Purification and properties of a citrate-binding transport component, the C protein of Salmonella typhimurium. Salmonella typhimurium was shown to contain a citrate-binding protein (C protein) which was purified to homogeneity from the periplasmic fraction released by cold osmotic shock. The protein is dimeric, has an apparent molecular weight of 28 000 and an isoelectric point of 6.1. Sodium ions were required for optimum substrate binding, however, the divalent cations Zn2+, Mg2+, and Co2+ were inhibitory. The C protein was relatively stable but sensitive to various detergents and chaotropic agents. Approximately one citrate molecule was bound per molecule of protein and citrate binding (Kd = 1-2.6 microM) was strongly competitively inhibited by DL-isocitrate and DL-fluorocitrate but not by other carboxylates. Neither succinate, glutamate, nor acetate were bound to the C protein. No apparent enzyme activity was associated with this protein. A concomitant reduction in the level of binding protein and in citrate transport activity occurred in osmotically shocked cells as well as with L-malate- or succinate-grown cells. Fluorocitrate-resistant mutants were simultaneously defective in citrate transport, citrate binding, and production of cross-reacting material. One transport-defective mutant did produce citrate binding protein.


Purification and Best Department of Medical Research, University of Toronto, Ont., Canada. Factor EF-P is a nonribosomal (soluble) protein of Escherichia coli that stimulates peptide bond synthesis when certain aminoacyl-tRNA analogues are used. The purification of this protein to apparent homogeneity is described here. EF-P has a molecular weight of about 21 000, a Stokes radius of 27 A (1A = 0.1 nm), and a frictional coefficient of 1.48, suggesting an asymmetric structure. By this and a number of other criteria, EF-P is a new factor that controls peptide bond formation during protein biosynthesis.


Aspartate transcarbamoylase: loss of homotropic but not heterotropic interactions upon modification of the catalytic subunit with a bifunctional reagent. The role of conformational changes in the allosteric mechanism of aspartate transcarbamoylase from Escherichia coli was studied by reacting the isolated catalytic subunit with the bifunctional reagent tartryl diazide. Two derivatives differing moderately in substrate affinity were obtained depending on whether the reaction was conducted in the presence or absence of the substrate analogue succinate and carbamoyl phosphate. The modification was not accompanied by aggregation or dissociation. The modified catalytic subunits retained the ability to reassociate with unmodified regulatory subunits and produced hybrids similar in size to the native enzyme. These hybrids were appreciably sensitive to the allosteric effectors ATP and CTP but unlike native enzyme showed no cooperativity in substrate binding. The Michaelis constants of these hybrids for aspartate were intermediate between that of the isolated catalytic subunit and that of the relaxed state. Activation by ATP was caused by a reduction in Km to the value characteristic of the relaxed state whereas CTP inhibited by lowering the Vmax. The properties of the hybrids are strikingly similar to the modified enzyme obtained by Kerbiriou and Hervé from cells grown in the presence of 2-thiouracil. However, the crucial modifications are found in the regulatory subunits of the enzyme studied by these authors whereas they are located in the catalytic subunits of the hybrids reported here. Our results suggest that interactions between the catalytic and regulatory subunits have considerable effects on the state of the substrate binding sites in the native enzyme.


Purification and characterization of membrane-bound fumarate reductase from anaerobically grown Escherichia coli. Fumarate reductase has been purified 100-fold to 95% homogeneity from the cytoplasmic membrane of Escherichia coli, grown anaerobically on a defined medium containing glycerol plus fumarate. Optimal solubilization of total membrane protein and fumarate reductase activity occurred with nonionic detergents having a hydrophobic-lipophilic balance (HLB) number near 13 and we routinely solubilized the enzyme with Triton X-100 (HLB number = 13.5). Membrane enzyme extracts were fractionated by hydrophobic-exchange chromatography on phenyl Sepharose CL-4B to yield purified enzyme. The enzyme whether membrane bound, in Triton extracts, or purified, had an apparent Km near 0.42 mM. Two peptides with molecular weights of 70 000 and 24 000, predent in 1:1 molar ratios, were identified by sodium dodecyl sulfate polyacrylamide slab-gel electrophoresis to coincide with enzyme activity. A minimal native molecular weight of 100 000 was calculated for fumarate reductase by Stephacryl S-200 gel filtration in the presence of sodium cholate. This would indicate that the enzyme is a dimer. The purified enzyme has low, but measurable, succinate dehydrogenase activity.


Elucidation of the quaternary structure of reversibly immobilized alkaline phosphatase derivatives. Escherichia coli alkaline phosphatase has been reversibly immobilized on Sepharose CL-4B through two different methods, both based on a disulfide linkage, under conditions selected to favour the coupling of the enzyme to the solid support through one covalent linkage. The quaternary structure of the reversibly immobilized subunit, produced by dissociation of the matrix-bound dimer, was examined by cross-linking with the bifunctional reagent dimethyl suberimidate. Following release from the solid support, the protein was analysed by sodium dodecyl sulfate gel electrophoresis demonstrating the presence of a sufficient amount of dimeric structures in the immobilized subunit preparation to account for all the enzyme activity observed in this sample. These results suggest that the subunit of alkaline phosphatase may be catalytically inactive. This approach to studying the quaternary structure of immobilized subunit derivatives offers the opportunity to directly determine the homogeneity and structure of matrix-bound 'monomer' preparations and is particularly useful in determining if low levels of catalytic activity observed in some immobilized subunit populations are due to the presence of contaminating oligomeric structures.


The rep mutation. VI. Purification and properties of the Escherichia coli rep protein, DNA helicase III. The protein product of the rep gene of Escherichia coli is required for the replication of certain bacteriophage genomes (phi X174, fd, P2) and for the normal replication of E. coli DNA. We have used a specialized transducing phage, lambda p rep+, which complements the defect of rep mutants, to identify the rep protein. The rep protein has been purified from cells infected with lambda p rep+ phage; it has a molecular weight of about 70 000 and appears similar to the protein found in normal cells. Stimulation of phi X174 replicative form DNA synthesis in vitro was observed when highly purified rep protein was supplied to a cell extract derived from phi X-infected E. coli rep cells and supplemented with replicative form DNA. The purified protein has a single-stranded DNA-dependent ATPase activity and is capable of sensitizing duplex DNA to nucleases specific for single-stranded DNA. For this reason we propose the enzyme be called DNA helicase III. We infer that the rep protein uses the energy of hydrolysis of ATP to separate the strands of duplex DNA; the E. coli DNA binding protein need not be present. The rep3 mutant appeared to make a limited amount of active rep protein.


A novel assay for endonucleases acting at apurinic sites and its use in measuring AP endonuclease activity in repair-deficient mutants of Saccharomyces cerevisiae. A quick and convenient assay for depurination and AP endonuclease activities has been developed. (The term 'AP endonuclease' refers to a nuclease that acts on apurinic and probably apyrimidinic sites on DNA.) It is based on the observation that different topological forms of DNA, such as open circular DNA and covalently closed circular DNA, bind different amounts of the fluorescent intercalator ethidium bromide, and can therefore be distinguished by their fluorescence. This assay has been used to measure AP endonuclease activity in 22 repair-deficient mutants of Saccharomyces cerevisiae. All 22 had normal or nearly normal AP endonuclease activity. The AP endonuclease activity was partially characterized.


Cross sensitivity of mutator strains to physical and chemical mutagens. Ten different mutator strains of Saccharomyces cerevisiae were tested for cross sensitivity to two alkylaitng agents, ethylmethanesulfonate (EMS) and methylmethanesulfonate (MMS), to determine if any of them are defective in the repair systems which normally deal with damage caused by these agents. For one of the mutators, namely mut2-1, it was shown by genetic analysis that mutator activity and MMS sensitivity are both controlled by the same gene. Two mutants, mut2-1 and mut7-1, were found to be sensitive to MMS but normal to ultraviolet and gamma-rays. Another group is represented by mut1, mut6 and mut8 which are not sensitive to any of the mutagens tested so far. Mutator strain mut2-1 was also shown not to be significantly altered for levels of UV-induced forward and reverse mutations. These observations lend support to the idea of multiple repair systems that deal with DNA damage caused by different agents and also show that mutator activity can often result from the loss of normal cellular repair systems.


Reduction of sensitivity to ultraviolet light in excision-deficient Escherichia coli strain WP2 uvrA. Mutants with increased resistance to UV light were isolated from Escherichia coli WP2 uvrA. Most of these isolates were designated as true or partial revertants of the uvrA- mutation. However, one class of mutants showed only a moderate increase in resistance to UV. The gene responsible for this phenotypic change is closely linked to the lac locus and has been designated the suuA gene (suppressor of uvrA).


Lethal zygosis in recombination-deficient mutants of Escherichia coli. Use of nonselective medium for plating cells following mating has revealed that Rec recipient strains of E. coli may be killed as a result of conjugation. Sensitivity of RecA-, RecB-, and RecC- recipients increases with ratio of donor: recipient cells in mating mixtures and with time of mating. A Rec+ recipient shows no lethal zygosis in these experiments performed without aeration. Cell contact does not seem to be responsible for the sensitivity of Rec- strains, since lethality is prevented when cell contact is permitted but DNA transfer is not. Thus, an event(s) occuring subsequent to entry of donor DNA appears to cause lethality in Rec- recipients.


Tryptophan-requiring parental strains yield Salmonella typhimurium x Escherichia coli hybrid recombinants with functional tryptophan operons. Crosses between an Escherichia coli Hfr trp strain and three Salmonella typhimurium F- trp strains produced some trp+ hybrids in which the tryptophan operon is composed of genes from both parental species.


Purification of infectious pancreatic necrosis (IPN) virus and comparison of polypeptide composition of different isolates. Infectious pancreatic necrosis (IPN) virus was partially purified by freon extraction of infected CHSE-214 cells and concentrated by polyethylene glycol (PEG) precipitation of virus from the medium. Both methods resulted in virus concentrates that could be further purified by two CsCl gradient centrifugations with little loss of infectivity. A Recovery of 80 to 100% of the virus infectivity was obtained and over 100-fold concentration of viral infectivity was achieved by these methods. This purification was used to compare 10 isolates of IPN virus with regard to their physiochemical properties by electron microscopy, buoyant density in CsCl, and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of the purified virions. Electron-microscopic observations showed that the virus isolates were identical in that they were isometric, hexagonal in profile, and had a particle diameter of 71 nm. The buoyant densities of the virus isolates in CsCl were found to be 1.33 g/ml. SDS-gel electrophoresis of the virus isolates revealed the presence of three polypeptides of molecular weight 50, 30, and 27 x 10(3) designated as VP50, VP30, and VP27, respectively.


A simple method for the inactivation of St. Louis encephalitis virus preparations for immunofluorescent microscopy. A study was made of different treatments for the inactivation of St. Louis encephalitis virus in smears prepared for immunofluorescence microscopy. Treatment of infected cells with 0.3% betapropiolactone at 56 degrees C for 40 min resulted in an inactivated virus suitable for immunofluorescence studies.


Effects of some platinum IV complexes on cell division of Escherichia coli. A comparison was made of the effects of cis-tetrachlorodiaminoplatinum (IV) (cis-TCDPt), rans-TCDPt), and hexachloroplatinum (HCP) on growth and cell division of Escherichia coli strains D21 and D22. At or below 40 microgram/mL, cis-TCDPt inhibited cell division but not growth, DNA, or protein synthesis, although areas of increased electron density could be demonstrated in treated cells. In contrast, 40 microgram/mL of trans-TCDPt or HCP inhibited growth. Trans-TCDPt-treated cells developed condensed nucleoids; HCP-treated cells showed no obvious cytological changes to correlate with growth inhibition. Combination of cis-TCDPt with nalidixic acid, both at one-half the lowest filament-forming concentrations, resulted in formation of filaments, suggesting an additive effect. Combination of cis-TCDPt followed by ampicillin on E. coli B/r resulted in single bulges near the center of the filaments. Cis-TCDPt could therefore inhibit an initial step in the septation sequence, possibly at the level of the regulation of the hydrolytic enzymes. Whether cis-TCDPt exerts its effect by interreaction with DNA or with a membrane target is still uncertain.


Aged-related physiological studies comparing Candida albicans chlamydospores to yeasts. From electron-microscopical observations, a decreased metabolic activity in 3-day-old Candida albicans chlamydospores was suggested, and progressive deterioration in chlamydospores aged 2-8 months was shown. Oxygen utilization by chlamydospore-pseudomycelium (CSP) preparations was less than that by yeast, while 3-day-old CSP preparations used significantly less O2 than 24-h CSP preparations. Amino acid incorporation was greater in yeast than in CSP preparations. Leucine incorporation by 20-h yeasts was twice that of 5-day yeasts and 5 times that of 20-h and 5-day CSP. Amino acid decarboxylation was similar in yeasts and CSP and was determined by end-product analyses to be via amino acid oxidase. Light microscopy autoradiography of [14C]leucine incorporation demonstrated that the metabolic activity in CSP preparations was due to the young growing tips of the pseudomycelium and not to mature chlamydospores. Yeasts did not take up trypan blue could be stained if first autoclaved or treated with 10% acid or 10% base. Young chlamydospores grown in the presence of trypan blue developed unstained and became permeable to the dye at 2 1/2 days. These data suggest that chlamydospores of C. albicans do not function in the classical role attributed to spores; i.e., mature chlamydospores cannot germinate, but rather age, deteriorate, and die.


Method for the rapid screening of cellulolytic streptomycetes and their mutants. A method for the rapid screening of cellulolytic streptomycetes and their mutants is reported. The technique consists of a plate assay on media containing filter paper fibres as cellulose substrate. The cellulolytic activity is detected and measured by the formation of clearing zones around the streptomycete colonies. The sensitivity of the method is increased considerably by subjecting the plates to an additional incubation period at 43 degrees C in the presence of a buffer at pH 5.3. by replicating these colonies on other Petri plates containing the appropriate media, it is possible to assess rapidly, not only the degree of catabolic repression of the cellulase production by glucose, but also, in a semiquantitative way, the amount of enzymes produced.


A critical analysis of the classifications of non-Hodgkin's lymphomas. The Rappaport classification of non-Hodgkin's lymphomas was proposed almost a quarter century ago, before the advent of modern immunology. This classification, which is based entirely on morphologic features, has proved its clinical usefulness. In light of recent scientific advances, however, its terminology is not appropriate. Five new classifications have been proposed recently, each claiming to have more merit than the others. The purpose of this study is to critically analyze and evaluate these newly proposed classifications to determine which classification is conceptually and scientifically acceptable as well as clinically useful. The results of the study show that there are more similarities than differences among the Rappaport. Lukes and Collins, Dorfman, British, and WHO classifications; the Kiel classification, however, is fundamentally different (Tables 8, 9, 11). None of these classifications can be used in its proposed form. Based on the analysis of these classifications, a compromise working classification is proposed which incorporates the relevant concepts and terminology from the Rappaport, Berard, Dorfman, WHO, and Lukes and Collins classifications (Tables 15, 16). The proposed compromise classification is an attempt to reconcile the various classifications, and to stimulate others to offer modifications which may bring about a final solution to the problem of classification of non-Hodgkin's lymphomas.


Combination chemotherapy of metastatic breast cancer: a randomized trial comparing the use of adriamycin to that of Vinblastine. Two regimens of chemotherapy for metastatic breast cancer were compared in a randomized controlled fashion. Regimen 1 consisted of cyclophosphamide, vinblastine, methotrexate and 5-fluorouracil (CVMF). Regimen 2 consisted of cyclophosphamide, adriamycin, methotrexate and 5-fluorouracil (CAMF). The patient population consisted of both black and white postmenopausal females who had not received any prior chemotherapy. Objective responses were observed in 25/57 patients treated with CVMF and in 28/51 patients treated with CAMF. Neither race nor choice of chemotherapeutic regimen affected prognosis, although there were differences in the pattern of metastatic involvement between the two racial groups. The median duration of survival of patients who responded to therapy has not yet been reached but will be in excess of 12 months.


Ftorafur, adriamycin, cyclophosphamide and BCG in the treatment of metastatic breast cancer. Ftorafur is a 5-fluorouracil analogue which is slowly metabolized to 5-FU, resulting in prolonged therapeutic levels of this latter drug. Ninety-one evaluable patients with metastatic breast cancer were treated with Ftorafur, Adriamycin, cyclophosphamide, and BCG (ACFTOR-BCG), in an attempt to increase the effectiveness of the program or decrease its myelosuppressive toxicity. The results of this trial were compared to those previously reported with the combination of 5-FU, Adriamycin, cyclophosphamide, and BCG (FAC-BCG). Overall objective response rates were 65% and 76% for ACFTOR-BCG and FAC-BCG, respectively. Durations of response were 12 months and 14 months for ACFTOR-BCG and FAC-BCG (p = 0.53). The median survival of responders was 22 and 23.9 months, respectively. Substantial toxicity was observed with Ftorafur: nausea and vomiting severe enough to cause weight loss was observed in a substantially higher fraction of the patients treated with this drug than with 5-FU. Other side-effects, which were not observed with the 5-FU combination, were somnolence, dizziness, personality changes, tremor, ataxia, and confusion. No differences in myelosuppressive toxicity were observed between the two combinations, and the incidence of infectious complications was identical. The combination of Ftorafur, Adriamycin, cyclophosphamide and BCG did not offer any advantages with respect to increased effectiveness or reduced toxicity over the FAC-BCG regimen in breast carcinoma.


Non-Hodgkin's lymphomas: an ultrastructural study correlating morphology with immunologic cell type. Ultrastructural studies were performed on 40 B-cell and 14 T-cell lymphomas of non-Hodgkin's type (NHL). Most B-cell lymphomas were comprised of neoplastic cells with morphologic features compatible with a follicular center cell origin. Dendritic reticulum cells and their desmosome-associated processes, characteristic of germinal centers, were observed in all 11 cases of nodular poorly differentiated lymphocytic lymphoma and in one of two cases of nodular "histiocytic" lymphoma, but were not identified in the lymphomas with a diffuse growth pattern. Desmosomes were observed between dendritic reticulum cells and were not found between lymphoid cells. Large neoplastic cells comprising lymphomas of "histiocytic," mixed lymphocytic "histiocytic," and "undifferentiated" types were characterized ultrastructurally and immunologically as lymphoid cells. Malignant lymphomas of well and moderately well differentiated lymphocytic types (7 cases) revealed B-cell markers, and represented a distinct homogenous group of neoplasms, with electron microscopic features most closely resembling follicular cuff lymphocytes. T-cell malignancies included lymphoblastic lymphomas (3 cases), large cell ("histiocytic") lymphomas (4 cases), lymphoepithelioid cell ("Lennert's") lymphomas (2 cases), mycosis fungoides (3 cases) and diffuse poorly differentiated lymphocytic lymphomas (2 cases). A consistent finding in the T-cell proliferations was the presence of small and/or large lymphoid cells with extremely irregular and/or convoluted nuclei, which occurred in varying proportions and with variable degrees of nuclear complexity. The nuclear irregularity evident in the neoplastic T cells was distinguishable from that observed for lymphoid cells of B-cell lymphomas. In comparing the cytoplasmic features of the T- and B-cell neoplasms ultrastructurally, the only distinguishing feature was the presence of well developed granular endoplasmic reticulum with dilated cisternae, i.e., plasmacytoid features, predictive of a B cell origin.


Non-Hodgkin's lymphoma: a study of the evolution of the disease based upon 92 autopsied cases. The evolution of the disease process in 92 patients with non-Hodgkin's lymphoma has been analyzed by comparing the initial histopathologic material with that obtained at autopsy. The distribution of lymphoma did not differ substantially among the subtypes, but the gross appearance of the lesions was different among certain subtypes, particularly between the "small cell" and "large cell" types. The diffuse lymphomas showed a remarkable constancy of subtype in the same patient, whereas the nodular lymphomas showed a high frequency of change to a diffuse pattern and/or a larger cell type. Diffuse histiocytic lymphomas with plasmacytoid features showed a striking propensity for involvement of the gastrointestinal tract. The occurrence and significance of bizarre pleomorphic giant cells, epithelioid histiocytes, a "starry-sky" pattern, erythrophagocytosis, hemosiderosis and extramedullary hematopoiesis are reported. All patients received anti-lymphomatous chemotherapy and/or radiotherapy. A second malignancy was observed in 8 patients. Thirteen patients showed no residual lymphoma at autopsy. The cause of death was considered to be related to lymphoma in all 92 patients; 67 had infections identified.


Olfactory neuroblastoma (esthesioneuroblastoma): a light and ultrastructural study of two cases. Olfactory neuroblastoma is a malignant neoplasm with a varied biological behavior. Its clinical course is unpredictable and there is no correlation between its microscopic features and biological behavior. The present study deals with light and ultrastructural characteristics of two cases of olfactory neuroblastoma of the nasal cavity. In one patient, the definitive diagnosis was established on the basis of ultrastructural features of the lesion. The most consistent fine structural findings were the presence of intracytoplasmic densecored neurosecretory granules, "true" and "pseudo-" rosettes, and the neuritic processes emanating from the tumor cells. On the basis of their biochemical, histochemical, and ultrastructural characteristics, olfactory neuroblastomas are similar to neuroblastomas arising from the adrenals or sympathetic nervous system. These findings, therefore, support the hypothesis that olfactory neuroblastomas are most likely of neural crest origin and thus belong to a group of neoplasms collectively known as "apudomas" or neurocristomas. The literature review strongly favors combined surgery and postradiation as the most effective treatment of olfactory neuroblastoma.


Histological transformation of non-Hodgkin's lymphoma: a prospective study. Forty-three lymph node biopsies were performed prior to retreatment in 30 unselected patients who had relapsed following chemotherapy for advanced non-Hodgkin's lymphoma of low grade histological type. Eight patients (27%) showed unequivocal evidence of transformation to a high grade variety of lymphoma. These included 4 out of 21 cases originally having had follicular lymphoma and 4 out of 9 cases having had diffuse lymphoma. In 2 further patients with follicular lymphoma, relapse was diagnosed following examination of the bone marrow and in one the tumor had clearly transformed. In 5 of the transformed lymphomas the cell type was predominantly centroblastic, in 2 immunoblastic and in the remaining 2 centrocytic (anaplastic). Five of the 9 cases developing high grade lymphoma have died after a median interval of 5 months from transformation, whereas only 3 of 23 cases showing no change are dead. In 4 patients low grade lymphoma persisted in the bone marrow at the time of nodal transformation. The clinical circumstances at the time of rebiopsy were unhelpful in predicting transformation.


Idiopathic refractory sideroblastic anemia: incidence and risk factors for leukemic transformation. The clinical course of 29 patients with idiopathic refractory sideroblastic anemia studied by us was reviewed. Four patients developed acute leukemia. We were able to find 27 out of 268 cases of idiopathic refractory sideroblastic anemia reported in the literature which terminated in acute leukemia. Nine of these were well described. The overall incidence of acute leukemia is 10%. In an attempt to identify the risk factors for the development of acute leukemia, the clinical and laboratory features of this group of thirteen well-described cases which terminated in acute leukemia were compared to the remaining 25 of our cases which did not undergo leukemic transformation. The patients who died of acute leukemia tended to have a more severe anemia, a lower reticulocyte count, and increased transfusion requirement, and thrombocytopenia. Thrombocytosis appears to be a relatively good prognostic sign.


Rationale and experimental design for the VA Cooperative Study of Anticoagulation (Warfarin) in the Treatment of Cancer. Anticoagulants have been demonstrated to reduce tumor growth in certain experimental animal systems. Inhibition of clot formation interferes with tumor growth and spread while enhancement of coagulation promotes tumor growth and spread. The fact that the coagulation mechanism is commonly activated in human malignancy together with preliminary reports of therapeutic efficacy of anticoagulants suggests that the coagulation mechanism may be of pathophysiologic significance also in the growth of human tumors. A VA Cooperative Study has been established to test the hypothesis that warfarin anticoagulation will modify the course of malignancy in man. The purpose of this paper is to present the rationale and experimental design for this study with emphasis on management of anticoagulant administration in cancer patients. This paper serves as the basis for forthcoming reports of toxicity and therapeutic efficacy of warfarin in human malignancy.


Mesenteric occlusive lesion and ileal carcinoids. Occlusion of mesenteric vessels due to fibrosis and adventitial elastosis complicated ileal carcinoids and resulted in infarction of bowel in two patients. Synovial sarcoma antedated the carcinoid syndrome in one patient who died; carcinoma of the breast was discovered one year after hemicolectomy in the other. This rare mesenteric occlusive lesion is associated exclusively with ileal carcinoids; it is poorly recognized and the mortality is high.


Mesenchymal tumors associated with hypoglycemia: case report and review of the literature. Hypoglycemia secondary to malignant tumors is rare. Mesenchymal tumors of nonpancreatic origin are the most common tumors associated with the hypoglycemia syndrome, and the clinical features of 115 reported cases are reviewed. The major anatomic distributions of the tumors are thoracic (30%) abdominal (65%), and uncommon locations (less than 5%). Approximately 50% of the tumors were resectable (59 patients), and in 60% the surgical procedure was curative. In the remaining 40% local recurrence predominated related to site of tumor and presence of contiguous organ invasion. The application of multimodality adjuvant therapy for hypoglycemia associated mesenchymal tumors should be based on an understanding of the natural history of the tumor.


Granulocytic sarcoma preceding acute leukemia: a report of six cases. An unusual case of granulocytic sarcoma presenting in a pericardial effusion following trauma and preceding acute myelogenous leukemia (AML) by 8 months is presented. Five additional cases of granulocytic sarcoma preceding leukemia collected by the author are also tabulated. Granulocytic sarcoma in a nonautopsy population of myelogenous leukemic patients was found to be 2.9%. When presenting in an extramedullary site, especially preceding peripheral blood and bone marrow manifestations of leukemia, a misdiagnosis of histiocytic lymphoma may result. In questionable cases, other techniques including the naphthol-ASD-chloroacetate stain, touch imprints, immunoperoxidase stain for lysozyme, and electron microscopy should be utilized. Although only a small series, the most recent cases have shown induction/remission and survival characteristics of AML patients without granulocytic sarcoma.


A clinicopathologic study of orbital and adnexal non-Hodgkin's lymphoma. An analysis of non-Hodgkin's lymphoma involving the orbital structures was performed at the University of Iowa between 1937 and 1975. Sixteen cases of primary orbital lymphoma were diagnosed. Histopathologic reclassification according to the Rappaport scheme and the clinical course of each histologic sub-category was described. There were 5 patients with reactive hyperplasia, 2 patients with well-differentiated lymphoid proliferation with Dutcher bodies which were also felt to be reactive, 3 patients with diffuse poorly differentiated lymphocytic lymphoma, 4 patients with nodular poorly differentiated lymphocytic lymphoma, and 2 patients with diffuse histiocytic lymphoma. It was concluded that the Rappaport classification is applicable to orbital lymphoid tumors and that those lymphomas which do present as primary tumors should be staged as one would stage the same histologic category of lymphoma presenting in other sites. Radiation therapy appears to be an effective treatment for local control; however, patients with primary orbital lymphoma should undergo observation for systemic disease similar to patients with lymphoma presenting in other sites. Excisional biopsy is recommended to facilitate precise classification.


Non-Hodgkin's lymphoma and acute myeloblastic leukemia: a report of 12 cases and review of the literature. Twelve cases of non-Hodgkin's lymphoma and acute myeloblastic leukemia or one of its variants are reported. An additional 33 cases from the literature are reviewed. The mean interval between the diagnosis of lymphoma and acute leukemia is 5.2 years. In 5 patients the two diseases occurred simultaneously or within 6 months of each other. All but 10 of the 45 patients received radiation therapy for their lymphoma. Nine patients had either total nodal or total body irradiation or both. Eight patients received chemotherapy alone. No patient was untreated. Survival after the diagnosis of acute leukemia ranged from 3 days to 14 months, with a median of 3 months. Four patients achieved complete hematological remission following antileukemic therapy. Acute leukemia is estimated to occur in patients with non-Hodgkin's lymphoma in New York State with a 37-fold increased frequency over the expected number. Although acute leukemia may occur in a higher than expected frequency in patients with non-Hodgkin's lymphoma because of an increased risk of a second neoplasm in patients with a primary tumor, it seems more likely that the acute leukemia may be related to the radiotherapy and/or chemotherapy administered to treat the lymphoma. Late death from leukemia after chemotherapeutic or radiotherapeutic remission of advanced non-Hodgkin's lymphoma is preferable to morbidity and/or early death from untreated or inadequately treated lymphoma.


Endodermal sinus tumor of the pineal region: case report and review of literature. This is a case report of an endodermal sinus tumor occurring in the pineal region of a 14-year-old boy who died 4 weeks after surgery and irradiation. Germ cell tumors of this type demonstrate a selective overgrowth of yolk sac endoderm associated with extraembryonic mesoblast and arise preferentially in the gonads of young children. This tumor in the pineal region is rare and carries a grave prognosis as in other extragonadal sites. A summary of previously reported cases is also presented.


Esthesioneuroblastoma.

Esthesioneuroblastoma. Esthesioneuroblastoma is an uncommon tumor of neural crest origin arising in the nasal cavity. Since 1966, 97 cases have been reported in the world literature. In this report an analysis is presented of the following aspects of these 97 patients: age and sex distribution, disease staging, treatment results, interval to recurrence, and survival. Staging was according to a system proposed by Kadish et al. There is a bimodal age distribution with peaks in age groups 11-20 years and 51-60 years. Prognosis is favorable in early stage disease (A and B) with 3-year crude survivals of 88.9% and 83.3%, respectively. Stage C patients have a poor prognosis, with 52.9% surviving 3 years. Treatment consisted of radiotherapy, surgery, or a combination of radiotherapy and surgery. In stage A, prognosis is favorable in all three treatment groups with one 1 of 24 patients dying of recurrent disease. In stage B, recurrence rates and 3-year survivals are also similar in the three treatment groups. Thus, single modality treatment is as effective as combined treatment for early stage disease. Local recurrence is the predominant site of failure occurring in 68% of recurrences. The overall crude survival for the 97 patients was 95.4%, while the determinate survival (corrected for intercurrent disease) was 70.8%.


Carcinoma arising in the wall of congenital bile duct cysts. The incidence of carcinoma arising in the wall of the congenital bile duct cysts is much higher than previously assumed. The authors report 4 such cases of primary and secondary carcinomas and review their clinical features through the similar 59 cases in the literature. Of the 63 cases, the average age was much younger, at least several decades, compared with cases of extra-hepatic carcinoma without bile duct cysts. The female-male ratio was 2.5:1. Racial preponderance was also observed, namely, the majority were Japanese. Additionally, many patients previously received various internal drainage procedures, especially choledochocystoduodenostomy. As the treatment, primary excision of the extrahepatic bile duct cyst seems to give the best results because it can avoid ascending cholangitis and prevent development of carcinoma. However, carcinoma still can arise in the intrahepatic bile duct cyst, which cannot be removed at the present time.


Multifocal adenocarcinoma of the jejunum. We report on an 89-year-old female admitted with signs of intestinal obstruction in whom three independent adenocarcinomas of the proximal jejunum were found. Multiple foci of pyloric gland metaplasia, glandular hyperplasia and dysplasia, carcinoma-in-situ, and several varieties of adenocarcinoma were identified on microscopic examination of 14 cm of excised jejunum. Multifocal adenocarcinoma of the small intestine is extremely rare. We are not aware of any case harboring the complex changes described herein.


Pleural effusion in multiple myeloma.

Pleural effusion in multiple myeloma. Two patients with multiple myeloma are described in whom an unusual complication developed: pleural effusion containing myeloma cells. There are 7 previously reported cases of myeloma in the English literature with this type of effusion. Pleural effusion in myeloma may be due to plasma cell infiltration of the pleura, congestive heart failure, pulmonary embolism, nephrotic syndrome, and second neoplasms. In view of these multiple etiologies, diagnostic thoracentesis should be performed in order to treat the effusion appropriately.


A randomized comparison of two dosage schedules of methyl CCNU: three-week versus six-week treatments. Two dose schedules of methyl CCNU were compared for drug effect and toxicity. One hundred thirteen patients were stratified by tumor site, performance status, and prior chemotherapy and randomized to 100 mg/m2 q 3 wk or 200 mg/m2 q 6 wk orally. Response rates were similar (12% vs. 18%, respectively, in the major tumor sub-types studied) and survival was equivalent. Hematologic toxicity, however, was significantly different, with earlier time to the most severe blood count depressions, more frequent occurrence of severe depression, and a larger percentage of patients requiring dosage reduction on the 6-week regimen. We conclude that the 3-week regimen is superior due to its improved tolerance and is recommended especially for combination drug therapy.


CVP-remission-maintenance in stage I or II non-Hodgkin's lymphomas: preliminary results of a randomized study. The effect of adjuvant combination chemotherapy when given to non-laparotomized patients in remission after radiotherapy in stage I or II non-Hodgkin's lymphoma was studied in a prospective randomized multicenter study. Locally extended field radiotherapy was given to a target absorbed dose of 40 Gy in 20 fractions. Fifty-five patients who were in complete remission 6 weeks after conclusion of radiotherapy were randomized to either no further therapy or to 9 cycles of CVP (cyclophosphamide + vincristine + prednisolone). The relapse-free survival at 30 months was 41% for patients without and 86% for patients with adjuvant chemotherapy (p = 0.02). The survival was the same for both treatment arms, being 90% at 30 months. Fifteen patients have relapsed, 14 of them with extensions and 1 with a recurrence within the radiation target volume. Analysis of subgroups showed that adjuvant chemotherapy in the present series significantly prolonged the relapse-free survival in diffuse histiocytic lymphoma.


1-phenylalanine mustard (L-PAM) in the management of premenopausal patients with primary breast cancer: lack of association of disease-free survival with depression of ovarian function. National Surgical Adjuvant Project for Breast and Bowel Cancers. Breast cancer patients participating in a prospective randomized clinical trial who were less than or equal to 49 years of age, had positive axillary nodes, and who received prolonged 1-phenylalanine mustard (L-PAM) as an adjuvant to mastectomy continue (after 4 years) to demonstrate a significantly greater disease-free survival (p = .007) than do patients who received placebo. Benefit was achieved in patients who were less than or equal to 39 years as well as those who were 40-49 years of age. Those in the younger age group showed a greater improvement in disease-free survival at 4 years relative to their controls (32% vs. 69%; p = .01) than did those in the older age group (48% vs. 61%; p = .09). When patients were examined relative to their nodal status, a highly favorable effect was found to have been achieved with L-PAM in those with 1-3 positive nodes (54% vs. 86%; p = .006). Results indicate that both age groups were benefited. When considered over time, they demonstrate that a relatively greater effect was achieved in the younger women. While L-PAM failed to significantly alter the disease-free survival of those with greater than or equal to 4 positive nodes a slightly better effect was achieved in the group less than or equal to 39 years. Since adjuvant chemotherapy has been found to be more effective in premenopausal than postmenopausal women, it has been presumed that decreased ovarian function, as a result of the chemotherapy, is responsible for the findings. To support or repudiate that concept, information regarding serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2), as well as menstrual function, has been obtained from women receiving L-PAM or L-PAM plus 5-FU therapy. In contrast to findings relative to disease-free survival, ovarian function and menses were most affected in patients 40-49 years of age. Amenorrhea occurred in 73% of patients in that age group and in only 22% of those less than or equal to 39 years (p less than .001). Similarly, a significant increase in LH and FSH and a decrease in E2, all indicative of ovarian suppression, was observed only in the older group of patients. Thus, it is concluded that while ovarian suppression may account for some of the adjuvant chemotherapeutic effect in premenopausal women, the dichotomy of findings in younger and older premenopausal women relative to therapeutic response and ovarian function indicates that other factors could be responsible.


Phase I-II study of ftorafur and methyl-CCNU in advanced colorectal cancer. The combination of Ftorafur (NSC-148958) and methyl-CCNU (NSC-95441) was evaluated in 36 patients with advanced colorectal cancer. The principle toxicities encountered were myelosuppression, gastrointestinal, and neurological. There were no complete responses and only 5/34 (14.7%) patients achieved a partial response. Methyl-CCNU and Ftorafur does not appear to be an effective combination in advanced adenocarcinoma of the colon and rectum.


Cyclophosphamide plus 5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide (DTIC) with or without Corynebacterium parvum in metastatic malignant melanoma. One hundred twenty patients with metastatic malignant melanoma were randomized to receive either cyclophosphamide, 600 mg/m2 IV, on day 1 plus DTIC 200 mg/m2 IV days 1 through 5, or the same chemotherapy plus C. parvum 5 mg/m2 IV on day 8 and day 15. Therapy was repeated every 21 days. Although responses were observed in 13.8% of patients on cyclophosphamide plus DTIC versus 25.5% of patients on cyclophosphamide plus DTIC plus C. parvum, the median duration of remission was 15.6 weeks on chemotherapy and 13.0 weeks on chemotherapy plus C. parvum. Furthermore, survival was similar on both regimens (6.1 months versus 5.7 months, respectively). Favorable prognostic factors included metastatic disease confined to skin or lymph nodes (33% responses), performance status greater than 70% (24% response rate), and administration of three or more courses of chemotherapy (31% response rate). The dose limiting toxicity was myelosuppression, which was equal on both regimens. Chills and fever were common in response to C. parvum, and, rarely hypotension, cyanosis, or immune nephritis was observed. The addition of C. parvum to chemotherapy with cyclophosphamide plus DTIC is not recommended.


Carcinoembryonic antigen in carcinoma of the uterine cervix. 2. Tissue localization and correlation with plasma antigen concentration. Immunoperoxidase staining for carcinoembryonic antigen (CEA) was performed on the tumors of 241 patients with invasive carcinoma of the cervix. Positive tissue staining indicative of a CEA concentration of at least 3 microgram/gm was present in 154 tumors (63%) as opposed to 0 of 30 specimens of normal cervix (p less than .001). Plasma CEA values were obtained at the time of tissue staining on all patients. Plasma CEA concentration was related more directly to total tumor burden (tumor CEA content x extent of disease) than to tumor CEA concentration alone. Progressively rising plasma CEA levels predicted recurrent disease in over 80% of patients whose tumors stained positively for CEA. In contrast, serial plasma CEA values correlated positively with clinical disease status in only 28% of patients whose tumors were devoid of CEA. Immunoperoxidase staining of tissue specimens identifies those patients whose tumors contain high levels of CEA and who therefore should benefit most from subsequent plasma antigen determinations.


Effective dose of L-asparaginase for induction of remission in previously treated children with acute lymphocytic leukemia: a report from Childrens Cancer Study Group. L-Asparaginase, in the dose of greater than or equal to 6000 IU/sq m three times weekly, was demonstrated to be an effective agent in reinduction of remissions in childhood leukemia. Four hundred thirteen children with acute lymphocytic leukemia were treated with L-asparaginase. Doses i.m. ranged from 300 to 12,000 IU/sq m. None of the patients had received prior asparaginase therapy. 6-Mercaptopurine was given p.o. concurrently. All of the patients had experienced several previous relapses, and their disease was not responsive to 6-mercaptopurine. L-Asparaginase was found to be effective in reinducing remissions at the following rates: 9.5% for 300 IU/sq m; 35.1% for 3,000 IU/sq m; 53.5% for 6,000 IU/sq m; and 62.5% for 12,000 IU/sq m. The drug was given three times weekly for four weeks. Hypersensitivity reactions occurred in 6.5% of patients.


Production of fibronectin by human epithelial cells in culture. Human epithelial cell lines derived from both carcinomatous and nomalignant tissues were characterized with respect to the presence and distribution of fibronectin by immunofluorescence microscopy. In cell lines derived from nonmalignant tissues or from primary carcinomas, fibronectin was found predominantly in an extracellular matrix. In contrast, cell lines derived from metastatic carcinomas displayed very little or no fibronectin. Metabolic labeling studies indicated that a positive line synthesized fibronectin de novo rather than absorbing the protein from the media. Negative lines neither synthesized fibronectin nor secreted it into the culture fluid, suggesting that they were not producing fibronectin. Evidence is presented that cells in culture change their properties after extensive subculture since a small amount of fibronectin in an extracellular matrix was observed after extensive subculture of two metastatic lines that were originally negative.


Funding impact of the National Cancer Act and beyond. During the seven years following passage of the National Cancer Act of 1971, the appropriation for the National Cancer Institute (NCI) was increased by nearly $700 million. A major effect of the Act has been increased funding for grants-in-aid, which rose from $93 million in fiscal year (FY) 1970 to over $416 million in FY 1978. Grants programs account for over 60% of the total N.  CI extramural research budget and are divided into four broad categories; research; training (including fellowships); cancer control; and construction. For the first 4 years following passage of the Act, funding for all grants programs increased dramatically. However, growth began to slow in 1976, and the deceleration is continuing. Total NCI obligations for FY 1978 increased at a rate of 7% (as opposed to an increase of 20.3% in FY 1975), which merely managed to keep pace with the estimated Biomedical Inflation Factor of 6.8%. Traditional grants have more than doubled in average cost over the past 10 years, a growth attributable to inflation, more sophisticated and expensive equipment and supplies, and, in some cases, more ambitious projects. The principal types of research grants include: traditional, investigator-initiated research; program projects, a team approach directed toward a common goal; and "core" support used to fund administrative and shared equipment costs of cancer centers. In FY 1977, the actual number of traditional grants awarded declined for the first time in 7 years, while the number of applications for both new and renewal grants increased at an unprecedented rate. Coincidentally, the number of traditional grants awarded this fiscal year increased by 4%, enabling the figure to exactly match that in 1978. While support for traditional grants has remained in the forefront of NCI funding, money for program projects and core support has increased at a greater rate in recent years. However, unlike the years immediately following the Act, emphasis is now being placed on core support (which increased by 9.4% in FY 1978) and program project grants (up 5.1%), as opposed to the exploratory grants necessary to initiate cancer centers. Funds allocated for construction in the wake of the Act are now being reduced, as the pace of development of new centers begins to slow. Although the number of grant awards has decreased, young investigators (35 years old or younger) continued to receiving a significant share of NCI funds, and, in fact, are faring better than older investigators in terms of recommendation, previously referred to as "approval," and award rates. Awards to foreign scientists increased steadily after the Act, achieving their greatest dollar increase ever in FY 1977; in FY 1978, however, they declined by more than 6%. As part of the NCI reorganization plan instituted by NCI Director Arthur C...


Chemical characterization of 465 known or suspected carcinogens and their correlation with mutagenic activity in the Salmonella typhimurium system. Since chemicals exhibiting mutagenic activity pose a potential hazard to their users, there is increasing acceptance of mutagenicity testing as an integral part of a premarketing toxicological evaluation of chemicals. In vitro testing has gained much notoriety as quick and relatively inexpensive means to assess the mutagenic potential of chemicals. However, the innovative use of microsomes to simulate metabolism has not changed the fact that in vitro activation cannot duplicate faithfully the metabolism that occurs in vivo. This shortcoming will express itself by the production of false negatives and possibly false positives during mutagenicity screening. This assertion is also borne out by a reanalysis of the ability of known animal carcinogens to cause mutations in the generally recognized premier in vitro system, the Salmonella-S-9 system. Although previous studies have suggested that a high percentage (greater than 85%) of all carcinogens will be mutagenic in this system, with no indication that false negatives are associated with certain chemical types, these findings are of uncertain practical value due to the limited number of chemical types that were considered. An analysis of 465 compounds with known or suspected carcinogenic activity indicates that about 58% have been adequately tested in Salmonella, that the testing has concentrated on certain chemical types and has neglected others, and that some categories of carcinogens exhibit individual correlations that are unsatisfactorily low by any standard. Poorly detected categories of carcinogens include: azonaphthols; carbamyls and thiocarbamyls; phenyls; benzodioxoles; polychlorinated aromatics, cyclics, and aliphatics; steroids; antimetabolites; and symmetrical hydrazines. Nonstandard procedures are necessary to optimize the testing of chemicals that are bactericidal, that are volatile, or that cross-link DNA. False negatives appear to arise for two reasons: an inability to devise an in vitro activation system that can be reliably used in a standard way; and an inability to detect the entire spectrum of mutational events that can lead to the induction of cancer.


Toxicity of intrapleural Bacillus Calmette-Guérin treatment in animals. The toxicity of intrapleural Tice strain Bacillus Calmette-Guérin (BCG) infection was tested in hamsters. Doses above 10(6) colony-forming units caused significant systemic infection, which could be controlled with conventional antituberculosis therapy. Living BCG in the pleural space did not prevent the healing of bronchial or vascular closures after pulmonary resection. Prophylactic intrapleural BCG (10(6) colony-forming units) significantly reduced tumor growth in the lungs of mice following i.v. injection of 5 x 10(5) syngeneic sarcoma cells. These animal experiments suggest that intrapleural BCG may be administered in the pleural space after lung resection in limited doses if followed by a complementary course of antimicrobial therapy.


A Phase 1 study of high doses of aminopterin with leucovorin rescue in patients with advanced metastatic tumors. We have conducted a Phase 1 study of aminopterin (AMT) with leucovorin (LV) in 17 patients. AMT was administered by bolus injection every 7 to 14 days in dosages from 25 to 425 mg/sq m. LV rescue was instituted at 24 hr and continued for 48 to 72 hr. At dosages above 50 mg/sq m, we observed nephrotoxicity defined as greater than or equal to a 25% increase in serum creatinine 24 hr after AMT administration, but its incidence was not strictly dose related. Urinary alkalinization and volume expansion appeared to reduce the incidence of nephrotoxicity. Nephrotoxic drug courses were associated with 24-hr plasma AMT levels [3.6 +/- 2.0 (S.D.) X 10(-6) M] which were significantly higher than nonnephrotoxic courses (1.6 +/- 1.0 x 10(-6) M) (p less than 0.05). In nonnephrotoxic courses, serum elimination pharmacokinetics appeared to be biphasic with a t1/2 alpha of 1.08 +/- 0.01 hr and t1/2 beta of 12.31 +/- 0.06 hr. Systemic toxicity (myelosuppression and mucositis) could be prevented in patients with impaired AMT clearance by the administration of LV at an increased dose rate. In several courses, systemic toxicity occurred in spite of apparently normal plasma clearance, suggesting that 24-hr plasma levels may not accurately reflect intracellular drug effects. Cytokinetic studies on bone marrow aspirates allowed determination of the rescue effect of LV and may prove useful in predicting marrow protection.


A comparative clinical and immunological assessment of methanol extraction residue of Bacillus Calmette-Guérin versus placebo in patients with advanced cancer. Eighty-four patients with advanced cancer refractory to conventional therapeutic modalities were randomly assigned in double-blind fashion to one of three intradermal treatment regimens: "high"-dose methanol extraction residue fraction of Bacillus Calmette-Guérin (MER) (2.0 mg); "low"-dose MER (0.5 mg); or 0.9% NaCl solution placebo. Toxicity, consisting primarily of cutaneous inflammation and ulceration, was limited to patients receiving MER and was most severe with the high-dose regimen. Pretreatment clinical and immunological parameters were comparable between patient groups. Although a significant number of patients had increases in various immune parameters according to the criteria used, there was no appearent advantage to MER given in either dosage schedule compared to placebo. Patient survival was not affected by either MER regimen compared to placebo. This investigation failed to demonstrate any significant clinical or immunological benefit from MER given in two dosage regimens in patients with advanced cancer with the laboratory methodology used and emphasizes the importance of appropriate controls in evaluating immunostimulants in humans.


Concentration of fibronectin in plasma of tumor-bearing mice and synthesis by Ehrlich ascites tumor cells. In the present paper we have studied: (a) the concentration of fibronectin (FN) in plasma and in ascitic fluid of mice at different times after inoculation of Ehrlich ascites tumor cells; (b) the ability of Ehrlich ascites cells to synthesize and release FN; and (c) the localization of FN in Ehrlich ascites cells by immunofluorescence microscopy. It was found that (a) 4 to 5 days after inoculation of the tumor, the plasma concentration of FN was significantly higher [1.7 +/- 0.07% (S.E.) of total plasma protein] than that in the normal control mice (0.8 +/- 0.035); (b) FN is present in the ascitic fluid in all phases of tumor growth; (c) Ehrlich ascites cells cultured in vitro synthesize and release large amounts of FN in the culture medium; and (d) only about 1 to 2% of the tumor cells show a very small amount of FN, and this is mostly in the area of cell-cell contact.


Mutagenicity of the naturally occurring carcinogen cycasin and synthetic methylazoxymethanol conjugates in Salmonella typhimurium. The aglycone methylazoxymethanol of the naturally occurring carcinogenic glucoside, cycasin, has previously been shown to be mutagenic, but cycasin per se has not. In this work, cycasin was demonstrated to be mutagenic using a modification of the Ames Salmonella test in which it was preincubated with beta-glucosidase and the tester strain in liquid medium. The mutagenicity of cycasin to six histine-depedent Salmonella strains varied considerably with strain HisG46 being the most susceptible. Methylazoxymethyl-beta-D-glucosiduronic acid, which also is nonmutagenic per se, similarly became mutagenic when preincubated with beta-glucuronidase. Methylazoxymethyl acetate, which is slightly mutagenic by the Ames standard pour plate method, became highly mutagenic on preincubation. The mutagenicity of free methylazoxymethanol was confirmed, and a linear dose-response relationship was observed. The common conditions required for activation of nonmutagenic methylazoxymethanol conjugates, the glucoside cycasin and methylazoxymethyl-beta-D-glucosiduronic acid, are 90-min preincubation at 30 degrees, pH 6.5, with an appropriate hydrolase and Salmonella typhimurium HisG46.


Phase I evaluation of tetrahydrouridine combined with cytosine arabinoside. In conventional clinical use, cytosine arabinoside (ara-C) is rapidly deaminated by pyrimidine nucleoside deaminase to the nontoxic compound uracil arabinoside. Tetrahydrouridine (THU) effectively inhibits this enzymatic degradation but is by itself nontoxic. This study demonstrates that concomitant administration of THU markedly increases the myelosuppressive potency of ara-C. When 25 or 50 mg/kg of THU iv and 0.1--0.2 mg/kg of ara-C iv are given daily x 5 days, they produce moderate-to-severe leukopenia and mild-to-moderate thrombocytopenia. A dose of 25 mg/kg of THU with 0.1 mg/kg of ara-C iv daily x 5 days appears appropriate for phase II studies; it produces myelosuppression equivalent to that produced by 3 mg/kg/day x 5 days of ara-C alone. No toxicity occurred with this combination that would not have been expected from ara-C given alone in an equitoxic dose. Although THU and ara-C produced a reduction in peripheral blood and bone marrow blast cells in eight of nine patients with acute leukemia, bone marrow remission did not occur in any of these heavily pretreated patients.


High-dose BCNU therapy with autologous bone marrow infusion: preliminary observations. Nine patients with solid malignancies and extensive prior treatment received high-dose BCNU therapy (600--750 mg/m2) with autologous bone marrow support; following this treatment hematopoietic recovery was studied. The only significant nonhematopoietic toxicity was a probable case of BCNU-induced pulmonary toxicity in a patient who had received massive amounts of prior chemotherapy and chest irradiation. The marrow aspirations prior to cryopreservations had revealed a hypoplastic marrow in four of nine patients. Despite using marrow exposed to prior chemotherapy, neutropenia beyond Day 40 after BCNU therapy was not observed in any patient. One patient did not develop neutropenia of less than 1.5 X 10(9) cells/liter and five patients did not develop neutropenia of less than 0.5 X 10(9) cells/liter. A partial response was observed in one patient and less than partial responses were observed in two other patients. Autologous bone marrow infusion may modify the neutropenia of high-dose BCNU therapy.


Normal metyrapone response after 1 month of high-dose methylprednisolone in cancer patients: a phase I study. A dose of 125 mg of methylprednisolone was given iv each morning for 28 days to six patients with cancer. The posttreatment mean AM/PM serum cortisol (hydrocortisone) level of 19.55/11.27 microgram/dl showed a statistically significant (P less than 0.05) diurnal rhythm and did not differ from the pretreatment level (19.28/11.85 microgram/dl). The response to metyrapone (750 mg orally, every 4 hours X six doses) was assessed in five of the six patients before and after treatment. No abnormally low Compound S (11-deoxycortisol) levels were observed. The mean serum Compound S level (15.30 microgram/dl) achieved after treatment did not differ significantly from the pretreatment mean level (20.24 microgram/dl, P greater than 0.1). No adverse clinical effects were observed during or after this steroid treatment. Since significant hypothalamic-pituitary-adrenal suppression has not been demonstrated, this regimen appears to be safe for use in controlled studies of clinical efficacy.


Mass-spectral studies of isomeric D-ribofuranosylribitol disaccharides from the capsular polysaccharides of Haemophilus influenzae type b and Escherichia coli K 100. Two isomeric-D-ribofuranosylribitols, derived from capsular polysaccharides of Haemophilus influenzae type b and Escherichia coli K 100, were methylated or acetylated, and the products analyzed by gas-liquid chromatography-mass spectrometry. The marked difference in the mass spectra of the methyl ethers of these disaccharides allowed clear distinction between 1- and 2-O-D-ribofuranosylribitol was characteristic for this disaccharide; its isomer, the (1 leads to 2)-linked species, has a base peak at m/e 57. The difference in the base peaks is attributable to fragmentation of the methylated ribitol, as both spectra display common ions characteristic of the methylated D-ribofuranosyl group. For the acetylated disaccharides, the mass spectra displayed common ions characteristic of the acetylated D-ribofuranosyl group. However, no ions similar to those found for the methylated ribitol allowed ready differentiation between the two acetates. Instead, their spectra displayed similar ions, differing somewhat in relative abundance; the M-1 ion, m/e 577, was obtained for both. Comparison of the relative abundance of m/e 139, 259, and 303 in the spectra of the two acetates did allow distinction between them.


A new method for the estimation of cell cycle phases. A method is described, which is applicable to cell renewal systems with an anatomical structure in which all cell locations may be uniquely mapped. Its use is demonstrated on the rat incisor inner enamel epithelium, which forms a one cell thick column in the sagittally sectioned tooth. Cells born in the apical part of the column migrate toward the distal end of the tooth, where they mature. As the cells migrate along the column, they traverse the various cell cycle phases. The present study has been designed to estimate the probability of a cell being in a given phase; all cells touching the basement membrane were numbered, and the number of cells separating any two cells was taken as a measure of distance. Since generally all cells move in one direction (lateral cell migration may occur), it is possible to solve the problem with the aid of functions describing the renewal counting stochastic process in which cell distance serves as an independent variable. The method predicts labelled cell and mitotic rates which agree with those estimated in the usual way. It was then utilized to estimate the fraction of cells in G2.


Localization of cells producting thyroid stimulating hormone in the pituitary gland of the domestic drake. Cells binding anti-bovine TASH beta serum were found exclusively in the rostral lobe of the adenohypophysis of the drake using the peroxidase-antiperoxidase complex unlabelled antibody method. The specificity of the binding of the anti-serum to TSH cells was established by relating the morphology and relative abundance of immunochemically stained cells to the TSH content of the adenohypophysis after experimentally altering the activity of the pituitary-thyroid axis. The TSH activity of the adenohypophysis was assessed indirectly, by the weight of the thyroid glands, and directly, by bioassay. As determined by bioassay, the TSH content of the rostral lobe of the adenohypophysis was much greater than that of the caudal lobe. Compared with control drakes, immunochemically stained cells in birds fed a goitrogen, methimazole, seemed to be enlarged and were closer together, while the stained cells in drakes injected with thyroxine were shrunken and less intensely stained. The TSH content of the adenohypophysis was increased in drakes fed methimazole. Castration did not alter the TSH content of the adenohypophysis or change the morphology of immunochemically stained cells. These observations suggest that in the drake: 1) anti-bovine TSH beta serum binds specifically to TSH cells; 2) the TSH cells occur in the rostral and not in the caudal lobe of the adenohypophysis; and 3) the activity of TSH cells is not inhibited by the feedback effects of gonadal steroids.


Effects of isoproterenol on the dense core and perigranular membrane of atrial specific granules. Following subcutaneous injections of isoproterenol hydrochloride (ISO), atrial cells present a large number of partly degranulated or completely clear "specific granules" enclosed by an intact membrane. Such profiles were never encountered in normal controls and might suggest ISO-induced release of a secretory product. Permeability of perigranular membrane was tested using the extracellular macromolecular tracer horseradish peroxidase (HRP). Reaction product was entirely absent within granules of atrial cells in which the sarcolemma was made permeable to HRP molecules by the ISO injections. This seemed to be the case even in heavily labelled cells in which the peroxidase had penetrated the mitochondrial membranes. In atrial cells impermeable to the tracer, the specific granules closely apposed to the sarcolemma were always HRP-negative. The release mechanism of a possible secretory substance from the specific granules is discussed.


Cyclic changes in the ovary of the rainbow trout, Salmo gairdneri, with special reference to sites of steroidogenesis. In the maturation cycle of ovaries of cultured rainbow trout, three periods can be distinguished: (1) a period of ovulation and previtellogenesis (January-May), (2) a period of exogenous vitellogenesis (May-November/December), and (3) a period of maturation of oocytes (November/December-January). Enzyme cytochemical and electron microscopical data indicate that stroma cells (i.e., interstitial cells and special theca cells) and granulosa cells represent sources of steroids. Steroidogenesis in stroma cells is found throughout the annual cycle, reaching a peak activity in January and February. Weak steroidogenic activity is observed in the granulosa cells of exogenous vitellogenic follicles and young postovulatory follicles. Possible functions of steroids secreted by stroma cells and granulosa cells are discussed.


Peptide hormone-like immunoreactivity in the gastrointestinal tract and endocrine pancreas of eleven teleost species. The distribution of peptide hormone-like immunostaining in the gastrointestinal tract of 11 teleost species was investigated by immunofluorescence. Cells immunoreactive for somatostatin were found in the glandular epithelium of the stomach of four species and in the epithelium of the pyloric appendage of one species. The mid-gut epithelium contained cells reactive with antibodies to glucagon (three species), gastrin (five species), pancreatic polypeptide (five species), and substance P (two species). Cells immunoreactive for met-enkephalin were found in the epithelium of both the mid-gut and the stomach of six species. In six species in which the endocrine pancreas was investigated, insulin-, glucagon-, and somatostatin-like immunoreactivity was observed. Pancreatic polypeptide was definitely localised by immunostaining in cells of the endocrine pancreas of only one out of three species examined. Vasocative intestinal polypeptide-, neurotensin-, bombesin-, and enkephalin-like immunoreactivity was identified in the gastrointestinal nerve fibres in various species. In view of the considerable species variation found, caution should be exercised in generalising about the peptides present in the gastrointestinal tract of fish.


Non-random nature of 2-acetylaminofluorene-induced alterations of DNA template capacity. Male rats were fed a diet containing 0.03% (w/w) 2-acetylaminofluorene (AAF) and their hepatic DNA was isolated and transcribed with E. coli RNA polymerase. Ingestion of the carcinogen-containing diet for 4 days substantially reduced the template capacity of the isolated DNA. This reduction in template capacity was due to an apparent decreased RNA chain size (up to 50%), with no significant changes in initiation or re-initiation of RNA synthesis. This premature termination of RNA synthesis was accompanied, in some instances, by a reduced rate of RNA chain elongation. When the rats were returned to a basal diet for 7 days following 4 days of AAF ingestion, template capacity and RNA chain size returned to control values. Fractionation of hepatic chromatin on a glycerol gradient revealed that inhibition of DNA template capacity occurs on portions exhibiting characteristics of expressed, as well as those with characteristics of repressed, segments of the genome. In contrast, the DNA isolated from a small, highly condensed chromatin fraction (15% of total chromatin-DNA) showed no significant reduction in total template capacity. Analysis of the fidelity of RNA synthesis on this DNA template was performed by determining the rate of addition of individual nucleotide triphosphates to a growing RNA chain. Large reductions in the rates of adenosine and uridine polymerization were observed while no changes in guanosine or cytidine polymerization were found. This suggests the presence of functionally significant carcinogen-induced modifications of adenine. The inhibition in the rate of adenosine and uridine polymerization was reversed when the animals were placed on a basal diet after AAF ingestion.


A nitroindanedione mast cell stabiliser in the treatment of ulcerative colitis: a controlled trial. An orally absorbed mast cell stabiliser (BRL-10833) has been compared with placebo in a double-blind controlled trial in twenty-five patients with ulcerative colitis; patients received each treatment for one month. There was no significant clinical improvement in symptoms or sigmoidoscopic findings with the drug but there was significant improvement in the histological appearance of rectal biopsies during active treatment.


Incidence of immediate sensitivity to Aspergillus fumigatus in a North American asthmatic population. Allergic bronchopulmonary aspergillosis might be less frequent in North America because the incidence of immediate sensitivity by asthmatics to A. fumigatus is less. In order to check this hypothesis, 200 asthmatics were skin tested with two extracts of A. fumigatus which had been shown to produce positive reactions in fifty patients who had allergic aspergillosis. Of the asthmatics, 21.5% reacted to the commercial extract by prick testing and 39% by intradermal testing. Using an extract kindly provided by Professor Pepys, 19.5% reacted to a concentration of 1 mg/ml and 31.5% to 10 mg/ml. By the prick method, 21.5% reacted to both extracts. Specific IgE was measured with one of the extracts and a good correlation (r = 0.48) was found with the size of the prick reaction. The increase in specific IgE was reflected in the increase of total IgE (r = 0.84). The authors conclude that the incidence of immediate sensitivity to A. fumigatus in asthmatic patients in North America is at least equal to that found in the U.K.


[Changes in fluorescent antibodies in experimental bilharziasis with S. mansoni and S. haematobium in mice. Comparison of the results obtained using homologous and heterologous antigens]. Fluorescent antischistosomiasis antibodies appear earlier in mice infected by 150/200 cercariae than in those who only received 20. The use of a homologous antigen in mice infected by S. mansoni or S. haematobium allows earlier and more important responses. The infections by S. haematobium seem to determinate in mice a more important immune responsiveness. Is it a specific parasitic manifestation (S. mansoni being better adapted to rodents?).


Selective inhibitory effect of porcine follicular fluid on follicle stimulating hormone secretion in anterior pituitary cells in culture. Incubation of rat anterior pituitary cells in culture with porcine follicular fluid (treated with charcoal to remove steroids) led to a marked inhibition of spontaneous FSH release while no effect was observed on basal LH secretion. The inhibitory effect of follicular fluid was, however, less specific on LHRH-induced gonadotropin release. Although more potent on FSH than LH release, a significant inhibition of LH release induced by the neurohormone was also observed. This inhibitory activity is mainly associated with material of MW greater than or equal to 10,000 daltons.


[Schistosoma mansoni and haematobium antisera tested by homologous and heterologous antigens. Comparison of the results obtained by indirect immunofluorescence technic]. The study of anti Schistosoma mansoni and anti Schistosoma haematobium sera by homologous and heterologous antigen shows that Schistosoma haematobium has an immunogenic strength as good as that of Schistosoma mansoni. The joint use of the two antigens can be useful in epidemiologic investigations in Africa so as to track down seats of Schistosoma mansoni and Schistosoma haematobium and the prevalence of the infection.


Comparison of long-acting analogues of luteinizing hormone releasing hormone in man. Currently, LHRH, when used therapeutically, is given by parenteral injection every 8 h. We have looked at the release of LH and FSH induced by five analogues of LHRH and compared this with gonadotrophin release after synthetic LHRH. The analogues were substituted in position 6 or in positions 6 and 10 and were given intravenously, intranasally or subcutaneously in three separate studies. After intravenous administration of 100 micrograms, all analogues caused greater release of LH and FSH than did synthetic LHRH. Given intranasally in a dose of 500 micrograms, three of the four analogues tested caused greater LH and FSH release than did LHRH. With tryptophan substitution in position 6 (D-TRP6-LHRH), mean LH levels in five subjects were still above the normal range 24 h after a single intranasal dose. The intranasal administration of selected analogues of LHRH has great potential in the treatment of conditions associated with deficient gonadotrophin secretion, provided that pituitary overstimulation, which may eventually lead to a decrease in LH and FSH output by the anterior pituitary, is avoided.


Oestrogen modulation of gonadotrophin and prolactin release in women with anovulation and their responses to clomiphene. An LHRH test was performed before and at both 44 and 92 h after the administration of 2.5 mg oestradiol benzoate in eleven patients with hyperprolactinaemia, eight with idiopathic secondary amenorrhoea and seven with oligomenorrhoea. The basal serum hormone concentrations and the responses to LHRH were compared with the same tests performed on ten normal subjects during the early follicular phase of their menstrual cycles (days 4--6). Mean basal concentrations of oestradiol in each group of patients and oestrone in those with hyperprolactinaemia were significantly lower than in the normal subjects. The mean concentration of prolactin in women with secondary amenorrhoea remained lower than in the normal women throught the tests (P less than 0.05). The LH and FSH responses to LHRH before oestrogen in patients with hyperprolactinaemia and of FSH in those with secondary amenorrhoea, were greater than in the normal subjects (P less than 0.001). After oestrogen treatment the responses were similar in all groups except in those with oligomenorrhoea where LH and FSH responses at 44 h (P less than 0.05 and P less than 0.01 respectively) and LH responses at 92 h (P less than 0.01) were lower than in normal controls. The responses at 92 h in all groups were greater than at 44 h (amplification) but the amplification at 92 h and at 44 h compared to the pre-treatment responses, tended to be lower in each group of patients compared to the normal controls. In the hyperprolactinaemic group of patients there was a negative correlation between the basal prolactin concentration and the gonadotrophin amplifications at 92 h (P less than 0.01), and a positive correlation between the basal oestrone levels and the amplifications at 92 h (P less than 0.01). The results of the oestrogen amplification test in eleven of the non-hyperprolactinaemic anovular patients were compared with the ovulatory response to 100 mg clomiphene given for 5 days. Six showed a normal oestrogen amplification and they all ovulated. Two patients failed to show greater amplification at 92 than at 44 h and required human chorionic gonadotrophin (HCG) as well as clomiphene to ovulate. The other three showed a diminished LH amplification at 92 h; they required 200 mg clomiphene and showed a prolonged follicular phase. The responses of the hyperprolactinaemic patients to clomiphene were poor and there was a negative correlation between prolactin concentration and oestrogen production (P less than 0.01). All ten hyperprolactinaemic patients treated with bromocriptine ovulated and eight conceived. The oestrogen amplification test appears to have some value in predicting the subsequent response to clomiphene in non-hyperprolactinaemic anovular women.


Immunological changes observed in indeterminate and lepromatous leprosy patients and Mitsuda-negative contacts after the inoculation of a mixture of Mycobacterium leprae and BCG. This investigation was carried out to study the possibility of eliciting favourable immunological changes in small groups of Mitsuda-negative patients with indeterminate leprosy, lepromatous patients who were bacteriologically negative after prolonged treatment with sulphones, and in Mitsuda-negative contacts by means of stimulation with a mixture of autoclaved tissues from Mycobacterium leprae-infected armadillos and living BCG. A radical change was observed in the specific immunological activity of the indeterminate group, all of whom initially had occasional bacilli in cutaneous nerves in biopsies taken from hypopigmented spots, and in the persistently Mitsuda-negative contacts. The 48 hr and 30 day reactions to lepromin, the 48 hr reaction to supernatant antigen from lepromin, the test for bacillary clearence and in vitro lymphocyte transformation (LTT) to M. leprae from human and armadillo lesions all became positive. Of the lepromatous patients studied, only one became positive to all the criteria mentioned above. In the others, the 48 hr reaction to supernatant antigen, the LTT to antigen from a humn source, and the clearance test remained negative, while the Fernandez and Mitsuda reactions became positive. These results are discussed in terms of the possible use of this stimulation procedure in the prevention and immunotherapy of leprosy.


Affinity for measles virus anti-haemolysin of a residual immunoglobulin M in sera of some patients with multiple sclerosis. HEp2 cells persistently infected with measles virus were treated with trypsin to remove haemagglutinin (HA) and examined unfixed or fixed in acetone by fluorescent antibody methods, comparing those sera specific for structural antigens of the virus. Staining patterns combined with the blocking of specific immunofluorescence indicated that IgG specific for measles virus haemolysin could be recognized in multiple sclerosis (MS) sera and that in some sera from which rheumatoid factor had been removed, a residual IgM (MS-IgM) was absorbed to measles virus-infected cells and showed the same specificity in blocking tests as measles virus anti-haemolysin. MS-IgM could be removed from sera by absorption with latex particles coated with human IgG and would seem to be anti-globulin with preferential affinity for anti-haemolysin.


Interactions between IgM antiglobulins and IgG antinuclear antibodies. Some aspects of D-penicillamine activities. In this report we describe an in vitro masking action of IgM rheumatoid factor (IgM-RF) towards IgG antinuclear antibodies (IgG-ANA) which can be recovered by using D-penicillamine (DP) as an unmasking agent. The mechanism of this masking effect was elucidated by using smears of rat free nucleus as substrate, instead of the classical rat liver cryostat sections technique. It was postulated that the 'inhibition' of IgG-ANA by IgM-RF may be due to the formation of high molecular weight complexes (HMWC); this would be the same in vivo. Allowing the formation of HMWC which can be removed from the circulation, IgM-RF may have a protective effect by preventing or minimizing systemic lesions. Therefore, IgM-RF may be considered as a defence response against potentially noxious ANA or antigen-antibody complexes. Induced nephropathy and the high frequency of detection of ANA observed in rheumatoid arthritis (RA) patients treated by DP may be the result of the dissociating effect of this drug on HMWC, in which the activity of pre-existing ANA is hidden when rat liver sections are used for its detection.


The effects of Brescia-Cimino fistulas and cardiac function in transplanted pediatric patients. The cardiac hemodynamic effects of patent Brescia Cimino fistulas were assessed using non-invasive techniques in 10 pediatric patients who had undergone successful renal cadaveric transplantation. All had hemoglobin levels above 11 g/dl and had serum creatinine values of less than 1.5 mg/dl. Studies were done prior to and after temporary fistula occlusion to assess the effect of fistula patency on parameters of cardiac function. Values obtained prior to fistula occlusion were compared to those obtained in a normal control population. It was noted that though fistula patency did significantly alter cardiac function, changes were such that fistula patency could be safely maintained. It was also noted that the difference in cardiac index prior to and after occlusion provided a good assessment of fistula flow.


Membranous glomerulonephritis in congenital syphilis. The glomerulonephritis of congenital syphilis is thought to be immune-complex mediated. We describe a case of C. S. in whom the only manifestation was a glomerulonephritis. Immunopathogenic studies allowed the identification of treponemal antigen in glomeruli. The elution of immunoglobulins from renal tissue showed that glomerular deposits contained antitreponemal antibodies. Cryoglobulins with antitreponemal activity were also isolated from the patient's serum.


The bone inductive capacity of various bone transplanting materials used for treatment of experimental bone defects. The bone defect repairing capacities of autogenous cancellous bone (ACB), decalcified bone (DB), autodigested "low-antigenic" bone matrix (LBM) and preserved bone (PB) were tested in experimental defects in rabbit scapulae. The healing of the defects was most effective with ACB transplants as judged by roentgenologic and microscopic studies. The DB and LBM transplants caused states of healing comparable to each other, and the PB transplants showed the least satisfactory results in this experiment. The ACB, DB and LBM transplants caused bone induction, which resulted in the formation of new bone, subsequent resorption of the transplanted material and rapid reshaping of the callus. The PB transplants caused no bone induction; only some growth of callus on the edges of the defect was observed and pieces of the transplanted material were seen as late as 10 weeks after the operation as dead particles encapsulated by fibrous tissue. The infiltrations of inflammatory cells around the PB transplants indicated a considerable host versus graft reaction whereas this reaction was less with the other materials tested.


Lumbosacral fusion using internal fixation with a spinous process for the graft: a review of 50 patients with a five-year maximum follow-up. During a 5 year period 50 patients with chronic low back pain, with or without sciatica due to a proven lumbosacral disk lesion, underwent a spinal fusion using the Boucher method of screw fixation of the facet joints. The L-4 spinous process was excised and fashioned into an H-shape to act as the bone graft between the spinous processes so avoiding an iliac crest incision. Thus blood loss and operating time were substantially diminished. The screws maintain fixation for an 8-week period during which time the graft is "taking." Being inserted with spinous processes distracted about 1 cm the screws and the graft take the load off the painful posterior part of the annulus, and in addition, the distraction of the spinous processes enlarges the intervertebral foramina by about 1 cm thus making subsequent foraminal stenosis unlikely. Loosening of the screws after incorporation of the graft was observed in some cases, but was not considered to be clinically significant. Spinal fusion using screws for internal fixation including indications and contraindications requires careful consideration of operative technique, postoperative management and complications. The clinical evolution of 86% of the patients was rated good or fair from a clinical standpoint; 86% showed a fusion; 14% demonstrated a pseudarthrosis. Three of the 7 pseudarthrotic patients had a good clinical result. The other 4 had only a fair clinical result. Nine of the 14 patients with fair clinical results had a fusion radiologically; all 7 of the clinically unimproved had a fusion. Since the graft is placed at least 1 cm dorsal to the laminae, spinal cancal stenosis is extremely unlikely.


A new surgical approach in the treatment of depressed lateral condylar fractures of the tibia. Surgical repair of 18 fractures of the laterotibial condyle was performed by a surgical technique combining features of several standard procedures. Complete lateral exposure of the tibial articular surface is obtained through resection of its fibular head. Anatomical reduction is obtained under direct vision. The lateral meniscus is usually preserved and bony reduction is maintained by cancellous and cortical bone graft reinforced with internal fixation.


Degradation of bone matrix morphogenetic activity by pulverization. The yield of new bone from implants of pulverized demineralized whole matrix and bone matrix gelatin declines as the particle size decreases in diameter below 125 microns. The corresponding increase in surface area and mechanically-induced free radicals is associated with an increase in solubility of bone matrix proteins. These changes in physiocochemical properties and the concomitant reduction in bone yield suggest that prolonged pulverization denatures a bone morphogenetic protein (BMP).


Experimental reaming of the proximal femur and acrylic cement implantation: vascular and histologic effects. The technique of microangiography and correlated histology was used, in experiments on the proximal femora of mature mongrel dogs, to study the effects of simple medullary reaming and of reaming followed by insertion of acrylic cement. The hip joints were not disturbed. A segment of Steinmann pin, representing the stem of a hip prosthesis, was inserted into the cement as it was polymerizing. The most significant results, after reaming alone, were devascularization and apparent necrosis of large areas of the cortex of the subtrochanteric femoral diaphysis. Full recovery was observed in 6 months. However, when acrylic cement was introduced following the reaming, extensive necrosis of the inner layers of the diaphyseal cortex was still present after a year, when the observations were terminated. The presence of necrosis of bone was judged by the lack of visible osteocytes in the lacunae. After the devascularization caused by filling the diaphyseal medulla with acrylic cement, the apparently necrotic cortex was revitalized without osteoclasia. That is in contrast to the results in another study in which osteoclasia and osteoneogenesis were required to restore cortex with empty cell lacunae. This raises the question of dormant osteonecrosis in contrast to total osteonecrosis. These studies show, in thin histologic preparations, acrylic bone cement in situ in the tissues. A fibrous membrane was found at the cement-bone interface in most areas. It was thicker in the diaphysis than in the metaphysis. The physiologic effects of acrylic cement implantation are considered to be likely factors in the loosening which is observed clinically when the femoral components of hip prostheses are under protracted stress.


Clinical pharmacokinetics of phenytoin.

Clinical pharmacokinetics of phenytoin. Phenytoin is a relatively insoluble weak acid, usually administered as the sodium salt. Bioavailability is dependent upon particle size and problems of generic inequivalence have therefore arisen, particularly in Scandinavia. The drug has a moderately large volume of distribution and is approximately 90% bound to plasma proteins. Clinically important displacement can be caused by bilirubin and several drugs, particularly sodium valproate, which is often combined with phenytoin. Displacement will lower the total serum concentration but will little affect the free drug concentration. The metabolism of phenytoin to the major metabolite, 5-(p-hydroxyphenyl)-5-(phenylhydantoin, is saturable, giving rise to a non linear dose-serum concentration relationship. Therefore, the dose range compatible with a therapeutic serum concentration is narrow within subjects, and monitoring serum concentrations is of particular value in dosage tailoring. In renal failure, the binding of phenytoin to plasma proteins is reduced and therefore a lower range of serum drug concentrations is compatible with therapeutic control. In liver disease, binding may also be impaired but delayed metabolism may occur in addition. During pregnancy the serum concentration may fall progressively as pregnancy advances, probably due to an increased rate of metabolism. Phenytoin readily crosses the placenta, and is metabolised rapidly by the neonate exposed in utero.


Transcutaneous oxygen measurement to evaluate drug effects. The evaluation of pharmacologic effects in perinatal medicine is dependent on our clinical methods for measurement and monitoring of the mother, the fetus, and the newborn infant. The development of a noninvasive continuous method of measuring PO2, the transcutaneous PO2 electrode, has greatly enhanced the ability to assess effects of drugs on the cardiorespiratory system. During labor, diazepam and meperidine have been documented to cause respiratory depression and significant decreases in PO2. The advantageous effect of epidural anesthesia on the oxygen-cardiorespirogram of mothers in labor has also been demonstrated. Both fetal and maternal tcPO2 have been successfully assessed during the administration of peridural catheter anesthesia (carticaine) and during suppression of labor with fenoterol. In the newborn infant, monitoring of tcPO2 has been helpful in assessing the residual effects of drugs administered during labor and delivery, in prescribing the appropriate and safe dose of oxygen, and in defining the effects of theophylline on the oxygen-cardiorespirogram. We have also demonstrated the effect of furosemide on PO2 in the treatment of pulmonary edema accompanying bronchopulmonary dysplasia and of indomethacin for the management of patent ductus arteriosus. Use of tcPO2 measurements for clinical pharmacologic evaluation is a promising addition to our research techniques.


Noninvasive monitoring of intracranial pressure: a new approach for neonatal clinical pharmacology. Measurements taken with the pressure-activated fiberoptic sensor should provide better insight into the relative usefulness of various agents such as glycerol and corticosteroids in the reduction of intracranial pressure. It is also useful in monitoring pressure changes during asphyxia-hypoxia, convulsions, and anesthesia, and in demonstrating the effects of drugs such as furosemide on intracranial pressure.


Fluid and electrolyte balance in very low birth weight infants. Correct fluid and electrolyte concentrations are essential to reduce mortality in critically ill infants. The authors list considerations in calculating the necessary maintenance volumes and emphasize the importance of continuous monitoring of data and a review and retabulation every eight hours, because requirements vary from one infant to the other, and in the same infant under different circumstances.


Chloramphenicol and the neonate--1979 view. In a therapeutic tragedy perhaps even more widespread than the thalidomide disaster, untold lives were lost between 1949 and 1958 through the administration of inappropriate doses of chloramphenicol to newborn infants. Sensitive assays of blood levels of chloramphenicol now available make it possible to employ this useful antibiotic avoiding its toxic effects. The author reviews accumulated knowledge about chloramphenicol in the newborn infant.


In utero opiate dependence and sudden infant death syndrome. Infants born to smoking, alcoholic, and drug-dependent women appear to suffer increased prenatal and postnatal growth deficiency, vascular changes, and hypoxic episodes as a result of decreased oxygen carrying capacity in the blood, depression with overdose, and withdrawal symptoms. The author reviews these associations and suggests further research to clarify their role in the sudden infant death syndrome.


Corticosteroid pharmacokinetics in liver disease. Among the corticosteroids, prednisone is the most commonly used in the treatment of chronic active liver disease. However, its pharmacokinetics have only recently been investigated. Prednisone is effectively absorbed and converted to its active therapeutic derivative, prednisolone, in healthy volunteers and in patients with liver disease; the bioavailability of oral prednisone approximates 100% of an intravenous dose and is comparable after administration of either prednisone or prednisolone. Patients with liver disease and hypoalbuminaemia are more likely to suffer major side effects of prednisone as a consequence of decreased protein binding and delayed clearance of prednisolone. Dosage in such patients should be reduced in accordance with serum albumin concentration.


Influence of drug exposure of the father on perinatal outcome. Initial skepticism that paternally administered drugs could damage the developing fetus has given way to concern that lead, narcotics, ethanol, anticonvulsants, anesthetic gases, caffeine, or cigarette smoke ingested at the time of conception might damage sperm or sperm motility or have indirect effects resulting in an increase in neonatal morbidity and mortality.


Gilbert's syndrome and drug metabolism.

Gilbert's syndrome and drug metabolism. Gilbert's syndrome is an inherited disorder which is characterised by unconjugated hyperbilirubinaemia. In patients with Gilbert's syndrome, both bilirubin clearance and in vitro hepatic microsomal uridine diphosphoglucuronyl transferase (UDPGT) activity are reduced. In addition, there is evidence suggesting impaired hepatic uptake of bilirubin in Gilbert's syndrome. Glucuronidation of a number of substrates appears to be impaired in Gilbert's syndrome, but the significance of the reported changes in oxidation and acetylation are less clear.


Use of serum creatinine concentrations to determine renal function. Serum creatinine concentrations are widely used clinically as an index of renal function. In stable normal or reduced renal function they are determined by the rate of creatinine production and the endogenous creatinine clearance, and, during changing renal function, also by the apparent volume of distribution of creatinine. These determinants of serum creatinine concentrations, however, are affected by age, sex and body weight. The rate of creatinine production is proportional to body weight, and it decreases with age and is slower in females than in males. The endogenous creatinine clearance decreases with age and is lower in females than in males. The apparent volume of distribution of creatinine is equal to the total body water, which is proportional to body weight, and it decreases with age and is lower in females than in males. The individual relationships between the determinants of serum creatinine concentrations and age, sex, and body weight on the relationship between normal and reduced renal function and serum creatinine concentrations are illustrated by simulations. Equations are derived to predict endogenous creatinine clearance from serum creatinine concentrations, and a nomogram is presented for determining relative renal function from serum creatinine concentrations, which take into account the age, sex and body weight of the individual patient. It is recommended for rapid clinical evaluation of stable normal or reduced renal function. When rapid changes are expected clinically, however, it is recommended to use a mid urine collection period serum creatinine sample for creatinine clearance determination.


Effect of drugs on intrauterine growth.

Effect of drugs on intrauterine growth. Although the teratogenic potential of maternally administered drugs is well known, their stimulation of intrauterine growth retardation may be equally deleterious. Possible adverse effects of propranolol, steroids, anticonvulsants, tranquilizers, and maternal smoking and drinking are discussed. Questions of fetal development and maternal management are considered in light of their interference with the mechanism of growth regulation.