Comparative study of different insulin regimens in management of diabetic ketoacidosis. 36 patients in severe diabetic ketoacidosis were studied prospectively. All patients were treated with small doses of insulin, the first 18 by the intramuscular route and the remainder by continuous intravenous infusion. These patients were compared with 25 ketoacidotic patients who were treated with large intravenous boluses of insulin. With the intramuscular regimen there was a slower fall of blood-sugar than with intravenous insulin administered either in boluses or as a continuous infusion. The acidosis took longer to correct when small doses of insulin were used. In practice a long time interval between correction of the hyperglycaemia and correction of the acidosis may be a problem when the continuous infusion of insulin is used. Potassium requirements during treatment were identical (30-40 mmol/l fluid infused) and independent of the insulin regimen. However, small doses of insulin led to a poor retention of potassium. In the management of diabetic ketoacidosis the mode of administration of insulin is of limited significance, and small doses, although effective in most cases, are not clearly superior to conventional treatment with large doses.


Combination therapy for advanced resistant Hodgkin's disease. Thirty-nine patients with advanced Hodgkin's disease (H.D.) resistant to standard combinations of cytotoxic drugs were treated with C.C.N.U., vinblastine and bleomycin (C.V.B.). Ten patients (25%) achieved complete remission (C.R.) and seven of these are still in C.R. at the time of this report. A further 23 patients (59%) achieved partial remissions. The overall response-rate was thus 85%. C.C.N.U. caused less nausea and vomiting than that usually associated with nitrogen mustard. C.C.N.U. and vinblastine caused myelosuppression, but C.V.B. could be administered every 4 weeks to 21 (54%) of the 39 patients. A combination of cytotoxic drugs such as C.V.B. may be preferable to single agents in the treatment of patients with advanced resistant H.D.


Adherence and blood-pressure response to hypertension treatment. 2 years of experience in the first phase of a large cooperative national hypertension programme yielded data indicating that a good level of patient adherence can be achieved and that satisfactory blood-pressure control can be maintained long term. For 116 participants, all employed persons, dropout in the first year was 20% but only 3% dropped out  in the second year. At the second annual examination, 82% of those still in the programme had diastolic pressures under 90 mm Hg, with an average reduction of 18 mm Hg. Thus, nearly two-thirds (64%) of all patients originally enrolled were both active and with normal levels of blood-pressure after two years. Only 18% of active programme-treated patients had diastolic pressure 90 mm Hg or higher at the second anniversary in contrast with 33% of patients referred to their own doctors. In programme-treated patients, standard medication was used; diuretics effectively lowered blood-pressure in a third of patients, and diuretics plus reserpine were effective for another 20%. Special features of the programme included assistance to physicians by health counsellor therapists. Methods for achieving a high adherence-rate and satisfactory blood-pressure control probably have wider applicability.


Intentional radioiodine ablation in Graves' disease. In a series of 50 cases of Graves' disease qualifying for therapy with radioiodine, "ablative" doses of radioiodine resulted in hypothyroidism in 92% of cases at six months. Hypothyroidism was treated by thyroxine or tri-iodothyronine replacement therapy. Rapid responses and predictable outcome were regarded as important advantages. Intentional ablation is particularly convenient in situations where clinical and biochemical follow-up may be difficult. Further follow-up studies are needed to determine whether ablative therapy is the treatment of choice in all cases of Graves' disease.


Measurement of thyroid-stimulating hormone in dried blood spot. Blood T.S.H. (thyroid-stimulating hormone) was measured by radioimmunoassay in dried blood spotted onto filter-paper and obtained during screening of the newborn for metabolic disorders. By this method, the detection limit for blood T.S.H. was 5--10 muU/ml, which is the approximate upper limit of normal for blood T.S.H. T.S.H. values obtained on dried blood correlated well with those obtained on serum from the same subjects. Duplication of the assay in a single sample is not necessary. This method picked up a case of primary hypothyroidism in a four-year-old girl with symptoms. Since the technique is simple and sensitive enough for the detection of hypothyroidism, it could be valuable in mass-screening for congenital hypothyroidism.


Natural course of symptomless autoimmune thyroiditis. Out of 18 subjects with symptomless autoimmune thyroiditis (S.A.T.) 5 (28%) became hypothyroid within 4 to 39 months of observation, whereas 13 (72%) remained euthyroid up to atleast 28 to 50 months. In all those who developed hypothyroidism the basal serum level of thyroid-stimulating hormone (T.S.H.) was already initially above normal (normal range 1-6--6-9 muU/ml) and 4 had markedly elevated concentrations (less than 19 muU/ml). All those subjects developing hypothyroidism also had initially an exaggerated response to thyrotropin-releasing hormone (T.R.H.) (upper normal limit, delta T.S.H. 30 muU/ml), and in 4 the response was much exaggerated (delta T.S.H. less than 70 muU/ml). In 3 of these subjects the basal T.S.H. and the response to T.R.H. were reassessed before starting the substitution therapy and in all there was a further increase in both values. The basal serum T.S.H. was initially also slightly increased in 2 and the response to T.R.H. slightly above normal in 4 subjects who remained euthyroid. The basal T.S.H. level became normal in both cases with elevated values; and the response to T.R.H. declined to a normal level in 3 of the latter 4 subjects, but showed a further increase in 1. The thyroglobulin antibody (TgA) titres were initially significantly elevated in 15 subjects and the thyroid microsomal antibodies (MsA) in 1. The TgA titres decreased during the observation period in all but 1 subject and a similar trend was observed as regards the MsA titres. It is concluded that within a few years of observation a substantial number of subjects with S.A.T. will be hypothyroid. A definitively increased basal serum T.S.H. level and a markedly exaggerated response to T.R.H. in the symptomless stage of the disease is connected with a high risk of late hypothyroidism.


Smooth-muscle antibodies in infertility. Sera from 77 infertile women and 77 post-partum controls were examined for autoantibodies by indirect immunogluorescence. 27 (35%) of the infertile group had smooth-muscle antibody compared with 2 (3%) controls. Significantly more infertile patients had anti-nuclear factor, and reticulin antibodies were also more commonly found in this group. The differences for organ-specific autoantibodies between the two groups were not significant. There was no correlation between the occurrence of smooth-muscle antibodies and immunofluorescent anti-sperm antibodies. The high frequency of smooth-muscle antibodies in infertile women requires further study to determine whether an underlying viral infection is involved.


Preparation of the bowel by whole-gut irrigation. Experience with whole-gut irrigation as a method of bowel preparation in eightyone patients is described. The mean (+/-S.D.) weight-gain during irrigation was 1-9 +/- 0-8 kg; potassium losses in the effluent after one hour (4-0 +/- 2-5 g) were not significantly altered by adding potassium chloride to the irrigant. Eight irrigations were unsatisfactory, three being due to unrecognised obstructive neoplasms. The method provided excellent preparation for colonoscopy and large-bowel resection with anastomosis and was well tolerated by the patients.


Intraportal insulin protects from the liver injury of portacaval shunt in dogs. 4 days after portacaval shunt, the livers of normal dogs had pronounced atrophy and other structural abnormalities. These changes were greatly reduced in the left liver lobes, but not in the right, by a constant infusion to the left portal vein of insulin in non-hypoglycaemic doses. These experimental findings should have implications in clinical medicine.


Immunotherapy of non-clinical vaginal cancer. In six women who had had a hysterectomy and three or more vaginal smears positive for malignant cells but no symptoms, a course of immunotherapy, based on delayed hypersensitivity reaction to dinitrochlorobenzene, was tried. Repeat smears done every three months have all been normal, and the patients were well when last seen 2-35 months after treatment.


Functional subdivision of islets of Langerhans and possible role of D cells. Immunocytochemical examination of the islets of Langerhans in various animal species, including man, indicates that insulin-producing cells (B cells), glucagon-producing cells (A cells), and cells producing somatostatin or a somatostatin-like peptide (D cells) are not randomly arranged within the islet. Whenever A cells are found in the islet--i.e., mostly in its peripheral part--they are accompanied by D cells. However, most B cells, which occupy a central position, are in contact only with other B cells. In view of the inhibitory effect of somatostatin on both insulin and glucagon secretion, it is suggested that the arrangement of A, B and D cells is important to the normal and pathological functioning of the islet.


Impact of change to early diagnosis and surgery in major upper gastrointestinal bleeding. The diagnostic, operation, and mortality rates in two series of patients with major upper gastrointestinal bleeding have been compared. In one (1968-69) patients were admitted to medical wards, radiology was the only investigation, and few were operated on. In 1972-74 a more vigorous policy was adopted with investigation by both radiology and endoscopy, more (and earlier) surgery, and intensive care by a surgical team. Failure to reach a diagnosis fell from 38-9% to 5-7%; the operation-rate increased; transfusion requirements decreased; and mortality in both operated (47% v. 11%) and non-operated (17% v. 8%) fell. A policy of early and vigorous investigation and management by a surgical team is urged for patients presenting with severe bleeding from the duodenum, stomach, or oesophagus.


Changing patterns of bacterial resistance in relation to prophylactic use of cephaloridine and therapeutic use of ampicillin. Antibiotic sensitivities of 15302 organisms of common pathogenic species isolated in one hospital pathology department in 1971 and 1974 have been studied. Resistance to cephaloridine did not change materially. The proportion of strains resistant to ampicillin did, however, increase, and this is attributed to the widespread therapeutic use of the antibiotic. Cephaloridine, on the other hand, was largely used in the hospital as a single-dose, intra-incisional prophylactic against surgical wound sepsis.


[On the absorption of decongestive nose-drops-investigation with a radioactive labelled imidazoline-derivate (author's transl)]. In animal experiments, the absorption of a nasal decongestive substance after application to the nasal mucosa was studied. After an exposition-time of 60 minutes, about 1% of the substance applied could be measured in the total blood volume. An incorporation of the substance into proteins of the nasal mucosa could not be found. By means of frozen-section-autoradiography, the way of the applied decongestive substance through the mucosa could be investigated. The reasons for the relative small absorption rate are discussed. The chemical structure and the vasoconstrictory effect probably are reliable for that.


Localization of laryngeal motor neurons in the kitten. In a series of 12 newborn kittens, horseradish peroxidase (HRP) was used to trace retrograde axoplasmic flow in the motor neurons to laryngeal muscles. The animals were sacrificed 24 hours after injection of HRP into a specific laryngeal muscle, and the brain stems were stained for peroxidase. This clear-cut colorimetric method permitted the localization of the motor neurons in two nuclei of the ipsilateral brain stem. These are the nucleus ambiguus and the retrofacial nucleus. The primary source of laryngeal motor supply is the nucleus ambiguus. All the laryngeal muscles were represented here in two divisions. Adductor neurons were located in the dorsal division and were more loosely arranged in the lateral reticular formation. The abductor neurons of the posterior cricoarytenoid muscle formed the compact ventral division of the nucleus ambiguus and were fewer in number than adductor neurons by a factor of four to one. Since the expiratory and inspiratory centers are also located dorsally and ventrally in the brain stem reticular formation, the motor cells of the nucleus ambiguus are conveniently arranged to receive their afferent input. This arrangement is probably the result of phylogenetic development of abductor laryngeal function and pulmonary function in lower forms. A second source of laryngeal innervation is the retrofacial nucleus. This small nucleus is situated rostral to the nucleus ambiguus and is made up of small and medium-sized neurons of at least two types. Only the cricothyroid (CT) and posterior cricoarytenoid (PCA) muscles were shown to have significant innervation from this nucleus. The CT neurons were located peripherally while the PCA cells occupied the central portion of the nucleus. The functional significance of this nucleus is unknown, but it is suggested that it may have something to do with the various types of muscle units that have been demonstrated physiologically in the CT and PCA muscles.


Antigenic relationship between the surface antigens of avian and equine influenze viruses. Influenza virus Equine 1 (A/equine/Prague/56) has a hemagglutinin which is antigenically related to the hemagglutinin of fowl plague virus strain Rostock (FPV) and a neuraminidase which cross-reacts with the enzyme of virus N (A/chick/Germany/49). After a single injection of chickens with Equine 1 virus no hemagglutination inhibiting (HI) and neutralizing antibodies against FPV can be demonstrated, although the birds are fully protected against a lethal dose of FPV. HI and neutralizing antibodies against FPV appear after a second injection of Equine 1 virus several weeks after the first one. Liberation of newly sunthesized FPV from the host cell is ingibited by antibodies cross-reacting with any antigen of virus surface.


DNA-synthesis on giant nuclear RNA by AMV DNA polymerase. The reverse transcription of pre-mRNA isolated from rat liver or mouse Ehrlich ascites carcinoma cells with the aid of hot phenol fractionation technique is described. Pre-mRNA isolated at 85 degrees C is a more active template than the 65 degree C fraction. The addition of oligo(dT) as a primer strongly stimulated the template activity of the 65 degree C fraction. The size of product corresponds to a sedimentation value of 7 S as measured in alkaline sucrose gradient and is essentially less than the size of template.


Reverse transcriptase in leukocytes of leukemic patients in remission. A cytoplasmic particulate fraction from human leukemic cells has been shown to contain reverse transcriptase and its associated high-molecular weight RHA template. We attempted to detect the reverse-transcriptase-template complex in morphologically normal peripheral blood leukocytes from patients with acute leukemia in complete remission. Our assay system consisted of a velocity glycerol gradient and cesium sulfate equilibrium gradient analysis of the  endogenous reverse transcriptase reaction product. Three of nine patients in remission had positive reactions determined by glycerol gradient analysis, and eight of 10 patients in remission had positive reactions by cesium sulfate gradient analysis. We were unable to detect the template complex in leukocytes of normal persons. Thus, normal-appearing leukocytes in the peripheral blood of some leukemia patients in remission seem to retain a number of biochemical characteristics, possibly viral related, associated with leukemic cells.


Surface markers and prognostic factors in acute lymphoblastic leukemia. We investigated the surface markers on lymphoblasts from 37 patients with acute lymphoblastic leukemia. Spontaneous rosette formation with sheep erythrocytes (E rosettes) identified T cells and the presence of surface immunoglobulin identified B cells. Eight patients had T-marker lymphoblasts; 28 had no markers (null lymphoblasts), and one patient had B-marker lymphoblasts. The eight patients with T-marker acute lymphoblastic leukemia had massive leukemic infiltration, frequently a mediastinal mass, and a poor prognosis. The T-marker lymphoblasts had a weak or negative periodic acid-Schiff reaction and a very low antigenic stimulation to allogeneic lymphocytes. The association of T-marker lymphoblasts and some of the conventional poor-risk factors appears to be reliable in predicting a poor prognosis.


Intraaxonal and extraaxonal transport of 125I-tetanus toxin in early local tetanus. The distribution of radioactivity in the sciatic nerve, the spinal ganglia, the ventral roots and the spinal cord was studied by means of histoautoradiography after injection of 125I-labelled tetanus toxin into gastrocnemius muscles of cats. In the sciatic nerve the major part of the radioactivity was found in the epineurium, but some axons also contained radioactivity. In the ventral root the radioactivity was strictly confined to a few axons; no radioactivity was found in other parts of the ventral root. In the spinal cord the radioactivity was confined to a few motoneurones where it was found in the soma as well as in the dendrites. Transient cooling of the ventral roots prevented the ascent of radioactivity into the spinal cord. Colchicine and vinblastine, after local application to the sciatic nerve, reduced the amount of radioactivity found in the ventral roots and in the spinal cord. However, the same effect was also obtained but to a lesser degree with lumicolchicine. It is concluded that the intraaxonal compartment is involved in the neural ascent of tetanus toxin into the spinal cord.


Biodegradation of alpha-hexachlorocyclohexane. VI. The cechlorination of alpha-hexachlorocyclohexane by microsomes and cytosol of rat liver. Alpha-hexachlorocyclohexane (alpha-HCH) is dechlorinated by enzymes contained in rat liver cytosol and microsomes. An evidence was obtained that in the cytosol there are two alpha-HCH dechlorinating enzymes at least; one operates only in the presence of reduced glutathione (GSH) and catalyzes dechlorinations associated with the formation of another hydrophilic product. This product is probably a conjugate of the alpha-HCH-residue with GSH. The other cytoplasmic alpha-HCH-dechlorinase requires no additions. The microsomes, too, contain two alpha-HC dechlorinases at least: one is stimulated by GSH, the other by NADPH.


Demonstration of synthesis of beta-trace protein in different tissues of squirrel monkey. The sites of synthesis of the low molecular weight beta-trace protein, present in a seven times higher concentration in normal human CSF than in normal human serum, have been studied by means of a radioactive immunoprecipitation method. Adult squirrel monkey tissues were cultured in Eagle's minimum essential medium in the presence of 14C-labelled valine, threonine and leucine for 24 hours. Synthesis could be demonstrated in cultures of white CNS matter, whereas cultures of grey CNS matter, peripheral nerve, skeletal muscle, kidney and ovary did not show any signs of synthesis. Some cultures of spinal cord, basal ganglia, genital organs except ovary, and liver showed a probable synthesis of beta-trace protein. By means of autoradiography, the synthesis of beta-trace protein in white CNS matter could be confirmed.


[IgM antibody deficiency with normal serum IgM concentration in a family (author's transl)]. A 5 years old child with selective antibody deficiency having slight hypogammaglobulinaemia of the IgG-class, normal IgM and absence of IgA has been described. The patient had no isoagglutinins and no heteroagglutinins against rabbits. Antibodies of the IgG-class could be detected in a very low concentration. The child had repeated severe infections, f. e. 5 attacks of pneumonia. After diagnosis of selective antibody deficiency syndrome had been made, he has been treated with high dose gammaglobulin therapy every 4 weeks. The child did not have any severe infections since that time. Diagnostic and therapeutic possibilities in patients with selective antibody deficiency were discussed.


[Application of correspondance factor analysis to function testing of the thyroid]. Biological data concerning 249 thyroid patients have been treated by using correspondance factor analysis. This statistical treatment has enabled us to compare 2 types of associated tests (in vitro, in vivo) and to evaluate their respective contribution to the diagnosis of different thyroid diseases. There appeared also certain possibilities for the use of such a method in the future: determination of a biological profile for thyroid disease and for Grave's disease patients in course of treatment.


[Pollution of human milk in France by organochlorine insecticide residues]. The monthly analysis of milk mixtures from ten French human milk banks, over a period of two years, showed that there was marked pollution by organochlorine insecticide residues. The following averages were noted: hexachlorocyclohexane + hexachlorobenzene 2,75 mg/kg of pure fat, DDT and derivatives 3,24 mg/kg fat, heptachlor epoxide 0,28, dieldrin 0,23. The main insecticides were, with average values, beta hexachlorocyclohexane (1,67), hexachlorobenzene (0,98), and dichlorodiphenyldichloroethylene (2,40). All these milks were invariably very polluted throughout the year, but there were however noted some qualitative differences according to the geographic position of the milk banks: South of the Loire river, the main polluting factor was hexachlorocyclohexane, whilst North of the Seine, fairly high values were found for heptachlor epoxide and dieldrin. The special study of human milk from Lille showed a 50% reduction in pollution between 1970 and 1973. The probable routes of contamination are human food and the ill-advised domestic use of pesticides.


[Value of separation on ion exchange resins of plasma iodine fractions. Comparison with Werner's suppression test]. This was a study of 71 patients suspected of hyperthyroidism who had undergone a tri-iodothyronine suppression test. A study of circulating protein-bound iodine after separation on ion-exchange resins had been carried out before triiodothyronine administration, on the same patients. The results showed that the radioactive hormonal fraction has a better diagnostic value than radioactive serum protein-bound iodine. In 60% of cases, a study of the radioactive hormonal fraction could have permitted to avoid the use of the suppression test.


[A method for the study of relations between the lung and the reticulo-endothelial system]. The experiments reported in this paper draw attention to three new facts : 1) the reticulo-endothelial system (RES) of normal Rat can be " filled " with a non toxic, polymeric substance, PVNO, which is taken up by macrophages by pinocytosis ; 2) a broncho-pulmonary irritation leads to a discharge of PVNO by the macrophage. The macrophages are poured out into the blood stream, whence they arrive in to the lung ; 3) this discharge of a polymer as PVNO which has been previously taken in the macrophage, can happen even 10-12 months after the filling of the RES by the macromolecule.


A study of the plasma kinin-generating system in children with the minimal lesion, idiopathic nephrotic syndrome. Although the precise etiologic incitant of the minimal lesion idiopathic nephrotic syndrome of childhood is not known, it is likely that a host mechanism mediates the permeability alterations of the glomerular capillary wall resulting in massive proteinuria. As a first step in examining the possibility that local kinin release may account for the proteinuria in this disorder, two parameters of the plasma kinin-generating system, plasma prekallikrein and kallikrein inhibitor, were assayed during 27 nephrotic episodes in 21 corticosteroid-responsive children. Plasma kallikrein was assayed by means of its esterase activity on a synthetic arginine ester substrate, N-alpha-tosyl-L-arginine methyl ester (TAMe), after activation of Hageman factor by kaolin. This activity, after subtraction of spontaneous arginine esterase activity (i.e., TAMe esterase activity measured in plasma not exposed to kaolin) is derived from prekallikrein. Plasma prekallikrein activity in 11 normal children was 99.6 +/- 2.9 mumol TAMe hydrolyzed/ml plasma/hr (mean +/- SEM). Kallikrein inhibitor was quantified in arbitrary units. Kallifrein inhibitor activity in 11 normal children was 0.94 +/- 0.04 units. During the overt nephrotic syndrome, before initiation of intensive daily corticosteroid treatment, mean values were: prekallikrein, 58.5 +/- 7.24 mumol/ml/hr; and kallikrein inhibitor, 0.35 +/- 0.06 units. After corticosteroid-induced remission occurred, mean values were: plasma prekallikrein, 118.6 +/- 3.2 mumol/ml/hr; and kallikrein inhitor, 0.78 +/- 0.03 mumol/ml/hr. Both parameters were again assayed in 14 of the 21 children after complete cessation of corticosteroid treatment. Plasma prekallikrein was normal, 99.6 +/- 4.8 mumol/ml/hr; but kallikrein inhibitor was still somewhat depressed, 0.84 +/- 0.03 units. A subset of 9 patients had marked depression of plasma prekallikrein to levels less than 20 mumol/ml/hr and essentially undetectable inhibitor activity. Serum alpha-2 macroglobulin was elevated in nephrotic patients: mean value during relapse, 862 +/- 29 mg/100 ml; during corticosteroid-maintaining remission, 615 +/- 29 mg/100 ml. After cessation of corticosteroids, mean serum level was 481 +/- 20 mg/100 ml. The proportional reduction of plasma prekallikrein and kallikrein inhibitor suggested that an enzyme-inhibitor complex formed in vivo, perhaps at a local site of activation in proximity to the glomerular basement membrane. These data suggest that the plasma kinin-generating system may be the host effector mechanism subserving the increased glomerular capillary permeability in the minimal lesion nephrotic syndrome of childhood.


[Transsphenoidal hypophysectomy in metastasizing breast cancers. Experience from 160 cases]. 160 patients suffering from advanced metastatic cancer of the breast had transsphenoidal open surgical hypophysectomy. Mortality (1,8 p.cent) and morbidity were extremely low in view of the general conditions of the patients. Immediate relief of pain due to bone metastasis was obtained in 92 p.cent of cases. Such results emphasized the important point of achieving total hypophysectomy.


[Intubation of neoplastic esophageal stenoses. A new prosthesis with an antireflux device]. A preliminary communication on the excellent results of a new intraluminal prosthesis for palliative intubation of advanced oesophageal of cardial carcinoma is reported. Such prothesis includes an original and effective device to avoid gastro-oesophageal reflux and the ensuing peptic oesophagitis. Piloroplasty is considered convenient as an adjuvant in the prevention of gastro-oesophageal reflux.


Immunochemical evidence for a common variable region in three immunoglobulin classes in the same individual. One IgG1(kappa), one IgM(kappa), and one IgA1(kappa) monoclonal (M)-component were purified from one human serum. Rabbit antisera were raised against the IgG and IgM M-components and were absorbed until specific for idiotypic determinants on these molecules. All three M-components gave reactions of immunological identity when tested by double radial immunodiffusion with either of the two idiotype-specific antisera. Both heavy and light chains were isolated from each of the three M-components and all preparations inhibited formation of idiotypic precipitates. None of these preparations formed precipitates with idiotype-specific antisera alone. When heavy or light chains of one M-component were hybridized with light or heavy chains from the other M-components the resultant molecules precipitated with anti-idiotypic serum. Hybrids with chains from polyclonal IgG were not precipitable with such antiserum. These results indicate that the variable region of the heavy chains of these M-components of three different immunoglobulin classes are closely similar, if not identical.


Purification of RNA-instructed DNA polymerase from human leukemic spleens. Particles possessing a density of 1.16 g/ml and encapsulating a 70S RNA and a RNA-instructed DNA polymerase (reverse transcriptase) have been prepared from the spleen of a patient with chronic lymphocytic leukemia. These particles have been converted to cores with a density of 1.26 g/ml and containing the enzyme-RNA complex, in complete analogy to the known RNA tumor viruses of avian and murine origin. The reverse transcriptase was purified from the cores by column chromatography to a stage showing a single major protein band of 70,000 daltons in a gel electrophoresis. The enzyme was capable of transcribing heteropolymeric RNA into DNA complements as demonstrated by specific back hybridization to template RNA. The leukemic spleen would appear to represent an important source of this enzyme, as well as other potentially important leukemia-specific reagents.


Filamin, a new high-molecular-weight protein found in smooth muscle and non-muscle cells. A new high-molecular-weight protein, named filamin, was isolated from chicken gizzard. In chicken gizzard, filamin is present in an amount approximately 30-40% of that of myosin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of highly purified filamin revealed a single polypeptide of about 250,000 daltons. Rabbit antibody directed against purified chicken gizzard filamin did not crossreact with myosin purified from the same source. By the use of microcomplement fixation and indirect immunofluorescent staining with antibodies to chicken gizzard filamin, an antigenically similar or identical protein was found to be widely distributed both in other organs of the chicken and in cultured cells of other species, but not in chicken skeletal muscle. In cultured cells, filamin was found largely to be arranged as a filamentous array very similar to that found for myosin. These data imply that filamin is a widely occurring and chemically conserved component of filaments is smooth muscle and non-muscle cells.


Immunochemical characterization of murine H-2 controlled Ss (serum substance) protein through identification of its human homologue as the fourth component of complement. The S region of the mouse H-2 complex is genetically defined by a series of alleles, or pseudoalleles, which control the level of a serum globulin, Ss, and of its allotypic vairatn, Slp. In contrast with the products of other genes in the complex, no homologue of the Ss protein has been found in other species, except the rat. In the present study, a component in human plasma was identified which cross-reacts with anti-mouse Ss, and which also displays electrophoretic and size similarities to the mouse Ss protein. This component was isolated and characterized immunochemically as the fourth component of human complement (C'4 or beta1E globulin). Several antisera monospecific for human C'4 cross-react with the mouse Ss protein and detect its genetically determined quantitative variations.


Induction of an immune response through interaction of helper cells with an immune complex bound to the surface of B cells. Adoptive anti-trinitrophenyl (Tnp) responses were elicited from Tnp-hemocyanin (Tnp-KLH)-primed cells by challenge with an immune complex of KLH and a Tnp conjugate of the Fab fragment of rabbit anti-KLH. Removal of T cells by treatment with anti-Thy 1.2 (phi C3H) + complement abolished this effect. Tnp-KLH primed cells that had been incubated with a Tnp conjugate of pneumococcal polysaccharide type III (Tnp-SIII), and which were then unable to respond to Tnp-KLH, made an anti-Tnp response upon incubation with rabbit anti-Dnp when transferred together with cells primed with rabbit gamma globulin. Since Tnp-KLH was not added in these experiments, it would appear that the cells were triggered by immune complexes on their surfaces consisting of Tnp-SIII and rabbit anti-Dnp with the help of T cells primed with rabbit gamma globulin; the presence of free antigen was evidently not required. Therefore, antigen-antibody complexes, when bound to the surface of B cells, may mediate T-B cell cooperation in the humoral immune response.


Structural features of double-stranded polyribonucleotides required for immunological specificity and interferon induction. Purified antibody to poly(adenylic acid)-poly(uridylic acid) was used in quantitative microcomplement fixation assays to detect conformational variations among several double-helical polyribonucleotide analogs of poly(adenylic acid)-poly(uridylic acid) or poly(inosinic acid)-poly(cytidylic acid) that had been previously evaluated for their ability to induce interferon. Modification at the furanose 2'-position of one or both strands resulted in a dramatic decrease in serological reactivity. Most modifications of the bases caused smaller serological changes, and no base modification caused complete loss of reactivity. The reaction patterns support the conclusion that the structure of the furanose and the overall conformation of the helix are critical in the formation of antigenic determinants. The backbones of both strands appear to be involved in forming a single antigenic site, and base modifications may alter the steric relationship between the backbones. In addition, the same structural changes that substantially alter recognition by antibody also lead to large changes in the interferon-inducing ability of the nucleic acid.


Activation of T and B lymphocytes in vitro: presence of beta2-microblobulin determinants on allogeneic effect factor. The biologically acitve entity termed allogeneic effect factor, which is produced by alloantigen-activated T cells and has been shown to regulate triggering and differentiation of B lymphocytes in vitro, has previously been demonstrated to possess antigenic determinants coded by genes in the I region of the H-2 gene complex of the mouse. The studies presented here provide evidence that allogeneic effect factor also possesses antigenic determinants identical or cross-reactive with beta2-microglobulin. These observations suggest an important role for beta2-microglobulin in the mechanism of cell-cell interactions and the consequences of such interactions on lymphocyte triggering and differentiation.


Stimulation and inhibition of anti-hapten responses in guinea pigs immunized with hybrid liposomes. Guinea pigs were immunized with liposomal model membranes containing phosphatidylethanolamine (PE) or glycerophosphorylethanolamine (GPE) derivatives in which the amino function was substituted with either dinitrophenylaminocaproyl (Dnp-Cap) or mono(p-azobenzenearsonic acid)tyrosyl (ABA-Tyr) residues. Previous studies have demonstrated that hapten-specific antibodies are elicited by DNP-Cap-PE or ABA-Tyr-PE sensitized liposomes and that cell-mediated immunity is induced by ABA-Tyr-PE (but not Dnp-Cap-PE) sensitized liposomes. These liposomes differ from conventional immunogens in which haptens are covalently attached to immunogenic carriers. This investigation describes two new aspects of liposomal immunogenicity in animals immunized with hybrid liposomes containing both Dnp-Cap-PE and ABA-Tyr-PE. (1) Stimulation of the anti-Dnp response by incorporation of increasing amounts of ABA-Tyr-PE; (2) inhibition of anti-ABA antibody formation by incorporation of increasing amounts of DNnp-Cap-PE. The two phenomena are dependent on the presence of each determinant in the same lipid bilayer. Thus, entrapment of the water-soluble deacylated derivative of ABA-Tyr-PE (i.e., ABA-Tyr-GPE) in a aqueous compartments of Dnp-Cap-PE sensitized liposomes does not enhance anti-Dnp antibody production. Similarly, entrapment of the non-amphipathic derivative of DNP-Cap-PE (i.e., Dnp-Cap-GPE) within ABA-Tyr-PE sensitized liposomes does not suppress anti-ABA antibody formation. Furthermore, mixtures of Dnp-Cap-PE sensitized liposomes and ABA-Tyr-PE sensitized liposomes neither stimulated nor inhibited the anti-hapten responses. These results indicate that preparation of hybrid liposomes with different N-substituted PE derivatives provides an extremely convenient method for controlling hapten and/or immunologic carrier determinant density.


Multiple active sites on human interferons. Human interferons stimulated in peripheral leukocytes and foreskin fibroblasts are active in cultures of human and rabbit cells. The dominant factors in leukocyte and fibroblast interferons responsible for antiviral activity in rabbit cells were shown to be antigenically distinct from each other as well as from rabbit interferon. In addition, leukocyte interferon contained also a minor component with antigenic determinants characteristic of fibroblast specificity, which could be isolated by affinity chromatography on Sepharose-bound antibodies directed against firboblast interferon. Neutralization tests with selected anti-interferon sera suggested that the antiviral activities of leukocyte and fibroblast interferons in human and rabbit cells were associated with single molecules. A model is proposed where molecules of human interferon contain multiple reactive sites each of which is capable of interaction with cells of a different species. The number and distribution of these determinant sites may vary with the source of the human interferon and account for the differential in antiviral protection expressed in homologous and phylogenetically unrelated host cells.


Oncornavirus-like particles released from arginine-deprived human lymphoblastoid cell lines. Type-C RNA tumor virus particles were released from three different human lymphoblastoid cell lines after incubation in arginine-deficient medium. The released virus-like particles were characterized by (a) their ability to band in sucrose gradients at a density of 1.16-1.18 g/ml; (b) the presence of an RNA-directed DNA polymerase activity resembling that of the oncornaviruses; and (c) isolation of cores that band at a density of 1.26-1.27 g/ml in sucrose gradients. Examination of the arginine-deprived human lymphoblastoid cell line strain P3HR-1 by electron microscopy revealed the presence of C-type particles in the intracellular spaces.


Carcinoembryonic antigen: evidence for multiple antigenic determinants and isoantigens. Carcinoembryonic antigen (CEA) preparations, from various sources were compared by radioimmunoassay. The preparations studied included four CEA standards (CEA-Roch, CEA-Montreal, CEA-City of Hope, and CEA-British) and CEA from serum and liver metastases of a patient with cancer of the colon who had an extremely high concentration of serum CEA (more than 26,000 ng/ml). The data indicate that the CEA-Roche standard differs significantly from the other three CEA standards tested, and that the serum CEA from the patient was antigenically different from currently available CEA standards as well as from the CEA obtained from the patient's own liver metastases. These antigenic differences were reflected in radioimmunoassay inhibition curves that were different and that were not affected by perchloric acid extraction of CEA. Because of the antigenic variation in the serum CEA, markedly different CEA concentrations (varying by three orders of magnitude) were measurable by two different antisera (Roche and Montreal). All the various CEA standards and samples cochromatographed on columns of Sepharose-6B, despite the large antigenic variation. We postulate that CEA consists of a family of "isoantigens" with multiple antigenic determinants. We identified a serum CEA isoantigen that was different from the currently available standards. Consequently, we believe that results of radioimmunoassays currently used for CEA measurement may not represent absolute concentrations of serum "CEA", but may reflect the binding affinity of different isoantigens to a particular polyvalent CEA antiserum.


Endogenous immune complex nephropathy associated with malignancy I. Studies on the nature and immunopathogenic significance of glomerular bound antigen and antibody, isolation and characterization of tumor specific antigen and antibody and circulating immune complexes. Three patients with clear cell renal carcinoma and one with another intrarenal malignancy were studied for the presence of glomerular localized immunoglobulins, complement components and tumor specific antigen and antibody by immunofluorescence. To determine the association and elucidate the pathogenic mechanisms involved in the relationship between tumors and glomerular deposits, antibody eluted from tumor tissue and renal glomeruli, cryoproteins, serum antibodies and rabbit antisera to tumor tissue were tested for specificity to antigen. The relationship between tumor antigens and the lipoprotein antigen localized in normal proximal tubular brush border (RTE) and the small bowel mucosa, was studied by immunofluorescence, absorption and blocking studies as well as complement fixation. Immunoglobulins and complement components were localized in the glomeruli and tumor membrane of all patients. Sera and glomerular fixed antibody from three patients with renal cell carcinoma localized to normal proximal tubular brush border and jejunal mucosa as well as to tumor membrane and the glomeruli and proximal tubules of all of these three patients. Anti RTE activity was also detected by complement fixation. Immunologic similarity between RTE and renal cell carcinoma antigen was confirmed by absorption studies. Furthermore, cryoprecipitable complexes of tumor antigen and specific antibody were isolated from the serum. The tumor antibody was immunologically similar to RTE. In the other case the rabbit anti-tumor antibody and the patient's serum fixed to the tumor membrane and kidney of the patient but did not show cross reactivity with the renal cell carcinoma or RTE. These studies suggest that the tumor antigen in renal cell carcinoma is similar to RTE and the glomerular deposits represent tumor antigen and antibody complexes. In addition these investigations support the hypothesis that tumor immune complexes are significant in the glomerular lesions, and that the origin of renal cell carcinoma is in the proximal tubule. The investigations also show that tumor antibodies are specific for tumors of the same morphological type but not for other tumors in the same tissue. Moreover, the renal glomerulus appears to be a chosen anatomic site for deposition of tumor antigens and antibodies and studies of the kidney may provide insight into the nature of tumor antigens and antibodies. Cryoprecipitation appears to be a valuable method in isolation of tumor complexes and characterization of tumor specific antigen and antibody.


Proton-induced x-ray emission analysis--a promising technique for studying the metal content of plants and soils. The ease of employing proton-induced X-ray emission analysis (PIXEA) to studies relating metal content of soils to metal uptake in plants was aptly demonstrated in an investigation concerning the effect of automotive pollution on the abundance of about 16 elements accumulated in ribwort plantain and its surrounding soil. Elemental concentrations were shown to be dependent on the age of the plant leaves, as well as the distance from the roadside.


The history and progress of serologic immunotherapy and radiodiagnosis. The early demonstration of immunologic specificity of antibodies and the discovery of tumor antigenic specificity are reviewed in the light of experimental and clinical attempts to use such reagents in the management of cancer. Recent results in regard to tumor antigens and radiolabeled antibody preparations are shown to be practical for experimental diagnosis and therapy and potentially for similar clinical purposes.


Chemical nature of atrial specific granules. The specific granules are argentafugic when ultrathin sections of Araldite-embedded atria are stained according to the periodic acid-thiocarbohydrazide-silver proteinate technique of Thiery. The entire core of the atrial specific granules is moderately positive after ultrathin sections of glutaraldehyde-fixed, glycol methacrylate- (GMA-) embedded atria are stained with phosphotungstic acid at a low pH. A similar reaction is shown by the cell coat, intercalated discs, residual bodies (C-granules), and Z-discs, as well as by a very small portion of the Golgi complex. Analogous results are obtained with semithin sections of GMA- embedded atria stained according to the periodic acid-Schiff (PAS) technique. In ultrathin sections of GMA-embedded atria stained with dialyzed colloidal iron (DI), the cell coat of the cardiocytes is positive, unlike all the other cytoplasmic organelles. When ultrathin sections of GMA-embedded atria are incubated with proteolytic enzymes (pronase, pepsin, or trypsin), atrial specific granules and Z-bands and, to a much lesser degree, cell coat and sarcolemma are selectively digested. Proteins are also distinctly demonstrated in the paranuclear specific granules by a variety of histochemical techniques. These results indicate that atrial specific granules are rich in proteins and possess a weak complement of complex carbohydrates.


Passage of immune complexes through vascular walls. Evidence for the role of an immediate hypersensitivity mechanism and its mediator: platelet-activating factor. An anaphylactic mechanism involving IgE-sensitized basophils is necessary for immune complex deposition in rabbits. Basophils under stimulation by the IgE-bound antigen actively release a platelet-activating factor. Increased vascular permeability results from action of vasoactive amines from basophils and from aggregated platelets and possibly direct action of PAF on vascular walls. This allows trapping of circulating complexes along vascular membranes. Once deposited complexes produce tissue injury by neutrophil-complement-dependent mechanisms. However, complex deposition seems pathogenic in itself for the glomerular basement membrane. PAF is a 1,100-dalton MW lipid, with hydrophobic properties and positive charge; it exists in human basophils. Patients suffering from SLE or undergoing anaphylactic shocks exhibit marked reduction of basophil counts and PAF level. We propose that anaphylactic increase of vasopermeability may be a common feature at early stages of many immune diseases associated with severe structural injury.


A direct spectrometric method for determination of the concentration of available bilirubin binding sites in serum using bromphenol blue. A simple and rapid direct spectrometric micromethod for the determination of the concentration of available bilirubin binding sites of serum is given. The method measures - by means of difference spectrometry - the maximal change in absorbance when serum is added to a buffered solution of bromphenol blue (BPB). This dye fulfills a set of requirements given and examined. The binding properties of bilirubin, BPB, and some related dyes are spectrometrically investigated, and differences between the properties of human and bovine albumin and between such albumin preparations and human serum are shown and discussed.


Renal handling of beta2-microglobulin in experimental renal disease. Renal extraction and urinary excretion of 125I-labelled beta2-microglobulin was studied in rats. The effect of ischaemic renal injury, experimental pyelonephritis, and unilateral nephrectomy was investigated. The tubular secretion of o-iodohippurate (OIH) was measured for comparison. The urinary excretion was calculated as the ratio between the clearance of protein and the glomerular filtration rate. The glomerular filtration rate was estimated as clearance of polyethylene glycol (PEG 1000). The renal arteriovenous concentration difference was lower for beta2-microglobulin than for PEG 1000 IN ALL THE EXperimental groups. In unilateral renal disease the beta2-microglobulin excretion of the intact kidneys was similar to that of the diseased kidneys. A significant differences was noted only after ischaemic renal injury. The same was found for OIH. After removal of the intact kidneys the excretion of beta2-microglobulin increased about 10-fold in pyelonephritic animals and 2- to 30-fold in animals with ischaemic renal injury. One hour after unilateral nephrectomy in normal animals the ratio increased about 50 per cent. The tubular secretion of OIH did not change noticeably. It is concluded that the glomerular filtration is a main step in the intrarenal catabolism of beta2-microglobin and that its urinary excretion is considerably influenced by a reduction in the functioning kidney mass.


Long-term prognosis of 66 permanent anterior teeth with root fracture. A longitudinal clinical and radiographic follow-up study was made of all permanent teeth with root fractures referred to the Oslo University Department of Pedodontics between 1953  and 1972 (n = 66). The material included 51 patients aged 6-21 years. The mean observation period was 5.2 years, ranging from 1 to 19 years. The present report documents background data and the long-term results. Two teeth with exarticulation of the coronal fragments (3%) were immediately extracted. Repair of the fracture area occurred in 51 teeth (77%). Pulp necrosis was found in 13 teeth (20%), nine of which were successfully treated endodontically; only four teeth had to be extracted. Several factors were found to influence the prognosis, most notably the degree of dislocation of the coronal fragment. The localization of the fracture influenced repair only slightly. Despite somewhat increased mobility in some cases, the longevity of teeth with fractures even in the coronal third of the root was not significantly shortened. It is concluded that when optimally treated by repositioning, fixation and relief of occlusion, anterior teeth with root fracture have a favorable prognosis. Even when pulp necrosis occurs, the long-term prognosis is good.


Contamination of anti-immunoglobulin reagents with antibodies to beta 2-microglobulin and other unrelated antigens. Effects on immunofluorescence staining of human lymphocytes. IgG fractions from three of four rabbit antisera to Bence Jones proteins of chi-type were found to contain antibodies to beta 2-microglobulin and to stain 80%-100% of human blood lymphocytes by indirect immunofluorescence. Antibody fractions from these sera, which contained anti-beta 2-microglobulin but not anti-Ig, stained all lymphocytes, whereas the isolated anti-Ig antibodies (anti-chi) stained only a minor cell population. In both instances, the specificity of the staining was confirmed by absorption experiments. One antiserum to the constant half of lambda-type Bence Jones protein also contained antibiodies to beta 2-microglobulin and stained all lymphocytes. Four other anti-lambda reagents contained no antibodies to beta 2-microglobulin and stained at most about half of the lymphocytes. The antigen responsible for this staining is unknown. The isolated anti-immunoglobulin antibodies (anti-lambda) stained only 5%-10% of the lymphocytes. Antisera to serum IgG or its fragments were free of antibodies to beta 2-microglobulin and stained only 10%-25% of the lymphocytes. This staining was in all instances due to antibodies to human immunoglobulin. Five of eight undiluted sera from normal rabbits with no detectable antibodies to human immunoglobulin or beta 2-microglobulin stained 25%-60% of the lymphocytes. This staining rapidly disappeared on dilution.


Immunological characterization of human glomerular basement membrane antigens. Normal human glomerular basement membrane (H-GBM) was solubilized by collagenase and subjected to crossed immunoelectrophoresis with rabbit antibodies against H-GBM. Seven precipitates appeared with the mobility of alpha, beta, and gamma globulins. Only two of these precipitates might be specific for GBM, since the other precipitates disappeared after absorption of the antiserum with liver and placenta. In normal human urine one precipitate, cross-reacting with one of the H-GBM precipitates, was found; this precipitate could also be demonstrated in human placenta and liver.


Survey of the sputum cytology service in England and Wales. Of the 231 laboratories carrying out sputum cytology in England and Wales, information on the staffing, laboratory procedures, workload, records, and results for the year 1971 was obtained from 228 (98.7%) by means of a questionnaire, which was completed in 1973 in 81% of the laboratories and at the end of 1972 in the remainder. At the time the questionnaire was completed, the number of laboratories per region offering a service ranged from 6 to 25. Most of the processing of specimens was done by technicians and non-medical screeners (technicians without recognized qualifications) and most of the microscopy by pathologists and technicians. Papanicolaou and/or haematoxylin and eosin stains were used in 97% of the laboratories. During 1971, the number of specimens tested per laboratory ranged from 9 to 6000. A median of 381 specimens and 165 patients were tested per laboratory. Almost all specimens were submitted by hospitals and chest clinics and only 1% by general practitioners. An estimated median of 6.5% of all cytological specimens were sputum, most of the remainder being cervical smears. Laboratories varied greatly in their positive (0%-24%) and doubtful positive (0%-27%) rates. An estimated 63 000 patients had their sputum tested in 1971, and about 6300 had positive reports, rather less than one-quarter of all patients diagnosed as having lung cancer in England and Wales in 1971. The cost of the service was estimated to be of the order of pound375 000 at 1974 prices; that is, pound2.50 per specimen or pound6.00 per patient tested. An improved service might best be provided by district general hospital laboratories testing at least 1000 specimens per year so that a high level of accuracy in reporting could be assurred. A large proportion of the microscopy might be done by non-medical graduates and technicians under the supervision of pathologists.


Optic nerve axoplasm and papilledema.

Optic nerve axoplasm and papilledema. A detailed review of optic nerve axoplasm is presented. A number of hypotheses have been postulated for the pathogenesis of papilledema associated with increased intracranial pressure. These hypotheses, mechanical and nonmechanical, are critically evaluated in relation to five essential features of papilledema. Theories, as well as clinical and experimental studies, of axonal transport are reviewed, and a new hypothesis is proposed: Papilledema is primarily a mechanical, nonvascular phenomenon in which an excess amount of extracellular fluid is present in the prelaminar region of the optic disc and the accumulation of that fluid results from the leakage of axoplasm from optic nerve fibers which are compressed posterior to the lamina cribrosa of the optic disc. The authors believe that this is the only existing hypothesis consistent with all the known facts about papilledema. Discussions by Drs. J. Terry Ernest, Thomas R. Hedges, and S. S. Hayreh follow the review.


The fibrinolytic system and haemostasis. The fibrinolytic enzyme system appears to be a parallel development with the plasma coagulation pathways involved in haemostasis. Both systems have common activation mechanisms, inhibitors and the enzymes involved have considerable homology. In addition, blood cells show capabilities for involvement in each of these systems. There is disagreement over the possible contribution of the fibrinolytic mechanism to normal fibrinogen catabolism either by the lysis of continually forming fibrin, or by degradation of circulating fibrinogen. The occurrence of systemic fibrinolysis is associated with haemorrhagic tendency which may be due to a number of factors including degradation of coagulation factors, the anticoagulant effect of F.D.P. and the lysis of alpha-chain crosslinking sites. A clinical study of patients treated with streptokinase indicates that a fall in plasma fibrinogen levels and increase in serum F.D.P. levels are correlated with the haemostatic defect.


Blood changes in atherosclerosis and long after myocardial infarction and venous thrombosis. The following clinical groups of volunteers were studied: patients long after recovery from myocardial infarction (MI), others after recovery from deep vein thrombosis (DVT), patients with intermittent claudication, with diabetes, and male and female controls who were well matched. All were subjected to many platelet and clotting tests together with clinical, biochemical and haematological measurements in an attempt to find long term abnormalities in these various diseases. The male MIs differed very significantly from the controls in having much more heparin neutralizing activity (P less than 0.001)and less anti-thrombin (P less than 0.01). Less significantly, some bleeding time tests indicated less bleeding and the patients' platelets were larger. The females with MI had in general the same abnormalities but to a lesser degree. The patients with intermittent claudication, none of whom had a history of MI, had almost the same abnormalities and to the same degree. In deep vein thrombosis the heparin neutralizing activity was also clearly increased; the other tests were generally in the same direction but many were not significant. The diabetics had shorter bleeding times but little else abnormal relative to the controls, suggesting a different pathological process. When all male patients and controls were "scored" according to the degree of atherosclerosis there was a close overall correlation between the degree of atherosclerosis and the increase in the HNA level (r = --0.50, n = 66, P less than 0.001) and the decreased anti-thrombin (r = 0.25, n = 66, P less than 0.05).


Local treatment of recent arterial thromboembolic occlusions with brinase. Brinase was administered intra-arterially in 16 patients with thrombotic or embolic arterial occlusions. Angiography could be performed before and after treatment in 13 patients. Thrombolysis was obtained in 3 of 9 patients with thrombotic and in 3 of 4 patients with embolic occlusions. In 3 patients severe local side effects occurred.


[Letal rate and late complications following palliative surgery of transposition of the great arteria (author's transl)]. Between 1966 and 1973 145 palliative procedures were carried out on 110 infants with transposition of the great arteries. There were 23 early deaths (20.9 per cent). 85 per cent of the survivors were controlled till the end of 1974. Five late deaths and four cases of peripheral artery embolism were regarded. These complications, however, took place several years after palliation. The development of pulmonary hypertension doesn't seem to be as high as it is suspected sometimes. These results justify our conception, not to perform the Mustard-operation during the first two years of life.


Identification of tissue-specific antigens and concanavalin A receptors on rat epidermal cells using a radio-immunoprecipitation method. Iodination with lactoperoxidase - 125I- - H2O2 was used to label surface components of rat epidermal cells. Lysis of the cells in non-idet P40 resulted in the solubilization of tissue-specific antigens and of concanavalin A receptors. These specificities were demonstrated using a radio-immunoprecipitation method. The tissue-specific antigens were recognized using absorbed rabbit anti-rat epidermal cell sera (Lloyd & Darnule 1974); they reacted with two low molecular weight components in the lysate (9,000 and 12,000 daltons). Concanavalin A reacted with three major components. Two had high molecular weights (75,000 and 95,000 daltons). The possibility that one of these components was radioiodinated lactooperoxidase, which would have reacted with concanavalin A, was disproved. Another component (which occasionally appeared as two peaks) was similar in size to the species detected by the tissue-specific antisera. Their non-identity was, however, demonstrated by the finding that the two specificities could be precipitated independently of each other.


LD antigens associated with HL-A8 and myasthenia gravis. The finding that the SD antigen HL-A8 is associated with myasthenia gravis has raised the question as to whether or not there is also an association between some specific LD antigen and myasthenia gravis and/or HL-A8. MLC technique was used to study LD antigens in 33 myasthenic patients. The cells of a myasthenic patient who was homozygous for both HL-A and LD products were used in MLC tests as stimulators with HL-A8 bearing cells from 24 myasthenics, their 17 relatives and 16 controls and to non-HL-A8 cells from nine myasthenics and 16 controls. At least three different LD genes were found to be associated with HL-A8. One of them, called LDm' was present in 17 (63 %) of the 27 HL-A8-bearing haplotypes of myasthenics, and in nine (47 %) of the 19 HL-A8-bearing haplotypes of controls. In our study LDm was not found in the 84 haplotypes devoid of HL-A8. LDm is strongly associated with HL-A8 and through this also with myasthenia. Calculated from phenotype frequencies for HL-A8 and its association to LDm' the LDm frequencies are 9 % in the control population, 30 % in myasthenics and 48 % in young females with onset of myasthenia below 35 years. LDm had no indendent correlation with any of the clinical parameters of myasthenia gravis, even though is was found more often in HL-A8+ females with the onset of myasthenia before 35 years than in the whole myasthenic group. LDm appears to be similar to LD8a antigen.


Lymphocyte activating alloantigens on human epidermal cells. Human epidermal cells dissociated by repeated trypsinization of skin explants were stimulatory to allogeneic lymphocytes in mixed lymphocyte skin-cell cultures. After elimination of the lymphocytes proliferating in response to a particular allogeneic lymphocyte donor by a "hot" pulse of 3H-thymidine of high specific activity, the viable lymphocytes remaining in culture were still capable of responding to stimulation by epidermal cells from the same donor. The response towards allogeneic epidermal cells could only partially be eliminated by 3H-thymidine treatment. This treatment, however, also partially eliminated the secondary stimulation response towards lymphocytes from the epidermal cell donor used initially, but did not alter the response to lymphoid or epidermal cells from an unrelated third party donor. In addition, HL-A antisera, which specifically inhibited the stimulatory capacity of mitomycin-treated lymphocytes that carried the relevant HL-A antigen also inhibited the stimulatory properties of epidermal cells from the same donor.


Rhesus lymphocyte alloantigens. III. Identification of new antigens. Rhesus monkey lymphocytotoxic alloantisera were tested in 437 random unrelated monkeys and in members of 19 pedigreed families. Groups of sera or individual sera identified 11 specificities. Genetic analysis of the associations of these antigens revealed behavior consistent with the current concept of the major histocompatibility complex of several mammalian species including man in that the antigens could be grouped into two series of closely linked co-dominantly expressed alleles.


Cell mediated lympholysis in man. Patterns of killing power in relation to the HL-A system. In general it can be said that the HL-A (LA and FOUR) antigens seem to be of major qualitative and partly quantitative importance for the degree of destruction in Cell Mediated Lympholysis (CML). This has been established by analyses of 1299 effector/target combinations involving 97 different healthy individuals. The main observation is that the cytolytic capacity of MLC effectors is positively ranked with the number of HL-A (LA and FOUR) antigens introduced by the incompatibility between stimulator, effector and target lymphocytes. However, the fact that not all cytolytic situations can be explained by the HL-A system stresses that other specific antigens (Target Determinants) may interfere with the Cell Mediated Lympholysis.


Cell mediated lympholysis in man. Varying strength of the HL-A (LA and FOUR) antigens as sensitizing or target determinants. The gene products of the LA and FOUR loci of the human Major Histocompatibility Complex (MHC) are generally considered to be a major target in the Indirect Cell Mediated Lympholysis (ICML) test. Within most experiments, a positive correlation exists between the number of HL-A antigens challenged and lympholysis. When different experiments are collated this correlation is less obvious. This discrepancy might be caused by differences between the individual HL-A antigens involved in the afferent phase (MLC) and/or the efferent phase (CML). In 28 experiments involving 97 unrelated individuals we have compared statistically 12 different antigens governed by the LA and FOUR loci. When only one of these antigens is challenged in ICML, target lymphocytes are lysed to different degrees allowing a significant classification of the antigens into different groups, which do not coincide with the classification in the LA and FOUR series antigens. It is concluded that the antigens of the HL-A system are not of equal importance when challenged separately in ICML. The existence of a separate CML locus and a corresponding linkage disequilibrium to the SD loci of the MHC region is suggested.


Cell mediated lympholysis in man. The HL-A third locus antigen, AJ (W20) as sensitizing and/or target determinant. The Indirect Cell Mediated Lympholysis (ICML) test has become a reliable in vitro tool in investigations of the immunogenetic background of the cellular immune response. It is well established that the antigens of the LA and FOUR loci of the human Major Histocompatibility Complex (MHC) or antigens governed by closely linked loci exhibiting a marked linkage disequilibrium are of major qualitative and quantitative importance in ICML. This communication is concerned with the importance of the HL-A third series antigen AJ (W20) on lympholysis in ICML. From investigations of 47 combinations where only the AJ (W20) antigen may be attacked, it is concluded that this antigen is, in itself, a poor ICML target determinant, but that AJ (W20) may function as a stronger ICML target determinant in concert with a FOUR antigen, although the actual FOUR antigen cannot be attacked.


Studies on the major histocompatibility system in patients with ankylosing spondylitis. One hundred % of 66 patients with pelvospondylitis were found to have HL-A27, whereas the frequency of the antigen in 60 patients with sacroiliitis was 70 %. The difference in frequency of HL-A27 between the two patient groups is statistically highly significant. This implies that tissue typing could be of clinical importance for a prognostic evaluation of patients with sacroiliitis. Studies in families with several HL-A/MLC identical sibs indicated a relatively low penetrance of disease in those carrying the gene for increased susceptibility to development of AS. No association of AS to particular MLC alleles was found either in families or in several SD-2 identical AS patients.


Steroid receptors in the human prostate. 1. Estradiol - 17beta binding in benign prostatic hypertrophy. Agar gel electrophoresis and ultracentrifugation on continuous sucrose gradients revealed the presence of a 4S estradiol 'receptor' in cytosols of samples of human benign hyperplastic prostate tissue. High affinity (charcoal stability) and saturability (disappearance with excess competitor) binding characteristics of the molecular species concerned facilitated its clear distinction from similarly migrating serum albumin-steroid complexes. Our data, obtained with human serum, purified human albumin and albumin-enriched cytosol strongly suggest that agar gel electrophoresis, when used alone, may lack specificity for the quantification of estrogen 'receptors'. Radioligand binding to these molecules may be obscured by similarly migrating albumin-steroid complexes which fail to dissociate during electrophoresis. We advocate the inclusion of competitor experiments to improve the specificity of the method.


[A contribution to the information and instruction of patients in periodontological practice]. Starting from the necessity of expanding the periodontal care of the population, the author points to the special importance of the patient's education, information and instruction in the periodontal practice and to the employment of dental nurses specialized in dental and oral hygiene. The author's approach which is based on an audiovisual method is explained; its advantages and disadvantages are discussed.


Efficacy of leucocyte-poor red blood cell suspensions prepared by sedimentation in hydroxyethyl starch. Hydroxyethyl starch (HES), which is licensed for human use as a plasma volume expander, has red blood cell sedimenting properties similar to high molecular weight dextran (HMWD). The present studies document the effectiveness of commercially available HES in preparing leukocyte-poor red blood cell suspensions (L-PRBCs) from donor whole blood and packed red blood cells stored in CPD up to 120 hours prior to processing. The effect of HES sedimentation on final white blood cell count, on erythrocyte p50 and 2,3-DPG concentration, and on short- and long-term in vivo survival of 72-hour stored donor red cells are described, along with favorable clinical experience with over 300 transfusions of HES-sedimented L-PRBCs. Since the method requires no special equipment, it is immediately available for widespread use.


Hydroxyethyl starch as an experimental adjunct to leukocyte separation by centrifugal means: review of safety and efficacy. A total of 1,921 leukapheresis procedures have been performed on 532 normal and CML donors at six research institutions, for the purpose of supporting granulocytopenic leukemia patients during infectious episodes. The addition of HES alone or in combination with either etiocholanolone or dexamethasone, resulted in a significant increase in the numbers of leukocytes (granulocytes) harvested by continuous and noncontinuous flow centrifugation. Normal donors participating in these programs were monitored prior to and immediately following each procedure by standard laboratory methods which revealed no serious or abnormal changes occurring as a result of the procedure in those undergoing single or multiple donations with these agents. CML donors tolerated the addition of only HES well, as evidenced by the lack of toxic reactions in three donors undergoing 101 to 121 procedures.


Harvesting of granulocytes using a hydroxyethyl starch solution. The need for sophisticated ocmponent therapy has resulted in improved techniques for obtaining concentrates of platelets and granulocytes. The use of single donors as a source for these products is advisable to avoid multiple sensitizations. Obtaining concentrated granulocytes represents a problem because of the difficulty in separating granulocytes from red blood cells by differential centrifugation or sedimentation since the specific gravities are similar. Hydroxyethyl starch (HES) makes the separation more effective. A solution made of 250 ml of 6 per cent HES, 250 ml of distilled water, and 15 g of sodium citrate in 30 ml distilled water provided a satisfactory anticoagulant solution for this purpose. The granulocytes collected averaged 49 per cent of the total available in the processed blood; the platelets averages 82 per cent. A satisfactory yield could thus be obtained from a single donor, and this could be repeated several times in a month. The value of ABO, Rh, and HL-A compatibility between donor and recipient probably increases the viability and safety of this procedure. The Haemonetic No. 30 Cell Separator provided an easy and rapid method for this procedure.


Short-term and long-term effects of plasmapheresis on serum proteins and immunoglobulins. In order to evaluate the short-term and long-term effects of plasmaapheresis on serum proteins and immunoglobulins, the levels of alpha1, alpha2, beta, and gamma globulins, and IgG, IgA, and IgM were measured and statistically evaluated in 41 active plasmapheresis donors donating 500 to 1,000 ml of plasma weekly for up to three years. During the initial four months of plasmapheresis, the percentage of alpha1 and alpha2 globulins manifested a statistically significant rise and the IgG, IgA, and IgM concentrations declined. By the end of ten months, only the IgM continued to be depressed. Although the concentration of IgM continued to show a statistically significant decline for three years, it remained well within the normal range of values for our laboratory. Although no statistically significant difference existed between the baseline value of albumin and the level reached at the end of the third year, a gradual rise was followed by a decline in this interval. Most of the statistically significant alterations of serum protein and immunoglobulins occurring in plasmapheresis donors are seen in the initial six months of plasmapheresis. A falling serum protein in this time period is most likely an indication of declining immunoglobulins. It is feasible and appropriate to measure the donor's total serum protein at the time of each plasmapheresis. Any untoward reduction in this value necessitates quantification of the serum immunoglobulins. Routine measurement of the immunoglobulins in the face of normal total serum protein can be performed on a less frequent basis as is presently recommended by accrediting agencies, although this study should be performed more often during the first six months of a serial plasmapheresis program.


The effect of hydroxyethyl starch on in vitro platelet and granulocyte function. The effect of hydroxyethyl starch (HES) on granulocyte and platelet functions was assessed. No alteration of granulocyte viability, morphology, phagocytic ability, or bactericidal capacity was detected after incubation with 6 per cent HES at 25 C for two hours. Platelet morphology, size distribution, aggregation, nucleotide and serotonin release, and platelet factor-3 availability were also unchanged after exposure to HES. It is concluded that HES has no adverse effect on cell function and appears to be a suitable adjuvant agent for blood cell component collection.


Allogeneic unresponsiveness to orthotopic cardiac transplants in DL-A-identical radiation chimeras. Nine Cooperstown beagles of known DL-A genotypes were exposed to supralethal total-body irradiation and received bone-marrow allografts from DL-A-identical donors. Four to 5 months later, the resulting chimeras received orthotopic cardiac allografts from their corresponding donors of marrow. Six chimeras died of operative complications in the immediate postoperative period. The other 3 chimeras survived from 173 to 547 days; 1 dog died at 173 days as a result of right-sided heart failure, secondary to stenosis at the site of the pulmonary artery anastomosis. The other two recipients continue to be active and healthy at 545 and 547 days. The results indicate that dogs can be rendered specifically tolerant to orthotopic cardiac allografts by supralethal total-body irradiation and the transplantation of marrow obtained from the prospective allograft donor.


Viability of granulocytes obtained by filtration leukapheresis. Nylon filtration alters the morphology of human granulocytes and slightly reduces their phagocytic ability as measured by activation of the hexose monophosphate shunt (HMS) pathway of glucose metabolism. In addition, the ability of rat granulocytes to circulate in granulocytopenic rats is reduced, and this is not attributed to HMS activation during filtration. The activation of the hexose monophosphate shunt is related to the size of the particles rather than to the nature of the material.


Use of hydroxyethyl starch to improve granulocyte collection in the Latham blood processor. This report describes a practical, relatively inexpensive system of leukapheresis and plateletpheresis, utilizing the Haemonetics blood processor. The results of 236 phereses of normal donors are reported, with particular attention to an evaluation of five different anticoagulant mixtures: ACD, 2 per cent citrate in saline, and three different combinations of citrate and hydroxyethyl starch (HES). These mixtures were compared with respect to their effectiveness in the harvesting of granylocytes and platelets, respectively. The best harvest of granulocytes (3.5 X 10(9) cells per liter of blood processed) was with 6 per cent HES anticoagulated with trisodium citrate. All solutions gave about the same platelet yields (a mean of 1.6 X 10(11) per liter of blood processed). Because of its higher citrate content, ACD caused three times as many donor reactions as the other solutions. The use of HES thus permits this widely available plateletpheresis system to be used for leukapheresis with only minimal procedural modification.


The solid phase radioimmunoassay technique for hepatitis B (surface) antigen: effects of progressive modification. Serum specimens submitted for confirmation of hepatitis B antigen (HBsAg) reactivity were tested by counterelectrophoresis (CEP) and by three modifications of the solid phase radioimmunoassay "sandwich" technique (SPRIA). In addition, the specificity of each reaction was determined for two of the SPRIA test methods. One hundred seventy-five of the 688 specimens were reactive by all techniques and a further 60 ere specifically reactive by two of modifications of SPRIA. An additional five specimens were positive by a third SPRIA modification. The primary SPRIA technique, using a homologous guinea pig antibody sandwich, detected 184 nonspecific reactive samples. With the same test done at 45C with a reduced incubation time, there were only 37 nonspecific reactive samples. None of the nonspecific samples was detected when the SPRIA technique used a heterologous guinea pig-human antibody sandwich. The results provide a basis for estimating the effectiveness of the SPRIA technique for HBsAg testing.


[Zinc-iodine-osmium impregnation of giant synaptosomes obtained from the hippocampal formation of the rabbit]. The fraction of giant synaptosomes from the r. inferior of the rabbit hippocampus was studied using impregnation with zinc iodide-osmium tetroxide (XIO) reagent and electron microscopy. In this fraction, light and dark synaptosomes were observed. The reaction product was found in the clear-centered synaptic vesicles (200-400 A) as electron-dense structures of different forms and small osmiophilic particles on the vesicular membranes. Dense-cored vesicles and postsynaptic structures were not revealed with ZIO-reagent. The structures revealed with ZIO-reagent in the giant synaptosomes of the hippocampus are supposedly related to stroage of the neurotransmitter-glutamate.


[Conditions of periodic oxidation in the determination of the fluorescent PAS reaction]. The effect of temperature, pH, and concentration of potassium periodate solution on the intensity of fluorescence PAS-reaction was studied. The reaction was shown to be most specific in respect to glycogen at the temperature of oxidizer 4 degrees C (optimal time of oxidation being 10-12 hr). Similar results were also obtained at 20 degrees (optimal time of oxidation being 1.5 hr). Under these conditions the curve of dependence of the reaction intensity on the time of auramine-SO2 treatment double-stepped character which can be due to the presence of two glycogen fractions in hepatic cells. The fluorescence PAS-reaction intensity varied in significantly when the periodate concentration changed from 0.8 to 1.2% (from 0.2 to 0.8%) it gradually increased and pH -from 2.0 to 6.0.


[Differential staining of the chromosomes in transplanted ascites tumors in mice (in connection with the discussion on the existence of the NK/Ly strain)]. When studying the Q-, G-, and C-bands, the character of acrocentric chromosomes' bands was found to be varying even within the same tumor. However, the bands of the markerchromosomes A and B coincide in NK/Ly and in the Ehrlich--Lettres tumors. With the aid of staining for revealing C-bands, the acrocentric markers "cc" and "ac" common for both tumors, were first noted. This corroborates the opinion of the replacement of NK/Ly by the Ehrlice-Lettres tumor.


[Effect of lindane on the biological effectiveness of estradiol in chickens]. After exposure of sexually immature chickens to lindan in doses of 2.5 to 25.0 mg kg-1 of feeds, the effect of the insecticide on the biological activity of estradiol was investigated. The changes in the effectiveness of estrogen, expressed by an increase of the mass of the oviduct and of the level of serum calcium, were evaluated by means of comparison of animals exposed to the insecticide with a control group that had been applied only the standard hormone dose. Proofs were obtained for the fact that the application of lindan in the quantity of the mentioned doses for 14 days caused a statistically conclusive depression of the biological effectiveness of estrogens. Compared with the control group, the stimulated mass of the oviduct of animals exposed to the insecticide was smaller by 12.7 to 17.1 p. c. and the average concentration of the serum calcium decreased by 8.0 to 27.6 p. c. The results obtained indicate that the influence of lindan already in relatively low concentrations caused significant changes in the effectiveness of exogenous estrogens.


[Changes in the biological activity of androgens after the exposure of cockerels to lindane]. A biological test was performed to study the changes in the effectiveness of androgens after the administration of different doses of lindane to sexually immature cockerels. Testosterone-propionate and Stenolon were used as reference hormones. The results indicate that the administration of lindane increases the biological effectiveness of testosterone-propionate expressed through the stimulation of the growth of comb. On the other hand, the effect of the insecticide caused a statistically highly significant reduction of androgenic activity when Stenolon was used as a reference preparation. The causes leading to different manifestations of the interactions of lindane with the two hormones tested are analyzed. The paper gives further proofs that the exposure of poultry to relatively low doses of chlorinated insecticides interferes with the biological effectiveness of sex hromones.


[Age changes in the composition of immunoglobulins in cattle]. The electrophoretic and immunochemical methods as well as gelfiltration through Sephadex G-200 showed that there are no immunoglobulins in blood serum of fetuses and new-born calves. Specific proteins in a zone of gamma-globulins electrophoretic mobility which correspond to Ig M and Ig A are found in the serum of calves 3 hrs after their feeding on colostrum. The content of these proteins reaches the maximum on the third day of life but at the age of three months it lowers up to minimum. On the third month after birth in the blood serum of calves there appears a new protein in a zone of gamma2-globulins mobility which correspond to Im G-main immunoglobulin of adult animals. An assumption is advanced that the protective functions of calves of the early age are found chiefly with immunoglobulins Ig M and Ig A.


[Effect of passive immunity on subsequent vaccination against swine fever]. Studied was the effect of a serum paratyph and Aujeszky gammaglobulin injected 10, 15, and 20 days prior to the crystal-violet and the lapinized vaccine. If the animals were infected with a pathogenic swine fever virus 10 to 15 days after their treatment with serum at the rate of 0.5 cu. cm per kg, part of them died, and the remaining contracted the disease showing a protracted course. The control vaccination on the 20th day after the animals were injected with serum led to the death of all of them showing the signs of acute swine fever. Most of the pigs injected with 5 cu. cm gamma-globulin each and 10--15 days later being infected with a swine fever virus manifested the disease, however, none of them died of the disease. If, however, after treatment with gamma-globulin they were challenged on the 20th day all of them died of acute swine fever. In the immunization of pigs with a crystal-violet vaccine on the 14th day of their treatment with Aujeszky gamma-globulin, part of them developed an atypical form of the disease, while most of them did not show symptoms whatever of swine fever. Of the immunized with a lapinized vaccine some died, and the remaining part did not contract the disease at all. All this pointed to the fact that the residual antibodies against swine fever contained in the specific Aujeszky gamma-globulin could still be in a position to inhibit the vaccinal virus, and more specifically that of the lapinized K vaccine. The vaccination against swine fever should be performed 14 days after the injection of the serum paratyph and about 20 days after the injection of the Aujeszky gammaglobulin.


[Cells of rat lymphosarcoma in chambers for intravital microscopy]. In a chamber for intra vital microscopy, implanted subcutaneously in a rat, stained rat lymphosarcoma cells were applied into a preliminary formed connective tissue layer. The process of intracellular transformation of a dye and its elimination from the cytoplasm was observed. Following elimination of the dye a duplication of lymphosarcoma cells would proceed. This is a very intensive process, and after 5-7 days the connective tissue is completely substituted by tumor cells.


[Assessment of the mental and physical development of children subjected to surgery for hydrocephalus during infancy and early childhood]. Some psychoneurological problems related to the operative treatment of hydrocephalus at an early age are discussed. Under a continual observation for a period of 2 years were kept 28 children-hydrocephals who had undergone ventriculoatriostomy. Once a month they were subjected to a psychoneurological examination according to a program described in the article. During each investigation the dynamics of the psychophysiological development of the children was analyzed and the degree of their mental and physical progress determined. On expiration of 22 months 32 per cent of the children were found to be fully in keeping with the average development standards, this number remaining stable during the last 5-6 months. The author attempts to justify the need for a continuous psycho-neurological control over such surgical patients, as well as the necessity to follow a definitely set system of screening such patients for surgery, well-based on pertinent psycho-neurological criteria.


Bleomycin in testicular teratomas.

Bleomycin in testicular teratomas. Bleomycin and its combinations have a definite place in the treatment of metastatic testicular carcinomas other than seminomas and may also prove to be effective, when given as adjuvant postoperative therapy in state II disease. A study, using the combination of Bleomycin and CCNU in 17 patients with metastatic testicular teratoma, is reported. A remission rate of 59% has been observed. Only patients with a complete remission had a satisfactory remission duration. Sequential combinations, using all drugs effective in this disease, may be the most promising way to further improve therapeutic results in terms of remission rate and survival.


[Role of vitamins in the prevention of atherosclerosis]. Persons doing brain work and taking aerovit in three courses of 30 days each with intervals of 5-6 months were kept under observation over a period of 13-15 months. While improving the vitamin allowance the course-wise taking of aerovit helped normalize the lipids metabolism factors characterizing an increased predisposition to atherosclerosis, the favourable effect being traced for 4-5 months after discontinuance of the preparation ingestion. Repeated courses contributed to a further improvement of the lipoid metabolism. A tendency towards positive changes in the tonicity of the intracranial vessels was recorded, while in subjects with an increased pulse wave propagation rate along the elastic type vessels this tonicity was seen to decline. With the available data on the positive effect of aerovit upon the lipoid and protein metabolism and some clinical and instrumental findings serving as a background an inference is made that a prolonged (course-wise) application of aerovit can be of help in preventing the development of atherosclerosis.


The histological and histochemical studies on the ovary in relation to vitellogenesis in the dragonfly, Orthetrum chrysis Selys (Libellulidae: Odonata). The ovaries consist of large number of panoistic ovarioles in the last instar nymph and the adult dragonfly Orthetrum chrysis (Selys). In the nymph the vitellaria are compactly filled with the primary oocytes and the vitellogenesis takes place only in the adult stage. During vitellogenesis oocytes change widely in their shape, size and cytological organisation and their developmental stages can be divided into pre-vitellogenic, early-vitellogenic, vitellogenic, late-vitellogenic and maturation age. PAS-positive material appears first around the germinal vesicle in the early-vitellogenic stage and lateron it migrates towards the periphery. Glycogen appears in the late-vitellogenic stage. DNA is abundantly present in the nuclei of the oocytes during the pre-vitellogenic and completely absent in early-vitellogenic, vitellogenic, late-vitellogenic and maturation stages. It is observed in the nuclei of follicular epithelial cells of all the stages. RNA is abundantly present in cytoplasm of the pre-vitellogenic oocytes but lateron is gradually decreases. During the early-vitellogenic and vitellogenic stages high concentration of RNA in the follicular epithelial cells has been observed. The protein bodies appear first in the interfollicular spaces and towards the periphery of the oocytes just near the enveloping follicular epithelial cells, during the early-vitellogenic stage suggesting the formation of yolk proteins from the haemolymph. In Orthetrum chrysis the sudanophilic bodies appear first in the follicular cells and then lie in the peripheral region of the oocytes suggesting the incorporation of yolk lipid either from the follicular epithelium or from the haemolymph through the follicular epithelium. The phospholipids are synthesised in pre-vitellogenic to the late-vitellogenic stages. In the late-vitellogenic stages the phospholipid granules are present abundantly in the follicular epithelium while in the maturation stage they disappear suggesting their utilisation in the formation of membranes like vitelline and chorion. The neutral fats are present in the form of large number of droplets in the oocytes during the maturation stage.


Cellular composition of the islets of langerhans in the himalayan toad, Bufo melanostictus (Schneider): a light microscopical study. The pancreatic islet tissue of Bufo melanostictus, investigated by differential staining techniques, is generally condensed in the anterior and middle regions, and contains distinguishable islets of various size, shape and or irregular configuration. Histologically, 3 distinct cell types have been identified: B, A1 and A2. Various tinctorial characteristics of B cells reveal that they correspond to the insulin producing B-cells of other vertebrates. The A cells are a few in number, some of which definitely show positive argyrophilia (= A1). A few isolated A- and B-cells are found scattered in the exocrine tissue. A conspicuous feature of several B-cells in some specimens of Bufo melanostictus is the presence of vacuoles of varying size.


Variants in the histochemical patterns of lipids in the hepatopancreas of Scylla serrata (Forskal) (Brachyura, Decapoda). The highest concentrations of phospholipid, neutral lipid and fatty acids were observed in the R cells and connective tissue of the hepatopancreas of Scylla serrata. The basal parts of the B cells and apical parts of the cells lining the main duct also showed moderate presence of these substances. The E cells however, except at their cell membranes were found to be devoid of lipids. F cells on the other hand exhibited lipoid complexes. Considerable reduction in the staining intensity of fatty acids were noticed 4 h after the bilateral ablation of eyestalks, neutral lipid undergo depletion 24 h after the operation whereas phospholipid reserves increase 48 h after the eyestalk removal. A fall in the quantity of neutral lipid and phospholipid was conspicuous when eyestalk extract was injected into normal or destalked crabs. From the present data it appears that R cells and connective tissue form major sites of lipid storage and in an intermolt animal eyestalk factor(s) may have an important role in the control of lipid metabolism.


[Measurements of cell nuclei in blood smears from frogs]. In the blood-spread of frog size and extinction of nuclei of erythrocytes, thrombocytes and neutrophile leucocytes with segmental nucleus were investigated. The substance of nucleus was made visible by means of colouring in according to May-Grünwald and the DNA-content in according to Feulgen-reaction. The thickeness of layer and relief of the spread were recorded and measured with a touch-equipment. The nuclei and segments of nucleus, which are flat disks in the dry spread (nuclei of thrombocytes are about 0.75 mum thick), are considered to be rotational ellipsoids in the fluid blood; volume, surface and quantity of DNA are calculated. The erythrocytes have the smallest. the leucocytes the largest quantity of DNA in the nucleus. The concentration of the substance of nucleus and of the DNA increases with the dryness of the spread, to about the sixfold value at the nuclei of thrombocytes.


[Serum levels of alpha-2-macroglobulin, ceruloplasmin, transferrin, alpha-1-antitrypsin and complement (beta-1-C) before and following 3- and 6-day injections of D-penicillamine in man]. The serum levels of alpha-1-antitrypsin, ceruloplasmin, transferrin, alpha-2-macroglobulin and complement (beta-1-C) were measured by the radioimmunodiffusion method of Mancini before treatment, after three days treatment with D-penicillamine (1,00 g. i. v. daily) and after treatment for 3 days with 2 x 1,00 g i. v. D-Penicillamine. The levels of alpha-2-macroglobulin, coeruloplasmin and transferrin dropped significantly after 3 days' treatment. Doubling the dosage for 3 days induced a further drop in transferrin and ceruloplasmin. The alpha-2-macroglobulin was not further influenced. The levels of alpha-1-antitrypsin and complement were not influenced by D-Penicillamine treatment. D-penicillamine, alpha-1-Antitrypsin, alpha-2-macroglobulin, transferrin, ceruloplasmin, complement (beta-1-C).


[On the effect of some inhibitors of proteinbiosynthesis on activity and learning behavior of goldfish (Carassius auratus auratus L)]. Concerning the problems of learning and memory there is a distinction of a short term memory (STM) and a long term memory (LTM). It is supposed that the STM is an electrical phenomenon, whereas the LTM depends on material changes. The assumed materials are RNA, proteins, lipids, amines etc, and the primary carrier of the information is DNA. But there is a discrepancy: learning-specificity is based on environmental changes, but not the structure of DNA. For the investigation of this, we trained goldfish in a shock-free task to take food from coloured cups under the influence of inhibitors of the proteinbiosynthesis. There was no inhibition on memory-processes in our experiments.


[The spontaneous NBT-test in granulocytes of patients with juvenile rheumatoid arthritis]. 30 patients with definite seronegative juvenile rheumatoid arthritis and 3 patients with tendosynovitis and unclear arthralgia respectively were examined by the nitroblue tetrazolium (NBT) test (46 tests). A positive result could be obtained in 72 per cent of the examinations. Patients with a joint manifestation of juvenile rheumatoid arthritis showed relatively more positive results than patients with Morbus Wissler and the transition forms of juvenile rheumatoid arthritis. Tendosynovitis and unclear arthralgia were negative in the NBT test. The NBT test is not useful for the differentiation of bacterial-septic and rheumatoid-inflammatory joint diseases.


The adaptation of fluorescence microscopy for Nocardia asteroides: a preliminary report. The Fluorescence Microscope is extremely effective in revealing the presence and morphology of Nocardia asteroides. This has been effectively demonstrated when the Auramine Fluorochrome was decolourized with 1% H2SO4 and the interval of decolourization was limited to two minutes and thirty seconds. When Nocardia asteroids was seeded in sputum found to be negative for acid-fast organisms and in greatly diluted quantities after one hour intervals, its presence was still revealed with the Flourescence Microscope. The normally employed Kinyoun technique of acid-fastness was seen to be less effective under the same conditions.


Antigenic subgroups of hepatitis B (surface) antigen in Berlin (West) and their epidemiological aspects. Sera of 334 hepatitis B patients and 118 sera of HBsAg carriers were tested for the distribution of the subgroups ad and ay by means of rheophoresis technique or solid phase radiommunoassay. The distribution of subgroups revealed a typical pattern. The ad-antigen determinant was more frequent in hepatitis B patients, blood donors and hemodialysis patients whereas the ay determinant was detected in drug users, guest workers and juveniles at a considerably higher percentage. The difference in distribution is discussed as an epidemiological phenomenon of hepatitis B virus, dependent on environmental factors characteristic for certain groups of the population. The routine testing for HGsAg subgroups is recommended as a valuable epidemiologic tool.