Rapid purification and direct spectrophotometric assay for 5-aminolevulinic acid dehydratase

A two-step purification procedure for 5-aminolevulinic acid dehydratase (EC 4.2.1.24) from human red blood cells has been developed. It involves one ion exchange and one gel filtration step. The purification is about 1000-fold, and the yield is more than 85%. With the purified enzyme a direct spectrophotometric assay of product formation without subsequent reaction with Ehrlich's reagent is described.     

Purification and properties of NADPH-dependent aldehyde reductase from human liver.

An aldehyde reductase (EC 1.1.1.2) from human liver has been purified to homogeneity. The enzyme is NADPH-dependent, prefers aromatic to aliphatic aldehydes as substrates, and is inhibited by barbiturates and hydantoins. The following physicochemical parameters were determined: molecular weight, 36,200; sedimentation coefficient, 2.9 S; Stokes radius, 2.65 nm; isoelectric point, pH 5.3; extinction coefficient at 280 nm, 54,300 M-1 cm-1. Results from polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate, gel filtration, and ultracentrifugation suggest a monomeric structure. On molecule of NADPH binds to the enzyme causing a red shift of the coenzyme absorption maximum from 340 to 352 nm. The amino acid composition has been determined and a partial specific volume of 0.74 was computed from these data. An alpha-helicity of 7 and 18% was estimated from the ellipticities at 208 and 222 nm, respectively. Combination of the most reactive thiol group with p-mercuribenzoate does not cause loss of catalytic activity. Inactivation occurs when more than one thiol group is modified. The presence of NADPH or NADP+ prevents loss of activity by thiol modification. The comparison of structural features of aldehyde reductase with other monomeric and oligomeric dehydrogenases suggest similarities of aldehyde reductase with octopine dehydrogenase.     

Tauchbeobachtungen an Plankton und an Echostreuschichten

Zusammenfassung 1. Das Tauchen als Methode zur Untersuchung von Plankton und Echostreuschichten wird durch vier Beispiele erläutert: (a) Visuelle Beobachtungen an Wasserschichtungen und Grenzschichten durch Schwimmtaucher. (b) Untersuchung von Echostreuschichten durch Freitaucher, wobei sich ergab, daß angesammeltes biogenes Material in den untersuchten Sprung- beziehungsweise Streuschichten die Schallreflektion nicht beeinflußt. (c) Beobachtung von Großplankton und Feststellung von Planktonund Sestonkonzentrationen beim Tauchen mit dem Bathyscaph. (d) Untersuchung der Tiefenstreuschicht (deep scattering layer) durch Beobachtung der Vertikalwanderung bestimmter Arten des Großplanktons mit den Tauchbooten Bathyscaph und Soucoupe Plongeante. Physonectide Siphonophoren und Myctophiden standen in deutlicher Beziehung zur Tiefenstreuschicht und wurden als Echogeber erkannt.2. Die Möglichkeiten, von Tauchbooten aus quantitative und qualitative Proben von Plankton und auch vom Benthos zu nehmen, sind zur Zeit noch unzureichend. Die Entwicklung entsprechender Geräte für den wahlweisen und mehrfachen Einsatz bei demselben Tauchgang wird empfohlen.

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Diving observations on plankton and on scattering layers

Diving techniques are employed as a research tool in plankton investigations carried out in shallow water of the western Baltic Sea. Observations and samplings were made by skin divers on scattering layers corresponding to the discontinuity layers. Biogene materials, sometimes concentrated at the thermocline, are not responsible for this special kind of scattering, but rather discontinuity of salinity and temperature (Lenz 1965). For observations in deep water the use of undersea vehicles is recommended. From the Bathyscaph and the diving saucer, single plankton organisms and plankton concentrations were observed (e. g.Bernard 1958); investigations on the deep scattering layer have shown physonectid siphonophores and myctophids to be scatterers (Barham 1966). The equipment for sampling plankton and benthos from undersea vehicles is poorly developed. We need urgently gear for quantitative and qualitative sampling and for manifold use during single dives, i. e., multiple sampling gear and magazins for storage of samples.

     

On the pharmacology of venous smooth muscle from dog and man.

Changes in tension of spiral strips from canine and human veins induced by various drugs are compared with results from the literature on human veins. The order of potencies of alpha-adrenoceptor stimulating amines (adrenaline greater than noradrenaline greater than dopamine greater than isoprenaline) is similar in human and canine veins. Comparison of the affinities for alpha-adrenoceptors of pharmacologic drugs (thymoxamine, indoramine, clonidine, dihydroergotamine) suggest marked differences between the alpha-adrenoceptors in veins from man and dog. Venoconstriction mediating 5-HT receptors and a very small population of beta-adrenoceptors exist in both species. Human veins are always dilated by histamine, while canine femoral veins in vitro are relaxed by lower and contracted by higher histamine concentrations. Prostaglandin F2 alpha constricts both canine femoral and human hand veins. PGA2 and PGE2 increase the tension of canine and human veins in vitro but dilate human hand veins in situ. The order of potencies of ergot alkaloids in canine femoral veins is ergotamine greater than dihydroergovaline greater than dihydroergotamine = dihydroergostine greater than methysergide, whereas in human hand veins it is ergotamine = dihydroergovaline greater than methysergide greater than dihydroergotamine greater than dihydroergostine. In dogs the venoconstrictor effect of ergotamine is mediated by at least 3 mechanisms: stimulation of [1] alpha-adrenoceptors, [2] 5-HT receptors and [3] endogenous prostaglandin synthesis. Stimulation of alpha-adrenoceptors by dihydroergotamine and of 5-HT receptors by ergotamine was confirmed on human hand veins in situ. Prejunctional receptors at sympathetic nerve terminals are involved in the regulation of venous tone. Inhibitory alpha-adrenoceptors, dopamine and PGE2 receptors as well as facilitating beta-adrenoceptor existing at human vasoconstrictor nerves may be stimulated or blocked by pharmacologic drugs thereby modifying venous tone.     

Regulation of thyroid hormone metabolism in rat liver fractions

The nature of the conversion of thyroxine (T₄) to triiodothyronine (T₃) and reverse triiodothyronine (rT₃) was investigated in rat liver homogenate and microsomes. A 6-fold rise of T₃ and 2.5-fold rise of rT₃ levels determined by specific radioimmunoassays was observed over 6 h after the addition of T₄. An enzymic process is suggested that converts T₄ to T₃ and rT₃. For T₃ the optimal pH is 6 and for rT₃, 9.5. The converting activity for both T₃ and rT₃ is temperature dependent and can be suppressed by heat, H₂O₂, merthiolate and by 5-propyl-2-thiouracil. rT₃ and to a lesser degree iodide, were able to inhibit the production of T₃ in a dose related fashion. Therefore the pH dependendy, rT₃ and iodide may regulate the availability of T₃ or rT₃ depending on the metabolic requirements of thyroid hormones.