The relationship between coding sequences and function in some heme binding proteins

Summary It is known that globin genes contain three exons with the middle exon coding for a four-helical supersecondary structure responsible for heme binding. Since this portion of the globin peptide chain can be structurally superimposed onto the cytochromec and cytochromeb ₅ chains (Argos and Rossmann 1979), it can be inferred that the cytochromec gene will contain only one coding sequence while the cytochromeb ₅ gene will be composed of three exons as found in the globin gene.     

High-performance liquid chromatographic and gas—liquid chromatographic determination of diazepam and nordiazepam in plasma

A one-step method for extraction of diazepam, nordiazepam, and internal standard into toluene is followed by chromatographic separation and detection with either dual-wavelength high-performance liquid chromatography or electron-capture gas—liquid chromatography. Agreement between the two methods was excellent for diazepam (r = 0.99, n = 38) and good for nordiazepam (r = 0.96, n = 79) over a concentration range that included subtherapeutic, therapeutic, and toxic plasma levels.     

Solubilization of rat heart sarcolemma 5′-nucleotidase by phosphatidylinositol-specific phospholipase C

The influence of a phosphatidylinositol-specific phospholipase C treatment on rat heart sarcolemmal 5′-nucleotidase was investigated. Upon complete hydrolysis of all phosphatidylinositol in the sarcolemma, 75% of 5′-nucleotidase activity was found in the solubilized form. The insolubilized enzyme after this treatment has the same K_m for AMP as the untreated, sarcolemmal-bound enzyme (0.04 mM), whereas the solubilized enzyme has a 40-fold increase in K_m for AMP (0.16 mM). Other sarcolemmal-bound enzymes were not affected by the same treatment. Hence, the specific involvement of phosphatidylinositol in the binding of 5′-nucleotidase to the sarcolemma of the rat heart is clearly demonstrated.     

2,4,6-Trinitrobenzenesulfonate labels an essential amino group near the bound phosphate at the catalytic site of mitochondrial F1-ATPase

The amino reagent 2,4,6-trinitrobenzenesulfonate (TNBS) was found to inactivate mitochondrial F₁-ATPase through covalent labeling, which was not reversed by dithiothreitol. The observed rate of inactivation was retarded by inorganic phosphate, but enhanced by prior labeling of F₁ with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-C1). These observations are consistent with the presence of an essential amino group near the bound inorganic phosphate at the catalytic site of F₁. A comparison of the observed protection of F₁ from NBD-C1 and 5′-p-fluorosulfonyl-benzoyladenosine (FSBA) respectively by inorganic phosphate and by 2′,3′-O-(2,4,6-trinitrophenyl)adenosine 5′-monophosphate (TNP-AMP) suggests that NBD-C1 labels an essential Tyr residue in the positively charged locus for binding the polyphosphate end of ATP, and that FSBA labels an essential Tyr residue in the more hydrophobic locus for binding the adenosine moiety of ATP at the catalytic site of F₁.     

Studies of the mechanisms for the induction of in vivo tumor immunity. V. A comparison of the generation of the primary cell-mediated cytotoxic response using in vitro mixed-lymphocyte--tumor cell culture and an in vivo technique

In vivo immunization of C57BL/6 mice with FBL-3, a syngeneic Friend-virus-induced leukemia, can induce a specific, T-cell-mediated cytotoxic response, as measured by the ¹²⁵IUdR release assay. In vitro it was difficult to generate an analogous, primary cytotoxic response, using a normal spleen responder population. After modification of the splenic responders by adding normal peritoneal cells, this modified population then had the capacity to mount a primary cytotoxic response in the mixed-lymphocyte-tumor cell culture reaction to FBL-3. We have characterized the effector population as well as the helper (peritoneal) cells which were responsible for elevating the cytolytic response to FBL-3. The results indicate that there are at least two populations of cells which are essential for inducing a primary cell-mediated cytotoxic response. First, the effectors which are directly responsible for mediating the cytotoxic reactions and are derived from radiosensitive T cells and second, a helper cell population which is radioresistant and has the characteristics of macrophages.     

Na movements and their oscillations during fertilization and the cell cycle in sea urchin eggs

We have analysed in detail the Na⁺ content and Na⁺ influx during fertilization and first divisions of the sea urchin egg (Paracentrotus lividus) using a filtration technique devised to eliminate rapidly contamination by the Na⁺ of external sea water. In the first 5 min following fertilization the egg fills up with Na⁺ (+ 30%). Thereafter Na⁺ is extruded and the Na⁺ content stabilizes at about 60% of the unfertilized egg level by the second cleavage (2 h). The initial increase in Na⁺ content is due to a large increase in Na⁺ influx already detected at 20 sec. The Na⁺ influx reaches its maximum at 1 min and its minimum at 5 min. H⁺ excretion follows the same kinetics. A second increase in Na⁺ influx is noted 5–10 min after fertilization; it reaches its maximum at prophase metaphase (30 min) and its minimum during cleavage (60 min). These oscillations in Na⁺ influx were observed for the first three divisions. Fertilization also immediately stimulates the Na⁺ efflux which remains elevated throughout the cell cycle and is responsible for the depletion of the Na⁺ content of the embryos. Activation of the eggs by weak amine bases (5 mM NH₄Cl) which bypasses the early cortical reaction produces only a depletion in the Na⁺ content of the egg similar to that produced by fertilization. NH₄Cl also increases the Na⁺ influx soon after fertilization, although no transient variations are noted.     

Les Radiolaires d'âge jurassique supérieurà crétacé supérieur dans les Radiolarites du Pinde-Olonos (Presqu'île de Koroni; Péloponnèse méridional,Grèce)

Radiolarians of several radiolarites sections of the Pindos-Olonos zone, southern Peloponnesus, lead to propose direct datations of mesozoic sediments. The acuity of such datations allow to distinguish two periods for depositionof radiolarites s.s. in the tethyan region. The first period would be Upper Jurassic and general in tethyan realm, radiolarites depositing under various latitudes (0–35°N). This would be the result of a strong ocean surface current. The second period would be Upper Cretaceous (Vraconian—Coniacian) and be geographically much more restricted (0–15°N). This could result from a weaker current. The absence of radiolaritic sedimentation is possibly the result of the destruction or insulation of the accurate basins during the obduction of ophiolites on the apulo-african realm. The inventory of Late Cretaceous radiolarian fauna, rarely done on alpine series so far, shows similar result to those obtained in Central American and California.     

Une nouvelle sous-famille,les Poulpinae, et quatre nouvelles espèces de Saitoum radiolaires mésozoiques téthysiens

Studies of skeleton elements of several jurassic species of Saitoum allow to compare them with Poulpus from Trias. The sub-family Poulpinae is introduced, caracterised by three cephalic arcs and the collar position of the cephalic structure. Among the jurassic forms, 4 species are newly described: S. corniculum, S. elegans, S. levium and S. trichylum.     

Oxygen availability,redox and the distribution of some Turbellaria Schizorhynchidae and other forms

Records from a considerable number of meiofaunal samples are reexamined in relation to the depth within sediment distribution of various taxa especially Schizorhynchidae and the main turbellarian orders.Frequency and density records in samples with particular oxygen flow rates and redox potentials confirm that the ranges of Schizochilus, Proschizorhynchus and Neoschizorhynchus spp. within these factors may help account for congeneric occurrence within cores. However, statistical analysis of the records is seldom able to confirm apparent differences. There is, for example, a statistically significant difference in distribution in relation to redox potential between the red schizorhynchians Pseudoschizorhynchides ruber, Diascorhynchus rubrus and the red acoelan Paratomella rubra but not in relation to oxygen availability and depth within sediment. On one beach Gnathostomula aff. paradoxa was found at Eh and O₂ availability values under which it must be assumed to be living anaerobically.Contingency and other analyses are extended to differences between higher taxa and the results discussed in relation to habitat evolution of the Turbellaria. The suggestion that structural complexity of turbellarians has increased with increased oxygenation of the environment is tentatively supported.