The diffusion of new hospital technologies in the United States. Increases in the real resources used in hospital care have been an important cause behind rising hospital costs in the United States. Many of these resources have taken the form of new hospital technologies, and this paper begins by reviewing the trends in adoption of new hospital technologies over the years 1950-1974. The resource requirements, costs , and to the extent possible the patient benefits, of two of these technologies are then discussed in more detail: intensive care, a widespread facility with many variations, has been a major contributor to hospital costs; radiotherapy has been characterized by a succession of competing technologies. Regulatory efforts such as certificiate-of-need reviews would be more effective if they viewed hospitals as flexible collections of such technologies-with the costs and patient benefits of each to be weighed separately-rather than primarily in terms of numbers of beds. A national center to collect information on the separate technological functions of hospitals and make it available to interested groups would make a useful contribution to hospital regulation.


The service economy and postindustrial society: a sociological critique. The growth of the service sector in advanced capitalist societies is the empirical evidence in support of the ideological approach to our social systems as "postindustrial." By examining the content of so-called "service activities" and by relating the observed trends to the model of uneven development in the U.S., this article tries to explain the transformation of the social structure of the U.S. on the basis of the contradications inherent in the processes of accumulation of capital, class struggle, and state intervention.


Breast-feeding: the role of multinational corporations in Latin America. The decline in birthrates in the developed countries of the world has forced multinational corporations engaged in the production of infant formula to seek out new markets in the developing countries, where burgeoning population rates potentially guarantee the long-term profitability of these corporations. This development, ostensibly benign and nutritionally advantageous to infants in developing countries, has serious public health consequences, due to the high relative cost of purchased formula and the paucity of hygienic facilities essential to the sterile preparation of bottle formula. This paper delineates in detail economic and contraceptive advantages of breast-feeding, and examines the role of health personnel and multinational advertising techniques which have catalyzed the decline in breast-feeding. In addition, the paper focuses on the question of cultural imperialism and current efforts to regulate the multinational firms through both United Nations groups and stock-holders' suits. Finally, some suggestions are made concerning ameliorative public policy approaches to the breast-feeding controversy.


Malpractice in the United Kingdom.

Malpractice in the United Kingdom. The end of the last century saw the development in the United Kingdom of medical protection and defense societies. Their history, objects, growth, and scope are outlined. Reference is made to leading cases of medical malpractice and to the relevant statutes. The relationship between the protection societies and the National Health Service authorities is dealt with and a comparison made of the relative situations in the United States and United Kingdom. Finally, recent trends are examined and views expressed about possible future developments in this field which will be the subject of recommendations by a Royal Commission on Civil Liability and Compensation for Personal Injury which is now sitting.


Nurses and trade unions in Britain.

Nurses and trade unions in Britain. The nature of the negotiating body for pay and conditions of nurses in Britain and some of the reasons for the body's past failure to negotiate a substantial pay increase for nurses are described in this article. A description is given of a series of events, including strike action by nurses during 1974, which led to a substantial pay raise. The pay award, however, represented only a partial success for the trade unions and the negotiating body. Methods of nurses' organization and the roles of their representative unions and professional body indicate some of the reasons for only a partial success. The lack of unity between the National Health Service trade unions and the reactionary role of the professional body were notable. However, many important political and organizational lessons were learned by nurses in this struggle. We are now facing savage cuts in National Health Service expenditure, leading to a reduction in the number of nurses employed. These lessons are clearly useful for the nurses and their organization with the labor movement in fighting these cuts in National Health Service expenditure.


Observations on potential contributions of health planning. The passage of the National Health Planning and Resources Development Act in the United States in 1974 is used to set the context for a new assessment of health planning as a change agent. In reviewing the record of health planning the most striking conclusion is that even its friends have been unable to establish that it has had any quantifiable impact. The authors suggest, however, that comprehensive health planning may have stimulated the belief that changes in medical care organization are crucial to improving the health care system. The authors next consider the role of health planning inferred from three widely espoused "models" of the health care system: professional, central planning, and market. Although market advocates generally deemphasize health planning as contrasted to those supporting a centrally planned system, none of the models is sufficiently developed to indicate specific roles and functions fro health planning. Basing their argument on goals for health care reform generally espoused by students of medical care organization, the authors assert that health planning agenices will be most effective if they are organizationally linked to general-purpose governments, encourage the formation of Health Maintenance Organizations, consciously involve themselves in health system reorganization, and design their policies so they can be effectively evaluated.


Toward the formulation of sociodental indicators. The bases for the construction of sociodental indicators is discussed in the paper, considering several available indexes of oral health status (dental caries, periodontal disease, malocclusion, oral hygiene, and other oral conditions) as well as measures of quality of services. Very little research exists relating any of the above measures to social indicators such as personal life-style or cultural and ecological factors. Such expansion would enable dental indicators to be useful for purposes of policy decisions. Combining any dental indicators or set of indicators with a potential global social health index is discussed in terms of potential problems obscuring dentistry's cost to society. Dentistry, in addition, is offered as a system in microcosm-one which can be useful for purposes of polishing methodology for the social health indicator movement.


Urticaria secondary to a copper intrauterine device. A 24-year-old woman developed in acute urticarial reaction secondary to a copper intrauterine contraceptive device. Allergy to copper was proven by scratch tests. The condition cleared with removal of the IUD.


Preoperative evaluation of the strabismus patient. The preoperative evaluation of the strabismus patient may require one or several examination sessions. Since general anesthesia is most often employed, the patient's general health, hematological and neurological status, and concurrent medications should be evaluated. The age of onset and whether the deviation was intermittent in character for some part of its course are important clues to fusion potential. The examination should emphasize close observation of the eyes when they are stationary and during rotations; it should be conducted so as to encourage, rather than frustrate, binocular cooperation. This requires test selection that takes into account both variations in alignment (at different test distances) and amblyopia. Sensory evaluation can involve many tests, but specifically for surgical planning it can be brief and apply principal to justification of surgery for small-angle, nonsymptomatic deviations. The phenomenon of intractable diplopia is unusual and probably should not be considered a strong deterrent to surgical correction of a long-standing and cosmetically unpleasant misalignment.


Surgical weakening of the inferior oblique. 1. The inferior oblique may be weakened effectively by recession, disinsertion, or myectomy, disrupting the muscle continuity between Lockwood's ligament and the muscle's insertion. 2. A successful unilateral inferior oblique weakening produces 20 prism diopters less hypertropia or more hypotropia in the field of action and has less effect away from the muscle's field of action; a bilateral weakening of the inferior oblique produces 20 prism diopters less exotropia or more esotropia in upgaze. 3. Complications of inferior oblique weakening are persistent overaction, operation on the wrong muscle, and the adherence syndrome. 4. The adherence syndrome is not related to the myectomy procedure specifically but is probably related to (or caused by) fat rupture with hemorrhage, which may accompany any type of inferior oblique weakening. 5. The adherence syndrome can be avoided by careful surgical technique.


Superior oblique palsy: diagnosis and classification. A review of 130 cases of superior oblique palsy suggests that simple office procedures are sufficient to establish the diagnosis. Prism and cover test measurements with alternate eyes fixing in the nine diagnostic positions of gaze proved to be the greatest help in classifying these cases with regard to surgical planning.


Diagnosis and surgical options in superior oblique surgery. The surgical treatment of superior oblique paresis is most gratifying to the patient and to the surgeon. Whereas the large range of vertical fusion may make it difficult to bring out the total deviation, this same fusion is the surgeon's best ally postoperatively. In addition, while there are multiple choice as to which muscle or muscles to operate upon, the great advantages of the tucking operation on the superior oblique are (1) it practically always straightens the head, (2) it greatly reduces the hyperdeviation, and (3) it has no effect on the width of the lid fissures.


Superior oblique tenotomy: indications and complications. Two series of patients who had superior oblique tenotomies either alone or in combination with horizontal muscle surgery were evaluated, and the results and complications were analyzed. The effect of bilateral superior oblique tenotomies in downgaze correlated well with the previously reported series. In these two series, the effects in primary and upgaze also were evaluated. An average effect of 12 prism diopters eso-shift was found to occur in primary position, while no eso- or exo-shift was found in upgaze. Operative indications and contradictions, when followed carefully, make predictable the effectiveness of the operation in collapsing an A pattern. The reported unequal effects on superior oblique action were not observed in any of the patients studied. In no instance in these series was there a worsening of preexisting hypertropia or development of a new one. Transient sheathlike Brown's syndrome occurred postoperatively in some patients in the early series, but this problem was reduced by the instillation of steroid suspensions. V patterns developed postoperatively in 20 percent of the cases in the first series and in 9 percent in the second series. There was approximately a one-in-three chance that the inferior oblique muscle would become slightly to moderately overactive postoperatively. The incidence of this complication will be reduced significantly or eliminated by careful case selection to choose only those patients who have slightly underactive inferior oblique muscles preoperatively.


The role of the fascia in muscle surgery. Mishandling of the fascial tissues during surgery on the extraocular muscles accounts for unsatisfactory alignment of the eye in the primary position; restricted movement of the globe; scarred, injected, heaped conjunctiva; and changes in the normal eyelid level. These hardships may be impossible to overcome by performing secondary surgery. The most serious and frequently recurring abuse of the fascia during muscle surgery is the needless penetration of the Tenon's capsule beyond 10 mm. from the limbus, which results in prolapse of the fat cushion into the spaces that are normally between the movable fascial planes and sclera. The prolapsed fat infiltrates into the sclera and muscle capsules and undergoes fibrofatty proliferation; with time, cicatrization of these scarred tissues pulls the eye to a deviated position, restricting its normal movement. Also, the fat cushion migrates forward under the conjunctiva toward the limbus, causing the unsightly appearance of injected yellowish fleshy tissue near the limbus. A patient's ocular motility can be ruined for life and his appearance deformed because of a surgeon's disrespect for the important role Tenon's capsule plays in the motility of the globe.


Surgery of dissociated vertical divergent strabismus. In those cases where definite bilateral inferior oblique overaciton coexists and predominates, the initial procedure should be a maximum weakening operation of both inferior oblique muscles. For the remainder, surgical treatment should be reserved for the unsightly predominantly unilateral cases who do not wish to wait for the possible beneficial effects of age. With surgery on the vertical rectus muscles, particularly the inferior rectus, improvement of the deviation by two-thirds to three-quarters should be the aim of treatment.


Role of Tenon's capsule in postoperative restrictions. Management of all restricted strabismus cases is difficult and frequently will require two or more procedures. The prevention of these restrictions at the time of primary surgery is far more important. All tissue should be handled as gently as possible, with smooth or fine-tip forceps, avoiding large fixation devices. Sharp dissection is essential - tissue should not be torn but cut carefully and sharply with scissors under direct visualization. The conjunctival and Tenon's capsule incisions should not be made directly over the muscle; this will result in scarring of the muscle to Tenon's capsule or of Tenon's capsule to the muscle insertion stump, promoting restriction. The muscle should be isolated under direct visualization or inspected carefully after isolation, with great care taken not to incorporate any unnecessary Tenon's capsule in the muscle and to avoid the inferior oblique msucle on lateral rectus muscle surgery. A blind, deep sweep is completely contraindicated. The intermuscular membranes, check ligaments, and muscle foot plates must be completely severed, under direct visualization, well beyond the resection site in the course of a resection,and 5 to 8 mm. behind the insertion in the course of a recession. Complete hemostasis during the procedure and at its completion is essential; 2.5 percent phenylephrine and minimal cautery will help achieve this. Since cautery will result in a scar of Tenon's capsule to that area of the sclera, it should be used as little as possible. Phenylephrine 2.5 percent has been used safely with halothane anesthesia in many patients without significant arrhythmias. The secondary treatment of restriction is very difficult, whereas their prevention requires only good surgical technique and careful attention to detail.


Special considerations and techniques in strabismus surgery. Strabismus surgery continues to present a challenge to the ophthalmologist who evaluates results critically. Precise diagnoses and expert surgical judgment never will be supplanted by tools and techniques. Nevertheless, by remaining aware of newer surgical procedures and improved instrumentation, the strabismus surgeon will continue to increase the probability of achieving desirable surgical results.


Surgery for congenital esotropia.

Surgery for congenital esotropia. The diagnosis of esotropia in an infant is not generally a difficult one for a skilled ophthalmologist to confirm. Uncovering all the subtleties of the disease presents the greater challenge and taxes the skills involved in examining infants. The numerous treatment regimens available leave the surgeon who does this operation only occasionally in doubt as to when it is best to operate and by what method. The safest guidelines for the ophthalmologist are (1) to be sure of his diagnosis and all of its ramifications and (2) to operate on these patients by the method he feels will best totally correct both the horizontal and vertical defects in one operation. It is also known, however, that not all of these infants will be corrected by this single procedure and that additional surgery, both early and delayed, will be necessary. Similarly, amblyopia and accommodative esotropias will need to be treated in a significant segment of these patients. The stability of the final result depends upon the success of the treatment in all parameters. The results obtained are far from ideal. Some patients achieve the optimum with good and equal acuity in each eye, straight eyes, peripheral fusion, fusional amplitudes, and reduced stereopsis. Other patients will have reduced visual acuity secondary to amblyopia; deficient or no binocular cooperation; and larger eso-angles or even consecutive exo-angles - both of which will require further surgery. Congenital esotropia is a fascinating entity that requires more research before its etiology and pathogenesis will finally be understood. Only then will therapy produce a satisfactory and lasting result.


Smooth muscle regeneration in swine ureters: a light and electron microscopic study. Intubated ureterotomies were performed on 16 juvenile pigs. The animals were killed at intervals of from 2 to 91 days after operation and tissue sections examined with light and electron microscopes. Ureteral wall replacement occurred within the 3 months after operation and included all tissue layers. Epithelialization of the wound surfaces occurred in the 1st week and was accompanied by a connective proliferation. Smooth muscle bridged the defect within 5 weeks and the margins of the muscularis were thickened and cellular in early stages; cells were elongated and extended towards the defect from the muscular remnant. Primitive smooth muscle cells were present at defect margins. This evidence indicates that smooth muscle was replaced by hyperplasia.


Repair of the injured renal papillary tissue in rats. Renal papillary necrosis was produced in rats by administration of 2-bromoethylamine hydrobromide. The rats drank either 0.9 per cent sodium chloride solution or distilled water, and structural restoration of renal papilla and effect on urine-concentrating ability were examined. All rats drinking distilled water had no evidence of structural restoration of the injured papilla and no significant improvement of urine-concentrating ability. When rats started drinking 0.9 per cent sodium chloride solution shortly after indution of the necrosis, enhanced cellular reconstitution of the necrosed papillary tissue and partial restoration of urine-concentrating ability occurred in a significant number of rats. The most successful structural reparation was the collecting tubules, followed by the thin loops of Henle and the capillaries. Repopulation of the interstitial cells was the least successful. In conclusion, repair of necrosed renal papilla was enhanced when rats drank 0.9 per cent sodium chloride solution starting shortly after injection of renal papilla-necrotizing dose of 2-bromoethylamine hydrobromide.


The epitaxially induced crystal growth of calcium oxalate by crystalline uric acid. Seed crystals of anhydrous uric acid were shown to epitaxially induce the crystal growth of calcium oxalate monohydrate from its metastable supersaturated solution. Scanning electron microscopic examination of crystalline material isolated from the kinetic experiments supports the chemical evidence that calcium oxalate had been nucleated by uric acid. In spite of close lattice similarities between some of the faces of the two phases, the induction period required for heterogeneous nucleation by the seed material was about half of that required for precipitation in the absence of potential nucleating agents. This has created some doubt as to whether crystalline uric acid has an influence on the formation of calcium oxalate lithiasis.


Comparative effects of anticalculus management in the rat. Utilizing a low pyridoxine, ethylene glycol-fed rat model, various modes of anticalculus therapy were compared. Prophylactic papillary and parenchymal oxalate stone therapy is most effective with magnesium oxide. Hydrochlorothiazide is next in order as the therapy of choice in this group. A high calcium diet seems to negate the effect of magnesium oxide. Methylene blue, magnesium oxide, and even sodium phosphate, are not as effective as hydrochlorothiazide in the combined prevention and treatment of preformed parencymal calcium oxalate stones. No agent seems to have a profound effect on preformed apical papillary calculi.


Effect of tetracycline on calcium oxalate calculi: in vivo and in vitro studies. Inadvertent use of tetracycline hydrochloride while producing calcium oxalate stones in the rat resulted in a reduction of papillary concretions by one-half. Parenchymal calcifications in the continuing long term, preformed stone group were reduced to levels comparable to that of other useful agents. In vitro studies corroborated these findings at physiologic urinary pH ranges. The rationale and comparisons to other anticalculus drugs are discussed.


The absorption metabolism, and excretion of Estracyt (NSC 89199) in patients with prostatic cancer. Estraumustine phosphate (estradiol, 3-N-[bis(2-chloroethyl)-] carbamate-17beta-dihydrogenphosphate (Estracyt) labeled with 3H in the estradiol moiety and 14C in the carbamate moiety was synthesized, and its absorption, metabolism, and excretion were studied after oral administration to three patients with prostatic carcinoma. One of the patients was also given the same dose by intravenous injection. In addition to monitoring isotope levels in peripheral blood, urine, and feces, samples of portal vein blood were obtained through a catheter in the umbilical vein. Analyses of portal blood samples revealed that most of the estramustine phosphate was dephosphorylated to estramustine during absorption. Estramustine was found to be the major metabolite in peripheral blood after oral as well as after intravenous administration. The urinary excretion data appeared to warrent the conclusion that most of the carbamate ester of estraumustine is hydrolyzed before it is excreted. In the patient given estramustine phosphate by both routes the absorption of the compound when given by mouth was found to be approximately 75 per cent.


Substrate metabolism in the isolated perfused dog kidney. The relationships between substrate utilization and renal work were investigated in an isolated, perfused dog kidney preparation: substrate utilization included free-fatty acid (FFA), glucose, and oxygen, whereas work was assessed by measuring TNa+. The magnitude of TNa+ was varied by altering glomerular filtration rate as a consequence of changing renal blood flow. As the rate of renal blood flow was reduced, both Qo2, and QFFA decreased while Qglucose increased. The reduction in QFFA and Qo2 paralleled the decrease in TNa+. An inverse relationship was noted between QFFA and Qglucose in all experiments. The results suggest that FFA and perhaps glucose may serve as a significant substrate for providing energy for TNA+ in the kidney. Further, since QFFA and Qglucose are inversely related to each other, glucose may substitute for FFA as an energy source at times when QFFA is reduced.


An experimental study of the calcification and absorption of polyglycolic acid and catgut sutures within the urinary tract. This experimental study has shown that there is no difference in calcification on catgut and polyglycolic acid suture loops within the sterile and infected rat bladder. The presence of an experimentally induced infection reduces the time both catgut and polyglycolic acid sutures remain attached to the bladder wall. Polyglycolic acid sutures can nevertheless be recommended for routine use in the urinary tract whenever an absorbable suture is required in view of its greater strength, reliability and remarkable lack of inflammatory tissue reaction.


Role of neurogenic factors in canine urethral wall tension and urinary continence. A plastic disc, secured submucosally, was inserted in the canine bladder to permit filling of the bladder neck region while bypassing the accommodating ability of the detrusor. The bladder neck region was distended with radiopaque solution and the onset of flow in the urethra and the width of the urethra at steady state flow conditions were monitored by fluoroscopy. Perfusion pressure was also recorded. Bilateral section of the hypogastric nerves produced a substantial reduction in the time to initial flow and an increase in the width of the urethra with a decrease in pressure at steady state flow. Bilateral section of the pelvic nerves or administration of atropine reduced urethral wall tension at steady state flow but did not significantly influence the onset of initial flow. Gallamine treatment had no significant effect either on wall tension or on the onset of initial flow. It was estimated that the sympathetic nervous system contributed about 30 per cent to the urethral wall tension at steady state flow conditions whereas the parasympathetic nervous system contributed about 10 per cent. The remaining 60 per cent was attributed to the elastic properties of the urethra. In the maintenance of urinary continence, the sympathetic nervous system was estimated to contribute almost 60 per cent based on our experimental system. No other neurogenic influence was found to contribute significantly. Elastic and mechanical (anatomic) factors were probably responsible for the remaining 40 per cent. It was concluded that the sympathetic nervous system has a significant role in the maintenance of urinary continence.


Comparison of rapid versus slow decompression of the distended urinary bladder. Experiments are described showing that urinary retention with its associated increased intravesical pressure and overstretching of the bladder wall can lead to loss of capillary and tissue integrity. Our findings indicate that the severity of damage to the bladder wall prior to catheterization determines the degree of hemorrhage after bladder decompression and not the rate of emptying.


Total replacement of part of the canine urethra with lyophilized vein homografts. Three centimeters of the urethra were replaced by lyophilized homograft veins in nine male dogs. The dogs were followed for up to 6 months with periodic urethrograms. The grafted lyophilized veins served as a guide for the ingrowing transitional cell epithelium. The graft caused very little tissue reaction and hardly any formation of fibrous tissue. The urethra was covered by transitional cell epithelium after 4 to 8 weeks, and eight of the nine dogs showed no signs of obstruction during the follow-up period.


Incorporation of 3H-thymidine and 14C-amino acids into the ventral prostate after in vivo treatment with estradiol-3N-BIS(2-chloroethyl)carbamate-17 beta-phosphate (Estracyt) and its estrogen and cytostatic parts. The in vivo effect of Estracyt (a nitrogen mustard linked to estradiol-17 beta-phosphate) on testosterone-stimulated incorporation of 3H-thymidine and 14C-amino acids into the ventral prostate of castrated rats has been investigated. A dose of 100 mg of Estracyt per kg of body weight injected intravenously daily for 3 days significantly reduced isotope incorporation. Equivalent amounts of the two main components of Estracyt, the nitrogen mustard and estradiol-17 beta-phosphate, were not able to mimic this Estracyt effect. Possible implications of this difference and the intracellular mechanism of action of Estracyt are discussed.


The influence of hyperbaric oxygenation on rat testes. Using Wistar stain male rats, it has been shown that 2 atmospheric pressures associated with oxygen saturation, together or singly for 90 minutes daily for successive 46 days, resulted in no significant change on the ratio of testes to body weight. No significant change was observed in the incorporation of 3H-thymidine into testes and pituitary glands between the pressure-treated groups and the control group.


Osteoporosis and fractures of the neck of the femur: some epidemiologic considerations. The prevalence of osteoporosis in Jerusalem during the years 1967-71 was studied in a general population sample and in patients with fractures of the neck of the femur. Osteoporosis was determined by qualitative examination of lateral X-ray films of the lumbar spine. The prevalence of definite osteoporosis in a random population sample was 8.6%, and was higher in females (14.5%) than in males (2.9%). The prevalence of definite osteoporosis among 389 patients with fractures of the neck of the femur was 32.2%--37.8% in females and 17.4% in males. The results of this survey support the conclusion that osteoporosis plays an important role in the pathogenesis of fractures of the neck of the femur.


Menopausal bone loss and estrogen replacement. Throughout adult life the bone mineral mass of the radius is greater in males than in females. In males, it decreases after 60 years of age, while in females, it decreases earlier, at approximately 50 years, and the loss is greater. At the average age of 67 years, one half of the normal white female population has less than the normal amount of bone in the radius. Premenopausal women over the age of 50 do not show any decline of bone mineral mass, while in postmenopausal women, regardless of age, there is a loss of bone mass related to the number of years after menopause. Castrated women have significantly less bone mass than premenopausal women of the same average age. No decrease in cortical thickness of the radius was found in oophorectomized women treated with estrogens after castration. In a long-term, follow-up study, untreated postmenopausal women (after a natural or an artifical menopause) showed a significant loss of bone mass, while estrogen-treated, postmenopausal women showed no such loss. Estrogen treatment thus appears to prevent postmenopausal bone loss.


Prevention of bone loss following oophorectomy in premenopausal women: a retrospective assessment of the effects of oophorectomy and a prospective controlled trial of the effects of mestranol therapy. Prospective studies of bone mass in women following oophorectomy for benign conditions were done by the double-blind technique. Skeletal response to treatment was measured by photon absorption densitometry. Untreated patients were found to lose bone mass rapidly during the first two years after oophorectomy. When estrogen replacement was started within two months of oophorectomy, it was found to be effective in preventing subsequent bone tissue loss. Three years following oophorectomy, untreated women who had already lost bone tissue, and who were then started on estrogen replacement, showed a highly significant increase in their bone mass. The women in whom this treatment was delayed for six years did not respond. No untoward effects were noted in these women, perhaps, in part, because they had undergone hysterectomy. Long-term effects of this treatment are now being evaluated.


Role of hormonal factors in the pathogenesis of postmenopausal osteoporosis. In 47 women with postmenopausal osteoporosis, pretreatment studies by microradiography, radioimmunoassay and other methods showed increased bone resorption, normal bone formation, and decreased serum immunoreactive parathyroid hormone (PTH). In patients treated with a physiologic replacement dose of estrogen, bone resorption decreased to normal and PTH increased after short-term therapy; bone formation decreased to very low levels after long-term therapy. These data indicate that, in most patients, both an intrinsic abnormality of bone cell function and a disruption of the normal regulation of bone turnover by PTH and sex hormones, as a result of the menopause, are important in the pathogenesis of osteoporosis.


Relation to osteoporosis of age- and hormone-induced changes in the metabolism of collagen and bone. Experimental evidence is presented which suggests that age-induced changes in the collagenous matrix, the main constituent of the organic portion of bones, are at least partially responsible for age-induced physiological osteoporotic changes in the skeleton. In particular, there seems to be a labile fraction of recently synthesized collagen in bones, which loses its metabolic activity rapidly with advancing age. Experimental and clinical hormonal disorders and disturbances in calcium metabolism also cause changes in skeletal metabolism; these changes seem to be largely mediated through changes in the collagenous matrix. In experimental hyperthyroidism and hyperparathyroidism, the rate of degradation of the collagenous matrix appears to act as a moderator or "final messenger" in hormone-induced bone resorption. In conditions with altered calcium metabolism, such as malabsorption associated with hypocalcemia, altered bone metabolism may be due to osteomalacia or hypocalcemia-induced hyperparathyroidism. An increase in the rate of bone destruction in relation to the rate of bone formation is probably also the cause of postmenopausal osteoporosis. At present there is no optimal form of hormonal treatment for age-induced or post menopausal osteoporosis. Estrogen replacement therapy may be the best available treatment for postmenopausal osteoporosis, but slowing down the already low rate of bone catabolism in elderly subjects by estrogen or other therapeutic means requires long periods of treatment before pronounced increases in the total mass of bones take place and prophylactic administration of estrogen may produce better results.


Effect of conventional and newer forms of treatment on calcium absorption in osteoporosis. Studies carried out in this Research Unit and by others have shown that patients with osteoporosis absorb less calcium from a high calcium intake than do patients without osteoporosis. To determine whether various treatment modalities would increase the intestinal absorption of calcium, the effects of conventional and new methods were investigated. Tracer doses of 47Ca were given orally to determine the absorption of calcium. Calcium balances were also determined. Estrogen decreased urinary calcium and testosterone increased the nitrogen balance. Vitamin D and strontium increased the urinary calcium but did not increase calcium absorption. Of the newer compounds, fluoride did not affect calcium absorption but decreased urinary calcium. Phosphate had a similar effect. Infusions containing up to 15 mg calcium/kg, given for 6 or 10 days, increased the retention of calcium but did not increase its absorption.


Rehabilitation in postmenopausal osteoporosis. Rehabilitation and nursing care of patients with postmenopausal osteoporosis require a comprehensive team effort. The primary purpose of rehabilitation is to restore these elderly women to an optimal level of well-being and activity and return them to home and community life as soon as possible. Medical management, rehabilitative nursing and physical therapy are interdependent. Emphasis is placed on estrogen replacement to prevent further fractures and height loss, individualization of care, early ambulation, improving patient confidence, physiotherapy procedures, orthopedic supports, optimal nutrition, safety precautions, work simplification, occupational therapy and the need for appropriate social contacts. The economic, physical and emotional consequences of deforming postmenopausal osteoporosis are enormous. Current research indicates that this disease is preventible. An effective program of prophylaxis is needed now; otherwise many more women will need increasing amounts of costly rehabilitation.


Unsaturated fatty acids in adipose tissue of Israeli Jews. The ratio of polyunsaturated to saturated fatty acids in the adipose tissue of Israelis was found to be 0.88 for Ashkenazic Jews and 1.13 for non-Ashkenazic Jews, the highest reported for any population on a free-choice diet. Despite the unusually high ratios, indicating a high consumption of polyunsaturated fat, there is nevertheless, a relatively high incidence of ischemic heart disease among Israelis.


Morphological and cytochemical investigation of human mesothelial cells from pleural and peritoneal effusions: a light and electron microscopy study. We investigated the morphological and cytochemical properties of mesothelial cells obtained from pleural and peritoneal effusions of patients with various diseases. The morphological features were studied by light and electron microscopy and by cytochemical methods. The different kinds of contact between mesothelial cells and their interaction with lymphocytes were observed and the transition of mesothelial cells to macrophages and/or fibroblasts was demonstrated.


Apparent total alpha1-antitrypsin deficiency: report of a case. A 29-year-old female, with chronic renal failure and chronic bilateral emphysema, was admitted with severe uremia and septicemia secondary to multiple abscesses in the right kidney. Her condition improved after right nephrectomy. Pulmonary function studies showed marked obstructive and restrictive lung disease consistent witht the diagnosis of primary emphysema. On biochemical and histological examination, the liver was found to be normal. Alpha1-antitrypsin could not be demonstrated in the patient's serum at normal pH by any of the known techniques, but protein molecules with alpha1-antitrypsin antigencity were found at pH 4.8; this suggests a pH-dependent structural difference in alpha1-antitrypsin protein. Starch gel electrophoresis gave a multibanding pattern not previously described. A new form of apparent total alpha-1-antitrypsin deficiency is postulated.


Tetrazolium salts: a consumer's guide.

Tetrazolium salts: a consumer's guide. The purities of seven tetrazolium salts, obtained from various commercial sources, have been assessed by thin layer chromatography, relative extinction coefficients, and melting points. MTT and INT were largely homogeneous on thin layer chromatography, although significant variations occurred in the melting point behaviour. All the samples of TT examined were contaminated to a small extent with non-tetrazolium u.v.-absorbing material. TNBT and NBT were contaminated with small amounts of mono-tetrazolium salts, although one sample of each was heavily contaminated with another di-tetrazolium compound. Four samples of TNBT contained high melting point contaminants. BT was also contaminated with mono-tetrazolium salts, and some samples also contained di-tetrazolium salt contaminants. NT was the most heavily contaminated of all, most samples containing no less than five separate tetrazolium compounds. Prices varied widely, and in general were not related to purity. Some catalogue entries were very easy to find; others were more difficult. Few specifications were given; of these, most were arbitrary (for example, pure, grade I, and ... probably the finest INT offered anywhere.


The composition of Erythrosins, Fluorescein, Phloxine and Rose Bengal: a study using thin-layer chromatography and solvent extraction. Commercial samples of Erythrosin B (CI 45430), Erythrosin Y (CI 45425), Fluorescein (CI 45350), Phloxine (CI 45410) and Rose Bengal (CI 45440) have been analysed by thin-layer chromatography. The Erythrosins were found to be mixtures consisting in the main of 4'-iodofluorescein, 4', 5'-di-iodofluorescein, 2', 4', 5'-tri-iodofluorescein and 2',4',5',7'-tetraiodofluorescein, in some instances together with 2',4',5'-tri-iodo-4,5,6,7-tetrachlorofluorescein and 2',4',5',7'-tetraiodo- 4,5,6,7-tetrachlorofluorescein. Samples of Fluorescein were mixtures of the nominal dye usually with traces of several unidentified, fluorescent components. Those of Phloxine consisted mainly of mixtures of 4'-bromo-4,5,6,7-tetrachlorofluorescein, 4',5'-dibromo-4,5,6,7'-tetrachlorofluorescein, 2',4',5'-tribromo-4,5,6,7-tetrachlorofluorescein and 2',4',5',7'-tetrabromo-4,5,6,7-tetrachlorofluorescein, often with 4,5,6,7-tetrachlorofluorescein. Samples of Rose Bengal were mixtures of 4'-iodo-4,5,6,7-tetrachlorofluorescein,4',5'-di-iodo-4,5,6,7-tetrachlorofluorescein, 2',4',5'-tri-iodo-4,5,6,7-tetrachlorofluorescein and 2',4',5',7'-tetraiodo-4,5,6,7-tetrachlorofluorescein together with some unidentified components. Most of the commercial dye samples gave an insoluble residue when extracted with methanol. This residue was usually inorganic carbonate or halide. Some possible practical consequences of the various impurities are discussed.


The quantification of formazans in tissue sections by microdensitometry. II. The use of BPST, a new tetrazolium salt. This article describes the use of a microdensitometer for the measurement of BPST formazan in tissue sections. BPST is a new tetrazolium salt, 2-(2-benzothiazolyl)-3-(4-phthalhydrazidyl)-5-styryl-tetrazolium chloride, which produces a single, well-defined formazan, and is thus easily quantified. The formazan gives an excellent localization, since BPST was originally designed for ultrastructural work. Activities are expressed in absolute units as n moles hydrogen/mm3, and are thus directly comparable with standard biochemical data.


The quantification of ormazans in tissue sections by microdensitometry. III. The effect of objective power and scanning spot size. In this article, it is reported that the formazans derived from neotetrazolium chloride (NT) and 2-(2-benzothiazolyl)-3-(4-phthalhydrazidyl)-5-styryl-tetrazolium chloride (BPST) can be measured by microdensitometry of tissue sections with a wide range of scanning spot sizes, without any significant effect on the recorded extinctions. The spot sizes tested ranged from 0.20 to 16 mum. Thus when large fields have to be measured, it is possible to use low-power objectives and still obtain valid results.


Acid phosphatase and peroxidase in "resting" acinar cells of the major salivary glands of cats and their possible movement into secretory granules. After fixation of perarterial perfusion using an aldehyde mixture, salivary tissues were prepared for ultrastructural cytochemistry of acid phosphatase or peroxidase. Great variations in the distributions of the reaction product occurred, often within the same cell. Acid phosphatase staining occurred not only in lysosomes and sometimes in a GERL system, but a diffuse cytoplasmic component was also found in submandibular central acinar cells and to a lesser extent in parotid acini and variable staining occurred in the secretory granules of these cells. Peroxidase was variably associated with rough endoplasmic reticulum in submandibular demilunar cells, parotid acini, and more strongly in some sublingual cells. The secretory granules of the latter were darkly stained, but in parotid granules there was variable staining and least staining occurred in the granules of submandibular demilunes. These results are thought to indicate that not all enzymes present in secretory granules have reached there by an elective secretory process. Sometimes they appear to have entered the granules haphazardly, possibly having been enzymes associated with intracellular cisternal channels for transport or metabolism of other secretory substances and ultimately to have passed into the cisternal channels by chance or as part of a natural removal of redundant material.


Uridine diphosphoglucose dehydrogenase in the intervertebral disc. Intervertebral discs of an old sheep and a young pig were examined for the presence of cells containing the enzyme uridine diphosphoglucose dehydrogenase. In the sheep, the inner anulus had a higher proportion of active cells than the outer anulus; in the pig, there was no difference. From a consideration of cell numbers, it is suggested that there is an accumulation of glycosaminoglycans in the centre of the disc rather than an increased production rate. Notochordal cells in the pig disc contain uridine diphosphoglucose dehydrogenase and are capable of producing glycosaminoglycans.


[Electro-cochleographic findings in posterior fossa tumours (author's transl)]. The different audiometric and electrocochleographic findings in two patients with acoustic neuromas are presented. Significantly different thresholds from the two methods of testing are explained on the basis of the ganglion cells, peripheral axons and hair cells remaining unaffected for some time whilst the central axons are damaged by tumour pressure. Differences of recruitment measured conventionally by ABLB audiometry or by electrocochleography are explained on a neurogenic basis. In cases of acoustic nerve fibre damage, single fibre tuning is altered to a raised threshold and a reduced frequency specificity.


[How important are lower tunnels in the cottle septoplasty? (author's transl)]. The Cottle method is the best for correction of anterior nasal septal deformities. Establishment of the lower tunnel however seems unadvisable because of the common damage to the nasopalatine nerve passing through the incisive foramen with occasional permanent sequelae. The procedure also is time-consuming. As a rule spurs or deviations can be dealt with after establishing upper tunnels without perforation of the mucoperichondrial or mucoperiosteal flaps. The method is described in details.


[Animal experiments on the influence of sarbitol on the damaged internal ear (author's transl)]. The influence of sorbitol was studied by recording the microphonic potentials (MP) of 10 guinea-pigs whose internal ears had been noise-damaged immediately before. Neither the rise of MP 10 sec after the start of noise exposure nor the total rise of MP showed any changes after the intraperitoneal injection of 2 ml of a 40% sorbitol solution.


[Radiogenic sarcoma of the thyroid (author's transl)]. A sarcoma in the region of the thyroid, observed 30 years after radiotherapy in a patient with syringomyelia is presented. Even after low dose radiation in childhood and adolescence tumours of the thyroid frequently can be diagnosed. They rarely occur in adults, because apparently of a long latent period. This case is thought not to be a specific thyroid tumour.


[Applied phoniatry. I. Retarded language and speech development (author's transl)]. To better understand the assessment of difficulties in verbal communication it is considered to explain language and speech development in relation to motor function. The aetiology, diagnosis, prognosis and treatment of language disorders and their common characteristics are discussed. The otolaryngologist must expect an increase in the number of such cases and should initiate prompt and adequate treatment.


[Panel discussion: problems of the specialist's duty to inform the patient (author's transl)]. The obligation of the physician to inform the patient--which he has to prove in case of a suit--is based on the patient's right of self-determination. This self-decision information was subject of the panel discussion. Not discussed in detail were the information concerning diagnosis and prognosis, and the instruction of the patient regarding his conduct postoperatively and during medical treatment. Not considered was the so-called malpractice and negligence respectively. Medical liability suits are increasing for various reasons and are frequently directed at a failure to inform the patient becuase the patient is often unable to prove a negligence of the physician ("surrogate liability"). The dimension of the duty of disclosure (complete information--no information at all) is discussed in general and with special regard to the Ear-Nose- and Throat field. Special questions are answered regarding otoplasty, middle ear surgery, transplantations, extension of laryngeal surgery without prior informed consent, paranasal sinus and rhino-basis operations as well as surgery in minors and foreigners. In cases of non-vital indication and particularly in plastic-cosmetic procedures and outsider methods the requirements for informed consent are particularly important. The so-called rate of complications has perhaps a relative, but never an absolute meaning, and even this only in connection with the other circumstances of an individual case. The evidence of an adequate informed consent which must be included in a physician--patient-dialogue is most convincing by means of a written consent and the additional signature of a witness, perhaps also of the patient. The value, the problems and even the risks of an information based only on forms or brochures is discussed in detail. The panel discussion from the four points of view of the organizers should be no means confuse the otorhinolaryngologist. It should inform him about the medicolegal aspects of his activity and protect him from avoidable burdens.


[Transtympanic neurectomy of the vestibular and cochlear nerves (author's transl)]. Tympanotomy and a transvestibular approach to the fundus of the internal auditory meatus is described. The surgical technique is modified from that proposed by Alvarez de Cozar and Antoli-Candela in 1970. The surgical anatomy of the operative sites is detailed. The surgical procedure is found indicated in patients in whom preservation of hearing is not considered and vestibular neurectomy required. The described technique provides a simple but efficient and safe alternative to the translabyrinthine approach to the internal auditory meatus for division of the vestibular and cochlear nerves.


[The lombard reflex as a test of vocal function (author's transl)]. Any impairment of audio-phonatory control by background noise is followed by an increase in both the intensity and pitch of the speaking voice (Lombard reflex, 1911), thus increasing vocal strain. As a consequence, it might be anticipated that persons reacting to noise with marked changes in voice might be more liable to develop dysphonia. 22 singers, 34 normal controls, and 22 patients with hyperfunctional dysphonia where studied. In all patients, both ears were gradually masked with white noise. The change of the mean intensity level and of the mean pitch level of the speaking voice were then measured objectively with a special fundamental frequency analyzer (Fedders and Schultz-Coulon, 1975). Results show that the increase of intensity is comparable in all subjects, whereas the elevation of the mean pitch level differs significantly: trained voices (singers) react with the least pitch increment whereas dysphonic patients react with the most. The following conclusions were made from the present investigation: 1. Extreme increments in pitch level can be considered to be a more significant etiological factor of dysphonia than intensity increments; 2. Vocal therapy and voice training may have a favorable effect on the Lombard reflex (probably by improvement of the kinesthetic control mechanism) so that the speaking voice in a noisy environment is raised less with less vocal strain. The study also indicates that measurement of pitch changes during binaural masking can provide important information for the diagnosis, therapy and prophylaxis of dysphonia.


[The clinical olfactogram (author's transl)]. Clinical olfactometry is described in which patients are exposed to supraliminal stimuli of well-known types of odors. The results are then placed on a graded recognition scale as a polar coordinate system. Each clinical odor tested produces a typical recognition pattern which can easily be interpreted. Typical patterns for different conditions involving olfaction exist, and include euosmia, sensory anosmia, and respiratory anosmia. A simplified U-tube rhinomanometer can also be used, and data graphed on the same chart as for olfactometry. The procedure as described has been successfully tested on several hundred patients and controls.


Visceral and somatic reflexes produced by J pulmonary receptors in newborn kittens. The functional efficiency of J pulmonary receptor reflexes was studied in 28 newborn anesthetized kittens. J receptor stimulation was achieved by injecting phenyl diguanide (PDG) solution into the right atrium using a technique established earlier for cats. Control injections of PDG into the aorta were made. Respiration, aortic blood presure, and knee jerk were recorded. The Hering-Breuer reflex was routinely tested in each kitten by inflating the lungs through the tracheal cannula. In the newborn kitten (1-6 days old), it was not possible to elicit any effect on injection of PDG into the right atrium. At 1 wk, the visceral reflexes--apnea, rapid shallow breathing, bradycardia--could be produced by doses much higher (six times) than the adult dose. By the 10th day, the visceral reflexes were fully developed. The motor reflex, i.e., inhibition of EMG and reduction of knee jerk, was elicited after 3 wk. The earliest response was seen in a 3-wk-old kitten. All these reflexes were abolished by bilateral cervical vagotomy. These results suggest that the J receptor reflexes are poorly developed in the newborn kitten and that the motor reflexes develop much later than the visceral reflexes.


A new method to determine frequency characteristics of the respiratory system. A technique is described allowing one to determine simultaneously the resistance and reactance of the total respiratory system for various frequencies. During spontaneous breathing, regularly recurring impulses are produced at the mouth by means of a loud speaker. A Fourier analysis of the mouth pressure and flow signals yields mean resistance and reactance values, over 16 s, for all harmonics of 2 Hz up to 30 Hz. The values are in good agreement with those obtained in the absence of breathing and those determined by means of the forced oscillation technique and by body plethysmography. The reproducibility of the measurements is satisfactory (coefficient of variation: 11.6%).


An improved plethysmograph with servo control of hydrostatic pressure. One limitation of the conventional fluid displacement plethysmograph is that the hydrostatic pressure varies as recordings of volume change are obtained. Thus, use of this instrument can lead to inaccuracies that are proportional to the compliance of the biological system under study. This paper describes an improved design which utilizes a servo-control system and an open, mobile reservoir to ensure that the hydrostatic pressure is maintained constant as volume changes are recorded. The result is a device that succeeds in combining stability, sensitivity and accuracy in a fluid displacement plethysmograph.


Implantable electrode for recording nerve signals in awake animals. An implantable electrode assembly consisting of collagen and metallic electrodes was constructed to measure simultaneously neural signals from the intact nerve and bioelectrical noises in awake animals. Mechanical artifacts, due to bodily movement, were negligibly small. The impedance of the collagen electrodes, measured in awake cats 6-7 days after implantation surgery, ranged from 39.8-11.5 komega at a frequency range of 20-5 kHz. Aortic nerve activity and renal nerve activity, measured in awake conditions using the collagen electrode, showed grouped activity synchronous with the cardiac cycle. Results indicate that most of the remal nerve activity was from postganglionic sympathetic fibers and was inhibited by the baroceptor reflex in the same cardiac cycle.


Gas transport in a model derived from Hansen-Ampaya anatomical data of the human lung. The anatomical data of the human lung published by Hansen and Ampaya are used in a model of gas transport in the lung. The Bohr dead space is calculated from solutions of a transport equation where diffusivity is given by an empirical formula obtained by Sherer et al. A satisfactory agreement is found with experimental data obtained from simultaneous washouts of H2 and SF6 for respiratory frequencies ranging between 15 and 60 min-1 and tidal volumes between 200 and 1,800 ml. The results support the idea that molecular diffusion is the main but not the only physical phenomenom which intervenes in gas mixing during breathing.


Pneumotachograph for flow and volume measurement in normal and dense atmospheres. Measurement of ventilatory flow during pulmonary function maneuvers and in severe exercise by the pneumotachograph flow transducer in dense atmospheres is handicapped by the inverse relationship between the linear range of such transducers and gas density. This limitation in the usefulness of the pneumotachograph can be overcome by mounting such flow transducers in parallel and taking into account the reduction in peak ventilatory flow transducers in parallel and taking into account the reduction in peak ventilatory flow which occurs with increasing density. The design of such assemblies, their static performance characteristics, and a drift subtractor, constructed to stabilize the output of the pneumotachograph pressure transducer, are described.


Nocturnal lowering of thresholds for sweating and vasodilation. Six subjects exercised on a bicycle ergometer at 60-70% of maximal aerobic power in a 25 degrees C ambient. Experiments on each subject were conducted at night (4:00-5:30 A.M.) and in daytime (noon-4:30 P.M.). Chest sweating rate (msw) was measured with resistance hygrometry. Forearm blood flow (BF), with  an arm skin temperature of 35.5 +/- 1.2 degrees C (SD), was measured with electrocapacitance plethysmography. Esophageal temperature (Tes) was measured with a thermocouple at the level of the left atrium, and mean skin temperature (Tsk) was calculated from a weighted average of temperatures at three sites. Tes was corrected to a skin temperature of 33 degrees C as follows: T'es = Tes + (Tsk - 33 degrees C)/8. This correction reflects the relative contributions of Tes and Tsk to control of msw:T'es and BF:T'es relations were not consistently changed. In any given subject, thresholds for sweating and vasodilation were shifted about equally. These shifts averaged 0.57 degrees C (range: 0.23-0.93 degrees C)for msw and 0.63 degrees C (range: 0.17-0.98 degrees C) for BF.


Sex and age differences in pulmonary mechanics in normal nonsmoking subjects. Maximun flow-volume, static pressure-volume, and maximum flow static recoil curves of three groups of nonsmoking, normal subjects (young men, young women, elderly women) were used to assess age and sex differences in pulmonary mechanics. No significant sex differences in maximum flow were seen but the young men showed higher lung recoil pressures at full inflation. When the influence of the inspiratory muscles and chest wall was excluded by exponential extrapolation of the pressure-volume curves to a maximum volume the bulk elastic properties of the lungs of young men and women appeared identical. Loss of maximum expiratory flow at low lung volumes and of lung recoil pressure occurred with age in nonsmoking women in whom emphysema should be minimal and therefore indicate true physiological effects of aging. The changes in pulmonary mechanics with age are consistent with an increase in unstressed dimensions and loss of elastic recoil of both alveoli and airways.


Plasma renin, angiotensin II, and plasma and urinary aldosterone in running exercise. Plasma renin activity (PRA), renin concentration (PRC), angiotensin II and urinary aldosterone of four male athletes were investigated before and after a running exercise of 3 X 300 m. After the exercise, there were marked increases in all these parameters. The maximal increases (of the means and the ranges), found in the samples taken 30 min after the exercise, were: 108% (27-230%, P less than 0.05) in PRA, 490% (240-800%, P less than 0.01) in PRC, 830% (400-1,970%, P less than 0.025) in plasma angiotensin II and 1,600% (160-3,920%, P less than 0.02) in plasma aldosterone. The increase in the urinary excretion of aldosterone was 120% (42-180%, P less than 0.025). This study demonstrates that intense physical exercise may cause marked changes in all the three main components of the renin-angiotensin-aldosterone system. The significance of these changes for the physiological function of the human organism in physical stress needs further investigation.


Automic innervation of dog coronary arteries. The autonomic innervation of canine coronary arteries has been examined using the Falck and Owman technique for demonstrating catecholamines and a modification of the Koelle technique for the demonstration of cholinesterase. The experimental protocol included an examination of the neural innervation of the major coronary arteries: LCC, LAD, and RCA. A consistent, relatively dense adrenergic innervation was noted. A gradient in the degree of cholinergic innervation was: LAD  less than RCA less than LCC. Light microscopic examination of the hearts of dogs subjected to either cervical vagotomy or total extrinsic cardiac denervation was performed. Additional surgical procedures included removal of the left stellate ganglion and a preferential stripping of the LCC. These studies demonstrated the intrinsic nature of parasympathetic coronary innervation. Following all surgical procedure no variations in density of cholinergic innervation were noted, indicating that these fibers are probably postganglionic parasympathetic fibers arising from intrinsic ganglia within the ventricles. These ganglia may be located at the base of the great vessels and send their fibers to the coronary vessels via the septal artery.


Maturational effects on respiratory responses to carbon dioxide in premature infants. The respiratory responses of 18 premature infants to breathing steady-state levels of carbon dioxide were quantified by conventional minute ventilation measurements and by measuring the pressure generated after end-expiratory airway occlusions. Since no flow or volume changes occur following airway occlusion, the CO2 response obtained by this measurement is independent of vagal stretch receptor influence and mechanical changes in the lungs. When studied at 3-4 days of postnatal age, a group of 29-32-wk gestation infants had a mean slope of ventilatory response to CO2 of 10.8 +/- 5.6 ml-min-1-kg-1-mmHg-1 as compared to 53.5 +/- 10.8 ml-min-1-kg-1-mmHg-1 (P less than 0.005) for a group of 33-36 wk. When these infants were restudied at 7-10 days of age, the slopes were 21.6+/- 6.6 and 58.4 +/- 10.6 ml-min-1-kg-1-mmHg-1 (P less than 0.01), respectively. The mask pressure response paralleled minute ventilation. We conclude that the increase in ventilatory response to carbon dioxide seen with maturation is not due only to changing mechanical features of the lungs, but to increased sensitivity of respiratory centers to carbon dioxide.


Physiological aging of champion runners. The physiological adjustments to aerobic work (5.6 km/h, up a 9% grade) and to exhausting treadmill work of former champion middle-distance runners were determined in 1971, at ages 47-68 yr, 25-43 yr after their competitive careers in track. In the resting state the former athletes as a group are very much like nonathletes of the same ages. Efficiency in the aerobic walk was the same in both groups and did not change with age in either, but the former athletes on the average performed the walk with less strain as indicated by lower blood lactates, "ventilatory equivalents," and heart rates than nonathletes at corresponding ages. Mean VO2max of the runners declined from 71.4 ml/min-kg-1 in youth to 41.8 at a mean age of 56.6 yr, as compared with mean values of 50.6 and 36.5 ml/min-kg-1 in nonathletes at corresponding ages. VO2max had declined below the average of nonathletic men in only two of the former runners. Mean maximal heart rate declined with age from 186 to 180 in the runners, and from 199 to 186 in nonathletes at corresponding ages. Ventilatory responses of men in both groups were closely related to the increases of blood lactate in both aerobic and maximal work.


Systolic time intervals during +Gz acceleration. Systolic time intervals (STI) were recorded in 8 healthy male volunteers before, during, and after 30-s exposures to +3 Gz, +5 Gz, and +7 Gz acceleration. Heart rate (HR) increased at all +Gz levels, as did the HR corrected QSIc interval, left ventricular ejection time (LVETc), preejection period (PEPc) and PEP/LVET. These changes in STI were also proportional to the +Gz level. At the higher +Gz levels, PEPc and PEP/LVET continued to increase early in the recovery period, but HR and all STI returned to control after 60 s of recovery. Although physiological variables other than myocardial contractility, such as preload and afterload may influence STI during +Gz the effects of +Gz on stroke volume (SV) and cardiac output (CO) were estimated using previously described relationships between STI and invasively determined indices of cardiovascular function. In general CO increased as SV decreased. During recovery, HR and CO fell and CO remained slightly below control levels, primarily because estimated SV remained low. This study demonstrates the feasibility of using STI to estimate noninvasively the transient changes in cardiovascular function during +Gz acceleration.


Thermoregulatory response of women to intermittent work in the heat. Seven women worked intermittently in three randomly ordered sessions at 75% Vo2max at three temperatures, 28 degrees C (45% rh) 35 degrees C (65% rh), and 48 degrees C (10% rh) and recovered in a cool environment (22 degrees C) after each 6-min work period. Although Tre was higher in each successive work period, the ambient temperature had no effect on the cardiovascular or respiratory responses or on deltaTre. In all conditions SV decreased with time with a concomitant increase in HR to maintain Q. A fall in mean blood pressure from the initial to final measurement was due entirely to a decrease in diastolic pressure. The final Tre for these women was approximately equal to that previously reported for men working continuously for 1 h under conditions equivalent to time-weighted average of the thermal and metabolic loads during work and recovery in this study.


Threshold temperatures for shivering in acute and chronic hypercapnia. Threshold temperatures for shivering in acute and chronic hypercapnia were determined in guinea pigs by measuring the time course of cervical cord temperature, skin temperature, oxygen consumption (Vo2), and electrical muscle activity during cold exposure (15 degrees C). Prior to acute exposure to CO2, the shivering threshold was determined in each animal during control conditions breathing air. With increasing CO2 concentrations (5,7.5, and 15% CO2) the shivering thresholds fell to lower temperatures, decreasing by approximately 40 degrees C at 15% CO2. The shift of the shivering threshold to lower values found during acute exposure to 15% CO2 was reversed after chronic exposure to 15% CO2 for 3 days, which marks the time of metabolic adaptation to CO2.


Physiologic limits to work in the heat and evaporative coefficient for women. Ten heat-acclimated females exercised seminude on a treadmill at 30% Vo2 max (M=152 W-m-2) under eight air temperatures (Ta) ranging from 30 degrees C to 52 degrees C. Each experiment involved 1 h of fixed and a 2nd h of progressively increasing water vapor pressure (Pw) with either air movement of 1 m-s-1 or still air. The equilibrium values of rectal temperature (Tre), mean skin temperature (Tsk),and heart rate (HR) reached in the 1st h were forced upwards in the 2nd h by the rising Pw. The critical Pw was defined by the Tre inflection point for each Ta. The loci of the critical Pw were used to delineate the thermal limits on the psychrometric chart and were used to derive the effective evaporative coefficient (Ke') applicable to the ambient capacity for evaporative cooling (Emax). The derived Ke' was 17.6 +/- 4.2 W-m-2 (mean +/- SD) for v0.6m-s-1. Isotherms constructed on the basis of the obtained Ké, Tsk, and sweating capacity were higher than the physiologically based Pw limits.


Effects of body weight and food intake on pleasantness ratings for a sweet stimulus. A physiological role for pleasure has been suggested from observations that glucose loads reduced pleasantness ratings for sweet stimuli in normal weight but not in weight-reduced subjects. This hypothesis was reexamined in fasting and food-loaded, lean and obese subjects. Magnitude estimates of sweetness and pleasantness were obtained for seven sucrose concentrations. Indices of obesity including percent overweight, percent body fat, and body mass index were calculated. Pleasantness ratings of fasting subjects either increased up to a moderate concentration and then declined (type I hedonic response) or increased monotonically with concentration (type II hedonic response). Both types of response were found in lean and obese subjects. Within each weight group there appeared a negative correlation between pleasantness response and indices of obesity. Caloric loads reduced hedonic ratings of type II lean and obese subjects, but did not affect pleasantness response of type I lean and obese subjects. These findings indicate the presence of an hedonic monitor biased by body weight and caloric intake. The data suggest a physiological role for pleasure in regulating body weight at different set points in lean and obese subjects.


Effect of angiotensin on hypoxic pulmonary vasoconstriction in isolated dog lung. The effects of infused angiotensin on hypoxic pulmonary vasoconstriction in blood-perfused isolated dog lungs were studied. By using a double-perfusion system we were able to perfuse the right and left lungs separately in the same animal; one lung was used as control and the other lung was experimentally modified. The vasoconstrictive response to hypoxia decreased with time in the isolated lung preparations. The infusion of either angiotensin I or angiotensin II (1.2-5.8 mug/min) caused a threefold increase in the vasoconstrictive response to hypoxia over control levels. A second hypoxic period during the infusion usually yielded a diminished response, suggesting further degeneration of the response irreversible with angiotensin. It was concluded that angiotensin I or angiotensin II temporarily enhances hypoxic pulmonary vasoconstriction in isolated dog lungs.


Effects of limping on normal exercise response. Ten subjects performed two identical exercise tests on a motor-driven treadmill while walking normally or while limping. The two tests were separated by a rest period sufficient to permit return to a base-line state and the sugjects were randomly assigned to a normal-limp or limp-normal exercise sequence. Work rate was varied progressively by increasing treadmill grade in 5-degree increments to a maximum of 20 degrees, while limping was induced by insertion of a 0.5-cm wooden spike under the heel of the dominant foot. At all work rates, limping produced consistent increases in all variables studied, except for tidal volume where a decrease was seen. At the highest work rate these differences tended to disappear. Even considering the greater O2 consumption induced by limping, there were still significant (P less than 0.01) increases in both respiratory rate and ventilation at all but the highest work rate, accompanied by an insignificant fall in tidal volume. This suggests that these changes were not induced by the alterations in the internal or external work of walking and that limping somehow alters the regulation of exercise hyperpnea.


A digital display of neuronal average firing frequencies. A digital instrument is described which displays on a three-digit, seven-segment display the average firing frequency of a neuron. Averaging is done over essentially consecutive timing intervals of 1, 2, 5, 10, 20, or 50 s. Provision is made for overflow indication and for manual resetting. The design is simple, economical in the number of integrated circuits used, and may be adapted for measuring the average frequency of occurrence of events other than neuronal firing.


The response of plant mitochondria to media of high solute content. The state-3 rate of respiration of potato tuber mitochondria is inhibited by concentrations of KCl or NaCl above 125 mM, and by concentrations of sucrose, lactose, or maltose above 500 mM, but not at all by mannitol, glucose, glycine, or proline up to a concentration of 1500 mM in the medium. Mitochondria from cauliflower, beetroot, cucumber, rock melon, and watermelon behave very similarly to those from potato tuber. The variable response to different solutes proves that the reduction in respiration is not a simple function of the chemical potential of water in the medium. Disruption of potato mitochondria by ultrasonic vibration does not relieve the inhibition of succinate oxidation caused by KCl or sucrose. However, treatment with detergent abolishes completely the inhibition of respiration by sucrose. Inhibition of succinate dehydrogenase [Succinate:PMS, oxidoreductase (EC.1.3.99.1)] and malate dehydrogenase [L-Malate:NAD oxidoreductase (EC.1.1.1.37)] activities by sucrose is less than the inhibition of succinate- and malate-dependent oxygen uptake by the potato mitochondria. Limited substrate uptake and, alternatively, reduced electron flow as a consequence of a direct effect of solute on the mitochondrial membrane are considered as possible mechanisms of inhibition.


Phenylenediamine restoration of photosynthetic electron flux in DBMIB-inhibited chloroplasts. Phenylenediamines have been studied and compared as to their effectiveness in stimulating photosynthetic electron flux in DBMIB-inhibited chloroplasts. It has been found that N-substituted as well as C-substituted p-phenylenediamines accelerate the rate of ferricyanide reduction, a photosystem II photoreaction, under conditions where the radical cations of N-substituted p-phenylenediamines are stable. The P/e2 ratios for these partial reactions are between 0.4 and 0.5; this is taken as evidence that N-substituted p-phenylenediamines are reduced by the chloroplasts close to the outer surface. Both N- and C-substituted p-phenylenediamines are capable of bypassing the site of DBMIB inhibition and restoring electron flow from water to methylviologen. N-substituted p-phenylenediamines appear to be more effective even at high concentrations of DBMIB. The P/e2 ratios for these reactions are on the order of 0.75-1.0; this is taken as evidence that the bypass reaction for N-substituted p-phenylenediamines occurs on the inside of the thylakoid membrane.


Inhibition of phosphoribosylpyrophosphate synthesis by purine nucleosides in human erythrocytes. The effects of purine nucleosides on 5-phosphoribosylpyrophosphate (PP-ribose-P) synthesis were investigated in human erythrocytes in vitro. Previous observations have shown an increase of PP-ribose-P formation with nucleoside when inorganic phosphate (Pi) exceeded 20 mM. In contrast, when external Pi varied from 0 to 25 mM adenosine, inosine, guanosine, or 6-methylmercaptopurine riboside 1.25 mM decreased intracellular PP-ribose-P. The mechanism of this nucleoside effect on P-ribose-P concentration was investigated. Decreased synthesis rather than increased utilization accounted for the nucleoside effect. A decrease of erythrocyte Pi from the control values of 0.5 to 2.4 mM accompanied the nucleoside-related diminution of PP-ribose-P synthesis. This reduction of Pi was capable of decreasing the activity of PP-ribose-P synthetase which is known to be sensitive to small changes of Pi. The decrease of erythrocyte Pi levels was caused both by those nucleosides which were eventually degraded via purine nucleoside phosphorylase (adenosine, inosine, and guanosine) and by those nucleosides which were substantially phosphorylated initially by adenosine kinase (methylmercaptopurine riboside and adenosine with erythro-9(2-hydroxyl-3-nonyl)-adenine). Twenty-five per cent of methylmercaptopurine riboside was converted to the monophosphate derivative during the incubation. The presence of this compound, capable of inhibiting PP-ribose-P synthetase, provided evidence for a second mechanism of inhibition of PP-ribose-P synthesis by methylmercaptopurine riboside. No substantial increase of AMP, ADP, 2,3-diphosphoglycerate, or cyclic AMP nor a decrease of ribose 5-phosphate was found. These observations suggest that under physiological Pi concentrations nucleosides diminish PP-ribose-P synthesis mainly by decreasing intracellular Pi. Regulation of PP-ribose-P formation by alterations of intracellular Pi levels may be an important control mechanism and may account for a number of biological effects of nucleosides.


Determination of the subunit composition of haptoglobin 2-1 polymers using quantitative densitometry of polyacrylamide gels. Human haptoglobin (Hp), a hemoglobin-binding glycoprotein containing two types of polypeptide chains, alpha and beta, in equimolar amounts linked by disulfide bonds, exists in three major phenotypes determined by the properties of the alpha chain: Hp 1-1 (alpha1), Hp 2-2 (alpha2), and Hp 2-1 (alpha1 and alpha2). Hp 2-2 and Hp 2-1 form a series of alpha-disulfide-linked polymers. The subunit composition of the Hp 2-1 series was studied by isolation of single Hp 2-1 polymers by polyacrylamide gel electrophoresis. After reductive disulfide cleavage and alkylation the relative content of alpha2 and alpha1 polypeptide chains was determined by quantitative densitometry of acid/urea polyacrylamide gels stained with Coomassie brilliant blue R250. The molar ratios alpha2/alpha1 for the Hp 2-1 polymers. P1 through P5 (in order of decreasing electrophoretic mobility), were found to be: P1, 0.0 (alpha1 only); P2, 0.48; P3, 0.97; P4, 1.6; P5, 2.0. Since one alphabeta-Hb half-molecule is known to bind to each Hp beta chain, the beta polypeptide chain content of each of the Hp 2-1 polymers could be estimated by by counting the number of Coomassie blue bands formed after electrophoresis of isolated Hp 2-1 polymers fractionally saturated with cyanmethemoglobin (Hb). The number of beta chains present in Hp 2-1 polymers P1 through P4 was determined to be: P1, 2; P2, 3; P3, 4 and P4, 5. Molecular weights of the Hp 2-1 polymers were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresus using as standards the almost homologous Hp 2-2 polymer series whose molecular weights are known from ultracentrifugation studies. Molecular weights for the first five Hp 2-1 polymers were estimated to be 107,000; 162,000; 217,000, 274,000; and 331,000, respectively. These data are consistent with the previously proposed model for the subunit composition of the Hp 2-1 polymer series when P1 = (alpha1 beta)2 and the subsequent polymers in order are represented as (alpha1 beta)2(alpha2 beta)n where n = 1,2,3,4...


Structure of the accumulating oligosaccharide in fucosidosis. Two water-soluble complex carbohydrate storage products were isolated from tissues and urine of patients with an inherited deficiency of lysosomal alpha-L-fucosidase (fucosidosis). The major component was an oligosaccharide of approximate molecular weight 1700, indicating that it was a dekasaccharide. From a combination of sequential digestion with purified exo-glycosidases, periodate oxidation and permethylation in conjunction with gas-liquid chromatography mass spectrometric analysis, the structure was found to be: Fuc(alpha 1 leads to 2)Gal-(beta 1 leads to 4) GlcNAc (beta1 leads to 2)Man [Fuc(alpha1 leads to 2) Gal (beta1 leads to 4) GlcNAc(beta1 leads to 2) Man] (alpha 1 leads to 3/6) Man (beta1 leads to 4) GlcNAc, although there was some evidence for heterogeneity at the mannose branchpoint. This material is structurally related to the stored oligosaccharides in patients with inherited deficiencies of beta-galactosidase (G M1-gangliosidosis) and N-acetyl-beta-hexosaminidase (G M2-gangliosidosis). A dissaccharide with the probable structure Fuc(alpha1 leads to 6)GlcNAc was found in lesser amounts in tissues; both are believed to be derived from the impaired catabolism of large numbers of different glycoproteins.


Concanavalin A binding to membranes of the Golgi apparatus and resultant modification of galactosyltransferase activity. 125I-Concanavalin A was found to bind to purified Golgi membranes in a manner which was time-dependent, proportional to the amount of Golgi protein, saturable, and inhibited by alpha-methyl-D-glucoside and unlabeled concanavalin A. Approximately 2 nmol of 125I-concanavalin A were bound per mg of membrane protein with a mean Ka of 0.2 x 10(7) M-1. After binding of concanavalin A to the membranes, all unbound lectin was removed by repeated washing prior to assay of galactosyltransferase activity to assure that bulk concanavalin A would not bind to the sugar acceptor during the enzyme assay. Therefore, the effects of the lectin were not due to altered acceptor concentrations in the assay. As a result of concanavalin A binding the activity of the galactosyltransferase of these membranes was enhanced by 70 to 100%. The enhancement was abolished by treatment of the concanavalin A-bound Golgi with alpha-methylglucoside. Kinetically this stimulation was reflected as an altered Km for N-acetylglucosamine and UDP-galactose. The data are suggestive that the stimulation of the enzyme results indirectly from perturbation of the membranes on binding of the lectin.


Poly (A)-rich ribonucleoprotein complexes from HeLa cell messenger RNA. Polyribosomal messenger RNA from HeLa cells contain 3'-OH-terminal polyadenylate sequences approximately 133 nucleotides in length (weight average). When analyzed at the ribonucleoprotein level of organization these poly(A)-rich sequences are found to contain tightly bound proteins. These proteins remain associated with the poly(A)-rich RNA during affinity chromatography of RNase A and T1-digested polyribosomes on poly(U)-Sepharose in 0.5 M NaCl, and co-elute from the column with the RNA at 50% formamide. Controls establish that the co-purification of the proteins with poly(A) on poly(U)-Sepharose requires the molecular integrity of the poly(A). Polyacrylamide gel electrophoresis resolves the poly(A)-specific proteins into two components of 74,000 and 62,000 molecular weight. The larger protein is the same size as that previously reported to be associated with poly(A)-rich sequences in HeLa heterogeneous nuclear RNA (Kish, V.M., and Pederson, T. (1975), J. Mol. Biol. 95, 227-238). It is concluded that both HeLa nuclear and polyribosomal poly(A) sequences have a protein (62,000 molecular weight) associated with poly(A) appears to be confined only to messenger RNA.


The NH2- and COOH-terminal amino acid sequence of nuclear protein A24. The NH2- and COOH-terminal sequence of nuclear portein A24 has been determined by automatic Edman degradation and carboxypeptidase A and B digestion. Protein A24 is of interest because it is composed in part of histone 2A (Goldknofp, I.L., and Busch, H., (1975) Biochem, Biophys. Res. Commun. 65, 951-960). The sequence of the first 37 NH2-terminal residues is: Met-Gln-Ile-Phe-Val-Lys-Thr-Leu-Thr-Gly-Lys-Thr-Ile-Thr-Leu-Glu-Val-Glu-Pro-Ser-Asp-Thr-Ile-Glu-Asn-Val-Lys-Ala-Lys-Ile-Gln-Asp-Lys-Glu-Gly-Ile-Pro- This sequence is not homologous to any known histone sequence. It contains regions of internal homology (italics). The COOH-terminal amino acid sequence is the same as that of histone 2A, naely: -His-His-Lys-Ala-Lys-Gly-Lys-COOH.


Purification and characterization of two lectins from Caragana arborescens seeds. A glycoprotein fraction with hemagglutinating activity was purified by affinity chromatography from seeds of the pea tree, Caragana arborescens. Subsequent fractionation resolved two components, which could be separated on a preparative scale using different affinity matrices. The major component binds to N-acetylgalactosamine coupled to Sepharose 4B. It is a glycoprotein with high hemagglutinating activity. It is composed of two types of polypeptides, present in nonstoichiometric amounts, with apparent molecular weights near 30,000. In the native molecule, the subunits are cross-linked by disulfide bonds to form dimers, which in turn appear to be in rapid equilibrium with tetramers. The minor component binds to underivatized Sepharose 4B. It too, is a glycoprotein but has low hemagglutinating activity. It is composed of three types of polypeptides which, although they have apparent molecular weights near 30,000 are distinguished from the subunits of the major hemagglutinin by a number of physical and chemical properties. The native molecule is dimeric, with a mass of 60,000 daltons. The major component has high affinity (K = 0.1 mM) for the haptenic sugar, N-acetylgalactosamine, but will also bind D-galactose. Neither lectin has ABO blood group specificity, nor are they transformed mouse fibroblasts to the same extent.


Electronic structure and bonding of the amino acids containing first row atoms. The electronic structures of the amino acids containing first row atoms have been determined for the zwitterionic form using an approximate self-consistent field method, partial retention of diatomic differential overlap. Various energetic quantities including certain proton affinities are present as are eigenvalues for the highest occupied and lowest unoccupied molecular orbitals. It is found that our method, in common with all methods employing minimum basis sets, yields eigenvalues for the highest occupied molecular orbital that are too high. The method does predict the location of this orbital correctly when compared to calculations employing larger basis sets. It is predicted that electron loss due to ionizing radiation should occur from the carboxylate group for the nonaromatic amino acids, while for tyrosine and tryptophan, electron loss should occur from the ring system. No choice between these two sites can be made for phenylalanine. Charge distributions have been obtained which show that only partial zwitterionic character is found in the backbone and that little delocalization of charge from the backbone to the side chain occurs. Localized molecular orbitals have been obtained using the Boys criteria and the bonding in the amino acids is disscussed in terms of these orbitals. Hybridization of various bonds and bond polarities are discussed as is the phenomenon of fractional bonding to carbon.


Maternal-fetal carnitine relationship and neonatal ketosis in the rat. The concentration of ketone bodies in plasma and of carnitine in various maternal, fetal, and neonatal tissues was examined during the developmental period in rats. Plasma ketone levels were low in the fetus, increased 10-fold during the first 24 h postpartom, and thereafter gradually declined such that normal values were found at the end of the suckling period. An almost identical profile was observed for liver carnitine concentrations in the baby rats. The converse was true for heart tissue, the carnitine content of which was low at birth and steadily increased to adult levels with the time of suckling. The primary source of carnitine in neonatal tissues, at least during the first 2 to 3 days postpartum, was shown to be the mother rat whose liver and milk carnitine content was very high at this time and fell as nursing continued. Experiments in which the fate of [14C]butyrobetaine, the immediate precursor of carnitine, was followed after injection into nursing mother rats indicated movement of carnitine from maternal liver leads to maternal plasma leads to milk leads to neonatal tissues. The above findings support the view expressed earlier that one prerequisite for the development of a high ketogenic profile in liver may be an elevation in the tissue carnitine concentration. Additional factors, however, are clearly involved as evidenced by the observation that in the fed state perfused livers from nursing mother rats synthesized ketone bodies from oleic acid at low rates compared with those seen after a 24 h fast, despite the fact that tissue carnitine levels were equally elevated in both groups. This paradox is likely related to the fact that in the fed state such livers also contained large quantities of glycogen, depletion of which through fasting was accompanied by marked acceleration of ketogenesis from oleate. The data indicate, therefore, that maximal ketogenic capacity of the liver requires for its induction an increase in carnitine coupled with a decrease in glycogen content of the tissue.


Effect of glycosylation on the in vivo circulating half-life of ribonuclease. The circulating half-lives of the four isozymes of bovine pancreatic ribonuclease (RNases A, B, C, and D) have been determined in normal and in nephrectomized rats. The isozymes differ only in their glycosyl content. While A contains no sugars, B has a simple oligosaccharide (GlcNAc2 Man4-5),and C and D each have a complex oligosaccharide (GlcNAc4 Man 2-3 Gal2 Fuc NeuAc2, and GlcNAc4 Man3 Gal2 Fuc NeuAc4, respectively) attached to Asn-34 of the polypeptide chain. All four isozymes were cleared rapidly in normal rats (t 1/2 = 2 to 3 min), as expected on the basis of the established role of the kidneys in removing low molecular weight proteins from circulation. In nephrectomized rats, however, a much slower clearance was observed, thus permitting the evaluation of the role of the carbohydrate chains in the catabolism of the isozymes. The clearance curves can be analyzed in terms of two processes, a rapid initial one, shown to represent the equilibration of the injected enzyme into extravascular space, and a second one which is interpreted as the catabolic clearance of the enzyme. The haf-life of the RNase isozymes was calculated from this second process and found to be in the range 528 to 577 min for RNase A, 15 min for RNase B, 681 to 862 min for RNase C, and 839 to 941 min for RNase D. The rapidly cleared RNase B was treated with alpha-mannosidase to remove 3 of the 4 mannosyl residues, leaving only a trisaccharide (GlcNAc2-betaMan) attached to the protein. The half-life of this RNase B derivatives was found to be in the range 616 to 733 min. From these results it is concluded (a) that the addition of complex oligosaccharides to a protein does not have any significant direct effect on its circulating half-life (RNases C and D compared to RNase A), and (b) that in the rat there exists a mechanism for clearing glycoproteins based on specific recognition of exposed alpha-mannosyl residues (RNase B compared to the other isozymes and to alpha-mannosidase-treated RNase B).


Structure and action of heteronemertine polypeptide toxins. Primary structure of Cerebratulus lacteus toxin B-IV. The amino acid sequence of Cerebratulus toxin B-IV, a crustacean-selective axonal toxin occurring in the marine worm C. lacteus, was determined by Edman degradation of the tryptic and staphylococcal protease peptides obtained from the reduced, carboxymethylated toxin. All four of the anticipated maleylated tryptic peptides, ranging in size from 8 to 23 residues, and three staphylococcal protease peptides, ranging from 9 to 35 residues, were isolated in pure form by gel filtration followed by either ion exchange chromatography or preparative paper electrophoresis. The order of the maleylated tryptic peptides was based upon the sequences of the staphylococcal protease peptides. As might be expected, toxin B-IV displays no homology with the elapid nicotinic receptor toxins. In addition, toxin B-IV is structurally unrelated to a group of scorpion neurotoxins which, like B-IV, affect action potential generating mechanisms.


Preparation of intact monomeric collagen from rat tail tendon and skin and the structure of the nonhelical ends in solution. Procedures for the preparation of soluble collagen from rat skin and tail tendon were reviewed and revised to permit the preparation of native monomeric collagen with intact nonhelical ends. The degree of intactness was estimated from the tyrosine content, which is present only in the nonhelical ends, and by mobility of the COOH-terminal cyanogen bromide peptide of the alpha1 chain on sodium dodecyl sulfate gels. The amount of covalently cross-linked polymeric material present was estimated by molecular sieve chromatography of denatured samples. Rapid purification in the cold was sufficient to prevent or greatly reduce proteolytic alteration. Fractionation by salt precipitation at acid pH was effective in reducing the content of polymeric material. Rat tail tendon yielded completely intact native collagen, but some high molecular weight aggregates remained. Collagen from the skin of lathyritic rats was easier to obtain free of aggregates, but contained about 1 less tyrosine residue per alpha1 chain even when isolated in the presence of enzyme inhibitors. Proton NMR spectra of denatured acidic solutions of these preparations showed that 4 to 5 tyrosine residues per alpha chain were present, confirming the chemical analysis. Spectra of the native molecule showed that about the same number of tyrosine residues per chain are in rapid motion, unlike residues in the helical portion of the molecule, a result which shows that the nonhelical ends of the native molecule are unstructured in acidic solution.


Absence of the major sialoglycoprotein in the membrane of human En(a--) erythrocytes and increased glycosylation of band 3. The human En(a-) blood group is a rare recessive trait. These erythrocytes lack the major membrane sialoglycoprotein (PAS1, MN protein, or glycophorin) and contain a decreased amount of PAS2 as demonstrated by radiolabeling of surface porteins and chemical techniques. A third glycoprotein, Band 3, contains two labeled oligosaccharide chains; the more complex oligosaccharide has a higher molecular weight in En(1-) cells than in normal cells. A fourth glycoprotein, PAS3, is present in usual amounts in En(a-) cells. Cells heterozygous for En(a) are intermediate in these respects. The sialic acid is decreased in En(a-) cells, but the total carbohydrate is similar in the different membranes. The glycolipids are present in normal amounts but are much more exposed to galactose oxidase in En(a-) cells than in normal cells.


Deuterium magnetic resonance spectroscopy of isotopically labeled  mammalian cells. Choline completely deuterated in the methyl groups has been incorporated into two mammalian cell systems. The first was the chemically transformed mouse fibroblast LM cell line, grown in suspension culture. The second system consisted of mitochondria from Sprague-Dawley rats which were fed upon a choline-deficient diet enriched with deuterated choline chloride. Two classes of deuterium nuclear magnetic resonance (NMR) signal were obtained from each system. The first class of signal exhibited no residual quadrupole coupling constant and was assigned to the natural abundance of 2H in water, together with free labeled choline. The second class of signal had a residual quadrupole coupling of about 1 HKz and was assigned to 2H-labeled choline headgroups of phospholipids. These results are in conflict with a previous study of deuterated rat liver mitochondrial membranes. Deuterium magnetic resonance spectra can be obtained on mammalian membranes that have incorporated specific deuterium-labeled compounds and, therefore, provides a very powerful method for studying the dynamic structure of membranes.


The repair of the surface structure of animal cells. Experiments were performed to determine if animal cells in culture possess specific mechanisms to repair surface molecules damaged by enzymes. The surface membranes of a primary cell culture, chick fibroblasts, a permanent hamster cell line, BHK21/C13, and its virally transformed counterpart, C13/B4 were damaged by exposure to trypsin or to neuraminidase. Following digestion with trypsin, the incorporation of radioactive amino acids or sugars into purified surface membrane of cells was monitored. No differences were noted in rates of incorporation when control and trypsin-damaged cells were compared. Neuraminidase damage to the surface of BHK21/C13 and C13/B4 cells was evidenced by altered gel filtration profiles of surface glycopeptides, i.e., delayed elution because of reduction in size. By labelling cells with 14C-L-fucose prior to neuraminidase treatment and following the incorporation of 3H-L-fucose into cell surface glycopeptides after neuraminidase digestion, we were able to monitor the synthesis and turnover of fucose-containing glycopeptides in the same cells. Gel filtration profiles indicated that little or no desialylated glycoproteins were resialylated (repaired) by specific replacement of sialic acid. Comparing neuraminidase-digested and control cells we observed no difference in rates of 3H-L-fucose incorporation or of 14C-L-fucose loss from these cells; nor did we find differences in the rate of incorporation of isotopic glucosamine into sialic acid. Neuraminidase treatment failed to alter the rate of cell growth or the pattern of isotopic incorporation into various cell surface components. These results support the suggestion that return of sialic acid (repair) was effected by turnover which serves as a non-specific repair mechanism to replace damaged cell surface molecules (Warren and Glick '68; Warren, '69).


Variation in melanosome numbers in cultured B-16 melanoma cells. Melanosomes from B-16 mouse melanoma cells in culture were isolated by treatment of pigmented cells with 2% SDS, sonication, and heating at 100 degrees C. The total number of melanosomes in cultures of B-16 mouse melanoma cells increased exponentially during the rapid phase of sigmoid growth. The numbers of melanosomes per cell decreased during rapid phase of growth, and repigmentation was observed only when the cultures attained the stationary growth phase. BUdr at a minimum concentration of 0.5 mug/ml decreased both cell growth and numbers of melanosomes per cell, and completely inhibited repigmentation following a period of active growth. Cells cultured in 0.1 mug/ml BUdr grew at the same rate as untreated cells but contained fewer melanosomes/cell and lower total numbers of melanosomes during the late stages of the growth cycle.