Person-to-person spread of Salmonella typhimurium after a hospital common-source outbreak. In September, 1973, diarrhoea caused by Salmonella typhimurium developed in 32 people in a Maine hospital.  Both epidemiological and microbiological evidence indicated that raw egg beaten in milk ("egg-nog") was responsible for the infection.  However, 6 patients and 8 employees had not had egg-nog, and their illness developed after the source of infection had been recognised and removed.  Most of these people had had direct contact with an infected patient, and presumably acquired the infection by person-to-person spread.  It is concluded that person-to-person spread of S. typhimurium can occur in hospitals and can be a hazard to patients and staff.


Pollution, noise, and mental health.

Pollution, noise, and mental health. Though the adverse psychological effects of noise as an environmental pollutant are well recognised, much of the relevant work has been focused on the ambiguous concept of "annoyance". The relation between noise and mental ill-health calls for more direct investigation. In particular, the pronounced individual variation in reactions to noise requires elucidation.


Granulocyte transfusions in treatment of infections in patients with acute leukaemia and aplastic anaemia. By use of the continuous-flow blood-cell separator 137 bags of granulocyte-rich plasma were obtained from normal donors (59 bags) and patients with chronic granulocytic leukaemia (C.G.L.) (78 bags). Eighty-nine courses of granulocyte transfusion therapy consisting of 1 or more such bags were administered to forty-one ABO-compatible patients with acute leukaemia or aplastic anaemia, who had definite or probable infections that had failed to respond to antibiotics. The fever resolved after 67% of courses of transfusions of two or more bags but after only 24% of transfusions of single bags of granulocytes (p less than 0-01), and this result suggests that this form of treatment is in general effective. Granulocytes from C.G.L. and normal donors were equally effective, although transfusion reactions were commoner after C.G.L. cells (33% versus 12%, respectively, p less than 0-05). C.G.L. grafts, and probable graft-versus-host disease, occurred in three recipients of unirradiated C.G.L. cells. Recipients of normal cells whose fevers resolved received on average four times as many granulocytes per sq.m. as those fevers did not respond. No such difference was found when C.G.L. cells were used. The fever was more likely to resolve in recipients with established or clinically probable bacterial or fungal infections than in those with fever of uncertain cause. Fever was less likely to resolve in recipients with peripheral blood granulocyte counts before transfusion of greater than 1000 per mul. It is concluded that granulocyte transfusion therapy is a valuable advance in the management of infections in neutropenic patients.


Controlled trial of therapy in covert bacteriuria of childhood. Sixty-three girls with covert bacteriuria were included in a controlled trial of therapy. Recurrent infection in the treated group was common and was not significantly different from the rate of persistent infection in the untreated control group. Two children in each group developed clinical pyelonephritis; the others have remained healthy and all of them have a normal rate of growth. 2 years after diagnosis three of the thirty-four children in the control group and one of twenty-six children in the treated group have radiological evidence of new scars of pyelonephritis. These changes were relatively minor and in both groups of children renal growth was similar to that in normal children. It is suggested that for most of these children therapy is not essential, and that when renal changes occur they are of little or no significance. Prescriptive screening for cobert bacteriuria of childhood cannot be recommended at present.


Thyrotoxicosis due to "silent" thyroiditis. 3 patients (2 male, 1 female) presented with symptoms of thyrotoxicosis associated with elevated blood-levels of thyroid hormone and a markedly depressed thyroidal uptake of 131-I. The male patients (aged 59 and 47) each had a cardiac arrhythmia, but did not have any thyroid pain or swelling. The female with a goitre had no discomfort in the neck. Thyrotoxicosis factitia was excluded by history. The subsequent course of their disease was typical of subacute thyroiditis. The elevated thyroid-hormone levels spontaneously fell to normal over a few weeks. In 1 patient, however, chemical hypothyroidism developed. These patients could have been diagnosed as having hyperthyroidism, rather than subacute thyroiditis, since thyroid pain--and swelling in 2 of the cases--was absent. The correct diagnosis was suspected only after finding a thyroidal uptake of 131-I near zero. The thyroidal uptake of 131-I is still important as a routine diagnostic aid in thyroid disease.


Uracil-specific anti-R.N.A. antibodies in scleroderma. Antibodies to single-stranded R.N.A. were found by counter immunoelectrophoresis in all of 40 sera from patients with scleroderma. These antibodies were specific to the uracil bases of R.N.A. Antibodies to R.N.A. were also found in 20 of 40 sera from patients with systemic lupus erythematosus (S.L.E.), but in none of forty controls. Antibodies to R.N.A. found in S.L.E. sera could be differentiated immunochemically from those found in scleroderma in that they were more heterogeneous and could react selectively with either uridine or uridine monophosphate. Antibodies ot D.N.A. were more frequent in S.L.E. than in scleroderma. That antibodies to D.N.A. are actually present in scleroderma and precipitin lines are not the result of cross reactivity with anti-R.N.A. antibodies is indicated by the finding that 10 of the 18 scleroderma sera which reacted with D.N.A. also reacted with thymidine, a base present in D.N.A. but not in R.N.A.


Diogenes syndrome. A clinical study of gross neglect in old age. A study of elderly patients (fourteen men, sixteen women) who were admitted to hospital with acute illness and extreme self-neglect revealed common features which might be called Diogenes syndrome. All had dirty, untidy homes and a filthy personal appearance about which they showed no shame. Hoarding of rubbish (syllogomania) was sometimes seen. All except two lived alone, but poverty and poor housing standards were not a serious problem. All were known to the social-services departments and a third had persistently refused offers of help. An acute presentation with falls or collapse was common, and several physical diagnoses could be made. Multiple deficiency states were found--including iron, folate, vitamin B12, vitamin C, calcium and vitamin D, serum proteins and albumin, water, and potassium. The mortality, especially for women, was high (46%); most of the survivors responded well and were discharged. Half showed no evidence of psychiatric disorder and possessed higher than average intelligence. Many had led successful professional and business lives, with good family backgrounds and upbringing. Personality characteristics showed them to tend to be aloff, suspicious, emotionally labile, aggressive, group-dependent, and reality-distorting individuals. It is suggested that this syndrome may be a reaction late in life to stress in a certain type of personality.


Abnormal organic acidurias in mentally retarded patients. Urine specimens from 1778 mentally retarded patients and 420 age and sex matched non-retarded controls selected from a general practice have been analysed for non-amino organic acids by a quantitative extraction and gas chromatographic method. The compounds were identified by combined gas chromatography and mass spectrometry. Approximately 5% of the patients had an abnormal organic aciduria. The frequency of abnormalities was slightly higher (about 7%) in a group of 248 severely subnormal children, but not in cases with a family history of mental retardation, retarded sibs, or whose parents were consanguineous. The most frequently observed abnormalities were phenylalanine metabolites in cases of phenylketonuria (about 1%), increased excretion of benzoic acid (about 1%), and increased excretion of 2-oxoglutaric acid with or without raised urinary citric-acid levels (about 1%). The biochemical and clinical significance of these findings is being further investigated.


Reversible renal concentrating defect in shock. The loss of renal concentrating power in haemorrhagic shock is reversible upon correction of the shock state. Shock resulting from acute hypovolaemia leads to the following sequence of events: (1) diminished renal blood-flow; (2) decreased superficial cortical nephron perfusion; (3) continued juxtamedullary nephron perfusion; (4) enhanced proximal reabsorption of Na, Cl, and H23; (5) decreased delivery of these ions to the ascending limb, which results in diminished hypertonicity of the medullary interstitium. This hypotonicity is worsened by the "washout" effect on the interstitial hypertonicity caused by continued perfusion of the juxtamedullary vasa recta which results in (6) diminished renal concentrating capacity due to elimination of medullary hypertonicity. Replenishing blood-loss and correcting the hypovolaemic state "regenerates" the hypertonic renal medullary interstitium by (a) diminishing proximal reabsorption and allowing presentation of greater quantities of Na and Cl to the ascending-limb "pump", (b) restoring superficial cortical nephron perfusion, thereby decreasing juxtamedullary perfusion and in this manner eliminating medullary "washout".


Influenza vaccination and mortality from bronchopneumonia in the elderly. In a three-year influenza vaccination programme carried out among elderly patients these were found to have a lower haemagglutination-inhibiting antibody level and a poorer serological response to vaccination than younger persons in the same city. Although there was little difference in overall respiratory illness between the vaccinated and unvaccinated groups until the third year of observation, those who received vaccine showed a substantially smaller incidence of bronchopneumonia and a significantly lower mortality than those not so protected. The observations are believed to justify the giving of influenza vaccine in this age-group.


Tetanus in Haiti.

Tetanus in Haiti. In 1958-72, 985 cases of tetanus (excluding tetanus of the newborn) were admitted to a hospital in Haiti. Mortality was 22%, and in later years (1966-72) mortality fell. During this period the dosage of tetanus antitoxin was lowered to 10,000 units and, for sedation, diazepam has satisfactorily replaced multiple-drug regimens used in earlier years. However, good nursing, including close attention to breathing, is probably the most important item in treatment. As a result of a programme of maternal immunisation, admissions for neonatal tetanus have fallen, and mortality for this condition has been reduced to 26%.


Alveolar rhabdomyosarcoma of the ethmoid sinus. A case of the alveolar rhabdomyosarcoma originating in the left ethmoid sinus was reported. Despite treatment by radical excision of the tumor, irradiation and chemotherapy, the patient died of generalized metastases of the tumor six months after surgery; however, no local recurrence of the tumor was found at an autopsy. The tumor was further studied by light and electron microscopic procedures.


[Gammaglobulin prophylaxis of infectious hepatitis (author's transl)]. When travelling in tropical areas and developing countries with deficient hygienic conditions, a distinctly greater risk of disease is observed with regard to infectious hepatitis. A prophylactic injection of gammaglobulin (5 ml i.m. for adults) for protection against diseases was found to be effective and expedient. A satisfactory protection can be assumed for a period of up to 4 months; after that, the prophylactic injection of gammaglobulin must be repeated in the same dosage, if there is a further risk of infection. It is strongly recommended that the prophylaxis be carried out at least for the first year of exposure to the risk, while later a gradual immunity possibly develops. Children should receive a reduced dose according to age.


The incidence of cerebral amyloid angiopathy in Alzheimer's disease. The incidence and forms of cerebral amyloid angiopathy were studied in 15 cases of Alzheimer's disease using Congo red staining and polarization. Thirteen cases showed slight to severe involvement; two contained no amyloid vascular degeneration. There was a correlation between the presence of amyloid-rich plaques and cerebral amyloid angiopathy (especially the plaque-like angiopathy) but no correlation with "amyloid-poor" senile plaques or Alzheimer's neurofibrillary degeneration.


[Current trends in radiation therapy].

[Current trends in radiation therapy]. The discovery that Hodgkin's disease has a unicentric onset has enabled its prognosis to be modified. In its initial, uniregional stage, in fact, it can be cured by radiotherapy. A satisfactory result will only be obtained, however, if certain fundamental rules are followed: aggressive treatment with extensive radiation fields, radiation of adjacent lymphatic districts, prior splenectomy where necessary.


Nitroblue tetrazolium reduction test in pregnancy. The use of the nitroblue tetrozolium (NRT) dye reduction test as a screening method for bacterial and fungal infections is now widely recognized. Levels have been reported to be falsely elevated in the pregnant state. This report includes a prospective study which showed that the results of this test in 124 pregnant women were indistinguishable from those of a normal control population. Seven of 8 elevated levels were attributable to infection or allergy. With this data, it is concluded that the NBT test can be used to screen for bacterial and fungal illnesses in a febrile pregnant patient as well as in the general population.


Partial amino acid sequence of rabbit beta2-microglobulin. The amino acid sequence of the first 35 residues of a low-molecular-weight protein obtained from the urine of rabbits treated with sodium dichromate was determined and shown to be identical with human beta2-microglobulin in 30 positions. Rabbit beta2-microglobulin, like the human protein, is strikingly homologous to the constant regions of rabbit immunoglobulin G, particularly the CH3 region.


Batrachotoxin block of fast axoplasmic transport in mammalian nerve fibers. Batrachotoxin (BTX) irreversibly blocks fast axoplasmic transport in nerve in concentrations as low as 0.2 micromolar. The action of BTX was studied in cat sciatic nerves in vitro by measuring the rate of the crest outflow after injection of the L7 dorsal root ganglion with [3-H]leucine. Tetrodotoxin, which in itself does not affect fast axoplasmic transport, inhibited the blocking action of BTX. Unlike the BTX block of nerve and muscle membrane excitability brought about through increased permeability to sodium ion, the BTX block of fast axoplasmic transport occurs with or without sodium ion in the medium. High concentrations of calcium ion protected against the blocking action of BTX, while magnesium ion did not. An action of BTX on the transport mechanism inside the fibers was indicated by the small reduction of adenosine triphosphate plus creatine phosphate, which in itself did not account for the block of axoplasmic transport.


Elevated salivary and synovial fluid beta2-microglobulin in Sjogren's syndrome and rheumatoid arthritis. Beta2-Microglobulin is normally present in low concentrations in serum and other bodily fluids. By use of a radioimmunoassay, elevated concentrations of beta2--microglobulin were found in saliva and synovial fluid from patients with Sjogren's syndrome and rheumatoid arthritis, autoimmune inflammatory diseases that attack and destroy the salivary glands and articular tissues, respectively. Elevated beta2-microglobulin concentrations decreased in the saliva of two patients who simultaneously showed a clinical response to systemic treatment. Measurement of beta2-microglobulin in inflammatory fluids may offer a simple method of quantifying local activity in autoimmune states.


Chemotherapeutic drugs increase killing of tumor cells by antibody and complement. When the ascitic forms of two antigenically distinct guinea pig hepatomas induced by diethylnitrosamine are treated in vitro with chemotherapeutic drugs, their sensitivity to killing by xenogeneic antibody plus guinea pig complement increases. The effect is dependent on drug dose, is reversible, and does not appear to be due to increased antigen expression or fixation of the early acting components of guinea pig complement.


Immunohistochemical study of alpha-fetoprotein and serum albumin in the early postnatal period in mice. The localization of the embryo-specific protein alpha-fetoprotein (alpha-FP) and serum albumin in the organs of mice in the first three weeks of life was studied. Using a reaction of immunofluorescence, alpha-FP was detected in the cytoplasm of the hepatocytes. During the first three days of life it was contained in the overwhelming majority of the hepatocytes, but in different concentrations. Then the fluorescence intensity and number of cells containing alpha-FP decreased sharply, and by the seventh to 12th day such cells remained chiefly close to the central veins. In a simultaneous determination of alpha-FP and albumin in the liver, cells containing only alpha-FP, alpha-FP and albumin, and only albumin were detected. With increasing age of the animals, the number of the latter cells increased. On slices of other organs, alpha-FP, albumin, and transferrin were present in the same structures of gamma-globulin.


Prenatal diagnosis of severe central nervous system defect. Report of a successful case. A case is presented in which the latest methods of antenatal diagnosis of central nervous system defect were successfully employed. The advantages and drawbacks of the technique are discussed.


Simultaneous demonstration of lipids and starch in plant tissues. A rapid and easy technique for the simultaneous demonstration of lipids and starch in the same histological section is described. Tissues are prepared by the classical fixing and Araldite M embedding techniques of electron microscopy. Semithin sections are directly stained for 1 hour at 60C with saturated Sudan black B in 70% ethanol without removing the embedding resin. Lipids stain black; starch is shown as white grains contrasting with the blue-grey embedding resin.


Use of peroxidatic-enzyme staining to enhance resolution of cultured mammalian cells under phase microscopy. Staining of glutaraldehyde-fixed mammalian cells with peroxidatic enzymes (horseradish peroxidase or horse heart cytochrome c) greatly enhances resolution of their structure under phase microscopy. The topography of cell processes and regions of intercellular contact and overlapping is resolved precisely, even in dense cultures mounted in media which ordinarily do not permit clear demonstration of these areas. The technique is therefore a useful aid to the study of cultured cells with phase optics. Labeling depends on introducing free aldehydes into cells through the use of bifunctional fixatives such as glutaraldehyde. Acetone or formaldehyde fixation prevents staining, and labeling intensity is greatly diminished by pretreatment with spermine, a polyamine that reacts with glutaraldehyde. Electron microscopy reveals that peroxidase tags membranes preferentially; some areas are labeled smoothly, others in a punctate manner. Ribosomes are sharply contrasted, but nuclei remain unstained. Cytochrome c labels condensed nuclear chromatin intensely, and also stains ribosomes and portions of the cytoplasmic ground substance; membranes are mostly unmarked.


A simple polychrome stain for conventionally fixed Epon-embedded tissues. The described technique, based upon a one-step Mallory-Heidenhain stain, can be applied as a routine stain for glutaraldehyde or OsO4 fixed, Epon embedded tissues of various organs. The technique consists of a short treatment of the sections with H2O2, a nuclear staining with celestine blue B and a final staining in a modified Cason's solution. The different tissue and cell components are displayed as follows: dark brown nuclei, yellow cytoplasm, red collagen fibers and blue elastic fibers. Intracytoplasmic components as glycogen and mucus are stained respectively blue and violet, whereas other inclusions such as leucocyte granules are colored orange to red.


Dichromatism of bromphenol blue, with an improvement in the mercuric bromphenol blue technic for protein. Staining of protein in sections using the mercuric bromphenol blue technic is improved by staining with 1% HgCl2 and 0.05% bromphenol blue in 2% aqueous acetic acid for 15 min at room temperature. Rinse slides 20 min in 2 changes of 0.5% aqueous acetic acid. Blot and give 2 fast changes in absolute ethanol with agitation before transferring to xylene. Transfer slide to 0.5% n-butylamine in xylene for a few seconds until the section is blue, then, after 2 changes of xylene, mount in DPX. Spectrophotometric analysis of this blue dye at different concentrations and with or without heparin showed that the reddish hues are due to dichromatism and not metachromasia.


An acetylcholinesterase method for in toto staining of peripheral nerves. Stomach, small intestine, uterus, urinary bladder, vagina, mesentery, mesometrium and joint capsule of rats, gall bladder, cystic duct and bile duct of dogs and uteri of young children are stained in toto. Tissue is perfused with saline containing hyaluronidase, then pinned on a flat layer of Paraplast and fixed for 24 hr in cold sucrose formol solution. Stomach, urinary bladder and gall bladder are also fixed in toto. Rinse for 2 days in cold 0.22 M sucrose in a sodium cacodylate buffer pH 7.2. Incubate in medium consisting of 60 mM acetate-buffer pH 5.0 or pH 5.6 (for human material only), 2 mM acetylthiocholine iodide, 15 mM Na citrate, 3 mM Cu sulphate, 0.5 mM K3Fe(CN)6, 5 times 10-4 M iso-OMPA, 1% Triton X 100 at 37C. Rinse in doubly distilled water. Dehydrate in glycerine/water mixtures of increasing glycerine content. Store in glycerine or delaminate under dissecting microscope. Delaminated specimens are mounted on gelatinized object glasses, cleared in xylene and coverslipped with Malinol. Specimens stored in glycerine can be studied microscopically. Stained specimens can also be embedded in Paraplast and sections can be studied after counterstaining.


Suppression of connective tissue impregnation in a silver technique for demonstrating nerve fibers. A tissue pretreatment is introduced which effectively suppresses the silver impregnation of connective tissue and nonspecific background elements in peripheral nerve. The result is a selective impregnation of nerve fibers. The procedure utilizes fresh frozen sections and can be used with the Holmes (1947) or Bodian (1936) techniques. Fresh frozen sections are cut at 10 microns, mounted on slides and air dried for 5 minutes. They are fixed for 30 minutes in formol-sublimate (10% formalin saturated with mercuric chloride) and then placed into 0.5% iodine in 70% alcohol for 5 minutes followed by bleaching in 2.5% sodium thiosulfate for 2 minutes. After washing in running tap water for 10 minutes and a brief rinse in distilled water, impregnation is accomplished by the Holmes (1947) or Bodian (1936) procedure beginning with the step containing the aqueous silver solution. The results show an absence of impregnation of connective tissue and nonspecific background. The technique is simple, rapid, and, by utilizing fresh frozen sections, can be used for other histological and histochemical purposes. Several experiments were done to determine the causes of the connective tissue and background suppression. The air drying step was omitted; the sections were fixed in formalin without mercuric chloride; and the formol-sublimate fixation time was increased. The results suggest that connective tissue impregnation is suppressed by the use of mercuric chloride in the fixative and that the background suppression is related to the short fixation time with formolsublimate.


The use of Herr four-and-a-half clearing fluid for the rapid microscopic examination of thick sections of normal and neoplastic tissues. We have demonstrated that Herr's 4 1/2 clearing fluid, developed for use with plant tissues, can be successfully used for the microscopic examination of thick sections of normal and neoplastic mammalian tissues. Rat Novikoff hepatoma, rat liver, and human colon and skin samples were fixed in Bouin's, stained with iron hematoxylin, treated with Herr's 4 1/2 clearing fluid and examined by phase contrast microscopy. Tissue architecture and cytological detail were easily observed by focusing through tissue sections as thick as 70 mu. The method permits rapid microscopic examination of mammalian tissues and enables the investigator to detect readily morphological abnormalities within a tissue.


A staining method using acridine orange and auramine O for fungi and mycobacteria in bovine tissue. A rapid staining method using auramine O and acridine orange (AOAO) is described for staining mycobacteria and fungi in paraffin sections of bovine tissues. One hundred seventy-seven tissue sections from specimens divided into two general groups on the basis of previous histopathology results were examined with a fluorscent microscope. Group I, a total of 77 sections, were from 47 mycobacterial and 30 fungal granulomas. Mycobacteria were found by the AOAO procedure in 44 of the 47 tissues previously diagnosed as positive for mycobacteria. All 30 fungal granulomas previously diagnosed using convential fungal stains were positive with the AOAO procedure. Group II consisted of sections prepared from 100 granulomas in which typical mycobacterial lesions were observed by histopathologic examination but in which no mycobacteria had been detected. Using the AOAO procedure, two of these 100 granulomas were found to contain mycobacteria and two were found to contain bacterial colonies. In the remaining 96 no etiologic agent could be demonstrated.


Cytological differentiation of asymptomatic pancreatic islet cell tumours in autopsy material. In eleven cases thirteen pancreatic islet cell adenomas were found in autopsy material from 1366 adult cases. Ten of the adenomas were solitary, while 3 small adenomas were observed in a single case. Another four possible solitary adenomas were observed, but their identity was uncertain owing to marked fibrosis. All the adenomas contained A-2 (A)-1 cells but no B (B)-1 cells. Nine of them also contained A-1 (D)-1 cells. The majority of cells in the adenomas were A-2 cells or cells which did not stain with any of the techniques used. The 4 possible adenomas contained islet cells (A-1, A-2, B) in different proportions. With one exception the patients with adenomas and possible adenomas were 65 years of age or older, and in some of these cases adenomas or hyperplasias were also found in other endocrine organs. The frequency of gastroduodenal ulcers or scars in the cases with adenoma or possible adenoma did not differ notably from that found in the cases without pancreatic adenomas. Among the cases with pancreatic adenoma and possible adenoma there were 3 patients with maturity onset diabetes mellitus, but otherwise no clinical symptoms of endocrine disturbances were noted.


HBs antigen in human plasma fractions.

HBs antigen in human plasma fractions. Therapeutic fractions were obtained by fractionating human plasma containing HBs antigen, by the Cohn ethanol technique modified by Nitschmann. Plasmas with various HBs antigen titers were selected. The antigen was sought in each fraction by the techniques of counterelectrophoresis and radioimmunological detection in liquid phase. With the exception of very pure albumins, all fractions were found to contain the HBs antigen, though in variable proportions. A technique for the purification of albumin, applicable on an industrial scale, is proposed. It makes it possible to obtain an albumin fraction in which the HBs antigen cannot be detected by the most sensitive techniques, even starting with a plasma which is very rich in HBs antigen (titer 1/64 by counterelectrophoresis).


C3, GBG, orosomucoid and haptoglobin polymorphisms. Improved staining methods. C3, GBG, and orosomucoid polymorphisms were electrophoresed in a high-voltage agarose method which permitted the typing of 15-20 samples. The increased sensitivity of the dye Coommassie blue was used to stain the protein, and yielded higher resolution than amido black. The typing of haptoglobin samples, was facilitated by devising a method which utilizes the peroxidase activity of the haptoglobin-hemoglobin complex with 90-tolidine and hydrogen peroxide as substrates and 4-chloro-1-naphthol as coupler.


Recent advances in the immunopathogenesis of systemic lupus erythematosus. Systemic lupus erythematosus (SLE) is a chronic multisystem inflammatory disease having definite etiologic associations with ethnic, genetic, viral and immunologic factors. Its pathologic hallmark, vasculitis, is currently felt to be the end result of an immune-complex mechanism. Several clinical and serologic variants of SLE are recognized including discoid lupus erythematosus (DLE), mixed connective tissue disease (MCTD) and drug-induced equivalents-such as procainamide-induced lupus (PIL). The distinguishing features of these variants as well as their prognosis and therapy are discussed in relation to recent developments in the immunopathogenesis of SLE.


Formation of cysteinyldopa from glutathionedopa in melanoma. Glutathionedopa injected intravenously into mice is metabolized and excreted in the urine as a compound with the fluorescence characteristics of cysteinyldopa. Glutathionedopa incubated with a guinea-pig kidney homogenate is metabolized to a compound with the fluorescence characteristics of cysteinyldopa. Boiling of the kidney homogenate prevents the metabolism of glutathionedopa. Incubation of glutathionedopa with a homogenate of a melanoma metastasis led to the formation of a compound with the fluorescence characteristics of cysteinyldopa. Boiling of the melanoma homogenate prevented the metabolism of glutathionedopa. Large amounts of glutathione added to the incubate inhibited the reaction. Lung tissue and blood plasma had no detectable ability to metabolize glutathionedopa. The results show that human melanoma contains one or several enzymes capable of metabolizing glutathionedopa to a smaller dopathioether, probably cysteinyldopa. Such enzymes seem to be normally present in mice and guinea-pigs and have been demonstrated in the guinea-pig kidney.


Cytopathology of nipple discharge due to Pityrosporum orbiculare and cocci in an elderly woman. A case has been described where cytologic examination first showed a fungus infection as the possible cause of a nipple discharge of long duration. The discharge was found to be associated with Pityrosporum orbiculare, Micrococcus tetragenes and Staphylococci (coagulase negative). Administration of an antibiotic that inhibited the bacteria cured the condition.


Slit-scan flow system for automated cytopathology. A low resolution slit-scan flow system is described. Cells flow in solution through a thin "wall" of fluorescence excitation illumination where they are sequentially illuminated. Computer analysis of the resulting slit-scan contours provide a measure of cell diameter, nuclear diameter, nuclear to cell diameter ratio in percent, nuclear fluorescence, cytoplasmic fluorescence, and certain low resolution morphological information. The system, its potential, and remaining problems are discussed.


The sebotrophic effect of pregnancy.

The sebotrophic effect of pregnancy. The sebum excretion rate (SER) in forehead skin was measured in late pregnancy and 8-20 weeks after delivery in 43 women. There was considerable individual variation but the mean SER during pregnancy was significantly higher than in a control group of non-pregnant women and it showed a large and significant decrease to the control level after delivery. This suggests that a powerful sebotrophic hormone is secreted during pregnancy. The mean SER during pregnancy in women with twins or triplets was no greater than the SER in women with a single foetus. This suggests that the sebotrophic factor of pregnancy is unlikely to be of placental origin, and is in keeping with the idea that the sebotrophic factor may come from the pituitary.


A comparative study of the Papanicolaou staining method and the fat stain technique in malignant and non-malignant lesions of the urinary tract. We have collected urine specimens from cases of malignant and nonmalignant lesions of the urinary system and have proceeded to a comparative study of these cases with their clinicopathologic findings after staining them with the Papanicolaou method and the fat stain method. It was found that the fat stain is not specific for renal cell carcinoma only. However, a combination of fat stain and Papanicolaou stain as a second check up of potential malignancy would be the best way of examination.


Immunological phenomena in harmless mice during experimental carcinogenesis induced with long-term topical application of 7, 12-dimethylbenzanthracene (DMBA). To estimate the influence of topical treatment with DMBA and induced tumors on delayed hypersensitivity, the response of spleen lymphocytes to PHA in vitro and macrophage migration inhibition with PHA were studied in DMBA-treated hairless mice. DNA synthesis and blastic transformation of cultured lymphocytes decreased after 6-12 weeks of DMBA application. Lymphocyte response to PHA gradually diminished during the experiment, as compared with control animals. Since the malignant transformation of skin tumors was not observed before 16 weeks of DMBA carcinogenesis, it seems that derangements in cellular immunity preceded the malignant proliferation. The increase in spleen weight and the absence of PHA-induced inhibition of macrophage migration in hairless mice with malignant tumors may also be related to the influence of the tumor itself on the lymphatic system of experimental animals.


Epidermal surface saccharides reactive with phytohemagglutinins and pemphigus antigen. Inorder to study the relationship between epidermal surface saccharides and pemphigus antigen(s), fluorescein-labelled Concanavalin A (Con A) and Phytohemagglutinin-P (PHA-P) were used. These phytohemagglutinins were found to bind with the intercellular areas of human epidermis. Alpha-methyl-D-mannoside and alpha-methyl-D-glucoside inhibited the epidermal intercellular staining pattern produced by ConA-FITC, while N-acetyl-D-galactosamine blocked the same staining pattern produced by PHA-P-FITC. Normal human skin reacted with pemphigus antibody and pemphigus skin with the deposition of IgG both gave a positive intercellular staining pattern with fluorescein labelled phytohemagglutinins. Our data indicated the non-identity of the binding of Con A and PHA-P, and pemphigus antigen(s).


Immunofluorescence studies in herpes gestationis. In 4 cases of H.G., direct junctional immunofluorescence findings showed an intense band of C3 present in the involved skin of the patients even after the active lesions had disappeard (i.e. 2 months later). Weak specific fluorescence for IgG and in one case also for IgA and IgM was seen in the involved skin. A less intense junctional band of C3 was also present in the uninvolved skin. Antibodies against basal membrane have not been detected in the sera of the patients.


Allergens in sesame oil contact dermatitis. In 13 patients with contact allergy to sesame oil, studies were undertaken to elucidate the nature of the allergens. Sesamol, sesamin and sesamolin were identified in crude and purified (pharmaceutical) sesame oil. Patch tests showed 8 of the 13 patients to be positive to sesamol and 12 to sesamolin and sesamin. Patch tests with the pure substances on thin-layer sheets were inconclusive as to any difference between these substances. Group allergy to several substances related to sesamol could not be clearly demonstrated.


Scanning electron microscopy of acarus scabiei. Details in the anatomy of sarcoptes scabiei var. hominis are described. Scanning electron microscopy is of help in illustrating this parasite which is commonly seen in clinical practice. The technique also provides the possibility to differentiate between various types of mite.


Cellular changes in the basaloid cell papilloma. Electron microscopic analyses of basaloid cell papillomas of the solid and papillomatous types are reported. The submicroscopic organization is described. Some of the ultrastructural findings, e.g. an increased number of mitochondria, a certain mitochondrial polymorphismus, the occurrence of irregularly shaped intracytoplasmic vesicles, the abundance of endoplasmic reticulum hypertrophy and the remarkable presence of microtubule-like structures, an unusual finding in a material fixed at the temperature used, are indicating an altered metabolic activity. The alternating presence and absence of keratohyalin is found to be submicromorphologically related to the formation of A- respectively B-cells. This is compared with the formation of parakeratosis in psoriatic lesions without keratohyalin. A formation of orthokeratosis as seen by the light microscopical procedure seems possible without preceeding occurrence of keratohyalin.


5-S-cysteinyldopa in the plasma of melanoma patients and the renal clearance of this amino acid. 5-S-cysteinyldopa has been demonstrated in the plasma of two patients with metastases of malignant melanoma and a high excretion rate of 5-S-cysteinyldopa in the urine. In one patient the plasma clearance of 5-S-cysteinyl dopa was 30 ml/min and in the other 69 ml/min. These clearance values were 43 and 45%, respectively, of the creatinine clearance in the two patients.


Periodic acid methenamine silver (pam) staining of the human subcutaneous lymphatic vessel. The fine structure of the subcutaneous lymphatic vessel taken from the dorsum pedis of the human adult was investigated without and after impregnation by periodic acid methenamine silver (PAM). To check the stainability of another tissue, the renal glomerulus of the rat was also examined in the same way as outlined above. The membrane system of various cells stains positive. No special surface coat of the endothelial cell of the lymphatic vessel was detected. The fine intracellular filament as well as a continuous basal lamina of the endothelial cell were almost PAM-negative, although a part of the basal lamina was occasionally weakly positive. The chemical composition of the basal lamina of the lymphatic vessel is considered to differ from that of the glomerular blood capillary of the kidney, which is PAM-positive. This difference is thought to be due to the smaller polysaccharide content in the former than in the latter.


Histological and histochemical investigations of lupus erythematosus in the skin and the oral mucosa. Skin and oral lesions of chronic discoid lupus erythematosus from 6 patients have been investigated histologically and histochemically. Intra-individual comparisons between skin and oral lesions and intrasection comparisons between clinically affected and unaffected regions were performed. Enzyme histochemical recordings consisted of oxido-reductase and hydrolase activities. A high degree of correlation was noted in the intra-individual comparisons. The intrasection comparisons provided information concerning metabolic dynamics of the disease. The data obtained support the view that serious vascular changes are involved in the development of the changes in the overlying epithelium. The enzyme histochemical results agreed generally with histological data and proved to be a valuable diagnostic aid.


Friction blisters as a manifestation of pathomimia. Self-inflicted skin lesions (pathomimia, artefacts) can be produced in a variety of ways, by the help of nails, razor blades, cigarettes, chemicals, etc. The present study reports three cases of pathomimia, in which the lesions consisted of vesicles or bullae producted by friction. Friction blisters can easily be produced in sites where the epidermis is both thick and firmly attached to the underlying tissues. The blisters have a characteristic histological appearance, which makes it possible to distinguish them from the blisters of different forms of vesicular and bullous diseases and from burn blisters which may also occur in pathomimia.


Excretion of 5-S-cysteinyldopa in the urine of healthy subjects. Seventy-six Caucasians, 30 men and 46 women, were investigated for the 24-hour excretion of 5-S-cysteinyl dopa in the urine during the months of September to November, 1973. No subject had had strong sun exposure for at least 4 weeks prior. A preliminary finding of a variation of 5-S-cysteinyldopa with season necessitated this precaution. The excretion varied between 9.0 and 242 mug/24 hours. The mean value in men was 100 mug/24 hours and in women 77.8 mug/24 hours. Subjects with white hair had lower values than those with pigmented hair, but there was no other difference between the excreted amounts in subjects with differing hair colour. There was no variation with age when the subjects with white hair were excluded. No variation with weight or body surface was found. Excretion of dopa and dopamine determined together did not correlate with the excretion of 5-S-cysteinyldopa.


Surgical treatment of axillary hyperhidrosis in 123 patients. 123 patients suffering from axillary hyperhidrosis have been operated on as outpatients under local anaesthesia, with radical ablation of sweat glands. All except 7 patients were satisfied with the result. There were few complications. A group of patients and controls were extensively studied including quantitative sweat testing and axillary biopsis. Apart from increased axillary sweating during the test, no differences could be found. The basic etiology of the condition is still unknown.


Interaction of contraceptive progestins and related compounds with the oestrogen receptor. Part I: Effect on (3H)oestradiol distribution pattern in the ovariectomized rat. The distribution pattern of oestradiol in ovariectomized rats as a function of time has been studied following intravenous adminstration of the tritiated hormone. Oestrogen specific binding with limited capacity was observed in the uterus, vagina, anterior pituitary, adrenals, preoptic area, hypothalamus, amygdala, septum and tractus diagonalis. Maximal uptake of oestradiol in the pituitary occurred within 5 min, in the uterus 60 min after injection, and remained almost unchanged at this level for more than two hours. The binding capacity per mg tissue decreased in the order pituitary, uterus, vagina, preoptic area, adrenals, hypothalamus, amygdala, spetum and tractus diagonalis. The hormone concentration in these tissues one hour after (3H)oestradiol injection was lowered by previous administration of ethinodiol, norethinodrel, lynestrenol and norethindrone, whereas medroxyprogesterone, chlormadinone, megestrol and methyllynestrenol had no effect. The same results were obtained, when instead of the steroid alcohols the corresponding acetate esters were administered. For norgestrel, oestrenol and nortestosterone the effect in the dose range studied was limited to the pituitary and preoptic area. For lynestrenol the inhibition of oestradiol binding in the target tissues was almost the same when the progestin was given 60 and 5 min before oestradiol, whereas in the case of administration 30 min after oestradiol no inhibition was observed. The reduction of oestrogen binding appeared to be dose-dependent, but the dose required to obtain a certain effect for the uterus was four times as high as for the pituitary. Discrepancies between previous studies and the implications of the present findings for the mechanism of action of ovulation inhibition by these progestins are discussed.


Interaction of contraceptive progestins and related compounds with the oestrogen receptor. Part II: Effect on (3H)oestradiol binding to the rat uterine receptor in vitro. Incubation of the 105 000 times g supernatant of rat uterus homogenate with (3H)oestradiol resulted in an oestrogen specific binding of limited capacity to a macromolecule sedimenting in the 8-9S region after density gradient centrifugation. The contraceptive progestins of the 19-nortestosterone series were able to interfere with oestradiol binding in contrast to the hydroxyprogesterone derivatives chlormadinone, medroxyprogesterone and megestrol. The interaction appeared to be competitive. The strongest inhibition of oestradiol binding was observed in the presence of ethinodiol, followed by northinodrel, lynestrenol and norethindrone respectively. Norgestrel was almost inactive. Of the related structures tested oestrenol displayed more activity than norethindrone, nortestosterone and ethisterone were less active and 6alpha-methyllynestrenol showed only border line activity. In comparison with norethinodrel and norethindrone, lynestrenol and oestrenol appeared in vitro to be stronger competitors for oestradiol than in vivo (Part I; Van Kordelaar et al. 1975). This may be due to the great difference in lipophilic character, which is reflected in the RM values of these compounds. From the results obtained it may be concluded, that the presence of a 17alpha-ethynyl substituent promotes receptor binding, whereas the introduction of methyl substituents in the position 6, 10 and 18 causes the opposite effect. The relationship between the various ring A structures and the affinity to the receptor is discussed.


A reassessment of the distribution of multiple sclerosis. Part one. When reviewed some 10 years ago, available prevalence studies of multiple sclerosis (MS) seemed to divide the world into three frequency zones for MS: high prevalence at 30 to 60 per 100,000 population; medium at 5 to 15; and low at less than 5 per 100,000. In the last decade the number of the available studies has more than tripled. Their reassessment, including judgments of comparability, still indicates a high-medium-low division of MS frequency world-wide. The risk areas comprise northern Europe, northern United States, much of southern Canada, New Zealand, and probably southern Australia. Prevalence rates in these regions are mostly 30 to 80 per 100,000 population, centering at about 50. Medium frequency is defined as prevalence of 5 to 25, and is mostly 10 to 15. In Europe, the medium frequency zone bounds that of high frequency to the north, east, and south. The European Mediterranean basin is of medium prevalence with a sharp division from the high zone across France and Switzerland. It is likely that this division continues eastward across Austria, north of Hungary, and across the upper Ukraine to the Caspien Sea, but this is not definite. Medium risk areas of Europe thus include surveyed sites of Spain, Italy, Hungary, Jugoslavia, Bulgaria, and central Ukraine, together with southeastern France and southern Switzerland. Though Romania could be high, it is more likely to be of medium prevalence. Turkey measures low, but from incomplete data. From nationwide prevalence and mortality studies, the west coast of Norway and all Scandinavia above latitude 65 degrees north are of medium frequency. Based on hospital data, northwestern USSR is high, and central and southern USSR medium, in MS risk. Other medium risk areas include southern United States, most of Australia, one ethnic group only in South Africs, and possibly Hawaii. Low risk areas are allsurveyed sites of Asia, the Pacific islands, Africa, Latin America, Alaska, and Greenland.


Clues to cancer risk: biologic markers.

Clues to cancer risk: biologic markers. Two large kindreds manifesting the "cancer family syndrome" have been studied. Genetic and biologic studies reveal markers which have important implications for cancer detection, control and prevention. Valuable markers may be found in the major histocompatibility system, HL-A. The presence of carcinoembryonic antigen and SV-40 viral transformation of skin fibroblasts are other possibly important markers. Relationships of the ABO blood groups to cancer are under intensive study, as is the hepatitis-associated antigen.


Morphological and histochemical studies on a PAS-positive granular leukocyte in blood and connective tissues of Catostomus commersonii Lacépède (teleostei:pisces). This study was undertaken to identify an ubiquitous granular leukocyte found in Catostomus commersonni Lacépède. The cell contains large, numerous, strongly PAS-positive cytoplasmic granules, an eccentric nucleus and prominent, persistent juxtanuclear space. It develops in the hemopoietic tissue of the kidney, and mature cells are found not only in kidney and peripheral blood but also in areas of connective tissue where mast cells are usually located. Electron microscopy confirms the presence of a large Golgi apparatus, unlamellated cytoplasmic granules and extensive rough-surfaced endoplasmic reticulum. Histochemical studies show that the cytoplasmic granules are alcianophobic, non-metachromatic and unstained by acridine orange. Histamine is detectable spectrophotometrically in kidney tissue, but the PAS-positive granular leukocyte does not consistently degranulate after treatment with histamine liberator 48/80. The authors suggest that while the PAS-positive granular leukocyte is not identical with classical basophils/mast cells, which are absent in C. commersonnii, it may represent an evolutionary precursor of these cells.


Preparation and standardization of perotidase-labeled anti-human IgG antibody for use in determination of serum antinuclear antibody levels. Peroxidase-labeled antibody conjugates were prepared by a two-step conjugation procedure using glutaraldehyde. Immunoadsorbent-purified antibody and the gamma-globulin fraction of sheep anti-human IgG antiserum were employed for these preparations, Procedures for the determination of the antibody and enzymatic activity, as well as the specificity of the enzy-e-antibody reactions, were outlined. Peroxidase-anti-human-IgG conjugates were prepared with approximately a 1:1 molar ratio of peroxidase to IgG, which demonstrated a loss of 10-15% of precipitating antibody activity. Standardization procedures for use of peroxidase-labeled antibody in the indirect ANA test were established. The peroxidase-labeled antibody in the indirect ANA test were established. The peroxidase-antibody procedure was found to demonstrate a reproducible plateau endpoint which was comparable to that of the fluorescent antibody preparations.


The identification of lupus erythematosus cells. A comparison of supravital fluorochromation with a standard Wright's stain method. A comparison of standard Wright's stained lupus erythematosus preparations and an acridine orange fluorochromatic method was conducted using 354 consecutive lupus erythematosus preparations involving 264 patients. The results of this comparison and a discussion of the fluorochromatic procedure are presented.


Limitations of the usefulness of the d-xylose absorption test. The results of the conventional 5-hour d-xylose absorption test were surveyed in 38 subjects with disease of the jejunal mucosa, giardiasis or bacterial overgrowth, or no small-bowel disorder. The test was in error in 20-40% of cases, depending on the disease category, and the error for the entire group was 30%. We conclude that the test yields little guidance for diagnosis or therapy of clinical problems and is superfluous when a jejunal biopsy can be obtained.


Periodic acid-Schiff-light green stain to detect glomerular protein deposits by routine light microscopy. A simple, easily reproducible periodic acid-Schiff-light green stain (PAS-LG) for the detection of glomerular protein deposits by routine light microscopy is described. The deposits are selectively stained a deep blue and contrast sharply with the staining of adjacent glomerular structures. Correlation with immunofluorescent and electron microscopy has shown that it is possible with this stain to categorize accurately a large variety of glomerular lesions by light microscopy alone.


Post-jejunoileal-bypass hepatic disease. Its similarity to alcoholic hepatic disease. The authors studied serial hepatic biopsies of five patients who developed hepatic failure following jejunoileal bypass for extreme obesity, with autopsies of two. The hepatic histologic changes included centrilobular or focal alcoholic hyalin, intrasinusoidal collagenosis, fatty hydropic degeneration, and neutrophilic infiltrate. At least two of the patients were abstinent from alcohol, both prior to and after the surgical procedures. The others, after the bypass procedures, had reduced alcohol consumption from previous levels. All patients developed hepatic failure and histologically progressive hepatic disease with alcoholic hyalin and other changes indistinguishable from alcoholic hepatic disease in 21/2 to 5 months, in spite of hyperalimentation and re-establishment of intestinal continuity in four. Nausea, vomiting, abdominal pain and ascites were prominent complaints. Four of the five patients died in hepatic failure. The authors conclude that these cases of progressive hepatic disease with histologic changes simulating those found in livers of alcoholic patients offer evidence that heavy alcohol consumption may affect the liver in an indirect fashion.


Immunologic and cytochemical properties of histiocytic and mixed histiocytic-lymphocytic lymphomas. Lymph nodes were biopsied from seven patients with the presumptive clinical diagnosis of lymphoma and studied for their ability to form spontaneous rosettes with sheep erythrocytes (T cell marker), for surface immunoglobulins (B cell marker), for cytochemical reactivity with peroxidase, alpha-naphthyl acetate and butyrate esterases, naphthol ASD chloroacetate esterase, acid phosphatase, periodic acid-Schiff, Sudan black B, and Wright-Giemsa on touch preparations, as well as in hematoxylin and eosin-stained sections. Lymph nodes from patients without hematologic malignancy served as control. Diagnoses of diffuse histiocytic lymphoma were made in five cases and diffuse mixed histiocytic-lymphocytic lymphoma in 2 cases. The cytochemical staining of the lymphoma cells were typical of lymphoid cells rather than macrophages. In five cases neoplastic cells contained surface immunoglobulins, suggesting a B cell origin, and in one case a paucity of cell surface markers was found. Cells from uninvolved nodes of lymphoma patients could not be differentiated from those of the control group.


Acute lymphocytic leukemia with atypical cytochemical features. In a series of 78 cases of acute lymphocytic leukemia, seven were found to demonstrate blast cells staining strongly for both the periodic acid-Schiff reaction and alpha-naphthyl acetate esterase. It is suggested that these features may represent a subclassification of acute lymphocytic leukemia or may be a marker for the leukemic phase of lymphoma.


Gamma globulin complexes in rheumatoid pericardial fluid. Cardiac tamponade due to pericarditis occurred in a patient with rheumatoid arthritis. Aspiration afforded us an opportunity to expand the characterization of pericardial fluid. Elevated acid phosphatase levels, decreased whole hemolytic complement and gamma globulin complexes similar to those found in rheumatoid synovial fluid were noted, supporting the concept of a unitary nature of inflammation in rheumatoid disease.


Incidence and description of accelerated ventricular rhythm complicating acute myocardial infarction. One hundred and nineteen episodes of accelerated ventricular rhythm (less than 125/min) were noted in 37 patinets with acute myocardial infarction during a 1 year period. The incidence was 12.7 per cent. Twenty-seven episodes of fast ventricular tachycardia (less than 125/min) were noted in 16 of these patients. Eighteen patients had anterior myocardial infarction and 19 inferior myocardial infarction. The mechanism of onset of accelerated ventricular rhythm was classified as escape in 65 episodes. Ventricular premature beats were noted close to episodes of accelerated ventricular rhythm in 31 patients and fast ventricular tachycardia in 14 patients. The morphology of accelerated ventricular rhythm was similar to the ventricular premature beats in 27 patients and similar to the fast ventricular tachycardia in 12. In 11 patinets the morphology of ventricular premature beats, accelerated ventricular rhythm and fast ventricular tachycardia were all the same. In six patients the coupling time of the ventricular premature beats and the onset of the accelerated ventricular rhythm were the same. In seven patients the morphology of the accelerated ventricular rhythm and fast ventricular tachycardia were the same, and the rate of the accelerated ventricular rhythm was exactly half that of the fast ventricular tachycardia. There were three deaths due to shock and heart failure. Three episodes of fast ventricular tachycardia progressed to ventricular fibrillation and were successfully cardioverted. It is concluded that accelerated ventricular rhythm and fast ventricular tachycardia were all the same. In six patients the coupling time of the ventricular premature beats and the onset of the accelerated ventricular rhythm were the same. In seven patients the morphology of the accelerated ventricular rhythm and fast ventricular tachycardia were the same, and the rate of the accelerated ventricular rhythm was exactly half that of the fast ventricular tachycardia. There were three deaths due to shock and heart failure. Three episodes of fast ventricular tachycardia progressed to ventricular fibrillation and were successfully cardioverted. It is concluded that accelerated ventricular rhythm is a relatively common complication of both anterior and inferior myocardial infarction. The high incidence of concomitant fast ventricular tachycardia, the frequency of ventricular premature beats with similar morphology and coupling time, and the instances of two arrhythmias having common rate multiples, suggest that at least in some instances accelerated ventricular rhythm may represent an ectopic focus with exit block.


Uncontrolled diabetes mellitus and hyperglucagonemia associated with an islet cell carcinoma. A 53 year old woman presented with diabetes mellitus, hyperglucagonemia (600 to 1,500 pg/ml), clinical hyperparathyroidism and an abdominal mass diagnosed on biopsy as an islet cell carcinoma. Glucagon content of the tumor was 0.78 mug/g wet weight. Hourly blood samples during a 24 hour period revealed a direct correlation between plasma glucose and glucagon. The oral administration of glucose paradoxically increased whereas the intravenous administration decreased plasma glucagon. Circulating glucagon levels were markedly increased with arginine and epinephrine infusion. Both short- and long-term administration of alpha adrenergic blockade depressed the glucagon response to epinephrine infusion. In contrast, long-term alpha adrenergic blockade increased glucagon secretion despite improved glucose tolerance during a second 24 hour study. Although the patient demonstrated overt clinical and chemical findings of hyperparathyroidism, parathyroid hormone (PTH) was not detected in her plasma. The pattern of tumor growth was consistent with an origin from pancreatic islets. We conclude that (1) the tumor was responsive to physiologic stimuli known to affect glucagon secretion; (2) elevations of plasma glucagon levels with oral and dietary glucose suggest regulation of secretion by intestinal factors; and (3) improvement of glucose tolerance with alpha adrenergic blockade may be related to increased insulin secretion.


Nephropathy associated with sickle cell anemia: an autologous immune complex nephritis. I. Studies on nature of glomerular-bound antibody and antigen identification in a patient with sickle cell disease and immune deposit glomerulonephritis. The nature of the glomerular-bound antibody and the putative antigen was investigated in one of the patients with sickle cell disease and immune deposit membranoproliferative glomerulonephritis by immunohistologic and glomerular antibody elution. Renal proximal tubular epithelial antigen was localized in association with immunoglobulins G (IgG), M (IgM), Clq fraction of the first component of complement (Clq) and the third component of complement (C3) in a granular pattern along the glomerular basement membrane of the patient's kidney. IgG and IgM were eluted from glomeruli. These immunoglobulins fixed to the proximal tubules of normal human kidney by direct immunofluorescence. This localization was abolished by absorption of the eluted immunoglobulins with renal tubular epithelial (RTE) antigen. The IgG eluted from the glomeruli blocked the fixation of rabbit anti-RTE antigen to normal proximal tubular brush border. These studies suggest that the nephritis in this patient was due to deposition of complexes or RTE antigen and specific antibody. An autologous immune complex nephritis may develop in some patients with sickle cell anemia secondary to RTE antigen released possibly after renal ischemia or some other phenomenon causing renal tubular damage.


A comparison of the Murphy-Pattee and the Seligson methods for serum thyroxine. We compared two methods for serum thyroxine measurement by competitive protein binding - the Murphy and Pattee and the Seligson and Seligson methods. We found the Seligson and Seligson method, which requires less sample volume and analysis time, to be the method of choice on the basis of sensitivity, extraction efficiency, precision, and accuracy. Standard plots of time/10,000 counts versus thyroxine concentration are linear to 20 ug/dl, extraction efficiency is 99.6 per cent, within-day S.D. plus or minus 0.28 ug/dl, and 98.3 per cent of added thyroxine is recovered in the Seligson and Seligson method. The Seligson and Seligson method is superior to the Murphy and Pattee method with respect to all these parameters. The methods were also compared with correlation and normal value studies.


An electron microscopic study of fetal capillary basal laminas of "normal" human term placentas. Electron microscopic examination of terminal (free-floating) villi of definable normal human term placentas emphasizes the variable fine structural appearance of the fetal capillary basal lamina (FL). The FL assumes both a unilaminar and multilamellar configuration in each placenta examined; the multilamellar portions are usually composed of individual layers which tend to branch and anastomose. The constant indentification of collagen fibrillar profiles between these electron-dense layers argues against the notion that the multilamellar portions of the FL arise via a simple splitting or disaggregation of pre-existing unilaminar FL's.


Effects of human chorionic gonadotropin preparations on complement in vitro. The effects on complement in vitro by several commercial preparation of human chorionic gonadotropin (HCG) were investigated. These preparations were demonstrably anticomplementary and produced marked conversion of the third component of complement (C'3). However, detailed analyses showed that the anticomplementary activity was because of the presence of immunoglobulins in the HCG preparations. In view of the known modifying effects of C'3-reactive agents on skin allograft survival and of antigen-antibody complexes on lymphocyte transformation, it may be necessary to reconsider previous work on the immunosuppressive effects of HCG preparations.


Primary hepatic malignancy in pregnant women. Three women dying from hepatic carcinoma during pregnancy are presented. One of these women with a hepatocellular carcinoma and alpha fetoprotein in the serum and antibody to hepatitis B antigen. A fourth patient died 2 months post partum with a cholangiocarcinoma. A false positive pregnancy test suggested that she had metastatic choriocarcinoma in the liver, and a panhysterectomy was performed. The clinical diagnosis with the use of alpha fetoprotein and chorionic gonadotropin for detection of hepatoma and the etiopathogenesis of primary hepatic malignancy in pregnancy are discussed.


Endometrial morphology in women exposed to uterine systems releasing progesterone. A blind study was done in 402 endometrial biopsies of women who had intrauterine devices releasing different daily amounts of progesterone or a placebo (empty device): 175 were obtained in what presumably was the proliferative phase and 227 in the secretory stage, as all these subjects were normal, healthy women with a history of previous fertility. With all dose levels of the progesterone-releasing devices there was variation of the endometrium general pattern and the over-all picture varied from normal secretory to suppressed endometrium. In addition to these changes of the endometrial pattern, in 231 specimens there was significant inflammatory infiltration and in six cases even plasma cells were seen. Predecidual reaction was frequently seen and in 45 cases it was diffuse and marked. The significance of these data is discussed on the grounds of the frequent similarities of the changes here reported with those in women using combined oral steroids for contraception.


Antibodies against spermatozoa in infertile women with poorly invading spermatozoa on postcoital tests. Sixty-four women with poorly invading spermatozoa on postcoital tests (PCT's) were investigated for serum spermatozoal antibodies by the immunofluorescent technique. Control groups tested were women with explained infertility but normal PCT's, normal pregnant women, and normal previously pregnant women. A positive immunofluorescent test was obtained in 19 of 64 women with abnormal PCT's compared with 26 of 51 patients with explained infertility, including 11 of 13 who had well-identified gynecological disorders, 12 of 61 pregnant women, and one of 59 previously pregnant normal women. High titers (larger than or equal to 1/200) were found in a few patients in all groups except normal nonpregnant women. In no patient was evidence of complement fixation found. The immunofluorescent test, as carried out here, is therefore of little value in discriminating a group of patients in whom immunologic abnormalities are the primary cause of the infertility.


Prediction of fetal outcome in threatened abortion by maternal serum placental lactogen and alpha fetoprotein. Abnormally low human placental lactogen (HPL) or high alpha fetoprotein (AFP) levels in maternal serum are unfavorable prognostic signs in women with threatened abortion but normal levels cannot be used to discriminate between viable and nonviable pregnancies. Out of 112 women with threatened abortion, 69 aborted; of these, 36 had a low HPL level and they all aborted. Five women had an increased AFP concentration. Four of these aborted and the remaining case was a twin pregnancy in which one fetus died and the other survived. HPL and AFP levels provide complementary information as to the fetal outcome in threatened abortion. This was indicated by a normal HPL level in all of the five cases with raised maternal AFP, and by a normal AFP level in 35 of the 36 women with low maternal HPL.


Changes in the ratio between serum and "specific" levels of human chorionic gonadotropin in different trimesters of pregnancy. Sixty-one sera from different trimesters of pregnancy were analyzed by two 125-I-NIH-HCG assay systems, employing anti-intact HCG serum and anti theta-HCG serum, respectively. The ratios of the levels measured in the two assay systems changed with the duration of pregnancy. The ratios during trimesters 1,2, and 3 were 2.94, 1.99, and 2.37, respectively. The cross-reactivity of proteohormones other than HCG was tested in both the assay systems. The two assay systems could be comparable in their high degree of specificity. However, the relative affinities of intact HCG and theta-HCG in the two assay systems were observed to be different. It was suggested that the significant differences in the ratios of the levels measured by the tow assay systems might have been infulenced by the occurrence of theta-HCG in serum and that the levels of the subunit must have changed at different stages of pregnancy.


Monitor of chemotherapy in gestational trophoblastic neoplasm by radioimmunoassay of the beta-subunit of human chorionic gonadotropin. Radioimmunoassays and bioassays based on the reactions of the native molecule of human chorionic gonadotropin (HCG) fail to differentiate HCG from pituitary luteinizing hormone (LH). An assay based on the beta-subunit of HCG which detects HCG exclusively has been used in our laboratory to monitor patients undergoing chemotherapy for gestational trophoblastic disease (GTD). We have been able to differentiate minimal, persisting tumor activity from normal levels of pituitary gonadotropins and have based therapy on these findings. Alternatively, treatment has been terminated when HCG is no longer detectable in the serum. Tumor activity has been detected in the beta-subunit assay at a time when biologic activity in the urine indicated remission.


Educational methodology in an obstetric and gynecologic medical school curriculum: a comparison of self-instructional with traditional methods. Five successive groups of third-year medical students were identically tested on the subject "diagnosis of early pregnancy" at the end of their course in obstetrics and gynecology. The first group served as control, receiving no specific instruction on the subject. The next four groups did receive instruction on the subject and in the following ways: tape-slide program, lecture, instructional objectives alone, and instructional objectives plus the same tape-slide program. The average examination scores with S. D. were 40 plus or minus 11, 68 plus or minus 15, 48 plus or minus 17, 42 plus or minus 14, and 83 plus or minus 20 respectively. Analysis of these results reveals that the tape-slide program produced significantly better scores than an identical lecture or the objectives alone, but that accompanying the tape-slide program with the objectives maximized learning. Ramifications and causes of these findings are discussed in detail.


Myocardial conduction defects in association with compression of the umbilical cord. Experimental observations on fetal baboons. Experiments have been performed in 16 pregnant baboons to study in greater detail the fetal hemodynamics, heart rate, and acid-base changes during occlusion of the umbilical cord. A total of 32 observations were made during which the umbilical cord was completely occluded with the fetus intact in utero; 11 were made after the intravenous administration of atropine, 0.01 to 0.02 mg. per kilogram, to fetus. Various degrees of conduction defects occurred in 17 of 21 observations before atropine. Complete atrioventricular block with extrasystoles was seen six times. Upon release of cord occlusion, there was a rapid recovery of myocardial conduction. Atropine prevented myocardial conduction defects; bradycardia was less and the onset was delayed. These observations suggest that myocardial conduction defects which occur during the course of cord occlusion are due to parasynpathetic stimulation. They could also be due to an increased sensitivity of the fetal myocardium to acetylcholine under hypoxic conditions. No recommendation can be made at the present time with regard to the administration of atropine to the fetus when cord compression is suspected on clinical grounds.


Salzmann's nodular degeneration of the cornea. Eleven corneal specimens from nine patients with Salzmann's nodular degeneration of the cornea, together with all available clinical information, were collected for this study. The specimens were examined by light and electron microscopy. An antecedent keratitis was diagnosed by history and microscopic findings in every case. The corneal epithelium showed degenerative changes, its thickness varied, and in nodular areas it often consisted of only a single layer of flattened epithelial cells by light microscopy. Bowman's membrane was missing over the nodules, and in this zone there was excessive secretion of a basement membrane-like material. Hyaline degeneration of collagen, cellular debris, and electron-dense hyaline deposits were seen in the collagen of the nodules. The number of fibrocytes in the nodules varied from many that were active to a few that were degenerating. External irritation because of poor epithelial protection was interpreted as a causative factor, although other tissue repair mechanisms may also have played a role.


Krukenberg's spindles.

Krukenberg's spindles. Forty-three patients who initially presented with Krikenberg's spindles but with no glaucomatous field defects were observed for an average of 5.8 years in an attempt to evaluate the prognostic implication of spindle presence. Twenty-two of the patients had been followed for a minimum of five years. Analysis of the study population revealed that there were more women than men, (26 vs. 17), the women were older than the men, and more women had positive glucose tolerance tests (25% vs. 0). Only two of the patients, both women, developed glaucomatous field defects during the period of observation. Thus, the presence of a Krukenberg's spindle in a patient with no visual field defect is not a particularly ominous sign.


Subconjunctival and episcleral lipid globules. Asymptomatic peripheral subconjunctival and episcleral clear, round globules, 30 to 50 mu in diameter, were found in more than 50% of patients over 50 years of age. This entity occurred in two basic patterns, adjacent to deep conjunctival or episcleral blood vessels or less commonly in isolated patches. These globules increased linearly with age, were more common in women and blacks, with a peak incidence in blacks during the fourth, and in whites, the sixth decade. Histologically, the globules were connective tissue fat cells identical to those found in orbital fat. This entity is not related to obesity, but has a clear relationship to increasing age. The clinical importance of these cells is unknown.


Genetic studies in the Markham Valley, northeastern Papua New Guinea: gamma globulin (Gm and Inv), group specific component (Gc) and ceruloplasmin (Cp) typing. Genetic studies in the Markham Valley, northeastern Papua New Guinea; Gamma globulin (Gm and Inv), group specific component (Gc) and ceruloplasmin (Cp) typing. M. S. Schanfield, Eugene Giles and H. Gershowitz, Department of Human Genetics, University of Michigan, Ann Arbor, Michigan 48104. Immunoglobulin allotyping was carried out on 680 serum samples from inhabitants of the Markham Valley, Papua New Guinea (seven villages speaking the same Melanesian [PAP] speaking village). Family and population data verified the presence of Gm-ag, G-ab and Gm-afb among the MN speakers and Gm-ag, Gm-axg, Gm-ab and Gm-afb among the PAP speakers. The frequency of Gm-ag was between 0.048 and 0.235, while the frequency of Gm-ab was between 0.427 and 0.627 and the frequency of Gm-afb ranged between 0.261 and 0.424 among the seven MN villages; the single PAP village had frequencies of 0.568, 0.160, 0.213 and 0.059 for Gm-ag, Gm-axg, Gm-ab and Gm-afb respectively. The frequency of Inv1 ranged between 0.034 and 0.095 in the MN villages and 0.014 in the PAP village. The rare occurrence of Gm(x) without Gm(g) was explained by the presence of a Gm-axfb haplotype, while in two PAP families the presence of Gm(x) without Gm(g) was explained by the abnormally weak expression of Gm(g) in a Gm-axg haplotype. A total of 654 sera were typed for Gc, with the seven MN villages ranging between 0.350 and 0.650 for Gc-1, 0.312 and 0.575 for Gc-2 and between 0.017 and 0.112 for Gc-Ab; the single PAP village had a value of 0.627 for Gc-1, 0.165 for Gc-2 and 0.208 for Gc-Ab. A total of 693 sera were tested for ceruloplasmin type. All showed the common Cp(b) phenotype.


Alpha-M-fetoprotein in serum of rats after experimental manipulation in the oral cavity. Alpha-M-fetoprotein (AMFP) was detected in the serum of rats subjected to intraoral manipulations, i.e., instillations into the parotid gland and injections into the gingival mucosa of saline or foreign proteins. The identity of AMFP with acute-phase protein is pointed out.


The roentgenographic and clinical findings in Whipple's disease.  A review of 8 patients. Out of 8 patients with Whipple's disease, 7 had roentgenographic findings consisting of slight dilatation of the small bowel, with thickening of the mucosal folds.  The eighth patient had no abnormality on roentgenographic study.  This series represents the first in which the roentgenographic changes have been quantitatively assessed.  Confirmation of the diagnosis by biopsy is important because antibiotic treatment results in marked improvement both clinically and roentgenographically.


Cellular and regional localization of pentobarbital-2-14C by radioautography of selected areas of mouse brain at loss and return of withdrawal response. Regional and cellular distribution of pentobarbital-14C in mouse brain was determined by frozen-section radioautographic methods. The mice were studied at the times of loss (WRL) and return (WRR) of withdrawal response following a single intravenous dose of either 40 or 50 mg/kg body weight. At WRL, grey matter areas had higher concentrations of pentobarbital-2-14C than white matter. At WRR grey matter concentrations were not altered, but white matter areas were now similar to the grey. At WRL pentobarbital concentration was 55 per cent higher in large pyramidal cells in the parietal cortex than in surrounding neuropil. At WRL hippocampal pyramidal cell bodies (stratum pyramidalis) and glial cells in corpus callosum had pentobarbital levels similar to that of surrounding neuropil. Levels in the neuropil of these three areas were higher at WRR than at WRL. Lipid-rich compartments had higher pentobarbital concentrations at WRR than at WRL. The results suggest that return of consciousness after pentobarbital anesthesia is associated with intracerebral redistribution of pentobarbital even while there is continuing uptake into brain. (Key words: Brain, pentobarbital uptake; hypnotics, barbiturates, pentobarbital; pharmacokinetics, pentobarbital uptake.).


Repeated oral administration of coumaphos in sheep: interactions of coumaphos with bishydroxycoumarin, trichlorfon, and phenobarbital sodium. Interactions between treatments with coumaphos, bishydroxycoumarin (an anticoagulane), trichlorfon (an organophosphorous compound), and phenobarbital sodium (an inducer of microsomal enzymes) were investigated in sheep. A daily dose of 2 mg of coumaphos/kg of body weight for 6 days did not affect the plasma enzymes or the antiprothrombinemic effect of bishydroxy-coumarin in wethers. The treatment of ewes with an intravenous (IV) injection of trichlorfon, insufficient to produce significant inhibition of erythrocyte acetylcholinesterase (AChE) activity, appeared to produce additive effects with those produced by subsequent treatment with 4 mg of coumaphos/kg/day. In ewes given 40 mg of phenobarbital sodium/kg for 5 days intraperitoneally (IP), the anticholinesterase effect of 4 mg of coumaphos/kg was significantly reduced and signs of toxicity were not present. Treatment with daily doses of 2 mg of coumaphos/kg for 6 days did not modify the anticholinesterase effect of a 2nd series of treatments given 6 weeks later.


Inherited C2 deficiency and systemic lupus erythematosus: studies on a family. A patient is described in which an inherited defect in the synthesis of C2 complement component coexisted with the disease systemic lupus erythematosus. The family studies show evidence of the autosomal recessive nature of the inheritance of the C2 synthesis defect. Of particular interest was the finding of a great-aunt who also had homozygous C2 deficiency. This great-aunt suffered from discoid lupus erythematosus as well. The occurrence of various autoantibodies in the serum from the family members, the typing for blood groups, HL-A antigens, and some serum protein markers are reported and discussed. The C2 deficiency may be a critical defect in the host defenses to infection that predisposed to the development of autoimmune disease.


Affinity of cellular constituents of two bacteria for fluorescent brighteners. Two fluorescent brighteners were used to stain an isolate of Bacillus cereus var. mycoides and soil pseudomonad.  The stained organisms were fractionated by two procedures to determine which cellular constituents were reacting with the brighteners.  Both fractionation procedures provided evidence that the brighteners were adsorbed to proteins within the cells.  Microscopy examination of ghost cells of the bacillus showed that cell walls were not being stained.  Spheroplasts of the bacillus and the pseudomonad were stained by the brighteners.


[The cell wall of Coelastrum (Chlorophycees)]. The cell wall of Coelastrum is usually composed of three layers. The outermost layer was studied most extensively. It consists of erect tubules which often bear long bristles whose function may be to stabilize the algae in its enviroment. The cell wall can modify its morphology according to the enviroment.


Bilateral adrenalectomy for metastatic breast carcinoma. Ninety-five patients who underwent bilateral adrenalectomy for metastatic breast carcinoma are reviewed. An objective remission of tumor was observed in 66% of the patients over the age of 45 years with metastasis limited to soft tissue, osseous, or pulmonary system. In those patients below the age of 45, only 19% had remission irrespective of the sites of metastasis. Thirty-three patients were admitted with no "free interval," with objective remission occuring in 42%. This observation indicates that the "free interval" is not as striking a determinant in selecting adrenalectomy candidates as had been stressed by others. The serial treatment of adrenalectomy followed by chemotherapy after adrenalectomy failure or relapse was shown to objectively benefit 74% of 72 patients evaluated. Sulfokinase activity in breast cancer tissue was studied in 17 patients. The results were not as definitive as reported by others.


Hemipelvectomy.

Hemipelvectomy. Hemipelvectomy was performed in 50 patients with malignant neoplasms of the upper part of the thigh and pelvis. Although not technically difficult, the operation is associated with considerable blood loss. Postoperative complications are frequent, the greatest morbidity resulting from skin flap necrosis. Symptomatic phantom limb is as occasional late problem. Of 37 patients resected for cure prior to 1969, 14 were alive five or more years postoperatively. Five of these long-term survivors subsequently died of metastases. Patients with fibrosarcoma and chondrosarcoma had the best survival. Six other patients underwent palliative hemipelvectomy for intractable pain, with gratifying results. Hemipelvectomy is an important, useful operative procedure in selected patients.


Corneal elastosis. Appearance of band-like keratopathy and spheroidal degeneration. Clinical and pathologic studies of six corneas from patients with band-like keratopathy and spheroid degeneration were performed. These included cases of noncalcific band keratopathy; in one case very early changes in Bowman membrane were demonstrated by light and electron microscopy. Two other corneas displayed more advanced degeneration that manifested clinically as irregular golden yellow plaques. One case of corneal spheroid degeneration was associated with lattice dystrophy of the cornea. The corneal deposits may be best characterized as a form of elastotic degeneration and were associated with intense autofluorescence.


Protein overload nephropathy in rats with unilateral nephrectomy. A correlative light immunogluorescence and electron microscopical analysis. We wish to determine what cellular and functional alterations are associated with the development of glomeruloscierosis when rats with one kidney are fed an excess of salt or protein. Rats with one kidney are more likely to develop pronteinuria and glomerulosclerosis than control animals. Blood pressure recordings indicate that proteinuria and glomerulosclerosis occur before hypertension is evident. Fluorescent antibody studies disclose that albumin accumulates in the epithelial cells of glomeruli and tubules. Ultrastructural examination shows that vacuolozation of epithelial cells and basement membrane thickening precede the sclerotic collapse of capillary loops. Increased concentrations of sodium or urea that are found in urines of these rats favor the point of view that an elevation of solute load when combined with a reduction of renal mass will on some unknown manner accelerate the deterioration of glomeruli.


Reaction of cultured adult and fetal lung to prednisolone and thyroxine. Pulmonary explants from adult mice were cultured for periods up to six days in medium with and without one of the following agents: prednisolone, thyroxine, puromycin, actinomycin. Both DNA synthesis and tritiated palmitic acid uptake were greatly reduced by puromycin and actinomycin, which killed the cultures in six days. Prednisolone accelerated uptake of lipid precursor and decelerated DNA synthesis; thyroxine treated explants were not different from controls. Lamellar bodies were observed only in the epithelial cells of cultured explants exposed to prednisolone. Similarly, cultures of fetal lung exhibited maturation with appearance of lamellar bodies only when prednisolone was administered. The depression of cellular division and the stimulation of palmitic acid uptake with coicident appearance of lamellar bodies in cultures exposed to prednisolone supports the hypothesis that control of the differentiation of alveolar epithelium with synthesis of phospholipid surfactant, is a function of the adrenal cortex.


Argentaffin granules in endocrine cells in human duodenal mucosa. An attempt was made to gain ultracytochemical characteristics of the endocrine cells in the human duodenal mucosa. Bodian argyrophil and Fontana-Masson argentaffin reactions were applied to the blocks of the tissue. The ultracytochemistry showed that the secretion granules of all morphological types of the endocrine cells were precipitated with both the argyrophil and argentaffin silver grains in varying intensity. The endocrine cells containing uniformly round secretion granules tended to show a heavy precipitation with the argyrophil silver grains in contrast to a light precipitation with the argentaffin silver grains. The endocrine cells containing pleomorphic secretion granules were heavily precipitated with both the argyrophil and argentaffin silver grains. The findings of the present study suggest that the duodenal endocrine cells represent a single cell type in respect to 5-hydroxytryptamine metabolism, but differ in their metabolism of other polypeptide hormones.


Rapid detection of ringed sideroblasts in erythremic myelosis. When normoblasts from patients with chronic erythremic myelosis were stained with the anthraquinone dye, alizarine red S, a purple perinuclear halo appeared. This halo corresponded in location to siderotic granules stained with a conventional iron stain. The rapidity of the alizarine red S stain makes it potentially useful for the detection of pathological sideroblasts in certain refractory anemias.


Immunohistochemical detection of changes in growth hormone cells in rat pituitaries in protein deficiency. 1. An immunohistochemical method was used to study the effect of a low-protein diet on growth hormone (GH) cells in the pituitaries of developing rats. The deficient diet (80 g protein/kg) was administered during gestation and lactation, or during the time after weaning until 90 d of age, or during both periods. 2. GH-cell changes were much more striking in males than in females. 3. In males, GH-producing cells were usually reduced in size and number in all treatments. The effect was most intense when protein deprivation occurred throughout gestation and sucking, and continued until 90 d of age, but it was also evident in animals given the low-protein diet only after weaning. Recuperation appeared to be almost complete when offspring of deprived dams were fed on a normal diet after weaning. 4. It is concluded that a low-protein diet reduces the amount of GH in the rat pituitary in a way similar to that with a protein-free diet.


On the stimulation of viral DNA polymerase activity by nonionic detergent. Nonionic detergents stimulate purified RNA-directed DNA polymerase (reverse transcriptase) activity from various RNA tumor viruses ranging from avian to primate species. The stimulatory effect of the nonionic detergent is dependent on the type and amount of template-primer. The greatest stimulation is obtained when high concentrations of (dT)12-18-(rA)n or activated salmon sperm DNA are used as template-primers. Little stimulation is obtained with viral 70S RNA or with (dT)12-18- (dA)n. The detergent stimulation appears to be specific for viral reverse transcriptase since this effect is not observed with purified bacterial DNA polymerase or with three known mammalian cellular DNA polymerases. This finding may, therefore, be a useful additional criterion for distinguishing viral reverse transcriptase isolated from cells from other cellular DNA polymerases. Nonionic detergent also has a stabilizing effect on viral DNA polymerase against thermal inactivation. This stabilizing effect is further enhanced by the presence of template-primer.


Nature of cold-reactive antibodies to lymphocyte surface determinants in systemic lupus erythematosus. Antilymphocyte antibodies in serum from patients with systemic lupus erythematosus (SLE), as detected by microcytotoxicity and indirect immunofluorescence, were predominantly cold reactive and of the IgM class. These IgM antibodies were most active at 4 degrees C. IgG antibodies were infrequent, and were only minimally lymphocytotoxic. Most sera were cytotoxic for autologous lymphocytes and were equally reactive with normal and SLE lymphocytes, as well as with B- and T-cell preparations. Separate T- and B-cell specificities, which appeared not to be related to HL-A determinants, were identified by differential absorption experiments. The functional significance of these antilymphocyte antibodies is discussed.


Hormonal regulation of the hepatic messenger RNA levels for alpha2u globulin. The messenger RNA rat alpha2u globulin has been identified and quantitated in a cell-free translational system derived from Krebs II ascites cells. Hepatic tissue of the mature male rats which normally produce alpha2u globulin was also found to contain a high level of alpha2u mRNA. Approximately 1.6 per cent of all poly(A) containing RNA of the adult male rat liver could be accounted for alpha2u messenger activity. Female rats do not produce alpha2u globulin and no alpha2u mRNA activity could be detected in the poly(A) containing RNA fraction obtained from the livers of these animals. However, androgen treatment to spayed female rats was found to induce the parallel appearance to both alpha2u globulin and its corresponding mRNA. Both hypophysectomy and adrenalectomy which are known to reduce the level of alpha2u globulin in the urine of male rats were found also to reduce the hepatic level of alpha2u mRNA. The results indicate that hormonal control of alpha2u globulin synthesis in rat liver is achieved primarily through regulation of its translatable mRNA level and that more than one hormone may participate in this regulation.


Sulfhydryl-dependent thermal aggregation of human gamma globulin: augmentation by hyaluronic acid. Hyaluronic acid (4 mg/ml) augmented elevenfold the copper-catalyzed (7 muM) thermal (63 degrees C, 2 hours) aggregation of human gamma globulin (2 mg/ml) in 0.075 M phosphate buffer, pH 7.4. Almost no augmentation of aggregation occurred with hyaluronidase-treated hyaluronate. Hyaluronate-augmented copper-catalyzed thermal aggregation was inhibited by L-histidine, gold thiomalate, N-ethylmaleimide, p-chloromercuribenzoic acid, and ethylenediaminetetraacetic acid. Together with previous reports of a decreased blood histidine concentration in rheumatoid arthritis, these studies provide a possible explanation for the affinity of this disease for joints.


Effects of extracts of rat tissues on the action of bleomycin. The cell sap (105 000 times g supernatant) of various tissues of rats caused DNA degradation in the presence of bleomycin. The activity was fractionated into two peaks by column chromatography on Sephadex G-25. The activity in Peak A (excluded fraction) appeared to be due to some proteinaceous entity, while that recovered in Peak B (retarded fraction), constituting about 90% of the total activity, seemed to be due to ascorbic acid, judging by results of further gel filtration and the effect of treatment with ascorbate oxidase. Incubation of bleomycin with Peak A or B caused loss of the ability of the antibiotic to degrade DNA. It is proposed that the action of bleomycin on DNA, and its inactivation by tissue extracts, depend, at least in part, on the presence of ascorbic acid.


Neutrophil alkaline phosphatase: comparison of enzymes from normal subjects and patients with polycythemia vera and chronic myelogenous leukemia. To determine whether decreased alkaline phosphatase activity in the granules from neutrophils of patients with chronic myelogenous leukemia (CML) was due to an absence of enzyme or the production of defective enzyme, we compared the immunologic properties of granule alkaline phosphatase derived from patients with CML with that of normal subjects and patients with polycythemia vera (PRV). Antisera prepared in rabbits against granule alkaline phosphatase purified from the neutrophils of a patient with PRV produced a single precipitin line of antigenic identity when reacted with extracts of normal, PRV, and CML neutrophil granules. A histochemical stain for alkaline phosphatase activity (alpha-naphthyl acid phosphate coupled with Fast Blue RR) specifically stained the precipitin line. A variety of quantitative precipitin techniques failed to produce satisfactory precipitation of alkaline phosphatase activity. Comparative analyses were therefore performed by affinity chromatography using goat antirabbit-gammaglobulin linked to Sepharose 4B to adsorb alkaline phosphatase complexed with rabbit gamma globulin. With this method, 100% of CML, normal, and PRV alkaline phosphatase could be adsorbed. Using limiting concentrations of antibody, a proportionally smaller fraction of enzyme activity was absorbed as the concentration of PRV alkaline phosphatase or normal alkaline phosphatase was increased. Extracts of CML granules containing comparable amounts of protein but 200-fold less alkaline phosphatase activity per milligram did not specifically reduce adsorption. Thus, in CML, we found no evidence that the granulocytes contained a large amount of antigenically normal but enzymatically defective alkaline phosphatase. Examination of electron micrographs revealed no significant differences in the number or distribution of granules in the granulocytes of normal subjects or patients with PRV or CML. This suggests that the low level of neutrophil alkaline phosphatase in CML granulocytes is the result of decreased enzyme content and not a consequence of synthesis of catalytically defective enzyme.


Morphologic and functional characteristics of bone marrow macrophages from imferon-treated mice. The iron storage macrophage has been isolated from the marrow of Imferon-treated mice and studied in vitro by morphologic, histochemical, and functional tests and isotope labeling techniques. These macrophages on stained preparations are large, many times binucleate cells (up to 150 mu), and show Prussian blue reactivity. In Epon-embedded, stained thick sections they contain elongated narrow basophilic inclusions. These macrophages are actively phagocytic and pinocytic; histochemical studies show that these cells are rich in acid phosphatase, nonspecific esterase, and PAS diastase-resistant activity. Iron storage macrophages do not incorporate the 3H-thymidine. The electron microscopic appearance of this macrophage shows that the cell has ferritin free in the cytoplasm and several types of cytoplasmic granules: those with large quantities of electron-dense ferritin and/or hemosiderin (type A), elongated granules (type B) with moderately electron dense homogeneous matrix and some ferritin at the periphery, and granules with heterogeneous content (type C). The above findings demonstrate that the iron storage cell is a mature macrophage which contains hydrolases, ferritin, and a unique population of cytoplasmic granules which are lysosomal in nature. There is some evidence to suggest that the unusual lysosome (type B granule) occurs after macrophages have ingested erythrocytes.


Cultivation of leukemic human bone marrow cells in diffusion chambers implanted into normal and irradiated mice. In order to elucidate the question of whether the maturation defect in vivo in acute leukemia is due to environmental or cellular factors, we have cultured human leukemic cells in a nonleukemic milieu, i.e., diffusion chambers implanted into the abdominal cavity of normal and irradiated mice. For each harvest, the cell count was measured and differential counts and the number of peroxidase-positive cells determined. The cell number decreased with time, without significant difference between culture in irradiated (500 rads) and normal mice. The blast cells succeeded only in developing distorted promyelocytes and myelocytes. There was a general pattern of increase in the number of peroxidase-positive cells. The study supports the concept that acute myeloid leukemia (AML) is a disturbance of cellular maturation due to cellular rather than environmental defects.


Nature of the longitudinal differential staining of metaphase chromosomes. Patterns of differential longitudinal spiralization of chromosomes induced by the addition of 5-bromodexyuridine (200 mug/ml) for 5-7 h before fixation of the cells were compared with the differential staining of normally spiralized chromosomes with giemsa stain in chromosomes of lymphocytes in human blood cultures. The patterns of differentiation of both types were found to coincide for all chromosomes of the normal karyotype; areas of incomplete spiralization corresponded to areas staining intensively with giemsa stain. The significance of these results in connection with the mechanisms of differential staining of metaphasechromosomes is discussed.