Sensitivity and specificity of electrocardiographic diagnosis of ventricular motion disorders. Studies in patients recovered from myocardial infarction. Angiographic-electrocardiographic correlations were studied in 253 patients with coronary arterial disease. Left ventriculograms were examined for various motion disorders (local hypokinesia, akinesia, dyskinesia or aneurysm, and generalized hypokinesia). Electrocardiograms were reviewed for the presence of Q waves indicative of transmural myocardial infarction and for the presence of upward convex ST-segment elevation of at least 2 mm. Of 27 consecutively studied subjects with normal coronary arteries and normal ventriculograms, none had Q waves, giving a specificity of 100% for the Q wave. Pathologic Q waves (greater than or equal to 0.04 second) were found in 20% (19) of 96 patients with coronary arterial disease having normal ventricular motion, reducing the specificity for the Q wave to 80% in patients with documented coronary disease. Of 157 patients with coronary disease and ventricular dyssynergy, 118 had pathologic Q waves, yielding a sensitivity of 75%. Concomitant Q waves plus 2 mm ST-segment elevation were present in 45 of the 118 postinfarction patients with ventricular dyssynergy, giving a sensitivity of 38%. The specificity of this combination in coronary patients was 84%.


Ultrastructural morphometry of the blood-air barrier in pulmonary sarcoidosis. Stereologic techniques were utilized in an electron microscopic study of biopsy samples obtained from the lungs of seven patients with pulmonary sarcoidosis. Relative fractional volumes of alveolar septal components and the arithmetic mean thickness and harmonic mean thickness of alveolocapillary membranes (blood-air barrier) were compared with values for normal lungs. Based on morphometric analysis, increases in the arithmetic mean thickness and the harmonic mean thickness of the alveolocapillary membranes appeared too small to account for the reduction in gas transfer present; however, there was a quantitative relative increase in interstitial tissue in alveolar septa, which does not take part in gas exchange, at the expense of the capillary bed, which is critical to this function.


Electrocardiographic radiotelemetry in the early recuperative period of acute myocardial infarction. Its value in detecting arrhythmias in the setting of a progressive coronary care unit. One hundred seventeen consecutive patients with acute myocardial infarction transferred from the coronary care unit to a progressive coronary care unit were monitored by electrocardiographic radiotelemetry for a median of five days for arrhythmia detection. Eighty patients (68%) had arrhythmias; 37 patients (32%) had none. Almost all arrhythmias were detected by the sixth day of monitoring. Thirty-nine patients (33%) had premature ventricular contractions, two patients had primary ventricular tachycardia, one patient had primary ventricular fibrillation, and two patients had secondary ventricular fibrillation. An additional patient was successfully resuscitated after monitoring had been discontinued. Four (3%) of 117 patients had a potentially lethal arrhythmia treated successfully, three while being monitored. Twenty-five of the patients (21%) had 33 episodes of treatment intervention because of telemetrically detected events. Five patients (4%) died, all of unpreventable causes. No patient died because of an arrythmia. We conclude that telemetry is feasible and practical in detecting arrhythmias. It is clear that potentially fatal ventricular arrhythmias do occur following transfer from the coronary care unit and that monitoring is, therefore, desirable.


Effect of oxygen in graded concentrations upon tracheal mucous velocity. A study in anesthetized dogs. Graded concentrations of oxygen were used to establish dose-duration relations for the effect of oxygen on tracheal mucous velocity and tracheobronchial histologic findings in the anesthetized dog. Observations of tracheal mucous velocity were made during 30-hour periods of breathing air (100-percent humidified and warmed to 38 degrees C) and oxygen mixtures (also 100-percent humidified and warmed to 38 degrees C). In animals breathing oxygen mixtures, the baseline tracheal mucous velocity was taken as the value while breathing room air at the experiment's start. No statistically significant differences in tracheal mucous velocity occurred during air breathing. Tracheal mucous velocity fell 45 percent from the baseline value after breathing 100-percent oxygen for two hours (P less than 0.01), fell 42 percent after 75-percent oxygen for nine hours (P less than 0.01) and fell 51 percent after 50-percent oxygen for 30 hours (P less than 0.001). Histologic examination of the trachea and major bronchi after six hours of 100-percent oxygen and 12 hours of 75-percent oxygen revealed signs of acute tracheobronchitis. Minor histologic alterations in the tracheobronchial tree occurred both in animals breathing air and 50-percent oxygen for 30 hours; therefore, histologic evidence of oxygen toxicity could not be established at 30 hours. These findings indicate that in the anesthetized dog, oxygen depresses mucous transport as a function of inspired oxygen concentration (FIO2) and that even FIO2 as low as 0.05 might be deleterious.


Detection of unstable angina by 99m technetium pyrophosphate myocardial scintigraphy. 99mTechnetium stannous pyrophosphate has been shown to accumulate in acutely infarcted myocardium. To determine if the isotope is also taken up by severely ischemic, but not necrotic myocardium, we performed myocardial scintigraphic studies in 17 patients with chest pains. Seven of the patients satisfied conventional clinical, electrographic, and laboratory criteria for the diagnosis of unstable angina and showed no electrocardiographic or enzymatic evidence of myocardial necrosis. Five of these seven patients with unstable angina demonstrated abnormal localized patterns, and one showed a borderline picture. Myocardial scintiscans were normal in all of a control group of ten patients with stable angina. Thus, scanning with 99mtechnetium stannous pyrophosphate is shown to be of value in the objective demonstration of myocardial abnormality in unstable angina.


Chest physiotherapy and the distribution of ventilation. The injured or postoperative thorax is commonly treated with physiotherapy employing "unilateral breathing" techniques. The ability of these methods to alter the distribution of ventilation and blood flow (oxygen uptake) was examined in four patients undergoing bronchospirometry. No change was observed in these measurements in the lung toward which efforts were being directed to either enhance or restrict the mechanics of breathing. To further examine the problem, two susceptible subjects were hypnotized following intubation for bronchospirometry, and emotional and physical states were suggested that limit motion in one hemithorax. This included the presence of pleuritic pain in one subject who had ample experience with this symptom. In no instance did we find a change in the distribution of ventilation or blood flow to the lung that was supposed to be limited; however, in 17 subjects, lying on one side produced a significant increase in ventilation and blood flow in the dependent lung. Lying on one side or the other consistently and predictably modified ventilation and blood flow, while attempts to produce similar changes by voluntary intention or commonly applied mechanical aids were ineffective.


Comparison of tracheobronchial suction catheters in humans. Visualization by fiberoptic bronchoscopy. Tracheobronchial suctioning was performed in 20 patients undergoing diagnostic or therapeutic bronchoscopic examination. The effect of a single suctioning procedure on the airway mucosa was observed and recorded by still and cine photography through a fiberoptic bronchoscope. A variety of catheters of various designs were evaluated. All produced a negligible amount of trauma, and none was superior in efficiently evacuating mucus from the airways. Mucosal trauma with tracheobronchial suctioning procedures is more likely due to repetition, vigor, and amount of suction applied, regardless of which type of catheter is used.


Stenotic semilunar valve in persistent truncus arteriosus. The clinical, hemodynamic, and pathologic findings in two newborn infants with persistent truncus arteriosus and stenosis of the truncal valve are described. In one case the anatomic features of the basic condition were classic, with a dysplastic semilunar valve which was mainly stenotic and also incompetent, while in the other the truncus arteriosus arose exclusively from the right ventricle and was almost exclusively stenotic. A ventricular septal defect was the only outlet for the left ventricle. In this case, mitral stenosis was also present and associated with a left-to-right shunt at the atrial level.


Corrected transposition of the great arteries simulating coronary heart disease in adults. Two middle-aged adults with corrected transposition of the great arteries presented with chest pain and dyspnea, initially attributed to coronary arterial disease with left ventricular dysfunction and mitral regurgitation. The clinical diagnosis of coronary arterial disease was supported in these patients by electrocardiographic changes of probable old anteroseptal myocardial infarction and by an abnormal cardiac silhouette on the chest x-ray film suggestive of ventricular aneurysm. Coronary arteriographic studies in both patients showed that the morphologic left coronary artery supplied the venous ventricle, and the right coronary artery supplied the systemic ventricle. Neither patient had coronary arterial luminal narrowing. We propose that anginal chest pain in these patients resulted from inadequate coronary reserve of the morphologic right ventricle functioning as a systemic ventricle.


Right ventricular apexangiography for precise placement of transvenous electrode catheters. A simple angiographic technique (right ventricular apexangiography) for positioning permanent transvenous pacemakers is described. With this technique, electrode failure occurred in only five (3%) of 186 patients followed for more than two years, as compared to an average failure rate of 23% in 11 reported series. Right ventricular apexangiography should be particularly useful in those patients in whom problems arise during insertion of a permanent transvenous electrode catheter.


Alveolar proteinosis. Lobar lavage by fiberoptic bronchoscopic technique. Lavage limited to an isolated lobe was performed on multiple occasions using a cuffed fiberoptic bronchoscope in a patient with alveolar proteinosis. Sequential ventilation-perfusion scintiphotoscans were used to preselect and follow the functional behavior of the lavaged lobe. Lavage led to functional improvement. The technique of fiber-optic bronchoscopic lobar lavage is simple and may find application in patients in whom lavage of an entire lung may be hazardous.


Melioidosis complicated by pericarditis. A case of acute and recrudescent melioidosis complicated by pericarditis and pericardial effusion is described. The potential for the appearance of future cases in the United States and the necessity for physicians to remain aware of this potential diagnosis are discussed.


Tumor emboli presenting as pulmonary hypertension. A diagnostic dilemma. A case of recurrent tumor emboli secondary to choriocarcinoma is described. The patient presented with obvious pulmonary hypertension and was diagnosed and treated as a case of multiple pulmonary embolism. Information which suggested the possibility of tumor emboli was indeed present but recognized only retrospectively.


Median sternotomy wound infection and anterior mediastinitis caused by bacteroides fragilis. The first reported case of median sternotomy infection and mediastinitis caused by Bacteroides fragilis is described. Several positive blood cultures led to diagnostic anaerobic cultures of the wound and administration of clindamycin therapy. This anaerobic infection should be suspected whenever Gram smears of exudates show gram-negative bacilli and aerobic cultures are sterile.


Diagnosis of inferior wall myocardial infarction during right ventricular apical pacing. The diagnosis of inferior wall myocardial infarction is often masked during ventricular pacing. We observed paced ventricular beats with a qR pattern in leads 2,3, and aVF in a patient with acute inferior wall myocardial infarction and a temporary pacemaker at the apex of the right ventricle. Such a pattern might be specific for the diagnosis of inferior wall myocardial infarction, because it is never seen during uncomplicated pacing from anywhere within the right ventricular cavity.


Chronic calcified aortic dissection.

Chronic calcified aortic dissection. Two cases of chronic aortic dissection are presented. In both, plain chest roentgenograms revealed calcification in the outermost aspect of the dilated aorta. It is clear that this represented calcification of the dissection rather than of the true intima. These cases indicate that apparent aortic intimal calcification with dilatation of the aorta is not completely diagnostic of an arteriosclerotic aneurysm and that a chronic calcified aortic dissection must also be considered in the differential diagnosis.


Inferior myocardial infarction in a patient with mirror-image dextrocardia and situs inversus totalis. A case of inferior wall myocardial infarction in a patient with mirror-image dextrocardia and situs inversus totalis is presented. The clinical and electrocardiographic findings are discussed.


Transatrial closure of ventricular septal defect in corrected transposition of the Great Arteries. The repair of ventricular septal defect (VSD) in corrected transposition of the great arteries has presented many technical problems and was associated with a high mortality. Transatrial closure of the VSD in this malformation obviates many of the hazards and offers good exposure. The operative procedure in such a case is described in detail, and its merits are discussed.


In situ hybridization of "nick-translated" 3H-ribosomal DNA to chromosomes from salamanders. A technique is described for preparation of 3H-labelled DNA by nick-translation employing deoxyribonuclease I and DNA polymerase I. The labelled DNA can be obtained in high yield with specific activities of 10(6) cpm/mug or more. Ribosomal DNA, isolated from ovaries of young Xenopus laevis, and whole DNA from Plethodon cinereus were labelled in this way. The rDNA was used for in situ hybridization to meiotic chromosomes from P. cinereus, P. vehiculum and P. dunni. Autoradiographs of in situ hybrids were exposed for 5 to 10 days, by which time nucleolus organizer regions on the chromosomes of all 3 species were clearly and specifically labelled. In all cases, labelling was confined to a short region near the middle of the short arm of both halves of a medium length bivalent. It is concluded that nick-translation is a useful and altogether efficient method of labelling nucleic acids for subsequent use in experiments involving in situ hybridizations.


In situ detection of a DNA-polymerase activity in the nuclei of mouse spermatozoa. A nuclear DNA-polymerase activity has been detected in situ in mouse spermatozoa with a cytochemical method. The acid-insoluble radioactive product obtained after incubation in a mixture containing all four deoxyribonucleoside 5'-triphosphates, Mg++,KCl and dithiothreitol (DTT) was completely removed by DNAse but was insensitive to RNAse and pronase action. The polymerization reaction did not take place in the presence of only one nucleotide and was dramatically reduced when actinomycin D was present; it did not seem to be inhibited markedly by N-ethylmaleimide (NEM) or by parahydroxymercuribenzoate (PHMB) but was sensitive to high concentrations of KCl.--The reaction depended strictly on the presence of nuclear DNA as template since treatment with pancreatic DNAae before the DNA-polymerase assay completely prevented the appearance of any radioactivity.


Further studies on polyploid amphibians. V. C-banding in diploid and tetraploid species of Odontophrynus. C-band patterns of some Odontophrynus species from Brazil are described. O. americanus 2n and 4n are cryptic species similar to O. cultripes 2n, but their C-banding patterns are different. In O. americanus 2n the C-band appears at the secondary constriction of chromosomes 4. In O. americanus 4n and O. cultripes the C-band appears at the secondary constriction of chromosomes 11. O. cultripes chromosomes 2,3 and 6, have also an intercalary band. Furthermore, fine centromeric bands in several weeks old preparations of these species were observed. The possibility of a structural or chemical heterogeneity of those heterochromatic regions is discussed.


Karyotypic variation in the Australian gekko Phyllodactylus marmoratus (Gray) (Gekkonidae: Reptilia). The gekko Phyllodactylus marmoratus has at least three distinct chromosome races; 2n=39, 2n=26 ZZ/ZW and 2n=34. Specimens from these races are morphologically distinguishable, have a degree of habitat specialization and occur in a defined distribution. The 2n=36 race found in Eastern Australia is the presumed primordial type. The 2n=34 race occurs in Western Australia and is regarded as a fusion derivative. The 2n=36 ZZ/ZW race, which is only found on the Murray River system in Eastern Australia has a heteromorphic sex chromosome system present in the female. Giemsa banding suggests that this heteromorphism is the result of a pericentric inversion.


Pairing and segregation of the sex chromosomes in X1X2-males of Dysdercus intermedius with a note on the kinetic organization of heteropteran chromosomes. The existence of an X1X2-mode of sex determination is confirmed by a study of all meiotic stages in the male cotton stainer (X1X2) and pertinent stages in the female (X1X1X2X2). In the male, the X-chromosomes are heterochromatic and pair end-to-end in early meiotic prophase. At diakinesis, they disjoin and align side-by-side in the center of the spindle, forming a pseudotetrad. Anaphase I is equational for the sex chromosomes. At late anaphase or telephase, X1 and X2 join end-to-end but form spindle fiber connections to only one of the poles of the metaphase II spindle, leading to one daughter cell without X chromosomes and one with both X1 and X2. An attempt is made to explain sex chromosome pairing and orientation on the basis of a telocentric organization of meiotic chromosomes. The apparent differences in the kinetic organization of mitotic and meiotic chromosomes in Heteroptera are discussed.


Interchromosomal asynchrony of DNA replication in polytene chromosomes of Drosophila pseudoobscura. Analysis of 3H-thymidine autoradiograms of late third instar larval salivary glands of Drosophila pseudoobscura revealed a unique example of asynchrony of replication in the autosome complement. The two autosomal arms, 2 and 3, show similar labeling pattern during the initial phases, DD to 3C, and thereafter, the chromosome 3 has fewer labeled sites than chromosome 2 until the most terminal pattern, 1D. Detailed sitewise analysis of 3H-thymidine labeling shows that while nearly 54% of the sites examined in chromosome 2 have a labeling frequency greater than 50%, only 13% of all sites in chromosome 3 have labeling frequency at that range. The number of labeled sites on chromosome 3 plotted against that on chromosome 2 shows a hyperbolic profile rather than a linear relationship. The silver grain ratio of the 2nd to 3rd increases from 1.5 to 3.1 through different stages of the cycle. These results suggest that both chromosomes start replication simultaneously but the third chromosome appears to complete the replication earlier than the second. These data open up the possibility of separate control mechanisms for the initiation and termination of DNA replication in polytene chromosomes.


Patterns of incorporation of tritiated thymidine by the dorsal polytene foot-pad nuclei of Sarcophaga bullata (Sarcophagidae:Diptera). The incorporation of tritiated thymidine into the DNA of the dorsal foot-pad nuclei of Sarcophaga bullata, during pharate adult development, was studied by scintillation counting and autoradiography. Incorporation was maximal on day 4 and showed a progressive temporal decline on days 5 to 8. Autoradiographs of chromosomal arms A1, A2 and D1, from late stages of the prolonged S period, showed discontinuous incorporation. On days 7 and 8 loci which incorporated the radio-labeled precursor were almost exclusively associated with DNA granules. DNA granules were associated with 8 specific loci in chromosomal arm A1, 10 in A2, and 8 in D1. The relationship between patterns of incorporation of tritiated thymidine and DNA granules is discussed.


Chromosome homology and evolution of emydid turtles. G-, C-, Q-banding and standard karyotypic analyses were used to study the chromosomal relationships of emydid turtles. Ten species of emydids were used (5 batagurines and 5 emydines) which samples all of the karyotypic variation known for the Emydidae. Data from a testudinid and a chelydrid are compared to the emydids. The karyotype of Mauremys and Sacalia is considered representative of the primitive karyotype for this group because of its widespread occurrence in the morphologically primitive Batagurinae and its similarity to that of some testudinids. The emydine karyotype is believed to have evolved from the primitive batagurine karyotype by the deletion of a heterochromatic macrochromosome. Siebenrockiella and Rhinoclemys are karyotypically derived batagurines.


Population cytogenetics of the genus Caledia (Orthoptera: Acridinae). I. Inter- and intraspecific karyotype diversity. The genus Caledia acontains two species. C. species nova 1 is restricted to the Oriomo Plateau of S. W. Papua and has a complement of twelve telocentric chromosomes. The second species C. captiva has a much wider distribution pattern--from S.W. Papua in the North, down the entire Eastern seaboard of Australia to Southern Victoria. It is also found in the Northern Territory. Although the chromosome number is the same as C. species nova 1, four major and distinct chromosomal races can be distinguished in C. captiva.--The basic "ancestral" race is found in Tropical North Queensland at the base of the Cape York Peninsula. All twelve chromosomes are telocentric and the karyotypic organization is similar to that found in C. species nova 1 and in other Acridines. A second, "general purpose" karyotypic race has a wide distribution between S. W. Papua, Arnhem Land and the East Australian coast as far South as Brisbane. It is considered a derivative form of the "ancestral" type and is fixed for small pericentric inversions on seven pairs of chromosomes. In the South-Eastern Queensland region there exists a further race which carries large pericentric inversions on all the autosomes and the X chromosome. The situation here is confounded since the basic chromosomes can be represented as either acro- or telocentrics. Various levels of polymorphism for the inversions exist between different chromosomes in different populations indicating considerable differentiation within this zone. This race is almost completely surrounded by the "general purpose" karyotype where the races are contiguous in certain parts of the range.--The South-Eastern corner of Australia is characterised by a chromosome race quite different from those found further North. Here a complex pericentric inversion system exists involving a series of seven small inversions and larger inversions on chromosomes 1, 2, 4 and 10. Chromosomes 2 and 4, in particular, are highly polymorphic.--The presence and persistence of these 4 chromosomal races can be accounted for in terms of the known climatic changes which have occured in this region in the recent past.


The meotic prophase in Bombyx mori females analyzed by three dimensional reconstructions of synaptonemal complexes. Serial sectioning followed by three dimensional reconstruction of lateral componenets of the synaptoemal complex have been used to follow chromosome pairing during theprophase of the achiasmatic meiotic division in the silkworm.  Bombyx mori.  During leptotene and early zygotene, the lateral components become attached to the nuclear envelope at a specific region, thus forming a chromosome bouquet.  The attachment of lateral componenets  to the nuclear envelope precedes the completion of the components between their attachment points.  Synapsis and synaptonemal complex formation start during the period of lateral component organization in the individual nucleus.  Telomeric movements on the nuclear envelope occur at two stages of the prophase:  the chromosome pairing appears to be initiated by an association of unpaired ends of homologus chromosomes, the nature of this primary attraction and recognition being unknown.  Secondly, the paired chromosomes become dispersed in the nucleus by shifting of attachment sites of completed synaptonemal complexes at the end of zygotene.  This movement is possible related to a membranes flow occuring during this stage.  Membrane material is synthesized at the region of synaptonemal complex attachment.  Later, the excess membrane material is shifted to the opposite pole where it protudes into the lumen of the nuclei thus forming vacuoles.  Two previously undescribed features of chromosomes paring were revealed.  In late zygotene, chromosome pairing and synaptonmal complex formation were frequently observed to be delayed or even prevented over s short distance by interlocking two bivalents, both being attached to the nuclear envelope.  Such interlocking of bivalents was not found in pachytene...


Anal fistula: a review of 82 cases.

Anal fistula: a review of 82 cases. Eighty-two cases of anal fistula have been retrospectively reviewed, and their clinical presentations, pathology, and management presented. Most of the fistulas were primary and, except for the four cases due to tuberculosis, there does not seem to have been any underlying systemic or gastrointestinal causative factor.  A preoperative fistulogram is essential in the management of this condition.


[Clinical significance of the 14C-glycocholate breath test in the diagnosis of gastro-enterological diseases (author's transl)]. The 14C-glycocholate breath test was performed in 15 normal subjects and 134 patients clinically suspected of bacterial overgrowth in the proximal small intestine, with functional impairment of the ileum and chologenic diarrohea as well as other forms of diarrhoea. In addition, faecal weight, faecal fat excretion and faecal bile-acid excretion were measured. Early and highest 14CO2 expiration peaks were found as an expression of increased deconjugation of bile acids in patients with fistulae between proximal small intestine and colon, and in 13 of 24 patients with Billroth II gastric resection or duodenopancreatectomy. Bile-acid deconjugation was not increased in sprue, chronic pancreatitis with steatorrhoea, ulcerative colitis, irritable colon, Whipple's disease, Salmonella enteritis, non-specific enteritis, or laxative abuse. In six of twelve patients with Crohn's disease of the ileum there was an increase in deconjugation of bile acids.


[Cogan's syndrome with angitis of cranial nerves, aortitis, endocarditis, and glomerulonephritis (author's transl)]. A 34-year-old woman died of left-heart failure due to combinated aortic-valve disease three years after manifestation of Cogan's syndrome characterized by sudden inner-ear deafness, loss of equilibrium, interstitial keratitis, and progressive loss of vision during pregnancy. At necropsy there was evidence of recurrent endocarditis of the aortic valves with stenosis and regurgitation, severe angitis of the thoracic aorta with marked secondary sclerosing changes as the cause of the heart failure. Angitis within the optical fasciculus and stato-acoustic nerve was the cause of the vestibular and optical defects. Primary changes in the visual and auditory cortices, the retina and inner ear were excluded as causes. There was also acute membrano-proliferative glomerulonephritis, which may have been coincidental.


[Cytological investigations of the endometrium using the jet wash technique (author's transl)]. In 182 cases the jet wash technique proved very suitable in the detection of malignant changes in the endometrium. It should in no way replace the curettage as a diagnostic tool. The jet wash offers the possibility of a regular endometrial investigation to larger numbers of patients. Those to be considered in particular are 1. symptom-free women with an increased risk of carcinoma of the body of the uterus, i.e. women over 40 with obesity, hypertension and diabetes mellitus, 2. patients with recurrent bleeding who have already had a curettage with negative histological results, 3. primary irradiated endometrial carcinoma patients in order to detect a recurrence early, 4. patients who pose a high anaesthetic risk. The exact detection reliability of the endometrial jet wash technique remains uncertain until results of larger investigation series are published. Good experience up to now with the jet wash technique should stimulate its wider use.


[Charcoal haemoperfusion in the treatment of sleeping-drug intoxication (author's transl)]. Extracorporeal haemoperfusion is indicated in the treatment of severe intoxication with sleeping-drugs (including barbiturates) if other methods have failed. Four patients in whom haemoperfusion was used (attempted suicide with hypnotics, grade III-IV and IV severity) were successfully resuscitated without permanent sequelae.


[Morphology and pathophysiology of Billroth I gastro-duodenostomy (author's transl)]. Endoscopic and radiological studies were undertaken in 97 patients who had had a partial gastric resection with gastro-duodenostomy (Billroth I) one to seven years previously. Acid secretion was also analyzed after injection of pentagastrin. The most frequent mucosal changes seen at endoscopy were erosion in the residual stomach, with inflammatory granulation in the region of the anastomosis. Recurrent ulcers were seen in 3%. Acid secretion was reduced by 87% from its pre-operative level. The amount of residual secretion depended on the length of the greater curvature in the residual stomach. Radiological studies revealed functional changes indicating that there is a motility pattern in the residual stomach and a pseudopyloric function at the anastomosis which makes co-ordinated emptying possible.


[Lesch-Nyhan syndrome without self-mutilation: biochemical and morphological studies on blood cells (author's transl)]. Absent enzyme activity of hypoxanthine-guanine-phosphoribosyl transferase in erythrocytes was demonstrated in the blood from a 21-year-old man with spastic tetraplegia, choreoathetosis, oligophrenia, and hyperruricaemia which developed after birth. But there was no tendency towards self-mutilation, considered to be characteristic of the full-blown picture of the disease. This case thus runs counter to the view that the enzyme defect is the cause of the inevitable, stereotyped change in behaviour. Unusual morphological changes of all blood-cell systems confirm the dependence of nucleic acid synthesis on the "salvage pathway" of the purine nucleotides, which is abnormal in this inherited disease.


[Q-Fever and its complications (author's transl)]. Q-Fever recurs as localized epidemics in the German Federal Republic. The disease has not previously been observed in North Hesse. During a familial infection with Coxiella burneti a 15-year-old boy together with his parents and an aunt became ill with atypical pneumonia. After the pulmonary symptoms had subsided a deep left leg and pelvic venous thrombosis occurred in the boy in the fifth week after the onset of the disease. The thrombosis which is very rare in childhood could be successfully removed at operation.


[cis-Diamino-dichloro-platinum (II) in the treatment of otherwise treatment-resistant malignant testicular teratoma (author's transl)]. cis-Diamino-dichloro-platinum (II) (DDP, NSC-119875) is an inorganic compound with cytostatic properties which have only recently been appreciated. DDP has an action which may tentatively be described as alkylating through some, as yet unknown, metabolic product. In a phase I study the effectiveness against testicular neoplasma had been established. A prospective trial was, therefore, conducted to test the drug's efficiency in patients with disseminated non-seminomatous testicular cancer refractory to all previously used combinations of chemotherapy. In a group of 22 patients the results were: 1 CR, 14 PR, 2 NC, and 5 PD. Because of these results, DDP was made part of an induction regime for disseminated and non-seminomatous testicular tumors.


[Toxic polyneuropathy after sniffing contact glue thinner (author's transl)]. Four men aged 16 to 19 years who had sniffed contact glue ("Pattex") thinner almost daily for 3 to 7 years developed a pronounced polyneuropathy. They had to be admitted nearly at the same time. A uniform neurological syndrome similar to Landry's paralysis with progressive ascending symmetrical pareses had developed. Motor deficiencies and atrophies affected the lower extremities more frequently and more severely. Only minimal sensory disturbances were found. The disease shows remarkable similarity to the "glue-sniffing neuropathy" described in the US and Japan which is attributed to n-hexane.


[Amyloidosis in familial mediterranean fever: clinical and renal-biopsy features (author's transl)]. In two Turkish female patients, aged 14 and 29 years, with familial mediterranean fever amyloidosis of the perireticular type was found. The disease was characterized by feverish bouts, abdominal colics, and joint involvements. The younger patient had the diagnosis confirmed at an early stage by renal biopsy, and under heparin and azathioprine the clinical signs, especially the nephrotic syndrome, regressed over a period of seven months. The second patient died of treatment-resistant shock in acute renal failure, due to rapidly progressing renal amyloidosis.


[Renal tubular failure after treatment with high dose of vitamin A (author's transl)]. A 22-year-old man was given 70-75 million I.U. vitamin A by mouth for 38 days as a treatment for psoriasis. But it had to be stopped because of the appearance of typical signs of vitamin A poisoning. Acute renal failure set in nine days later, after hospital admission for cerebral signs. In addition to conventional conservative treatment, seven haemodialyses were undertaken, with complete restitution to normal within 12 days. The skin disease and the cerebral signs predominated, in addition to later anaemia, hypercalcaemia, bone pain, and acute renal failure, the latter confirmed by renal biopsy. There were no histological changes in the liver.


[Treatment of acute cerebral ischaemia with low-molecular dextran: results of a retrospective study (author's transl)]. Between 1968 and 1970, 226 patients with acute cerebral ischaemia were treated with vasodilator drugs. In the subsequent three years (1971 to 1973) 202 patients were instead treated with dextran (Rheomacrodex) infusions. tin the latter period the death-rate was 35%, 19% less than during the preceding period of vasodilator treatment. In the dextran group among patients over 65 years the death-rate was 42%, 21% less than in the vasodilator group of the same age. Dextran treatment caused complete disappearance of neurological signs in 25%, compared with 7.5% in the vasodilator group. Dividing patients into those with normal blood pressure, hypertension and diabetes gave corresponding differences among the two groups. The death-rate during the period of dextran treatment was significantly less in all the sub-groups than in the vasodilator years. The results of this retrospective study provide important pointers to the effectiveness of dextran infusion in the treatment of acute cerebral ischaemia.


[A controlled study of long-term treatment of haemophilia B on an out-patient basis (author's transl)]. Eight patients with severe or moderately severe haemophilia B were treated for six months, according to three schemata for two months each: 18, twice 18, or twice 9 U of factor IX per kg body-weight weekly. The sequence of the six possible treatment schemes was determined strictly at random. One patient had to be excluded because he developed hepatitis, another because allergic signs developed. In the pre-trial period the number of bleedings per two months had been about 40, as recorded by the patients. The bleedings were reduced to nine after 18 U of factor IX per kg body-weight weekly, falling to two after twice 9 and twice 18 U/kg weekly. The patients themselves considered twice 9 U/kg as the ideal dosage. It is recommended that this dosage scheme should be used initially if one decides to employ permanent substitution in haemophilia B. Once freedom from bleedings and strengthening of the motor system have been achieved one can then try 18  U per kg body-weight once a week.


[Lactic acidosis after administration of buformine (author's transl)]. An 85-year-old female diabetic was admitted in coma, having been on antidiabetic treatment with the biguanide derivative "Silubin retard", 600 mg/d, for one-and-a-half months. The anion deficiency was 57 mmol/l, pH 6.9, suggesting the diagnosis of lactic acidosis in the absence of other causes of metabolic acidosis. Blood lactic acid levels of more than 16.65 mmol/l (150 mg/100 ml) confirmed the diagnosis. Administration of 875 mmol sodium bicarbonate over 12 hours corrected the deficiency. On admission to hospital there had been slight pre-renal failure. Myocardial infarction developed as a result of tissue hypoxia but did not prove clinically important. On the second day there were signs of a compensated disseminated intravascular coagulopathy with upper gastro intestinal haemorrhage. The woman died suddenly 18 days later of pulmonary embolism.


[Complications of acupuncture (author's transl)]. During acupuncture in the chest region a right-sided pneumothorax developed in a 53-year-old woman which necessitated emergency admission to hospital. A reinflation of the lung could be achieved with two Bülau drains. The increasing frequency of acupuncture must remind one of the feasibility of lesions of internal organs including those of the abdomen. Complications due to insufficient disinfection of acupuncture needles  may also arise.


[Weight reduction and glucose intolerance in obese subjects (author's transl)]. In a study extending over five years 35 of 70 excessively overweight patients with subclinical diabetes mellitus who had not significantly changed their weight, demonstrated a further deterioration in glucose tolerance, and manifest diabetes occurred in ten. In the remaining 35 patients who had achieved 20% weight reduction from the initial level glucose tolerance had become normal. The two groups were similar as to age, initial weight, and original degree of glucose intolerance. There was a significant correlation between loss of weight and increased glucose tolerance. No such correlation occurred with subgroups of patients whose initial overweight was more than +100 relative percentage and who had been over 50 years old at the start of the study.


[The diagnostic value of CK-isoenzymes in suspected acute myocardial infarction (author's transl)]. CK-isoenzymes were measured in 31 patients hospitalised for suspected myocardial infarctions who had an increase in serum creatine kinase (CK) above 50 U/l. Of 26 patients with definite evidence of myocardial infarction, MB-isoenzyme--specific for myocardial necrosis--was demonstrated in 24. MB-isoenzyme was no longer detectable in two patients hospitalised 48 hours after the onset of symptoms. In the remaining five patients only MM-isoenzyme was found, the elevated CK activity in three patients having been due to an intramuscular injection, and in two others due to pulmonary embolism. Measurement of CK isoenzymes proved of great diagnostic value in three patients with sudden circulatory arrest of, at first, unknown cause after successful resuscitation. Acute myocardial infarction was proven by the presence of MB-isoenzyme. In one of these patients an additional BB-isoenzyme was seen, possibly due to concomitant cerebral ischaemia. In all other patients (with angina, after cardioversion, or after major surgical operations) only MM-isoenzyme was detected. MB-CK-isoenzyme was found to be a highly specific, as well as sensitive, indicator of myocardial necrosis. This being a rather difficult method, its use is not justified in the routine diagnosis, but in doubtful instances its value can hardly be overestimated.


[Effectiveness of specially equipped and staffed ambulances in the immediate treatment of cardiac emergencies (author's transl)]. Of 3079 medically justified calls of the special ambulances at Ludwigshafen 70.6% were medical emergencies, of which 1059 were cardiac emergencies, 66.9% of these being myocardial infarctions. One quarter of patients with the typical features of myocardial infarction died in the first few minutes, one third within the first half-hour of onset of symptoms. The use of the special ambulances markedly raised the severity of those cases of infarction admitted to hospital so that, despite appropriate therapeutic measures in the early phase, total mortaltiy of cases of myocardial infarction treated by ambulance and at hospital was 51.1%. On the other hand, very early measures (mechanical and electrical resuscitation, intravenous infusion) can be of decisive importance in patients who would otherwise die as a result of arrhythmias. Long-term survival had been achieved in 29% before use of the special ambulance, in 46.8% after its introduction. No correspondingly good results through very early treatment were achieved after attempted suicide: the death rate was 2.6% both before and after introduction of the special ambulances (470 and 497 cases, respectively).


[Obliterative disease of liver veins: clinical, diagnostic, therapeutic and pathogenetic aspects of Budd-Chiari syndrome (author's transl)]. Both external and internal factors have to be thought of as causes of obliterative disease of liver veins. Pathogenetically, thrombosis or proliferative vascular disease (endophlebitis hepatica obliterans) may affect either large veins (with occlusion of the ostia) or small ones. Veno-occlusive disease, caused by pyrrolizidine derivatives, is a special form. Injection of contrast medium to the inferior vena cava and (or) the liver veins is the most important diagnostic method. In two personal cases there was secondary occlusion of the liver veins as a result of IVC thrombosis, while in a third case there was non-thrombotic proliferative disease of the wall of the small hepatic veins, closely similar to veno-occlusive disease. In a 19-year-old girl in whom thrombosis of the vena cava (with signs of liver-vein occlusion) occurred after pregnancy, revascularisation was achieved by fibrinolytic treatment.


[Intoxication after use of salt as an emetic (author's transl)]. Salt is generally contraindicated as an emetic in toxicological emergency situations. It can only be recommended when its rapid disappearance from the stomach can be guaranteed in the case of lack of vomiting. Less than 1 g salt per kg body weight may be lethal. The danger of sodium chloride becomes apparent from two severe cases of intoxication in children one of which was fatal. One of the two children was given salt as an emetic.


Induction of a coupling defect in rats during inhibition of tyrosine dehalogenase. We have reported earlier that administration of 3-nitro-L-tyrosine (MNT), 8 mM in drinking water, to rats receiving a low iodine diet (LID) results in greater TSH secretion, larger goiters, and more rapid uptake and release of radioiodine than LID alone, and ultimately may produce hypothyroidism. These findings have been confirmed, and hypothyroidism documented by demonstrating depressed levels of hepatic mitochondrial alpha-glycerophosphate dehydrogenase. Also, prolonged treatment with MNT + LID produced a depression in labeled iodothyronine (ITh) synthesis, as judged by chromatographic analysis of thyroid digests from rats killed 4 or 24 hours after ip injection of radioiodine, or two weeks after adding radioiodine to drinking fluid. Low thyroidal ITh levels were accompanied by low levels of ITh in serum, despite the presence of various other labeled organic iodine compounds. Cessation of MNT treatment, or ip injection of small amounts (0.5-1.0 mug) of Na 127I together with radioiodine 4 h before sacrifice reversed the defect, and large amounts of ITh were found in both thyroid and serum. Labeled thyroprotein from MNT-treated rats showed increased susceptibility to disaggregation during freezing at pH 8.5; this abnormality was also reversed by stable iodine treatment. In glands labeled with radioiodine 24 h before sacrifice, stable iodine injection 20 h later was followed by increased thyroidal ITh. It is concluded that profound iodine deficiency, induced by MNT + LID, can lead to diminished ITh synthesis, or a "coupling defect". The results provide an explanation for the finding of low thyroidal ITh in patients with hereditary deficiency of tyrosine dehalogenase. The findings confirm an important role for iodine supply in ITh synthesis and thyroglobulin stability, and suggest that rats treated with MNT + LID provide a model for study of the effects of extreme iodine deficiency.


Effect of neuraminidase treatment on the biological activity of highly purified ovine FSH and LH in hypophysectomized immature male and female rats. Highly purified ovine FSH and LH were treated with neuraminidase to remove sialic acid and the desialylated derivatives were examined for biological activity in hypophysectomized immature male and female rats. The male rats were hypophysectomized at 22 days of age and beginning on day 25 were injected sc twice daily for 4 days with native or neuraminidase-treated FSH (total dose, 15 or 60 mug) or LH (12 mug). The ventral prostates, seminal vesicles, and testes were then removed and weighed, and serum testosterone levels were measured by radioimmunoassay. The female rats were hypophysectomized on day 28 and beginning on day 35 were injected sc twice daily for 4 days with native or neuraminidase-treated FSH (8 mug) or saline. On the morning of day 39, the rats were given an ovulating dose of gonadotropin (8 mug native or neuraminidase-treated FSH, or 1.28 mug native or neuraminidase-treated LH) or 1.0 ml saline iv via tail vein. Twenty-four hours later ova were counted in the oviducts the ovaries were weighed, and serum levels of progesterone and 20alpha-dihydroprogesterone were determined by radioimmunoassay. Treatment of ovine LH with neuraminidase did not diminish the ability of this hormone to increase prostate and testes weights and serum testosterone levels. Desialylation also did not decrease the ability of LH to induce ovulation. Although native ovine FSH significantly increased the weights of the ventral prostate, seminal vesicles, and testes, and elevated plasma testosterone levels, the desialylated derivative was essentially inactive. Neuraminidase treatment also eliminated the ability of ovine FSH to increase ovarian weight, to induce ovulation, and to elevate serum progesterone and 20alpha-dihydroprogesterone. These results indicate that the LH-like activity of ovine FSH is an intrinsic property of the FSH molecule.


Testosterone and dihydrotestosterone concentrations in plasma, epididymal tissues, and seminal fluid of adult rats. The concentration of testosterone (T) and 5alpha-dihydrotestosterone (DHT) in plasma, seminal fluid, and epididymis of male Wistar rats have been determined by radioimmunoassay. The DHT/T ratio rose from 0.1 in blood to 1 in testicular fluid and 4 in epididymal fluid. This ratio is 2 in tissues of "caput" epididymis and 1 in "cauda" epididymis. These data indicate a transformation of T to DHT in the epididymal tissue, particularly at the "caput" level.


Influence of reserpine on high calcium perfusion of thyro-parathyroid glands in dogs. Perfusion of the thyro-parathyroid glands in the dog with calcium-rich blood led to a significant decrease (P less than 0.001) in the systemic plasma calcium level. On the other hand, injection of reserpine (0.2 mg/kg ip) 16 and 2 h prior to infusion of hypercalcemic blood into thyro-parathyroid glands did not alter the systemic plasma calcium level significantly (P less than 0.05). This result provides evidence that reserpine blocks the release of hypocalcemic factor from the thyro-parathyroid glands in response to perfusion of high concentrations of calcium.


Identification of two 5alpha-reduced pregnanes as major metabolites of progesterone in immature rat ovaries (1000 x g supernatant) in vitro. Incubation of the 1000 x g supernatant obtained from 23-day-old rat ovarian homogenate with labeled progesterone resulted in the production of 3 major metabolites; 5alpha-androstane-3alpha,17beta-diol, and two 5alpha-reduced pregnanes that were identified as 3alpha-hydroxy-5alpha-pregnan-20-one and 3alpha,17alpha-dihydroxy-5alpha-pregnan-20-one. The 3alpha,17alpha-dihydroxy-5alpha-pregnan-20-one has not been hitherto isolated from mammalian ovaries. The steroids were identified by their mobilities on thin-layer and gas-liquid chromatography, by mass spectroscopy, derivative formation and by recrystallization to constant specific activity. In another experiment, incubation of the 1000 x g supernatant from 23-day-old rat ovaries with 3alpha-hydroxy-5alpha-pregnan-20-one as substrate resulted in the production of 5alpha-androstane-3alpha,17beta-diol. It is suggested that 5alpha-androstane-3alpha,17beta-diol is produced in immature rat ovaries by a pathway in which the identified 5alpha-reduced pregnanes serve as intermediates.


Relative effectiveness of magnesium and calcium on the secretion and biosynthesis of parathyroid hormone in vitro. The effects of varying extracellular concentrations of calcium and magnesium were compared on the release of 3H-labeled and immunoreactive parathyroid hormone (PTH) by bovine parathyroid slices incubated for 4 h in vitro in media containing [3H]leucine. The quantity of [3H]PTH or immunoreactive PTH released was determined by polyacrylamide gel electrophoresis and by radioimmunoassay, respectively. Biosynthesis of PTH in response to extracellular concentrations of magnesium and calcium was evaluated by radioimmunoassay of amounts of proparathyroid hormone (PROPTH) in extracts of the parathyroid tissues and by electrophoretic analyses of [3H]ProPTH synthesized during a 35 min pulse-labeling of parathyroid slices. Rates of hormone release were found to vary in response to changes in concentrations of either calcium or magnesium. The effects, however, of these two cations on hormone release, although additive, were not equipotent. On a molar basis, approximately 2.5 times as much magnesium as calcium was required to produce equivalent secretory responses. Effects of the cations on hormone biosynthesis wer small; magnesium had no effect and calcium altered synthesis by approximately 15%. Neither cation affected conversion of ProPTH to PTH. The results agree with those reported previously from studies of the effects on hormone secretion of calcium and magnesium in vivo and serve to emphasize that, within normal physiological concentrations of these two cations, calcium is the predominant regulator of PTH secretion and synthesis.


Growth hormone during catch-up growth and failure of catch-up growth in rats. Male Long-Evans rats were fasted or given cortisone injections beginning at 37 days of age in order to produce growth retardation. They were then allowed to recover for periods of up to 28 days. GH concentration was measured in trunk blood plasma of rats decapitated after minimal stress. During the recovery period there was a significant increase in plasma GH in both experimental groups. Organ weight/body weight ratios for liver and heart, protein/tissue, DNA/tissue, and DNA/protein ratios of liver, heart, and skeletal muscle were found to remain normal or to return to normal values during the recovery periods in both experimental groups. DNA content was reduced in both liver and heart at 14 days. At later recovery periods no significant differences from controls were observed. The findings indicate that increased GH release is common to the growth recovery period after both fasting or cortisone treatment. GH concentration in plasma does not correlate with presence or absence of catch-up growth. The organ weight/body weight ratios correlate with previous findings showing prompt return to normal proportions of body weight to tail length in both the cortisone and fast models irrespective of presence or absence of catch-up growth. The results indicate that failure of catch-up growth after cortisone treatment is not the result of decreased pituitary GH secretion. It is probable that multiple factors working in concert are responsible for recovery after transient growth retardation.


Sites of clearance of endogenous parathyroid hormone in the vitamin D-deficient dog. The sites of clearance of endogenous parathyroid hormone (PTH) were studied in dogs who developed secondary hyperparathyroidism on a vitamin D-deficient diet. Simultaneous blood samples were obtained from the femoral artery, and the hepatic, renal, portal, and femoral veins. Radioimmunoassay of canine immunoreactive PTH (iPTH) in a heterologous bovine PTH (bPTH) system indicated that the kidney and liver extracted 49% and 46%, respectively, of the iPTH circulating through these organs. Characterization of the circulating iPTH in these animals was carried out by gel filtration and radioimmunoassay of the eluant fractions utilizing specific amino- and carboxyl-terminal antisera. The hormone in the peripheral circulation co-eluted with [125I]-iodobPTH and no fragments of iPTH were detected. Immunochemical differences between bovine and canine PTH were detected in the carboxyl-terminal region of the molecule. The results indicate that endogenous PTH in the vitamin D-deficient, hypocalcemic dog is cleared by the kidney and liver and that the predominant form of the hormone in this animal is similar to glandular bPTH 1-84.


Maintenance of separated rat pituitary mammotrophs in cell culture. Dispersed female rat anterior pituitary cells were cultured in Medium 199 containing 20% fetal calf serum for 30 days. Prolactin levels in the culture medium remained relatively constant during this time, ranging from 30-40 ng/10,000 cells seeded/day. The total quantity of prolactin released into the medium was 10-15 times that originally contained in the cells. Morphological integrity of the mammotrophs was maintained. Using velocity sedimentation at unit gravity, cells from untreated, overiectomized or estrogen-primed animals were separted into several fractions, and subsequently cultured for 14 days. Not all mammotrophs secreted the same quantity of hormone during this time. The data suggest that the pituitary of the female rat is composed of a heterogeneous population of mammotrophs, and that their capacity to secrete prolactin in vitro may, in part, be reflected by the previous physiological status of the animal.


Glucagon-mediated stimulation of (32P) orthophosphate and (14C) serine incorporation into phosphatidylserine in cardiac muscle slices. Glucagon stimulated the incorporation of Na2H32PO4 and L-(14C)serine into phosphatidylserine in heart muscle slices. The increase above control was about 2-fold at ten minutes and 6-fold at thirty minutes for (32P) and 12-fold as early as three minutes for (14C)serine. Although a smaller, but significant, incorporation of (32P) into phosphatidylethanolamine was also observed, glucagon did not stimulate the incorporation of (14C)serine into phosphatidylethanolamine. Glucagon did not significantly augment the incorporation of either tracer into phosphatidylcholine, lysophosphatidylcholine, phosphatidylinositol, cardiolipin, phosphatidic acid, or sphingomyelin. Dibutyryl cyclic 3',5'-AMP did not increase the incorporation of (32P) or (14C)serine into phosphatidylserine. Since phosphatidylserine appears to serve a critical role in coupling the glucagon receptor to the catalytic moiety of adenylate cyclase, the data suggest that the hormone may initially increase the amount of its own coupler.


The interaction of dopaminergic and serotonergic drugs on plasma prolactin in ovariectomized, estrogen-treated rats. Pharmacologic agents that either block or stimulate dopaminergic or block serotonergic receptors were administered intraarterially alone or in combination to ovariectomized, estrogen-treated rats. Prolactin was estimated by radioimmunoassay of plasma collected by serial blood sampling from chronic aortic catheters. Apomorphine (10 mg/kg) depressed, whereas Pimozide (50 and 250 mug/kg) and Methiothepin (50, 500 and 5000 mug/kg) markedly increased prolactin levels. Apomorphine completely blocked the increases induced by the lowest does of Pimozide and Methiothepin, as well as the initial increases at the higher doses. However, at later time intervals after large doses of Pimozide and Methiothepin in apomorphine-pretreated rats there was a marked rebound of prolactin which was greater than the level induced by each drug alone. The serotonin antagonists SQ 10,631 and methysergide reduced the Pimozide-induced elevation in prolactin levels, suggesting that part of the increase in prolactin following dopaminergic blockage may result from the action of a tonically stimulatory serotonin system that is normally masked by the intense tonic inhibition by the dopaminergic system.


The purification and characterization of ovine placental lactogen. Ovine placental lactogen (oPL), has been purified approximately 1,000-fold from sheep cotyledons using conventional protein purification procedures. Radioreceptor assays using rabbit liver particulate fractions for growth hormone (RRA-GH) and using rabbit mammary gland particulate fractions for prolactin (RRA-PRL) were employed to monitor the hormonal activities. The molecular weight of oPL is approximately 22,000 as determined by gel filtration on Sephadex G-100, and its isoelectric point as determined by isoelectric focusing is 8.8. In the two RRA's, the displacement curve of oPL is parallel to bovine growth hormone (bGH) and ovine prolactin (oPRL) standards and the ratio of GH-activity to PRL-activity of oPL is 1:2. In a body weight gain assay using hypophysectomized rats, oPL has a growth-promoting potency of 1.3 U/mg. In rabbit mammary explants, oPL stimulates casein synthesis. In a receptor assay for growth hormone using human liver, oPL and hGH are equipotent in competing for receptor sites, suggesting that oPL and hGH have common structural features that are lacking in other non-primate hormones.


Conversion of L-thyroxine to triiodo-L-thyronine and biological activity of L-thyroxine, as measured by changes in growth hormone. Pituitary growth hormone (GH) content and plasma GH response to pentobarbital (PB) of severely hypothyroid rats are markedly decreased compared with euthyroid rats. Both pituitary and plasma GH may be increased by single ip injections of physiologic doses of thyroxine (T4) (1.75 mug/100g BW) or triiodothyronine (T3) (0.2 mug/100g BW). Using this increase in GH of hypothyroid rats to measure the biological effectiveness of single doses of both iodothyronines, we have shown that the administration of 6-propyl-2-thiouracil (PTU) interferes with the activity of T4, but not with that of T3. It is known that deiodination of T4 and of T3 and conversion of T4 to T3 are partially inhibited by PTU. Therefore, it appears that deiodination of T4 is biologically important, whereas deiodination of T3 is not. Data presented here are thus consistent with the hypothesis that conversion of T4 to T3 in vivo plays an important role in the expression of T4 activity.


Antidiuresis produced by injection of renin into the third cerebral ventricle of the dog. The purpose of this study was to determine whether centrally administered renin stimulated vasopressin secretion. Vasopressin was not measured directly, but, instead, changes in urinary water excretion in anesthesized dogs undergoing a water excretion in anesthetized dogs undergoing a water diuresis were used as an index of changes in vasopressin secretion. Intraventricular injection of hog renin in a dose of 0.1 Goldblatt unit produced a marked decrease in urine flow which was associated with a decrease in free water clearance and an increase in urinary osmolatiy with no change in osmolar clearance. Sodium excretion increased significantly but there was no change in potassium excretion. These effects, which closely resemble those resulting from an increase in vasopressin secretion, were prevented by hypophysectomy. The antidiuretic effect clearly resulted from an action of renin in the central nervous system since renin had no effect on urine flow or osmolality when administered intravenously. Intraventricular administration of saralasin acetate, a specific antagonist of angiotensin II, completely blocked the effects of intraventricular renin indicating that these effects were mediated via the formation of angiotensin II. The data therefore indicate that there is an interaction between injected renin, brain angiotensinogen, and converting enzyme resulting in the formation of angiotensin II which stimulates the secretion of vasopressin. Additional studies are required to determine whether the brain renin-angiotensin system plays a physiological role in the regulation of a vasopressin secretion.


Beta adrenergic receptor dysfunction in hypoxic inhibition of insulin release. Insulin secretory responses were measured in air-breathing puppies given epinephrine, and compared with insulin responses during acute hypoxia. In puppies with oxygen deficiency, insulin levels declined, whereas during epinephrine infusion, they remained stable or increased slightly. When glucose was given during phentolamine administration, insulin levels rose more in the epinephrine-treated animals than in the hypoxic animals, despite similar blood glucose levels. Theophylline-induced insulin release was increased by epinephrine, but inhibited by hypoxia. When the beta adrenergic blocking agent, propranolol, was given with the epinephrine, the insulin response to theophylline was markedly reduced and similar to that observed during hypoxia. Control studies with propranolol showed no effect of this agent on glucose-induced insulin release. Isoproterenol infusion caused elevated insulin levels during air ventilation but this response was suppressed by hypoxia. From these data, we have concluded that the difference between hypoxia and epinephrine can be explained by a reduced ability of catecholamines to stimulate the beta adrenergic receptor during hypoxia. We hypothesize that this effect leads to an unmodulated alpha adrenergic inhibition of insulin release.


Effects of morphine and methadone on serum testosterone and luteinizing hormone levels and on the secondary sex organs of the male rat. The effects of morphine and methadone on the endocrine control of the male rat's sexual function were examined. The results indicate that these narcotics markedly reduce the structural and functional integrity of the secondary sex organs by producing a pronounced reduction in serum testosterone levels. Serum levels of luteinizing hormone (LH) were not detectable in narcotic-treated animals, whereas serum levels of the follicle stimulating hormone (FSH) were unaltered. On the basis of these observations, it seems reasonable to conclude that the narcotics inhibit the secretion of LH, by an action either in the hypothalamus (e.g., suppression of LH-releasing hormone) or directly in the pituitary gland, which leads to a reduction in serum testosterone levels and a subsequent reduction in the wet-tissue weight and secretory activity of the secondary sex organs.


Changes in blood flow to bones during the hypocalcemic and hypercalcemic phases of the response to parathyroid hormone. Labeled microspheres were used to measure blood flow to the leg bones of the laying hen at 0, 3, and 30 min after iv injection of parathyroid hormone (PTH) (Wilson) or the synthetic 1 to 34 fragment of bovine parathyroid hormone (PTH 1-34). At 3 min, which corresponds to the hypocalcemic phase of the PTH response, blood flow to the combined femur, tibia, and metatarus was significantly reduced by PTH (Wilson) relative to 0 time and to carrier-injected controls. At 30 min, i.e., the time of maximum hypercalcemia in the hen, blood flow to these bones was significantly increased relative to 0 time. The results obtained with PTH 1-34 were similar, except that the decrease at 3 min was only significant in comparison with the controls injected with inactivated hormone. Femoral blood flow and the venous minus arterial calcium gradients across the femur were positively correlated, irrespective of sampling time (0 or 30 min) or type of injection (PTH [Wilson] or carrier). Taken together, these results suggest that there is a relationship between calcium mobilization from bone and the rate of osseous blood flow. Other organs which showed significant changes in blood flow after PTH (Wilson) were the adrenals, thyroids, and shell gland; the cerebellum, parathyroids, heart, spleen, liver, pancreas, duodenum, magnum, isthmus, and kidneys were not affected.


Thyroid hormone regulation of extrathyroidal iodoproteins. The effect of thyroid status on plasma and tissue levels of labeled nonextractable iodine (NEI) derived from the metabolism of radioiodothyronines was examined in the rat. Concentrations of radioiodoprotein were substantially elevated in plasma, kidney, and liver in thyroidectomized animals 72 h postinjection of [125I]triiodothytonine ([125I]T3). Similarly, total rat concentrations of radioactive NEI were increased (52%) 72 h after injection of [125I]T3. NE125I concentrations from [125I]T3 in plasma, kidney, and liver were diminished progressively in thyroidectomized animals maintained on increasing doses of thyroxine replacement, demonstrating that iodoprotein levels were inversely related to thyroid state. The plasma disappearance rate of radioiodoprotein from [125I]T3 was markedly slowed in hypothyroid animals and accelerated in intact controls rendered hyperthyroid with daily injections of T4, 8 mug/100 g BW. Propylthiouracil (PTU) treatment of thyroidectomized rats maintained on T4, 2 mug/100 g BW per day resulted in increased NE125I from [125E]T3 in plasma, kidney, and liver. The results of the foregoing investigations suggest that thyroid hormone regulates levels of iodothyronine-derived iodoproteins by influencing the rate of degradation of iodoproteins. Moreover, the observed elevation of iodoprotein levels in T4-maintained thyroidectomized animals after PTU administration appears consistent with the modification of thyroid status due to the peripheral antithyroxine effect of PTU.


Radioimmunoassay of serum concentrations of melatonin in sheep exposed to different lighting regimens. A specific and sensitive double-antibody radioimmunoassay for melatonin (N-acetyl-5-methoxytryptamine) has been developed utilizing rabbit antisera to a bovine serum albumin conjugate of N-succinyl-5-methoxytryptamine and utilizing N-3-(4-hydroxyphenl)-propionyl-5-methoxytryptamine for radioiodination. The least detectable concentration of melatonin standard was 10 pmolar (2.3 pg/tube) with 50% inhibition resultinhibition curves obtained with increasing quantities of melatonin or increasing quantities of chloroform extracts of ovine sera were parallel. The immunoreactivity found in ovine sera c-migrated with [3H]melatonin on silica gel G when developed with chloroform:methanol (9:1). N-Acetylserotonin, 5-methoxytryptamine, serotonin, tryptophan, 6-hydroxymelatonin, 6-methoxytetrahydroharmalan, and several other indole and beta-carboline compounds do not influence the estimation of melatonin in the radioimmunoassay. Concentrations of melatonin could be accurately determined when 31 to 1000 pg were added to 1 ml ovine serum. Serum samples with melatonin concentrations of 1000 pg/ml, 500 pg/ml and 75 pg/ml had intra-assay coefficients of variation of 9.1%, 8.6%, and 17.4%, respectively. The respective inter-assay coefficients of variation were 22.7%, 18.1%, and 37.1%. Ewes exposed to a 12 h light: 12 h dark lighting regimen demonstrated a circadian rhythm in serum concentrations of melatonin. Concentrations ranged from 10-30 pg/ml during periods of light to 100-300 pg/ml during periods of dark. During exposure to continuous light, the circadian rhythm was abolished and concentrations of melatonin were maintained at 10-50 pg/ml. When exposed to conditions of continuous dark the circadian rhythm persisted. A precipitous drop in serum concentrations of melatonin resulted when ewes experiencing peak melatonin concentrations were exposed to light. Concentrations returned to peak levels when the lights were turned off 3.5 h later.


Effect of luteinizing hormone on respiration of the preovulatory cumulus oophorus of the rat. The effect of luteinizing hormone (LH) on the respiration of isolated cumulus cell complexes (the oocyte surrounded by cumulus granulosa cells) obtained from immature Sprague-Dawley rats injected with 10 IU pregnant mare's serum gonadotropin on day 30 was investigated. The cell complexes were isolated from preovulatory follicles of rats killed at specific time intervals on the day preceding ovulation, i.e., on day 32. The samples were incubated in Eagle's tissue culture medium. Oxygen uptake was recorded either with the Cartesian diver technique or with a recently described microspectrophotometric technique using hemoglobin as an indicator. Exposure to exogenous bovine LH in vitro or in vivo or to endogenous LH, i.e., the preovulatory LH-surge, resulted in a marked decrease in respiratory activity of the cumulus cell complex, as revealed by both techniques. The cumuli exposed to LH showed an oxygen uptake of approximately 40-65% of the control cumuli. The results suggest that LH has a direct effect on this cell complex resulting in a decreased oxidative metabolism.


Effects of mating on serum LH, FSH, and prolactin and accessory tissue weight in male rats. The literature reveals contradictory data regarding whether or not male release LH acutely following coitus. Since it has been shown that repeated mating increases accessory sex organ weight, suggesting induction of gonadotrophin release, it seemed of interest to reinvestigate the issue. Male rats were divided into two groups: the mated group was provided with frequent mating ("experiencing") trials; the unmated group served as cage controls which received no sexual contact throughout the entire experiment. All rats were provided with chronic jugular cannulae. One set of serum samples was taken from each rat at 85 days, the second set at 115 days. On the two evening when sampling took place the mated rats were subdivided into three groups: "chamber" (placed in mating arena alone), "mount" (allowed two to five mounts, but no intromission), and "ejaculation" (mated through entire copulatory sequence). Serum sampling took place every 15 min during the hour following completion of mating (defined in the "ejaculation" group). On the day following the second sampling, autopsies were performed, and terminal serum samples were taken. LH, FSH, and prolactin were measured by radioimmunoassay. There were no significant differences in any hormone among the groups during the hour following mating nor in the terminal sample. However, the mated rats showed significantly greater seminal vesicle and ventral prostate weights than the cage controls at autopsy. Several hypotheses are offered to account for the latter findings.


An acrylamide gel recombination assay for glycoprotein hormone subunits. A specific and sensitive assay for recombination of the subunits of hCG and other glycoprotein hormones (LH, FSH, and TSH) is presented. The assay depends on the ability of sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) to separate clearly the subunits from native hormone. Iodinated subunit is incubated with its identical unlabeled subunit and complementary subunit. Quantitative displacement of the radioactivity to the native hormone position as followed by gel slicing is used as a direct index of recombination. The assay is reproducible, sensitive to less than one picomole of hormone formed, and offers a simple means of evaluating recombination of glycoprotein chains as distinct from biological activity.


Further studies on the hypercalcemic effect of acute calcitonin deficiency in rats. Studies were carried out to determine the origin of the immediate increase in plasma calcium following acute calcitonin deficiency in mature rats. Animals were pre-labelled with 45calcium 24 hours and 4 weeks before thyroparathyroidectomy (TPTX) and 1 hour, 24 hours, 2 and 4 weeks before nephrectomy and TPTX and bled serially over the following 3 hours. In each study the final average weight of the rats was over 300 g. Plasma calcium increased after TPTX. In rats labelled with 45calcium 1 and 24 hours previously, the rise was to small to alter the specific activity of calcium although radiocalcium was unchanged. In contrast, in animals pre-labelled with 45calcium 2 and 4 weeks before TPTX, the increase in stable calcium was associated with a parallel increase in radiocalcium. Consequently, the specific activity of plasma calcium did not differ appreciably from that of controls. These findings confirm the theory that in mature unfed rats acute calcitonin deficiency results in an immediate rise in plasma calcium. Since this increase is due mainly to enhanced transport of calcium from deep bone, our observations are in accord with the view that calcitonin decreases plasma calcium primarily by inhibiting calcium transport from "stable" bone.


Blockade of the stress-induced decrease in blood GH by anti-somatostatin serum in rats. To investigate the physiological role of somatostatin in the GH release mechanism, the effect of passive immunization with somatostatin on basal and poststress serum GH levels was examined in rats. Sheep antiserum to somatostatin or normal sheep serum (NSS) was injected iv into adult male rats 5 days, 3 days and 1 day before applying stressful stimuli. Thirty min after the first bleeding under ether, the mean serum GH level increased from 36.3 to 76.1 ng/ml in the antiserum-treated rats and decreased from 63.7 to 16.8 ng/ml in the NSS-treated rats, but the difference for either group was not significant due to the large variation. In another experiment, electroshock was applied to conscious rats. The mean poststress GH level in the rats pre-treated 3 h previously with the antiserum was 34.4 ng/ml, a value significantly higher than the GH level in the rats pretreated with NSS (0.9 ng/ml) or in non-treated rats (0.5 ng/ml). It was concluded that the stress-induced decrease of GH secretion in rats is at least partly due to an increased release of hypothalamic somatostatin and/or immunologically related GH release-inhibiting hormone(s). Persistent variation in serum GH levels after neutralization of endogenous somatostatin by the antiserum suggests that another regulator, probably GH releasing hormone, also plays a role in controlling GH secretion.


Purification and properties of an esterase B4 from human liver. A butyryl esterase, designated B4, has been purified from human liver and some of its properties described. The activity of this enzyme comprises 0.48% of the total butyryl esterase activity found in human liver. Esterase B4 has been distinguished from other butyryl esterases by its preference for the esters of the fluorogenic compounds 4-methyl umbilliferone and fluorescein over naphthyl esters as substrates. Other distinguishing features of this esterase include a relatively high pI (pH 8.7) A monomeric structure of low molecular weight (20 000) and high solubility in solutions of ammonium sulphate.


Interactions of KB-cell glycoproteins with an adenovirus capsid protein. Glycoprotein material extracted from human KB cells with a flurocarbon, trichlorotrifluoroethane (Arklone P), into a waste-soluble fraction binds to fibre, a structural protein of the adenovirus type-5 capsid. The fibre-binding glycoprotein(s) were purified by ion-exchange chromatography on DEAE-Sephadex and affinity chromatography on a fibre-Sepharose support. The purification procedure also includes a trypsinization step which eliminates the bulk of contaminating KB cell proteins present in the aqueous fraction without appreciably affecting the activity of the fibre-binding glycoprotein(s). Some comparison is made of the membrane-bound receptors for adenovirus and the water-soluble fibre-binding glycoprotein(s).


31P nuclear-magnetic-resonance studies on the developing embryos of Xenopus laevis. The concentrations of nucleoside triphosphate, inorganic phosphate and the yolk proteins, phosvitin and lipovitellin, have been monitored in living embryos of Xenopus laevis by 31P nuclear magnetic resonance (NMR) spectroscopy. The nucleoside triphosphate levels remain relatively constant at about 3.5-4.5 nmol/embryo at least until the 'spontaneous movement' stage of development. By the swimming tadpole stage an inorganic phosphate resonance representing about 30 nmol/embryo becomes evident in the NMR spectrum. Computer manipulation also shows such a resonance, although smaller, to be present at a somewhat earlier developmental stage; these findings are confirmed biochemically. The major contribution to the NMR spectrum of oocytes, unfertilized eggs and early embryos is the yolk phosphoprotein resonance. On isolation of the yolk from the embryos it is possible to quantify  the contribution to the NMR spectrum from the lipid-phosphate and protein-phosphate moieties of the yolk proteins. During development, as the yolk is used up, it is found that the protein-phosphate resonance disappears at a greater rate than the lipid-phosphate peak. The total phosphorus content of the embryo (approximately 200 nmol/embryo) is shown biochemically to remain constant during development; however, the total amount of phosphorus observed by NMR decreases by about 40% during development. From the resonance positions of their alpha, beta and gamma phosphate groups it is deduced that the nucleoside triphosphate molecules are liganded in vivo to a divalent cation which is not manganese, but could be either magnesium or calcium. From the position of the inorganic phosphate resonance it is deduced that the internal pH of embryos where this resonance is evident is 6.8 +/- 0.2.


Subcellular localization of a lesion in protein synthesis in rabbit reticulocytes incubated at elevated temperatures. When rabbit reticulocytes are incubated at 43-45 degrees C their rate of protein synthesis rapidly decreases, compared to a contol 37 degrees C incubation. Lysates prepared from cells incubated at this supra-optimal temperature have an equally decreased capacity for endogenous, but not poly(uridylic acid)-directed, protein synthesis. Subcellular fractionation traced the lesion to the crude ribosomal pellet, 0.5 M KCl ribosomal wash and postribosomal supernatant of the temperature-shocked cells. Preparation of purified ribosomal subparticles showed, however, that they were as active as the control in protein synthesis. In this paper we present evidence that the decreased activity of the heated lysate, 0.5 mM KCl wash and postribosomal supernatant is due to an inhibitor and can be overcome by the addition of 0.5 M KCl or supernatant from control cells. The results are discussed in terms of the inactivation of a component, essential for initiation of endogenous protein synthesis, which is probably partitioned between ribosomes and supernatant. We also suggest that the decreased protein synthetic activity of the heated cells may be related to their decreased synthesis of haem.


A mutant strain of Chlamydomonas reinhardi exhibiting altered ribulosebisphosphate carboxylase. A mutant, ac i72, of Chlamydomonas reinhardi possessing an altered ribulosebisphosphate carboxylase and unable to grow on minimal medium has been isolated and characterized. Comparison of ribulosebisphosphate carboxylase purified from both wild type and ac i72 strains is given. The enzyme from ac i72 shows alterations in several characteristics: (a) the specific activity is reduced to 35% that of wild type, (b) the V for both substrates is reduced 3-6 fold, (c) the Mg2+ requirement for maximal activity is 3 times greater, (d) the inhibitory effect of Cl- is greater, and (e) the isoelectric point is changed (6.0 for wild type and 5.8 for ac i72). However, the ribulosebisphosphate carboxylase from ac i72 is identical to that from wild type with respect to pH requirement, temperature sensitivity, subunit structure, and sedimentation characteristic. Other photosynthetic properties of wild type and ac i72 cells were also compared. CO2 fixation in ac i72 in vivo is reduced proportionally to the reduction in activity of the enzyme, but the level of O2 evolution is the same as in wild-type cells. Photosynthetic electron transport, 70-S ribosome content, and chlorophyll content are unaltered in ac i72. The chloroplast ultrastructure of ac i72 cells is distinctly different from that of wild-type cells. The inheritance of the mutation is Mendelian.


New assay for pregnenolone and progesterone 17alpha- hydroxylase based on the specific substitution of a tritium situated on carbon 17. A new assay for the measurement of steroid 17alpha-hydroxylase activity in beef adrenals is described. This method is based on the biochemical mechanism of the enzymic reaction, i.e. the direct and stereospecific substitution of the proton located on the hydroxylated position. Progesterone or pregnenolone specifically labelled on the 17 position are solubilized in the incubation mixture with the help of Tween 80 and incubated under optimal conditions. The tritium enzymically released from the substrate is found in the medium as a molecule of water which is then distilled under reduced pressure and counted by liquid scintillation. The results obtained with this new method are comparable with those obtained with a conventional method using a 14C-labelled substrate.


Cortisol modification of HeLa 65 alkaline phosphatase. Decreased phosphate content of the induced enzyme. Alkaline phosphatase activity of HeLa cells is increased 5-20-fold during growth in medium with cortisol. The increase in enzyme activity is due to an enhanced catalytic efficiency rather than an increase in alkaline phosphatase protein in induced cells. In the present study the chemical composition of control and induced forms of alkaline phosphatase were investigated to determine the enzyme modification that may be responsible for the increased catalytic activity. HeLa alkaline phosphatase is a phosphoprotein and the induced form of the enzyme has approximately one-half of the phosphate residues associated with control enzyme. The decrease in phosphate residues of the enzyme apparently alters its catalytic activity. Other chemical components of purified alkaline phosphatase from control and induced cells are similar; these include sialic acid, hexosamine and sulfhydryl residues.


The protein-mediated transfer of phosphatidylcholine between membranes. The effect of membrane lipid composition and ionic composition of the medium. The phosphatidylcholine exchange protein from bovine liver catalyzes the transfer of phosphatidylcholine between rat liver mitochondria and sonicated liposomes. The effect of changes in the liposomal lipid composition and ionic composition of the medium on the transfer have been determined. In addition, it has been determined how these changes affected the electrophoretic mobility i.e. the surface charge of the membrane particles involved. Transfer was inhibited by the incorporation of negatively charged phosphatidic acid, phosphatidylserine, phosphatidylglycerol and phosphatidylinositol into the phosphatidylcholine-containing vesicles; zwitterionic phosphatidyl-ethanolamine had much less of an inhibitory effect while positively charged stearylamine stimulated. The cation Mg2+ and, to a lesser extent, K+ overcame the inhibitory effect exerted by phosphatidic acid, in that concentration range where these ions neutralized the negative surface charge most effectively. Under conditions where Mg2+ and K+ affected the membrane surface charge relatively little inhibition was observed. In measuring the protein-mediated transfer between a monolayer and vesicles consisting of only phosphatidylcholine, cations inhibited the transfer in the order La3+ greater than Mg2+ larger than or equal to Ca2+ greater than K+ = Na+. Inhibition was not related to the ionic strength, and very likely reflects an interference of these cations with an electrostatic interaction between the exchange protein and the polar head group of phosphatidylcholine.


Purification and characterization of dopamine beta-hydroxylase from bovine adrenal medulla. A new purification procedure that permits large-scale purification of dopamine beta-hydroxylase from bovine adrenal medulla was developed. Whole adrenal medullas were extracted with 0.1% Triton X-100, and the enzyme was purified by precipitation with polyethylene glycol, chromatography on DEAE-cellulose, and adsorption to concanavalin A linked to agarose. The yield of protein and the specific activity were high compared with previously published methods. The enzyme appeared essentially homogenous by the criteria of polyacrylamide gel electrophoresis in the presence or absence of dodecylsulfate, and sedimentation velocity analysis. The purified protein was subjected to amino acid and carbohydrate analyses, and the results were compared with previously published data. We found about 3 mol of copper per mol of protein (tetramer of 290000 daltons). No free sulfhydryl groups could be found. Analysis for NH2-terminal amino acids with [14C]dansyl chloride revealed 2 residues of alanine and 2 residues of serine per tetramer. We found the NH2-terminal amino acid of chromogranin A to be leucine. The results of our analysis for amino acid composition and NH2-terminal amino acids do not support the suggestion that dopamine beta-hydroxylase and chromogranin A contain identical peptide chains.


Chemical modification as a probe of the topography and reactivity of horse-spleen apoferritin. In apoferritin, but not in ferritin, 1.0 +/- 0.1 cysteine residue per subunit can be modified. In ferritin 3.3 +/- 0.3 lysine residues and 7.1 +/- 0.7 carboxyl groups per subunit can be modified, whilst the corresponding values for apoferritin are 4.4 +/- 0.4 lysine residues and 11.0 +/- 0.4 carboxyl groups per subunit. Modification of lysine residues which maleic anhydride and carboxyl groups with glycineamide in apoferritin which has been dissociated and denatured in guanidine hydrochloride leads to the introduction of 9.1 +/- 0.5 maleyl groups per subunit and 22.0 +/- 0.9 glycineamide residues per subunit. Whereas unmodified apoferritin subunit can be reassociated from guanidine hydrochloride to apoferritin monomer, the ability of maleylated apoferritin to reassociate is impaired. Apoferritin in which all the carboxyl groups have been blocked with glycineamide cannot be reassociated to apoferritin and exists in solution as stable subunits. The modification of one cysteine residue per subunit, of 3 or 4 lysine residues per subunit or of 7 carboxyl groups per subunit has no effect on the catalytic activity of apoferritin. In contrast the modification of 11 carboxyl groups per subunit completely abolishes the catalytic properties of the protein. We conclude that one or more carboxyl groups are essential for the catalytic activity of horse spleen apoferritin.


Localization of a prostaglandin F2alpha receptor in bovine Corpus luteum plasma membranes. The distribution of a prostaglandin F2alpha receptor in various subcellular fractions from bovine corpora lutea obtained by differential and gradient centrifugation paralleled very closely the distribution in these fractions of 5'-nucleotidase, a marker enzyme for plasma membranes. The fractions most enriched in the receptor and 5'-nucleotidase were relatively free of mitochondria and lysosomes but were contaminated to some extent by elements of the endoplasmic reticulum. From these results it can be concluded that the prostaglandin F2alpha receptor is localized on the plasma membranes of the corpus luteum cells. A simple method is described for the purification of plasma membranes from bovine corpora lutea by differential centrifugation.


Pyruvate carboxylase: affinity labelling of the magnesium adenosine triphosphate binding site. The 2' , 3'-dialdehyde derivative of ATP (oATP) was prepared by periodate oxidation and on the following criteria was considered to be an effective affinity label.  The magnesium complex of this derivative (Mg-oATP2) was shown to ba linear competitive inhibitor with respect to MgATP2-in both the acetyl-CoA-dependent and -independent activities of the enzyme but was a non-competitive inhibitor with respect to bicarbonate, and an uncompetitive inhibitor with respect to pyruvate.  Mg-oATP was covalently bound to pyruvate carboxylase by reduction using sodium borohydride with concurrent irreversible inactivation of the enzyme...


A DNA polymerase from Ustilago maydis. 1. Purification and properties of the polymerase activity. A DNA polymerase from Ustilago maydis has been purified to apparent homogeneity. The native enzyme possesses a subunit structure consisting of 50000 and 55000-dalton monomers. The apparent sedimentation coefficient of the polymerase activity in the absence of salt is 8.4 S (Mr=180000-200000), that in its presence (0.6 M NaCl or 0.12 M KCl) being 6.3 S (Mr=80000-100000). Low concentrations of EDTA also converted the 8.4-S to a 6.3-S form, whereas magnesium ions catalysed the reverse association. The enzyme has an absolute requirement for both a DNA or RNA template and a DNA primer. For homopolymer templates the primer requirement was satisified by a short complementary oligodeoxynucleotide, but oligoribonucleotides were extremely inefficient primers. With the template-primer poly(dA) X (dT)12, the enzyme added an average of 50 dTMP nucleotides on to each primer molecule, whereas with poly(rA) X (dT)12, this figure was 300. The enzyme also possesses an associated deoxyribonuclease activity. No other DNA polymerase activity was detected in cell-free extracts of U. maydis.


A DNA polymerase from Ustilago maydis. 2. Properties of the associated deoxyribonuclease activity. The polymerase and deoxyribonuclease activities of the purified Ustilago maydis DNA polymerase coeluted from a hydroxyapatite column, cosedimented in sucrose gradients in both the absence and presence of salt, possessed similar thermolabilities and reaction requirements. These observations suggest that both activities are associated with the same enzyme and that the deoxyribonuclease activity is not a contaminant. The initial rate of degradation of native 3'-end-group-labelled DNA was similar to that of a heat-denatured substrate, but the final extent was greater for the former. The enzyme exhibits a high specificity for degradation of DNA in a 3' leads to 5' direction. The degradation of a DNA template was inhibited by the presence of the deoxyribonucleoside triphosphates necessary for simultaneous DNA synthesis, but not that of the newly synthesised DNA. About 50%, 29% and 13% of the purine, cytosine and thymine deoxyribonucleotide residues incorporated by the enzyme into DNA respectively, were subsequently excised when monitored by the resulting conversion of the triphosphate substrates to free monophosphate. The majority of the purine deoxyribonucleoside monophosphates appear after the synthetic phase of the reaction has ceased. In many respects, therefore, the deoxyribonuclease activity of the U. maydis DNA polymerase is similar to the bacteriophage T4-induced enzyme.


Purification and properties of formaldehyde dehydrogenase and formate dehydrogenase from Candida boidinii. Formaldehyde hydrogenase and formate dehydrogenase were purified 130-fold and 19-fold respectively from Candida boidinii grown on methanol. The final enzyme preparations were homogenous as judged by acrylamide gel electrophoresis and by sedimentation in an ultracentrifuge. The molecular weights of the enzymes were determined by sedimentation equilibrium studies and calculated as 80000 and 74000 respectively. Dissociation into subunits was observed by treatment with sodium dodecylsulfate. The molecular weights of the polypeptide chains were estimated to be 40000 and 36000 respectively. The NAD-linked formaldehyde dehydrogenase specifically requires reduced glutathione for activity. Besides formaldehyde only methylglyoxal served as a substrate but no other aldehyde tested. The Km values were found to be 0.25 mM for formaldehyde, 1.2 mM for methylglyoxal, 0.09 mM for NAD and 0.13 mM for glutathione. Evidence is presented which demonstrates that the reaction product of the formaldehyde-dehydrogenase-catalyzed oxidation of formaldehyde is S-formylglutathione rather than formate. The NAD-linked formate dehydrogenase catalyzes specifically the oxidation of formate to carbon dioxide. The Km values were found to be 13 mM for formate and 0.09 mM for NAD.


Characterization of polysomes from Xenopus liver synthesizing vitellogenin and translation of vitellogenin and albumin messenger RNA's in vitro. 1. Conditions are described for the isolation of polysomes from the liver of Xenopus laevis. The method involves homogenization of liver in 0.2 M Tris-HCl pH 8.5, treatment with 2% Triton X-100  and subsequent sucrose density gradient fractionation of polysomes from a 10000 X g supernatant. 2. Vitellogenin synthesis was induced in male Xenopus liver by oestradiol treatment. Polysomes were isolated and vitellogenin-synthesizing polysomes characterized by their association with monospecific 125 I-labelled rabbit anti-vitellogenin antibody and by reaction with rabbit anti-vitellogenin immunoglobulins followed by indirect immunoprecipitation with goat anti-rabbit antibody. 3. Changes in liver polysome content following oestrogen treatment of male Xenopus are correlated with the appearance of vitellogenin synthesis using an organ culture assay. 4. RNA extracted from livers of oestradiol-treated male Xenopus and from purified polysomes is shown to code for the synthesis of vitellogenin-specific immunoprecipitable polypeptides in a rabbit reticulocyte cell-free protein-synthesizing system, a major component having a molecular weight of 210000. Xanopus liver RNA is also shown to code for the synthesis of an albumin-specific immunoprecipitable polypeptide of 74000 molecular-weight which coelectrophoresed with Xenopus albumin.


Renaturation of bacteriophage lambda DNA. Determination of the optimal renaturation conditions using a single-strand-specific DNase and alkaline-sucrose-gradient assay system. Reannealed hybrid molecules of wild-type bacteriophage lambda DNA were prepared in aqueous solutions of formamide at a variety of NaCl concentrations at both room temperature ( 22 degrees C) and 37 degrees C. Treatment of the hybrid DNA molecules with the single-strand-specific nuclease S1 from Aspergillus oryzae followed by alkaline sucrose gradient sedimentation was used to monitor the extent and fidelity of hybridization. The optimal renaturation conditions at room temperature were found to be: 50% formamide, 35-55 mM NaCl and 10 mM Tris-HCl (pH 8.5) at 20-25 mug DNA/ml. Optimal conditions at 37 degrees C were: 32% formamide, 35-55 mM NaCl and 10 mM Tris-HCl (pH 8.5) at 20-25 mug DNA/ml. Under these conditions approximately 85-90% of the input single-stranded DNA (molecular  weight 1.5 X 10(7)) was rendered S1-nuclease-resistant within 8 h at room temperature and 5 h at 37 degrees C. Neither Mg2+ nor spermidine appeared to have an effect on either the extent or fidelity of duplex formation. Experiments performed with excess enzyme and with lambda/lambda imm 434 heteroduplex hybrids suggested that the hybrid that the hybrid DNA molecules formed under optimal conditions contained no, or only short (less than 1%), mismatched regions.


A plasmocytoma ribosome-associated protein kinase which phosphorylates a specific protein of the ribosomal KCl wash. One ribosomal protein kinase activity and 3 soluble protein kinase activities have been identified in plasma cell tumors by DEAE-cellulose chromatography. We have shown phosphorylation in vivo and in vitro of a protein fraction from the ribosomal KCl wash which we have termed 'PPx fraction'. Phosphorylation of this protein fraction has been obtained in vitro with the ribosome-associated protein kinase. We have determined for the ribosomal protein kinase the following characteristics. 1. It is an Mg2+-dependent enzyme that transfers the gamma-phosphate from ATP into phosphoseryl and phosphothreonyl residues of the substrate. 2. It has a wide substrate specificity. Like the soluble protein kinases it catalyses the phosphorylation of several proteins like histone, phosvitin, casein and ribosomal proteins but it differs from the main soluble kinases (I, II) by the fact that it catalyses specifically the phosphorylation at least of one of the ribosomal KCl wash proteins. On dodecylsulfate-polyacrylamide gels this protein has a molecular weight of approximately 90000 and it is released from ribosomes under conditions commonly employed for extraction of initiation factors. 3. The ribosome-associated protein kinase is not stimulated by the addition of cyclic adenosine 3':5'-monophosphate. 4. KCl has no effect, NaCl has a weak effect on the phosphorylation, Mn2+ and Ca2+ are inhibitors. 5. ADP has been found to be a competitive inhibitor. 6. The maximum velocity of the ribosomal protein-kinase-catalysed reaction is 0.65 nmol of 32P incorporated in the KCl wash protein per min and per mg protein. 7. The apparent Km for the ribosomal KCl wash protein as substrate is 0.71 mg/ml and the Km for ATP is 94 muM. 8. The molecular weight of the ribosomal protein kinase, estimated by electrophoresis in polyacrylamide-dodecylsulfate gels, is 60000 and corresponds probably to a catalytic subunit.