[Lymphocyte transformation test in the calf].

Studies into the effects of the environment on health and performance of agricultural animals are of clear-cut relevance to farming practice. The theoretical and practical importance of lymphocyte transformation to any assessment of immunological reactivity under defined environmental conditions, therefore, is reported in this paper. Communicated is methodical experience obtained from isolation of lymphocytes from peripheral blood of calves, lymphocyte culturing, and morphological evaluation of lymphocyte transformation. Views are given on the suitability of the lymphocyte transformation test. The following results have been obtained: Centrifugation of defibrinated blood, using "Visotrast-370", is recommended for lymphocyte isolation. Morphological evaluation will ensure a high degree of precision when 1 x 10(6) cells in one millilitre culturing fluid are to be cultured over three days, the culturing fluid consisting of 20 per cent of autologous serum and 80 per cent of Eagle medium, antibiotics, and a portion of 1-vol.% of phytohaemagglutinin (Wellcome). The lymphocyte transformation test, for its methodical accuracy, is good enough to detect and identify environmental effects on immunological reactivity of calf. It is likely to reflect the immunological response to an antigen application.


Differentiating exogenous psychiatric illness from schizophrenia.

Retaining an individual on psychiatric dispensary lists long after a single psychotic episode can result in unnecessary restriction of his or her social-vocational rights and responsibilities. This study demonstrates that an early clinical differention can be made between exogenous psychoses and progressive schizophrenia. The validity of the clinical differentiation was enhanced by demonstrating that a computer learning and pattern-recognition program was capable of using signs and symptoms recorded in the first psychotic episode to make a differential diagnosis that coincided closely with diagnoses made at a later date by clinicians aware of the subsequent clinical course. This kind of approach to standardized nosologic principles may expand the possibility for more appropriate application of psychotropic medications, psychotherapy, and somatic treatments, as well as more accurate social-vocational prognoses.


Localization of IgE in tissues by an immunoperoxidase technique.

An indirect immunoperoxidase technique was used to study the content and distribution of IgE in formaldehyde solution-fixed, paraffin-embedded adenoid and nasal tissues of atopic and nonatopic persons. Adenoid tissue from 15 patients, and nasal tissue from five patients with symptoms of inhalant allergies and augmented serum levels of total and allergen-specific IgE were examined with this method. Adenoid tissues from five patients without allergic symptoms were studied for comparison. A rich content of IgE, largely within the cytoplasm of the plasma cells, was observed in tissues of atopic subjects. The sections from nonallergic persons contained few weakly staining IgE-positive cells. These observations provide anatomic support for the role of IgE in hypersensitivity reaction of immediate type. This immunoperoxidase technique, circumventing the principal shortcomings of the immunofluorescence procedures, affords a highly sensitive and practical approach to cellular and tissue localization of IgE.


Histiocytic lymphoma in a patient with Lennert's lymphoma. Report of a case with unusual cytoplasmic inclusions.

The transformation of Lennert's lymphoma into histiocytic lymphoma appears to be part of the natural history of the disorder rather than therapeutically induced. Unusual paranuclear nonmembrane-bound filamentous inclusions were found in many of the neoplastic cells in the histiocytic lymphoma. We report evidence for the immunoglobulin nature of this material, indicating a B cell origin of the neoplasm.


Skeletal muscle in polymyositis. Immunohistochemical study.

Thirty-two patients with adult-onset polymyositis uncomplicated by cancer or systemic connective tissue disease were studied. Muscle biopsy specimens were examined with direct immunofluorescence microscopy and results were compared with those in 94 control subjects. Sarcolemmal and sarcoplasmic staining were observed in both groups and considered to be nonspecific. Immune deposits in the muscle microvasculature were present in some cases of systemic lupus erythematosus and dermatomyositis but were not present in polymyositis. Our data suggest that the finding of vascular immunofluorescence excludes the diagnosis of adult polymyositis and implies that the pathogenesis of this disease and other idiopathic inflammatory myopathies may differ.


Surgical management of traumatic injuries of the right colon. Twenty years of civilian experience.

Three hundred seven patients with right colon injuries were treated at Parkland Memorial Hospital, Dallas, between 1953 and 1973. These injuries were managed with loop colostomies, suture and cecostomy, primary repair, right colectomy, diverting colostomy, and appendectomy or cecostomy alone. The indications and results of each type of repair are analyzed separately. The mortality (3.21%), morbidity (22%), and hospital stay were not related to the modality of treatment, but were influenced by the injuring agent. Adequate treatment of civilian right colon injuries implies careful individualization to avoid serious septic complications and subsequent mortality.


Should the ruptured renal allograft be removed?

During a 16-month period when 93 renal transplants were performed, eight kidney graft ruptures were detected within 18 days of transplantation, without evidence of venous obstruction. Six grafts were removed at the time of an exploratory operation for rupture and only one showed signs of probable irreversible rejection when examined by microscopy. Two graft ruptures were repaired and one of these grafts has had good long-term function 22 months later. These observations suggest that if bleeding at the site of grafts has had good long-term function 22 months later. These observations suggest that if bleeding at the site of graft rupture can be securely controlled and if the conditions of the patient and of the graft are favorable except for the rupture, it may be possible to save more than one of eight grafts.


The chemical and biochemical reactivity of dichlorvos.

The chemical structure, reactivity and metabolic fate of the insecticide dichlorvos (2,2-dichlorovinyl dimethyl phosphate) are discussed in relation to the possible genotoxicity of this and other methyl phosphate triesters. Recent attempts to demonstrate the methylation of DNA following exposure of bacteria and animals to dichlorvos are reviewed. On the basis of comparative data relating mutagenesis to methylation reactions, it seems entirely appropriate to conclude that the mutagenicity of dichlorvos to bacteria is due solely to methylation of the bacterial DNA under the conditions of these tests. However, the methylation of mammalian DNA could not be demonstrated under realistic exposure conditions (when the alkylating mutagen methyl methanesulphonate afforded clearly measurable methylation). The failure to detect methylation by dichlorvos in vivo is attributed to the operation of highly efficient enzyme-catalysed biotransformations which rely largely on the phosphorylating reactivity of dichlorvos. The biotransformation pathways, characterised mostly in the rat, appear to be common also to pig, mouse, hamster, and man.


Comparative properties of the inclusion body proteins of the nucleopolyhedrosis viruses of Neodiprion sertifer and Lymantria dispar.

A comparison was made on the properties of the inclusion body proteins of two insect viruses: the nucleopolyhedrosis viruses of the European pine sawfly, Neodiprion sertifer, Geoffroy, and the gypsy moth, Lymantria dispar, Linnaeus. The inclusion body proteins were characterized by the following parameters: amino acid composition, polyacrylamide gel electrophoresis in the absence and presence of sodium dodecyl sulfate--mercaptoethanol, isoelectric focusing, and alkaline protease activity. The properties of the inclusion body proteins of the two viruses were similar in many respects, but clear differences were observed. A principal difference was the absence of alkaline protease activity associated with the inclusion body proteins of N. sertifer nucleopolyhedrosis virus.


Macromolecular structure of nuclear polyhedrosis virus genome.

DNA preparations from nuclear polyhedrosis virus (NPV) of Galleria mellonella L. (GmL) were fractionated in high ionic strength neutral sucrose gradient. This procedure allowed a separation of supercoiled infectious DNA molecules with contour length of 48--52 microns from infectious open ring DNA molecule, and noninfectious linear DNA molecules of the same size. In addition a heterogeneity of supercoiled DNA molecules was detected. Covalently closed DNA molecules did not contain protein or ribonucleotide ligands which could be digested by pronase or pancreatic RNase treatment. It is concluded from data on the infectivity of different molecular forms of DNA and reassociation kinetics studies, that the genome of GmL NPV is a unique ring nucleotide sequence with a molecular weight of about 90--100 X 10(6).


[Pressing problems of the intravital morphological diagnosis of pathological states of the endometrium].

The important problems of morphological diagnosis of the endometrium conditions by scrapings are dealt with. The necessary information in the features of the functional organization of the normal and pathological endometrium as well as the required histochemical and histoenzymatic methods of examination are presented. The principles of diagnosis of glandular hyperplasia and atypical glandular hyperplasia of the endometrium and of differential diagnosis from highly diffentiated variants of endometrium cancer are described. The influence of hormonal therapy on endometrial carcinomas is discussed. It is emphasized that pathologists should give due consideration to the clinical data. A sample of the form for sending of the endometrial scrapings into the laboratory is presented.


[Immunomorphological method of study with a peroxidase antibody label].

Among different variants of immunoperoxidase method preference is given to its indirect variant with labeling of pure antibody with peroxidase by a two-step procedure. Examples of the use of this method for the detection of serum proteins, tissue antigens, and autoantibody are presented. The best results are achieved in treatment of sections with pure antibody instead of antisera.


[Bactericidal action of formalin on the causative agent of plague].

Ten per cent neutral formalin in a ratio 1 : 20 was demonstrated to be useful as a desinfecting and fixing substance in pathohistological examinations of organs and tissues animals infected with the causative agent of plague. With test-objects' volume of 1.0 cm3 the optimal period of exposure providing a permanent bactericidal effect is 24 hours, with volumes of 1.5 and 2.0 cm3-2 days. The fixing effect of formalin on tissues is exerted in 3-4 days.


[Disseminated intravascular coagulation in the practice of the pathologist].

Disseminated intravascular coagulation (DIC) of blood develops as a result of a sharp increase in the release of thromboplastic substances. The mechanism of disseminated thrombosis is switched in at the level of the microcirculatory bed with defibrination of the peripheral blood and subsequent hemorrhages and bleedings. The causes of DIC development may include complications of pregnancy and delivery, different kinds of shock including endotoxin shock, hemorrhage, hemolysis. Histomorphological findings in DIC are as follows: hemorrhagic syndrome, fibrin thrombi in capillaries, arterioles and venules of the skin, kidneys, adrenals, hypophysis, gastrointestinal tract, lungs and other organs followed by necroses and hemorrhages in these organs. Clinically, DIC is manifested by symptoms of insufficiency of the affected organs (acute renal insufficiency, Waterhouse-Fridericksen syndrome, etc).


History of occlusive cerebrovascular disease I. from Wepfer to Moniz.

Occlusive cerebrovascular disease of carotid and vertebral arteries was described by Wepfer in 1658 and by Willis in 1665. Wepfer thought apoplexy resulted from occlusive disease of the vessels. Willis described collateral channels between these vessels, which prevented the development of stroke.


History of occlusive cerebrovascular disease. II. After Moniz, with special reference to surgical treatment.

Angiography and related procedures are discussed. Ophthalmodynamometry, recording of bruits and temporal pulse, and thermography have been mentioned. Transient ischemic attacks (TIAs) and some causes are described: embolic obstruction, followed by absorption of the clot or its removal in the blood, with symtomatic recovery; hemodynamic crisis resulting from lowering of systemic blood pressure by hemorrhage due to trauma, gastrointestinal bleeding, and other causes with recovery following blood replacement. Special types of occlusive disease include fibromuscular hyperplasia of the carotid, dissecting aneurysm of the aortic arch and branches, causing carotid and vertebral occlusions, moyamoya disease, and low profusion syndrome, generalized or unilateral. Surgical management includes reconstructive surgery and endarterectomy of extracranial vessels supplying the brain and extracranial-intracranial vascular anastomosis.


Bromocriptine treatment of spasmodic torticollis. A double-blind crossover study.

A double-blind crossover study (12 weeks and 12 weeks) was performed to evaluate the effects of bromocriptine on the symptoms and serum prolactin levels of 14 women with spasmodic torticollis. While the serum prolactin level decreased in response to bromocriptine, no difference was found between the effects of placebo and bromocriptine on the symptoms of spasmodic torticollis. Our findings suggest that dopaminergic neurotransmission is not involved in the pathogenesis of this syndrome.


Variation in corneal graft size related to trephine technique.

To determine the range of variation in corneal transplant tissue size induced by slight changes in trephination technique, four human corneas that had been stored in McCarey-Kaufman medium and 11 intact human globes were used. Endothelial surface trephination yielded a smaller corneal button than epithelial trephination. In the intact eye, the higher the intraocular pressure was, the larger the resulting corneal diameter. With use of a trephine with a central plunger, the higher the obturator was, the larger the diameter of the resulting tissue. Also, the duller the trephine was, the larger the diameter of the cut, due to difficulty in tissue penetration. Thus, slight variations can markedly affect the match in size between donor and recipient.


Retropharyngeal mediastinal pneumatocele. Report of a case.

We report the case of an infant with penetrating neck trauma followed by an upper respiratory tract obstruction. This case presented a diagnostic and management dilemma because of the development of an unusual retropharyngeal collection of air. In our review of the medical literature, we were unable to find any other case with a similar manifestation and clinical course.


Reconstruction after total laryngopharyngectomy.

After resection of a hypopharyngeal cancer, utilization of the remaining pharyngeal mucosa in restoring pharyngoesophageal continuity is the procedure of choice. This is possible in most patients. However, a few patients have cancers that involve the posterior pharyngeal wall and esophageal introitus. Resection in this group involves complete removal of a segment of the laryngopharynx and reconstruction with the use of tissue from the adjacent side of the neck, chest, or alimentary canal. Between 1965 and 1974, 44 patients required reconstruction of a pharyngeal defect with the use of adjacent skin flaps or visceral interposition. Pharyngeal reconstruction with the use of these techniques involved multiple operative procedures, a greater morbidity, and a large time investment by the patient and surgeon. Survival of these patients was shorter than survival of patients who had repair by primary closure of the pharyngeal mucosa because of the extent of the primary lesions.


Ernst Reissner 1824--1878.

Ernst Reissner studied the formation of the inner ear initially using the embryos of fowls, then the embryos of mammals, mainly cows and pigs, and to a less extent the embryos of man. The embryos were placed in water for maceration and fixed in vinegar or black wax. Black wax also provided contrast and supportive blocks to enable the cutting of thin sections. Only the surface of the specimens could be studied after staining as they could not have been thin enough. By sectioning embryos at different stages, he was not only able to determine individual stages of the formation of the labyrinth and postulate its formation in man; but also to conclude that the web (the membrane) he noted in embryos is a permanent structure of the labyrinth in man.


A method for the assay of ablastin in the serum of rats infected with Trypanosoma lewisi.

This paper describes a simple quantitative assay for albastin, a factor present in the serum from rats infected with Trypanosoma lewisi, which prevents the division of the parasite. The assay measures in vitro the inhibition of the incorporation of 3H-TdR into the DNA of T. lewisi in the presence of serum from infected animals. Utilising this method, one can measure the titre of albastin in a particular serum sample and the time and duration of its appearance in the circulation of infected rats.


Morphological and functional interrelationships of bone cells and matrix.

1. Bone is unique in its combination of cells, matrix and mineral. 2. It has a dual function in skeletal and mineral homeostasis. 3. Cells which are functionally different act as coordinated units depending on a variety of signals which may be hormonal, ionic, mechanical and, possibly, electrical. 4. The coordinated activity of the cells is brought about by their having an intimate structural interrelationship, by the probability of their being subject to the influence of factors contained within an extra-cellular fluid circulating around all the cells underlying the mesenchymal cell envelope, and by the possibility that the osteoclasts and osteoblasts do not represent an end-stage in differentiation. 5. The understanding of the factors which normally control cellular differentiation and function may lead to methods of reversing bone loss, rectifying the effects of metabolic bone disease and promoting the healing of fractures.


Limitation of myocardial infarct size: review of experimental basis and clinical trials.

Limitation of myocardial injury after coronary occlusion is an exciting, recently recognised, but as yet unconfirmed, possibility in the treatment of myocardial infarction. Review of the experimental basis for the concept indicates that although manipulation of the ST segment of the electrocardiogram is not sufficient proof that myocardial necrosis can be limited, more robust experimental methods have confirmed the efficacy of a variety of interventions in experimental coronary occlusion. The effect of myocardial infarct size in determining acute prognosis and complications has been confirmed in clinical studies though the importance of infarct size lessens after recovery. Reported attempts at limiting myocardial necrosis in patients with myocardial infarction have so far not been sufficiently convincing to merit universal clinical application. Results of large scale randomised trials currently underway are awaited with interest.


Alcohol consumption and hypertension--the evidence from hazardous drinking and alcoholic populations.

To provide an enhanced perspective to observations of an association between moderate alcohol consumption and hypertension, information on the frequency of hypertension in heavy drinking and alcoholic populations was reviewed. Hypertension was found more often in ambulatory heavy drinkers and alcoholics than in more moderate drinkers and abstainers. This hypertensive state appeared to be at least partially reversible upon cessation of drinking. Additional investigation is needed to define the level of consumption associated with an increased risk of hypertension, the dose response pattern, and the determinants of irreversibility. Alcohol-related hypertension may constitute a risk factor for cardiovascular disease, thus contributing to the excess mortality experienced by heavy drinking populations. In view of the steady increase in per capita alcohol consumption which characterizes many affluent societies further clarification of this relationship is an urgent public health research priority.


Effect of metabolic conditions on protein turnover in yeast.

1. In yeast growing on ethanol a turnover rate of up to 2%/h was measured. As much as 80% of the protein was subject to turnover, and no marked heterogeneity in the rate of degradation of protein was observed. When the yeast grew on glucose, the protein was degraded at a lower rate (0.5-1%/h). 2. Starvation for a nitrogen source increased the rate of protein degradation severalfold, whereas deprivation of phosphate had only a marginal effect (30% increase). Removal of glucose from a medium containing 50mM-phosphate did not cause marked changes in the rate of protein degradation. In contrast, when the media were low in phosphate (0.1 mM) removal of glucose increased the rate of turnover 2-4-fold. 3. Protein degradation proceeded unimpaired when the intracellular concentration of ATP decreased from 4 to 1 mM, but stopped completely when it decreased below 0.3 mM.


IgG antibodies to double-stranded DNA in systemic lupus erythematosus sera. Independent variation of complement fixing activity and total antibody content.

Antibodies to double-stranded deoxyribonucleic acid were studied using the kinetoplast of Crithidia luciliae. Titers were determined separately by conventional immunofluorescence and the complement fluorescent technique, and results by the two methods were compared. Complement fixing activity varied independently of antibody content in whole serum and in IgG fractions. The well established correlation of complement fixing activity of this antibody with activity of lupus nephritis appears related, therefore, to qualitative rather than solely quantitative differences. This finding has important implications for the clinical assessment of patients with lupus, and investigations on the relationship of anti-DNA antibodies to lupus nephritis.


Ia-Type alloantigens and humoral autoimmune responsiveness in insulin-dependent diabetes mellitus.

In the search for markers either closely linked to or identical with the hypothetical "diabetogenic major histocompatibility gene," immune region-associated alloantigens were defined in 80 patients with insulin-dependent diabetes and in 107 controls. A close association between the Ia-type alloantigen DRw3 and DRw4 and insulin-dependent diabetes was obtained. DRw3 was found in 36% and DRw4 in 32% of the patients compared to 11% and 16%, respectively, of the controls. In addition, a significant influence of DRw3 and humoral anti-islet-cell autoimmunity could be observed, which was found to be due to a high incidence of DRw3 in those patients with islet cell antibody persistence. Islet-cell antibodies (ICA) were observed in 60% of the DRw3-positive patients compared to only 9% of the DRw3-negatives with longstanding disease (greater than 5 yr). These data show a significant association between insulin-dependent diabetes and the Ia-type alloantigens DRw3 (p uncorr. less than 0.0005) and DRw4 (p uncorr. less than 0.025). Furthermore, they provide direct evidence of an association between an Ia-type alloantigen and persisting humoral autoimmune responsiveness in man.


The role of glucagon in the regulation of plasma lipids.

The role of glucagon in regulating plasma lipid concentrations (nonesterified fatty acids, ketone bodies, and triglycerides) is reviewed. The effects of glucagon-induced insulin secretion upon this lipid regulation are discussed that may resolve conflicting reports in the literature are resolved. In addition, the unresolved problem concerning the pharmacologic versus physiologic effects of glucagon is stressed. Glucagon's role in stimulating lipolysis at the adipocyte serves two important functions. First, it provides plasma nonesterified fatty acids for energy metabolism and secondly, it ensures substrate for hepatic ketogenesis. In vitro, glucagon's lipolytic activity has been consistently observed, but in vivo, this activity has sometimes been obscured by the effects of glucagon-induced insulin secretion. Frequently, a biphasic response has been reported in which a direct lipolytic response is followed by a glucagon-induced insulin suppression of plasma nonesterified fatty acid concentration. When the glucagon-induced insulin secretion has been controlled by various in vivo techniques, glucagon's lipolytic activity in vivo has frequently been demonstrable. In the 1960s, in vitro liver perfusion experiments demonstrated that glucagon enhanced hepatic ketogenesis independent of glucagon's lipolytic activity. However, this direct effect of glucagon on the hepatocyte was not universally accepted because of conflicting reports in the literature. Failure to observe an in vitro ketogenic effect of the hormone in some studies may have been due to suboptimal experimental conditions. Certain factors are now known to influence the ketogenic response, such as the concentration of fatty acids in the media and the nutritional status of the animal. Under optimal in vitro conditions with liver preparations from fed animals, the ketogenic response to physiologic concentrations of glucagon has been demonstrated. However, further study is necessary to define the quantitative ketogenic role of the hormone. In spite of this early in vitro work, glucagon was not definitely shown to be ketogenic in vivo (independent of fatty acid availability) both in the rat and in diabetic man until 1975. Since these observations, several reports have confirmed the ketogenic action of glucagon in vivo by direct hepatic catheterization experiments. Glucagon's role in decreasing hepatic triglyceride synthesis and secretion in vitro has been repeatedly shown but the mechanism is unresolved. This lipid regulatory action of glucagon has been more difficult to demonstrate in vivo because of the many variables that affect triglyceride synthesis. Under specific experimental conditions, however, glucagon has been shown to decrease plasma triglyceride concentration in man at both physiologic and pharmacologic concentrations. Hepatic catheterization experiments have also confirmed this effect in man. The regulation of lipids by glucagon fits well into its role as a stress hormone...


Characterization of dermal collagen in systemic sclerosis.

The amount of dermal collagen is increased in systemic sclerosis. However, unlike certain inflammatory conditions, the relative proportions of Type I and Type III collagens are closely similar to those found in normal adult dermis. Similarly, no change in the distribution of the collagen types could be detected by immunofluorescent staining, although a considerable thickening of the epidermis was clearly evident in all the sclerotic lesions examined.


Herpes gestationis: immunopathological and ultrastructural studies.

Eleven patients with the clinical picture of herpes gestationis were investigated. Biopsies were taken from involved and uninvolved areas of skin and the immunopathological and microscopic changes studied. Direct immunofluorescence showed a deposition of C3 and/or IgG at the basement membrane zone in the involved skin of nine patients and the uninvolved skin of five. Immuno-electron microscopy using a multistep peroxidase antiperoxidase method revealed the in vivo deposition of IgG at the basal plasma cell membrane that extended into the lamina lucida. Light microscopy of urticarial and vesicular lesions showed a marked oedema of the papillary dermis with an inflammatory cell infiltrate that was mainly perivascular. There was spongiosis of the epidermis with oedema and necrosis of the basal cells and in several specimens sub-epidermal clefts with bulla formation. Electron microscopy confirmed the marked degenerative and necrotic changes of the basal cells in the involved areas of skin.


Melanocyte mutation in halo naevus and malignant melanoma?

Halo naevus cells on frozen sections react with sera from patients with benign halo naevus or with malignant melanoma. Halo naevus cells also react with our specifically absorbed heterologous antihuman malignant melanoma antiserum (RAMMA). Since the cytoplasm of all the naevus cells within the halo naevus react with sera from patients with malignant melanoma, this implies that the entire population of melanocytic cells in the mole has undergone mutation, because non-pathological skin melanocytes do not react.


Coexistence of bullous pemphigoid and pemphigus foliaceus.

A patient with bullous pemphigoid developed pemphigus foliaceus 5 years later. Mixed bullous diseases are rare and this is the first reported case of coexisting bullous pemphigoid and pemphigus foliaceus.


Three dimensional microscopic surface profiles of membranes reconstructed from freeze etching electrol micrographs.

A method of three-dimensional reconstruction of the surface profile of artificial and natural membranes from freeze quenched electron micrographs is presented. The method is based on the analysis of the variation in thickness of platinum layers, deposited under an oblique angle. In essence, it is reminiscent of the method of Eratosthenes to measure the earth's radius. The thickness of etch-like protrusions of membranes could be determined to an accuracy of about 3 A. True distances on curved surfaces rather than projections of distances are obtained. The method has been applied to both model membranes and biological membranes. The essential results are: 1. Detailed information on the symmetry and the molecular structure of the crystalline phases of dimyristoyl phosphatidylcholine was obtained. The microscopic surface profile of the ripple structure observed between the pretransition and the main transition was analysed. In accordance with a previous model we found that the ripple structure is caused by the spontaneous curvature of the monolayers. The surface profiles of the ripple structure and of the low temperature biaxial phase could be clearly distinguished. 2. The sizes and shapes of lipid domains formed by both thermically and charge-induced lateral phase separation were determined. This showed that the visual inspection of electron micrographs may lead to a considerable underestimation of the domain size. Conclusions may be drawn concerning the different phases formed upon lateral phase separation. 3. As a biological example, yeast cell membranes were studied. The method allows one to distinguish between different membrane-bound proteins by measuring the width-to-height ratio of the particles. The deformation of the lipid layer in the environment of the proteins may be determined. This deformation contains information about lipid-mediated long-range interactions between membrane proteins.


Active K+ transport in Mycoplasma mycoides var. Capri. Net and unidirectional K+ movements.

Analysis of the cation composition of growing Mycoplasma mycoides var. Capri indicates that these organisms have a high intracellular K+ concentration (Ki: 200--300 mM) which greatly exceeds that of the growth medium, and a low Na+ concentration (Na+i: 20 mM). Unlike Na+i,K+i varies with cell aging. The K+ transport properties studied in washed organisms resuspended in buffered saline solution show that cells maintain a steady and large K+ concentration gradient across their membrane at the expense of metabolic energy mainly derived from glycolysis. In starved cells, K+i decreases and is partially compensated by a gain in Na+. This substitution completely reverses when metabolic substrate is added (K+ reaccumulation process). Kinetic analysis of K+ movement in cells with steady K+ level shows that most of K+ influx is mediated by an autologous K+-K+ exchange mechanism. On the other hand, during K+ reaccumulation by K+-depleted cells, a different mechanism (a K+ uptake mechanism) with higher transport capacity and affinity drives the net K+ influx. Both mechanisms are energy-dependent. Ouabain and anoxia have no effect on K+ transport mechanisms; in contrast, both processes are completely blocked by dicyclohexylcarbodiimide, an inhibitor of the Mg2+ -dependent ATPase activity.


Evidence for a magnesium- and ATP-dependent calcium extrusion pump in dog erythrocytes.

Intact dog erythrocytes, whose Ca2+ permeability had been increased with A23187 still maintained intracellular Ca2+ below electrochemical equilibrium indicating that they could extrude Ca2+. This extrusion required no Na+ gradient but apparently depended on intracellular ATP and Mg2+ suggesting that it was mediated by an ATP-fuelled Ca2+ pump.


The influence of temperature and incubation time on deformability of human erythrocytes.

Human erythrocytes have been heated and stressed in a novel and controlled manner using rectangular microcapillaries. Heated cells attached to the capillary wall were stressed by liquid flow. Under particular conditions of stress, temperature and incubation time the body of the cell could be pulled in the flow, retaining a connection with the glass by means of a narrow process or tether. The tethers appear as: regularly beaded, irregularly beaded or without beads depending upon the incubation conditions. We have outlined the incubation regimes necessary to achieve these different responses in the temperature range 48--55 degrees C. The cells become less deformable as the incubation is continued beyond an optimum time. The behaviour of the tether is compared with that of a viscoelastic liquid. Circular dichroism studies of ghost membranes show that the denaturation of membrane proteins is partially reversible when incubation times are similar to those required to bring about a loss of deformability.


The effect of differential methylation by Escherichia coli of plasmid DNA and phage T7 and lambda DNA on the cleavage by restriction endonuclease MboI from Moraxella bovis.

The nucleotide sequence recognized and cleaved by the restriction endonuclease MboI is 5' GATC and is identical to the central tetranucleotide of the restriction sites of BamHI and BglII. Experiments on the restriction of DNA from Escherichia coli dam and dam+ confirm the notion that GATC sequences are adenosyl-methylated by the dam function of E. coli and thereby are made refractory to cleavage by MboI. On the basis of this observation the degree of dam methylation of various DNAs was examined by cleavage with MboI and other restriction endonucleases. In plasmid DNA essentially all of the GATC sequences are methylated by the dam function. The DNA of phage lambda is only partially methylated, extended methylation is observed in the DNA of a substitution mutant of lambda, lambda gal8bio256, and in the lambda derived plasmid, lambdadv93, which is completely methylated. In contrast, phage T7 DNA is not methylated by dam. A suppression of dam methylation of T7 DNA appears to act only in cis dam. A suppression of dam methylation of T7 DNA appears to act only in cis since plasmid DNA replicated in a T7-infected cell is completely methylated. The results are discussed with respect to the participation of the dam methylase in different replication systems.


Fractionation and structural elucidation of the active components of aurintricarboxylic acid, a potent inhibitor of protein nucleic acid interactions.

Commercially available, as well as synthetically prepared, samples of aurintricarboxylic acid (a widely employed potent inhibitor of protein nucleic acid interactions) consist mostly of a heterogeneous collection of polymers, as shown by fractionation schemes utilizing both dialysis and ultrafiltration, and by molecular weight measurements. 13C-NMR studies suggest that the polymeric material is of the phenol-formaldehyde type; inhibitory assays that depend on the formation of a protein-nucleic acid complex revealed that potency varied directly with the molecular weight of the polymer. Fractions of molecular weight 400 were essentially inactive.


Specificity and selectivity of diacylglycerolphosphate synthesis in Escherichia coli.

The glycerolphosphate and 1-acylglycerolphosphate acyltransferase systems Escherichia coli membranes show relatively low specificities for acylcoenzymes A when maximal velocities for the respective acyl-coenzymes A are compared. However, the selectivities for palmitate and oleate in the acylations of the 1- and 2-positions of glycerolphosphate moiety, respectively, are higher at lower concentrations of acceptors in the presence of an equimolar mixture of palmitoyl-CoA and oleoyl-CoA. More 1-palmitoyl-2-oleoyl-glycerolphosphate species and less other species were synthesized at lower concentrations of glycerolphosphate. The fatty acyl moiety at the 1-position of 1-acylglycerolphosphate did not influence significantly the specificity for acyl-coenzymes A of the 1-acylglycerolphosphate acyltransferase system. Thus, the acceptor concentrations being kept low in vivo and in vitro are important for the highly selective incorporations of saturated and unsaturated fatty acids into the 1- and 2-positions of diacylglycerolphosphate, respectively, in the presence of mixtures of saturated and unsaturated acyl-coenzymes A while these acyltransferase systems exhibit relatively low specificies for acyl-coenzymes A when the respective maximal velocities are compared.


Immunochemical and immunohistochemical studies on the 27 S iodoprotein of dog thyroid with reference to thyroglobulin-like reaction of the parafollicular cells.

Our earlier finding that the thyroglobulin-like material responsible for the immunoreaction of parafollicular cells obtained in peak I fraction of Bio-Gel A-5m was followed up in the present study by an investigation of the immunochemical and immunohistochemical reactions of 27 S iodoprotein which was the most prominent material in the peak I fraction. The antibody was raised against completely purified 27 S iodoprotein which was obtained as follows: Thyroglobulin was extracted from dog thyroids and chromatographed initially on Bio-Gel A-5m and then on Bio-Gel A-50m. The area of 27 S migrated as a single bank on polyacrylamide gel slab electrophoresis. This was cut and eluted. Anti-27 S antiserum showed the same immunochemical patterns to 27 S and 19 S as anti-19 S antiserum with three different immunochemical methods: double diffusion test, one dimensional and two dimensional immunoelectrophoresis. The immunoperoxidase reactions of the anti-27 S antiserum and anti-19 S antiserum were restricted to follicular cells and luminal colloids. No reaction of the parafollicular cells was obtained by these antisera. Thus, 27 S iodoprotein shared common immunochemical and immunohistochemical properties with 19 S thyroglobulin. It was concluded that 27 S iodoprotein was not responsible for the thyroglobulin-like reaction of the parafollicular cells.


Comparative study between prokaryotes and eukaryotes by chemical iodination of ribosomal proteins.

Escherichia coli and Saccharomyces cerevisiae ribosomal proteins were chemically iodinated with 125I by chloramine T under conditions in which the proteins were denatured. The labelled proteins were subsequently separated by two-dimensional gel electrophoresis with an excess of untreated ribosomal proteins from the same species. The iodination did not change the electrophoretic mobility of the proteins as shown by the pattern of spots in the stained gel slabs and their autoradiography. The 125I radioactivity incorporated in the proteins was estimated by cutting out the gel spots from the two-dimensional electrophoresis gel slabs. The highest content of 125I was found in the ribosomal proteins L2, L11, L13, L20/S12, S4 and S9 from E. coli, and L2/L3, L4/L6/S7, L5, L19/L20, L22/S17, L29/S27, L35/L37 and S14/S15 from S. cerevisiae. Comparisons between the electrophoretic patterns of E. coli and S. cerevisiae ribosomal proteins were carried out by coelectrophoresis of labelled and unlabelled proteins from both species. E. coli ribosomal proteins L5, L11, L20, S2, S3 and S15/S16 were found to overlap with L15, L11/L16, L36/L37, S3, S10 and S33 from S. cerevisiae, respectively. Similar coelectrophoresis of E. coli 125I-labelled proteins with unlabelled rat liver and wheat germ ribosomal proteins showed the former to overlap with proteins L1, L11, L14, L16, L19, L20 and the latter with L2, L5, L6, L15, L17 from E. coli.


Alterations in the cell envelope composition of Proteus mirabilis during the development of swarmer cells.

Long, swarming cells of Proteus mirabilis had different proportions of some lipopolysaccharide components when compared to short cells, either agar grown or broth grown. Fluorescence spectrophotometry of antibody binding, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis indicated that the change was in the proportion of lipopolysaccharide with long O-antigenic sidechains, swarmer lipopolysaccharide relative to short sidechain lipopolysaccharide than the non-swarming cells. The proteins and phospholipids of the envelop remained the same during swarmer development. The results are discussed in relation to the increase in flagella synthesis and permeability to some antibacterial agents during swarmer development.


Epoxide hydrase in Trypanosoma cruzi epimastigotes.

1. Microsomal fractions from Trypanosoma cruzi epimastigotes catalyze the hydration of styrene oxide to styrene glycol. The activity is linear up to 45 min of incubation, is proportional to microsomal protein concentration within certain range, and has an optimum pH of 8.5. 2. Double-reciprocal plots indicate a Km value of 5.3 . 10(-4) M for styrene oxide and a V of 29.6 pmol of styrene glycol formed/min per mg protein at 37 degrees C. 4-Chlorophenyl-2,3-epoxypropyl either (Ki = 2.08 . 10(-4) M) and juvenile hormone I (Ki = 2.7 . 10(-4) M) are competitive inhibitors; whereas, 1-chloro-2,3-epoxypropane is a non-competitive inhibitor. The enzyme is induced about three-fold by 5 mM phenobarbital in the growth medium. 3. The epoxide hydrase is not activated by detergents but rather inhibited by concentrations of Tween-80 and Lubrol as low as 0.025%. 4. Experiments with intact cells indicate that about 3% of [8-14C]styrene oxide penetrates after 90 min of incubation; whereas, over 30% of juvenile hormone I is found intracellularly after the same incubation period. Intracellular styrene oxide is hydrated to styrene glycol to a significant extent and the in vivo hydration is increased by pretreatment with phenobarbital and inhibited upon the addition of 4-chlorophenyl-2,3-epoxypropyl ether. Only a small amount of the intracellular juvenile hormone I is recovered as the corresponding diol ester.


Intracellular binding of ethidium studied by photoaffinity labeling in vivo.

The azide analog of 14C-labeled ethidium bromide was mixed with yeast cells and when photolyzed by visible light, formed covalent complexes with all yeast cell organelles. The 14C counts were found in DNA, RNA and protein of yeast subcellular fractions, illustrating the complexity of binding of a drug which appears highly specific in its actions.


On the activation of the canine pepsinogens.

Acidification induces a conversion of canine pepsinogens by a sequential mechanism to the active pepsins. Activation in the presence of pepstatin, which strongly inhibits the pepsins but does not prevent the first step of activation, allows the isolation of the peptide released in this first step. This peptide inhibits the milk clotting activity of canine and also porcine pepsin. Canine pepsins obtained in the absence of pepstatin were characterized by amino acid composition, molecular weight, and activity against hemoglobin and milk and compared with those of other mammalian pepsins.


Stress analysis of single-tooth implants. II. Effect of implant root-length variation and pseudo-periodontal ligament incorporation.

This study delineates the effects of varying the root length as well as incorporating a soft pseudo periodonal ligament on the overall stress distribution for each of five different implant materials and one implant design.


Stress analysis of single-tooth implants. I. Effect of elastic parameters and geometry of implant.

Five different implant materials and three different geometries of ankylosed single-tooth implants were investigated. The stress distributions within and around the implants were determined by finite element analysis. Results show that the optimal combination of implant material and geometry can reduce implant and alveolar stresses by a factor of three when compared to an arbitrary implant design.


Some measurements of the shape and hydrodynamic properties of yeast phosphoglycerate kinase (E.C.2.7.2.3).

Using values obtained for sedimentation and diffusion constants the relative mass of phosphoglycerate kinase was calculated to be 45 800 +/- 1700. This value is higher than was previously estimated and the difference is thought to be caused by contamination of earlier crystalline preparations. Using the coordinates from X-ray crystallography it was found possible to calculate a frictional ratio for a linear dumb-bell (1.115) which compared well with the ratio calculated from diffusion (1.114 +/- 0.033). Since the calculated ratio for a bent molecule was 1.020 the natural state of the molecule in solution is essentially linear. From the concentration dependence of sedimentation and diffusion was calculated the effective interactive radius which resembles haemoglobin in its relationship to the molecular radius.


Method for continuous purification of biological material using immunosorbent.

A mechanical device for the continuous purification of biological material using immunosorbent was developed. The system consists of heat-sealed nylon pouches containing agarose-bound antibody, attached to an endless 35 mm wide Mylar belt that passes through four chambers sequentially. The biological material is bound and dissociated, and the immobilized antibody is regenerated for repeated isolation and purification of antigen. The belt design incorporates features to minimize carry-over between chambers and prevent damage to the agarose-bound antibody in repeated passes through the system. An existing batch method for the purification of human placental alkaline phosphatase using immobilized rabbit antisera was adapted to continuous purification in the device. The belt contained a low affinity immunosorbent and made five complete passes through the system. A decrease in antigen binding capacity between free immunosorbent suspensions and belt immunosorbent in pouches was observed. This was shown to be the result of the diffusion resistance offered by the pouch and the short exposure times of each pouch in the chambers. A decrease in antigen binding capacity between successive belt passes was also observed, and resulted from the inability of the agarose in the pouches to resuspend completely after each pass. The low efficiency of the agitation method and the roller device used to squeeze the pouches were the reasons for this deficiency.


[NAG infection in Rana temporaria with suppressed normal intestinal microflora].

Frogs of the Rana temporaria species with disturbed biocenosis induced by combined effects of hypothermia (+4 degrees C) and tetracycline were used as an experimental model. The animals were inoculated orally with cultured NAD-vibrios and subjected to clinical, bacteriological, immunomorphological and electron microscopical examinations. The lymph-blood system was shown to be a possible pathway for generalization of the infection. Several causes of long-term persistence of the vibrios in Rana temporaria are discussed.


[Immunological cross reactions between polysaccharides of Streptococci groups A and L].

Antibodies on sepharose immunosorbents containing A-polysaccharide-sepharose or synthetic beta-N-acetylglucosamine, were isolated from the sera of rabbits immunized with streptococci, group A, by means of affinity chromatography. Antibodies obtained from some sera with both immunosorbents reacted with streptococcus, group A and L polysaccharides. Partial identity of these polysaccharides was revealed by the immunodiffusion test. Absorption of antibodies with polysaccharides, group A and L, showed their different specificities. These antibodies could apparently be directed against the end parts of molecules of streptococcus, group A polysaccharide.


[Effect of mouse antiserum against isologous aggregated immunoglobulins on the transplantation and antitumor immunity].

It was shown that mouse antiserum against isologous thermically aggregated immunoglobulins MAAS injected repeatedly to the animals prolonged the survival of the skin allotransplant, induced Moloney's sarcoma diminished the latent period of the tumour development sharply increased the formation of the tumours which in the majority of cases, led to the death of mice, deranged the normal age barrier in fibrosarcoma induction, and sharply inhibited the intensity of Rauscher's leukemia development. This action of MAAS on the transplantation and antitumour immunity was of immunological nature.


[Effect of formaldehyde and its aminomethylol derivatives on E. coli strains with various defects of the DNA repair systems].

It was shown that aminomethylol compounds formed during reaction of formaldehyde with amino acids and formaldehyde as well exert a pronounced lethal action on E. coli strains with various defects of the DNA repair systems. The correlation between the extent of the DNA depurination caused by in vitro action of diverse aminomethylol derivatives and the inactivation of bacteria by these derivatives is revealed. The data obtained suggest that the inactivating effect of formaldehyde and its aminomethylol derivatives seems likely to be due to the formation of depurinized groups in bacterial DNA rather than to dimerization of purine bases.


[Immunoreactive luliberin in the visceral organs of rats].

The distribution of luliberin (luteinizing hormone-releasing hormone, LH-RH) was studied. LH-RH-like factor showing analogous immunochemical and chromatographic properties was detected in the liver, kidneys, duodenum, pancreas, adrenal glands and heart of rats. The concentration of immunoreactive LH-RH in the liver, kidneys, duodenum, pancreas and adrenal glands was nearly equal (5 to 7 pg per mg of methanol extract obtained from acetic acid extract of acetone powder), its concentration in the heart being somewhat lower (2 pg per mg of extract). Only minute amounts of this factor were present in blood cells. Immunoreactive LH-RH found in visceral organs may be of hypothalamic origin or may be synthesized in these organs (in situ).


Smoking and lung cancer: a review.

Cigarette smoking is the most important known factor in the causation of lung cancer. The evidence supporting this statement is enormous and irrefutable. Efforts to reduce and eventually eliminate cigarette smoking throughout the world should be strongly endorsed by all nations. The task will be difficult, but it can and must be accomplished.


Anti-aggregatory effect of prostacyclin (PGI2) in vivo.

Prostacyclin (PGI2) when infused intravenously reduced the mortality of rabbits given high intravenous doses of arachidonic acid (AA). Prostaglandins E1 and D2 were ineffective. Indomethacin pretreatment abolished the toxic AA effect. Since the lethal effect of AA is partly due to the formation of platelet aggregates it is concluded that PGI2 is a most potent anti-aggregatory prostaglandin in vivo.


Comparison of brush cytology before or after biopsy for diagnosis of gastric carcinoma.

A randomized trial has compared the results of brush cytology before or after biopsy in patients with solitary gastric lesions. One hundred and eleven carcinomas were identified in 324 patients. False positive histological reports were recorded in 2 patients but there were no false positive cytology results. The cumulative accuracy of biopsy and cytology in patients with carcinoma was 97 per cent, which was significantly better than biopsy alone (83 per cent, P less than 0.001) or cytology alone (87 per cent, P less than 0.001). The cumulative results of brushing before biopsy were significantly better than results obtained by brushing after biopsy (P less than 0.05) and indicate that brush cytology should normally be performed before biopsy.


Abdominal wound closure--the Jenkins's technique: a registrar's experience.

Jenkins's technique has been used to close 238 abdominal wounds. No dehiscence occurred in thie series. It is suggested that this method, which gives strict guidelines on wound closure, is ideal for teaching good technique for wound closure.


A comparison between disposable and non-disposable suction drainage units: a report of a controlled trial.

A randomized trial of high vacuum Redivac drains and low vacuum Portovac drains has been conducted in 46 female patients undergoing simple mastectomy through a transverse skin incision. The Redivac units drained less fluid, stayed in place for a day less than the Portovac units and were emptied once less often.


Hypertrophic scarring: Dexon suture in a randomized trial.

In a randomized prospective trial of three methods of skin suture an increased incidence of hypertrophic scarring in paramedian and inguinal wounds has been found to be associated with the use of 2/0 Dexon suture subcuticularly. However, the variance associated with the type of suture was less than that associated with the sex of the patient. The patient's opinion of the appearance of the scar was the same for all three suture methods used in skin closure.


Recurrent oral ulceration treated with Mysteclin: a controlled study.

Twenty patients with recurrent oral ulceration participated in a placebo-controlled, double-blind trail of Mysteclin syrup (tetracycline hydrochloride and amphotericin) used as a mouthwash. Though a small, consistent improvement occurred with placebo, there was a significant reduction in mean pair scores and numbers of new ulcers recorded daily during the active-treatment periods, the effect lasting for at least four weeks after treatment was stopped. In contrast to topical steroid preparations, Mysteclin syrup is efficacious when begun at any stage of the disorder and is not associated with adverse systemic effects.


Investigation and treatment of amenorrhoea resulting in normal fertility.

A simple scheme of investigation and treatment to restore fertility in amenorrhoeic women is described. Fifty-nine patients with amenorrhoea not due to primary ovarian failure were treated variously as appropriate, mainly with clomiphene (25), bromocriptine (15), or human menopausal gonadotrophins (12), and six by diet to increase their weight. All ovulated, and by the end of the study 55 (93%) had conceived, 42 (71%) had delivered at least one surviving child, and five others (8%) were pregnant and awaiting delivery. Conception rates were 49% within two cycles of treatment and 66% within three cycles; using life-table method to standardise the cumulative conception rates by correcting for patients who did not continue as long as others in the study, the expected conception rate was 79% in six cycles, 94% in 12 cycles, and 98% after 16 cycles. The multiple pregnancy rate was 13% and abortion rate 22%. Delivery rate (for a viable baby) were 48% within 11 months of starting treatment and 53% within one year; expected rates were 76% in 18 months and 97% in two years. The results show that a relatively simple scheme of classifying amenorrhoeic disorders endocrinologically followed by treatment directed at inducing ovulation allows amenorrhoeic women without primary ovarian failure to achieve conception and delivery rates equal to those in normal women.


Neurotensin-containing cell bodies, fibers and nerve terminals in the brain stem of the rat: immunohistochemical mapping.

Neurotensin immunoreactive perikarya, fibers and nerve terminals, visualized by the indirect immunohistofluorescent method in colchicine-pretreated animals, are localized in many discrete regions of the rat brain stem. Cell body groups are found in the inner aspect of the substantia gelatinosa of the caudal trigeminal nuclear complex, the nucleus of the solitary tract, the parabrachial nuclei, the locus coeruleus, the dorsal raphé nucleus, the periaqueductal gray matter, and the ventral tegmental area of Tsai. These areas of cell body density are accompanied by concentrations of fibers and terminals, while the occasional positive perikaryon noted in the dorsal cochlear nucleus is accompanied by only sparse fluorescent fiber/terminal patterns. Other brain stem regions, such as the floor of the fourth ventricle and aspects of the caudal ventrolateral reticular formation, possess substantial numbers of fibers and terminals that are not accompanied by cell bodies. Many aspects of this distribution coincide with the brain stem distribution of the enkephalin pentapeptides, though significant differences in localization are also evident. Interactions of neurotensin with other neurotransmitter candidates are also suggested by its presence in areas enriched in norepinephrine, dopamine, serotonin, and substance P. Certain neurotensin localizations suggest an association of the peptide with functional brain systems preferentially involving these regions. In particular periaqueductal gray and substantia gelatinosa neurotensin synapses are plausible sites for the analgesia elicited after intercisternal injection of low doses of neurotensin.


Multiple isoacceptor forms of several transfer ribonucleic acids in a mutant yeast strain.

By use of reverse phase 5 chromatography, a strain of Saccharomyces cerevisiae (XB 109-5B) has been shown to exhibit multiple isoaccepting forms for several of the transfer ribonucleic acids (tRNAs). This is in contrast with a standard wild-type strain where only one acceptor is found for each tRNA studied. Multiple peaks for tRNATyr, tRNAPhe, tRNASer, and tRNAVal have been detected for strain XB 109-5B. However, the observation of multiple isoacceptors cannot be extended to all tRNAs in this strain since tRNAAsp appears as a single form that is the same as in the wild type. The appearance of multiple peaks was found to depend on the growth conditions of the cells. The tRNA profiles of XB 109-5B that was grown rapidly with vigorous aeration differed the most from profiles of comparably grown wild-type yeast, whereas tRNA from this mutant, grown without shaking or supplementary aeration, appeared the same as the wild type. The minor nucleoside composition of the isoacceptors of tRNAPhe was obtained.


Levels of glutathione in Escherichia coli.

Log phase cells of Escherichia coli growing in minimal medium contain a basal level of glutathione (5 pmol/mL per Klett unit) which can increase more than sixfold when the cells reach stationary phase. Since the addition of cysteine alone to log phase cells illicits the same response, the increase in the intracellular pool of glutathione appears to be influenced by the amount of cysteine available for glutathione synthesis. Glucose depletion at low cell densities resulted in a decrease in the glutathione pool while the addition of amino acids other than cysteine did not affect the glutathione pool. Depletion of ammonia or proline as the nitrogen source also resulted in a decrease in the glutathione pool to one-third of the original basal levels as did a shift to anaerobic growth. The large glutathione pool in stationary phase cells dropped from 31.5 to 4.5 pmol/mL per Klett unit within 30 min of transfer to fresh medium. There was no apparent correlation between changes in the glutathione and coenzyme A--glutathione disulfide (CoASSG) pools after a variety of metabolic disruptions.


Corneal endothelial cell function after storage in MK medium and hydrocortisone.

We stored rabbit corneas either in MK medium or MK medium plus hydrocortisone 10(-6)M for 7 days. The corneas stored in MK medium with hydrocortisone had better temperature reversal characteristics than those stored in MK medium alone.


[Surgical correction of corneal astigmatism after corneal graft].

We performed Troutman's technique of corneal wedge resection in 6 patients who had substantial astigmatism after penetrating corneal grafts. The astigmatism was reduced by 5 to over 15 dioptres, and was stable for up to 36 months. The visual acuity was considerably improved in 5 of the 6 cases. However we could not neither predict the amount of correction nor the axis of the residual astigmatism with this technique. We stress the importance of using a keratometer when performing a penetrating corneal graft.


Immunological aspects of superior limbic keratoconjunctivitis.

Three patients with superior limbic keratoconjunctivitis (SLK) were treated by conjunctival resection. The results of immunological studies and histological examination of the excised tissue were unremarkable; none of seven criteria of antibody dependence were met. We found no specific immune defects in three patients with superior limbic keratoconjunctivitis.


Live influenza vaccine: screening of attenuated virus strains by a 50% ciliary activity inhibition test in organ cultures of ferret trachea.

This study of three live attenuated inhibitor-resistant influenza vaccines showed that these preparations are usually antigenic and that they caused no significant reactions when characterized by an index of attenuation equal to or slightly better than 1.0 arbitrarily attributed to the 'reference' attenuated A/Hong Kong/68 strain of Beare and Bynoe. This index, measured in vitro on ferret tracheal rings, is expressed as the ratio of the time required for ciliary activity inhibition of 50% of the rings by the tested candidate vaccine strain and the 'reference' attenuated strain. Induction of heterologous antibodies was also observed. Oral administration of underattenuated perparations did not cause the severe reactions which were observed when the same vaccine was administered intranasally.


[Regulation of catalase synthesis in Proteus mirabilis].

During the log-phase growth of Proteus mirabilis the specific activity of catalase decreases, while at the beginning of or during the stationary phase an increase takes place which is abolished by inhibitors of nucleic acid or protein synthesis. Glucose in the culture medium has no appreciable effect on the level of enzyme synthesis nor does the passage of bacteria to anaerobiosis bring any noticeable change. Successive additions of hydrogen peroxide up to weak final concentrations (0.2--0.5 mM) stimulate catalase synthesis. Determination of the enzyme in vivo reveals but a weak proportion of the total catalase which can only be titrated after the breakdown of cells. The titrable enzyme in vivo represents, as an order of magnitude, the activity found associated with the cell wall, in an easily released form after the mechanical separation of the inner and outer membranes. Thus, bacteria can act upon exogenous peroxide only through a peripheral catalase while they possess in a masked form an important reserve of cytoplasmic enzyme.


[Properties of an hydrogen peroxide resistant Proteus mirabilis mutant].

A peroxide-resistant mutant (PR) was isolated from Proteus mirabilis using the hydrogen peroxide mutagenic property. Under the same conditions, resistance of mutant PR bacteria to H2O2 was 50 to 100 times greater than that of the wild type. The total amount of catalase produced by P. mirabilis PR was on the average 10 times greater than that of the wild type. When PR bacteria were subjected to high doses of H2O2 (150mM), the determination of catalasic activity in vivo increased; paradoxically, there was a net decrease in the activity of the solubilized catalase after the breakdown of the cells. The hypothesis of an enzyme transfer from the inside towards the periphery of the cells is discussed. The behavior of a membrane enzyme (L-phenylalanine oxidase) of the PR mutant shows that H2O2 may cause lesions way up to the internal membrane of bacteria.


A tentative direct microscopic method for counting living marine bacteria.

Yeast extract (0.025%) and nalidixic acid (0.002%) were added to seawater samples and the samples were incubated for 6 h at 20 degrees C in the dark. Under these conditions, bacterial cells did not divide but grew to form elongated cells that are easily recognized by a direct microscopic method and epifluorescent microscopic technique. The number of cells thus obtained is proposed as a direct cound of viable bacterial cells (DVC). With open ocean samples, DVC was higher than 'viable' plate counts by up to three orders of magnitude and lower than the direct counts by about one order.


A comparative trial of transsphenoidal hypophysectomy and estrogen suppression with aminoglutethimide in advanced breast cancer.

We compared two treatment regimens, transsphenoidal hypophysectomy and estrogen suppression with aminoglutethimide in women with metastatic breast carcinoma. Three of fourteen patients experienced partial objective tumor regression with a median duration of 4.6 months following hypophysectomy, whereas 10 of 21 women receiving aminoglutethimide responded (2 complete, 8 partial) with a median duration of 11.5 months. Side effects in the medical group were minimal while surgical complications included 2 cases of CSF rhinorrhea, one leading to meningitis and death. In patients receiving aminoglutethimide, urinary free cortisol and plasma dehydroepiandrosterone sulfate fell significantly as did plasma estrone and estradiol. In the hypophysectomy group, anterior-pituitary function testing postoperatively revealed adequate suppression of gonadotropin and prolactin secretion but incomplete inhibition of the ACTH-cortisol axis in 4 of 7 surgical patients studied. Five patients initially treated with hypophysectomy experienced a further reduction of plasma (and urinary) estrone and estradiol levels when given aminoglutethimide. We conclude that estrogen suppression therapy with aminoglutethimide is a feasible alternative to surgical hypophysectomy in providing endocrine suppression and palliation in advanced breast carcinoma.


Improved remission rates and remission duration in young women with metastatic breast cancer following combined oophorectomy and chemotherapy: a study of Cancer and Leukemia Groupe B.

One hundred fifty-seven premenopausal women with metastatic breast cancer were prospectively randomized to treatment consisting of oophorectomy + vincristine, prednisone, cyclophosphamide, methotrexate, 5-fluorouracil (VPCMF) (Reg. I) or oophorectomy + cyclophosphamide (Reg. II) or oophorectomy followed by an observation period (Phase 1), followed by VPCMF (Phase 2) (Reg. III). Complete plus partial response rates were 72% on Reg. I, 65% on Reg. II, 18% on Reg. III, Phase 1 and 50% on Reg. III, Phase 2. Median duration of response for Reg. I was 17 months, for Reg. II 16 months, and for Phase 1 and Phase 2 of Reg. III, respectively, 5 months and 8 months. The response rate for patients treated with oophorectomy plus chemotherapy is significantly higher than the response rate achieved with oophorectomy alone as first treatment following the appearance of metastases in premenopausal women.


Cyclophosphamide-induced cardiomyopathy: a report of two cases and review of the English literature.

Fatal cardiomyopathy developed in two patients receiving cyclophosphamide in preparation for bone marrow transplantation. Both patients had normal EKGs prior to receiving cyclophosphamide in total doses of 168 mg/kg (case 1) and 144 mg/kg (case 2) and subsequently developed loss of voltage and ST-T wave changes. One patient (case 1) died of CHF and hypotension while the other patient (case 2) developed tamponade. Prior to this report, the lowest total dose of cyclophosphamide reported to cause fatal cardiomyopathy was 180 mg/kg. In contrast to anthracycline congestive cardiomyopathy, the effects of cyclophosphamide appear to have an acute onset and do not appear to be the cumulative result of drug dosing. Postmortem examination in both patients revealed thickened left ventricles with intramyocardial hemorrhage.


A comparison between combination chemotherapy and total body irradiation plus combination chemotherapy in non-Hodgkin's lymphoma.

Thirty-nine untreated patients with either lymphocytic or nodular mixed/nodular histiocytic non-Hodgkin's lymphoma, stage II--IV, were randomized to treatment with total body irradiation (TBI), 100 rads in 10 fractions over 12 days, plus combination chemotherapy with either cyclophosphamide, vincristine and prednisone (CVP) or cyclophosphamide, vincristine, procarbazine and prednisone (C-MOPP) or to treatment with combination chemotherapy (CVP or C-MOPP) alone. Remission rate and duration was comparable for both treatment groups; thus the use of both treatment modalities ab initio provides no therapeutic advantage.


Anaplastic carcinoma of the thyroid following external irradiation.

Two patients with anaplastic carcinoma of the thyroid following irradiation for Hodgkin's disease are presented, and the previously reported cases briefly reviewed. The risk of late development of thyroid carcinoma is life-long and high-dose irradiation in patients with prolonged life expectancy may result in anaplastic carcinoma as well as less malignant histological forms.


A method for distinguishing benign from malignant breast lesions utilizing antibody present in normal human sera.

A sensitive immunoperoxidase technique was utilized 1) to detect the presence in normal human serum of antibody directed against breast carcinoma cells (tumor associated antibody--TAA) and 2) to determine whether this antibody could be used to discriminate between benign and malignant breast epithelium in sections of formalin fixed paraffin embedded tissue. A total of 16 benign and 13 malignant breast lesions were examined for evidence of binding of human immunoglobulin, before and after application of normal human AB serum containing the putative tumor associated antibody--TAA. Malignant cells showed significant binding of human immunoglobulin (TAA); benign or normal cells did not. Clear immunohistological separation of benign and malignant breast lesions appears possible by this method.


Immunodiagnosis of mesothelioma: use of antimesothelial cell serum in an indirect immunofluorescence assay.

Cells isolated from human serous effusions were cultured in vitro. Monolayers of large multipolar cells were established. Antisera to the cultured cells were prepared in rabbits and rats. The antisera were absorbed with human red cells, liver powder and MOLT-4F cell line lymphocytes. Specificity of the absorbed antisera for human mesothelial cells were demonstrated in an indirect immunofluorescence assay. The antisera were used to confirm the diagnosis of mesothelioma in two cases. In both the patients, the morphologically identifiable malignant cell populations in the effusions stained positively with the antimesothelial cell serum thus establishing their mesothelial origin. Normal nonmesothelial tissue and known nonmesothelial tumors failed to react with the antisera thus confirming the specificity of the antisera.


Immunological skin testing and interpretation: a plea for uniformity.

Immunological skin tests are one of the major methods used for the assessment of one's immune status. A review of the literature clearly reveals that there is no uniform methods for the administration and interpretation of skin tests. Without uniformity, it is obvious that data are not comparable. Contradictory conclusions may be reached from the same skin test data, depending upon the method of interpretation used. Precedent, rather than rationale, has often determined the method of interpretation. There are studies which suggest that slight induration or erythema, often considered negative, are significant immunologic events. The various methods of skin test administration and interpretation are reviewed. Recommendations for application and interpretation are presented so that uniformity may exist.


Cell-surface characteristics of hairy cell leukemia in seven patients.

Surface marker studies were performed on "hairy cells" from 7 patients with hairy cell leukemia (HCL). Using sensitive analytic techniques including specific antisera and Fluorescence Activated Cell Sorter (FACS-1), further definition of the abnormal cell was achieved. Four different antisera were used in infestigating the cell surface characteristics of these patients: anti-p23,30, an antiserum reactive with B cells and a subset of monocytes, anti-311, which reacts only with T cells, pepsin digested anti-F(ab')2 which reacts with B cells only and pepsin digested anti-lysozyme reactive with monocytes and myeloid cells, but not with B or T cells. In all cases strong reactivity was observed with anti-p23,30 and anti-F(ab')2, but no reactivity with anti-311. Five out of the seven cases were reactive with anti-lysozyme in a pattern similar to normal monocytes. Furthermore, when cells were separated according to binding to anti-p23,30, anti-F(ab')2 and anti-lysozyme and in two cases, according to cell size, the majority of reactivity and large cells were "hairy" when examined under microscopy. In contrast, the small and nonreactive (dull cells) appeared as normal mature lymphocytes. Thus, our data supports the view that HCL cells bear in most cases B cell and monocytic membrane markers.


Chromosomes and causation of human cancer and leukemia. XXXII. Unusual features of Ph1-positive acute myeloblastic leukemia (AML), including a review of the literature.

Five cases of Ph1-positive AML were studied. In all cases a Ph1-chromosome was shown with banding techniques to be due to a translocation between chromosomes No. 9 and No. 22. Cases 1 and 4 were found to have more than one Ph1 with evidence of only on Ph1-translocation accompanying other chromosome abnormalities. Two cases represented an unusual pattern of appearance and disappearance of the Ph1-positive clone during their clinical courses: Case No. 2 was originally Ph1-positive (46,XY,Ph1) but two months before his expiration the Ph1-positive clone was completely replaced by a newly developed Ph1-negative clone with an abnormal chromosome No. 21 (46,XY,21q+), whereas case No. 3, primarily Ph1-negative, developed a Ph1-positive clone among the previously karyotypically normal cells one month before death. The Ph1-positive AML cases presented have been discussed in relation to: 1) the genesis and significance of the Ph1-positive clone, 2) differentiation from the blastic phase of CML and 31 the general experience with Ph1-positive acute non-lymphocytic leukemia (ANLL), the world literature of which have been tabulated.


Steroid localization in granulosa-theca tumors of the ovary.

Eleven granulosa-theca tumors (seven pure granulosa and four with associated theca elements) were examined for the presence of estradiol, progesterone and testosterone using an indirect immunoperoxidase technique on formalin-fixed paraffin embedded tissue. In ten patients the endometrium was studied histologically and significant endometrial hyperplasia consistent with estrogen production by the tumors was found. Estradiol was localized in granulosa cells (nonluteinized) in all 11 cases and in luteinized theca cells in three of the four cases in which theca elements are present. In contrast, progesterone was always detected in luteinized theca cells and in granulosa cells in over one half the cases. Testosterone was also present in granulosa cells in just over half the cases but tended to be only weakly positive. The nonluteinized stromal cells were negative for all steroids. These results are compatible with the concept that in granulosa-theca tumors, both granulosa and theca cells can produce a wide range of steroid hormones but that the predominant steroid present in granulosa cells is estradiol, while progesterone is the predominant steroid in luteinized theca cells.


Adenocarcinoma of the pancreas associated with hypoglycemia: case report and review of the literature.

The occurrence of profound hypoglycemia in a patient with metastatic adenocarcinoma of the pancreas is reported. In contrast to the four previously reported cases, no suggestion of excess insulin production was found. Metabolic studies in this patient suggest both increased peripheral glucose utilization and decreased hepatic glucose production as contributing factors which promoted the hypoglycemia.


Differences in age and sex distributions among patients with non-Hodgkin's lymphoma.

The records of a group of 337 adult patients with non-Hodgkin's lymphoma seen at the Stanford University Medical Center, Division of Oncology were examined for relationships between stage and histopathological classification and simple demographic characteristics. Patients with Stages I and II of disease and diffuse varieties of lymphoma were found to be younger than patients in other categories. An excess of male patients was noted particularly in younger patients with diffuse lymphoma and Stages I and II of disease. Male patients with Stages I and II disease were noted to be bimodally distributed with respect to age, with peak number of patients in the fourth and sixth decades. This was particularly apparent among patients with diffuse histiocytic lymphoma. The implications of these findings are discussed.


Ultrastructural and immunohistological study of immunoblastic sarcoma developing in child with immunoblastic lymphadenopathy.

The first case of immunoblastic lymphadenopathy developing in a child which progressed to immunoblastic sarcoma is reported. The sarcoma cells showed light and electron microscopic features of transformed lymphocytes (immunoblasts) but it was not possible to establish their B-cell origin using a peroxidase-antiperoxidase technique for the demonstration of intracellular immunoglobulins. In the affected lymph nodes there was marked proliferation of reticulum and endothelial cells both of which contained numerous intranuclear inclusions which may be of viral origin. Ultrastructural studies suggest that the amorphous eosinophilic interstitial material, an important diagnostic morphological feature of immunoblastic lymphadenopahy, results from the oblique sectioning of the elongated and branching cytoplasmic processes of reticulum cells. It is postulated that in immunoblastic lymphadenopathy the proliferation of reticulum and endothelial cells may be the primary event, perhaps stimulated by viral infection and that the intense lymphocytic and plasmacytic infiltration and sarcomatous transformation occur as secondary phenomena.


Childhood lymphoma in southern Iran.

A study of 81 childhood lymphomas diagnosed in the Department of Pathology of Pahlavi University Medical Center, Shiraz, Iran, encompassing all histologically diagnosed childhood lymphomas from the Fars Province, Southern Iran over a 14-year period (1963--1976) revealed a 3:1 male predominance and a 1:4 frequency compared to adult lymphomas. Peripheral lymphadenopathy at the initial physical examination was almost twice as common as deep node involvement. Comparison of cumulative and age-standardized (to world population) incidence rates with those of selected Tumor Registries in various continents revealed a higher rate in our region of both non-Hodgkin's and Hodgkin's lymphoma relative to some of the Western countries. Our incidence rates were in general intermediate between Western populations on one hand and some South America, African and Asian populations on the other. Hodgkin's disease accounted for 64% (males) and 88% (females) of lymphomas and mixed cellularity was the commonest histologic subtype. Histologically almost all non-Hodgkin's lymphomas were diffuse at the time of diagnosis.


Unusual manifestations of non-Hodgkin's lymphoma.

Two patients with biopsy proven lymphomatous infiltration of uncommon sites are presented. One had histiocytic lymphoma, involving the true vocal cord with a squamous cell carcinoma on the opposite vocal cord. The other had diffuse well-differentiated lymphocytic lymphoma, presenting with nodal and periorbital disease. After local radiation therapy and while clinically free of other lesions, he developed hemotochezia due to several polypoid lesions throughout the entire colon and prostatic infiltration with symptoms of lower urinary tract obstruction. Both problems were solved after specific treatment.


Metastasizing extradural ependymoma of the sacrococcygeal region: case report and review of literature.

A case is discussed in which the patient presented with a primary extradural sacrococcygeal ependymoma and synchronous pulmonary metastasis. The clinical course has been characterized by recurrent pulmonary metastases. Management has consisted of repeated surgical resections of the pulmonary metastases and the tumor at the primary site; and the use of a wide spectrum of chemotherapeutic agents. Transplantation of this tumor into nude mice initially resulted in rapid growth but there was spontaneous regression in the second transplants. A general discussion of the management of such lesions is presented, and the literature pertaining to this tumor is discussed.


Phase II evaluation of ftorafur in previously untreated colorectal cancer: a Southwest Oncology Group Study.

Eighty-four previously untreated patients with metastatic adenocarcinoma of the large intestine received intravenous ftorafur at a dosage of 2.25 g/m2/day for 5 consecutive days. Courses were repeated every three weeks. Regressions were noted in 9 of 84 treated patients (11%). Median survival for all patients was 32 weeks. Responders survived only 5 weeks longer than nonresponders; 36 vs. 31 weeks. Central nervous system toxicity was a limiting factor occurring in one-third of patients. Ftorafur in a daily X5 schedule appears not to make a significant contribution to the management of disseminated colorectal cancer.


Mutagenic activity of marihuana smoke condensates.

Smoke condensates prepared from marihuana cigaretts were mutagenic in strain TA98 of the Ames Salmonella/microsome test, a short-term bioassay which estimates the mutagenic and carcinogenic potential of some chemicals. The mutagens in marihuana smoke condensates required liver enzymes to be activated. The specific mutagenic activity of marihuana smoke condensates were similar to that of tobacco smoke condensates prepared from American cigarettes. Fractionation studies of the marihuana smoke condensates showed that basic components accounted for 76% of the recovered mutagenic activity.


A nucleolar antigen found in a broad range of human malignant tumor specimens.

With rabbit antibodies to nuclear 0.01 M Tris-HCl, pH 8, extract or "nucleolar preparations" of human HeLa S3 cells and fluorescein-labeled goat anti-rabbit antibodies, bright nucleolar immunofluorescence was observed in 61 or 63 human adenocarcinomas, squamous cell carcinomas, sarcomas, hematological neoplasms, and other malignant tumors. With these antibodies, nucleolar immunofluorescence was not found in 23 normal tissue specimens, 10 benign adenomas and hyperplastic tissues, and 8 specimens of inflammatory diseases. In the nontumorous tissues examined, positive nucelolar fluorescence was found in a few sections of a gastric ulcer and chronic ulcerative colitis which have been known propensities for malignant change; these areas may have been undergoing focal malignant changes.


Human urinary metabolites of 1-(tetrahydro-2-furanyl)-5-fluorouracil (ftoraful).

Two hydroxylated metabolites were isolated from the urine of a patient who had received ftorafur (5 g/sq m). These metabolites were identified by mass spectrometry and nuclear magnetic resonance spectroscopy as trans-3'- and cis-4'-hydroxyftorafur. The compounds were not converted to 4-fluorouracil when incubated in plasma, base (pH 9), or water. Because of their stability, it is unlikely that these metabolites are in vivo precursors of 5-fluorouracil. There are indications that less stable, unisolatable, hydroxylated ftorafur derivatives are intermediates in the conversion of ftorafur to 5-fluorouracil.


The distribution of vasopressin-, oxytocin-, and neurophysin-producing neurons in the guinea pig brain. I. The classical hypothalamo-neurophypophyseal system.

The location, cytology and projections of vasopressin-, oxytocin-, and neurophysin-producing neurons in the guinea pig were investigated using specific antisera against vasopressin, oxytocin or neurophysin in the unlabeled antibody enzyme immunoperoxidase method. Light microscopic examination of the neurons of the supraoptic and paraventricular nuclei shows that hormone is transported not only in axons, but also in processes having the characteristics of dendrites. Neurons were found to contain only vasopressin or oxytocin; all neurons containing neurophysin appear to contain either vasopressin or oxytocin. In the neural lobe, vasopressin and oxytocin terminals are intermingled. In the median eminence, vasopressin and oxytocin fibers are intermingled in the internal zone. In a caudal portion of the median eminence, a number of vasopressin and neurophysin (but few oxytocin) axons enter the external zone from the internal zone, and surround portal capillaries. In the supraotic nucleus, vasopressin neurons outnumber oxytocin neurons with a ratio of at least 5:1. The paraventricular nucleus is separated into two distinct groups of neurons, a lateral group consisting of only vasopressin neurons, and a medial group consisting of only oxytocin neurons. In addition to axons passing to the neurohypophysis, a number of axons appear to interconnect the supraoptic and paraventricular nuclei.


Ontogenesis of cells producing polypeptide hormones (ACTH, MSH, LPH, GH, prolactin) in the fetal hypophysis of the rat: influence of the hypothalamus.

The ontogenesis of cells containing polypeptide hormones (ACTH, MSH, LPH, GH and Prolactin) was investigated in the fetal rat hypophysis by immunohistochemistry using the peroxidase-antiperoxidase complex. Corticotrophs, melanotrophs and lipotropic cells were revealed earlier in the pars distalis than in the pars intermedia. In the pars distalis, cells producing LPH were found in the morning of day 15 of gestation using anti-gamma- or anti-beta-LPH sera, and in afternoon using anti-alpha- or beta-endorphin sera. Cells containing beta-MSH were observed from the afternoon of day 15. The cells stainable with the anti-alpha-MSH, anti-beta-(17--39)ACTH and anti-beta-(1--24)ACTH sera appeared on day 16. In the pars intermedia, the cells producing alpha-MSH, beta-MSH, alpha- and beta-endorphin, gamma- and beta-LPH were observed in the morning of day 17, while cells containing ACTH were only revealed in the afternoon of the same day of gestation. Based on the treatment of serial paraffin sections with various antisera, it was clearly shown that MSH, ACTH, and LPH occur in the same cells located in the pars distalis as in the pars intermedia. The development of the corticotrophs, melanotrophs and lipotropic cells does not require the presence of the fetal hypothalamus or other central nervous structures. The pituitary glands of 21 day-old fetus encephalectomized on day 16 showed as many reactive cells as those of the littermate controls. The somatotrophs were first revealed in the pars distalis in the afternoon of day 19. The cells producing prolactin were not observed before day 21 of gestation. On some cases GH and prolactin were found together in one cell. The cytodifferentiation of GH and prolactin cells is apparently not under hypothalamic control.


Immunocytochemical localization of a calcium-binding protein in the rat duodenum.

The cellular localization of the vitamin D-dependent calcium-binding protein (CaBP) in the duodenum of rat was studied using indirect immunofluorescence and immunoperoxidase-staining methods. Specific positive reaction product, indicative of the presence of CaBP, was exclusively located within the villous part of the duodenal mucosa. Moreover, CaBP was detected mainly within the supranuclear region of the cytoplasm of absorptive cells and also at the level of their basal laminae. CaBP was not demonstrable either in the nuclei or associated with the brush border membrane of absorptive cells. Also, CaBP was neither detectable in goblet cells nor in sub-epithelial layers. When the specific anti-CaBP antiserum was replaced by nonimmune rabbit serum or when it was preabsorbed on a CaBP-Sepharose conjugate, no positive immunostaining was seen. Together with recent biochemical data our observations agree well with the view that CaBP may act as an intracellular "buffer" by protecting the cell against too high Ca2+ concentrations.


Indication for a granule-free form of vasopressin in immobilization-stressed rats.

Following immobilization stress the supraoptic nucleus exhibits an increased number of coarse, heavily immunostained fibers in the basal glia labyrinth. Ultrastructural immunocytochemistry demonstrates a labeling of the rough endoplasmic reticulum in the neurosecretory perikarya and granule-free, immunoreactive material in the axons adjacent to the basal glia labyrinth. Furthermore, a labeling of the intercellular clefts of the neuropil is demonstrable in the supraoptic nucleus. These results lead to the hypothesis that 1) vasopressin is synthesized and released in two forms, in a granule-bound form and in a granule-free, probably more soluble form, and that 2) the latter might be released already in the nuclear area into the intercellular clefts from where it may reach its target cells via the cerebrospinal fluid of the subarachnoid space.


Immunocytochemical localisation of prolactin and growth hormone in the perinatal sheep pituitary: a morphological and quantitative study.

The proxidase anti-peroxidase immunocytochemical staining technique has been used to identify prolactin and growth hormone cells in pituitaries from fetal and neonatal sheep. The size of the secretory granules in these cell types has been measured using the image analysing computer Quantimet 720. The area size distributions of the fetal prolactin and growth hormone granules were compared with those in the neonate and the adult. It appears that the gestional age of the fetus may influence the size range of prolactin secretory granules.


A secondary attachment site for bacteriophage lambda in trpC of E. coli.

We have determined the nucleotide sequence of a secondary lambda attachment site in trpC. Direct sequence analysis of lambdatrp transducing phage DNA fragments carrying the two prophage attachment sites reveals a 6 nucleotide homology in the crossover region which is a subset of the 15 nucleotide core sequence in the primary lambda attachment site: GCTTTTTTATACTAA. This 6 nucleotide sequence is also present in the intact trpC genome at the attachment site, as shown by analysis of trpC mRNA spanning this region.


Isolation of galactose-inducible DNA sequences from Saccharomyces cerevisiae by differential plaque filter hybridization.

Multiple nitrocellulose DNA filter replicas of plaques of in vitro generated recombinants of phage lambda and Saccharomyces cerevisiae have been screened by hybridization with 32P-labeled cDNA probes. These probes were representative of total poly(A)-containing RNA of yeast cells grown on acetate, galactose, glucose or maltose. This approach allows the use of specific differences in total RNA populations as probes for gene isolation. Five "galactose-induced" clones have been isolated. Expression of the RNA coding regions on at least two cloned sequences, Sc481 and Sc482, is regulated by genes known to control the expression of the structural genes required for the conversion of exogenous galactose to endogenous glucose-1-phosphate. One cloned sequence, Sc484, is expressed during growth on all carbon sources except glucose, and is not under control by the galactose regulatory genes. This clone contains a sequence that is repeated 3 times in the yeast genome. The cloned fragment Sc481 contains coding regions for all or part of three galactose"induced RNAs and may correspond to the GAL 1, GAL 7, GAL 10 gene cluster region of chromosome II.


Structure and organization of the two tRNATyr gene clusters on the E. coli chromosome.

The structure and organization of the gene clusters coding for the two tyrosine-accepting tRNA species (tRNA1Tyr and tRNA2Tyr) on the E. coli chromosome have been determined. The mature structural sequences of the two tRNATyr genes, located on opposite sides of the E. coli chromosome, differ by only 2 bp, but sequences surrounding these portions of the genes are very different. The genes coding for tRNA1Tyr (tyrT) comprise two mature structural sequences separated by a 200 bp "intergenic spacer." It is known that in transducing phage, the region adjoining the CCA end of the second mature structural sequence comprises a 178 bp repeated sequence which contains an in vitro, rho-dependent transcriptional termination site. We find that these potentially genetically unstable repeated sequences are present in the E. coli chromosome with the same organization as that determined from transducing phage analyses. The gene that codes for tRNA2Tyr (tyrU) is present in a single copy and is tightly clustered with three other tRNA genes. One of these genes (to be called thrU) encodes a previously undescribed tRNA (to be called tRNA4Thr). The organization of this cluster on the E. coli chromosome is tRNA4Thr--8 bp--tRNA2Tyr--115 bp--tRNA2Gly--6 bp--tRNA3Thr. The importance of correlating structural analyses derived from specialized transducing phage with those determined for the chromosome itself is demonstrated by results which show that out of four independently isolated tRNATyr transducing phage, two carrying the tRNA1Tyr genes [phi80psu3+,- (Cambridge) and phi80sus2psu3+ (Kyoto)] and two carrying the tRNA2Tyr gene (lambdarifd 18 and lambdah80dglyTsu+36), only the first phage from each group has the same gene organization as that found in the E. coli chromosome.


Mutants in transmission of chemotactic signals from two independent receptors of E. coli.

We have characterized chemotactic mutants of E. coli that appear to be defective in a common linkage of two independent receptors to the central chemotactic components. The mutants do not respond to gradients of ribose or galactose and thus are called trg (taxis to ribose and galactose), after Ordal and Adler (1974b). These trg mutants are indistinguishable from their parent in tactic response to other attractants, swimming pattern, growth rates, and transport of ribose and galactose. The mutant cells contain the usual amounts of ribose and galactose receptors, and those proteins function normally in their other role, transport of their respective ligands. The mutations, generated by insertion of translocatable drug-resistance elements (transposons)8 are located near 31 min on the map of the E. coli chromosome, a locus far removed from the genes coding for the ribose and galactose receptors. Trg mutants do not resemble either specific receptor mutants or che mutants. The nature of the requirement for the trg product in the response to ribose and galactose is not defined, but evidence for interference of tactic signals from the ribose and galactose receptors (Strange and Koshland, 1976) supports the idea that the product functions directly in the transmission of tactic signals from the two receptors to the flagella.


DNA sequence analysis of Tn10 insertions: origin and role of 9 bp flanking repetitions during Tn10 translocation.

The sequences of insertions of the translocatable tetracycline-resistance element Tn10 into the repressor (cl) gene of bacteriophage lambda have been analyzed. Each insertion contains the same discrete set of Tn10 sequences flanked by a direct repetition of a 9 bp cl-gene sequence. The flanking repititions are generated by duplication of information present only in the target DNA molecule rather than by a Campbell-type recombination event between one 9 bp sequence on the target DNA and a second one provided on the incoming element. The repetitions do not contain genetic or structural information important for translocation. A genetically constructed Tn10 insertion which lacks flanking repetitions is fully functional in translocation to a new position. Tn10 insertions cluster at preferred positions along a target DNA (Kleckner et al., 1979). Sequence analysis shows that four independently isolated cl::Tn10 insertions occur at identical positions in the cl gene. We speculate that homology between Tn10 and its target, at some distance from the site of the actual recombination event, could be relevant to the preference of Tn10 for particular insertion sites.


Evidence for transposition of dispersed repetitive DNA families in yeast.

Dispersed repetitive DNA sequences from yeast (Saccharomyces cerevisiae) nuclear DNA have been isolated as molecular hybrids in lambdagt. Related S. cerevisiae strains show marked alterations in the size of the restriction fragments containing these repetitive DNAs. "Ty1" is one such family of repeated sequences in yeast and consists of a 5.6 kilobase (kb) sequence including a noninverted 0.25 kb sequence of another repetitious family, "delta", on each end. There are about 35 copies of Ty1 and at least 100 copies of delta (not always associated with Ty1) in the haploid genome. A few Ty1 elements are tandem and/or circular, but most are disperse and show (along with delta) some sequence divergence between repeat units. Sequence alterations involving Ty1 elements have been found during the continual propagation of a single yeast clone over the course of a month. One region with a large number of delta sequences (SUP4) also shows a high frequency of sequence alterations when different strains are compared. One of the differences between two such strains involves the presence or absence of a Ty1 element. The novel joint is at one inverted pair of delta sequences.