Affinity chromatography of lipoxygenases.

A number of aminohexyl agarose derivatives of unsaturated fatty acids have been prepared and evaluated as materials for the affinity chromatography of soybean and pea lipoxygenases. A practical method for a one-stage purification of soybean lipoxygenase-1, with a purification factor of 16, is described, using either linolenate or docosa-4,7,10,13,16,19-hexaenoate as ligands. Results show that alleged competitive inhibitors do not cause sharp elution from the affinity column, and that there is an increasing specificity of binding and sharpness of elution as the proportion of unsaturation in the ligand is increased. These results are discussed in terms of the relative importance of the types of bonding involved in enzyme-substrate binding.


Assembly of Bacillus subtilis phage phi29. 1. Mutants in the cistrons coding for the structural proteins.

The effect of mutations in the cistrons coding for the phage structural proteins has been studied by analyzing the phage-related structures accumulated after restrictive infection. Infection with susmutants in cistron 8, lacking both the major head and the fiber protein, does not produce any phage-related structure, suggesting a single route for the assembly of phage phi29; infection with ts mutants in this cistron produces isometric particles. Mutants is cistron 9, coding for the tail protein, TP1, produce DNA-free prolate heads with an internal core; these particles are abortive and contain the head proteins HPO, HP1 and HP3, the upper collar protein NP2 and the nonstructural proteins p7, p15 and p16. Mutants in cistron 10, coding for the upper collar protein, NP2, produce DNA-free isometric heads also with an internal core; they contain the head proteins and the nonstructural protein p7, suggesting that this protein forms the internal core. Mutants in cistrons 11 and 12, coding for the lower collar protein, NP3, and the neck appendages, NP1, respectively, give rise to the formation of DNA-containing normal capsids and DNA-free prolate particles, more rounded at the corners than the normal capsids and with an internal core; the DNA-containing 11-particles are formed by the head proteins and the upper collar protein; the DNA-free 11-particles contain, besides these proteins, the nonstructural protein p7 and a small amount of proteins p15 and 16. The DNA-containing 12-particles have all the normal phage structural proteins except the neck appendages, formed by protein NP1; the DNA-free particles are similar to the DNA-free 11-particles. After restricitive infection mutant sus14(1241) has a delayed lysis phenotype and produces a phage burst higher than normal, after artificial lysis. It produces DNA-containing particles, identical to wild-type phage, which have all the normal phage structural proteins, and DNA-free prolate particles, more rounded at the corners than the final phage particles and with an internal core; the last particles contain the same proteins as the DNA-free 11 or 12-particles. These particles could represent a prohead state, ready for DNA encapsulation. None of the DNA-containing particles have the nonstructural proteins p7, p15 or p16, suggesting that these proteins are released from the proheads upon DNA encapsulation.


Assembly of Bacillus subtilis phage phe29. 2. Mutants in the cistrons coding for the non-structural proteins.

The effect on phage morphogenesis of sus mutations in the cistrons coding for nonstructural proteins has been studied. Mutants in three cistrons analyzed that are involved in phage DNA synthesis, as well as in cistron 16 which codes for a late nonstructural protein, produce prolate capsids which are more rounded at the corners than complete phage heads and have an internal core; they contain the head proteins, the upper collar protein and protein p7, not present in mature phage particles. Mutants in cistron 7 do not produce capsids nor other phage-related structures; this result and the presence of p7 in phage capsids suggest an essential role in capsid assembly for this protein. The protein product of cistron 13 is probably needed for a stable DNA encapsulation since mutants in this cistron produce mainly DNA-free complete phage particles and only about 10% of uninfective DNA-containing complete phage. Cistron 15 codes for a late, partially dispensable, nonstructural protein which is present in the DNA-free capsids produced after infection with the delayed-lysis mutant sus14(1242), used as the wild-type control, or with mutants in cistrons 9, 11,12 and 13. Proteins p15 and p16 are probably involved in the encapsulation of viral DNA in a prohead.


Eye findings in the 13 trisomy syndrome.

The gross and microscopic eye findings in the first historic case of the 13-trisomy syndrome included: severe microphthalmia, coloboma of the ciliary body, cataracts, detached retina, and retinal dysplasia.


Characteristics and behaviour of arylesterase in human serum and liver.

Human arylesterase is localized in liver microsomes where the presence of different electrophoretic bands corresponding to the serum bands can be recognized. Serum arylesterase is mainly a result of liver activity and its high level might be explained by a low rate of elimination in urine. The behaviour of arylesterase towards inhibitors shows certain similarities to that of some of the proteases, such as trypsin. The clinical value of serum arylesterase determination in assessing liver function is confirmed by its isoenzyme behaviour in cirrhosis and porto-caval shunt.


The effects of digoxin and beta-methyldigoxin on the heart rate of decompensated patients with atrial fibrillation.

Eighteen patients with atrial fibrillation were given digoxin 0.13 mg twice daily for 3 weeks and beta-methyldigoxin 0.10 mg twice daily for another 3 weeks. At the end of each 3 week period an exercise test was performed and the effects on the heart rate of the two drugs were compared. No difference in heart rate was obtained at rest, whereas the heart rate after 6 min of exercise was higher during treatment with digoxin (131 beats/min) than when the patients were taking beta-methyldigoxin (124 beats/min). There were no significant differences between digoxin and beta-methyldigoxin in their effects on the ECG (R-R intervals, T-wave, Q-T duration). The plasma concentrations of the two glycosides were determined by radioimmunoassay and by 86Rb-uptake inhibition assay. Comparable plasma concentration values (1.0 ng/ml for digoxin, 1.1 ng/ml for beta-methyldigoxin, mean values) were obtained by radioimmunoassay, but the 86Rb-technique gave significantly higher values (mean 1.5 ng/ml) for beta-methyldigoxin. It is concluded that beta-methyldigoxin is equal to digoxin for producing slowing of the heart rate in patients with atrial fibrillation.


Biochemical measurements in Paget's disease of bone.

In a group of eighty-three patients with untreated Paget's disease of bone, plasma alkaline phosphatase activity (AP), plasma non-protein-bound hydroxyproline concentration (PHP) and urinary excretion rate of total hydroxyproline (THP) were closely correlated with each other but not with fasting plasma concentrations of calcium or inorganic phosphate. Probit plots of AP and THP showed log-normal distributions overlapping the normal ranges.


The influence of phentolamine, an adrenergic blocking agent, on insulin secretion during surgery.

A rapid intravenous glucose load (20 g) was given with a phentolamine infusion during and after elective abdominal surgery. Plasma levels of glucose, free fatty acids, and insulin were measured to investigate the influence of surgical stress on insulin secretion. When Ringer's lactate solution was infused into a control group of subjects during surgery, plasma levels of insulin did not change during and after the surgery while plasma levels of glucose and free fatty acids increased gradually during this period. Similar results were also noted in another control group in whom only Ringer's lactate solution and phentolamine had been infused. This evidence suggests that insulin secretion responding to endogenous hyperglycemia is suppressed during surgery. In the group which was given the glucose load during infusion of only Ringer's lactate solution, plasma levels of insulin significantly increased soon after the glucose load and the gradually decreased. In another group which was given the glucose load during infusion of Ringer's lactate and phentolamine, plasma levels of insulin also increased significantly after the glucose load and remained elevated during surgery. The maximum increment of plasma insulin after the glucose load in the latter group was significantly higher than that in the former group. From these results it is suggested that suppression of insulin secretion by surgical stress is inhibited by the alpha blocking agent phentolamine.


Insulin disappearance rate in pregnant and non-pregnant women, and in non-pregnant women given GHRIH.

The half-life of injected monocomponent insulin, as judged by radioimmunoassay, has been found to be 3-3 1/2 min in a group of healthy women. This value is unaffected by pregnancy. Tests were repeated in non-pregnant women during an infusion of GHRIH to ensure that endogenous insulin secretion was suppressed by the dose of insulin given. Similar values were obtained.


Cholesterol metabolism in Papua New Guineans.

Cholesterol metabolism has been studied in a group of healthy males drawn from a New Guinea highland population whose plasma cholesterol concentration is low by Western standards and does not rise with age. Mean values from compartmental studies in six subjects were: pool A mass 289 mg/kg; production rate 1069 mg/day or 17.4 mg kg-1 day-1. The average rate of endogenous synthesis measured by sterol balance in twelve men was 751 mg/day or 12.4 mg kg-1 day-1. There was evidence of probable suppression of synthesis by about 500 mg/day when 1 g/day of egg yolk cholesterol was added to the diet of six subjects for 5 weeks. In the latter group, mean cholesterol absorption was 39% (390 mg/day) but the plasma cholesterol concentration did not rise, possibly reflecting compensatory redution in synthesis. It is suggested that these results were unexceptional, except possibly for the failure of the plasma cholesterol concentration to rise when cholesterol was ingested, despite gross differences in diet and many other factors.


Plasma aldosterone during extracellular fluid volume expansion in patients on regular haemodialysis.

The influence of extracellular fluid volume expansion on the plasma aldosterone concentration (PAC) was investigated in five anephric and six non-nephrectomized patients on regular haemodialysis, and compared to a control group of four anephric and four non-nephrectomized patients. Plasma-renin activity, cortisol, Na+, and K+ were measured together with the PAC during the investigation. In anephric patients the PAC remained constant during the control period as well as during extracellular fluid volume expansion by infusion of 350 mmol of 20% mannitol. In the non-nephrectomized patients PAC diminished after mannitol infusion. The decline in PAC was correlated with the basal levels of PAC and the plasma renin activity. It is concluded that 5% extracellular fluid volume expansion has no direct influence on the regulation of PAC in patients without the renal renin-angiotensin system and that the regulation of PAC in anephric patients in the present investigation is probably mediated by changes in potassium and ACTH.


Regional vascular responses to asphyxia in the rabbit.

Organ blood flow was measured in eight spontaneously breathing male New Zealand white rabbits exposed to a 50 min period of asphyxia. The results were compared with eight control animals. Cardiac output and arterial pressure did not change. There was increased flow to the heart, brain and diaphragm. Flow to the kidneys and adipose tissue was reduced. Flow to the gastro-intestinal tract, liver, skeletal muscle and carcass was unchanged. Prolonged moderate asphyxia produces preferential blood supply to vital organs and maintains flow to the skeletal musculature and the gastro-intestinal tract; their blood supply is diverted mainly from the kidneys. These changes are similar to those seen in haemorrhagic and endotoxic shock in the rabbit.


Screening for medullary carcinoma of the thyroid in families with Sipple's syndrome: evaluation of new stimulation tests.

In search of new practical diagnostic methods for the early diagnosis of hereditary medullary carcinoma of the thyroid (MCT) calcitonin release has been studied following induction by pentagastrin, cholecystokinin-pancreozymin (the C-terminal octapeptide, C8-CCK, and the native swine extract), and ethanol in eighteen cases of MCT (all but one clinically occult), three 'borderline cases', seven first degree relatives of patients with hereditary MCT and thirty-five healthy controls. Pentagastrin, subcutaneous (s.c.) or intravenous (i.v.), induced a pronounced and rapid increase of serum calcitonin within 2-5 min. The elevation was roughly proportional to the tumour mass as estimated at operation. Seventeen out of eighteen MCT patients responded to s.c. pentagastrin with a significant increase in serum calcitonin and the response correlated well with that induced by calcium infusion test. Only two blood samples, at times 0 and 5 min, were necessary for diagnosis. In the MCT patients, i.v. pentagastrin produced more pronounced elevations of serum calcitonin than did s.c. pentagastrin, whereas no increase was seen in the control group. The subjective discomfort caused by i.v. pentagastrin was somewhat more intense but lasted shorter than that induced by s.c. administration. No serious complications were seen. All of nine MCT patients responded to C8-CCK with increments in serum calcitonin exceeding those of the control group and both of two responded similarly to the native cholecystokinin-pancreozymin extract. Generally the serum calcitonin response was lower and more variable after C8-CCK than after s.c. or i.v. pentagastrin, and the subjective discomfort was also more pronounced with abdominal cramps during the injection. Ethanol in the dose used was the least effective stimulator for serum calcitonin release. Clinically suspected MCT carriers with palpable tumours can be diagnosed by determination of the basal, i.e. non-stimulated serum calcitonin levels. Other possible Sipple genome carriers, who are at the time clinically healthy with normal basal serum calcitonin, should be subjected to a s.c. or i.v. pentagastrin stimulation test at each examination. These tests are much simpler to perform than a calcium infusion, test, but seem to have about the same sensitivity.


Reduced binding of added zinc in serum of patients with decompensated hepatic cirrhosis.

Binding of zinc to macromolecules when added to serum was measured by ultrafiltration. At a final ratio of 0.6 mol zinc per mol albumin, zinc dissociable by glycine was significantly greater in diluted serum from patients with decompensated alcoholic cirrhosis than in normal serum samples similarly prepared. Low serum zinc concentrations, often observed in patients with alcoholic cirrhosis, may be related in part to a reduced affinity of plasma albumin for exchangeable zinc.


Induction of uterine protein synthesis by synthetic progestins.

The capacity of artificial progestins, each possessing a C-19 methyl group, to stimulate protein synthesis in the uteri of castrated New Zealand White rabbits was studied in an attempt to screen for this type of progestational activity. It is apparent that modifications of the 4-pregnen-3,20-dione molecule not only alter total protein production, but also effect changes in the synthesis of the uterine protein, blastokinin, in comparison with the effect of unaltered progesterone. This indicates that altering the molecular configuration of progesterone offers contraceptive potential through its ability to influence the preimplantation uterine environment.


Adverse reactions and endotoxin content of polysaccharide vaccines.

Adverse reactions occurring when 1.5 million children and young adults were vaccinated with different lots of group A meningococcal and one lot of Haemophilus influenzae type b capsular polysaccharide vaccines were analyzed on the basis of records kept by local health personnel and 19,000 special questionnaires. The incidence of anaphylactic reactions was 0.8 per 100,000 injections. A pyrogen-like reaction--high fever appearing soon after injection and disappearing within six hours--was seen in 1.8% of children 3 months to 5 years old who received the meningococcal vaccine in the first vaccination project. This reaction was most common in children below three years. Its frequency varied with different vaccine lots and was found to be correlated with the endotoxin content of the vaccines estimated by either the limulus assay or by pyrogen test in rabbits. On the basis of these results we propose that capsular polysaccharide vaccines should pass the pyrogen test with a dose of one mug polysaccharide per kg of rabbit weight. With such vaccines, the incidence of high fever was reduced to 0.5% in children of the same age group.


An audio urine-glucose analyzer for blind diabetics.

At the instigation of one of the authors (J.M.)--a young man whose blindness is due to diabetic retinopathy--a urine-glucose analyzer was designed and constructed. Its purpose and function enable him to test his urine himself (free of dependence on thers for this service) and, thereby, he can then determine and administer his doses of insulin. He and other young blind diabetics may value the degree of freedom from dependence on others than the analyzer provides and feel it helps them to be able to work and care for themselves. A description of the instrument instructions for its use, and a summary of about 20 months' experience with it are submitted.


A morphologic study of intrahepatic portal-vein islet isografts.

Isologous pancreatic islets were implanted into the portal vein of rats with streptozotocin-induced diabetes. At intervals of from one to 32 days after transplantation, the intrahepatic islet grafts were examined histologically and ultrastructurally, and their vascular supply was determined by later perfusion studies. Implanted islets were found widely dispersed throughout the liver in peripheral interlobular portal venules and surrounded by vacuolated liver cells containing large stores of glycogen. The endocrine cells were structurally normal in each interval examined. By the third day after transplantation the beta cells were depleted of secretory granules in aldehyde-fuchsin preparations. Regranulation returned by the 14th day and was associated with secretory organelle hypertrophy and hyperplasia. Islet cells were found outside the portal areas in direct apposition to hepatocytes forming distinct desmosomes by the first day. While hemoperfusion of the grafts occurred from the moment of implantation into the portal venule, a dual vascular supply derived from periportal arterial and venous sources developed by the 11th day after transplantation, establishing full vascularization of the grafts. Preliminary work is presented to show that an active ingrowth of nerves in the islet graft occurs in association with the process of vascularization.


Accumulation of myocardial triglycerides ketotic diabetes; evidence for increased biosynthesis.

The accumulation of triglycerides in the myocardium of nonketotic and overtly ketotic diabetic rats was studied. There was no increase in heart triglycerides of nonketotic rats taken off insulin treatment, although the rats exhibited several indices of diabetes. In nonketotic diabetic rats untreated with insulin, myocardial triglycerides repeatedly increased and declined to control levels. In severely ketotic rats, heart triglycerides increased about threefold and did not decline with time. In order to understand the mechanism of increase in myocardial triglycerides in ketotic diabetes, the biosynthesis of triglycerides was studied with heart homogenates. The total esterification of sn-glycero-3-phosphate was unaltered, but the synthesis of diglycerides and triglycerides was increased in the myocardium of the ketotic rat. On treatment of the diabetic rats with insulin, the synthesis of di- and triglycerides in heart homogenates reverted to control values. Thus the results of the present study demonstrate that (1) a persistent increase in myocardial triglyceride content was observed only in the ketotic diabetic rat and (2) increased synthesis of triglycerides is a factor in its accumulation in the myocardium of the ketotic rat.


Recombination and response to selection in Drosophila melanogaster.

Most biologists beleive that recombination speeds response to selection for traits determined by polygenic loci. To test this hypothesis, sixteen Drosophila melanogaster populations were selected for positive phototaxis for twenty-one generations. In some populations, balancer chromosomes were used to suppress autosomal recombination, and in others the autosomes were free to recombine. Suppression of recombination had no effect on mean rate of response to selection, though it may have increased variability in the rate of response among replicate lines. Suppressed recombination lines did not shift selection response to the freely recombining X chromosomes, despite fairly large increased in X chromosome recombination. The results suggest that in populations of moderate size, sex does not accelerate short term response to selection.


Malnutrition: a poorly understood surgical risk factor in aged cardiac patients.

This feasibility report is based on the fact that malnutrition has been recognized but too little understood in connection with surgical risk. Patients with cardiac cachexia are remarkably similar to many patients with cachexia of the aged. Cachectic patients generally go through an operation well, but their condition often deteriorates slowly and they die a few days later; they behave as if they are running out of energy reserves. Malnourished people can be divided into three categories: kwashiorkorlike, marasmic, and marasmic-kwashiorkorlike. Recognition and classification of protein/calorie malnutrition into these categories directs treatment. Recognition is based on the usual physical and laboratory tests, plus triceps skinfold/arm circumference observations; leukocyte counts, with absolute and relative lymphocyte counts; serial transferrin, globulin, and albumin assessments; and, particularly, Candida and mumps skin testing to identify the anergic state. Intravenous and oral hyperalimentation can bring about conspicuous improvement in the appearance, attitude, and ability to withstand stress--including major heart surgery--of malnourished patients. However, astute clinical balance is essential, since either oral or intravenous hyperalimentation may cause renal nitrogen overload; moreover, if intravenous delivery is too rapid, congestive heart failure may be precipitated.


Amylase creatinine clearance ratio after biliary surgery.

The amylase creatinine clearance ratio (ACCR) is considered to be a more sensitive index of acute pancreatitis than the serum amylase level. Serial ACCR estimations were undertaken in 25 patients undergoing an elective cholecystectomy. Using accepted criteria, 28% of these patients developed, in the postoperative period, biochemical evidence of pancreatic gland damage, although the serum amylase level remained normal. This raised ACCR was particularly noted in patients who had undergone an exploration of the common bile duct. The ACCR would appear to be a more sensitive index of pancreatic gland disruption secondary to biliary surgery than the serum amylase level.


Intracellular proteolytic activity during sporulation of Bacillus megaterium.

Intracellular proteolytic activity increased during incubation of the sporogenic strain of Bacillus megaterium KM in a sporulation medium together with excretion of an extracellular metalloprotease. The exocellular protease activity in a constant volume of the medium reached a 100-fold value with respect to the intracellular activity. Maximal values of the activity of both the extracellular and intracellular enzyme were reached after 3-5 h of incubation. After 7 h 20-50% cells formed refractile spores. The intracellular proteolytic system hydrolyzed denatured proteins in vitro at a rate up to 150 mug mg-1 h-1 and native proteins at a rate up to 70 mug mg-1 h-1. Degradation of proteins in vivo proceeded from the beginning of transfer to the sporulation medium at a constant rate of 40 mug mg-1 h-1 and the inactivation of beta-galactosidase at a rate of 70 mug mg-1 h-1. The intracellular proteolytic activity was inhibited to 65-88% by EDTA, to 23-76% by PMSF. Proteolysis of denatured proteins was inhibited both by EDTA and PMSF more pronouncedly than proteolysis of native proteins; 50-65% of the activity were localized in protoplasts. Another strain of Bacillus megaterium (J) characterized by a high (up to 90%) and synchronous sporulation activity was found to behave in a similar way, but the rate of protein turnover in this strain was almost twice as high. The asporogenic strain of Bacillus megaterium KM synthesized the exocellular protease in the sporulation medium, but its protein turnover was found to decrease substantially after 3-4 h. The intracellular proteolytic system of the sporogenic strain J and the asporogenic strain KM were also inhibited by EDTA and PMSF.


Growth, glucose metabolism and melanin formation in biotin-deficient Aspergillus nidulans.

Biotin deficiency resulted in an increased growth rate of Aspergillus nidulans. The activities of hexokinase and aldolase were not much changed during the growth cycle, but activities of glucose-6-phosphate dehydrogenase and NADP-linked glutamate dehydrogenase increased significantly during the exponential phase. This change was remarkable during biotin deficiency. In contrast to the higher growth rate and respiration rate during biotin deficiency the activities of NAD(P)H oxidoreductases were low. An inverse relationship between the activity of tyrosinase and melanin content was observed. A role of the DOPA-DOPA-quinone system in maintaining culture growth is suggested.


When to institutionalize the elderly.

Comprehensive evaluation and follow-up of the chronically ill aged patient may show that he can stay at home with support from family and community services. Guidelines are given for choosing a nursing home when necessary.


Phosphorus deficiency and hypophosphatemia.

Low serum phosphorus levels, sometimes associated with depletion of phosphorus stores, can engender a variety of serious, often life-threatening physiologic changes. The proximate cause of this dangerous situation is usually medical intervention in such conditions as alcoholism and diabetic ketoacidosis, which can produce a shift of phosphorus within the body unless preventive measures are instituted.


Evaluation of antigen-induced buffy coat leucocyte aggregation as a simple test of allergic reactivity.

The phenomenon of antigen-induced aggregation of human buffy coat leucocytes correlates well with skin testing using purified protein derivative of tuberculin as the antigen and in this system appears to be a simple, sensitive test of cell-mediated immunity. The same phenomenon may, however, be used as an indicator of humoral immunity. In addition antigen-induced buffy coat leucocyte aggregation is suppressed by serum from patients with Hodgkin's disease and sarcoidosis. The possible uses and also the limitations of this test are discussed.


Membrane associated antigens of human malignant melanoma. III. Specificity of human sera reacting with cultured melanoma cells.

Sera from melanoma patients, healthy donors, pregnant women and patients with types of tumors other than melanoma were tested on various melanoma lines as well as on a cultured brain tumor and adult skin fibroblasts, using a microimmune adherence test. Positive reactions against all cell lines were found in serum from each group of donors. The degree of reactivity was dependent on the cell line used. Sequential absorption with AB Rh+ and pooled platelets of more than 200 donors either reduced the titer of sera or rendered a great part of the sera negative, demonstrating that antibodies against HL-A antigens and other tissue antigens were involved. The remaining positive sera were further absorbed with pooled cells from 6- to 8-week-old fetuses. This step abolished the reactivity of most sera, indicating the relatively high frequency of antibodies in males and females against fetal antigens expressed also on melanoma and other cells. In order to determine the specificity of the few remaining positive sera, absorptions with three different melanoma cell lines, a brain tumor and fibroblasts were carried out. The results showed only partial cross-reactivity between different cell lines. No evidence was obtained from this study for the existence of a common cross-reacting membrane-associated antigen on human malignant melanoma. Antigens that could be readily detected seemed mostly to be tumor-associated fetal antigens.


Condylar fractures of the mandible. I. Classification and relation to age, occlusion, and concomitant injuries of teeth and teeth-supporting structures, and fractures of the mandibular body.

In 123 individuals, 138 fractures of the mandibular condyle were classified with respect to fracture level, dislocation at the fracture level, and condylar head relation to the articular fossa. The age of the patient and the location of the most distal occlusal contact were recorded as well as teeth injuries and concomitant fractures of the mandibular body. The position of the most distal occlusal contact did not influence the dislocation of the condylar fragment. Teeth injuries such as fractures and luxation were found to be associated with condylar head or neck fractures, whereas the concomitant fractures of the mandibular body were mostly seen with the subcondylar fractures. Teeth injuries as well as concomitant fractures of the mandibular body were found more frequently in patients with bilateral than unilateral condylar fractures. Fractures of teeth in the molar and bicuspid regions were most frequent on the condylar fracture side, while the concomitant fractures of the mandibular body were located to the contralateral side. Medial angulation of the condylar fragment with lateral override at the fracture level was the typical fracture in adults, and angulation without override the characteristic fracture in growing individuals. Medical override occurred both in children and in adults and seemed to be the result of more severe trauma to the chin.


The interrelation of postoperative complaints after removal of the mandibular third molar.

Quantitative data on pain, trismus, and swelling after removal of a mandibular third molar are presented. The population of 932 patients included both extractions and surgical removals. Frequency distributions of trismus and pain are given. For all complaints the behavior of the average value as a function of time is presented and these functions are compared with each other. For all values measured on the 3rd and 4th posttreatment days, correlation and regression coefficients for the effects were calculated. It is concluded that there exists a rather strong, but time-dependent, interrelationship between the complaints.


Clinical consequences of complaints and complications after removal of the mandibular third molar.

Quantitative data on postoperative complaints and complications after removal of the mandibular third molar, especially in relation to the different methods of treatment, are scarce. Mostly an objective approach is lacking. The present study attempts to provide such data and their clinical consequences. Within 1 year, 932 mandibular third molars were removed; 430 extractions served as a control group for 502 surgical removals. The procedure was fully standardized and all measurements objectively obtained. A computer was used for statistical evaluation. It appeared from this study that the possibility of postoperative complaints and complications will be smaller if the wound is not packed, if tight suturing is not done and if reflection of the mucoperiosteum is avoided. If reflection is inevitable the oblique vertical relaxing incision is preferred. Prophylactic administration of antibiotics is rejected.


The different patterns of fractures of the facial skeleton in four European countries.

A series of 1,420 Dutch patients with facial fractures was compared with series from three other European countries. Special attention is paid to the patterns of the facial fractures and the cause of those injuries. The majority of the fractures in all four countries were caused by traffic accidents. The age of the Dutch patients at the time of accident as well as the localization of the mandibular fractures differed from the ages and localizations of the patients in the other three countries. Manifold associated injuries occurred in combination with facial fractures. The need for treatment of facial fractures in well-equipped centers is important and cooperation with other specialiies is very often necessary.


A prosthetic view on vestibuloplasty with free mucosal graft.

One hundred and twenty-four edentulous patients underwent preprosthetic surgical treatment; a mandibular and/or maxillary vestibuloplasty, some of them in combination with a deepening of the floor of the mouth or a tuberoplasty. One-three years after operation vertical dimension, central relation, retention, and function of the complete dentures were evaluated. Attention was also paid to the scar region in cheek and vestibulum, to the aspect of the mucosal graft, the changes in sensibility of chin and lips, and the changes in physiognomy. The scar position in the sulcus was investigated. The patients were questioned about difficulties in speech and tongue movement, and pain experience after the operation. They were also asked whether they regarded the new denture as an improvement and the operation as successful, and whether they would recommend the operation to others.


Blood fibrinolytic activity before and after oral surgery.

Blood smaples were collected in cold O.1 M sodium citrate from 30 healthy young persons whose mandibular impacted molars were to be removed. Blood sampling was repeated immediately after operation. Fibrinolytic activity of the euglobin fraction, precipitated at pH 5.9, was measured by the fibrin plate method. Three different fibrinogen solutions were used: 1) bovine fibrinogen, 2) bovine plasminogen-free fibrinogen, and 3) human fibrinogen. The differences between preoperative and postoperative fibrinolytic activity were evaluated. No increase of fibrinolytic activity was seen in general, indicating that oral surgery does not necessarily influence the fibrinolytic activity of the blood. Dividing the patients according to sex or to a long/short surgical procedure did not influence the activity either. However, in four patients who developed alveolitis sicca (dry socket), a significant rise of activity on all the fibrin plats was seen (P less than 0.01) when compared with the variations measured in patients with normal healing. The result suggests that a rise of blood fibrinolytic activity of a certain magnitude during oral surgery may predispose to the devlopement of alveolitis sicca. Four females who took oral contraceptives showed a higher postoperative fibrinolytic activity (P less than 0.05) than the other females. The importance of this observation is to be investigated in further studies.


Operative treatment of arthrosis deformans of the temporomandibular joint: a preliminary report.

If patients with arthrosis deformans of the temporomandibular joint are treated without success by conservative means, operative therapy may be indicated. The type of operation depends on the complaints: a)resection of the auriculotemporal nerve and superficial temporal artery; or b) the same operation in combination with condylar shaving. Twenty-four joints in 14 patients underwent a denervation. Thirty joints in 19 patients were also shaved. The results were very satisfactory.


A comparison of four commonly used local analgesics.

In a double-blind, cross-over, clinical study with nine probands, the efficiency parameters of four commonly used local analgesics were evaluated: 2% Carbocain with adrenaline 1:200,000, 3% Citanest with Octapressin 0.03 iu/mul, 3% Carbocain Dental, and 2% Xylocain with adrenaline 1:80,000. It was found that 3% Carbocain Dental (no vasoconstrictor content) demonstrated significantly shorter duration of analgesia than the other solutions in the soft tissues as well as in the dental pulp. 3% Citanest with Octapressin exhibited the longest soft tissue duration, but as far as duration of tooth analgesia was concerned there was no significant difference between 3% Carbocain Dental and 2% Carbocain with adrenaline. The longest duration of tooth analgesia was shown with 2% Xylocain with adrenaline; this solution also possessed the highest frequency of analgesia and extent of analgesia. All probands personally preferred injections with 3% Carbocain Dental as injection with this solution resulted in the least discomfort.


Immunoelectrophoretic quantitation of antithrombin III. Electroimmunodiffusion in agarose gels containing heparin.

Antithrombin III (AT-III), being an alpha2-globulin, will have an electrophoretic mobility in the presence of heparin like prealbumin in agarose gels. This phenomenon was utilized to quantitate AT-III from serum and plasma by electroimmunodiffusion (EID) for 90 min agarose gels containing 75 USP units of heparin/ml gel. The method permits a rapid quantitation of AT-III from serum, citrated plasma and EDTA plasma, and a positive correlation was observed between these values and those obtained by single radial immunodiffusion (SRI). This is in contrast to quantitation of AT-III by EID in gels containing no heparin where the values for plasma showed poor correlation with those obtained by SRI.


Xenon inhalation as an adjunct to computerized tomography of the brain: preliminary study.

The purpose of this study is to determine whether computerized tomography can distinguish between brain tissue and brain tissue containing dissolved xenon at physiologic concentrations. Xenon is an inert gas of high atomic number (54), and is highly soluble in tissue, particularly in fat. Its presence in the brain after inhalation is manifested by well known anesthetic effects. Phantom studies using xenon in equilibrium at atomospheric pressure with water, corn oil, and milk samples of varying known fat content, demonstrate that xenon is detectable in all cases with a steep linear increase in change of attenuation factor (EMI number) with increasing fat content. In the rhesus monkey xenon is readily detectable at 20% inhaled gas concentration, with linear detectable at 20% inhaled gas concentration, with linear increase of attenuation factor with increasing concentration. The possible application of our findings to the study of brain pathophysiology is discussed. Since xenon is a potent although safe anesthestic, caution in clinical application is advised.


Joint remodelling and the evolution of the human hand.

A funtional morphological study has been made of the joints of the primate hand, particular emphasis being placed upon the carpometacarpal and metacarpophalangeal joints. The presumptive evolutionary history of these joints has been charted by reference to a comparative series of mammals. It has been demonstrated that the human joints have been quite strikingly modified in a number of ways, and that these evolutionary changes may be logically correlated with the refined functional attributes of the human hand. The morphological background thus established has been applied in a preliminary study of the hand bones of various fossil hominids.


A new type of inactivation of streptomycin by E. coli.

Previously described cases of streptomycin inactivation by R-factor carrying strains of E. coli have not lead to any measurable decrease in antimicrobial potency in the bulk substrate toward the culture. In these cases each cell inactivates only a few molecules. Out of 1,800 strains of E. coli we have isolated five strains which inactivate streptomycin in large amounts giving a final concentration of the inactivation product of 0.25 mg/ml in 36 hours. Unlike all other streptomycin-resistant strains in investigated these five strains were sensitive to butyl-streptomycylamine, a streptomycin derivative acting in the same way as streptomycin. The crude inactivation product has been isolated. Inorganic phosphate is liberated by treatment with alkaline phosphatase resulting in a streptomycin-like compound without any antimicrobial activity.


Cardiac glycogen repletion after exercise: influence of synthase and glucose 6-phosphate.

Previous studies have shown that cardiac glycogen content is replenished to supranormal levels after exercise. To better understand this phenomenon glycogen synthase (GS) %I activity as well as glucose 6-phosphate (G-6-P) and glycogen (G) concentrations were measured in hearts from normal and alloxan diabetic rats killed at various time intervals before or after a treadmill run to exhaustion. In normal animals at exhaustion G was reduced but GS %I and G-6-P were elevated. During 2-8 h of recovery G was elevated, %I depressed, and G-6-P unchanged when compared to their preexercise values. By contrast, diabetic hearts that had elevated G and low %I were characterized by abnormally high G-6-P concentrations. At exhaustion in these hearts %I increased significantly in response to a decrease in G. These findings suggest that glycogen repletion occurs in normal heart as a result of the combined increases in GS %I and G-6-P levels present at the cessation of work. In both normal and diabetic hearts there exists an inverse relationship between G content and GS %I activity.


Hypoxia-induced metabolic and core temperature changes in the squirrel monkey.

To identify components of reduced core temperature (Tre) with exposure to hypoxia, oxygen uptake (VO2) and skin temperatures were measured in chair-restrained squirrel monkeys during conditions of 21% O2, 11% O2, and 11% O2 + 5% CO2. Exposure to 11% O2 resulted in a 20% decrease in VO2 and a 1.8 degrees C reduction of Tre with skin temperatures closely paralleling Tre. The addition of 5% CO2 to 11% O2 effectively blocked the decrease of both VO2 and Tre. The responses of one monkey (LD) exhibiting decreased Tre (greater than 2.0 degrees C) and 20% decrease in VO2 were compared to those of another (SD) exhibiting decreased Tre (less than 0.5 degrees C) and little change of VO2 under the same experimental conditions. Increased mean arterial blood pressure (BPa) and heart rate (HR) occurred for monkey SD, while for monkey LD BPa decreased and HR increased slightly. Arterial pressure of oxygen (PaO2) and calculated arterial saturation of oxygen (SaO2) were higher, while PaCO2 was lower for monkey SD than for monkey LD. When 5% CO2 was added to 11% O2 both animals exhibited decreased PaO2, little change of Tre, VO2, PaCO2, arterial pH, and calculated SaO2, and increased BPa and HR. These data suggest that decreased Tre resulted from reduced heat production and reduced oxygen transport was primarily responsible for reduction of heat production.


Biochemical properties of Clostridium bifermentans spores.

As previously found for spores of Bacillus species, dormant spores of Clostridium bifermentans contained essentially no adenosine triphosphate, a high level of adenosine monophosphate, a high level of 3-phosphoglyceric acid, and much transfer ribonucleic acid lacking a 3'-terminal adenosine monophosphate residue. As in spores of Bacillus species, germination of C. bifermentans spores was accompanied by utilization of the 3-phosphoglyceric acid, a large increase in the adenosine triphosphate level, and the disappearance of defective transfer ribonucleic acid. In contrast to spores of Bacillus species, dormant spores of C. bifermentans contained little free amino acid.


Macromolecular composition of spores from the filamentous cyanobacterium A nabaena cylindrica.

Spores were isolated from the filamentous cyanobacterium Anabaena cylindrica, and their deoxyribonucleic acid, ribonucleic acid, and protein compositions were determined.


Density, porosity, and structure of dried cell walls isolated from Bacillus megaterium and Saccharomyces cerevisiae.

Helium displacement and nitrogen adsorption techniques were used to determine the density and porosity, respectively, of freeze-dried cell walls isolated from Bacillus megaterium KM and Saccharomyces cerevisiae. The densities were 1.302 and 1.180 g/cm3, respectively, suggesting noncrystalline solids. The porosities were extremely small, indicating that the cell walls had collapsed and become essentially impervious upon lyophilization.


Metabolism of pyrimidine bases and nucleosides in Bacillus subtilis.

In Bacillus subtilis, uracil (Ura), uridine (Urd), and deoxyuridine (dUrd) are metabolized through pathways similar to those of enteric bacteria. Ura is probably converted to uridine 5'-monophosphate by uridine 5'-monophosphate pyrophosphorylase. More than 95% of dUrd added to cultures is converted to Ura and deoxyribose-1-phosphate. Although dUrd kinase activity is detectable in vitro, this enzyme does not seem to play an important role in the metabolism of dUrd. The metabolism of cytosine (Cyt), cytidine (Cyd), and deoxycytidine (dCyd) in B. subtilis appears to be different from that in enteric bacteria. Cytosine cannot be used by Ura-requiring mutants as pyrimidine source. dCyd is deaminated by dCyd-Cyd deaminase or phosphorylated to dCyd nucleotides by dCyd kinase. Cyd is deaminated by dCyd-Cyd deaminase of phosphorylated by Cyd kinase. This Cyd kinase activity has never been reported for B. subtilis.


Glucan common to the microcyst walls of cyst-forming bacteria.

Chemical analysis indicated that D-glucose is tha major neutral monosaccharide present in the microcysts of a range of gram-negative bacteria. Varying amounts of other neutral sugars were found. The glucose was mainly present as a glucan that could be extracted from microcysts of representative strains with alkali or mild acid treatment. The glucan could be identified as an alpha-1,3-linked polymer on the basis of (i) periodate resistance of the extracted polymer and the material present in microcysts; (ii) lectin agglutination of the microcysts; (iii) lectin precipitation of the extracted glucans; and (iv) susceptibility of the glucan either in the walls or after extraction to a specific alpha-1,3-glucanase from Aspergillus nidulans, yielding glucose as the sole hydrolysis product. The galactosamine found in microcysts of Myxococcus xanthus by other workers is clearly a component of another polymer, distinct from the glucan. The presence of an alpha 1,3-linked glucan, common to microcyst walls of various bacterial genera, probably contributes to the rigidity of the walls of these forms and, inter alia, to their resistance to ultrasonic treatment. Preliminary experiments indicate that the gulcan is discarded on germination of the microcysts rather than being broken down by specific enzymes.


Rate of ribonucleic acid chain growth in Mycobacterium tuberculosis H37Rv.

Two methods were employed to measure the rate of ribonucleic acid (RNA) chain growth in vivo in Mycobacterium tuberculosis H37Rv cultures growing in Sauton medium at 37 degrees C, with a generation time of 10 h. In the first, the bacteria were allowed to assimilate [3H]uracil or [3H]guanine into their RNA for short time periods. The RNA was then extracted and hydrolyzed with alkali, and the radioactivity in the resulting nucleotides and nucleosides was measured. The data obtained by this method allowed the calculation of the individual nucleotide step times during the growth of RNA chains, from which the average rate of RNA chain elongation was estimated to be about 4 nucleotides per s. The second method employed the antibiotic rifampin, which specifically inhibits the initiation of RNA synthesis without interfering with the elongation and completion of nascent RNA chains. Usint this method, the transcription time of the 16S, 23S, and 5S ribosomal RNA genes was estimated to be 7.6 min, which corresponds to a ribosomal RNA chain growth rate of 10 nucleotides per s.


Occurrence of facultative anoxygenic photosynthesis among filamentous and unicellular cyanobacteria.

Eleven of 21 cyanobacteria strains examined are capable of facultative anoxygenic photosynthesis, as shown by their ability to photoassimilate CO2 in the presence of Na2S, 3-(3,4-dichlorophenyl)-1,1-dimethylurea and 703-nm light. These include different cyanobacterial types (filamentous and unicellular) of different growth histories (aerobic, anaerobic, and marine and freshwater). Oscillatoria limnetica, Aphanothece halophytica (7418), and Lyngbya (7104) have different optimal concentrations of Na2S permitting CO2 photoassimilation, above which the rate decreases: 3.5, 0.7, and 0.1 mM, respectively. In A. halophytica, for each CO2 molecule photoassimilated two sulfide molecules are oxidized to elemental sulfur, which is excreted from the cells. The ecological and evolutionary significance of anoxygenic photosynthesis in the cyanobacteria is discussed.


Inhibitory protein controls the reversion of protoplasts and L forms of Bacillus subtilis to the walled state.

When the cell wall of Bacillus subtilis is removed by lysozyme and the resultant protoplasts are plated on hypertonic soft agar medium, each protoplast forms an L colony. L bodies from such L colonies again plate as L-colony-forming units (CFU). However, if protoplasts or L bodies are "conditioned" by 1 h of incubation in 0.4% casein hydrolysate medium and then incubated in 25% gelatin medium for 1 h, 60 to 100% of the formerly naked cells give rist to bacillary colonies. The present experiments largely explain the mechanism responsible for the "heritable" persistence of the wall-less state in B. subtilis. It is shown that protoplasts produce a reversion inhibitory factor (RIF) which blocks reversion when the cell concentration exceeds 5 x 105 CFU/ml. This inhibitor is nondialyzable and sensitive to trypsin, heat, and detergent. Efficient reversion at 2 x 107 CFU/ml is obtained if the protoplasts are treated with trypsin after conditioning and chloramphenicol is incorporated into the gelatin reversion medium. In the presence of 500 mug of trypsin per ml, the requirement for gelatin is sharply reduced, and reversion occurs rapidly in liquid medium containing only 10% gelatin. Trypsin also stimulates reversion in L colonies growing on soft agar. Latent RIF is activated by beta-mercaptoethanol. This reagent blocks reversion of protoplast suspensions at densities of 5 x 105 CFU/ml. Comparison of the autolytic behavior of B. subtilis and of the RIF revealed that several or the properties of the two activities coincide: both are inhibited by high concentrations of gelatin, both are activated by beta-mercaptoethanol, and both have high affinity for cell wall. Going on the assumption that RIF is autolysin, models for protoplast reversion is suggested by the finding that mutants with altered teichoic acid show altered reversion behavior.


Cell density-dependent growth of Myxococcus xanthus on casein.

When Myxococcus xanthus FB was grown on 0.2% casein it exhibited a phenomenon we call cooperative growth. That is, above 104 cells per ml, both strains that were studied exhibited increasing growth rates as a function of increasing cell numbers. Between 104 and 107 cells per ml, the mean doubling times of strains YS and TNS decreased from 15.2 to 8 h and 26 to 8.5 h, respectively. The extracellular proteinase activity of the two strains was equivalent and directly proportional to cell number. Cooperative growth was correlated with increased concentration of hydrolyzed casein in the medium, suggesting cooperative hydrolysis of casein. At low cell densities neither strain was capable of measurable growth on casein in liquid media, and we have calculated that the average concentration of hydrolyzed casein in the medium was indeed too low to support growth. At low cell densities, growth on hydrolyzed casein (Casitone) was normal and independent of cell concentration. Demonstration of cooperative growth at higher cell densities supports the suggestion that the communal behavior of myxobacteria results in more efficient feeding.


Some properties of a membrane-deoxyribonucleic acid complex isolated from Bacillus subtilis.

Membrane-deoxyribonucleic acid (DNA) complexes were isolated from Bacillus subtilis by affinity for magnesium-Sarkosyl crystals. These complexes (M-bands) contained greater than 80% of the total cellular DNA; little of the remaining portion could be recovered in a secondary isolation. Isotopic labeling of the origin of replication showed this region of the chromosome to be closely associated with the cell membrane. Interruption of protein or DNA synthesis did not result in detachment of the chromosome from the membrane. Interruption of ribonucleic acid synthesis by rifampin resulted in a decreased ability to isolate DNA in the M-band. Analysis of attachment of the chromosome to membrane during the cell and replication cycles indicated that the chromosome is not released from the membrane at any time during the cell cycle.


Developmentally induced autolysis during fruiting body formation by Myxococcus xanthus.

The developmental events during fruiting body construction by the myxobacterium M. xanthus is an orderly process characterized by several sequential stages: growth leads to aggregation leads to formation of raised, darkened mounds of cells leads to autolysis leads to myxospore induction. The temporal sequence of autolysis followed by myxospore induction is consistent with the interpretation that developmental autolysis provides essential requirements for the surviving cells to induce to myxospores. At intermediate developmental times on agar plates a fraction of the cell population is irreversibly committed to lyse; i.e., lysis continues in liquid growth medium or in magnesium-phosphate buffer. Lysis is cell concentration independent and is therefore likely to be by an autolytic mechanism. The lysis sequence can be preliminarily characterized as having an early stage during which deoxyribonucleic acid synthesis continues and a later irreversible stage during which deoxyribonucleic acid synthesis does not occur. Irreversible lysis in liquid growth medium or in magnesium-phosphate buffer is initiated on agar plates during nutrient deprivation and such lysis results in the induction of a fraction of the population to myxospores. This induction is dependent upon the concentration of lysis products, thus providing evidence that developmentally induced autolysis is required for myxospore induction.


Media dependence of commitment in Bacillus subtilis.

At some time during sporulation development, cells of Bacillus subtilis develop a commitment to continue sporulation even after addition of or dilution into a fresh nutrient. The extent of commitment was measured by the titer of spores produced at the time at which the original culture sporulated maximally. Since newly formed spores of B. subtilis soon germinate in the replenished medium, the measurement of their titer, especially of heat-resistant spores, gave low values. This problem was avoided by the germination-delaying effect of methyl anthranilate (1 mM) when added together with the fresh nutrients. In a given culture, the titer of committed cells was then independent of the method by which it was measured, i.e., by the phase-bright, octanol-resistant, or heat-resistant spore titer. The time of commitment depended on the type of nutrient added. Commitment occurred earlor casein hydrolysate. The rates at which non-metabolizable amino acid analogues or the 14C from an amino acid mixture were taken up by the cells increased toward the end of growth and later declined. This decline occurred slowly and was only weakly correlated with the commitment time of an analogous amino acid.


Isolation of acetyl esterase mutants of Bacillus subtilis 168.

Five mutants of Bacillus subtilis 168 defective in an intracellular esterase activity were identified. By polyacrylamide gel electrophoresis, four of the mutants were shown to lack esterase B activity, and the fifth lacked esterase A activity. All of the back-crossed esterase mutants were able to sporulate at wild-type frequency and produce exoprotease(s) and antibiotic(s). No difference in motility could be attributed to the esterase mutation. PBS1 transduction analysis showed all the esterase B mutations to be linked to the hisA marker.


A low molecular weight inhibitor of steroid receptor activation.

Dilution at 0 degrees of rat liver cytosol incubated with [3H]triamcinolone acetonide provoked an enhanced binding of steroid-receptor complexes to nuclei. The explanation of this phenomenon was found to be an "activation" of the complexes. Dilution acted by decreasing the concentration of a cytosol inhibitor. This reaction was irreversible at 0 degrees: once activated the complexes could not be reversed to the nonactivated state by the addition of inhibitor. The presence of hormone was necessary, since hormone-free receptor molecules could not be activated by dilution. Removal of the inhibitor did not lead to activation of all complexes: after 24 h a "plateau" was attained where 55 to 70% of the complexes were activated. The inhibitor was shown to be a low molecular weight molecule by dialysis, Sephadex G-25 chromatography, ammonium sulfate precipitation, and ultrafiltration. Thus [3H]triamcinolone acetonide-receptor complexes present in a cytosol from which the inhibitor had been removed by Sephadex G-25 chromatography became spontaneously activated at low ionic strength and at 0 degrees. The inhibitor is not a steroid (at least of usual polarity) since it cannot be extracted by methylene chloride or adsorbed by activated charcoal. It is thermostable (resists to 30 min at 100 degrees). Its removal by incubation with a cation exchange resin suggests that it may be positively charged, however it is not complexed by EDTA. This inhibitor must be distinguished from a previously described inhibitor of steroid-receptor complexes binding to nuclei. The latter compound has been shown in various systems to be responsible for an artifactual saturation of nuclear acceptor by steroid-receptor complexes. It inhibits the binding to nuclear acceptors of already activated complexes and is probably a macromolecule. It is thus different from the low molecular weight activation inhibitor described in the present paper.


Primary structure of the "hinge" region of human IgG3. Probable quadruplication of a 15-amino acid residue basic unit.

The middle part of the heavy chain of IgG3 (hinge region) which covalently links the two gamma3 chains to each other, is about 4 times larger than the same region in the three other human IgG subclasses. This is probably due to a quadruplication of a 45-nucleotide DNA segment resulting in a gamma3 hinge region which is 62 amino acid residues long and consists of an NH2-terminal 17-residue segment followed by a 15-residue segment which is identically and consecutively repeated three times. The NH2-terminal 17-residue segment shows 70% homology with the repetitive 15-residue segment and appears to be the result of a small insertion and several point mutations of the same 45-nucleotide DNA stretch. Since this unit of repetition shows 60 to 70% homology with the hinge of the other IgG subclasses, it may represent the primitive IgG hinge.


Purification and properties of microsomal UDP-glucuronosyltransferase from rat liver.

p-Nitrophenol conjugating activity associated with liver microsomal UDP-glucuronosyltransferase (EC 2.4.1.17) was purified 150- to 200-fold from cell-free homogenates. The purification scheme included solubilization with the nonionic detergent Lubrol WX, anion exchange chromatography at pH 6.0 and 7.5, and affinity chromatography with UDP-hexanolamine Sepharose 4B. The enzyme purified as a phospholipid-protein complex and was shown to consist of a single polypeptide chain of molecular weight 59,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amino acid analysis indicated approximately 531 mol of amino acids/59,000 g of enzyme and a molar ratio of nonpolar to polar residues of 1.08. During fractionation, the enzyme displayed instability with such steps as gel filtration, dialysis, or ultrafiltration of dilute samples; however, upon adsorption to ion exchange resins or storage in concentrated form, the enzyme was reasonably stable. The active lipoprotein complex showed both size and charge heterogeneity as judged by gel filtration and electrofocusing. Three forms of the enzyme resolved by isoelectric focusing had isoelectric points which averaged pH 6.68, 6.56, and 6.31. Polypeptide compositions of these electrophoretically distinct phospholipid protein complexes were indistinguishable on the basis of sodium dodecyl sulfate-polyacryl-amide gel electrophoresis, suggesting that the charge heterogeneity may be the result of differences in the phospholipid content of the lipoprotein complex.


Partial purification and properties of a ribosomal RNA maturation endonuclease from Bacillus subtilis.

Data are presented on the partial purification and properties of a 5 S ribosomal RNA maturation nuclease, termed RNase M5, from Bacillus subtillis 168. RNase M5 specifically cleaves 21 and 42 nucleotides, respectively, from the 5' and 3' termini of a 5 S rRNA precursor to yield the mature (116 nucleotides) 5 S rRNA. The cleavage is endonucleolytic with the formation of 5'-phosphoryl and 3'-hydroxyl groups. Enzyme action requires divalent cations, which may be furnished by either certain metals or by polyamines. The activity is separable into two components both of which are required for activity. It appears that the same nuclease excises the 5'- and 3'-terminal segments since preparations lose the capacity to modify the two termini with an identical first order thermal decay rate. Certain features of the rRNA precursor which may be involved in cognitive interaction with RNase M5 are discussed.


Amino acid sequence of Desulfovibrio vulgaris flavodoxin.

The complete amino acid sequence for the 148-amino acid flavodoxin from Desulfovibrio vulgaris was determined to be: H3N+-Met-Pro-Lys-Ala-Leu-Ile-Val-Tyr-Gly-Ser-Thr-Thr-Gly-Asn-Thr-Glu-Tyr-Thr-Ala-Glu-Thr-Ile-Ala-Arg-Glu-Leu-Ala-Asn-Ala-Gly-Tyr-Glu-Val-Asp-Ser-Arg-Asp-Ala-Ala-Ser-Val-Glu-Ala-Gly-Gly-Leu-Phe-Glu-Gly-Phe-Asp-Leu-Val-Leu-Leu-Gly-Cys-Ser-Thr-Trp-Gly-Asp-Asp-Ser-Ile-Glu-Leu-Gln-Asp-Asp-Phe-Ile-Pro-Leu-Phe-Asp-Ser-Leu-Glu-Glu-Thr-Gly-Ala-Gln-Gly-Arg-Lys-Val-Ala-Cys-Phe-Gly-Cys-Gly-Asp-Ser-Ser-Tyr-Glu-Tyr-Phe-Cys-Gly-Ala-Val-Asp-Ala-IleGlu-Glu-Lys-Leu-Lys-Asn-Leu-Gly-Ala-Glu-Ile-Val-Gln-Asp-Gly-Leu-Arg-Ile-Asp-Gly-Asp-Pro-Arg-Ala-Ala-Arg-Asp-Asp-Ile-Val-Gly-Try-Ala-His-Asp-Val-Arg-Gly-Ala-Ile-COO. This protein is of interest as it was the first flavoenzyme for which high resolution x-ray diffraction studies were published (Watenpaugh, K.D., Sieker, L.C., and Jensen, L.H. (1973) Proc. NAtl. Acad. Sci. U.S.A. 70, 3857-3860). Ser(10), Thr(12), Asn(14), and Thr(15) were shown to bind the phosphate of the FMN while the isoalloxazine ring is positioned between Trp(60) and Tyr(98).


Inactivation of bacterial D-amino acid transaminases by the olefinic amino acid D-vinylglycine.

D-Vinylglycine (2-amino-3-butenoate) functions as a transamination substrate and irreversible inactivator of the homogeneous pyridoxal phosphate-dependent D-amino acid transaminases from Bacillus subtilis and Bacillus sphaericus. In the absence of alpha-ketoglutarate as co-substrate, vinyl-glycine causes little if any inactivation of either enzyme; in the presence of excess alpha-ketoglutarate, both enzymes are inactivated with pseudo-first order kinetics. The limiting rate constant for inactivation of the B. sphaericus enzyme is 1.9 min-1, for the B. subilis enzyme it is 0.36 min-1. The number of catalytic events before inactivation is about 450 for the B. sphaericus enzyme and about 800 for the B. subtilis enzyme; that is, about 0.2% inactivation in each catalytic cycle for the former enzyme and 0.15% for the latter. Comparisons are made with the L-aspartate amino-transferase from pig heart which is inactivated completely in one catalytic cycle and the L-alanine aminotransferase which is not inactivated in many cycles. Comparisons are also made between the likely mode of D-transaminase inactivation produced by vinylglycine and the mode of inactivation induced by beta-chloro-D-alanine.


Quantitative analysis of the change of metabolite fluxes along the pentose phosphate and glycolytic pathways in Tetrahymena in response to carbohydrates.

A metabolic scheme of glycolysis and the pentose phosphate pathway has been constructed, assuming that the reactions occur in a single compartment. From this scheme, equations are written for a system in metabolic and isotopic steady state. These allow computation of the specific activity of every carbon atom of all the intermediates of the glycolytic and pentose phosphate pathways and consequently of the flux of carbon along each step of these pathways. A sufficiently large number of well distributed measurements of incorporation of radioactive label from different positions of several substrates into intermediates or products must be made to determine all the fluxes. This is done by choosing a set of metabolic fluxes, calculating incorporation with the aid of a computer, and then manipulating the flux rates until the computed incorporations match the data. The model is used in this paper to analyze the metabolism of the protozoan Tetrahymena pyriformis. The metabolic scheme of the model is consistent with all available information on the enzyme complement of this ciliate. Cells grown to transition phase in proteose/peptone medium were inoculated into a mixture of glucose (6 mM), fructose (6 mM), ribose (3 mM), and glycerol (3 mM) and incubated for 1 h. In each of these experiments, one of the following labeled substrates was present: [1-, 2-, 6-, or U-14C]glucose; [1- or U-14C]fructose; [1- or U-14C]ribose; [1(3)-or 2-14C]glycerol. The incorporation of label from these substrates into CO2, lipid, glycogen, and RNA was measured. In contrast to earlier studies on the metabolism of 2- and 3-carbon substrates by Tetrahymena, the rate of incorporation of label from some substrates into some products (e.g. from [1-14C]glucose into CO2) changed during the incubation. To treat these time-dependent data within the framework of the steady state model, the 1-h incubation was divided into three 20-min intervals; within each of these, the rates of incorporation were approximately constant, as required for a steady state system. Measurements of the pool sizes of glucose-6-P and fructose-6-P showed that only slow changes in pool sizes occurred after the first 5 min of incubation and indicated that the system was effectively in a metabolic and isotopic steady state throughout most of the incubation. The finding that a low concentration of cycloheximide prevented the acceleration of 14CO2 production from labeled glucose suggests a role for protein synthesis in the slow adaptation to carbohydrate addition and supports the quasi-steady state treatment of this system. The expected incorporation into each product was computed for trial sets of 1, independent flux rates. A set of flux values was found which yielded a good fit to the 29 measurements made for each interval. These flux values therefore constitute a quantitative description of temporal changes in carbon flow along the glycolytic and pentose phosphate pathways during the 1st h of adaptation to the carbohydrate mixture...


Enzymic characteristics of fat globule membranes from bovine colostrum and bovine milk.

Fat globule membranes have been isolated from bovine colostrum and bovine milk by the dispersion of the fat in sucrose solutions at 4 degrees C and fractionation by centrifugation through discontinuous sucrose gradients. The morphology and enzymic characteristics of the separated fractions were examined. Fractions comprising a large proportion of the total extracted membrane were thus obtained having high levels of the Golgi marker enzymes UDP-galactose N-acetylglucosamine beta-4-galactosyltransferase and thiamine pyrophosphatase. A membrane-derived form of the galactosyltransferase has been solubilized from fat and purified to homogeneity. This enzyme is larger in molecular weight than previously studied soluble galactosyltransferases, but resembles in size the galactosyltransferase of lactating mammary Golgi membranes. In contrast, when fat globule membranes were prepared by traditional procedures, which involved washing the fat at higher temperatures, before extraction, galactosyltransferase was not present in the membranes, having been released into supernatant fractions, When the enzyme released by this procedure was partially purified and examined by gel filtration, it was found to be of a degraded form resembling in size the soluble galactosyltransferase of milk. The release is therefore attributed to the action of proteolytic enzymes. Our observations contrast with previous biochemical studies which suggested that Golgi membranes do not contribute to the milk fat globule membrane. They are, however, consistent with electron microscope studies of the fat secretion process, which indicate that secretory vesicle membranes, derived from the Golgi apparatus, may provide a large proportion of the fat globule membrane.


Tetrahymena strives to maintain the fluidity interrelationships of all its membranes constant. Electron microscope evidence.

When cells of Tetrahymena pyriformis, strain NT-1, were chilled from their growth temperature of 39.5 degrees C to lower temperatures, the plasma membrane, outer alveolar, nuclear, outer mitochondrial, food vacuolar, and endoplasmic reticulum membranes each responded in a fashion quite characteristic of the membrane type. In most cases a distinctive rearrangement of intramembrane particles, as discerned by freeze-fracture electron microscopy, began abruptly at a definitive temperature. By comparing the freeze-fracture patterns of membranes in cells grown at 39.5, 27, and 15 degrees C, it was shown that the initial particle rearrangement in a given membrane always occurred at a fixed number of degrees below the growth temperature of the cell. Gradual chilling of a cell grown at constant temperature induced these membrane changes first in the outer alveolar membrane, then, in order of decreasing response to temperature, in the endoplasmic reticulum, outer mitochondrial membrane, nuclear envelope, and vacuolar membrane. The normally stable relationships between the physical properties of the several membrane types could in some cases be reversed, but only temporarily, by fatty acid supplementation or during the initial phases of acclimation to growth at a different temperature. The system provides a unique opportunity to study the effects of environmental change upon the physical properties of several functionally distinct but metabolically interrelated membranes within a single cell.


Effect of dexamethasone on prolactin and TSH responses to TRH and metoclopramide in man.

We studied the effects of administration of dexamethasone, 2 mg orally every 6 h, for 5 days on the thyrotropin-releasing hormone (TRH)-induced release of prolactin (PRL), thyrotropin (TSH), triiodothyronine (T3) and thyroxine (T4) in 9 normal men and on the metoclopramide-induced release of PRL in 7 normal men. Dexamethasone suppressed the baseline serum levels of PRL, TSH and T3. The administration of dexamethasone blunted the PRL and TSH response to TRH; the blunted TSH response resulted in a decreased T3 and T4 response to TRH after dexamethasone. Following dexamethasone administration, the PRL response to metoclopramide, a dopamine antagonist which acts at the hypothalamicpituitary level to stimulate PRL secretion, was blunted in 7 normal men. The data suggest that short-term administration of pharmacological doses of glucocorticoids suppress the secretion of PRL and TSH by a direct effect on the anterior pituitary gland.


Graves' disease associated with familial deficiency of thyroxine-binding globulin.

Five patients presented with symptoms of Graves' disease and a marked decrease in their thyroxine-binding globulin (TBG) capacity. While thyrotoxic, mean values +/- SD for the 5 patients were: total thyroxine (TT4) 8.8+/-2.0 mug/100 ml (normal range 4.4-9.3); TBG capacity 6.1+/-1.1 mug T4/100 ml (normal range 16-24); free thyroxine index (FTI) 25.3+/-8.9 (normal range 3.6-9.3); and total triiodo-thyronine (TT3) 244+/-56 ng/100 ml (normal range 80-160). When euthyroid, both TT4 (2.8+/-0.8 mug/100 ml) and TT3 (68+/-12 ng/100 ml) were below the normal range and FTI (5.8+/-0.6) was normal. All patients were male, and family studies revealed decreased TBG capacity in blood relatives consistent with X-chromosome linked inheritance. All examined relatives relatives of the propositi, whether hemizygous or heterozygous, were euthyroid. Over the same period of time 7 additional patients (excluding family members of propositi) were found to be euthyroid but had decreased TBG capacity. The occurrence of thyrotoxicosis in 5 out of 12 patients with inherited TBG deficiency suggests an association rather than a coincidental finding, although initial tests were performed because of suspected thyroid dysfunction. The incidence of thyrotoxicosis in patients with inherited TBG deficiency is also high on the basis of the reported prevalence of the latter genetic abnormality. This study stresses the importance of determining TT3 and FTI on patients who are clinically thyrotoxic but have normal TT4.


An FSH and prolactin-secreting pituitary tumor: pituitary dynamics and testicular histology.

A man with normal-sizes testes and a chromophobe adenoma of the pituitary was found to have elevated plasma levels of FSH and prolactin and severe deficiencies of other trophic hormones. Plasma FSH values doubled after administration of LH-releasing hormone but were not suppressed by exogenous testosterone. Prolactin concentrations were increased by TRH and were suppressed by L-dopa. Testicular histology revealed sparse and apparently inactive Leydig cells. Seminiferous tubules and Sertoli cells were well preserved, but there were few late spermatids.


Thyroid function in molar pregnancy.

Various aspects of thyroid hormone economy were examined in 15 patients with molar pregnancy and in 5 patients with choriocarcinoma. None of the patients with molar pregnancy was clinically thyrotoxic, though serum thyroxine (T4) was increased in 13 and free T4 concentrations were above normal in four of ten in whom measurements were made. In 2 patients with elevated serum T4 levels and in one patient with normal serum T4 levels, the daily rate of T4 disposal was increase, and most of 131I derived from 131I-T4 was excreted into urine in two patients in whom estimation of urinary excretion was made. Serum total triiodothyronine (T3) concentrations closely paralleled those of serum T4, but T3/T4 ratios were lower than those previously found in patients with toxic diffuse goiter of Graves' disease. Unresponsiveness to exogenous thyrotropin-releasing hormone (TRH) was observed in 5 patients with molar pregnancy whose serum total T4 and T3 levels were increased. Human chorionic gonadotropin (hCG) concentrations in serum ranged from 0.5 to 2830 IU/ml and were less than 200 IU/ml in 36% of the patients, possibly because molar pregnancy was diagnosed by ultrasonography, rather than high titers of urinary or serum hCG. Serum thyroid stimulating activity measured by the McKenzie bioassay closely paralleled values for hCG (r:+0.94). All thyroid abnormalities disappeared quickly after removal of the mole. In choriocarcinoma, no abnormality of thyroid function was found. These results suggest that: 1) some patients with molar pregnancy display a marked elevation of serum T4, T3, and free T4 concentrations, probably as a result of the action of a thyroid stimulator which is closely related to serum hCG or may be hCG itself; 2) T4 production rates are increased in these patients, and the pituitary responds to TRH in a manner similar to that found in thyrotoxicosis. Despite this, frank clinical thyrotoxicosis is usually absent, possibly because of relatively low serum T3/T4 ratios, possibly because of the limited duration of thyroid hyperfunction, and possibly because of other factors that for the present remain unknown.


The comparative effect of T4 and T3 on the TSH response to TRH in young adult men.

The effect of graded increments of chronically administered oral T4 or T3 on the TSH response to TRH was studied in normal young adult men. The TSH response was assessed in the baseline state and after each increment of each hormone (two weeks at each dose level) using both 30 mug and 500 mug doses of TRH. Each thyroid hormone caused a dose-related decrease in the TSH response to TRH; thus the TSH response could be used as a bioassay for the biologic activity of the thyroid hormones in man. The dose of thyroid hormone that caused a 50% suppression of the TSH response, or the SD50, was not different with either 30 mug or 500 mug of TRH indicating that thyroid hormone suppression of the TSH response is not more easily detected with a small dose of TRH. The mean SD50 for T4 was 115 mug/day, for T3 stopped 2 h before testing the mean SD50 was 29 mug/day, and for T3 stopped 24 h before testing it was 45 mug/day. Using the average SD50 for the two T3 regimens (37 mug/day), the calculated relative potency indicates that oral T3 is 3.3 times as potent as oral T4, a value in reasonable agreement with the value previously estimated with a calorigenic end-point. The mean dose of T4 needed to decrease the TSH response to TRH to below the normal range (max delta TSH of 2 muU/ml) was 150 mug/day; this value is probably more appropriate than the SD50 in the treatment of patients with primary hypothyroidism or goiter and was about the same (160 mug/day) using a peak TSH after TRH of 3 muU/ml as an end-point. Estimation of the SD50 in each subject showed a 2- to 3-fold range with all regimens of thyroid hormones; similarly there was a 2-fold in the dose of T4 needed to suppress the TSH response to TRH to below the normal range. Further, the difference in the mean SD50 for the two T3 regimens indicates that a single daily dose of oral T3 does not exert a constant biologic effect throughout the day. Thus, because of individual variation and, in the case of T3, because of changing activity during the day a given dose of thyroid hormone may have a widely varying biologic effect. There was also a 3-fold range in the relative potency of T3 to T4 in the four subjects treated with both hormones. This suggests that the therapeutic administration of a fixed ratio fo T3 to T4 may have a variable effect from patient to patient. Finally, the serum T4 rose while the serum T3 did not at a dose of T4 that abolished the TSH response to TRH, indicating that circulating T4 is a determinant of TSH secretion in normal man.


Serum levels of thyrotropin, thyroid hormones and their response to thyrotropin releasing hormone in infective febrile illnesses.

In 25 patients suffering from fever of infection, serum levels of thyrotropin (TSH), thyroxine (T4), triiodothyronine (T3), and thyroxine binding globulin (TBG) were estimated on two consecutive days during the febrile period and again 3 to 10 days after the fever had subsided. The serum TSH and T3 responses to 100 mug iv TRH were also studied during fever. Hormones were estimated by specific radioimmunoassays and TBG by radioligand binding assay. As compared with age and sex matched normal controls, patients with fever of infection had significantly lowered levels of total serum T3 and TBG. The serum TSH and total T4 concentrations were not significantly altered. During fever both % FT4 and absolute FT4 were significantly elevated, whereas only % FT3 was significantly increased and due to lowered serum total T3 levels the absolute FT3 were not significantly altered as compared to that in normal subjects. After the fever had subsided, the serum T3 levels returned to normal and the serum TBG levels increased. There was no correlation between basal serum levels of T3 and TSH during fever. Although in response to iv TRH the mean rise in serum TSH during fever was comparable to that in normal subjects, the overall TSH response showed an inverse correlation with serum TT3 levels. Following iv TRH there was a significant increase in serum T3 levels and the T3 response in fever was comparable to that in normal subjects. These data suggest that hormone secretion by the thyroid and its responsiveness to endogenous TSH are maintained during fever. The lowered T3 levels are not suggestive of a hypothyroid state, but perhaps could be due to decreased peripheral conversion of T4 to T3 and to decreased binding of T3 to serum proteins. The exact mechanism or significance of these alterations in thyroid function during febrile illness remains to be elucidated.


Apomorpine inhibits the prolactin but not the TSH response to thyrotropin releasing hormone.

Pretreatment of normal subjects with apomorphine, a dopamine receptor agonist, resulted in significant impairment of the  subsequent prolactin (PRL) response to thyrotropin releasing hormone (TRH). The mean maximal increment of PRL was 27.9+/-2.4 ng/ml after TRH alone, and 11.9+/-3.0 ng/ml (P less than 0.001) after apomorphine plus TRH. In contrast, the.thyrotropin (TSH) response to TRH was unaffected by apomorphine (10.5+/-2.9 vs. 9.5+/-1.8 muU/ml, P greater than 0.5). These results demonstrate that dopaminergic effects are capable of inhibiting PRL responses to TRH, probably via a direct effect on the lactotrope cell. They also suggest that dopaminergic influences are not important in the regulation of TSH secretion.


Studies on human chorionic gonadotropin secretion: effects of EGTA, lanthanum, and the ionophore A23187.

Human malignant trophoblast cells that secrete human chorionic gonadotropin (hCG) in culture were employed to assess the calcium requirement for hormone production. Cellular and secreted hCG was measured by radioimmunoassay. Cells cultured for 7 h in Ca2+- and Mg2-free medium or in Ca2+-free medium, secreted less hCG than cells cultured in medium containing Ca2+. Addition of ethylene glycol bis (beta-aminoethyl ether)N,N'-tetraacetic acid (EGTA), ethylenediamine tetraacetic acid (EDTA), or La2+ inhibited hCG release from the cells, but did not affect the amount of hCG in the cells. Inhibition of hCG release was dependent on time of incubation and on concentration of agent added. Inhibition of hCG secretion by EGTA was reversed upon removal of the EGTA from the culture fluid or by addition of equimolar Ca2+ to the fluid. These results demonstrated that divalent cations, probably Ca2+, are required for release and further synthesis of hCG. Addition of the divalent cation ionophore A23187 (0.1 to 10 muM) failed to increase hCG secretion by the malignant trophoblast, in contrast to the stimulatory effect of this agent in other secretory systems. Incubation of the cells for 15 to 60 min with 10 muM A23187 reduced hCG secretion, and this inhibition was reversed upon removal of the ionophore from the culture fluid. The studies with the ionophore supported other evidence indicating basic differences between hormone secretory mechanisms in the trophoblast compared to other endocrine tissues.


Age-related change in pituitary threshold for TSH release during thyroxine replacement therapy for cretinism.

Nine patients with athyrotic or ectopic cretinism ages 6 mo.-17 yrs-were studied to examine the relation between age and the quantities of oral L-thyroxine (T4) adequate to restore TSH hypersecretion to normal levels by feedback control. All but one patient had very low levels of endogenous T4 (below 1.0 mug/100 ml) before treatment. However, based on clinical signs and serum T4-1 values, all were judged to be euthyroid or sub-clinically hyperthyroid by L-T4 replacement during the observation period. L-T4 dosage was increased or decreased by 25 or 50 mug/day at 2-12 mo. intervals, and the TSH response to exogenous thyrotropin-releasing hormone (TRH) was tested sequentially at each dose of L-T4 until a normal or slightly suppressed TSH response was obtained The L-T4 dosage which was associated with normal TSH responsiveness to TRH (the adequate L-T4 dose) was high in infancy (10 mug/kg/day), decreasing with age to a level of 3-4 mug/kg/day in pubertal children (correlation coefficient r=-0.820, P less than 0.01). The adequate L-T4 dose observed between 4 and 12 yrs of age was lower than the usually recommended dose (4-6 vs. 7-8 mug/kg/day). On these L-T4 doses, serum T4I concentrations were significantly higher in patients under 5 yrs of age than in older patients (8.3+/-1.5 vs. 6.5+/-0.7 mug/100 ml, P less than 0.02). Serum triiodothyronine (T3) values were also elevated in young children, decreasing with age to the normal range. TSH responses to TRH were completely suppressed at the serum T4-I levels of 9 mug/100 ml and T3 of 250 ng/100 ml or less in 7 patients over 4 yrs of age, but not in 2 younger patients. Although the number of patients examined was limited, these results suggest that the pituitary threshold for feedback regulation of TSH secretion by T4 decreases with age in children with cretinism and the usually recommended replacement doses of L-T4 between 4 and 12 yrs of age are probably overestimated.


Immunoreactivity and serum destruction of N3immethyl-trh.

Pyroglu-N3immethyl-histidyl-prolineamide (MeTRH) is a more potent stimulator of thyrotropin and prolactin secretion than thyrotropin-releasing hormone (TRH). In this study, the immunoreactivity of MeTRH and its susceptibility to destruction by serum were investigated. When tested in radioimmunoassays employing four different anti-TRH sera. MeTRH had 25 to 60% of the immunoreactivity of TRH. Serum destroyed theimmunoreactivity of MeTRH at one-half of the rate at which TRH was destroyed. Slow degradation alone would not appear to explain the potency of MeTRH.


Immunological and immunoassay studies of the binding protein for vitamin D and its metabolites in human serum.

This study reports the development of a specific and sensitive radioimmunoassay and a simple and accurate radial immunodiffusion (RID) assay for the human serum-binding protein for vitamin D and its metabolites (DBP). These immunoassays employed a monospecific antiserum that was prepared in rabbits against human DBP. The radioimmunoassay effectively measured DBP in amounts of 1-10 ng, whereas the RID assay measured DBP accurately in amounts of 0.2-0.8 mug. The results obtained with the two immunoassays on the same samples of serum agreed well with each other. Using the RID assay, the mean (+/- SD) serum DBP concentration observed in 35 normal persons was 422 +/- 27 micrograms/ml. Generally similar levels were observed in 66 hyperlipidemic subjects. In molar terms, the mean DBP concentration (approximately 8 microgramsM) was of the order of 50 times the usual serum level of 25-hydroxyvitamin D (25-OH-D) plus vitamin D. Thus, most of plasma DBP circulates as apo-DBP, not containing a bound molecule of 25-OH-D or of vitamin D. DBP and 25-OH-D concentrations were measured in a limited number of patients with hypercalcemia, mild hypocalcemia, and markedly elevated serum 25-OH-D levels due to oral vitamin D supplementation. It was found that major changes can occur in the serum levels of 25-OH-D and of calcium with very little or no associated changes occurring in the serum concentration of DBP, The results suggest that neither serum 25-OH-D nor serum calcium plays an important role in the regulation of the metabolism of DBP. Data were obtained that confirmed and extended an earlier report on the identity of the group-specific component (Gc) protein in plasma with the plasma vitamin D-binding protein. On immunodiffusion against whole serum, the line formed with the anti-DBP antiserum showed a complete reaction-of-identity with the line formed with commercial antiserum against Gc protein. Furthermore, serum that had been depleted of DBP by treatment with Sepharose containing covalently coupled antibodies against DBP was found to be depleted also of immunoreactivity against anti-GC protein antiserum. In addition, the properties of the purified DBP preparation agreed closely with those previously reported by others for Gc protein. Finally, a comparative immunology study showed that sera from several different mammalian orders showed some immunoreactivity against the antihuman DBP antiserum. Thus, proteins immunologically similar to human DBP are present in sera from a number of mammalian species and orders.


Distribution of branched-chain alpha-keto acid dehydrogenases in primate tissues.

The specific activity of the branched-chain alpha-keto acid (BCKA) dehydrogenases was measured in normal tissues of the rat, monkey, and man, and in cirrhotic human liver. In the rat, specific activity of the dehydrogenases in liver, kidney, and muscle averaged 33, 26, and 0.4 U/g wet tissue, respectively; proportion of the body's content of the enzyme located in these three organs was 70, 12, and 10%. In the monkey, specific activities in liver and kidney were only one-half to one-third as great as in the rat, whereas activity in muscle was the same; the monkey's body content of dehydrogenase was distributed 50% in liver, 13% in kidney, and 20% in muscle. In man, specific activities in liver and kidney were only 1/15th to 1/25th as great as in the rat, but activity in skeletal muscle was the same. Distribution of the dehydrogenases in man was 30% in liver, 2% in kidneys, and 60% in muscle. In six patients with alcoholic cirrhosis, specific activity of the dehydrogenase in liver was reduced to 20-50% of normal (average, 32%). This reduction may alter the efficiency of BCKA as substitutes for branched-chain amino acids when BCKA are administered orally, but will have little influence on efficiency when they are given intravenously.


Relationship of aggregated intramembranous particles to water permeability in vasopressin-treated toad urinary bladder.

It has been previously demonstrated with freeze-fracture electron microscopy that vasopressin induces specific structural alterations of the luminal membrane of granular cells from toad urinary bladder in a dose-dependent fashion. These alterations consist of aggregated intramembranous particles and are observed both in the presence and absence of an osmotic gradient. We examined the effect of methohexital, a selective inhibitor of vasopressin-stimulated water flow, and the effect of phloretin, a selective inhibitor of urea permeability, on the structure of the granular cell luminal membrane. Methohexital treatment of the vasopressin-stimulated toad bladder reduced both the osmotic water flow and vasopressin-induced alterations of membrane structure to the same extent. Phloretin reduced urea permeability but not water flow or particle aggregation. Since neither agent affects vasopressin-stimulated sodium movement, these findings indicate that the phenomenon of particle aggregation is specifically related to vasopressin-induced water permeability and not to changes in urea or sodium permeability.


An ultrasonic multitransducer scanner for real-time heart imaging.

An ultrasonic multitransducer scanner has been developed. It works in combination with a conventional echocardiograph and is capable of instant dynamic cross-sectional imaging of the heart. A 10 cm wide area is scanned by 50 parallel sound beams, spaced 2 mm apart, at a frame rate of 36 per second. The equipment is described and preliminary clinical experience is reported. The scanner has been used for routine heart examinations and for obstetrical and abdominal scanning as well.


Gray scale measurement of biparietal diameter.

Fetal biparietal diameter measurements were obtained simultaneously by gray scale and bistable ultrasonography. The bistable presentation was measured from leading edge to leading edge, while the gray scale presentation was measured from leading point to leading point and middle to middle of the band of echoes defining the fetal head. Statistical analysis of 50 patients demonstrated good reproducibility of the biparietal diameter measurement by each method and intraobserver variation fell well within the acceptable range. In all but one patient differences in measurements between observers and measurement methods were trivial and of no clinical significance. In the patients in whom there was a statistically significant difference, the variability was from 1 to 2 mm with the exception of one patient in whom the variability was about 4 mm.


Oscillation of left ventricular structures in aortic regurgitation.

High-frequency diastolic oscillation of the anterior mitral leaflet is a common echocardiographic feature in aortic regurgitation. We have studied a group of 56 patients with aortic regurgitation in whom oscillation of the anterior mitral leaflet was present. Thirty-one of these had similar diastolic oscillation of the posterior leaflet; occasional patients demonstrated oscillation of the interventricular septum or chordae tendineae. The presence of these diastolic oscillations did not correlate with the presence of the Austin-Flint murmur. We conclude that diastolic oscillation of left ventricular structures other than the anterior mitral leaflet can be recorded on echocardiograms of patients with aortic regurgiationmthe mechanism responsible for these oscillations is not known; several possibilities are discussed.


Gray scale sonographic anatomy of the pancreas.

With the development of gray scale, the appearance of pancreatic anatomy and its relationship to surrounding structures can be demonstrated more effectively. Gray scale has made it possible to identify such vessels as the portal vein, superior mesenteric vein, superior mesenteric artery, splenic vein, etc. This makes it possible to identify the pancreas more precisely. The normal pancreas appears to have a fairly specific echo pattern. The pancreas scanning techniques used in this study include the oblique, transverse, and longitudinal sections. Oblique views are especially useful.


Ultrasound in the evaluation of adrenal metastases.

Metastatic involvement of the adrenal glands is a frequent complication of bronchogenic carcinoma, occuring in up to 33% of these patients. Examination of ten patients with adrenal metastases revealed large adrenal masses, occuring bilaterally in five. Five of these patients were examined using the standard bistable technique. These patients demonstrated largely echofree masses with only occasional scattered internal echoes. The remaining five patients were examined with the gray scale technique. With the added resolution afforded by the use of gray scale, the masses in these patients showed a more uniform distribution of echoes. Presence of a margin between the kidney and the mass, and displacement of the kidney inferiorly, posteriorly, and laterally is used to differentiated between upper pole renal masses and those of adrenal origin. Autopsy confirmation was obtained in six of the patients.


The antepartum findings of conjoined twins.

Various estimates abound as to the incidence of conjoined (Siamese) twins, but they are indeed uncommon. The probability of their occurance has been calculated to be once in every 12,000 to once in every 100,000 deliveries. Although uncommon, the antepartum diagnosis can be extremely important to the twins and the mother, if only to spare the trauma of a trial by labor. The findings, both radiographic and ultrasonic, are presented herein.


Extra-amniotic blood clot simulating placenta previa on ultrasound scan.

The ultrasonic appearance of extra-amniotic blood clot simulating placenta previa is presented. The single case presented is the most illustrative and best documented example of this phenomenon in a series of 10 patients. The ultrasonic characteristics of this phenomenon are discussed.


Echocardiographic appearance of aortic valve vegetations in bacterial endocarditis due to Actinobacillus actinomycetemcomitans.

This report describes the echocardiographic features seen in a case of bacterial endocarditis due to Actinobacillus actinomycetemcomitans. The echocardiogram demonstrated large, bulky clusters of echoes filling the aortic root in diastole which extended to the subvalvular and supravalvular areas. The echocardiographic features mimicked those described in cases of Candida endocarditis. The morphologic appearance of vegetations at surgery correlated with the echocardiographic findings.