[Remarks on fluorescence microscopy of cerebrospinal fluid cells (author's transl)]. Fluorescencemicroscopic investigations of the liquor cells were carried out by acriclinorange, trypaflavea, the fluorescence component of anilinblue and morin in connection with thiacinred. After that it will be reported about microscopphotometrical measure ments and examinations of immunfluorescence and emphasized the importance of these methods for cytochemistry of the cerebrospinal fluid.


Immunoperoxidase localization of treponema pallidum: its use in formaldehyde-fixed and paraffin-embedded tissue sections. The spirochete, Treponema pallidum, the causative agent of syphilis, has been successfully localized in formaldehydefixed and paraffin-embedded tissue sections using rabbit anti-T pallidum antiserum with two immunoperoxidase techniques. These techniques, the indirect peroxidase-labeled antibody method and the peroxidase-antiperoxidase (PAP) method are compared for sensitivity and degree of nonspecific staining. Both offer substantial advantages over conventional silver-impregnation techniques, but the indirect peroxidase-labeled antibody method seems better, based on the intensity of staining and the simplicity of procedure.


Patterns of hepatitis B surface antigen. Localization in cells of hepatocellular carcinoma. Hepatitis B surface antigen (HBsAg) was identified with immunofluorescence, immunoperoxidase, and aldehyde fuchsin stains within tumor cells in three cases of hepatocellular carcinoma (HCC) from a series of liver biopsies from 172 consecutive cases of HCC. Two patterns of distribution and staining of HBsAg in cells of HCC were observed. In two of the three biopsy specimens, HBsAg was confined to solitary or small groups of tumor cells where a heavily stained inclusion occupied the entire cytoplasm displacing the nucleus. These inclusions corresponded to ground-glass cytoplasm with hematoxylin-eosin. The pattern is different in the other specimen where all the HCC cells in one area of the tumor showed a diffuse peripheral or perinuclear staining of the cytoplasm. In hematoxylin-eosin sections, these tumor cells showed partial transformation of the cytoplasm into the ground-glass appearance.


Preoperative chemotherapy for initially unresectable hepatoblastoma in children. Survival in two cases. In two children with hepatoblastoma, response to chemotherapy permitted subsequent surgical resection. Initially, in both patients, the tumor was thought to be unresectable because of size and/or extent. Chemotherapy with vincristine sulfate and cyclophosphamide resulted in substantial reduction in tumor size, and laparotomy for excision was performed after nine months and four months of therapy, respectively. In case 1, the tumor was localized in the right hepatic lobe and the right hepatic lobectomy was performed. In case 2, the tumor was located in both right and left lobes, but was encapsulated and it was possible to enucleate the tumor with complete gross excision. Both children are living and well without evidence of recurrence of tumor 11 and 23 months after operation.


[Electron microscopic study of archival material fixed in formalin]. A method for treatment of formalin-fixed and paraffin-embedded material for electron microscope examinations is described. Archive material may be examined in the electron microscope for the elucidation of some problems concerning conditions of intercellular structures, comparative size and localization in the cells of secrete granules, myofibrillar apparatus and various inclusions.


Subarachnoid space of the CNS, nasal mucosa, and lymphatic system. We have briefly reviewed the literature pertaining to the movement of tracer molecules and infectious organisms within the olfactory nerve. There is a body of evidence indicating that tracers placed in the CSF will quickly move via the olfactory nerve to the nasal mucosa and then to the cervical lymph nodes. Organic and inorganic tracer materials and organisms as diverse as viruses, a bacillus, and an amoeba, when placed in the nasal cavity, have been shown to move from the nasal mucosa via the olfactory nerve to the olfactory bulb and the CSF. We think that a portion of the data on tracer movement is due to incorporation of tracer materials and organisms into the axoplasm of the olfactory neurons with subsequent anterograde or retrograde axoplasmic transport. However, some of the movement of tracers may occur within the olfactory perineural space. This space may be continuous with a subarachnoid extension that surrounds the olfactory nerve as it penetrates the cribriform plate. To our knowledge, no one has yet followed the perineural space to determine if it is continuous from olfactory receptor to olfactory bulb. The consideration of this space and its role is the main reason for this review.


Ovarian teratoma with peritoneal gliomatosis. The case report describes the clinical and pathological features of a young girl with ovarian teratoma which showed a predominance of neural tissue. The peritoneal involvement by an admixture of mature and immature glial tissue resulted in the death of the patient despite surgery and cytotoxic chemotherapy. There was an association between this tumour and the presence of carcinoembryonic antigen and alphafetoprotein in the serum and ascitic fluid.


Deposition of beta 1H globulin in kidneys of patients with immune renal disease. Renal biopsies from patients with a variety of immune type renal diseases were examined by immunofluorescence for the presence of C3 and its control protein, beta 1H globulin. Deposits of beta 1H were found in every instance (21 of 21 biopsies) in which C3 deposits were observed, irrespective of the underlying disease resulting in the C3 deposits, including lupus nephritis, acute poststreptococcal glomerulonephritis, idiopathic membranous glomerulonephritis, and Goodpasture's syndrome. In no instance was beta 1H found independently of C3. As previously shown in in vitro systems, beta 1H also binds to C3, presumably C3b, during activation of the complement system in immunologically induced renal disease.


Comparison of chlorambucil, azathioprine or cyclophosphamide combined with corticosteroids in the treatment of lupus nephritis. 163 patients with diffuse lupus glomerulonephritis, proven by renal biopsy, were divided into four therapeutic trial groups: 67 were put on corticosteroids alone, 11 on corticosteroids and azathioprine, 32 on corticosteroids and cyclophosphamide, and 53 on corticosteroids and chlorambucil and were followed up for several years. The addition of azathioprine to corticosteroids did not increase the survival rate, improve the renal function or alter the grim prognosis of the patients. Cyclophosphamide appeared to influence favourably the pathological lesion and the renal function when added to corticosteroids, and the disease progressed at a slower rate. The fatal side effects nearly balanced the therapeutic value of cyclophosphamide. Patients on corticosteroids and chlorambucil had an excellent course. This therapeutic regimen resulted in resolution or regression of the renal pathology, marked improvement of the renal function and marked improvement of the survival rate. The authors believe that this therapeutic regimen holds the best chance of becoming the standard treatment for lupus nephritis, particularly since the side effects of chlorambucil were minimal.


Identification of a pure splenic form of chronic lymphocytic leukaemia. We have recently proposed a new staging system for chronic lymphocytic leukaemia (CLL) in which patients with isolated splenomegaly are classified into a distinct stage (stage II). Twenty-three such patients (from two institutions) have been studied without recorded death in a follow-up of 18 months to 30 years. This favourable prognosis justifies separation of these 'pure splenic forms' (SCLL) which must be distinguished from what Galton has termed prolymphocytic leukaemia (PL). This distinction can be made on the basis of three criteria: (i) Clinically, SCLL has a slow uneventful course and neither anaemia and/or thrombocytopenia: (ii) cytologically PL can be distinguished from other forms of CLL though atypical forms of CLL may be confused with the former; and (iii) the study of surface membrane immunoglobulins (SmIg) showed that while lymphocytes from most patients with both PL and SCLL bore uniform SmIg, suggesting a monoclonal B-cell proliferation, there was a major quantitative difference in that whereas PL lymphocytes had a number of antigenic sites close to that of normal lymphocytes (mean: 82 000 sites per cell), SCLL lymphocytes had a drastically reduced number of sites. It is our opinion that this is an important criterion for the differential diagnosis between PL and SCLL.


Stem cell migration induced by erythropoietin or haemolytic anaemia: the effects of actinomycin and endotoxin contamination of erythropoietin preparations. The injection of erythropoietin or the induction of anaemia with phenylhydrazine leads to changes in murine pluripotent and granulocyte-macrophage stem cells indicating migration from marrow to spleen. In order to evaluate the interrelationship between erythroid differentiation and stem cell migration we have selectively suppressed erythroid differentiation with actinomycin D. Anaemia or EP injection resulted in stem cell changes consistent with migration; actinomycin blocked these changes in anaemic but not EP injected mice while blocking erythropoiesis in both groups. The erythropoietin contained from 0.01 to 1000 microgram/ml of endotoxin as defined by the limulus test; it decreased marrow erythropoiesis and stimulated marrow granulopoiesis. Adsorption of the erythropoietin preparation with limulus lysate removed endotoxin without decreasing erythropoietin activity. Adsorbed erythropoietin stimulated erythropoiesis and not granulopoiesis, and stem cell changes induced by its administration were largely blocked by actinomycin, suggesting that endotoxin in the non-adsorbed erythropoietin caused the actinomycin resistant stem cell changes. The observation that actinomycin blocks both erythroid differentiation and stem cell migration suggests that these two physiologic events are closely linked. The effects of injected erythropoietin on murine haemopoietic stem cells may, to a significant extent, be secondary to the presence of endotoxin in the erythropoietin preparations.


Evidence for the platelet specificity of beta-thromboglobulin and studies on its plasma concentration in healthy individuals. The concentration of normal human platelet beta-thromboglobulin (beta-TG) was measured in various washed organ samples by a radioimmunoassay. As only trace amounts were detected, beta-TG appears to be a platelet specific protein. Assay of beta-thromboglobulin in plasma samples from 180 normal individuals gave a range of 10--65 mg/ml. In the 10 subjects studied, plasma beta-TG concentration was related to platelet lifespan but not to turnover. The plasma beta-TG concentration rose with increasing age but did not correlate with the whole blood platelet count or the percentage of megathrombocytes. These results provide further substantial evidence that measurement of plasma beta-TG concentration is useful for assessing the participation of platelets in various disease processes.


Maternal serum alpha-fetoprotein and diabetes mellitus. Maternal serum alpha-fetoprotein (AFP) levels were measured from 12 to 24 weeks gestation in 27 singleton pregnancies in women with insulin-dependent diabetes mellitus and 90 controls without diabetes who were matched for gestational age. The geometric mean AFP level among the diabetic pregnancies was 60 per cent of that in the controls, a difference which was statistically significant (P less than 0.01). The difference in AFP level between diabetic patients and controls was most marked in sera taken before 21 weeks gestation.


Maternal serum alpha-fetoprotein screening in a provincial Health District. Maternal serum alpha-fetoprotein (AFP) estimation as a routine screening test for neural tube defect (NTD) was introduced into the West Berkshire Health District, where 17 per cent of all deliveries and most of the antenatal care is undertaken by the general practitioners. In the first year, 4458 patients were screened and 43 of those (0.96 per cent) had raised serum AFP levels. Amniocentesis was performed on 31 patients (0.69 per cent). Ten fetuses with severe NTDs and one with exomphalos were detected and the pregnancies terminated. In six patients, raised serum AFP levels were due to fetal death. No normal pregnancy was terminated. Acceptability by patients was high. Provided that a good diagnostic ultrasound facility is available locally, maternal serum AFP estimation seemed to be a valuable screening test.


Antenatal screening in Oxford for fetal neural tube defects. Between May 1975 and the end of 1977, 6443 antenatal patients were screened mainly between 16 and 22 weeks of pregnancy for neural tube defects (NTDs) at the John Radcliffe Hospital, Oxford, by maternal serum alpha-fetoprotein (AFP) measurement; a take-up of 72 per cent. Seventeen out of 18 (94 per cent) patients with open NTD pregnancies (9 out of 9 with anencephaly and 8 out of 9 with open spina bifida) had positive screening tests, and all except one were offered and accepted a termination of pregnancy. Two hundred and forty-five (3.8 per cent) patients with unaffected pregnancies also had positive screening tests, although only 1.4 per cent had an amniocentesis. Following ultrasonography, about 50 per cent of patients with unaffected pregnancies with positive screening tests were not offered an amniocentesis because they had a multiple pregnancy or their gestational age had been underestimated. The odds of having a fetus with an NTD among the women who had an amniocentesis was about 1 to 6 (1 to 11 for open spina bifida alone). Two apparently normal pregnancies were terminated. A survey of the acceptability of the screening programme among a consecutive sample of 73 patients who knew that they had a positive screening test revealed that all except one had no objection to screening in general, and 68 (93 per cent) wanted to be tested again in a future pregnancy. The approximate direct cost of the programme was 2 pounds to 3 pounds per patient screened, or about 1000 pounds per NTD detected (about 2200 pounds per open spina bifida detected).


Clinical dilemmas arising from the antenatal diagnosis of neural tube defects. Eight case histories are presented which demonstrate clinical problems associated with the antenatal screening for and diagnosis of neural tube defects. It is suggested how some of these problems might be avoided in future.


Evidence for two forms of RNA-dependent DNA polymerase in Visna virus. The visna viral RNA-dependent DNA polymerase has been resolved into two forms by affinity chromatography. Glycerine gradient centrifugation of the two forms showed that one form sedimented at 6.9 S corresponding to an apparent molecular weight of 135 000 and the other at 6.3 S corresponding to 118 000. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of the two forms indicated that the 6.9 S enzyme is composed of 2 molecules of 68 000 mol. wt. chain and the 6.3 S is a single chain enzyme. The latter form has been identified as a glycoprotein. The 6.9 S form can be completely inactivated in 20 min at 45 degrees C, prefers poly(rC) over poly(rA) as template and has high efficiency in utilizing visna 70 S RNA as template. The 6.3 S form is stable at 45 degrees C, active with 70 S viral RNA as template, prefers poly(rA) over poly(rC), and requires higher concentration of Mn2+ (0.4 mM) for maximum activity than the 6.9 S form does (0.1 mM) with synthetic homopolymers as templates. However, both 6.9 S and 6.3 S forms prefer Mg2+ over Mn2+ regardless of the nature of the templates.


The effect of alkaline earth cations on the release of histamine from rat peritoneal mast cells treated with compound 48/80 and peptide 401. 1 Extracellular calcium ions have a dual effect on the release of histamine from rat peritoneal mast cells treated with compound 48/80 and peptide 401. The release is either potentiated or inhibited according to the relative concentrations of ion and inducer.2 Strontium similarly potentiates the release produced by optimal concentrations of inducer but higher concentrations are required than in the case of calcium. Strontium is markedly less inhibitory than calcium.3 Mast cells may be depleted of intracellular calcium by incubation for short periods with the chelating agent, ethylenediamine tetraacetic acid (EDTA). They thereby become unresponsive to compound 48/80 and peptide 401 unless calcium is reintroduced into the incubation medium. Strontium and barium, but not magnesium, will substitute for calcium in this system. Barium additionally produces a marked release of histamine even in the absence of inducer. Pretreatment with the ionophore A23187 similarly inhibits the subsequent response to peptide 401 in divalent cation-free medium. This inhibition is reversed on the reintroduction of calcium.4 Compound 48/80 and peptide 401 release histamine from mast cells incubated in isotonic sucrose in the complete absence of added metal ions. However, the corrected release under these conditions is potentiated by both mono and divalent cations.5 On the basis of these results, the possible mechanism of action of the basic releasing agents and their usefulness as models for studying histamine secretion is discussed.


Dorsal column nuclei and ascending spinal afferents in macaques. Cell populations and thalamic projections of the dorsal column nuclei in macaques have been investigated in the medullae of normal animals and of animals with injections of horseradish peroxidase in the nucleus ventralis posterolateralis. In the same species, the course, distribution and origin of ascending non-primary pathways to the dorsal column nuclei have been demonstrated with the aid of degeneration methods, 3H-amino acid autoradiography and retrograde axonal transport of horseradish peroxidase. Non-primary afferents to the gracile and cuneate nuclei ascend mainly in the dorsal columns and, to a lesser extent, in the dorsal part of the lateral funiculus. Afferents originating from lumbar segments and ascending in the lateral funiculus terminate mainly in the rostral part of the gracile nucleus while those ascending in the dorsal columns distribute throughout most of the rostrocaudal extent of the same nucleus. Afferents from brachial levels terminate mainly in the cuneate nucleus and in the external cuneate nucleus. Degeneration and autoradiographic material concurrently demonstrate that non-primary afferents to the cuneate nucleus terminate preferentially within certain cytoarchitectonic subdivisions of this nucleus. Ascending spinal afferents to the dorsal column nuclei originate mainly from the ipsilateral dorsal horn, particularly from its medial part at upper cervical levels and from a band of gray, throughout the cord, largely corresponding to lamina IV and adjacent laminae. Large neurons along the lateral border of the ventral horn at lumbar levels may also contribute non-primary afferents to the ipsilateral dorsal column nuclei. These anatomical results provide some cues to a revised view of the organization of the dorsal column nuclei in monkeys and, taken together with recent electrophysiological and clinical data, contribute to a re-evaluation of some functional aspects of the dorsal column-medial lemniscal system of primates.


A study of the morphology of the large reticulospinal neurons of the lamprey ammocoete by intracellular injection of Procion yellow. 47 reticulospinal neurons of the lamprey brain-stem were stained by intracellular injection of Procion yellow, so that their morphology and dendro-architecture could be determined. The vagal, bulbar, Mauthner, isthmic and mesencephalic groups could be distinguished by the shape and size of their somata and the distribution of their dendrites. However, they shared the following features: the somata of all these cells were relatively large (up to 80 micrometer largest diameter); the dendrites were extensively distributed in the brain-stem in an open-field arrangement; the initial segment expanded abruptly at the soma junction but tapered gradually to form a narrow axon, which joined the medial longitudinal fasciculus in the brain-stem.


Nerve growth factor action on 2-deoxy-D-glucose transport in dorsal root ganglionic dissociates from chick embryo. Dorsal root ganglionic cells, when incubated in vitro in the absence of nerve growth factor (NGF), undergo a general metabolic degeneration which is preceded by loss of certain permeation properties. To determine in which ways an absence of NGF can also affect the capacity of these cells to take up an important energy source, namely glucose, experiments were carried out in which cells were incubated with or without NGF for varying times, and then presented with the factor and tested for the ability to take up 3H-labeled 2-deoxy-D-glucose. As with exogenous uridine, hexose transport in DRG cells was reduced by NGF deprivation and restored by delayed NGF administration (up to 6 h). Both the initial rate and equilibrium level were affected in an NGF dose-dependent fashion. Calculation of apparent Kt and V max in NGF-deprived and NGF-supported cells showed about two-fold differences between NGF-controlled and NGF-independent hexose transports, suggesting corresponding differences between NGF-dependent and other ganglionic cells. Restoration of hexose transport by delayed NGF administration took place within minutes of presentation of the factor. The delay before onset of restoration and the speed with which restoration was achieved have been found also to be dependent on the NGF concentration, suggesting that they reflect equilibration kinetics between NGF and its binding sites rather then the development of the response within cells. Thus, NGF can rapidly modulate permeation properties which regulate the availability of major energy substrates for the cell. This effect of NGF is discussed in the content of current views on the mode of action of the factor.


Axonal transport of taurine along neonatal and young adult rat optic axons. Studies in this laboratory have indicated that taurine is axonally transported along goldfish optic nerves. In the present experiments the axonal transport of taurine was examined in neonatal and young adult rat optic axons. [35S]taurine was injected into the vitreous humor of right eyes of developing (1--15-day-old) or young adult (40-day-old) rats. At various times after injection ranging from 3 h to 7 days, right retinae and left and right geniculates were removed and assayed for radioactivity, left minus right lateral geniculate (L-RLG) radioactivity being used as an index of axonally transported [35S]taurine. Results indicated that taurine was rapidly transported along both neonatal and young optic axons, in contrast to other amino acids (i.e., leucine and proline) which are not axonally transported in this system. Significant developmental variations were seen in both L-RLG and right retinal [35S]taurine activity 24 h after injection. The amounts of L-RLG [35S]taurine corrected for retinal ganglion cell uptake in animals injected at 1,4,7 and 11 days after birth (prior to and during the major period of synaptogenesis in the geniculates) were 4.5, 3.1, 2.3 and 2.6 times higher, respectively, than those in the young adults. In contrast, the amount of corrected L-RLG [35S]taurine in animals injected at 15 days after birth (after synaptogenesis) were not significantly different from that in the young adult.


Radiation therapy for localized prostate cancer. Since 1965, 401 patients with prostate cancer have received intensive local pelvic radiation therapy at the Virginia Mason Medical Center. Two hundred seventy-seven of these patients were treated from 1965 through 1975, comprising the study group. Two hundred twenty-one of this series were in the Stage C category. The 36 Stage B cancers were either medically nonoperable, or advanced extent, or had high-grade histopathology. Ten patients each were in diffuse Stage A or Stage D groups, the latter receiving local palliative intensive treatment to the prostate area. The mean age of the patients was 67.6 years. The five year survival of the Stage C group was 57.7%.  There was no apparent influence on the survival of irradiated Stage C patients who received estrogen therapy. Current treatment techniques employ 10 megavolt photon beam with whole pelvic nodal fields and bilateral arc rotational boost fields. The incidence of reactions and complications is presented.


Unusual sensitivity of ataxia telangiectasia cells to bleomycin. Peripheral blood lymphocytes from four patients with ataxia telangiectasia (AT), an inherited disorder showing, among other features, radiosensitivity and a high frequency of cancers, were shown to be cytogenetically more sensitive to bleomycin than were lymphocytes from both normal individuals and a single patient with xeroderma pigmentosum. With cell survival techniques, a biphasic dose-response curve was seen for both normal and AT fibroblasts, although the AT cells showed a much lower survival. The increased sensitivity to bleomycin in AT cells might be expected since it is a radiomimetic drug, but more importantly the known action of bleomycin in producing DNA strand scission suggests that AT cells might be defective in rejoining a proportion of DNA strand breaks.


Preparation of permanent slides of intact soft-agar colony cultures of hematopoietic and tumor stem cells. A simple technique is described for fixing colony-containing layers of soft agar and drying them onto microscopic slides. The method is extrapolated from techniques used in immunology for permanent preservation of immunodiffusion or immunoelectrophoresis plates. Slides prepared in this fashion are eminently suitable for subsequent analysis with a variety of techniques including conventional Papanicolaou or other staining methods as well as histochemistry, immunofluorescence, and autoradiography. In addition to research applications, the technique may have diagnostic applications and should greatly enhance both qualitative and quantitative analysis of the biology of hematopoietic and tumor colony formation.


Silver staining of nucleolar granules in tumor cells. With the aid of a simple silver-staining procedure, large numbers and unusual arrays of nucleolar argyrophilic granules were found in Novikoff hepatoma, KB, and HeLa cells. Some of these arrays consisted of linearly arranged discrete granules, and others were in two to three rows each containing three to five granules. Corresponding formations were not found in either the normal or regenerating liver nucleoli which contained an argyrophilic network in which the dark granules were apparently associated with the less dark argyrophilic fibrils of a reticulum. The nucleolar argyrophilic granules were readily identifiable in the separated daughter nuclei of the tumor cells in telophase, suggesting that the increased nucleolar activity of the G1 phase begins in these cells even before cell division has been completed.


Antiproliferative effects of 9-beta-D-arabinofuranosyladenine 5'-monophosphate and related compounds in combination with adenosine deaminase inhibitors against mouse leukemia L1210/C2 cells in culture. The antiproliferative activity of 9-beta-D-arabinofuranosyladenine 5'-monophosphate against a cultured line of mouse leukemia cells (L1210/C2) was enhanced by addition of either 2'-deoxycoformycin or erythro-9-(2-hydroxy-3-nonyl)adenine. The activity of 9-beta-D-arabinofuranosyladenine 5'-monophosphate, alone or in combination with either of the two inhibitors of adenosine deaminase, was comparable to that of 9-beta-D-arabinofuranosyladenine (ara-A), apparently reflecting the rapid conversion of 9-beta-D-arabinofuranosyladenine 5'-monophosphate to ara-A by L1210/C2 cells. Several ara-A analogs were assayed for antiproliferative activity against L1210/C2 cells; of these, only 9-beta-D-arabinofuranosyladenine 5'-O-methylphosphate and 2'-deoxy-2'-amino-9-beta-D-arabinofuraosyladenine were active. Addition of 2'-deoxycoformycin to cell culture fluids enhanced the activity of 9-beta-D-arabinofuranosyladenine 5'-O-methylphosphate suggesting conversion to an adenosine deaminase-sensitive intermediate, presumably ara-A.


Changes in peroxisomes in preneoplastic liver and hepatoma of mice induced by alpha-benzene hexachloride. Peroxisomes in hepatomas and hyperplastic preneoplastic liver lesions induced in mice by 500 ppm alpha-benzene hexachloride were examined histochemically and electron microscopically. Although most of the hepatomas were well-differentiated tumors and contained a considerable number of peroxisomes, the tumor cells did not respond to ethyl-alpha-p-chlorophenoxyisobutyrate with proliferation of peroxisomes. At the 16th week of carcinogen feeding, hyperplastic nodules appeared and advanced to further stages. A majority of the nodules showed a considerable number of peroxisomes and the inductive proliferation of peroxisomes. Within the nodules, foci of proliferation of the cells that showed no inducibility of proliferation of peroxisomes appeared. These cells proliferated further, replacing the most part of the nodules, and with this process hepatomas appeared to have been formed. No abnormal matrical inclusions of peroxisomes were formed in the cells of hyperplastic nodules by ethyl-alpha-p-chlorophenoxyisobutyrate unlike in the case of rats.


Facilitated light microscopic cytochemical diagnosis of acute myelogenous leukemia. Hydroperoxidase-positive Phi bodies and rods are much more prominent and prevalent than rods visualized with a Romanovsky-type stain (Auer rods) in immature leukocytes of patients with active acute myelogenous leukemia (AML). They are readily observed with the light microscope in peripheral blood or marrow films of AML patients stained to show their peroxidatic activity. In many of these patients, Auer rods, which apparently constitute only a small subpopulation of the hydroperoxidase-positive Phi bodies and rods, were detected with difficulty, if at all. The hydroperoxidase-positive Phi bodies and rods were observed in 92% of 36 patients with active disease. They were never observed in leukocytes of patients with other hematopoietic disorders or of normal individuals. Thus, they facilitated the distinction of AML from acute lymphocytic leukemia and chronic granulocytic leukemia in blast crisis. They were absent in full clinical remission after chemotherapy and were greatly diminished in partial remission. They were present in disease relapse and reappeared in five patients who had been in full remission. These results suggest that these hydroperoxidase-positive enlarged particles are pathognomonic of AML and that monitoring them with the light microscope may aid in guiding its clinical management.


Specific cellular immune responses in patients with malignant gliomas. The leukocyte adherence inhibition assay was used to measure cell-mediated immunity in 26 patients with malignant glial neoplasms and 41 control subjects. A significant inhibition of leukocyte adherence was observed in 21 of 26 (80%) glioma patients in the presence of a 3 M KCl extract of glioma tissue, as compared to that of normal brain extract. Among the control group, no significant difference in the percentage of nonadherent leukocytes was noted in the presence of either antigen. To study the specificity of the reaction, a 3 M KCl extract of meningioma, pituitary tumor, carcinomas of breast, and lung, melanoma, brain, and heart tissues were used as nonspecific antigens. Such studies revealed significantly lower values of nonadherent leukocytes. These data indicate that patients with malignant glial neoplasms manifest a cellular immune response to glioma-associated antigens which can be measured by the tube leukocyte adherence inhibition assay and that leukocyte adherence inhibition assay may render additional useful information in diagnostic and prognostic evaluation of malignant glial neoplasms.


Immunological analysis of A strain mice bearing the A-10 mammary adenocarcinoma. The present investigation was undertaken to determine whether antitumor antibodies are produced by A strain mice during the growth of a transplantable mammary adenocarcinoma (A-10). The antibody response was monitored by a sensitive radioimmunoassay which can detect 1 ng of antibody. No evidence of a humoral antitumor response was observed in animals given i.p. or s.c. injections of A-10 ascites cells. Control experiments showed that a humoral response was detectable 1 week after the inoculation of an allogeneic tumor. Immunoglobulin binds nonspecifically to cells via an Fc portion of the immunoglobulin molecule, and this was seen with a tumor bearer serum pool and with immunoglobulin preparations eluted from A-10 ascites cells. No specific antitumor antibody was found in these sources. The A strain mice could not be immunized to reject a challenge of live A-10 cells with mitomycin C-treated A-10 cells, with neuraminidase-treated A-10 cells, or with A-10 membrane preparations. It was concluded that the A-10 tumor is not immunogenic in its host of origin.


Characterization of the carcinoembryonic antigen activity associated with cyst fluids of mucinous ovarian cystadenocarcinoma. Cyst fluids from mucinous cystadenocarcinoma of the ovary show significant concentrations of carcinoembryonic antigen (CEA) activity. This CEA activity was compared to several colon CEA standards with respect to size, concanavalin A binding, and immunological activity. CEA activity in unfractionated ovarian cyst fluid was indistinguishable from CEA standards by gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The cyst fluid CEA activity showed identity with various CEA standards by double immunodiffusion in agar gels and by radioimmune competition assay. Attempts to purify the cyst fluid CEA by perchloric acid extraction prior to gel filtration chromatography were unsuccessful. The yield of soluble CEA activity following perchloric acid extraction averaged 10%, and in one cyst fluid, qualitative changes were noted in the soluble antigenic activity. Ovarian cyst fluid CEA activity was bound by concanavalin A-Sepharose 4B and specifically eluted with competing monosaccharide. Lectin affinity chromatography and gel filtration chromatography over Sepharose 4B and Sephadex G-200 can be used to purify cyst fluid CEA from whole fluid or lyophilized and reconstituted samples.


Heterogeneity in cell-associated transformation-sensitive proteins antigenically related to fibronectin. Antiserum to purified fibronectin has been used to investigate transformation-associated heterogeneity in surface components antigenically related to fibronectin. Examination of extracts from surface-labeled rat cells by gradient gel electrophoresis revealed in "normal" cells the presence of two species with approximate molecular weights of 250,000 and 230,000, which were decreased in wild-type transformed cells. Reaction with antifibronectin serum revealed the selective precipitation of the two transformation-sensitive surface components. A similar experiment with ts-NT3-KR that expresses a normal phenotype at 37 degrees and a transformed morphology at 33 degrees did not reveal a markedly altered surface labeling pattern at both temperatures. However, reaction with antifibronectin serum did show a weak but detectable recognition of a 230,000-dalton doublet in the cells grown at 37 degrees, and immune precipitation of components in the 100,000- and 60,000-dalton region in cells grown at 33 degrees. Experiments with normal mouse cells revealed a different ratio of two fibronectin-related external proteins with similar molecular weights to those seen in normal rat cells.


Combined radiotherapy and multidrug chemotherapy for advanced head and neck cancer: results of a Radiation Therapy Oncology Group pilot study. Between February 1976 and June 1977, 15 patients with advanced inoperable squamous cell carcinoma of the head and neck were entered in a pilot study sponsored by the Radiation Therapy Oncology Group using combined radiotherapy and multidrug chemotherapy. Chemotherapy consisted of cyclophosphamide, vincristine, and bleomycin (during radiotherapy) and cyclophosphamide, methotrexate, and bleomycin (after radiotherapy). At the time of last followup or of death, the disease had been controlled at the primary site in eight of the 15 patients (53%), in the neck in 12 patients (80%), and in all sites in seven patients (47%). Followup time ranged from 2 to 24+ months with a median of 7 months. Acute toxic effects of the combined treatment, primarily enhanced radiation mucositis and infection, were severe, and three patients had fatal complications. Although the combination chemotherapy may have enhanced the tumor response to radiotherapy in some of these patients, treatment morbidity and complications were prohibitive for the treatment program to progress to a randomized trial.


Absence of anthracycline-induced degradation of nuclear DNA in Ehrlich ascites tumour cells. The in vivo effects of anthracycline antibiotics on the integrity of Ehrlich ascites tumour cell DNA have been studied by sedimentation analysis of nuclear structures containing superhelical DNA in neutral sucrose gradients. These fast-sedimenting protein-DNA complexes may be released by gently lysing cells in solution containing non-ionic detergents and high NaCl concentrations (1.95 M). The supercoiled structure of DNA in these protein-DNA complexes is suggested by the characteristic sedimentation in the presence of intercalating agents. Apparently, no DNA damage could be detected in Ehrlich cells from 7-day-old tumours within 3 h after various doses of daunomycin (0.5--10 mg/kg of body wt.) were administered i.p. to mice. Sedimentation anomalies could not be observed even 15 or 30 h after administration of rtherapeutic doses of daunomycin or adriamycin. In contrast, at 30 min after administration to mice, therapeutic doses of bleomycin (2--8 mg/kg) caused extensive fragmentation of tumour cell DNA, which could be monitored as slowly sedimenting DNA structures (compared with the the control). Similarly, DNA damage could be induced by procarbazine at therapeutic doses. Exposure to bleomycin or procarbazine abolished the characteristic biphasic response to ethidium bromide. The absence of anthracycline-induced degradation of Ehrlich ascites tumour cell DNA is apparently in contrast with the DNA damage observed in L1210 tumour cells. These observations suggest that DNA damage is not a necessary condition for antitumour activity.


An experimental study on the effect of plasma expanders on blood coagulability. The effect of various plasma expanders on blood coagulation was studied using the thromboelastography (TEG). Solutions of 5, 10, 20, and 50% concentration of each expander (6% low molecular weight hydroxyethyl-starch (6% Hespander), 3% dextran-40, 6% high molecular weight hydroxyethyl-starch (6 HES), and 6% dextran-70) or solvent (lactated Ringer solution and normal saline) in blood were prepared and their coagulability was examined by TEG. In the low molecular weight plasma expanders (6% Hespander and 3% dextran-40), in general, the coagulability decreased when the concentration was increased. In the high molecular weight plasma expanders (6 HES and 6% dextran-70), the coagulability increased slightly at low and high concentrations and the coagulability was reduced. As a conclusion, at clinically used concentrations of less than 20% in blood, changes in the TEG of the plasma expanders are minimum and have no clinical significance.


Central serotonergic agents raise the repetitive extrasystole threshold of the vulnerable period of the canine ventricular myocardium. Systemic administration of three central serotonergic agents, melatonin, 5-methoxytryptophol, and 6-chloro-2-(1-piperazinyl)-pyrazine (MK-212), produced significant increases in the threshold of the vulnerable period for repetitive electrical activity in the canine cardiac ventricle. MK-212 was effective despite bilateral vagotomy. The specific serotonin antagonist, metergoline, blocked the effect of MK-212 on the threshold. An increase in central serotonergic activity may inhibit the flow of arrhythmogenic sympathetic nerve traffic from the brain to the heart.


Modulation of the properdin amplification loop in membranoproliferative and other forms of glomerulonephritis. The mean serum concentrations of C3b inactivator (C3bINA) and beta1H globulin were measured in eighty-six sera (thirty-five membranoproliferative, thirteen membranous, thirty-one focal and seven minimal change) taken from seventy-five patients with histologically diagnosed glomerulonephritis. In none of the four groups of sera did the mean concentrations of both these proteins differ significantly from the mean levels in a group of fifty normal sera. However, in the membranoproliferative group, the mean level of C3bINA, but not beta1H, was significantly lower in the nephritic factor positive than in the nephritic factor negative group. When the eighty-six sera were considered together, serum concentrations of C3bINA and beta1H correlated with levels of C3, factor B, properdin and C4. When the membranoproliferative group was considered separately, C3bINA levels correlated with levels of C3, factor B, properdin and C4 and beta1H correlated with C3 and factor B, but not properdin or C4 in the nephritic factor negative group, but no such correlations were observed in the nephritic factor positive group. The significance of these findings is discussed in the light of current thoughts on the mechanisms of modulation of the alternative pathway.


Oligoclonal immunoglobulins in subacute sclerosing panencephalitis and multiple sclerosis: a study of idiotypic determinants. Studies have been made of the idiotypic determinants of subacute sclerosing panencephalitis (SSPE) antibodies using rabbit antisera to serum and spinal fluid fractions. Evidence is presented indicating that serum and cerebrospinal fluid (CSF) anti-measles antibodies, as judged by their idiotypes, differ in their relative concentrations in the two compartments. The results indicate that some of these antibody subpopulations originate within the CNS, while others are made largely or entirely outside. In addition to strong idiotypic specificity, a limited cross-idiotypic specificity relating antibodies from three out of fourteen SSPE patients has been identified. In the course of these studies, measles virus was found to agglutinate red cells coated with antibody fraction to high titres. This system has proved useful in demonstrating the competition between anti-idiotypic antibody and antigen for the combining sites of the measles antibody. Two anti-idiotypic antisera have also been obtained against the spinal fluid IgG of multiple sclerosis (MS) patients. The possible use of these marker reagnets as well as related methodologies in the search for the antigens involved in MS bands is discussed.


Mercury poisoning from mercurochrome therapy of an infected omphalocele. A neonate with an infected omphalocele was treated locally with merbromin (mercurochrome) for five days. Extensive skin peeling with bullous lesions, edema, and fever developed three days after mercurochrome therapy. The infant died on the ninth day. Autopsy revealed evidence of heavy metal poisoning of the kidney, excessive mercury levels in the blood, and in tissues of the brain, kidney, and liver.


The visualization of individual collagen molecules and aggregates in collagen solution: the effect of preparation and sampling techniques. The use of electron microscopic observations to characterize the state of aggregation of collagen molecules in solution has been examined from the point of view that the observed structures must be compatible with the physical chemical properties of the solution. The common procedure of addition of droplets of a collagen solution to a grid followed by negative staining produces aggregates on the grid which could not have represented the aggregation state in solution. Closer representation of fibril diameters can be achieved by freeze cleave-etch methods. The best representation of the state of aggregation in collagen solutions can be achieved by use of a careful drop-washing procedure. This procedure, described in detail, permits correlation between intrinsic viscosity and electron microscopic observations.


Detection of heart rhythm disturbances in patients after myocardial infarction, and their prognostic value. For one year 215 patients were followed up who had been taken into the register of patients with myocardial infarction and had survived three months after the incident. At intervals of 3, 6, 9, and 12 months their electrocardiograms of various durations were recorded. Prolongation of the ECG tracing time eases the assessment of the frequencies and nature of heart rhythm disturbances in patients with ischaemic heart disease. Exercise test helps reveal more complicated types of ventricular extrasystoles better than one-hour recording of resting ECG does. Ventricular ectopic activity is more frequently found in patients with unequivocal ECG changes (p less than 0.025), in patients with enlarged heart volume (p less than 0.05), and in men vs. women (p less than 0.05). Presence of ventricular extrasystoles, especially of complicated ones (polytopic, couplets, early) enhances the risk of death of ischaemic heart disease.


Silver-stained structures in mammalian meiotic prophase. Silver staining of mammalian spermatocytes revealed, in light microscopy, synaptonemal complex and structures within the sex vesicle. It is feasible to follow the chromosome pairing phenomenon from zygotene to pachytene by examining the behavior of synaptonemal complexes. Nucleolus organizer regions take heavy silver stain in pachytene but are no longer detectable in later stages of meiosis.


Pattern of activity of nucleolus organizers during spermatogenesis in mammals as analyzed by silver-staining. Silver-staining in the nuclei and chromosomes of spermatogenesis of four species of mammals (Man, Mus musculus, Rattus norvegicus, and Cavia cobaya) was investigated qualitatively and quantitatively. These species show a very similar pattern of activity of the nucleolus organizer regions (NORs) during the various stages of spermatogenesis. Silver precipitates are detectable in growing spermatogonia and up until the pachytene stage of meiotic prophase. During the meiotic metaphases I and II and during interkinesis silver-stainability disappears completely. A resumpton of silver-stainability occurs in round spermatids indicating a postmeiotic reactivation of NORs. This process does not persist beyond the early elongation phase. The quantitative determination of the silver-covered areas in relation to the total nuclear areas reveals minor differences between the species investigated with regard to the times and extents of maximum activation. The known localizations of the NORs in the karyotypes of the species investigated was confirmed using metaphase-preparations derived from somatic tissues.


Failure of endoscopic transmission of hepatitis B. Emergency endoscopy was performed on two patients subsequently found to be hepatitis B surface antigen carriers. Before their carrier state was determined, nine other patients underwent endoscopy using the same instruments, which had been routinely cleaned between procedures. These patients were all notified within five days of the incident, given standard gamma globulin, and prospectively followed for the development of hepatitis. After one of the endoscopes was gas sterilized, the next three patients undergoing endoscopy were also followed. One of the hepatitis B surface antigen carriers was positive for antibody to e antigen; the other carrier had neither e antigen nor antibody. None of these individuals developed signs or symptoms of hepatitis, abnormal serum glutamic pyruvate transaminase elevations, or serologic evidence of hepatitis B exposure. From these data, and other recorded experiences, it appears that routine cleansing of endoscopy equipment is sufficient in preventing the transmission of hepatitis B.


Electroencephalography in brain edema (127 cases of brain tumor investigated by cranial computerized tomography). Cranial computerized tomography (CCT) in permitting visualization of cerebral edema in live patients, allows for the first time valid studies concerning the role of reactional edema in the generation of EEG abnormalities related to expanding processes. The authors analyze the results of EEG and CCT in 127 patients presenting cerebral tumor(s), 84 of which were accompanied by reactional edema. The study leads to the conclusion (also demonstrated by certain animal work) that edema per se is only rarely responsible (9.5%) for the EEG abnormalities.


Electroencephalographic cranial computerized tomographic correlations in brain abscess. The authors studied 19 cases of brain abscess. The investigation included one or more EEG records from 13 patients and one or more cranial computerized tomograms in all cases. In this work the two techniques were associated to establish the diagnosis of cerebral abscess. The EEG is almost always abnormal and pronounced EEG disturbances are, in most cases, sufficient for indicating a diagnosis of a space-occupying process. On the other hand, EEG patterns suggestive of brain abscess were detected in only 38% of the patients. In brain abscess, CCT is of considerable value since the existence of 'suppurative' images was demonstrated in all cases and of these, 70% were characteristic. The association of these two techniques is thus extremely useful in the diagnosis of cerebral abscess, to follow its evolution and to detect postoperative relapse or to evaluate the efficiency of medical management alone.


Effects of changes in cortical excitability upon the epileptic bursts in generalized penicillin epilepsy of the cat. Previous studies had suggested that the epileptic bursts of feline generalized penicillin epilepsy represent the response of hyperexcitable cortex to thalamocortical volleys normally evoking spindles. If this were the case, it should be possible to convert the epileptic bursts of generalized penicillin epilepsy into spindles by decreasing the excitability of cortical neurons. In cats exhibiting the EEG signs of feline generalized penicillin epilepsy cortical excitability was decreased by hypoxia, by the topical application to the cortex of KCl (inducing spreading depression), barbiturates, GABA, AMP or noradrenaline. During generalized penicillin epilepsy, hypoxia and KCl-induced spreading depression abolished epileptic bursts which were replaced by spindles. When spindles and epileptic complexes occurring in the same animal were compared, a direct correlation between the frequencies of these two rhythms could be demonstrated, that of the epileptic complexes being about half that of the spindle waves. These observations support the hypothesis that the epileptic bursts of feline generalized penicillin epilepsy are induced by thalamocortical volleys normally involved in spindle genesis. Topical cortical applications of barbiturates, GABA, AMP and noradrenaline reduced or inverted the negative spikes of the spike and wave complexes, while augmenting the negative slow waves, or revealing them clearly in instances in which they had been poorly developed. This effect is interpreted as being due to a selective inactivation of the superficial cortical layers. That topical cortical application of barbiturates, GABA, AMP and noradrenaline was capable of transforming into typical spike and wave complex epileptic bursts, which had not previously conformed to this pattern, indicates that the intracortical electrophysiological events of typical and atypical epileptic bursts in feline generalized penicillin epilepsy are fundamentally the same and reflect an alternation between excitatory and inhibitory sequences.


Petit mal and grand mal seizures produced by toluene or benzene intoxication in the cat. Motor incoordination, euphoria and hallucinations are symptoms reported for humans voluntarily intoxicated by industrial solvents. An epileptic-like consciousness impairment has also been noted. The present paper describes a technique used for the experimental study of solvent intoxication in which toluene and benzene can be applied directly into the trachea of freely moving cats with chronically implanted electrodes. This technique permits the control of solvent dose and time of exposure. Results showed a 3 Hz spike-wave activity in the gyrus cinguli recording with both toluene or benzene intoxication. Furthermore, benzene inhalation produced generalized tonic-clonic seizures. These effects were dose-related. However, a sensitization period was essential for the development of such alterations, and effects showed a tendency to shortening through chronic exposures. These alterations were correlated with behavioral disturbances such as nodding, twitching and apparent hallucinations. Results are discussed regarding the sensitization period, the optimal peak of effects, and the period of tolerance development relevant to an earlier found amygdalar activation that could be correlated with other methods inducing experimental seizures, such as repetitive stimulation of the brain (kindling).


Fast somato-parietal rhythms during combined focal attention and immobility in baboon and squirrel monkey. In the monkey (baboon or squirrel monkey) displaying a high level of vigilance, a particular type of spontaneous ECoG rhythm develops when the subject is motionless and fixes its attention on a visual target in its surroundings. This activity, whose frequency averages around 18 c/sec, is distributed over two restricted cortical foci, one over the SI hand area (Brodmann's fields 1 and 2) and the other over the posterior parietal cortex (area 5). These rhythms are blocked by the least body movement (but not by eye movements). In view of their location and reactivity, the rolandic portion of these rhythms belongs to the 'mu' type as described in man. The suggestion is made that this synchronized activity favours the maintenance of immobility as well as the fixation of attention on a non-somaesthetic target by temporarily blocking both somaesthetic (in area SI) and somatopraxic integration (in area 5).


Observations on the M-wave and the CNV in the squirrel monkey. A typical CNV paradigm, with food as reward, evokes in the squirrel monkey's post-arcuate and post-central cortices both M-waves in response to the cues and what appears to be a CNV in the interstimulus interval. Both wave forms appear to be generated locally in cortex and to be more closely related temporally to the cues than to the animal's behavioral response. The CNV, like the M-wave, appears to reflect the animal's level of interest in obtaining the reward. It is smaller when the animal misses the cues than when he responds correctly. It decreases as food pellets are consumed. It varies with the level of interest of the animal in a particular type of food pellet, as judged from the animal's behavior. At some recording sites in some animals it appears to reflect both interest in the reward and interest in manipulating the environment to obtain the reward. It can be maintained for at least 2 sec, but differs from the human CNV in that, with the paradigm used, it is not well resolved by the imperative cue. Averaging evoked activity with the animal's motor response as reference point reveals another negative wave form which just precedes the motor response.


Evidence on the periodic mechanism in herpes simplex encephalitis. A case of herpes simplex encephalitis is described in which the EEG was recorded 5 times between the 5th day of illness and death on the 13th day. Repetitive discharges occurred at intervals of 3.5 or 2 sec; at first they were larger and earlier on the right side, finally on the left; on day 11 they could be earlier on either side and sometimes ceased briefly on either side. They were associated with jerks. It is argued that this periodicity cannot be explained by cortical isolation or a single pacemaker and hence favours a third possibility, a more or less diffuse disturbance of the time constant of excitability at the cellular level.


Nerve conduction velocity and refractory period as parameters of neurotoxicity. The maximum conduction velocity (MCV) and minimum refractory period (MRP) were measured in rat sacral ventral and dorsal roots, and saphenous nerves. At 32 degrees C values for MCV were 81.45, 73.96 and 49.12 m/sec for the ventral and dorsal roots, and saphenous nerves respectively, with corresponding temperature coefficients of 3.9, 3.25 and 2.14 m/sec/degrees C. Values of MRP at 32 degrees C for the 3 nerve preparations were 0.63, 0.52 and 0.47 msec, with an average temperature coefficient of 0.04 msec/degrees C at 32 degrees C.


The application of low-pass linear filters to evoked potential data: filtering without phase distortion. A linear filter, whose weights are based on the binomial coefficients, is described. It is shown that the phase distortion introduced by the filter is zero and the application of the filter to smoother EP records is illustrated.


Interpositus and fastigial unit activity during sleep and waking in the cat. Fine wire microelectrodes were used to record single unit activity from two of the intracerebellar nuclei, the interpositus and fastigius, during the sleep-waking cycle. The mean rates, interspike interval distributions and patterns of firing as revealed by autocorrelograms, were investigated. For each stage of the sleep-waking cycle and for as many units as possible the constancy of these measures was examined throughout several sleep cycles. Twenty-four interpositus units were recorded throughout at least one complete sleep-waking cycle. The firing rate of these units in paradoxical sleep without REM was equal to that in quiet wakefulness and greater than that occurring during slow wave sleep. The highest firing rates occurred during REM periods. Some interpositus units were found to have apparent eye movement related activity during REM periods but there was no correlation with waking eye movements. Eleven fastigial units were investigated throughout the sleep-waking cycle. At least half of these units were found to have rhythmic bursting activity during paradoxical sleep which did not appear to be tightly linked with REM periods. Fastigial and interpositus units were found to have slow shifts in their 'background' level of firing which occurred independently of the sleep-waking cycle. The results of these experiments are discussed in relation to the known inputs to the nuclei and in relation to previous studies on other groups of neurons during sleep and waking.


The significance of different sleep stages for the regulation of electrical brain activity in man. In 9 patients with implanted electrodes, the relationships between 13 deep subcortical structures and the premotor cortex were considered during different sleep stages, using both neuronal activities and slow electrical processes (including scalp EEG). Changes occurred asynchronously with a relative independence between the various brain structures during slow sleep. On the other hand, during REM sleep, all parameters reveal a high degree of synchronization of the activities. It is suggested that during slow wave sleep each of the separate rhythms display their own regulation, while in paradoxical sleep, it is their inter-relationships which are mainly controlled. The REM stage probably has some other function, associated with the fact that a neuronal population which has taken no part in any activity during wakefulness now reacts.


Automated detection of EEG artifacts during sleep: preprocessing for all-night spectral analysis. This paper describes a simple artifact detection algorithm which can be used when large amounts of EEG data are to be automatically processed via spectral analysis techniques in a general purpose digital computer, and visual inspection of each EEG epoch becomes an impossible task. The technique is based on a chi-square (chi(2)) goodness-of-fit test to a Gaussian distribution (CSQ), and it was applied to EEG epochs each 30 sec long. This test proved to be very sensitive to non-stationarities in the EEG amplitude distribution for a particular epoch, and it produced a large value for the chi(2) coefficient when an artifact was present. EEG epochs that gave rise to chi(2) coefficients of value larger than a heuristically determined minimum were discarded from further analysis. The above technique enabled efficient data reduction and reliable automatic off-line processing of 50 nights of sleep EEG via spectral techniques.


Role of the inferotemporal cortex in visual selective attention. Electrocortical recordings were made from monkeys performing in a multidimensional visual task. Wave forms dependent on the stimulus presented (irrespective of task required) were recorded immediately following the stimulus primarily from electrodes implanted in the striate and prestriate cortex. Wave forms dependent on the panel pressed (irrespective of the stimulus or of the task) were recorded especially from motor and post-central cortex, and to a lesser extent in anterior frontal cortex, always just prior to or following the time of the response. Wave forms dependent on the task as determined by the reinforcing contingencies (but independent of the particular stimulus presented or the particular panel pressed) were recorded primarily from the inferior temporal cortex, and rarely from prestriate and anterior frontal cortex. While task-related wave forms began to appear shortly after stimulus presentation, they became especially apparent around the time of the response. This response-linking increased in prominence as the subject achieved 90% proficiency in each task, only to drop off with overtraining. Further, the task-related wave form does not change as rapidly as does overt behavior when the reinforcement contingency is shifted from one stimulus dimension to another. The relevance of these results to an understanding of the process of selective attention is discussed.


Responses of lateral preoptic neurons in the rat to hypertonic sucrose and NaCl. Multiple-unit recordings were taken from the lateral preoptic region during a series of hypertonic and isotonic NaCl and sucrose intracarotid injections. Subjects were 11 hooded rats (8 males and 3 ovariectomized females) under urethane anesthesia. The data showed that under favorable cannulation conditions there were strong multiple-unit responses to hypertonic sucrose injections, and that under these conditions NaCl injections were not significantly more effective than sucrose injections. The possible bearing of these findings on hypotheses concerning central receptors for thirst is discussed.


The polarity inversion of scalp potentials evoked by upper and lower half-field stimulus patterns: latency or surface distribution differences? Evoked potentials to patterned stimulation of the upper and lower half of the visual field are generally inverted in polarity. Two conflicting proposals have been made to explain this effect, both based on surface distribution studies of pattern-reversal and/or pattern-onset VEPs. The first suggests that this polarity inversion is due to differences in surface distribution of corresponding components of constant latency; the second that it is due to differences in the latencies of peaks of similar surface distributions in the upper and lower half-field responses. Experimental evidence is here presented which supports the first explanation for the case of the pattern-onset VEPs. These results, which illustrate how different components in the same response can be identified from the selective adaptation effects of pre-exposure to outline patterns, show that there is no difference in latency of components of corresponding properties in the upper and lower half-field VEPs.


Cortical slow negative waves following non-paired stimuli: effects of modality, intensity and rate of stimulation. Using a simple stimulus counting task, negative after-waves following single (unpaired) stimuli were investigated under a variety of stimulus conditions. Responses were obtained to tones at 3 intensities, to light flashes at 3 intensities, and to tones at 3 rates of presentation. The acoustic stimuli led to a negative after-wave that peaked at frontal sites around 500 or 600 msec, and then trailed off to a more central scalp representation. This negative after-wave was increased in amplitude by slowing the rate of stimulus presentation. In comparison, no appreciable or sustained after-wave was elicited by visual stimuli. No significant effects in the negative after-wave were associated with intensity, either for visual or for acoustic stimuli. When analyzed by Principal Components Analysis, the negative after-waves were shown to comprise in all cases two underlying factors, although the factors contributed less to the total wave form for visual stimuli than for acoustic stimuli. Two interpretations for the negative after-wave were contrasted, one considering it to be an integral feature of the auditory evoked potential. A second interpretation, more compatible with the data obtained here, links the negative after-wave with non-specific activation processes.


The human amygdala. I. Electrophysiological responses to odorants. Nine patients had an amygdalotomy for attempted relief of intractable seizures and, as an aid for localization, recordings were made from the amygdala and later computer analyzed. The background activity is 3-18 c/sec; in response to odorants sinusoidal bursts are seen, synchronous with inspiration at 12-24 c/sec, but mainly 40-70 c/sec. Each odorant is associated with a given group of frequency components. However, for many different odorants the number of these components is restricted and the hypothesis is presented that odor discrimination in man's amygdala is not determined by which components are present, but the patterning of the components according to their amplitude. Odorants within the same odor class produced similar patterns of response. Epileptiform activity was recorded in two different forms, one in response to odors, similar in frequency to the slower bursts, except for sharper configurations, at times progressing to clinical seizures and the second as an insertion discharge from an advancing electrode, similar in frequency to the faster bursts. These latter data suggest that the repertoire of a given neural system or structure may be limited to specific frequencies but the configuration or patterning of these frequencies define the different states of the structure.


Differential changes of auditory nerve and brain stem short latency evoked potentials in the laboratory mouse. Subcollicular, volume-conducted auditory evoked potentials were obtained from the C57BL/6 laboratory mouse. Barbiturate-induced, whole body hypothermia was associated with a latency increase; between 37 and 31 degrees C, these values were 0.28, 0.48, 0.88 and 0.88 msec for PI-IV, respectively. The amplitude of the auditory nerve potential elicited by a 100 dB click doubled in amplitude between 16 and 42 days post partum. A progressive latency decrease, from PI to PIV, was observed during this age span. Acoustic stress produced differential changes within peripheral and central amplitude and latency measures, and these parameters were also sensitive to the method used to reduce body movement during the recording session. These changes were consistent with anatomical and physiological data from the auditory brain stem of other mammals.


Quantified electrographic scales on 10 pre-term healthy newborns followed up to 40-43 weeks of conceptional age by serial polygraphic recordings. Full-term newborns with seizures exhibit EEG patterns that appear reliable in predicting neurological outcome in almost 75% of the cases in a prospective investigation. These patterns are unreliable for over 25% of infants with seizures and even more so for term and pre-term babies with other complications. Observations that such newborns often show maturational delay in various physiological and/or behavioral parameters, raises the issue of whether indices other than specific EEG patterns might prove reliable in predicting prognosis. As a prerequisite for testing such an hypothesis in prospective investigations of term and pre-term babies with 'risk' factors other than seizures, it appears necessary to obtain quantification of these parameters in normals. Hence a longitudinal study was designed to obtain simultaneously 3 parameters in a control group of newborns born after 30-32 weeks of gestational age, followed with serial polygraphic recordings at weekly intervals until a conceptional age of 43 weeks. The selected parameters were purposefully restricted to three that can be obtained and measured easily in routine recordings without need for more complex instrumentation and analysis. These were: the percentage of interhemispheric synchrony between bursts of 'trace alternant'; the number of spindle-delta complexes ('brushes') during 5 min of REM and NREM sleep; and the concordance between various parameters during the same 5 min epochs. The range of these indices was thus established in a group of normal prematures followed prospectively. Establishing such normative data will allow greater inter-study reliability, and form the basis for other prospective studies of infants 'at risk' to investigate whether those that lag behind the norm in these indices when followed serially during the newborn period differ prognostically from those who recover and reach expected norms for conceptional age.


Evaluation of the Wiener filter applied to evoked EMG potentials. The application of the Wiener filter to the estimation of responses evoked in the tonic EMG activity of tibialis anterior by cutaneous electrical stimulation of the foot is described. The effectiveness of the filter is assessed using both experimental and simulated data by computing the mean square error between the actual evoked potential and ensemble average and Wiener filter estimates of it. The Wiener filter is shown to provide a better estimate of the final response than simple ensemble averaging. The improvement is most marked in cases where the signal-to-noise ratio is small, but the Wiener filter estimate is always better than the ensemble average estimate. It is concluded that Wiener filtering significantly reduces the number of responses required to obtain a good estimate of the evoked potential in this experimental situation.


REM sleep episodes during the Multple Sleep Latency Test in narcoleptic patients. Forty narcoleptic patients were given the Multiple Sleep Latency Test, consisting of 20 min opportunities to sleep offered at 10.00, 12.00, 14.00, 16.00 and 18.00 o'clock. Eleven patients had 2 episodes of REM sleep, 5 had 3, 11 had 4, and 13 had 5 before they were awakened. Fourteen control subjects given similar opportunities to sleep (reported in a companion article (Richardson et al. 1978)) had no REM sleep episodes. For the 10.00-18.00 o'clock opportunities respectively, there were 32, 29, 30, 28 and 27 REM sleep episodes. We conclude that this procedure can provide physicians with data useful in the diagnosis of narcolepsy.


A technique for separating endogenous from exogenous human cortical potentials. In a temporal discrimination paradigm, if signal duration is the stimulus dimension to be discriminated, the moment of subject's decision can be manipulated by varying the duration of the target stimulus. As stimulus duration is lengthened, task-contingent potentials related to the decision process can be separated from those evoked by stimulus onset. Analysis of individual evoked responses indicates that 3 task-contingent components, occurring at 220, 300 and 340 msec are precisely time-locked to the moment of decision.


Immunochemical studies of guinea pig progesterone-binding plasma protein. An antiserum specific for guinea pig progesterone-binding plasma protein (PBP) has been prepared. Using a very sensitive immunoenzymatic assay, PBP could be detected not only in pregnant guinea pig plasma (970 microgram/ml of plasma at day 40--60 of pregnancy), but also in the plasma of fetuses (2.77 micrograms/ml), umbilical arteries (1.79 micrograms/ml), umbilical vein (2.90 micrograms/ml), and in amniotic fluid (0.47 microgram/ml). The protein was also found in low concentration in the plasma of nonpregnant females (2.10 micrograms/ml) and males (1.56 micrograms/ml). The antiserum was used to search for immunological similarities between various steroid-binding proteins. No cross reaction was found with cavian or human corticosteroid-binding globulin and human sex steroid-binding plasma protein. There was no cross reaction between guinea pig PBP and PBP of other pregnant hystricomorphs (viscacha, degu, and coypu). Moreover, no evidence was found of an interaction between guinea pig uterine progesterone receptor and the anti-PBP antiserum.


Gonadotropin stimulation of porcine ovarian ornithine decarboxylase in vitro: the role of 3',5'-adenosine monophosphate. The role of cAMP as a mediator of gonadotropin stimulation of ovarian ornithine decarboxylase (ODC) activity was studied in granulosa cells isolated from small (1--2 mm) porcine ovarian follicles. These cells responded to both FSH and LH with significant increases in intracellular concentration of cAMP. At concentrations of gonadotropins which were saturating for the induction of ODC activity, FSH was a more potent stimulator of both cAMP production and ODC activity than LH. N,O'-Dibutyryl cAMP (1.0--10.0 mM) caused a dose-dependent stimulation of ODC activity which equaled the maximal effect of LH but was significantly less effective than the saturating dose of FSH. 8-Bromo-cAMP was more potent than N,O'-dibutyryl cAMP and as effective as FSH as an inducer of ODC activity. Addition of theophylline, a phosphodiesterase inhibitor, to the incubation medium resulted in a dose-dependent inhibition of ODC activity in both control and gonadotropin-stimulated cells. In contrast, 1-methyl,3-isobutyl xanthine, another phosphodiesterase inhibitor, potentiated effects of both submaximal and maximal effective doses of gonadotropins while producing no effect on basal ODC activity of these cells. The results of this study are consistent with the concept that cAMP can mediate gonadotropin stimulation of ODC in porcine granulosa cells. In addition, this study shows the importance of proper selection of cAMP analogs and phosphodiesterase inhibitors, and their concentration in studying such effects.


Induction of cyclic AMP phosphodiesterase by disaggregation of the multicellular complexes of Dictyostelium discoideum. When the multicellular complexes (slugs) of Dictyostelium discoideum were disaggregated, the activity of phosphodiesterase for adenosine 3':5'-monophosphate increased. The increase occurred first with the enzyme in the particulate fraction of the cell. Cytoplasmic and extracellular phosphodiesterase, as well as the macromolecular inhibitor for this enzyme, also increased at later times. The increase in these activities was blocked by inhibitors of protein and RNA synthesis. This indicates that the increase is not due to the activation of the enzyme, but due to its synthesis de novo, and that the synthesis is induced at the transcriptional level by the loss of cell contact. In the case of cellular phosphodiesterase, inhibitors of protein synthesis (cycloheximide and emetine) not only blocked the increase, but also caused a decrease in activity. The mechanism of such decrease was further investigated.


Effects of nicotine and hexamethonium on discharges in the stellate ganglion. The effects of nicotine and hexamethonium on postganglionic discharges elicited by tetanic preganglionic stimulation or muscarinic agonists were observed on the isolated hamster stellate ganglion. The amplitude and duration of the afterdischarges from tetanic preganglionic stimulation in hexamethonium (10-3 M) were smaller than the amplitude and duration of the afterdischarges in nicotine (10-3 M). Also, hexamethonium decreased the amplitude and duration of the afterdischarges from repetitive stimulation in the presence of nicotine. The mechanism of these effects was explored. After application of nicotine for 30 min, the discharges from McN-A-343, a muscarinic agonist, were the same as before the nicotine. Hexamethonium did not reverse the block of the single evoked potential by nicotine. The potentials during a train in the presence of hexamethonium plus atropine were the same as the potentials during a train in the presence of nicotine plus atropine. Hexamethonium did depress the McN-A-343 discharges in the presence of nicotine and also in the control solution. These results indicate that hexamethonium has a direct depressant effect on the muscarinic synaptic membrane.


Internal synthesis of p23,30 by several lymphoid malignancies. The aim of this study was to prove the internal synthesis of p23,30 antigen (HLA-D related determinant) on human leukemias and lymphomas on which it has been detected with complement-dependent cytotoxic assays. Murine Ia antigens similar to p23,30 antigen are found on many subsets of cells in the mouse (B lymphocytes, macrophages, allogeneically activated T lymphocytes) and on intercellularly transferred immunoregulatory molecules, which may be adsorbed to other cells. The question exists whether the p23,30 antigen, which occurs on a wide range of human leukemias, is internally synthesized by these tumors or, in some instances, is synthesized by normal lymphocytes and is adsorbed to the leukemic cells. The expression of p23,30 antigen on a limited series of human leukemias and lymphomas was detected by a complement dependent, cytotoxicity assay. The internal synthesis of p23,30 antigen and p44,12 (HLA-A and -B antigens and beta2-microglobulin) was confirmed by immunoprecipitation and these antigens from [35S]methionine labeled, detergent solubilized membranes of tumor cells. In each instance, the synthesis of p23,30 antigen by the malignant cells was confirmed. The distribution of p23,30 antigen (and 1a antigen) on subsets of normal cells and in immunoregulatory molecules was reviewed. In view of these findings, the role of p23,30 antigen in the diagnosis of subsets of human hematologic malignancies was reconsidered.


Morphometric investigations on endocrine glands. V. Changes in the testes of Wistar rats after application of chlormadinone acetate and norethisterone acetate. In growth inhibiting tests of rat testes the tubular diameter and, somewhat less sensitive, the nuclear volume of the Leydig cell follow the weight changes. Chlormadinone acetate shows antigonadotropic activity only at high doses and a dose-dependent antiandrogenic activity. The dose-weight curve under norethisterone acetate has a minimum at 4.2 mg/rat/14 days; higher doses seem to be accompanied by androgenic efficiency.


[A steady state polyacrylamide gel electrophoresis for the determination of testosterone-estradiol binding globulin (author's transl)]. Testosterone-estradiol binding globulin (TeBG) is known to change in various endocrinological environments such as estrogen administration, pregnancy and aging. Several methods, including dextran coated charcoal, equilibrium dialysis and ammonium sulfate precipitation, were used to measure the binding capacity of TeBG, but these were not simple and accurate. We therefore measured TeBG levels in human serum by means of a steady state polyacrylamide gel electrophoresis and found that this method was simple and accurate for the determination of the binding capacity of TeBG. The value of TeBG in normal adults (27 approximately 32 years old) was 3.88 +/- 0.45 x 10(-8) Mol and in patients with benign prostatic hypertrophy the value was high (5.49 +/- 1.35 x 10(-8) Mol compared to that of normal adults.


Failure of immune serum globulin to prevent hepatitis B virus infection in infants born to HBsAg-positive mothers. Twelve infants, born to mothers with hepatitis B virus infection, were inoculated within 7 days of birth with immune serum globulin containing antibody to hepatitis B surface antigen (HBsAg) titers of 1:32 to 1:64 as measured by passive hemagglutination. Six of nine infants (66.7%) born to HBsAg-positive carrier mothers became HBsAg-positive within 3 mo of age. In addition, two of three treated infants born to mothers with acute hepatitis B during the delivery period also developed HBsAg. The hepatitis e antigen was detected in four of five carrier mothers and in two mothers with acute hepatitis, whose infants subsequently became HBsAg positive. In addition, hepatitis B-specific DNA polymerase activity was detected in the seven HBsAg-positive mothers who transmitted the virus to their infants. All eight infants have remained persistently HBsAg positive. Thus, the immune serum globulin containing low-titer antibody to HBsAg is not protective when given to infants born to HBsAg carrier mothers or to mothers with acute hepatitis B during the delivery period.


[Classification of non-Hodgkin's lymphomas: facts and perspectives]. Previous classifications of malignant non-Hodgkin lymphomas (nHL) are based on morphological and histological-cytological findings. The rapid development of immunology, the application of immunocytological, cytochemical and electromicroscopic methods led to new classifications of nHL based on immunology. They are founded on the idea that the prevailing cells of nHL correspond to a cell type from the differentiation line of lymphocytes. Thus, a distinction can be made between malignant lymphomas (ml) of stem, B and T-cells; the more frequently occurring lymphomas originating from B-cells are represented by chronic lymphatic leukaemia originating from B1-cells, by ml of follicular genesis (centrocytes, centroblastic-centrocytic ml, centroblasts) and by ml synthesizing and releasing immunoglobulins (immunocytoma, immunoblastoma). The type formerly called "reticulosarcoma" (histocytic ml) could be identified to be an immature, Ig synthesizing ml (immunoblastloma) in most cases. The new classification enables a more extensive differentiation and a better prognostic statement to be made than previous ones. The new units of disease can be diagnosed in most cases with light microscopic routine methods, although borderline cases and unclassifiable ml may occur.


[Mitotic index in the bone marrow of children during the clinical course of acute lymphoblastic leukemia (ALL)]. From 31 children with acute lymphoblastic leukaemia the mitosis index in the bone-marrow was determined before the onset of therapy and during the clinical progress. Initially, the mean white mitosis index lay with 3.4% below that of the normal test persons, it rose significantly in the hematologic full remission and showed a decreasing tendency with a great range of dispersion in the recidive. The most lowered mitosis index was found in the final stage. Strong shifts in the kariologic distributions make a remaining in the prophase of the mitosis evident. The influence of polychemotherapy on the mitosis index and the phases of mitosis is discussed. Correlations between the mitosis index and clinical as well as paraclinical parameters were only to be found with respect to granulocytes and lymphoblasts. The considerable ranges of fluctuations of the mitosis index and the lack of congruity with the clinical progress of ALL allow no ensured assertions to be made for the single patient. It seems to be important in eosinophilia and in leukaemoid reactions.


[Sézary syndrome. A case report].

[Sézary syndrome. A case report]. This paper describes a patient who exhibits all signs of Sézary-syndrome. During the 15 years of sickness, the patient has developed a generalised erythrodermia, skin infiltrations, hyperkeratosis on the palms and the foot soles, changes in pigmentation, and generalised swelling of the lymph nodes. Two modifications of Sézary's cells have been found in the peripheral blood and the specific skin infiltrations by both light and electron microscopy. The serum contains large amounts of a macrophage migration inhibitory factor. The illness does not respond to chemotherapy.


[Quantitative behavior of the B-lymphocyte subpopulation in peripheral blood of children of different ages]. By means of a direct immunofluorescence technique the numbers of B-lymphocytes in the peripheral blood were determined in healthy children of different age with anti-human IgG serum and anti-human globulin. Their small number in new borns and a marked increase in infancy were significant. In addition, the paper contains extensive methodical data.


Group systems of serum proteins and erythrocyte enzymes in malignant lymphoma. Four group systems of serum proteins (Hp, Gc, Gm, Km) and five group systems of erythrocyte enzymes (AP, PGM1, GPT, AK, EsD) were determined in 63 patients with malignant lymphoma. Statistical analysis of the distribution of the above mentioned systems in patients and Polish population samples did not reveal any significant differences, which points to the lack of any correlation between the disease and the group systems under examination.


The lymphocyte cytochemical equipment and serum immunoglobulins in patients with precancerous states of the larynx. In 24 men with precancerous states of the larynx, i.e. leukoplakia, papillomas and pachydermia, the peripheral blood lymphocytes were cytochemically stained for N-acetyl-beta-glucosaminidase, beta-glucuronidase, acid phosphatase and glycogen (PAS reaction). The results were expressed in terms of the absolute counts of enzyme- (or compound-) positive cells. The serum immunoglobulin IgG, IgA and IgM levels were also determined by Mancini's method. The results obtained were compared with those in 20 healthy men aged 20 to 30 years. It was found that the patients exhibited elevated numbers of N-acetyl-beta-glucosaminidase- and beta-glucuronidase-positive lymphocytes. A characteristic feature was an increase in the absolute counts of lymphocytes with diffuse and granular-diffuse types of cytochemical reaction for all enzymes studied. The number of cells with the granular type of enzymatic reaction (intact enzyme-positive lysosomes) was significantly diminished. These cytochemical alterations were accompanied by a significant increase in the serum IgA level. These results are discussed with reference to the lymphoid system response to tissues with precancerous lesions of the larynx.


The cytochemistry of rabbit peripheral blood leukocytes. The authors report on their investigations of the differentiation between neutrophilic and eosinophilic granulocytes of the peripheral blood in healthy rabbits. By varying the pH-value and the incubation time it is possible to achieve a reliable differentiation between neutrophilic and eosinophilic granulocytes of the rabbit by means of 6 different cytochemical methods which could only be made incompletely with the routine methods used up till now (Eosin-, Giemsa-, Wright's-colouring etc.). Moreover, monocytes and lymphocytes can also be identified reliably.


Role of L-thyroxine, hydrocortisone and progesterone in regulation of blood volume in a fresh-water air-breathing fish, Clarias batrachus (Linn.). Effect of hormones on the blood volume of Clarias batrachus were ascertained and it was observed L-thyroxine and progesterone decrease, while hydrocortisone increases the total and relative blood volumes. The ratio of plasma to corpuscular volume is increased by the intraperitoneal injection of L-thyroxine, hydrocortisone and progesterone (higher dose only), thus indicating the regulation of blood volume under the complex control of homeostasis.


Effect of hormones and pharmacological drugs on the blood of Anabas testudineus (Bloch). Effect of different gonadal steroids, hydrocortisone, catecholamines and antithyroidal agent on certain haematological parameters in an air breathing fish, Anabas testudineus (Bloch) commonly known as climbing perch, have been made. The treatment of hydrocortisone, adrenaline, noradrenaline and testosterone brings about a significant increase in RBC counts (polycythemia), Hb content and PCV values as compared to control which were subjected to the treatment of normal saline. Males were more susceptible to the treatment of hydrocortisone, catecholamines than females. Ovocyclin, Duogynon forte and Fertyl treatments bring about anaemia in this fish, i.e. a decrease in RBC number, Hb content and PCV was clearly marked out. Females were more susceptible to Fertyl than males. Lutocyclin failed to bring any significant changes in these blood parameters. Though the treatment of thiouracil could not bring about significant changes in different blood parameters, a tendency of increase in RBC counts, Hb content and PCV was observed. The absolute values for different blood parameters have also been calculated. Possible reason and the mechanism of such changes in different blood parameters are discussed in detail.


Glutaraldehyde mediated echinocyte/discocyte transformation is Ca2+ dependent. Echinocytes, which were produced from freshly banked blood by repeated washes in phosphate buffered saline, undergo a transformation to the discoid shape within less than 30 seconds of incubation in isotonic 0.05% glutaraldehyde pH 7.4. This echinocyte/discocyte transformation is not associated with a change of cell volume or critical hemolysis volume although a slight decrease of cellular deformability and a 4-8 fold increase of K+ efflux within 1 hour after glutaraldehyde incubation provide evidence of the fixative's attack on the cell membrane. Trypsination prior to the incubation in isotonic glutaraldehyde could not inhibit the shape change. Hypertonic glutaraldehyde solutions partially prevent the E/D transformation with regard to both the osmolarity of the medium and the permeability of the cell membrane. The glutaraldehyde stimulated transformation is entirely inhibited in the presence of a chelating agent the efficiency of which is overcome by addition of a more-than-equivalent amount of Ca2+. The mutual action of either agent is discussed, however, the mechanism of the phenomenon remains unclear.


[Modification of sialic acid determination in erythrocytes]. Various chromophores impair the evaluation of N-acetyl-neuraminic acid (NANA) of red blood cells by means of the thiobarbituric acid method according to WARREN. Therefore, a time consuming isolation of erythrocyte membranes is essential in order to avoid these shortcomings. The procedure was simplified by fixation with glutaraldehyde of washed red blood cells.


A new simple method for determination of erythrocyte filtrability. A new simple filtration technique designed for measuring red cell filtrability in the routine laboratory use was developed. The suspension of the whole blood in saline (1:20,000 dilution) was processed on the Sartorius filter membranes, pore size 8 micron. The percentage of passed erythrocytes indicating red cell filtrability was determined. The suitability and perspective applicability of this method for studying various hematological disorders is proposed.


Decreased thrombotic mortality and morbidity after valvular cardiac surgery as a consequence of adequate coumarin anticoagulation. A retrospective study in 516 patients. Two almost identical groups of 258 patients each who underwent surgery for the substitution of one or more than one cardiac valve were taken into consideration. After surgery all patients, with the exception of those who received a porcine allograph, were given coumarin drugs for 1 year for thrombosis prophylaxis. In the first group of patients operated in 1972 and 1973 the laboratory follow-up and the therapeutic range were neither constant nor uniform. In the case of the second group, which included patients operated in 1974 and 1975, adequate laboratory follow-up and constantly adequate therapeutic ranges were always kept. This retrospective study has shown a statistically significant decrease in mortality due to thromboembolism and to unknown causes in the second group of patients (x2 = 8,313, "p" less than 0,0025). A decrease of the non-lethal embolic complications was also noted in the second group of patients (x2 = 6,813, "p" less than 0,0025). On the other hand, bleeding complications were greater in the second group of patients (x2 = 3,485, "p" less than 0,05) and this was interpreted as secondary to the higher anticoagulation administered to the latter patients. The results indicate that an adequate coumarin anticoagulation seems able to decrease the incidence of lethal or non-lethal thromboembolic complications after cardiac valve replacement significantly.


The economic side of blood banking and plasma fractionation. While cellular components have a relatively short half-life, must be necessarily administered group-specifically and kept available on a round-the-clock basis, all of which add significantly to the basic cost per unit, products isolated from plasma are not handicapped by these disadvantages. Another important advantage for the production of plasma components lies in the fact that the raw material may be also collected through plasmapheresis, a process which allows the collection of significantly greater amounts of plasma from one donor as compared to conventional whole blood collection. Quite understandably, the maintenance of whole blood and cellular component supplies has been left to national and/or non-profit organizations, while commercial firms run a profitable business with the production and distribution of plasma fractions. The method for the selective isolation of plasma fractions developed in our blood transfusion service solves the high cost problems involved in conventional fractionation methods but does not solve the ethical and economical problem related to discarding precious unused raw material.


[Granulocyte kinetics with radioactive diisopropylfluorophosphate (DF32P) and radiochrome (51Cr)]. Determining granulocyte kinetics with DF32P allows various parameters to be gained during the in-vitro marking, such as the total blood granulocyte pool, circulating granulocyte pool, marginal granulocyte pool, daily granulocyte exchange rate and half decay period of granulocytes. The half decay period of granulocytes, bone-marrow reserve in myelocytes, metamyelocytes and band cells as well as polymorphonuclear neutrophils can be determined by in-vitro marking, with DF32P being intravenously injected. The combination of both procedures with DF32P will reveal the half decay period, pool sizes and exchange rates of the proliferating myelocyte compartiment in bone-marrow and mature blood granulocytes. If 51Cr is used for determining granulocyte kinetics the surface activities of various organs, such as heart, liver, spleen, and lungs, can mainly be determined in addition to the half-life of leucocytes, indicating the degradation or storage of cells in certain areas of the body. In addition to normal values those findings are principally presented which were obtained with in-vitro marking by DF32P and 51Cr in chronic myeloid leukaemia, osteomyelofibrosis or osteomyelosclerosis respectively and in hypersplenism.


Immunostimulatory effect of zinc in patients with acute lymphoblastic leukemia. The studies have been performed in 27 ALL patients, aged 4 to 21 years, during their first remission induced by the Memphis Center therapeutic schedule (New Eng. J. Med., 1974, 5, 1230). The onset of therapy with zinc took place immediately after cessation of remission supporting treatment (Vincristine, Prednison, intrathecally administered Methotrexate). During the seven days period of investigation all the patients studied were simultaneously treated with 6-Mercaptopurin. The patients were divided into two groups. The first group (15 subjects) received 0.02 mg Zn per kg of body weight (in the form of zinc sulphate prepared by Kutno Pharmaceutical Establishment POLFA). The second group (12 subjects) received placebo. The zinc administration resulted in statistically significant (p less than 0.05) increase in the TEa5' percentage and the increase of both percentage and absolute number of TEt60', whereas the serum total gamma globulins, as well as IgG, IgA and IgM levels exhibited no alterations. The examined indices of both humoral and cellular immunity did not change in the control group. The zinc therapy was not accompanied by side effects, except the slight decrease in the granulocyte count. The final opinion concerning the clinical use of zinc as cellular immunity stimulatory agent in the ALL therapy deserves further studies.


Cultivation of fresh and frozen mouse bone marrow. - Application of methyl cellulose. The culture of cells of both fresh and frozen mouse bone marrow on methyl cellulose (MC) was approached. We used 1.5%-concentration of MC and proved the stem cells of fresh and frozen mouse bone marrow to proliferate and form haemopoietic colonies on MC. We established that the freezing process did not significantly decrease the proliferative capacity of CFU-C stem cells.


[Critical remarks on the monocyte test]. The application of the monocyte test in the diagnostics of drug allergy is considered from a critical point of view. Further hospital application of the test and its evaluation are discussed.


[Coincidence of chronic lymphadenosis and lymphogranulomatosis]. A case is reported on the joint presence of chronic lymphatic leukaemia and lymphogranulomatosis in a 58 years old patient. 5 years after diagnosis and therapy of lymphadenosis a lymphogranulomatosis existing in addition to lymphadenosis was discovered by autopsy. Previously a change of symptoms had occurred. This became evident in the regression of palpable lymph node swellings as well as leukocytosis and lymphocytosis, in a change of the differential blood picture and the presence of periodic subfebrile temperatures. Problems connected with these double neoplasias are briefly outlined and discussed.


[A plasmacytoma with changing paraproteinemia]. A disease on myeloma is described which originally showed an IgA paraproteinaemia. 3 years later a paraprotein of IgM-typ could be identified by the immunoelectrophoresis. Clinical progress and examination of the bone marrow revealed the unequivocal diagnosis of multiple myeloma. In spite of IgM paraproteinaemia the formation of Waldenströms' disease did not occur.