Sclerosis in non-Hodgkin's lymphomata.

Sclerosis in non-Hodgkin's lymphomata. A total of 242 cases of non-Hodgkin's lymphoma arising primarily in lymph nodes are reported. Lymph nodes have been classified histologically and the incidence of sclerosis in each group assessed. Two types of fibrosis were found, one composed of fibrous bands from 10 mum to several hundred mum thick within the node and invaded perinodal tissue. The other type was a fine compartmentalizing fibrosis sometimes simulating the pattern of metastatic carcinoma in part of a node, the rest of the node being typical of malignant lymphoma. Both types of fibrosis were present in some nodes. Fibrous bands were present in all histological types, with the exception of the diffuse lymphocytic well differentiated group. Fine compartmentalizing fibrosis was seen only in association with areas of undifferentiated large celled lymphoma. The presence of sclerosis is shown to correlate with a better prognosis when compared with non-sclerosing tymours of similar histological type. That sclerosing lymphomata are more slowly progressive is borne out by the fact that two-thirds of the patients with sclerotic nodes presented with localized disease compared with less than half of the patients whose nodes showed no sclerosis.


Clinical trials in the non-Hodgkin's lymphomata at Stanford University experimental design and preliminary results. Preliminary results of controlled clinical trials in 127 patients with non-Hodgkin's lymphomata are reported. The rationale for identifying 7 study groups is presented. Treatment programmes which are being studied vary from involved field irradiation to the most aggressive treatment tolerable. The early results suggest that patients with both favourable and unfavourable histologies but of limited extent can be controlled with irradiation. Patients with advanced disease but with favourable histological subgroups are doing as well with a conservative drug programme as with 2 aggressive treatment regimens. These controlled studies and those of others, facilitated by the use of the Rappaport classification and laparotomy staging, give promise of clarifying and improving the management of patients with these lymphomata.


Results of combination chemotherapy of non-Hodgkin's lymphoma. The analysis of the results of CVP and MOPP chemotherapy in 80 patients with advanced stages of non-Hodgkin's lymphoma shows that 36 achieved a complete remission. Twenty-five percent of all patients remain free of disease for periods ranging from 4 months to over 7 years, with a projected median duration of complete remissions of 3 1/2 years. Well differentiated and nodular histology were positive determinants for survival, confirming the overall clinical validity of the Rappaport classification system for the non-Hodgkin's lymphomata. However, it has been demonstrated that it is possible to achieve a complete remission using combination chemotherapy even in the most clinically aggressive histological subgroups, and that these responses can be correlated with an extended survival. An analysis of patterns of relapse from complete remission in patients with non-Hodgkin's lymphoma treated with combination chemotherapy has demonstrated differences between histological subgroups. Patients with diffuse histiocytic lymphoma who achieved a complete remission after 6 months of treatment without maintenance have remained disease-free, whereas those with nodular poorly differentiated lymphocytic and diffuse well differentiated lymphocytic lymphomata demonstrate a pattern of continuous late recurrence. The initial sites of relapse from complete remission in lymphocytic lymphoma were lymph nodes and bone marrow which were involved before treatment.


Therapy of advanced lymphocytic lymphoma a preliminary report of a randomized trial between combination chemotherapy (CVP) and intensive radiotherapy. The initial results of a randomized clinical trial comparing intensive cyclical combination chemotherapy (CVP) and total body radiation therapy in the treatment of advanced (Stage III and IV) lymphocytic lymphoma is presented. Sixty-five patients have been entered and randomized according to stage. Of the chemotherapy treated patients, (22/27 (81%) achieved a response with 55% complete responders. The radiation group or those treated with radiation 27/32 (84%) responded with 56% in complete response. There is no significant difference in overall survival between the two groups. Nodular lymphoma (44 patients) responds more often and has s significantly longer survival than diffuse lymphoma (21 patients). Approximately 50% of the complete remitters have relapsed but reinduction of disease control was possible in almost all cases.


The influence of chemotherapy on the management of non-Hodgkin's lymphomata at the Princess Margaret Hospital. A comparison of the results from 1962-64 with 1967-69. A retrospective study of 460 non-Hodgkin's lymphoma patients seen in 1962-64 (185) and 1967-69 (275) revealed a marked improvement in the survival of patients treated during the later interval. The increased survival was not due to differences in the age or sex distribution of patients, the proportions with systemic symptoms or extranodal presentations. The improvement was due partially to a disproportionate increase in the number of patients with well- and intermediately differentiated nodular lymphocytic lymphomata in 1967-69. When these patients  were removed, the proportions of patients with "good" and "poor" pathology were comparable for the two intervals, and a markedly improved survival was still evident for the 1967-69 group. An improvement in the proportion of patients achieving a complete remission occurred only in patients who received some form of chemotherapy. Relapses occurred in 44.6% of patients with Stage I and II disease treated with radiotherapy alone. The results of this study indicate that we must re-evaluate carefully the indications for radiotherapy in the treatment of the non-Hodgkin's lymphomata. The addition of chemotherapy appears to be a promising method for increasing the proportion of patients who will achieve a long complete remission.


Cytology in the classification of diffuse non-leukaemic malignant lymphomata (lympho- and reticulosarcomata). Cytological examination of smears or imprints of diffuse non-leukaemic lymphomata gives more details of the morphological aspects of the cells than does histological examination. It enables us to distinguish (a) prolymphocytic (but not lymphocytic), (b) lymphoblastic or lymphoblastoid and immunoblastic lymphosarcomata. It helps to diagnose so-called reticulosarcomata from carcinomata and to distinguish two types: (a) cytic and (b) blastic, but it makes us suspicious about the nature of the latter. Light microscopy shows the cells of the blastic type resemble transformed lymphocytes (immunoblasts) more than reticulum cells. Electron microscopy shows many polyribosomes, which enhances the suspicion that some so-called "reticulosarcomata" could be immunoblastic lymphosarcomata.


Follicular lymphoma: A survey of 75 cases with special reference to the syndrome resembling chronic lymphocytic leukaemia. In a series of 75 cases of follicular lymphoma collected between 1950 and 1971 the following features were singled out as characteristic of, or specially frequent in, follicular lymphoma: (1) the high incidence in females; (2) the long history of symptomless lymph node enlargement; (3) the frequent presence of extensive symptomless disease at presentation, especially in patients below 40 years of age; (4) the syndrome resembling and often confused with chronic lymphocytic leukaemia, in which the notched nucleus cell, when present, is diagnostic; (5) the responsiveness of extensive disease to radiotherapy and alkylating agents at low dosage; (6) the long overall survival and the long survival after relapse; (7) the occurrence of abrupt change in the character of the disease in which, during a period of apparent quiescence, malignant transformation supervenes proving rapidly fatal. In the transformed stage the disease behaves as an invasive metastasizing sarcoma or as a blast cell leukaemia; in both cases the transformation is associated with the appearance of a new cell type, undifferentiated and distinct from that present before onset of transformation, which may however persist. The syndrome resembling chronic lymphocytic leukaemia occurs only in follicular lymphoma.


Electron microscopy in the non-Hodgkin's lymphomata. The component cells of peripheral lymphoid tissue have been divided into the lymphocyte and plasma cell lines, mononuclear phagocytic cells, dendritic "reticular cells", the reticular (supporting) cells and endothelial cells, and it is suggested that this system of cells should collectively be referred to as the lymphoreticular monoclear phagocyte system or LRMPS. Seventeen tumours of the LRMPS (excluding Hodgkin's disease) have been studied at ultrastructural level. Of these 17 non-Hodgkin lymphomata 5 were follicular lymphomata and 12 diffuse. It is concluded that electron microscopy plays a valuable role in the diagnosis of this group of tumours. Not only does it allow rejection of a diagnosis of lymphoma in certain anaplastic tumours, but it also enables a more precise identification of the cellular components of a lymphoma as well as indicating the degree of differentiation of the cell line involved. Additional advantages are the visualization of subcellular structures useful as markers, and by means of specialized immunoelectron microscopic techniques the identification of antigens and antibody formation within a given tumour. Two other results of this ultrastructural study are the indication that the dendritic cells of lymphoid follicles are derived from capillary endothelium, and the identification of certain anomalous formations derived from rough endoplasmic reticulum in the case of tumours showing plasmacytoid differentiation.


Leukaemic conversion of non-Hodgkin's malignant lymphomata. In 143 patients with poorly differentiated lymphosarcoma, leukaemic conversion has been observed in 25. The cytological type was prolymphocytic or lymphoblastic or lymphoblastoid (immunoblastic ?). Twenty-five patients were treated with chemo-radiotherapy, followed by active immunotherapy as if they had primary acute lymphoid leukaemia. A complete remission was obtained in 11. Four are still in first complete remission after 4 1/2 years. Among 136 patients suffering from so-called "poorly differentiated reticulosarcoma", 17 became leukaemic. The cells are cytologically very dystrophic and unidentifiable. A remission was obtained in 7 patients but it was of short duration (median 1 1/2 months, longest 7 months).


Humoral and cellular immunity in atopic eczema. Parameters of humoral and cellular immunity were measured in thirty-five patients with atopic eczema. The mean serum IgE level was raised but levels of the other major immunoglobulin classes were normal. Ten per cent of patients failed to respond to tetanus immunization. All patients responded to S. typhi H antigen. Fourteen per cent of patients failed to mount delayed hypersensitivity reactions to a battery of three intradermal antigens. The phytohaemagglutinin-stimulated uptake of 3H thymidine by lymphocytes was normal in the presence of autologous or of fetal calf serum, as was the spontaneous lymphocyte uptake. T and B lymphocyte numbers in the peripheral blood were normal. These results are similar to those found in asthmatic patients and support the hypothesis that, in some patients, atopic eczema is associated with an immunodeficiency state.


Viral warts, herpes simplex and herpes zoster in patients with secondary immune deficiencies and neoplasms. The incidence of viral warts, recurrent herpes simplex and herpes zoster has been assessed in a group of patients with possible secondary immune deficiency states and compared to a control group. Patients with cell-mediated immune deficiency appear to be more prone to warts and zoster infection as compared to patients with humoral immune deficiency. Recurrent herpes simplex infection was not increased in either group.


Ultrasonic measurement of fetal abdomen circumference in the estimation of fetal weight. A method of estimating fetal weight by ultrasonic measurement of the fetal abdominal circumference is descirbed. Assessment of birth weight predictions on 140 fetuses who were delivered within 48 hours of this measurement showed that the accuracy of predictions varied with the size of the fetus; at a predicted weight of 1 kg, 95 per cent of birth weights fell within 160 g, while at 2 kg, 3 kg and 4 kg the corresponding values were 290 g, 450 g and 590 g respectively. Expressed as a percentage of the predicted weight, confidence limits remained constant throughout the birth weight range. Extrapolation of these data to routine screening of the obstetric population showed that with a single measurement at 32 weeks menstrual age, 87 per cent of babies below the 5th centile would be detected by this method but that the diagnosis rate would fall to 63 per cent at 38 weeks. The false positive diagnosis rate would remain constant between 32 and 38 weeks at just over 1 per cent.


Comparison of intravenous oxytocin and prostaglandin E2 for induction of labour using automatic and non-automatic infusion techniques. A double blind trial of prostaglandin E2 and oxytocin given by intravenous infusion after amniotomy for induction of labour in 100 primigravidae with unfavourable induction features is reported. No clear-cut advantage of either drug emerged although PGE2 was perhaps superior when the cervix was highly unfavourable. Prostaglandin E2 appeared to produce less deleterious effects on the fetus but was associated with a higher incidence of maternal side effects. The automatic Cardiff Infusion apparatus was found to be a safe means of PGE2 infusion and to have advantages over the use of non-automatic techniques both for PGE2 and for oxytocin infusion.


The distribution within the placenta, myometrium, and decidua of 24Na-labelled hypertonic saline solution following intra-amniotic or extra-amniotic injection. 24Na-labelled hypertonic saline solution was injected into the uterus by the intra-amniotic or the extra-amniotic route, and six hours later determinations were made of the 24Na content of the placenta, myometrium, and decidua. The tissue specimens were obtained by subtotal hysterectomy. The activity of 24Na proved to be high in the decidua and in the fetal part of the placenta, but low in the myometrium. During labour, large amounts of prostaglandins have been found in the decidua, but little or none in the placenta. The present findings therefore lend further support to the assumption that hypertonic saline solution provokes abortion by inducing release of lysosomal phospholipase A2 into the cytoplasm of damaged decidual cells, resulting in prostaglandin synthesis and liberation.


A study of the effect of bromocriptine on serum oestradiol, prolactin, and follicle stimulating hormone levels in puerperal women. Nine normal women were examined in the first 11 days of the puerperium. Serum prolactin, oestradiol and follicle simulating hormone (FSH) levels were measured and the FSH response to 100 mug of intravenously administered follicle stimulating hormone/lutenizing hormone-releasing hormone (LH/FSH-RH) was assessed on day 1 post partum in five subjects and on days 4 (two subjects), 6 and 7 post partum in the other four subjects. Bromocriptine therapy was given to six of these women and four to seven days after the start of such therapy, when prolactin levels had fallen to normal non-pregnant levels, these women were retested. Before bromocriptine treatment, the basal FSH levels were unmeasurable in six and low in one other of the nine subjects. After bromocriptine treatment, the basal FSH levels were measurable in four out of six patients and were higher than in the untreated patients (P less than 0-01). The FSH response to LH/FSH-RH was unmeasurable in eight out of nine before treatment, which is less than the response seen in 26 normal men and women (P less than 0-01). There was also no discernible FSH response to LH/FSH-RH after treatment with bromocriptine. Before treatment with bromocriptine, the serum prolactin levels were elevated in all nine women but were within the normal range for non-pregnant women after three days of bromocriptine therapy. Thus postpartum gonadotrophin suppression seemed to depend on high prolactin levels and at least part of the antigonadotrophic action of prolactin seemed to be at pituitary level.


Effect of bromocriptine and chlorotrianisene on inhibition of lactation and serum prolactin. A comparative double-blind study. A double-blind study to compare the effect of 2-Br-alpha-ergocryptine (bromocriptine, CB 154, Sandoz) an ergot alkaloid with chlorotrianisene (Tace, Mer-National) on the inhibition of postpartum lactation was carried out in 38 women. At the dosages selected bromocriptine was more effective than chlorotrianisene in inhibiting lactation. Furthermore, bromocriptine significantly reduced serum prolactin levels to low normal values by the seventh day of treatment, whereas chlorotrianisene did not alter the normal progressive reduction in prolactin post-partum. Neither drug had significant toxic effects.


Structure of the modified nucleoside Q isolated from Escherichia coli transfer ribonucleic acid. 7-(4,5-cis-Dihydroxy-1-cyclopenten-3-ylaminomethyl)-7-deazaguanosine. The structure of the unknown modified nucleoside Q, which is present in the first position of the anticodons of Escherichia coli tRNA Tyr, tRNA His, tRNA Asn, tRNA Asp, is proposed to be 7-(4,5-cis-dihydroxy-1-cyclopenten-3-ylaminomethyl)-7-deazaguanosine (1). The structure of Q was deduced by means of its uv absorption, mass spectrometry, proton magnetic resonance spectroscopy, and studies of its chemical reactivity. The structure of Q is unique since it is a derivative of 7-deazaguanosine having cyclopentenediol in the side chain at the C-7 position. This is the first example of purine skeleton modification in a nucleoside from tRNA.


Sites of biological methylation of proteins in cultured chick muscle cells. The methylation of myosin and other proteins has been studied using primary cultures of 12-day-old embryonic chick leg muscle. The methyl group of [Me-3H] methionine is incorporated into basic amino acid residues with the formation of Nepsilon-monomethyllysine, Nepsilon-dimethyllysine, Nepsilon-trimethyllysine, 3-methylhistidine, NG-monomethylarginine, and NG-dimethylarginine which are isolated from acid hydrolysates of purified myosin, and of proteins from polysomes and from the cytosol of the cultured muscle cells. In the presence of 0.1 mM cycloheximide, incorporation of [Me-3H] methionine into the polysome-bound proteins was decreased to 16.3% of control levels with no change in the pattern of incorporation into the basic amino acid residues, although protein synthesis was inhibited 97.5%. When protein synthesis was allowed to resume in such cultures by the removal of cycloheximide, polypeptides containing labeled N-methylated residues were released from polysomes into the soluble fraction. Polypeptides containing N-methylated amino acids were also released from polysomes following treatment with 2 mM puromycin. Peptidyl-tRNA, isolated from ribosomes after exposure of cultures to [Me-3H] methionine, contained labeled N-methylated amino acids. When [Me-3H] methionine was incorporated in the presence of cycloheximide, the isolated peptidyl-tRNA still contained N-methylated amino acids although the amount of methylation was 22.4% of control levels. These experiments demonstrate that N-methylation of basic amino acid residues in proteins may occur while the polypeptide is still being synthesized on the ribosome. In addition, N-methylation can occur on the nascent polypeptides in the absence of protein synthesis.


Molecular interactions between ribosomal proteins. Evidence for specificity of interaction between isolated proteins. The proteins S2, S3, S5, and S10 from the 30S ribosomal subunit of Escherichia coli was studied by analytical ultracentrifugation to characterize them in solution and to determine whether isolated protein-protein interactions exist. Such interactions, if specific, may therefore bear some relationship to the spatial organization of the subunit structure. It was found that protein S2 self-associates to a slight extent and that solution mixtures of S2 and S3 contain only enough dimeric species to account for the S2 dimer. Hence, no observable interaction was detected between S2 and S3. Solution mixtures of proteins S5 and S10 revealed a species of molecular weight greater than either protein. The proposal is that S5 and S10 interact with an association equilibrium constant of 7.6 X 10(-5) M-1 at 3 degrees in a Tris buffer at pH 7.4. It was also shown that solution with a 1:1:1 mixture by mass, of S2, S5, and S10 contained a species possessing a molecular weight consistent with a simple ternary complex of the three proteins.


An immunological approach to the role of the low molecular weight subunits in myosin. II. Interaction of myosin and its subfragments with antibodies to the light chains. Immunological methods, in parellel with measurement of ATPase activity, have been used to characterize the reactions of antibodies specific for light chains with myosin and its water-soluble proteolytic subfragments, heavy meromyosin (HMM) and subfragment 1 (HMM S-1). Antiserum to the 5,5'-dithiobis(2-nitro-enzoic acid) (DTNB) light chain undergoes a precipitation reaction with all of the enzyme species, in which half of the homologous light chain is selectively dissociated. The results suggest that the incomplete dissociation reflects the way in which the light chain is bound, rather than the existence of two distinct species of DTNB 1.c. Little reaction was observed with antisera to alkali-released light chains, indicating that these components in myosin and the subfragments are either largely buried or else conformationally different from the isolated light chains used as immunogens. None of the antisera produced significant changes in Ca2+- or EDTA-ATPase activities. Moreover, calcium regulation through the troponin-tropomyosin system was unaffected by removal of DTNB 1.c. from myosin, as well as from the subfragments. The absolute level of actin-activated ATPase activity was, however, consistently lower in the presence of light chain antisera (or purified IgG and antibody) than in aqueous buffer or nonimmune serum. For both alkali and DTNB 1.c. antisera, this loss in activity seemed to result from steric hindrance of actin binding by antibody bound to undissociated light chain. Experimental conditions which would be expected to weaken such an antigen-antibody interaction, as well as the use of monovalent Fab in place of IgG, decreased the inhibition of activity. Altogether the activity measurements suggest that the light chains, particularly DTNB 1.c., are probably not integral parts of either the hydrolytic or actin-binding sites.


Evidence for photoinduced cross-linkage, in situ, of 30S ribosomal proteins to 16S rRNA. The effects of ultraviolet radiation on the 30S ribosomal subunit of Escherichia coli were studied. Irradiation in aqueous solution under anaerobic conditions resulted in a dose-dependent decrease in the separability of the rRNA and protein components of the 30S ribosomal subunit in 4 M urea-3 M LiC1. The results of gel filtration studies of the irradiated ribosomes before and after treatment with pancreatic ribonuclease indicated that the decrease in separability of the ribosome components was a result of the photoinduced formation of covalent RNA-protein cross-links. The number of covalent cross-links was estimated to correspond to less than 3 per 10,000 daltons of ribosomal proteins. One-dimensional gel electrophoresis studies of the course of the photoinduced cross-linkage reaction indicated that cross-linkage of individual 30S ribosomal proteins to the 16S rRNA proceeds in two dose-dependent steps. The first step requires an input of 1 X 10(20) quanta of 253.7 -nm radiation and results in the cross-linkage of at least five ribosomal proteins to the 16S rRNA. The second step requires a total input of 2 X 10(20) quanta of 253.7 -nm radiation, and results in the cross-linkage of most of the remaining 30S ribosomal proteins to the 16S rRNA.


A procedure for the rapid, large-scall purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precipitation and DNA-cellulose chromatography. An improved method is described for the purification of the DNA-dependent RNA polymerase [ribonucleosidetriphosphate:RNA  nucleotidyltransferase,  EC 2.7.7.6] from Escherichia coli. The method involves lysozyme-sodium deoxycholate lysis, low-speed centrifugation, precipitation with Polymin P, elution from the Polymin P precipitate, ammonium sulfate precipitation, and chromatography on DNA-cellulose and Bio-Gel A 5m. RNA polymerase is purified to electrophoretic homogeneity in 2 days with a recovery of 45%, resulting in a yield of 250 mg of holoenzyme from 500 g of cells.


A fluorometric study of DNA-bound benzo[a]pyrene. Comparisons were made among the fluorescence spectra of DNA-bound benzo[a]pyrenes which were produced in vivo and in vitro. DNA from mouse skin treated with benzo[a]pyrene had a maximum emission beyond 400 nm, which was clearly distinguished from that of DNA-bound benzo[a]pyrene 4,5-oxide. The emission spectra from mouse skin were classified into two groups, type I and type II. The former was similar to the spectrum of benzo[a]pyrene, although the two maxima were shifted to longer wavelengths (410 and 435 nm). Type II was characterized by a broad peak around 430 nm. Type I and type II were obtained from different fractions of hydroxylapatite chromatography, but type I was changed into type II during storage. This suggests that type II is a modified product of type I. The emission spectra of both groups also were detected in in vitro activating systems, including photoirradiation, iodine treatment, and hydrogen peroxide treatment. Treatment of Escherichia coli with benzo[a]pyrene during culture produced only fluorescence of type I. Although the relationship between types I and II remains to be established, both types of fluorescence evidently indicate that the conjugated ring structure of the parent compound, benzo[a]pyrene, is preserved intact in DNA-bound benzo[a]pyrene. Several lines of evidence suggest that the proximate (active) form is an unidentified hydroxylated product including an oxy radical, but a cation radical cannot be completely excluded.


On the mode of action of lomofungin, an inhibitor of RNA synthesis in yeast. Lomofungin is a potent inhibitor of RNA synthesis in yeast. Studies on the mode of action of the inhibitor were carried out using yeast RNA polymerases A and B and bacterial RNA polymerase. In vitro inhibition of RNA synthesis is independent of the nature and concentration of the template used and of the nucleoside triphosphate concentration. The extent of inhibition is strongly dependent upon the nature and concentration of divalent cations used to simulate transcription. The three RNA polymerases were inhibited to the same extent in the presence of Mn2+ ions whereas little inhibition was observed with Mg2+ ions. Spectrophotometric studies reveal the formation of different complexes between lomofungin and divalent cations (Mn2+, Mg2+, or Zn2+) with the respective stoichiometries of 0.5, 1, and 2 divalent cations per molecule of lomofungin. The complexes formed depend upon the nature of the divalent cation involved. No direct interaction between lomofungin and DNA could be observed in the presence of divalent cations but evidence is presented that lomofungin interacts with yeast RNA polymerase A. Inhibition of RNA synthesis occurs at the level of both chain initiation and elongation.


T7 RNA polymerase: conformation, functional groups, and promotor binding. Circular dichroic spectra of T7 RNA polymerase show minima at 222 nm ([theta]m=-7.9 X 10(3) deg cm2/dmol) and 208 nm ([theta]m =-7.55 X 10(3) deg cm2/dmol) and a maximum at 193 nm ([theta]m = 1.2 X 10(4) deg cm2/dmol). The small mean residue ellipticity above 200 nm indicates that the secondary structure contains approximately 12% alpha helix. The secondary structure is unaltered by high salt, glycerol, -SH reagents, nitration of tyrosyl residues, and chelating agents. Binding of the native enzyme to [32P]T7 DNA has been measured by the retention of the protein-[32P]DNA complexes on nitrocellulose filters. At 37degrees T7 RNA polymerase binds to its promoters in the absence of NTP's. Binding and catalytic activity are both abolished at 0degree. Binding of the initiating [gamma-32P]GTP can also be detected by the filter binding assay. Native T7 RNA polymerase is inactivated by reaction with 1 mol of 5,5'-dithiobis(2-nitrobenzoic acid) (Nbs2) or 1 mol of [14C]iodoacetamide. The latter reaction is blocked by Nbs2 suggesting that a single -SH group is required for activity. Alkylation of the -SH group does not alter binding of the enzyme to the DNA template, but modifies the binding of GTP to the enzyme. Nitration of approximately4 surface tyrosyl residues of the protein prevents binding to T7 DNA. The restriction endonuclease, Hpa II, cuts T7 DNA into approximately40 fragments and reduces total RNA synthesis by T7 RNA polymerase by 70%. Fragmentation of the DNA template by Hpa II does not alter the rate of RNA chain initiation by T7 polymerase, and restriction fragments accounting for approximately25% of the T7 DNA still bind tightly to the enzyme. Thus the T7 RNA polymerase promoters remain intact on the restriction fragments. Gel electrophoresis of the transcription products, using restriction fragments as templates, show that of the seven in vitro transcripts produced by T7 RNA polymerase from whole T7 DNA, only the smallest (representing the last 1.5% of the genome) is transcribed from Hpa II fragments. The remaining transcripts are replaced by six new and much shorter mRNA's. The DNA fragments containing the promoters for these mRNA's have been removed from the fragment mix by binding them to the enzyme and retaining the complexes on nitrocellulose filters.


Transcription of fractionated calf thymus chromatin by RNA polymerase of calf thymus and Escherichia coli. Calf thymus chromatin has been sheared and fractionated on sucrose gradients. Approximately 5-10% of the chromatin is resolved from the bulk of the imput chromatin as a slowly sedimenting fraction. The protein/DNA ratio of the slowly sedimenting fraction is not greatly different from the protein/DNA ratio of the more rapidly sedimenting chromatin fraction. Analysis of DNA of the chromatin fractions by CsC1 equilibrium density gradient centrifugation indicates that DNA of the slowly sedimenting fraction is depleted in the satellite DNA banding at 1.716 g/cm3. The template properties of the chromatin fractions have been examined with Escherichia coli RNA polymerase and form II and form III RNA polymerases of calf thymus. At rate-limiting concentrations, the slowly sedimenting fraction is twofold more active than the rapidly sedimenting fraction as a template for E. coli RNA polymerase. Homologous form II and form III RNA polymerases are respectively 30-fold and 16-fold more active with the slowly sedimenting fraction than the rapidly sedimenting fraction. The activity of form II RNA polymerase in transcribing the slowly sedimenting fraction exceeds its activity in transcribing an equal concentration of native DNA. Kinetic studies, in which RNA polymerase activity is assayed at various concentrations of chromatin, indicate that the greater activity of E. coli RNA polymerase with the slowly sedimenting fraction is due to an increased rate of transcription at saturating concentrations of template (Vmax), and is not due to a lower concentration required for half-maximal rate of transcription (Km). In contrast, the increased rates of transcription of the slowly sedimenting chromatin faction by the homologous polymerases are due to a decrease in concentration required for half-maximal rate of transcription rather than an increased rate of transscription at saturation concentrations of template. The relative degrease of satellite DNA in the slowly sedimenting fraction of chromatin and the enhanced template activity of the slowly sedimenting fraction suggest that this fraction is equivalent to nuclear euchromatin while the more rapidly sedimenting chromatin is equivalent to nuclear heterochromatin.


L-phenylalanine:tRNA ligase of Escherichia coli K10. A rapid kinetic investigation of the catalytic reaction. The kinetics of the amino acid activation and the transfer of the amino acid to tRNA have been investigated for L-phenylalanine:tRNA ligase of Escherichia coli K10 by stopped-flow and radioactive techniques. The rapid kinetics were followed by the observation of the displacement of the fluorescent dye, 6-p-toluidinylnaphthalene-2-sulfonate from the binding site of L-phenylalanine under conditions where a single active site of the enzyme was involved. The following results are of particular interest. (1) Equilibrium binding of L-phenylalanine and tRNAPhe indicates in each case two sites of interaction with an approximately tenfold difference of the binding affinity. (2) Experimental conditions of the kinetic investigation were chosen to favor reactions at the high affinity binding sites. Under those conditions, the rate constants have been evaluated at 1 mM magnesium to be in the range 12-25 sec-1 for the activation reaction and 42-77 sec-1 for the reverse, the variation of the values depending on those of the dissociation constants used for computation. The rate constant for the transfer reaction is 0.05 sec-1 and for the reverse 0.19 sec-1. The forward reaction is rate limiting for the overall reaction at single turnover and steady-state conditions. (3) All rate constants depend on the concentration of magnesium. Evidence is provided that the transfer occurs via a productive enzyme-tRNAPhe complex which is in a magnesium-dependent equilibrium with an unproductive complex, high magnesium favoring the former. The position of the tRNA-CCA end in the productive complex is such, that the fluorescent dye can be displaced by Phe-tRNAPhe. The thermodynamics of the overall reaction have been treated on the basis of the partial reactions. The free enthalpy of the completed reaction was calculated to be very close to zero. The significance of the adenylate intermediate is discussed with respect to the product inhibition expected on the basis of the tendency of tRNAPhe and L-phenylalanine to form tight complexes with the enzyme.


Polynucleotide analogs: acrylic acid and maleic acid copolymers of 1-vinyluracil and 9-vinyladenine. Radical-induced copolymerization of 1-vinyluracil and maleic anhydride gave, after hydrolysis, a polymer containing a 1:1 monomer ratio of 1-vinyluracil-maleic acid. Gamma-Ray-induced copolymerization of 1-vinyluracil with acrylic acid gave a polymer with a ratio of 1:1.7. Similar treatment of 9-vinlyadenine and acrylic acid resulted in a polymer with a 1:3.2 ratio. These three compounds are potent stimulants of poly (uridylic acid) coded polyphenylalanine synthesis in an in vitro cell free system purified from Escherichia coli MRE 600. The double-stranded polymer, poly(inosinic acid)-poly(cytidylic acid), also stimulates polyphenylalanine synthesis in this assay.


Inactivation of rat liver RNA polymerases I and II and yeast RNA polymerase I by pyrodixal 5'-phosphate. Evidence for the participation of lysyl residues at the active site. Purified DNA-dependent  RNA polymerase forms I (A) and II (B) from rat liver and form I from yeast are rapidly inactivated by pyridoxal 5'-phosphate at pH 8.0. The inhibition is relatively specific since pyridoxamine 5'-phosphate is not an inhibitor and pyridoxal is about 12 times less effective than pyridoxal 5'-phosphate. The inactivation is reversed by high concentrations of amines, and can be made irreversible by reduction with NaBH4. Spectral analysis of the inhibited enzyme and its NaBH4 reduction product indicates that a Schiff base forms between the aldehyde group of pyridoxal 5'-phosphate and one or more amino groups of the protein. Nepsilon-Pyridoxyllysine was identified as the only product in acid hydrolysates of the reduced yeast RNA polymerase I-pyridoxal 5'-phosphate complex. Complete inactivation of yeast polymerase I results in the incorporation of 3-4 mol of pyridoxal 5'-phosphate/1 mol of enzyme. DNA and nucleotide substrates partially protect the enzymes from inactivation. These results suggest that one or more lysyl amino groups are critical for the activity of animal RNA polymerases and show that pyridoxal 5'-phosphate is a suitable probe for studying the active sites of these enzymes. Comparison of the present results with those previously obtained with Eschericha coli RNA polymerase in this laboratory suggest a new degree of structural homology between eucaryotic and procaryotic RNA polymerases.


The effect of salt on phospholipid fatty acid composition in Escherichia coli K-12. The fatty acid compositions of the three major phospholipids of Escherichia coli K-12; phosphatidylethanolamine, phosphatidylglycerol and cardiolipin; were determined during growth in media differing in NaCl concentration. Significant differences in fatty acid composition of the phospholipids were observed in the stationary phase cultures, but no appreciable differences were found in early exponential cultures.


Nitrogenases of Klebsiella pneumoniae and Azotobacter chroococum. Complex formation between the component proteins. 1. Sedimentation velocity analyses of mixtures of highly purified component proteins of Azotobacter chroococcum are consistent with the formation of a tight 1 : 1 complex in the absence of Na2 S2 O4. 1 : 1 complex formation between complementary proteins from A. chroococcum and Klebsiella pneumoniae was also observed. The addition of 5 mM Na2 S2 O4 weakened the interaction between the A. chroococcum proteins and also the interaction between complementary proteins of A. chroococcum and K. pneumoniae. 2. Steady-state kinetic data for acetylene reduction at low protein concentrations have been used to calculate association constants at 30 degrees C for the 1 : 1 protein complexes of nitrogenase proteins from A. chroococcum, K. pneumoniae and mixtures of complementary proteins from both organisms. Values centered around 3 - 10(7) M-1 were obtained. 3. The temperature dependence of the association constant for the complex formed by the K. pneumoniae proteins exhibited a sharp break at 17 degrees C with deltaH = 0 and deltaH = 418 kJ - mol-1 above and below 17 degrees C, respectively. 4. The Arrhenius plot for acetylene reduction by the complex formed by the K. pneumoniae proteins was linear over the range 12-40 degrees C with deltaH = 80 kJ - mol-1.


Effect of L-azetidine 2-carboxylic acid on growth and proline metabolism in Escherichia coli. The effects of L-azetidine 2-carboxylic acid on growth and proline metabolism in a proline-requiring auxotroph of Escherichia coli are described. The homologue inhibited growth of the wild type and it, alone, did not substitute effectively for proline as a growth supplement for the mutant. In medium containing 0.05 mM proline, the addition of increasing amounts of homologue progressively inhibited growth of the wild type but stimulated growth of the mutant at homologue: proline ratios of 10 : 1 and 50 : 1. This suggested that the homologue exerted a "sparing effect" on proline in the mutant. The incorporation of L-[U-14C]proline and L-[3H]azetidine 2-carboxylic acid into hot trichloroacetic acid-insoluble material in the mutant was measured. Amino acid analysis of the insoluble material from cells incubated with radiolabeled proline alone revealed that proline was partially degraded and metabolized to other amino acids prior to incorporation into protein. The addition of unlabeled homologue to the incubation medium significantly reduced proline catabolism, suggesting that the homologue exerted a sparing effect on proline in this mutant. In medium containing unlabeled proline and radiolabeled L-azetidine 2-carboxylic acid, the homologue was incorporated both intact and partially degraded prior to incorporation into protein. Alanine was the major L-azetidine 2-carboxylic acid catabolite.


External yeast beta-fructosidase. The role of tryptophyl residues in catalysis. 1. In native invertase at pH 4.6, 23 out of a total of 34 tryptophyl residues are "exposed" to oxidation with N-bromosuccinimide, the other residues being apparently shielded from the oxidant within the molecule. 2. Oxidation of 5-6 tryptophyl residues/molecule with N-bromosuccinimide is proportional to the complete inactivation of the enzyme, and appears to be specific for indole chromophore only. The ligand binding and fluorescence measurements indicate that the oxidation of native enzyme, up to 50% inhibition, apparently does not change the conformation and topography of enzymes surface. 3. Invertase is inhibited by diazonium-1-H-tetrazole. Since tyrosine residues can be excluded by nitration studies as catalytically unimportant, it appears that a mocification of a single histidyl residue/molecule with diazonium-1-H-tetrazole is sufficient to abolish the enzymic activity. However, the absence of inhibition with diethyl pyrocarbonate indicates that the inhibition with diazonium-1-H-tetrazole may be mediated through steric hindrance or other indirect effects. 4. The absence of inhibition with 2,4-dinitrophenylhydrazine, trinitro benzenesulfonic acid and 5,5'-dithiobis-(2-nitrobenzoate) indicates that the carbonyl groups of the carbohydrate moiety, free amino and -SH groups are not essential for activity.


Pyruvate decarboxylase III. Specificity restrictions for thiamine pyrophosphate in the protein association step, sub-unit structure. Pyruvate decarboxylase dissociates into sub-units of one half the molecular weight at alkaline pH. At the same conditions the cofactors thiamine pyrophosphate and Mg2+ are released and can be separated from the protein. Thiamine pyrophosphate is an obligatory cofactor for reconstitution to the oligomer [1]. In this study the effect of thiamine pyrophosphate derivatives (thiamine monophosphate, thiamine, and thiazole pyrophosphate) upon the reconstitution procedure was evaluated. The complete association of sub-units to form active oligomer was attained only when thiamine pyrophosphate was present. It is concluded that both the pyrimidine ring and the pyrophosphate group are required for productive co-enzyme binding and it is proposed that this interaction effects a conformational change which promotes protomer aggregation to form the enzymatically active holoenzyme. In addition data are presented which indicate that the monomer unit is 60 000 +/- 3000 daltons and that the N-terminal amino acid is histidine. Since the molecular weight of the active oligomer is 230 000 it is proposed that pyruvate decarboxylase is a tetramer comprised of four identical or nearly identical monomer units.


Multiple forms of mitochondrial alcohol dehydrogenase in Saccharomyces cerevisiae. Three forms of NAD-dependent alcohol dehydrogenase have been characterized in yeast by their heat sensitivity, their specificity and their electrophoretic patterns. Thermal stability increases in the following order: alcohol dehydrogenase I (fermentative enzyme), alcohol dehydrogenase II (oxidative enzyme), alcohol dehydrogenase III (mitochondrial enzyme). Work with isolated mitochondria shows that alcohol dehydrogenase III is the only form of alcohol dehydrogenase present in these organelles. Starch gel electrophoresis of alcohol dehydrogenase III reveals an active zone of slow migration which consists of five sub-bands. The relative activity of these five sub-bands varies with the conditions of growth. Mitochondrial alcohol dehydrogenase represents never more than 10% of the total cellular alcohol dehydrogenase. Information for its biosynthesis seems to be located in nucleic DNA. The mitochondrial enzyme shows a high affinity for alcohols with a double bond conjugated to the alcohol function.


Synthesis and degradation of early mRNA in lambda phage. Using two different Escherichia coli mutants defective in elongation factors EFG and EFTs required for peptide synthesis, lambda phage with or without a tof mutation was analysed for synthesis of early mRNA by DNA-RNA hybridization technique. (1) In CP78G carrying temperature-sensitive elongation factor G, shift-up to high temperature (41 degrees C) in the middle of phage infection did not affect early mRNA synthesis with lambdatof+ phage but did inhibit it with lambdatof- phage. (2) In HAK88 carrying temperature-sensitive elongation factor Ts, shift-up to 41 degrees C dropped total cellular RNA synthesis to below 10-20 percent of the control in the presence or absence of phage infection (stringent control). Under such a condition for suppressed protein synthesis, lambda early mRNA synthesis was completely blocked with both tof+ and tof- phage infection. Thus, stringent control seems to exert its effect on lambda early mRNA synthesis. (3) Addition of chloramphenicol, which is known to relax the stringent control of RNA synthesis, to the culture of phage-infected HAK88 at 41 degrees C resulted in full recovery of tof+-mRNA synthesis, but only in partial recovery to tof --mRNA synthesis. (4) Analysis of the stability of lambda mRNA indicated an exponential decay, and a halflife of tof --mRNA of 6 min when that of tof+-mRNA was 13 min at 37 degrees C.


Aminoacylation of Phaseolus vulgaris cytoplasmic, chloroplastic and mitochondrial tRNAsMet and of Escherichia coli tRNAsMet by homologous and heterologous enzymes. Met-tRNA synthetase from Paseolus vulgaris cytoplasm could be separated from its chloroplastic or mitochondrial counterpart by DEAE-cellulose chromatography, but the Met-tRNA synthetase from the two latter organelles could not be distinguished using DEAE-cellulose, hydroxyapatite or CM-Sephadex chromatography. As revealed by reverse-phase chromatography, bean cytoplasm contains 2 tRNAsMet; only one is charged by chloroplast, mitochondrial or Escherichia coli Met-tRNA synthetase. Mitochondria contain, in addition to the 2 cytoplasmic tRNAsMet, 3 mitochondria-spedific tRNAsMet; 2 can be formylated by the mitochondrial or the E. coli transformylase; all 3 are charged by mitochondrial, chloroplastic or E, coli Met-tRNA synthetase; none is charged by the cytoplasmic enzyme. Chloroplasts contain, in addition to the 2 cytoplasmic tRNAsMet, 3 chloroplast-specific tRNAsMet, different from the mitochondrial tRNAsMet; one is formylatable by the chloroplastic or the E. coli transformylase; all 3 are charged by chloroplastic, mitochondrial or E. coli Met-tRNA synthetase; only one is charged by the cytoplasmic enzyme. Of the 3 E. coli tRNAsMet, only the formylatable species can be charged by bean cytoplasmic, chloroplastic or mitochondrial Met-tRNA synthetase.


Substrate specificity of Salmonella typhimurium RNAase III and the nature of products formed. RNAase III, an endonuclease specific for double-stranded substrates, has been obtained in a highly purified form from extracts of Salmonella typhimurium. Poly (I-C) and a mixture of poly(A) and poly(U) (1 : 1), the latter in presence of 5 mM Mg2+, act as excellent substrates. Poly(I-C) is only partially hydrolysed by RNAase III and the product or products are acted upon by both RNAase I and RNAase II indicating that the products may be oligonucleotides. This conclusion is supported by two-dimensional chromatography on DEAE paper.


[Study of the degradation reaction of paranitrosodimethylaniline by a C-nitrosoreductase of yeast]. We have purified, by a three step procedure, a yeast C-nitrosoreductase and studied its action on p-nitroso-N,N-dimethylaniline. Microcalorimetric measurements performed on the microcalorimeter LKB "batch", have been given the number of protons exchanged during the reaction : four protons are involved. Two are given by NADH and the others are supplied by the buffer. We could observe two steps in the catalytic mechanism. The enthalpy change of the first one is greater than that of the second. Spectrophotometric and polarographic experiments corroborate this results : two enzymatic steps could be demonstrated. The reduction of p-nitroso-N,N-dimethylaniline is characterized by a peak with a half wave potential at -0,31 V (in M Tris-HCl, pH 8,30). While the amplitude of this one decreases, a second wave appears at -1,03 V.


[Analysis of a small quatity of polypeptides by two-dimensional electrophoresis: application to characterization of proteins]. A new two-dimensional electrophoresis is described. The first electrophoresis is performed in a 10 p. cent gel (1 X 110 mm) in the presence of 7 M urea; the migration in the second dimension proceeds on a 20 p. cent polyacrylamide sheet (160 X 110 X 1 mm) in the presence of sodium dodecyl sulphate. This system resolves most of the polypeptides obtained by treating a very small amount (about 100 mug) of proteins with cyanogene bromide. Macromolecules can thus be quickly and easily characterized.


[Quaternary structure of Escherichia coli polynucleotide phosphorylase: chemical characterization of form A]. We had previously shown the existence of two classes of polynucleotide phosphorylases : Form A which is made of alpha chains and carries the catalytic sites ; and form B which is constituted of alpha chains and of beta chains. We performed some chemical analyses of form A (N-terminal sequence, amino acid composition, peptide mapping) which suggest that the alpha chains are all identical and moreover that they have no relationship with the beta chains. The latter do not therefore derive from a partial proteolytic degradation of the alpha chains and can therefore be considered as true subunits of the enzyme.


Effects of changes in growth rate on the levels of several aminoacyl-tRNA synthetases in yeast. When the growth rate of yeast cells is decreased (for instance by transferring the cells from a rich medium to a poor one, or when the cells enter the stationary phase, or when growth is inhibited by cycloheximide, or when valine is removed from the medium supporting growth of a valine-requiring mutant, or when a thermosensitive mutant is shifted to the non-permissive temperature) there is a decrease in the levels of the four aminoacyl-tRNA synthetases tested. Conversely, an increase in the growth rate is accompanied by an increase in the levels of the four enzymes. But when the growth rate is slowed down by decreasing the temperature of the medium, no effect on the levels of the aminoacyl-tRNA synthetases is observed. These results are consistent with the concept of "metabolic regulation" proposed by Parker and neidhardt.


Nuclear power for the artificial heart.

Nuclear power for the artificial heart. Studies showed that a totally implantable nuclear-powered artificial heart is practicable, and consequently a prototype system development program was initiated in July 1973. Basic testing of the prototype should be completed in 1977, with extensive studies to qualify the device for clinical use running at least into the early 1980s.


Neurophysiological and behavioral organization of premature infants in the first months of life. Many explanations for the current findings of limited group differences between premature and term infants at comparable conceptional ages can be imagined that remain buried in the data as currently presented. For example, some prematures may indeed be advanced in many areas of behavioral and neurophysiological development, but this is counterbalanced by prematures who are significantly delayed in these parameters for medical reasons such as hypoxia or metabolic disturbance, while a third portion of the infants has uneven and disorganizing mixtures of delayed and advanced development in selective modalities. Hopefully future research which takes these various possibilities into consideration will give us a better understanding of this complex problem.


Some thoughts on experimental screening. Experimental screening of anticancer drugs is discussed from the standpoint of the need for close co-operation between experimentalists and clinicans in developing therapies for human trials. Particular emphasis is given to screening analogues of known active drugs including the approaches in chemical synthesis and tests of experimental tumor activity, as well as the questions that determine clinical interest and capabilities for human trials. In the area of combinations of two or more drugs, the need for experimental systems providing data for rational design of clinical regimens is highlighted. Means for experimental evaluation of the major variables of dose ratio, dose schedule and sequence of drug administration are considered. Finally, the overall problems of drug screening are discussed within the context of combining drugs with other treatment modalities, such as surgery, radiotherapy, and immunotherapy. The complexities of designing better cancer treatment will require close interaction between experimental and clinical studies.


Cellulase and protein production from mixed cultures of Trichoderma viride and a yeast. Fermentations with mixed cultures of the cellulolytic fungus Trichoderma viride and the yeast Saccharomyces cerevisiae or Candida utilis were examined. The fermentations were carried out in an aerated 5 liter fermentor with NaOH treated barley straw as the cellulose source (2-4%). Yeast was inoculated 24-32 hr after the fungus and the growth of the two organisms was followed through the production of CO2 and cell protein. In comparison with fermentations with T. viride alone, the production time for maximum yields of cellulases and cell protein was reduced by several days, depending on the straw concentrations. The protein content of the growth product was 21-22% and the amino acid composition of the product resembled that of T. viride alone.


[Effect of embichin (HN2) and its monofunctional analog (HN1) on chromatin and DNA matrix activity in an in vitro RNA-polymerase system]. Embichin inhibited the matrix activity of chromatin and DNA in the RNA-polymerase system in vitro much more than its monofunctional analogue. Chromatin possessed a greater sensitivity to the action of embichin in comparison with the deproteinised DNA. However,  with the action of a monofunctional embichin analogue there was a greater reduction of the matrix activity of DNA in comparison with chromatin. The depression mechanism of the matrix activity of chromatin with the action of embichin was apparently associated with the capacity of the latter to form the DNA-protein bonds in the chromatin composition.


[Development and clinical approval of a micromodification of the human leukocyte migration inhibition reaction]. The author suggests a new modification of the human blood leukocyte migration inhibition test. The results of the clinical approval of the test permit it to be recommended for the investigation in dynamics of the cellular immunity and the humoral factors in ptients with different immunopathological state. This modification permits to carry out 10-15 parallel investigations using small quantitites of the blood.


[Study of the enzymatic activity of the blood leukocytes in experimental nephrotoxic nephritis in rats]. The enzymatic activity of blood leukocytes was studied in rats with Masugi nephritis by means of the quantitative cytochemical method. Nephrotoxin injection during first hours and days did not produce specific changes in the enzymatic activity. Maximum changes were observed on the 8-th day when the elevation of the activity of dehydrogenase lymphocytes and neutrophils acid phosphatase was observed at the same time with most pronounced histological changes in the kidneys, the decrease of the complement titer and the presence of nephrotoxin and autoglobulin in the renal glomeruli. Later the histological changes became less pronounced, and the enzymatic activity decreased. In rats with proteinuria the activity of the neutrophil leukocytes acid phosphatase was higher than in animals without proteinuria.


[Use of an immunothermistographic method for detecting humoral antibodies in the allotransplantation of skin and cornea in rabbits]. Humoral antibodies were investigated by the method of immunothermistography in 43 rabbits after allotransplantation of the skin and cornea. The method of immunothermistography is based on the changes in thermoconduction of the medium as a result of the antigen-antibody reaction. The antibodies were discovered in the blood serum 3 to 5 days after the skin transplantation. The antibody titre gradually increased. Retransplantation of the skin at first decreased the concentration of antibodies, but the antibody titre rose in 8-12 days. In most of the animals the antibodies disappeared in 2-3 months. The same data were obtained in transplantation of the cornea. A correlation between thermistography data and Hougher's reaction and also the dynamics of the complement titre was found. The method of immunothermistography aids in discovery of the humoral tissue antibodies at the early period of the rejection reaction; it is highly sensitive and simple.


[Organ culture of human embryonic lung tissue]. Results of morphological study of organotypical cultures of the embryonal pulmonary tissue of 22-24 weeks human fetuses showed that for 30 experimental days they passed the period of adaptation, of the optimal growth and differentiation and gradual death. These periods were also characteristic of organic cultures of the embryonal pulmonary tissue of rodents. However, the processes of growth and differentiation in the corresponding human cultures were seen chiefly in the connective, and in rodents - in the epithelial tissue.


Potato avoidance during pregnancy in women with a previous infant with either anencephaly and/or spina bifida. This investigation is a direct attempt to test Renwick's (1972) hypothesis that 95% of anencephaly and spina bifida (ASB) is preventable by the avoidance of potatoes during pregnancy. Although the numbers involved in the study are small, the investigation fails to support the concept that short-term avoidance of potatoes before conception and throughout pregnancy in women who have had a previous ASB infant reduces the recurrence risk. In the potato-free group, of 23 pregnancies which went to term two infants had ASB (8-7%); whereas in the non-potato-free group, of 56 which went to term two infants had ASB (3-6%). The recurrence risk in both groups was about 5%. The incidence of ASB in the groups shows no significant difference (P = 0-58) and in the potato-free group was not reduced by 95% as postulated by Renwick.


Recent advances in acoustical microscopy. One cannot be involved in research in universities, government establishments and industry for 35 years without becoming aware of a number of misconceptions about pure science, applied science, engineering and technology in the mind of the public and also in the very places where research is carried out. There is confusion about the aims of research and development. There is uncertainty about the proper roles of universities, research establishments and industry. There is the question whether science is intrinsically good or not. The author illuminates some of these issues by relating milestones in the development of microscopy--optical as well as electron--and gives a snapshot picture of the recent work at Stanford University on the acoustic microscope as a felicitous instance of physics applied to the ever-present desire of mankind: to explore the unknown and to understand nature. Experiments with a mechanically scanning type of acoustic microscope have shown that a resolution limited only by diffraction can be attained and using a sonic frequency of 1-2 GHz, objects of less than 1 mum diameter can in principle be resolved. Various biological tissues have been explored and it appears that the acoustical images contain information substantially different from that obtained optically or electron-optically. Images have also been obtained using the harmonic radiation generated within the object and this mode of operation promises to open up yet another dimension of information. When, in due course, the full power, and the limitations, of the acoustic microscope are appreciated, its use may well lead to advances in science and technology comparable to those brought about by the introduction of the electron microscope.


The management of urinary fistulae following cadaveric renal transplantation. Vesicoureteric, ureteric and calyceal fistulae in cadaver transplants may be successfully treated by early exploration and nephrostomy splintage with silastic tubing for a period of not less than 6 weeks.


The microbial flora of the vagina and its relationship to bacteriuria in diabetic and non-diabetic women. The vaginal and urinary flora have been studied in 57 diabetic and 33 non-diabetic patients. In both populations, no correlation was found between urogenital symptoms and colonisation of the vagina. There was no greater prevalence of potentially pathogenic organisms in diabetic patients, even when control was poor. However, although not statistically significant, group B streptococci were isolated more frequently from poorly controlled diabetics. Bacteriuria was not significantly more common in diabetics. Gram-negative bacilli were found in the vagina significantly more often in bacteriuric patients; the prevalence of such organisms being four-fold greater after the menopause.


Antibiotic resistance and survival of E coli in the alimentary tract. Some antibiotics tend to select for R-factor-carrying Escherichia coli in the human gut, with complex long-term consequences. Some resistant strains disappear rapidly when treatment ends, while others persist for months in the absence of obvious antibiotic selection pressure, and the performance of individual resistant strains seems to depend more on the nature of the strain than on the plasmid carried. R plasmids are relatively rare in those E coli that colonize well in the gut and resistant bacteria therefore tend to disappear when treatment ends, but this situation could change dramatically if R plasmids became prevalent among those strains of E coli that colonize effectively.


The forgotten nodule: complications of sacral nodules in rheumatoid arthritis. Nodules commonly occur in rheumatoid arthritis and occasionally give rise to complications. The sacral nodule is easily missed and may ulcerate to produce extensive sacral sores which may lead to serious and even fatal complications in patients with rheumatoid arthritis. Seven cases are reported which illustrate some of these features.


Which elements are excited in electrical stimulation of mammalian central nervous system: a review. (1) There are data on the amount of current necessary to stimulate a myelinated fiber or cell body and/or its axon a given distance away from a monopolar electrode over the entire range of practical interest for intracranial stimulation. Data do not exist for other electrode configurations. (2) Currents from a monopolar cathode of more than 8 times threshold may block action potentials in axons. Therefore, only axons lying in a shell around the electrode are stimulated. Elements very close to the electrode may not be stimulated. Close to an electrode small diameter axons may be stimulated and larger ones may not be. (3) Most, and perhaps all, CNS myelinated fibers have chronaxies of 50-100 musec. When gray matter is stimulated, the chronaxie is often 200-700 musec. It is not clear what is being stimulated in this case. Current-duration relations should be determined for many more responses. (4) There are no current-distance or current-duration data for central finely myelinated or unmyelinated fibers. (5) It takes less cathodal current than anodal to stimulate a myelinated fiber passing by a monopolar electrode. When a monopolar electrode is near a cell body, on the opposite side from the axon, often the lowest threshold is anodal, but sometimes cathodal. Stimulation of a neuron near its cell body is not well understood, but in many cases the axon is probably stimulated. (6) Orientation of cell body and axons with respect to current flow is important. For an axon it is the component of the voltage gradient parallel to the fiber that is important. (7) The pia has a significant resistance and capacitance. Gray matter, white matter, and cerebrospinal fluid have different resistivities, which affect patterns of current flow. (8) More is known about stimulation of mammalian CNS than most workers are aware of. Much of what is unknown seems solvable with current methods.


Carboxyl-terminal sequences of procaryotic ribosomal proteins from Escherichia coli, Bacillus stearothermophilus, and Halobacterium cutirubrum. The carboxyl-terminal amino acid sequences of two ribosomal proteins, Escherichia coli L12 and E. coli S4, and the proteins believed to be their equivalents from Bacillus stearothermophilus and Halobacterium cutirubrum, were determined. These proteins are known to be required for peptide chain termination (L12) and in ribosome assembly (S4). The carboxyl-terminal sequences obtained suggest that the E. coli and B. stearothermophilus proteins have retained structural homology in this region, whereas the H. cutirubrum proteins have not.


Reconstitution of 48 S particles with subunit association activity from Escherichia coli 50 S ribosomal components. Previous studies have shown that 50 S ribosomal proteins bind sequentially to RNA's of the 50 S subunit via temperature-dependent steps (0 leads to 37 leads to 55 degrees C) to form 28, 32, (43) and 48 S particles (Chu, F.K., and Maeba, P.Y. (1973) Can. J. Biochem. 51, 129-139). When three equivalents of 50 S ribosomal proteins are used per equivalent of 23 S RNA, the yield of reconstituted 48 S particles is greatly increased. The addition of 10 mM spermidine increases the rate of reconstitution and abolishes the need for prior incubation at lower temperatures when 48 S particles are formed at 55 degrees C. In the presence of excess 50 S ribosomal proteins and 10 mM spermidine, 48 S particles formed at 50 degrees C are able to associate with 30 S subunits in a stoichiometric manner to form 70 S complexes. These 48 S particles compete effectively with 50 S subunits for 30 S subunits and have an association activity equal to that of 50 S subunits. As compared to the 34 proteins in 50 S subunits, 4 are either absent or present in marginal amounts, 11 in reduced amounts, and 19 in approximately normal amounts in the 48 S particles.


Nuclear demethylation and C-24 alkylation during ergosterol biosynthesis in Saccharomyces cerevisiae. The role of 4,4-dimethylzymosterol (3), 4,4-dimethylfecosterol (4) and 31-norlanosterol (5) in the biosynthesis of ergosterol in Saccharomyces cerevisiae has been investigated. The synthesis of 4 and 5 coupled with the availability of 3 facilitated a search for these sterols in commercial yeast sterol concentrates, fresh laboratory grown yeast and fresh brewery grown yeast. Sterol 4 was not detected in any of these mixtures whereas 5 was found in the first and last and 3 was present in all three sources investigated. Investigation of incorporation of [2-3H]lanosterol into 3, 4 and 5 revealed significant incorporation into 3 but neither 4 nor 5. This observation suggests the principle pathway for ergosterol biosynthesis initially involved 1 leads to 3 leads to 7. Incubation of a mixture of [2,4-3H]zymosterol and [26,27-14C]lanosterol with S. cerevisiae revealed that during the initial phases of aerobic growth the major route from 7 to ergosterol involves zymosterol (11) but as 11 accumulates 4 alpha-methyl-24-methylenezymosterol (8) assumes equal importance.


Neurological concepts of pain management in head and neck cancer. The physiological basis, historical evolution, and our own practice of stereotactic surgery for the relief of cancer pain are briefly described. Thirty-nine procedures have been performed on 33 patients, 27 for pain due to cancer. Lesions have been made both in the specific neospinothalamic pathway of midbrain and thalamus and in the non-specific reticulothalamic system with an overall five per cent mortality and five per cent morbidity. Initial pain relief was achieved in 90 per cent of patients with relief till death in 74 per cent. Lesions were more successful in releiving pain when the specific pain system was involved but only if the analgesia produced by the lesion "covered" the patient's pain. On the other hand, dysesthesia occurred only with lesions of the specific system, especially if they involved the thalamus.


Atrophic rhinitis: Proplast as an implant material in surgical treatment. The use of Proplast as an implant material is described in the treatment of atrophic rhinitis. It offers an alterantive implant material to bone graft obtained from the patient. There is no resutling morbidity in relationship to a donor site.


Inhibition of oxidative metabolism in Escherichia coli by d-camphor and restoration of oxidase activity by quinones. Oxidative metabolism in whole cells of Escherichia coli strain 82/r was inhibited by d-camphor when glucose, pyruvate, or succinate was used as substrate. Inhibition was not due to lower surface tension in d-camphor-treated cell suspensions nor was it a function of cell permeability. Succinic, lactic, and NADH-oxidase activities were inhibited in alumina powder cell-free extracts (80 mug of protein/ml) by d-camphor (1100 mug/ml). NADH: and succinic: DCPIP oxidoreductase enzymes were unaffected by d-camphor. Menadione (vitamin K3) restored succinic, lactic, and NADH-oxidase activities in d-camphor-inhibited cell-free extracts. Concentrations of menadione used to restore succinic and NADH oxidase activities were not stimulatory in non-camphor-treated extracts. Succinic oxidase activity in d-camphor-inhibited cell-free extracts was also restored by ubiquinone (Q6) but not by vitamin K1. These results are interpreted to indicate that d-camphor may affect quinone function in E. coli.


The influence of hydrostatic pressure upon biochemical activities of heterotrophic bacteria. Heterotrophic terrestrial and marine bacterial protein synthetic rates decrease by about 16% of the 1-atm rate for every 100 atm of pressure increment, within the pressure range of 1 to 500 atm. It is proposed that protein synthesis is nore severely inhibited by pressure than are other anabolic or catabolic reactions.


Physiological and psychological effects of alpha and EMG feedback training with college drug users: a pilot study. Twenty-four volunteer college students who were regular drug users were randomly allocated to three training groups of equal size: alpha feedback, EMG feedback and a joked control group. The subjects were unaware of which feedback condition they received and were asked to practise at home during a six-month follow-up period in order to achieve a relaxed state similar to that experienced during training. No group was successful in retaining gains made in their alpha levels during each session, but the EMG group significantly reduced their muscular activity during training and retained the improvement during follow-up. The alpha and joked groups did not greatly improve their EMG during training but at follow-up achieved the same levels as the EMG group. There was evidence to suggest that a reduction in drug use among light and medium users was maintained during follow-up. Significant and lasting improvements were made by each group in the duration and quality of their sleep and anxiety levels were reduced.


D. Campbell Meyers, M.D., L.R.C.P., M.R.C.S. (Eng.) 1863-1927, pioneer of Canadian general hospital psychiatry. The paper describes the pioneering contributions of Dr. D. Campbell Meyers (1863--1927) to general hospital psychiatry during the first decade of this century at the Toronto General Hospital where, between 1906 - 1911, he operated the first public general hospital psychiatric unit in Canada. His conflict with Dr. C. K. Clarke who opposed his efforts is documented. To honour Meyers' contributions the Department of Psychiatry, Toronto General Hospital, has been named "The D. Campbell Meyers Memorial Department of Psychiatry".


Clinical studies of methanol extraction residue fraction of Bacillus Calmette-Guérin as an immunostimulant in patients with advanced cancer. Forty patients with advanced gastrointestinal cancer, all previous chemotherapy failures, were treated with a methanol extraction residue of Bacillus Calmette-Guérin (MER) alone, administered intradermally in either weekly or every-4-week schedules. The only significant side effect was local cutaneous reaction ranging from papules through pustules to draining ulcerations. On the weekly schedule these reactions frequently reached a point of clinical intolerability. In spite of the advanced nature of their disease, 29% of the patients had increased reactivity to recall antigens; 57% showed an increased reaction to dinitrochlorobenzene challenge during MER therapy; 38% had significant increases in lymphocyte blastogenesis to phytohemagglutinin mitogen, 48% to concanavalin A, and 43% to pokeweed mitogen. Increases (greater than 25%) in immunoglobulins A, M, and G were also observed in 31, 41, and 28% of patients, respectively. MER therapy was associated with increased thymus-derived (T) and bone marrow-derived (B) cells and an increased ratio of T-cells to B-cells. Increases in those determinants reflecting cellular immunity (skin tests, phytohemagglutinin and concanavalin A blastogenesis, and T-cells) showed a positive correlation with patient survival. Increases in those determinants associated with humoral immunity (pokeweed mitogen blastogenesis, immunoglobulins, and B-cells) had, if anything, a negative survival correlation. In comparing administration schedules, the weekly method produced more frequent increases in dinitrochlorobenzene response, more rapid increases with higher peaks in lymphocyte blastogenesis transformation, and more frequent increases in circulating T- and B-cells. The every-4-week method was associated with significantly greater frequencies of increases in immunoglobulins A and M. Of 36 patients with measurable disease, 3 showed greater than 50% objective responses, 2 showed a 25 to 50% response, and 1 showed a mixed response. MER is a potent simulus to cellular and humoral immunity in the patient with advanced gastrointestinal cancer. This stimulation may occasionally result in a clinically evident antineoplastic effect.


Studies in pericardial function. I. Cardiovascular effects of assisted ventilation, thoracotomy and pericardiectomy in the anesthetized dog. In anesthetized dogs, initially breathing spontaneously, the effects of intermittent positive pressure ventilation (IPPV), thoractomy and pericardiectomy on cardiac function, pericardial-ventricular relationships and cardiopulmonary volumes were evaluated. After IPPV stroke volume and work decreased. After thoracotomy there was no further change in cardiac output or work. Pericardiectomy superimposed at this point had no clear-cut effect. End-diastolic transmural pressures (ventricular minus pericardial) and cardiopulmonary volumes were not altered at any stage of the study. In the ventilated open-chest dog the effects of IPPV are greater than those of thoracotomy, and the extent of the effects is related to the level of IPPV. With moderate IPPV and adequate ventilation, the systemic hypotension, marked decreases in cardiac output and reduction of heart volumes reported by previous investigators can be avoided. The removal of the pericardium does not appear to have any deleterious effect in the normal dog.


Glucose dependent alterations of mitochondrial ultrastructure and enzyme content in mouse pancreatic islets maintained in tissue culture: a morphometrical and biochemical study. Isolated islets of Langerhans from mice were maintained in tissue culture for one week at either a high (28 mM) or a low (3.3 mM) extracellular glucose concentration. Electron microscopic morphometry by means of stereological methods revealed a much greater volume of mitochondria in islet cells cultured at low glucose than in those cultured at high glucose. The former islets also showed a higher activity of the mitochondrial marker enzyme, L-3-hydroxyacyl-CoA-dehyrogenase (E.C.1.1.1.35). These results indicate a true mitochondrial hypertrophy at the low glucose concentration. Although it is known from previous studies that the islet cell  metabolism is diminished after low-glucose culture, the present observations of an increased mitochondrial volume probably do not reflect a degenerative process, but rather adaptive changes towards oxidation of energy yielding substrates other than glucose.


Pancreatic islet cytology of Ictaluridac (Teleostei). The endocrine pancreas of the bullhead catfish, Ictalurus nebulosus, and the channel catfish, I. punctatas was studied by light and electron microscopy. In addition to the usual A, B and D cells, a fourth endocrine cell type was consistently observed in the electron microscope. All endocrine cell types were innervated. The vesicles of most of the nerve endings were ultrastructurally different from typical adrenergic and cholinergic vesicles, strongly suggesting the possibility of a third autonomic neurotransmitter serving as a regulator of catfish islet secretion.


Time of fixation and the localization of Gomori-positive and neurophysin-containing structures in the rat hypothalamus. Rats were decapitated and the complete head stored at 4 degrees C. At various time intervals, up to a maximum of 14 days, the brains and pituitary glands were removed and fixed in formalin-picric acid. Neurosecretory material (NSM), as revealed with crotonaldehyde fuchsin, and neurophysin as demonstrated by means of immunoperoxidase histochemistry were localized in the hypothalamic magnocellular nuclei, median eminence and posterior pituitary gland. NSM and neurophysin were present in the paraventricular (PVN) and supra-optic (SON) nuceli up to periods of 14 days although the cellular morphology was disrupted after about 4 days. After 4 days there was an abundance of positive staining "droplets" in both the SON, PVN and along the nerve fibers of the supraoptico-neurohypophysial tract. Pituitary gland NSM and neurophysin were still present 14 days post mortem. The slab gradient polyacrylamide gel electrophoresis pattern of soluble proteins extracted from the posterior pituitary gland from the experimental and normal animals showed almost complete identity confirming the minimal action of proteolytic enzymes at 4 degrees C up to a period of 6 days. It is concluded that as long as the tissue is cooled, rapid fixation of hypothalamic tissue is not essential for the localization of NSM and neurophysin.


Oxytocinase-immunohistochemical demonstration in the immature and term human placenta. Oxytocinase (cystine aminopeptidase) was purified from human retroplacental serum by a combination of fractional precipitation, hydroxylapatite chromatography and gel exlusion chromatography on Sephadex G-200. The purified enzyme possessed a specific activity of 980 mIU/mg using L-cystine-di-p-nitroanilide as substrate. This represented a 3200 fold concentration from the starting material in an overall yield of 12%. Antibodies against oxytocinase were raised in rabbits and the gamma-globulins fraction labelled with fluorescein isothiocyanate prior to its use in the immunofluorescence histochemical localization of the enzyme in human placental tissue. Oxytocinase was confined to the syncytiotrophoblastic cells of normal term, and immature placentas as well as in placentas from patients suffering from severe toxaemia. Specific immunofluorescence was also present in the outer margins of the chorion and to a lesser extent in the amnion.


Types and distribution of lamellar bodies in first cleavage Xenopus embryos. Lamellar bodies in first cleavage Xenopus laevis embryos have been examined with the electron microscope. Based on the arrangement and relative thickness of the lamellae, three types of lamellar bodies have been identified. In the first type, a band of two to ten lamellae (spacing about 3.5 nm) was interwined at random. This type of lamellar body was seen closely associated with the concave or the forming face of the Golgi body and may have been derived from this organelle. The frequent association of these lamellar bodies with the growing cleavage furrow may suggest a transfer of materials from the Golgi body to the furrow through the lamellar bodies or their precursors. In the second type, typical myelin-like figures with a repeating distance of about 10 nm were located close to the furrow tip. In the third type, membrane whorls containing tetrads of lamellae (approximately 16 nm thickness) were seen associated with lipid droplets. If the lamellar bodies observed in this study were formed during fixation then the appearance of three different types after the same fixation procedure would indicate the presence of a similar number of pools of precursor materials which differ in their chemical composition and distribution in the embryo.


Histochemical and electron microscopy investigations on medullary bone. Folliculin administration to pigeons stimulates the development of medullary bone in marrow spaces of the femora and other long bones. It is a specialized osseus tissue not devoted to mechanical functions and which is rapidly reabsorbed before egg-shell formation. Medullary bone is formed and reabsorbed in the same way as other types of bone. Consequently, because of its very rapid rate of formation and resorption, it represents an ideal tissue for studying osteoblastic, osteoclastic and osteocytic activity, and the calcification process. Medullary bone is deeply stained by PAS, Alcian blue and colloidal iron and is metachromatic after toluidine blue staining. This shows that its interfibrially ground substance contains relatively high amounts of glycoproteins and acid proteoglycans. Calcification initially occurs in maxtrix vesicles (or calcifying globules) which are very numerous between the collagen fibrils of the osteoid tissue, and successively spreads into the surrounding interfibrillar matrix. Here, the crystals are closely related to thin, filament-like organic structures which seem to be components of ground substance proteoglycans. These findings confirm that in medullary bone, as in other types of calcifying tissue, the inorganic substances is initially laid down within calcifying globules and is successively closely related to organic, non-collagenous, filamentous organic structures (crystal ghosts) which probably represent a framework for calcium salt deposition.


Studies of mouse mitochondrial DNA in Escherichia coli: structure and function of the eucaryotic-procaryotic chimeric plasmids. The mouse mitochondrial DNA genome has been cloned in Escherichia coli by linking it to the pSC101 plasmid replicon at cohesive-ended cleavage sites generated by Eco Rl restriction endonuclease. The four possible configurations of chimeric molecules that contain the nucleotide sequences of mitochondrial DNA in their native relationship were distinguished by Hind III restriction endonuclease digestion and electron microscopic heteroduplex analysis. Chimeric molecules utilize the pSC101 replication origin and do not maintain the "D-loop" region or the low frequency of ribonucleotides found in native mitochondrial DNA. Hybridization of the RNA synthesized in E. coli minicells carrying the four types of chimeras indicates that transcription occurs predominately on the light strand of the mitochondrial DNA in all cases. This result implies that initiation of RNA synthesis occurs within the mitochondrial DNA segment. Although specific polypeptide synthetis is directed by the mitochondrial DNA segment of each of the chimeras in E. coli minicells, the molecular weight distribution of the polypeptides synthesized consists primarily of low molecular weight species and thus differs from that observed in mitochondria in mouse L cells.


Observer agreement in evaluating coronary angiograms. The reliability of interpretation of coronary arteriography as a diagnostic tool was investigated in a sub-study of the VA Cooperative Study of Surgical Treatment for Coronary Arterial Occlusive Disease. Twenty-two physicians with varying levels of experience read 13 cine angiograms -- blind -- on two different occasions. Analysis of inter- and intraobserver variability showed that angiographic items about which observers were most inconsistent from one reading to the other had the largest interobserver disagreement as well. They were the distal portions of the left anterior descending and left circumflex arteries. Among the items on which there was most consistent agreement -- namely, the right main coronary artery and presence of ventricular aneurysm -- there was most often agreement between observers as well. When individual readers were evaluated, some observers were far more consistent in their own readings of all the angiographic items than others. This intraobserver agreement in turn correlated fairly well with how often they agreed with the other observers and with how much experience they reported having in reading coronary cineangiograms.


Profile of high risk in people known to have coronary heart disease: A review. A review of the current literature of the high-risk factors associated with cardiac morbidity and mortality in patients with established coronary heart disease is presented. Univariate risk factors include age, comorbidity, characteristics of the index coronary event, electrocardiographic findings including ventricular premature beats and response to exercise tests, angiographic severity of the coronary disease, cigarette smoking, psychosocial status, and the chronometric interval after the index coronary event. Multivariate analysis of risk factor combinations is discussed. Risk reduction requires not only an understanding of the clinical course of a disease process, but also, modification of the amenable factors which contribute to excess risk of death.


Time, place, event of sudden death.

Time, place, event of sudden death. Sudden death, known since antiquity, is almost always due to cardiovascular disorder, but not exclusively to coronary artery disease. It is postulated that two processes, action of pre-existing risk factors (enlarged heart, abnormal conduction, extra systoles, ischemia) and new, untoward events (acute infarction or ischemia) culminate in the catastrophic event. Sudden death is apparently most likely during waking hours; sleep is a lesser risk. Which subjects are at greatest risk and which circumstances are most dangerous remain unknown and are important topics for further investigation.


Nasal hyposensitization and hay fever.

Nasal hyposensitization and hay fever. A clinical trial of pre-seasonal hyposensitization with intra-nasal grass pollen extract was carried out in forty-two patients. Results were assessed subjectively, by clinical examination during the pollen season, and by measurements of nasal air flow. Useful relief of symptoms was considered to have occurred in fifteen out of twenty-one patients who had pre-seasonal treatment with pollen extract and in five out of twenty-one patients who had similar treatment with carbol saline as a placebo. Of twelve patients who were symptom free in the pollen season ten had nasal hyposensitization. It is concluded that this simple and safe method of treatment gives results comparable with those of pre-seasonal injections.


Intranasal betamethasone valerate in the treatment of seasonal rhinitis. Betamethasone valerate aerosol given in doses of 100 mug into each nostril twice daily was compared with a placebo in a double-blind, cross-over trial involving thirty patients with seasonal rhinitis. Patients recorded symptoms of eye irritation and watering, sneezing, rhinorrhoea, and nasal blockage, on a diary card. Analysis of the symptom scores showed that nasal symptoms were significantly better on betamethasone valerate than on placebo (P less than 0.01) and that nasal blockage in particular was improved (P less than 0.001). The patients' preference was significantly in favour of the active compound (p less than 0.02) and no side-effects were noted. It is concluded that betamethasone valerate offers a safe and effective form of treatment for seasonal rhinitis.


The effect of beclomethasone dipropionate aerosol on allergen induced nasal stenosis. Nasal resistance to air flow has been used to evaluate the effect of beclomethasone dipropionate aerosol on allergen induced nasal stenosis. Sixteen patients with allergic rhinitis due to pollen were investigated in a randomized double-blind cross-over study with beclomethasone dipropionate aerosol and placebo aerosol. Only patients reacting to challenge were chosen. The study was carried out in the pollen-free season. After 1 week on either active or placebo aerosol a basic resistance value was determined followed by allergen challenge. Nasal resistance was determined 15 min and 7 hr after challenge. The aerosols were changed and after another week the procedure was repeated. There was significant preference (P less than 0.01) for beclomethasone dipropionate aerosol.


Application of pepsin-digested horse antibodies for quantitative immunoelectrophoresis. The majority of the precipitating antibodies in hyperimmune horse serum belong to the beta globulins, as demonstrated by reversed immunoelectrophoresis. As these antibodies migrate in agarose gel during electrophoresis in conventional pH = 8.6-8.9 buffers, horse antiserum cannot be used satisfactorily for quantitative immunoelectrophoresis. On pepsin digestion of horse antiserum F(ab)' 2 fragments are generated which migrate with the gamma globulins. This cleaved product works well in quantitative immunoelectrophoretic techniques.


Influence of adrenaline on the dissemination of antibody-producing cells from the spleen. Plaque-forming cells (PFC) and rosette-forming cells (RFC) were quantificated in splenic venous and splenic arterial blood and in spleen suspensions of guinea-pigs during a secondary immune response to sheep red blood cells (SRBC). The splenic veno-arterial differences in content of PFC and RFC were determined, indicating whether there had been a release of such cells from the spleen into the blood. The effect of an intracardial injection of adrenaline on the release of immune lymphocytes was investigated. In immunized control animals a splenic release of antigen-binding and antibody-forming cells was found, the release being restricted to the peak of the immune response in the spleen. However, after exogenous adrenaline a considerably increased release of both antigen-binding and antibody-forming cells occurred during a longer period of the immune response. Thus, adrenaline caused an enormous release of PFC from the spleen into the blood on day 4 of the secondary immune response, resulting in a diminished number of PFC remaining in the spleen after the treatment. A physiological significance of an adrenaline-induced dissemination of immune lymphocytes in the body during an immune response to a severe infectious disease is suggested.


Nippostrongylus brasiliensis in young rats. Lymphocytes expel larval infections but not adult worms. Young rats were not able to expel adult N. brasiliensis infections even when the worms were damaged by antibodies and the young rats were given all the cellular components (sensitized lymphocytes and bone marrow cells) shown to be necessary for the expulsion of antibody-damaged worms from adult rats. In contrast, most of the worms were expelled from young rats given sensitized lymph node cells on the day of a larval infection. These results show that the reduced ability of young rats to respond to infection by producing sensitized lymphocytes only partly explains their inability to expel the worms. It was not possible to explain the failure of young rats to expel adult worms by hypothesizing that they develop an active factor which prevents the cells from acting on the worms. It is also unlikely that worms persist in young rats because they differ in their susceptibility to cells compared with antibody-damaged worms from mature rats. This work suggests that the immune mechanism which affects the immature stages of this nematode may differ from that which controls the adult stages.


The effect of human alpha 2-macroglobulin on the restoration of humoral responsiveness in x-irradiated mice. The effect of human alpha 2-macroglobulin on the recovery of the humoral immune response in X-irradiated mice has been assessed by measurement of antibody-forming cells in the spleen 5 days after challenge with sheep erythrocytes. The capacity of this protein to promote the recovery of immune responsiveness was variable, and appeared to be influenced by the strain and sex of the mice, and the dose of alpha2M given.


Autoimmunity to sperm antigens in vasectomized men. Sera from vasectomized men were tested for the presence of antibodies directed against sperm antigens. A high percentage (about 55%) of the vasectomized men developed agglutinating antibodies. A lower percentage (22%) also developed low titres of antibodies to human protamine, as detected in the indirect IFT on swollen sperm heads and 22% developed cytotoxic antibodies. A correlation was found between the presence of anti-protamine antibodies and the presence of agglutinating and of cytotoxic antibodies. This correlation, and also the fact that they developed after vasectomy, indicates that the formation of antibodies against human protamine is a result of an autoimmune response to spermatozoa. The indirect IFT was also performed on 'normal' unswollen spermatozoa. All the sera were positive at least on one of the sperm antigens located in the acrosome, equator, or post-nuclear region, but no increase nor decrease in titre was found after vasectomy.


Observations in immunotherapy of lymphoma and melanoma patients. Maintenance of remission solely by repeated BCG vaccinations in seven patients with non-Hodgkin's lymphoma who had achieved a complete clinical remission with initial standard therapy has provided sufficient encouragement to begin a randomized clinical trial. In vitro lymphocyte responses to mitogens and PPD used as parameters of cell-mediated immunity have not proved to be of value in predicting early or late recurrence in six pre-trial and trial patients. Eight out of twenty-one patients with malignant melanoma have shown a satisfactory clinical response (10-34 months) to immunotherapy. Those who respond must show immunological reactivity to the stimulating agent, however the best clinical responses were not associated with the highest degrees of in vivo and in vitro sensitization. The skin reactivity and the in vitro lymphocyte response to PPD as well as a 2-3-fold increase in the appearance of colony-forming units in the peripheral blood following the intratumour injection of BCG or PPD are helpful in prognosis and management of these patients. All patients with malignant melanoma who presented with a PHA response less than 40% of normal made a poor response to immunotherapy. Autopsies performed on seven patients dying with extensive melanocarcinomatous disease failed to show any serious adverse toxic reactions or infections from oral and intratumour injections of BCG.


Reversible granulocyte killing defect in anorexia nervosa. Three patients are described with anorexia nervosa in whom malnutrition was present with neutropenia and a granulocyte bactericidal degect. Their peripheral blood granulocytes were found to have a reduced rate of killing of Staphylococcus aureus and E. coli in vitro. The opsonic activity of the patients' sera towards Staphylococcus aureus was normal. One of these patients had recurrent episodes of infection which stopped after she had gained 13 kg in weight. Clinical recovery was associated with a return of granulocyte function to normal. It is concluded that granulocyte bactericidal capacity towards a variety of bacteria may be reduced in patients with anorexia nervosa who have malnutrition. This type of acquired granulocyte bactericidal deficiency appears to be reversible.


Surface markers on lymphocytes of multiple sclerosis patients. Peripheral blood lymphocytes of twenty-two multiple sclerosis (MS) patients and thirty-five healthy controls were examined for the presence of surface markers characteristic for B lymphocytes (surface immunoglobulin, receptor for C3 (EAC), reporter for Fc (EA) and the spontaneous rosette-forming capacity characteristic of T cells. The results obtained indicate that the number of B and T cells in MS is similar to controls, as evaluated by the presence of surface immunoglobulin and E rosette-forming capacity. However, a statistically significant reduction in the percentage of lymphocytes bearing C3 receptors has been found in MS patients. It might have resulted from a reduction in the lymphocyte population bearing C3 receptor but no surface immunoglobulin. The EA rosette test revealed the greatest difference between the groups. The difference indicated a reduction in the density of the receptor for 7S Fc on lymphocytes from MS patients. The results obtained are consistent with the hypothesis of an immune deficit in multiple sclerosis.


An immunofluorescence study of warts.

An immunofluorescence study of warts. An investigation of the incidence of wart virus-specific antibody and of virion antigens in patients with single or multiple warts taken from different anatomical sites showed that all warts did not contain antigen. The incidence was identical using either rabbit antiserum or human sera known to contain virus-specific antibody. The warts from sole, heel and toe had a much higher incidence of stainable virus antigen. Virus-specific staining was mainly found in the keratinized and granular layers of the wart but occasional synthesis in prickle cell layer was observed. All the patients who had virus antigens in their warts did not have virus-specific antibodies but no patient who had antibody lacked antigen. Of the virus-specific antibodies, the IgM class was predominant; a smaller number also contained virus-specific IgG antibody; none had virus antigens and both classes of virus-specific antibody. A wart cell-specific IgM was also found along with virus-specific antibodies and independent of them. An IgM producing fibrillar Staining in human embryonic lung cells was noted in some sera.