Sorbitol and other polyols in lens, adipose tissue and urine in diabetes mellitus.

Sugars and polyols (glucose, fructose, sorbitol, inositol and pentitols) have been measured in urine, adipose tissue and lens in groups of diabetic and non-diabetic patients. The daily excretions of glucose, sorbitol and inositol were raised in the diabetics. There was a linear relation between 24 hour urinary excretion of glucose and hexitols (r = +0.87, p less than 0.001). In lens of diabetics there was an increase in glucose concentration, but not of fructose, sorbitol or inositol. Compounds readily detected in adipose tissue were inositol and glucose; in addition an unidentified carbohydrate was detected in adipose tissue that related to the concentration of tissue glucose. These results are discussed in relation to the possibility that tissue accumulation of polyols could be responsible for the secondary complications of diabetes such as cataracts.


Prenatal diagnosis of fucosidosis.

Pregnancy from a family at risk for fucosidosis was monitored. The fetus was diagnosed as having a carrier state of the disease. alpha-L-fucosidase activity, however, was found to be absent in white blood cells obtained from identical twins after delivery. The diagnostic evaluation of (1) the enzyme activity in amniotic fluid and in cultivated amniotic fluid cells and (2) the presence of fucose rich compound in amniotic fluid are discussed.


Adherence of rabbit erythrocytes to guinea-pig brain structures. Evidence for antigenic correlation between guinea-pig thymus and brain?

Similar to thymocytes in the thymic cortex, certain brain structures, such as nerve tracts and nerve cells of the Ammon's horn, show an affinity to rabbit erythrocytes (RRBC) which is exemplified by the adherence of RRBC to frozen brain sections. The experimental conditions of this adherence reaction are identical with that previously found with guinea-pig thymus and lymph node sections. From the results it may be concluded that receptor substances similar in nature on thymocytes and brain are responsible for this adherence phenomenon.


Intensive immunosuppression in patients with disseminated sclerosis. III. Lymphocyte response in vitro.

Lymphocytes from fifteen multiple sclerosis patients gave responses to phytohaemagglutinin (PHA), conconavalin A (con A) and pokeweed mitogen (PWM) which were in the normal range. However, the responses of lymphocytes to stimulation by an allogeneic lymphoid cell line (LCL) were significantly lower in HLA-7-positive than in HLA-7-negative patients (a distinction not found in control groups). Depression of con A, PHA and PWM responses were observed during intensive immunosuppression. Responses to LCL were unaltered or increased during initial azathioprine and prednisone treatment. The depression of this response following antilymphocyte globulin (ALG) treatment was delayed in the HLA-7-positive patients. One week after the end of ALG treatment, most PHA, con A and PWM responses had returned to low normal values. Reduction of azathioprine and prednisone treatment at the end of 1 year resulted in a sharp rise in PHA and con A responses in some patients. Relapses in patients were frequently associated with low responses to LCL cells.


Delayed cutaneous hypersensitivity reactions to concanavalin A-binding tumour membrane components.

The use of affinity chromatography has permitted the isolation of those tumour membrane fractions possessing affinity for the lectin concanavalin A. That these fractions are rich in tumour-associated antigens is supported by their ability to induce delayed cutaneous hypersensitivity reactions. The tumour extracts possessing concanavalin affinity resulted in skin test reactivity of considerably greater frequency and magnitude than normal tissue fractions, disrupted unfractionated tumour membrane extracts, or tumour membrane fractions not possessing concanavalin A affinity. Although the present data does not permit the correlation of skin reactivity with patient or disease parameters, the isolation of augmented concentrations of tumour-associated antigens may lead to improved diagnostic and prognostic tests and vaccines for patients with cancer.


Tissue blood flow and oxygen transport in critically ill patients.

The theoretical and practical solutions to the problems of increasing oxygen transport are well understood. Unfortunately the quantitation of hypoxia, both as an absolute deficit and as a precise method of prognosis is not yet available. This may well be because regional hypoxia in a vital tissue cannot be mirrored in a total body measurement. In the low-flow state, oxygen delivery can be maintained by redistribution of cardiac output, reduction of oxygen uptake by ischemic tissue by reducing work load, by increasing oxygenation of the blood, or by decreasing the affinity of oxygen for hemoglobin. The latter provides for more oxygen to be delivered by a given amount of oxyhemoglobin before the tension falls to deleterious regions (about 20 torr). There is some evidence that pharmacologic doses of methylprednisolone may be beneficial in this respect.


Reduction in colloid osmotic pressure associated with fatal progression of cardiopulmonary failure.

The relationship between colloid osmotic pressure (COP) and the severity of cardiopulmonary failure was investigated in 99 consecutive patients admitted to our Shock Unit including 41 survivors and 58 fatal cases. The COP was significantly lower (p less than 0.001) in fatal cases in comparison to survivors. None of 21 patients in whom COP was less than 10.5 torr survived. A progressive increase in survival with typical S-shaped distribution was observed as COP increased from 10 to 19 torr, above which all patients survived. These preliminary observations demonstrated that reduction in COP in critically ill patients is associated with increases in mortality. The mechanisms by which lowering of COP may be related to fatal progression of cardiopulmonary failure is the subject of continuing study.


Algorithm for resuscitation: a systematic plan for immediate care of the injured or postoperative patient.

A systematic integrated approach to the diagnostic, monitoring and fluid volume therapy was developed for use in patients with accidental and elective surgical trauma. An algorithm (patient care protocol) is proposed for expeditious resuscitation in emergency situations using: (a) BP as the criteria for initiation of rapid fluid therapy, (b) hematocrit for the choice of blood transfusion or plasma expanders, and (c) CVP, urine output, arterial pressure and wedge pressure as criteria for slowing down or stopping the rate of volume therapy. History, physical examination, laboratory work, X-rays, monitoring and diagnostic procedures are interdigitated in a systematic fashion according to priorities of the most common life-threatening aspects of the total resuscitation problem. In chaotic emergency situations, it is impossible to plan for all possible contingencies; to try to do so results in an impossibly complex and unwiedly plan. However, we believe that almost any reasonable plan is better than no plan at all.


Assessment of intra-aortic balloon counterpulsation in cardiogenic shock.

Intra-aortic balloon pumping (IABP) has been shown to reverse the cardiogenic shock syndrome, but the long-term results (16 to 53% survival) have, in general, been disappointing. The most productive areas for the use of IABP are in the treatment of acute myocardial ischemia where in one report of 16 patients treated with IABP and surgery 15 survived and 13 were angina free, and in the open heart surgical setting where survival ranges from 42 to 70%. The implications of these treatment modalities are discussed.


Effect of methylprednisolone on predicted myocardial infarction size in man.

Therapeutic manipulations designed to conserve myocardium in patients with acute myocardial infarction appear to improve prognosis. To assess the role of glucocorticoids given in pharmacologic dosage in the early treatment of patients with acute myocardial infarction, serial serum creatine phosphokinase (CPK) were obtained every 1-2 hours in 39 consecutive patients admitted with acute myocardial infarction. Determination of completed infarction size (ISc) was made using all available CPK values (range 70-160 hours). Predicted infarct size (ILp) was based on early data following the rise in CPK from baseline values: projected CPK values were obtained over a 160 hour period using a curve fitting procedure based upon nonlinear Gauss-Newton stepwise iterations. In 13 uncomplicated control patients ISp was 43.2 +/- 11.6 (mean +/- SE) CPK-gram-equivalents (CPK-q-eq), based on data from the first 7 hours following the rise in serum CPK, while ISc was 44.7 +/- 11.4 CPK-geq (r = .99, n = 13). In 7 additional control patients whose hospitals courses were complicated by clinical extension ISp was 71.8 +/- 18.0 CPK-g-eq while ISc was 118.6 +/- 31.0 CPK-g-eq (p less than .03). In 19 patients treated with 3 grams of methylprednisolone 7-14 hours following the rise in serum CPK from baseline, data from early CPK determinations (7 hours) indicated an ISp of 118.5 +/- 24.1 CPK-g-eq while total CPK data indicate an ISc of 89.6 +/- 13.2 CPK-g-Eq (p less than .04). The exponential clearance of CPK (kd) was approximated in the controls (kd = .00095 +/- .00007 min-1) and glucocorticoid treated patients (kd = .00099 +/- .00006 min-1) and found to be similar. Thus, administration of a pharmacologic dose of methylprednisolone 7-14 hours following rise in serum CPK from baseline in a group of patients with acute myocardial infarctions has resulted in salvage of myocardium.


Second annual SCCM lecture. The role of hemodynamic monitoring in the management of the critically ill.

Balloon flotation catheterization of the central circulation provides data which may be most meaningful and important to the management of critically ill patients. It allows the measurements of the filling pressures of the right and left ventricle as well as the cardiac output. These data combined with information concerning cardiac rhythm, heart rate, arterial pressure and other variables place the principal determinants of cardiac function at the disposal of the critical care personnel. In particular, the physician may optimize left ventricular filling pressure and then after other functional determinants of cardiac performance so as to optimize the relationship of cardiac output to the metabolic needs of the body tissues and systmes. Continued hemodynamic monitoring also provides prompt objective information as to success or failure of therapeutic interventions. It thereby allows for the most rational decision making and effective alteration in the therapy applied to the individual critically ill patient. Much still needs to be done to establish completely the impact of such monitoring techniques upon patient management. However, experience thus far has been most promising. Large numbers of physicians and health care personnel now rely on pressures provided by the pulmonary capillary wedge pressure in their decisions about fluid therapy. Others are rapidly assessing the importance of the cardiac output and other parameters of cardiac function on a semi-continuous basis. Thus, balloon flotation catheterization has allowed the application of sound physiological principles to the understanding of circulatory abnormalities characterizing important patient illnesses and provides a rational basis for the selection of therapy with objective quantitative assessment of responses. The procedures are simple: the complications rate is low, and the information provided is highly relevant to clinical practice.


Symposium: Procidentia of the rectum: teflon sling repair of rectal prolapse, Lahey Clinic experience.

The Teflon-sling method of repair of rectal prolapse in the Lahey Clinic experience has proved to be one of no mortality and low morbidity, with a recurrence rate of 7.3 per cent over an average follow-up period of nearly four years. Bowel management and incontinence are problems inherent in the pathogenesis of the problem and, though improved, necessitate long-term patient re-education and physiotherapy. More than 85 per cent of the patients were satisfied with the results of the procedure.


Leiomyosarcoma of the colon: report of a case and analysis of the relationship of histology to prognosis.

A case of primary leiomyosarcoma of the colon in a patient without gastrointestinal symptomatology who initially manifested a metastasis in the parotid region is described. Review of the literature indicates that mitotic activity is the most reliable histologic criterion for prognosis. Three of five patients with tumors manifesting high mitotic activity succumbed to widespread metastatic disease within four months of initial diagnosis. The prognosis is generally favorable for those patients whose neoplasms have low or moderate mitotic rates.


Perianal verrucose carcinoma spreading to the rectum: report of a case.

The case of a 39-year-old man with a perianal verrucose carcinoma of 12 years' duration is presented. After local resection the tumor recurred several times and spread to the rectum. An abdominoperineal resection revealed neither infiltration of deeper layers nor lymph-node metastasis. Five other cases of perianal verrucose carcinoma, only two of which were described in detail, have been reported. Probably other examples have been reported under the names of "perianal florid papillomatosis,"13,23 "giant condyloma acuminatum"26 and "condyloma acuminatum with malignant transformation."9,12,27


Anal fibrosarcoma: report of a case and review of literature.

A unique case of anal fibrosarcoma is reported. Review of the literature in the past half century revealed only 13 cases of rectal fibrosarcoma, and no case of anal fibrosarcoma. Abdominoperineal resection is the usual treatment if the disease is confined to the rectum or anus. An extended resection may be indicated in those patients who have more extensive disease. Long-term follow-up information was not available in the literature. The prognosis is presumed to be the same as if not worse than for fibrosarcomas in other parts of the body. Reporting of this rare tumor when found in unusual locations is encouraged.


Ulcerative disease of the colon proximal to partially obstructive lesions: report of two cases and review of the literature.

Carcinoma complicating idiopathic ulcerative colitis is well known. Conversely, acute colitis complicating obstructing carcinomas and other partially obstructing lesions of the colon has not been recognized until recently. The present study reports two cases of colitis secondary to obstruction: 1) a giant ulcer with colitis proximal to partially obstruction diverticulitis of the sigmoid colon, and 2) colitis proximal to obstructing carcinoma of the sigmoid colon. The purpose of this report is to document these cases and review the literature on this variety of colitis to facilitate its recognition and subsequent correct treatment. An unawareness of this entity prejudices the anastomosis and results in anastomotic complications (approximately 25 per cent), with significant morbidity and mortality.


Present situation of schistosomiasis in the Libyan Arab Republic. I. Bilharziasis in Ubari District in Fezzan (Sebha Governorate.

A survey for urinary schistosomiasis was conducted in all the schools (18 in number) of ubari district (Sebha Governorate) of the Libyan Arab Republic. Out of 1874, school pupils examined 99 proved positive at the rate of 5.3 percent. Boys were more infected than girls and the peak of infection was at eleven years. The intensity of infection was light. New foci were detected in five towns and villages brignging up the total number to 20 out of 31. Samples of population in nine towns and villages (about one quarter) were examined covering 1435, with 34 infected at a rate of 2.4 percent. Treatment of infected pupils started with Niridazole at the standard dose, with about 60 percent cure rate. Eleven towns and villages were found to harbour Bulinus snails in some of their farms, increasing the number to 20 out of 31. These snails belong to subgenus Bulinus. However, they fall in four distinctive types based on the variation of shell morphology. Antisnail measures were introduced for the first time using copper sulphate with satisfactory preliminary results.


Cutaneous schistosomiasis.

Cutaneous manifestations of schistosomiasis can be produced by human and nonhuman species. They can occur in the invasive stage or oviposition stage. Manifestations in the invasive stage are non-specific and include: 1. severe itching 2. Generalized anaphylactiod reaction with urticarial or erythema multiforme-like eruption. Manifestations in the stage of oviposition are specific and include: 1. Genital and perigenital granulomata; 2. Extragenital cutaneous schistosomal granulomata occurring in sites away from the portocaval anastomoses. The clinical and the histopathologic picture are described. The possible mechanisms of ectopic localisation of schistosomal granulomata are discussed.


Control of snail hosts of bilharziasis in Egypt. 2. Effect of triphenyltin hydroxide (Du-Ter) on carbohydrate metabolism of the snails Biomphalaria alexandria and Bulinus tuncatus.

Previous work has shown that the aerobic oxidation of certain intermediates of Krebs cycle by the snail B. alexandrina and B. truncatusis inhibited by TPT. This paper reports data on the effect of TPT on glucose utilization, glycolysis, glycogen content, and glucone ogenesis in snail tissue preparations. The compounds at its LC50 inhibited gluconeogenesis, stimulated glycolysis and markedly reduced the glycogen content and glucose utilization in both snails. However, the effects were more pronounced in B. truncatus than in B. alexandrina. Possible interpretations of these findings are discussed with reference to published arrangements for regulation of gluconeogenesis and glycolysis, coupling of electorn transport to ATP synthesis and also to our present knowledge of the chemical and biological specificity of TPT.


Lipid pattern in Bilharzial hepatic fiprosis.

Plasma total lipid phosphorus, individual phospholipid fractions, free and esterified cholesterol, triglycerides and nonesterified fatty acids were fractionated by thin-layer chromatography and determined in bilharzial hepatic fibrosis patients (57 cases) and normal controls (25 cases). Significant diminutions were encountered in the total and the individual phospholipids, especially the lysolecithin fraction in the 3 groups of patients studied. Total and esterified cholesterol and triglycerides showed significant decreases, especially in moderate and late cases. NEFA, on the other hand, did not reveal any change from normal level in either moderate, late or mixed cases. Such findings may be attributed to malabsorption, lack of energy, insufficiency of required precursors as well as impaired synthesis by the liver.


Anaerobic recovery in man.

The lactic acid (L.A.) concentration in blood after a 20 sec supramaximal exercise (2.5 times VO2max) has been measured in 4 subjects in the following experimental conditions: a) during the resting period following the supramaximal exercise (rest recovery) and b) during a 3 min exercise at VO2max immediately following the supramaximal effort (exercise recovery). The L.A. concentration in blood has been found to be consistently higher (on the average by 16.9 mg%) in case (b). Since in such condition it may be reasonably assumed that the oxygen taken up by the subject is completely utilized for the exercise, the increase of blood lactate is considered evidence for the occurrence of anaerobic recovery, i.e. of a partial re-synthesis of the high energy phosphate stores of the muscle (GP = ATP + PC) depleted during the supramaximal effort, at the expense of anaerobic glycolysis. From the increase in blood L.A. concentration during the anaerobic recovery period, the amount of L.A. produced has been estimated together with the amount of GP resynthesized. The latter corresponds to 4 to 7 mMoles/kg of muscle, i.e. to about 25% of the average GP concentration in resting human muscle. The finalistic implication of this mechanism is the prompt restoration of the potential maximal power of the muscle even in the absence of O2.


Skeletal muscle leucine incorporation and testosterone uptake in exercised guinea pigs.

We examined the changes induced by daily treadmill exercise on body weights, plantaris muscle weights, plantaris protein concentrations, and L-leucine-4,5-3H incorporation into plantaris muscles of normal and castrated young male guinea pigs and of castrated animals receiving testosterone replacement therapy, and compared the testosterone-1,2-3H uptake by plantaris muscles of trained normal guinea pigs to that of untrained animals. Trained animals exhibited significantly lower body and muscle weights and greater labeled leucine incorporation into sarcoplasmic and myofibrillar proteins but did not show significant changes in protein concentrations or labeled testosterone uptake. The level of physical activity of the young animals studied appeared to be more important than gonadal endocrine function in altering protein metabolism and muscle and body weights. Because hypertrophy did not occur in the trained plantaris muscles, which had elevated rates of labeled leucine incorporation, it appears that the trained animals had a higher muscle protein turnover rate. It seems unlikely that testosterone plays an important role in these activity-related phenomena.


Ammonia production following maximal exercise: treadmill vs. bicycle testing.

From a population of 20 healthy male volunteers, half performed constant speed, incremental load maximal aerobic capacity (VO2max) tests on a motor-driven treadmill, while the other half performed similar VO2max tests on a bicycle ergometer. The two groups, matched for size and age, showed no significant differences in VO2max, maximum heart rate, or in post-exercise (4 min ) peripheral venous blood concentrations of lactete or pyruvate. However, post-exercise peripheral venous blood ammonia levels were significantly higher in the group tested on the bicycle ergometer than in the treadmill group.


Acute blood biochemical alterations in response to marathon running.

Adrenal-sympathico function, blood carbohydrates and lipids, and water and electrolyte balance were studied in six highly trained male marathon runners prior to and after running a marathon (26.2 miles; 42.2 km) and on control days corresponding to the above times. Fluid intake was not sufficient to maintain body weight, the runners losing approximately 2.8 kg. Estimated plasma volume losses (161 ml, 4.4%) indicated that most of the fluid loss was extravascular. Tre rose an average 2.4 degrees C and a significant negative correlation between running time and rise in Tre was observed. Glucose, fatty acids, glycerol, hemoglobin, and plasma proteins were significantly elevated after the race. Small but statistically significant increments in lactate and pyruvate were also observed. Alterations in adrenal-sympathico function were indicated by increased levels of cortisol, epinephrine, and norepinephrine.


Leg muscle metabolism during exercise in the heat and cold.

In an effort to assess the effects of environmental heat stress on muscle metabolism during exercise, 6 men performed work in the heat (Tdb = 44 degrees C, RH = 15%) and cold (Tdb = 9 degrees C, RH = 55%). Exercise consisted of three 15-min cycling bouts at 70 to 85% VO2max, with 10-min rest between each. Muscle biopsies obtained from the vastus lateralis before and after each work bout were analyzed for glycogen and triglyceride content. Venous blood samples drawn before and after exercise were assayed for lactate, glucose, free fatty acids, hemoglobin, and hematocrit. Oxygen uptake, heart rates and rectal temperatures were all significantly higher during exercise in the heat. Blood lactate concentration was roughly twice as great during the heat experiments as that measured in the 9 degrees C environment. Muscle glycogen utilization per 60 min was significantly greater in the heat ( - 74 m moles/kg-wet muscle) as compared to the cold exercise (- 42 m moles/kg-wet muscle). On the average, muscle triglyceride declined 23% during exercise in the cold and 11% in the heat. The findings of an enhanced glycolysis during exercise in the heat is compatible with earlier studies which demonstrate a decreased availability of oxygen due to a reduction in muscle blood flow.


Blood flow in thigh muscle during bicycling exercise at varying work rates.

16 male subjects exercised at 25, 50, 75, 90, 100 and 120% of VO2max on a von Döbeln bicycle ergometer. The muscle mass was measured in a whole body counter. Muscle blood flow (MBF) estimated from the rate of 133Xe clearance from m. rectus femoris showed a levelling-off at about 0.5 1 of blood per min and liter of muscle tissue (equal to an irrigation coefficient of 0.5 min-1) at work rates above 50 to 60% of VO2 max. This concurs with clearance data from the literature. However, when MBF is calculated from VO2, muscle mass, and reliable values for a- vo2 differences, MBF in the present subjects would: 1. Not level off before 90 to 100% VO2max, 2. reach a value of 1.0 min-1. The underestimation of MBF calculated from 133Xe clearance and the levelling-off shown by this method may be due to a systematic error inherent in the method, the 133Xe clearance being diffusion limited at high flow rates.


Sensitivity to catecholamines and histamine in the trained and in the untrained human organism and sensitivity changes during digestion.

Blood pressure and heart rate responses elicited by norepinephrine, epinephrine and histamine were compared before and after duodenal acidification in 27 trained and untrained young healthy men. Before acidification trained subjects showed a lower sensitivity to norepinephrine and histamine and a slightly lower one to epinephrine. In order to explain these phenomena physical training was supposed to affect alpha, respectively beta adrenergic responsivity in a dissimilar manner. This suggestion was confirmed by the isoproterenol studies, too. Duodenal acidification decreased the catecholamine-induced pressor responses. Depressor responses to histamine did not change in the untrained subjects, whereas they increased in the trained group. Thus, during digestion the histamine-induced depressor responses of trained subjects tended to approach that of untrained people.


Methodical aspects of perceived exertion rating and its relation to pedalling rate and rotating mass.

Methodical aspects of the relationship between pedalling rate and rotating mass and perceived exertion rating (PER; Borg, 1962) were studied in trained, untrained, and ill subjects in bicycle ergometry. Pedalling rate varied between 40 and 100 rpm, work load steps were 5, 10, 15 and 20 mkp/sec in the healthy subjects, and 2.5, 5, 7.5 and 10 mkp/sec in the patients. PER decreased with increasing pedalling rate in all healthy subjects. In the patients, PER increased moderately at work load of 2.5 mkp/sec, but decreased at higher work loads up to 80 rpm, followed by a slight increase at 100 rpm. Higher mass of the flywheel, studied in 6 trained subjects, lowered the PER insignificantly. In the healthy subjects, test criteria, such as reproducibility, reliability, sensitivity, and linearity remained almost unaffected by pedalling rate. In patients, increasing pedalling speed diminished reproducibility and sensitivity. The strictness of the PER work load relationship is lowered at higher pedalling rate, especially at 100 rpm. When using the PER scale, pedalling rate has to be considered as an factor of main influence.


Cardiovascular effects of venous blood from missile wounds.

A cross-infusion model including two dogs was used. At intervals 20 ml of blood from the iliac vein of one dog (donor), was injected during 60 sec into the iliac artery in which flow was measured electromagnetically in another dog (recipient). 1 min after missile trauma of the hind leg of the donor, the withdrawn venous blood possessed strong dilator property, manifesting itself by a transient, significant increase of blood flow in the registered artery of the recipient. This dilator property of venous blood from the traumatized region was, however, only apparent following wounding by missiles of high velocity.


Secretin-induced bile secretion, bile acid output and blood supply to the liver in the dog.

Bile secretion and blood flow in the portal vein and the hepatic artery were determined in cholecystectomized anesthetized dogs before and during continuous infusion of varying doses of secretin. Secretin increases bile volume without alteration of bile acid output. Hepatic arterial flow was not altered by any dose. However, high doses of secretin increased portal venous blood flow significantly. It is concluded that secretin action on liver blood flow mirrors mainly superior mesenteric arterial vasodilation and seems to be a rather pharmacologic than a physiologic response. Bile secretion is not influenced by changes of hepatic blood flow in the physiologic range.


Renal cortical necrosis induced in rabbits by local perfusion followed by systemic administration of endotoxin.

In an experimental model adapted to avoid severe systemic lesions, renal cortical necrosis was frequently induced in rabbits by repeated administration of endotoxin. The experimental model involves perfusion of the kidney in situ with endotoxin. Another dose of endotoxin was given intravenously 24-28 h before or after the perfusion. A significant effect of the perfusion or the endotoxin administered first was reflected by the pronounced unilateral morphological alterations appearing after the second administration of endotoxin. Some of the kidneys with cortical necrosis had band- or ring-shaped areas of preserved juxtamedullary tissue. Heavy infiltration of leukocytes at the borders of these areas called for commentation of the histological picture of infarcts, incorrectly described even in most modern textbooks.


Active immunization against tetanus in man. I. Duration of anamnestic reaction after one dose of vaccine.

Fifteen men, 30-40 years old, who had received one dose of vaccine against tetanus, were given a booster dose 8-13 years later. Their serum content of tetanus antitoxin was determined (1) before the booster dose, (2) 3-4 days and (3) 8 days after the booster dose. A control group of ten men with 2-3 earlier vaccine injections was treated in the same way. All the men in the first group had an antitoxin content of 0.01 IU/ml or more, 8 days after the booster dose, while the comparative titres increased to more than 1 IU/ml in the control group. The advantage of a complete basic immunization is obvious, but the investigation shows that, even after one injection, the titre after a booster dose increased to a generally accepted protective level.


Hepatic arterial and portal venous flows during hemorrhage.

Changes of total liver blood flow (TLBF), portal venous and hepatic arterial flows have been investigated using a hemorrhagic model in dogs. The dogs were bled stepwise from normal blood pressure to a pressure of 50-60 mm Hg. TLBF was measured by the xenon clearance method. Hepatic arterial blood flow was measured by electromagnetic flowmeter. Portal venous flow was calculated by subtracting hepatic arterial blood flow from TLBF. Other parameters studied during the experiment were systemic arterial blood pressure and portal venous pressure. Under normotensive conditions TLBF in mean was registered as 127 ml/min X 100 g liver tissue (25 ml/min X kg body weight). The relation between the flow value in portal vein and hepatic artery was on average 2.3:1. The study shows that there was a pronounced decrease of TLBF flow during hemorrhage. Portal venous flow decreased almost parallel to TLBF, while hepatic arterial flow decreased to a lesser extent which means that there was autoregulation in this flow bed. Hepatic arterial flow successively constituted a larger part of TLBF, during hemorrhage sometimes 65% compared to the normal value of about 30%. During the study there was an increase of vascular resistance in the portal venous system and decreased resistance in the hepatic arterial bed.


Standardized induction of myocardial ischaemia in the dog.

Ischaemia in the entire left ventricular wall was induced in dogs by installment of ameroid constrictors on the left anterior descending (LAD) and left circumflex (LC) branches of the left coronary artery (series A). Ischaemia in the anterior left ventricular wall was induced by installment of an ameroid constrictor on LAD and ligation of all (series B) or part of (series C) the subepicardial intercoronary anastomoses between LC and LAD. The effect of the operation was studied by electrocardiography and coronary angiography and, post mortem by radiography of the coronary arteries and histopathology of myocardium and coronary arteries. The high early mortality rate in series A and B precludes the further use of these procedures. In series C the mortality rate was low and in half of the cases lasting myocardial ischaemia of the anterior wall of the left ventricle was produced. The development of a small myocardial infarction was inevitable, but it was healed by the time the sought after state of stable myocardial ischaemia was reached.


Vasocystostomy, a new technique of sperm output measurement.

A vasocystostomy in the rat is described, a new technique simplifying the common methods of spermatozoa output measuring. In two series of different rat strains a mean output of 21.1 X 10(6) and 29.0 X 10(6) spermatozoa per 24 h were found. Perspectives of the new method are discussed.


[Cellular distribution of elongation factor EF1 in wheat germ].

Cellular distribution of elongation factors (EF1) from imbibed then redessicated wheat embryos is determined after purification and analytical gel electrophoresis of soluble and ribosome-bound factors. Two heavy forms (EF1 H, mol. wt, 250 000) are found in cytosol while ribosome-bound factors contain a light form (EF1L, mol. wt, 45 000) with the greatest activity and a heavy form (mol. wt, 160 000) which might well be an intermediary in the recycling of ribosomal factor EF1L to soluble factor EF1H.


Genetic differences in the induction of aryl hydrocarbon hydroxylase and benzo(a)pyrene carcinogenesis in C3H/He and DBA/2 strains of mice.

The effects of 3-methylcholanthrene and 5,6- and 7,8-benzoflavones on aryl hydrocarbon (benzo[a]pyrene) hydroxylase in the liver, small intestine, lung, and skin tissues from C3H/He and DBA/2 strains of mice under the controlled lighting conditions were examined. Apparent differences were observed in the inducibility of the hepatic enzyme by the inducers between the two strains of mice, showing that the enzyme in the C3H/He mice is inducible but not in the DBA/2 mice. Comparison of the results on the enzyme induction by 3-methylcholanthrene in the liver among the parent, intercrossed, and backcrossed mice suggested that the inducibility may be inherited as a single autosomal dominant trait. However, different genetic responses to 3-methylcholanthrene and benzoflavones in the enzyme of small intestine might be considered, because a discrepancy in the inducibility of the enzyme between the liver and the small intestine from the identical mice was demonstrated when the progeny of F2 and (DBA/2 times F1) backcross were used. On the other hand, no apparent difference was found in the inducibility of the enzyme in the lung and skin between the C3H/He and the DBA/2 mice. It is assumed that the genetic regulation of the induction of aryl hydrocarbon hydroxylase was separately controlled in the respective tissues from the identical mice. By the repeated topical applications of benzo[a]pyrene, the incidence of skin cancer was higher in the DBA/2 mice than in the C3H/He mice. The relationship between the induction of aryl hydrocarbon hydroxylase and carcinogenicity in the skin is briefly discussed.


Histogenesis and growing patterns of lung tumors induced by potassium 1-methyl-1,4-dihydro-7-(2-(5-nitrofuryl)vinyl)-4-oxo-1,8-naphthyridine-3-carboxylate in ICR mice.

The oral administration of 0.01% potassium 1-methyl-1,4-dihydro-7-(2-(5-nitrofuryl)vinyl)-4-oxo-1,8-naphthyridine-3-carboxylate (NFN) in ICR/JCL mice resulted in the development of tumors in the lung, forestomach, thymus, and other areas. Special attention was directed to the incidence and growing processes of the pulmonary tumors, which usually exhibited multicentric occurrence. Adenomatous hyperplasia and adenoma were observed in 22.2% of group A (15 approximately 30 weeks), 81.3% of group B (30 approximately 38 weeks), and 91.2% of group C (38 approximately 54 weeks). One case of carcinoma in group B and 5 cases in group C were also found. Most of adenomatous hyperplasia and adenomas developed around 30 weeks after the beginning of administration, whereas the carcinomas developed after 36 weeks. The incidence of tumors was much earlier in females than in males. Further, the tumors were observed to have originated from type B alveolar epithelial cells. The growth of type B alveolar epithelial cells was first noted as an adenomatous hyperplasia along the original alveolar walls, disintegrating and virtually replacing type A alveolar epithelial cells. Accordingly adenoma noduli was formed by these proliferated type B alveolar epithelial cells. Atypical, basophilic cell clusters were found to be present within the adenomas. It was suggested that carcinomas might have originated from these atypical type B alveolar epithelial cells.


Stimulation of HeLa-S3 cell aggregation by sugars related to cell membrane in rotation culture.

Effect of various sugars related to cell membrane on the aggregation of HeLa-S3 cells was examined in a rotation culture by adding each sugar to the culture medium. Some specific sugars among those tested induced (1) greater size of aggregates and (2) higher synthesis of hyaluronic acid than that of the control. Since the addition of hyaluronidase inhibited the aggregate formation, both in the test and control cultures, with HeLa-S3 cells of both groups forming only small aggregates, it was presumed that the hyaluronic acid synthesized might be of cardinal importance for the formation of aggregates. The specific sugars producing the effect mentioned above were N-acetyl-D-glucosamine, bis(N-acetyl)chitobiose, N-acetyl-D-galactosamine, alpha-methyl-D-glucopyranoside, and N-acetyl-D-mannosamine, while D-glucosamine, beta-methyl-D-glucopyranoside, L-fucose, D-glucose, and a commercially available hyaluronic acid were ineffective. The findings obtained in the present study interestingly ran parallel with the results of previously reported study on the induction of specific biological phenomena by the cell membrane-related sugars.


Alpha1-acid glycoprotein as a hepatocyte-specific mitosis-inhibiting protein in regenerating rat liver.

The humoral inhibitor, which may play a primary role in the regulatory mechanism of cell division in the regenerating rat liver, was investigated by isolating the active principle from the ethanol-precipitate (P-3 fraction) of normal rat plasma by isoelectric focusing, and determining the exact nature of the active principle by physicochemical analysis, and studying its localization in the liver by immunofluorescence method. The active principle was demonstrated to be alpha 1-acid glycoprotein by physicochemical analysis and by the immunohistological examination to be synthesized in the cytoplasm of the hepatocyte and excreted from it before its mitosis. These results show that the humoral inhibitor or the hepatocyte-specific mitosis-inhibiting protein is alpha 1-acid glycoprotein itself. Based on these findings, a new hypothesis concerning the regulatory mechanism of cell division in general is proposed.


Two cases of mélanose neurocutanée with development of malignant melanoma: a microspectrophotometric and electron microscopic study.

Two cases of neurocutaneous melanosis with development of malignant melanoma in the Japanese are presented. The first case was a 4-year-old boy in whom a retroperitoneal melanoma appeared with giant nevi, and cerebral and spinal melanosis. The second case was a 39-year-old man, in whom a primary leptomeningeal melanoma developed with leptomeningeal melanosis and smaller pigmented nevi. Microspectrophotometric and electron microscopic studies were made on the neoplastic and non-neoplastic melanotic tissues to elucidate the histogenesis of this rare disorder. Two different patterns of nuclear DNA histograms, corresponding to melanosis and melanoma, were obtained by microspectrophotometry, Considerable variation in the ultrastructure of the melanocytes was seen by electron microscopy.


Urinary excretion of steroids by Japanese women with breast cancer.

The urinary excretion of steroids was determined in premenopausal patients with advanced breast cancer in the non-menstrual phase and in control women in the follicular phase, ovulatory stage, and luteal phase. The estrone level in the urine of cancer patients was higher than that in the urine of control women at any phase. Excretion of 17 beta-estradiol in the follicular and luteal phases was similar in the urine of control women and cancer patients, but estradiol and estriol levels in the ovulatory stage of control women were higher than those of cancer patients. The estriol level in the urine of control women in the luteal phase was also higher than that of cancer patients. Total 17-ketosteroid was lower in the urine of cancer patients than in that of controls at any phase. Among the fractions of 17-ketosteroids, only dehydroepiandrosterone was excreted at a higher level by cancer patients than by control subjects.


Production of respiratory tract tumors in hamsters by benzo(a)pyrene.

Respiratory tumor induction was tested for the tumorigenic activity of benzo[a]pyrene alone. A group of 32 male and 28 female Syrian golden hamsters were given weekly intratracheal instillation of 1 mg of benzo[a]pyrene suspended in isotonic saline, for 30 weeks. Squamous cell carcinoma, adenocarcinoma, anaplastic carcinoma, adenoma, papilloma, and polyps were induced in the respiratory tract of hamsters. Tumor incidences were 42.3% in males and 57.7% in females. Respiratory carcinomas were inducible in hamsters by simple instillation of a low dose of benzo[a]pyrene without using a surface-active agent or carrier-dust. These findings may be useful as a standard data for cocarcinogenesis studies when using other modality combined with benzo[a]pyrene in experimental studies.


Ameloblastic odontoma in rats induced by N-butylnitrosourea.

Four cases of ameloblastic odontomas were found in the Long-Evans rats treated with multiple oral administration of a heavy dose of N-butylnitrosourea. The tumor was found as an expansive enlargement over the mandible and was roentgen-opaque. The tumor was histologically characterized by various elements such as odontogenic epithelium, stellated reticulum tissue, enamel, and/or dentine, and was classified as ameloblastic odontoma.


Induction of ovarian tumor in rats with N-butylnitrosourea.

Considerable number (8.2 approximately 10.7%) of ovarian tumors were produced in Sprague-Dawley adult rats treated with multiple oral or intramuscular administration of heavy doses of N-butylnitrosourea. Histologically the tumor was a clear-cell adenma which reminded of embryonal testicular tissue. Electron micrograph revealed secretory function of the tumor cells. It was not possible to find histogenesis in the normal ovarian components.


Effect of cytochalasin-B on the transport of 5-fluorouracil in cultured mammalian cells.

Cytochalasin-B was found to inhibit the transport of uridine, but not leucine, in cultured fibroblastic mammalian cells. The inhibition of cellular protein synthesis by 5-fluorouracil is also lessened by the addition of Cytochalasin-B to the culture.


Hepatic tumors of rabbits induced by cycad extract.

Fifteen rabbits were administered cycad extract by gastric intubation, 1 ml/rabbit, once a week for 27 or 33 weeks. The extract contained 16.6 mg of cycasin/ml. Out of 9 rabbits surviving over 200 days, 7 animals had malignant neoplasms developing in the liver. Histological and electron microscopic examinations of the developed tumors revealed that they were malignant hemangioendotheliomas. The animals showed no proliferation of hepatic cells or any tumor development in other organs.


The influence of Cortisone on the hepatic uptake of Triton WR-1339 in adrenalectomized rats.

The effect of cortisone acetate on the hepatic uptake of Triton WR-1339 was studied in adrenalectomized rats. The hormone was found to retard the uptake of Triton in the liver, and at the same time reduce the plasma clearance of this compound. The inhibitory effect of the hormone on endocytosis was seen in purified preparations of both Kupffer cells and parenchymal cells. Triton renders the liver lysosomes progressively lighter. The change in equilibrium density (in sucrose gradients) was found to occur more rapidly in hormone-treated animals. The possibility that cortisone reduces the number and increases the size of the lysosomes is discussed. Our data indicate that the uptake of Triton in non-parenchymal (Kupffer) cells represents about 20% of the total hepatic uptake.


Steroid metabolism in Bombyx mori, I. Catabolism of ponasterone A and ecdysterone in Bombyx mori.

Ponasterone A, when injected into silkworm larvae (Bombyx mori), was hydroxylated at C-25 and C-26 with the formation of ecdysterone and inokosterone, followed by fission at C-20:C-22 to form poststerone. The catabolism of ecdysterone to poststerone was also clarified. These conversions are thought to constitute part of the detoxication mechanism for exogenous ecdysones in the insect.


The lysis effect of bull spermatozoa on gelatin substrate film methodical investigations.

Proteolytic activity in the acrosomes of ejaculated bull spermatozoa was demonstrated using an autoradiographic film as a gelatin substrate. Incubation of the spermgelatin adducts at +37 degrees C and 94% humidity, which was kept constant by ventilating an incubator with water-saturated compressed air, yielded reproducible results. Gelatin depolymerisation started adjacent to the posterior segment of the acrosome within 30 to 60 s after application of individual spermatozoa to the substrate membrane and, finally, increased to a white circular digestion area enveloping the entire sperm head. The observed gelatinolysis seems to be mainly caused by acrosin, the trypsin-like acrosomal proteinase. This conclusion is supported by the positive correlation (r = +0.83, P is less than or equal to 0.01) found between the mean values of the lysis areas of individual spermatozoa on gelatin films and the acrosin activity of the sperm population measured with Bz-Arg-OEt as substrate after acidic extraction of the spermatozoa. In addition, prior saturation of the substrate layers with acrosin inhibitor (SSPI-I, II) from boar seminal plasma prevented the lysis reaction. Extraction of acrosin from the spermatozoa before application to the gelatin membranes resulted in a complete loss of any proteolytic activity. If spermatozoa were stored for 4 to 6 days at +4 degrees C or -20 degrees C in Tris buffer and afterwards applied to the substrate layer, lysis areas of individual spermatozoa differed markedly. Spermatozoa from undiluted ejaculated frozen at -20 degrees C showed no proteolytic effect on gelatin films. In general, there was a high correlation (r = +0.83, P is less than or equal 0.01) between the number of "living cells" characterized by live-dead staining and the percentage of spermatozoa active on the substrate membranes.


[Enzyme induction in Streptomyces hydrogenans. IV. (1) Qualitative and quantitative changes in RNA content and RNA synthesis during induction].

For isolation of RNA, freeze-dried cells of Streptomyces hydrogenans were disrupted by grinding with kieselguhr. Application of diethylpyrocarbonate (diethyl oxydiformate) in an extraction procedure yielded undegraded nucleic acids of high purity. Best separation of extracted nucleic acids was achieved by electrophoresis on 2% mixed agarose-acrylamide gels. After application of 11beta, 21-dihydroxy-4,14 (20)-pregnadien-3-one to the culture medium, the amount of acid-precipitable RNA in the cells decreased to 50% within 20 min. Concimitantly, the rate of incorporation of precursors into RNA is much slower immediately after addition of the inducer but increases during the next 2 h. 3-4 h after induction there is no difference in the RNA content of induced and control cells. Degradation of stable as well as unstable RNA was observed. Simultaneous addition of inducer and rifamycin inhibits the production of 20beta-hydroxysteroid dehydrogenase, suggesting the synthesis of special mRNA for this enzyme. Based on experiments with antibiotics, the half-life of total mRNA was calculated to be 3 min in control cells, and about 4 min in induced cells. Using a double isotope labelling technique, we established the existence of specific mRNA in the induced cells. Together with the longer half-life of mRNA in the induced cells, the increased transcription may allow the 40-fold stimulation of the synthesis of 20beta-hydroxysteroid dehydrogenase.


The cell growth-promoting factor.

It was recently shown that macromolecular serum proteins [1-3] as well as some of their hydrolyzed products, especially peptides of molecular weight around 5000[4] and even much less[5,6], are able to promote the growth of cells. This paper describes how the serum proteins were separated by salt precipitation and polyacrylamide electrophoresis into various albumin and globulin fractions and their growth-promoting activities ascertained. Subsequently, these macromolecules were treated with alkali, acids or proteolytic enzymes, and the activity of the products obtained was determined. We also isolated growth-promoting peptides from the liver by enzymatic hydrolysis, followed by gel filtration, or by ultrafiltration through Diaflo membranes.


Sterically hindered disulfide bridges in cystine diketopiperazine, cysteinyl-cysteine disulfide and derivatives.

L-Cystine diketopiperazine (1), L-cysteinyl-cysteine disulfide -HCl (2), L-cysteinyl-cysteine disulfide methyl ester -HCl (3), and t-butyloxycarbonyl-L-cysteinyl-cysteine disulfide methyl ester (4) are investigated by CD, ultraviolet, 13C NMR, infrared and laser Raman spectroscopy. The temperature dependence of the 13C NMR signals of 1 reveals an exceptionally high energy barrier of deltaGNo. = 15.8 +/- 0.2 kcal/mol for the reversible change in helicity of the inherently dissymmetric disulfide bridge of 1. The P-helical diastereomer predominates in dimethyl-sulfoxide at 25 degrees C, with 80-85% of the molecules having this configuration. The Cotton effects of 1 are larger and show smaller temperature coefficients than the conformationally more mobile cystine compounds 2 and 3. After dissolving crystals of 1 in 95% ethanol there is a time-dependent decrease of the ellipticity of the negative Cotton effect at 225 nm, indicating a conformational change in going from crystal to solution. Besides 1, 2 and 3 are at present the only known examples of cystine derivatives with C-S-S-C torsional angles around 90 degrees, which do not exhibit optical activity in the long wavelength disulfide absorption, as is predicted for 1 from the Linderberg-Michl model. At 305 nm a new weak Cotton effect was discovered for 1. The solvent dependence of the CD spectra is discussed and the infrared and Raman spectra are assigned.


The influence of insulin on glucose and fatty acid metabolism in the isolated perfused rat hind quarter.

Glucose and fatty acid metabolism of resting skeletal muscle were studied by perfusion of the isolated rat hind leg with a hemoglobin-free medium. Tissue integrity was demonstrated by normal ATP, ADP and creatine phosphate levels, by a sufficient oxygen supply, and by a normal appearance of perfused muscle specimens under the electron microscope. The rates of glucose and fatty acid uptake, and of lactate, alanine, glycerol and fatty acid release were constant over a perfusion period of 60 min. Insulin (1 unit/l) caused a more than threefold increase in glucose uptake, a stimulation of lactate production, and a 20% increase in the muscular glycogen levels. Fatty acids and alanine release were significantly diminished by insulin, but glycerol release did not change. The uptake of oleate by the rat hind leg was dependent on the medium concentration in a range of 0.7-1.9mM oleate, and was stimulated by insulin. Glucose uptake was not influenced by oleate, whether sodium was present or not. When the leg was perfused with [1-14C]oleate, 75% of the incorporated fatty acids were found in muscle lipids, 10% were oxidized to CO2, and 5% were recovered in bone lipids. The absolute amount of oleate oxidation was not altered by insulin. In all experiments with and without glucose in the medium, 70-80% of the 14C label incorporated into muscle lipids was found in the triglyceride fraction. In the presence of glucose, insulin significantly increased the incorporation of [1-14C]oleate into muscle triglycerides, whereas no insulin effect, either on fatty acid uptake or on triglyceride formation, could be observed when glucose was omitted from the perfusate.  The present results indicate that a "glucose-fatty acid cycle" as found in rat heart muscle does not operate in resting peripheral skeletal muscle tissue. They also demonstrate that the stimulating effect of insulin on muscular fatty acid uptake and triglyceride synthesis is dependent on glucose supply. This finding can be intrepreted as a stimulation of fatty acid esterification by sn-glycerol 3-phosphate derived from an increased glucose turnover, which is in turn due to insulin.


[The chemistry of a chitin-protein from crayfish (Astacus fluviatilis) (author's transl)].

A chitin-protein complex is obtained from crayfish (Astacus fluviatilis) by gentle decalcification with acetic acid and EDTA. The complex is treated with lithium rhodanide, urea, anhydrous formic acid, pronase, papain, anhydrous formamide or 1N NaOH. The first three of these substances have little or no effect on the stability of the chitin-protein complex. The enzymes remove most of the protein, and the last two reagents remove all of it. The protein remaining bound to the polysaccharide after treatment of the chitin-protein complex with pronase or papain is relatively rich in glycine. Quantitative analysis yielded values for the acetyl, glucosaminyl and amino acid residues which reproduce the composition of the corresponding chitin-protein complex. In these calculations however, allowance must be made for the fact that glucosamine is partly destroyed by the acid by hydrolysis and interferes with the determination of basic amino acids. The results of the present work suggest that the chitin in crayfish is present in the form of a stable complex with protein, possibly held together by covalent binding of the protein to the chitin, with glycine as the connecting amino acid.


[Repair replication in synchronous Ehrlich ascites cells after reversal of a methotrexate block (author's transl)].

If synchronous Ehrlich ascites cells are arrested at the beginning of the S phase by a methotrexate block, and the block is reversed by addition of thymidine to the culture, the cells incorporate a considerable amount of this thymidine into parental DNA within the first minute. These newly synthesized segments are irregularly distributed among the total parental DNA. This kind of distribution is expected if these segments are the result of excision repair. On the basis of the kinetics of the process this repair probably is DNA synthesis in regions already excised during the methotrexate block.


[The secondary structure and nitrocellulose affinity of freshly replicated DNA from Ehrlich ascites cells (author's transl)].

Hydroxyapatite chromatography and isopycnic Cs2SO4 centrifugation normally yield no indications of single-stranded DNA when that fraction of replicating DNA from Ehrlich ascites cells which can be separated by nitrocellulose chromatography is analyzed. Single-stranded DNA is detected by both methods if the DNA is fragmented by ultrasound before the nitrocellulose chromatography. The digestion of this DNA fraction by single-strand-specific nucliase leads to a loss of its binding to nitrocellulose and of the indications of single-stranded DNA. The loss for the affinity to nitrocellulose is also observed when the corresponding fraction separated from unfragmented DNA is digested by endonuclease. It is suggested that replicating DNA is bound to nitrocellulose by means of single-stranded gaps on the replication fork. These gaps are apparently too small to be detected within large, otherwise entirely double-stranded molecules by hydroxyapatite chromatography and Cs2SO4 centrifugation. In the case of nitrocellulose-binding ultrasound fragments, this relation seems to be more favorable because of the separation of most of the residual double-stranded part. It is demonstrated that sonication of helical DNA also generates a small amount of fragments with some single-stranded character. The effects observed with replicating DNA could be distinguished from these artifacts.


[Purification of the acetylcholinesterase from human erythrocytes by affinity chromatography (author's transl)].

The acetylcholinesterase from human erythrocytes was released from the plasma membrane with 0.2% Triton X-100 at low ionic strength and purified by two affinity chromatography steps on Sepharose-bound m-[6-(6-amino-caproylamino)caproylamino]phenyltrimethyl-ammonium. The synthesis of the inhibitor is described. The purified, detergent-free acetylcholinesterase was obtained with a specific activity of 4270 U/mg (158000-fold purification) and a 28% yield. The enzyme is a glycoprotein and aggregates in the absence of Triton X-100 into higher molecular complexes. The molecular weight was estimated by sodium dodecylsulfate electrophoresis to be 80000 +/- 3000 in the presence of 2-mercapto-ethanol and 154000 +/- 6000 in its absence.


Studies on deoxyribonucleoproteins. IX. Subfractions of deoxyribonucleoproteins from rat and mouse liver.

By carrier-free continuous electrophoresis, deoxyribonucleoprotein from rat and mouse liver could be separated into two subfractions. The more anodic fraction (DNP I), comprising 5 - 8 per cent of the total, contains fewer proteins (two types of histones only). [3H]Cyclophosphamide caused in vivo a 2.5 times higher alkylation of the DNA in in DNP I than of the DNA in DNP II. These and additional results led to the suggestion of a structural model with DNP I as a spacer in the deoxyribonucleoprotein fiber.


The effect of alpha2-macroglobulin from bovine serum on bovine alpha-chymotrypsin.

Bovine alpha2-globulin contains a protein which increases the activity of bovine alpha-chymotrypsin against synthetic substrates. The active protein fraction migrates slowly on polyacrylamide gel electrophoresis, so it was named slow alpha2-globulin (Salpha2). The fraction was isolated from bovine serum and purified. Its sedimentation constant S20 was 18.5 S. It was thus identified with the alpha2-macroglobulin (alpha2M). By kinetic studies, the dissociation constant of the alpha-chymotrypsin-alpha2 M complex was calculated to be of the order of 10(-7) l/mol. The purified alpha2 M was shown to bind alpha-chymotrypsin at a definite rate. If the binding ratio was assumed to be 1:2, the molecular weight was calculated to be about 8 X 10(5).


[Basic peptides from bee venom, I: isolation, reduction and reoxidation of apamin and MCD-peptide (author's transl)].

A simple isolation of melittin, apamin and MCD-peptide from bee venom is described. The peptides were separated using Sephadex G-50 and Biogel P-4 followed by preparative column chromatography using silicagel. Apamin and MCD-peptide were converted to the tetra-S-sulfonate derivatives by oxidative sulfitolysis. After reduction of these derivatives by mercaptoethanol followed by cautious air-reoxidation, the disulfide bridges can be reconstituted. The biological activities of the reoxidation products are 50 to 60% for apamin and 70 to 80% for MCD-peptide.


Sphingolipid metabolism in Bacteroideaceae.

The lipid composition of the anaerobic Bacteroides thetaiotaomikron has been analyzed. Sphingomyelin, ceramide phosphinicoethanolamine, free even-numbered and branched chain sphingosine bases and ceramide represented about 50% of the total lipid extract. The main ester phospholipid was phosphatidylethanolamine. The alkali-stable sphingophospholipids were predominantly N-acylated with 3-hydroxypalmitic acid, whereas the ester phospholipids are preferentially substituted with normal even and odd-numbered and branched-chain fatty acids. When Bacteroides was grown in a medium supplemented with labelled palmitic acid, this fatty acid was utilized for acylation reactions and to a large extent for the de novo synthesis of sphinganine. This long-chain base was incorporated into the sphingolipids and was also present in free form. The 3-hydroxypalmitic acid present in sphingolipids is not derived from palmitic acid, since labelled palmitate did not serve as a precursor. Free sphinganine added to the culture medium was also utilized efficiently for the biosynthesis of the sphingolipids by growing Bacteroides cultures. The 3H/14C ratio in sphingomyelin and ceramide phosphinicoethanolamine is the same, when [1-14C]palmitic acid and [3-3H]sphinganine serve as precursors. Sphingomyelin, which is usually only present in higher animals, is synthesized de novo in this Bacteroides strain.


Superoxide dismutase activity of Cu(Tyr)2 and Cu, Co-erythrocuprein.

Crystalline Cu(Tyr)2 and homogeneous Cu2Co2-erythrocuprein were prepared. The reactivity of each chelated Cu2 compound with superoxide was studied by pulse radiolysis at pH 7.6 +/- 0.1 and compared with the reactivity of native erythrocuprein (superoxide dismutase). Superoxide anions were generated by a 40-ns pulse of 1.81-MeV electrons. The yield of O2 ranged between 6 - 60 muM. The kinetics of the spontaneous O2 decay were second order; in the presence of Cu2 complexes the reaction was first order with respect to O2. Taking into account the effect of the different Cu2 concentrations on the O2 decay, second-order rate constants for the reaction of chelated Cu2 with O2 were obtained. For an equivalent of Cu2 in either erythrocuprein or Cu, Co-erythrocuprein, a numerical value of 1.3 +/- 0.1 x 10(9) M-1S-1 was calculated. Surprisingly, the same value was obtained employing Cu(Tyr)2. The highest rate constant was measured for the hydrated Cu2 (2.7 x 10(9) M-1S-1). In the presence of a biologically significant chelating agent such as serum albumin, a marked decrease in the Cu2aq-induced superoxide dismutation was observed. This was not the case when the dismutation in the presence of either the Cu2 of native erythrocuprein or Cu, Co-erythrocuprein, or those Cu2 ions chelated with tyrosine or certain di- and tripeptides was measured.


Superoxide dismutase micro assay in biological material.

A micro assay for the rapid and convenient determination of superoxide dismutase activity in limited amounts of biological material was devised and successfully employed. The combination of the formazan derivative colour formation induced by reaction of O2theta with nitroblue tetrazolium and a suitable analytical polyacrylamide gel electrophoresis system was used. It was possible to show that the reactivity of soluble superoxide dismutases in polyacrylamide gels was proportional to the enzyme concentrations employed. Bovine erythrocyte Cu, Zn-superoxide dismutase (EC 1.15.1.1) (erythrocuprein) served as a standard throughout. To measure the degree of superoxide dismutase activity, a gel-scanning apparatus was usedThe integrated scanning curve of the unstained portions of the gel proved linearly proportional to the logarithm of the superoxide dismutase activity in the range between 10(-12) and 7 X 10(-11) mol of the bovine enzyme. Although the absolute integral is dependent on the different staining conditions, the slope of the superoxide dismutase calibration curve is highly reproducible. Superoxide dismutase added to crude liver and brain homogenates could be fully detected using this assay. Thus, biological material including nucleic acids, enzymes, lipids etc. do not inhibit this reaction.


[Investigations on peptide synthesis with polymeric supports: synthesis and characterization of the deamido-calcitonin-M-(19-32)-tetradecapeptide by use of the soluble support polyethyleneglycol and coupling of the products to the insoluble polystyryl-benzhydrylamine resin (author's transl)].

The synthesis of the deamido-calcitonin-M-(19-32)-tetradecapeptide Boc-Phe-His-Thr(Bzl)-Phe-Pro-Gln-Thr(Bzl)-Ala-Ile-Gly-Val-Gly-Ala-Pro-OH (I) by the use of the soluble polymeric support polyethyleneglycol is described. For product characterization the following compounds were prepared:H-Phe-His-Thr(Bzl)-Phe-Pro-Gln-Thr(Bzl)-Ala-Ile-Gly-Val-Gly-Ala-Pro-OH and H-Phe-His-Thr-Phe-Pro-Gln-Thr-Ala-Ile-Gly-Ala-Pro-OH. The analyses of these products by ion exchange chromatography indicates that the correct tetradecapeptide sequence was synthesized to more than 90%. Despite high yields in the coupling reactions, crude I was inhomogenous due to partial cleavage to O-benzyl ether bonds. In order to use I for the total synthesis of calcitonin M, it was coupled to the insoluble polystyryl-benzhydrylamine resin. 0.08 mmol of peptide was coupled per g of resin. Treatment of this peptide resin with hydrogen fluoride yielded H-Phe-His-Thr-Phe-Gln-Thr-Ala-Ile-Gly-Val-Gly-Ala-Pro-NH2.


[Effect of glucocorticoides on the release of amino acids in the perfused rat hindquarter (author's transl)].

The release of amino acids by skeletal muscle was studied in the isolated perfused rat hindquarter. Adrenalectomy depressed the formation of glutamine and alanine as well as the efflux of all other amino acids measured. Betamethasone--a synthetic glucocorticoid--caused a significant increase in the efflux of nearly all amino acids up to the level of normal controls. The release of amino acids was also increased in perfused hindquarters of diabetic rats. On the other hand, insulin exhibited a depressing effect on the release of amino acids by hindquarters of normal rats. The metabolic integrity of the muscle tissue was proved by measuring creatine phosphate, ATP, ADP and water content as well as by the significant insulin effect on glucose uptake and on [14C]leucine incorporation into muscle proteins.


Effects of arginine homologous and other guanidino compounds on the ATP level and glucose oxidation in isolated fat cells.

The arginine homologues 2-amino-3-guanidinopropionic acid, 2-amino-4-guanidino-butyric acid and 2-amino-6-guanidinocaproic acid (= homoarginine) were synthesized and transformed into their methyl esters. The latter, together with arginine methyl esters. The latter, together with arginine methyl ester, arginine diethylamide and some guanidino compounds without the arginyl structure (agmatine, isopentyl-guanidine and n-butylbiguanide) were examined with regard to their behaviour on isolated fat cells, concerning the adrenalin-induced depression of the ATP level and the stimulation of glucose oxidation. The homoarginyl and arginyl derivatives counteracted the effect of adrenalin by re-elevating the ATP level, and thus they exerted an insulin-like activity. The esters were slightly active, whereas the arginine diethylamide and agmatine had a marked effect. The shorter homologues of arginine were totally inactive. However isopentyl-guanidine and butylbiguanide followed the effect of adrenalin: they additionally lowered the ATP level and therefore they acted in opposition to insulin. For comparative reasons the same compounds were tested with regard to their effects on glucose oxidation. The results were consistent with those quoted above: the homoarginyl and arginyl derivatives (agmatine included) forced the glucose oxidation similarly to insulin, the shorter homologues were inactive, isopentylguanidine and butylbiguanide decreased it.


[Syntheses of oligopeptides related to the insulin sequence B 22-25 (Arg-Gly-Phe-Phe) (author's transl)].

Syntheses of peptides with the sequences Gly-Phe, Gly-Phe-Phe, Arg-Gly-Phe and Arg-Gly-Phe-Phe are described. They were performed with the free acids, methyl esters and caramides. The peptides correspond partially or directly to the insulin sequence B 22 - 25 (Arg-Gly-Phe-Phe), the tetrapeptide amide or tetrapeptide methyl ester of which shows insulin-like activity (l.c.[1,2]). For testing the structural specificity of the arginyl residue, the following peptides were also synthesised: NG-NO2-Arg-Gly-Phe-Phe-NH2 and -OMe, Orn-Gly-Phe-Phe-NH2 and Cit-Gly-Phe-Phe--NH2. In connection with the above, the syntheses of the new derivatives Nalpha,Ndelta-Z2-L-ornithine p-nitrophenyl ester and N-Boc-L-citrulline p-nitrophenyl ester are described. All peptides were synthesised conventionally.


[Biological activity in vitro and in vivo of different insulin derivatives (author's transl)].

The biological activities of des-octapeptide-insulin, des-AsnA21-des-AlaB30-insulin, des-GlyA1-des-PheB1-insulin and trimethionyl-insulin were studied in vitro and in vivo. In vitro we measured, using the isolated diaphragm of the rat, the disappearance rate of glucose in the incubation medium, the incorporation of glucose into the glycogen of the diaphragm and the apntilipolytic activity in the isolated fat cell model. The incorporation of [U-14C]glucose after intraperitoneal injection into the glycogen of the diaphragm and the fat pad tissue was studied in vivo, as well as the incorporation of 14C from [U-14C]glucose into the lipids of the fat pad tissue of the rat.


[Biosynthesis of glutathione. VI. Ocurrence and level of gamma-glutamylcysteine in human red cells (author's transl)].

The concentration of the glutathione precursor gamma-glutamylcysteine in hemolysates of human red cells was determined using three different methods. a) The [14C]N-ethylmaleimide derivatives of the intracellular sulfhydryl compounds were separated on a cation exchange column. The amount of [14C]N-ethylmaleimide glutamycysteine was radiochemically determined. b) With purified glutathione synthetase, an enzymatic endpoint metabolite determination was performed with [14C]glycine as one substrate. The amount of labelled glutathione formed from the gamma-glutamylcysteine present was measured radiochemically.


5-Oxoprolinase from rat kidney, I. Assay, purification, and determination of kinetic parameters.

A new assay for the determination of 5-oxoprolinase activity is described. The enzyme 5-oxoprolinase was purified from rat kidney 285-fold to apparent homogeneity, as judged by analytical disc electrophoresis and discontinous polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate. The specific activity of the preparation was 122 mU/mg of protein. A complete initial rate kinetic analysis of the forward reaction catalyzed by 5-oxoprolinase was carried out using 5-oxo-L-proline and MgATP2theta as substrates. The computer-fitted double reciprocal plots showed intersecting patterns indicating a sequential mechanism. The data were fitted by weighted linear regression analysis using the complete equation for bisubstrate reactions. The limiting Michaelis constants for 5-oxoproline and MgATP2theta were calculated to be 31.6 +/- 2.3 muM and 172.7 +/- 11.5muM, respectively. The maximum forward rate is 1.2 +/- 0.02 mumol X min-1; the turnover number 7.0 min-1.


5-Oxoprolinase from rat kidney, II. Molecular weight determinations.

The molecular weight of 5-oxoprolinase from rat kidney was estimated by gel filtration on Sephadex G-200 and G-150 to be 460 000 +/- 30 000. A value of 230 000 +/- 10 000 was obtained by zonal sedimentation in a sucrose gradient. Polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate yielded a molecular weightof 115 000 +/- 6 000. It is concluded that 5-oxoprolinase consists of four subunits of 115 000 daltons each. The dissociation or aggregation behavior of the enzyme seems to be influenced neither by the presence of the substrates 5-oxo-L-proline and MgATP2theta nor by the presence of the stabilizing compounds glutathione mercaptoethanol or dithioerythritol.


Transpeptidation in sequence analysis. Investigations concerning a native and a synthetic hexapeptide.

On sequencing a hemoglobin a peptic hexapeptide with the amino acid composition Asp2, Gly, Val, Lys2 was isolated. Cleavage of this peptide with Tos-Phe-CH2Cl-trypsin resulted in the fragments Asp-Lys, Gly-Asp, Val-Lys, Asp--Lys-Gly-Asp, Asp-Lys-Val-Lys and Val-Lys-Gly--Asp. From these data two possible but inconsistent sequences for the hexapeptide can be derived: Asp-Lys-Val-Lys-Gly-Asp and Val-Lys-Asp-Lys-Gly-Asp. Fragments obtained by other cleavage procedures, direct sequencing of the globin peptide-chain with a sequenator as well as X-ray data of this hemoglobin show, that only the sequence starting with Asp occurs in the intact protein. Therefore the peptide Asp-Lys-Gly-Asp must have been formed by transpeptidation during sequence work. In order to verify this, Asp-Lys-Val-Lys-Gly-Asp was synthesized by an unequivocal, conventional procedure. Tryptic digestion of this hexapeptide also resulted in ASP-Lys-Gly-Asp in addition to the expected fragments. Thus it has been shown for the first time, that sequencing conditions may alter the constitution of a peptide.


[Insulin analogues with permuted A chain N-terminus (author's transl)].

By partial synthesis insulin analogues were prepared in which the amino acid in position 1 of the A chain was permuted. Glycine in position A 1 was exchanged for leucine, tert.- butyloxycarbonylvaline, valine, proline, lysine as well as glutamic acid. Two pathways of partial synthesis were followed: Firstly, des-1-glycine-A-chain S-sulfonate was reacted with active esters of tert.-butyloxycarbonylamino acids. The ensuing modified A-chains were combined with natural B-chain to give A1-permuted insulins. In the second procedure, the preparation of tris-Boc-[A1-leucine]insulin was accomplished by reaction of Boc-leucine N-hydroxysuccinimide ester with NalphaB1,NepsilonB29-bis(tert.-butyloxycarbonyl)-des-A1-glycine-insulin. The protected insulin derivative had been prepared by combination of des-glycine-A-chain with Nalpha1,Nepsilon29-bis(tert.-butyloxycarbonyl)-B-chain. The deprotected analogues differed considerably in their CD-spectra from insulin and possessed low in vitro biological activities of 2.5-17%. Crystallization attempts failed. Thus, the introduction of side chains in position A1 distorts the conformation sterically and decreases the biological activity.


Studies on polypeptides. V. Improved synthesis of human proinsulin C-peptide and its benzyloxycarbonyl derivative. Circular dichroism and immunological studies of human C-peptide.

An improved synthesis of human C-peptide is described. Five fragments: 33-39, 40-46, 47-49, 50-54 and 55-63 were used in the total synthesis. In the fully protected C-peptide the N-terminal alpha-amino function was blocked by a benzyloxycarbonyl group and the carboxyl and serine hydroxyl functions were blocked by t-butyl protection.  The latter protecting groups were removed by trifluoroacetic acid to obtain N-alpha-benzyloxycarbonyl human C-peptide which, on catalytic hydrogenation, yielded human C-peptide. The immunoreactivity of the prepared human C-peptide was tested and found to deviate slightly from the human C-peptide synthesized earlier by another route. When tested in the immunoassay, human pancreatic extracts containing natural C-peptide (or fragments thereof) showed dilution patterns identical to that of the new synthetic C-peptide but different from that of the previously synthesized batch of C-peptide. The possible explantation for the observed differences in the immunoreactivity is discussed.


Ward environment in an inpatient drug dependence treatment unit: II. Attempts to improve ward environment.

Ward activity and atmosphere in a Drug Dependence Treatment Unit were found to be poor in a previous study. The results of two attempts to improve ward environment are reported. The arrival of an occupational therapist had a very slight and restricted effect on the level of patients' constructive activity, and produced only one significant change on the Ward Atmosphere Scale; patients perceived an increase in Practicality. There was, however, a consistent improvement in patients' Vigor and Friendliness mood scores following the arrival of the occupational therapist. A pilot project using cigarettes as reinforces for patient constructive activity produced little sustained effect. Possible reasons for the difficulties in managing drug-dependent patients are discussed, and suggestions made for improvement.


Estimating heroin use in an urban area.

The purpose of this pilot project was to develop a verifiable method of empirically estimating heroin usage in a defined geographic area. If such a methodology could be constructed, appropriate officials could employ the procedure to promote planning, prevention, and education projects, and to evaluate on-going programs. Thus, should such a technique be developed, it could replace more traditional and empirically unconfirmed methods. Employing data furnished by local law-enforcement social, and medical agencies, a questionnaire was developed, pretested, and applied to a selected sample of the stipulated population.


Mood, primary heroin withdrawal, and acute methadone administration.

This study was designed to: describe the internal state of addicts undergoing primary withdrawal, determine how the internal state is altered by the alleviation of withdrawal stress through an acute dose of methadone, and to determine a predictor of the probability of a patient completing detoxification. Sixty-seven male and female heroin addicts accepted for methadone detoxification were administered a 65-item Profile of Mood States (POMS). Forty-nine of the Ss were tested immediately prior to receiving their initial dose of methadone and again 45 minutes later, while the remaining 18 Ss were only tested 45 minutes after receiving their methadone in order to control for the effects of repeated testing. Each of the subscales measured by the POMS showed a significant decrease in mood disturbance following methadone administration (p less than .0001); these differences could not be attributed to the effects of repeated testing. Factor analysis of pre- and post-methadone treatment scores indicated a two-factor structure prior to receiving methadone and a single unified mood state after the alleviation of primary withdrawal stress. The Anger subscale, administered immediately prior to receiving methadone, was a highly significant predictor of the completion of detoxification (p less than .005). Further implications of the results were discussed.


A clinical tool with heroin addicts: the urine stall.

Detection of illicit drug use by means of urine analysis has become a standard procedure in the treatment of drug addicts. Withholding urine is a way in which an addict can compromise the effectiveness of urine analysis. This behavior can be a form of acting out and should not be interpreted in a superficial manner. An attempt should be made to explore the latent psychological material it may represent.


Potential dangers of cannabis.

Cannabis is not a harmless drug. The potential dangers of cannabis are briefly reviewed in this report. The above-mentioned observations on cannabis users should be kept in mind and carefully examined by all physicians. One could expect that as more potent cannabis preparations become available, some of the toxic manifestations which now seem rare might become more frequent. Some of the remarks about the dangers of cannabis may not be proved in future studies, and they may represent only our anxiety. However, prior to the elimination of these fears, no steps should be taken toward the legalizing of marijuana. At present there is no scientific evidence that cannabis is less harmful than either tobacco or alcohol. The opposite may be true. The analogy can be drawn between opium and cannabis. The permissive attitude toward the use of opium can easily lead to the use of morphine and other opiates. If we legalize the use of marijuana, we cannot prevent the use of more dangerous derivatives of cannabis; namely, hashish, cannabis oil and THC, itself. In my opinion, in the light of our present knowledge, legalizing of marijuana could be hazardous both for the individual and for society.


Women workers in the health service industry.

The health service industry is unusual in that most of the skilled as well as unskilled workers are women, although the industry is largely controlled by men. Women are hired because they constitute an inexpensive, available, and seemingly powerless work force. Women enter health service because they have few alternatives to the low-paying, dead-end jobs found there. Health service occupations are organized like craft unions, with rigid hierarchical separations and control by the top occupation. Conflicts between men and women-between management and workers-are often played out as conflicts between occupations. Challenges to physicians come from various nursing specialties as well as from technical professions. Physicians in turn create lower-level occupations which challenge the nurses' status. Increasing industrialization alters the pattern of conflict, creating opportunities for individual bureaucratic mobility as well as favorable conditions for unionization drives. Unionism is often held back by sex, race, and professional conflicts, which must be overcome if the status of women is to be changed in the industry.


Vaginal cancer: an iatrogenic disease?

Presently we are witnessing two unique occurrences in the field of public health: the first demonstration of transplacental carcinogenesis in humans and the first drug-induced cancer epidemic in women under age 30. This article examines the current status of the vaginal cancer epidemic and possible reasons for the failure of governmental health agencies to recall and test the generation of females who were exposed to diethylstilbestrol (DES) in utero. Epidemiologic evidence indicates that the large majority of "DES daughters" may develop adenosis. The carcinogenicity of other estrogens in wide use is examined. It is pointed out that, although vaginal cancer in daughters exposed to DES in utero provided the clinical evidence to secure a Food and Drug Administration ban on DES as an additive to cattle feed, the FDA approved a new use of DES as a "morning-after pill" contraceptive even though the contraceptive contains 833,000 times the amount of DES banned for human consumption in beef. The lack of standards of informed consent in the testing of the morning-after pill on university women and the additional risk this presents to DES daughters are discussed. The sociopolitical and economic contributing factors to the vaginal cancer epidemic and the extent to which the scientific direction of medical care is influenced by economic factors are examined. Public health measures which might prevent the occurrence of such man-made epidemics in the future are recommended.


The origin of (Kolmer's) epiplexus cells. A combined histomorphological and histochemical study.

The morphology and the histochemically proofed enzyme pattern of the (Kolmer's) epiplexus cells found at the telencephalic chorioid plexus of humans indicate that these cells are of monocytogenetic origin, that they are monocytes and monocytogenetic macrophages respectively.


The development of microperoxisomes in the cells of the proximal tubules of the kidney and epithelium of the small intestine during the embryonic development and postnatal period.

Microperoxisomes were identified in the cells of proximal tubules of the kidney and small intestine of mice from 13-19 days old embryos, in suckling period and adult animals. Examination of both tissues after incubation in diaminobenzidine medium revealed anucleoid microperoxisomes closely related to the granular endoplasmic reticulum and occasionally found in close relation to it. The number of microperoxisomes within the cells considerably increases with the advanced stage of differentiation of both studies tissues. In the cells of proximal tubules the changes in shape and size of microperoxisomes are also observed with a higher stage of differentiation. These studies indicate that the formation and augmentation of microperoxisomes passes strictly parallelly with the differentiation, i.c. with the ability of cells to perform their specific functions.


Cholinergic innervation of the gastric wall of the cat.

The inbuilt intrinsic cholinergic nervous apparatus of the gastric wall of the cat was studied by using two thiocholine methods for mapping the acetylcholinesterase-positive nerves and nerve cells. A rich distribution of acetylcholinesterase-positive nerves was observed in all layers of the gastric wall, except the superficial half of the lamina propria (with the epithelium), which was completely devoid of acetylcholinesterase activity, and the submucosa, in which a scarce distribution of large nerve fascicles and nerve trunks was observed. Acetylcholinesterase-positive ganglia were observed both in the subserous layer and in the myenteric plexus of Auerbach, whereas none were recognized in the submucous plexus of Meissner. This obviously fits well to the results of some electrophysiological experiments which indicate that the submucous plexus of Meissner includes an important intramural pathway from the extrinsic vagus nerves to the antrum region; so the submucous plexus of Meissner seems to be mainly involved in direct rapid conduction of nerve impulses without integrative activities, like a cable. Certain clear differences exist in the pattern of organization of the cholinergic intrinsic nervous apparatus within the different layers of the gastric wall in the fundic and pyloric regions. These differences seem to correspond quite logically to the different types of motor, secretory and neurohumoral activities of these main regions of the stomach. The activity of the non-specific cholinesterases was localized both in the neural elements and the smooth muscle, as well as in some epithelial cells.


Glutamine transport and metabolism by mitochondria from dog renal cortex. General properties and response to acidosis and alkalosis.

Mitochondria from dog renal cortex were incubated with L-[14Cglutamine. Glutamate metabolism was prevented by inhibitors so that glutamate accumulated either in the mitochondrial matrix space or in the medium. The formation and accumuation of glutamate formed from glutamine and the distribution of glutamine in the mitochondrial fluid spaces were studied. In the matrix space glutamate rapidly reaches levels over 5 times that of glutamine in the medium. A more gradual accumulation occurs in the medium as glutamate is transported out of the mitochondria. Addition of an energy source such as succinate to the medium accelerates glutamate formation. A Km of 0.6 mM appears to govern the reaction at low concentrations of glutamine; at about 4 mM an abrupt change kinetics occurs with a Km of 5 mM above that level. Both NH4+ and glutamate inhibit glutamine metabolism and phosphate stimulates it, but little effect glutamate or phosphate occurs at low levels of these substances. The pH optimum of the reaction is between 7.4 and 7.8. Mersalyl and p-chloromercuribenzoate strongly inhibit glutamate formation; N-ethylmaleimide and bromcresol green have weaker inhibitory actions, and borate increases the reaction rate. In the presence of mersalyl, glutamine is striclly confined to the outer space of mitochondria and none is detectable in the matrix space. Similarly at ) degrees glutamine is confined to the simultaneously determined sucrose or mannitol spaces...


Bronchoprovocation: effect on priming and desensitization phenomenon in the lung.

Priming, or increased sensitivity to antigen, has not been demonstrated in the lung and could play a role in asthmatic symptomatology during seasonal pollen exposure. It is also an important consideration in the design of any experimental protocol requiring serial bronchoprovocations with antigen. Thirteen patients with a history of asthma symptoms during the pollen season and a positive skin test to ragweed extract were selected. Patients were given bronchial challenge out of season on 4 successive days with stepwise inhalations of antigen, and airways conductance was monitored in the body plethysmograph. Antigen dose-response curves were drawn, and the cumulative dose required for a 35% reduction in specific airway conductance was calculated and designated Provocation Dose (PD35). No regular trend toward either priming or desensitization was noted. The daily changes in antigen sensitivity did not correlate with daily variation of baseline pulmonary function. To determine if there was any priming due to natural exposure to pollen, 9 patients were brought back and rechallenged during the pollen season with no significant increase in bronchial sensitivity to ragweed extract. The PD35 method provides figures useful for comparing dose-response curves and shows a one-log variation from day to day. Any evaluation by bronchial challenge of antigen sensitivity or drug efficacy must take into account such variation.


The pharmacokinetics of dihydroxypropyltheophylline: a basis for rational therapy.

Dihydroxypropyltheophylline (dyphylline) was administered in a single dose on 7 different days to 7 normal subjects; 5 mg/kg was given by intramuscular injection and oral tablet; 10 mg/kg was given by these routes and also in 3 different liquid formulations. Dyphylline is rapidly absorbed, and bioavailability is independent of route of administration or formulation. The pharmacokinetics of dyphylline do not appear to be dose-dependent in the dosage range studied. The mean half-life of dyphylline is 2.11 +/- 0.36 hr; because of its short half-life and unproved efficacy, current dosage recommendations for dyphylline must be revised greatly. Further efficacy trials utilizing the pharmacokinetic data from this study are essential.


Detection of IgG antibody to deoxyribonucleic acid by the radioallergosorbent test.

IgG antibodies to heat-denatured calf thymus deoxyribonucleic acid (DNA) single-stranded DNA:ss-DNA) in the sera of patients with systemic lupus erythematosus (SLE) could be detected by means of solid-phase radioimmunoassay. Single-stranded DNA coupled covalently to microcrystalline cellulose and 125I-labeled anti-human gamma chain were used for this test. The principle of this test is known as the radioallergosorbent test (RAST). This paper describes the usefulness of the RAST for the detection of human IgG antibody. Specificity of this test was examined by an inhibition test using ss-DNA and double-stranded DNA (ds-DNA). A trial for the detection of IgM and IgA antibodies to ss-DNA was also attempted.


Biphasic local reaction to a new insulin (U-100).

The phenomenon of localized insulin reaction is described and illustrated by the report of a case. The patient's unusual response consisted of an "early" and a "late" phase to single-peak and single-component insulins. No definite immunologic mechanism was established on the basis of immunologic studies or from histologic examination of skin biopsies.


A mathematical model of aging processes. III. Comments on the relative importance of serum cholesterol and blood pressure levels on mortality.

The distributions of serum cholesterol, blood pressure, and their values added together, were examined on a cross-sectional basis for a Montreal population. The results showed that all three distributions seem to be represented better by a log normal than by a normal distribution; but in each case, there was no pronounced increase in the skew with age. The distribution of the systolic blood pressure values exhibited an increasing variance with age, whereas the distribution of the serum cholesterol values remained approximately similar with age. The distribution of the two parameters added together also did not alter significantly with age. These results are consistent with the assumption that, if the serum cholesterol value of an individual changes, the individual will be placed at a substantially greater risk than if changes occur for blood pressure levels. Subpopulations of particular concern, with respect to changes in serum cholesterol levels, are those which undergo weight changes and those which smoke cigarettes.


Protective effects of atromid-S in vivo on mitochondrial stability in tissues of aged hamsters.

Atromid-S and nicotinic acid (hypocholesterolemic agents) were tested in vivo for their effects on mitochondrial yields and oxidative phosphorylation in tissues of aging hamsters. It was observed that atromid-S increased mitochondrial yields from both heart and skeletal muscle (39 and 64%, respectively), while nicotinic acid increased the yield only from skeletal muscle (97%). Efficiency (P:0 ratios) and rates of oxidative phosphorylation remained unchanged by drug treatment. Atromid-S treatment only partially protected against loss of controlled phosphorylation on prolonged storage of heart and skeletal muscle mitochondria of aging hamsters. It appeared that atromid-S may produce its effects on mitochondrial yields by decreasing the fragility of the mitochondria from the aging animals. The effects of the drug were not permanent, and values returned to control levels after 3 weeks from cessation of treatment.


The pathogenesis of hypophyseal fibrosis in aging: its relationship to tissue iron deposition.

An investigation was made of 101 hypophyses from male and female human subjects dying from unrelated diseases. Careful microscopic examination revealed that fibrous replacement of the gland begins after the fourth decade. There is a gradual increase in iron deposition in this connective tissue after the 4th decade. It is suggested that the fibrous replacement of the hypophysis is a consequence of repeated bouts of passive congestion with diapedesis of red cells through the walls of anoxic capillaries. This pattern is akin to that of brown induration of the lung. In this manner, the active cells of the hypophysis may gradually be replaced by collagenous tissue. Because of the inherent slowness of this process, the gland seems to adapt quite well to a reduction in functional elements, thus circumventing significant clinical effects.


Human growth hormone and cortisol response to insulin stimulation in aging.

The influence of age on plasma growth hormone (HGH) and cortisol response to i.v. insulin (0.1 U/kg of body weight) was evaluated in 32 healthy subjects whose ages ranged between 20 and 84 years. A significant reduction in HGH response to insulin was observed with aging. In the young (20-34 years), middel-aged (35-49 years), and elderly (53-84 years) groups, average HGH peaks were 46.51 +/- 7.37, 29.95 +/- 5.35, and 14.31 +/- 2.39 ng/ml while average HGH areas were 2.911 +/- 0.484, 1.654 +/- 0.316, and 0.699 +/- 0.149 mug-min, respectively. Since insulin's hypoglycemic effect became less rapid with aging, this could, in part, explain the progressive decline in the HGH response to insulin. This phenomen may also be attributed to histological changes occurring in the pituitary with aging. Moreover, cortisol response was similar to all three age groups. These findings suggest that, while HGH response to insulin is correlated with age, adrenal response does not show any important modifications with aging.


Acute drug reactions among the elderly.

Research in the field of social gerontology has not examined rug use and abuse amont the aged despite the potential significance of this issue to many students of gerontology. This report, based on an analysis of the records of patients who were treated for acute drug reactions at Jackson Memorial Hospital in Miami, Florida, during 1972, examines characteristics of acute drug reactions among a sample of 60 elderly patients and compares these characteristics with those of other age cohorts. The findings show that acute drug reactions were more likely to occur among whites and females, that a substantial number of the reactions followed the ingestion of two or more substances, that roughly one-third of the admissions were directly related to suicide attempts, and that the vast majority of acute drug reactions among the elderly admissions involved the ingestion of legally manufactured and distributed drugs.


Dementia in the elderly--a search for treatable illnesses.

Forty patients with senile dementia were evaluated prospectively with laboratory and radiologic investigations, including computerized axial transverse tomography. Five patients were believed to have potentially treatable illnesses causing the dementia. After treatment, the dementia in one patient with hypothyroidism was completely resolved. One patient with pernicious anemia was markedly improved, and a second patient with hypothyroidism was somewhat improved. One patient with a brain tumor was not treated, and a severely demented patient with possible normal pressure hydrocephalus died shortly after shunting without improvement. The discovery of these unsuspected illnesses and the gratifying response to treatment in some suggests the importance of systematic evaluation in all patients with senile dementia.


Response of institutionalized geriatric mental patients to a twelve-week program of regular physical activity.

Twenty-three geriatric patients from a mental institution were randomly assigned to three groups-a social group, an exercise group, and a control group. Subjects participated in the study for 12 weeks. Heart rate at rest, exercise and recovery, balancing ability, total daily activity level (diary method), and a self-care inventory were recorded for analysis. Data were collected at the start, the 8th and 12th weeks of the study, and 4 weeks after the termination of treatment. A Groups X Sex X Measurement Period factorial design, using multivariate analysis of variance tested the effect of the three factors. The trials effect was significant, as were several univariate tests for the sex effect and group by sex interaction. Data were discussed in terms of programmed physical activity for geriatrics, its value, and the limitations of evaluating such behavior in geriatric populations.