Radioimmunoassay for the detection and quantitation of bruceantin

A radioimmunoassay for a new anticancer drug, bruceantin, has been developed using [³H]acetylbruceantin and antibody induced by immunizing rabbits with succinylbruceantin-bovine serum albumin conjugates. [³H]Acetylbruceantin was synthesized by reacting bruceantin with [³H]acetyl anhydride. The assay is simple and reproducible. The standard curve was linear on a logit-log plot, and the lower limit of sensitivity of the assay was 1 ng/ml. Using this assay, drug levels were easily determined in tissues of experimental animals following bruceantin administration. The assay procedure does not require sample extraction for plasma, urine, and bile. Bruceantin in other tissues can be extracted quantitatively with ethanol before being measured by the radioimmunoassay.     

Trisomy 6p22→6pter due to familial t(6;13)(p22;q34 or 33) translocation

Summary 235 cases of Down's syndrome were ascertained in a 10-year study of Down's syndrome in Western Australia. Although cytogenetic studies performed on 222 subjects confirmed that 95% of cases were trisomic due to nondisjunction, 4% were trisomic due to translocation, and 1% were mosaic, the ratio of inherited/sporadic translocations differed from that usually reported. Comparison of the results with those of an earlier Australian survey of Down's syndrome demonstrated a real fall in the incidence of Down's syndrome in Australia but no significant change in maternal age-specific incidences.     

Kälteresistenz der Fichte

Fichtenchloroplasten durchlaufen während eines Jahres saisonbedingte charakteristische Struktur- und Funktionswechsel: Frühjahrschloroplasten, die in den alten Nadeln kurz vor und während des Knospenaustriebes gefunden werden, sind sehr groß und so sehr mit Stärke erfüllt, daß man sie als Amyloplasten bezeichnen kann. Beim Aufbau der neuen Nadelgeneration wird diese Stärke verbraucht, und es entwickeln sich aus den Amyloplasten wieder die photosynthetisch aktiven Sommerpiastiden mit einem gut entwickelten Membransystem. Während der Frosthärtung und insbesondere während der Frostperiode treten die Chloroplasten in ihrer Winterform auf: amöboide, an einer Stelle der Zelle konzentrierte, stark aufgequollene Plastiden mit einem aufgelockerten und zum Teil reduzierten Thylakoidsystem. Der Strukturwandel der Chloroplasten wird — wie die Frostresistenz selbst — durch exogene Faktoren (Tageslänge, Temperatur) ausgelöst und kann auch durch artifiziell veränderte Umgebung zu unnatürlichen Zeitpunkten ausgelöst werden. Begleitet wird der Strukturwandel von einer Veränderung der CO₂-Fixierungsrate der Fichtennadeln, die ihrerseits auf Veränderungen der photochemischen Aktivität der Chloroplasten zurückzuführen ist. In Frostexperimenten konnte gezeigt werden, daß die Frosthärtung Reaktionen auf zwei verschiedenen Ebenen auslöst: 1. Produktion von kolligativ wirksamen Membranschutzstoffen sowie 2. einer Veränderung der chemischen Zusammensetzung der Chloroplastenmembranen; zumindest letztere führt offensichtlich zu einer Verringerung der Photosyntheseleistung. Frostschädigung der Chloroplasten tritt bei nicht entsprechend stark gehärteten Nadeln auf, jedoch nicht durch direkten Einfluß der tiefen Temperatur auf das Thylakoidsystem als vielmehr durch Freisetzung membranschädigender Substanzen durch Permeabilitätsverlust plasmatischer Membranen. Wir danken Herrn Prof. Dr. O. Kandler für anregende Diskussionen und für die kritische Durchsicht des Manuskriptes. Der Deutschen Forschungsgemeinschaft wird für die finanzielle Unterstützung dieser Arbeit gedankt.     

Improved basal medium for Y-1 mouse adrenal cortex tumor cells in culture

Summary An improved basal medium is presented that requires only minimal supplementation with dialyzed fetal bovine serum or bovine serum albumin and fetuin to be comparable to Ham's F-10, which requires 15% horse serum (HS) and 2.5% fetal bovine serum (FBS) for the growth and function of Y-1, mouse adrenal cortex tumor, cells. Cell monolayers maintained for up to 2 weeks without any protein supplementation have retained their steroid response to ACTH. The medium differs from Ham's F-10 in its buffer composition and higher calcium-ion concentration. This medium should be a useful adjunct to studies pertaining to steroid and lipid intermediary metabolism, the retention of a specialized physiological function in a chemically defined medium, and the mechanism of hormonal response. Supported by the Medical Research Service of the Veterans Administration.     

Biochemical and genetic analysis of a mutant with altered alkaline phosphatase activity in Dictyostelium discoideum

Alkaline phosphatase is one of several enzymes that accumulate in a temporally regulated sequence during the development of Dictyostelium discoideum. These enzymes can be used to monitor specific gene expression; moreover, isolation and analysis of mutations in the structural gene(s) can serve to indicate some of the essential steps in programmed synthesis and morphogenesis. A mutation (alpA) which affects the activity and substrate affinity of alkaline phosphatase was isolated in D discoideum using a procedure for screening large numbers of clones. Alkaline phosphatase activity at all stages of vegetative growth and development was altered by the mutation. Several physical properties of the enzyme from growing cells and developed cells were compared and found to be indistinguishable. It is likely that a single enzyme is responsible for the majority of alkaline phosphatase activity in growth and development. The mutation is coexpressed in diploids heterozygous for alpA and maps to linkage group III. One of the haploid segregants isolated from these diploids carries convenient markers on each of the six defined linkage groups and can be used for linkage analysis of other genetic loci.     

Growth of “Seeded” Cellulolytic Enrichment Cultures on Mesquite Wood

Eleven enrichment cultures were developed by a “seeded” enrichment culture technique, and one was developed by a simple enrichment technique. The seeded enrichment, the pure “seed,” and the simple enrichment cultures were compared during growth on mesquite wood, cotton, carboxymethylcellulose, and cellobiose. All of the enrichment cultures were cellulolytic and exceeded the pure seed cultures in mesquite wood hydrolysis and/or viable cell count. Yeast extract improved, but was not essential for, growth of the seeded enrichment cultures on carboxymethylcellulose. Two of the seeded enrichment cultures, CAD5 and CAD11, grew best at 37°C and pH 7.0 on mesquite wood. A 1.0% (wt/vol) wood concentration was optimum for their growth.     

Pyruvate metabolism after in vivo exposure to oral arsenic

This study investigated altered pyruvate metabolism after prolonged oral arsenic exposure. Male rats were given access to deionized drinking water containing 0, 40 or 85 ppm sodium arsenate (As⁵⁺) for 3 weeks. Respiration studies with mitochondria isolated from treated animals indicated decreased state 3 respiration (with ADP) and decreased respiratory control ratios (RCR) for pyruvate/malate-mediated respiration, but not for succinate-mediated respiration, as compared to control respiration values. In addition, pyruvate dehydrogenase activity was measured, in both liver and intestine, before and after Mg-activation in vitro. After 3 weeks, the effects of arsenic at the highest dose level were pronounced on the basal pyruvate dehydrogenase activity (before activation) as well as the total pyruvate dehydrogenase (after activation). The inhibition of pyruvate dehydrogenase activity both before and after Mg-activation suggests an arsenic effect on mitochondrial pyruvate metabolism which, in part, involves inhibition of pyruvate decarboxylase. Evidence is also presented which may indicate an arsenic effect on the kinase and/or phosphatase which regulate pyruvate dehydrogenase activity.     

The immune response of allophenic mice to the synthetic polymer GLΦ

The immune response of allophenic mice of type C57BL/6(A × SJL) F₁ to GL administered in complete Freund's adjuvant was tested. Control mice of the three strains C57BL/6, A, and SJL are all nonresponders to this antigen. However, the F₁ generations of C57BL/6 × A, C57BL/6 × SJL, and A × SJL were all responders to the antigen, so that the complementarity of at least two genes is confirmed. The allophenic mice showed no further complementation beyond the F₁ generation, a result which may argue against the possibility that more than two genes control the response to GL in these mouse strains. Characterization of the allophenic mice over several months showed that they exhibit chimeric drift, both in their coat color and in peripheral white blood cell population. There is no apparent correlation of coat color to the lymphocyte composition of the mice at any one time. The mice are true chimeras, since killing of the two populations of white blood cells with two different anti-H-2 sera produced a 100 percent killing. The immune response of individual allophenic mice to GL showed a good correlation to the number of A × SJL lympho-cytes in the animal.Abbreviations used in this paper are GL an amino acid polymer of 57 %l-glutamic acid, 38%l-lysine, and 5%l-phenylalanine - GLT¹⁵ an amino acid polymer ofl-glutamic acid,l-lysine, and 15 %l-tyrosine - (T,G)-A-L an amino acid polymer having a polylysine backbone with side chains of polyd-l-alanine, terminating in short sequences of tyrosine and glutamic acid - GAT₁₀ an amino acid polymer of 60%l-glutamic acid, 30%l-alanine, and 10%l-tyrosine - GLA₅ an amino acid polymer of 57%l-glutamic acid, 38%l-lysine, and 5%l-alanine - DNP 2,4 dinitrophenyl - BGG bovine gamma globulin - FCS fetal calf serum - PWBC peripheral white blood cell - SWBC spleen white blood cell - T cell thymus-derived lymphocyte - B cell bone marrow-derived lymphocyte     

The Histochemical Localization of Cholesterol in Formalin-Fixed and Fresh Frozen Sections

The premixed cholesterol reagent (acetic acid, sulphuric acid, ferric chloride and phosphoric acid) employed in a spectrophotometric method for serum cholesterol determinations was applied to frozen sections of various rat and rabbit tissues. The histochemical reaction was compared with the results obtained by the classical Schultz reaction. The same bluish-green color was observed after treatment of formalin-fixed and fresh cryostat sections incubated in 2.5% iron alum at 37 C for 3 days. The premixed cholesterol reagent is stable for years at room temperature if stored in a brown bottle and is thus readily available for routine use.