Sucralfate: antipeptic, antiulcer activities and antagonism of gastric emptying. The antiulcerogenic and antipeptic activities of sucralfate (Ulcerlmin), a basic aluminum sucrose sulfate complex, were investigated. In rats, sucralfate inhibited the formation of ulcers induced by pyloric ligation, indometacin and cysteamine. In doses antagonizing forestomach ulcer formation, sucralfate increased the pH of the gastric juice in a dose-related manner. In vitro, at pH 1.9, sucralfate inhibited rat, dog and hog pepsin in a concentration-related manner and also the peptic activity in human gastric juice. Sucralfate may act as an inhibitor of pepsin by precipitating the enzyme or by binding with it reversibly. The antipeptic activity appears to be directly related to the amount of sucralfate in suspension rather than that in solution. In the gastrointestinal tract, the basic nature of sucralfate may enhance the antipeptic activity of the sucralfate molecule. In rats, sucralfate decreases the rate of gastric emptying.


Ontogenic drug studies in calves. II. Changes in salicylate levels and metabolism in calves with diarrhoea. Acetylsalicylic acid (ASA) was administered by oral route to calves and mice. A comparison of plasma levels of salicylates and salicyluric acid was performed in healthy and diarrhoic calves. The calves were infected with E. coli enterotoxin producing strains. During the 6 h observation period increased levels of salicylates were found in all age groups of calves (1-60 days). There were no significant differences in salicyluric acid plasma levels between controls and diarrhoic animals. Intravenous injection of cholera toxin in mice caused lower levels of total salicylates, but increased levels of salicylic acid and salicyluric acid. The importance of adequate animal model is discussed.


Levamisole treatment in ulcus cruris. A double-blind placebo-controlled study. 59 outpatients (27-82 years) with a chronic leg ulcer participated in a 20-week double-blind placebo-controlled trial of levamisole (levamisole 30; placebo 29). They were instructed to take a body weight adjusted number of tablets (2-5) for two consecutive days every week. Previously used topical treatment was kept unchanged. Control examinations were done every two weeks. Double-blind treatment was stopped because of evident failure after a median duration of 8 weeks in 2 levamisole and 8 placebo patients. The percentage of cured patients became significantly superior in the levamisole group from week 8 onwards, so that, by the end of the study, all levamisole patients were cured against only 76% of the controls. Three levamisole patients reported moderate gastric complaints.


Endotoxin, fever and anomalies of development in rabbits. Short communication. E. coli endotoxin was injected i.v. in rabbits (White New Zealands) on day 9 of gestation. The rise of temperatures was examined by thermoelectric recording. On day 29 of pregnancy the rabbits were killed, the uteri examined for resorptions and the fetuses inspected for malformations. The results were compared with those of control animals. The group treated with E. coli endotoxin showed a significant higher resorption and malformation rate. On the basis of the slight increase of temperatures (up to 1.1 degrees C above normal value), it is supposed that the endotoxin-induced fever did not participate in the teratogenic effects observed.


Steady-state plasma levels of flunarizine in chronically treated patients. Plasma levels were measured at different times during long-term flunarizine treatment (duration 1--45 months, median: 6 months) of 50 patients with intermittent claudication. The dose was gradually reduced, within one month, from an initial dose of 30 mg daily to a maintenance dose of 10 mg daily. Plasma levels, measured by HPLC-method, showed considerable individual differences, ranging between less than 0.02 microgram/ml and 0.25 microgram/ml. Levels between 0.05 and 0.1 microgram/ml were found in 46% of the patients. Individual measurements clearly showed dose-related plasma-levels. There were no signs of accumulation. Side-effects were related to the dose rather than to the levels. There was no correlation between clinical improvement and plasma levels. An extensive first-pass metabolism in the liver is suggested as a possible explanation for the varying inter-individual plasma levels.


The absorption of benzoyl peroxide from leg ulcers. Absorption and clinical efficacy of a preparation containing 20% benzoyl peroxide were studied in 20 patients with leg ulcers of varying etiology. Benzoic acid formed from benzoyl peroxide on penetration through the skin cannot be demonstrated in the plasma until three days after commencing application at the earliest. This indicates a building up of deposits of benzoyl peroxide in the skin. The plasma benzoic acid levels in all patients amounted to less than 5 mumol/l and is therefore toxicologically inoffensive. Clinically, the known therapeutic effects of benzoyl peroxide were confirmed. Neither sensitization nor allergic reactions were found in the 20 patients taking part in this study.


I.c.p. increases with 50% nitrous oxide in oxygen in severe head injuries during controlled ventilation. In a randomized trial nitrous oxide 50% in oxygen (Entonox) or oxygen 100% was given during chest physiotherapy on 23 occasions to three mechanically ventilated patients with severe head injuries. Intracranial pressure (i.c.p.) increased by 22.7 mm Hg (SD 10.62) during chest physiotherapy with Entonox, compared with 10.5 mm Hg (SD 10.4) with oxygen 100% (P greater than 0.02). A further nine mechanically ventilated patients with severe head injuries were given Entonox without chest physiotherapy. There was a mean increase in i.c.p. of 3.8 mm Hg (SD 2.4) (P less than 0.001) when Entonox was given, and a mean decrease of 4.6 mm Hg (SD 2.8) when the nitrous oxide was withdrawn. End-tidal carbon dioxide concentration showed almost no change during nitrous oxide administration (decrease of 0--0.1%). We conclude that nitrous oxide causes an increase in i.c.p. in patients with severe head injuries and exacerbates the increases in i.c.p. occurring during chest physiotherapy.


Programmable calculator: a program for use in the intensive care unit. A program has been prepared for use in a small programmable calculator to allow the rapid and accurate determination of derived data. Changes in oxygen availability, deadspace and shunt fraction can be obtained in a few seconds and therapy altered accordingly. FIO2 required to produce a desired PaO2 can be obtained using the iso-shunt concept.


A double-blind comparison of a novel indanone diuretic (MK-196) with hydrochlorothiazide in the treatment of essential hypertension. 1 The antihypertensive effect and tolerability of MK-196 (10 mg and 15 mg daily) was compared to hydrochlorothiazide (HCT; 50 mg daily) in a 4-week multiclinic, double-blind study involving 42 patients with mild to moderate, essential hypertension. 2 Both doses of MK-196 were as effective and sometimes more effective than HCT in lowering standing and supine systolic and diastolic blood pressures. 3 HCT consistently brought about significant increases in serum uric acid and significant decreases in serum potassium; both doses of MK-196 produced similar but less frequent and smaller changes in both of these parameters. 4 Both doses of MK-196 brought about significant decreases in body weight at Weeks 3 and 4 of drug treatment. 5 Patients taking MK-196 reported fewer adverse clinical reactions (10 mg = 15%; 15 mg = 13% than those taking HCT (21%). 6 MK-196 may offer a therapeutic advantage over HCT as an antihypertensive agent for use in the treatment of mild to moderate, essential hypertension.


Tumour-necrosis factor from the rabbit. III. Relationship to interferons. Tumour-necrosis factor (TNF) is growth-inhibitory or cytotoxic to certain tumour cell lines, and is present in the serum of rabbits injected i.v. with BCG and endotoxin 2 weeks apart (TNF serum). TNF serum also has interferon activity, and as TNF and interferons have a number of properties in common their relationship has been investigated further. TNF was assayed by cytotoxicity in vitro against L cells and interferon by a CPE-inhibition assay with Semliki Forest virus.TNF appears not to be an interferon, on the following bases:1. TNF activity could be separated from the Type I interferon of TNF serum by passage through a Cibacron blue-agarose column or by sequential salt precipitation, ion-exchange chromatography and gel filtration.2. Preparations of Type I interferon induced by poly I, poly C or virus lacked TNF activity.3. Though it was not possible to compare TNF with rabbit Type II interferon (as methods used to induce Type II interferon in other species were unsuccessful in the rabbit) rabbit TNF has a number of properties which distinguish it from the Type II interferons of other species.4. Rabbit TNF inhibited the growth of a human melanoma cell line, and also had effects on certain mouse and rabbit cell lines, whereas the anti-cellular effects of interferons are reported to be species-specific.


Response of T lymphocytes to phytohaemagglutinin (PHA) and to cancer-tissue-associated antigens, measured by the intracellular fluorescence polarization technique (SCM test). Human peripheral-blood mononuclear cells, separated by Isopaque-Ficoll flotation and E-rosette formation, were tested by the fluorescein fluorescence polarization method of Cercek & Cercek (the SCM test). The response to stimulation with PHA or cancer tissue leads to a decreased polarization value TP). The responding cells were present in the T-cell fraction (E-rosette-forming cells), which contained less than 10% macrophages and less than 1% cells with surface-bound Ig. Control experiments with the non-T-cell fraction gave different response patterns. The response of T cells from apparently healthy donors and patients with and without cancer were compared. All of the group of 16 healthy persons had a polarization value (P) which decreased (mean +/- s.e. = 23% +/- 2) after PHA stimulation, compared with no or little decrease after stimulation with cancer tissue, giving cancer indices (P cancer/PPHA) of 1.15--1.56. In 13 patients with carcinoma of the colon, stimulation with PHA produced little decrease of polarization, while stimulation with colonic cancer tissue decreased the polarization in all cases (mean +/- s.e. = 25% +/- 2). The corresponding cancer indices were 0.61--0.86. Seven of 10 colonic-cancer patients tested against ovarian cancer tissue did not respond, whilst 3 patients in this group responded and had a cancer index less than 1.0. Three patients with non-malignant diseases had response patterns similar to those of healthy persons, except for the lack of PHA response in the patient with ulcerative colitis. This method seems to open up new possibilities for evaluation of cancer patients, although further studies including many more patients are needed before any conclusion can be drawn as to the validity of the test.


Radiosensitization of E. coli B/r by 9-anilinoacridines. Six anilinoacridine derivatives have been tested for the ability to act as radiosensitizers. Two gave good sensitization at concentrations of 100 microM or less. Both of these are known to possess significant activity against experimental tumours, and one (m-AMSA) is in Phase II clinical trial as a chemotherapeutic drug. Anilinoacridines may have potential as drugs with both a chemotherapeutic and radiosensitizing role. In spite of their structural similarity, the 2 derivatives which sensitize do so by different mechanisms. Compound VI behaves like a typical hypoxic cell sensitizer but Compound I (m-AMSA) interferes with the accumulation of sublethal damage in either the presence or absence of O2. The latter also displays a post-irradiation sensitizing effect. Differences in mechanism may be related to the relative DNA-binding abilities and electronic differences between the 2 drugs.


Reduced leucocyte alkaline phosphatase activity and decreased NBT reduction test in induced iron deficiency anaemia in rabbits. Iron deficiency anaemia was induced in rabbits by repeated bleeding. The leucocyte alkaline phosphatase (LAP) of 26 +/- 28 units was significantly reduced compared with control values of 233 +/- 35 units (P less than 0.001). Leucocyte NBT reduction was also diminished, both in Hanks solution (P less than 0.01) and in autologous serum (P less than 0.001). After administration of iron, these values returned to normal. The results suggest that reduced LAP may reflect a deficiency of iron dependent constituents which are necessary for the integrity of normal granulocyte metabolism.


The composition of amniotic fluid in pregnancies complicated by fetal anencephaly or spina bifida. The concentration of amniotic fluid glucose, total protein, sodium, potassium, calcium, magnesium and phosphate as well as amniotic fluid osmolality were measured in normal second trimester pregnancies and in second trimester pregnancies complecated by fetal anencephaly or spina bifida. Amniotic fluid glucose concentration was low in anencephaly, while phosphate and total protein were high; calcium concentration and osmolality were slightly elevated with spina bifida. The application of these findings to antenatal diagnosis are discussed.


Indomethacin in the treatment of primary dysmenorrhoea. Thirty-two patients were treated with placebo tablets or indomethacin (25 mg three times daily) in a six-month, double-blind, cross-over trial. During indomethacin therapy, 75 per cent of patients experienced significant pain relief while associated vomiting and diarrhoea were relieved in 44 per cent and 64 per cent of patients respectively. The efficacy of indomethacin was comparable to that of other prostaglandin synthetase inhibitors.


Results of the colposuspension operation for incontinence and prolapse. One hundred and eighty female patients complaining of stress incontinence were treated by a modified Burch colposuspension. Follow-up results were available at six months in 146 patients, at one year in 88 patients and at two years in 43 patients. The overall objective cure for stress incontinence was 87 per cent at six months, 87.5 per cent at one year and 86 per cent at two years. No relationship was found between the cure of incontinence and either the time taken to resume spontaneous voiding after surgery or concomitant hysterectomy. Cystourethrocele was effectively corrected by this procedure.


Double blind trial of ritodrine and placebo in twin pregnancy. In a double blind trial, 25 patients with twin pregnancy were given 40 mg of ritodrine hydrochloride by mouth daily and 24 similar patients received a placebo. The ritodrine group had no significant prolongation of pregnancy nor increase in birth weight, and a high incidence of side effects occurred.


Clinical trial of a new oral contraceptive pill containing the natural oestrogen 17 beta-oestradiol. The natural oestrogen, 17 beta-oestradiol, has been shown not to depress fibrinolysis and apparently has less influence on liver function and lipid metabolism than ethinyl oestradiol, the synthetic oestrogen in conventional 'combined' oral contraceptive tablets. A triple-blind study was therefore made of 215 women during 2051 treatment cycles with oral contraceptives containing either (i) 4 mg of micronized 17 beta-oestradiol and 3 mg norethisterone (Netagen 403), (ii) 4 mg 17 beta-oestradiol plus 2 mg of oestriol and 3 mg norethisterone (Netagen 423) or (iii) 50 microgram ethinyl oestradiol and 3 mg norethisterone (Netasyn). There were no pregnancies or thrombotic incidents. The numbers discontinuing treatment were about the same in the three groups, the main reasons being intermenstrual spotting in those on Netagen 423, amenorrhoea and weight gain in those on Netagen 403 and nausea and weight gain in those on Netasyn. The natural oestrogen showed promise as a new and safe component of the 'combined' pill.


A double-blind trial of single-dose chemoprophylaxis with co-trimoxazole during vaginal hysterectomy and repair. Twenty-five patients received 10 ml co-trimoxazole (800 mg sulphamethoxazole and 160 mg trimethoprim) and 25 a placebo by intravenous infusion during one hour from the commencement of surgery for vaginal hysterectomy with anterior colporrhaphy and posterior colpo-perineorrhaphy. Single-dose chemoprophylaxis with co-trimoxazole was shown to be effective in reducing the incidence of postoperative febrile morbidity and urinary tract infection, especially that caused by Proteus species.


Marcaine, a selective inhibitor of eucaryotic aminoacylation. The effects of marcaine, a myotoxic drug, on the aminoacylation of transfer ribonucleic acid (rRNA) have been studied. The drug is a potent inhibitor of the acylation of rat liver tRNA with leucine and isoleucine but is only mildly inhibitory (or not inhibitory) to acylation with a number of other amino acids which were tested. Further, marcaine inhibited aminoacylation in cell-free systems using components from several mammalian tissues, including muscle, from yeast, and from wheat germ. No effect of the drug was observed in aminoacylation systems from several bacterial species which were tested. The drug inhibits acylation with leucine and isoleucine competitively but exhibited noncompetitive kinetics when the concentrations of adenosine 5'-triphosphate (ATP) and tRNA were varied. Marcaine was also a competitor of leucine in the ATP--pyrophosphate exchange reaction. Two structural analogues of marcaine, carbocaine and xylocaine, also inhibited acylation of rat liver tRNA with leucine but in a noncompetitive fashion. On a molar basis, marcaine appears to be the most effective inhibitor of the three drugs tested.


In vitro transcription by wheat germ ribonucleic acid polymerase II: effects of heparin and role of template integrity. Double-stranded deoxyribonucleic acid (DNA) from bacteriophage lambda is a good template for wheat germ DNA-dependent ribonucleic acid (RNA) polymerase II. We delineated conditions for obtaining maximum polymerase activity using as template both the relatively intact DNA extracted from the the lambda phage and DNA into which single-strand nicks have been introduced by deoxyribonuclease (DNase) I digestion. The deliberate introduction of nicks produces a modest increase in transcription. The NaCl and MgCl2 optima are broader with the nicked template, so that higher concentrations of these salts are needed before polymerase activity begins to decline. Heparin inhibits initiation but not elongation by wheat germ polymerase. Polymerase can be protected against heparin inhibition by forming binary complexes with the template. The formation of these complexes is reduced at low temperature. The complexes, once formed, decay in the presence of heparin with a half-life of 10--20 min. The number of complexes is highly dependent on the degree of nicking of the template, suggesting that single-strand nicks are the predominant type of site where these heparin-resistant complexes are formed. Our data do not allow us to decide whether or not the presence of nicks plays as decisive a role in the absence of heparin.


On the structure and conformational dynamics of yeast phenylalanine-accepting transfer ribonucleic acid in solution. The solution structure of yeast tRNAPhe was investigated by using ethidium as a fluorescent probe in the D loop and the anticodon loop. For this purpose the dihydrouracils in position 16/17 and wybutine in position 37 were substituted by ethidium. The lifetimes and the time-dependent anisotropy of ethidium fluorescence were measured by pulsed nanosecond fluorometry. The kinetics of the transitions between different states of the tRNAPheEtd derivatives were determined by chemical relaxation measurements. It was found that the ethidium label irrespective of its position exhibits three different states called T1, T2 and T3 characterized by lifetimes tau 1 = 30 ns, tau 2 = 12 ns, and tau 3 = 3 ns. The lifetime differences are due to different accessibilities of ethidium for solvent quenching in the three states. Thus, there are three different defined structural environments of the ethidium in both the anticodon and the D loop. The distribution of the three states was measured as a function of Mg2+ concentration and temperature; it was found that state T3 is favored over states T2 and T1 by both increasing Mg2+ concentration and increasing temperature. The chemical relaxation kinetics exhibit a fast transition between T1 and T2 (10--100 ms) and a slow transition between T2 and T3 (100--1000 ms). The rates of both transitions depend likewise on Mg2+ concentration and temperature. The equilibrium and kinetic data clearly show the presence of strong and weak interactions between Mg2+ and tRNA. A cooperative model accounting for this behavior is developed. The ethidium probe behaves identically when located in different regions of the tRNA regarding both its distribution of states and its transition kinetics. This suggests that the different spectroscopic states report different conformations of the tRNA structure. The dependence of the three states on Mg2+ and spermine indicates that conformation T3 is closely related to or identical with the crystal structure. The rotational diffusion constants indicate that of all three states T3 is most extended while T2 is most compact. The thermodynamic analysis reveals that the strongly bound Mg2+ ions reduce both the activation entropy and enthalpy of all transitions. The weakly bound Mg2+ ions increase both the activation enthalpy and entropy of the slow transition between T2 and T3. It is suggested that the breaking of several intramolecular bonds, e.g., hydrogen bonds, is involved in this transition.


Amino acid sequence of the ribosomal protein L21 of Escherichia coli. The primary structure of protein L21 from the 50S subunit of Escherichia coli ribosomes has been completely determined by sequencing the peptides obtained by digestion of L21 with trypsin before and after modification of the arginine residues with 1,2-cyclohexanedione, Staphylococcus aureus protease, thermolysin, and pepsin. Automated Edman degradation using a liquid-phase sequenator was carried out on the intact protein as well as on a fragment arising from cleavage with cyanogen bromide. Protein L21 consists of a single polypeptide chain of 103 amino acids of molecular weight 11 565. An estimation of the secondary structure of protein L21 and a comparison with other E. coli ribosomal protein sequences are presented.


Suicide inactivation of bacterial cystathionine gamma-synthase and methionine gamma-lyase during processing of L-propargylglycine. L-Propargylglycine, a naturally occurring gamma, delta-acetylenic alpha-amino acid, induces mechanism-based inactivation of two pyridoxal phosphate dependent enzymes of methionine metabolism: (1) cystathionine gamma-synthease, which catalyzes a gamma-replacement reaction in methionine biosynthesis, and (2) methionine gamma-lyase, which catalyzes a gamma-elimination reaction in methionine breakdown. Biphasic pseudo-first-order inactivation kinetics were observed for both enzymes. Complete inactivation is achieved with a minimum molar ratio ([propargylglycine]/[enzyme monomer]) of 4:1 for cystathionine gamma-synthase and of 8:1 for methionine gamma-lyase, consistent with a small number of turnovers per inactivation event. Partitioning ratios were determined directly from observed primary kinetic isotope effects. [alpha-2H]Propargylglycine displays kH/kD values of about 3 on inactivation half-times. [alpha-3H]-Propargylglycine gives release of tritium to solvent nominally stoichiometric with inactivation but, on correction for the calculated tritium isotope discrimination, partition ratios of four and six turnovers per monomer inactivated are indicated for cystathionine gamma-synthase and methionine gamma-lyase, respectively. The inactivation stoichiometry, using [alpha-14C]-propargylglycine, is four labels per tetramer of cystathionine gamma-synthase but usually only two labels per tetramer of methionine gamma-lyase (half-of-the-sites reactivity). Two-dimensional urea isoelectrofocusing/NaDodSO4 electrophoresis suggests (1) that both native enzymes are alpha 2 beta 2 tetramers where the subunits are distinguishable by charge but not by size and (2) that, while each subunit of a cystathionine gamma-synthase tetramer becomes modified by propargylglycine, only one alpha and one beta subunit may be labeled in an inactive alpha 2 beta 2 tetramer of methionine gamma-lyase. Steady-state spectroscopic analyses during inactivation indicated that modified cystathionine gamma-synthase may reprotonate C2 of the enzyme--inactivator adduct, so that the cofactor is still in the pyridoxaldimine oxidation state. Fully inactivated methionine gamma-lyase has lambda max values at 460 and 495 nm, which may represent conjugated pyridoximine paraquinoid that does not reprotonate at C2 of the bound adduct. Either species could arise from Michael-type addition of an enzymic nucleophile to an electrophilic 3,4-allenic paraquinoid intermediate, generated initially by propargylic rearrangement upon a 4,5-acetylenic pyridoximine structure, as originally proposed for propargylglycine inactivation of gamma-cystathionase [Abeles, R., & Walsh, C. (1973) J. Am. Chem. Soc. 95, 6124]. It is reasonable that cystathionine gamma-synthase is the major in vivo target for this natural acetylenic toxin, the growth-inhibitory effects of which are reversed by methionine.


Conformation transitions of a tRNA--aminoacyl-tRNA synthetase complex induced by tRNAs bearing different modifications in the 3' terminus. The influence of modifications of the 3'-terminal adenosine of tRNAPhe (yeast) on the complex formation between this tRNA and phenylalanyl-tRNA synthetase (yeast) has been investigated by using fluorescence titrations and fast kinetic techniques. Subtle changes in the 3' terminus are reflected by distinct alterations in the two-step recognition process which had been demonstrated earlier for the native substrate tRNAPheCCA [Krauss, G., Riesner, D., & Maass, G. (1977) Nucleic Acids Res. 4, 2253--2262]. Binding experiments with tRNAPheCC, tRNAPheCCA-ox-red, tRNAPheCC2'dA, tRNAPheCC3'dA, tRNAPheCC-formycin, and tRNAPheCC-formycin-ox-red confirm that the 3'-terminal adenosine participates in a conformational change of the tRNA--synthetase complex. This is valid in both the absence and presence of phenylalaninyl-5'-AMP, the alkyl analogue of the aminoacyladenylate. As compared to tRNAPheCCA, a slower conformational change is observed with the competitive inhibitor tRNAPheCC-formycin-ox-red. The reaction enthalpy and/or the quench of the Y-base fluorescence that accompany the conformational change are altered upon binding of tRNAPheC2'dA, tRNAPheCC3'dA, and tRNAPheCC-formycin. It is evident that the final adaptation between tRNA and its synthetase in the complex is determined by the chemical nature of the 3'-terminal nucleotide. This is of vital importance for the specificity of the aminoacylation process.


Affinity labeling of the 3'-OH terminal binding site of the ribonucleic acid chain on deoxyribonucleic acid dependent ribonucleic acid polymerase from Escherichia coli. Nucleoside triphosphates modified at the 3'-OH are chain terminators for RNA polymerase. They form inactive ternary complexes with the enzyme, poly(dT), and oligoadenylate, the stabilities of which depend upon the length of the oligonucleotide. Employing [5'-32P]p(Ap)10A, together with the reactive analogues 3'-(bromoacetamide)-3'-deoxyadenosine triphosphate or 3'-(isothiocyanato)-2',3'-dideoxyadenosine triphosphate, as well as 3'-amino-3'-deoxyadenosine triphosphate, followed by cross-linking with glyoxal, we labeled RNA polymerase primarily at the beta' subunit. The latter therefore appears to contain at least in part the 3'-OH terminus of the nascent RNA chain when the enzyme is in the form of the ternary complex.


Modification of deoxyribonucleic acid by a diol epoxide of benzo[a]pyrene. Relation to deoxyribonucleic acid structure and conformation and effects on transfectional activity. The effects of secondary structure on DNA modification by (+/-)-7 beta, 9 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9,10-tetrahydrobenzol[a]pyrene [(+/-)BPDE I] were investigated. No differences in the total extent of (+/-) BPDE I binding to double- and single-stranded calf thymus DNA were found. High-performance liquid chromatography (LC) of the nucleoside adducts obtained from hydrolysates of native and denatured calf thymus, as well as from superhelical and linear plasmid DNA, indicated that in all cases the major adduct (60--80% of total adducts) was formed by reaction of the (+) enantiomer of BPDE I with the N-2 position of dG residues in the DNA. A minor adduct formed from the reaction of the (-) enantiomer with dG residues was also detected and was present in greater amounts in denautred DNA than in native DNA. Small amounts of BPDE I--dA and BPDE I--dC adducts were also detected in both the single- and double-stranded DNAs. Restriction enzyme analysis of BPDE I modified SV40 and phage lambda DNA provided evidence that the modification of DNA by this carcinogen is fairly random with respect to nucleotide sequence. Partial hydrolysis of modified plasmid DNA by the single-strand-specific S1 nuclease and LC analysis of the nucleoside adducts in the digested and undigested fractions of the DNA revealed no preferential excision by the S1 nuclease of the different BPDE I--deoxynucleoside adducts. Functional changes in BPDE I modified DNA were demonstrated. With increasing extents of modification, there was a decrease in the ability of plasmid DNA to transfect a receptive Escherichia coli strain to antibiotic resistance.


The effect of magnesium starvation on the dissociation of ribosomal proteins from Escherichia coli K-12 ribosomes. The effect of magnesium starvation upon the fate of individual ribosomal proteins was studied in Escherichia coli. During a 21 h incubation in the absence of Mg2+ the 30 S subunit was more susceptible to degradation, retaining an average 31.9% of its ribosomal proteins as compared to 40.0% for the 50 S subunit. An examination of those 50-S proteins dissociated to a lesser extent than the average value (L1, L2, L3, L7, L10, L13, L16, L17, L19, L21, L22, L23, and L29) revealed that, with the exception of L16, all were classified by Dohme and Nierhaus [5] as tightly bound. Of the ribosomal proteins dissocated during magnesium starvation only five were reincorporated (and these to a minimal degree) during recovery of cells in a medium containing Mg2+. These studies suggest that ribosomal proteins once released from the ribosome particles during magnesium starvation are not reutilized in the assembly of new subunits.


Escherichia coli mut T1. II. Consequences of modification on the association of DNA with the cell membrane. 1. Two isogenic strains of Escherichia coli, K-12 which differ by mutator gene character (mut T1) have been studied. This characteristic caused introduction of a high frequency of undirectional transversions, A-T leads to -CG, into the DNA of the strain harboring it. 2. It had been previously shown that the presence of this gene is accompanied by an alteration of a cell membrane component. Now, the nuclease susceptibility of DNA associated with membrane/DNA/DNA polymerase complexes is reported. DNA of mut T1 membranes is more sensitive towards exogenous nuclease than DNA of membrane complexes from the wild type mut+ strain. 3. Auto-digestion of this DNA by endogenous nuclease associated with the membrane complex is, also, more severe in preparations derived from mut T1 than from the wild-type strain, mut+, but to a lesser extent than observed with exogenous nucleases. 4. Nuclease susceptibility of mut+ membrane bound DNA is markedly influenced by the growth state of the cell. The nuclease susceptibility of membrane bound DNA from mut T1 cells, however, shows no differences between stationary and log states. 5. These differential sensitivities may be due to conformational changes in the membrane introduced as a pleiotrophic consequence of an altered membrane protein. A pertinent role of this protein in a modified replication/repair complex is an attractive suggestion, especially in the context of the mutator character of this strain.


Host-cell reactivation of alkylated T7 bacteriophage. Purified T7 phage, treated with methyl methanesulfonate, was assayed on Escherichia coli K-12 host cells deficient in base excision repair. Phage survival, measured immediately after alkylation or following incubation to induce depurination, was lowest on a mutant defective in the polymerase activity of DNA polymerase I (p3478). Strains defective in endonuclease for apurinic sites (AB3027, BW2001) gave a significantly higher level of phage survival, as did the strain defective in the 5'--3' exonuclease activity of DNA polymerase I (RS5065). Highest survival of alkylated T7 phage was observed on the two wild-type strains (AB1157, W3110). These results show that alkylated T7 phage is subject to repair via the base excision repair pathway.


Two forms of pyruvate kinase in Escherichia coli. A comparison of chemical and molecular properties. The two forms of pyruvate kinase (ATP:pyruvate 2-O-phosphotransferase, EC 2.7.1.40) present in Escherichia coli have been purified from the same cultures and crystallized. A modified procedure for the purification of type I pyruvate kinase is described. Molecular weight, subunit structure, amino acid composition, NH2-terminal amino acid, maps of tryptic peptides and conditions for crystallization have been determined for the two forms. A comparison of these data shows that the two forms are different proteins, each being a tetramer of identical subunits.


The thermostability of glucoamylase immobilized in different ways has a certain limit. Thermostability of glucoamylase from Aspergillus niger is increased both by immobilization and substrate binding. However, the total superimposed stabilization effect at a given condition is apparently restricted by a certain limit which hardly depends on the mode of immobilization of the enzyme, and is determined mostly by the enzyme-substrate complex formation.


Spatial proximity of a tyrosyl and a lysyl residue in the active site region of yeast 3-phosphoglycerate kinase. The effect of 7-chloro-4-nitrobenzofurazan on yeast 3-phosphoglycerate kinase causes a modification of one tyrosyl residue concomitantly with a total loss of activity of the enzyme. The modification is not accompanied by any significant conformational change. A total protection against inactivation is observed with the substrates : furthermore, AMP, tripolyphosphate and pyrophosphate afford an effective protection. At pH 9, a shift in the absorbance spectrum of the tyrosine O-nitrobenzofurazan derivative of 3-phosphoglycerate kinase is observed. It can be related to the transfer of the reagent from tyrosine to lysine. The N-nitrobenzofurazan derivative is also completely inactive. It is concluded that a lysine residue is located close to the essential tyrosyl residue.


Methylated amino acids in ribosomal proteins from Escherichia coli treated with ethionine and from a mutant lacking methylation of protein L11. In the present study, the nature, proportions and distribution of methylated amino acids in ribosomal proteins from Escherichia coli grown in the presence of ethionine and from mutant prm 1 were studied. The undermethylated ribosomes had been labeled by addition in vitro or in vivo of radioactive methyl groups from S-adenosylmethionine or from methionine. The following compounds were identified : N alpha-mono-, di- and trimethylalanines, N epsilon-mono-, di- and trimethyllysines, methylamine and N alpha-trimethylalanyllysine. Except for the latter compound and N-alpha-dimethylalanine, all other derivatives had been previously identified in the literature. It is shown that the dipeptide had been in the past mistaken for N epsilon-monomethyllysine, and arises through incomplete hydrolysis in 24 hrs of the N-terminal peptide bond of protein L11. The results of the present study are discussed in the light of previous work on ribosomal protein methylation by the authors and other workers in the field.


[Effect of luliberin on the activities of mitochondrial respiratory enzymes]. Luliberin, a luteinizing hormone-releasing hormone, was shown to inhibit the respiratory enzymes of rat liver mitochondria and submitochondrial particles prepared from beef heart mitochondria. At the hormone concentration of 8.10(-6) M the NADH-oxidase activity of the submitochondrial particles was inhibited by 50%. The fragments of the hormone and its analogs and pyroglutamic acid, oxytocin and bradikinin possessed practically no inhibiting effects. In the case of submitochondrial particles the inhibition was only observed in the presence of Ca2+ and was significantly decreased after addition of bovine serum albumin and phospholipase inhibitors -- butacaine and dicaine. It is assumed that the effect of luliberin on the respiratory chain is mediated through mitochondrial phospholipase.


[Kinetics of phenylacetamide hydrolysis by immobilized penicillinamidase]. The kinetics of phenylacetamide hydrolysis catalyzed by polyacrylamide gel-immobilized penicillinamidase were studied. The Km and Kp values obtained were compared to the literary data for the specific substrate--benzylpenicillin. It was shown that the type of inhibition by the reaction product was the same, whereas the efficiency of binding of phenylacetic acid depended on the substrate structure.


[Phospholipids of E. coli and activity of alkaline phosphatase]. The effects of liposomes prepared from the E. coli lipids on the activity of soluble alkaline phosphatase and on the complementation reaction between its subunits were studied. It was shown that the liposomes nonspecifically catalyze the dimerization of the enzyme subunits without changing the dimer activity. The effects of phospholipases A2 and C on the activity of membrane-bound alkaline phosphatase were studied. An interrelationship was found between the level of hydrolysis of membrane phosphatidyl glycerol (PG) by these enzymes and the changes in the activity of membrane-bound alkaline phosphatase. It was also shown that PG is less accessible to the effects of phospholipases in the cells with derepressed biosynthesis of alkaline phosphatase. It is assumed that the membrane PG interacts with the membrane-bound alkaline phosphatase during its translocation into the periplasm.


[Selective inhibition of ribosomal RNA synthesis by rifampicin in E. coli cells]. Under partial inhibition of total RNA synthesis by rifampicin the formation of beta- and beta'-subunits of RNA polymerase is stimulated and the rRNA synthesis is selectively repressed. The differential rate of synthesis of the beta- and beta'-subunits increases from 1,15% up to 2,88% in the presence of 30 micrograms rifampicin per ml. Simultaneously the differential rate of rRNA synthesis decreases from 41% down to 10%. The degree of inhibition of rRNA synthesis by rifampicin depends on the cell growth rate.


Antibacterial activity of seawater and microvibrion-predators (Microvibrio marinus Roscoffensis) in it. In the seawater of the Atlantic Ocean and surface waters of different points of the World Ocean, including arctic basins and equatorial regions, microvibrion-predators of bacterial cells have been discovered. The number of microvibrions in the surface waters of the English Channel reaches a thousand cells per ml, but it varies, depending on the season. In this article a method of the separation and culture of microvibrions is described. The ultrastructure of two strains of microvibrions was investigated and a similarity found between their structure and Gram-negative bacteria. The distinctive features of microvibrions are their sea derivation and the ability to reproduce only in the presence of bacteria, without penetrating bacterial cells. To a certain extent, the antibacterial activity of seawater is connected with the presence in it of microvibrions.


Physiological and biochemical properties and morphology of Saccharomyces cerevisiae VKMu-488 cells incorporated into polyacrylamide gel. The enzymatic activity, viability, respiratory activity, and ultrastructural changes in saccharomyces cerevisiae VKMu-488 cells, which carry out the stereospecific 17 beta-reduction of methyl esters, was studied. The 17 beta-hydroxysteroid dehydrogenase activity of yeasts in gel is four times lower than that of free cells and is unstable. The decrease in the viability and respiratory activity immediately after immobilization, the disturbance in the ultrastructure of the cells in gel along with the progressive lysis of the cells in the course of the transformation indicate that polymerization has a stressful effect on this culture. It was found that the immobilized yeasts can grow on the surface of the gel in the presence and absence of nutrient medium. A single incubation of granules containing cells in nutrient medium greatly stabilizes the original activity of the immobilized cells. The activation and stabilization of the activity are probably due to the participation of a heterogeneous population in the transformation: the original population incorporated into the gel and the new population which grows in the gel after immobilization as well as to the stability of the ultrastructural organization of this mixed population in the course of repeated transformations of secoketone.


Balance of DNA and protein syntheses, processes of stabilization and repair of breaks in DNA strands, and cell viability. The dependence of the stabilization and repair of breaks in the DNA strands on the balance of the syntheses of DNA and protein and the relationship of these processes to the viability of E. coli cells were experimentally substantiated. It was established that equilibrium of the rates of DNA and protein syntheses is a necessary condition for the functioning of processes of DNA repair. "Imbalanced" breaks in the DNA strands in vivo are a consequence of a sharp decrease in the effectiveness of DNA repair processes.


Properties of penicillin amidase immobilized by copolymerization with acrylamide. An enzyme preparation in a spherical granule form was obtained by copolymerization of penicillin amidase (EC 3.5.1.11) (previously modified with maleic anhydride) and acrylamide via a crosslinking agent. As compared with the native enzyme, immobilized amidase is more resistant to heating, has a lower affinity to benzylpenicillin, and is less inhibited by phenylacetate. Its substrate specificity and optimum pH remain unchanged.


[Effect of presynaptic neurotoxin notechis II-5 from tiger snake venom on the motor nerve endings of mice]. The neurotoxin notechis II-5 (N-II-5) from tiger snake venom (Notechis scutatus) induces three-phasic changes in miniature end-plate potential (MEPP) frequency recorded in the mouse diaphragm muscle: an initial fall of frequency followed by increase and decrease in MEPP frequency up to complete blockade. The effect of N-II-5 was enhanced with rising of the solution temperature from 20 to 30 and 35 degrees C. Removal of Ca2+ from the solution prevented the presynaptic effect of N-II-5. After washing out of the muscle from N-II-5 with Ca-free solution, addition of Ca2+ to the solution provoked the development of the effect typical of the effect of typical of the toxin. In the presence of N-II-5 an increase in K+ concentration in the solution up to 20 mM did not result in a sharp rise of MEPP frequency characteristic of depolarized nerve terminals. The agents that raise Ca2+ axoplasmic concentration not on account of depolarization of nerve terminals (hypertonic solution, ionofor A23187) preserved the capacity for increasing MEPP frequecy. It is suggested that the presynaptic effect of N-II-5 is related to its phospholipase activity and can be explained by disturbance of the activity of release sites rather than by depletion of transmitter stores.


[Change of toxic and antineoplastic properties of ftorafur by means of action on nonspecific microsomal oxidase]. The inducers of microsomal hydroxylases, phenobarbitone and methylcholanthrene, inhibited the development of neurotoxic shock provoked by high doses of ftorafur in mice, but stimulated the animal mortality on the 4th-8th day after the drug administration. The opposite effect on both toxicity manifestations has been obtained under the action of the inhibitor SKF 525-A. Pretreatment of the animals with phenobarbitone or phenobarbitone-methylcholanthrene combination markedly increased the antineoplastic activity of ftorafur determined by a loss of the spleen weight in mice infected with Rauscher's leukemia.


[Restoration of antineoplastic reactivity of the leukocytes after preincubation]. Specific antitumor sensitization in patients with carcinoma of the stomach was revealed by means of the leukocyte migration inhibition test. After 24-hour preincubation at 4 degrees C in a serum-free medium the areactive leukocytes from patients with carcinoma of the stomach (stages III--IV) acquired the ability to react specifically to allogeneic antigens of the tumor of the same localization. Preincubation did not influence the inhibition of migration of leukocytes in non-tumor patients by stomach carcinoma antigens. The supernatant of preincubated leukocytes of tumor patients contained substances that inhibited migration of indicator leukocytes.


[Immunofluorescence studies of the immunoglobulin receptors of B-lymphocytes]. The surface membrane Ig receptors of B lymphocytes in patients with atopic pollen asthma were studied by the direct immunofluorescence method. Use was made of labelled sera against human IgE, and IgG. It was shown that on the average 2.1 +/- 0.33% lymphocytes had IgE receptors, 8.4 +/- 0.63% IgG receptors and 18.7 +/- 1.16% human globulin receptors. A specific nature of B lymphocytes is suggested.


[Indirect immunoelectron microscopic method of localization of albumin in mouse hepatocytes]. The mouse liver was fixed by 12 fixator variants on the basis of glutaraldehyde, paraformaldehyde, picric acid and lysin, and the localization of albumin was studied by the indirect immunoperoxidase method. The comparison of the results shows that immersion in 8% paraformaldehyde is a preferable method of fixation. Albumin was detected on ribosomes and membranes of the nuclear envelope, the coarse endoplasmic reticulum and in the matrix of the hepatocytic cytoplasm. It is suggested that albumin synthetized is directly discharged into cytosol and then into the extracellular space.


Processing by the thymus is not required for cells that cure and populate W/WV recipients. Adult marrow, fetal liver or nu/nu mouse marrow from histocompatible donors was grafted into genetically anemic W/WV recipients, and all three types of grafts cured thymectomized as well as intact W-anemic recipients. With the latter two types of graft, the genetic anemia was cured by cells that could not have been processed in a mature thymus, since the adult recipients were thymectomized before receiving the grafts, the nu/nu donors were congenitally thymusless, and the fetal donors were used at 16 days of gestation. Chromosome-marked marrow grafts were used to show that immune systems were populated to similar degrees in thymectomized and intact W/WV recipients. Therefore, the cells derived from the donor marrow graft that partially populate the immune systems of W-anemic recipients do not require thymus processing. Small numbers of liver rudiment or yolk sac cells from fetal donors less than 12 days old failed to cure W/WV recipients, even when mixed with adult thymus cells. Therefore, the lack of adequately developed thymic helper cells appears not to be the reason why early fetal hemopoietic stem cells fail to cure W/WV recipients.


Proliferative state of human pluripotent hemopoietic progenitors (CFU-GEMM) in normal individuals and under regenerative conditions after bone marrow transplantation. The proliferative state of human pluripotent hemopoietic progenitors (CFU-GEMM) was assessed in normal bone marrow transplant donors and in recipients after engraftment using the tritiated thymidine suicide method. In contrast to BFU-E and CFU-C, CFU-GEMM were found to be quiescent. In samples obtained during early regeneration after transplantation, CFU-GEMM were significantly reduced after short-term exposure to 3H TdR when compared to controls. This observation suggests active participation of CFU-GEMM in bone marrow regeneration.


Effects of prostaglandins E2 and I2 on human lymphocyte transformation in the presence and absence of inhibitors of prostaglandin biosynthesis. 1 The reactivity of human peripheral blood mononuclear cells to phytohaemagglutinin ([3H]-thymidine incorporation) was enhanced in indomethacin- and eicosatetraynoic acid-treated cells compared with untreated cells, from normal volunteers. This suggests that endogenously synthesized prostaglandins have an inhibitory effect during cell preparation and/or culture. 2 Prostaglandin E2 inhibited [3H]-thymidine incorporation induced by suboptimal phytohaemagglutinin concentrations and had a more potent effect on indomethacin-treated cells than on untreated cells. 3 Prostaglandin I2 also exhibited an inhibitory effect and, under defined conditions, was more potent than prostaglandin E2 or than prostacyclin which had been allowed to decay at pH 7.4 and 37 degrees C. 4 These results indicate that, in attempting to define altered lymphocyte reactivity in disease states, the involvement of prostaglandins should be considered both during cell preparation and culture.


Inhibition of insulin secretion from the perfused pancreas of the rat by pizotifen. 1 In the perfused rat pancreas the effects of pizotifen on insulin release induced by 20 mM glucose were studied. 2 Pizotifen (10 and 100 muM) significantly reduced the insulin release during a 25 min perfusion period to 49% and 7% of the controls. 3 The same concentrations of the structurally related agents cyproheptadine, doxepin, and chlorpromazine produced a comparable inhibition.


Inhibition by pentobarbitone and urethane of the in vitro response of the adenohypophysis to luteinising hormone-releasing hormone in male rats. 1 The effect of urethane (ethyl carbamate) and sodium pentobarbitone on the luteinising hormone-releasing hormone (LH-RH)-stimulated secretion of luteinising hormone (LH) was investigated with hemipituitaries obtained from male rats and incubated in vitro. 2 Urethane and pentobarbitone were added to the incubation medium to provide final concentrations of 2.2 mg or 4 micrograms/ml and 100 micrograms or 0.1 microgram/ml respectively. The high doses of these anaesthetics blocked the LH-RH stimulated secretion of LH. The low doses significantly reduced the amounts of LH released in response to LH-RH but did not block the response completely. 3 Both concentrations of urethane reduced the basal release of LH. 4 The inhibitory action of the anaesthetics was reversible. 5 The results indicate that the two anaesthetics most commonly used in neuroendocrine experiments have a significant inhibitory action on the release of LH from the adenohypophysis.


High dose flupenthixol decanoate in chronic schizophrenia. In a double-blind trial, female 'drug-resistant' chronic schizophrenic in-patients were given high dose or standard dose flupenthixol decanoate for 13 weeks. Plasma flupenthixol levels showed a five-fold interindividual variation, but were consistently higher with the high dose. Analysis of final scores showed no statistically significant differences between groups with regards to mental state, ward behaviour and extrapyramidal side-effects. When compared with pre-trial scores, the extrapyramidal side-effects worsened significantly in the high dose patients and social withdrawal decreased in the standard dose patients. The mental state of a sub-group of patients, possibly drug resistant for pharmacokinetic reasons, improved significantly on the high dose over the 13 weeks.


Renal function in patients on lithium treatment. A cross-sectional study was performed on 66 lithium-treated patients to investigate a possible changed kidney function, using [51Cr]-EDTA clearance, and urinary excretions of albumin and beta 2-microglobulin. Thirteen patients showed abnormal test results: seven had decreased glomerular filtration rate, four had increased albumin excretion and four had increased excretion of beta 2-microglobulin. There was no correlation between length of treatment with lithium or hypothyroidism (10 patients) and impaired renal function. Four patients had already manifested signs of renal dysfunction before lithium treatment. The high prevalence of impaired renal function among our patients is unexplained but lithium could be one possible cause.


A technique for the difficult peripheral arterial aneurysm. Complete external control of the vessels feeding some aneurysms may be very difficult and occasionally only achieved at the cost of damage to surrounding structures. A method is described in which Foley catheters are introduced into the feeding vessels from within the aneurysm and then cut distal to their clamped side branch to enable Dacron grafts to be passed over them and sutured in place. This technique has been used in 3 patients with very satisfactory results and is offered as an alternative when standard control is hazardous, or impossible.


Preservation of the injured spleen.

Preservation of the injured spleen. Experience with suture repair of splenic lacerations in 14 children and 8 adults is reported. The youngest patient was 3 days old and the oldest 60 years. Adequate healing was confirmed by postoperative splenic scans with or without arteriography. The 2 deaths in this series were not related to the splenic repair and were due to associated major injuries. The course of illness in the remaining 20 survivors did not reveal a failure of this technique or a complication related to the use of this conservative approach. It is expected that salvage of the traumatized spleen will eliminate the risk of fulminant postoperative infections that may be fatal.


Effect of beta-blockers on arrhythmias during six weeks after suspected myocardial infarction. Twenty-four-hour electrocardiographic tape-recording was used to investigate the incidence of arrhythmias in patients with suspected myocardial infarction who were receiving either propranolol, atenolol, or placebo. Recordings begun within 24 hours after admission to a coronary care unit showed that 76% of patients eventually found to have had a myocardial infarction had ventricular arrhythmias of a type generally regarded as serious, whereas of patients in whom myocardial infarction was not substantiated, only 24% had such arrhythmias. At one and six weeks after admission the incidence of arrhythmias ranged from 25% to 33% irrespective of diagnosis. Of patients monitored at both one and six weeks, however, only 5% had arrhythmias on each occasion. Patients treated with propranolol and atenolol showed a similar incidence of arrhythmias to those taking placebo. There was no difference in the incidence or type of arrhythmias recorded between patients who died and those who were still alive at six weeks.These results confirm that "serious" ventricular arrythmias occur in most patients during the acute phase of myocardial infarction and suggest that they do not constitute an independent risk factor. Beta-blockers showed little evidence of useful antiarrhythmic action in the dosage used, but increasing the dosage in suspected myocardial infarction is not practicable because of the risk of hypotension. The findings raise grave doubts about the value of studying arrhythmias to assess drugs intended to reduce mortality from myocardial infarction.


Hats for the newborn infant.

Hats for the newborn infant. The efficacy of a Gamgee-lined hat in reducing the rate of fall in rectal temperature of infants during the first 30 minutes of life was studied. The trial, which included 211 infants, was randomised, prospective, and controlled. One hundred and seven infants were exposed to overhead radiantheaters. Of these, only 30 had normal deliveries, so the analysis was confined to the infants who were not subjected to radiant heat, and in this group no conclusions were drawn about the efficacy of a hat or a radiant heater. In the 104 infants not subjected to radiant heat, body weight, initial rectal temperature, the application of hat, the environmental temperature, and the duration of exposure while naked were all found to influence measureably the rate of fall in rectal temperature during the first 30 minutes. Gamgee-lined hats should be routinely used to minimise heat loss, especially in small infants exposed at birth, during surgical operations, and during investigations necessitating prolonged exposure.


Development of vasoactive intestinal polypeptide (VIP) containing neurones in the rat brain. The development of VIP-containing neurones in the rat CNS and duodenum has been studied using a specific radioimmunoassay and immunohistochemistry. In the brain, VIP immunoreactivity appears entirely postnatally, while VIP in peripheral neurones in the duodenum was present before birth. The developmental changes observed in cerebral cortex appear to represent the maturation of a population of intrinsic cortical interneurones which contain VIP. These neurones develop entirely after birth. They are first seen in deep cortical layers, but later spread out into all cortical layers, particularly layers II--IV. Changes in the intensity of VIP cell body fluorescence can be correlated with changes in VIP content in the cortex measured by radioimmunoassay. Thus VIP forms a unique chemical marker for studying the maturation of a cortical neurone.


Identification of some anaerobic bacteria in nonspecific anaerobic infections in animals. Over 200 anaerobic bacterial isolates were recovered in a veterinary diagnostic laboratory from nonspecific infectious disease from 72 specimens originating from ten animal species. The majority of isolates were nonsporeforming bacteria and about half were identified to species. Bacteroides species formed the major group and included B. oralis, B. fragilis, B. corrodens, B. ruminicola subspecies ruminicola, B. ruminicola subspecies brevis and various subspecies of B. melaninogenicus. Gram-positive anaerobic cocci constituted the next major group of isolates and the main species identified in cattle was Peptococcus indolicus. Clostridial species were uncommon. Nine specimens yielded a pure culture of an anaerobe and, in samples containing mixtures of bacterial species, each specimen yielded an average of 3.1 anaerobic and 1.4 aerobic bacterial species. The failure to identify many of the isolates is discussed.


An epizootic of bovine tuberculosis in Barbados, West Indies. A 57 cow dairy herd, believed to be in excellent condition and producing well, was found to contain 30 tuberculous cattle after cervical skin testing employing bovine PPD tuberculin with the single intradermal comparative test. Testing was initiated after two cows from this herd were found at meat inspection to have tuberculosis. The reliability of bovine PPD tuberculin for testing as well as significant epidemiological, laboratory and meat inspection factors are presented.


Experimental Klebsiella and Salmonella infection in neonatal swine. Twenty colostrum-fed piglets from three sows were separated from the sows 24 hours after birth and were randomly divided into five groups of four piglets each. Every piglet in each of four test groups was orally inoculated with about 10(10) colony forming units of Salmonella typhimurium, Salmonella choleraesuis var Kunzendorf or one of two isolates of Klebsiella pneumoniae. One group served as uninoculated controls. Piglets infected with K. pneumoniae developed severe diarrhea beginning about 12 hours after inoculation. They became dehydrated and weak but continued to drink. There were no morphological alterations in intestinal mucosa when piglets were killed and necropsied 48 or 72 hours after inoculation. Klebseilla pneumoniae was isolated from intestine and feces but not from liver or spleen. Piglets inoculated with S. choleraesuis became lethargic and disinterested in food by 24 hours after inoculation. Diarrhea developed by 48 hours after inoculation. Lesions at necropsy 60 or 72 hours postinoculation were subcutaneous edema, mesenteric lymphadenitis, diffuse intestinal superficial mucosal necrosis with villous atrophy, and focal deep ulceration in the ileum. Salmonella choleraesuis was isolated from all segments of intestine and from feces, liver and spleen. Piglets inoculated with S. typhimurium developed a relatively mild diarrheal disease with lesions similar to those with S. choleraesuis infection but less severe. The inoculated organism was recovered from all areas of intestine and from feces, liver and spleen. Serum from infected and control piglets had high (greater than 1:256) agglutinating titres against S. typhimurium but low titres (0 to 1:8) against S. choleraesuis. The agglutinins were assumed to originate from colostral antibodies.


The effects of the anticonvulsant valproic acid on cerebral indole amine metabolism. The effects of valproic acid (500 mg/kg, ip, 1 h prior to testing) on indole amine metabolism were studied in rats by measurement of the contents of tryptophan, 5-hydroxytryptophan (5-HTP), 5-hydroxytryptamine (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) in the cerebral hemisphere. Tryptophan and 5-HIAA levels were increased, whereas 5-HTP and 5-HT remained unchanged. Furthermore, valproic acid failed to alter the levels of 5-HTP and DOPA, 5-HT and DA, and 5-HIAA in animals pretreated, respectively, with 3-hydroxybenzyl hydrazine (a decarboxylase inhibitor), pargyline (a monoamine oxidase inhibitor), or probenecid (a compound which blocks 5-HIAA transport out of the brain and cerebrospinal fluid). These results militate against the possibility that valproic acid alters the rate of tryptophan hydroxylation or the synthesis of 5-HT. However they do support the concept that valproic acid increases brain 5-HIAA by inhibition of the transport mechanism which removes 5-HIAA from the brain.


A source of large axons for neurophysiology: the North Atlantic squid Illex illecebrosus. A new axon preparation from the ommastrephid squid (Illex illecebrosusus (Lesueur)) is described. This squid is common in the Northwestern Atlantic and features a number of long, unbranched, and moderately large-diameter axons having no apparent 'twigging.' Although the diameters of these axons are somewhat smaller than those of 'giant' axons coming from some other species of squid, this axon preparation should be considered as an attractive alternative for neurobiologic research.


Percutaneous transfemoral retrieval of the "runaway" ventriculoatrial shunt. A 22-year-old man with a ventriculoatrial shunt was admitted to hospital for investigation of headache, nausea and vomiting. During a procedure done to convert the ventriculoatrial shunt to the ventriculoperitoneal type, the shunt tubing was inadvertently cut in the neck. The atrial end was found to have migrated into the right atrium. It was recovered by percutaneous cardiac catheterization through the femoral vein. The patient tolerated the procedure well. The authors believe that this method of retrieval is easier and carries less operative risk than previously reported methods.


Results of retransplantation in individuals who received two successive cadaver kidneys. At the Royal Victoria Hospital in Montreal, 22 patients received two successive cadaver renal transplants. The results were analysed to determine which factors have the best predictive value for success or failure in renal retransplantation. The fate of a second cadaver renal allograft was found to be about the same as the first if the initial transplant has been lost because of rejection and not technical failure. The duration of survival of the initial transplant serves as the best guide to potential outcome of retransplantation after rejection of the initial graft.


Autotransplantation for treating renovascular hypertension. Autotransplantation is presented as a surgical technique for the repair of diseases of the renal artery causing hypertension. Investigation of patients with renovascular hypertension is reviewed stressing the importance of radiologic studies. The author discusses the results obtained in 31 patients who underwent renal autotransplantation. In his experience the procedure has proven to be particularly effective for repairing renal artery stenosis and in patients presenting with severe renovascular hypertension. Associated lesions of the renal artery requiring extensive reconstruction may be repaired with greater safety to both the patient and the kidney using this technique.


Emergency treatment of variceal hemorrhage. The cirrhotic patient with acute bleeding from esophageal varices has less than a 50% chance of leaving the hospital alive; the outlook for survival is so poor that even desperate measures are worthwhile. Some traditional nonsurgical methods for the control of the bleeding are either ineffective at worst or temporary at best. Balloon tamponade is not recommended at all, but intravenously administered vasopressin may be helpful in allowing the necessary diagnostic investigations to be completed. Most important at this stage are the measures necessary to improve the general status of the patient--restoration of blood volume with fresh blood, prevention of ammonia intoxication, support of the liver, correction of metabolic alkalosis and treatment of the hyperdynamic state with digitalis and cardiotonic drugs. Controlling the bleeding is not the greatest problem--the greatest problem is achieving survival of a critically ill patient who undergoes a formidable operation (e.g., variceal ligation stops the bleeding, but is itself an operation of considerable magnitude). In our hands emergency shunting is the best treatment providing a definitive procedure with the highest 10-year survival rate and the lowest complication rate.


[Use of automatic suturing devices in surgery of the digestive tract]. A review of 345 surgical procedures performed on the digestive tract with the use of automatic suturing devices is presented. The morbidity due to hemmorrhage, fistula formation and obstruction as well as the reoperation and mortality rates are discussed and compared with those reported in the literature for the same procedures using standard anastomotic and suturing techniques. This study confirms the safety of these stapling instruments in a wide variety of gastrointestinal procedures in which their use resulted in a considerable saving of operative time.


Occupational lung cancer and smoking: a review in the light of current theories of carcinogenesis. This paper considers modern theories of carcinogenesis as they apply to the induction of lung cancer by tobacco smoking and occupational exposure to carcinogens. Some of the known and postulated factors affecting carcinogenesis are discussed, with particular reference to syncarcinogenesis and thresholds. Factors affecting the intensity of smoking exposure are reviewed, and the generally accepted occupational lung carcinogens are listed. Relative risks for the various carcinogens according to smoking status (where known) are presented. The carcinogens are considered individually, and known or postulated interactions with smoking are discussed. It is concluded that the effects of lung carcinogens can be explained on the basis of current theories that support a rational definition of priorities for the prevention of occupational lung cancer.


Identification of impaired hearing in early childhood. Although the incidence of congenital deafness is high, routine neonatal screening for this problem is not practised, and early identification of congenital or early acquired deafness is relatively rare. Delaying therapy until a child is 3 or more years old severely limits speech development, language acquisition and learning. The commonest causes of delay in diagnosis are the refusal of physicians to listen to the parents' observations, their failure to screen children for hearing and speech problems, and their reluctance to arrange prompt referral for audiologic assessment. Diagnostic delay occurs even though half the children who have impaired hearing are known to be at increased risk. A plea is made for the setting up of a register of infants known to be at risk for impaired hearing. First-contact physicians should be alert to the possibility of hearing problems, particularly in children at high risk. Screening methods for use by nonspecialist practitioners are outlined.


Influence of mechanical ventilation and pulmonary disease on pulmonary artery pressure monitoring. Mechanical ventilation and respiratory disease impose both theoretical and practical limitations on the interpretation of hemodynamic measurements. To properly interpret such information a thorough understanding of the circulatory changes associated with normal breathing, mechanical ventilation and respiratory disease is vital. There are a variety of factors involved in patients with obstructive lung disease and those receiving mechanical ventilation that complicate the usual interpretation of hemodynamic data obtained from flow-directed catheters. An awareness of the potential pitfalls of hemodynamic monitoring in such situations is important in the efficient use of the hemodynamic data obtained.


A phase II trial of ftorafur: adriamycin and mitomycin-C (FAM II) in advanced gastric adenocarcinoma. Fifteen patients with advanced gastric cancer were treated with the combination of Ftorafur, Adriamycin and mitomycin-C (FAM II). Three patients showed partial responses, in five the disease remained stable for at least 3 months and seven showed progression while on treatment. All responding patients showed survival in excess of 12 months. Hematologic toxicity was of only moderate severity. Median white count nadir was 3500 cells/mm3 and median platelet nadir was 187,000 cells/mm3. Four patients had white count nadirs from 2000--2500 cells/mm3 and three had nadirs from 500--1500 cells/mm3; also there were four with platelet nadirs less than 100,000/mm3. However, no drug-related infections occurred and no platelet transfusions were required. The major non-hematologic toxicities of the regimen were nausea, vomiting, dizziness, vertigo, and rhinorrhea. These toxicities were limiting and resulted in termination of the trial because of poor patient acceptance and the failure of the combination to exhibit a therapeutic advantage over the similar combination (FAM) that employed weekly 5-fluorouracil in place of Ftorafur.


5 FU infusion with mitomycin-C vs. 5 FU infusion with methyl-CCNU in the treatment of advanced upper gastrointestinal cancer: a Southwest Oncology Group Study. A randomized trial was conducted by the Southwest Oncology Group (SWOG) in advanced carcinoma of the stomach and pancreas. Patients were assigned to receive monthly 5-fluorouracil 96-hour continuous infusions with either bolus mitomycin-C or oral methyl-CCNU. Mitomycin-C and methyl-CCNU were administered every eight weeks. The 5 FU-mitomycin combination produced a 14% and 22% response rate in disseminated stomach and pancreatic carcinoma, respectively. The combination of infusion 5 FU and methyl-CCNU achieved responses in 9% and 5% of stomach and pancreatic tumors, respectively. There was no significant difference in survival between limbs for either tumor. Median survival in gastric carcinoma on the 5 FU-mitomycin regimen was 25 weeks vs. 18 weeks on the 5 FU-METHYL-CCNU arm. In pancreatic carcinoma median survival on the mitomycin limb was 19 weeks as compared to 17 weeks on the methyl-CCNU program. Leukopenia was greater for the first course on the mitomycin limb. Regression analysis demonstrated that performance status was the most important pretreatment characteristic for predicting survival in both tumors. Neither 5 FU infusion combination appears to significantly alter the dismal prognosis of advanced upper gastrointestinal neoplasms.


The morphologic effects of radiation administered therapeutically for intracranial gliomas: a postmortem study of 25 cases. To investigate the pathologic consequences of therapeutic radiation, this morphologic study evaluated the brains of 25 patients with intracranial gliomas treated both with and without this form of therapy. Although beneficial effects such as the retardation of tumor growth were evident in these studies, among the seventeen patients who received from 5000--6000 rads for malignant gliomas, four developed "late delayed" radiation necrosis. The strong predilection of this tissue response for the white matter adjacent to the neoplasm suggests a local sensitivity which may have been engendered or enhanced by cerebral edema. A fifth case disclosed focal demyelination in the mid-brain suggesting "early delayed" radiation necrosis, and an additional case had distinctive foci of necrosis within the brain stem. Changes of diffuse cerebral edema were noted in many of the radiated brains. It is concluded that radiation therapy in commonly employed dosages for malignant gliomas carries a risk of injury to surrounding cerebral tissues.


Primary carcinoid of the kidney: light and electron microscopic study. A case of an asymptomatic primary renal carcinoid tumor in a 61-year-old male is presented. The tumor was large with extensive hemorrhagic necrosis. Metastases were present in para-aortic lymph nodes. Ultrastructurally, dense core granules and masses of cytoplasmic filaments, including tonofilaments, were seen. The granules were faintly argyrophilic. The relevant literature is reviewed.


Cardiac tamponade resulting from pericardial extramedullary hematopoiesis: a case report and review of the literature. A patient with an unusual myeloproliferative disorder, characterized by hepatosplenomegaly, myelofibrosis, ringed sideroblasts, and conversion to acute myeloblastic leukemia developed cardiac tamponade secondary to pericardia extramedullary hematopoiesis. Diagnostic criteria are discussed. Irradiation and systemic chemotherapy were successful in controlling the effusion.


Clinical toxicologic and pharmacologic studies of gallium nitrate. Gallium nitrate, administered intravenously to patients with advanced cancer, produced renal functional abnormalities consistently at a dose of 750 mg/m2. The toxicity at that dose could be modified by fluid loading and osmotic diuresis. Pharmacokinetic studies revealed a biphasic half-life, (T 1/2 alpha = 87 minutes and T 1/2 beta = 24.5 hours). Osmotic diuresis reduced the urinary concentration of gallium but did not affect serum levels or clearance.


Metastatic melanoma of the gallbladder.

Metastatic melanoma of the gallbladder. Metastatic involvement of the gallbladder in melanoma is rare, but constitutes the most common metastatic lesion involving this organ. Two cases of metastatic melanoma to the gallbladder with radiographic evidence of gallbladder abnormality prior to surgery are presented. These cases are compared to the nine previously reported cases of metastatic melanoma to the gallbladder with abnormal cholecystograms. All eleven cases presented with signs and symptoms compatible with cholecystitis. Nine of the eleven patients had a previous melanoma primary and most had other extrabiliary metastases. Associated cholelithiasis appeared to be only incidental. In addition, nine reported cases of "primary" biliary melanoma were reviewed. Clinical and pathologic presentations in the latter cases were similar to the former cases with metastases. Seventy-eight percent had extrabiliary sites of metastasis at some time in the course of their disease, tending to refute the impression of "primary" biliary melanoma. Melanoma in the gallbladder is much more likely to have metastasized from a regressed skin primary than to have arisen de novo. The two reported cases and the 18 cases from the literature indicate that the physician must consider gallbladder metastasis in melanoma patients presenting with symptoms compatible with cholecystitis.


Multifocal osteosarcoma: a case report with review of the literature. Multifocal osteosarcoma with unique patho-biological features in a 23-year old white male is reported. The natural history and classification of multiple osteosarcomas are reviewed. A proposal is presented for evaluation of the multicentric origin in future cases.


Current concepts of bacteremia in children with malignancies. One hundred fifteen episodes of bacteremia occurred among 2790 children with malignancies hospitalized during a 45-month period. The mean age was 9.3 years with a male predilection (62%). A greater (p less than .025) number of children over 10 years of age died with bacteremia when compared to younger children. The majority of episodes occurred in children with leukemia (56%); however, once bacteremia developed, a significantly (p less than .05) greater number of children with lymphoma died when compared to children with other malignancies. Absolute polymorphonuclear leukocyte counts were greater in survivors (p less than .025) than in children who died. Thirty-seven different microorganisms were isolated with E. coli, S. Aureus, P. aeruginosa, and K. pneumoniae accounting for 50% of the episodes. Anaerobes were isolated from blood of 12 (10%) children. Twelve children had polymicrobial bacteremia and 14 had recurrent bacteremia which occurred during antibiotic therapy. Mortality (78%) in these children was significantly (p less than .001) greater then in children from whom one microorganism was isolated (47%). Interesting aspects include the resurgence of S. aureus, failure of development of meningitis in children with bacteremia, and unchanged antibiotic susceptibility since the last review of bacteremia in this institution. Polymicrobial and recurrent bacteremia necessitate obtaining simultaneous and sequential blood cultures to facilitate administration of appropriate antimicrobial therapy until bone marrow function improves.


Combination chemoimmunotherapy of metastatic breast cancer with 5-fluorouracil, adriamycin, cyclophosphamide, and BCG. One hundred five patients with metastatic breast cancer were treated with 5-fluorouracil, Adriamycin, cyclophosphamide and BCG (FAC-BCG). The results were compared to those observed in a group of 44 patients treated with FAC chemotherapy alone. Although the overall response rates were similar (76% for FAC-BCG and 73% for FAC), the duration of remission was of 9 months for FAC and 14 months for FAC-BCG (p = 0.008). Similarly, survival of responding patients treated with FAC-BCG was significantly longer (24 months) than that observed in the chemotherapy alone treated group (15 months). There was no difference in survival or duration on study for nonresponders. Analysis of response rates by known prognostic factors was unrewarding. The duration of remission and survival, however, were significantly longer for patients with bone soft tissue involvement than for patients with visceral metastasis. Similarly patients with 1 or 2 metastatic sites survived significantly longer than those with more than 3 organ sites involved (p = 0.02). This chemotherapeutic combination is highly effective in inducing remissions. In addition, nonspecific immunotherapy with BCG appears to prolong duration of remission and survival for responding patients.


Mutagenic activity of rhodamine dyes and their impurities as detected by mutation induction in Salmonella and DNA damage in Chinese hamster ovary cells. Commercial rhodamine dyes 6G and B induce His+ reversion mutations in Salmonella and single-strand breaks in Chinese hamster ovary cells, as detected by alkaline sucrose sedimentation. Aroclor 1254-induced rat liver homogenate (S9) is required for production of genetic activity by these dyes. Rhodamine 6G induces both frameshift and base substitution mutations, whereas rhodamine B induces only frameshift mutations. Rhodamine 6G is genetically more active and more toxic than is rhodamine B in both the bacterial and mammalian assays. Rhodamine 6G and B induce doublings of His+ revertants in Salmonella at the doses of 0.02 and 0.52 mumol/plate and shifts in the molecular weight of Chinese hamster ovary DNA at concentrations of 9 x 10(-5) and 9 x 10(-4) M, respectively. All genetic effects assayed demonstrate dose-related increases. Further testing of the pure dyes in Salmonella revealed that rhodamine B loses most of its mutagenicity with purification, whereas rhodamine 6G does not. Impurities from commercial rhodamine B demonstrate the same extent of mutagenicity as the commercial dye.


Survival of patients with metastatic breast cancer treated with either combination or sequential chemotherapy. One hundred twenty-one patients with metastatic adenocarcinoma of the breast were randomized to concurrent combination therapy or single-drug chemotherapy administered sequentially. Although response frequency and duration of response were significantly increased in patients receiving the combination regimen, survival was not significantly prolonged when compared to those receiving sequential treatment. For the 69 patients free of liver metastasis, median survival was comparable in both treatment arms (14.4 months sequential versus 12.8 months combination). These results indicate that a large subset of patients with metastatic breast cancer may benefit from less aggressive therapeutic regimens. Furthermore, these results illustrate that conclusions of chemotherapy trials in breast cancer based only on response frequency and duration of response represent preliminary results subject to change when final survival information becomes available.


Correlation between metabolic reduction rates and electron affinity of nitroheterocycles. Nitroheterocyclic compounds can selectively sensitize hypoxic (tumor) cells to radiation damage in vitro. However, results in vivo have generally been less optimistic, inasmuch as metabolic reduction of these drugs not only limits effective lifetime but also produces metabolic intermediates with marked cytotoxic and carcinogenic activity. With three reducing systems in vitro, E. coli B/r, mouse L-929 cells, and mouse liver microsomes, the rate of nitroreduction of several nitroheterocycles was found to be proportional to their electron affinity (half-wave reduction potential). Relative to the rate of nitrofurazone reduction in each system, metronidazole (Flagyl), N-hydroxyethyl-3,5-dinitropyrrole, misonidazole, nifuroxime, nitrofurantoin, and furylfuramide were metabolized about 200, 20, 2, 1.4, and 1.2 times less rapidly, while 3,5-dinitrobenzonitrile, 2,5-dinitrophenol, and 5-nitro-2-furaldehyde diacetate were reduced 2, 3, and 4 times more rapidly. Since nitroreduction has previously been correlated with subsequent cytotoxicity, DNA damage, and mutagenicity, the present results suggest that improvements in the therapeutic efficacy of nitroheterocycles (i.e., sensitization without toxicity and carcinogenicity) will be dependent on development of drugs with more appropriate pharmacological properties.


In vitro studies of cell-mediated immunity to Moloney murine leukemia virus and Moloney leukemia-associated surface antigens. Cell-mediated immunity to Moloney murine leukemia virus (M-MuLV) and to tumor-associated surface antigens of leukemia cells induced by the virus was studied with an in vitro migration inhibition factor assay. Spleen cells of C57BL/6N mice at Day 14 following inoculation with Moloney murine sarcoma virus, produced migration inhibition factor in response to M-MuLV. The Moloney murine sarcoma virus-immune spleen cells, however, did not respond to other murine type C viruses, to AKR and Rauscher viruses, or to murine mammary tumor virus. The immune spleen cells also responded specifically to purified glycoprotein with molecular weights of 69,000 and 71,000 and proteins with molecular weights of 30,000 and 12,000, but not to protein with a molecular weight of 10,000, of the homologous M-MuLV. Migration inhibition factor production was also observed in response to soluble 3 M KCl extracts of leukemia cells, MBL-2, induced by M-MuLV. Similarly, the immune spleen cells responded to membrane fractions purified from the MBL-2 cells. Comparable membrane fractions prepared from a Gross virus-induced leukemia, E male G2, and a radiation-induced leukemia, RL male 1, were not active. The tumor-associated surface antigens of MBL-2 membranes could be solubilized by the detergent, Nonident P-40. Thus, C57BL/6N mice inoculated with Moloney murine sarcoma virus developed cell-mediated immunity to envelope and some internal antigens of M-MuLV and also to tumor-associated surface antigens of a tumor induced by this leukemia virus.


Mechanism of action of cis-dichlorodiammineplatinum(II). Although the mechanism of action of cis-dichlorodiammineplatinum(II) is not definitely known, much has been discovered regarding its antitumor effect. Although neutral in plasma, the platinum complex becomes aquated intracellularly to form a positively charged platinum species that is likely to interact with nucleophilic sites. The purine and pyrimidine bases of DNA possess such sites and provide the locale for the majority of the biologically effective intracellular platinum binding. Although several types of binding are possible, the biologically important lesions are most likely interstrand and/or intrastrand cross-links. The extent of formation and the rate of cellular repair of these lesions may determine the ultimate effect of cis-dichlorodiammineplatinum(II) on the cell.


Mode of DNA binding of cis-platinum(II) antitumor drugs: a base sequence-dependent mechanism is proposed. Chloroammine and similar complexes of platinum(II) having the ammine ligands in the cis configuration are effective antitumor agents but the corresponding trans isomers are not. This is possibly due to the different manner in which these drugs attack DNA. There is considerable controversy in the literature over the type of DNA lesion caused by cis-platinum(II) complexes. Some have proposed an attack on a single guanine base via chelation to N(7) and O(6) as being the biologically important interaction. However, much indirect evidence suggests that binding to adjacent guanine bases in the same strand of DNA is important to the mechanism of action. Following initial binding to guanine bases, DNA is then locally denatured, exposing additional crosslinking sites. Thus, the selectivity of cis-platinum(II) complexes in inhibiting tumor growth may be due to a combination of intrastrand and interstrand crosslinking to DNA at areas of specific base sequences.


Toxic effects of cis-dichlorodiammineplatinum(II) in man. Administration of cis-dichlorodiammineplatinum(II) may be associated with a number of serious side effects, including nephrotoxicity, gastroeintestinal side effects (nausea, vomiting, and diarrhea), myelosuppression, and occasional transient elevations in liver function tests. In addition, ototoxicity (tinnitus and hearing loss), anaphylactic reactions, peripheral neuropathies, and hypomagnesemia with resulting tetany may also be encountered. The toxic potential of this new agent necessitates careful clinical monitoring during treatment.


cis-Dichlorodiammineplatinum(II) for the treatment of advanced ovarian cancer. cis-Dichlorodiammineplatinum(II) (cis-platinum) was used in the treatment of 25 patients with advanced ovarian adenocarcinoma refractory to alkylating agents. Seven of 24 evaluable patients (29%) responded to cis-platinum (one complete response and six partial responses). Life-table analysis of survival indicates that patients responding to therapy survive longer than those who fail to respond (9 months versus 3.2 months) (P = 0.014). In previously treated patients given 70 mg/m2 iv with forced diuresis, there was substantial nausea and vomiting (100%), hematologic toxicity (67%), and renal toxicity (34%). A review of the single-agent activity of cis-platinum in other ovarian cancer studies indicates an overall response rate of 25% (40 of 161 patients). At least nine combination chemotherapy studies utilizing cis-platinum are in progress, and preliminary information from several of these is encouraging and suggests that cis-platinum-containing combinations may have a major role in the treatment of ovarian adenocarcinoma.


Phase II trials with cis-dichlorodiammineplatinum(II) in the treatment of urothelial cancer. Results of disease-oriented phase II trials with cis-dichlorodiammineplatinum(II) (cis-platinum) in 135 adequately treated patients with advanced urothelial tumors at Memorial Sloan-Kettering Cancer Center are presented. In four protocols which used cis-platinum alone or in combination with Adriamycin and/or cyclophosphamide in 95 patients with bladder cancer, no significant difference (46%--54%) in the number of partial remissions (PRs) in previously untreated patients was noted. The median duration of response in three of the four protocols was 5--7 months. A review of the literature indicates that cis-platinum used singly produced remissions in 45% of 67 patients (95% confidence limit, 12%--57%). In the treatment of superficial bladder tumors, intravesically administered cis-platinum induced few complete or sustained remissions. The difficulties in evaluating response with intravesical therapy are discussed. The importance of patient selection, particularly the need to include patients with objectively measurable disease parameters, in phase II trials is stressed. Differences in patient characteristics and response criteria will necessitate prospective randomized trials of cis-platinum alone versus cis-platinum combination regimens in the treatment of metastatic disease. cis-Platinum was inactive (12% PRs) in 25 patients with prostatic cancer who had objectively measurable parameters. It is of interest that PRs were obtained in three of six patients (50%) with penile cancer. A review of the literature and the data in the present series indicates that cis-platinum has no value in the treatment of metastatic hypernephroma.


Cancer of the ovary: a summary of experience with cis-dichlorodiammineplatinum(II) at the Royal Marsden Hospital. A review of the use of cis-dichlorodiammineplatinum(II) (cis-platinum) as a single agent in 82 patients with advanced ovarian carcinoma, previously treated with chemotherapy, shows that response rates of 33% and 52% are achieved with doses of 30 and 100 mg/m2 respectively. In 58 previously untreated patients a combination of chlorambucil and cis-platinum (regimen B) was compared in a randomized study with a combination of chlorambucil, cis-platinum, and Adriamycin (regimen C). Complete responses were seen in 32% and 41% of the patients respectively. Remissions were most prolonged in patients with complete regressions, the median being greater than 15 months for both regimens. Because of the good regressions, second-look operations have been possible in 12 patients for the purpose of confirming regression and performing radical surgical removal. In six of these patients, all specimens failed to show evidence of residual carcinoma. The major toxic effects of cis-platinum in our hands are neurologic effects and anemia; both have been reversible after cessation of treatment.


Radiotherapy in the management of nonseminomatous testicular carcinoma. Treatment of stage I and II nonseminomatous testicular carcinoma is reviewed. Retrospective studies show comparable cure rates following radiation therapy alone, surgery alone, or radiation therapy and surgery combined. For clinical stage I carcinoma, the long-term disease-free rates appear to be approximately 80%; for clinical stage II disease, the long-term disease-free rates appear to be approximately 60%. The development of effective combination chemotherapy for stage III disease suggests a need to rethink the approach for optimal management of stage II disease. Future clinical trials should be aimed at maximizing cure rates by attention to disseminated disease as well as by local control. Optimal management should also include some consideration of the long-term toxic effects of the various treatment modalities.


Immunocytochemical identification of enkephalinergic neurons in the hypothalamic magnocellular preoptic nucleus of the goldfish, Carassius auratus. Our immunocytochemical investigation of the magnocellular neuroendocrine system in the goldfish hypothalamus reveals enkephalin (ENK)-containing neurons interspersed among the vasotocin (VT)- and isotocin (IT)-containing neurons of the preoptic nucleus. The perikarya of the ENK, VT, and IT neurons do not show distinct morphological differences at the level of light microscopy and are not located preferentially within the nucleus. Separate ENK, VT and IT fibers course laterally and ventrally through the hypothalamus as they descend toward the pituitary gland. All three fiber types form terminals around blood vessels in the neurohypophysis.


The activated hypothalamic magnocellular neurosecretory system and the one neuron--one neurohypophysial hormone concept. The activated hypothalamic magnocellular neurosecretory system of the rat was studied in tissue sections, double stained with the unlabeled antibody peroxidase-antiperoxidase complex (PAP) technique. The results indicate that in animals with an activated hypothalamic magnocellular neuroendocrine system, as well as in normal animals, vasopressin and oxytocin are exclusively synthesized in separate vasopressinergic and oxytocinergic neurons.


The molecular topography of RNA polymerase-promoter interaction. Ultraviolet irradiation forms covalent crosslinks between E. coli RNA polymerase and the lac UV5 promoter substituted with bromouracil in the place of thymine. I have determined the polymerase subunit and the base within the promoter sequence that are joined to each other in two such crosslinks. The sigma and beta subunits of RNA polymerase, respectively, are crosslinked to the third base upstream (-3) and the second base downstream (+3) from the starting point of transcription (+1). Both bases are on the nontemplate strand of the promoter DNA. The location of the beta subunit suggests that it forms at least part of the catalytic site of the enzyme. The disposition of sigma suggests that this subunit plays a direct role in unwinding the DNA at the promoter. The sigma crosslink is close to the "Pribnow Box," which is centered about 10 bases upstream from the RNA start site, contains a striking homology between promoters and is the locus of many promoter mutations.


Isolation of a circular derivative of yeast chromosome III: implications for the mechanism of mating type interconversion. We describe genetic and physical characterization of rearrangements of chromosome III which result in changes of cell type in S. cerevisiae. Two types of rearrangements were obtained as rare events which caused a change at the locus controlling cell type, MAT, associated with a recessive lethal mutation, in one case from MATalpha to MATa-lethal, and in the other case from MATa to MATalpha-lethal. The MATa-lethal mutation is a deletion on the right arm of chromosome III, which we demonstrate extends to (or near) HMalpha. We suggest this deletion removes MATalpha and activates cryptic MATa information stored in HMalpha as proposed in the cassette model of mating type interconversion. The MATalpha-lethal mutation is the result of the formation of a circular chromosome III, which we interpret to remove MATa and activate the cryptic MATalpha information stored at HMa. Strains carrying the MATalpha-lethal chromosome contain a circular chromosome of length 62.6 plus or minus 5.7 mum, which is absent in related strains. This chromosome was confirmed to be chromosome III by hybridization of specific yeast DNA fragments to supercoiled DNA obtained from MATalpha-lethal strains. The isolation of a large circular derivative of chromosome III allows correlation of genetic and physical distance based on large distances-1 centimorgan corresponds to approximately 2700 base pairs.


[Methods of investigating the heart conduction system]. A description is presented of improved methods of examining the conductive system of the heart designed for special routine investigations. The methodology comprises: a) injection of coronary arteries with contrast medium and their X-ray visualization, b) special technique of heart postmortem, c) histological investigation with a view to the precise topography of the particular parts of the conductive system including detailed histological processing in series cestions by means of staining methods: haematoxylin-eosin, elastica-van Gieson, P. A. S., and Heidenhain.


Mutation induction and killing of Escherichia coli by DNA adducts and crosslinks: a photobiological study with 8-methoxypsoralen. Low doses of 350 nm radiation (NUV) in the presence of 8-methoxypsoralen (8-MOP) induce predominantly mono-adducts in bacterial DNA. Further exposure to NUV in the absence of 8-MOP converts a proportion of these mono-adducts to interstrand cross-links. Using this approach the relative effects of adducts and cross-links on bacteria with different repair capacities was studied. Escherichia coli WP100 uvrA recA, believed to be totally deficient in the ability to repair 8-MOP plus NUV damage to DNA, was inactivated on average by a single photon event occurring with a quantum efficiency of about 0.03. We conclude that the inactivating lesion is probably a single mono-adduct. E. coli WP2 uvrA, deficient in excision endonuclease activity, may be inactivated by a very small number of cross-links, probably one. These conclusions are consistent with present knowledge of the repair capabilities of these bacteria. Conversion of mono-adducts to cross-links in WP2 uvrA (which occurs with a quantum efficiency of around 0.3) greatly increases lethality but results in a reduction of the induced mutation frequency presumably because cross-links are (almost) invariably lethal. In the repair-proficient strain WP2 both adducts and cross-links can be repaired but the latter are more likely than the former to lead to either death or mutation.


Biological activity of metal complexes. I. Interaction of pt(II), pd(II) and rh(I) complexes with E. coli strains and with mice LS fibroblasts in vitro. The effects of a number of Pt(II, Pd(II) and Rh(I) complexes against cultures of Escherichia coli (strains B, H10178, uvra-, recA-) and cultures of mice LS Fibroblasts were tested. Most of the compounds showed higher cytotoxic activity than the cis-Pt(NH3)2Cl2, the compound at present on clinical trial as antittumour drug. A new model of active compound is proposed.