the adenocarcinoma, substantiating the specificity of the reaction. For these reasons, the system of R-3327 prostate adenocarcinoma provides a relevant model for the study of prostate cancer.