The peripheral-type benzodiazepine receptor. Localization to the mitochondrial outer membrane

We have investigated the subcellular localization of the peripheral-type benzodiazepine receptor in rat adrenal gland using the high affinity ligand 3H-labeled 1-(2-chlorophenyl)-N-methyl-(1-methylpropyl)-3-isoquinoline carboxamide ([3H]PK11195). The autoradiographic pattern of [3H]PK11195 binding sites in tissue sections of adrenal gland is similar to the histochemical distribution of the mitochondrial marker enzymes, cytochrome oxidase and monoamine oxidase, which are present in high concentrations only in the cortex. Subcellular fractionation studies of homogenates of adrenal gland indicate that the recovery and enrichment of [3H]PK11195 binding sites in the nuclear, mitochondrial, microsomal, and soluble fractions correlate closely with cytochrome oxidase activity, but not with markers for the nuclei, lysosomes, peroxysomes, endoplasmic reticulum, plasma membrane, or cytoplasm, indicating an association of the peripheral-type benzodiazepine receptor with the mitochondrial compartment. Titration of isolated mitochondria with digitonin results in the simultaneous release of the peripheral-type benzodiazepine receptor and of monoamine oxidase, but not cytochrome oxidase, indicating association of the peripheral-type benzodiazepine receptor with the mitochondrial outer membrane. Scatchard analysis and drug displacement studies of the binding of [3H] PK11195 to intact mitochondria and to the outer membrane-enriched digitonin extract further confirm the localization of the peripheral-type benzodiazepine receptor to the mitochondrial outer membrane.     

Isolation and reconstitution of an N-ethylmaleimide-sensitive phosphate transport protein from rat liver mitochondria

An N-ethylmaleimide-sensitive phosphate transport protein has been isolated from rat liver mitochondria, substantially purified, and reconstituted into phospholipid vesicles. Purified inner mitochondrial membrane vesicles depleted of F1-ATPase by urea treatment proved to be the most satisfactory starting material. Treatment of these membrane vesicles with Triton X-100 resulted in solubilization of the phosphate transport protein. Further purification was achieved using hydroxylapatite powder. Polyacrylamide gel electrophoresis of the purified fraction in sodium dodecyl sulfate indicated the presence of two Coomassie blue-staining bands with apparent Mr's of 30,000 and 35,000. Labeling of the 35,000 Mr band by the Pi transport inhibitor diazobenzene sulfonate was reduced markedly by prior treatment of the mitochondria with the inhibitor N-ethylmaleimide. The purified fraction containing both proteins could be reconstituted into liposomes prepared from purified asolectin. Phosphate efflux from these vesicles was inhibited by N-ethylmaleimide, by the impermeant mercurial agent, p-chloromercuribenzoate, and by diazobenzene sulfonate. Treatment of the purified fraction with N-ethylmaleimide prior to incorporation into liposomes resulted in a reconstituted system incapable of catalyzing Pi efflux. These studies summarize the first detailed attempt to purify the Pi/H+ transport system from rat liver mitochondria and emphasize the need to commence the purification with purified inner membrane vesicles depleted of F1-ATPase. In addition, these studies show that the final fraction contains a reconstitutively active transport system which when incorporated into phospholipid vesicles has its essential sulfhydryl groups oriented outward. Finally, it is shown that the purified fraction also contains a 30,000 Mr component.     

Activated sludge treatment of synthetic wastewater containing pentachlorophenol

Activated sludge treatment of a pentachlorophenol (PCP)-containing synthetic waste was examined. With a waste containing some sugars, and 40-120 mg/L PCP, laboratory activated sludge required about seven days for acclimation. However, the prior addition of a quasipure culture of PCP-metabolizing Arthrobacter resulted in immediate acclimation. Even with acclimated sludge, however, the system was upset for two days by a simple step change from 40 to 120 mg/L of PCP. The stability of the system to such a shock load was considerably improved when a chemostat culture of the PCP-metabolizing Arthrobacter was fed slowly into the mixed liquor. Kinetic models were developed to describe the dynamic response of the system in terms of growth parameters, hydraulic detention time, sludge age, and bleed-in rate from the chemostat.     

Afternoon plasma cortisol in depressed patients: A measure of diagnosis or severity?

Plasma cortisol concentration was measured at 20 min intervals from 3 p.m. (1500 hrs) to 6 p.m. (1800 hrs) in 26 hospitalized patients classified according to the Newcastle Index as endogenously depressed (n = 16) or non-endogenously depressed (n = 10). When examined in depressed state, before treatment, maximum, mean and range of plasma cortisol concentration in this time interval was significantly higher in the endogenously depressed patients than in the non-endogenously depressed patients (p less than 0.01-0.02). The diagnostic identification of endogenous depression on the basis of these cortisol concentration measurements was at least as good as that reported by others using post-dexamethasone cortisol levels. The plasma cortisol levels (maximum, mean) and fluctuations (range) correlated significantly with the degree of depression (Hamilton Depression Scale), and differences in severity of depression could explain most of the differences in cortisol levels between the two diagnostic groups. Nine patients were reexamined after 3-12 months in a non-depressed state, and all unipolar endogenously depressed patients (n = 6) then had clearly reduced cortisol levels and fluctuations.     

The 11q;22q translocation: A European collaborative analysis of 43 cases

Summary Translocation between the long arms of chromosomes 11 and 22 is usually detected in offspring with an unbalanced karyotype following a 3:1 disjunction resulting in partial trisomy. Since by the end of 1976 it was suspected that this translocation might be more frequent than one would deduce from published reports, it was decided to call for a collaborative effort in Europe to collect unpublished cases. In response, 42 cases were collected in Europe, and one case from New Zealand was added. The following countries were represented with the number of cases indicated in parentheses: Czechoslovakia (2), Denmark (4), Finland (3), France (6), Germany (1), Italy (5), The Netherlands (9), Sweden (6), United Kingdom (4), Yugoslavia (2). The wide geographical distribution indicates a multifocal origin of the translocation. Among the unpublished cases, 31 were ascertained as unbalanced carriers [47,XX or XY,+der(22),t(11;22)] and 12 as balanced balanced carriers [46,XX and XY,t(11;22)]. Among the published cases, 10 were ascertained in unbalanced and 3 in balanced carriers. The breakpoints of the translocations indicated by the contributors varied, the most frequently reported being 11q23;22q11 (25 cases), followed by q25;q13 (10 cases). While the first one seems more likely, it was not possible to decide whether the breakpoints were the same in all cases.All 32 probands with unbalanced karyotypes had inherited the translocation, 31 from the mother and only 1 from the father. This ratio became 43:1 when the published cases were added. A segregation analysis revealed that in families ascertained through probands with unbalanced karyotypes there was a ratio of carriers to normal (all karyotyped) 54:55, not a significant difference. The formal maximum (minimum) recurrence risk for this unbalanced translocation was calculated to be 5.6% (2.7%). When the ascertainment was through a balanced proband, the maximum risk was 2.7%. The risk was calculated as 5.7% for female and 4.3% for male carriers. The mean family size was 1.67 for the offspring of female carriers and 0.78 for the offspring of male carriers. This significant difference suggests that heterozygosity for the translocation reduces fertility in males. Indeed, several of the probands with balanced karyotypes were ascertained because of sub- or infertility. Only 2 de novo translocations were found among the 59 probands, and both, were among the 12 cases ascertained as balanced carriers. The source, quality, and quantity of the clinical data for the subjects with unbalanced karyotypes were variable, and no definite conclusions were possible about phenotypes. The following signs were recorded in 10 or more of the 45 cases: low birth weight, delayed psychomotor development, hypotonia, microcephaly, craniofacial asymmetry, malformed ears with pits and tags, cleft palate, micro-/retrognathia, large beaked nose, strabismus, congenital heart disease, cryptorchidism, and congenital dislocation of the hip joints. Many signs were similar to those considered typical of trisomy 11q, and the phenotype coincided almost completely with the presumptive phenotype of complete trisomy 22. No cases with coloboma was recorded, while other signs of the cat-eye syndrome were found in several probands. This might indicate that individuals with the cat-eye syndrome and carriers of the unbalanced 11/22 translocation have the same segment of 22 in triplicate plus or minus another chromosome segment.     

Culture studies on the rare brown alga Phaeostroma longisetum comb.nov. and its common relative P. pustulosum from Greenland

Culture studies have shown that the rarely reported alga, Symphyocarpus longisetus , agrees with the type species of Phaeostroma with regard to its mode of formation of plurilocular sporangia, number of chloroplasts, and type of true hairs, and disagrees with the type species of Symphyocarpus. The new combination, Phaeostroma longisetum , is proposed. Unilocular sporangia are reported for the first time in this species. The life history is of the direct type under all conditions tested. The species grows and reproduces also at 15°C LD conditions although generally considered an arctic species.
Phaeostroma pustulosum probably also has a homophasic, direct type of life history. The plants are either pseudoparenchymatous discs or branched filaments. The filamentous thalli are in agreement with the younger taxon Phaeostroma aequale (= Streblonema aequale).     

A hybridization assay specific for ribosomal RNA from Escherichia coli

Summary Ribosomal RNA fromEscherichia coli forms specific hybrids with heterologous DNA fromVibrio metschnicovii. The conditions for such hybridization are described, in which the amount of mRNA fromE. coli binding to theVibrio DNA is about 3% of the amount which hybridizes to the same amount of DNA fromE. coli.     

The “azotopure” process for treating nitrogen-deficient aqueous wastes

Many chemical and food processing wastes are deficient in nitrogen, and must therefore be supplemented with domestic sewage or ammonia to dispose of them by conventional biological treatment. A more economical treatment, especially of strong wastes, is possible by maintaining a semipure culture of nitrogen-fixing bacteria in a stirred aerated tank. When operated as a chemostat with 4 to 6 hr detention time, removal of 85% of the COD is possible in a single step without the need for clarifiers or digesters. A clarified effluent could also be produced by flotation or sedimentation of the flocculated bacteria. In that case there would be 99% removal of COD and a reduction in the amount of sludge by two-thirds over the conventional activated sludge process.     

Different Classes of Ribonucleic Acid Isolated from Lymphocytic Choriomeningitis Virus

Purified preparations of lymphocytic choriomeningitis virus (LCM virus) contain three classes of RNA. The previously described 18s, 23s, 28s, and 31s RNAs, where the 23s and 31s RNAs are viral-specific, and the 18s and 28s RNAs probably are host RNAs incorporated in the virion. Now, 4s, 5s, and 5.5s RNAs can be isolated as well. Thus five RNAs which migrate by acrylamide gel electrophoresis as ribosome-derived RNA can be isolated from purified LCM virus. This observation further supports the reports that arenaviruses may contain ribosomes. The ribosome-derived RNA can be synthesized both before and after the virus infection. The viral 23s could be a hydrogen-bonded complex forming the 31s RNA, or it could be contained in defective interfering LCM virus particles; these possibilities are examined.