Evidence for the multiplication of hepatitis B virus in  "oval cell" culture originated from human embryonic liver. Growth conditions fof human oval cells (immature hepatocytes),evidence of hepatitis B (HB) antigen synthesis in oval cells as revealed by immunofluorescent staining and successful passage of such an agent in the culture fluid up to the 4th passage are described. The results have been proved to be readily reproducible with different inocula. The oval cells used in these experiments were defined as small round cells, with scant cytoplasm, vesicular nuclei and small nucleoli, vitally stained with indocyanine green and synthesizing alpha-foetoprotein but no albumin.


The action of iron on local Klebsiella infection of the skin of the guinea-pig and its relation to the decisive period in primary infective lesions. The infectivity of 16 strains of Klebsiella spp. and its modification by systemic and local ferric iron were tested in the skin of the guinea-pig. The in vivo proliferation of 11 strains was enhanced in varying degrees by Fe+++ (E + strains); 5 strains (Eo) were not enhanceable even by large doses of Fe+++. Of 10 strains examined in detail, 6 were E + and 4 were E0. Guinea-pig and human sera were consistently bacteriostatic for E + strains and bactericidal for Eo strains. Both Fe+++ and microbial iron-chelators abolished the bacteriostasis of E + strains but did not affect the lethal effect on Eo strains. Both effects were diminished by heating the sera to 56 degrees for 30 min and by the anticomplementary substance Liquoid; neither appeared to be due to specific antibody. Virulence, as measured in the skin and by intravenous injection, was roughly associated with degree of enhanceability by iron, the EO strains being among the least virulent. The volume of plasma exudate entering the skin during the first 5 h was sufficient to kill a large proportion of the infecting doses of Eo strains and to inhibit the growth of infecting doses of E + strains. Enhancement of the latter by Fe+++ is predominantly the result of inhibition of the non-specific bacteriostasis exerted by the extravascular plasma. Lesions by E + strains aged 4 h or more are insusceptible to systemic Fe+++ and only moderately susceptible to large doses of local Fe+++. The insusceptibility appears to be due to segregation of the infecting bacilli within exudate leucocytes. Klebsiella infections accordingly provide another example of an initial decisive period of action of the antibacterial defences-in this case non-specific and humoral-which cease to be locally effective after the first few hours. Besides enhancing lesions due to E + strains, systemic Fe+++ has an opposite, apparently anti-inflammatory action on klebsiella lesions, slightly decreasing their size. It was evident with all the strains tested, whether dead or alive, but not in E + lesions in circumstances when they were susceptible to enhancement by the Fe+++.


Immunoglobulin and complement bearing polymorphonuclear leukocytes in allergic contact dermatitis and psoriasis vulgaris. Immunofluorescence techniques were used to demonstrate the presence of immunoglobulin (Ig) and complement (C) bearing polymorphonuclear leukocytes (PMNL) in specimens of skin from the lesions of ten patients with allergic contact dermatitis and of ten with psoriasis vulgaris. The distribution patterns of these Ig- and C-coated cells in the peripheral blood of the twenty patients were also studied. Ten normal healthy controls were included. In cases of allergic contact dermatitis, IgD was the predominant class of Ig on the PMNL. The percentage of C-bearing PMNL were also significantly increased, suggesting that both molecules might be present on the cell memebrane of these cells at the same time. In psoriasis vulgaris, the predominant class of Ig on the PMNL was IgG. The percentage of PMNL coated with C were not signficantly raised. These results suggest that in allergic contact dermatitis the PMNL can be coated with both Ig and C which may render them cytophilic; they also show that in psoriasis vulgaris PMNL can bind Ig. In the ten healthy controls, it was possible to distinguish between those PMNL which are and those which are not bearing immunoglobulin.


Ocular cicatricial pemphigoid associated with practolol therapy. The clinical and laboratory findings in a case of serious cicatricial blistering of the ocular mucous membrane were comparable with those found in cicatricial pemphigoid (ocular mucous membrane pemphigoid).


Secondary skin cancer in nineteenth-century Britain. Clinico-pathological data on secondary skin cancer are of current interest. This paper therefore reviews such data with special reference to nineteenth-century Britain. The old authors were interested in such diverse aspects of the problem of skin secondaries as the mode of invasion, the forms of presentation, the patterns of distribution, clinical diagnosis, microscopic diagnosis, prognosis, and treatment. Although skin secondaries were of serious prognostic import, all cases showing them were not hopeless.


Clinical response of dermatitis herpetiformis skin lesions to a gluten-free diet. Patients with dermatitis herpetiformis have been studied prospectively for 2 years to assess the effect of a gluten-free diet (GFD) on control of the skin lesions. Daily requirements for oral medication with sulphapyridine or dapsone were reduced by GFD treatment and if complete clinical remission of the skin disease occurred, it was maintained while the diet was strictly observed. However, complete remission did not occur significantly more often in GFD-treated patients than in patients taking a normal diet. Many of the latter group exhibited variation in their drug dose requirements during the period of study. GFD treatment seems desirable for the majority of patients with dermatitis herpetiformis, not only to correct the intestinal abnormality but also to minimize the dose of drugs necessary to control the skin lesions.


Ultrasonic methods for sampling human skin micro-organisms. Methods of sampling the microflora of human skin by means of a generator of ultrasonic impulses were compared with the standard washing method in which the skin surface is rubbed manually. The ultrasonic method, at a power setting of 50 W or more, gave essentially similar results to those obtained by the standard method but removed fewer corneocytes.


Melanoma cytoplasmic humoral antibody test. A diagnostic adjunct. Detecting a high titre of antibody directed against the cytoplasm of malignant melanoma cells can help: To diagnose pigmented skin lesions with awkward histopathological signs. The test is positive in about half of the early primary malignant melanomas and in all actively regressing halo naevi. It is negative in benign juvenile melanoma. To prognosticate and provide a continuing assessment of the course of the disease in patients with known malignant melanoma. The test becomes negative when malignant melanoma metastasizes. To alert patients in danger, such as those who have extensive pigmented naevi or have a family history of malignant melanoma, of the supervention of the disease.


Electron microscopy of ancient Egyptian skin. Sections of skin were examined by electron microscopy from the sole of the foot of a 14-year-old Egyptian, who died 3200 years ago and was preserved naturally by desiccation. Remarkable ultrastructural preservation of the epidermal cells and their components was found.


The clinical assessment of topical corticosteroid activity. Glucocorticoid-induced vasoconstriction of human skin has provided a useful guide to topical anti-inflammatory activity. The concentration and base in which a steroid is used influence this activity, and traditional greasy ointments are probably the most effective vehicles. Confirmation of clinical effectiveness should be by double-blind comparison in patients with bilateral skin lesions. Psoriasis provides a more discriminating test of steroidal activity than eczema; the use of occlusive dressings may mask differences between compounds. Short-term trials are informative but in patients with chronic diseases, longer studies should also be done.


The treatment of vitiligo with topical corticosteroids. Light and electronmicroscopic studies. Twenty patients with lesions of vitiligo were treated either with 0.I% betamethasone valerate (BV) or with 0.05% clobetasol propionate (CP) creams and similar control areas with placebo preparations. After 3 months of treatment three of the ten patients treated with BV showed a significant amount of repigmentation only in the steroid-treated areas. Only one of the ten patients treated with CP showed partial repigmentation. Using both light and electron microscopy, histological studies of biopsies taken from control- and steroid-treated areas showed a marked repopulation with functional melanocytes in the repigmenting vitiliginous skin. The melanocytes in these steroid-treated repigmented areas were more dendritic and dopa-positive and, unlike melanocytes in the pigmented margins of untreated areas of vitiligo, contained many melanosomes of normal size, shape and melanization. It appears that both topical corticosteroids are of use for the treatment of selected patients with vitiligo and can induce repigmentation of the skin.


The response of psoriasis to betamethasone valerate and clobetasol propionate. A 6-month controlled study. Twelve patients with symmetrical psoriasis were studied for a period of 6 months. Clobetasol propionate 0-05% ointment was applied to the lesions on one side and betamethasone valerate 0-1% ointment to those on the other side as a double-blind controlled trial of these two treatments. All the patients showed a relative improvement on the side treated with clobetasol propionate. Clobetasol propionate-treated lesions tended to clear for longer than the corresponding betamethasone valerate treated lesions. The condition of five of the patients was better at the end of the study period than at the beginning.


Lymphocyte response to blood transfusion in man: a comparison of different preparations of blood. The appearance of atypical lymphocytes in post-transfusion blood, their incorporation of tritiated thymidine in tissue culture and the elimination of cytotoxic antibody production, have been used as markers to show that frozen red cells are the least immunogenic when compared with dextran sedimented blood and whole blood donations. The absence of atypical lymphocytes and failure to produce lymphocytotoxic antibodies after transfusion of frozen cells is highly significant (P is less than 0.001) when compared with whole blood donations.


Prophylaxis in haemophilia: a double-blind controlled trial. A double-blind controlled trial of prophylactic factor VIII therapy has been carried out on nine severe haemophiliacs at the Lord Mayor Treloar College. Infusions were given once weekly and calculated to give a post-infusion plasma concentration of at least 0.25 I.U./ml of factor VIII. This regime reduced the overall bleeding frequency by 15%. The bleeding frequency in the first 3 days post-infusion was reduced by 66%. A moderate overall reduction in morbidity was also achieved. It is calculated that to reduce the incidence of bleeding in severe haemophiliacs by 15% would require a 73% increased usage of therapeutic materials. More than twice this amount of material is likely to be needed to reduce the bleeding frequency of the same group by 66%.


An evaluation of the luteinizing hormone releasing hormone (LH-RH) test in patients with secondary amenorrhoea. An analysis of the gonadotrophin response to an intravenous injection of LH-RH (50 mug) has been undertaken in 41 patients with secondary amenorrhoea. Thirty-five of the patients were free of any recognizable pathology to account for their amenorrhoea and apparently had a dysfunction of the hypothalamic-pituitary axis. In these patients, the gonadotrophin response to LH-RH was highly variable. There was in general a correlation between baseline plasma LH or FSH levels and their respective increments. There was no correlation, however, between basal oestrogen levels and gonadotrophin increments except in the case of those patients whose basal levels of plasma FSH were higher than those of LH and in those patients whose body weight was less than the ideal for the population. It appears that the gonadotrophin response to a single injection of LH-RH in the majority of patients with secondary amenorrhoea of unknown origin is too variable to be of use either as a diagnostic or prognostic tool.


Radiation-induced immune changes in patients with cancer of the cervix. Immunological methods were used to estimate the therapeutic effect of irradiation and its influence on the immune capacity of patients with cervical cancer. Patients were examined during and following radiotherapy by means of lymphocyte counts, lymphocytic response to phytohaemagglutinin (PHA), and the depressive effect of their sera on the response of normal lymphocytes PHA stimulation. Lymphocyte counts and lymphocyte responses during radiotherapy were depressed to 20 to 50 per cent and 10 to 33 per cent respectively of the values found before treatment. Lymphopenia and depressed T-cell function persisted for years in many patients. The highest values for both lymphocyte counts and reactivities were found in those patients who had unexpectedly good responses to radiotherapy. The immunodepressive effect of patients' sera on normal lymphocyte reactivity decreased and disappeared during therapy and was not present in previously treated patients showing a good prognosis, whereas it showed no change or even increased during therapy in patients with a poor prognosis. The results suggest that not only does immune capacity influence the effect of radiation and the prognosis of the patient, but also that study of the serum-depressive effect could give important information about both the dosage and therapeutic effects of irradiation.


Treatment of Behcet's disease with chlorambucil. Five patients with Behcet's disease in the active and progressive stages were treated with chlorambucil (Leukeran). In all aptients a long-lasting remission was obtained, the ocular lesions did not progress and the vision did not deteriorate. Follow-up varied between 10 and 24 months. Results were encouraging and the form of treatment with chlorambucil and small doses of corticosteroids was superior to other forms of therapy.


Nocardia asteroides keratitis. A case report. An 11-year-old girl developed a corneal ulcer five days after sustaining a corneal abrasion from a fishline sinker. After more than three weeks, the epithelial defect had closed, but superficial stromal opacities in a ring-like distribution had not cleared on topical gentamicin, prednisolone 1% and atropine. Several scrapings disclosed the presence of branching hyphae on smear, eventually identified as Nocardia asteroides by culture. Although sensitivity studies suggested that subconjunctival ampicillin and hourly sodium sulphacetamide would constitute effective treatment, resolution was only partial on this regimen. Only when penicillin G drops were added did the cornea clear maximally and the epithelium heal.


Production of high levels of phosphorylated F1 histone by zinc chloride. Methods have been sought to perturb the level of phosphohistones. ZnCl2 (10 mM) exhibits histone phosphate phosphatase in vivo in HTC cells and leads to hyperphysiological levels of F1 phosphohistone. Treatment of tissue culture cells with this concentration of ZnCl2 leads to a reduction in medium pH to 6.4. Control experiments have indicated that HTC cells grow efficiently at this pH and that the reduction of pH does not produce the hyperphosphorylated state per se. The optimum conditions for the ZnCl2 effect are described. That the effect of ZnCl2 on the heterogeneity of F1 histone is due to an effect on phosphorylation was demonstrated by the observation that the entire effect is abolished by treatment with alkaline phosphatase. The site of phosphorylation is in the carboxy-terminal end of the F1 molecule. The inhibitory effect of ZnCl2 on F3 phosphorylation in metaphase cells is also described.


Peptidoglycan biosynthesis in a thermosensitive division mutant of Escherichia coli. Peptidoglycan biosynthesis during a bacterial division cycle was investigated in the thermosensitive division mutant Escherichia coli PAT 84 Synchronous cell division of this organism was initiated by a shift down from restrictive to permissive growth temperature. Cells harvested at different times after a shift down of temperature served as representatives of the various stages during cell division. These cells were made permeable to peptidoglycan nucleotide-sugar precursors by pretreatment with ether and were found capable of catalyzing the polymerization of externally added precursors as well as the covalent attachment of the newly synthesized peptidoglycan strands to those preexisting in the cell wall. Differences were observed in the rats of peptidoglycan synthesis and in the extent of peptide side-chain corss-linkage at the various stages of division. Nonseptate filaments, formed at the restrictive temperature, incorporated significantly more peptidoglycan which was more cross-linked than in normally dividing cells grown at the permissive temperature. Quantitative analyses of the carboxypeptidase and transpeptidase reactions in cells at different stages of division were performed and the inhibitory effect of a number of beta-lactam antibiotics was investigated. Of special significance was the finding that low doses of penicillin or growth at restrictive temperature, which did not affect transpeptidation, partially inhibited the carboxypeptidase activity. This inhibition was paralleled by an increase in incorporation of newly synthesized peptidoglycan into the preexisting cell wall. We therefore propose that carboxypeptidase activity regulates the availability of peptidoglycan precursor(s) for attachment to the preexisting peptidoglycan by transpepidation.


Unique phenylalanine transfer ribonucleic acids in relaxed control Escherichia coli: genetic origin and some functional properties. Inhibition of protein synthesis in relaxed control E. coli results in the formation of chromatographically unique isoacceptor species of phenylalanine tRNA. The genetic origin and some functional properties of the major unique species of tRNA (Phe) produced during leucine starvation were investigated. RNA:DNA hybridization analyses revealed that the normally occurring and major unique species of tRNA (Phe) are generated from DNA sequences which are identical or closely related and that there may be only one such sequence in the E. coli chromosome. Results from 32P pulse-chase experiments revealed that the unique tRNA (Phe) can be converted to a chromatographically normal form upon resumption of cell growth in fully supplemented medium. These findings, taken with earlier results which indicate that the unique species is not derived from preexisting, normally occurring species, indicate that the unique tRNA(Phe) is a modification-deficient form of the normal species. Comparative studies of the unique and normal phenylalanine tRNAs revealed that the unique species is aminoacylated at a much lower rate than the normal species and is only about 60% as efficient in a tRNA-dependent, poly(U)-directed protein synthesizing system.


Thermal unfolding of yeast glycine transfer RNA. In the present investigations the molecular unfolding of yeast tRNA(Gly) has been studied by a combination of nuclear  magnetic resonance spectroscopy, melting techniques, and relaxation kinetics. From these studies the following pathway of unfolding was found. In a coupled melting transition the tertiary, the DHU, and the anticodon structure are disrupted. This is followed by the melting of the acceptor arm, while the T psi C arm, which only contains G-C pairs, melts out last. Interestingly, during the first melting transition a new structure not belonging to the original cloverleaf structure is formed. The thermodynamic and kinetic parameters of the melting transitions were determined and are discussed in relation to earlier work. The present nuclear magnetic resonance (NMR) experiments as well as earlier studies show that the ring current calculations based on the cloverleaf structure provide a good first-order interpretation of the NMR spectra of tRNA.


A study of secondary and tertiary solution structure of yeast tRNA(Asp) by nuclear magnetic resonance. Assignment of G.U ring NH and hydrogen-bonded base pair proton resonances. The 270-MHz spectra of yeast tRNA(Asp) in H2O solutions containing Mg2+ show clearly resolved resonances in the region from -15 to -9.5 ppm. Resonances between -15 and -11.5 ppm from the hydrogen-bonded protons of the acceptor stem and anticodon arm decrease in intensity with increasing temperature and disappear by 75 degrees C. Simultansously, four well-resolved resonances between -11.2 and -10.3 ppm also decrease in intensity and disappear. Because of this behavior and their positions these resonances have been assigned to the four ring NH protons of G.U base pairs 5 and 30 in the acceptor stem and anticodon arm which are thereby shown not to be hydrogen bonded by normal Watson-Crick hydrogen bonds. The five G.C base pair resonances of the T psi C arm remain visible above 70 degrees C after all other resonances have disappeared. The high-temperature tRNA spectrum agrees well with that of the isolated T psi C hairpin and CCA half-molecule fragments, each of which contains the same five hydrogen-bonded proton resonances. The root-mean-square error between the observed and calculated resonance positions for the hydrogen-bonded base pair protons of these three arms is 0.19 ppm. The dihydrouridine stem is expected to have two A.U Watson-Crick base pairs and no B.C base pairs. However, it does not contribute any hydrogen-bonded resonances to the nuclear magnetic resonance (NMR) spectrum below -11.5 ppm. This suggests that even at 35 degrees C this helix is not hydrogen bonded in a normal manner. In the region below -11.4 ppm there are three additional proton resonances melting earlier than the rest which cannot be assigned to a particular helix of the cloverleaf. We suggest that these resonances arise from hydrogen-bonded protons involved in stabilizing tertiary structure.


Isotope effects and structure-reactivity correlations in the yeast alcohol dehydrogenase reaction. A study of the enzyme-catalyzed oxidation of aromatic alcohols. Steady-state kinetic parameters for the yeast alcohol dehydrogenase catalyzed oxidation of a series of parasubstituted benzyl alcohols-1, 1-h2 and -1, 1-d2 by NAD+ are reported. Catalytic constants have been found to be characterized by large deuterium isotope effects: kH/kD=4.8, p-Br; 4.2, p-Cl; 3, 4, p-H; 4, 2, p-CH3; 3, 2, p-CH3O. The observed isotope effects on k(cat)/K(A), K(A), and K(B), where K(A) and K(B) are Michaelis constants for NAD+ and alcohol, indicate a borderline rapid equilibrium-steady-state kinetic mechanism involving the random addition of substrate and coenzyme to enzyme. With the exception of p-CH3 and possible p-CH3O substituted benzyl alcohol, k(cat) is concluded to represent a single, rate-limiting hydrogen transfer step. A multiple linear regression analysis of the combined data for benzaldehyde reduction (Klinman, J.P. (1972), J. Biol. Chem. 247, 7977-7987, expanded to include p-CH(CH3) 2-substituted benzaldehyde) and benzyl alcohol oxidation has been carried out to determine the contribution of electronic, hydrophobic, and steric effects to k(cat) and substrate binding. Benzaldehyde binding is concluded to depend on electronic substituent effects as previously reported [log 1/K(ald)=(-0.92 +/- 0.18)sigma+-(0.80 +/- 0.067)], whereas benzyl alcohol binding correlates with substrate hydrophobicity [(log 1/K(alc)=(0.60 +/- 0.14) log P -(1.2 +/- 0.12)]. In the case of benzyl alcohol oxidation, k(cat) is independent of electronic and steric effects; the best of seven equations indicates a small negative dependence of k(cat) on hydrophobicity, which is within experimental error or zero [log k(o)=(-0.075 +/- 0.25) log P -(0.65 +/- 0.19)]. Data for benzaldehyde reduction are correlated at the 99% significance level by a single variable equation [(log k(R)=(2.1 +/- 0.37) sigma+-(0.093 +/- 0.14)] and a two variable equation [(log k(R)=(1.9 +/- 0.33) sigma+ + (0.46 +/- 0.20) log P-(0.46 +/- 0.20)]; these equations indicate (a) a large dependence on electronic substituent as reported previously and (b) a possible role for hydrophobic factors in facilitating catalysis. As the result of the observed hydrophobic substituent effects, different ground-state interactions are suggested for the binding of benzaldehydes and benzyl alcohols. Electronic substituent effects lead to the conclusion that there is little or no change in charge at C-1 of substrate at the transition state, relative to alcohol in the ground state. The significance of these effects to the detailed properties of the hydrogen transfer step is discussed.


Freeze etch morphology of outer membrane mutants of Escherichia coli K12. Freeze etching showed that the loss of each of the major outer membrane proteins b, c or d in mutants of Escherichia coki K12 does not influence the morphology of fracture faces of the outer membrane. Mutants that possess a heptose-deficient lipopolysaccharide and which in addition are deficient in one or more major outer membrane proteins exhibit a reduction in the number of intramembranous particles of the outer membrane. Moreover it was shown that lipid phase transitions induce a lateral lipid protein separation in the outer membrane, similar to that found in the cytoplasmic membrane.


Temperature-sensitive formation of the phospholipid molecular species in Escherichia coli membranes. Phospholipids in the membranes of Escherichia coli grown at 37 degrees C are composed of different proportions of molecular species than are those at 17 degrees C. The 1,2-disaturated and 1-saturated-2-unsaturated molecular species increased at 37 degrees C, but the 1,2-diunsaturated species increased at 17 degrees C. When the growth temperature was lowered from 37 to 17 degrees C during the middle exponential growth phase, phosphatidylethanolamine and phosphatidylglycerol composed of proportions of molecular species similar to those found at 17 degrees C were immediately synthesized. By using various membranes composed of different compositions of the phospholipid molecular species, the temperature-sensitive formation of the phospholipid molecular species was found to be independent of the composition of the membrane phospholipids and to be dependent on changes in the specificities of membrane-bound sn-glycerol 3-phosphate acyltransferase against the acyl-CoAs, due to temperature changes.


In vitro repair of radiation-induced strand breaks in DNA. DNA, gamma-irradiated in vitro or in isolated thymocytes was treated with several enzymes to achieve repair of the radiation-induced single strand braks. Whereas an incubation with polynucleotide ligase can join only 25% of the single strand breaks, a combined treatemnt with exonuclease III (EC 3.1.4.1), DNA polymerase I (EC 2.7.7.7), and polynucleotide ligase leads to repair of 80% of the breaks. For this in vitro repair the exonuclease III has to remove several, probably damaged, nucleotides from the 3'-terminal producing a single-stranded gap, which will be filled in by DNA polymerase I and joined by ligase. Tests for successful rejoining of the strand breaks were performed by showing the loss of primer 3'-OH sites for DNA polymerase I, by the resistance of incorporated nucleotides in the gap to removal by a second exonuclease III treatment, and by strand break determination in the analytical ultracentrifuge. 20% of the radiation-induced strand breaks will not be repaired by this combined treatment possibly due to an incomplete binding of the ligase on the 5'-terminals and/or an incomplete removal of the damaged 3'-terminals by exonuclease III.


Specific cleavages of pure tRNAs by plumbous ions. After renaturation some pure tRNAs were submitted to the action of lead acetate 1 . 10(-3) M at pH 7.3 and at 37 degrees C in the presence of either 1 M or 0.5 M NaCl. These tRNAs were specifically cleaved by Pb2+. The exact cleavage points were determined by analysing the oligonucleotides obtained from three yeast tRNAs. In 1 M NaCl, tRNA(Phe) is cleaved after the hUp17 and partially cleaved after Cp73. In 0.5 M NaCl, there are cleaveages after hUp16, hUp17 as well as a partial one after pGP1. In 1 M NaCl tRNA(Asp) is not cleaved, whereas in 0.5 M NaCl 50% of the molecules are cleaved in the anticodon region after Up35, 14% after hUp19 and 6% after hUp16. In 1 M NaCl tRNA(Val) is cleaved in the hU loop: 40% after hUp16 and 60% after CP17. The action of lead on five other pure tRNAs was studied on the analytical scale only, by polyacrylamide gel electrophoresis. They could be classified into two familites, one cleaved mainly in the hU loop, the other in the anticodon loop. The minimal concentrations of Pb2+ required for cleavage were determined for several tRNAs, the most sensitive of which, yeast tRNA(Val), being still cleaved with a concentration of 5 . 10(-6) M in 0.15 M NaCl. Although the cleavage often occurs after hUp, poly (hU) is less sensitive than poly(U). This and other results indicate that cleavages depend more on the conformation than the sequence of the polynucleotide chain, bends in the tertiary structure being lead-sensitive sites. Finally, the amino acid acceptance activities of cleaved tRNA(Phe) and tRNA(Asp) were determined.


The binding of lac repressor and the catabolite gene activator protein to halogen-substituted analogues of poly[d(A-T)]. We have measured the binding of two regulatory proteins to the complete halogen-substituted series of poly[d(A-T)] analogues. Both the lac repressor and the catabolite gene activator protein were found to bind more strongly to all of the halogen-substituted DNAs than they do to poly[d(A-T)]. For both proteins, the order of binding preference is poly[d(A-ioU)] is greater than poly[d(A-brU)] is greater than or equal to poly[d(A-clU)] is greater than poly[d(A-flU)] is greater than poly[d(A-T)]. Quantitative data on the binding of these proteins to poly[d(A-U)] is also given. The significance of these results for the mechanism of protein-DNA interaction is discussed. This is the first report that an activator protein binds more strongly to a halogen-substituted DNA; we discuss this result with regard to the mechanism of action of bromodeoxyruidine and other halogen-substituted base analogues on the inhibition and induction of differentiation in eucaryotic cells.


The effect of thioketo substitution on uracil-2-aminopurine and uracil-2, 6-diaminopurine interactions in polynucleotides. The existence of the complexes poly[r(s4U)] . poly[r(n2h6A] and poly-[r(s4U)] . poly[r(n2A)] was demonstrated by means of spectrophotometric titration and sedimentation veolicty analysis. According to the absorption-temperature profiles thioketo substitution of poly[r(U)] . poly[r(n2h6A)] led to stabilisation of the helical structure, thus implying that the 4-thioketo group does not participate in s4U . n2h6A base pairing. In the case of poly[r(s4U)] . poly[r(n2A)] drastic destabilistaion of the helical structure by thioketo substitution was observed. This indicates that the thioketo substituents participate in s4U .n2A base pairing.


The role of energy-generating processes in the degradation of guanosine tetrophosphate, ppGpp, in Escherichia coli. The effect of energy limitation on the intracellular level of guanosine 5'-diphosphate, 3'-diphosphate (ppGpp) was investigated. Glucose exhaustion, as well as uncoupling of oxidative phosphorylation, evokes an immediate increase in the ppGpp concentration. This increase is a consequence of a lowered degradation rate of the nucleotide. The level of ppGpp, which determines the rate of ribosomal RNA synthesis, is thus subject to a double control system: amino acyl-tRNA availability governs the rate of synthesis while energy metabolism governs the rate of degradation.


Peptides isolated from Enterobacter nuclease as potential polyamine binding sites. The Enterobacter nuclease, which cleaves RNA between the 3'-phosphate group of cytidylic acid and the 5'-hydroxyl group of adenylic acid, has been shown to be affected by the polyamines, spermidine, spermine and putrescine. These substances enhance the hydrolytic activity of the enzyme against both poly(C) and yeast RNA. Sperimidine and spermine also reverse the inhibition of the enzyme by the ordered polynucleotides, apparently by removing them from the surface of the enzyme. Treatment of poly(G)-bound peptides (obtained from tryptic digests of poly(G)-bound nuclease) with an excess of spermidine resulted in the isolation of spermidine-bound peptides. Purification of these peptides through ion-exchange chromatography resulted in the isolation of three spermidine-bound peptides which consisted of 17 residues (6 amino acids), 19 residues (10 amino acids), and 12 residues (9 amino acids). The binding ratio of spermidine to peptides varied from 1:1 to 3:1.


The similarities of ribosomal and basic chromosomal proteins from fungi. We have compared the physical and chemical properties of yeast and fungal ribosomal proteins with those of higher eukaryotic histones. We have found that acidic urea gel electrophoresis, sodium dodecyl sulfate gel electrophoresis or chromatography on carboxymethylcellulose columns failed to distinguish ribosomal proteins from histones. The majority of the ribosomal proteins did not adsorb to an amberlite CG-50 column in the presence of 8% guanidine hydrochloride. Histones quantitatively adsorbed to an amberlite CG-50 column in the presence of 8% guanidine hydrochloride. A small number of fungal acid-soluble nuclear proteins, which coelectrophoresed with histones, were identified in a presumed nucleolar and nuclear membrane fraction. This fraction contained large amounts of RNA and small amounts of DNA. It is suggested that contamination of yeast and fungal chromosome preparations by a small number of ribosomal proteins can occur. Furthermore, several commonly employed criteria did not distinguish contaminating ribosomal proteins from authentic histones.


Transplantation versus prosthetic replacement of the heart. The cardiac allograft has a far superior clinical record than the mechanical device or artificial heart. Further efforts in two-staged cardiac replacement are encouraged using, initially, an orthotopic total replacement of the nonfunctioning heart and, later, substitution by a cardiac allograft. Orthotopic cardiac replacement with a mechanical device offers promise, but elective clinical application may be delayed for many years.


Human tumour xenografts.

Human tumour xenografts. The historical development, and more recent advances in the use of xenografts of human tumours as potential models for the study of human cancer are described and discussed. The practicality of the various systems available are critically reviewed, as well as their potential application in the field of experimental chemotherapy. Emphasis is given to the fact that over two decades of research effort has hardly produced any results of clinical relevance, often because workers have been too carried away with their technical achievements to appreciate the rules of experimental chemotherapy. The role of human tumour xenografts in the study of human cancer is appreciated, but the development of newer syngeneic, differentiated animal models may lessen their importance in the field of experimental chemotherapy.


"Xenogeneic resistance" to rat bone marrow transplantation. III. Maturation age, and abrogation with cyclophosphamide, Corynebacterium parvum and fractionated irradiation. Lethally irradiated C57 Bl/6 mice and (C57 X A) F1 hybrids fail to accept doses of rat bone marrow cells (5 X 10(6)) which give confluent splenic repopulation in "non-resistant" strains of mice. This phenomenon has been termed "xenogeneic resistance" (XR). XR in (C57 X A) F1 mice can be overridden by a very large inoculum of rat bone marrow (26 X 10(6) cells). XR is not manifest in mice of a resistant strain at ages of 18 days or younger, but is manifest at ages of 22 days and older. XR can be abrogated by agents as varied as: 1) cyclophosphamide, which abrogates XR in a dose dependent manner when given 1 hr prior to lethal irradiation and bone marrow transplantation 2) C. parvum, which abrogates resistance when given 7 days prior to lethal irradiation and bone marrow transplantation, and 3) Fractionated irradiation, which, while capable of abrogating XR, is much less potent than either cyclophosphamide or C. parvum.


Inhibition of transplanted sarcomas mediated by BCG in rats with a defined immunological deficit. Experiments were undertaken to test the hypothesis that a major component of BCG contact-induced inhibition of syngeneic tumour growth in rats is not dependent on the participation of thymus-processed (T) cells. Hosts were deprived of T cells by thymectomy followed by either lethal irradiation (850 rad) and bone marrow reconstitution, or repeated whole body irradiation to a total dose of 1,000 rad. After 6 weeks had elapsed to allow for bone marrow restitution, rats were challenged with trypsinized sarcoma cells admixed with Glaxo strain BCG. For sarcoma P7, host T-cell deprivation did not significantly diminish the capacity of BCG to prevent the progressive development of this neoplasm from an inoculum of one million cells. Under similar conditions, the incidence of sarcoma CC5 development in maximally deprived hosts was significantly greater (7/19) than in normal controls (1/16) (P is less than 0.05), but the majority of rats (58%) did not succomb to tumour outgrowth. In the case of a third neoplasm--a spontaneously metastasizing fibrosarcoma (P8)--the effect of BCG on primary tumour development was comparable in normal and deprived recipients and was limited to temporary arrest as distinct from complete inhibition. Assessment of the influence of BCG on lung metastases was more complex since the extent of metastatic disease from subcutaneous tumour cells alone was greater in deprived rats than in normal rats. It is concluded that T-cell participation is not a major requirement for BCG contact-induced inhibition in this system and some implications for the mechanism of action are discussed.


[Pharmacokinetic characteristics of a massive dose of furosemide in acute kidney failure]. A pharmacokinetic study with high doses of frusemide has been performed in nine patients with acute tubulopathy, treated by hemodialysis. The patients were studied between 6 and 28 days after the beginning of the disease. Renal function was normal before the episode of acute renal shutdown. The diuretic effect of frusemide was closely related to creatinine clearance (Ccr): no effect in anuric cases, moderate effect if Ccr was lower than 10 ml/mn, marked effect if Ccr was higher than 10 ml/mn. no direct relationship could be demonstrated between drug plasma level and natriuresis. Frusemide half-life in plasma was short even in anuric patients, suggesting the existence of an extra renal excretion and/or inactivation pathway. Hence toxic side effects due to drug accumulation should not appear in the recommended 1 g/day regimen.


Influence of salts and gelatin on disintegration of Saccharomyces cerevisiae by freeze-pressing. The disintegration by freeze-pressing of a low concentration of Saccharomyces cerevisiae suspended in aqueous solutions of gelatin and different salts has been studied at different temperatures. In the freeze-pressing process deionized water and salt solutions flow in pulses, whereas samples with increasing concentrations of gelatin or cells tend to flow more smoothly. This smooth flow enhances the disruption efficiency particularly at lower temperatures, which seems to be of great practical importance. The addition of salts also promotes disintegration. The presence of both gelatin and salts works antagonistically on disintegration presumably because of different modes of action at disruption of cells.


Cell inactivation by ultrasound.

Cell inactivation by ultrasound. Cell survival curves have been obtained for Escherichia coli B (E. coli B) after the sonication of suspensions of the bacteria with continuous wave ultrasound at a fixed frequency of 2 MHZ between peak intensities of 8.7 and 2.25 W cm-2. It was found that under suitable conditions the survival curves were reproducible and it also was found that there was a clear relationship between the rate of inactivation and the peak acoustic intensity of the ultrasound. There appeared to be a lower threshold of peak intensity below which no inactivation was observed.


Dynamics of mixed cultures of Lactobacillus plantarum and Propionibacterium shermanii. The mixed culture of Lactobacillus plantarum and Propionibacterium shermanii grown anaerobically in glucose minimal medium exhibits features typical of a commensal interaction even though a number of complicating factors, such as a large maintenance requirement of L. plantarum and inhibition of growth of P. shermanii at low pH, are present. A simple mathematical model of the system is presented and is shown to reproduce rather well some of the features of the continuous mixed culture system in both steady-state and transient situations.


The kinetics of protein salting-out: precipitation of yeast enzymes by ammonium sulfate. Protein solubility can be adequately represented by the classical Cohn equation for the salting-out of alcohol dehydrogenase and fumarase from clarified yeast homogenate with ammonium sulfate. However, the constant beta in this equation is a function of the contacting procedure employed. The kinetics of continuous salting-out were similar for alcohol dehydrogenase and fumarase. The overall rate equation for precipitation had a variable order which was high initially, up to 3.1, but approached unity on completion of precipitation. This was followed by a partial resolution stage which was first order with respect to the concentration driving force. Precipitate particle size was estimated as 0.5 to 5 mum with continuous flow precipitation producing the largest particles.


Yield and maintenance relations of yeast growth in the chemostat at superoptimal temperatures. A model is proposed that accounts for the decreases in yield which occur in chemostat cultures of mesophilic yeasts at superoptimal growth temperatures. Two yield depressing effects were identified, one due to increased maintenance requirements by the viable fraction of the population, the other due to energy substrate dissipation by the nonviable function. The two effects are functions of the dilution rate, as is the fraction of nonviable cells. Experimental results were obtained on the yield, maintenance, and dissipation of energy substrate in a glucose-limited chemostat culture of a respiration-deficient mutant of Saccharomyces cerevisiae at 39 degrees C. The rates of glucose utilization for maintenance and for dissipation constituted, respectively, 33-28% and 15-9% of the total glucose utilization rate over the range of dilution rates tested (0.038-0.064 hr-1), while the yield varied over this range from 0.066-0.085 g of biomass (dry wt) per gram of glucose.


Pilot scale exponential growth of Escherichia coli W to high cell concentration with temperature variation. An efficient method to grow Escherichia coli W to high cell concentrations on the pilot scale is described and discussed. The method involves growth linked introduction of glucose and ammonia to the culture, sparging with oxygen, and maintenance of aerobic conditions by gradually decreasing the temperature in the culture in order to keep the oxygen demand within the limits of the capacity of supply. Under these conditions the linear rate of cell mass production is actually the result of exponential growth with a gradually decreasing growth-rate constant. About 10 kg packed cells were produced in a 50 liter working-volume fermentor in one run of 13 hr. The concentration of the cells at the end of the growth was about 47 g dry cells/liter. The expenditure for nutrients was minimal and the controls were of simple automatic nature. From the determined yield constants for glucose, nitrogen, phosphorus, and oxygen it may be inferred that the cells grown by this method are similar to those grown exponentially at constant temperature.


Optimal control of a semibatch fermentation. This work is concerned with the optimization study of the semibatch fermentation by which an amino acid is produced. The particular fermentation studied is the synthesis of lysine by the auxotrophic mutant. Applying Green's theorem to the maximization problem was proposed, and it succeeded in determining the feed rate of the substrate that maximized the production rate of the desired product.


Influence of cell concentration, temperature, and press performance on flow characteristics and disintegration in the freeze-pressing of Saccharomyces cerevisiae with the X-press. The pressure required for initiation of flow when freeze-pressing with the X-press is related to the phase boundaries of water, particularly those between ice I and liquid even at temperatures around -25 degrees C and lower. Widening the orifice of the pressure chamber to diameters larger than 2.5 mm leads to lower pressures and less extensive cell disintegration. Pressing Saccharomyces cerevisiae slowly with the aid of a manual hydraulic jack at -25 degrees C produces a disintegration of 60-75% irrespective of cell concentration. Pressing at -35 degrees C shows no clear differences. Pressing more rapidly with the aid of a motor-driven hydraulic press produces a similar extent of disruption of diluted cell suspensions (5.4 mg/g) as slow pressing. However, freeze-pressing a paste of baker's yeast (270 mg/g) increases the degree of disintegration. Under these conditions the disintegration is further enhanced by a lower temperature, -35 degrees C, and by a high velocity of flow through the orifice, such that more than 95% of the S. cerevisiae is disrupted by one pressing at less than 2 X 10(8) Pa. Mechanisms for flow through the X-press are suggested and discussed in relation to the phase diagram of water.


[Elimination of allogeneic inhibition and change in the differentiation of mouse bone marrow colony forming cells exposed to syngeneic lymphocytes]. A mixture of the bone marrow cells and lymphocytes of the lymph nodes of C57BL mice was injected intravenously to A X C57BL mice irradiated in a dose of 850 R. It appeared that under the effect of syngenous lymphocytes of C57BL mice the colony-forming cells (CFC) of the bone marrow of the F1 hybrid mice proliferated as if they were in the syngenous organisms. Along with the restoration of the number of CFC capable of forming colonies in a genetically foreign organism these cells, under the effect of lymphocytes, altered the direction of differentiation which was characterized by increase in the number of granulocytic colonies. This process was accompanied by the development of the blast transformation reaction and hyperplasia of the lymphoid tissue of the recipient spleen.


[Development of immune tolerance to microbial and cross-reacting tissue antigens in mice]. C. albicans vaccine was administered to newborn mice (CC57BR). A tolerance to the microbial antigen was thus obtained which depressed the reaction of rejection of the C3H mice skin allograft. Tolerant mice displayed a reduction of the number of lymphoid cells reacting with the microbial and tissue antigen of the donor in the reactions of immunoadhesion and blasttransformation; there was also seen a reduction of the cytotoxic activity to the fibroblasts of C3H mice. The tolerance to the microbial and cross reacting transplantation antigens was pased by the spleen cells to syngenous recipients irradiated in a dose of 700 R.


Sex difference and the influence of endotoxin and hypertransfusion in spleen colony formation. Lethally whole-body irradiated mice upon receiving a relatively small quanity of marrow cells show a distinct sex difference in ability to support colony formation in spleen and cell proliferation in femoral marrow. The difference should be defined with reference to the organ in question. Female mice manifest more efficient splenic haematopoiesis while among the males myeloid haematopoiesis, as measured by 59Fe and 125IUdR incorporation, seems to be greater. Endotoxin treatment and hypertransfusion exert profound influences, interacting with the sex-hormonal status of the animal.


The siting of comparable skin flaps in pigs. The mean surviving length of flaps of the same size on the pig's abdomen was measured; there was a statistically significant difference in the surviving length of flaps based side by side, whereas the difference in surviving lengths of flaps based opposite to each other was not significant. It is suggested that flaps used for comparison in experiments on pigs should be based opposite each other and not adjacent.


Proximal and distal releasing incisions for the treatment of flexion contracture of the popliteal region. Proximal and distal releasing incisions and skin grafting have been found to be useful as an alternative procedure to the standard transverse incision for dealing with flexion contractures of the popliteal region. In this region important structures underlying the deep fascia may readily be exposed when the deep fascia itself is involved in the scar process and is cut across during the release. Chronic ulcer formation in the centre of hypertrophic scar contracture is a fairly frequent accompanying feature. Proximal and distal release incisions help solve the problem by providing protection for the deep vessels, nerves and tendons, without jeopardising the take of a skin graft.


Hypnosis in pedicle graft surgery.

Hypnosis in pedicle graft surgery. The use of hypnotherapy to modify the reactions of 13 patients undergoing pedicle flap operations is described. The morale and mental approach of these patients was improved. Premedication and hypnotic drug requirements were reduced to nil following hypnotic training. Analgesic medication was also reduced, not being required at all following nearly two-thirds of the operations.


Ileal urinary diversion in patients with Renal transplants. Ileal urinary diversion has been carried out on 8 occasions in 7/215 kidney transplant patients. In six instances the diversion was required either because the bladder had previously been removed or there was gross abnormality of the outflow tract: in 2 it was successfully employed to deal with difficult post-transplantation urinary fistulae. 4 patients died: 3 within 2 months and 1 at 9 months after the operation. Infection with organisms indistinguishable from those cultured from the ileal loop, was a common complication and, although there were other significant factors which contributed to the mortality, retrospective consideration suggests that energetic prophylactic antibacterial therapy should be instituted when kidney transplantation with ileal urinary diversion is undertaken.


A comparison between suction and corrugated drainage after simple mastectomy: a report of a controlled trial. A randomized prospective trial of closed suction wound drainage compared with corrugated wound drainage after simple mastectomy for early breast cancer (lesion confined to the breast and without skin ulceration) is reported. Fifty-one patients were admitted to the study and there was no difference between the two groups with respect to local complications such as ifection, skin necrosis and collection under flaps and wound healing. Dressings require to be performed significantly less frequently with suction as opposed to corrugated drainage and suction drains were removed significantly sooner than corrugated drains. It is concluded that closed suction wound drainage should be used after simple mastectomy.


Surgical research in the United Kingdom: past, present and future. The history of surgical research in the United Kingdom leading to the foundation of the Surgical Research Society (SRS) is briefly reviewed. The tendency of the SRS has been to become increasingly rigorous, reductionist and a forum for work which is not only a high standard but also largely completed. This has had both good and potentially bad effects. Surgeons should realize that their major involvements in research will lie in the relatively untidy field of clinical science, and it is hoped that this view will continue to influence the activities of the SRS.


Benign obstruction of subclavian and axillary veins possibly due to filariasis. This paper analyses the clinical features of 8 patients with post-thrombotic obstruction of the subclavian and axillary veins demonstrated by phlebography. The classic physical signs of swelling of the arm with dilated superficial collateral veins were seen. The causes of thrombotic obstruction are discussed and the precipitating cause in this study was possibly filariasis.


A controlled trial of intraperitoneal noxytiolin in perforated appendicitis. A controlled trial of intraperitoneal noxytiolin (Noxiflex) solution in patients with peritonitis due to perforated appendicitis is reported. No significant difference was found in the incidence of complications between the treated and a control group, raising doubts as to whether this substance is of value when used intraperitoneally in peritonitis.


Trial of combination of guanethidine and oxprenolol in hypertension. Thirteen hypertensive patients entered a double-blind crossover trial of guanethidine and oxprenolol in combination. In nine patients who completed the trial there was an additive effect on blood pressure, but the combination had a smaller effect on heart rate than was expected from the individual effects, and side effects were not increased. During treatment with oxprenolol the plasma potassium concentration rose from 3.6 mmol (mEq)/1 to 3.9 mmol (mEq)/1. No correlation was found between the plasma oxprenolol concentration and changes in blood pressure or response to injected isoprenaline, but measurements of plasma oxprenolol concentrations were of value in determining compliance with the protocol.


Bran tablets and diverticular disease.

Bran tablets and diverticular disease. Three treatments for patients with symptomatic diverticular disease were compared in a crossover trial. Neither a high-roughage diet (HRD) nor Normacol plus an antispasmodic were as effective as bran tablets, which produced a significant increase in daily stool weight and a decrease in the intestinal transit time. An abnormal rapid electrical rhythm in colonic smooth muscle was found initially in 80% of the patients, but the incidence was reduced by each treatment after one month; it was present in only 40% of patients after treatment with bran tablets. Only bran significantly reduced a high percentage motility to within normal limits. Bran proved to be the most effective treatment, not only in improving the symptoms in patients with diverticular disease but also in returning to normal the abnormal pathophysiological changes. Bran tablets were both convenient and acceptable as well as effective.


Once-daily dosing with Atenolol in patients with mild or moderate hypertension. Because of the difficulties patients have in adhering to their drug regimens a trial was performed in which patients with essential hypertension were given, in random order and for four weeks each, three different doses of atenolol to be taken once daily. Atenolol effectively decreased lying and standing blood pressures, and there was no difference between the effects of the three doses. The simplicity of the regimen, as well as atenolol's freedom from troublesome side effects, should be valuable in helping patients adhere to long-term treatment.


Prevention of postoperative deep vein thrombosis with dipyridamole and aspirin. One hundred and sixty patients have been studied in a controlled trial of two drugs which have been in combination to prevent postoperative-deep-vein thrombosis. Both the drugs, dipyridamole (Persantin) and aspirin, were given by mouth from the evening before operation and for seven days after operation. The radioactive fibrinogen test was used to diagnose thrombosis. Of the 85 patients in the control group 24 developed thrombosis (28%). Twelve out of 85 patients (14%) in the test group were found to have thrombosis. Similar significant differences in incidence were found in the various subgroups.


Management of early cancer of the breast. Report on an international multicentre trial supported by the Cancer Research Campaign. In the largest controlled clinical study so far undertaken on the management of early cancer of the breast in women the results of a radical regimen (simple mastectomy and radiotherapy) were compared with those of a conservative policy (simple mastectomy alone). The study included enough patients (2268) to ensure that small but significant differences between the two treatments would emerge. With a five-year follow-up there was no evidence that routine postoperative radiotherapy was harmful or that it conferred further benefit as regards survival or distant recurrence. Radiotherapy did, however, reduce the incidence of local recurrence. Neither form of treatment can counter occult disease far from the breast that is present at the time of initial treatment, which probably accounts for the high death rate from breast cancer, but the results of the study would support the view that conservative primary treatment and subsequent adjuvant chemotherapy may be the treatment of choice in future.


Combined alpha- and beta-adrenoceptor blockade with labetalol in hypertension. The antihypertensive effect of labetalol, a new alpha- and beta-adrenoceptor inhibiting agent, was studied in 20 patients in a double-blind crossover trial. A dose of 300 mg daily reduced blood pressure only moderately in the supine position, though in the sitting and standing positions the effect was more pronounced. A dose of 600 mg daily produced statistically significant and clinically relevant reductions in blood pressure in all positions studied. The effect on heart rate was small and of significance only in reducing the heart rate increment due to a change in posture. Side effects were mild: only one patient complained of postural dizziness with the higher dose. We conclude that labetalol is useful in the treatment of mild and moderately severe hypertension.


Four new anti-inflammatory drugs: responses and variations. Ninety patients with rheumatoid arthritis completed a double-blind crossover trial comparing fenoprofen, ibuprofen, ketoprofen, and naproxen. Fenoprofen and naproxen were slightly more effective than the other two drugs but there were striking individual variations in response. Groups of patients could be identified who preferred each of the four drugs. The commonest side effects were those related to the upper gastrointestinal tract; these showed individual variation and seldom occurred with more than one or two of the drugs. Side effects were least common with ibuprofen and naproxen. Since naproxen combined greater effectiveness with a lower incidence of side effects it must be regarded as the first choice among these drugs. It may be necessary to try several drugs before finding the right one for a particular patient.


Azathioprine and penicillamine in treatment of rheumatoid arthritis: a controlled trial. Sixty-five patients, 33 receiving azathioprine and 32 receiving penicillamine, took part in a one-year, single-blind external-observer trial designed to compare the efficacy and toxicity of these two drugs in the treatment of rheumatoid arthritis. By six months there was a significantly greater rise in haemoglobin and fall in erythrocyte sedimentation rate among those receiving penicillamine, and by one year this difference remained only in the increase in haemoglobin levels. Fifteen patients, 10 on azathioprine and 5 on penicillamine, had to stop treatment because of side effects; 90 single side effects occurred, 48 in those on penicillamine and 42 in those on azathioprine. After one year both drugs were similar in efficacy and toxicity, but longer-term trials are needed. Both drugs were effective.


A controlled comparison of flupenthixol and amitriptyline in depressed outpatients. Sixty depressed outpatients were allocated to treatment with either amitriptyline (75-225 mg/day) or flupenthixol (1-5-4-5 mg/day) in flexible dosage for six weeks under double-blind procedures. Various objective and subjective assessments were carried out before and after one, three, and six weeks of treatment. Twenty-three patients completed the course of amitriptyline and 28 the course of flupenthixol. Almost all variables improved significantly over time, irrespective of drug. On most ratings there were no significant differences between the two drugs, but the trends favoured flupenthixol. In particular, flupenthixol lessened anxiety scores more than amitriptyline. Unwanted effects were few and not troublesome except in two patients receiving amitriptyline. Flupenthixol, in low dosage, is a useful alternative antidepressant for depressed outpatients.


Controlled trial of oxygen in uncomplicated myocardial infarction. Two-hundred consecutive patients thought to have suffered a myocardial infarction were admitted to a randomised, double-blind controlled trial of oxygen or air administered by MC mask throughout the first 24 hours in hospital. Forty-three patients in whom myocardial infarction was not subsequently confirmed were excluded from the analysis. The remaining air and oxygen groups were comparable except for a significantly higher PaO2 and serum aspartate aminotransferase level in the oxygen group. There was no significant difference in mortality, incidence of arrhythmias, use of analgesics, or systolic time intervals between the two groups, although a higher incidence of sinus tachycardia was found in those given oxygen. There appears to be no evidence of benefit from the routine administration of oxygen in uncomplicated myocardial infarction.


Bacterial glutaminase in treatment of acute leukaemia. A glutaminase-asparaginase enzyme from Achromobacter sp has antitumour activity in vitro and in animals. Glutaminase was administered in doses of 3500-20 000 IU/m2 body surface area/day to six patients with acute lymphoblastic leukaemia (ALL) and three patients with acute myeloid leukaemia (AML). The enzyme had a blood half life of 80 minutes but depletion of blood glutamine persisted for 12 hours after single doses. Seven patients, including four (two with AML and two with ALL) resistant to asparaginase, received repeated doses of glutaminase. Antileukaemic effects were observed in all seven; one elderly patient developed metabolic acidosis. Study of this new antileukaemic agent in patients with acute leukaemia at an earlier stage of their disease is now justified.


Functional landscapes of the dominant hemisphere. There is abundant evidence that in non-anoxic brain tissue the blood flow is controlled mainly by the functional activity of the neurons. This enables the use of regional blood flow measurements for the localization and quantitation of events in the human brain which correlate to normal and abnormal mental activity. Measurements of regional cerebral blood flow (rCBF) by means of the 133Xe clearance technique have been demonstrated that the activity distribution in the normal resting brain shows a typical "landscape", with the highest activities in the premotor and frontal regions and the lowest in temporal and parietal ones. The resting pattern is readily changed by voluntary motor activity which gives a rolandic-postrolandic activity peak and by sensory stimulation which activates rolandic and prerolandic structures especially. Speech and reading activates the upper, anterior and posterior speech cortices, as well as the face, tongue and mouth area. Problem solving also activates premotor and frontal regions especially and, if visual activity is involved, the occipital pole is also activated. These normal patterns confirm by and large well known aspects of the functional anatomy of the human dominant hemisphere.


Blood-brain barrier studies in the rat: an indicator dilution technique with tracer sodium as an internal standard for estimation of extracerebral contamination. (1)The single injection-indicator dilution method of Crone was adapted for use in the rat. A bolus containing a non-diffusible reference tracer and the test tracer was injected into the internal carotid artery. Simultaneously venous blood was sampled from the confluence of sinuses. It proved necessary to take special steps to avoid extracerebral contamination of the samples. Therefore the extracerebral branches of the carotid tree on both sides were ligated, and the venous sampling was done without suction. Results obtained in animals prepared surgically in different ways were compared. (2) 24Na+ was used to assess the degree of extracerebral contamination, the transcapillary loss of 24Na+ being a sensitive measure since the permeability of cerebral capillaries for sodium is about 4-5% that of muscle. (3) The transcapillary fractional loss, extraction E, was determined in this preparation with minimal extracerebral contamination. ENA+ was 2.9%, Eglucose was 12.5% at normal blood glucose levels whilst the net glucose extraction (calculated from arterio-venous difference) was about 10%. (4) It is concluded that extracerebral contamination is an important pitfall in indicator dilution studies on the brain. Use of 24Na+ as an indicator of extracerebral contamination is therefore recommendable.


Progress in understanding and treating Parkinson's disease. This review evaluates the long-term results of Levodopa therapy in Parkinson's disease upon quality of life, prolongation of survival and excess mortality. It also focuses on recent and new therapeutic approaches: Levodopa in combindation with a Dopa-decarboxylase inhibitor or MAO-B inhibitor, dopamine agonists and an active tripeptide: L-prolyl-L-leucylglycine amide (MIF-I). It ends by looking at new avenues of etiological research in Parkinson's disease which may indicate specific accelerated ageing of catecholaminergic (pigmented) neuronal systems.


Tinea cruris: diagnostic confusion due to isolation of Candida albicans alone. The diagnostic importance of the isolation of Candida albicans from a skin lesion is often uncertain. In a 68-year-old man from whose lesions only C. albicans was originally isolated Trichophyton rubrum and Epidermophyton floccosum were also isolated when the growth of the yeast was inhibited in a selective medium. The use of this selective medium, casamino acids erythritol albumin agar, ensures the proper interpretation of the significance of the presence of C. albicans in skin lesions.


Similarities of surface characteristics of neoplastic well-differentiated lymphocytes from solid tissues and from peripheral blood. Cells from spleens and lymph nodes infiltrated with neoplastic well-differentiated lymphocytes (WDLT) were investigated for the presence of receptors for unsensitized sheep red blood cells, complement (IgM EAC), and antigen-antibody complexes, as well as for the presence of surface immunoglobulins. In addition, the cells were examined by scanning electron microscopy (SEM). The surface markers and ultrastructure of WDLT cells were compared with those of cells obtained from peripheral blood of patients with chronic lymphocytic leukemia (CLL). Like the CLL populations, most WDLT samples showed a low percentage of unsensitized sheep red blood cell-binding cells and a concomitantly high percentage of IgM EAC-binding cells. Furthermore, a substantial number of WDLT cells in several instances showed a lower binding affinity for IgM EAC than normal lymphocytes, as demonstrated by the poor attachment of the IgM EAC reagent to the WDLT cells in suspension and to frozen tissue sections. Weak IgM EAC binding capacity was demonstrated in the CLL cells as well. WDLT and CLL cells bound the IgG EA complex only when certain sensitive techniques were applied (pelleting). A variable percentage of cells with surface immunoglobulins was found among WDLT and DLL populations. However, accurate counting of surface immunoglobulin-bearing neoplastic cells was very difficult in most cases due to the extremely faint surface fluorescence as compared to normal. SEM did not reveal specific changes in WDLT or CLL cells that would permit their identification as neoplastic lymphocytes. No major differences were found between WDLT and CLL cells and no correlation could be established between SEM appearances and surface markers. As previously observed with other cell populations, the conditions under which the cells were manipulated before fixation (in particular the environmental temperature) seemed to play a major role in the final SEM appearance of both WDLT and CLL cells. The results of our studies indicate that WDLT cells represent a population of B-cells whose surface properties are very similar to those of circulating CLL cells. The poor capacity of WDLT cells and CLL cells to bind IGM EAC or to stain with fluorescein-conjugated antihuman immunoglobulin suggests low density or poor affinity of the surface receptors. In this regard, these neoplastic cells are different from those of follicular lymphomas, also of B-cell origin.


Treatment of the mouse Lewis tumor by the association of radiotherapy and immunotherapy with Bacillus Calmette-Guérin. Three groups of 6 to 8-week-old male mice with transplanted C57BL/6 X DBA/2F1 tumor were irradiated by a single dose of 60Co localized on the tumor in association with i.p. injections of lyophilized Bacillus Calmette-Guérin. When the animals were sacrificed, tumor parameters (weight, surface area, and volume) were determined. Lung metastases were counted using a binocular magnifying glass. When Bacillus Calmette-Guérin injections were begun the day after the irradiation, tumor growth and the number of lung metastases were significantly decreased in comparison with the control group (p less than 0.01). However, there was no significant difference between the control group, which received radiotherapy alone, and the groups in which Bacillus Calmette-Guérin treatment was started after the 4th day postirradiation. This emphasizes the importance of timing in combined radioimmunological treatment.


The effects of cancer chemotherapeutic agents on normal hematopoietic precursor cells: a review. The effects of antineoplastic agents, singly or in combination, on normal hematopoietic precursor cells have been reviewed. Following a description of the assays used (e.g., spleen colony, in vitro colony, repopulating ability), the dose response and/or time response for each drug are presented by species and by assay as available. The schedule of drug administration, the time of the assay, and the proliferative state of the target population are the most important determinants. Alkylating agents, antitumor antibiotics, and 5-fluorouracil have exponential dose survival curves. "Phase-specific" agents such as antimetabolites, Vinca alkaloids, and ribonucleotide reductase inhibitors have plateaus in their dose survival curves, although the level of this plateau is different for different agents. Most drugs are more effective against rapidly proliferating cells although busulfan is less effective. Direct interspecies comparisons are possible with some of the clonogenic assays, which may allow prediction of the magnitude of human hematological toxicity for new agents or combinations.


Endotoxin enhancement of plasma cell tumor development in mice given injections of mineral oil. Bacterial endotoxins administered to BALB/c mice given i.p. mineral oil cause an increased incidence of plasma cell tumors, compared with mice given either oil or antigens alone, or oil plus antigen other than endotoxin. Endotoxin in ng doses was more effective than in mug doses.


Relationship between activity and amino sugar stereochemistry of daunorubicin and adriamycin derivatives. The effects of 4'-epi-daunorubicin, 4'-epi-adriamycin, and the corresponding beta anomers on the in vitro activity of Escherichia coli DNA polymerase I and RNA polymerase were determined and compared with the effects of the parent compounds. The observed effects parallel the cytotoxic activities, assayed by inhibition of mouse embryo fibroblast proliferation, and the inhibitory activities on DNA synthesis in cultured cells. The data indicate that the inverted configuration at position 1 of the amino sugar results in a markedly reduced biological activity. This conclusion is also substantiated by the data obtained with the beta anomer of adriamycin. A preliminary investigation on the binding properties of these derivatives suggests that the inverted configuration at C-1' produces a significant decrease in the binding to DNA. In contrast, epimerization at position 4' did not produce any significant change in activity. The relationship between biological and biochemical activity and DNA binding properties of the tested compounds are discussed with particularly reference to antitumor activity.


Immune response to an allogeneic progressively growing solid tumor. A C57BL/6 (B6) methylcholanthrene-induced solid tumor grows progressively in BALB/c mice. The BALB/c mice fail to develop in their spleens cytotoxic lymphocytes directed against the histocompatibility (H-) antigens of the tumor. A B6 skin graft is rejected normally by mice with the progressively growing B6 tumor and skin graft rejection has no influence on the growth of the tumor. Tissue culture-carried tumor cells work satisfactorily as target cells in a 51Cr release assay with effector lymphocytes immunized against B6 H-antigens, but the tumor cells will not work as an immunogen for H-antigens for an in vitro immunization. "Fresh" tumor cells removed from their in vivo environment and separated on a discontinuous bovine serum albumin gradient also work as target cells. The conclusion is drawn that the host cytotoxic lymphocyte response against the tumor, as defined by a short-term 51Cr release assay, suffers from both sensitization and effector stage defects that probably do not involve serum factors or suppressor cells.


Influence of castration followed by administration of LH-RH on the ultrastructure of rat pinealocytes. The ultrastructure of adult male rat pinealocytes was studied after orchidectomy and orchidectomy followed by LH-RH administration. Castration causes an increased development of the rough endoplasmic reticulum and the Golgi apparatus as well as an increase in the number of lipid droplets and lysosomes. The changes after orchidectomy followed by LH-RH administration were more marked than after castration alone. The ultrastructural features observed after castration can be interpreted as a morphological equivalent of increased protein synthesis. The observations reported appear to indicate mutual interaction between the pineal organ and the hypothalamo-hypophyseo-gonadal system. A working hypothesis is put forward implying the presence of a negative feedback mechanism between the pineal and the adenohypophysis.


The lateral photoreceptor of the barnacle, Balanus eburneus: quantitative morphology and fine structure. The lateral eye of the barnacle, Balanus eburneus, fixed in highly concentrated osmium is a lens-shaped body of approximately 250 mum in diameter and about 75 mum thick. It contains three photoreceptor cells which occupy about 42% of its volume. The photoreceptor cells are irregularly shaped and extend countless dendritic processes which bear rhabdomeres at their ends. Individual rhabdomeres come into contact with rhabdomeres originating from dendrites of the same or of one of the other visual cells. Thirteen per cent of the volume of the photoreceptor cells is taken up by the rhabdomeres. The membranes of the rhabdomeric microvilli contain globular subunits which suggest a 70 A spacing of rhodopsin molecules. There are two kinds of glial cells. One kind, type A glial cells, makes contact with the fibrous capsule of the photoreceptor. The other kind, type B glial cells, is associated with the photoreceptor cells and extends countless tiny cytoplasmic extensions which interdigitate with similar extensions of the receptor cells. There are approximately 95 type B glial cells and 130 type A glial cells in the receptor. The cytoplasm of the photoreceptor cells contains countless small Golgi fields, mitochondria, microtubules, multivesicular and multilamellar bodies. The extracellular space of the photoreceptor is less than 0.1% of its total volume.


The induction of filamentous growth in Escherichia coli by ruthenium and palladium complexes. Cis-chloroammine complexes of platinum have been shown to induce several biological effects. These include induction of filamentous growth in Escherichia coli as well as induction of phage production from lysogenic strains of Escherichia coli. Cis-chloroammine complexes of platinum have also been proven effective as a treatment for several animal tumors. In general, the effectiveness  of such compounds is determined by a combination of an internal polar nature coupled with an overall electrical neutrality. We have extended these studies to include other complexes of the group VIII elements. In the research reported here, complexes of ruthenium were shown to induce filamentous growth in E. coli. Cis-Ru(NH3)3Cl3 produced this effect at a concentration of 6 mug/ml, comparable to the required concentration of cis-Pt(NH3)2Cl2 for the same effect. The charged complex, K2RuCl5 - H2O, induced filamentous growth but a significantly higher concentration of this ruthenium complex was necessary. Several palladium complexes were also tested. They proved to be toxic at relatively high concentrations and had no effect  at lower concentrations. The proposed mechanism of action of the platinum complexes is the hydrolysis of two cis-chloride ligands followed by binding to DNA which reduces the synthesis of new DNA. The kinetic data on hydrolysis, the required cis-chloride ligands, electrical neutrality, and the selective action on various bacteria indicate that ruthenium and platinum interact by a similar mechanism.


Arterial and venous angiotensin II in normal subjects. Relation to plasma renin activity and plasma aldosterone concentration, and response to posture and volume changes. Plasma renin activity, arterial and venous angiotensin II (A II) concentrations, and plasma aldosterone concentration were measured in 16 normal subjects (mean age = 34 years) after 8 hours of recumbency, following 2 hours of ambulation, and again 30 minutes after administration of furosemide intravenously. Measurements were obtained during periods of sodium restriction and again during sodium surfeit. Both arterial and venous A II exhibited a 3-fold increase from low values of 8.8 +/- 2.5 and 8.6 +/- 2.5 pg/ml of plasma, respectively, during recumbency on high sodium intake to values of 23.9 +/- 4.1 and 26.5 +/- 6.2 pg/ml, respectively, following intravenous furosemide during sodium restriction. Corresponding values for aldosterone exhibited a 5-fold rise from 5.6 ng/100 ml to 32.0 ng/100 ml, whereas plasma renin activity (PRA) measured by an in vitro assay exhibited a 20-fold rise from 0.6 +/- 0.2 ng of angiotensin I (AI) generated per ml per hour to 13.1 ng/ml per hour. Despite the disparity in the magnitude of these increases, significant correlations were identified between all four of the measured parameters, indicating a major role of the renin-angiotensin system in regulating aldosterone output in response to volume and posture-related stimuli. Values of arterial and venous immunoreactive A II were closely correlated ( r = +0.72, P less than 0.005), but significant differences were demonstrated between low and high salt periods, suggesting that changes in metabolism of A II in the peripheral circulation may occur during sodium restriction.


The relevance of peripheral baroreceptors and chemoreceptors to regulation of cerebral blood flow in the cat. The contribution of neural vasomotor reflexes to the control of cerebral blood flow (CBF) was investigated in 30 cats lightly anesthetized with pentobarbital. CBF was measured both by kinetic analysis and by the intial slope technique of the washout curve of a bolus of 133Xe. Autoregulation (10 cats) and responsiveness to alteration in arterial PCO2 (PaCO2) (10 cats) and arterial PO2 (Pao2) (five cats) were assessed both before and after bilateral intracranial division of the 9th and 10th nerves. In an additional group (five cats), related changes in CBF to alteration of PaCO2 were recorded before and after unilateral section of the 7th and 8th nerves. Autoregulation was preserve after division of the 9th and 10th nerves and there was no significant change in the PaCO2 response curves. Section of the 7th and 8th cranial nerves did not produce conclusive results in the small number of cats studied. A conclusion that the facial nerves are not dominant in responses to hypercapnia seems justified, but a modulating role for these nerves is possible. These studies do not exclude a physiological role for these nerves in the autoregulation of CBF, but do indicate that the cerebral vascular bed apparently is capable of functioning normally after their division.


Role of heart and lung receptors with nonmedullated vagal afferents in circulatory control. Vagal afferents from the cardiopulmonary region exert a tonic inhibition on the vasomotor center. This is demonstrated by constriction of systemic resistance and splanchnic capacitance vessels and by increased output of renin when the vagi are cut or blocked. In dogs, removal or selective denervation of organs showed that receptors in the lungs, the atria, and the ventricles each are responsible for the vasomotor inhibition. That this inhibition is due to nonmedullated vagal afferents (C fibers) was demonstrated by selective cooling of the vagi, anodal block of medullated afferents, and selective electrical stimulation of medullated and nonmedullated fibers. In the open-chest cat the discharge frequency of individual C fibers is sparse and irregular (mean, 1.4 impulses/sec) but increases to 10 or more impulses/sec with moderate increases in cardiac filling pressure and exhibits cardiac rhythmicity or is continuous throughout the cardiac cycle. The inhibition of sympathetic vasomotor outflow effected through the cardiopulmonary receptors is inversely related to that exerted by the arterial baroreceptors. The former receptors have less influence on the muscle circulation than the latter, but have an equal or greater effect on the renal circulation. In summary, receptors in the heart and lungs with nonmedullated vagal afferents are an important component of the integrated neural control of the circulation.


Hemodynamic role of extracellular fluid in hypertension. Plasma volume is usually lower in patients with essential hypertension than in normal subjects. Normal or expanded plasma volume in some hypertensive patients may respect either a specific hypervolemic subset of the disease or the upper end of a continuum of volume values. Difficulties in defining these groups stem from the small numbers of subjects studied, from the need for a reliable reference index for volume measurements, and from the multiple factors which may affect intravascular volume. Differences in plasma volume cap influence choice of antihypertensive therapy; patients with expanded volume tend also to have slightly exchangeable sodium and greater extracellular fluid (ECF) volume and to respond well to diuretic therapy. There is also some evidence that low plasma renin activity is more frequent among hypervolemic patients. Essential hypertensives as a group have low plasma to interstitial fluid volume ratio (PV/IF), indicating that ECF distribution between the intravascular and interstitial compartments is shifted toward the latter. This is probably related to altered capillary filtration pressure due to increased venous resistance. Hypovolemic essential hypertensives have significantly lower (P less than 0.01) PV/IF ratio than hypervolemic, but whether this is related to differences in neural venous tone is only speculative. Hemodynamic studies revealed no difference in cardiac output between hypertensive patients with contracted blood volume and those with hypervolemia; total peripheral resistance was even higher in the latter, suggesting that "vasoconstriction" is not different between the two groups. It is widely believed that the relationship between ECF expansion and hypertension depends on the development of hypervolemia, increased cardiac output, and subsequent rise in total peripheral resistance reducing volume expansion and normalizing systemic flow while maintaining a high blood pressure. This sequence of events has been demonstrated in some human and experimental forms of hypertension but not in all. Metyrapone-induced hypertension in dogs could be sustained for up to 6 weeks by increased output with no evidence of "autoregulation" developing, and similar observations were reported in some anephric patients. Complementing these findings are observations of elevated cardiac output in some patients with long-standing essential hypertension or primary aldosteronism. It is therefore suggested that the spectrum of hemodynamic changes associated with volume disturbances in hypertension is too wide to be forced under one hypothesis alone and that neurogenic and other factors may play an important role in that complex relationship.


Measurement of circulating thyroid microsomal antibodies by the tanned red cell haemagglutination technique: its usefulness in the diagnosis of autoimmune thyroid diseases. Thyroid-microsomal antibodies were quantitated by a new technique utilizing tanned sheep red blood cells coated with human thyroid microsomal antigens. This haemagglutination assay (MCHA) correlated with the immunofluorescent antibody (FAB) but not with the thyroglobulin haemagglutination antibodies (TGHA) assay. Of forty-one patients with Hasmimoto's thyroiditis, thirty-nine (95%) were MCHA but only twenty-four (59%) TGHA positive. Titres were similar for the hypothyroid and euthyroid patients. Patients less than 20 years of age had either negative (50%) or low titre (less than 1:160) TGHA but 100% positive MCHA at titres greater than 1:1280. Of twenty-one patients with Graves' disease eighteen (86%) were MCHA and six (29%) TGHA positive. Of thirty-two patients without thyroid disease eleven (34%) were MCHA and/or TGHA positive. On the basis of family history and associated abnormalities, in eight of eleven, positive antibodies may have been due to subclinical Hashimoto's thyroiditis. Fourteen subjects of a control group (10%) were MCHA positive. Seven of ten examined had goitres. MCHA is a simple and quantitative test, useful in the diagnosis of autoimmune thyroid diseases.


Luteinizing hormone secretion in patients presenting with post-oral contraceptive amenorrhoea: evidence for a hypothalamic feedback abnormality. Twenty-four patients who presented with amenorrhoea after discontinuing oral contraceptives were studied. In fourteen patients underlying conditions were present which could account for the amenorrhoea. Two patients were pregnant, two had premature ovarian failure and four polycystic ovarian disease. Pituitary tumours were present in three patients, and in another three radiological abnormalities of the pituitary fossa suggested the presence of an intrasellar tumour. Basal serum luteinizing hormone levels reliably differentiated ovarian from hypothalamic or pituitary lesions. Varied patterns of serum LH response to clomiphene were present in the patients with post-oral contraceptive amenorrhoea or pituitary lesions. Most patients showed normal hormone response patterns, with a secondary LH peak, but in others a partial response only was seen. In the latter group, who responded normally to exogenous gonadotrophin releasing hormone, no secondary LH peak occurred despite normal oestradiol responses. This suggests an abnormality of the positive feedback system for oestrogens. Subsequent stimulation with clomiphene produced normal LH responses in these patients showing that the abnormality was reversible. These findings are consistent with the hypothesis that post-oral contraceptive amenorrhoea is the result of continuing suppression of normal hypothalamic pituitary feedback systems after the sex steroids have been stopped. The different patterns of hormone response to clomiphene suggest varying degrees of feedback abnormality, and in some patients the positive feedback mechanism only is impaired.


Modulation of pituitary responsiveness to exogenous LHRH by an oestrogenic and an anti-oestrogenic compound in the normal male. The effect of clomiphene (100 mg daily for 10 days) and ethinyl oestradiol (100 mug daily for 10 days) on the gonadotrophin response to synthetic LHRH has been investigated in two groups of five normal males. A third group of five men served as control group. LHRH, 25 mug, was injected intravenously on days 0, 4, 7 and 10 and the response of serum LH and FSH was monitored by radioimmunoassay. In contrast to the wide inter-individual variation of the response pattern, the intra-individual variation of the response to LHRH in the control group was small. Clomiphene induced a significant elevation of the baseline levels of LH and FSH after a few days of treatment; the pituitary responsiveness to LHRH, however, was significantly reduced. Oestrogen treatment resulted in a uniform suppression of both basal gonadotrophin levels and pituitary responsiveness. The decreased gonadotrophin response to LHRH during clomiphene treatment is thought to be caused by a relative and temporary pituitary depletion of the releasable gonadotrophin content. Although the suppression of LH and FSH response during oestrogen treatment may point to a direct inhibitory effect of oestrogen on pituitary gonadotrophin release, an indirect hypothalamic pathway, through suppression of endogenous LHRH, cannot be ruled out.


Immunofluorescence test for IgM rubella antibodies in whole serum after absorption with anti-gammaFc. A technique for detecting IgM rubella antibodies by immunofluorescence was developed. Total serum IgG was removed by prior immunoabsorption with anti-gammaFc. This prevents interference by rheumatoid factors in the serum. The test appeared sensitive and specific in sera of rubella convalescents and infants with congenital rubella.


Production of leucocyte inhibitory factor (LIF) and macrophage inhibitory factor (MIF) by PHA-stimulated lymphocytes. Supernatants from human mononuclear cells cultured with PHA inhibited the migration of both human polymorphonuclear leucocytes and guinea-pig peritoneal exudate cells, but not human mononuclear cells. Using ultrafiltration it was shown that these supernatants contained two inhibiting factors, the one with a molecular weight of 15,000-50,000 inhibited only guinea-pig peritoneal exudate cells (MIF), whereas the fraction containing molecules of a size between 50,000 and 75,000 specifically inhibited the migration of polymorphonuclear leucocytes (LIF). The polymorphonuclear leucocyte inhibiting activity was heat labile. It is suggested that the leucocyte migration inhibition test is dependent upon the production of a lymphokine (LIF) which acts specifically on polymorphonuclear leucocytes causing their inhibition of migration.


Antiarrhythmic effects of a lidocaine congener, tocainide, 2-amino-2',6'-propionoxylidide, in man. Eight patients with frequent premature ventricular contractions (PVCs) were given single oral doses of 2-amino-2', 6'-propionoxylidide hydrochloride (APX), a lidocaine congener, to investigate the drug's efficacy, toxicity, and pharmacokinetics. Of the 7 patients receiving more than 100 mg, 5 demonstrated at least 60% reduction in PVCs in the 5-hour period following one of the doses; the responsiveness to APX was similar to that observed when intravenous lidocaine was given to the same patients. There were side effects of transient dizziness, lightheadedness, and nausea in 3 patients 20 to 30 min after doses of 400 to 800 mg; slightly smaller doses in 2 of these patients retained efficacy without side effects. There were no effects on arterial blood pressure, heart rate, or the electrocardiogram of normally conducted beats. Laboratory values remained within normal limits, although the hemoglobin level and hematocrit showed a small but significant fall probably related to repeated blood sampling during the study. Drug plasma levels at 1 and 2 hr after administration and the area under the plasma concentration-time curve were proportional to dose, and the drug disappeared with a mean elimination half-life of 14.7 +/- 1.7 hr (mean +/- SD). Plasma levels resulting in suppression of PVCs ranged from 1 to 5 mug/ml. A mean of 39.7 +/- 12.4% of an orally administered dose was recovered unchanged in 48-hr urine collections. Comparison of the present results with those previously obtained after intravenous administration of APX indicates that oral bioavailability of the drug was essentially complete. APX appears to be a promising orally effective antiarrhythmic drug with suitable pharmacokinetic characteristics to warrant studies designed to establish dosage regimens for chronic therapy.


Antitussive activity of diphenhydramine in chronic cough. A double-blind crossover study was conducted to evaluate the antitussive effectiveness of diphenhydramine (DPH) in chornic coughs related to bronchitis, at doses of 25 and 50 mg every 4 hr for four doses. Both 25- and 50-mg doses caused a statistically and clinically significant reduction in frequency of coughs, compared to placebo. The most frequently reported side effect was drowsiness, principally at the 50-mg dose level. There was little or no apparent correlation between antitussive effectiveness and incidence of drowsiness, however, suggesting that the two effects were pharmacologically unrelated.


Quinine disposition during malaria and during induced fever. Quinine disposition was studied in 5 subjects before and during an experimentally induced infection with a chloroquine-resistant strain of Plasmodium falciparum and in 2 individuals before and during artificially induced fever. Plasma quinine levels were determined by both a benzene extraction method (QB), which measures principally unmetabolized quinine, and a metaphosphoric acid precipitation method (QMPA), which measures quinine and quinine metabolites. The ratio QB/QMPA in plasma was used to estimate the extent of metabolism of quinine. In all individuals plasma levels of quinien and QB/QMPA ratios were increased during malaria, suggesting impaired hepatic metabolism of quinine. The changes observed during malaria were not due to altered renal excretion of quinine. In 2 subjects in whom fever was artificially induced there were similar changes in quinine metabolism. These observations suggest that quinine dosage should be modified during the initial period of treatment, when symptoms and fever are greatest, in acute falciparum malaria.


Variations between individuals and populations in the acetylation of isoniazid and its significance for the treatment of pulmonary tuberculosis. Isoniazid is the most effective and widely used antituberculosis drug. The metabolism of isoniazid is noninducible and the primary metabolic route determining the rate at which isoniazid is eliminated from the body is acetylation. There are large diffrences between individuals in the rates at which isoniazid and certain other hydrazides and some sulfonamides are acetylated. The acetylation of isoniazid is inherited in a simple Mendelian fashion, individuals being either slow, heterozygous rapid, or homozygous rapid acetylators. Improved methods of discriminating between the different phenotypes have recently been described that are also more convenient to use. The proportion of slow acetylators among Japanese and Eskimos is about 10%; among Chinese about 20%; among Caucasians, Negroes, and South Indians about 60%. The results of a series of meticulously controlled clinical trials carried out in India, East Africa, Hong Kong, Singapore, Czechoslovakia, and Britain have shown the isoniazid acetylator status of tuberculosis patients treated with isoniazid-containing regimens is of no prognostic significance when treatment is given on daily basis. It may, however, be of significance when twice-weekly regimens are employed, especially in circumstances in which only a short period of initial daily chemotherapy is given, and where the companion drug employed during the twice-weekly continuation phase is relatively weak. The acetylator phenotype is, however, extremely important in once-weekly treatment, rapid acetylators always having fared considerably worse than slow acetylators on all the regimens evaluated so far. Experimental studies of the action of isoniazid against Mycobacterium tuberculosis in vitro and in vivo have enabled these findings to be understood.


Guanabenz effects on blood pressure and noninvasive parameters of cardiac performance in patients with hypertension. Guanabenz (2,6 dichlorobenzylidene amino guanidine acetate) is a new antihypertensive whose mechanism of action appears to involve both central alpha adrenergic stimulation leading to suppression of sympathetic nervous system activity from bulbar vasoconstriction centers as well as peripheral adrenergic neuron blockade. In this study, the drug was shown to produce a statistically and clinically significant decrease in blood pressure during a 4-wk placebo-controlled double-blind study. For three additional months continued efficacy and safety were shown under open conditions in 17 hypertensive patients. Mild sedation occurred, but there were no postural hypotension, tachycardia, evidence of sodium retention, gastrointestinal disturbances, or electrocardiographic (ECB) abnormalities. Noninvasive parameters of cardiac performance in 10 patients after single doses of guanabenz showed no significant changes. Although the numbers of patients were relatively small, the data suggest that this new drug may be a useful antihypertensive agent that warrants further investigation.