Aortic insufficiency requiring valve replacement in Whipple's disease. Reviews of postmortem reports on patients with Whipple's disease (intestinal lipodystrophy) describe gross valvular deformity in more than 50% with characteristic histological findings of macrophages containing periodic acid-Schiff-positive, diastase-resistant granules. Frequently, congestive heart failure characterizes the terminal stages. In a 58-year-old man with well-documented Whipple's disease for 5 years, gastrointestinal, joint, and pericardial involvement apparently resolved with medical therapy. However, 10 years later, severe aortic insufficiency necessitated prosthetic valve replacement, at which time gross and histological examination of the excised valve demonstrated characteristic changes of Whipple's disease. Clinical recognition of the importance of cardiac valvular abnormalities and of possible late cardiac decompensation mandates close observation of patients with Whipple's disease. Corrective operation should improve the patient's chances of survival.


Kinetic haemagglutination inhibition technique as a means of detecting antigenic variations among strains of Nigerian flaviviruses. Seven isolates of West Nile virus and an additional isolate tentatively identified at the outset as West Nile virus, were compared by Casals' modified haemagglutination inhibition technique. Two of the isolates were nearly identical; a second set of five isolates cross-reacted among themselves within a wide range and differed from the first two; the last isolate was significantly different from the others.


Antigenic differences between strains of Newcastle disease virus. Comparison of strains of Newcastle disease virus by kinetic neutralization tests and immunodiffusion tests (after virion disruption by ether and Tween 80) enabled antigenic differences to be demonstrated between four out of five strains tested. No correlation was demonstrated between the antigenic structure and virulence of these strains.


[Method of intravital injection of the internal blood vessels of organs for experimental-morphological research]. Ten minutes before the injection the rat is given heparin (500 units per 1 kg weight) intradermally. To the narcotized animal, the needle with the 1 ml syringe is inserted and 3--5 ml of blood are aspirated (about 2% of the body mass). Then with the same needle another syringe of 10 ml volume containing filtrated and heated up to 40 degrees C undiluted Indian ink is injected. The amount of the Indian ink injected is 8--10 ml per 100 g of the body mass. We can consider the injection oa success if the mucose of the tongue, the skin of the concha auriculae, the sclera of the eye are promptly stained during the injection and the tail vessels are well filled.


[Differential staining of pancreatic islet A- and B-cells using Rouge's fixative and a modified Gomori method]. Roug's fixative fluid is suggested to fix the pancreas with subsequent staining of the islet A- and B-cells. Rouget's fluid differs from Bouin's fluid in that picric acid is substituted for distilled water, and thus, it is unnecesary to extract the acid from the preparation. The fixation ensures good quality in demonstration specific granulation and preserves it in B-cells. Gormori's modification of the method to stain the slices with aldehyd-fuchsin and subsequent staining with alum carmine and orange G reveals both B- and A-cells. The manipulations presented perfect the quality of histochemical reactions.


[Polychromatic method of simultaneously demonstrating all the structural components of a peripheral nerve]. To study neuro-vessel relations in the nerve conductive system, a combined method revealing myelinated and non-myelinated nerve fibers and blood vessels was suggested. The method includes the following stages of material treatment: injection of the nerve blood vessels with chloroform emulsion of Paris blue (5--10 g of the dye in 100 g of the solvant); the straightened nerves about 5 mm long are fixed in 12% neutral formalin; etching in the solution of chromium solts; embedding in paraffin and preparation of thin sections (3--5 mcm); staining after Masson's method applied for collagen fiber staining) in the author's modification and mounting in balsam. In the preparations, the axons of the nerve fibers are seen blue--purple, myelin sheaths--light red, connective tissue elements--violet and blood vessels are seen as dark blue rings.


[Method of staining cation proteins of the polymorphonuclear leukocyte lysosomes in the histological paraffin sections]. A method has been proposed for detection of lysosomal cation proteins of polymorphonuclear leukocytes in histological paraffin sections. The method is based on staining of histological sections of Brank fluid-fixed material with an alcohol solution of green fast at pH 8.1. A 0.25% aqueous solution of azure A was used for poststaining of cell nuclei and viable bacteria.


[Morphological characteristics of ground glass hepatocytes in chronic hepatitis and liver cirrhosis]. Histochemical and electron microscopic examinations of liver punctates from 57 patients with chronic hepatitis and cirrhosis of the liver were carried out. "Ground Glass" hepatocytes were found to occur with similar frequency both in patients with HB-antigen in the blood serum and in patients with alcoholic involvement of the liver and no antigen in the blood serum. The presence of HB-antigen in the cytoplasm of such hepatocytes can be detected by staining of the sections with orsein, aldehydethionine and aldehydefucsin. The latter stain is more specific. Electron microscopically, the "Ground Glass" hepatocytes are characterized by marked hyperplasia of agranular endoplasmic reticulum in the lumen of which peculiar filaments are found in the presence of HB-antigen.


[HB-antigenemia and liver pathology].

[HB-antigenemia and liver pathology]. The review deals with liver pathology in asymptomatic carriers of hepatitis B antigen. Information on the pattern and frequency of liver affection in carriers of HB antigen, on morphological methods for the detection of HB antigen in the liver tissue, on the frequency of the antigen detection in the liver tissue in asymptomatic antigen carriers and in liver diseases with HB antigenemia is presented. The data concerning the clinical and prognostic importance of hepatitis B antigen detection are given.


Proteolytic and peroxidatic reactions of commercial horseradish peroxidase with myelin basic protein. Degradation of myelin basic protein during incubations with high concentrations of horseradish peroxidase has been demonstrated [Johnson & Cammer (1977) J. Histochem. Cytochem.25, 329-336]. Possible mechanisms for the interaction of the basic protein with peroxidase were investigated in the present study. Because the peroxidase samples previously observed to degrade basic protein were mixtures of isoenzymes, commercial preparations of the separated isoenzymes were tested, and all three degraded basic protein, but to various extents. Three other basic proteins, P(2) protein from peripheral nerve myelin, lysozyme and cytochrome c, were not degraded by horseradish peroxidase under the same conditions. Inhibitor studies suggested a minor peroxidatic component in the reaction. Therefore the peroxidatic reaction with basic protein was studied by using low concentrations of peroxidase along with H(2)O(2). Horseradish peroxidase plus H(2)O(2) caused the destruction of basic protein, a reaction inhibited by cyanide, azide, ferrocyanide, tyrosine, di-iodotyrosine and catalase. Lactoperoxidase plus H(2)O(2) and myoglobin plus H(2)O(2) were also effective in destroying the myelin basic protein. Low concentrations of horseradish peroxidase plus H(2)O(2) were not active against other basic proteins, but did destroy casein and fibrinogen. Although high concentrations of peroxidase alone degraded basic protein to low-molecular-weight products, suggesting the operation of a proteolytic enzyme contaminant in the absence of H(2)O(2), incubations with catalytic concentrations of peroxidase in the presence of H(2)O(2) converted basic protein into products with high molecular weights. Our data suggest a mechanism for the latter, peroxidatic, reaction where polymers would form by linking the tyrosine side chains in basic-protein molecules. These data show that the myelin basic protein is unusually susceptible to peroxidatic reactions.


Measurement of H-2 antigen and immunogenicity of methylcholanthrene-induced murine sarcomas. For each of a set of 11 methylcholanthrene-induced sarcomas of B10 mice, we measured the amount of H-2 antigen by absorption of a specific antiserum, and the strength of the tumour-specific transplantation antigen by a transplantation assay, to see whether they are correlated. No obvious correlation was seen. We showed that cell suspensions of tumours taken directly from the animal are contaminated by host cells which make a substantial contribution in H-2 assays. Since this contamination was lost after several passages in vitro, the amount of H-2 on tumour cells was assayed only after such passages.


Circulating levels of alpha-fetoprotein and pregnancy specific beta1 glycoprotein in pregnancies without an embryo. Ten patients with depressed serum alpha-fetoprotein (AFP) levels at 14 to 15 weeks gestation were found to have no embryo on ultrasonography: two patients had hydatiform moles and the remainder had anembryonic pregnancies (blighted ova). The pregnancy specific beta1 glycoprotein (SP1) levels in serum were measured in all these ten patients. Seven of the eight patients with anembryonic pregnancies and one of the two patients with molar pregnancies had low serum SP1 levels. These observations suggest that the measurement of AFP and SP1 may provide biochemical evidence of pregnancy without an embryo.


Existence of common antigenic sites in tropomyosins. 1. Tropomyosins were extracted from vertebrate and invertebrate muscles, and their immunolo;ical characteristics were compared using antisera against tropomyosins from chicken skeletal and cardiac muscles. 2. Antigenic sites common to those of chicken skeletal muscle tropomyosin were found in all the tropomyosins tested, although the reactions of these common antigenic sites in an immunodiffusion test were weak in tropomyosins from phylogenetically distant animals. 3. An immunological difference was found between alpha-tropomyosins from chicken cardiac muscle and rabbit cardiac muscle. Thus they had specific antigenic sites in addition to the common ones. 4. A component was found in a 1 M KCL extract of Tetrahymena pyriformis which reacted with antiserum against chicken skeletal muscle tropomyosin.


Fluorescence studies of the interactions of serum albumin and rat alpha1-fetoprotein with aflatoxin B1. Specificity and binding parameters. The interaction of bovine serum albumin and rat alpha1-fetoprotein with aflatoxin B1 has been followed by the fluorescence quenching of the protein in presence of the ligand. The binding parameters (n, number of sites and Kd, dissociation constant) have been determined for the bovine serum albumin-alflatoxin B1 system: n = 3.5 and Kd = 3.1 +/- 0.5 . 10(-5) M and for the alpha-fetoprotein-aflatoxin system: n = 4 and Kd = 3.7 +/-0.5 . 10(-5) M. The competition of anilino-naphthalene-sulfonate and aflatoxin B1 for the same hydrophobic sites on bovine serum albumin has been demonstrated. The fluorescence quenching of various proteins (lysozymes, egg-albumin, gamma-globulin) by aflatoxin B1 have shown that there is not a strict specificity of aflatoxin towards proteins.


Oestrone binding properties of nine molecular variants of rat alpha-foetoprotein. Rat alpha-foetoprotein was separated into nine molecular variants by electrophoresis and affinity chromatography on Ricinus communis agglutinin and concanavalin-A. The nine variants are able to bind oestrone with the same capacity of one binding site per alpha-foetoprotein molecule. The association constants seem to vary with the sialic acid composition of the iso-alpha-foetoprotein.


Inhibition of the RNA dependent DNA polymerase and the malignant transforming ability of Rous sarcoma virus by thiosemicarbazone-transition metal complexes. Several thiosemicarbazone-metal complexes inhibit the RNA dependent DNA polymerase and the transforming ability of Rous sarcoma virus. Some complexes are equally as active as the free ligand whereas the activity of others is greatly enhanced. The 2-formyl pyridine thiosemicarbazone copper (II) complex is the most potent compound of this class that we tested. Some copper complexes of salicylaldehyde derivatives are very active also, particularly N-n-butyl, N-n-hexyl and N-benzylsalicylaldimine; no nickel complex of any salicylaldehyde compound is active. In addition, other metal ligands, such as dithizone, diacetyl bis (mercaptoethylimine), N-butyl thiocarbamate, 0,0' dimethyl dithiophosphate, potassium dithiooxalate, and cis-PtII(NH3)2Cl2 were tested with varying results.


Coordination behaviour of antithyroid drugs against the Fe(I)(NO)2 group in solution: ESR and FT-NMR study. The binding sites of some types of antithyroid drugs in the presence of the Fe(I)(NO)2 paramagnetic probe were investigated. Coordination behaviour in solution of different structured ligands was determined by means of ESR parameters and 13C FT-NMR spectra. Selective isotopic substitution with 15NO combined with computer simultation was used to elucidate overlapping ESR patterns. A correlation between chemical structure and antithyroid activity of the pharmacological bases is suggested.


Fluorescence polarization of stretched polytene chromosomes stained with acridine orange. The molecules of the fluorescent dye acridine orange (AO) bind to DNA in such a way that the absorption and emission dipoles lie on a plane perpendicular to the DNA axis. For this reason, definite fluorescence polarization should correspond to each mode of spatial DNA packing. A chromosome, considered as an axially symmetrical ensemble of DNA, was characterized by two experimental parameters, P parallel and P perpendicular, i.e., by polarizations of fluorescence excited by light polarized parallel and perpendicular to the symmetry axis. In view of the sequential order in the packing levels of DNA fiber in a chromosome, it was suggested that, under mechanical stretching, the highest level is disrupted first, then the others, in the order of their sequence. Isolated chromosomes of Chironomus thummi were stained with AO and stretched with needles of a micromanipulator. From the changes of P parallel and P perpendicular measured during stretching it was concluded the polytene chromosome bands have three, at least, DNA packing levels, tentatively described as 100 A fiber, 250 A coil and chromomere.


[Concentration of specific antibodies in commercially avaialable immunoglobulins (author's transl)]. Not significant differences in the composition or concentration of specific antibodies against microbial antigens could be measured in five commercially available human immunoglobulin preparations for intravenous use. Prediction of prophylactic or therapeutic efficiency of such preparations according to their antibody content seems to be only partially possible. Human immunoglobulins for intravenous use should be free from irregular antibodies against erythrocyte antigens of the Rh-systems, as found in one of the specimens tested.


Mesencephalic and diencephalic afferents to the superior colliculus and periaqueductal gray substance demonstrated by retrograde axonal transport of horseradish peroxidase in the cat. The mesencephalic and diencephalic afferent connections to the superior colliculus and the central gray substance in the cat were examined by means of the retrograde transport of horseradish peroxidase (HRP). After deep collicular injections numerous labeled cells were consistently found in the parabigeminal nucleus, the mesencephalic reticular formation, substantia nigra pars reticulata, the nucleus of posterior commissure, the pretectal area, zona incerta, and the ventral nucleus of the lateral geniculate body. A smaller number of cells was found in the inferior colluculus, the nucleus of the lateral lemniscus, the central gray substance, nucleus reticularis thalami, the anterior hypothalamic area, and, in some cases, in the contralateral superior colliculus, Forel's field, and the ventromedial hypothalamic nucleus. Only the parabigeminal nucleus and the pretectal area showed labeled cells following injections in the superficial layers of the superior colliculus. In the cats submitted to injections in the central gray substance, labeled cells were consistently found in the contralateral superior colliculus, the mesencephalic reticular formation, substantia nigra parts reticulata, zona incerta and various hypothalamic areas, especially the ventromedial nucleus. In some cases, HRP-positive cells were seen in the nucleus of posterior commissure, the pretectal area, Forel's field, and nucleus reticularis thalami. A large injection in the mediodorsal part of the caudal mesencephalic reticular formation, which included the superior colliculus and the central gray substance, resulted in numerous labeled cells in nucleus reticularis thalami. The findings are discussed with respect to the suggested functional division of the superior colliculus into deep and superficial layers. Furthermore, the possible implications of labeled cells in zona incerta and the reticular thalamic nucleus are briefly discussed.


The response of cell walls of Bacillus subtilis to metals and to electron-microscopic stains. Purified cell walls of Bacillus subtilis were subjected to solutions of 40 independent metals and the metal uptake, the electron-scattering power of thin sections, and the type of staining response evaluated. This was repeated for six typical electron-microscopic stains (uranyl acetate, uranyl magnesium acetate, osmium tetroxide, Os-meth, osmium-dimethylethylenediamine, and ruthenium red) and one new staining reagent (a potassium platinum chloride - dimethylsulfoxide complex) whose specificity is for amine functions. The reaction of select metals can be specific in terms of both uptake and staining response. Of the metals studied most transition elements had a high affinity for the wall fabric and some (i.e., Sc III, most lanthanides, UIV, ZrIV,HfIV, Fe III, Pd II, Ru III, and In III) may be suitable as contrasting agents for electron microscopy. Furthermore, when the thickness of metal-reacted walls was compared to freeze-each and ultracryotomy data, statistical-dimensional differences were commonly seen, which indicates that wall ultrastructure can be profoundly affected by the type of metal and (or) staining reagent.


Treatment of advanced Hodgkin's disease with B-CAVE following MOPP failure. Between March 1973, and December 1976, 22 patients who developed disease progression during or after MOPP therapy were treated with a new combination, B-CAVe (Bleomycin 5 mg/m2 iv days 1, 28, 35; CCNU 100 mg/m2 po day 1; adriamycin 60 mg/m2 iv day 1; and vinblastine 5 mg/m2 iv day 1). Objective responses were achieved in 17 of 22 patients (77%) and 11 of 22 responses were complete (50%). The actuarial survival for all patients is 16.4 months. For complete responders the median is 24 months with 2 complete responders dead without evidence of Hodgkin's Disease. Median relapse free survival for complete responders has not been reached at 35+ months while that for partial responders is 14 months. Significant adriamycin cardiotoxicity was encountered in two patients. There were no life threatening bacterial infections during B-CAVe. Two patients died of Pneumocystis carinii several months after cessation of therapy. B-CAVe is effective in the therapy of advanced Hodgkin's disease after MOPP failure, and this regimen is comparable to other previously reported MOPP salvage combinations.


Melanoma skin test antigens of improved sensitivity prepared from vesicular stomatitis virus-infected tumor cells. Crude membrane (CM) extracts from three different cultured human melanoma lines that were "virus-augmented" (infected with vesicular stomatitis virus (VSV) and subsequently inactivated by ultraviolet light) produced positive skin tests in 17 of 20 (85%), 11 of 20 (55%), and 13 of 18 (72%) tests, respectively, performed in 20 melanoma patients. Identical CM extracts from the same melanoma lines that had not been infected with VSV gave positive skin tests in 2 of 20 (10%), 4 of 20 (20%), and 2 of 18 (11%) tests, respectively, performed in the 20 melanoma patients, and no positive tests in the control patients. The 3 virus-augmented extracts were positive in only 2 of 18 (11%), 0 of 18 (0%), and 1 of 17 (6%) control subjects, respectively. The controls consisted of six normal volunteers and 12 patients with cancers other than melanoma. The "virus-augmented" CM extracts thus exhibited markedly greater sensitivity without significant loss of specificity as compared to nonvirus augmented extracts when used as tumor-specific melaonma skin test antigens.


Basal cell proliferation in benign prostatic hyperplasia. Two cell types are found within glandular acini of the human prostate. These basal and secretory cells can be distinguished in the light microscope by their different affinity for toluidine blue. Toluidine blue exhibits a strong affinity for basal cells while secretory cells are poorly stained. In benign prostatic hyperplasia, toluidine blue stained basal cells commonly form a single layer which rests on the basement membrane of glands. Also present are areas where cell layers located nearer gland lumina, thought to be composed of secretory cells, are occupied by cells with a basal like morphology and which exhibit a strong affinity for toluidine blue. The ultrastructure of these cells resembles that of basal cells. Autoradiographic analysis of 3H-thymidine incorporations by hyperplastic prostatic expants in organ culture shows that within glands only basal cells are active in DNA synthesis. The data suggest that basal cells are capable of proliferating and may be a source of secretory cells.


A cytologic study on small cell carcinoma of the esophagus. The cytologic picture of small cell carcinoma primarily arising from the esophagus was studied with 7 cases which were confirmed by histologic examination. Cytomorphologic characteristics of small cell carcinoma of the esophagus are as follows: the arrangement of groups of tumor cells is irregular and overlapping with indistinct cell boundaries. The cytoplasm is small, or sometimes absent. The nuclei are round, oval or occasionally spindle shaped. Nuclear borders are thin. The chromatin of finely granular pattern has increased and is evenly distributed.


Palliation of hepatic metastasis.

Palliation of hepatic metastasis. Between 1971 and 1975, 55 patients underwent palliative radiation therapy for symptomatic hepatic metastasis. Most patients received 2400 rad in 300 rad fractions to the entire liver. There were 31 patients who received concomitant chemotherapy, and 14 who were prior chemotherapy failures. Ninety percent of the patients with symptomatic pain and liver enlargement and significant palliation of their symptoms. The median survival of the entire group was 4.5 months, while those patients experiencing an excellent response (21) had a median survival of 9 months. The median survival of patients having an excellent response to radiation is comparable to that of patients having regional arterial chemotherapy while incuring fewer complications. The overall complication rate of those patients completing therapy (50) was 12%.


A combined assay for the rapid preliminary detection of structural retroviruses. This report describes the use of equilibrium gradients, RNA dependent DNA polymerase assays and electron microscopy (EM) in a combined assay for the rapid preliminary detection of intact retroviruses in crude preparations. Positive combined assays of platelets from preleukemic patients corresponded with karyotypic abnormalities found in these patients. Reconstruction experiments with Rauscher Leukemia Virus added to buffer or disrupted mouse spleen demonstrated the ease of detecting 10(9) or greater particles/g crude tissue, and the effects of buffer or added protein.


Maytansine action on fast axoplasmic transport and the ultrastructure of vagal axons. Maytansine, an ansa macrolide now in clinical trials as an antineoplastic drug, is a potent inhibitor of microtubule polymerization. Since microtubules are involved in axoplasmic transport, the effect of maytansine on transport was examined. Fast axoplasmic transport of proteins and the axonal ultrastructure was studied in the vagus nerve of cats exposed in vitro to maytansine. Tritiated leucine was microinjected into the nodose ganglion; after 2 hr for incorporation into proteins, nerves were dissected out for transport and ultrastructural studies and incubated for 2.5 hr in Krebs-Ringer solution with 100, 20, 10, 5, or 1 micron maytansine. A reduction in the number of microtubules and a partial blockage of fast axoplasmic transport was observed at 20 and 100 micron maytansine; at 10 micron no detectable changes were observed. These findings show that maytansine in vitro induces alterations of the neurofibrillar elements concomitant with a partial blockage of fast axoplasmic transport.


Epidermla cell kinetics in hairless mice after bleomycin. I. Perturbations after different single doses. Three different doses of bleomycin (BLM) (2,1, and 0.01 mg/animal) were injected ip into groups of hairless mice. Cell kinetic alterations in the basal layer of the epidermis were studied up to 10 days after BLM administration. The two highest doses of BLM affected the epidermal cell kinetics in a similar way but with different time courses. Only a moderate depression of the labeling index was observed during the first 24 hours. The results indicate that cells affected by BLM in the middle of the G1 phase are arrested in their subsequent S phase, and that thereafter an increase in the rate of DNA synthesis may occur in the arrested cells without increasing the number of cells in S phase. The percentage of cell accumulated in S and G2 phases as determined by flow cytofluorometry never exceeded 130% of control values with any of the doses. Phase durations were determined from percentage of labeled mitoses curves after the highest dose of BLM and were not significantly different from normal values, indicating that the accumulated cels are probably out of cycle and unable to proliferate again. Calculations based on cell counts and mitotic rate showed that the cells had a prolonged lifespan. No block and subsequent release of cells specifically caused by BLM were observed. BLM has a complex effect on the epidermis, and therefore does not seem to be a promising agent for cell synchronization as a basis for combination chemotherapy in vivo.


Cyclocytidine chemotherapy for malignant melanoma. Twenty-nine patients with metastatic malignant melanoma were treated with cyclocytidine, 240 mg/m2/day sc for 10 days. All partients had received extensive prior chemotherapy. Only one patient achieved a partial remission; the overall response rate (complete plus partial) was 4%. Unusual toxic effects associated with cyclocytidine chemotherapy included the delayed onset of thrombocytopenia, orthostatic hypotension, and jaw pain.


Use of prostaglandin E1 for emergency palliation of symptomatic coarctation of the Aorta. Prostaglandin E1 (PGE1) was infused into the pulmonary artery in a 1-day-old infant with symptomatic coarctation. Umbilical artery pressure rose from 50/40 mm Hg before infusion to 83/48 mm Hg after infusion, indicating significant improvement in lower-body perfusion. Successful coarctation repair was performed during continued PGE1 infusion. To our knowledge, this is the first reported patient in whom PGE1 has been used for successful emergency palliation of symptomatic coarctation.


Liver growth, biosynthesis of cytidine nucleotides and level of cytochrome P-450 in rat liver after administration of alpha-hexachlorocyclohexane. The biosynthesis of cytidine nucleotides and the level of microsomal cytochrome P-450 in intact and regenerating rat liver after repeated administration of alpha-hexachlorocyclohexane (alpha-HCH) were compared. In alpha-HCH treated animals the utilization of [2-14C] orotic acid for the synthesis of cytidine nucleotides is suppressed. In 24-h regenerating liver the incorporation of labelled orotic acid into cytidine nucleotides is markedly activated; the degree of activation is lower in regenerating livers of alpha-HCH treated animals. The changes in the level of cytochrome P-450 vary inversely with the changes in the utilization of [2-14C] orotic acid for the synthesis of cytidine nucleotides. The activity of cytidine triphosphate synthetase of liver cytosol increases shortly after the administration of alpha-HCH; uridine-cytidine kinase is enhanced in the later stages of the drug action. Within 15-45 min after the administration of alpha-HCH the uptake of [U-14 C] cytidine into the liver and its incorporation into RNA cytosine are increased. After the administration of the drug the uptake of [2-14 C] uridine and its incorporation into RNA uracil is also enhanced whereas its utilization for the synthesis of cytidine nucleotides of the acid-soluble extract as well as for the RNA cytosine are suppressed.


[The major OLA histocompatibility complex of sheep. Recent results in the genetic study of 11 factors previously defined by microlymphocytotoxicity]. The genetic study of 11 lymphocyte factors previously described in sheep was continued. 9 factors are the products of alleles at two closely linked loci OLA-A and B. The last two factors are the products of two genes at two distinct loci OL-X and OL-Z. Recombinations between the two closely linked loci are very rare (0.6%); but they are frequent between these two loci and OL-X (26%). The OL-Z locus seems to be independent of the two linked loci OLA-A and B. The allele frequencies at the four loci are given.


Cellular histamine release, specific and total serum IgE levels in hay fever patients and controls. The amount of histamine released from blood leucocytes by allergen, the amount of allergen required to release 50% histamine and the total IgE and IgE specific for Dactylis glomerata are compared in nine hay fever patients and five control individuals. The variation with time of total IgE, of specific IgEe and of the percentage of histamine released at a fixed allergen concentration, corresponding to the cellular sensitivity, is shown for one control individual with positive in vitro and skin tests, but without clinical symptoms. For four patients with a low total IgE level the evolution of these parameters during a 1 year treatment period is presented.


Silica gel radioimmunoassay for myelin basic protein. 1. A radioimmunoassay which relies upon the capacity of silica gel to absorb free human basic protein of myelin (BP) is described. 2. Reagents involved in the assay include unlabelled BP, I-BP, and rabbit anti-BP. The assay is performed in excess normal rabbit serum, and uses 3-4% silica gel for absorption of free I-BP. 3. The assay is simple, reproducible and not particularly time-consuming. It is capable of detecting BP at a level of 1-10 or more nanograms in samples containing varying amounts of serum. The assay is thus highly suitable as a routine test in clinical laboratories interested in detecting BP in cerebrospinal fluid in patients with suspected or proven demyelination.


Isoelectric focussing of proteins on cellulose acetate gel membranes. A method is described for analytical isoelectric focussing of proteins on cellulose acetate gel strips pretreated with boron trifluoride in methanol. It is satisfactory for separations requiring narrow pH gradients as well as for those requiring wide gradients. As well as conventional protein staining methods, the use of immunological reagents is described to identify directly specific proteins in a complex separation. The method is extremely quick and easy to perform and requires only low voltage electrophoresis equipment.


An investigation into creatine kinase and other plasma enzymes in thyroid disorders. The relationship between plasma protein bound iodine (PBI) level and creatine kinase (CK) activity was investigated in 143 males and 528 females suspected of various thyroid disorders; there was significant negative correlation between low PBI level and raised CK activity. CK, aldolase, lactate dehydrogenase (LD), aspartate transaminase (AST), and alanine transaminase (ALT) activities were determined in plasma from patients with reduced PBI levels; apart from CK, LD was the only enzyme increased in an appreciable number of cases. A further series of specimens was collected from 66 patients with low PBI levels and the CK isoenzymes investigated. In all of these MM was the main form present; a trace of MB was found in 6. These findings do not explain the elevation of CK in hypothyroidism which may be a non-specific effect.


Cytostasis against self-antigens.

Cytostasis against self-antigens. Effector cells from mice of the C57Bl/10 congenic series, alloimmunized against K end- or D end-identical I region incompatible lymphocytes, were cytostatic for tumour and macrophage targets bearing syngeneic K-end and D-end antigens. If incompatibility during immunization was restricted to the I-B subregion only, cytostasis was directed against K end syngeneic antigens; further incompatibility in the I region was required to obtain cytostasis against D-end self-antigens. Cytostasis against self-antigens was also obtained after immunization with syngeneic tumour cells; in one instance this was directed against syngeneic K-end antigens only. The implications of these findings in relation to neoplastic change and autoimmune damage are discussed.


Evidence for a change in the expression of beta2-microglobulin-assoicated membrane structures on leukaemic human cells. Cell-associated and serum beta2-microglobulin was estimated in seven patients with chronic lymphocytic leukaemia. The amount of cell-associated beta2-microglobulin was significantly reduced (P less than 0.01), due to a decrease in the fraction of beta2-microglobulin that passes unretarded through a concanavalin A affinity column (presumably non-HLA-associated beta2-microglobulin). Serum concentrations of beta2-microglobulin were increased, but no correlation was found between the decrease in cell-associated beta2-microglobulin and the increase in serum beta2-microglobulin. All of the beta2-microglobulin from leukaemic serum was eluted corresponding to a molecular weight of 11,800 and none of it was retarded on a concanavalin A affinity column. The decrease in cell-associated beta2-microglobulin might reflect a change in the qualitative or quantitative expression of beta2-microglobulin-associated membrane structures on the leukaemic cells, perhaps conferring resistance to the cells against hypothetical immunological host defence mechanisms.


A new chromosome type replacing the double minutes in a mouse tumor. For several years the SEWA mouse ascites tumor has been a carrier of double minute chromosomes (DMs), some 90% of its cells containing from one to several hundred DMs. In one specific subline of this tumor, the cells with DMs had decreased in frequency to less than 5% of the cells. At the same time, the stemline chromosome number had increased from 43 to around 50. This was due to the presence, in addition to the ordinary telocentric chromosomes, of a varying number of medium-sized metacentrics. The fact that these chromosomes deviated from ordinary mouse chromosomes in special features, such as median centromeric position, early DNA replication, and complete lack of centromeric heterochromatin, indicates that they represent a new type of chromosome. Their striking agreement with the DMs in many properties makes it tempting to associate their origin with the disappearance of the DMs.


The nature of the Ag-staining of nucleolus organizer regions. Electron- and light-microscopic studies on human cells in interphase, mitosis, and meiosis. Electron micrographs reveal that the Ag-stainable substance is located on the outside of NOR's or around them but not in the chromosomes themselves. In association figures, the Ag-positive material lies between the acrocentric chromosomes. Light-microscopic studies show that the Ag stainability of the nucleolus in interphase is correlated with the function of the NOR, as seen from inactive and activated lymphocytes. Much more Ag-positive material is seen in prophase than in meta- and anaphase. It starts to increase again in late telophase. In male meiosis the NOR's remain Ag-positive until pachytene. First and second metaphase figures are negative. Experiments using RNase, TCA, and trypsin indicate that the Ag-stainable substance is an acidic protein. The precipitation of Ag granules in interphase nuclei seen in the electron microscope is greatest over the fibrillar component of the nucleolus. The most likely interpretation is that the Ag-stainable material is a component of ribonucleic protein accumulating around active NOR's. In mitosis some of this material remains at the NOR's. In first meiosis it is completely removed before diakinesis.


Testicular cells lysostripped of H-Y antigen organize ovarian follicle-like aggregates. A suspension of free testicular cells were obtained by mild trypsin treatment from newborn BALB/c testes, and their plasma membrane H-Y antigen sites were blocked (lysostripped) by an excess of H-Y antibody of proven specificity and potency (45 min in ice). Upon 16 h of the Moscona-type rotation culture, these treated testicular cells yielded primarily spherical aggregates, more than half of which demonstrated a strong resemblance to ovarian follicles. The resemblance was particularly striking between the smallest testicular folliculoids and primordial ovarian follicles that abound in the newborn female gonad. Under the same condition, control serum-treated testicular cells primarily yielded cylindrical tubular structures that can be very long. Over a critical range, concentrations of H-Y antibody apparently influenced the frequency of testicular folliculoid formation. The above directly supports the proposed testis-organizing function of H-Y antigen and is certainly compatible with the genetic situation encountered in the wood lemming (Myopus schisticolor), that in the functional absence of H-Y antigen, XY gonadal cells readily organize an ovary.


Nucleolus organizer activity and the origin of Robertsonian translocations. Chromosomes with active nucleolus organizer regions (NOR's) were identified by combined Q-banding (in some cases), and silver staining in mouse cell lines. NOR-bearing chromosomes were overrepresented among the chromosomes involved in Robertsonian translocations in LM(TK-), A9, and RAG cell lines. Usually only one NOR-bearing chromosome was seen in any biarmed chromosome; relatively few contained two NOR-bearing chromosomes. Thus the nucleolus plays an important role, but nucleolar fusion is relatively unimportant, in the origin of Robertsonian translocations in the mouse.


Aneuploidy and ageing: chromosome studies on a random sample of the population using G-banding. Chromosome analysis using G-banding was carried out on cells from 65 males and 102 females of all ages from a random sample of the population. The frequency of aneuploid cells showed a significant increase with age in both sexes, and in females the increase in hypodiploidy and hyperdiploidy was more marked than in males, and involved a high proportion of cells that had lost or gained an X chromosome, 45,X cells being much more common than 47,XXX cells. In females, the occurrence of a "fragment" of an X chromosome also correlated with increasing age, and this "fragment" appears to be an X chromosome that has simply divided prematurely at the centromere. The effects of time in culture and of repeating cultures of blood samples from the same individual on proportions of abnormal cells of various types were also investigated, and the results are discussed in the light of findings from several other "ageing surveys".


Human tumor and rodent-human hybrid cells with an increased number of active human NORs. A human fibrosarcoma line, HT1080-6TG, with a near diploid number of chromosomes, has an average of 7.3 chromosomes with an Ag-stained nucleolus organizer region (NOR). Cells of this line with an increased number of chromosomes have an increased number of Ag-stained NORs. This cell line has been used as the human parent in constructing mouse-human and rat-human hybrids that segregate rodent chromosomes. The hybrid ccell lines, which have 100 or more chromosomes per cell, show a proportionate increase in the number of Ag-stained NORs (means, 11.4--16.8). The frequency of association of acrocentric chromosomes increases in a similar fashion. There is no evidence of inactivation of human NORs in these cells.


Induced Robertsonian fusions and tandem translocations in mammalian cell cultures. Cultures of a cattle cell line and a Peromyscus eremicus cell line recovering from a pulse-treatment with mitomycin C, actinomycin D, 33258 Hoechst, and nitrosoguanidine exhibited translocations between chromosomes at the centromeric regions (Robertsonian fusions) as well as between centromere and telomere and between telomeres (tandem translocations). The frequency of Robertsonian fusions was found to be dose-dependent and duration-dependent with the mitomycin treatment. Biarmed chromosomes resulting from fusions may be monocentric or dicentric. Analyses of clones isolated from treated cells suggested that fused chromosomes may perpetuate in the cell populations.


Radiation induced isostaining: fact or fiction? Two different experiments were set up to induce differential staining in M-1(1) cells or to induce isostaining in M-2 cells after gamma-irradiation of human lymphocytes. Neither of these two unusual staining patterns previously described by Luchnik and Porjadkova (1977) were observed. Possible explanations for this disagreement are discussed.


Immunodiffusion studies on Schistosoma mansoni and its host stage specific antigens. 1. Immunoelectrophoresis cross-reactions between S. mansoni adults and larval stages antigens. IMMUNOELECTROPHORESIS was applied to evaluate the immunologic relationship between the different stages in the life cycle of Schistosoma mansoni (S.m.). The preparation of antigens and antisera was described. Separated female and male adult worms tested seem to be identical. Eggs, miracidia and cercariae from Biomphalaria glabrata and Biomphalaria alexandrina showed varying degree of cross-reactions. The maximal number of pricipitin arcs was observed, when anti-miracidia-serum was checked against all other S.m. antigens.


Immunodiffusion studies on Schistosoma mansoni and its host stage specific antigens. 2. Immunoelectrophoresis cross-reactions between hepato-pancreas of Biomphalaria glabrata and Biomphalaria alexandrina. IMMUNOELECTROPHORESIS was used for studies on immunological cross-reactions between both Schistosoma mansoni (S. m.) intermediate host snails Biomphalaria glabrata and Biomphalaria alexandrina. The preparation of antigens and their corresponding antisera from hepato-pancreas of S. m. infected and non infected snails was described as the homologous reactions of both snails either infected or non infected were rather similar, close immunological relationship between B. glabrata and B. alexandrina could be deduced.


Immunodiffusion studies on Schistosoma mansoni and its intermediate host stage specific antigens. 3. Immunoelectrophoresis cross-reactions between S. mansoni stages and Biomphalaria sp. hepato-pancreas antigens. IMMUNOELECTROPHORESIS was applied for cross-wise checking of the different stages in the life cycle of schistosoma mansoni with extracts of hepato-pancreas from its intermediate host snails Biomphalaria glabrata and Biomphalaria alexandrina. Only few fractions were common to non infected snail organ and parasite development stage, while more crossreactions were detected with infected snail hepato - pancreas.


Enzymatic synthesis of blood-group Lewis-specific glycolipids. A blood-group Lewis precursor glycolipid was isolated from the plasma of a Lewis-negative individual [Le(a--b--)] and treated with fucosyltransferases from human gastric mucosa and GDP-fucose. Subsequently the glycolipid was adsorbed onto Le(a--b--) erythrocytes and the presence of blood-group Lewis antigens was assessed by passive hemagglutination with anti-Lewis sera. It was shown that the precursor glycolipid was enzymatically transformed to blood-group Lewis a (Lea) and Lewis b (Leb) specific glycolipids. Leb-glycolipid was also synthesized by fucosylation of an isolated Lea-glycolipid. Moreover Le(a--b--) erythrocytes were shown to develop Lea and Leb activities when subjected to enzymatic fucosylation, thus showing that Lewis-negative cells carry blood-group Lewis precursor glycolipid on the surface of their membrane. Le(a + b--) erthrocytes, upon enzymatic fucosylation, acquired Leb activity.


Yeast DNA polymerases: antigenic relationship, use of RNA primer and associated exonuclease activity. Highly purified preparation of DNA polymerases A and B from yeast were compared with respect to antigenic relationship, ability to use ribonucleotide primers and associated nuclease activity. The following results were obtained. 1. Antiserum directed against DNA polymerase A inhibits this enzyme but does not interfere with activity of DNA polymerase B or of mitochondrial DNA polymerase, nor does it precipitate the latter two enzymes. 2. DNA polymerase A is capable of using oligo(ribouridylic acid) as a primer for the polymerization of dTMP. This reaction is not catalyzed by polymerase B to any significant extent. 3. Whereas DNA polymerase A is devoid of nuclease activity, DNA polymerase B catalyses an exonucleolytic release of mononucleotide units from the 3' end of polynucleotides. The results of several experiments suggest that this nuclease activity is associated with the DNA polymerase B molecule.


beta-Lapachone, an inhibitor of oncornavirus reverse transcriptase and eukaryotic DNA polymerase-alpha. Inhibitory effect, thiol dependence and specificity. beta-Lapachone is a naturally occuring compound that can be isolated from a number of tropical trees. It is shown to be a potent inhibitor of reverse transcriptase activity from both avian myeloblastosis virus and Rauscher murine leukaemia virus. In addition, it affects eukaryotic DNA-dependent DNA polymerase-alpha activity: 50% inhibition is reached in 60-min incubation time by about 8 micron beta-lapachone. Enzyme activity is inhibited irrespective of the purity of the enzyme used or of the amount or type of template/primer or substrate present. The inhibitory effect of the drug is only observed in the presence of dithiothreitol. The primary site of action of beta-lapachone appears to be the enzyme protein, as is also borne out by the specificity of its action. Eukaryotic DNA-dependent DNA polymerase-beta, prokaryotic DNA-dependent DNA polymerase I, several other nucleic acid polymerases and some completely unrelated enzymes are not affected. Reverse transcriptase and DNA-dependent DNA polymerase-alpha may be in someway related in possessing similarly exposed '--SH structures' in their active sites. beta-lapachone thus affords a novel means of studying such interrelationships and of further characterizing enzymes.


Interdependent extrasystoles in a patient with a biventricular bypass heterotopic cardiac homograft. This report describes the induction of atrial extrasystoles in the donor heart by preceding ventricular extrasystoles in the recipient heart of a patient with a biventricular bypass heterotopic cardiac homograft. Timing of the ventricular extrasystoles of the recipient heart in relationship to the preceding donor heart systole was critical to the production of donor heart atrial extrasystoles. The mechanism is thought to be a mechanical one resulting from an acute pressure rise in the atrium of the donor heart due to increased mitral regurgitation from the recipient heart resulting from the ventricular extrasystoles.


Effect of paramethasone acetate on women with secondary amenorrhea: a preliminary report. Twelve women of normal weight (ages 17 to 36 years) with scanty menstrual bleeding were studied. They had no signs of virilization, gynecologic or endocrine pathology, or past history of hormonal treatment. Five women (group 1) experienced withdrawal bleeding after a 3-day course of chlormadinone acetate (2 mg/day) and the other seven did not (group 2). Daily venous blood samples were obtained 10 to 15 days afterward for 5 consecutive days of no treatment (control period) and during the next 5 days while the women received paramethasone acetate (PA), 2 mg/day (treatment period). In each plasma sample the concentrations of 17beta-estradiol (E2) and luteinizing hormone (LH; LER-907) were determined. The mean plasma E2 levels in group 1 were 35 +/- 8 and 86 +/- 10 pg/ml during the control and treatment periods, respectively (P less than 0.001), and the mean plasma LH levels were 28 +/- 6 and 94 +/- 34 ng/ml, respectively (P less than 0.001). No significant changes in plasma E2 and LH levels were observed in group 2 in either period. During the control period, the plasma E2 level in group 2 (14 +/- 2 pg/ml) was lower than that in group 1 (P less than 0.01); however, plasma LH levels were similar in both groups. The administration of PA for 5 months induced monthly ovulation in group 1 but not in group 2. These data suggest that the best results are obtained in women with optimal pretreatment levels of plasma E2.


Demonstration of a chorionic gonadotropin-like substance in rabbit morulae. Rabbit morulae were treated with specific indirect immunofluorescence and immunoperoxidase histochemical techniques. The first antibody in both systems was rabbit anti-human chorionic gonadotropin (hCG) beta-subunit. Negative controls revealed a complete absence of reaction. All morulae incubated with the double-antibody system showed a positive reaction. Rabbit morulae thus present a substance with antigenic determinants similar to the beta-subunit of hCG. The physiologic role of this substance is unknown.


Type I diabetes mellitus.

Type I diabetes mellitus. The major genetic susceptibility to insulin dependent (Type 1) diabetes is determined by genes in the HLA chromosomal region. An increased relative risk for developing the disease is observed in subjects who are HLA A1, A2, B8, B18, B15, B40, CW3, Bfs, DW3, DW4, DRW3, DRW4 positive. There is an additive relative risk in subjects who possess two "high risk" HLA B alleles which has an important influence on the prevalence of the disease in sibships and possibly on the concordance rate in diabetic identical twins. There is also suggestive evidence that particular combinations of "high risk" HLA B alleles are associated with increased or persistent antibody production which may reflect enhanced or differential susceptibility. Certain factors (e.g. HLA B7, DW2 and DRW2) are associated with a significantly reduced risk and may exert a "protective" mechanism in Type I diabetes, by linkage disequilibrium with genes which reduce immune responsiveness. The significant increases and decreases in respect of the HLA B antigens are probably secondary to the corresponding HLA D and DRW associations which reflect a stronger linkage disequilibrium between the genes which determine these specificities and the putative genes which control susceptibility. Initial damage to the beta cells probably occurs a considerable time before the onset of symptoms and theoretically modification of the immune response early in the disease process may reduce the rate of beta cell destruction.


Quantitative immunodiffusion assay of serum protein patterns in protein calorie malnutrition (PCM). Serum protein components were evaluated by the specific radial immunodiffusion technique in a number of protein energy malnourished infants and young children, as well as in a group of parallel controls. Results show a drop in both serum prealbumin and albumin in concordance with the severity of the condition in both KWO and marasmus. Serum level of varies as 1-acidic glycoprotein was elevated, being more marked in KWO than in marasmus. Serum oc1-antitrypsin, haemopexin, ceruloplasmin and transferrin were all reduced but to a varying extent in both KWO and marasmus. The levels of serum IgA and IgM were above normal in all disease entities except for a slight decrease of IgA in 2nd grade marasmus. On the other hand, serum IgG was reduced in the majority of cases. The ratio of serum cerutoplasmin to transferrin in KWO was found to be of diagnostic value in different grades of the disease.


A new method for treating prostatic hypertrophy. Since 1969, when I did the first transurethral prostatic incision on a 51-year old man with bladder neck obstruction, 245 patients have undergone this procedure; 220 have been followed for 3 months to 8 years. Almost one-half of these patients were between 70 and 90 years old. Comparison of the first 220 patients who had transurethral incision with 220 patients who had transurethral prostatic resection showed that only two who underwent incision required subsequent resection, while 8 patients in the resection group had to have repeat resection. Contracture of the bladder neck and urinary incontinence did not occur with transurethral incision, and the incidence of epididymo-orchitis and stricture is lower with this method than with resection.


Serum and tumour ferritins in primary liver cancer. Serum ferritin concentrations were found to be raised, often considerably, in 58 of 76 black patients with primary liver cancer (PLC). No correlation could be demonstrated between the serum ferritin concentration and several other measurements, including the following: hepatic iron stores measured chemically, the size of the tumour, serum transaminase values, and the presence or absence of cirrhosis in the non-tumorous liver. There was, however, a negative correlation between serum ferritin and alpha-foetoprotein concentrations. Ferritin was purified from PLC tissue obtained from three patients at necropsy and the distribution of isoferritins was determined by isoelectric focusing. Acidic isoferritins similar to those previously found in PLC tissue were obtained. Their acidic nature was confirmed chromatographically using DEAE cellulose. Because the serum ferritin in patients with PLC probably consists of a mixture of normal and acidic isoferritins, it is likely that the serum assay used in the present study underestimated the actual concentrations present. With the development of an assay which utlises a specific antibody against acidic PLC isoferritins, serum ferritin may prove to be a second marker for PLC.


Silver staining studies on the short arm variant of human chromosome 17. The absence of silver grain precipitation on human chromosome 17, consistently observed in four patients displaying the 17p variant, supports the hypothesis that the 17p variant is a structural heteromorphism rather than translocated satellite material.


Immunosuppressive activity of isolated rat and human fetoproteins. Alpha-fetoprotein (AFP) was purified from rat hepatoma sera and pooled human cord sera. Both AFP-rich fractions prepared by consecutive ion exchange and gel filtration chromatography and pure rat AFP prepared by liquid phase immunoabsorption lacked in vitro suppressive activity. Human AFP purified by affinity chromatography was suppressive, but so was similarly purified human cord albumin. Alteration of ionic conditions was shown to affect the activity of both human AFP and cord albumin. The primary mixed lymphocyte response, the generation of the secondary response, and the memory cell, per se, were all found to be sensitive to active human AFP under the appropriate culture conditions.


Human alpha-fetoprotein: immunosuppressive activity and microheterogeneity. Purified human alpha-fetoprotein (HAFP) from five patients with hepatoma, one with gastric carcinoma, one with an embryonal cell tumor, and from fetal liver has demonstrated immunosuppressive potencies in vitro which vary over three orders of magnitude. A reversible association of HAFP with the cell surface and a predominant effect on T cells are suggested. No evidence of complex formation between HAFP and mitogen has been found. The microheterogeneity of HAFP has been detailed with crossed immunoelectrophoresis and isoelectric focusing in polyacrylamide gels containing 8M urea, and the immunosuppressive potency of HAFP isolated from a given source can be correlated with the proportion of certain HAFP species contained in it.


Immunosuppressive characteristics of human AFP: effect on tests of cell mediated immunity and induction of human suppressor cells. Murine AFP has been reported to be immunosuppressive in a variety of systems. However, the extent and degree of inhibition has varied in different species and laboratories. Therefore, we have examined the potential suppressive effect of purified human AFP on several in vitro tests of cellular immunity and the potential mechanism of its action. AFP purified from fetal and liver cancer sera significantly inhibited mitogen and antigen-induced proliferative responses but had no effect on lymphocyte E rosetting, MIF production or mitogen induced T cell cytotoxicity to Chang target cells. Purified human AFP induced human suppressor cell activity, capable of suppressing a one-way mixed lymphocyte reaction (MLC). In contrast to Con A induced suppressor cells, AFP induced suppressor cell activity was overcome by mitogen augmentation of the proliferative response in MLC. These data suggest that the inhibition of lymphocyte proliferation by human AFP may be mediated by the induction of a subpopulation of human suppressor cells. Furthermore, mitogen induced cell mediated cytotoxicity was partially inhibited by primary liver cancer serum and completely inhibited by newborn cord serum, in contrast to purified fetal or tumor AFP which had no effect. These data suggest that there are other immunosuppressive factors in fetal and tumor serum which require further characterization. These other serum factors may be responsible for some of the immunosuppressive effects attributed to AFP. Although AFP is unlikely to play a major immunosuppressive role physiologically in vivo, its selective effect on proliferative responses, apparently mediated by suppressor cells, may prove to be a useful pharmacologic probe of the mechanism of these in vitro lymphocyte responses and biological interactions.


A population of human cord blood mononuclear cells with surface alpha fetoprotein. Suspensions of mononuclear cells from adult peripheral blood (PBL) and mononuclear cells from cord blood (CBL) were examined for the presence of surface alpha-fetoprotein (AFP) using a fluoresceinated F(ab')2 fragment of rabbit IgG anti-human AFP. The mean proportion of CBL with AFP was increased (10%) when compared with PBL (1%) although some CBL specimens did not demonstrate such an increase (range 0--15%). The presence of AFP on CBL could be either due to cytophilic AFP attached to a unique surface receptor or intrinsic AFP synthesis. The following observations could not distinguish between these two possibilities: (1) After treatment with trypsin, only minor reappearance of surface AFP could be observed in AFP-free medium in contrast to the larger numbers observed in medium containing AFP. Such selective reappearance depending on the media could be related to either cytophilic attachment of heterologous or homologous AFP or preferential stimulation of intrinsic AFP synthesis. (2) The reappearance of AFP positive CBL following trypsin treatment and incubation in media with or without AFP containing sera was inhibited by cyclohexamide. Such inhibition could be due to inhibition of synthesis of an AFP surface receptor or intrinsic AFP. (3) The shedding of surface AFP observed at 2--4 degrees C could be due to release of exogenous cytophilic AFP or the continued "turnover" of intrinsic AFP without concomitant AFP synthesis due to the cold temperature. Finally, the removal of AFP positive cells via selective depletion of B cells using bead columns coated with IgG-anti-IgG and the absence of depletion of AFP positive cells after successive gradient centrifugation of E-rosettes and cells with IgG-Fc receptors are consistent with the identity of AFP positive CBL as cells without IgG-Fc receptors or lymphocytes without conventional T-cell markers as defined by E-rosettes.


Immunochemical properties of alpha-fetoprotein (AFP) and antibodies to autologous AFP. Alpha-fetoprotein (AFP) is a strong immunogen when injected into another species, but does not elicit an immune response in the species of origin of the AFP. AFPs from different mammalian species are crossreactive. AFP and albumin show sequence homology. They do not crossreact in their native state, but their unfolded (reduced and carboxamidomethylated) forms are immunologically crossreactive. Antibodies to autologous AFP can be induced in animals by immunization with heterologous or modified homologous AFP. These antibodies eliminate the AFP normally measurable in the serum and delay the elevation of serum AFP caused by hepatomas, but do not protect against such tumors.


[Clinical experiences with a 6% hydroxyethyl starch 40000 solution; metabolic parameters and tolerance]. A 6% hycroxyethyl starch solution with a mean molecular weight of 40 000 and a degree of substitution of 0.5--0.55 was tested under controlled conditions at 79 clinics in West Germany. The following parameters were evaluated: tolerance, circulation effectiveness, influence on serum electrolytes, hemoglobin and hematocrit. A total of 779 patients received either 6% HES 40 000 alone, 6% HES 40 000 and electrolyte solution or 6% HES 40 000 solution and other volume effective plasma substitutes for treatment of hypovolemia. The results show that stable circulation can be achieved in 4 to 6 hours during narcosis as well as in the subsequent phase with a sufficient supply of fluid through 6% HES 40 000 solution. One case of flush and a slight chill were reported as side effects related to the hydroxyethyl starch.


Immune competence in a patient with Hodgkin's disease and relapsing toxoplasmosis. A 40 year old woman with Hodgkin's disease twice developed signs of encephalitis while being treated with prednisone and cyclophosphamide for 10 months. Since on both occasions her Toxoplasma dye test titer was 1 : 8000 or higher, she was treated on suspicion of toxoplasmosis with sulfadizine and pyrimethamine. Her tumor therapy was changed to bleomycin with lower doses of prednisone for 12 months. After death from central pontine myelinolysis, Toxoplasma and cytomegalovirus could be isolated, but no lesions attributable to these infectious agents were present. Maintenance of the patient's immune competence suggested an inquiry into the effects of the chemotherapeutic agents and of tumor infiltration for their respective interference with immunity. Using hamsters with chronic latent toxoplasmosis, it was found that both cortisone and cyclophosphamide caused recrudescence of chronic inapparent infection, that vinblastine and bleomycin interfered only slightly with the development of immunity, whereas in infiltrating lymphoma permitted immunity to develop normally. It is concluded that greater attention should be directed to the immunosuppressive effects of tumor treatment. By choice of an effective tumor therapy which is least immunosuppressive, and if necessary under cover of antimicrobial therapy, a patient with Hodgkin's disease can be aided in developing immunities which he may subsequently be able to maintain.


Therapeutic and prognostic aspects of cancer of the prostate detected incidentally in the course of adenoma enucleation. Though cancer of the prostate is generally regarded as a fairly benign tumour with long survival, many patients die of it. The sovereignty of hormone therapy seems questionable in the light of recent experience. A selected patient material with early stage cancer of the prostate--incidental findings at the Freyer-Hryntschak operation for bladder neck adenoma--is presented for the illustration of the prognostic aspects of this tumour. Androgen control therapy was given postoperatively to every patient.


Ultrastructural carbohydrate cytochemistry of gastric epithelium. On examination with ultrastructural methods for visualizing the vicinal glycols and acid groups of complex carbohydrates, the most superficial surface epithelium of the rat gastric corpus displayed biphasic mucous droplets consisting of a cortex of hexose-rich (i.e. periodate-reactive) neutral mucosubstance and an uncharacterized denser core plus monophasic droplets with the neutral mucosubstance. In many surface epithelial cells of the foveolae, the biphasic and monophasic droplets with the neutral mucosubstance intermingled in varying proportions with monophasic droplets showing uniform periodate reactivity, a variable degree of dialyzed iron-binding--demonstrative of acidic glycoconjugate, and high iron--diamine affinity--demonstrative of sulphomucin. Deep foveolar epithelium displayed only monophasic droplets, most of which contained acidic periodate-reactive complex carbohydrate. Underlying cells, designated isthmus cells, exhibited monophasic or occasional biphasic granules containing sulphated, hexose-rich mucosubstance. Nascent droplets or granules near the Golgi zone differed from the mature organelles in the distribution of the glycoconjugate. Mucous neck cells occupied a deeper stratum and displayed a uniform population of monophasic mucous droplets with a loose meshwork of neutral mucosubstance. Techniques for demonstrating hexoses ultrastructurally stained all Golgi cisternae in the mucigenic epithelium, showing increasing reactivity toward the maturing face. Distinctive cisternae with moderate reactivity in the Golgi complex of isthmus cells were interpreted as GERL. Acidic mucosubstances were visualized only in the inner, mature cisternae of the Golgi complex of cells storing acidic glycoconjugates, and not in cisternae interpretable as GERL. The apical plasmalemma of isthmus cells uniquely exhibited abundant sulphated glycoconjugate and that of parietal cells revealed a less prominent, periodic neutral mucosubstance. Lateral and basal plasmalemmae varied from unstained to slightly reactive; basement membranes showed moderate reactivity with methods for visualizing complex carbohydrates. Abundance of glycogen further characterized surface epithelial cells of the corpus and of some parietal cells.


Alkaline phosphatase possessing alkaline phosphodiesterase activity and other phosphodiesterases in Bacillus subtilis. In Bacillus subtilis Marburg strain, single-point mutations in the phoP locus brought about simultaneous losses of the major activities of alkaline phosphatase (APase) and alkaline phosphodiesterase (APDase). Revertants recovered the two activities. APases with APDase activity were purified from the membrane fraction of B. subtilis 6160-BC6 and from the culture fluid of an APase-secreting B. subtilis mutant strain, RAN 1. In addition to these major APases with APDase activity, at least two kinds of phosphodiesterase (PDase) without phosphatase activity were found in the cytoplasmic supernatants of RAN 1 and an APase-less B. subtilis mutant strain, SP25. Another minor APase with a molecular weight of about 80,000, which had almost no PDase activity, was isolated from the membrane fraction of strain 6160-BC6. Enzyme distribution in subcellular fractions from various strains cultured in high- and low-phosphate media was analyzed. The PDases did not cross-react with rabbit antiserum against the RAN 1 APase with APDase activity. The main component of the PDases had a molecular weight of about 80,000 and was most active at pH 8.0. These results suggest that APase with APDase activity is different from PDases detected in cytoplasmic supernatants and that phoP is the structural gene for the phosphate-repressible APase with APDase activity.


Purification and properties of 2-hydroxy-6-oxo-2,4-heptadienoate hydrolase from two strains of Pseudomonas putida. Growth on phenol of two strains of Pseudomonas putida biotype A, NCIB 10015 and NCIB 9865, elicits the synthesis of an enzyme that hydrolyzes 2-hydroxy-6-oxo-2,4-heptadienoate to 2-oxopent-4-enoate. The purified enzyme from Pseudomonas NCIB 10015 has a molecular weight of 118,000 and dissociates in sodium dodecyl sulfate to a species of molecular weight 27,700; the enzyme from Pseudomonas NCIB 9865 has a molecular weight of 100,000 and dissociates to a species of 25,000 molecular weight. The hydrolases from both strains have similar Km values, pH optima, and thermal labilities and attack the same range of substrates. Neither hydrolase was stimulated by Mg2+ or Mn2+, and both were inhibited by p-chloromercuribenzoate and iodoacetamide. Immunodiffusion studies with the purified enzymes and antibodies formed against them show some cross-reaction of Pseudomonas NCIB 9865 enzymes with antibodies to Pseudomonas NCIB 10015, but not vice versa.


Platelet-foreign surface interactions: release of granule constituents from adherent platelets. The interaction of suspensions of washed pig platelets with nine artificial surfaces (glass, polystyrene, three segmented polyurethanes, and surfaces formed by coating glass with albumin, fibrinogen, gamma-globulin, and collagen) is reported. Platelet adhesion and release from adherent platelets were measured via labeling with 51Cr and 14C-serotonin. The apparatus was a couette flow device allowing observation of surface-platelet effects uncomplicated by transport or flow effects. Using a two-level factorial design the effects of albumin, fibrinogen, red cells, and platelet count on adhesion and release were estimated for each surface. Comparison of the various surfaces showed that collagen and gamma-globulin are the most reactive (mean adhesion, 34 platelets/100 micrometer2, mean release 50% of granule contents). The other surfaces showed lower levels of release (approximately 25%), indistinguishable one from another. The adhesion levels of two hydrophilic polyurethanes and albumin were low, while those of the remaining surfaces were moderate. The effect of albumin was to reduce adhesion for the "moderate" group of surfaces. Fibrinogen increased adhesion to nonprotein surfaces and decreased release for collagen and gamma-globulin surfaces. High platelet count increased adhesion to fibrinogen, gamma-globulin, and collagen surfaces. Red cells increased adhesion to all surfaces and increased release for collagen and gamma-globulin.


Adsorption of plasma proteins on hydrophobic surfaces. II. Fibrinogen and fibrinogen-containing protein mixtures. The adsorption of fibrinogen from single solution and from mixed solution with albumin and gamma-globulin has been followed at the air-buffer, isooctane-buffer, and methylene iodide-buffer interfaces by the pendant drop technique. Fibrinogen is shown to form substantial coherent films on isooctane and methylene iodide, suggesting considerable unfolding and lateral association. From this, a novel hypothesis has been proposed to account for the dominance of fibrinogen adsorption from mixed solutions on certain hydrophobic surfaces, in which the spreading pressure of rapidly unfolding fibrinogen molecules is sufficient to desorb other plasma proteins.


Changes in proteoglycan and collagen in cartilage in rheumatoid arthritis. Articular cartilage from thirteen patients with rheumatoid arthritis was studied by both light and electron microscopy using recently developed staining techniques for proteoglycan. Normally proteoglycan is concentrated about active chondrocytes, but in the cartilage from these patients it was found to be diminished about most of the chondrocytes while it was increased around a few. Fragmentation of collagen and phagocytosis of the fragments by chondrocytes were also noted. This dissolution of collagen was associated with loss of proteoglycan in the matrix. These findings are consistent with the variable nature of the rheumatoid destructive process and appear to be morphological reflections of biochemical changes which formerly were thought to occur in this disease only at the cartilage-pannus junction.


The capping of lymphocytes and other cells, studied by an improved method for immunofluorescence staining of frozen sections. Experiments have been carried out on the capping by lectins and antibodies of surface receptors of mouse splenic T and B lymphocytes and other cells, in which the surface distribution of the lectin or antibody, and the intracellular distribution of myosin or actin, were determined on the same cells by a double fluorescence technique. For this purpose, a general method for intracellular staining was developed which is intended to preserve sensitive antigens and fragile ultrastructural elements. The method involves mild formaldehyde fixation of the cells or tissues, infusion with concentrated sucrose, rapid freezing, and the preparation of frozen sections thinner than 1 micrometer thickness. The immunofluorescent or other appropriate fluorescent reagents are then applied to the thawed section. In the present experiments, intracellular actin was detected using a fluorescent staining method based on the interaction of F-actin with heavy meromyosin, while intracellular myosin was detected by an indirect immunofluorescence procedure. Our findings were that the formation of a cap by each of the lectins or antibody reagents was always accompanied by a concentration of myosin and actin directly under the cap. These and other results suggest that capping is an active process in which actin and myosin participate directly in the formation of all caps. This proposal carries important new implications for the molecular mechanism of capping.


[Efficacy of biliary by-pass operations in carcinoma of the head of the pancreas with jaundice (author's transl)]. In the present state of detection and treatment of carcinoma of the head of the pancreas, only palliative surgery is available. The debate concerning the type of bilio-digestive by-pass is far from closed. Although by-passes using the gall bladder remain rare in France, above all reserved for advanced cancer, we prefer to use the common bile duct. The choice of the by-pass depends mainly on the local conditions and on the general state of the patient. Side-to-side choledoco-duodenal anastomosis, provided the technic is perfect and there is no duodenal obstruction, remains the simplest and therefore the most commonly used by-pass. However in cases of large pancreatic carcinomas modifying the relations between the duodenum and the bile duct or stenosing the duodenum, or as a routine whenever possible, it seems to us advisable to undertake a choledoco-jejunostomy on a Y loop (the raised loop is also used for a gastrojejunostomy), and this seems to give the most rapid recovery from the jaundice, a low mortality and ensure the longest survival.


Mycobacterium chelonei iatrogenic infections. We report on two outbreaks of Mycobacterium chelonei subsp. abscessus cutaneous infections, which occurred between June 1974 and April 1975 in a series of 24 patients (15 studied bacteriologically) subjected to venous stripping for varicose veins. The source of infection was the aqueous solution of merbromin used in presurgical care.


Alkaline phosphatase and peroxidase for double immunoenzymatic labelling of cellular constituents. The use of alkaline phosphatase in an immunoenzymatic procedure for the localisation of antigens in paraffin sections or cell smears is described. The results of this method, when applied to the detection of immunoglobulins, lysozyme, or lactoferrin, were comparable in intensity and clarity to those obtained with the PAP immunoperoxidase procedure. Furthermore, double immunoenzymatic labelling (with alkaline phosphatase and peroxidase) of two cellular constituents in a tissue section is possible, the brown peroxidase reaction product contrasting well with the blue alkaline phosphatase product. Since the two antibody 'sandwiches' are applied simultaneously rather than sequentially the total duration of this double immunostaining procedure is only a few minutes longer than that required for detection of a single antigen. It was also found that the unlabelled antibody immunohistological procedure (whether used in conjunction with alkaline phosphatase or peroxidase) can be shortened without loss of sensitivity by carrying out two of the incubation steps simultaneously.


IgM-class rheumatoid factor interference in the solid-phase radioimmunoassay of rubella-specific IgM antibodies. The interference of IgM-class rheumatoid factor (RF) in the solid-phase radioimmunoassay (RIA) of rubella virus IgM antibodies was studied. Acute rubella infections did not significantly activate RF. False-positive rubella antibody results were obtained, however, when patients with raised RF levels were tested. If a low rubella IgG antibody titre was present, a high level of RF was required to cause a false-positive IgM result; conversely, in sera with high IgG titres, only a low level of RF was required for interference. Although the false-positive IgM titres obtained were generally low, thet did show a positive correlation to both RF levels and rubella IgG titres. False-positive results were successfully avoided by removing the RF by absorption with heat-aggregated human gamma globulin. The absorption procedure did not affect true rubella IgM antibody titres.


A morphological study of the axons and recurrent axon collaterals of cat sciatic alpha-motoneurons after intracellular staining with horseradish peroxidase. Utilizing the centrifugal neuronal transport of intracellularly injected horseradish peroxidase (HRP), we have performed a light microscopic (LM) investigation of the intramedullary parts of the axons and axon collaterals of sciatic alpha-motoneurons in the adult cat. The intramedullary parts of the alpha-motor axons had comparatively short internodes (down to 75 microns) and were thinner than reported in earlier studies on the ventral root. Positive correlations were obtained when relating nodal diameters (2.8-7.8 micron) or the mean diameters of the motor axons in the white matter (4.4-9.0 micron) to the diameters of the initial axonal segments (2.3-4.9 micron). Eighty percent of the motor axons gave off one to five collaterals. There was no correlation between the numbers of collaterals and the lengths of the parent motor axons in the gray matter. The branching patterns of the axon collaterals showed considerable variation and the number of end branches from a single collateral ranged between 1 and 39. The rostro-caudal distribution of the collateral end branches was arranged symmetrically within a narrow space (+/- 300 micron) around the origins of the first order collaterals. Outbulgings of the motor axon collaterals, interpreted as synaptic terminals, were found along (59%) or at the ends (41%) of the collateral branches, and were located 200-700 micron away from the origin of the first order collateral. No characteristic LM feature of the outbulgings was distinguished.


Demonstration of specific storage material within cutaneous nerves in metachromatic leukodystrophy. Biopsies of clinically unaltered skin from a 5-year-old girl with metachromatic leukodystrophy were studied by light, fluorescence and analytical electron microscopy. Investigation of hematoxylin-eosin-stained sections revealed no pathologic changes, but a brown metachromatic material was found within cutaneous nerves after acetic acid cresyl violet staining of frozen sections. In semithin Epon sections small dermal nerve fascicles contained endoneural deposits, which proved to correspond with the typical prismatic, dense or Tuffstein bodies described in other organs of patients suffering from MLD. The inclusions were located mainly within Schwann cells and exhibited a bright orange fluorescence after trypaflavine-phosphotungstic acid-treatment as well as a high electron density in serial ultrathin sections. Energy dispersive X-ray microanalysis revealed a high sulfur content in the membrane bound granules. These findings demonstrate the presence of the specific storage material also within cutaneous nerves in MLD and thus suggest skin biopsies as an addiitonal and simple diagnostic acid in this disease.


Radioimmunoassay of ovine alpha-fetoprotein. Highly purified ovine alpha-fetoprotein (AFP) was used both for radioisotope labelling and as the reference standard in the double antibody radioimmunoassay of ovine AFP. The sensitivity of the assay is 2 ng/ml which is about 8000 times more sensitive than radialimmunodiffusion assay. The assay is of sufficient sensitivity to quantitate AFP in normal adult sheep serum, pregnancy serum, amniotic fluid and fetal lamb serum.


Primary immune response of guinea pig spleen cells in vitro. Spleen cells of the primed guinea pig, when cultured alone, failed to make a primary immune response against heterologous erythrocytes. When supplemented with lymphocytes obtained from mineral oil induced peritoneal exudate (PEL), a primary immune response against sheep, horse and chicken RBC was obtained. The kinetics, dose response and requirement of PELs for this response are reported.


Measurement of antibody and antigen concentrations with a sodium sulphate glutaraldehyde technique. Gamma-globulins can be crosslinked at low concentrations by simultaneous treatment with sodium sulphate and glutaraldehyde (SSG-method). This precipitate can be used for the assay of antibody or antigens. Examples are given for the measurement of (1) anti-dinitrophenyl antibody by reaction of the precipitate with 125I-dinitrophenylovalbumin, (2) IgA by inhibition of an anti-IgA precipitate with an 125I-labelled IgA myeloma protein, (3) idiotypic antibody by inhibition of the binding of a 125I-labelled myeloma protein by an anti-idiotype precipitate.


A cellulose acetate immunofixation technique. A method of immunofixation using CAM as the only substrate is described. It consists of separating the proteins by conventional electrophoresis followed by immersion for a few minutes only in appropriate specific antiserum suitably diluted with barbitone buffer containing polyethylene glycol (PEG). This is followed by washing and staining by Ponceau S or Nigrosin. The advantages of the method used are speed, sensitivity and economy, no expensive equipment or reagents being required. The method has proved helpful in the identification and classification of minor paraprotein bands and in the analysis of multiclonal paraproteinaemias. The technique can be applied as well to other proteins of biological fluids, e.g. phenotyping.


TB mice. A model of high and long lasting reaginic responders. Reaginic (IgE) antibodies are normally difficult to induce in laboratory rodents. The TB inbred mice constitute an exceptionally suitable model for the study of reaginic responses because of their ability to mount a strong and long lasting (over 4 months) IgE antibody response to hapten carrier conjugates, such as DNP3-OVO, administered in the presence of adjuvant. The maximum individual PCA titres determined for the sera of 8 animals after primary sensitization were in the range of 2,048 to 65,536 (geometric mean: 9,733) and after secondary sensitization the maximum individual PCA titres augmented to a level of 8,192 to 131,072 (geometric mean: 50,490).


An immunosuppressive factor in serum of rabbits lethally infected with the herpesvirus of bovine malignant catarrhal fever. Virus-neutralizing antibody does not protect animals challenged with the herpesvirus of bovine malignant catarrh; therefore, other parameters of the immune response were investigated. In vitro transformation of peripheral blood lymphocytes with mitogens and specific antigens was monitored throughout the course of disease in lethally infected New Zealand white rabbits. The whole-blood culture method was used. Transformation with mitogens was normal throughout the incubation period but fell precipitously from the day of disease onset until death. Specific stimulation with viral antigens was not detected at any time following challenge. Purified and washed lymphocytes, however, reesponded normally to mitogens and were transformed in the presence of viral antigens. Acute-phase serum from rabbits inhibited transformation of normal rabbit lymphocytes, but this inhibition was reversible by washing. It is proposed that the development of a serum suppressive factor(s) is important in the determination of whether an animal is capable of controlling infection with this virus.


Immunologic cross-reactivity between penicillins and cephalosporins: a review. Several approaches have been undertaken in the study of possible immunologic cross-reactivity between cephalosporins and penicillins. Although the chemical structures of these compounds are similar in several respects, there are distinct differences in their degradation and transformation. Various degrees of cross-reactivity of antibodies produced in response to administration of these drugs have been demonstrated both with test systems that measure IgG and IgM antibodies and with those that measure IgE antibodies. The clinical significance of immune responses to cephalosporins is best understood in regard to immunohematologic abnormalities: positive direct antiglobulin (Coombs') tests occur in only approximately 3% of patients receiving cephalosporins; however, several cases of cephalosporin-induced immune hemolytic anemia have been reported. Clinical studies of the cephalosporins indicated that patients with a history of penicillin allergy have increased incidence of reactivity to cephalosporins, but it is impossible to determine to what extent this finding is due to immunologic cross-reactivity because penicillin-allergic patients have an increased incidence of hypersensitivity reactions to drugs immunologically unrelated to penicillins. In addition, there is evidence of specific immune response to cephalosporins that indicates independently acquired hypersensitivity rather than cross-reactivity in some patients.


A new lymphocyte-activating determinant locus expressed on T cells, and mapping in I-C subregion. A new lymphocyte-activating determinant (Lad) locus expressed on T cells was identified, mapping in the I-C subregions of H-2k and H-2d haplotypes. The mixed lymphocyte reaction stimulation could be inhibited by anti-Ia sera made in strains incompatible for this chromosomal segment. Experiments with purified lymphocyte cell populations suggested that this Lad locus was expressed on T cells. Further, only purified T cells were able to remove the inhibiting activity from the anti-Ia sera. I-C subregion gene(s) seem to code for products selectively expressed on a subpopulation of T cells.


Effect of colchicine on the antibody response. I. Enhancement of antibody formation in mice. Colchicine (CC) enhances the antibody response in mice to protein antigens, like diphtheria toxoid and human gamma globulin, as well as to the 2,4,6-trinitrophenyl hapten. Maximal enhancement was observed when CC was administered to animals on the same day as the injection of antigen. The optimal dose of CC was in the range of 1.0-1.5 mg/kg body weight. The enhanced antibody formation was evident from elevated circulating antibody titers and from an increased number of antibody plaque-forming cells (PFC) of the spleen. The circulating antibody titer of CC-treated animals was higher than that of control animals by a factor of about 3-7 in the primary response, and by a factor of at least 15 in the secondary response. In terms of the number of antibody forming cells, CC enhanced the primary PFC response by approximately equal to 100%, and the secondary PFC response by as high as fivefold. The enhancing effect of CC seemed to be related to its mitosis-blocking capacity since (a) vinblastine another antimitotic drug, was found to be as effective as CC and (b)  lumicolchicine, the non-anti-mitotic structural isomer of CC, was ineffective in potentiating antibody responses. The critical timing in the administration of CC on the same day as antigen suggests that most likely, the mitotic poison was acting on antigen-stimulated early dividing suppressor cells.


Localization of T25 glycoprotein in wild-type and Thy 1- mutant cells by immunofluorescence and immunoelectron microscopy. The wild-type BW5147 (Thy 1+) cell line and its Thy 1- mutant derivative BW5147 (Thy 1-a) were examined by immunofluorescence and immunoelectron microscopy for the presence of T25, the glycoprotein which bears the Thy 1 alloantigen. The wild-type cell had T25 predominantly localized on the cell surface. In the mutant cell line, T25 accumulated intracellularly and was present in a clustered distribution throughout the cytoplasm. T25 was not present on the surface of the mutant cell line in significant amount.


Genetic control of the immune response to staphylococcal nuclease. VIII. Mapping of genes for antibodies to different antigenic regions of nuclease. Antibodies to staphylococcal nuclease have been fractionated into two populations on the basis of their ability to bind to the cyanogen bromide cleavage product of nuclease comprising the C-terminal portion of the molecule from the 99th to the 149th amino acid. The two populations of antibodies, anti-nuclease (1-99)n and anti-nuclease (99-149)N, have been prepared from a variety of strains, and analyzed using anti-idiotypic antisera raised against whole anti-nuclease antibodies from strains A/J, SJL, BALB/c, and B10.A(2R). Anti-nuclease (1-99)n, antibodies had the same pattern of reactivity with the anti-idiotypic antisera as did unfractionated antibodies, whereas a different pattern was found for anti-nuclease (99-149)n preparations. On the basis of these studies, five anti-nuclease idiotypes, designated NASE markers, have been identified and defined on the basis of their antigenic specificity and strain distribution. With these additional markers, it has been possible to provide more detailed maps of variable (V) region genes in the strains BALB/c, CB.20, and the recombinant BAB.14. A recombinational event between V region genes during the development of the BAB.14 strain is suggested by the positioning of these NASE markers.


Post-exposure local treatment of mice infected with rabies with two axonal flow inhibitors, colchicine and vinblastine. Post-exposure protection of rabies-infected mice was observed by proximal application of axonal flow inhibitors, particularly vinblastine, to the local nerve(s). These observations indicate that rabies virus is transported by the axonal flow of the peripheral nerves to the central nervous system. Both a fixed virus (CVS) and a street (sylvatic) virus were used. This model in mice could be used to develop an additional post-exposure local treatment of rabies infection in man, by infiltrating local nerves or ganglions with axonal flow inhibitors, with the advantage that it would not interfere with subsequent vaccination as is the case with the administration of hyperimmune serum or immunoglobulin.


Clinicopathological study of Refsum's disease with particular reference to fatal complications. The clinical and pathological findings in two brothers with biochemically diagnosed Refsum's disease are given. The pathology, in general, was that already described in this condition. An unusual complication in one case was the development of renal failure. Death was caused in the other by heart failure.


Effect of ischemia on axonal transport of choline acetyltransferase and acetylcholinesterase and on ultrastructural changes of isolated segments of rabbit nerves in situ. Axonal transport of acetylcholinesterase (AChE) and choline acetyltransferase (ChAc) and ultrastructural degenerative changes were compared in isolated nerve segments of rabbit peroneal nerves kept in vivo for 22 h, either with preserved blood supply (control segments) or under conditions of ischemia (ischemic segments). Ischemia abolished the proximo-distal and disto-proximal axonal transport of AChE and the proximo-distal transport of ChAc which, in control segments, were revealed by accumulations of the enzymes at corresponding ends of the segments. Total activities of AChE and ChAc recovered in isolated segments with intact blood supply corresponded to the activities in normal nerves; in ischemic segments, 50% of ChAc activity was lost in 22 h, whereas all AChE activity was preserved. Ultrastructural changes were found in few fibres in control segments and in many fibres in ischemic segments 22 h after nerve interruption. The early changes in control segments correspond to those described in the literature for peripheral stump of severed nerves. The microtubules, neurofilaments and mitochondria were not affected. In ischemic segments, various stages of axoplasmic disintegration occurred in the myelinated and unmyelinated axons:flocculation and clumping of axoplasmic material, decomposition of neurofilaments and microtubules, swelling, formation of amorphous densities and breakdown of mitochondrial cristae. Swelling, amorphous densities, clumping of nuclear chromatin and necrotic mitochondrial changes appeared also in Schwann cells. It is concluded that ischemia blocks axonal transport and brings about, within 22 h, ultrastructural changes both in nerve fibres and in Schwann cells. Cytoplasmic ChAc is affected earlier by necrotic degeneration of the axons than membrane-bound AChE.


The normal cerebrospinal fluid proteins identified by means of thin-layer isoelectric focusing and crossed immunoelectrofocusing. The cerebrospinal fluid and serum proteins from 32 healthy individuals have been examined by isoelectric focusing in a pH-gradient from 3.5--11.0. Seventeen normal proteins have been identified by means of crossed immunoelectrofocusing. The pI-values of the main fractions of these proteins have been determined. On crossed immunoelectrofocusing microheterogeneity was observed for all of the proteins except haemopexin, gamma-trace protein of CSF and prealbumin of serum. Differences between serum and CSF were observed for 6 of the proteins. Prealbumin of CSF had a lower pI-value than in serum and showed partial immunological identity with prealbumin of serum. Transferrin of CSF included several components with lower sialic acid contents than in serum. The conversion of C'3-complement from beta-1-C to beta 1-A was slower in CSF than in serum. Qualitative differences between CSF and serum were also noticed for alpha2-macroglobulin and haptoglobin. IgM was not detected in CSF. In CSF the gamma-trace protein was found on isoelectric focusing in one third of the subjects. Two subjects had 2 close tau-fractions and another two had one unidentified band in the anodal IgG-area. The gamma-trace protein did not show any proteolytic activity, nor any resemblance in charge to 4 basic enzymes or with the histones tested.


Malignant glioma and sensorimotor neuropathy. A case of sensorimotor neuropathy in a male with malignant glioma is reported. The symptoms of peripheral motor and sensory disturbances preceeded those of the intracranial tumor. The history, clinical findings, electrophysiological and histopathological results are presented, as well as immunological data. A possible causal relationship between glioma and peripheral neuropathy is discussed.


Changes in carotid flow velocity induced by lowering cerebrospinal fluid pressure in normal pressure hydrocephalus. This is a report of changes in blood flow velocity in the carotid system induced by lumbar puncture in five patients who had clinical and neuroradiological signs of normal pressure hydrocephalus. After lowering cerebrospinal fluid pressure an increase of carotid flow velocity was found on Doppler hematotachography. These changes of carotid blood flow velocity could not be demonstrated in a control group of four patients with normal cerebrospinal fluid dynamics. This method is easy to perform, does not overtax the patient and seems to be indicated for the diagnosis of NPH. It is presented as a new, simple, reproducible aid to the diagnosis of NPH by Doppler hematotachography.


Multiple sclerosis and malignant neoplasms in the central nervous system: a clinical anatomical report of three cases. There is some evidence that the incidence of malignant neoplasms in the central nervous system of patients with multiple sclerosis (MS) is more infrequent than amoung the general population. The examination of 3 MS cases with different type of neoplasms in the central nervous system revealed mainly inactive demyelinated plaques. These plaques and the neoplastic lesions were dispersed diffusely and coincidentally all over the brain and spinal cord. The benign course of MS in all three cases is discussed in relation to the malignant diseases.


Myoclonic attacks induced by L-dopa and bromocryptin in Parkinson patients: a sleep EEG study. Six patients with Parkinson's disease developed nocturnal myoclonic attacks after prolongued treatment with L-Dopa which were electroencephalographically recorded. These symptoms persisted after treatment with 2 bromo-alpha-ergocryptin (Bromocryptin), a dopamine receptor agonist, which was substituted for L-Dopa. Bromocryptin is known to have no pre- or postsynaptic effect on serotonin metabolism. It is proposed that these myoclonic phenomena are the expression of the hypersensitivity of denervated catecholamine receptors in the brainstem to the stimulation of L-Dopa and Bromocryptin. This thesis differs with previous suggestions that serotonin plays a major role in the genesis of myoclonic seizures in Parkinsonian patients treated with L-Dopa.


Intrafamilial variability of X-linked progressive muscular dystrophy. Mild and acute form of X-linked muscular dystrophy in the same family. Four of five afflicted boys in the family K. suffer from the Becker type of dystrophy and one from a more severe type. All affected boys and their mothers, who are three sisters, have undergone clinical, electromyographic, electrocardiographic and biochemical examination; muscle biopsy was performed in some boys. This family is a rare example of the intrafamilial variability of X-linked progressive muscular dystrophy. The possible explanation of the variability observed is discussed.