Binding of human chorionic gonadotropin by rat ovarian slices: dependence on the functional state of the ovary (38571).

Ovarian receptors specific for hCG-LH can be induced by PMSG-ovine LH priming.  The marked increase in binding activity was accompanied by a rise in ovarian progesterone concentration and preceded by ovarian weight increase. The high binding activity of heavily luteinized ovaries was followed by a decline to prepriming levels during luteolysis.  The ovarian progesterone content was decreased simultaneously and no change in ovarian weight was evident.  The data demonstrate that ovarian hCG binding activity undergoes variation which depends on the functional state of ovaries.


Influence of altering total body sodium on angiotensin I systemic converting activity (38573)

Studies were carried out in the dog to determine the extent to which alterations in total body sodium influence the systemic conversion of angiotensin 1.  When the animals were maintained on a high salt diet and DOCA, conversion of angiotensin I increased signifcantly and returned to control levels when a normal salt diet was provided.  When another group of animals were provided a low salt diet and mercuhydrin, angiotensin I conversion decreased significantly, although returning the animals to a normal salt diet had no further influence upon conversion.  These experiments indicate that there is a direct relationship between total body sodium and the systemic conversion angiotensin 1.


Effect of diethylaminoethyl-dextran on the replication of a murine sarcoma (Moloney)-leukemia virus complex in mouse embryo cultures (38577).

Isolates of a sarcoma (Moloney)-leukemia virus complex prepared as cell-free extracts from mouse tumors showed no enhancement in infectivity by DEAE-D either in the sarcoma moiety measured by focus formation or in the leukemia moiety measured by XC cell assay.  The sarcoma moiety was not enhanced by DEAE-D in MEM and modified McCoy's 5a media or when varying amounts of FCS and glutamine were included in the media.  Progressive enhancement of viral infectivity by DEAE-D was found when the viral preparations were passaged serially in MEF cells.  DEAE-D ALSO ENHANCED TUMOR FORMATION IN VIVO BY TISSUE CULTURE PASSAGED VIRUS.


Physical, chemical and morphologic dimensions of human hepatitis A virus strain CR326 (38578).

CR326 human hepatitis A virus purified by isopycnic banding from infected marmoset sera was shown to consist of 27 nm spherical particles on electron microscopic examination.  The particles were identified as hepatitis A virus by tests for infectivity and by specific neutralization of infectivity with convalescent human hepatitis A serum.  Also, indentical 27 nm viruses in liver extracts gave specific reactions with hepatitis A antisera when tested by immune electron microscopy.  The bouyant density of the virus in CsCl was 1.34 and it was heat (60 degrees), ether and acid stable but was destroyed by heat (100 degrees), formalin (1:4000) and ultraviolet irradiation.  Electron microscopic studies of sections of infected marmoset liver showed intracytoplasmic virus particles, usually in vesicles.  Presumptive findings for RNA, together with the intracytoplasmic location of the virus, indicated the virus to be of RNA-type.  The attributes of the virus indicate it is closely related to the enterovirus family and not to hepatitis B virus.


Demonstration of feline leukemia virus antibody in cats by passive hemagglutination (38583).

Two FeLV fractions from Sephadex G-150 gel filtration were used to sensitize RBC for the PHA test. When the cells were coated with the fraction from the second peak consisting mostly of a mojor gs antigen of FeLV, no antibodies could be detected either in hyperimmunized cat sera or sera from leukemic cats, whereas antibodies were readily detectable in immunized rabbits. Using cells coated with the first peak eluants or Tween-ether disrupted FeLV, PHA antibodies were detected in cat sera. There existed, however, one significant exception that a cat with the diagnosis of mast cell leukemia showed antibody against the second peak fraction. Little or no antibodies could be detected in cat sera by CF or gel-diffusion. There was some correlation between hemagglutinating antibodies and conglutinating complement absorbing antibodies, but these antibodies did seem to differ from neutralizing antibody.


The renin-angiotensin system in rats made hypertensive by ligation of the kidney poles (38584).

The renin-angiotensin system was studied in experimental renal hypertension produced by ligation of the poles of the left kidney followed by contralateral nephrectomy.  Plasma renin concentration of renin substrate was lower and that of angiotensin I converting enzyme was higher in hypertensive animals. The juxtaglomerular index decreased in the medial zone of the kidney, while heavily granulated areas appeared in the poles.  Ligated kidneys of rats that remained normotensive showed juxtaglomerular indices intermediate between the control and the hypertensive rats.  Differences in renal renin content between the groups correspond to those for the juxtaglomerular index, but were smaller.  No differences between the experimental groups were observed in iso-renin content in the brain; however in all animals with ligated  kidney poles, hypertensive or normotensive, there was a tendency for iso-renin in the adrenals, left ventricular myocardium, and especially aorta to be lower than in controls.


The renin and iso-renin-angiotensin system in rats with experimental pitutary tumors (38585).

Renin, iso-renin, angiotensin I. angiotensin-converting enzyme, and angiotensinases were measured in plasma and in various extrarenal tissues of rats. Despite complete suppression of plasma renin in rats bearing pituitary tumors iso-renin and all other components of the renin-angiotensin system were found to be at or above control concentrations. The results strongly suggest that there is local synthesis of iso-renin in extrarenal tissues.


Effects of central dopaminergic blockade with primozide upon the EEG stages of sleep in man.

The effect of pimozide, a potent and specific blocker of central dopaminergic transmission, upon the sleep of man was studied in six healthy volunteers. Given at doses of 1 and 4 mg, which have clear central effects in humand. pimozide produced only minor changes in the EEG patterns of sleep. At these doses a slight and non-significant decrease in phase I sleep was observed, while phases W,II,III,IV and REM were not modified. No differences were noted between drug or post-treatment and control nights in total NREM sleep, total REM sleep, number of episodes of REM or total number of eye movements. An increase in REM sleep in the first REM period in the first drug night were the only statistically significant findings. If one accepts that central effects seen in man after pimozide given in conditions similar to those of this study are due to dopaminergic blockade, our results tend to suggest that dopamine is, at most, of rather minor importance in the physiology of sleep in man.


The effect of triazolam on the sleep of insomniacs.

The effects of three oral doses of bedtime triazolam (0.25 mg, 0.5 mg, and 1.0 mg) a new benzodiazepine, on the laboratory sleep of insomniacs were studied in a double blind design which used the following 14 consecutive night schedule: 1-4 placebo; 5-11 drug; 12-14 placebo. Effects on sleep were measured objectively by conventional EEG/EOG/EMG sleep recordings and subjectively by questionnaires administered each morning. Side or toxic effects were assessed by screening physicals and questionnaires administered each morning and each evening and by a comparison of the prestudy vs. end-study physical exams and clinical lab tests. At the 0.5 mg dose triazolam significantly reduced several objective and subjective measures of insomnia. It had lesser effects at the 0.25 mg dose and equal or greater effects at 1.0 mg dose. There were no remarkable side or toxic effects at any dose.


Quantitative determination of the abnormal lipoprotein of cholestasis, LP-X, in liver disease.

Quantitative determination of the abnormal plasma lipoprotein of cholestasis LP-X has been performed in 81 LP-X positive patients with different liver disorders. Great variations in the plasma concentration of LP-X were demonstrated both in the 45 patients with intrahepatic cholestasis (acute hepatitis, toxic hepatitis, primary biliary cirrhosis and cholangitis) and in the 36 patients with extrahepatic cholestasis (extrahepatic biliary obstruction by tumours and choledocholithiasis). The plasma concentratkons of LP-X in the patients with extrahepatic cholestasis (median 158 mg/100 ml) were significantly (psmaller than 0.001) higher than in the patients with intraphepatic cholestasis (median 25 mg/100 ml) was exceeded by 42% of the patients with extrahepatic biliary obstruction, and 33% of the patients with extrahepatic biliary obstruction, had LP-X LEVELS ABOVE 400 MG/100ML. The plasma concentration of LP-X was significantly correlated to the plasma activity of alkaline phosphatases and serum bilirubin, but seemed to be superior to these two parameters in the differentiation between intrahepatic and extrahepatic cholestasis. Plasma levels of LP-X above 400 mg/100 ml are highly indicative of extrahepatic biliary obstruction.


[Bone scintigraphy with 99m Tc pyrophosphate in neurology].

99MTc pyrophosphate has restored bone scintigraphy to the position of respectability it had lost with radiostrontium 85 and 87m, and it has rapidly become the method of choice for the detection of osteogenic hyperactivity. In cases of tumor the histologic disorder is frequently detected prior to the morphologic changes seen in X-ray pictures. The biophysical properties of 99mTc pyrophosphate provide high quality imaging of the skeleton. The scans or scintiphotos render possible a more sensitive and earlier diagnosis than radiography, though bone scanning does not of course supersede radiography. The two methods are complementary. In present day oncologic practice, however, bone scintigraphy with 99mTc pyrophosphate replaces to advantage the conventional radiographic "bone survey". In neurology the change is already appreciable and has proved its worth. In the light of results in 78 neurologic patients attention is drawn to the significant changes bone scintigraphy has brought about in diagnosis and therapeutic planning. The indications which have already emerged from this new method of nuclear neurology are tabulated, discussed and illustrated.


[Sclerosis of the intestinal lymphatic vessels].

Fibrosis of the intestinal lymphatic vessels, produced in one case by tuberculosis and, in the other, by appendicitis and peritonitis, caused blockage of the main lymphatic vessels causing, clinically, a protein-losing enteropathy similar to that noted in congenital lymphatic diseases of childhood. In the laboratory, there was noted a fall in serum protein, lipid and cholesterol. A fat absorption test was very abnormal showing a flat curve. During laparotomy, there was discovered on the small intestine, the same layout of lymph vessels, resembling a lace network, as that observed in congenital malformations. Intestinal lymphography showed considerable stasis of the opaque substance and absence of injection of the lymph vessels in the mesentery.


[Botulism. Therapeutic management. Apropos of 36 cases].

Botulism is relatively common in the Vienne department as we have observed 36 cases over a peroid of 7 years. Other French cases, published over the last 10 years, included 10 to 19 cases in the largest series. In a total of 10 series, there were 105 cases. Botulism is caused, in most cases, by eating ham prepared on the farm. It gives rise to the usual clinical picture including paralysis of accommodation, dysphagia, digestive symptoms with abdominal pain and constipation and finally, bladder paralysis. The mild forms are fairly common. We did not observe any severe cases. Our patients were treated in two homogeneous series of 17 cases each ; two patients were given no treatment. Patients treated by sero-anatoxin therapy according to classical methods, evolved towards a cure within 21 days on average. Patients treated by sulfguanidin were cured within 24 days. This study permitted us to confirm that sero-anatoxin therapy is not essential. The disadvantages are the number of injections, the possibility of sometimes fatal hyper-sensitive reactions which, in the opinion of many authors, justifies routine cover with cortisone. This attitude does not seem to us justified owing to the usually benign nature of the botulism. The treatment with sulfaguanidin is well tolerated and is not of any risk to the patient.


[Immunologic changes and infections in alcoholic cirrhosis].

The authors explored humoral and cellular immunity in 50 cases of alcoholic cirrhosis. The levels of the three serum immunoglobulins were greater than normal. IgM was significantly higher. The levels of antipolio antibodies (types I, II and III) were significantly higher in cirrhosis than in controls. The antistreptolysin and antistaphylolysin titers showed that the differences between levels found in cirrhosis and in controls were not significant. After taking the Sabin polio vaccine, variations in antipolio antibodies remained within normal limits, considering the high levels noted before taking the vaccine. Total serum complement was normal in 90% of the estimtions carried out. The results of the intradermal reactions were dissociated. Tuberculin skin tests were negative in 35% of cases of cirrhosis and in 14.3% of controls. In skin tests for candida and streptococcal antigen, differences between cirrhotics and controls were not significant. The absolute figure for blood lymphocytes per mm-3 was, in half the cases, less than normal.


[Epithelial cancer and monoclonal immunoglobulins].

The authors report 4 cases of carcinoma of the intestine, stomach, ovary and breast, association with monoclonal Ig. Clinical and laboratory analysis showed, in particular, that the immunoglobulin was present in small quantity and that there was no Bence-Jones protein in these 4 cases, that the bone marrow was little, or practically unmodified. The authors then discuss the nature, whether by chance or not, of this association the differential diagnosis with major monoclonal dysglobulinemia, e.g. myeloma or Waldenström's disease, and the significance in a cancer patient, of the discovery of Igmc. According to the authors, although in a few cases the tumour may induce development of a clone secreting Igmc, usually carcinoma and Igmc are independent from one another and are due to a common mechanism : reduction in cell immunity, according to Burnet.


Radioimmunoassay: review of basic principles.

The development of radioimmunoassay by the late Solomon A. Berson and Rosalyn S. Yalow during the late 1950s represents a milestone in the history of the application of radionuclide methodology to biology and to medical investigation and practice. The method offers a technique to assay materials otherwise unmeasureable or detectable only with difficulty. Radioimmunoassay is based upon the competition between labeled and unlabeled antigen for specific antibody sites, forming antigen-antibody complexes. This reaction is described by the expression see journal for formula. At equilibirum, the radioactive complex (bound) is separated from the radioactive antigen (free). The B/F ratio is dependent upon the amount of nonradioactive antigen. Antigen concentration in unknown samples is determined by comparing the B/F ratio to the B/F ratios obtained by incubating varying amounts of known nonradioactive antigen with the same amount of antibody as in the unknown sample under similar assay conditions. Sensitivity of the order of 10-12 moles/liter may be achieved through the preparation and use of a labeled antigen of high specific activity and the production and selection of antisera with appropriately high affinity constants. Specificity is dependent upon the ability of the antiserum to recognize subtle structural features of the antigen molecule. The ability to conveniently assay large numbers of samples with good precision has led to the application of this technique to quantitate substances (such as steroids) already measurable but by more cumbersome methods. Since the initial description of competitive binding radioassay techniques, there have been numerous contributions to its further development, refinement, and application. This article reviews the conception and development of this invaluable contribution to our understanding of health and disease.


Radioimmunoassay of insulin.

The powerful nuclear medicine tool of radioimmunoassay was very rapidly developed because of the favorable properties of insulin in such a procedure. The current method of insulin immunoassay in our laboratory is described, with emphasis on the details of the separation of bound and free using talc tablets; emphasis is also given to the correction for tracer damage. The clinical usefulness of the insulin radioimmunoassay is somewhat less than had been anticipated, although its use as a research tool has had immeasurable impact upon the study of metabolism. The presence of proinsulin in plasma is a factor that has to be considered during the radioimmunoassay. Radioimmunoassays for the connecting peptide of proinsulin have also been developed. A preliminary evaluation of two radioimmunoassay kits is described.


Radioimmunoassay of plasma renin activity.

Plasma renin activity is quantitated by measuring the rate of angiotensin generation during incubation of plasma renin with endogenous renin substrate. The angiotensin is quantitated by radioimmunoassay. Our studies indicate that the incubation step is best carried out in undiluted plasma at the pH optimum for renin (pH 5.7) in the presence of EDTA, neomycin, and DFP or PMSF. By using these conditions, incubation of low-renin samples can be prolonged for up to 18 hr, because angiotensinases and converting enzyme are completely inhibited. Accuracy is enhanced by prolongation of the incubation time, which results in more angiotensin and eliminates the need for blank subtraction. Incubation at pH 7.4 is disadvantageous because of lower rates of generation of angiotensin 1, because of the inability to maintain pH constant without addition of strong buffer, and because the incubation step cannot be prolonged beyond 3 hr. Dilution of plasma is undesirable because it results in a slower reaction rate due to dilution of both enzyme and substrate, and it is not possible to correct for the effect of substrate dilution. The radioimmunoassay of angiotensin I presents few unusual problems. The volume of plasma assayed should be 20 muI or less. If blank subtraction is used, antibodies should be screened to determine the extent to which they bind nonspecific substances in plasma, and then to ascertain whether the blank is entirely additive when angiotensin is added to it. Assay sensitivity is an important issue, since approximately 30 percent of patients with essential hypertension have subnormal plasma renin activity. In a study of three different commercial kits we found that many low-renin samples were undetectable and major fractions could not be discriminated with precision or consistency from normal renin samples. However, the incubation conditions can be easily modified, so that an 18-hr incubation can be utilized and then low-renin samples can be detected with the same degree of accuracy as those with normal plasma renin activity.


Physiologic consequences of hepatic lobectomy in man.

Replacement of glucose and albumin in ten patients after hepatic lobectomy shows that hypoglycemia and hypoalbuminemia, the two most common consequences of lobectomy in animals, can be prevented in man. Biosynthesis of protein, cholesterol, and prothrombin are reduced temporarily. In patients having emergency lobectomy, the serum bilirubin and glutamic oxaloacetic transaminase concentrations are statistically greater than in patients having elective lobectomy. Serum ammonia is not elevated and bromsulphalein excretion is normal after bilirubin returns to less than 1 mg/100 ml. Lactic dehydrogenase concentrations in serum are increased and fluctuate in the presence of sepsis or respiratory insufficiency: Mean creatine phosphokinase peaks at concentrations higher than those reported in acute myocardial infarction and returns to normal in three days. Compensatory hyperplasia of the residual  lobe occurred in all patients.


Somatic hybridization and oncogenesis; (Mechanism of formation of malignant tumors and metastases by the action of antilymphocytic serum).

The results of experiments carried out to test some of the consequences of the earlier general theory of oncogenesis, according to which the malignant tumor cell can arise as a result of somatic hybridization of cells of different organ- and tissue-specificity, are described. In the first series a tumor induced by cellophane film, was grafted into syngeneic and allogeneic mice, and antilymphocytic serum (ALS) was then injected. Metastases occurred only in allogeneic recipients receiving ALS. It was thus shown that the ability of cells of this particular tumor to metastasize is not a property inherent in its cells but is acquired by them as a result of interaction with the recipient organism. In the second series it was shown by two immunological methods that the cells of metastases arising under these conditions contain tissue compatibility antigens of donor and recipient origin, i. e., that they are somatic hybridsmin the third series skin from individuals of another strain was grafted on to mice and ALS was injected; hepatomas developed in 74% of these mice. The theory is used to explain several phenomena of carcinogenesis not explicable by other theories: the phenotypic nature of cell transformation, the causes and nature of the duration of the latent period of tumor development, the mechanism responsible for the ability of tumors to overcome the system of immunological defense, the mechanism of activation of endogeneous oncogenic viruses, etc. Finally an answer is given to the question: what is a tumor?


The effect of natural oestrogens on coagulation.

A study was undertaken to assess the effect of natural oestrogen on the coagulation profile of menopausal women. Forty-five postmenopausal women were selected at random and treated with conjugated oestrogen (PREMARIN) FOR ONE YEAR. Blood samples were tested before treatment, at the end of 3 and 9 months' therapy, and 1 month after treatment had been suspended. Parameters studied included: fibrinogen; platelet count and aggregation; single-stage prothrombin time; kaolin partial thromboplastin time; factors v and x assays and euglobulin lysis time. The only statistically significant alteration noted was a depressant effect on the platelet count. The significance of this change is commented upon. For practical purposes, it may be concluded that treatment of a group of menopausal women with conjugated oestraogen for one year had no adverse effect on their coagulation mechanism.


Incidence of paramalignant disorders in bronchogenic carcinoma.

The incidence of paramalignant disorders was studied in 280 consecutive patients with early lung cancer confirmed histologically. The commonest disorders were weight loss exceeding 6.4 kg (30.7 percent of the series), finger clubbing (29 percent), fever (21.1 percent), and endocrinopathies (12.1 percent). Paramalignant diseases are common even in early lung cancer and all patients should have investigations for their detection.


Disordered cholinergic neurotransmission and dysautoregulation after acute cerebral infarction.

The possible role of displaced neurotransmitter acetylcholine (ACHh) in dysautoregulation was examined after experimental regional cerebral infarction was produced by occluding the middle cerebral artery (MCA) in babons. Regional cerebral blood flow (rCBF) was measured after intracarotid injection of 133Xenon using the gamma camera. Autoregulation was tested with metaraminol or angiotensin infusion and the autoregulation index (A.I.) was calculated. Acetylcholinesterase (ACHhE) was measured in brain tissue of noninfarcted and infarcted hemispheres. Cerebral arteriovenous (A-V) differences for cholinesterase (ChE) were also measured. Regional dysautoregulation was found in infarcted gray matter and correlated with increased AChE levels in the same zones of cortex and basal ganglia. The time course of onset of dysautoregulation correlated with increased ChE uptake by the brain. Intravenous infusion of the cholinergic neurotransmitter blocker, scopolamine, restored autoregulation to the ischemic zones. Autoregulation appears to be a myogenic reflex, influenced by neurogenic and metabolic mechanisms.


Immunofluorescence detection of casein in human mammary dysplastic and neoplastic tissues.

To detect and localize casein in human mammary dysplastic and neoplastic tissues, an indirect immunofluorescence method has been devised. Anti-casein antibodies have been obtained from rabbits immunized with casein isolated from human milk. Cryostat sections post-fixed in alcohol and paraffin sections from routinely formalin-fixed tissues proved suitable. The immuno-fluorescence method revealed the presence of casein in epithelial cells in mammary dysplasia and in some, but not all, cases of carcinoma of the breast. Well differentiated (Grade I) carcinomas were shown to contain casein, mainly localized at the inner border of the epithelial cells; the milk protein was present also in cells metastatic in lymph nodes. In Paget's disease of the nipple, casein could be detected in neoplastic cells in the ducts and also in cells identifiable with the typical intra-epidermal Paget cells. This finding is evidence of a functional differentiation of Paget cells along lines proper to the mammary epithelium.


Malignant mixed osteogenic tumours of the breast. An ultrastructural study of two cases.

Two cases of malignant mesodermic tumours of the human mammary gland with osteogenic and chondrogenic structures were analysed by optical and electronic microscopical means. One of them was associated with an adenocarcinoma of the gland. The histological pattern was similar to that of those cases previously examined under the optical microscope in the mammary gland and in extraskeletal osteogenic sarcomas of soft tissues. When investigated under the electron microscope, the chondroblasts possessed a highly developed RER in active synthesis with an amorphous material which contributed to the building up of the ground substance matrix of the tumorous cartilage. Osteoid fields with scattered osteoblasts appear throughout the tumorous stroma and were associated with calcium deposits. They were continuous with fibroblasts and mesenchymal undifferentiated cells of a very immature character. Giant cells of osteoclastic type were included within the mononucleated mesenchymal cells mimicking bone osteoclastoma. The presence of all these cell types suggests the existence of a common malignant origin, the stem cell being differentiated into epithelial carcinomatous and mesenchymal sarcomatous chondral and osteogenic tissues.


[Intraepithelial filament bundles of the adult human kidney. Studies by light and electron microscopy and by diffraction analysis (author's transl)].

In studies of semi-thin sections of adult human kidney it is found that the structures formerly known as "Basalreifen" can be definitely related to basal cytoplasmic portions of the renal epithelium. Under the electron microscope they appear as bundles of filaments, usually having a width of 50-85 A. They are particularly well developed in the parietal layer of Bowman's capsule and are also present in the proximal and distal tubule where they are more abundant than in other parts of the nephron. The filament bundles proceed toward hemidesmosome-like structures at the basal border of the epithelia and sometimes contain dense bodies in the parietal capsular epithelium of the glomerulus; these bundles are probably responsible for local intra- and/or supravital protrusions of the cytoplasm. Findings in some diseased kidneys are also presented. Diffraction analysis of intrarenal vascular muscle cells and intraepithelial filament bundles gave comparable photographic records of the image structure spectrograms; it supports the assumption derived from the electron microscopic results that the intraepithelial filament bundles represent a contractile complex. However, the conclusiveness of image structure analysis in comparing the two biological objects is limited, as outlined in the discussion. With regard to renal function the significance of the intraepithelial filament bundles is still unclear. Periglomerular interstitial cells, which normally contain only narrow tracts of filaments in the periphery, had many of the characteristics of smooth muscle cells in a case of pyelonephritic renal cirrhosis. They are compared with myofibroblasts of a different origin.


Red cell esterase D in studies of paternity cases in the United Kingdom.

The recently discovered genetic polymorphism of human red cell esterase D has been applied to 156 cases of disputed paternity, along with other well-established systems. The results indicate that esterase D is a useful additional genetic marker for use in paternity testing.


Unsuccessful trial of gene replacement in arginase deficiency.

The Shope virus following cutaneous inoculation produces skin papillomas in rabbits and induces a virus-coded arginase in the squameous epithelium of these papillomas. The only discernible effect of the virus inoculated intravenously in rabbits and other animals was a decrease of the serum arginine concentration. Intravenous injection in 3 hyperargininemic patients, however, did not influence the underlying metabolic disease.


[Serological variants of type 1 parainfluenza viruses].

A comparative study of the antigenic properties of parainfluenza type 1 viruses revealed the existence of 2 serological variants among them. The expedience of using the reference and recent years of isolates of parainfluenza type 1 viruses for serodiagnosis was demonstrated. Highly immunogenic strains isolated in 1971 were selected for properation of hyperimmune sera. It is necessary for identification of new isolates of parainfluenza type 1 viruses to use antisera for strains isolated in recent years alongside with reference immune sera.


[Isolation of intact liver parenchymal cells by a modified enzymatic method].

An enzymatic method is described for isolating intact parenchymal cells from rat livers. 3--4 g cells (wet weight) could be isolated from livers of rats weighing 180--230 g. After an in vitro preperfusion of 15 minutes with a Ca-free buffer, collagenase (200 mg/1) and calcium chloride (5.2 mmol/1) were added. Perfusion was continued for another 15 minutes at 37 degrees C. Micromorphological integrity of cell membranes was demonstrated by scanning electron microscopy. With regard to rates of gluconeogenesis and protein synthesis, parenchymal cells isolated according to our method were found to be superior to liver slices and cells isolated by other methods. Ratios of ATP/ADP (5.69) and of lactate/pyruvate (8.64) as parameters of the energetic situation and the redox state resp. were found within the physiological range. Integrity of cell surface receptors was proved by their sensitivity to epinephrine, glucagon and insulin. Glucagon (0.3 mumol/1) and epinephrine (1 mumol/1) and reduced glycogen deposition in hepatocytes of fasted rats by 84.9 % and 95.9 % resp. Both hormones stimulated glycogenolysis in parenchymal cells of fed rats to a similar extent. Urea synthesis was stimulated 29.5 % by glucagon (1 mumol/1), and inhibited 28.0 % by insulin (10 nmol/1). The stimulatory effect of glucagon (1 mumol/1) was abolished by insulin (10 nmol/1).


[An enzymatic method for the isolation of tubules and cells from human kidney cortex].

The isolation of tubules and cells from human kidney cortex was realized by an enzymatic method. Tubules and cells were released from slices of kidney cortex by collagenase. The yield amounted to 80 % of the wet weight of incubated cortex slices. Thus numerous experiments with isolated tubules from one organ could be performed. Glucose production from different substrates was measured in order to test the biochemical integrity of the isolated cells. The highest rates of glucose formation were obtained with fructose as precursor. Glucose production was higher from lactate than from pyruvate. With proline and glutamine as substrates only small amounts of glucose were produced. Glucose formation from 10 mmol/1 pyruvate was linear with time up to 80 minutes. Ado-3':5'-P stimulated glucose formation at 10 mumolar concentration and inhibited gluconeogenesis at 1 mmolar, 0.1 mmolar and 1 mumolar concentrations.


[Infectious allergy. XIV. Histochemical changes in redox enzymes in cells of the small intestine smooth muscle during sensitization with diphtheria exotoxin].

Administration to guinea pigs of 1/Dlm of diphtheria exotoxin twice at an interval of 1 and 24 hours, or once in a dose of 25/100 Dlm caused changes in the activity of the redox enzymes. The character of their changes in the animals to which the whole dose of the exotoxin was administered once or repeatedly was directly reverse: in the first case the activity of the enzymes was uniformly increased, and in the second--ununiformly depressed; besides, there was a marked disturbance of coordination between the enzymes. An increased sensitivity of the infectious allergy type following injection of low doses of diphtheria exotoxin was not accompanied by immunity formation. Specific antibodies against diphtheria exotoxin formed only in the organism of animals to which 1/100 Dlm of diphtheria toxin was injected once a day for 15 days, but in this case as well no immunity against diphtheria exotoxin was observed.


[Ultrastructure of L-forms. II. L-forms of Listeria monocytogenes].

A study was made of ultrathin sections of the stable l-forms of listeria obtained under the action of penicillin in meat-peptone-liver broth. A marked cellular polymorphism was found in the L-form culture: within the same colony cells differed in size, shape and fine structure. It is supposed that polymorphism could be partially explained by a different plasticity and premeability of cytoplasmic membrane in different types of cells of the same L-colony. The three-layer structure of the membrane does not always display the same distinctness in various L-colony cells and also in different areas of the cell surface. Structureless material of low electron density, possibly a defective murein or its precursor, was revealed on the membrane surface. Electrondense inclusion bodies, mesosomes of ring-shaped or more complicated structure and two-contour vesicles were found in the cytoplasm. The cells multiplied by budding, by binary and anomalies division participation of mesosomes in this process was not proved by the L-forms.


[Characteristics of the low molecular antigens of pertussis bacteria].

The authors elaborated a method of obtaining pertussis soluble antigenic complex by dialysis through the cellophane membrane against the physiological saline at a temperature of 4 degrees C. An antigen which was active in the passive hemagglutination and neutralization of antibodies tests was revealed in the dialyzate. The amount of this antigen in the dialyzate increased gradually up to the 7th day and then became stabilized. The serological activity of the antigen after evaportation increased 4-16 times. The results of the antibody neutralization test pointed to the presence in the dialysate of substances common to those contained in the 1a and 1Da fractions isolated from the pertussis bacteria with the aid of ammounium sulfate.


[Biological properties of plasma-negative staphylococci isolated from patients in surgical departments].

A study was made of 111 strains of plasma-negative spathylococci isolated from the blood, pleural fluid, urine, and exudate of the abdominal cavity of 30 patients. The studies were carried out by 18 criteria. A variety of biological properties and signs characteristic of pathogenic staphylococci (hemolytic activity, anaerobic splitting of mannite, the presence of phosphatase, lysozyme, protease, alpha-toxin, fibrinolysin) were noted. A high resistance to tetracycline and penicillin was found in the strains isolated from the blood and the pleural cavity.


[Use of a synthetic medium for cultivating pathogenic anaerobes].

It was shown that a synthetic medium suggested by the authors earlier was useful for the growth and toxin formation of Cl. tetani, Cl. botulinum and Cl. perfringens, types B and E. A study of the character of growth and toxinogensis, microscopic examination of morphology of culture cells and results of passages showed the suggested synthetic medium to be of value; a possibility of its application for studying the nutrient requirements and the role of individual components of the nutrient media in the process of growth and toxinogenesis was also demonstrated.


[Comparative study of nutrient media for isolation of the agents of pertussis and parapertussis].

Taking into consideration literature data pointing to a possibility of a marked improvement of the quality of the medium for the isolation of the causative agents of pertussis and parapertussis, an experimental study was made of the sensitivity of the nutrient media produced by the Dagestan Institute of Nutrient Media, a medium made from dry and also of a number of media modified by the authors prepared on its basis. Pure freshly-isolated and museum B. pertussis cultures were used in the observations. Analysis of the results of a comparative study of the indices (detectability of the strains, the appearance of colonies and periods of their formation) allomed a recipee of the most sensitive medium to be elaborated. At the same time there were found considerable differences in the indices of the appearance of B. pertussis colonies and detectability of the museum cultures used for testing the industrial batches and of the freshly-isolated strains.


[The response of blood neutrophils (the PPN test) to pertussis allergen in children with pertussis and children immunized with ADPT vaccine].

The authors present materials on the study of the reaction of blood neutrophils (PPN test) to pertussis allergen in children suffering from pertussis and immunized with ADPT vaccine. Results obtained in examining 111 children showed that the PPN test was specific and could be used for assessment of allergic manifestations in children suffering from pertussis or immunized with ADPT vaccine. Taking into consideration the harmlessness and expressiveness of the PPN test it can be recommended for studying in dynamics in any age groups.


New kind of cytoplasmic inclusions of plasma cells in acid maltase deficiency.

Extensive 'vacuolization' could be demonstrated in nearly all plasma cells and in some lymphocytes of an adult with glycogenosis type II (Pompe's disease). The biochemically defined diagnosis acid maltase deficiency (AMD) could be ascertained by examination of the maltase activity of the patient's leukocytes. Electron microscopical, microspectrographic, and cytochemical investigations revealed electron dense inclusions, which show an UV absorption at 276 nm and a positive reaction after PAS staining of plastic embedded material. Since no other abnormalities of the plasma cells could be found, our results are presumably indicative for a connection of AMD and a glycoprotein storage in the plasma cells of the patient.


Fenestrae in golgi and endoplasmic reticulum cisternae of human brain tumours.

Fenestrae were found in freeze-fractured cisternae of the Golgi apparatus and endoplasmic reticulum of glioblastoma, oligodendroglioma, ependymoma, medulloblastoma, medulloepithelioma, meningioma, cerebellar sarcoma, hemangioblastoma, and chromophobe adenoma. They were about 200--400 A in diameter and often diffusely distributed or concentrated in groups in Golgi cisternae, while they were around 300--600 A in size and scattered in distribution in cisternae of endoplasmic reticulum. They appeared as conical protrusions or circular broken-off necks of face A and as circular holes on face B in tangential fractures, and as several constrictions of cisternae in cross fractures.


Morphological observations of dysplastic gliomas heterotransplanted to experimental animals.

Observations concerning transplantation of 5 cases of malignant dysplastic gliomas to the anterior chamber of the guinea-pig eye have been presented. This type of gliomas, composed mainly of cytoplasm-abundant cells, show weak transplantability, slow rate of growth, and particular tendency to degeneration and necrosis. At the same time, in subsequent passages, an intensive proliferation of small-cell component occurs, comparing with cytoplasm-abundant cells.


Atypical astrocytes and Rosenthal fibers in a case of amyotrophic lateral sclerosis associated with a cerebral glioblastoma multiforme.

A case of amyotrophic lateral sclerosis (ALS) and cerebral glioblastoma multiforme is presented in which bizarre astrocytes were found in the degenerating lateral corticospinal tracts, along with Rosenthal fibers, which were present in the corticospinal tracts of the lower medulla and spinal cord, and in anterior horns. These bizarre astrocytes did not result from direct infiltration of tumor from the cerebrum. "Malignant transformation" and/or an exceptionally intense glial response to the corticospinal tract degeneration are discussed as possibilities for the development of these bizarre astrocytes. Rosenthal fiber formation is described, to our knowledge, for the first time in ALS.


Studies on the presence of antibodies to EB virus and other herpesviruses in normal children and in infectious mononucleosis.

Children free from infectious disease have been examined by indirect immunofluorescence for the presence of antibodies to the intracellular capsid antigen of Epstein-Barr virus (EBV). In the first year of life 46%, between 2 and 6 years of age 66%, and between 7 and 14 years 91%, of the children proved positive. The corresponding percentages for the presence of antibodies to cytomegalovirus (CMV) and herpes simplex virus were about 50%, irrespective of the children's age. Serum samples from 69 patients suffering from infectious mononucleosis (IM) were tested for anti-EBV antibodies. Of the 29 Paul-Bunnell-positive patients 22 had antibodies, 11 of them in high titres (greater than 1 : 80). Of the 40 Paul-Bunnell-negative cases only 21 had antibodies, 8 in high titres. Of the Paul-Bunnell-negative cases, 73% were found to have anti-CMV antibodies, 32% in high titre. The respective percentages for the Paul-Bunnell-positive cases were 42% and 10%.


Factors influencing the stability of acid-precipitated polyvalent Bordetella pertussis bulk suspensions.

According to the statistical analysis of the potency values of acid precipitated polyvalent bulk suspensions of Bordetella pertussis, the potency of suspensions decreased below 8 IU/30 IOU during storage for 5 to 9.8 years. The average annual decrease over a 13-year period was 2.03 IU/30 IOU. The periodicity in the decrease of potency values was assumed to be connected with changes in the quality of the casein hydrolysate ingredient of the medium. Periodic functions employed to approach potency values with limits of 1 SD falling within the range of 64--156% indicated that the potency decreased below 8 IU/30 IOU after 9 years.


Managing ischemic skin ulcers.

Etiologic elements in ischemic ulcers include physical factors (pressure, shearing forces, friction and heat), nutrition and infection, as well as circulatory, metabolic and neurologic problems. Prevention is the primary objective of the management program. For patients in nursing homes who have already developed ulcers, low-intensity direct electric current (LIDC) has proved to be a useful adjunct to conventional management. The infection was more rapidly controlled and the wound size more quickly reduced. LIDC is readily accepted by the patient.


An unusual membrane-bound body in rat periosteal cells.

Cytoplasmic inclusions have been found in both normal and stimulated rat periosteal cells. These inclusions are bounded by a pentilaminar membrane similar to a tight junction and contain a core of granular material. The origin and function of this inclusion are unknown.


Passage of red blood cells through the sinusoidal wall of the spleen.

The migration of blood cells across the wall of splenic sinusoids has been studied by means of serial sections in rats inoculated with murine erythroblastosis virus (MEV). The sinusoidal walls of the normal spleen have no permanent openings. In the normal spleen, mature red blood cells and blood cells in the latest stages of maturation can be seen in diapedesis. The predominant form of transmural migration is intercellular. In MEV-inoculated animals, in which there is a vigorous erythroblastic proliferation, large numbers of erythroblasts are in transmural passage. Serial sections show that these cells migrate through the cytoplasm of the endothelial lining cells. On the basis of these observations, it is suggested that the transcellular mode of migration may be related to the delivery of newly formed red blood cells into the circulation while the intercellular mode may be part of the mechanism by which the spleen controls the removal of red cells from the circulation.


Plasma alpha-lipoprotein pattern in acute viral hepatitis.

In the present study, the plasma levels of lipoproteins, triglycerides, and cholesterol were followed serially in a group of 57 patients with acute viral hepatitis. Mean plasma triglyceride levels were found elevated at the onset of the disease and gradually returned to normal, while mean plasma cholesterol values, low initially, gradually increased, alpha-lipoprotein was absent at the early stage of hepatitis in 41 out of 46 patients with mild or moderate course and reappeared gradually during the course of the disease. In 11 cases of viral hepatitis with impending or overt coma, alpha-lipoprotein was absent for the whole duration of the acute stage and never reappeared in those who eventually died, while it eventually returned to normal in those who survived. The reappearance of alpha-lipoprotein in acute viral hepatitis appears to be a sensitive index of improvement and a significant prognostic sign.


Acute respiratory disease of university students with special reference to the etiologic role of Herpesvirus hominis.

Infections with Herpesvirus hominis type 1 were associated with 11.5% of acute respiratory illnesses of university students who were admitted to the student infirmary over a 6-year period. Over three-quarters of these infections were detected in students with pharyngitis or tonsillitis; 42% had ulcerated lesions on tonsils or posterior pharynx but only 11% had lesions in the anterior portion of the mouth or lips. Almost all of the H. hominis infections were accompanied by significant rises in neutralizing antibodies and few students had detectable antibodies in the initial serum collected during the acute phase of illness. Special studies revealed herpes-specific IgM antibodies in the early convalescent sera of some of these patients. The data demonstrate that 80% of the infections detected were primary infections with H. hominis. Only 30% of university students possessed neutralizing antibodies to H. hominis and about 10% of those without antibodies acquired antibodies each year. These data suggest that the majority of persons from middle income families reach young adulthood without acquiring infections of H. hominis and the spread of the virus requires close and intimate contact.


The Seattle virus watch. V. Epidemiologic observations of rhinovirus infections, 1965-1969, in families with young children.

Rhinovirus (RV) infections in Seattle Virus Watch (VW) families (1965-1969) were monitored by screening respiratory specimens in WI-38 cell cultures and by homotypic neutralization tests on sera related to family episodes revealed by RV isolation. Temporally related illness in members not proven infected was also taken to indicate infection. RV isolates (including those from the New York VW, 1961-1965) were typed within the official 90-serotype frame. Typed isolates from New York (165 with 39 serotypes) and Seattle (456 with 59 serotypes) were compared with the Tecumseh Study to test the hypothesis that some serotypes are "common," persisting because of greater infectivity. Of 32 serotypes qualifying as "common" in at least 1 study, 4 were "common" in all 3 studies and 8 in 2 studies. The 23 "common" Seattle serotypes differed from the remaining 36 serotypes in being more infective and in their more frequent association with prolonged shedding. The New York and Seattle isolates together revealed an increase over time in the proportion not typable or of of higher numbered types, consistent with progressive shift in RV antigenic character. WI-38 isolates indicated spring peaks of RV all 4 years but a fall peak only in 1967. An even larger fall peak was seen when all specimens from September-November 1968 were re-examined in fetal tonsil diploid cells. Thus, both spring and fall peaks appear to describe RV seasonality. RV infections explained 16% of all reported respiratory illness (20% of upper respiratory), but RV-associated illness in young children, especially under 2 years, was more severe and almost twice as frequent as in adults. The age of introducers and the direct relation of family size to frequency of episodes indicate that community spread depends largely on preschool children, including infants. Within families, the secondary attack rate (SAR) was highest following paternal introduction and, for all introducers, the SAR varied inversely with age (mother excepted). RV shedding was observed most often (85% of specimens) from the day before to 6 days after illness onset but prolonged shedding was common (to 21 days in 20% and 28 days in 1.4% of infections). RV infectivity, reflected by SAR among nonimmunes, was highest for infants (78%) and, for all ages, was greater with ill than with well introducers (71% versus 27%). Immunogenicity of RV was poor (Seroresponse: 48% of shedders, 32% of nonshedding contacts) but varied greatly with serotype. Illness frequencies among non-immunes were 59% for all proven infections and 35% when infection was not shown..


Seroimmunity to poliomyelitis in an American community.

In an effort to determine the immune status of individuals in a natural community, neutralizing antibody titers for the three types of poliovirus were examined in 301 randomly sampled sera collected in 1966 and 308 specimens from 1971. The specimens were part of a larger collection obtained from residents of Tecumseh, Michigan. Approximately 22 sera from each five-year age group between 5-70 years were tested. A somewhat smaller number from younger and older individuals were also studied. Geometric mean titers for the two years of collection were in general quite low and never exceeded 1:42 which was found for type II virus. All GM titers decreased markedly as the age of the subjects increased. The percentage of individuals lacking antibody at the 1:4 level to at least one type of poliovirus ranged between 20-30% until age 49 and then steadily increased to over 60% in the 60-69 and over 70-year groups. A retrospective history of vaccination was obtained from 264 of the 308 persons in the 1971 collection. As expected these showed rather poor correlation with the presence or absence of antibody and were considered to be unsatisfactory as an index of immunity.


Familial occurrence of Wilms' tumor: nephroblastoma in one of monozygous twins and in another sibling.

We report the occurrence of pathologically documented Wilm's tumor in a 24-month-old male twin and just 9 months later in his 12-month-old male sibling. We considered the twins to be monozygotic because of their phenotypic similarities, the probability computed from analysis of blood groups, and the comparison of their dermatoglphics. There were no other persons in the kindred with either tumor or associated malformations, and the parents were not consanguineous. Because of the frequency of Wilm's tumor, the few instances of demonstrated occurrence in siblings seem insufficient to postulate monogenic determination. Concordance in monozygotic twins has simply not been proven. The monozygotic unaffected twin of our first patient has remained without evidence of tumor to 5 years, and, as long as he remains so, he appears to represent an exception to the hypothesis of the mutagenic origin of this childhood tumor.


Hypogonadism in Prader-Willi syndrome.

Sexual development was evaluated in 9 female and 2 male subjects with Prader-Willi syndrome. The process of sexual development and degree of genital development attained were found to be variable but abnormal in all subjects. Hypothalamic-pituitary-gonadal functions were evaluated by measurement of serum Luteinizing Hormone and plasma testosterone responses to stimulation by clomiphene citrate and plasma testosterone responses to stimulation by human chorionic gonadotrophin. The degree of vaginal estrogenization was variable. The testicular biopsies showed abnormalities mainly in the germinal epithelium. In agreement with previous studies, it was concluded that the abnormalities of sexual development in this syndrome are mainly due to a defect in the hypothalamic pituitary axis. Adrenal function was not found to be grossly abnormal. The 17 ketosteroid excretion values were low, probably explaining the rather sparse pubic and axillary hair observed in these patients. The urinary 17-hydroxycorticosteroid creatinine ratios were found to be elevated, probably due to decreased creatinine excretion, reflecting the muscular abnormalities of these subjects.


Partial unilateral cryptophthalmos with syndactyly, brachycephaly, and renal anomalies.

The cryptophthalmos syndrome consists of cryptophthalmos, dyscephaly, syndactyly, and genitourinary abnormalities. A 2-year-old girl had unilateral partial cryptophthalmos associated with brachycephaly, unilateral syndactyly, and unilateral poor renal visualization.


A cytoplasmic body in human fetal endothelium.

We describe a crystalloid-containing inclusion in the cytoplasm of human fetal cells believed to be endothelium. We do not know whether it is peculiar to the fetal period, nor do we know its function.


An experimental mouse testicular teratoma as a model for neuroepithelial neoplasia and differentiation. I. Light microscopic and tissue and organ culture observations.

The various stages of divergent neuroepithelial differentiation were studied in the solid transplants of a transplantable mouse testicular teratoma (OTT-6050) maintained in both ascitic and solid forms. They included: a) areas of undifferentiated medullary epithelium corresponding to the rare human medulloepithelioma; b) areas of neuroblastic differentiation corresponding to neuroblastoma, with more mature neuronal differentiation corresponding to ganglioneuroma or, when mixed with glial elements, to ganglioglioma; and c) more mature neuroglial areas resembling astrocytoma, oligodendroglioma or ependymoma, as well as more primitive areas corresponding to ependymoblastoma. In tissue culture using collagen-coated coverslips, astrocytic differentiation was found in the outgrowth zone after 15 days, confirmed by immunofluorescence with antibodies to an astroglia-specific protein. In organ culture systems, glial components, including ependymal structures, were preserved in tumor explants, and astrocytic differentiation, as expressed by glial fiber formation, was increased after 4 to 6 weeks in vitro. No neuronal differentiation was demonstrable, however. The neuroepithelial component of this experimental teratoma may provide a model for the study of neoplastic neuroepithelial differentiation.


Light and electron microscopic studies on human temporal arteries with special reference to alterations related to senescence, atherosclerosis and giant cell arteritis.

Temporal artery biopsy specimens from 26 patients of various ages with and without giant cell arteritis afforded an opportunity to examine several ultrastructural features of these human muscular arteries, including senescent and atherosclerotic alterations and the fine structural pathology of temporal arteritis. The unusual pathologic features of temporal arteritis were found superimposed on the progressive accumulation of smooth muscle cells, collagen and occasional discrete intimal atherosclerotic plaques in the intima of aging arteries. Two features of giant cell arteritis were conspicuous: first, the accumulation of large numbers of histiocytes and epitheloid and giant cells at the intimal-medial junction and second, fragmentation, degeneration and dissolution of the internal elastic lamina. The close proximity of the granulomatous reaction to the degenerating lamina suggests that these two aspects of the pathologic picture are in some way related, and possible immunologic mechanisms of this relationship are discussed on the basis of the ultrastructural findings.


A progressive glomerulosclerosis occurring in partial five-sixths nephrectomized rats.

A chronic renal disease was induced by 5/6 nephrectomy in young adult Wistar albino male rats. Groups of 3 rats were killed at varying time intervals from 10 to 50 weeks after nephrectomy, and the glomerular structure was studied by light and electron microscopy. By light microscopy, increase in glomerular size and hypertrophy of the visceral glomerular epithelial cells were evident as early as the tenth week. Significant degree of glomerular hyalinization started on the 25th week and gradually became more extensive in involvement towards the 50th week. By electron miscroscopy, the hypertrophied glomerular epithelia showed many osmiophilic bodies and vacuoles in their cytoplasm and fusion of foot processes. Beginning on the 30th week, increase of the mesangial matrix was quite evident, and endothelial and epithelial cells disappeared from the areas where the increase was prominent. The increased matrix gradually encroached and occluded capillary lumina and Bowman's space, leading to a formation of obsolescent glomeruli. Whether or not these glomerular changes are due to the glomerular hyperfiltration which has been known to occur after 5/6 nephrectomy remains to be elucidated.


Factors affecting long-term survival of patients with bronchogenic carcinoma.

From January 1963 to December 1968, 148 patients underwent thoracotomy for bronchogenic carcinoma. In 123 patients either lobectomy or pneumonectomy was performed (resectability rate of 84 per cent). The over-all operative mortality was 3.4 per cent. Forty of the patients undergoing resection (34 per cent) are alive and free of cancer five years after surgery. Tumor size, nodal involvement, cell type, location, symptoms, and extent of surgery were studied in relation to the long-term results. Five year survival was directly related to the size of the tumor and the extent of nodal involvement. No patients with mediastinal nodal involvement or with lesions larger than 7 cm in diameter were among the long-term survivors.


Postanal changes in the localization of glucose 6-phosphatase activity within the liver lobule of the mouse.

The change in the localization of glucose 6-phosphatase activity within the liver lobule of the mouse was studied during development. From one day before birth to three days after birth, a uniform localization of the activity was observed throughout the lobule. This uniform localization of the acitvity gradually changed to that of the adult type, characterized by a relatively higher activity in periportal areas, between three to ten days of age. This indicates that the postnatal growth of the liver is accompanied by a change in the pattern of the localization of this enzyme activity within the liver lobule. Further, the biochemical results showed that the elevated activity after birth was reduced to the level seen in adult at ten days of age. Thus, the reduction in the level of the enzyme activity during ten days after birth corresponds to a redistribution of the enzyme activity within the liver lobule.


Immunity to fowlpox.

Protection against fowlpox develops following vaccination or natural infection. The immune response can be measured for variable times either by challenging immunity of vaccinated birds for protection or by various serologic tests, e.g., viral neutralization, agar gel precipitation, complement fixation, passive hemagglutination, fluorescent antibody, and immunoperoxidase methods. Variation in the antibody response measured by these tests may be due to concentration of antibody present and sensitivity of the test employed. Available information suggests that both humoral and cellular immunity are involved in protection. There is a possibility of persistence of virus in latent infections which also might be responsible for protection.


Avian viral arthritis: antigenic types and immune response.

Serums from broiler breeders of an integrated broiler operator were examined for precipitin antibodies using antigens prepared from preliminary reovirus serotypes represented by WVU-2937, Reo-25, Reo-59, and Fahey-Crawley virus. The precipitin antibodies were present in most of the flocks. Virus neutralization tests indicated that the reovirus responsible for viral arthritis represented by WVU-2937 was not present in the breeder flocks, but antibodies to Reo-25, Reo-59, and the Fahey-Crawley viruses were present.


Immunity to transmissible (coronaviral) enteritis of turkeys (bluecomb).

Turkey flocks recovering from natural and laboratory induced coronaviral enteritis developed lifelong immunity. Virus neutralization tests indicated that neutralization capacity of serums from recovered turkeys was low. One-way cross challenge test using turkeys inoculated with various bluecomb isolates revealed close antigenic relationship or antigenic identity among bluecomb isolates. The controlled application of intestinal preparations containing coronavirus as a vaccination procedure had value in developing active lasting immunity but it has limitations in areas that are attempting to eliminate the disease by a depopulation program. Killed vaccines injected parenterally did not produce a detectable immunity.


Ulcerative enteritis--clostridial antigens.

The causative agent of ulcerative enteritis (UE) is a clostridium and it is believed that it belongs to a new, hitherto undescribed species. It has been named, tentatively, Clostridium colinum. Transmission experiments carried out with artificial cultures of this clostridium have clarified some aspects of the pathogenesis of UE and have brought into focus other aspects of its pathogenesis, epizootiology, and immunology that need to be investigated. Support for research is needed: 1) To investigate the distribution in nature of C colinum. For this purpose the development of more suitable selective and differential culture mediums should be encouraged. 2) To further study the pathogenesis of UE through application of the fluorescent labeled antibody procedure. 3) To search for toxins produced by this clostridium, possibly through use of ligated intestinal loops in chickens. 4) To develop a quick and simple means for the differential diagnosis of UE. Use of a fluorescent antibody test has been suggested as an area of research to fill this need.


Experimentally induced respiratory syncytial viral infection in calves.

Calves exposed by intranasal (IN) or intratracheal (IT) or both routes of inoculation to bovine respiratory syncytial (BRS) virus had a transient increase in body temperature for 2 days. Histopathologic changes indicative of pneumonia, along with multinucleated giant cells, were observed in alveolar lumen. The virus was recovered from infected calves, and their serologic response was poor. In a limited serologic survey with 100 serums from cattle in Maryland, low levels of serum-neutralizing (SN) antibody to the virus were prevalent in 38% of the serums.


Immunity and the level of neutralization antibodies in foals and mares vaccinated with a modified live-virus rhinopneumonitis vaccine.

Foals (farms 1 and 2) and mares (farms 1 and 3) vaccinated with a commercially available rhinopneumonitis vaccine were tested for the level of antibodies and for protection against the natural infection. The serum-neutralization (SN) antibody titers against equine herpesvirus type 1 were 1:8 or less in the majority of foals and 1:16 or higher in about 10%. The level of the nasal secretory antibody in the foals (farm 2) was less than 1:4. The vaccinated foals did not show any apparent respiratory tract infection for about 6 months and then an epizootic of respiratory tract disease appeared on farm 1 involving 3 foals. From each of these 3 foals herpesvirus was isolated. On farm 1, about 50% of the vaccinated mares aborted, and on farm 3, there were no abortions. Isolations of herpesvirus were made from 5 of the 6 aborted fetuses (farm 1). The SN antibody titers of the pregnant mares in the 2 farms at the time of foaling were high.


Colon and rectal cancer in the young adult.

801 patients with colon and rectal cancer were studied to assess the behavior of this cancer in the patient under 40 years of age as contrasted to the more commonly seen older patient. The younger patient had a greater frequency of advanced signs, later stages of cancer and mucoid carcinoma. However, when compared by clinical staging, the younger patient did as well or better than his older counterpart. Clinical staging was the most important prognostic factor irrespective of age. No inherent difference was found in the virulence of the cancer in the young, as the five-year survival in the younger patient (31 percent) was essentially the same as in the older patient (32 percent).


Pathophysiology of acupuncture: nervous system transmission.

As far as we know, this is the first scientific report which shows distinctly that acupuncture is transmitted directly by the nervous system and the performance of analgesic acupuncture requires an intact nervous system. The more distal the acupuncture treatment is given upon the nerve, the greater the degree of analgesia is afforded. The slowing from a normal awake beta to a normal awake alpha electroencephalographic rhythm may have some effect on the patient's pain perception but further investigation is required.


Surgical intervention in intestinal amebiasis.

Operative intervention in patients with unrecognized and untreated amebomas or acute fulminating amebic colitis may result in severe complications. Nevertheless, certain clinical presentations of intestinal amebiasis require surgical procedures varying from drainage of an abscess to a subtotal colectomy. If, in patients with acute fulminating amebic colitis, the signs or symptoms of intraperitoneal or impending perforation develop, unremitting diarrhea associated with anemia and hypoproteinemia continues, or localized abscsses fail to improve with chemotherapy, operative intervention is indicated. Complications of amebomas that require operative intervention include failure to respond to chemotherapy, perforation, hemorrhage, ulceration, stricture or fistula formation and obstruction. Anti-amebic chemotherapy can reduce intestinal complications so elective operative procedures should be delayed to allow time for adequate treatment.


[Hypoglycemic convulsions and hypopituitarism].

A case of hypopituitarism in a female, aged 6 months, whose first symptom consisted of hypoglycemic convulsions is reported. Making use of TRH we confirm the hypothalamic origin of pituitary failure. Substitution therapy controlled the episodes of convulsions and normalized the growth rate of the child.


Collagenase activity in epidermoid carcinoma of the oral cavity and larynx.

Summary--Tumor invasion requires the breadkdown of the main structural protein, collagen. A series of fourteen epidermoid carcinomas of the larynx and oral cavity produced a collagen dissolving enzyme in vitro as demonstrated by the breakdown of 14C-labeled collagen. Oral cavity tumors showed greater activity than laryngeal carcinomas while both sites were more active than uninvolved mucosa from the same patients. Tumor associated collagenase activity, in common with previously described collagenases, can only be demonstrated in vitro and requires protein synthesis. Maximum tumor collagenase occurred at 24 hours in vitro and then declined as compared with the maximum collagenase at 72 hours in vitro produced by oral cavity mucosa. The 14 patients in our series were ranked in order of the collagenase activity of their tumors. At 18 months after the diagnosis, four of the six patients with the most active tumors were dead of cancer and one patient was alive with persistent cancer. High collagenase activity may be a factor in the clinical aggressiveness of epidermoid carcinomas of the head and neck.


Primary oat-cell carcinoma of head and neck.

Summary--A case of primary oat-cell carcinoma of the larynx in a 68-year-old white female is described, bringing to a total of three the reported cases in the world literature. The tumor is highly malignant and notorious for the speed with which  it metastasizes. Treatment should be aggressive from the beginning. We believe that even for a localized tumor of this kind, a minimal procedure should be widefield laryngectomy with elective radical neck dissection even if clinically, nodes are not palpable. Since the tumor is highly radiosensitive, radiotherapy should be used postoperatively.


The biochemistry of collagen.

Collagen synthesis and subsequent extracellular stabilization and turnover are multi-step processes. The sequence of events in these processes is reviewed and correlated with collagen pathologies.


[Experimental effect of bruneomycin on the energy metabolism of liver tissue].

The effect of an antitumor antibiotic bruneomycin on the energy metabolism in the liver tissue was studied. Four hours after the drug administration the consumption of glycogen and glucose in the liver tissue increased because of glycogenolysis activation, which was evident from increased activity of prosphorylase, phosphofructokinase, hexokinase and summation glycolytic activity. 24 and 48 hours after the antibiotic administration the balance of consumption and resynthesis of phosphate macroergs in the liver tissue impaired, which was evident from decreased levels of ATP, impairement of conjugation of the processes of oxidation due to impairement of permeability and structural integrity of the mitochondrial membranes. Further decrease in the glycogen liver levels was mainly due to suppressed resynthesis of glycogen because of destructive-necrotic processes. Simultaneously the processes of glycolytic splitting of glucose decreased which was evident from decreased activity of the enzymes and summation glycolytic activity.


Erythropoietin production in renal tumors.

A review of the pertinent literature on the relation of erythropoietin production to the presence of renal neoplasm suggests that erythropoietin may be produced either directly by the tumor or indirectly by its physical effect on the adjoining normal renal tissue. The most commonly found tumors which are associated with elevated levels of serum and urinary erythropoietin are the hypernephromas. However, the presence of erythropoietin and an associated erythrocytosis even here occurs only relatively infrequently. Some studies have demonstrated the presence of erythropoietin activity in tumor tissue itself but erythropoietin has not been isolated from renal tumor tissue. In some patients with Wilms' tumor, erythropoietin blood levels may also be increased; however, erythrocytosis in these patients is not a characteristic feature. Other renal tumors rarely produce erythrocytosis and presumably no erythropoietin. Possible explanations for the production of erythropoietin by renal tumors are discussed.


Interpretation of the tube coagulase test for identification of Staphylococcus aureus.

The tube coagulase test is a valid means of identifying Staphylococcus auerus, provided that only a firm clot that does not move when the tube is tipped is considered a positive reaction. The widely promulgated interpretation that all degrees of clotting in coagulase plasma are a positive identification of S. auerus was disproved by the use of other tests such as anaerobic glucose fermentation, thermonuclease production, and lysostaphin sensitivity. It was found that the source of supply of the coagulase plasma is a factor in the occurrence of false-positive coagulase test results. The use of a mixture of pig and rabbit plasma in the tube coagulase test is also discussed.


Production of hydrogen cyanide by Pseudomonas fluorescens.

Two Pseudomonas fluorescens isolates were found to produce hydrogen cyanide when cultured on either Trypticase soy agar supplemented with 0.5% yeast extract or on irradiation-sterilized chicken.


Prediction of adult height from height, bone age, and occurrence of menarche, at ages 4 to 16 with allowance for midparent height.

Multiple regression equations for predicting the adult height of boys and girls from height and bone age at ages 4 and upwards are presented. There is a separate equation for each half year of chronological age; and for pre- and postmenarcheal girls at ages 11 to 14. These are based on longitudinal data from 116 boys and 95 girls of the Harpenden Growth Study and the London group of the International Children's Centre longitudinal study. The bone age used is the revised version of the Tanner-Whitehouse standards, omitting the score for carpal bones (RUS age, TW 2 system). Boys aged 4 to 12 are predicted in 95% of instances to within plus or minus 7 cm of true height, and at ages 13 and 14 to within plus or minus 6 cm. Girls ages 4 to 11 are predicted to within plus or minus 6 cm; premenarcheal girls aged 12 and 13 to within plus or minus 5 and plus or minus 4 cm, respectively; and postmenarcheal girls aged 12 and 13 to within plus or minus 4 and plus or minus 3 cm, respectively. Prediction can be somewhat imporved by allowing for midparent height. One-third of the amount that midparent height differs from mean midparent height is added or subtracted. An alternative system of equations which are based on initial classification by bone age rather than chronological age is given. These have about the same accuracy as the equations based on initial classification by chronological age, but allowance for bone age retardation is less. It is not clear which system is preferable. The equations probably apply to girls complaining of tall stature and boys or girls complaining of shortness and needing reassurance as to normality. In clearly pathological children, such as those with endocrinopathies, they do not apply.


Seminoma of the anterior mediastinum.

Primary pure cell seminoma of the mediastinum is a rare and potentially fatal lesion. Encroachment on or invasion of adjacent structures is common, as are distant metastases. The differentiation between pure cell seminomas and mixed germ cell tumors should be made since the extreme radiosensitivity of seminomas improves the prognosis. The presence of metastases from primary mediastinal seminomas, as with testicular germinomas, need not be a deterrent to long-term cure. Recommended therapy consists of total or subtotal removal, radiation therapy (2,500 to 3,500 rads delivered over 2 to 4 weeks), and, in some patients, chemotherapy. A patient who was successfully treated with subtotal removal and irradiation is presented.


Bronchial adenoma.

Thirty-three patients with bronchial adenoma have been treated in U.S. Army hospitals. These included 27 with carcinoid, 3 with mucoepidermoid carcinoma, and 3 with adenoid cystic carcinoma. No deaths could be directly related to the adenomas during an average follow-up of 14.5 years.


Excitation-contraction uncoupling. The effect of hyperosomolar glycerol solution and antrolene sodium on mammalian muscle in vitro.

In the rat hemidiaphragm, in vitro dantrolene sodium irreversibly reduced twitch tension up to 20% of the original value in 20 minutes of exposure. The phenomenon was dose related. Dantrolene sodium did not affect (1) resting membrane potential (IMP); (2) miniature endplate potential frequency of amplitude; or (3) endplate potential amplitude decrement of repetitively stimulated partially curarized muscle. A combination of hyperosmolar solutions of glycerol and dantrolene sodium abolished contraction of mammalian muscle with no demonstrable effect on neuromuscular transmission or muscle fiber RMP. With this technique, prolonged recording from repetitively activated muscle fibers is possible.


Histochemistry of trauma after electrode implantation and stimulation in the hippocampus.

Electrodes were implanted into the rat hippocampus in both hemispheres for increasing periods of up to 60 days, and the effects of trauma and electrical stimulation of enzymes controlling cell metabolism in the region of implantation were examined and assessed in relation to studies on humans. In the unstimulated hippocampus as a control, enzyme changes were mainly confined to a narrow area of tissue damage surrounding the electrode. The enzyme changes in response to trauma varied widely; some enzymes controlling tissue respiration showed early and rapid changes, increasing in hyperactive, swollen glial cells and vascular endothelium and decreasing in nerve cells and neuropile. Acid phosphatase activity also increased rapidly in glial cells; other phosphate-releasing enzymes increased more gradually with time. A turning point in these chages was apparent between 25 and 40 days, followed by a reversion to more normal levels at 60 days. Electrical stimulation of the hippocampus in the contralateral hemisphere produced no detectable enzyme changes from those of the unstimulated hippocampus.


Toxic polyneuropathy due to methyl n-butyl ketone. An industrial outbreak.

Cases of toxic distal polyneuropathy have been studied in a plant producing plastic-coated and color-printed fabrics. After the screenig of 1,157 employees, a total of 86 verified cases were detected. Of these, 11 were moderate to severe in intensity and usually with motor and sensory involvement; 38 were mild, with sensory signs prevailing; and 37 were minimal, but with characteristic electro-diagnostic abnormalities. Muscle weakness and electromyographic abnormalities were predominantly distal. Reflex loss was minimal. Sensory deficits were distal and limited to pain, touch, and temperature discrimination with occasional loss of vibration sense. The distribution of involvement severity of the disorder, and temporal course of the outbreak correlated with exposure with methyl n-butyl ketone. After elimination of this agent improvement was noted in the majority of cases.


Mechanism of colistin-induced neuromuscular depression.

The effect of a high concentration of the antibiotic colistin sulfate on neuromuscular transmission was examined by recording intracellularly from rat diaphragm blocked with tubocurarine chloride. The presynaptic effect of colistin was different from that of neomycin sulfate, which reduces the probability of acetylcholine release: Colistin reduced that the quantum content of the initial and the last 20 end-plate potentials of a train of 40 stimuli without altering the probability of acetylcholine release. The quantum size was reduced, while the frequency of miniature end-plate potentials was unchanged.


Adrenoleukodystrophy. Electron microscopic findings.

Ultrastructural and neurochemical studies were done on three male patients with adrenoleukodystrophy. In each case, the affected white matter contained enlarged glial cells filled with pathognomic intracytoplasmic inclusions consisting of electron-lucent spicules bounded by 25-Angstrom wide membranes. Similar inclusions were present in adrenocortical cells. These findings and a review of 47 reported cases indicate that adrenoleukodystrophy is a storage disorder caused by a sex-linked recessive error of metabolism.


Methysergide in the treatment of narcolepsy.

Five patients with narcolepsy (four with the allied symptom of cataplexy) were treated with the serotonin antagonist methysergide. All patients had as good control of their sleep attacks while on methysergide therapy as on a control period of dextroamphetamine therapy. The cataplexy was less well controlled by methysergide than by dextroamphetamine, but improved when compared to a period without medication. Two patients developed severe calf claudication while on methysergide.


[Antigenic characteristics of hamster cells transformed by polyoma virus].

A hamster cell line derived from a polyoma virus induced tumour (G-Py) was found positive for TSTA and S antigen detected by the surface immunofluorescence technique.  Lung metastases were observed in some hamster able to reject a G-Py cell challenge dose injected by intradermal route.


[Double clone in cells of pleural effusion caused by metastasis of oat cell carcinoma of the lung].

The Authors report a chromosome study carried out on pleural effusion cells from metastases of oat cell carcinoma.  They observed a double cell clone: the former, with 44 chromosomes, with various alterations of the karyotype and a big acrocentric "marker", the latter, with 45 chromosomes with various alterations of the karyotype and a big submetacentric "marker".  From these observations the Authors draw some interpretative considerations.


Neurotoxicity of acrylamide and its analogues and effects of these analogues and other agents on acrylamide neuropathy.

N-Hydroxymethylacrylamide, N-methylacrylamide, and N,N-diethylacrylamide produce peripheral neuropathy in rats. Seven other compounds related to acrylamide do not produce neuropathy. Rats given one of the three neurotoxic compounds are more susceptible to acrylamide. A regime for testing acrylamide analogues for neuro-toxicity is suggested. DDT, phenobarbitone, or high dietary concentrations of vitamin A or E have no effect on the development of acrylamide neuropathy in rats. Acrylamide produces neuropathy in hens but not in frogs or goldfish.


A cohort study of bronchial carcinomas in workers producing chromate pigments.

A cohort study of the incidence of bronchial cancer in male workers in a small company producing chromate pigments is presented. Altogether 133 workers had been employed by the company from the time production was started in 1948 until the end of 1972. Workers with more than three years employment were included in the study, and three cases of bronchial carcinoma were found among the 24 workers who fulfilled this requirement. Based on the data of the Cancer Registry of Norway the risk of bronchial cancer for a corresponding group of the general population was found to be 0.079, which gives a risk ratio for exposed workers of approximately 38. The average age of the cancer patients was as low as 50 years at the time of diagnosis. All workers in the company had been exposed mainly to zinc chromate dust, and the exposure levels of the workers developing bronchial cancers had probably been from 0-5 to 1-5 mg Cr/m-3 for six to nine years. Two of the three patients were smokers. It is assumed that exposure to chromate pigments, and probably to zinc chromate, may be related to the increased incidence of bronchial cancer in this group of workers. The possibility of a contributing effect of tobacco smoking in at least two of the three cases cannot be ruled out.


Long-term effects on the health of men engaged in the manufacture of tolylene di-isocyanate.

A prospective study of workers engaged in the manufacture of tolylene di-isocyanate (TDI) was carried out over nine years. The study was in two parts: (1) assessment of the symptoms and ventilatory function of men working on the plants who did not complain of respiratory symptoms; (2) assessment of the long-term effects in men who were removed from the plants because of respiratory symptoms. Use of the MRC Questionnaire on Respiratory Symptoms in a group of 76 TDI workers showed no significant differences in symptoms compared with 76 control subjects. Annual lung function tests were carried out on 180 asymptomatic men with possible exposure to TDI. The relationship of the forced expiratory volume in 1 second (FEV1) and forced vital capacity (FVC) to the height, age, and duration of exposure was examined by linear regression analysis. The equations formulated did not differ significantly from those calculated for non-TDI exposed subjects. Thus exposure to very small concentrations of TDI does not necessarily cause symptoms or a drop in ventilatory capacity. Forty-six TDI workers who had developed symptoms reported more long-term symptoms than a comparable number of controls. The difference was significant at the 1% level. Ventilation test records of 61 TDI workers who had developed symptoms were available since it was possible to use the records of men who had left the company. The lung function of this group was matched against predicted figures obtained from a control group of 608 subjects living in the same area as the exposed group but not exposed to TDI. It was found that the FEV1 of the affected men was on average 267 ml lower than predicted, and the FVC 269 ml lower. These findings are confirmed by examination of the indivdual records of a futher 20 men.


Studies of the viral flora in keratoconjunctivitis sicca.

Fifty patients suffering from KCS and exhibiting various patterns of corneal staining with fluorescein were examined for adenovirus types 3, 7, 8, and 14, herpes simplex, and the TRIC group by tissue culture and immunofluorescein techniques. No viruses were detected by either methods, and it is considered that presumption of virus involvement in these cases is untenable.


Dermatofibrosarcoma protuberans in the parotid gland--a case report.

A recurrent dermatofibrosarcoma protuberans (storiform fibrocytic histiocytoma) in the parotid gland of a 50-year-old Nigerian male is reported. Recent knowledge of the histiogenetic origin of the tumor is mentioned.


Interaction of reduced nicotinamide adenine dinucleotide with beef heart s-malate dehydrogenase.

The interaction of NADH with s-malate dehydrogenase isolated from beef heart was studied in 20 mM potassium phosphate (pH 6.9)-1 mM EDTA, with forced dialysis, fluorescence, and temperature-jump techniques. Measurements of the change in fluorescence of NADH when it is titrated with enzyme indicate NADH bound to monomeric and dimeric enzyme have different fluorescence yields. These data and the results of direct binding studies can be explained in terms of a model in which the NADH binding sites on dimeric enzyme are equivalent or nearly equivalent, and NADH binding to monomeric enzyme occurs with an affinity very similar to that of the dimer. However, the fluorescence enhancement of NADH on binding to the enzyme is different for the monomer and for each of the two dimer sites.


Conformational flexibility of angiotensin II. A carbon-13 spin-lattice relaxation study.

Carbon-13 spin-lattice relaxation times (T1) have been determined for the carbon in the octapeptide hormone [5-isoleucine]-angiotensin II in aqueous solution. Two possible models for molecular motion are considered: isotropic overall motion of the hormone with internal motion of some residues and anisotropic overall molecular motion. The data are interpreted in detail using the former model. The alpha carbons of the peptide backbone are all equally restricted in their motion. The correlation time for overall molecular reorientation, calculated from an everage T1 value of 95 msec for the alpha carbons in the peptide backbone, is ca. 5 times 10-10 sec. The carbons in the side chains are more mobile than those in the peptide backbone, with the exception of the side chain of the Tyr residue which does not undergo rapid segmental motion. We propose that [5-isoleucine]-angiotensin II has a restricted backbone conformation and that the alpha carbons of the N- and C-terminal residues are constrained to nearly the same extent as the remaining alpha carbons in the peptide backbone. Chemical shift data indicate that the Pro residue adopts the trans conformation about the His-Pro bond and that the imidazole ring of His has a strong preference for the N-tau -H tautomer.


Purification of the sex steroid binding protein from human serum.

The sex steroid binding protein from human pregnancy serum was purified to homogeneity by affinity chromatography and preparative polyacrylamide gel electrophoresis. The selective adsorbants were prepared by coupling [3H]-5alpha-dihydrotestosterone 17beta-hemisuccinate to 3,3'-diaminodipropylamine-agarose, poly(Lys-DLAla)-agarose, and albumin-agarose. The most effective adsorbant purifying for the binding protein was 5alpha-dihydrotestosterone 17beta-hemisuccinyl-3,3'-diaminodipropylamine-agarose. A preparative procedure with 5alpha-dihydrotestosterone 17beta-hemisuccinyl-3,3'-diaminodipropylamine-agarose yielded active material which was further purified by preparative polyacrylamide electrophoresis at pH 9.5. Homogeneity was shown by analytical disc gel electrophoresis at three different pH units. A single radioactive band corresponding to the stained band was shown by incubating with [1,2-3H]-5alpha-dihydrotestosterone prior to electrophoresis. The radioactive peak corresponding to the pure sex steroid binding protein could not be detected when a 100-fold excess of 17beta-estradiol was present in the incubation prior to electrophoresis demonstrating the specific sex steroid binding properties of this protein. The migration of this peak was identical with that obtained when diluted serum was electrophoresed under the same conditions in the presence of [1,2-3H]-5alpha-dihydrotestosterone indicating that no significant changes in the molecular characteristics of the binding protein occurred during the purification procedure. The presence of carbohydrate in the pure protein was shown by the periodic acid-Schiff reagent procedure. Selective adsorbants containing 17beta-estradiol linked at the 3 position were ineffective in retaining sex steroid binding protein activity.


Studies on interaction between histone V (f2c) and deoxyribonucleic acids.

Histone V (2fc) from chick erythroctes was used in the study of its interaction with DNA from various sources. Complexes between this histone and DNA were formed using the procedure of continuous NaCl gradient dialysis in urea. Two physical methods, namely thermal denaturation and circular dichroism (CD), were used as analytical tools. Thermal denaturation of nucleohistone V with chick or calf thymus DNA shows three melting bands: band I at 45-50 degrees corresponds to free base pairs; band II at 75-79 degrees, and band III at 90-93 degrees correspond to histone-bound base pairs. In histone-bound regions, there are 1.5 amino acid residues/nucleotide in nucleohistone V. In contrast, a value between 2.9 and 3.3 was determined for nucleohistone I (fl) (H. J. Li (1973), Biopolymers 12, 287). Similar melting properties have been observed for histone V complexed with bacterial DNA from Micrococcus luteus. Histone V binding to DNA induces a slight transition from a B-type CD spectrum to a C-type spectrum. Trypsin treatment of nucleohistone V reduces melting band III much more effectively than band II. Such a treatment also restores DNA to B conformation in the free state. Reduction of the melting bands of nucleohistone V by polylysine binding follows the order of I greater than II greater than III, accompanied by the increase of a new band at 100 degrees. When two bacterial DNAs of varied A + T (adenine + thymine) content simultaneously compete for the binding of histone V, the more (A " T)-rich DNA is selectively favored. Under experimental conditions described here, Clostridium perfringens DNA with 69% A + T is bound by histone V in preference to chicken DNA with 56% A + T although the latter has natural sequences for histone V binding.


The assay and isolation of DNA rings using an ATP-dependent endonuclease.

The ATP-dependent endonuclease from Hemophilus influenzae is relatively inactive on closed or open DNA rings, yet rapidly hydrolyzes single- or double-chained linear DNA. This enzyme in combination with an exonuclease (exo VII) has been shown to spare various circular DNA molecules including those having single-chain regions of significant length. However, rings containing single-chained regions are broken at a rate depending on the length of these regions. By admixing a linear DNA of alternate radiolabel, a simple assay for DNA rings has been developed. The application of this procedure to the assay of folded rings from Drosophila DNA is demonstrated.


Partial reconstitution of active ribosomes and 50S subunits.

Escherichia coli ribosomes and their 50S subunits disassembled by LiCl treatment can be reconstituted into structurally completed but inactive particles. However, peptidyltransferase and polyphenylalanine synthesizing activity can be partly recovered by the addition of methanol to the reconstitution system. Furthermore, entirely active ribosomes and 50S subunits are reconstituted when methanol is present during the initial treatment with LiCl to disassemble the ribosomal components. The presence of methanol (10% v/v) during this treatment diminished the release of some proteins but does not affect the separation of the 5S RNA.


Structures of gangliosides from bovine adrenal medulla.

Five gangliosides, accounting for over 95% of the total ganglioside fraction, were isolated from bovine adrenal medulla by preparative thin-layer chromatography and the carbohydrate structures determined by a combination of periodate oxidation and permethylation techniques. Partially methylated alditol acetates were generated from the neutral sugars of the fully methylated glycolipids and identified by gas-liquid chromatography. Substitution on N-acetylgalactosamine was determined by methanolysis of the permethylated ganglioside, acetylation of the products, and identification of the resulting substituted methyl glycosides by GLC. Periodate oxidation followed by borohydride reduction confirmed some of the linkages and demonstrated the absence of (2-8) linkages between sialic acid units. Mass spectrometry of the permethylated gangliosides gave information on sugar sequence at the nonreducing end.


Structure-function relationships in glucagon: properties of highly purified des-His-1-, monoiodo-, and (des-Asn-28, Thr-29)(homoserine lactone-27)-glucagon.

We have compared the ability of glucagon and three highly purified derivatives of the hormone to activate hepatic adenylate cyclase (an expression of biological activity of the hormone) and to compete with [125]glucagon for binding to sites specific for glucagon in hepatic plasma membranes. Relative to that of glucagon, biological activity and affinity of [des-Asn-28,Thr-29](homoserine lactone-27)-glucagon, prepared by CNBr treatment of glucagon, were reduced equally by 40- to 50-fold. By contrast, des-His-1-glucagon, prepared by an insoluble Edman reagent and highly purified (less than 0.5% contamination with native glucagon), displayed a 15-fold decrease in affinity but a 50-fold decrease in biological activity relative to that of the native hormone. At maximal stimulating concentrations, des-His-1-glucagon yielded 70% of the activity given by saturating concentrations of glucagon. Thus, des-His-1-glucagon can be classified as a partial weak agonist. Highly purified monoiodoglucagon and native glucagon displayed identical biological activity and affinity for the binding sites. Our findings suggest that the hydrophilic residues at the terminus of the carboxy region of glucagon are involved in the process of recognition at the glucagon receptor but do not participate in the sequence of events leading to activation of adenylate cyclase. The amino-terminal histidyl residue in glucagon plays an important but not obligatory role in the expression of hormone action and contributes to a significant extent in the recognition process.


The mechanism of microsomal and mitochondrial nitroreductase. Electron spin resonance evidence for nitroaromatic free radical intermediates.

Electron spin resonance spectra are observed during the enzymatic reduction of many nitrophenyl derivatives by rat hepatic microsomes or mitochondria. The spectra indicate that nitroaromatic anion radicals are present and are freely rotating in aqueous solution at a steady-state concentration of 0.1-6 muM. The rate of formation of p-nitrobenzoate (NBZO) dianion radical in microsomal incubates is consistent with the radical being an obligate intermediate in the reduction of NBZO to p-aminobenzoic acid. A model system consisting of NBZO, NADPH, and FMN, but no heme-containing compounds, also reduced NBZO to the NBZO dianion free radical. The steady-state concentration of the anion radicals in microsomal systems is not altered by CO. This observation, together with the results from the model system, suggests that the formation of nitroaromatic anion radicals is mediated through a flavine and not cytochrome P-450. The oxidation of the anion radical intermediate by O2 to the parent nitro compound is proposed to account for the well-known O2 inhibition of microsomal nitroreductase.