Nonviral helper factors for leukemia virus expression in mice.

Nonviral helper factors as well as murine leukemia viruses (MuLV) can increase the infectivity of Friend spleen focus-forming virus (SFFV) in mice which restrict its natural helper virus. Such factors have been isolated from cell-free extracts of human leukemic spleens, bacteria, and mycoplasmata. Unlike immunodepressants and target cell stimulators (which can also increase the pathogenicity of SFFV if applied before infection), nonviral helper factors are most efficient when they are injected with SFFV into helper virus-restrictive mice. In these respects the nonviral factors resemble MuLV. However, helper factors differ from MuLV by their inability to replicate in neonatal mice and by their greater stability to heat. Although the human leukemic helper (HuLV) extracts examined lack detectable mycoplasma or bacterial contamination, the helper assay in mice will be of limited value as a possible detection system for human leukemogenic viruses until a unique marker is found by which to distinguish HuLH activity from that of prokaryotic origin. This makes a systematic comparison among the nonviral helper factors all the more important.


Cell-mediated immunity in human acute leukemia.

The general cell-mediated immunological reactivity of patients with acute leukemia has been found to be intact, although it may be depressed by extensive disease or by chemotherapy. Patients with acute leukemia also have cellular immune reactivity against tumor associated antigens, as measured by skin tests for delayed hypersensitivity, lymphocyte stimulation, and 51Cr release cytotoxicity. Skin reactions to autologous and allogeneic crude membrane extracts of blast cells correlated with disease state, positive in many patients in remission and negative in most patients in relapse. Extracts of human lymphoid tissue culture cell lines derived from lymphomas or leukemia also gave positive reactions in patients with acute leukemia, and also in patients with lymphoma and nasopharyngeal carcinoma. The antigens detected in the skin tests with the lymphoid cell lines appear to be different from those associated with Epstein-Barr virus (EBV) and from those detected in the 51Cr release assay. Evidence is presented which suggests a complex variety of antigens on blast cells and on the cell lines. Although leukemia associated antigens were also detected by lymphocyte stimulation and by cytotoxicity assays, the results did not correlate with the skin tests nor with each other. The possible use of these assays for monitoring the chemotherapy and immunotherapy of acute leukemia patients is discussed.


L-1210 and human acute leukemia kinetics as related to therapy.

Comparative considerations are made between human acute leukemia (AL) and mouse transplantable L-1210 leukemia. The main kinetic parameters, such as the growth fraction (GF), growth rate, and cell cycle times, of both human and mouse diseases, are compared. The striking differences in cell kinetics and in response to treatment may be viewed as depending on different leukemogenesis mechanisms. Therefore, some improvement in human AL chemotherapy is considered possible both by researching a more rational employment of cytostatic drugs, and studying other animal models quite similar to the human disease, such as AKR leukemia.


Combination chemotherapy in Hodgkin's disease: relation to radiotherapy.

The results of modern radiotherapy for Hodgkin's disease are correlated to the stage of the disease. Five-year disease-free survival for pathologically staged patients can be estimated as follows: PS I & II A...90%, PS III A & IIIsA...60%, PS I & II B ...75%, PS III B & IIIsB ...40%. Combination chemotherapy of the MOPP (nitrogen mustard, vincristine, procarbazine, and prednisone) type can be successfully employed for patients who develop recurrent disease after total lymphoid irradiation. There have been several randomized clinical trials attempting to improve the curative potential of radiotherapy by combining modern chemotherapy with the irradiation as an adjuvant. The tolerance of patients to total lymphoid irradiation and chemotherapy will be presented. The Stanford trial, initiated 5 years ago (1968), has demonstrated prolonged disease free survival when both forms of treatment are used initially in certain stages of the disease. However, actual survival has not yet improved, probably because the chemotherapy is successful in inducing durable remissions for patients who recur after irradiation.


Virus infections in laboratories in Japan.

Virus infections in laboratories in Japan from 1967 to 1972 were studied. Major causative agents were influenza virus, Australian antigen, Rickettsia, and vaccinia virus. The cases reported were neither severe, nor lethal. It was suggested that these infections were caused either by aerosol or contact. Measures to prevent infection in laboratories are discussed.


Recent advances in the physiology of cardiovascular reflexes, with special reference to hypotension.

The basic response of the carotid and aortic baroreceptor areas to pressure stimuli has been known for some years (Heymans and Neil, 1958); stimulation of these areas (acute hypertension) leads mainly to bradycardia and hypotension; reduction in receptor activity (acute hypertension) results in an increase in heart rate and an increase in arterial pressure. In this review, comments have been made on the inadequacy of the "control theory" approach in describing the details of such mechanisms. Some recent evidence is presented of the differences between the effects from stimulation of the carotid and aortic areas, of the modifying influences of efferent sympathetic nerves to the two receptor areas, and of some evidence for central nervous "control" of the reflexes. The responses of the reflexes in the complex situation of haemorrhage, together with possible reflex actions from the ventricles, are discussed.


Urinary carcinoembryonic antigen (CEA)-like molecules and urothelial malignancy: a clinical appraisal.

A total of 190 patients being treated or followed up for urothelial carcinoma have been studied by the serial estimation of their urinary and plasma CEA levels. Only 46% of patients with a urothelial neoplasm present have a raised urinary CEA level. Infection or ileal conduit urine vitiate the result as they produce high CEA levels in the urine in the absence of any neoplastic disease. The accuracy of urinary CEA estimations is compared with that of cytology. Plasma CEA levels do not serve as a useful guide to the presence of extra-urinary tract tumour spread if taken as isolated readings. However, serial plasma CEA estimations may indicate that metastatic disease is present several months before its detection by the more usual clinical methods in a minority of patients.


Active immunotherapy as an adjunct to chemotherapy in the treatment of disseminated malignant melanoma: a pilot study.

In patients with disseminated malignant melanoma an optimal method of immunization with irradiated tumour cells was developed by reference to an in vitro assay for circulating specific serum inhibitors of cell mediated cytotoxicity. This immunization protocol consisted of the intradermal inoculation of 2 times 10(7) irradiated allogeneic melanoma cells admixed with 50 mug of percutaneous BCG. This method of immunization induced a significant but transient fall in the specific inhibitory effects of the sera on tumour directed cytotoxic activity of the patients' lymphocytes. In a pilot group of 30 patients with disseminated malignant melanoma being treated with chemotherapy (DTIC and vincristine) the immunotherapy was given midway between courses of the cytotoxic drugs. There was a correlation between the effects on circulating inhibitor and clinical outcome. The number of objective regressions occurring in this small pilot group was surprisingly high (17/30) and these clinical effects, although obtained in a series without concurrent controls, are presented for discussion. We suggest that the approach illustrated by this study, employing in vitro assays of tumour directed immune responses, may provide a suitable rational basis for the use of active immunotherapy as an adjunct to chemotherapy in the treatment of malignant disease.


Changes in host immunity following excision of a murine melanoma.

Changes in cell mediated and humoral immunity following the excision of a transplantable melanoma growing in the footpad of its syngeneic host, as measured by an in vitro cytotoxicity test, were assessed. Spleen cell cytotoxicity did not change significantly. Cells from the regional lymph nodes stimulated tumour growth before tumour excision. Three days following tumour excision this stimulatory effect was undetectable. Loss of serum factors capable of blocking the cytotoxicity of spleen cells occurred 24 h after tumour excision. Serum cytotoxicity increased after tumour excision to a maximum of the third day. Following tumour excision the rise in serum cytotoxicity and loss of regional lymph node tumour stimulation were concomitant with the loss of blocking activity.


The carcinogenicity of polycyclic hydrocarbon epoxides in newborn mice.

Benz(a)anthracene injected subcutaneously during the first 3 days of life caused a dose related increase in the incidence of liver and lung tumours in Swiss mice but over a similar dose range, the K region epoxide of benz(a)anthracene was less effective. Neonatally injected 7-methylbenz(a) was considerably more active than its K region epoxide in increasing the incidence of liver tumours in males. Both the parent compound and the epoxide slightly raised the incidence of lung tumours. Both chrysene and its K region epoxide increased liver tumour incidence but not lung tumour incidence. The K region epoxides of dibenz(a,h)-anthracene and 3-methylcholanthrene were without apparent effect on the incidence of liver, lung or other tumours despite indications from previously reported studies that the parent hydrocarbons are active at the same dose levels. The K region epoxide of phenanthrene had no effect on the incidence of any kind of neoplasm.


The spatial distribution of oesophageal carcinoma in the Transkei, South Africa.

Data on the incidence of cancer of the oesophagus in the Transkei for years 1965-69 are presented, age specific rates for the sexes discussed and the spatial relationship of well-defined regions of the high and low incidence demonstrated.


Photodynamic destruction of human bladder carcinoma.

Eleven human bladder carcinomata of different degrees of differentiation were implanted in mice immunosuppressed by thymectomy, anti-thymocyte serum and x-rays. Seven carcinomata grew well and one poorly and 3 produced mainly fibrous nodules in the mice. Normal human bladder tissues were grown from 4 other patients. The administration of a haematoporphyrin derivative (HpD), followed 24 h later by exposure to white light, caused marked destruction of tumours but little or none of normal bladder tissues. HpD or light alone caused no damage to tumours or normal tissues. It is suggested that photodynamic therapy may be applicable in the treatment of superficial transitional cell carcinoma of the bladder.


Two polypeptide chain constituents of the major protein of the cornified layer of newborn rat epidermis.

The insoluble component of stratum corneum of rat epidermis yields two major bands after extraction with 8 M urea-mercaptoethanol-dithiothreitol. The ratio of these two bands is about 1:1 in terms of protein stain intensity and S-[14C]carboxymethyl label. Both polypeptides were purified to homogeneity by DE-52-cellulose, sodium dodecyl sulfate hydroxylapatite C column chromatography, and preparative DodSO4-polyacrylamide gel electrophoresis. The heavier polypeptide contains 30% alpha helix and the lighter contains 27% alpha helix as determined by circular dichroism studies. Both are sensitive to Pronase and resistant to trypsin, collagenase, and elastase. The lighter chain is stable to pepsin but the heavier can be partially degraded to a smaller polypeptide with a molecular weight similar to that of light chain. Amino acid analysis shows that the light chain contains 12 more tyrosine residues than does the heavy chain, suggesting that the light chain is not generated from the heavy chain. However, the two chains may have a common peptide region. Antiserum prepared against the heavier polypeptide can be completely absorbed by purified lighter polypeptide and vice versa indicating that both chains have some common antigenic determinants. Antibody against either chain can cross-react with the stratum corneum and keratohyalin granules in the epidermis of newborn rat as indicated by fluorescent microscopic observation. Similarly, this antibody also cross-reacts with the cell surface or the contents of spinous and granular cells, and very weakly with basal cells, indicating that the two proteins may be present in the lower strata as well as the stratum corneum.


DNA-dependent RNA polymerases I and II from kidney. Effect of polyamines on the in vitro transcription of DNA and chromatin.

DNA-dependent RNA polymerases I and II were purified from pig kidney nuclei by chromatography on DEAE-Sephadex and phosphocellulose. When nonlimiting amounts of double-stranded DNA were used as the template, the in vitro transcription was markedly stimulated by spermidine and spermine. Maximal stimulation of RNA polymerase I occurred at 2-5 mM spermidine and 0.5-2 mM spermine, whereas optimal polyamine concentrations for RNA polymerase II were 5-10 and 1-5 mM for spermidine and spermine, respectively. DNA transcription by polymerase II was stimulated to a greater extent than that of polymerase I. Higher spermine (5-10 mM) concentrations were strong inhibitors of both polymerases under these conditions. The apparent Km of RNA polymerases I and II for UTP was unchanged at optimal polyamine concentration; under the same conditions the maximal reaction velocity was increased two- to three-fold and was essentially due to an increase in the rate of chain elongation. Thus, in a typical experiment the average chain length as determined by the UMP/uridine ratio increased from 570 to 1330 and the chain elongation rate increased from 0.64 to 1.44 nucleotides times sec-1 in the presence of spermine. When limiting quantities of native DNA were employed as the template, both RNA polymerases I and II were inhibited by 1-2 mM spermine. Kidney chromatin could be transcribed by homologous RNA polymerases with an efficiency ranging from 2 to 10% of that with native DNA. When chromatin was used in nonlimiting amounts instead of DNA, RNA polymerase II activity was again stimulated about two-fold at 2 mM spermine. Under these conditions, RNA polymerase I activity was inhibited by spermine. The inhibition of RNA synthesis in vitro at limiting quantities of templates (DNA or chromatin) could be overcome by preincubation of the enzyme with templates before polyamines were added. This inhibition thus appears to be due to a block in the initiation of RNA chains. Similar inhibition of transcription by RNA polymerase II was also observed with limiting quantities of chromatin as the template.


Mechanism of inhibition of RNA polymerase II and poly(adenylic acid) polymerase by the O-n-octyloxime of 3-formylrifamycin SV.

Factors affecting the inhibition of RNA polymerase II from rat liver by the O-n-octyloxime of 3-formylrifamycin SV (AF/013) were investigated. Using either native or denatured calf-thymus DNA as template, almost complete inhibition of RNA polymerase II was observed when AF/013 was added directly to the enzyme. Considerable resistance to AF/013 was observed when RNA polymerase II was preincubated with denatured DNA at either 0 or 37 degrees. However, under similar conditions, no resistance was observed when enzyme was preincubated with native DNA. Only when AF/013 was added to the ongoing reaction using native DNA did a resistance to AF/013 occur. The inhibition of RNA polymerase II by AF/013 was competitive with respect to all four nucleoside triphosphate substrates. The inhibition by AF/013 remaining after enzyme-DNA complex formation also appeared competitive with nucleoside triphosphate levels. The effect of exogenous protein (bovine serum albumin, BSA) on the inhibition of RNA polymerase II was also investigated. BSA reduced the extent of inhibition by AF/013, but did not alter the competitive nature of inhibition. Concurrently, the inhibition of highly purified nuclear poly(A) polymerase from rat liver, a template independent enzyme which incorporates AMP in a chain elongation reaction, was examined. As in the case of RNA polymerase, poly(A) polymerase was inhibited by AF/013 in a manner competitive with the nucleoside triphosphate substrate. The competitive nature of inhibition of RNA polymerase by AF/013 with respect to all four nucleoside triphosphate substrates, before and after enzyme-DNA complex formation, as well as the competitive nature of inhibition of poly(A) polymerase with respect to ATP tend to indicate that the major effect of AF/013 on RNA polymerase II is at the level of the substrate binding as opposed to a specific inhibition of initiation.


Porcine proparathyroid hormone. Identification, biosynthesis, and partial amino acid sequence.

Porcine parathyroid gland slices were incubated with 3H-labeled amino acids in order to label tissue proteins. After incubation a crude hormonal extract was prepared and analyzed by chromatography on carboxymethylcellulose. Among the three radioactive peaks which were detected in the eluate, two were identified as parathyroid hormone and proparathyroid hormone. Based on thin layer gel filtration in the presence of 6 M guanidine-HCl, the proparathyroid hormone had a molecular weight of 11,500 compared to about 9600 for parathyroid hormone. Radioisotope sequence analysis of the proparathyroid hormone revealed a partial sequence of: Lys1-Pro2-Ile3-Lys4-Lys5-Arg6-Ser7-Val8-Ser9--Ile11--Met14--Gly18--Ser22--Ser23---. Thus, from position 7 onward the relative position of each amino acid tested in this molecule corresponded exactly to that in the porcine parathyroid hormone sequence. The conservation of a similar, though not identical, basic hexapeptide grouping Lys-X-Y-Lys-Lys-Arg- at the amino terminal region of the prohormone in all species examined thus far (porcine, human, and bovine) suggests that this segment of the molecule may play an important role in the conversion of the prohormone to the hormone.


Replacement of acyl and alk-1-enyl groups in Clostridium butyricum phospholipids by exogenous fatty acids.

The effect of exogenous unsaturated fatty acids on the acyl and alk-1-enyl group composition of the phospholipids of Clostridium butyricum has been examined. Unsaturated fatty acids support the growth of this organism in the absence of biotin. When cells were grown at 37 degrees in media containing oleate or linoleate and a Casamino acid mixture containing traces of biotin, the exogenous fatty acids were found mainly in the alk-1-enyl chains of the plasmalogens with less pronounced incorporation into the acyl chains. However, at 25 degrees in this medium, both the acyl and alk-1-enyl chains contained substantial amounts of the 18:1 supplement plus the C19-cyclopropane chains derived from it. Ak-1-enyl chains in all the major phosphatide classes showed a uniformly high substitution by the oleate supplement in cells grown at 37 degrees. The oleate and C19-cyclopropane content of the acyl chains was more variable among the phosphatide classes. At 37 degrees, trans-9-octadecenoic acid (elaidic acid) also supported growth and was incorporated into both acyl and alk-1-enyl chains at a high level. When cells were grown on oleate at 37 degrees in media containing biotin-free Casamino acids, both the acyl and alk-1-enyl chains had a high level of 18:1 plus C19-cyclopropane chains. In the cells grown at 37 degrees with oleate substantial changes were seen in the phospholipid class composition. There was a large decrease in the ethanolamine plus N-methylethanolamine plasmalogens with a corresponding increase in the glycerol acetals of these plasmalogens. The glycerol phosphoglycerides were also significantly lower with the appearance of an unknown, relatively nonpolar phospholipid fraction.


Bromopyruvate inactivation of 2-keto-3-deoxy-6-phosphogalactonate aldolase of Pseudomonas saccharophila. Kinetics and stereochemistry.

The enzyme 2-keto-3-deoxy-6-phosphogalactonate aldolase of Pseudomonas saccharophila is inactivated by the substrate analog beta-bromopyruvate, which satisfies several criteria of being an active site directed reagent. The inactivation exhibits saturation kinetics, and both bromopyruvate and pyruvate (substrate) compete for free enzyme. Upon prolonged incubation, inactivation is virtually complete. The Kinact for bromopyruvate is 12 mM and the minimum inactivation half-time is 16 min with a k of 0.0433 min minus 1. Bromopyruvate is also a substrate for the enzyme in that 3(R,S)-[3-3H2]bromopyruvate is asymmetrically detritiated by the enzyme yielding 3(S)-[3-3H,H]bromopyruvate concomitant with inactivation. At various concentrations of bromopyruvate which affect the inactivation rate, the ratio of nanomoles of bromopyruvate turned over/unit of enzyme inactivated remains constant averaging 12:1, consistent with both inactivation and catalysis occurring at a single protein site, the catalytic site. The above value does not take into account a possible hydrogen isotope effect and is not thus an absolute value. The stereochemistry of bromopyruvate turnover catalyzed by this enzyme is the same as that for 2-keto-3-deoxy-6-phosphogluconate aldolase of P. putida. This fact provides the first evidence that the pyruvate-specific portions of the two active sites may have evolved from a common precursor.


Carbon-13 nuclear magnetic resonance studies of adenosylcobalamin and alkylcorrinoids, selectively enriched with carbon-13.

The carbon-13 nuclear magnetic resonance spectra of a series of alkylcorrinoids, selectively enriched with 13C in the alkyl ligand, were recorded at 25.2 MHz and 25 degrees. The nature of the axial ligands markedly affects the chemical shift of the labeled alkyl moiety (trans effect) as well as the 13C resonances of selected carbon atoms of the corrin ring (cis effect). Although a number of factors appear to influence the trans effect on the chemical shift of the alkyl ligand (important among them being electric field effects), the cis effect appears to be dominated by changes in charge density (at the methine bridge carbon atoms, C-5, C-10, C-15) and by steric effects (at the methyl groups at C-1, C-5, and C-15) accompanying axial ligation. Spin-latice relaxation times of several organocorrinoids, selectively labeled with 13C in the ligands attached to cobalt, were also measured. The T1 values of the methylene carbons of [5'-13C]adenosylcobalamin and [2-13C]carboxymethylcobalamin are very similar to that of the methine bridge carbon atom C-10 of the corrin ring, indicating that rotation about the carbon-cobalt bond of these two corrinoids is severely restricted. On the other hand, internal rotation about the carbon-cobalt bond of methylcobalamin is rapid.


Partial purification of a morphological transforming factor from pig brain.

A protein that changes one type of embryonic rat brain cell in culture from a primitive morphology to one resembling mature glial cell has been purified 400-fold from pig brain. The procedure includes differential centrifugation, ethanol treatment, trypsin digestion and column chromatography with Sephadex G-200 and Sepharose 4B. Although not completely homogeneous, the protein is biologically active at a concentration of 1-10(-8) M. It has a molecular weight of 350 000 and is heat labile. It is inactivated by the extremes of pH and by 8 M urea. The isoionic point is lower than neutrality. The activity is resistant to DNAase, RNAase, periodate and trypsin, but is susceptible to pronase digestion.


Physical properties and subunits of DNA dubia erythrocruorin.

The erythrocruorin of the freshwater leech Dina dubia possessed an S20,w of 61 S and exhibited a slightly sigmoid oxygenation curve with n congruent to 1.6 and P50 = 2.4 mm at pH 7.4. A minimum mol. wt of 23 000 +/- 2100 per heme group was determined from the iron and heme contents, 0.22 +/- 0.02 and 2.92 +/- 0.35 weight %. The subunit composition of this erythrocruorin was investigated using polyacrylamide gel electrophoresis and gel filtration in sodium dodecyl sulfate at neutral pH and gel filtration at pH 9. Sodium dodecyl sulfate electrophoresis of Dina erythrocruorin revealed the presence of five subunits (1-5) with mol. wts of about 13 000, 21 000, 23 000, 25 000 and 31 000, respectively. When the erythrocruorin was reduced with mercaptoethanol prior to dodecyl sulfate electrophoresis, three subunits (I-III) were observed, two possessing molecular weights in the range 12 000-14 000 (I and II) and one possessing a molecular weight of about 28 000. One of the subunits I, II, was provided by the dissociation of the 31 000 subunit. Subunit III (28 000) consisted of subunits 2, 3, and 4. It is likely that not all of the polypeptide chains constituting Dina erythrocruorin are associated each with a heme group.


Structural deterioration of tendon collagen in genetic muscular dystrophy.

The structure of gastrocnemius tendons from chickens with genetically induced muscular dystrophy has been studied by low-angle X-ray diffraction. Compared with normal samples there is poor alignment of collagen within the tendons. This difference is quite pronounced at eight weeks when the affected birds are still in comparatively good physical condition. Similar changes have been reported for birds with nutritionally induced muscular dystrophy (Bartlett, M. W., Egelstaff, P. A., Holden, T. M., Stinson, R. H. and Sweeny, P. R. (1973) Biochim. Biophys. Acta 328, 213-220).


Kinetic analysis of the polymerization process of actin.

The polymerization process of actin was examined by measuring the amount of flow birefringence and by analyzing release of labeled inorganic phosphate from the bound [gamma-32P]ATP upon polymerization of G-actin to F-actin. Comparison of the above experimental results with the electron microscopic data of Kawamura and Maruyama (J. Biochem., 67, 437-457, 1970) suggested that growth and redistribution steps occurred simultaneously during polymerization. Attempt was made to simulate the polymerization process of actin by calculating the kinetic equations numerically. The results of simulation suggested that it was necessary to take into consideration the association and dissociation between F-actin particles as well as the association and dissociation between F-actin and G-actin.


Molecular weight of Eudistylia vancouveri chlorocruorin and its subunits.

The chlorocruorin of the marine polychaete Eudistylia vancouveri has a molecular weight of 3.1-10(6) and a sedimentation coefficient (S020, w) of about 57 S at pH 8.0 in the presence of 0.01 M Mg2+. The quaternary structure of this pigment is unaffected by pH between 6.0 and 11.5 in the presence of 0.01 M Mg2+ whereas in 0l01 M EDTA, the pigment begins to dissociate above pH 9.0 into smaller submultiples. The chlorocruorin can be converted into subunits with molecular weights of about 14 000-15 000 and 30 000 as determined by sodium dodecyl sulfate-gel electrophoresis and 14 000-15 000 as measured by gel chromatography of the carboxy-methylated derivative in 8 M urea, 0.1 M 2-mercaptoethanol, or by sedimentation equilibrium in 6 M guanidine-HCl and 0.1 M 2-mercaptoethanol. The pigment contains 0.212 +/- 0.008% iron corresponding to 1 g atom iron per 26 300 g chlorocruorin. The amino acid composition of this pigment is reported. The subunit structure of Eudistylia chlorocruorin and the polymeric annelid hemoglobins are similar in many respects.


Snake venom toxins. The primary structure of protein S4C11. A neurotoxin homologue from the venom of forest cobra (Naja melanoleuca).

Six minor protein constituents (S4C10-S4C15) have been isolated from the venom of Naja melanoleuca. The complete amino acid sequence of S4C11 has been established and indicates that it is a homologue of the neurotoxins which are found in elapid venoms. The other proteins appear from the amino acid compositions to be homologues of the cyto- or cardiotoxins found in cobra venoms. Protein S4C11 has a low toxicity, failing to kill mice at an intravenous dose of 20 mug/g body weight. The sequence of the first 25 residues out of the total of 65, was determined using the automatic sequenator. The remainder of the sequence was derived with the aid of tryptic and chymotryptic peptides. The sequence showed the unusual feature of having 65 amino acid residues including 10 half-cystine residues.


Hemoglobin Fort Gordon or alpha2beta2145 Tyr replaced by Asp, a new high-oxygen-affinity hemoglobin variant.

Hemoglobin Fort Gordon, alpha2beta2145 Tyr replaced by Asp (HC2), has been observed in a 20-year-old black male with compensatory erythrocytosis. The variant was readily identified by electrophoresis and chromatography, and comprised about 30% of the red cell hemoglobin. The substitution was identified through analyses of tryptic peptides of various digests of the isolated beta chain. The oxygen affinity of whole blood was increased; two components were observed one of which had a greatly increased affinity for oxygen and a markedly reduced subunit cooperativity. It appears that the Tyr replaced by Asp substitution resembles the Tyr replaced by His substitution in hemoglobin Bethesda (Bunn, H. F. et al. (1972) J. Clin. Invest. 51, 2299-2309; Olson, J. S. and Gibson, G. H. (1972) J Biol. Chem. 247, 3662-3670; Adamson et al. (1972) J. Clin. Invest. 51, 2883-2888) in that both inhibit the quarternary change of the oxy to the deoxy conformation, resulting in greatly altered functional properties. Studies of a few members of the family were negative.


Hemoglobin Wood beta97(FG4) His replaced by Leu. A new high-oxygen-affinity hemoglobin associated with familial erythrocytosis.

The characterization of hemoglobin Wood (beta97(FG4) His replaced by Leu), a high oxygen affinity hemoglobin with reduced Hill constant is described. The amino acid substitution occurs at the alpha1beta2 interface, in the same position as in hemoglobin Malmö (beta97(FG4) His replaced by Gln) and in an homologous position when compared with hemoglobins Chesapeake (alpha92(FG4) Arg replaced by Leu) and J. Capetown (alpha92(fg4) arg replaced by Gln).


Haemoglobin Titusville: alpha94 Asp replaced by Asn. A new haemoglobin with a lowered affinity for oxygen.

A new haemoglobin variant with a decreased oxygen affinity is described, in which the substitution, alpha 94 (G1) Asp replaced by Asn, affects the alpha1beta2 contact alpha1G1-beta2G4. The relevance of this variant to our understanding of the importance of the hydrogen bond between alpha1G1 and beta2G4 in Perutz's model of oxyhaemoglobin A is discussed.


Studies on phytohemagglutinins. XXV. Isolation and characterization of hemagglutinins of the spindle tree seeds (Evonymus europaea L.).

A mixture of isophytohemagglutinins has been isolated from the fleshy arils of the spindle tree seeds (Evonymus europaea L.) by fractional precipitation of the saline extract of the arils by (NH4)2SO4 at a 0.40% saturation. Successive preparative disc electrophoresis on polyacrylamide gel affords separation of one slower moving component, phytohemagglutinin I, from the mixture of other isophytohemagglutinins that have a very similar electrophoretic mobility. Phytohemagglutinin I has a sedimentation coefficient Sw,20 of 7.1 S and an approximate mol. wt of 127 000. Amino acid analysis shows a high amount of aspartic acid, alanine and glycine but also significant amounts of serine, threonine, cysteine and methionine. Aspartic acid is the only N-terminal amino acid found by the dansylation technique. Phytohemagglutinin I contains glucosamine and 4.7% neutral sugar. Its approximate pI in citrate/phosphate buffer is 4.4-4.5. The metal content amounts to 0.250% Ca, 0.019% Mg, 0.034% Zn and 0.026% Cu. Mn is not present. Ultracentrifugation analysis reveals homogeneity in the sedimentation behavior of the mixture of isophytohemagglutinin, an Sw,20 of 7.1 S and an approximate mol. wt of 119 000. The mixture has an amino acid composition closely resembling that of phytohemagglutinin I and an identical pI but contains only 1.9% neutral sugar. Two N-terminal amino acids were shown to be present, aspartic acid and tyrosine. With the exception of Cu which is absent, the metal content is almost the same as that of phytohemagglutinin I. Both phytohemagglutinin I and the mixture are devoid of anti-A1 activity and show detectable anti-H, anti-B and anti-A2 erythroagglutinating activity in approximate limit concentrations of 2.5, 5 and 10 mug/ml, respectively. This activity is not influenced by the presence of EDTA, Ca2+ or Mg2+, but is stimulated by Zn2+. Mn2+ and Co2+ have an inhibitory effect. None of the simple sugars tested inhibited the hemagglutination reactions.


Interaction of rabbit hemoplexin with copro- and uroporphyrins.

Rabbit hemopexin forms equimolar complexes in vitro with the I and III isomers of both coproporphyrin and uroporphyrin. The apparent dissociation constants (Kd) of these complexes are estimated to be 4-10(-7) M for coproporphyrin-hemopexin and 10(-6) M for uroporphyrin-hemopexin by equilibrium dialysis and quenching of protein fluorescence. Results of competitive binding experiments suggest that all four porphyrins bind at the heme-binding site of hemopexin, and that the relative affinity of rabbit hemopexin for these porphyrins is: deuteroheme greater than coproporphyrin I or III greater than uroporphyrin I or III. These findings provide further evidence that hemopexin may function as a transport protein for circulating coproporphyrins as well as for heme.


Enzymic and immunochemical properties of lysozyme. IX. Conformation and immunochemistry of derivatives succinylated at certain lysine residues.

Succinylation of lysozyme in the presence of 7 molar excess of [1,4-14C2]-succinic anhydride gave a reaction product which showed at least six components by disc electrophoresis. Chromatography on CM-cellulose enabled the isolation of six homogeneous derivatives. The derivatives were succinylated at the following locations: derivative I, lysines-1 (alpha- and epsilon-NH2), -13, -97 and -116 and the OH group at position 43 (or 36 or 40); derivative II, lysines-1 (alpha- and epsilon-NH2), -13, -96, -116; derivative III, lysines-1 (alpha-and epsilon-NH2), -13, -97, -116; derivative IV, lysines-1 (alpha-NH2), -33, -96 and -116; derivative V, lysines-1 (alpha-NH2), -33 and -96; derivative VI, lysines-33 and -116. Conformational changes were detectable in derivative I by ORD and CD measurements and by accessibility of the disulfide bonds to reduction. On the other hand, the other five succinyl derivatives showed no conformational changes by ORD and CD measurements. However, their disulfide bonds were slightly more accessible to reduction than lysozyme, with the increase being somewhat higher in derivatives I, II and III. Enzymic activity measurements showed that only derivative VI possessed some (10%) enzymic activity. Immunochemical studies with antisera to lysozyme showed that the reactivity of each of the derivatives was lower than the homologous reaction. Correlation of the extent of decrease in immunochemical reaction with the locations of modification and with the results of conformational analysis, led to the conclusion that lysines 33, 96 and 116 are part of antigenic reactive regions in lysozyme. The modification results are also discussed in relation to the three-dimensional structure of lysozyme in solution.


The essential histidine residues of bovine plasma amine oxidase.

Ethoxyformic acid anhydride and photooxidation have been used to study the function of histidine residues in bovine plasma amine oxidase. Ethoxyformic acid anhydride at pH 6.1 reacted with nearly all of the histidine residues in the enzyme in 15 min but complete enzyme inactivation occurred in several minutes. The concentration of the reagent which caused 50% inhibition was 2.2-10(-5) M under the conditions of the experiment. The diamine oxidases, Aspergillus niger and pea seedling amine oxidases were also inhibited by ethoxyformic acid anhydride. The concentrations of reagent required for 50% inhibition were 6.6-10(-5) and 3.3-10(-4) M, respectively, for the two enzymes. NH2OH could not be used to regenerate the reacted histidine residues since NH2OH itself inhibited the enzyme. Photooxidation in the presence of 0.001% Rose Bengal at pH 7.0 also inactivated bovine plasma amine oxidase. Histidine was the only amino acid destroyed by photooxidation. About six histidine residues were destroyed but in the presence of the substrate kynuramine, two less histidine residues were destroyed. Since lysine which is neither a substrate nor inhibitor of the enzyme did not protect the enzyme from photooxidation, it was concluded that two histidine residues, one in each sub-unit of the enzyme are essential for activity.


An evolutionary comparison of plant histones.

Histones were extracted from chromatin of the following: a moss (Polytrichum juniperinum); the primitive vascular plants Psilotum nudum and Equisetum arvense; a fern (Polypodium vulgare); the gymnosperms fir (Abies concolor), yew (Taxus canadensis) and Gingko biloba; the dicotyledonous angiosperms tobacco (Nicotiana tabacum) and maple (Acer saccharinum); and the monocotyledonous angiosperms corn (Zea mays) and lily (Lilium longiflorum). The histones were subjected to polyacrylamide gel electrophoresis and compared to standard histones of pea (Pisum sativum) and cow (Bos taurus). All species have histones of the exact electrophoretic mobility of histones F2a1 and F3 of cow and pea. All species have histones of low electrophoretic mobility assumed to be F1 histones. None of the plant histones displayed electrophoretic mobility between F3 and F2a1 while animal histone fractions F2b and F2a2 do migrate to this position. No animal histone fraction was found to migrate between F3 and F1 while a major plant fraction, designated "F2b-like" was found to migrate to this position in all plant species studied except for the moss and Psilotum. A band of similar mobility was strikingly absent from the histones of these two species.


Free and membrane-bound chloroplast polyribosomes Chlamydomonas reinhardtii.

Over half of the chloroplast ribosomes isolated from growing cultures of Chlamydomonas reinhardtii are bound to chloroplast thylakoid membranes if completion of nascent polypeptide chains is prevented by chloramphenicol. The free chloroplast ribosomes are recovered in homogenate supernatants, and presumably originate from the chloroplast stroma. Only about 10% of these free chloroplast ribosomes are polyribosomes, even under conditions when 70% of free cytoplasm ribosomes are recovered as polyribosomes. The nonionic detergent Nonidet P-40 liberates atypical polyribosomes (Type I), from membranes, which require both ribonuclease and proteases for complete conversion to monomeric ribosomes. Thus Type I particles are held together by mRNA but are also held together by peptide bonds. These Type I polyribosomes probably are not bound to intact membrane, but might be bound to some protein-containing sub-membrane particle. The Type I polyribosomes are dissociated to ribosomal subunits by puromycin and high salt, and contained 0.2 to 1 nascent chain per ribosome. If membranes are treated with Nonidet and proteases at the same time, polyribosomes which are digested to monomeric ribosomes by ribonuclease alone (Type II) are obtained. Type II polyribosomes are smaller than Type I, and probably represent the true size distribution of polyribosomes on the membranes. At least 50% of the membrane-bound ribosomes are polyribosomes, since that much membrane bound chloroplast RNA is recovered as Type I or Type II polyribosomes.


The effect of a mammalian repair endonuclease on x-irradiated DNA.

DNA was extracted from rat liver of non-irradiated animals, and was irradiated in vitro, and from animals which received whole body doses of X-radiation. Sedimentation on neutral and alkaline sucrose gradients as well as measurements of 32P release after sequential treatment with endonuclease and alkaline phosphatase and determination of triphosphate incorporation after the sequential treatment with endonuclease, alkaline phosphatase and DNA polymerase indicated that DNA irradiated in vivo and in vitro were effective substrates for the mammalian repair endonuclease. The experiments suggest that in addition to strand breaks, X-radiation causes base damage and they have provided a plausible explanation for the formation of double strand breaks in DNA irradiated in vivo.


Abnormal synthesis of mitochondrial DNA in the presence of N-methyl-N-nitro-N-nitrosoguanidine in vitro.

1. The in vitro effect of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) on mtDNA synthesis was studied using isolated newborn rat liver mitochondria. 2. From the kinetics of the incorporation of [3H]thymidine into the acid-insoluble material, MNNG neither stimulated nor inhibited the DNA synthesizing activity of mitochondria. The activity observed in the presence of MNNG was inhibited by N-ethylmaleimide and actinomycin D. 3. By the band velocity sedimentation in CsCl/ethidium bromide, the properties of the nascent mtDNA formed in the presence of MNNG were analyzed. The nascent DNA-containing molecule was not found in the closed-circle fraction, and essentially detected in the open-circle fraction. This change of the template was blocked by N-ethylmaleimide but not by actinomycin D, suggesting a conversion of the closed-circular template to the open-circular one by single-strand cleavage(s). From the band sedimentation in alkaline CsCl, the number of nascent higher molecular DNAs was increased but the molecules were all of relatively lower molecular weight. On the other hand, the formation of nascent fragments was inhibited. 4. The alkaline CsCl equilibrium centrifugation analysis revealed that the nascent DNA synthesized in the presence of MNNG consisted of both light and heavy components. 5. Present results suggest that MNNG exerts its effect on the mtDNA synthesis by modifying the intrinsic mechanism of discontinuous synthesis, since the conversion of the template DNA molecule from the closed- to open-circular form and the continuous polymerization of the nascent higher-molecular DNA on such a relaxed template were characteristic events in vivo.


The molecular size and conformation of the chloroplast DNA from higher plants.

Covalently closed circular choloroplast DNA (ctDNA) molecules have been isolated from pea, bean, spinach, lettuce, corn and oat plants by ethidium bromide/cesium choloride density-gradient entrifugation. As much as 30-40% of the total ctDNA could be isolated as closed circular DNA molecules and up to 80% of the total ctDNA was found in the form of circular molecules. The size of pea, spinach, lettuce, corn and oat ctDNA relative to an internal standard (phiX174 replicative form II monomer DNA) was determined by electron microscopy. The ctDNAs showed significant differences in their sizes, and their molecular weights ranged from 85.4 - 10(6) for corn ctDNA to 96.7 - 10(6) for lettuce ctDNA. Each of these ctDNAs contained 3-4% of the circular molecules as circular dimers and 1-2% of the circular molecules as catenated dimes. The molecular complexity of these ctDNAs was studied by renaturation kinetics using T4 DNA as a standard. The molecular weights of the unique sequences of the ctDNAs ranged from 83.7 - 10(6) for oat ctDNA to 93.1 - 10(6) for lettuce ctDNA, which are in excellent agreement with the sizes of the circular ctDNA molecules...


Hormonal control of transcription in higher plants.

1. Nucleolar RNA polymerase Ib obtained from auxin-treated lentil roots exhibits a higher transcriptional activity than the enzyme obtained from control roots. This difference is due to a change in the enzyme properties after auxin treatment. It is suggested that the hormonal effect is mediated by a factor that changes the molecular properties of nucleolar RNA polymerase. 2. Four fractions, alpha, beta, gamma and delta, that stimulate the activity of RNA polymerase Ib, have been extracted from lentil roots. Two of them, gamma and delta have been studied. Factor delta can stimulate nucleolar polymerase Ib and the nucleoplasmic enzyme II equally well, while factor gamma is specific for polymerase Ib. 3. The curve of UMP incorporation in vitro, with and without factors gamma or delta suggests that they are initiation factors. This conclusion is reinforced by the analysis of simultaneous incorporation of [gamma-32P]ATP and [3H]UMP in the RNAs synthesized in vitro. 4. Although the level of factor delta is independent of auxin treatment, that of factor gamma is doubled in auxin-treated roots. These results suggest that factor gamma is an auxin-induced protein that modulates the specific activity of the nucleolar RNA polymerase. 5. A general model of the mode of action of auxins at the molecular level is proposed. It integrates into a unified scheme the above results as well as those obtained by other workers.


13C magnetic resonance investigation of mercury (II) binding to nucleosides and thiolated nucleosides in dimethyl sulfoxide.

Natural abundance, proton-decoupled 13C magnetic resonance spectroscopy is shown to be a useful technique for identifying the mercury (II) binding sites on nucleosides and especially thiolated nucleosides. Measurements made on dimethyl sulfoxide-d6 solutions, 0.5 M in nucleoside and 0.15 M in mercury, reveal that both CH3 HgCl and HgCl2 bind principally to the sulfur atoms of s6 Guo and s8 Guo. The 13C NMR spectra of the unthiolated nucleosides in the presence of excess (4:1) mercury reveal that HgCl2 binds to N-3 of cytidine, to more than one site on adenosine and guanosine, but not strongly to uridine. Excess HgCl2 shifts the thiocarbonyl carbon atoms in s6 Guo and s8 Guo approx. 16 ppm upfield compared to the free nucleosides, and there is evidence for additional coordination to N-7 of s6 Guo. Binding to the ribose hydroxyl groups is clearly ruled out. At least in these instances, 13C NMR proves to be useful for assigning the mercury (II) binding sites, complementing the results of proton magnetic resonance studies. Proton NMR data for the binding of CH3 HgCl and HgCl2 to s6 Guo and s8 Guo are also presented.


A procedure for the detection of pollution by fish movements.

An "alarm" system using fish movements has recently been developed to provide rapid information about accidental spills of pollutants. The data are the realization of a non-stationary stochastic process which takes on discrete values. By the use of the Camp-Paulson approximation, the process is transformed to a strictly stationary one for which calculations are tractable. A procedure is then presented to detect the presence of pollutants. Tests with real data indicate that it detects them sooner than the method used previously.


Transition densities for neutral multi-allele diffusion models.

Kimura [1955, 1956] partially solved the problem of finding the transition density function which describes the behavior of a diffusion model for evolution at one genetic locus with many neutral alleles. We complete the solution using a system of polynomials biorthogonal to polynomials suggested by Appell [1881] as generalizations of Jacobi polynomials.


Variances of selection differentials in normal samples.

Standardized variances of selection differentials are defined in general and then considered in detail for the case of directional selection on the basis of normally distributed test scores. Symmetry properties, bounds and asymptotic values for the variances are derived. The results are used to investigate the variance of response to selection.


The effect of incomplete follow-up.

By generating sampling distributions for each of three commonly used estimators for qz, the conditional probability of death in an interval, it is shown that the inclusion of subjects due for withdrawal introduces bias in all three, especially the "actuarial estimator" (Berkson and Gage [1950]). The bias is shown to depend on the proportion due for withdrawal and on the level of qz. Some practical suggestions are made for dealing with the bias.


Confidence intervals for the conditional probability of misallocation in discriminant analysis.

In this study we are concerned with the construction of confidence intervals for the conditional probability of misallocation associated with Anderson's classification statistics, W. The available methods of computing confidence intervals for the conditional probability are not satisfactory in practice, mainly because the intervals obtained are fairly inaccurate. A new method is presented which enables intervals with almost the desired level of confidence to be easily computed from the initial samples on which W is based.


Theory of inbreeding and covariances between relatives under full-sib mating in diploids.

A generation matrix theory of full-sib mating is developed in which 13 mating "classes" are distinguished according to identity of genes in individuals mated and identity of genotypes as belonging to homozygous, parental, or offspring sets. The 13 times 13 matrix reveals some properties of the full-sib mating system not shown by previous work. The eigenvalues and a set of eigenvectors for the generation matrix, and the general solution for the frequencies of mating classes among descendants of an original mating of genotypes ab times cd, are given. The genotypic array of descendants in an arbitrary generation is also given. A new formula is derived for the coefficient of inbreeding in generation n + m in terms of coefficients of inbreeding in earlier generations. An algorithm is presented for calculating the probability of a given situation of identity of alleles carried by two individuals given only the indices of their own respective generations and the generation of their most recent common ancestor. The application of such probabilities to obtaining covariances between relatives in a full-sib mating system, under the assumptions of independence and non-interaction among loci, is illustrated. All results are shown to agree with previous work in special cases. All possible full sib, generation n - 1 parent-generation n + m offspring, and generation n uncle-generation n + m nephew covariances for 1 less than n + m less than or equal to 8 are obtained using the given algorithm.


The coverage of a random sample from a biological community.

A taxonomic group will frequently have a large number of species with small abundances. When a sample is drawn at random from this group, one is therefore faced with the problem that a large proportion of the species will not be discovered. A general definition of quantitative measures of "sample coverage" is proposed, and the problem of statistical inference is considered for two special cases, (1) the actual total relative abundance of those species that are represented in the sample, and (2) their relative contribution to the information index of diversity. The analysis is based on a extended version of the negative binomial species frequency model. The results are tabulated.


A review and critique of some models used in competing risk analysis.

We have introduced a notation which allows one to define competing risk models easily and to examine underlying assumptions. We have treated the actuarial model for competing risk in detail, comparing it with other models and giving useful variance formulae both for the case when times of death are available and for the case when they are not. The generality of these methods is illustrated by an example treating two dependent competing risks.


Index selection with proportionality constraints.

Tallis [1962] formulated and solved the problem of constructing a selection index whose use will produce changes in the means of genotypic values of various traits that are proportional to ultimately desired changes. Several meaningful variations of this problem are described, and their solutions are presented in a common framework.


Quasi-independent triangular contingency tables.

The model of quasi-independence in a triangular contingency table is parametrised in a way which is both easy to interpret probabilistically, and easy to analyse.


On the construction of the population projection matrix for a population grouped in unequal stages.

It is shown how it is possible to construct a projection matrix for a population grouped in unequal stages directly from a known projection matrix for the population grouped in equal age intervals.


A multiple range procedure for correlated variances in a two-way classification.

A multiple range procedure is proposed for specifying homogeneous subsets of variances in a two-way classification. Appropriate critical values are specified and an example is provided for illustrative purposes.


A step-up procedure for selecting variables associated with survival.

Multivariate concomitant information on a subject's condition usually accompanies survival time data. Using a model in which each subject's lifetime is exponentially distributed, this paper suggests a method which utilizes a step-up procedure for choosing the most important variables associated with survival. Maximum likelihood (ML) estimates are utilized, and the likelihood ratio is employed as the criterion for adding significant concomitant variables. An example using multiple myeloma survival data and sixteen concomitant variables is discussed in which three variables are chosen to predict survival.


Effect of sex hormones on rabbit arterial subendothelial connective tissue.

The effect of sex hormones on subendothelial connective tissue in rabbit arteries was sought, and striking morphological alterations were noted after prolonged treatment. Depot preparations of testosterone cypionate, 100 mg/week, or estradiol cypionate, 200 mug/week, were injected intramuscularly into intact adult New Zealand male rabbits for 8-16 weeks. The iliac arteries were prepared for electron-microscopic examination from five rabbits in each treatment group and in four normal male rabbits. Thin sections were prepared from three randomly selected cross-sectional tissue blocks for each animal. A minimum of 7 mum of vessel circumference was studied in each block. Selection of an area for electron-microscopic examination was random, except that those sites in which the internal elastic lamina showed fragmentation were discarded. Electron micrographs at a single final magnification were evaluated 'blind' by several observers for qualitative difference in subendothelial constituents. In the control group, the subendothelium contained moderate numbers of elastin-associated microfibrils (MF); all the rabbits given testosterone consistently showed significantly fewer MF than the control. In contrast, rabbits given estradiol all had significantly greater numbers of MF. Statistical analysis of variance showed these difference to be significant at p less than 0.001.


Concerning the role of the amine pump of the adrenergic innervation of rabbit aorta in sustaining the neuron blockade produced by bethanindine and bretylium.

Adrenergic neuron blockade produced in rabbit aortic strips by bretylium and bethanidine has been studied. Differences were noted in the characteristics of approximately equal, strong neuron blockade produced by bretylium was readily reversed by washout of the drug from the tissue bath whereas blockade produced by bethanidine was slowly reversed. Desmethylimipramine prevented the onset of blockade from bretylium whereas it only delayed the onset of blockade due to bretylium but only slightly antagonized an established blockade due to bethanidine. It is suggested that the differences observed between these two neuron blocking agents are the result of their differential retention inside the neuron: bretylium is not firmly bound, leaks out of the neuron and goes through a process of recycling across the cell membrane, while bethanidine is more firmly bound inside the neuron than bretylium, only slowly leaks out of the neuron and is not recycled back across the cell membranes.


Evidence for non-mast cell histamine in the vascular wall.

Histamine has been found biochemically in the mammalian vascular wall. This study was undertaken in an effort to identify the presence of specific histamine-containing cells inwalls of large blood vessels such as femoral artery and vein, brachial artery and vein, mesenteric artery and vein, renal artery and vein, and aorta. Segments of vessels from rats,  cats, dogs, sheep, cows, humans and chickens were examined. Sections were prepared from fresh tissue by vibratome, from frozen tissue by cryostat and from freeze-dried and fixed tissues. Mast cells were visualized by staining with acidic toluidine blue and by reaction with orthophthaladehyde to develop histamine fluorescence. Although mast cells were easily identified in preparations such as canine liver, ear  and scrotal skin and rat tongue, no such cells were found in any species (except the cow) in any part of the vascular wall. The results indicate that in most species histamine is stored in the vascular wall in a non-mast cell pool which may well be in the smooth muscle cells themselves.


Failure of ellagic acid to affect platelet aggregation in normal and in factor XII deficient plasma.

Ellagic acid solutions, regardless of the concentration, failed to alter platelet aggregation induced by ADP, Adrenalin, Collagen, Thrombofax and Ristocetin in normal and in factor XII deficient plasma. A moderate inhibition was noted after addition of ADP both in normal and in factor XII deficient plasma but this was present also in the control systems and was therefore due to the buffer-dextrose solution and not to ellagic acid. These data indicate that Hageman factor plays on role in platelet aggregation.


A Swiss family with hemoglobin P Galveston beta117His leads to Arg, including two patients with hb P/beta thalassemia.

The mutant Hb P Galveston (beta117His leads to Arg) is observed in two heterozygotes for beta thalassemia and by itself does not cause clinical symptoms. Some of the physico-chemical properties of Hb P Galveston are identical to the onemical properties of Hb P Galveston are identical to the ones hemoglobin Zurich (beta 63 His leads to Arg) so that only a detailed analysis led to its proper identification.


Assessment of the value of prednisone test in differential diagnosis of neutropenic state.

The prednisone test revealed normal bone marrow reserve of neutrophils (BMR) in subjects with innocent neutropenias e.g. neutropenias not associated with  any other disturbances in haemopoiesis or with increased incidence of infections, assumed to be an individual anomaly of the subject. On the other hand, diminution or absence of MBR in chronic hypoplastic neutropenias, transient post-chemotherapeutic neutropenias and neutropenias with increased destruction of neutrophils were observed. The magnitude of that diminution was not dependent of the cause of neutropenia. It is suggested, that the prednisone test (and probably tests with other BMR mobilizing agents) may serve as screening tests for exclusion (or confirmation) of relative granulopoietic insufficiency as the cause of neutropenia. Only after confirmation of reduced BMR another explanations of neutropenia should be sought using other method as muramidase level, adrenaline test or granulocyte kinetics with DF32P, 3HDFP, or 51Cr.


Quantitative evaluation of the total number and distribution of lymphocytes in young pigs.

In young pigs, the spleen, thymus and all lymph nodes were dissected out and weighed. The relative content of lymphoid cells was determined from histological sections. The number of nucleated cells was evaluated by two different methods: firstly, by measuring the DNA content of samples of lymphoid tissue and dividing by the DNA content of a single nucleus; and, secondly, by counting all lymphoid cells in histological sections of defined volumes of these organs. The number of lymphoid cells in tonsils, gut, bone marrow and lung were determined using histological evaluations and the volumes or weights of these organs. The resulting average number of lymphocytes was 321 times 10 (9) for a pig of 26 kg body weight. The lymphocytes showed the following distribution in lymphoid and non-lymphoid organs: thymus 44%, spleen 9%, mesenteric lymph nodes 17%, cervical lymph nodes 9%, other peripheral lymph nodes 3%, gut-associated lymphocytes 5%, tonsils 2%, bone marrow 5%, blood 3%, lung 0.2% and an estimated figure of 3% for all other tissues.


[Stimulating effect of collagen and collagen derivatives on the propregation and adhesion of thrombocytes in defibrinogenated human citrate plasma and in animal citrate plasma].

Collagen type I form calf skin, collagen modified by pepsin, methylation, succinylation or deamidation and alpha1-chains or cyanogen bromide peptides were studied for their effect on adhesion and spreading of platelets. Some of the proteins increased platelet activity and indicated that the following structural parameters are of importance: 1. triple-helical conformation, 2. non-triple-helical regions, 3. charged amino acid side chains, 4. some activity was also detected for cyanogen bromide peptides of collagen alpha1-chain. The activity of collagen when used in a firbrinogen-free system is comparable to that of fibrinogen and potentiates platelet spreading caused by animal plasma.


A study of the flexor digitorium superficialis tendon in flexion and extension at the proximal interphalangeal joint.

Using a physical model to represent the detailed morphology of the flexor digitorium superficial tendon, the functions of the fibrous arrangement of this tendon in the human fingers were studied for flexion and extension of the proximal interphalangeal joint. In extension of the joint, the fibrous pattern acted as a dynamic ligament for the joint. In flexion, the arrangement of fiber bundles of the tendon contributed to the balanced distribution of forces across the joint's surfaces and for mechanical interplay with the deep long flexor tendon in the region of the joint.


Effects of desipramine on neuronal responses to dopamine, noradrenaline, 5-hydroxytryptamine and acetylcholine in the caudate nucleus of the rat.

1 The sensitivity of single neurones to microelectrophoretically applied dopamine, noradrenaline (NA), 5-hydroxytryptamine (5-HT) and acetylcholing (ACh) was investigated in the caudate nucleus of the rat, anaesthetized with halothane. Both excitatory and depressant responses could be observed to each of the agonists. There was a high correlation between the direction of responses to dopamine and noradrenaline, whereas there was no significant correlation between the direction of responses to dopamine and ACh. 2 The effect of desipramine was studied on both excitatory and depressant responses to dopamine, NA and 5-HT, and on excitatory responses to ACh. Both potentiation and antagonism of neuronal responses to monoamines and ACh could be observed after a brief application of desipramine. 3 Excitatory responses to glutamate were not affected by desipramine. 4 The observation that responses to dopamine and NA can be potentiated by desipramine in the caudate nucleus suggests that uptake blockade is not a prerequisite for potentiation. 5 It is suggested that the potentiation of neuronal responses to dopamine by desipramine may be responsible for the therapeutic efficacy of desipramine in Parkinson's disease.


Indomethacin-induced increase in noradrenaline turnover in some rat organs.

1 The effect of a prostaglandin synthesis inhibitor, indomethacin, on noradrenaline turnover rate in various rat tissues was determined from the product of the endogenous noradrenaline concentration and of the rate constant of (-)-[3H]-noradrenaline decline after injection of the labelled amine in tracer doses. 2 Treatment of the rats with indomethacin (5 mg/kg p.o. five times during 2.5 days) increased noradrenaline turnover rate 32-36% in submandibular gland, spleen and heart, and 4-17% in epididymal and subcutaneous adipose tissue. 3 The extrapolated [3H]-noradrenaline content at time 0 was greater in heart and adipose tissue of indomethacin-treated animals than in controls, while it was not significantly changed in spleen and salivary gland. 4 There was no apparent relationship between tissue concentrations of intravenously injected [14C]-indomethacin and the effect of indomethacin on noradrenaline uptake and turnover rate in the different tissues. 5 Indomethacin treatment did not affect monoamine oxidase and catechol O-methyl-transferase activities in the different tissues. 6 The results are consistent with the hypothesis that indomethacin increases noradrenaline turnover in the rat by blockade of a locally operating feed back inhibition of transmitter release by prostaglandins. However, additional effects, such as an increased impulse traffic, cannot be ruled out.


Inhibition of the pulmonary inactivation of prostaglandins in vivo by di-4-phloretin phosphate.

1 Inactivation of prostaglandin E2 in the pulmonary circulation of rabbits in vivo was measured by comparing the hypotensive effects of doses given intravenously and intra-arterially. 2 Di-4-phloretin phosphate (DPP) 25-100 mug kg-1 min-1 inhibited the inactivation of prostaglandin E2 in the pulmonary circulation. 3 These doses of DPP caused a marked shift to the left of the dose-response curve to prostaglandin E2 given intravenously but did not affect the dose-response curve to prostaglandin E2 given intra-arterially. 4 Inhibition of pulmonary inactivation of prostaglandins E2 and F2alpha caused marked potentiation of their gastrointestinal effects. 5 At these doses antagonism of the hypotensive action of prostaglandin E2 by DPP was seldom seen but the gastrointestinal effect of prostaglandin F2alpha was sometimes antagonized. 6 After treatment with DPP 100 mug kg-1 min-1, high doses of prostaglandin were sometimes lethal.


Potency and selectivity of methyl analogues of prostaglandin E2 on rat gastrointestinal function.

1 The potency and selectivity of action of prostaglandin E2 and its (15S)- or (15R)-15 methyl and 16, 16 dimethyl analogues on gastrointestinal function have been studied in the rat. 2 The (15S)-15 methyl and 16, 16 dimethyl analogues were 40 times as active as prostaglandin E2 in inhibiting pentagastrin-stimulated acid secretion on intravenous administration to the anaesthetized rat, and 100 times as active on subcutaneous injection to the chronic fistula rat. 3 In antisecretory doses, the analogues, like prostaglandin E2, caused bile reflux and, in higher doses, profuse diarrhoea. 4 The (15S)-15 methyl and 16, 16 dimethyl analogues were at least 30 times as active as prostaglandin E2 in causing changes in intestinal intraluminal pressure in vivo, but were equipotent on isolated smooth muscle. 5 In equivalent antisecretory doses, the methyl analogues had little effect on systemic arterial blood pressure and resting mucosal blood flow compared with prostaglandin E2. 6 The (15R) methyl epimer administered parenterally had little effect on gastrointestinal function but brief acid incubation greatly increased its activity.


An analysis of the depressor responses to histamine in the cat and dog: involvement of both H1- and H2-receptors.

1 The depressor responses to histamine, in anaesthetized cats and dogs, have been shown to involve both histamine H1- and H2-receptors. 2 In both species, histamine caused dose-dependent falls in blood pressure. The dose-response curve could be displaced to the right by administration of mepyramine 2.5 times 10(-6) mol/kg i.v. The displacement was maximal with a dose-ratio of less than ten. 3 Metiamide alone, up to 2 times 10(-6) mol kg-1 min-1, had no significant effect on the histamine dose-response curve. 4 When administered in the presence of mepyramine, metiamide, 4 times 10(-7) and 2 times 10(-6) mol kg-1 min-1, caused dose-dependent displacements to the right, of the histamine dose-response curve greater than could be achieved with mepyramine alone. 5 The results indicate the presence of both histamine H1- and H2-receptors in the cardiovascular system of the cat and dog. Both receptors produce a common response, i.e. a fall in blood pressure.


Innervation both of peri-orbital structures and of the heart by the cervical sympathetic nerves in mouse, rat, guinea-pig, rabbit and cat.

1 In anaesthetized rats electrical stimulation of the intact cervical sympathetic nerve produced frequency-dependent lower eyelid contractions and tachycardia. 2 The tachycardia was caused by excitation of efferent fibres since it was equally evident in the pithed rat preparation, and the right nerve was more effective than the left. By contrast, no differences were seen between the responses to right and left vagal stimulation in either rats or rabbits. 3 Guanethidine inhibited both cardiac and eyelid responses, propranolol only the former and phentolamine only the latter, therby revealing the adrenergic nature of the nerves. Hexamethonium caused partial inhibition and the block was intensified by atropine. 4 The inferior eyelid of mice, guinea-pigs and rabbits as well as the nictitating membrane of rabbits and cats were contracted by cervical sympathetic nerve stimulation. In these species too, tachycardia occurred; this was more pronounced with the right than the left sympathetic nerve. The order of cardiac responsiveness was mouse greater than rat greater than guinea-pig greater than rabbit greater than cat. 5 In guinea-pigs histamine-induced bronchoconstriction was reduced by cervical sympathetic nerve stimulation. 6 That discrete cardiac pathways exist in the cervical sympathetic nerves is suggested by the reproducibility of the effects within any one species. The accessibility of the nerves greatly simplifies the examination of drugs in vivo on two different structures innervated by the sympathetic nervous system.


Decamethonium in the perfused and immersed rat diaphragm.

1 The water content and mannitol space of rat diaphragms which were perfused through the inferior vena cava was increased compared with immersed diaphragms. The potassium content of both preparations, when expressed in terms of dry weight, was maintained at similar values to that found in vivo. 2 Despite the application of a constant concentration of decamethonium, a steady level of neuromuscular block was not obtained in either the perfused or immersed rat diaphragm. The immersed preparation differed from the perfused preparation in that recovery from paralysis occurred despite the continued presence of the drug. 3 The rate of uptake of labelled decamethonium (100 muM) at the end-plate region was similar in the perfused and immersed diaphragm. The slopes of the regressions were 0.059 and 0.054 mul mg-1 min-1 (based on dry weight) respectively which were not significantly different. This implies that the rate of uptake of the drug at the end-plate is slow and limited by the rate of entry into the fibre rather than by diffusion to the site of entry.


Rapid effects of environmental disturbance on rat plasma unesterified fatty acid and tryptophan concentrations and their prevention by antilopolytic drugs.

1 Changes of plasma unesterified fatty acid (UFA) and tryptophan concentration in group-housed rats following removal of their cage-mates and the effects of antilipolytic drugs on these changes were investigated. 2 Removal of group-housed 24 h fasted rats but not fed rats from cages resulted in increased plasma UFA concentration in the remaining rats which was associated with significant increases of the proportion of free tryptophan but significant falls of total tryptophan concentration. These rapid changes were not associated with brain tryptophan changes. Plasma tyrosine concentration was unaffected. 3 The fall of plasma tryptophan did not appear to be due to passage into red cells as erythrocyte tryptophan concentration remained unchanged. 4 Plasma UFA concentrations correlated positively and significantly with corticosterone concentrations which were also increased following removal of cage-mates. 5 Plasma UFA increases and tryptophan changes in the fasting rats were both prevented by nicotinic acid or propranolol. Corticosterone concentration was increased by nicotinic acid but unaffected by propranolol. 6 The possible importance of these rapid changes of plasma tryptophan and of their prevention by antilipolytic drugs is discussed.


Comparative bioassay of prostaglandin E2 and its three pulmonary metabolites.

The relative potencies of prostaglandin E2 and its metabolites 15-keto PGE2, 13,14-dihydro-15-keto-PGE2 and 13,14,dihydro-PGE2 were investigated on isolated smooth muscle preparations. These were rat stomach strip, colon and uterus, chick rectum, guinea-pig ileum, trachea and pulmonary artery and rabbit aorta and pulmonary artery. 15-keto PGE2 was equiactive or 1-1.8 times more potent than prostaglandin E2 in relaxing guinea-pig trachea but otherwise the three metabolites were less active than prostaglandin E2 on all preparations.


In vitro inhibition of brain mitochondrial monoamine oxidase by 6-hydroxydopamine.

6-Hydroxydopamine (6-OHDA) inhibits rat brain mitochondrial monoamine oxidase (MAO) when kynuramine or dopamine are used as substrates. The effect is competitive and reversible giving Ki values of 74 and 176 muM with the respective substrates. At high concentrations (5 mM) of each substrate, inhibition of MAO was not observed.


Study of coronary risk factors related to physical activity in 15 171 men.

Physical activity, both at work and during leisure, was assessed in 15 171 men aged 25-74 years. Heavy leisure activity was associated with lower mean serum cholesterol levels and blood pressure. In men under 60 years the same negative association was also noted between leisure activity and relative weight and cigarette smoking. Different degrees of physical activity at work were not associated with any differences in these risk factors, nor did the level of exercise at work seem to influence the negative association between leisure activity and risk factors. Heavy leisure activity in young and middle-aged men is associated with lower levels of certain coronary risk factors and, therefore, a lower risk of coronary heart disease. The often-reported reduction in coronary morbidity and mortality with physical exercise may not be the direct effect of the exercise itself.


Thalassaemia, iron, and pregnancy.

Haematological values of 35 pregnant women with beta-thalassaemia trait were followed during pregnancy. The discriminant function, calculated from haematological indices, was of no value in diagnosing beta-thalassaemia trait during pregnancy. Initially patients were given iron supplements only if the serum iron and total iron binding capacity levels indicated iron deficiency, but bone marrow biopsies performed in the first 22 patients at 32 weeks indicated deficient iron stores. These patients were therefore given iron irrespective of their serum iron level. All subsequent patients with beta-thalassaemia were also put on iron routinely at booking. Retrospectively the patients were divided into two groups. Patients in group 1 (18 patients) had received iron for less than 12 weeks, and their haemoglobin levels fell significantly during pregnancy (P less than 0-001). Haemoglobin levels in 16 patients who had received iron for more than 12 weeks (group 2), however, did not fall significantly during pregnancy (P less than 0-6). It is suggested (contrary to common practice) that patients with beta-thalassaemia trait should be given iron supplements during pregnancy. Serum folate and vitamin B12 levels did not change significantly in these patients and there was no increase in the incidence of maternal or fetal complications.


Evaluation of creatinine phosphokinase in screening patients for malignant hyperpyrexia.

Evidence is presented that serum creatinine phosphokinase (CPK) activity is of no direct value in screening patients for susceptibility to malignant hyperpyrexia and does not correlate with halothane-induced muscle contracture or the presence of myopathy. Widely differing CPK values were found at different times in the same people. In most "malignant hyperpyrexia" families the susceptible patients had either normal or inconsistently raised CPK values.


Cerebrospinal fluid gamma aminobutyric acid levels in migraine.

Gamma-Aminobutyric acid (GABA) levels in cerebrospinal fluid were measured in seven patients with tension headache and 12 patients with migraine. GABA was detected only during the migraine attack. The results suggest disordered GABA metabolism in migraine.


Self-inflicted injury: a follow-up study of 43 patients.

Forty-three patients, 38 women and 5 men, with self-inflicted skin lesions were studied. Thirty-three were followed up for up to 22 years. In most cases dermatitis artefacta was only one incident in a long history of psychogenic illness. Of the 43 patients, 13 (30%), 12 women and one man, continued to produce lesions or were disabled with other psychiatric disorders more than 12 years after the onset of symptoms. Prognosis was difficult but recovery seemed to occur when the patient's life circumstances changed rather than as a result of treatment.


Isolated glomerulonephritis with mesangial IgA deposits.

Mesangial deposits of IgA, occurring in the absence of systemic disease known to be associated with nephritis, were detected by immunofluorescence microscopy in renal biopsy specimens from 25 patients (4% of 630 specimens studied). Associated deposits of C3 were always present, usually with IgG, but IgM deposits were less common and C1q was never seen. On light microscopy most of the biopsy specimens showed mesangial of focal nuclear proliferation though some were normal. Fifteen of the 25 patients presented with macroscopic haematuria, which was usually recurrent and preceded by a sore throat, whereas the remaining, and usually older, patients presented with persistent proteinuria and were more likely to have impaired renal function. This incidence of "mesangial IgA disease" is less than that reported by French workers. There was a significantly high incidence of familial renal disease among these patients. No abnormalities of serum complement or IgA concentration were found.


Erythropoietic uroporphyria of Gunther first presenting at 58 years with positive family studies.

Erythropoietic uroporphyria of Gunther was seen in a 58-year-old man who presented with photosensitivity, haemolytic anaemia, and classical laboratory findings. Family studies showed five asymptomatic relatives with erythrocyte uroporphyrin concentrations in the probable latent heterozygote range.


Clobetasone butyrate, a new topical corticosteroid: clinical activity and effects on pituitary-adrenal axis function and model of epidermal atrophy.

Clobetasone butyrate is a new corticosteroid, selected for study because of its combination of good activity in the vasoconstriction test and low systemic activity in animals. Formulated as an 0.05% ointment and cream (Molivate) it was clinically effective in patients with eczema, its activity being significantly greater than that of hydrocortisone 1% or fluocortolone 0.2% (Ultradil). Under conditions that predispose to maximal percutaneous absorption clobetasone butyrate ointment had minimal effect on hypothalamic-pituitary-adrenal function. In an animal model of cutaneous atrophy it caused less thinning of the epidermis than steroids other than hydrocortisone. Clobetasone butyrate 0.05% ointment and cream gave every indication of offering clinically effective topical anti-inflammatory activity with a wide margin of safety.


The motor units of cat medial gastrocnemius: speed-size relations and their significance for the recruitment order of mortor units.

The interrelationships between axonal conduction velocity, tetanic tension, twitch contraction time and rate of force development during a tetanus have been studied in 126 motor units from 12 cat medial gastrocnemius muscles. While the range of axonal conduction velocities for slow- (contraction time greater than 45 msec) and fast-twitch units overlapped, the mean conduction velocity for the slow-twitch group was significantly lower. No difference could be found between the axonal conduction velocities of the fast non-fatiguing and fast fatiguing units. Within individual experiments, few significant correlations were found between conduction velocity and tetanic tension or contraction time for the fast- and slow-twitch units. Some correlations did appear when data from these populations were pooled, but such results are shown to be misleading. Some weak correlations were found between motor unit contraction strength and twitch contraction time. The rate of rise of isometric force development was found to be most strongly related to tetanic tension and only weakly related to contraction time. The ordering of motor units according to contraction strength reveals the association of motor unit mechnical properties to be ideally suited for the dual role of medial gastrocnemius as a postural and powerful phasic muscle.


Ageing and dopamine uptake by subcellular fractions of the C57BL/6J male mouse brain.

The uptake of dopamine (DA) by synaptosomes from the hypothalamus and striatum was reduced in healthy ageing mice at DA concentrations below 10(-7) M. Kinectic analysis indicates that ageing increases the Km of high affinity DA uptake processes by about 35%, but does not alter alter Vmax. l-Tyrosine and serotonin uptake at low concentrations was unaltered by ageing in the striatum and hypothalamus; no changes of L-norepinephrine uptake were found in the hypothalamus. We conclude that ageing may selectively affect DA uptake processes by neuronal membranes.


Coordination of movements of the kindlimbs and forelimbs in different forms of locomotion in normal and decerebrate cats.

The coupling of movements of the hindlimbs and forelimbs has been analysed in intact cats stepping overground and on a treadmill and during swimming, and in decerebrate cats stepping on a treadmill, immersed in water('swimming') and stepping suspended in the air. In the different preparations, and under different types of locomotion, two basic patterns of coupling have been observed. Both concern the hindlimb and forelimb of the same side of the body. The first pattern is found in the pacing gait where flexion of the forelimb precedes extension of the hindlimb, measured at the elbow and knee, respectively. The second pattern is typically found in the trot where flexion of the forelimb follows extension of the hindlimb. In decerebrate cats both patterns of coupling remain after bilateral deafferentation of the hindlimbs. In the alternate form of locomotion these patterns of coupling occurs symmetrically on both sides. In the rotatory and transverse gallop (examples of the in-phase form of locomotion) the coupling is asymmetrical: on one side it is comparable to pacing (forelimb flexion precedes hindlimb extension), and on the other side to trotting (forelimb flexion follows extension). These basic patterns of interlimb coordination simplify considerably the problem of neural control of the limbs in locomotion. Obersations of EMGs during the alternative forms of locomotion show that in the pacing type of coupling the extensor EMGs of forelimb and hindlimb overlap, with the hindlimb leading the forelimb by about 10% of a step cycle, while in the trotting type of coupling the forelimb flexor EMGs overlap the hindlimb extensor EMGs, the forelimb flexors leading the hindlimb extensors by about 10% of a step cycle. During acceleration the transition between the two forms of EMG occurs within one or two step cycles, and at some intermediate velocities the EMG coupling springs back and forth between the two different forms. These results further support the hypotesis of two basic forms of interlimb coupling in which long propriospinal pathways probably play a role.


Movements of the forelimbs of the cat during stepping on a treadmill.

In normal cats stepping on a treadmill an analysis has been made of movements of the scapula, shoulder, elbow, wrist and digits. The scapula is capable of making large and complicated movements over the rib cage. In locomotion they may be resolved into a cyclical, similunar movement of the glenoid cavity in the parasagittal plane of the cat's body. The movements of the scapula are rather constant over a wide range of velocities. They most resemble those of the hip which also shows a delay in the onset of extension and no yield phase. It is suggested that movements of the scapula have a high priority in the execution of locomotion in the forelimbs. The movements at the shoulder tend to parallel those of the elbow. The terms of palmar flexion and dorsiflexion are retained for the movements at the wrist and digits. The rapid phase of palmar flexion corresponds with the flexion phase at the elbow, and the phase of dorsiflexion with the first extension phase at the elbow. Palmar flexion during the second and third extension phases of the more proximal joints would appear to be initiated by contact of the foot with the ground. The sequences of movement at the wrist and digits allow a comparison of the roles of flexor and extensor muscles of the forearm during locomotion and the flexion reflex, which is compatible with the functional and anatomical organization of cervicothoracic segments. With exception of extension at the scapula the movements of the elbow provide an index of the onsets of flexion and extension at the other joints of the forelimb. Modifications are suggested for the Phillippson step cycle of both forelimbs and hindlimbs to include the delayed onset of extension at the scapula and hip and the particular movements of the wrist, digits and toes.


Genetic recombination in a chloramphenicol-producing strain of Streptomyces species 3022a.

Mutation with ultraviolet light and chemical mutagens yielded strains with auxotrophic and streptomycin-resistance markers. These were used to show the existence of genetic recombination in Streptomyces species 3022a and to construct a linkage map for 17 marker loci.


Interactions between Azotobacter and "phosphobacteria" and their establishment in the rhizosphere as affected by soil fertility.

The effects on plant growth of "bacterial fertilizers" prepared from Azotobacter spp. and phosphate-solubilizing bacteria ("phosphobacteria") have been the subject of much controversy. Cases where no plant-growth stimulation occurred may often be accounted for by the failure to establish the bacterial inocula in the rhizosphere. Three factors that may influence inocula establishment, i.e. soil fertility, manuring, and interactions between Azotobacter and "phosphobacteria," were examined in pot experiments, designed for statistical analysis, in two neutral-alkaline soils, using lavender plants (Lavandula spica L.). During the experiments the numbers of Azotobacter and "phosphobacteria" were counted. Dry weights of roots and shoots were recorded after 16 weeks of growth. At the end of the experiments there were always more Azotobacter and "phosphobacteria" in the rhizospheres when plants were inoculated with both groups of organisms together than when they were inoculated singly. Addition of 2% farmyard manure to the richer soil enhanced this effect. Plant growth was greatest when seedlings were inoculated with both Azotobacter and the "phosphobacteria".


Nutritional requirements of some non-pathogenic Neisseria grown in simple synthetic media.

Ten species of non-pathogenic Neisseria were grown in simple defined liquid media containing amino acids, biotin, nicotinic acid, calcium pantothenate, ferrous sulfate, magnesium sulfate, and potassium phosphate. Two of these Neisseria were induced to grow with glutamic acid as the carbon and nitrogen source. The remaining eight Neisseria grew in glutamic acid medium supplemented with from one to four additional amino acids, lactate, or lactate and glucose. A strain of N. flavescens grew in the absence of added growth factors whereas the remaining nine species of Neisseria required either biotin or nicotinic acid; pantothenate was required by two and was stimulatory for three of these species. Use of carbohydrates by the non-pathogenic Neisseria in synthetic medium was tested. Two strains failed to use any of the 14 carbohydrates tested; one strain used only glucose; the remaining seven strains used fructose, glucose, maltose, and sucrose to varying degrees.


Soil bacteriostasis: inhibition of spore germination and microcolony development in agar discs incubated on nonsterile soils.

An agar-disc method showed that bacterial spores were subject to a bacteriostatic effect exerted by soil. Soil either inhibited spore germination or reduced microcolony development of the six Bacillus strains tested. Bacteriostasis was relieved by the addition of nutrients to soil-exposed discs after they had been removed from the soil.


Characterization of the factors responsible for death of fish infected with Vibrio anguillarum.

Vibrio anguillarum produced substances toxic for goldfish (Crassius auretus) that are released from living bacteria and associated with heat-killed bacteria. Heating (100 degrees C) enhances the potency of the extracellular toxin.


[Family psychiatry, transactional systems, and the frontiers of child psychiatry].

Family therapists, seen as emphasizing interventions leading to change in family structure and in sequences of behaviour, have been searching for an integrated concept which transactional systems theory seems to offer. It takes into account small group theory, social role theory, communications theory, and general system theory, which are linked to psychic, somatic, socio-cultural, politico-economic, and ecological factors. This idea of interlinked, open systems which influence each other is used as a viewpoint for examining the frontiers of child psychiatry. Adult patients often have children who are affected by their parents' treatment, and child psychiatrists often intervene with adults. Similarly, the boundaries between psychiatry and the para-medical professions have grown less distinct as we have become aware of more elements to assess in each case, and as the number of therapeutic techniques and possible interventions increase. The problem of defining child psychiatry is discussed, as is psychiatric training, in terms of the difficulty in integrating the many theoretical and practical levels.


Ultrastructure and fluorescence histochemistry of endocrine (APUD-type) cells in tracheal mucosa of human and various animal species.

This study describes distrinctive cells with ultrastructural and histochemical features of APUD-type endocrine cells within the tracheal epithelium of human fetuses, newborns and children as well as different animal species. These cells referred to as Kultschitzky cells (K cells) were found to be argyrophilic, but not argentaffin, and are considered analogous to the same type of cells in lung and gastro-intestinal tract. Fluorescence histochemistry demonstrated the presence of intracellular amine within tracheal K cells, but only after in-vitro or in-vivo administration of amine precursor (L-DOPA). Ultrastructurally, these cells are characterized by the presence of numerous cytoplasmic granules (dense core vesicles) which show species related morphologic variations. Two different types of K cells were found in trachea of lamb and armadillo, each type possessing morphologically different dense core vesicles. In human and rabbit tracheas, only one type of K cell was identified. K cells in the trachea are distributed as single cells between other epithelial cells; neuroepithelial bodies such as those found in bronchial mucosa were not identified. Well differentiated K cells were found in tracheas of early human fetuses and throughout gestation, infancy, and childhood. Preservation of K cells in human autopsy material and widespread occurence of these cells in various laboratory animals will permit further studies into the nature and function of tracheobronchial endocrine cells.


The kinetics of 35S-labelled cysteine in the hypothalamic-neurohypophyseal neurosecretory system of the dehydrated rat.

Thirst stimulation of the supraoptic nucleus (SON) and paraventricular nucleus (PVN) was induced in rats by withholding all fluids during three days. 35S-cysteine was then intraperitoneally administered and the rats were killed at predetermined times and examined by autoradiography, applying the authors' previously described method. This experimental series totalling 51 animals was compared with a control series of 70 rats, similarly treated, who had had free access to water. The kinetic phenomena in SON and PVN were analysed in terms of the two-compartment model previously used, which gives an estimate of the neurosecretory material (NSM) secretion parameters and of those of the lumped structural cell protein turnover in the nuclei. The kinetics of the precursor amino acids after administration of labelled cysteine were also assessed. Determinations of the label uptake at two specific times in the experiment, in the infundibular nucleus, ventromedial nucleus and optic nerve tissue in both series served as a check on the specifity of the structural protein turnover changes observed. Compared with the controls, the turnover rate of the slow compartment was more than tripled in the dehydrated rats, while that of the fast compartment had gone down to about one-third; both effects very nearly equal in SON and PVN. These results are compatible with the concept according to which thirst stimulates the SON and PVN equally. A distinct, and strikingly equal, hump was observed (2 hours after label administration) in all specific activity curves, also in the precursor serum concentration, and it is probably due to recycling of 35s from cysteine to methionine. This and other circumstances render the phenomena rather too complex for a straight-forward evaluation by the two-compartment model. Even so, the observations are believed to furnish good evidence of the biological verity of this model as well as the thirst-induced changes elicited.


Presence of a pineal nerve in sheep and rabbit fetuses.

The presence of a nerve located just caudal to the pineal gland in the midsagittal plane is demonstrated in sheep and rabbit fetuses. This nerve lies freely in the subarachnoid space and extends from the pineal gland to a region of the CNS located dorsal to the rostralmost part of the subcommissural organ (SCO). In rabbit fetuses the nerve is obsered on days 23 and 24 of gestation; we suggest that it is an ontogenetic equivalent to the pineal nerve of anuran amphibians. The developmental fate of the mammalian fetal pineal nerve is dicussed.


Fine structure of the gastric epithelium of the ascidian Botryllus schlosseri. Mucous, endocrine and plicated cells.

The following five cell types have been recognized and defined on the basis of their fine structure in the gastric epithelium of B. schlosseri: vacuolated and zymogenic cells (described in a previous paper); ciliated mucous, endocrine and plicated cells. The ciliated mucous cells are distributed at the apex and the bottom of the gastric folds and along the dorsal groove. The mucus droplets appear to form from the Golgi complex as secretory granules of variable density and texture, which are released from the cell after fusion of their membranes with the apical plasma membrane. Holocrine or apocrine secretion has not been observed. The endocrine cells are scattered and are characterized by electron dense granules, especially numerous in the basal region of the cell. Finally, the plicated cells, present in the pyloric caecum, show rod-like microvilli, a well developed Golgi complex and abundant, deep infoldings of the basal plasma membrane, which are associated with numerous mitochondria. The possible role of the gastric cell types is discussed taking into account information concerning morphologically similar cells in other animals, as well as previously reported data on the biochemistry and physiology of digestion and excretion in ascidians.


Prolactin and growth hormone cells in the human hypophysis: a study with immunoenzyme histochemistry and differential staining.

Growth hormone and prolactin cells were immunostained in human hypophyses with antibody against rat growth hormone or prolactin and the peroxidase-antiperoxidase complex. Growth hormone cells were round and, in normal pituitaries, arranged in sizable groups. Prolactin cells occurred singly and were less numerous; they were often extensively branched. Only a few prolactin cells stained with carmoisine. Incubation of the antibody with an excess of the appropriate antigen greatly diminished or abolished immunostaining; absorption of anti-prolactin with growth hormone often enhanced it. Prolactin cells were somewhat hypertrophied and hyperplastic in a neonate. Many of them stained with carmoisine. An even greater hypertrophy and hyperplasia of these cells (which pushed apart the growth hormone cells) was found in a lactating woman. Immunostained giant prolactin cells were also observed. Staining of the prolactin cells with carmoisine was extensive. Upon prolonged exposure to anti-growth hormone antibody, ACTH/MSH cells also showed immunostaining which was abolished by absorption of the antiserum with growth hormone but not with synthetic 1-24ACTH. Growth hormone cells evidently correspond to the alpha acidophils of Romeis, prolactin cells in lactation to his eta cells; the relation of his epsilon cells to the pleomorphic "resting" prolactin cells is not clear.


Identification of separate vasopressin-neurophysin II and oxytocin-neurophysin I containing nerve fibres in the external region of the bovine median eminence.

Immuno-enzyme histochemical investigations showed the presence, in the external region of the bovine median eminence, of accumulations of vasopressin-neurophysin II- and oxytocin-neurophysin I-complexes. These two hormone-neurophysin complexes are located in separate fine varicose nerve fibres. The results strongly plead against an important role of tanycytes in the transport of vasopressin, oxytocin and neurophysins from the cerebrospinal fluid to the hypophysial portal blood-vessels.