(Na + K)-ATPase activity of crude homogenates of rat skeletal muscle as estimated from their K-dependent 3-O-methylfluorescein phosphatase activity

A highly sensitive fluorimetric assay using 3-O-methylfluorescein phosphate as substrate was used in the determination of K⁺-dependent phosphatase activity in preparations of rat skeletal muscle. The gastrocnemius muscle was chosen because of mixed fibre composition. Crude, detergent treated homogenate was used so as to avoid loss of activity during purification. K⁺-dependent phosphatase activities in the range 0.19–0.37 μmol · (g wet weight)⁻¹ · min⁻¹ were obtained, the value decreasing with age and K⁺-deficiency. Complete inhibition of the K⁺-dependent phosphatase was obtained with 10⁻³ M ouabain. Using a KSCN-extracted muscle enzyme the intimate relation between K⁺-dependent phosphatase activity and (Na⁺ + K⁺)-activated ATP hydrolysis could be demonstrated. A molecular activity of 620 min⁻¹ was estimated from simultaneous determination of K⁺-dependent phosphatase activity and [³H]ouabain binding capacity using the partially purified enzyme preparation. The corresponding enzyme concentration in the crude homogenates was calculated and corresponded well with the number of [³H]ouabain binding sites measured in intact muscles or biopsies hereof.     

Parturition and alate morph determination in the aphid Megoura viciae

In reproducing apterae of Megoura viciae, parturition is often completely arrested during periods of isolation from the host plant. In contrast, surgical removal of the rostrum (including the stylets), amputation of the extremities of the legs, or decapitation, all stimulate parturition away from the plant. These operations also induce alata-producing aphids to revert immediately to the production of apterae, but have no detectable effect on aptera-producers. Carbon dioxide or ether anaesthesia and nitrogen narcosis have a similar action on this maternally controlled response. Although the rostrum and tibio-tarsus bear sensilla whose removal might well be involved in inducing parturition, the influence on morph change is probably indirect and is to a great extent associated with the delay in the resumption of parturition. The effect can be reproduced by isolating individual aphids away from the food plant. The morph change cannot, however, be attributed to starvation since it also occurs when the genital pore of an actively feeding aphid is temporarily occluded. The change in physiology appears to be associated with the retention of embryos at a time when there is no sensory input from crowding.

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Résumé Chez les Megoura viciae aptères, la parturition est souvent complètement arrêtée pendant les périodes de séparation de la plante hôte. Par opposition, l'amputation chirurgicale du rostre (y compris les stylets), de l'extrémité des pattes (tarse et une partie du tibia), ou la décapitation, stimulent toutes la parturition en l'absence de la plante. Les pucerons induits à produire des ailés (élevage antérieur en groupe) retournent, après ces opérations, immédiatement vers la production d'aptères. Le dioxyde de carbone ou l'anesthésie à l'éther et la narcose à l'azote ont une action semblable sur ce déterminisme maternel. Bien que l'élimination des sensilla portées par le rostre et par l'ensemble tarse-tibia puisse être déterminante dans l'induction de la parturition, l'action sur le changement de type semble être principalement associée au retard consécutif dans la reprise de la parturition. Les anesthésiants qui, eux aussi, retardent l'apparition ou la reprise de la parturition, ont probablement une action indirecte du même type.Les pucerons groupés, isolés de la plante hôte pendant plus de 24 h, ont aussi tendance à retourner immédiatement à la production d'aptères. Ce changement de type ne peut, cependant, être attribué au jeûne puisqu'il se produit aussi quand, chez un puceron s'alimentant activement, le pore génital est momentanément bouché. Le changement physiologique semble être associé à la rétention des embryons à un moment où il n'y a pas l'influence sensorielle du groupement.Aucun de ces traitements, à l'exception du groupement, n'induit des pucerons antérieurement isolés à devenir des producteurs d'ailés.

     

A continuous-flow, rapid-mixing, photolabeling technique applied to the acetylcholine receptor

A continuous-flow technique is described in which a photoaffinity label, membrane rich in acetylcholine receptor, and various effectors are rapidly mixed, passed through a delay tube, through a tube in which they are irradiated, and are collected in a tube containing quencher. Delay times as short as 20 ms between mixing and photolysis are achievable. Because the flow is continuous, milliliter volumes of membrane can be labeled in a single run, which is convenient for the analysis of both the functional effects and sites of photolabeling. Using this technique, we have found that receptor in its transitory, active state, in which the channel is open, is more susceptible to photolabeling by the noncompetitive inhibitor analog [3H] quinacrine azide than is receptor in either its resting or desensitized states, in which the channel is closed. This technique should prove generally useful for the photolabeling of transient conformational states of macromolecules.     

Colorimetric determination of microgram quantities of polylysine by trypan blue precipitation

A simple and sensitive method for the determination of polylysine in solution is described. Polylysine is quantitatively precipitated with trypan blue. The absorbance of unbound dye in the supernatant is inversely proportional to the concentration of this polyamino acid. The precipitation is identical for all sizes of polylysine of molecular weight 13,000 or higher, and is prevented by the addition of either polyanions or serum. The measurable range of polylysine hydrobromide is between 1 and 10 micrograms/ml, which is about 10-fold lower than that by the published methyl orange precipitation method.     

A comparison of two methods for the microscopic determination of age at death.

The precision and accuracy of the Kerley and Ahlqvist-Damsten microscopic methods of age determination are compared. Both methods were applied to the same sample of 40 femoral thin sections of documented age at death. The results indicate that (1) both methods can be used with equal precision, as suggested by comparable observer errors; and (2) the Kerley method produces overall more accurate age estimates. The low previously published standard error of the Ahlqvist-Damsten method (6.71 years) apparently results from the uneven age distribution and small size (20) of their sample.     

Rapid stomatal responses to humidity

The response of leaf conductance in apple to rapid changes in atmospheric humidity was studied using a continuous flow porometer. Leaf-air vapour pressure difference was changed by adjusting the humidity of the inlet air or by altering the flow rate of the air through the chamber. The time course of the response of leaf conductance to leaf-air vapour pressure difference was monitored for periods up to 10 min using a chart-recorder. There were significant changes in leaf conductance within seconds of changing humidity. These were attributed to alterations in stomatal aperture.Abbreviations E evaporation rate - g leaf conductance - PAR photosynthetically active radiation     

Light and electron microscopic structure of golgi-stained neurons in the vertebrate brain (new rapid Golgi procedure)

Summary After application of a rapid, selective silver impregnation procedure for light (LM) and electron (EM) microscopy, individual neurons are distinguishable by a light silver precipitation. The silver content is sufficient that entire nerve cells can be observed light microscopically; on the other hand, electron microscopically the cytological details are still visible. Brains of mice were fixed by phosphate-buffered aldehyde perfusion, and pieces of tissue left in a 1 % K₂Cr₂O₇ solution for 13 h before impregnation in a 0.5 % AgNO₃ solution for 2h. Thick sections (30–50 m) of the impregnated tissue were cut; from these sections, suitably stained neurons were dissected out and re-embedded for ultrathin sectioning, thereby allowing observations on the same neurons at the EM level. A thin silver deposit was observed along the delimiting neuronal membrane, the microtubules and the smooth ER, including the spinal apparatus of the dendritic spines. The fine cytoplasmic details of the impregnated neurons and the surrounding tissue are well preserved and, therefore, suitable for subsequent determination of synaptic relationships of the impregnated neurons with the adjacent neuronal elements.     

The relationship between the nuage and the chromatoid body during spermatogenesis in the Rat

Summary Cytoplasmic structures ultrastructurally similar to the nuage are present in the cytoplasm of all spermatogenic cells in adult rats. The nuage is a discrete organelle which should not be confused with the chromatoid body. In step 7–8 spermatids transient contact is established between the nuage and the chromatoid body. This indicates a very specific recognition of the nuage by the chromatoid body. It is suggested that the nuage and the chromatoid body are separate cell organelles the functions of which are somehow related to each other.     

A toxic amino acid, 2(S)3(R)-2-amino-3-hydroxypent-4-ynoic acid from the fungus Sclerotium rolfsii

An amino acid, lethal to New Hampshire chickens (LD₅₀, 150 mg/kg) was isolated from dried sclerotia of the fungus Sclerotium rolfsii (Sacc.). Purification of the rather unstable compound was effected on a cation exchange column by means of displacement chromatography and the amino acid was crystallised from 80% methanol. A structure was assigned to the compound on the basis of available chemical and physical data, namely 2(S),3(R)-2- amino-3-hydroxypent-4-ynoic acid. Confirmation of this structure was gained by direct and indirect synthetic procedures.     

Teucrins H1-H4, novel clerodane-type diterpenes from Teucrium hyrcanicum

The structure and stereochemistry of teucrins H1, H2, H3 and H4, four novel diterpenes from Teucrium hyrcanicum have been established by detailed studies of the PMR and ¹³C NMR spectra, CD results and chemical transformations. Teucrin H4, a minor constituent is shown to possess a unique stereochemistry hitherto not reported for related diterpenoids.