Studies on rheomelanins. IV. The apparent occurrence in vivo of rheomelanins in human blood. Paper chromatograms of the rheomelanins made earlier in this laboratory in human plasmas during incubation with each of the catecholamines or with L-dopa had yellow-green fluorescence in ultraviolet light of 366 nm (Hegedus & Altschule, 1970). In the present studies, a yellow-green fluorescent spot was found in each paper chromatogram. All of these spots were eluted and their excitation and emission spectra were recorded and compared to one another. The rheomelanins made from the catecholamines, L-dopa, catechol or from mixtures of these in human plasmas during incubation were chromatographed twice on paper with two different solvent systems. These artificial rheomelanins in vitro and the apparent in vivo rheomelanins present in plasmas moved together during the two chromatographies. These yellow-green fluorescent compounds were eluted as one spot after the second chromatography. This mixture produced high intensity excitation and emission spectra closely similar to the low intensity excitation and emission spectra of the apparent in vivo rheomelanins of unincubated human plasmas treated the same way without any chemical under nitrogen. The RF-values of the various rheomelanin spots after each chromatography were closely similar. These above results were also obtained with other solvent systems. It appears therefore that rheomelanins are formed in vivo in the human body.


Partial purification and properties of rat-heart adenosine kinase. The activity of myocardial adenosine kinase (E.N. 2.7.1.20) in a number of species was assayed. Rat heart contained the highest specific activity. From this source adenosine kinase was purified in a simple way 80-fold, until it was free of adenosine deaminase activity. A molecular weight of about 39 000 was measured. NSC 113939 (1), NSC 113940 and 8-azaadenosine inhibited myocardial adenosine kinase. Dipyridamole stimulated the enzyme at high adenosine levels, and inhibited at low substrate concentrations. A number of divalent cations could (partially) substitute for Mg2+. The optimal concentration of MgCl2 or MnCl2 was about 0.5 mM; concentrations exceeding 1 mM inhibited severely. An apparent Km for ATP of 0.1 mM was measured, whereas an apparent Km for adenosine of 0.5 muM was was found. The latter increased to 3.3 muM, when dipyridamole was added. Replacement of ATP by GTB or ITP increased the activity, and UTP and CTP were inferior as a phosphate donor.


Blood levels of cyclic AMP, thyroxine, uric acid, certain metabolites and electrolytes under heat-stress and dehydration in the pigeon. The effect of heat stress and dehydration on certain blood parameters were studied in the pigeon. Pigeons exposed to heat-stress and dehydration showed higher haematocrit values. Serum cyclic AMP levels, both with and without corrections made for difference in haematocrit, showed a more than two-fold increase in heat-stressed and dehydrated pigeons. Serum triglyceride level (before and after correction for haematocrit difference) in the experimental birds dropped to less than half that in the control birds. Total serum thyroxine (T4) did not show significant change in the experimentals (either with or without correction for haematocrit), although bound T4 and T3 index showed a statistically significant drop in the experimental group, after the correction for haematocrit. Total serum protein and albumin values after correction for haematocrit difference showed significant decreases in experimental birds. In pigeons exposed to heat-stress and dehydration, serum sodium increased while potassium dropped (with and without haematocrit correction). Heat stress and dehydration did not have any significant effect on serum levels of total cholesterol, uric acid, sodium, potassium, calcium and magnesium.


Translation of mRNA for glutamate dehydrogenase and spectrophotometric procedure to follow the enzyme biosynthesis. Heterogeneous poly (A)-mRNA fraction was isolated from rat liver microsomes using phenol-chloroform extraction, millipore filtration and poly (U)-agarose affinity chromatography. Obtained fractions were characterized with respect to their secondary structure and poly (A) content. Isolated poly (A)-mRNA fraction contained high template activity for glutamate dehydrogenase in cell-free systems with microsomes or polysomes. A spectrophotometric procedure to follow enzyme biosynthesis was also developed.


Change of the activities of angiotensins I and II on the vascular smooth muscle by crude kallikrein. Incubation in vitro of angiotensin I (A I) with crude kallikrein induced a potentiation in the response to decapeptide of the isolated continuously superfused rabbit aorta. Crude kallikrein when incubated with angiotensin II (A II) caused a decrease in response to octapeptide of the same assay tissue. Converting enzyme inhibitor, SQ 20881, produced a competitive inhibition in the response to A I preincubated with crude kallikrein but did not alter the inhibitory effect of the enzyme on A II response. Pure kallikrein did not induce any change in the responses to both peptides when used at the same concentrations. The competitive inhibitor of A II (N,N-dimethyl) Gly1-Ile5-Ile8-angiotensin II (DMGIA II), abolished the effects of both A II- and A I-preincubated with crude kallikrein. From these results it was concluded that crude kallikrein-induced potentiation in the response to A I of the aorta is probably due to the conversion of decapeptide to octapeptide by an enzyme fraction in crude kallikrein preparation. These results also indicate that crude kallikrein (Padutin) is not a pure enzyme preparation and probably contains some other enzyme fractions which are responsible from the changes of the vascular activities of angiotensin-peptides.


Effects of the number of DNP groups on dinitrophenylated human serum albumin upon the induction of immunological memory for the dinitrophenyl determinant. Immunological memory to the DNP determinant is shown to be a positive correlate of the primary anti-DNP response and of the extent of DNP substitution in the priming antigen. Antigen dose-response curves for immunological memory first increase with antigen dose to reach a maximum and then decrease in the high dose region. Increasing the extent of DNP substitution in the priming antigen shifts the dose-response curves to lower antigen doses and promotes the antigen's ability to induce immunological memory to a level greater than that expected by its ability to induce a primary response. The findings are interpreted in terms of postulated effects of degree of DNP substitution on the binding of antigen to cell receptors.


[A system of computer pre-treatment of electrocardiographic data]. A system of analogue pretreatment aimed at analysis of the arrhythmias in real time is presented. It is based on the separate detection of the P wave and the QRS complex, and on the use of a bipolar electrically conducting catheter introduced into the auricle of the right atrium. Such an arrangement is relatively insensitive to adventitous activity, both in the high and low frequency range, and provides on four separate channels impulses representing the P wave, the QRS complex, wide QRS complexes and premature QRS waves. In all cases, the atrial channel transmits all synchronised signals originating from atrial activity without any loss of information. This apparatus has been tried out with various arrhythmias and, if integrated, into a system of automatic detection, it should bring about a considerable saving in time and memory storage space.


[Structures of oncornaviruses of C-typ-negativ staining technique by electron microscopy (author's transl)]. Typ-C viruses of bovine enzootic leucosis and other typ-C viruses were demonstrated by negativ staining technique and also the RNA-protein complex respectively the nucleocapsid of helical symmetry in the shape of coral bead. Now the well defined morphology of the typ-C viruses enable to identificate to be used in electron microscopy practice.


"Runde" viurs, a coronavirus-like agent associated with seabirds and ticks. From 206 I. uriae collected in the seabird colonies at Runde, Norway, two identical virus strains demonstrating no antigenic relationships to major arbovirus groups were isolated. The new strains demonstrated a corona-virus like morphology, haemagglutinated chicken red cells and were sensitive to sodium desoxycholate. Multiplication with CPE was demonstrated in BHK 21/c13 and BSC-1 cells, and without CPE in Vero and GMK cell cultures. The mouse pathogenicity was relatively low. In gel precipitation three to five specific lines were seen. Precipitating antibodies have been found in seabird species commonly infested by I. uriae. The ecological cirucmstances of the isolates indicate an earlier unrecognized arbovirus circulating between seabirds and I. uriae. This corona-like virus has been tentatively termed Runde virus.


[Morphological characteristics of acute alcoholic hepatitis]. Liver biopsies from 60 patients with acute alcoholic hepatitis (AAH) developing against the background of steatosis, chronic hepatitis and cirrhosis were studied histologically, histochemically and electron microscopically. AAH is characterized by necrosis of hepatocytes with deposition of alcoholic hyalin, obesity of the organ, and polymorphonuclear leukocyte infiltration. Hyperplasia of the smooth endoplasmic reticulum, the appearance of megamitochondria, and an increased amount of peroxisomes reflect the participation of MEOS and the catalase system in alcohol metabolism with a progressive decrease in the activity of alcoholdehydrogenase. Acute alcoholic hepatitis is a connecting link between steatosis and cirrhosis of the liver in which the accompanying autoimmune mechanism and microcirculation disorder followed by activation of lipofibroblasts are conducive to the progression of the pathologic process.


The current approach to cancer treatment. The therapeutic armamentarium in the treatment of cancer is now expanded to include surgery, radiotherapy, chemotherapy and immunotherapy. The aim is to achieve cure where possible. For some cancers the concept of cure can be questioned, and for many patients, cancer must be regarded as a chronic disease. A vigorous approach to palliation, particularly in regard to special problems such as hypercalcaemia, yields good dividends. Human values remain of paramount importance, and active antitumour treatment with attendant side effects must not be continued where there is no expectation of worthwhile response.


Single channel characteristics of some synthetic gramicidins. The synthesis of two new variants of gramicidin is described. It is shown that the changes in the aromatic side groups do not influence the single channel conductivity. Experiments in which solutions having different molarities on the two sides of the bilayer lipid membrane are described and their results compared with a rate theory analysis. It is concluded that the gramicidin pore contains approximately 10 equal potential maxima.


Diffusion and partitioning of a pesticide, lindane, into phosphatidylcholine bilayers. A new fluorescence quenching method to study chlorinated hydrocarbon-membrane interactions. Chlorinated hydrocarbons, such as the pesticide lindane (gamma-hexachlorocyclohexane), quench the fluorescence of carbazole. The observed quenching is a result of the molecular contacts which occur upon diffusional collisions. Because the amount of quenching depends upon the collisional frequency between carbazole and pesticide, this phenomenon provides a measure of both the diffusional rate of lindane and its local concentration. The carbazole fluorophore is localized within phosphatidylcholine bilayers by cosonicating the lipid with a newly synthesized phospholipid, beta-(11-(9-carbazole)-undecanoyl)-L-alpha-phosphatidylcholine. Using this probe in dimyristoyl-L-alpha-phosphatidylcholine vesicles, and the above mentioned quenching phenomena, we determined the lindane diffusion rate within the bilayer to be 5.7.10-7 cm2/s at 37 degrees C. Measurement of the apparent quenching constant at various dimyristoyl phosphatidylcholine concentrations yielded a lipid-water partition coefficient for lindane of 9500, which is in agreement with the value of 8980 obtained by our equilibrium dialysis experiments. Vesicles of dimyristoyl-L-alpha-phosphatidylcholine become saturated with lindane at a pesticide to lipid molar ratio of approx. 0.28. These results demonstrate the possibility of using the quenching of carbazole fluorescence to investigate the transport and partitioning of pesticides within biological membranes. This ability should prove useful in studies of the interactions of chlorinated hydrocarbons with cell membranes.


NMR observation of gramicidin A' in phosphatidylcholine vesicles. Dimyristoyl phosphatidylcholine was prepared with perdeuterated hydrocarbon chains and sonicated into bilayer vesicles together with gramicidin A'. The 1H NMR resonance from the tryptophan residues in the gramicidin has a linewidth of approximately 80 Hz, indicating significant local mobility for these residues. Paramagnetic lanthanides added to the aqueous medium cause a chemical shift of this signal indicating that some of the tryptophans may be located in the interfacial region of the bilayer.


Protamine messenger RNA from rainbow trout testis contains the nucleotide sequence A-A-U-A-A-A in an untranslated region. Full-length, complementary DNAs were prepared to rainbow trout protamine mRNA using reverse transcriptase and were labelled during synthesis by the replacement of dATP by [alpha32P]dATP or dTTP by [alpha32P]dTTP. The 32P-labelled protamine complementary DNAs were digested with T4 endonuclease IV. Fragments from the digests were separated in two dimensions, and those discrete fragments which could be identified from both the A-labelled and T-labelled complementary DNAs were subjected to sequence analysis. The sequences described here all arise from the non-coding region. One pentadecanucleotide contained the sequence A-A-U-A-A-A which has been reported by Proudfoot and Brownlee ((1976) Nature 263, 211-214) to occur in the noncoding regions of six other eukaryotic mRNAs.


Microbial secondary metabolites with potential use in cancer treatment. (Plasmid involvement in biosynthesis and compounds. Antibiotics and small molecular enzyme inhibitors produced by micro-organisms are microbial secondary metabolites which have no obvious function in the growth of microorganisms. Involvement of plasmids in their biosyntheses has been studied and the structure of the microbial world where various compounds with widely varying structures have been found has been shown to be due to the presence of a great variety of plasmids. On the basis of this structure of microbial world, the author has found the microbial products which have various activities useful in treatment of cancer or in suppression of carcinogenesis. In this paper, the author reviewed his following studies: the plasmid involvement in biosyntheses of bioactive microbial secondary metabolites; the mechanism of therapeutic effect of bleomycin, and the development of useful analogs and derivatives of bleomycin and anthracyclic antibiotics; the screening method to find compounds affecting immune system, and the action of bestatin in enhancing delayed-type hypersensitivity and therapeutic effects of anticancer agents; enzyme inhibitors with potential utility in treatment or prevention of cancer.


Acute phase reactant proteins in chronic leukaemia. The levels of haptoglobin, alpha1 antitrypsin and alpha1 acid glycoprotein are moderately raised in chronic leukaemias. In CGL the level of haptoglobin and acid glycoprotein show the highest correlation with cell number, whilst no such correlations occur in CLL or CMML. There does not appear to be a relation between blood lysozyme levels and the levels of antiprotease (alpha1 antitrypsin and alpha2 macroglobulin).


[Influence of five commercial heparin preparations on the collagen induced platelet aggregation (author's transl)]. The ADP- and collagen-induced aggregation of platelets in PRP of healthy probands was tested in vitro in the presence of 4 heparin preparations (Novo, Vitrum, Liquemin, Haemoderivate) and of a heparinoid (Eleparon). None of the preparations caused a significant direct aggregation. The collagen-induced aggregation was inhibited by all mentioned preparations; thereby the heparins proved to be effective approx. to the same degree whereas the heparinoid proved to be significantly less effective. The ADP-induced aggregation was potentiated by all preparations. A considerably different sensitivity to heparin could be observed in the single PRP's. A special method was found out to exclude the influence of the heparin-sensitivity of the PRP's to compare the special effects of the heparins.


[Association of chemotherapy and radiotherapy in the treatment of advanced cancer of the head and neck]. At the Head and Neck and Roentgentherapy Services of Fundaçao Antonio Prudente, São Paulo, Brazil, 38 patients with advanced cancer of the oral cavity and oropharynx, were divided into two groups: the first, submmited to treatment with Bleomycin and Linear Acelerator, and the second to isolated irradiation. Both groups received 7 000 rads in 32 days, and to the association group, Bleomycin was administered in dosis of 300 mgs. The authors concluded that, that Chemo-Radiotherapic association didn't increase the survival rate.


The inhibition of histamine release from rat peritoneal mast cells by non-steroid anti-inflammatory drugs and its reversal by calcium. 1 The non-steroid anti-inflammatory drugs, indomethacin, flufenamate and meclofenamate, inhibited the release of histamine from rat peritoneal mast cells induced by pharmacological or immunological challenge in vitro. 2 Anti-inflammatory steroids had little effect on histamine release from the mast cells. 3 Th inhibition of histamine release by the aspirin-like drugs was not prevented by incubation with glucose, unlike the inhibition of 2,4,dinitrophenol or antimycin-A. This suggests that the non-steroid anti-inflammatory compounds do not act by preventing the energy production from oxidative metabolism, required for histamine release. 4 The inhibition of the calcium ionophore A23187-induced histamine release by the aspirin-like drugs was reversed by an increase in the calcium concentration of the incubation medium. 5 The results suggest that the non-steroid anti-inflammatory compounds inhibit histamine release by actions on calcium influx into the mast cell, or on calcium mobilization or utilization within the mast cell.


The relationship between energy metabolism and the action of inhibitors of histamine release. 1 Dextran-induced release of histamine from rat mast cells was inhibited equally in complete and glucose-free Tyrode solution by doxantrazole (0.03-3 micronmol/l), theophylline (0.1-3 mmol/l) and dicumarol (0.01-10 micronmol/litre). 2 Doxantrazole (3 micronmol/l), theophylline (3 mmol/l) and dicumarol (10 micronmol/l) did not reduce the adenosine 5'-triphosphate (ATP) content of mast cells in glucose-free medium. Higher concentrations of dicumarol (56-100 micronmol/l) markedly reduced the cellular ATP content. This reduction was reversed by glucose. 3 Papaverine was a more potent inhibitor of histamine release from mast cells incubated in glucose-free solution than in complete Tyrode solution (dose-ratio = 20). Like antimycin A (L MICRONMOL/L), PAPAVERINE (3 MICRONMOL/L) CAUSED A DEPLETION OF MAST CELL ATP that was greater in the absence (85%) than in the presence (25%) of extracellular glucose. 4 These results suggest that dicumarol, like doxantrazole and theophylline, inhibits histamine release without affecting mast cell energy metabolism. In contrast, papaverine probably inhibits release by depleting ATP that is required for exocytosis. 5 Inhibition of histamine release by dibutyryl cyclic adenosine 3,5'-monophosphate (1-3 mmol/l) was significantly greater when cells were incubated in complete rather than in glucose-free medium.


Surgery, venous thrombosis and anti-Xa.

Surgery, venous thrombosis and anti-Xa. Deep venous thrombosis is a common and unpredictable complication of surgery. In this study it is proposed that patients who develop this complication may be predicted by a low preoperative level of a naturally occurring inhibitor of coagulation, anti-Xa. Two groups of patients were investigated. Women taking the oral contraceptive pill had lower preoperative anti-Xa levels than their non-pill controls (P less than 0.01) and in addition had a significantly higher incidence of deep venous thrombosis (DVT) following emergency surgery (P less than 0.05). In 90 patients undergoing total hip replacement, the mean preoperative anti-Xa level of those patients who developed DVT was significantly lower than those who did not (P less than 0.001). Ninety-four per cent of patients with a preoperative anti-Xa level of less than 80 per cent developed DVT. The effect of low dose heparin on anti-Xa was studied. The results suggest a mechanism for the cause of postoperative thrombosis which also permits selection of individual patients who will develop this complication.


The prospective evaluation of tumour-associated antigens for the early diagnosis of pancreatic cancer. An unselected series of 136 patients suspected of having a major intra-abdominal pathological condition had blood drawn and stored as serum. A definitive histological diagnosis was made in all cases, usually by laparotomy. The serum was analysed with coded samples for carcino-embryonic antigen, alpha-fetoprotein and pancreatic oncofetal antigen and all recorded levels above the upper limit of normal for that test were taken as a positive result. Results were analysed to give the predictive value of a positive test and a negative test for each antigen. A negative pancreatic oncofetal antigen test was found to be useful in excluding a diagnosis of pancreatic cancer.


Alpha-adrenergic blockers in prostatism. Thirty patients awaiting prostatectomy with no history of cardiovascular disease were selected for a trial of alpha-adrenergic blocking agents. Twenty-seven patients completed the trial, 19 of them with unpleasant side effects. Eighteen of them found their symptoms were improved and 10 showed urodynamic improvement.


Prostatic enlargement associated with symptomatic and asymptomatic gonorrhoea in adolescent men: An argument for routine rectal examination. Ten adolescent men presented with prostatic enlargement and positive anterior urethral cultures for gonococci. Six of them had urethritis, but four were asymptomatic; the prostate is therefore implicated as a reservoir for the asymptomatic carrier and it is strongly recommended that the rectum should be included in the routine physical examination of sexually active adolescent men.


Axonal transport of organelles visualized by light microscopy: cinemicrographic and computer analysis. Rapid movements of intra-axonal organelles in acutely isolated single myelinated fibers from bullfrog sciatic nerve were visualized by dark-field microscopy. The movements were recorded by cinemicrography, and analyzed by computer-based methods. The movements are saltatory and bidirectional, but each particle moves mainly in a single direction. For more than 90% of the particles, the predominant movement direction is retrograde, i.e. toward the cell body. Quantitative measurements on a variety of parameters of the organelle movements are presented. Different particles in the same axon show a broad range of mean speeds. The average mean speed of movement in the retrograde direction at 28 degrees C was 1.08 micrometer/sec (S.D. - 0.41), equivalent to an axonal transport rate of 93 mm/day. Disperse distributions were also found for other parameters such as the instantaneous velocities of individual particles. Quantal velocities, periodic movement patterns, and specific 'channels' were not detected. When the data from a population of particles is treated statistically, the average mean speed, the distribution of velocities, and other statistical parameters are found to be similar in different axons studied at the same temperature. Direct microscopical observation of axonal organelle movement is a technique which provides information about axonal transport which is different from and complementary to that obtained from enzyme accumulation of radioactive tracer methods.


The intra-axonal transport of polypeptide H: evidence for a fifth (very slow) group of transported proteins in the retinal ganglion cells of the rabbit. We have determined that a genetically polymorphic polypeptide (H, molecular weight approximately equal to 195,000) of the rabbit nervous system is transported down the retinal ganglion cell axons at a velocity of 0.7-1.1 mm/day. The H-polypeptide and probably at least two additional polypeptides (molecular weights approximately 145,000 and 73,000) therefore compose a group of intra-axonally transported proteins which moves more slowly than the 4 groups previously described in these neurons. The polypeptides of this fifth group are similar in molecular weight to certain polypeptides transported slowly in other mammalian neurons.


Intraaxonal transport of Herpes simplex virus in the rat central nervous system. Light and electron microscopic observation 3--4 days after microinjection of Herpes simplex virus (HSV) into the left neostriatum of rat demonstrated the following results. (1) Virus labeled nerve cells were found in the ipsilateral substantia nigra; a large number of infected neurons were in the zona compacta and some were in the zona reticulata. No virus infection was evident in the contralateral side. (2) Virus labeled neurons were found in the cortex, a greater number ipsilaterally than contralaterally, and in the dorsal raphé nuclei. Cortical microinjection of HSV led to infection of some cortical cells but no neostriatal cells. We conclude, therefore, that spread of the virus to the cortex, the substantia nigra and the dorsal raphé following neostriatal injection was by retrograde axonal transport. (3) The left neostriatum, where HSV was injected, showed a surprisingly small number of virus infected neurons. The infected neurons were mostly the large neurons; the majority of medium sized neurons were well preserved. There was massive degeneration of nerve terminals throughout the neuropil. Most of these degenerating nerve terminals are considered to be afferent fibers.


A potential screening technique for neurotransmitters in the CNS: model studies in the cat spinal cord. This paper describes a potential screening technique for neurotransmitters in the CNS. The method uses the injection of small volumes of high specific activity radioactive transmitter precursor substances into regions of physiologically identified neuronal cell bodies, and the later identification of the substances transported down axons to target tissues. Experiments were performed in motoneurons in the cat spinal cords to test the feasibility  of method. Tritiated choline, glutamate, tyramine and tryptophan were pressure-injected into the ventral horn using glass micropipettes that were adapted to allow similtaneous physiological recording and injection. Only tritiated acetylcholine, two unidentified choline metabolites and a small amount of choline were found in the motor axons. The acetylcholine migrated at a rate of greater than 24mm/day and the movement was blocked by colchicine. The spread of isotope from the injection site was measured by a direct chemical method and by autoradiography, and was found that isotope spread1-2 mm from the injection site. One unexpected finding in the autoradiographs was that the motoneurons were selectively labelled following choline injections.


Electron microscopy of rapid identification of animal viruses in hematoxylin-eosin sections. Routine hematoxylin-eosin stained, paraffin sections were processed for electron microscopy, using a rapid method for localization of animal viruses. Formalin fixation was effective in preserving DNA as well as RNA viruses, however cellular fine structural details and organelles were not well preserved. The procedure is useful for morphological recognition of viral groups and as a rapid diagnostic aid for identifying viral disease.


Heparin and sulfated mucopolysaccharides--a micro system for quantitative differentiation and determination. Heparin (Hep), hyaluronic acid, chondroitins (sulfate) A, B, and C, and heparins (sulfate) A, B, C, and D were subjected to microelectrophoresis in barbital-agarose gel, fixed with cetylpyridinium chloride and stained with toluidine blue. The optical densities of the resulting bands were compared with optical densities obtained upon reaction with azure A in aqueous solution and with the carbazole reagent. A linear relation was obtained between optical density and concentration of purified sulfated mucopolysaccharide (SMP). Less than 1 microgram of Hep and 2 microgram of other SMPs are required for measurement by electrophoresis, while about 30 microgram of each is required with the carbazole reagent. The optical density of a mixture of SMPs was equal to the sum of the densities for the individual SMPs upon microelectrophoresis. It was demonstrated that the individual SMPs in mixtures were distinguishabed by reaction with specific enzymes and by changes in migration in agarose with barbital, phthalate, ethylenediamine, or propanediamine buffers, permitting ready demonstration and quantitation of various SMP species. Examples are shown of the application of the procedure to measure the total SMPs and individual SMPs in tissue extracts. The method is sensitive, reproducible, flexible, and measures quantities 1/30th of those measured colorimetrically, yet is relatively unaffected by protein, carbohydrate, or inorganic electrolytes.


Treatment of resistant ascites by continuous ultrafiltration--reinfusion of ascitic fluid. Eight patients with diuretic-resistant ascites due to cirrhosis were treated by reinfusion of concentrated ascitic fluid. In 11 procedures, with a mean duration of 21.9 hours, weight loss averaged 14.8 kg. Complications during reinfusion included septicemia in 1 procedure, left-sided heart failure in 5, pyrexia in 7 and coagulation abnormalities in 10. Ascites recurred within 2 months after reinfusion in all but one patient. Although this technique is an efficient and inexpensive method of treatment of ascites, it does not appear indicated in patients with cirrhosis and resistant ascites in view of the possibly serious complications associated with reinfusion and the poor long-term results.


Clinical pharmacologic and therapeutic studies of bleomycin given by continuous infusion. The clinical toxicology, clinical pharmacology, and therapeutic effects of bleomycin given by continuous intravenous infusion were studied in patients with far-advanced unresectable cancer. The toxicity of bleomycin given by that schedule was qualitatively and quantitatively the same as when it was given by daily intravenous "push"; mucocutaneous toxicity occurred regularly after 7-11 days of infusion. Careful monitoring of pulmonary function revealed minor changes in Total Lung Capacity and Pulmonary Diffusion Capacity in nearly all patients; however, overt pulmonary toxicity occurred in only six patients (5%). 111Indium-labeled bleomycin was used to follow blood levels of bleomycin; it correlated well with the levels determined by microbiologic assay and could be measured at levels lower than could be determined by bioassay. Useful therapeutic responses were seen in a variety of tumors; 30% patients with very far-advanced carcinoma of the cervix demonstrated CR or PR, an incidence higher than has been seen with other regimens. Sixty-nine percent of patients with disseminated germ cell neoplasms of the testis, refractory to bleomycin given by conventional dose schedules, have attained partial remission through the continuous infusion of bleomycin.


Cellular localization of alpha-fetoprotein and human chorionic gonadotropin in germ cell tumors of the testis using and indirect immunoperoxidase technique. An immunohistologic study of 21 patients with germ cell tumors of the testis with measured serum levels of chorionic gonadotropin (HCG) and alpha-feto protein (AFP) was undertaken to correlate the various types of neoplasms with the presence of these tumor markers in the tissue and serum. AFP was demonstrated in mononuclear embryonal cells within embryonal carcinoma and endodermal sinus tumor. HCG was identified within syncytiotrophoblastic giant cells, frequently in association with embryonal carcinoma, and rarely with endodermal sinus tumor and seminoma, as well as in the syncytiotropho-blastic component of choriocarcinoma. Eighteen of the 21 patients (86%) had elevated tumor markers in their serum; serum HCG alone was elevated in five (24%), AFP alone in five (24%) and both were elevated in eight (38%). There was tissue localization of HCG in 12 of the 13 patients (92%) with elevated serum HCG while AFP was identified in the tumor in eight of the 13 patients (53%) with elevated serum AFP levels. Based on these findings, a tentative immunohistologic classification of germ cell tumors utilizing AFP and HCG is proposed. Thus, embryonal carcinoma, adult type, is frequently associated with both AFP and HCG, endodermal sinus tumor with AFP and choriocarcinoma with HCG, whereas pure seminoma and teratoma are unlikely to be associated with either marker.


The identification of individuals at high risk for large bowel cancer and the application of preventive measures in their management: chromosome studies in Gardner syndrome and familial polyposis. The significance of chromosomal markers in linkage and assignment has been pointed out. The usefulness of linkage in disease prevention is discussed with the limitations of this approach.


Surface antigenic characteristics of human glial brain tumor cells. The surface antigenic characteristics of human glial brain tumor (HGBT) cells were studied by complement-dependent cytotoxic antibody assays and indirect membrane immunofluorescence. Eight permanent, well-characterized cell lines derived from human gliomas were used for analysis with antisera raised by hyperimmunization of nonhuman primates (Macaca fascicularis) with glioblastoma multiforme tissue or established HGBT cells lines. Exhaustive absorption of these antisera to remove predominantly antispecies activity rendered HLA nonreactive "preabsorbed" antisera, which reacted with a large panel of gliomatous and nongliomatous human tumor cells; 1 carcinoma, 2 sarcomas, 2 melanomas, 1 neuroblastoma, and 8 HGBT cell lines. Four lymphoblastoid lines and 2 carcinomas were unreactive. After further absorption with a human osteogenic sarcoma cell line, the antisera demonstrated significant levels of reactivity for 8 tested HGBT cell lines and no longer reacted with the nongliomatous cultured tumor cells lines. Therefore, extensive absorption of nonhuman primate anti-human glioma sera removed all activity for the nongliomatous cell lines tested, but it left significant reactivity against a glial tumor cell line-associated antigen(s) present on all 8 human glioma cell lines tested.


Cell cycle-related changes in nuclear chromatin of stimulated lymphocytes as measured by flow cytometry. Flow cytometric techniques have been developed to assay lymphocyte stimulation as reflected by the increase in the cell transcriptional activity and cell progression through the cell cycle. The metachromatic fluorescent dye, acridine orange, is used to (a) stain DNA and RNA differentially in individual cells, and (b) stain nuclear chromatin after removal of cellular RNA BY RNase and cell pretreatment at acidic pH. Stimulated cells with diploid DNA content (G1) have an increased content of stainable RNA that makes it possible to distinguish them from nonstimulated (G0) cells. G0 cells can also be distinguished from G1 cells based on differences in stainability of their nuclear chromatin after treatment with acid. Mitotic indices can be scored automatically, inasmuch as the metaphase chromatin stains differently than does chromatin in the interphase cells. Altogether, the numbers of cells in the G0, G1, S, G2, and M phases may be obtained rapidly and with great accuracy. The cell transciptional activity can be correlated with changes in nuclear chromatin (e.g., during the transition from G0 to G1). The two independent techniques may also prove to be useful in recognizing and quantitating noncycling cells in other cell systems. The possible mechanisms responsible for differential stainability of nuclear chromatin in cells at different phases of the cell cycle are discussed.


In vitro immunization against human tumor cells with tumor cell fractions. Tumor cell fractions isolated from tumor lines SH-3 (breast carcinoma) and RPMI-7932 (malignant melanoma) by differential centrifugations were capable of transforming lymphocytes into cytotoxic effector cells. Lymphocytes cultured alone in human AB plasma did not become cytotoxic to tumor cells. However, when cultured with tumor cell fractions sedimented at 1000 X g(R1), 20,000 X g(R2), and 100,000 X g(R3), these lymphocytes became markedly cytotoxic to specific tumor targets in a 3.5-hr (51)Cr release assay. R2 fractions were significantly more immunogenic than were R3 fractions (p less than 0.05). Although lymphocytes sensitized with SH-3 tumor cell fractions were cytotoxic to SH-3 tumor cells, they were also cytotoxic to cells from RPMI-7932 and RPMI-8322 (malignant melanoma) tumor lines and vice versa. Cells from tumor lines HT-29 (colon carcinoma) and COLO 110 (ovary carcinoma) were significantly less susceptible to lysis by effector cells generated against SH-3. These immune cells, although capable of killing cells from tumor lines, were not able to lyse cells from autochthonous normal lymphoid lines or normal lymphocytes that have been transformed by phytohemagglutinin. Tumor cell fractions were not immunogenic at low (5- to 20-mul/0.75 ml) concentrations; an increase of 4- to 10- fold in their concentrations was usually followed by a decrease in immunization.


Treatment of metastatic germ-cell tumors in men with adriamycin, vincristine, and bleomycin. Twenty-five patients with metastatic germ-cell tumors were treated with a combination of adriamycin, vincristine, and bleomycin. Fifteen patients (60%) had previously received radiation therapy, chemotherapy, or both. Twenty patients (80%) responded to treatment, with eight (32%) patients achieving a complete remission (CR) and 12 (48%) patients achieving a partial remission (PR). The median duration of response for the patients with PR was 4 months, whereas four patients with a CR remain alive without evidence of disease for greater than 3 years after the initiation of treatment. Responses were observed in all tumor categories and apparently were not influenced by prior therapy. Side effects included gastrointestinal toxicity, alopecia, neuropathy, skin changes, mucositis, and myelosuppression (more severe in previously treated patients). Though moderate success was demonstrated for this chemotherapy regimen, it does not appear as effective as more recent regimens including vinblastine and bleomycin.


Specificity of antisera produced against bleomycin. The specificity of antisera produced in a goat or rabbit for use in bleomycin radioimmunoassay has been examined using competitive experiments employing several bleomycin analogs. These studies demonstrate that the antiserum distinguished alterations in both the nucleus and the terminal structure of the molecule. Removal of the terminal structure or addition of an aminosugar moiety in the bleomycin nucleus resulted in a significant decrease in immunoreactivity. Antisera produced in goat and rabbit hosts had different specificity for the terminal structure. The goat antisera demonstrated greater specificity for the terminal side chain than the rabbit antisera although this may not reflect a species-related event.


The optic tectum of calamoichthys calabaricus Smithi. A Golgi study. The optic tectum of Calamoichthys calabaricus (Polypteriformes) shows a relatively complex vertical stratification, with six main layers and a varied neuronal typology. In particular, pyriform neurons in the well developed stratum griseum periventriculare and some multipolar neurons in the stratum griseum profundum represent the efferent elements of the tectum, while the optic and lemniscal inputs to the tectum converge in the plexiform sublayers of the stratum fibrosum et griseum superficiale. In the circuitry of the tectum some modulation is achieved by some of the polymorphic cells of the stratum griseum internum and by the horizontal cells of the outer layers. Notwithstanding some differences with respect to the teleost optic lobe (i.e., the absence of a torus longitudinalis; the lack of a stratum fibrosum marginale; the modest size of the stratum fibrosum profundum; the paucity of neurons in the stratum fibrosum et griseum fibrosum profundum; the paucity of neurons in the stratum fibrosum et griseum superficiale; and the ill-defined separation of the layers of the afferent and efferent fibers), the optic tectum of Calamoichthys resembles the mesotectal type chracteristic of teleost, anurans and reptiles. It exhibits higher degree of organization than the optic tectum of the Chondrostei.


Secretory hyperactivity and mitochondrial changes in neurosecretory cells of an insect. Cellular effects of the insecticide lindane. The neurosecretory cells of the corpus cardiacum of the desert locust Schistocerca gregaria secret their hormonal products by exocytosis. The insecticide lindane is known to cause release of hyperglycaemic but adipokinetic neurohormones. Electron microscopy of lindane-poisoned corpora cardiaca reveals many exocytotic omega figures. Mitochondria are affected either directly by the poison or by the consequences of secretory hyperactivity. They occur in increased number, divide, acquire dense mitochondrial granules larger than in the controls, and sometimes line up along plasma membranes, mitochondria in two neighbouring cells forming pairs in juxtaposition. Between two such mitochondria the plasma membranes form a junction-like structure. It is suggested that these effects reflect an excessive calcium entry caused by lindane.


Localization of gonadotrophic hormones in the dog pituitary gland. A study using immunoenzyme histochemistry and chemical staining. Using the immunoperoxidase technique and antisera to the specific beta (beta) subunits of FSH and LH1, selective immunochemical staining was localized mostly in the same cell type in the pars distalis and pars tuberalis of the dog pituitary gland. However, some cells were consistently shown to react solely with antisera to either LH beta of FSH beta. The cells stained for FSH beta were at least 1.5 times less numerous than those shown to contain LH beta. In the pars distalis of adult male dogs the immunoreactive gonadotrophs varied greatly in their relative proportion and were mostly shown to be much less numerous than in bitches in the anestrus phase of the sexual cycle. These cells were found to be positive to aldehyde fuchsin, alcian blue, periodic acid-Schiff (PAS) and aniline blue. The performic acid-alcian blue (pH 0.2)-PAS-orange G procedure stained the FSH/LH cells blue or turquoise, demonstrating TSH cells (blue-purple), ACTH/MSH cells (red-purple) and PRL cells (orange-red). The FSH/LH cells were further differentiated from other functional cell types of the pars distalis  on the basis of their typical cytological features, intraglandular distribution and by immunochemical double staining. These observations support the concept that the one cell-one hormone theory may not apply to gonadotrophic hormones, although some cells seem to be the source of either FSH or LH.


An electron microscopic investigation of the surface coat of the electrocyte of electrophorus electricus. The surface coat of the electrocyte of the main electric organ of Electrophorus electricus was studied using cytochemical methods (periodic acid-silver methanamine, periodic acid-chromic acid-silver methenamine, periodic acid-thiosemicarbazide-silver proteinate, Concanavalin A - horseradish peroxidase, ruthenium red, Alcian-blue lanthanum nitrate, colloidal iron hydroxide and cationized ferritin). The surface of the electrocyte presents perpendicularly oriented tubular invaginations of the cell membrane. The fibrous coat 50-100 nm thick, penetrates into the lumen of the invaginations. It is also observed in the synaptic clefts existent in the posterior face of the electrolyte. The coating of the surface membrane gives a positive reaction with all techniques used. Binding of colloidal iron hydroxide particles was observed only in the outer layer of the coat. With the Alcian-blue lanthanum nitrate technique , microtubules were observed in the cytoplasm of the electrocyte. The results indicate that the surface coat of the electrocyte contains mucopolysaccharides, glycoproteins, acid mucopolysaccharides and anionic sites detected at low (colloidal iron hydroxyde) and neutral (cationized ferritin) pH.


Somatopetal transport of horseradish peroxidase (HRP) in the peripheral and central branches of dorsal root ganglion cells. The axonal transport of HRP in both the peripheral and central branches of dorsal root ganglion cells was studied in rats. For studying axonal transport in the peripheral branch HRP as a dry substance was applied to the peroneal nerve injured either by teasing, by cutting or crushing. After a short survival time (22 h) mainly small spinal ganglion cells of the corresponding segments were labelled, while after a prolonged survival time (70 h) mainly large cells were labelled. These labelling differences are referred to different transport rates or to differences in the process of accumulation of HRP in neurons of various sizes. No evidence could be found for HRP transport from the peripheral into the central branch. Injection of HRP into the spinal cord (survival time 22 h) or into the dorsal column nuclei (survival time 46 h) was followed by labelling of numerous spinal ganglion cell perikarya of all sizes. Reaction product was found also within the prebifurcation segment of spinal ganglion cell processes. On the basis of light microscopic exploration only somatopetal transport could be detected.


Localization of serum-derived alpha 2 macroglobulin in cultured cells and decrease after Moloney sarcoma virus transformation. NRK cells and many other cultured fibroblasts were found to contain the protease inhibitor, alpha 2 macroglobulin (alpha 2M). This alpha 2M is present as a result of uptake of alpha 2M from the calf serum in the culture medium. Some of this alpha 2M is released back into the medium. In radiolabeling experiments with 14C-amino acids, no radioactivity was detected in intracellular or extracellular alpha 2M. Fluorescence microscopy of fixed cells using rhodamine-labeled antibodies indicated that alpha 2M is present in vesicular organelles different from primary lysosomes. Fluorescence microscopy of living cells shows that rhodamine-labeled alpha 2M (rhodamine-alpha 2M) is taken up into similar structures. Of the many cell lines examined, Moloney sarcoma virus-transformed cells had the lowest amounts of alpha 2M. Some of the effects of serum on the behavior of cultured cells could be a consequence of inhibition of cellular proteases by alpha 2M.


Properties of a P70 proteolytic factor of murine leukemia viruses. Murine leukemia viruses, such as Rauscher leukemia virus (RLV), contain a proteolytic factor which becomes activated after detergent treatment of the virus. This factor specifically cleaves P70, the gag precursor polyprotein which is enriched for in preparations of immature virus core subparticles. The factor has been partially purified on Sephadex G-75 columns. It has a molecular weight of 10,000-12,000 daltons but does not coincide in elution position with the major peaks of the viral polypeptides p10 or p12. Under optimal conditions, that is 2% NP-40 (v/v), 10 mM DTT, (pH 7.2) and incubation for 16 hr at 22 degrees C, cleavage of labeled P70 occurs and increasing amounts of the four gag polypeptides p30, p15, p12 and p10 are obtained. The P70 cleavage activity is blocked by TLCK, TAME, CBZ-lysine and other lysyl-containing protease inhibitors. Further, the CBZ-lysine inhibition is reversible, while an inhibition by phenyl-methylsulfonyl fluoride (PMSF) is irreversible. These inhibition studies suggest that a similarity exists between the P70 proteolytic factor and some serine proteases, such as trypsin. The cleavage pattern of P70-rich immature cores treated with trypsin or chymotrypsin is different from that obtained with the P70 proteolytic factor. Thus murine leukemia virions apparently contain a unique, highly specific protease which is present in small amounts and cleaves P70.


[Palliative treatment of liver neoplasms I: localized infusion of antimitotics: premises, technical procedure and limitations]. A hepatic localised antiblastic chemotherapic technique combining limited discomfort for patients with appreciable palliative results is illustrated; a short description of the surgical procedure of intubation of the hepatic artery itself and of some auxiliary procedures is given. Lastly the possible complications and the results are illustrated.


[Palliative treatment of liver neoplasms II: dearterialization of the liver: indications, premises, technic and limitations]. Hepatic arterial vascularisation, as described by Michels in 1954, is outlined; the importance of selective angiography in preparation of the hepatic de-arterialisation operation is then stressed, and its theoretical foundations and the surgical technique are illustrated; other procedures which can usefully be combined are also indicated. Lastly an evaluation of the limits and results of this therapy is given.


Isoelectric focusing of gamma globulins in cerebrospinal fluid from patients with multiple sclerosis. The technique of isoelectric focusing has been adapted for rapid clinical analysis for globulins in cerebrospinal fluid with use of commercially prepared horizontal-slab acrylamide gels. The globulin fraction is concentrated by ammonium sulfate precipitation, which allows more of the relevant protein to be applied, use of a wider range of total protein concentrations, and higher resolution than is true for previously described methods. Critical variables include a constant concentration and volume of IgG, a constant low temperature of the acrylamide gel, and sensitive staining with Coomassie Brilliant Blue G-250. The apparatus used is adaptable for other electrophoretic procedures in the clinical laboratory, and the use of commercially prepared gel slabs is more convenient, more reproducible, and requires less time than other methods.


Acute-phase proteins from the liver and enzymes from myocardial infarction; a quantitative relationship. In 14 patients with acute myocardial infarction (M.I.) not having any other disease, the possible quantitative relationship between enzymes from M.I. and changes in concentration of acute phase reactants coming from the liver were studied. The patients were followed up until 1 1/2 years after M.I. and comparison of baseline-protein values took place using a control group of 18 healthy individuals. Quantitation of protein changes was done by planimetric determination of the area under the concentration curve and by taking peak values. The myocardial infarction was quantitatively estimated by mathematical analysis of the time course of alpha-hydroxybutyrate dehydrogenase (alpha-HBDH) plasma concentrations and by taking peak values. A quantitative relationship with enzymatic infarct size was found for haptoglobin, alpha1-acid glycoprotein, alpha1-antitrypsin, C-reactive protein, fibrinogen and E.S.R. Albumin and transferrin did not show a negative quantitative relationship with enzymatic infarct size. Humoral factors originating from the site of tissue injury and evoking in proportion a positive acute phase reaction by the liver are probably the basis for this observed quantitative relationship.


[Artistic production in epilepsy].

[Artistic production in epilepsy]. In the epileptic's relation to the world, there is--as a particular way of being--an opposition between two constitutive moments: the world of the dark mass and the world of the light tone of colour as moments of an entirety. This can be seen symbolically in pictures as a contrast between light with an upward trend towards heaven and darkness as an earthy mass with a downward tendency. The two worlds are one at the same time, with diverging trends coexisting in a dynamically flowing organic interrelationship. This is combined with a specific religious atmosphere. From this point of view the attempt is made to gain an understanding of the condition of being ill with epilepsy.


Prolonged administration of bleomycin without clinical toxicity. Therapy with 2,700 units over four years. A 26-year-old white man with stage 4-B Hodgkin's disease resistant to conventional chemotherapy obtained complete remission with administration of bleomycin. Maintenance of this remission required continued therapy with bleomycin. The patient received a total of 2,700 units of bleomycin over a 51-month period without signs or symptoms of pulmonary toxicity. Serial studies of pulmonary function have shown a stable forced vital capacity and minimally decreased total lung volume and pulmonary diffusing capacity. This case demonstrates the ability of a patient to tolerate massive cumulative doses ob bleomycin over a protracted period without severe loss of pulmonary function.


Differential Giemsa staining of sister chromatids after extraction with acids. Chromosomes of Chinese hamster strain cells were air-dried on slides after BrdU substitution for two or three rounds of replication. The preparations were treated with 20% PCA at 55 degrees C for 20-30 min, or 5N HCl at 55 degrees C for 15-20 min. After staining with Giemsa, unifilarly BrdU-substituted chromatids stained faintly and bifilarly substituted chromatids stained darkly. Such a pattern of sister chromatid differential staining was confirmed by the examination of metaphase cells grown with BrdU for three rounds of replication.


R-banding produced by DNase I digestion of chromomycin-stained chromosomes. A distinct reverse (R-) banding pattern was produced on human chromosomes by digesting chromosome spreads with pancreatic deoxyribonuclease I (DNase I) in the presence of an excess of chromomycin A3 (CMA), followed by staining with Giemsa. The banding pattern corresponds with that obtained by chromomycin A3 fluorescence, and bands which fluorescence brightly with chromomycin appear darkly with Giemsa. The same relationship was observed in two plants, Scilla siberica and Ornithogalum caudatum, which have contrasting types of heterochromatin. Chromomycin bright C-bands stained darkly with the CMA/DNase I technique, whereas chromomycin negative C-bands appeared lightly stained. The digestion patterns are thought to reflect the variation in chromomycin binding capacity along the chromosome with R-bands and dark C-bands being sites which preferentially bind the antibiotic.


[Acquired agammaglobulinaemia with predominantly intestinal symptoms (author's transl)]. In a 45-year-old female patient primary acquired agammaglobulinaemia was diagnosed. Intestinal symptoms predominated. The disease was characterized by a B-cell defect. Substitution with gamma-globulin (Beriglobin) practically cured the symptoms. The pathogenesis of the disease remains unexplained.


[Immunological changes during hydrotherapy]. Immunological tests were performed in 34 patients undergoing hydrotherapy according to Kneipp. IgM, alpha2-macroglobulin and complement factor C3 concentrations were increased after this treatment, but not in ten healthy volunteers. Specific antibody activities (staphylococci, E. coli, streptococci) remained unchanged. After intracutaneous testing with different protein and bacterial antigens reactions were significantly more intense (diameter of reaction) after hydrotherapy, while total number of reacting patients remained the same. Pokeweed stimulation ratio was increased in the lymphocyte transformation test, while PHA and PPD stimulation rates remained unaltered.


Far field somatosensory evoked potentials in the cat. Averaged somatosensory evoked potentials (SEP) after forepaw stimulation were recorded from skull electrodes in the cat. The cortical component of the SEP was preceded by four far-field potentials (Component I, II, III, IV). A study of these components suggests that: (1) Component I originates in the cervical spinal cord; (2) Component II is generated in the nucleus cuneatus and/or medial lemniscus; (3) Component III-A recorded widely over the skull probably reflects activity in the cerebellum and/or cerebellar pathways; (4) Component III-B seen over the somato-sensory area results from superposition of component III-A and activity in the ventral posterior nucleus of the  thalamus and (5) Component IV orginates in the sensory radiation with some contribution from the thalamus.


Memory retention of stimulations during REM and NREM stages of sleep. In 51 normal young female subjects, stimulation by name calling or by intermittent photic stimulation was given during sleep. At different intervals after the stimulation, the subject was awakened and asked if she could recall it. If alpha activity had not been elicited by the stimulus, there was no recall. If the time occupied by alpha activity evoked by the stimulation was more than 30 sec, the stimulus could be recalled after a long period of sleep. When the evoked alpha activity lasted for less than 30 sec and the subject slept again, the longer the evoked alpha activity, the longer the sleeping time span with the memory retention of the preceding stimulation. With equal durations of evoked alpha activity, retention of the stimulus was better when the sleep following was REM stage than when it was NREM stage (stage 2). The results might be explained by the assumption that process of consolidation takes place most rapidly during wakefulness and is inhibited during sleep but to a lesser degree during REM stage than during NREM stage (stage 2).


Differential effects of cortical speading depression on epileptic foci induced by various convulsants. Interaction between epileptic foci and spreading depression (SD) was studied in the cerebral cortex of rats anesthetized with Nembutal. At comparable discharge rates, picrotoxin and penicillin caused complete and partial SD blockade respectively, strychnine and Metrazol were ineffective and Aldactone facilitated SD. Conversely, the duration of SD-induced blockade of epileptic activity was maximal for Aldactone and minimal for picrotoxin. Treatment of the picrotoxin focus with tetrodotoxin (10(-4)M) reduced the discharge rate and reinstated SD propagation into the focus. [K+]e measured with ion-selective K+ electrodes 1 mm below the cortical surface increased to 8 mM in penicillin foci blocking SD and remained below 5 mM in Aldactone foci. It is concluded that the differential effect of various convulsants on SD propagation depends on the potassium concentration in the depth of the focus rather than on the discharge rate or on the mechanism of the the epileptogenic effect.


Modification of amygdalo-cortical evoked potentials by kindling and pentetrazol-induced generalized convulsions in cats. Amygdalo-cortical pathways were studied in control cats, kindled cats and pentetrazol-injected cats by the evoked potential method. An increase in amplitude of suprasylvian cortical evoked potentials to stimulation of the amygdala was observed in the kindled group and in the pentetrazol group. Moreover, evoked potentials to stimulation of the amygdala appeared at the sigmoid gyrus after generalized convulsions in the kindled group and after pentetrazol in the other group. These modifications were permanent in the kindled group while they were only transient in the pentetrazol group.


Sleep studies on canine narcolepsy: pattern and cycle comparisons between affected and normal dogs. Two narcoleptic and 2 normal poodle or mixed poodle dogs were polygraphically monitored for 48 h with one narcoleptic and one normal monitored concurrently. Data were categorized by 15-sec epochs into wakefulness, light sleep, slow wave sleep, REM sleep, cataplexy (immobility preceded by wakefulness with partial or complete electromyographic quiescence and pronounced theta activity from subcortical leads), and atonia with no theta (15-30 sec periods like cataplexy but without theta). In narcoleptics we could see no gross differences between the polygraphic records of cataplexy and those of REM sleep; scoring distinctions between the two states depended on the antecedent state. Results indicated that narcoleptic dogs do not differ from normals with respect to percent of time spent in wakefulness (39.8% vs. 42.6%), light sleep (16.2% vs. 18.4%), or slow wave sleep (27.2% vs. 28.0%). Narcoleptic dogs spent slightly less time than normals in REM sleep (6.9% vs. 11.1%) and spend 9.1% and 0.8% of the recording time in cataplexy and atonia with no theta respectively. Normal dogs presented neither of these pathological states.


Effects of benzodiazepines on PGO firings and multiple unit activity in the midbrain reticular formation in cats. Effects of benzodiapines administered by the intraperitoneal route on PGO firings and multiple unit activity in the midbrain reticular formation in chronic cat preparations were investigated at various levels of consciousness. Changes in the sleep-wakefulness cycle induced by direct injection of benzodiazepines into the reticular formation were also investigated. Benzodiazepines markedly decreased multiple unit activity in the midbrain reticular formation during each stage of sleep, but had little effect during behavioral and EEG arousal. Benzodiazepines did not affect PGO firing rate, but attenuated all increase of multiple unit activity following PGO firings. The bilateral injection of benzodiazepines into the midbrain reticular formation induced an increase of arousal and a decrease of slow wave sleep, but did not change the amount of paradoxical sleep. It is concluded that benzodiazepines show a mixture of depressant and facilitatory effects, which seem to vary with the state of consciousness of the animal.


Average multichannel EEG potential fields evoked from upper and lower hemi-retina: latency differences. Checkerboard reversals shown to the upper hemiretina evoke EEG potentials which in anterior-posterior derivations are approximately inverted in polarity compared with potentials evoked by lower hemiretinal stimulation. Using a 48-channel system, EEG scalp field distributions of such responses were mapped. Examination of the map sequences shows that occipital positive maximal field values start to develop at about the same time in the two stimulus conditions, but peak much earlier and somewhat more anteriorly for upper than for lower hemiretinal stimulation. Thus, there is a difference of response latency in the two conditions, which accounts for the approximate inversion of polarity. Possible correlations with reports of higher receptor cell density, higher visual acuity, and shorter motor reaction time of the upper hemiretinal system are noted.


[Alterations of sleep caused by acute and chronic adminstration of lithium]. The sleep of 2 groups of subjects receiving lithium was studied. Ten normal controls after 17 days of lithium had a significant decrease of Paradoxical Sleep duration, with an increase of PS latency. A second group of 10 manic-depressive patients had received lithium for 22 +/- 6 months when they were recorded. They had a greater amount of slow wave sleep than the control group before lithium but the difference of PS duration was only related to a first night effect. Therefore sleep changes induced by lithium are different according to the duration of lithium administration. In any case they are less important than those induced by most of the anti-depressants.


A proposed methodology for evaluation of the Wiener filtering method of evoked potential estimation. A method for comparing signal averaging and a posteriori Weiner filtering as noise reduction methods for evoked potential estimation is proposed.


Development of the single microelectrode current and voltage clamp for central nervous system neurons. The development of a single microelectrode clamping circuit for central nervous system preparations is presented. The efficacy of the circuit in both current and voltage clamp modes is demonstrated by investigating the membrane properties of cat spinal motoneurons. Results are compared with those obtained using conventional techniques and reveal that this technique can be more readily applied to central nervous system neurons. The constraints invovled in employing this system to study time-varying events are also discussed.


Graphical display and statistical evaluation of event-related desynchronization (ERD). A new method which enables quantification of event-related desynchronization (ERD) by measuring changes in the power of rhythmic activity within the alpha band during sensory stimulation and during the interstimulus interval of a CNV paradigm is described. For the processing, EEG trials recorded under the same stimulus conditions and with eyes closed are used. For the power changes, the statistical significance is tested with a non-parametric test. With the proposed method it is possible to measure very brief power changes of the order of 21 percent (corresponding to amplitude changes of 11%). An ERD may thus be observed in a restricted cortical region.


Serum thyrotropin response to thyrotropin-releasing hormone and free thyroid hormone indices in patients with familial thyroxine-binding globulin deficiency. The response in serum thyrotropin (TSH) to synthetic thyrotropin-releasing hormone (TRH) as well as serum free thyroxine index (FT4I) and free triiodothyronine index (FT3I) was investigated in six patients with familial thyroxine-binding-globulin (TBG) deficiency. The total serum thyroxine (T4) and triiodothyronine (T3) concentrations were significantly decreased, compared with those of normal subjects (3.4 +/- 0.9 microgram/dl, mean +/- SD. vs. 9.0 +/- 1.5 microgram/dl, p less than 0.01 and 87 +/- 27 ng/dl vs. 153 +/- 37 ng/dl, p less than 0.01, respectively). FT4I was lower than the normal range in all but one (5.3 +/- 1.5 vs. 8.9 +/- 1.6, p less than 0.01), whereas FT3I was all in the normal range and of no significant difference from the normal control (132 +/- 22 vs. 148 +/- 25). Serum TSH concentrations in TBG deficiency were all in the normal range (1.0-4.2 muU/ml) and the maximum TSH increments following TRH 500 microgram iv were 8.9 +/- 2.0 muU/ml and of no significant difference from the normal control (10.2 +/- 4.5 muU/ml). These results indicate that the euthyroid state in familial TBG deficiency is more clearly defined by TRH-test and the normal response to TRH in familial TBG deficiency is presumably under the control of the serum free T3 level rather than the serum free T4 level.


Effects of total fasting in obese women. I. Response of the thyroid. The values of serum T4, T3, T4/T3, RT3U, FT4I, PBI, ATR, TSH, 132I uptake and antibodies against thyreoglobulin were estimated in non-obese, healthy women and in obese women with a stabile body weight on total fasting of 14 days. Obesity with no dietary restriction compared with normal controls was accompanied by a tendency to higher serum levels of T4 and T3 and prolonged ATR. Other indicators of thyroid function remained unchanged. Total fasting led to a decrease of serum T3 and to an increase of FT4I and RT3U. The remaining tests were unchanged. These results are conform with a decreased transformation of T4 into T3 in the periphery. The participation of the hypothalamus and hypophysis to adaptation of the thyroid to fasting was discussed.


Kinetics of refolding of completely reduced human-serum albumin. Regain of immunochemical reactivity. The kinetics of refolding of completely reduced human serum albumin has been studied by various methods including immunological techniques. The decrease in thiol content is very rapid in the beginning of the reoxidation process and rather slow in the later stages. Polyacrylamide gel electrophoresis studies show that, in the earlier stages of refolding, the main part of the albumin is present as various oligomers and that a slow conversion to monomer occurs as reoxidation proceeds. Rocket immunoelectrophoresis shows that the completely reduced protein is devoid of native albumin antigenic determinants but that a rapid regain of immunoprecipitability is obtained upon reoxidation. A new 'consumption' rocket immunoelectrophoretic method has been used to estimate the total regain of antigenicity. The data obtained indicate that there is a preferential rapid folding to native structure in certain parts of the molecule but that areas with wrong or incomplete foldings exist a considerable time after the inital refolding period.


Purification and further characterization of an RNA-dependent DNA polymerase from the allantoic fluid of leukosis-virus-free chicken eggs. The purification of an RNA-dependent DNA polymerase from the allantoic fluid of uninfected, embryonated chicken eggs is described in detail. Comparison to the polymerase of avian myeloblastosis virus shows that the two enzymes are different with respect to ion concentrations for optimal reaction, response to increasing concentrations of substrate, thermal stability and protection from thermal inactivation by viral RNA. It is concluded that the enzymes compared to each other are different proteins, which must have been coded by different genes. The RNA-dependent DNA polymerase in the allantoic fluid, therefore, does not derive from the partial or complete expression of the endogenous virus genome of the normal chicken cell or from infection by exogenous viruses.


Biochemical markers of the progress of differentiation in cloned teratocarcinoma cell lines. The progeny of single teratocarcinoma cells will give rise to several different cell types in vitro, and the latter were shown to be functionally differentiated by biochemical criteria. In all these studies, cloned lines of mouse teratocarcinoma cells were assayed during the course of differentiation for some biochemical products characteristic of the tissues formed. The carcinoembryonic protein, alpha-foetoprotein, was not synthesized by undifferentiated embryonal carcinoma (EC) cells, but was synthesized in increasing amounts during their differentiation to endoderm-type cells in suspension culture. alpha-Foetoprotein was shown to be a product of endoderm cells, but not all endoderm cells synthesized this protein. During the course of further differentiation when EC cells or aggregates were grown in tissue-culture dishes, other biochemical products appeared. In cultures containing predominantly nerve-type cells, there was a 30-fold increase in the specific activity of acetylcholinesterase, with concomitant appearance of the aldolase isoenzyme characteristic of mouse brain. In some cultures, a small amount of muscle-type cell formation was marked by the appearance of the MB isoenzyme of creatine phosphokinase. Generally, biochemical differentiation was immature.


57-cobalt-bleomycin as a tumor scanning agent in primary hepatocellular cancer. 57Co-bleomycin and 99mTc-sulfur colloid scans were performed in 13 southern African black patients with primary hepatocellular cancer (PHC) and 3 patients with other space-occupying hepatic lesions. Selective concentration of 57Co-bleomycin in the defect or defects seen on the 99mTc-sulfur colloid scan occurred in only 4 of the 13 (31%) patients with PHC. In the remaining PHC patients, as well as in the 2 patients with metastatic liver disease and the 1 patient with an amebic liver abscess, the 57 Co-bleomycin and 99mTc-sulfur colloid scans were identical. 57Co-bleomycin appears to have only a limited place as a tumor-seeking agent in the diagnosis of PHC in southern African blacks.


The possible role of vasoactive substances for the hemodynamic effects of rapidly injected antilymphocyte globulin. A rapid intravenous injection of ALG into splenectomized dogs induced an almost complete elimination of lymphocytes and an extensive but partly reversible thrombocytopenia during which platelets were trapped in the lung. During this trapping there was a significant decrease in arterial blood pressure, pulmonary blood volume and cardiac output concomitant with a marked increase in the portal pressure. The pulmonary arterial pressure remained unchanged. The hemodynamic effects are probably caused by a release of vasoactive substances which induced a decreased venous return from splanchnic pooling and statis. Prostaglandins and histamine seem to play a dominant role, since Aspirin, promethazine and 48/80 markedly reduced the side-effects.


Human B cell line deficient in the expression of B cell-specific glycoproteins (GP 27,35). A human B lymphoid cell line, P3HR-1, expresses only low levels of the 27 000 and 35 000 mol.wt. B cell-specific glycoproteins (GP 27,35). Indirect antibody-binding and quantitative absorption tests with a xenoantiserum against the antigens showed that P3HR-1 cells have on their surface about 1% of the amount found on other human B lymphoblastoid cell lines. The deficit of the glycoproteins on the surface of P3HR-1 cells could be accounted for by a reduced rate of synthesis in these cells. A simple relationship between the reduced expression of GP 27,35 on P3HR-1 cells and their inability to bind Epstein-Barr virus (EBV) or express complement receptors was excluded because other B lymphoid cells which expressed neither virus-binding sites nor complement receptor had normal amounts of GP 27,35 on their surface. However, antibodies against GP 27,35 could block the absorption of EBV by EBV receptor-positive B cells.


Role of epitope density in the induction of immunity and tolerance with thymus-independent antigens. IV. Selective tolerance and IgE response by DNP-levan conjugates. Tolerance induction of IgE antibody-forming cells by dinitrophenylated levan (DNP-LE) conjugates was investigated in CBA mice. Antibody production was measured by passive coutaneous anaphylaxis on rat skin. Specific antibody neutralization was obtained with conjugates of different substitution degrees. Specific inhibition of IgE antibody production was obtained only with highly substituted conjugates. However, the epitope density required for inducing tolerance to antibody synthesis of the IgE class was lower than for other classes of immunoglobulins. The tolerance induced by DNP-LE was shown to affect only the anti-DNP B cells of IgE class without T cell participation. These results demonstrate that B cell precursors of all Ig classes are susceptible to tolerance induction and not to triggering by T-independent antigens.


Characterization of syngeneic anti-idiotypic antibody against the idiotype fo BALB/c myeloma protein J558. BALB/c mice were immunized against the idiotype of BALB/c myeloma J55,. Syngeneic, specific anti-idiotypic antibodies against this germ line idiotype were shown by the isoelectric focusing technique to be markedly heterogeneous. In fact, the heterogeneity of isogeneic anti-idiotypic sera appeared to be comparable to those of allogeneic anti-J558 sera, raised in A/J and CB20 mice. As a rule, individual mice exhibit different clonal spectra. By spleen cell transfer experiments, each individual spectrum of clones could be expanded in order to arrive at an estimate of the size of anti-idiotypic repertoires. These were found to be of the order of 7-16 different anti-idiotypic clones for an individual BALB/c mouse. From the infrequency of clonal repetition we conclude that the repertoire of the BALB/c strain must be well in excess of one hundred anti-idiotypic molecular species.


Antigenic correlation between rat thymus and superior cervical ganglion. By using rabbit anti-rat thymocyte and anti-rat superior cervical ganglion sera in cytotoxicity, immunofluorescence and absorption assays, it has been shown that surface membranes of rat thymocytes and cervical ganglion cells (i.e. peripheral nervous tissue cells) contain common antigenic determinants.


Gastrin: obligatory intermediate in the postprandial mobilization of gastric histamine in the rat. In unoperated fasted rats, feeding raised the serum gastrin concentration, reduced the gastric mucosal histamine content and activated the gastric histidine decarboxylase. The reduction of gastric histamine and activation of histidine decarboxylase was induced also by the injection of pentagastrin. In antrectomized rats, feeding failed to produce these effects. Injection of pentagastrin, however, still lowered gastric histamine and activated gastric histidine decarboxylase. Thus, antral gastrin seems to be an obligatory mediator of the postprandial activation of histidine decarboxylase and mobilization of histamine.


[Comparative characteristics of electrophysiologic changes in frog muscle fibers after denervation and following blockade of axoplasmic transport]. Using anatomical denervation and a single application of colchicine the k ACh sensitivity and electric features of the membrane of the m. sartorius fibers were studied. Within 2 weeks after anatomical denervation a similar enlargement of the area sensitive to ACh occurred as well as a decrease in the MP values, in effective impedance, and in time constant of the muscular membrane. These data suggest that  the postdenervation changes in the frog muscle are due mainly to cessation of the substances inflow with the fast axoplasmatic transport. Two latter parameters change at that more rapidly after colchicin which suggests a greater effect of the deficit of trophic substances transported along the axons, on the functioning muscle (colchicine) than on the inactive (denervation).


[Study of the histaminergic mechanisms of the action of malaben]. In rabbits (intact and with experimental myocardial infarction) histamine metabolism (histamine content and diaminoxidase activity) following introduction of malaben was studied. In intact animals the ability of malaben to reduce the blood histamine level and to activate diaminoxidase was discovered. Administration of malaben in experimental myocardial infarction promotes a quicker normalization of the disturbed metabolism of histamine.


Alternatives to female sterilization.

Alternatives to female sterilization. With national policies aimed at reaching zero population growth in the shortest possible time, many countries have introduced restrictive legislation and disincentive programs in an attempt to decrease family size norms to 2 or 3 children. As a result, younger women of lower parity are being sterilized, with consequences that will be seen in the years to come. Although female sterilization is usually associated with minimal complications and side effects and is highly effective without continuing motivation or promotion, it has the disadvantage of causing permanent, essentially irreversible sterility. Therefore, it will not be readily acceptable to a large portion of the population. For many women, alternatives must be sought. Reversible methods of female or male sterilization, longacting systemic contraceptives, longacting implants, and immunization against implantation or sperm antigens are potential alternatives, but all are still in experimental stages of development. Intrauterine devices and injectables are the 2 most effective alternatives now available. The use of intrauterine devices with abortion as a backup in case of contraceptive failure ranks high as an alternative to female sterilization.


Preparation and assessment of antisera to ACTH. Antisera to ACTH were produced in rabbits injected repeatedly at multiple intradermal sites with synthetic [Asp25, Ala26, Gly27]alphah-corticotropin-(1-28)-octacosapeptide-bovine gamma globulin conjugate (octacosapeptide is a sequence analogue of alphah1-28-ACTH). Antibodies to extracted human or porcine ACTH were detected in all of the sera 1 month after immunization. A considerable proportion of the antisera obtained from a single final bleeding 5 months after the primary immunization were suitable for sensitive radioimmunoassay. The antisera were shown to neutralize the steroidogenic activity of ACTH in an isolated rat adrenal cell bioassay system. Titres estimated from antiserum dilution curves and relative avidities from the standard curves were compared. It was possible to detect picogram amounts of ACTH in plasma-free medium with the best antisera. The method described is an effective means of producing anti-sera to the weakly immunogenic N-terminal fragment of the ACTH molecule.


Primary sarcoma of the liver. Report of a case with excellent response to hepatic artery ligation and infusion chemotherapy. A case of primary undifferentiated sarcoma of the liver in a 66-year-old woman is reported. The similarity of the presenting symptoms to those of a hepatic abscess is emphasized. The patient's response to palliative treatment by means of hepatic artery ligation and chemotherapy was excellent. We believe that this is the first  report of such a response in a sarcoma localized to the liver. This therapy may well be useful in symptomatic patients in whom partial hepatectomy is not feasible.


Frameshifts and frameshift suppressors in Saccharomyces cerevisiae. Using ICR-170 as a mutagen, we have induced a set of mutations in yeast which exhibit behavior similar to that shown for bacterial frameshift mutations. Our genetic study shows that these mutations are polar; the polarity can be relieved by internal suppressors; they revert with acridine half-mustards and are not suppressed by known nonsense suppressors. However, they are suppressed by other dominant external suppressors, which fall into two mutually exclusive groups. Five genetically distinct suppressors were obtained for one of these groups, using co-reversion of two frameshift markers. Three of these are lethal in combination with each other and show a reduction in the GLY3 tRNA peak on a Sepharose 4B column. A fourth suppressor shows an altered chromatographic profile for GLY1 tRNA. We suggest that this group of suppressors represent mutations in the structural genes for the isoaccepting glycyl-tRNA's. Two other suppressors (one linked to the centromere of chromosome III) were found to suppress a second group of frameshifts. Genetic and biochemical studies show that the nonMendelian factor (PSI+) increases the efficiency of some frameshift suppressors.


[The number and kind of leukocytes in the saliva (salivary corpuscles) in healthy subjects and in patients]. In the form of a survey a report is given on the concept of the saliva corpuscle, its origin, composition, different degree of damage, its number and function. The quantitative behaviour enables a statement to be made about the actual unspecific cellular defence in parodontopathy (increased number of saliva corpuscles) and in various blood diseases (decreased number of saliva corpuscles. This has a particular diagnostic value.


[The pathogenesis of chronic lymphocytic leukemia. II]. Starting from a survey of the present knowledge of morphological, immunological and biochemical characteristics of lymphocytes degenerated in leukamic respect and their kinetics, proposals are made for the pathogenesis of chronic lymphatic leukaemia (CLL). The fact that a progressing restriction of the functional immunological variety will occur in the typical CLL is discussed. Besides normal T and B cells there are elements similar to B and T cells whose percentage of proportion changes in the course of the illness with diminution of function and increasing immunological defect.


On the presence of lysozyme in the Nuclei of Leukocytes. The purified chromatin of leukocyte nuclei from two patients, one with chronic granulocytic and another with acute myelomonocytic leukemia, has been investigated for lysozyme activity. The chromatin contained 4.8% resp. 4% of the total amount of lysozyme found in the leukocytes. The function of lysozyme in the nucleus remains unclear.


Karyotype 45, XX, --11,2q+ of bone marrow cells in a case of di Guglielmo syndrome. The case of 50-years-old woman with the chronic type of erythraemia (di Guglielmo syndrome) and the karyotype 45, XX, --11,2q+ of bone marrow cells is described. By means of G-banding the karyotype 45,XX, -2, -11, + der (2), t(2;11) (2pter leads to 2qter:: 11q12 leads to 11qter) was established. The karyotype of bone marrow was thus partically monosomic for chromosome No. 11, for its segment 11 (pter leads to q11).


[Studies on the morphology of blood of the European eel (Anguilla agnuilla). III. Studies on monocytic and lymphoid cells]. Investigations concerning the monocytic and lymphocytic cell series were carried out on the peripheral blood and dabbings of the kidney, the spleen and intestines of healthy eels (anguilla anguilla), and eels taken ill with vibrio anguillarum. By means of histological, cytochemical, phaseoptical, and electronmicroscopical examinations independent development lines of the monocytes and lymphocytes were established. The place of origination for the monocytes was found to be the kidney, for the lymphocytes the spleen and the intestines additionally. The evidence of the cytochemical assay of enzymes for the differentiation of these blood cells and their cellular systems is primarily dependent on the distribution of the activity of the unspecific esterase, the naphthol-AS-D-chloroacetate-esterase, the peroxidase and the alkalic phosphatase. Macrophages were - on the basis of enzymocytochemical results - seen as derivatives of the monocytes. Plasma cells and lymphoid cells are transformation forms of the lymphocytes. Our results emphasize the significance especially of cytochemistry for the hematological diagnostics of fish diseases.


The inhibitory effect of methyl palmitate on heteroagglutinin formation in rats. In Wistar rats the immunosuppressive effect of methyl palmitate on the formation of heteroagglutinins against human O-group erythrocytes was followed. An i.v. injection of methyl palmitate both delayed the heteroagglutinin formation and decreased its intensity. The inhibitory effect of methyl palmitate was not accompained by the subsequent hyperactive phase as could be observed in the previous experiments using ethyl palmitate.


Smooth muscle autoantibodies in normal rabbit sera. Cross-reaction with human platelets as demonstrated by immunofluorescence. 27 normal rabbit sera were tested by indirect immunofluorescent technique. All sera stained smooth and skeletal muscle of human and mammalian tissues and human washed platelets. Two sera had ANA activity. These autoantibodies had low titres. Lysed but not intact platelets neutralized the rabbit SMA sera activity.


[The vitamin B 12 serum level at various ages]. A report is presented on the determination of the vitamin B12 content by means of radioassay in the serum of 402 healthy persons aged between 14 to 98 years. With increasing age a significant decrease of the vitamin B12 content could be observed in both sexes after the sixtieth year of age. In the age group between 21 to 30 only the low values of women made an exception, which may be due to the administration of contraceptives.


[On the causes of intra- and post-operative consumption coagulopathies and postoperative thromboembolism]. The determination of the onset of a disseminated intravascular coagulation (DIC) is examined shortly after its intraoperative andpost-operative dissolution with the help of easily performable haematological and physiological clotting tests in 20 patients. In this connection the operation is appreciated as a model even for other processes defined at the beginning, where DIC can be observed. Whereas the aethanol test, the determination of fibrinogen split products (FSP) and the euglobulin lysis time indicate the beginning of DIC more clearly in the form of average values, the aethanol test, partial thromboplastin time and thrombin time have a prognostic value for each patient. As it is too time consuming to determine FSP, the counting of basophilie granulocytes may be used for the diagnosis. In the initial phase of and post-operative DIC will determine the essential share of predisposition to post-operative thromboembolism.


Feulgen type staining of mammalian tissues with Dahlia-SO2. The use of the basic dye, Dahlia, which belongs to triphenylmethane group but without a primary amino group in its molecule has been described as useful in the staining of aldehyde groups of acid hydrolysed DNA in tissue sections following the conventional Feulgen procedure. Dahlia-SO2 prepared with sodium hydrosulphite is highly suitable when used at pH 4-0 to 5-0. The absorption characteristics of the stained nuclei indicate on the peak of maximum absorption at 560 nm, whereas, that of the aqueous dye solution is at 590 nm.


Histochemical observations on the neuroendocrine complex of Heteroptera, II--The endocrine glands. A histochemical analysis of the retrocerebral endocrine glands of four heteropteran bugs, viz. Sphderodema rusticum Fabr., Ranatra filiformis Fabr., Enithares indica Fabr. and Spilostethus pandurus has been made. Histochemical tests for various types of carbohydrates, proteins, lipids, nucleic acids, etc. have been made. The reaction of different types of cells of the endocrine glands viz. chromophil, chromophobe and CA cells have been studied.


[Effect of surugatoxin on celiac ganglia in cats (author's transl)]. Pharmacological properties of surugatoxin (SGTX), the toxic principle from Japanese ivory mollusc (Babylonia japonica), have been studied. In the present paper, the effect of SGTX on celiac ganglia in cats was investigated by measuring blood pressure from femoral artery and regional blood flow in liver (LBF) and spleen (SBF) of anaesthetized cats. SGTX(50 nmol/kg i.v.) markedly inhibited both the hypertension and changes of LBF and SBF in response to splanchnic (preganglionic) nerve stimulation. On the other hand, it did not inhibit the changes of SBF by splenic(postganglionic) nerve stimulation. The hypertension and changes in LBF and SBF in response to DMPP(94.2 nmol/kg i.v.) were abolished by SGTX(50 nmol/kg i.v.), whereas those to McN-A-343 (94.8 nmol/kg i.v.) were not. SGTX usually enhanced the pressor response to McN-A-343. These results indicate that SGTX has a blocking effect on the nicotinic receptor in celiac ganglia as well as in superior cervical ganglia of cats and rats.


Myocardial infarct size: clinicopathologic agreement and discordance. An accurate postmortem method of planimetrically estimating the extent of myocardial infarction was employed in 16 cases. Delineation of necrotic myocardium was enhanced by a macroscopic staining technique, which utilizes a tetrazolium dye. Comparison of infarct size with peak serum creatine phosphokinase levels showed a general correlation between the two that was not statistically significant. Two markedly disparate cases serve to emphasize the need for clinical awareness of the temporal relationship between myocardial infarction and creatinine phosphokinase analysis as well as the possibility of other anatomic sources of elevation of serum enzyme levels. Comparison of infarct sizes in cardiogenic shock and nonshock patients confirms the existence of a significant relationship between a larger myocardial infarct and shock. However, the data from several patients in the group again emphasize the possibility of maintaining a reasonable blood pressure in the face of a massive myocardial infarction or, more importantly, of manifesting "cardiogenic" shock when only a small amount of left ventricular damage has been sustained. The latter possibility may be related to other anatomic events, e.g., bowel infarction, hemorrhage, or possibly right ventricular ischemia, infarction, or dysfunction.


LBA technique in the detection of chromosome variants. I. Chromosomes with known sites of Q variants. A method is described (LBA method) which used DNA replication pattern in the detection of chromosome variants in man. In Part I, results on chromosomes with known sites of Q-variants, i.e., 5 pairs of acrocentrics, as well as 3 and 4, were presented. Fourty-one variants were detected in a total of 40 acrocentrics. Twenty-eight of them were detected only by the LBA technique; 11 of them being in short arms and 17 of them in centromeres. Seven variants, including 4 of those in satellites, were detected only in QFQ-stained metaphases. Six short-arm variants were observed by both methods. It appears that a sequential QFQ-LBA technique is very useful in the detection of variants in D- or G-group chromosomes.


Antibody response to myelin basic protein: comparisons between Lewis rats with experimental allergic encephalomyelitis, and tolerant rats. The hemagglutinating antibody responses of Lewis  rats with experimental allergic encephalomyelitis (EAE) and of rats rendered unresponsive to this autoimmune disease by pretreatment with myelin basic protein (BP) were compared. Most tolerant animals produced low levels of hemagglutinating antibody. Similarly, most rats with EAE also produced anti-BP antibodies. We were unable to correlate hemagglutinin production or titer with protection against disease. Hemagglutination inhibition (HAI) studies reveal cross-reactivity between rat, guinea pig and bovine BP. HAI studies with BP-derived peptides suggest that at least three distinct antibody-binding determinants exist in the BP molecule, and that individual inbred Lewis rats respond differently with respect to antibody production to these sites.


Release of platelet-activating factor and histamine. I. Effect of immune complexes, complement and neutrophils on human and rabbit mastocytes and basophils. Immune complexes (ICs) triggered the degranulation of basophils/mastocytes in rabbit and man, thus releasing histamine and platelet-activating-factor. ICs acted upon basophils/mastocytes through complement activation and generation of anaphylatoxins and through release of cationic proteins from polymorphonuclear neutrophils. Anaphylatoxins and cationic proteins were active on human and rabbit mastocytes and on human basophils but not on rabbit basophils. Therefore, a cascade of cell to cell interactions may be envisaged implicating successively neutrophils, basophils and/or mastocytes and platelets. These interactions result in enhanced vascular permeability that allows ICs to be trapped along filtering basement membranes. This sequence could perpetuate the deposition of ICs, initially triggered by degranulation of basophils and mastocytes through the IgE-dependent mechanism, and furnish us with an explanation for the rather poorly understood involvement of platelets in immunopathology.


Outer-membrane protein and lipopolysaccharide serotyping of Neisseria meningitidis by inhibition of a solid-phase radioimmunoassay. A new procedure involving inhibition of a solid-phase radioimmunoassay was developed for specific determination of the outer-membrane protein and the lipopolysaccharide (LPS) serotypes of meningococci. Antigen was allowed to bind to the wells of a polyvinyl microtiter plate and then reacted with a limiting amount of homologous antibody which had been preincubated with buffer or a standard concentration of inhibiting antigen. The amount of antibody bound per well was quantitated by incubation with excess 125I-labeled goat anti-rabbit immunoglobulin. Typing sera for detecting eight LPS antigens and 18 protein antigens were made in rabbits by use of both the group C and group B bactericidal serotyping strains. Reactions between unabsorbed sera and purified LPS were inhibited in the LPS typing system, whereas reactions between absorbed sera and outer-membrane complex were inhibited in the protein typing system. Outer-membrane complex was used as the inhibiting antigen in both cases. Approximately 97% of the 80 group B and C strains tested were LPS typable, and 80% were protein typable. Of 51 group A strains tested, however, only 22% were LPS typable and 14% were protein typable. Several nonreciprocal correlations between the occurrence of particular LPS and protein serotype antigens on the same strain were observed, but in general the protein and LPS serotype antigens appeared to occur independently.


Characterization of immunoglobulins in Hodgkin cells. Fifteen of 42 formalin-fixed lymph-node biopsies with Hodgkin lesions of different histological types and stages gave positive staining for immuno-globulins by the peroxidase/antiperoxidase method. The immunoglobulins in all the positively stained Hodgkin cells were of IgG type, but staining for IgA, D and E was negative. One of the 15 biopsies was positive for IgM. Thirteen of the 15 positive biopsies were positive for both kappa and lambda. Sequential double staining using two different substrates for peroxidase showed that most of the Hodgkin cells simultaneously contained light chains of both kappa and lambda type. It is concluded that Hodgkin cells do not contain a "monoclonal" immunoglobulin product.