Myelination inhibiting and neuroelectric blocking factors in experimental allergic encephalomyelitis. Sensitization of Lewis rats with whole central nervous system tissue or with purified myelin induced both experimental allergic encephalomyelitis (EAE) and a serum factor which inhibited myelin formation in vitro. Sensitization with the encephalitogenic factor, myelin basic protein, induced EAE, but not the myelination inhibition factor. Sensitization with cerebroside induced neither EAE nor myelination inhibition factor. The serums from control animals without EAE as well as from animals sensitized with all of the above antigens blocked evoked electrical responses in vitro.


Golgi complex--endoplasmic reticulum transition region has rings of beads. The smooth surface of the rough endoplasmic reticulum that makes the forming face of the Golgi complex has beadlike structures arranged in rings at the base of transition vesicles. The beads can only be seen easily after staining in bismuth salts. They are 10 to 12 nanometers in diameter and occur in a variety of cell types and organisms.


Isolation of type C virions from a normal human fibroblast strain. Type C virions were spontaneously released from cultures of a diploid human cell strain. The varions have properties of known type C RNA tumor viruses and share antigenic determinants with the major interspecies-specific antigen (p30) of simian sarcoma virus. Antiserum to reverse transcriptase of gibbon ape leukemia virus inhibits the reverse transcriptase of the putative human virions and that of simian sarcoma virus, but has no effect on the corresponding enzymes of avian or murine RNA tumor viruses.


Hippocampo-hypothalamic connections: origin in subicular cortex, not ammon's horn. An autoradiographic study of the subcortical projections of the rat hippocampal formation shows that the efferent fibers of the hippocampus proper (fields CA1-4 OF Ammon's horn) do not project to the hypothalamus but are confined to the precommissural fornix, ending primarily in the septum. The fibers that are distributed by way of the fornix system to the hypothalamus (principally the arcuate-ventromedial region and the mammillary nuclei) and the anterior thalamus arise from the subicular region of the cerebral cortex (that is, the subiculum, presubiculum, and parasubiculum).


Pseudomonas pseudomallei: II. Laboratory and experimental studies in animals. The cultural, biochemical characteristics and antibiotic sensitivity of strains of Pseudomonas pseudomallei isolated from four cases of melioidosis admitted to Ramathibodi Hospital are described. The organisms were gram-negative bacilli often with bipolar staining. The colonies were wrinkled when incubated for long periods. The characteristic non-specific uptake of dye from media into the colonies and their musty or earthy odour rendered them easily distinguishable from other organisms. All strains were sensitive to chloramphenicol and all but one were sensitive to tetracycline. All strains were resistant to colimycin and gentamicin. The pathogenicity of the strain isolated from a fatal case of peritonitis was studied in guinea pigs. The findings showed that following a large inoculation intraperitoneally, the animal developed acute septicaemia and died shortly afterwards. Only a few micro-abscesses were found on the surface of the liver. Chronic infection of longer duration occurred when a small number of organisms were introduced through a cutaneous abrasion. The lesions included pneumonitis and multiple abscesses of various organs including subcutaneous tissue, liver, spleen and mediastinum.


Antigenicity of sub-cellular components of Entamoeba histolytica. Antigencity of the sub-cellular components of axenic Entamoeba histolytica trophozoites was studied. The cells were disrupted by means of a glass-teflon homogeniser and sub-cellular components were prepared by stepwise differential centrifugation. Four fractions were obtained, namely the 350 g, 6500 g, and 100,500 g fractions and the cell sap. Components of the sedimented fractions were examined by phase contrast and electron microscopy. The antigenicity of each fraction was studied by two different methods:-(1) By extraction with 0.5% sodium deoxycholate followed by testing against the reference sera; (2) By demonstration of the loss of immunological activity of the reference sera after absorption with fractionated components. It was found that all 4 fractions had varying antigenic activities as measured in the indirect hemagglutination (IHA), the complement fixation (CF) and the immunoelectrophoresis (IEP) tests. With the extraction technique, the following results were obtained:- The highest IHA activity was found in the cell sap, whereas this activity in other fractions clustered at lower levels. In the CF test, the activities associated with all 4 fractions were similar. In the IEP test, the highest activity was found in the cell sap and the least activity in the 100,500 g fraction. With the absorption technique, slightly different results were obtained. Whilst in the IHA and the IEP test, the results were in concordance with the extraction technique, the CF activity was slightly different, since it was highest in the cell sap and least in the 100,500 g fraction.


Improvement in the results of treatment of osteogenic sarcoma. Experience with 70 patients having osteogenic sarcomas treated between 1968 and 1973 demonstrated the importance of persistence in the management of the disease of these patients. Resection for pulmonary metastases in one or both lungs, often on more than one occasion, will salvage a number of these patients, thus improving the cure rate. Adriamycin has proved an effective palliative agent and a useful adjuvant to surgical therapy. Although the cure rate of tumors arising distal to the knee is much higher than that of more proximally situated tumors, resection of osteogenic sarcomas of the distal part of the femur does not require removal of the entire femur when care is taken to resect well above the site of the apparent tumor. Some osteogenic sarcomas are sensitive to adequate dosages of radiation therapy, but the exact role of this method is yet to be determined. No definite beneficial result could be attributed to any of a variety of modalities of immune stimulation therapy.


Size changes in single muscle fibers during fixation and embedding. During fixation of single muscles fibers with glutaraldehyde, the volume of the fiber shrinks 20%, recovers in rinse and osmium tetroxide to near normal volume and shrinks 20% again when staining with uranyl acetate. This suggest that osmotic properties of membranes may not have been completely lost during fixation, post-fixation and en bloc staining. Dehydration in ethanol and propylene oxide produces a further 10% shrinkage in volume. Infiltration and embedding with Epon causes an additional 15% change in volume. This gives a total shrinkage in volume of 45% which is nearly twice that of the apparent shrinkage in the volume of the myosin lattice as determined by electron microscopy.


Differentiation of biliary atresia from neonatal hepatitis by routine clinical examinations. After analysis of 33 cases of neonatal hepatitis and 90 cases of biliary atresia, a score test from history, physical findings and daily laboratory examinations was studied for the differential diagnosis of these two diseases. The biliary atresia takes plus score, whereas the neonatal hepatitis takes minus score. The score test is of great value for the differential diagnosis between these two diseases.


An attempt to determine surgical indication for biliary atresia by laboratory examination. To determine the surgical indication of biliary atresia, we analyzed the history, physical findings and laboratory examinations of 90 cases of biliary atresia operated upon at the Second Department of Surgery, Tohoku University Hospital during the past 10 years, from 1963 through 1972 inclusive. A score table from the data of these examinations was made for the determination of surgical indication. According to this score table, all of the patients who showed good excretion of bile took plus score, whereas a half of the patients who showed poor or no excretion of bile took minus score. Although the remaining patients who showed poor excretion of bile took plus score, these cases had some elements for good excretion of bile. From these results, the patients who takes plus score is considered to be a good indication for operation. The score test is of great value for determination of the surgical indication of biliary atresia.


Effects of additives on alum hematoxylin staining solutions. All additives tested (ethyl alcohol, glycerine, chloral hydrate, ethylene and propylene glycol, and citric, malonic and maleic acids) in varying degrees limited the conversion of hematein to insoluble compounds. Peak absorbances increased slightly in hematoxylin solutions containing citric, malonic and maleic acids, but decreased with other additives, and in controls. After four months storage the absorbance in all solutions increased about 50%, acidity increased and staining effectiveness increased.


Hematoxylin substitutes: fluorone black and methyl fluorone black (9-phenyl- and 9-methyl-2,3,7-trihydroxy-6-fluorone) as metachrome iron alum mordant dyes. The phenyl and methyl trihydroxyfluorones, hitherto used histologically only in the rather difficult and unreliable Turchini technics for discriminating deoxyribonucleic from ribonucleic acid, find a new use as iron mordant metachrome dyes which act as nuclear stains. Nuclear staining is unaffected by acid extraction of nucleic acids, as with hematoxylin lakes. The two dyes, named by Liebermann and Lindenbaum 9-phenyl-2, 3, 7-trihydroxy-6-fluorone, have also acquired (illustrating with the phenyl homolog) longer chemical names of the form 2,6,7-trihydroxy-9-phenylisoxanthene-3-one (Eastman). Aldrich and Pfalz-Bauer adhere to the Liebermann-Lindenbaum nomenclature. The trivial name fluorone black is proposed for the phenyl homolog and methyl fluorone black for the methyl homolog. The iron lake of fluorone black appears to be a useful substitute for iron hematoxylin, methyl flurone black less useful. Neither dye has the diverse capability of hematoxylin.


A rapid plastic embedding technique for preparation of three-micron thick sections of decalcified hard tissue. A 24 hour start-to-finish method is described for the preparation of three-micron-thick sections of decalcified hard tissues. Following acetone dehydration, the tissue to be embedded is infiltrated under vacuum  with a series of graded clearing solutions which approach the content of the final methyl methacrylate mixture. After overnight in a 35 C oven, the plastic is polymerized by four hours heating at 42 C. Three-micron-thick sections are then easily prepared by using a Jung microtome for high resolution histologic or detailed autoradiographic procedures.


Tannic acid-iron alum reactions: stain of choice for macroscopic sections of brain to be embedded in plastic. As a macroscopic stain for gross brain sections to be embedded in plastic, tannic acid-iron alum is superior to the generally recommended LeMasurier's variation of the Berlin blue technique because of its greater permanency in plastic. However, as originally adopted for use with brain tissue by Mulligan, the intense black staining of gray matter is too dark for plastic embedded specimens. A modification of this method designed to overcome this difficulty is described. Staining procedure: Wash formalin-fixed brain slices overnight in running water. Wash in distilled water, 2 changes, 30 minutes each. Place slices individually in Mulligan's solution at a temperature of 60-65 C for 4 minutes. Rinse in ice water for 10 seconds. Mordant in 0.4% tannic acid in distilled water for 1 minute. Wash in running tap water for 1 minute. Develop in 0.08% ferric ammonium sulfate in distilled water until gray matter is light gray, about 10-15 seconds. Wash in lukewarm running water for 1 hour, then gently hand-rub whitish film from myelinated surfaces. Store briefly in 3% formalin or 25% glycerine if necessary depending on plastic embedding procedure to be followed.


An overview of botanical clearing technique. Clearing techniques are outlined with reference to their action on the chemical constituents of plant tissue. The most general technique would include pretreatment with solvents, dissolution of protoplasm, dissolution of other substances, bleaching, infiltration with a dense fluid, and staining. Extensive chemical changes go on during these steps and may prevent satisfactory clearing, an important example being the discoloration of phenolic compounds. Rational design of clearing methods for the chemically distinct cell types and tissue seems a likely future development.


A device for the complete elimination of static electricity in paraffin sectioning. In paraffin sectioning static electricity is often induced by friction between the knife and the block. The section can be discharged by ionizing the air in the vicinity of the paraffin. This can be achieved by mounting a strip containing radioactive polonium near the microtome knife. This approach implies a certain radiological risk. An alternative solution is to apply a high electric field close to the knife. The latter method may offer hazards related to the high tension and production of ozone. In comparing the risks and benefits of the two methods we have concluded that the application of a high electric field offers less risks to people in the working area than the installation of a radioactive source. The costs of the two methods are of the same order.


Stains for A, B, and D cells in fetal rat islets. Ten techniques often used for identification of A, B, and D cells in adult islets of Langerhans were applied to fetal rat pancreas. Modifications were tried with many of these techniques. Two indole methods (xanthydrol and postocoupled benxylidene reactions) and a cryostat technique using o-phthaladehyde failed to stain fetal islets. Phosphotungstic acid hematoxylin and lead hematoxylin lightly stained fetal A cell granules in Helly's fixed tissue. The Grimelius silver nitrate technique stains adult rat A cells but failed to stain fetal cells. A modification of this technique stained fetal A cells and a possible 4th cell type. The specificity of this method was confirmed by restaining stained cells with a fluorescent antibody technique and with pseudoisocyanin. B cells, as previously reported, were readily stained by the aldehyde fuchsin technique. Fetal D cells were not stained by the Hellerstrom-Hellman alcoholic silver nitrate method, nor did they display pseudoisocyanin metachromasia after acid hydrolysis; they did fluoresce brightly with this technique when viewed with UV light. It was thus possible to distinguish the three usual cell types, plus a possible fourth type, in the fetal rat pancreas.


Reprecipitation during the preparation of demineralized sections. I. Histochemical and X-ray diffraction observations. The histological features of reprecipitation artefacts occasionally present in demineralized sections of teeth and bone are described. X-ray diffraction analysis indicates that they are crystalline and composed of secondary calcium phosphates.


Reprecipitation during the preparation of demineralized sections. II. Experimental observations. Teeth were incompletely demineralized by immersion in unchanged 10% formic acid for 7 days. Reprecipitation deposits of secondary calcium phosphate were present in the dentin and soft tissues of the dental pulp and, if the final pH was 3 or greater, in the remnants of the periodontal ligament. The deposits in the dentin appeared to be intratubular. Deminieralized sections of teeth suspended in supersaturated solutions of brushite contained similar deposits in the soft tissues. It is suggested that reprecipitation of secondary calcium phosphates is a frequent intermediate stage during demineralization with formic acid.


The Feulgen reaction after glutaraldehyde fixation. A technique is described for performing the Feulgen reaction for DNA on cells and tissues fixed in glutaraldehyde. Blockade free aldehydes by reducing them with fresh 0.5% NaBH4 in 1% NaH2PO4 for 1 hr at room temperature, then rinse in water. Follow by a Feulgen reaction (hydrolysis at room temperature in 6 N HCl for 20 min, Schiff's reagent for 60 min.). Controls assure the completeness and irreversibility of the borohydride blockade. Cytophotometry shows that the DNA content per nucleus is unaffected by the blockade procedure.


Selective light microscopic demonstration of the specific granulation of the rat atrial myocardium by lead-hematoxylin-tartrazine. MacConaill's lead-hematoxylin as modified by Solcia et al. was found to be a highly selective stain for the specific granulation of atrial cardiocytes in the rat. The specific atrial granules were stained blue-black. Contrast was enhanced by counterstaining in a saturated solution of tartrazine in Cellosolve. The stain is compatible with several fixatives and may be used with paraffin or Epon-embedded material.


[Results of cobalt-60-irradiation of brain tumors]. The periods of survival which can be obtained on patients with a primary tumor of the brain after an unique or post-operative cobalt-60-irradiation is dependent of the histology: while periods of survival of several years have been obtained in case of medulloblastoma and astrocytoma, the irradiation of the multiform glioblastoma represents only a palliative measure with a temporary amelioration for any months.


A pattern in the development of agglutinogenic antigens of cyclically transmitted isolates of Trypanosoma gambiense. Three isolates of T. gambiense, belonging to 2 serologically different substrains of the species, were cyclically transmitted to rabbits and monkeys by G. morsitans or G. tachinoides. Sera collected from the infected animals were tested for agglutinating antibodies to 17 different serotype antigens prepared from rodent-adapted isolates of T. gambiense. The first antibodies detected in sera from all of the animals agglutinated the same serotype antigen L2, indicating that it possibly consisted of trypanosomes bearing a basic antigen of T. gambiense. In 2 animals infected with 1 isolate the development of antibody to the common serotype L2 was accompanied by the development of antibody to a second serotype U1. Comparisons of the patterns in which antibodies were produced to other serotype antigens in different animals indicated that there were many similarities in the sequences in which they developed in different rabbits infected with (a) any 1 isolate (b) 2 different isolates of 1 substrain and (c) isolates of 2 serologically different substrains. There were also similarities in the sequences in which the antigens developed in rabbits and monkeys infected with an isolate by the same fly or by different tsetse flies. The ordered sequence in which these 3 cyclically transmitted isolates of T. gambiense, of significantly varied origin and laboratory history, produced variant antigens in hosts of different species provides further evidence to support previous suggestions that antigenic variation in trypanosomes is an adaptive process, rather than one involving selection of mutants.


Relation between malaria serum gamma-globulin concentration and the D-xylose absorption test. D-xylose excretion after a 25 g. oral load was determined in 15 African subjects suffering from malaria. Nine of them were re-investigated after the malaria had been or was being treated. Malarial parasitaemia and pyrexia did not significantly influence results of the D-xylose test. However, in the untreated subjects, there was a significant inverse correlation between the weight of D-xylose excreted and the serum gamma-globulin concentrations. The observation may have a nutritional importance.


[Localization of the fluorescent portion of the Y-chromosome in interphase nulcei of human embryonic kidney cell cultures]. The localization and frequency of accurrence of Y-chromosome fluorescent bodies in interphase nuclei in the cellular culture of 4-months male and female human embryos, were studied with fluorochromine method with quinacrine dehydrochloride. In cellular cultures of male embryonic kidney the frequency of Y-chromatin was about 50%. The Y-chromosome structural connection with nuclear membrane and nucleolus was demonstrated.


A systematic study of testosterone metabolism in benign prostatic hypertrophy (BPH): in vitro results. An in vitro system for testing steroids which might be effective in treating benign prostatic hypertrophy (BPH) has been developed based upon the transformation of H-3-testosterone into the 5-alpha-reduction products dihydrotestosterone and 3-alpha-androstanediol. In scrutinizing the influence of the amount of BPH-tissue, time, and pH, 300 mg of tissue incubated for 2 h at the physiological pH of 7.4 were used in the standard experiment. -The H-3-testosterone concentration was varied from 0.17-100 times 10-8 M. Plotting the resulting 5-alpha-reduction products as a function of testosterone concentration a hyperbolic pattern of enzyme kinetics ensued. Performing a double reciprocal plot of 4 experiments with double determination of each value regression lines could be computed. Those two regression lines most different in their slopes were considered "normal" limits. The rate of H-3-testosterone metabolism could not be enhanced after the endogenous testosterone content within the prostate glands had been used up by means of a preincubation. Scrutinizing the effect of heparin, a weak non-specific enzyme inhibitor, no suppression of the appearance of 5-alpha-reduction products was found. Damaging the BPH-cells, however, by repetitive freezing and thawing lead to an almost complete inhibition of H-3-testosterone turnover.


Sensitivity studies in bacteriologic diagnosis of urinary tuberculosis. The identification of Mycobacterium tuberculosis organisms is insensitive. This in vitro investigation compares the sensitivity of Ziehl-Neelsen and fluorochrome staining techniques after two and ten hours of incubation with standard culture techniques. The culture methods were persistently more sensitive than the stain techniques. Both stain techniques were of equal sensitivity. Ten hours incubation reduces the sensitivity of the staining techniques but does not alter the sensitivity of the culture methods.


Urinary obstruction secondary to prostatic involvement from postoperative extension of carcinoma of rectosigmoid. Two cases of urinary obstruction secondary to neoplastic involvement of the prostate from abdominal-perineal resection for carcinoma of the rectosigmoid are discussed. A review of the literature and the management of this infrequent entity is described.


Use of a Giemsa stain to detect changes in acrosomes of frozen ram spermatozoa. Acrosomal structures of ram spermatozoa were prominently stained when air dried smears of diluted semen were fixed for 15 minutes in buffered formal saline and stained for 90 minutes in a 6 per cent (v/v) buffered solution of Giemsa stain. Progressive disruption of the acrosomes was demonstrated during chilling and deep-freezing of the spermatozoa, and the degree of damage was systematically scored. A rapid and repeatable estimate of the state of the acrosomes in a sample could be made from the mean score of 20 spermatozoa examined per slide.


Demonstration of the Gerbich antigenic determinant on neutrophil leukocytes. Leukocyte-platelet concentrates prepared from blood of Gerbich (Ge) positive donors possess the capability of absorbing anti-Ge. Leukocytes from Ge-negative donors do not absorb the antibody, thus confirming the Ge antigen specificity. Tests with separated leukocyte types suggest that the Ge antigen is present on neutrophils and possibly monocytes, but absent from lymphocytes and platelets.


Cancer of the lung and its response to non-surgical treatment. From July 1969 to December 1972 a clinical trial was carried out to determine the effects of radiotherapy and chemotherapy individually or in combination on lung cancer. During the first three years of the study 53 of 234 patients underwent curative resection. III of the remaining patients were inoperable and were suitable for inclusion in the study. The patients were randomly assigned to four groups: 1) observation only, 2) chemotherapy (Hydroxyurea) only, 3) radiotherapy only, or 4) combination chemotherapy and radiotherapy. There were no differences in survival in any of the treated groups.


[Demonstration of cells of myothelial origin in canine mammary tumours by special staining methods (author's transl)]. Three recent staining methods, the TPA-, TPL-, and TPT-method, were used to demonstrate cells of myothelial origin in mammary gland tumours in bitches and were compared with older techniques. The newer methods proved more suitable for demonstration of myofibrils in myothelial cells. With these techniques it is possible to reveal myofibril containing cells in adenomatous, papillary, myomatour, and myxoid tumour regions but not in chondroid parts of mixed tumours. Two of the tumours examined were classified as malignant myotheliomas because of their staining qualities, structure, ultrastructural appearance and signs of malignancy.


[Inhibition of viral reverse transcriptase and leukemogenesis by modified nucleic acids (author's transl)]. Inhibition of DNA polymerase from oncorna viruses by a new class of macromolecular inhibitors is reported. The macromolecule, designated as mercaptopolycytidylic acid (MPC), is a chemically modified polycytidylic acid containing 5-SH cytidylic bases in the polymerase. Partially thiolated polycytidylic acids (MPC I-III, containing 1.7%, 3.5%, and 8.6% 5-mercaptocytidylate units, respectively) inhibited the DNA-polymerase of Friend leukemia virus (FVL) in the endogenic reaction as well as in the presence of poly rA-(dT)14 or poly (dA-dT) templates; the inhibitory activities were directly related to the percent of tholation. In a bacterial DNA polymerase (E coli-K12 with denatured calf thymus DNA as template) MPCI-III showed no activity. Biological experiments showed that MPC III inhibits the leukemogenic potential of cell-free spleen extracts from FVL-infected mice to about 60%, measured on the basis of spleen weight. The enzymatic and animal experiments have led us to carry out preliminary clinical trials in some cases of Children leukemia. These cases, resistent to the known therapeutic regimes (combination chemotherapy), responded well when treated with MPC along, or in combination with poly I. The experiments indicate that the development of modified polynucleotids with structural similarities to functional templates may be of potential use in the future chemotherapy of leukemia.


[Transposition of great arteries. Results of treatment 1967-1973]. Since Rashkind's Ballon Atrio Septostomy (BAS) was introduced to this Department in Jan. 1967 93 infants with d-TGA were investigated and treated. Of these 70 were included in the study to be presented. Because of major additional anatomic defects, such as valvular atresia, underdevelopment of a ventricle or DORV 23 were excluded. - For Hamburg and the surrounding area with a population of 3-4 million we expected 20 to 30 cases of d-TGA per annum, but only 70 were seen during the 6 1/2 years period of observation. Of these, 64 were palliated by BAS. - Of 70 patients 41 were alive at the age of 6 months, while 29 died within the first 6 month of life. At present 30 are alive, 14 after total correction (Mustard); all 30 were treated with procedures such as BAS, palliative surgery or corrective surgery. - Of 70 patients 37 had simple TGA and 33 combined TGA (VSD, PDA, PS, Coarct.). The life: death ratios of these two groups are identical. - The number of palliative procedures is by far smaller in simple TGA (3) than in combined TGA (23). - The 18 Mustard operations were performed in children between 18 months and 3 1/2 years of age. Since many patients die between the age of 6 and 18 months earlier total correction is imperative. Dysrhythmias and patch shrinkage has occurred twice in 18 Mustard operations. Four patients died within 30 hours following surgery. - Many infants (29) died of simple causes such as respiratory or gastro-intestinal infections during the first 6 months after the initial BAS. A close follow up with frequent x-ray examinations and determinations of hematocrit is recommended in order to increase the rate of survival to the age when total correction is possible.


[Vital fluorescent staining of microorganisms by 3',6'-diacetyl-fluoresceine for determination of their metabolic activity (author's transl)]. By fluorescent microscopy and spectroscopy studies involving micro-organisms which were either viable or devitalised by heat sterilisation or gamma irradiation and could not be cultured any more, fluorochrome binding with 3' , 6'-diacetyl fluorescein was shown to be linked with the viability of a cell and a function of its actual metabolic state. The incorporation of diacetyl fluorescein into cells, its storage and hydrolysis to fluorescein mean active processes taking place at high speed. Viable cells are capable of storing fluorescein intracellularly, bound to structural elements. If the storage capacity is surpassed, it will be eliminated from the cell. The mechanism of this process is discussed. Devitalised cells are not capable of active uptake of fluorescein nor of its storage and accelerated hydrolysis. Beyond this, they are incapable of fluorescein binding to structural elements. There will be only a minor homogenous staining of such cells by fluorescein. An express method based upon the results is providing information on viability, actual metabolic state, morphology, and motility of micro-organisms within a few minutes and without affecting onward culturing.


Antigenic, antibiotic and phage receptor activity of proteins from Sh. boydii 15. The analyzed strain of Sh. boydii 15 induced antibodies against the somatic antigen conjugate, as in most Shigella. Except for the proteinic carrier of the somatic antigen, only a few of the homologous proteinic determinants induced corresponding antibodies; these were illustrated with the help of a serum to proteins from Sh. sonnei. The proteins of Sh. boydii 15 displayed a wide range of antibiotic activity against Shigella, E. coli and Salmonella. Another aspect concerning the biological activities of the proteins isolated from Sh. boydii 15 was its temperate phage receptor activity. This activity was found as well in the free proteins as in the proteinic moiety of the somatic antigen.


[The differenziation of swine-mycoplasma by acrylamide-gel-electrophoresis (author's transl)]. The strains of swine mycoplasma (Tab. 1) were grown aerobically in Whittlestone's Medium containing swine serum for 4-5 days at 37 degrees C. After centrifugation and washing they were freeze-dried and extracted with a phenol-acetic acid-water solution (4:2:1) (1 ml/5 mg mycoplasma dru weight) at 4 degrees C over 48 h. After centrifugation 2 volumes of the supernatant were mixed with 1 volume of a 40% sucrose solution in 35% acetic acid. 0,15 or 0,2 ml of which were added to the acrylamide-gel (2 ml in a tube 0,5 times 10 cm) containing 5 M urea and 35% acetic acid. This solution was overlayed with 0,1 ml 75% acetic acid and tubes then filled with 10% acetic acid. The solution in both electrode chambers was 10% acetic acid, too. During the first 5 min it was separated with 2 mA/tube, then 3 or 5 h with 4 mA/tube. The protein bands were stained with amido black 10B. For control steril culture medium was investigated, too. Various preparations of freeze-dried M. hyopneumoniae gave identical protein patterns. Nearly identical were the patterns of protein bands from M. suipneumoniae and M. hyopneumoniae; that means identity of the strains or very close relationship (Figs. 1 and 2). This is in agreement with other authors who investigated both strains with serological methods. M. suipneumoniae and M. hyopneumoniae were found to be different as well from M. hyorhinis and M. sp. E9 from M. granularum (Figs. 1 and 2). Between M. hyorhinis and M. sp. E9 less relationship was noted. All these results were in agreement with investigations performed with the aid of Latex agglutination.


[Investigations of some metachrome-yellow-preparations as an basic ingredient for metachromgelb-wasserblau-laktose-agar (Gassners medium) (author's transl)]. Experiments with 5 commercial- and 4 testpreparations of Metachrome Yellow have been conducted. Results of this investigations show that the value of Gassners Medium is depending on the quality of the inhibitory substance. The microbiologically active substance (inhibition of grampositive bacteria and prevention of swarming of Proteus) was chemically identified as "Beizengelb GT Color Index 14025" correlating with CI Mordant Yellow I. Test sample II of CHROMA-GESELL-SCHAFT, STUTTGART is recommended as the best "Metachrom-Yellow for preparation of Gassners-Medium Now. Presumable this medium was modified repeatedly after its introduction in bacteriology in 1918 by Gassner. This can be an explanation for the different evaluations of Gassners medium and also for the numerous experiments which have been conducted to modify the medium. Indentity control of chemicals used in microbiology is done by thin layer and paper chromatography. This control should be done in cooperation with chemists more frequently than before.


Heterologous protections in experimental salmonellosis. Immunizations of mice with proteins from S. paratyphi B protected the animals against infection with a concentration of S. paratyphi C which killed the controls and against an infection with 50 LD100 of the homologous S. paratyphi B. The sera of the infected mice showed common precipitation lines of proteins from the two species belonging to different serogroups. Consecutive inoculations with S. typhimurium of both groups of vaccinated mice protected the animals against the infection with their natural pathogen. Immunizations with proteins from S. cholerae-suis protected about 70% of the mice infected with S. paratyphi B and with S. paratyphi C; a higher protection was not, however, induced against infection with the homologous strain. Consecutive infections with S. typhi-murium of the mice resulted in total protection of the animals previously inoculated with S. paratyphi B and S. paratyphi C; the group of mice infected with S. cholerae-suis was less protected against the subsequent inoculation with S. typhi-murium (about 50%). In the infections of mice, S. paratyphi B and S. paratyphi C, seem related to S. typhi-murium by common proteins; the proportion of common protective determinants is not yet known.


Mixed leukocyte reaction and hl-a specificity at multiple sclerosis. Compared with the mixed leukocyte reaction (MLR) between cells from healthy individuals, the MLR between cells from two patients with multiple sclerosis (MS) is, on average, reduced in strength. There is a tendency for the most marked reduction of MLR to occur when both or neither of the members in a tested pair carry HL-A7, and when cells from patients in a relatively early stage of the disease (short duration, relapses but no permanent disablement) are studied; especially when cells from these patients are used as reactor clels in the MLR. Repeated studies of the same cell combinations with varying intervals showed that when both members of the pair carry HL-A7,--and therefore have a large chance also to carry LD-7a--the MLR strength is relatively constant. When only one or neither of the members carry HL-A7, the strength of MLR varies considerably. Probably two different mechanisms exist in MLR impairment: similarity of LD genes, and "immunosuppressive" factors which can vary during the course of the disease.


The basement membrane and lobular carcinoma in situ of the breast. The basement membrane (BM) was examined by light microscopy in cases of lobular carcinoma in situ of the breast (LCIS), employing the following stains: haematoxylin-eosin, van Gieson-Hansen, PAS, colloidal-iron-PAS, and reticulin. The BM was non-intact in 12 out of 26 investigated cases of LCIS when reticulin staining was applied, but appeared non-intact in about 70 per cents with the other staining methods. On the other side control areas of the BM were intact in 23 out of 26 cases with reticulin stain, but in only about half of the cases with other stains. It is concluded that reticulin staining is the most suitable in evaluating the BM in LCIS, and that more or less pronounced defects of the BM in LCIS cannot be considered evidence of imminent invasion. Although the overall occurrence of invasive breast carcinoma was significantly increased in 9 cases with epithelial cell protrusion through the BM, this feature should be investigated further before it is allowed to affect therapeutic decisions.


A fluorescence and electron microscopic study of the adrenergic innervation in the vestibular ganglion and sensory areas. The adrenergic innervation in the vestibular ganglion and sensory areas in rabbit and cat was studied with fluorescence and electron microscopic techniques. All adrenergic nerve fibres consisted of postganglionic axons from the ipsilateral superior cervical ganglion. In rabbit, postganglionic nerve fibres passed via the carotic plexus. Probably because of the anatomical difference, the feline internal carotic artery being rudimentary, all sympathetic nerve fibres to the inner ear in cat passed via the tympanic plexus. In the vestibular ganglion there was a uniform distribution of adrenergic nerve fibres and there was no difference in distribution patterns between rabbit and cat. There was a continuous blood vessel innervation and an innervation independent of blood vessles. The adrenergic innervation extended to the sensory areas but did not seem to penetrate the basement membrane and no adrenergic nerve terminals were found in direct contact with sensory or secretory cells.


Demand pacemaker arrhythmias caused by intermittent incomplete electrode fracture. Diagnosis with testing magnet. This report describes how the testing magnet was used to diagnose intermittent and incomplete electrode fracture in two patients with an implanted demand pacemaker. During fixed-rate pacing the interval between two consecutive pacemaker spikes intermittently doubled in length, suggesting that the pulse generator was continuing to fire on time into a transiently disrupted circuit. Attenuated pacemaker spikes occurring at the anticipated time of pacemaker discharge also provided a diagnostic clue. Ventricular electrograms from the defective electrodes registered small false signals.


In-vitro platelet-neutrophil adherence.

In-vitro platelet-neutrophil adherence. The specific adherence of platelets to neutrophils in vitro has been noted with increasing frequency and with increasing interest, but its significance remains obscure. Two cases in which this event occurred are presented, and the literature is reviewed.


Hemolytic anemia caused by auto anti-N.

Hemolytic anemia caused by auto anti-N. The second case of hemolytic anemia caused by auto anti-N, occurring in a 7-year-old boy, is described. The antibody was IgG, as shown by the use of specific anti-human IgG Coombs sera, failure of inactivation by 2-mercaptoethanol, and chromatographic separation on a G-200 Sephadex column.


A technic for bone marrow imprints from postmortem specimens. A simple technic for making bone marrow imprints from postmortem specimens is described. The preparations are highly reproducible, retaining excellent cytologic detail with minimal cell distortion and can be used for evaluation of cellularity.


Hematoxylin substitutes. A study of phenocyanin TC and the use of afterchrome mordanting in histology. A practical, simple synthesis of the obsolete mordant dye, phenocyanin, was devised, proceding from gallocyanin and resorcinol with acid and heat. The dye gave promise of good performance in metachrome iron mixtures, but because of excessive precipitation, the practice of afterchroming was taken from textile dyeing usage, and proved very successful. Of a number of metallic salts tried, Fe II proved to be the best, then Cu II and Fe III. The stain acts as a cationic dye on nucleic acids and other acidic tissue components: acid mucins, cartilage, mast cells, corpora amylacea, etc. The afterchroming process renders the stain much more resistant to various extraction agents and even moderately resistant to acid alcohol. Color values are quite comparable to those obtained with hematoxylin-eosin when an eosin counterstain is used. Nuclei basophilic cytoplasm, Nissl granules, and bacteria color dark blue; cartilage, mast cells, and some acid mucins, deep violet. Staining with the 1% solution was essentially unaltered from neutrality down to 1.2 N HCl, pH 0.68, and dilution of the dye to 0.05% in 1% conc. HCl (pH 1.2) still gave excellent nuclear, RNA, and mast-cell staining. At 0.02% and pH 1.2, nuclear staining was distinctly weakened; mast cell granules were still dark violet.


Unusual ultrastructural features of a leiomyosarcoma of the lung;. The electron microscopic features of a leiomyosarcoma occurring in the lung, a rare site, were studied; Unusual was the fwnding of extremely electron-dense bands that interrupted the longitudinally-arranged microfilaments and dense bodies of many of the tumor cells. These bands superficially resembled anomalous Z-bands seen in nemalin myopathy and aging rat cardiac muscle, but have not been described to occur in smooth muscle tumors. They are thought to represent condensations of dense bodies in degenerating tumor cells.


Unusual ultrastructural features of leiomyosarcoma of the lung. The electron microscopic features of a leiomyosarcoma occurring in the lung, a rare site, were studied. Unusual was the finding of extremely electron-dense bands that interrupted the longitudinally-arranged microfilaments and dense bodies of many of the tumor cells. The bands superficially resembled anomalous Z-bands seen in nemalin myopathy and aging rat cardiac muscle, but have not been described to occur in smooth muscle tumors. They are thought to represent condensations of dense bodies in degenerating tumor cells.


Hydralazine-induced lupus erythematosus-like syndrome. A reversible syndrome resembling systemic lupus erythematosus and induced by hydralazine hydrochloride therapy is a well-recognized phenomenon in adults but does not seem to have been reported in children. A 9-year-old girl had fever, arthralgias, modest joint swelling, splenomegaly, antinuclear antibodies (ANAs), anitbodies against native and denatured DNA, and positive LE cell preparations after nine months of hydralazine hydrochloride therapy, 120 mg/day. Clinical findings returned to normal within four weeks of discontinuing the drug therapy, and serological abnormalities disappeared after 11 months. Like previously reported patients, the child is white and has a slow acetylation phenotype. It is not known whether children receiving hydralazine are as susceptible to this complication as adults. Periodic ANA determinations may be advisable for children receiving hydralazine, especially if they are white and have a slow acetylation phenotype.


Norlestrin-induced pseudopregnancy in the baboon. Effects on blood coagulation and endocrine glands during attempted production of Sheehan's syndrome. Production of Sheehan's syndrome was attempted. As it was not possible to obtain pregnant baboons initially, pseudopregnancy was induced with norlestrin, 2.k mg., for 3 and 4 months, respectively. Sheehan's syndrome was not produced with massive hypovolemic shock but failure to obtain true pregnancy has given serendipitous information on the long-term action of norlestrin on the pituitary and its target organs. Unusual abundance of prolactin acidophils and decrease of growth hormone acidophils was found. Further studies will be necessary to show the possible significance of these findings. As fibrin degradation products did not appear in the serum, we can make no inference about the necessity of intravascular coagulation for the production of Sheehan's phenomenon. Finally one animal became pregnant. Sheehan's phenomenon was not produced in her despite induction of intrapartum hypovolemic shock. Histologic findings in her pituitary resemble those of the pseudopregnant animals.


Intrauterine diagnosis and control of fetal ventricular arrhythmia during labor. A very rare case of a sustained fetal ventricular arrythmia in the form of bigeminy, trigeminy, and quadrigeminy during labor is described. The rhythm distrubance failed to respond to sedatives and narcotics but was successfuly reverted to sinus rhythm following the administration of intravenous propranolol to the mother. The significance and possible mechanism of the arrhythmia is discussed.


Thyroid disease in pregnancy.

Thyroid disease in pregnancy. It is frequently difficult to establish a diagnosis of hyperthyroidism in association with pregnancy. Signs and symptoms suggestive of hyperthyroidism may occur in normal pregnancy. Moreover, tests to rule out hyperthyroidism under these conditions are not readily available. Mild hyperthyroidism is not a hazard to an otherwise normal pregnancy and does not require therapy on the basis of this presumption. Hyperthyroidism of a clinically significant degree is safely treatable by medical means without hazard to the fetus. Hyperthyroidism is an uncommon cause for the failure of pregnancy to proceed to term. Treatment with thyroid based on a presumption of this diagnosis is justified, but such patients should be studied carefully after delivery to establish the true state of thyroid function for the future. Other conditions of thyroid dysfunction, including thyroiditis, thyroid carcinoma, nontoxic goiter, and ovarian struma rarely interfere with an otherwise normal pregnancy.


Caveola--vesicle complexes in the plasmalemma of erythrocytes infected by Plasmodium vivax and P cynomolgi. Unique structures related to Schüffner's dots. Erythrocytes infected with Plasmodium vivax and P cynomolgi, characterized by Schüffner's dots on Giemsa-stained thin films, were studied by electron microscopy and immunocytochemistry. A caveola-vesicle complex, which consisted of a caveola surrounded by vesicles, in an alveolar fashion, formed along the erythrocyte plasmalemma. Horseradish-peroxidase-labeled immunoglobulin from a monkey infected with P vivax bound to the vesicle membrane. Cationized ferritin appeared within the vesicles after incubation with viable parasitized erythrocytes, suggesting that these vesicles were pinocytotic in origin. This caveola-vesicle complex probably corresponds to Schüffner's dots because the alteration is unique to vivax- and ovale-type malarias, and its size and distribution are consistent with Schüffner's dots. Clefts observed within the cytoplasm of infected erythrocytes are present in all malarias and are unlikely candidates for Schüffner's data.


Comparison of two occupational therapy approaches for treating the young cerebral-palsied child. This pilot study compared the results of facilitation and functional treatment approaches on the overall development of 12 cerebral-palsied children aged 1 to 5 years. The Denver Developmental Screening Test and the Bayley Motor Scale were used to evaluate the subjects. Pairs of children with similar developmental ages were identified and randomly assigned to the two treatment programs. Posttests and subsequent statistical analyses indicated that the group of children receiving the facilitation treatment approach showed significant improvement then did the group receiving the functional treatment approach in computed developmental age. Results suggest that comparative and evaluative studies should be continued on these approaches, as well as on other areas of clinical practice, to validate the effectiveness of occupational therapy treatment.


Comparison of straddling and sitting apparatus for the spastic cerebral-palsied child. The sitting posture of a spastic cerebral-palsied child in a standard chair and on a straddling apparatus is discussed in this paper. It is suggested from the literature that a sitting apparatus which encourages hip abduction positioning provides the stability a child needs to engage in bilateral hand skills. In examining the sitting posture of a cerebral-palsied child, it is recommended that straddling apparatus be used.


Preventing the abuse of little children: The Parents' Center Project for the study and prevention of child abuse. Forty-six families with 73 children between the ages of six months and four years were treated in a project developed to preserve the integrity of the family while protecting the child from physical abuse. Improvement in the rate of growth and development was found among the children. There was much less improvement in the domestic functioning of their parents. However, it appeared that the parents would not have kept their children in the project if they had not been in concurrent treatment.


A summer program for underachieving adolescents. This paper discusses a Summer Adolescent Program organized for mentally and physically handicapped teenagers with potential for a moderate level ofindependence, but lacking in self-help, socialization, and mobility skills, and prevocational readiness. The purpose of the seven-week program is to provide a practical, activity-oriented setting where teens can capitalize on existing skills, and learn and practice new skills. Program content includes assembly project, personal care and cooking units, recreation, interaction groups, and parent participation. Evaluations are based on objectives formulated for each adolescent at the beginning of the program and on staff ratings of contacts with each youngster.


Ganglion cells in arterioles of skeletal muscle: role in sympathetic vasodilation. Ganglion cells were found in arterioles of gracilis muscles of dogs 2 wk after complete extrinsic denervation. We tested the possibility that they function in active sympathetic vasodilation (SVD) induced in isolated gracilis muscles by hypothalamic stimulation. To this end various drugs were injected into the gracilis perfusate. (Drugs did not reach the systemic circulation, and exerted their effects within the gracilis itself). C6 had no effect. Submaximal doses of atropine or l-hyoscyamine delayed and slowed SVD; higher doses blocked completely. d-Hyoscyamine did not change SVD or acetylcholine vasodilation, but when administered prior to l-hyoscyamine, threshold for blockade of SVD by the l-isomer increased 10,000-fold. Blockade by l-hyoscyamine of acetylcholine vasodilation was unaltered by d-hyoscyamine. Eserine partly blocked SVD, but enhanced and prolonged acetylcholine vasodilation. The foregoing and certain features of the time course of SVD are interpreted to mean that: a) atropine blocks SVD at muscarinic sites on peripheral ganglion cells; b) transmission at the ganglion cells depends on slow excitatory postsynaptic potentials; c) the final mediator of SVD at the vascular muscle cell is unknown.


Implicit messages concerning menstruation in commercial educational materials prepared for young adolescent girls. Interviews with young girls aged 9 to 18 revealed that commercial educational materials are an important source of information about menstruation. Although these materials are valuable educational supplements, they do not provide a complete, accurate, and realistic description of menarcheal changes and emphasize good hygiene rather than dealing with the young girl's emotional needs and anxieties. The authors stress the need for more research into the physiological, cognitive, and psychological changes during puberty and the obligation of parents, schools, and other social institutions to provide more comprehensive information to the maturing girl about the essential issues of pubertal development.


[Allergic reactions following dextrane infusions (author's transl)]. 14 allergic reactions to dextran 60 and dextran 75 are reported. In 8 patients the peripheral pulse became impalpable; Medical history and clinical examination did not reveal any predisposition. Histamine liberation is to be discussed. Antihistamines did not protect against allergic reactions of colloid plasma expanders. All the patients were treated with prednisolone and fluids. After rapid infusion of Ringer-lactate and plasma protein solution to an amount of 2000-2775 ml adequate circulation was restored.


Composition of antigens of various mycobacterial species detected with a Mycobacterium tuberculosis reference serum. Immunoelectrophoresis with a reference antiserum to Mycobacterium tuberculosis permitted the identification of antigens shared by 12 species of mycobacteria. No antigens unique for M. tuberculosis were found.


Hepatocellular carcinoma and a variant alkaline phosphatase. Four patients with hepatocellular carcinoma had a variant alkaline phosphatase that resembles the placental D-variant but is different from it in electrophoretic mobility, pH optimum, heat stability, and inhibition by phosphate. The appearance of this enzyme has been specific to hepatocellular carcinoma. Its prevalence was about 30%, while that of another marker protein, alpha-fetoprotein was 77%. The occurrence of this enzyme in serum of patients with hepatoma was, accordingly, independent of the serum alpha-fetoprotein concentration, and also independent of the appearance of the Regan or the Nagao isoenzymes and of the serum alkaline phosphatase activity. Patients with the enzyme had a massive type of hepatocellular carcinoma with grade III differentiation by Edmondson's classification. The detection of this enzyme in serum may be of help in confirming the diagnosis of hepatoma.


Substructural analysis of the microtubule and its polymorphic forms. Microtubules are composed of 13 protofilaments which are aligned parallel to the long axis of the tubule, as can be seen in thin sections of material fixed in the presence of tannic acid. Evidence is presented based upon Markham rotations of the isolated flagellar axoneme that the bridges (spoke, nexin, dynein) must connect to certain specified protofilaments. By means of geometric considerations it is concluded that the bridges in a Centrohelida and Raphidiophrys also connect certain specified protofilaments. Two polymorphic forms of microtubules were also examined using tannic acid in the fixative, a 320 A tubule produced in Echinosphaerium by treatment with low temperature, and a vinblastine-induced microtubule crystal. In both forms, the arrangement of the subunits is not in the form of linear protofilaments. The vinblastine crystal in transverse section is made up of hexagonally packed tubules approximately 320 A in diameter. The wall of each tubule in turn is composed of two parallel protofilaments which spiral upwards, giving the appearance of a barber pole. The center-to-center separation of adjacent protofilaments is 180 A. We conclude that the tubulin heterodimer lies parallel to the axis of the protofilaments, exposing the colchicine binding site.


Action of colchicine on axonal flow and pituicytes in the hypothalamopituitary system of the rat. Changes in the hypothalamo-pituitary tract and the pituicytes of the rat were studied after intrathecal and intraperitoneal injections of colchicine. Radioautography with 35S-cysteine demonstrates that intrathecal colchicine prevents the migration of neurosecretory granules from the supraoptic and paraventricular nuclei to the posterior lobe of the pituitary. This results in accumulations of neurosecretory granules and in the formation in the axons of elongated structures resembling neurosecretory products, although they sometimes have a fibrillary content. Neurotubules appear to remain intact in these conditions. The stimulation of the posterior pituitary by dehydration, in particular after injection of the diuretic furosemide, leads to an increased activity of pituicytes. When colchicine is injected at the same time as furosemide, a considerable new formation of centrioles is observed in the pituicytes. These become associated with ciliary vesicles, and form numerous cilia of the 9 + 0 type. An increased number of centrioles is also seen in the endothelial cells of the posterior lobe of the pituitary. These apparently paradoxical results were briefly discussed in relation to the action of colchicine on neurotubules and axonal flow and to the limited data from the literature indicating a stimulation of cilia formation under the action of colchicine and other drugs.


Retinal and pigment epithelial alterations over choroidal malignant melanomas. Although in some cases orange or yellow-orange pigment overlying a malignant choroidal melanoma may represent lipofuscin, a case is presented which showed extensive yellow-orange coloration overlying a malignant choroidal melanoma without any demonstrable lipofuscin on histopathologic and electron microscopic analysis. Our findings suggest that other mechanisms, such as serous fluid or lipid deposition within retinal cystoid spaces or diffuse drusen, can clinically stimulate the yellow-orange pigment associated with lipofuscin.


Museum preparations of the conducting system of the heart. A technique of dissection of the sinuatrial node and of the atrioventricular node and main conducting bundles of the ox heart, without injection, is described. Museum specimens prepared in this way are illustrated.


Supernumerary bisatellited chromosome in a family ascertained through a patient with Sturge-Weber syndrome. The presence of a structurally abnormal extra chromosome in a patient with Sturge-Weber syndrome and several members of her family is described. With routine techniques the abnormal chromosome is slightly submetacentric, of the size of a G group chromosome and shows satellites on both arms. C-banding suggested the presence of 2 centromeric regions rather than one, and to explain this finding, in addition to the segregation of the abnormal chromosome through 3 generations and why only one centromere is visible with the usual cytogenetic technique, an hypothesis is advanced suggesting that it resulted from an unusual type of Robertsonian translocation, in which one of the breacks involved directly the centromere of an acrocentric producing a partially dicentric bisate-lited chromosome. The association of Sturge-Weber syndrome with the chromosome abnormality is thought to be fortuitous and the lack of clinical manifestations of all members of this family with the abnormal chromosome, including one with two extra ones, is explained by the fact that it was almost entirely formed by heterochromatic material. The usefulness of C-banding in the study of this patient is strongly emphasized.


Partial trisomy 10q occurring in a family with a reciprocal translocation t(10;18)(q25;q23). Partial trisomy 10q was observed in an eighteen year old girl with severe mental and physical retardation, microcephaly, a high forehead, microphthalmia, antimongoloid slants, low set ears and severely malformed extremities. A balanced translocation t(10q-;18q+), present in several family members, was identified by fluorescence and thermic denaturation techniques; the break points were 10q25 and 18q23. A comparison made with seven similar cases suggests a common, phenotypical appearance which may be of diagnostic value.


17 long arm isochromosome. A common anomaly in malignat blood disorders. A cytogenetic anomaly consisting in the replacement of a 17 by its long arm isochromosome was identified as the only alteration in the marrow cells of two patients with acute granulocytic leukemia. In one case, the specific nature of the abnormal chromosome was established by newly available techniques. Since its identification in 1965, this structural anomaly, which implies 17 long arm duplication and short arm deletion, has been observed, as a sole or as an associated finding, in the malignant cells of a spectrum of blood disorders, including acute granulocytic leukemias, the blast crisis of chronic myeloid leukemia and lymphoreticular proliferative disorders. Attention is called to this particular rearrangement for its clinical as well as fundamental implications, as its presence in blood forming cells unfailingly hearalds a fast, fatal course of evolution.


[2:12 (p25;q21) translocation classed at first as 2/X]. A balanced translocation t(2;12)(p25;q21) was observed in a girl with mental retardation and several somatic abnormalities. The problems of phenotype-karyotype correlations are discussed.


The total correction of congenital heart disease in infants. If radical correction of congenital heart disease can be performed with a low risk in the first year of life the advantages are obvious. Total correction avoids the latent risks of the underlying lesion and the risks of palliative surgery, and relieves the parents and family of the psychological pressure of a major illness. Palliative surgery, although offering survival, may include the long term problems of the palliative operation itself, and the possibility that cases may be lost to further treatment or follow-up examination after a successful palliative operation. For these reasons, to be desirable, a palliative operation should offer a survival chance that is at least 10% better than the corrective procedure in that patient at that time. That is, any corrective procedure with a mortality rate less than 10% is to be perferred at any age to a palliative operation. An accurate assessment of risk demands a complete diagnostic study, and a knowledge of the natural history of that form of congenital heart disease. The problems of infants with congenital heart disease are not primarily caused by age of size. The main problem is that of natural selection.


Inhibition of dihydrostreptomycin action on Mycobacterium smegmatis by monovalent and divalent cation salts. We have examined and compared the effects of monovalent and divalent cation salts on dihydrostreptomycin (DSM) action against Mycobacterium smegmatis. The Sauton synthetic liquid medium used was supplemented with test salts on the basis of ionic strength (mu). Turbidimetric growth experiments showed that 0.02 M MgSO(4) (mu = 0.08) prevented growth inhibition by 0.1 mug of dihydrostreptomycin per ml, but 0.02 M NaCl (mu = 0.02) did not. However, at molarities equivalent to mu = 0.08, four monovalent cation salts, including NaCl, Na(2)SO(4), NH(4)Cl, and (NH(4))(2)SO(4), all prevented inhibition by dihydrostreptomycin. When magnesium and sodium salts were compared at mu = 0.02, 0.04, and 0.05, two distinct growth protective patterns were seen. These data were indicative of two different mechanisms of dihydrostreptomycin antagnosim by salts; the first being divalent cation and concentration dependent, and the second being nonspecific and ionic strength dependent. Viability studies supported the existence of two mechanisms.


Presence of two virus-like particles in Penicillium citrinum. Two icosahedral virus-like particles (28 and 19 nm in diameter, respectively) have been detected in sporogenic and asporogenic segregants of a strain of Penicillium citrinum. The distribution of the two particles differed among the two segregants.


Intestinal lymphangiectasia. Long-term results with MCT diet. The clinical course of 6 children with primary intestinal lymphangiectuasia who have been treated with low fat medium chain triglyceride-supplemented diets for between 3 and 8 years (4 for longer than 5 years) is described. Though laboratory findings indicate continuing chyle leak, evidence for long-term benefit from dietary treatment is provided by symptomatic relief while on the diet, clinical relapse upon relaxation of t,e regimen, and improvement in growth rates. In most patients the underlying lymphatic defect, and thus the need for dietary treatment, appears to be permanent.


[Formation of elastic fibers around basal cell carcinomas: light microscopical observations (author's transl)]. From 275 specimens of all sorts of basal cell epitheliomas the new formation of elastic fibers was histologically investigated by means of selective elastic tissue stains such as orcein and resorcinfuchsin. In nearly one third of the basal cell epitheliomas newly formed elastic fibers could be observed in the stroma. In places where a new formation of elastic fibers was demonstrated fibroblasts and mast cells (toluidine blue stain) were unusually numerous. Apparently the influence of the growing tumor masses of basal cell epitheliomas leads to a dermo-epidermal interaction in the form of 1. increase of mast cells 2. stimulation of fibroblasts 3. new formation of elastic fibers. The observation suggest that there exists a relationship between the above noted findings. Due to the dermo-epithelial interaction (even in senile skin) in the stroma of basal cell epithelioma fibroblasts can be stimulated to such a degree that they regain the ability to produce elastic fibers-a function which usually is greatly diminished with increasing age. The findings seem to be important from the view point of rejuvenation of aging human skin.


Elemental analysis of honey as an indicator of pollution. Forty-seven elements in honeys produced near highway, industrial, and mining areas. Nineteen samples of honey taken from grocery shelves, near zinc mines, adjacent to an industrial area, and near a major highway, and one sample collected in 1899 were subjected to spark source mass spectrometry to determine the concentration of most of 47 elements in the honey. Certain samples of honey produced by bees in the vicinity of the New York State Thruway appeared to contain elevated levels of certain elements known to be emitted by traffic, such as aluminum, barium, calcuim, copper, magnesium, nickel, palladium, and silicon, but the true source of these elements was uncertain. Honey that had come into contact with metal containers in processing contained the greatest amounts of zinc and tin.


Preliminary immunologic characterization of glycoprotein antigens (CEA) of tumors of gastrointestinal tract in humans. Using the method of double diffusion in agar and precipitation, inhibition test and specific antitumor immune sera, perchloric extracts of carcinomas of the stomach and lower parts of the gastrointestinal tract were found to contain a carcinoembryonic antigen with electrophoretic mobility of gamma-globulins. Antigens from carcinomas of all studied parts of the gastrointestinal tract (stomach, sigmoid, colon and rectum) gave lines of identity in the test of double diffusion in agar. This antigen was not demonstrable in the mucous membranes surrounding the carcinomas of the gastrointestinal tract or in normal organs (gastrointestinal mucosa, liver, kidneys, spleen, plasma), or in perchloric extracts of gastrointestinal tissues of human fetuses.


Infectious bursal disease agent: morphology by negative stain electron microscopy. The virus of infectious bursal disease of chickens was studied by immune electron microscopy. Negatively-stained preparations revealed morphological similarities with both the bluetongue virus group, and the virus of infectious pancreatic necrosis of trout. Results indicated that the small particle found in such preparations is a degradation product of the large particle.


Bleomycin: action on growth of oncogenic RNA viruses and on cell transformation. Bleomycin (BLM) inhibits cell proliferation of noninfected chick embryo fibroblasts by blocking their DNA synthesis selectively. Chick embryo fibroblasts have beentransformed by Schmidt-Ruppin D strain of Rous Sarcoma Virus. Transformation has been determined by a focus assay. Foci formation is strongly reduced by BLM. Virus replication is inhibited by BLM in growing and confluent monolayer cells. This result might be explained by the observation that this drug reduces proliferation of growing and of confluent monolayer cells very sensitively. During the first 24 hours after infection the BLM inhibitory effect is more pronounced than in the case of BLM-application during the period 24--48 hours after infection. This result is explained by published results, showing that cell division is required only for the intitiation of transcription of virus RNA but not for its maintenance. BLM has only little effect on virus growth in transformed cells, because in these cells initiation of transcription of virus RNA has already taken place before the drug was added. From the data obtained it is concluded, that BLM inhibits growth of Rous Sarcoma Virus by blocking cell proliferation.


Circulating immune complexes. Effects on ocular vascular permeability in the rabbit. Sudden transient, alteration in vascular permeability is producible in rabbit eyes by intravenous injection of large quantities of antigen, bovine gamma-globulin (BGG), into immunized animals or by intravenous injection of large amounts of antigen-antibody complexes (BGG-antiBGG) prepared in antigen excess (20 to 25times) in normal rabbits. Change is measured by ocular accumulation of iodinated I125 serum albumin, relative to that in heart blood, in intact eyes and separate anatomical compartments, in aqueous, in the anterior eye including (lens plus vitreous), and in the posterior segment. Primarily affected are vessels in the iridial portion of ciliary processess; edema is the primary finding. The same vessels are affected by intravenous injection of bacterial endotoxin, but compared with endotoxin, altered vascular permeability is short-lived and is not associated with formation of platelet plugs and intravenous fibrin strands.


Pituitary adenoma producing amyloid-like substance. Light and electron microscopical examination of a pituitary adenoma showed that the tumor was composed of prolactin cells that produced amyloid-like material. The findings have been interpreted in favor of the role of rough endoplasmic reticulum in amyloid fibril formation within the adenoma cells. Histochemical and fine structural data indicate that the material produced by this tumor resembles apudamyloid rather than immunoamyloid.


Adrenocortical postirradiation fibrosis. At autopsy, the adrenal zona reticularis showed linear hyaline fibrosis in 19 cases of cancer, after averages of six months and 5,000 roentgens of irradiation to the abdominal, lumbar, or pelvic regions. The associated roentgen dose was significantly greater than when postirradiation adrenal fibrosis was absent. Primary radiation damage to the reticular vascular plexus is suspected.


Perianal Paget disease. Histochemical differentiation utilizing the borohydride-KOH-PAS reaction. Specimens from various types of Paget disease, other tumors, and certain normal tissues were examined with a battery of histochemical techniques, including the sodium borohydride-potassium hydroxide-PAS method that specifically stains certain sialomucins that are found in terminal parts of the ileum and of the colon. These sialomucins were present in normal anal ducts but were not present in transitional or anal-covering epithelium. A case of perianal Paget disease showed strongly positive staining, both in the underlying mucinous adenocarcinoma and in Paget cells of the affected anal and perianal skin. In contrast, stains of other forms of Paget disease were totally negative with this technique, as well as malignant melanoma and Bowen disease. These results support the theory that Paget disease represents epidermal invasion by malignant cells from underlying tumor.


[Experimental study of the correlation between function and morphology of rat thyroid after the application of methylthiouracil and thyroid stimulating hormone]. In an experimental study with 359 male Wistar rats the thyroid function was altered with MTU and TSH. The morphological changes developing due to this treatment were compared with the hormone blood level measured as labelled protein-bound iodine (PBI-127). Furthermore, the content of PBI-131 was determined and the I-131 uptake of the thyroid after application of TSH was measured. It was shown that with a dosis which had no influence on the PBI-127 and radioiodine values, the height of the thyroid epithelium already revealed a significant reaction. By a morphological examination together with the histometrical determination of the height of the thyroid epithelium the hormone-dependent increase in activity may be determined quantitatively.


[Correlation of in vitro testing and therapeutical results in animal transplanted tumors after cytostatic treatment]. The antineoplastic activity of 5 substances was tested in vivo and in vitro on four different tumours (plasmocytoma and melanoma Fortner III of the Syrian golden hamster, the Walker carcinosarkoma 256 and an adenocarcinoma of the rat). The substances involved were 2,3,5-triethyleniminobenzoquinone-(1,4) (triaziquone), actinomycin D, podophyllinic ethylhydrazide (mitopodozide), bleomycin and adriamycin hydrochloride. The effect of the substances in vivo was measured on the size of the tumour, and in vitro on the incorporation of 3-H-thymidine and 3-H-uridine in short-term incubations of tumour-cell suspensions. No correlation was observed between the 3-H-thymidine incorporation in vitro and the response of the tumours in vivo. On the other hand, the 3-H-uridine incorporation in the tumour-cell suspensions in vitro was in good agreement with the results of therapy in the animal experiments. This is compatible with the results of earlier experiments using other substances to investigate the possible correlation between tumour therapy and in vitro tests.


Evaluation of arrhythmias in the late hospital phase of acute myocardial infarction compared to coronary care unit ectopy. To evaluate the prevalence and nature of arrhythmias during the entire three-week period in hospital after myocardial infarction, the results of coronary care unit monitoring (initial 3 to 5 days) were compared with continuous 8-hour portable monitoring during the ambulatory phase (second and third weeks) in 83 consecutive survivors. Arrhythmias were detected in 84.3 per cent (70/83) of patients while in the coronary care unit and in 85.5 per cent (71/83) during hospital stay after the coronary care unit. Ventricular ectopic depolarizations were classified as complicated (multifocal, paired, R on T, or five or more a minute) or uncomplicated. Importantly, the high frequency of complicated ventricular extrasystoles and tachycardia persisted during the entire period in hospital (early 34.9% and late 42.5% of all patients). However, only 16.9 per cent (14/83)had these ventricular arrhythmias during both coronary care unit and ward monitoring. Thus, the absence of complicated ventricular ectopic depolarization and ventricular tachycardia in the coronary care unit did not exclude their subsequent occurrence in the majority of the large number of patients with late hospital complicated ventricular ectopy.


Lipid in the Achilles tendon. A comparative study. The tendo Achillis of man and its homologue from a fairly wide range of exotic animals, most of which had died in captivity from natural causes, have been examined for stainable lipid. Adiposity of the tendon was seen principally in man. Chondroid plaques, containing an abundance of intracellular triglycerides, are a normal feature in birds. Similar thickenings in the few reptiles examined were rich in phospholipids. Extracellular (perifibrous, interstitial) deposits of lipid with a high content of esterified cholesterol were found in more than 90% of human specimens, in 52% of other mammals and in 62.5% of birds included in the survey. In general, lipid insudation of tendons increased with age and was less intense in animals than man. Aetiological factors and the relationship of extracellular lipid deposits in tendons to those in the arterial wall and dura mater are considered.


The smell of cancer.

The smell of cancer. A middle aged man presented reluctantly to his General Practitioner with the complaint of offensive body odour and he was referred for a dermatological opinion. On examination a malignant looking ulcer emitting a foul odour was found in the site of his penis.


Impotence in patients treated with clofibrate. Three of our regularly controlled patients suffering from Type IV hyperlipoproteinemia and treated with clofibrate complained of impotence within one year after commencement of treatment with this drug. Two of the patients had previously suffered from myocardial infarction. Two patients observed improvement of the symptom 3 and 4 weeks after interruption of clofibrate therapy; one patient again complained of impotence when clofibrate therapy was resumed. The third patient continued intake of the drug up to the present day, and still complains of impotence.


The cytological difference between amniotic fluids of male and female fetuses. Counts of cyanophilic squamous cells in 48 specimens of amniotic fluid obtained during the last three months of pregnancy gave the correct diagnosis of the sex of the fetus in every case.  It is confirmed that the fetal genital tract is the source of a considerable exfoliation of cyanophilic squamous cells.  Counts of total cells per mm3 on 38 specimens showed that amniotic fluid from female fetuses at any one week of gestation.  The cytological sex difference should be taken into consideration in the interpretation of differential cell count estimations of fetal maturity.


Role of sex-hormone-binding globulin in hormonal changes and amenorrhoea in thyrotoxic women. Plasma levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), oestrogen, testosterone and sex-hormone-binding globulin (SHBG) were determined in 15 thyrotoxic women in the reproductive age before and after treatment with carbimazole.  The levels of these hormones and SHBG were compared with those of 12 euthyroid healthy female volunteers of comparable age and parity.  Plasma gonadotrophins (LH and FSH), oestrogen, testosterone and SHBG were all markedly elevated in thyrotoxic subjects but became normal when the euthyroid state and normal menstruation were restored after treatment.  From the results a hypothesis is suggested that related the hormonal disturbance primarily to a thyroid hormone-induced rise in SHBG levels.


Use of a specific probe for ovalbumin messenger RNA to quantitate estrogen-induced gene transcripts. DNA complementary to purified ovalbumin messenger RNA (cDNA ov) was synthesized in vitro using RNA-directed DNA olymerase from avian myeloblastosis virus. This cDNAov was then employed in hybridization assays to determine the effect of estrogen on the number of ovalbumin mRNA (MRNAov) molecules per tubular gland cell of the chick oviduct. The changes in mRNAov were measured in immature chicks during primary stimulation, after hormone withdrawal and again following secondary stimulation of the chick oviduct with estrogen. The number  of mRNAov per tubular gland cell was also determined for egg-laving hen. Daily estrogen administration to the immature chick resulted in growth of the oviduct, differentiation of epithelial cells to tubular glands, and a corresponding increase in the concentration of mRNAov in the tubular gland cell from essentially zero before estrogen administration to 48,000 molecules per cell after 18 days of estrogen treatment. Upon withdrawal of estrogen from the chick, the mRNAov concentration decreased to a level of 0-10 molecules/tubular gland cell after 12 days. Readministration of a single dose of estrogen to these chicks resulted in a dramatic and rapid increase in the concentration of mRNAov. Within 30 min, the mRNAov concentration approximately doubled and by 29 hr the tubular gland cell concentration had reached 17,000 molecules. The initial transcription rate for the ovalbumin gene was 12 mRNAov molecules/min. With these data, we have calculated that the half-life of the ovalbumin messinger RNA should be on the order of 40-60 hr and that the steady-state concentration of mRNAov per tubular gland cell was 50,000 molecules. Similarly, each messenger RNA molecule was translated approximately 50,000 times during its lifetime in order to effect the necessary quantity of ovalbumin required for egg production. These data substantiate the hypothisis that estrogen exerts its primary action at the level of transcription to effect the synthesis of nascene mRNA molecules which in turn code for synthesis of hormone-induced proteins.


Stopped-flow kinetic studies of actinomycin binding to DNAs. Stopped-flow kinetic studies of the association of actinomycins with narural and synthetic DNA duplexes are presented. The actinomycins examined were D (C1), D lactam (in which the pentapeptide rings are closed by lactam instead of lactone linkages), X2, XObeta, and actinomine. The DNAs used included claf-thymus DNA, PM2, DNA, and two synthetic d(A-T)-lide copolymers containing 2,6-diaminopurine (DAP) in place of adenine residues, poly[d(DAP-T)]-poly[d(DAP-T)] and poly[d(DAP-A-T]-poly[d(DAP-A-T)]. Apparent equilibrium constants indicate that the DAP-containing polynucleotides bind actinomycin strongly. Comples formation of actinomycins D, D lactam, X2 and XObeta with these DNAs can be deconvoluted into five rate processes. These steps do not necessarily proceed to completion. The rates of two of these steps display a firstorder dependence on DNA concentration. The  large negative entropies of activation of these steps suggest a high degree of restriction to freedom of motion on the respective transition states. The rates of the remaining three steps are independent of DNA concentration. Kinetic parameters of actinimycin binding to DNAs are presented and suggestions are made about some of the molecular evente believed to be responsible for the appearance of the five rate processes. For example, for DNA, poly[d(DAP-A-T)], and poly[d(DAP-T)], the observed order of apparent second-order rate constants, normalized to the concentration of actinomycin binding sites, suggests that binding of the antibiotic occurs most rapidly at binding sites (G-C of d DAT-T) near d(A-T) base pairs, where weakening of the double-helical conformation requires the least energy. Results obtained from studies of actinomycin D binding to heat-denatured poly[d(DAP-A-T)] and of actinomine and actinomycin D lactam binding to DNA suggest that the slow rate processes are related to an actinomycyl-pentapeptide-induced unwinding of the sugar-phosphate backbone of DNA accompanying insertion of the cyclic peptides into DNA.


Human myeloma IgG half-molecules. Structural and antigenic analyses,. The structure and antigenic characteristics of a human k, IgG myeloma protein that formed half-molecules were analyzed. Most of the myeloma protein found in the patient's serum and urine consisted to two chain 4.3S half-molecules. A small amount of four chain 7S myeloma protein was, however, found in the serum and was apparently formed by the same clone of tumor cells. Polyacrylamide gel electrophoresis in 8 M urea and 1% sodium dodecyl sulfate and analytical ultracentrifugation in 6 M guanidine of the fully reduced and alkylated half-molecule indicated that this myeloma protein had a heavy chain of a smaller molecular weight (approximately 45,000) than that of normal gamma chains, Except for this apparent deletion, the heavy chain resembled gamma1 chains. The amino acid composition of the peptides containing the half-cysteine residues forming the interchain disulfide bonds, the glycopeptide of the Fc fragment and the COOH-terminal structure were similar if not identical with the analogous structures of gamma1 chains. No Fc fragment could be prepared because the Fc portion of the heavy chain of the myeloma protein was extremely susceptible to degradation with papain. After mild reduction and alkylation, the 7S myeloma protein dissociated into half-molecules, indicating a lack of noncovalent interactions in the Fc fragment that are present in all classes of human immunoglogulins and are responsible for the formation ofFc dimers. The half-molecule was antigenically deficient in the Fc fragment. It failed to precipitate with anti-Fc fragment antisera in double gel diffusion tests and inhibited a Fc-anti-Fc fragment binding reaction weakly and incompletely. The half-molecule and the 7S protein had the same genetic markers on the first and second homology region of the gamma chain. The half-molecule lacked, however, the corresponding markers on the third homology region, These findings suggest that this myeloma protein had a deletion in the gamma chain which was probably located in third homology region and was likely the structural abnormality responsible for the lack of noncovalent interaction in the Fc fragment and absence of most of the antigenic determinants characteristic of gamma chains.


Conformational and spectral analysis of the polypeptide antibiotic N-methylleucine gramicidin S dihydrochloride by nuclear magnetic resonance. The 220-MHz proton magnetic resonance spectrum of the cyclic decapeptide antibiotic, mono-N-methylleucine gramicidin S, is reported and all the resonances have been assigned to specific protons of the constituent amino acids. Three methods--temperature dependence and solvent mixture (methanol-trifluoroethanol and dimethyl sulfoxide-trifluoroethanol) dependence of peptide NH proton chemical shifts and proton deuteron exchange--habe been utilized to delineate peptide NH protons. The results of the above methods, coupled with the observed vicinal alpha-CH-NH coupling constants and chemical shifts, indicate that in trifluoroethanol the peptide NH PROTONS OF D-Phe4, D-Phe9, L-Orn2, and L-Val6 are exposed to the sovent, and those of L-Val1, L-Orn7, and L-Leu8 are solvent shielded and intramolecularly hydrogen bonded. In trifluoroethanol, dimethyl sulfoxide, and methanol, the decapeptide has no C2 symmetry, and there are only minor conformational differences in the different solvents. In the proposed conformation in trifluoroethanol, one-half of the decapeptide retained the hydrogen bonding pattern of gramicidin S, i.e. cyclo-(L-Val1 NH--O-C L-Leu8) (a beta turn) and cyclo-(L-Leu8 NH--O-C L-Val1). The second half of the molecule exhibits a different type of stable beta turn involving the ten-atom hydrogen-bonded ring, cyclo-(L-Orn7-NH--O-C D-PHE4).


Reactions of fluorescent probes with normal and chemically modified myelin basic protein and proteolipid. Comparisons with myelin. Basic (encephalitogenic) protein and water-soluble proteolipid apoprotein isolated from bovine brain myelin bind 8-anilino-1-naphthalenesulfonate and 2-p-toluidinylnaphthalene-6-sulfonate with resulting enhancement of dye fluorescence and a blue-shift of the emission spectrum. The dyes had a higher affinity and quantum yield, when bound to the proteolipid (Kans=2.3x10--6,=0.67) than to the basic protein (Kans=3.3x10--5,=0.40). From the efficiency of radiationless energy transfer from trytophan to bound ANS the intramolecular distances were calculated to be 17 and 27 A for the proteolipid and basic protein, respectively. Unlike myelin, incubation with proteolytic enzymes (e.g., Pronase and trypsin) abolished fluorescence enhancement of ANS or TNS by the extracted proteins. In contrast to myelin, the fluorescence of solutions of fluorescent probes plus proteolipid was reduced by Ca-2+,not affected by La-3+, local anesthetics, or polymyxin B, and only slightly increased by low pH or blockade of free carboxyl groups. The reactions of the basic protein were similar under these conditions except for a two- to threefold increase in dye binding in the presence of La-3+, or after blockade of carboxyl groups. N-Bromosuccinimide oxidation of tryptophan groups nearly abolished native protein fluorescence, but did not affect dye binding. However, alkylation of tryptophan groups of both proteins by 2-hydroxy (or methoxy)-5-nitrobenzyl bromide reduced the of bound ANS (excited at 380 nm) to 0.15 normal. The same effect was observed with human serum albumin. The fluorescence emission of ANS bound to myelin was not affected by alkylation of membrane tryptophan groups with the Koshland reagents, except for abolition of energy transfer from tryptophan to bound dye molecules. This suggests that dye binding to protein is negligible in the intact membrane. Proteolipid incorporated into lipid vesicles containing phosphatidylserine did not bind ANS or TNS unless Ca-2+, La-3+, polymyxin B, or local anesthetics were added to reduce the net negative surface potential of the lipid membranes. However, binding to protein in the lipid-protein vesicles remained less than for soluble protein. Basic protein or bovine serum albumin dye binding sites remained accessible after equilibration of these proteins with the same lipid vesicles. It is proposed that in the intact myelin membrane the proteolipid is probably strongly associated with specific anionic membrane lipids (i.e., phosphatidylserine), and most likely deeply embedded within the lipid hydrocarbon matrix of the myelin membrane. Also, in the intact myelin membrane the fluorescent probes are associated primarily, if not solely with the membrane lipids as indicated by the binding data. This is particularly the case for TNS where the total number of myelin binding sites is three to four times the potential protein binding sites.


Plasma proteins and immunoglobulins in penguins (Pygoscelis adeliae). Total and fractionated proteins were evaluated in the serum of normal penguins. Four proteins with antibody activity, determined by passive haemagglutination, were isolated by Sephadex G-200 filtration and DEAE-cellulose chromatography, from the serum of penguins (Pygoscelis adeliae) inoculated with dinitrophenylated human gamma-globulin. Immunoelectrophoresis and immunodiffusion showed that all these proteins exhibited both their own and shared antigenic determinants. On account of their electrophoretic mobility, elution conditions and characteristics of the precipitation arcs, they have been tentatively denominated IgG2 (gamma2), IgG1 (gamma1), IgM (gammam) and IgX (probably IgA).


Analysis of the conformation of polypeptides : the combined use of energy computations and nuclear magnetic resonance studies. This review surveys current approaches to the problem of determining the solution conformation of polypeptides. The basic principles of energy computations are described. The utility, problems, and limitations of various theoretical methods are summarized: conformational energy mapping, energy minimization, scanning of selected local conformations, statistical predictive schemes. The need for combining the calculations with experimental studies is pointed out. The information content of various physico-chemical methods is compared for this purpose. The analysis of nmr coupling constants is discussed in more detail. In combination with energy computations, it can furnish specific information on local aspects of the conformation. Examples of such combined studies on small peptides are summarized.


Binding of 14-C-salicylic acid and 14-C-pentobarbital to plasma proteins of several species during the perinatal period. The fraction binding of 14-C-salicylic acid and 14-C-pentobarbital was studied as a function of age in plasma of the pig, dog, goat and human. The pig exhibited an unusual degree of hypoalbuminemia and low fractional binding of salicylic acid at birth. Albumin levels and the percent binding of the salicylic acid were not significantly lower in newborn plasma than in adult plasma of the other species studied. The pig was also exceptional in that the high fractional binding of pentobarbital observed at low ligand concentrations in plasma of the newborn was not observed in the other species studied. Depletion of albumin in newborn pig plasma did not appreciably affect the binding of pentobarbital. Thus it is suggested that the plasma of the fetal and newborn pig may contain a nonalbuminoid protein, such as fetal globulin, which is capable of binding pentobarbital but does not have a high avidity for salicylic acid. It apparently does not exist at birth in the plasmas of the other species studied, as depletion of albumin markedly reduced the fractional binding of pentobarbital.


IgM deficiency.

IgM deficiency. Primary isolated IgM deficiency accounted for 0.1% of hospital admissions; secondary IgM deficiency for 2.0%. Although 19% were asymptomatic the rest of 89 subjects (4M:1F) suffered infection (60%), septicemia (36%), atopy (22%), splenomegaly (11%), neoplasia(7%) and autoimmune disorders (3%) with a mortality of 10%. Serious early treatment is needed to avert death from unopposed spread of organisms throughout the blood. Qualitative IgM deficiency (absence of isohemagglutinins) and delayed maturation of IgM can result in similar symptomatology.


Autoimmunity in selective IgA deficiency. Several antibodies have been reported in selective IgA deficiency and we have detected antibodies to human IgM and native human collagen in 35 and 34%, respectively, of 60 such subjects (compared to 1.6 and 2.0%, respectively, in normal controls). The anti-IgM isoantibodies are clearly distinguishable from antibodies to ruminant proteins and there was no statistically significant association between different antibodies and clinical features in individual patients. The etiology of the antibodies remains unknown.


Lymphocytotoxins and immunologic unresponsiveness. A 14-year-old boy with a lifelong history of recurrnet infections and debilitating bronchiectasis was found to lack any evidence of humoral or cellular immunity. His serum contained a high titer of IgM antibody to the heavy chain of IgG and this antibody was also cytotoxic for peripheral lymphocytes. Complement-dependent lymphocytotoxicity could be blocked by IgG or Fc fragment. In this patient, immunosuppression may have been due to an autoantibody against both autologous IgG and lymphocyte plasma membrane.


The monoclonal nature of lymphatic leukemia. Monoclonal membrane-bound Ig were found by immunofluorescence on the lymphocytes in the vast majority of cases of chronic lymphocytic leukemia. The distribution of H and L chains among these patients reflected the distribution of surface Ig on normal lymphocytes and IgM was the predominant class. The importance of the study of surface Ig synthesized in vitro is outlined. The simple staining of freshly drawn cells may lead to erroneous conclusions, since an apparently polyclonal staining can result from the anti-IgG antibody activity of a monoclonal surface IgM, from the attachment of immune complexes at the lymphocyte surface or from the binding of serum antibodies to cell membrane determinants. A biclonal proliferation, characterized by distinct surface-Ig markers, was demonstrated in several cases of chronic lymphatic leukemia. Monoclonal surface Ig were also detected on the lymphoblasts in 2 cases of acute transformation of chronic lymphocytic leukemia, in several patients with acute lymphosarcoma cell leukemia and in 1 case of acute lymphatic leukemia. In most cases of acute lymphatic leukemia, the leukemic cells were devoid of detectable B- or T-cell membrane markers. In 2 cases of acute lymphatic leukemia, 1 case of chronic lymphatic leukemia and in all patients with the Sezary syndrome, the leukemic cells appeared to be thymus-derived.


Conventional immunoglobulin therapy.

Conventional immunoglobulin therapy. Concentrated gamma globulin as a substitute for deficient B-cell function is usually given by the intramuscular route. The intravenous administration would be preferable, but presents major difficulties, mainly with shock-life reactions. Most of them are due to rapid activation of the complement system by aggregated IgG. The attempts to avoid such reactions include exposure of gamma globulin concentrates to pepsin, plasmin, low pH, beta-propiolactone, polymers, salts and millipore filtration. The results improved slowly, but they are still not entirely satisfactory.


B lymphocytes lacking surface IG in patients with immune deficiency: initiation of IG synthesis in culture in cells of a patient with thymoma. A 64-year-old woman with a syndrome of thymoma, severe hypogammaglobulinemia, seemingly normal cell-mediated immunity and aplastic anemia, was found to have virtually no immunoglobulin- (Ig) bearing peripheral blood lymphocytes (PBL). However, 7.8 +/- 3.4% of the PBL were positive for another B-cell marker, the receptor for aggregated IgG, while the remaining cells bound sheep erythrocytes. Those cells which were aggregate-reactive appeared to be immature or incomplete B cells. Cultures of peripheral blood leukocytes from the patient in various serum-containing media were studied by 3 independent technics for the development of lymphocyte surface Ig and for Ig in the culture supernatants. In vitro the patient's cells were able to develop surface Ig in media supplemented with fetal calf serum (FCS) or normal serum; in media supplemented with autologous serum, the cells developed no surface Ig. During the cultures in FCS, human Ig determinants became detectable in the medium, and both medium and cell-surface Ig underwent a shift from mu determinants early in the culture period to gamma and alpha determinants later. The development of Ig on the cells was not inhibited by the presence of autologous serum if FCS was included in the medium. These data support the concept that a factor, missing from this patient's serum, is required at an early stage in the maturation of the B cell. A patient with X-linked agammaglobulinemia had a population of circulating lymphocytes with surface characteristics similar to the B cells of the thymoma case. In contrast, no Ig synthesis by this patient's cultured cells could be demonstrated, indicating a different level of block in the 2 cases despite their similarity at the level of the cell surface.


Lymphocyte transformation in vitro in patients with immunodeficiency diseases: use in diagnosis, histocompatibility testing and monitoring treatment. We have used lymphocyte stimulation in vitro to characterize the degree of cell-mediated immunodeficiency in different patients. The effect of treatment of patients with immunodeficiencies is illustrated by lymphocyte transformation in vitro before, during and after therapy (eg bone marrow transplantation of severe combined immunodeficiency and transfer factor treatment in Wiskott-Aldrich syndrome). The mixed lymphocyte culture test has been used for selection of bone marrow transplant donor for a patient with CID when an HL-A identical sib was not available. The results of the transplantation in this patient with graft from a donor who differed from the patient in respect of both HL-A haplotypes (but MLC identical with the patient) is reported. The MLC data on another family with 2 children with Nezelof disease are reported. The data indicate that one of the patients could be a chimera after intrauterine grafting of maternal immunocompetent cells.