Electrophoretic separation of multiple bands with beta-glucuronidase activity in human sera

An electrophoretic procedure is described which resulted in the separation of up to four bands with beta-glucuronidase activity in human sera. Initial screening of a small sample of individuals revealed persons with two, three, and four bands.Supported by Special Project Grant Number 414 of the Children's Bureau, U.S. Department of Health, Education and Welfare.     

Melanogenesis in amphibians

Summary The pigmented epithelium of Rana pipiens tadpole eyes normally develops at least two types of melanosomes: (1) an elongated melanin granule of relatively homogeneous electron density, and (2) a complex melanosome which has an outer electrondense area and one or more less dense cores. Evidence indicates that complex melanosomes are formed by new melanin enclosing preexisting melanosomes. An organized fibrillar premelanosome is demonstrated with the aid of the antimelanogenic compound phenylthiourea (PTU). These premelanosomes are the developing forms of the elongated melanosomes. There is evidence that the premelanosomes originate in the smooth endoplasmic reticulum. Phenylthiourea blocks melanin synthesis in the premelanosomes; however, removal of the PTU allows pigment deposition. This finding of an organized, fibrillar premelanosome in an amphibian marks the lowest phylogenetic group in which these organelles have been described.An Oak Ridge Graduate Fellow from Catholic University of America, Washington, D.C., under appointment from Oak Ridge Associated Universities.The MAN Program is supported by the National Cancer Institute, the National Institute of General Medical Sciences, the National Institute of Allergy and Infectious Diseases, and the U.S. Atomic Energy Commission.Oak Ridge National Laboratory is operated by Union Carbide Corporation Nuclear Division for the U.S. Atomic Energy Commission.     

Assimilation of Alkanes and Alkenes by Fungi

A group of filamentous fungi were assayed for their ability to utilize a series of n-alkanes and 1-alkenes as the sole source of carbon. Although strains of Cunninghamella exhibited profuse growth on most of the hydrocarbons tested, the majority of fungi tested were able to produce definite growth on one or more of the compounds. The hydrocarbons with a 14-carbon chain length appeared to be more consistently utilized than any other. Strains of Aspergillus appeared to differ in their capacity to utilize individual members of the hydrocarbon series. Thin-layer chromatographic analyses of ether extracts from C. blakesleeana grown on n-tetradecane and 1-tetradecene were similar and revealed the presence of a monocarboxylic acid, a primary alcohol, and a secondary alcohol.     

Thirst and artificial heat acclimatization in man

The purpose of this study was to investigate the relationship between serum osmotic changes, water intake and water balance in four, fit young men during and after exercise in the heat, before and after artificial heat acclimatization. During exercise, before steady-state conditions were reached, voluntary water intakes generally paralleled but were not proportional to the serum osmotic pressure. In steady-state conditions, drinking was approximately proportional to the effective osmotic pressure of the serum. During the post-exercise recovery period, when serum osmolarity returned to normal levels, water intake also subsided even though there was a total body water (wt) deficit of about 3%. Body weight did not return to control levels until 62 to 86 hr following the exercise. This slow recovery could not be accounted for by a loss of water associated with glycogen utilization during exercise or by sweat electrolyte depletion. In general, the results supported Dill's hypothesis but plasma volume changes, in addition to osmotic factors,are very likely operative in the initiation and satiation of drinking under these conditions. Perhaps slower acting volume control mechanisms mediate the slow recovery of total body water.

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Zusammenfassung Untersucht wurde die Beziehung zwischen osmotischen Veränderungen im Serum, Wasseraufnahme und Wassergleichgewicht bei 4 gesunden,jungen Männern während und nach körperlicher Arbeit in der Hitze vor und nach künstlicher Hitzeakklimatisation. Die freiwillige Wasseraufnahme während der Arbeit, ehe ein steady state erreicht wurde, war im allgemeinen gleichlaufend, aber nicht proportional dem osmotischen Serumdruck. Im steady state war die Trinkmenge ungefähr proportional dem effektiven osmotischen Serumdruck. Wenn die Serumosmolarität während der Erholungsphase zur normalen Werten zurückkehrte, liess die Wasseraufnahme ebenfalls nach,obwohl das Gesamtkörperwasserdefizit ungefähr 3% betrug.Die Körpergewichte erreichten die Ausgangswerte 62–86 Std nach der Arbeit. Diese langsame Erholung war nicht bedingt durch einen Wasserverlust dur Glykogenverwertung während der Arbeit oder durch Elektrolytverluste beim Schwitzen. Die Ergebnisse stützen Dill's Hypothese, doch unter diesen Bedingungen wirken Plasmavolumen-Veränderungen zusätzlich zu osmotischen Faktoren sehr wahrscheinlich mit bei der Auslösung des Trinkens und der Sättiging mit Wasser. Vielleicht vermitteln langsamer wirkende Volumenkontrollmechanismen die langsame Wiederherstellung der Gesamtkörper-Wasservorräte.

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Resume On a examiné les relations existant chez 4 jeunes hommes sains entre les variations de la pression osmotiques du sérum d'une part,l'ingestion d'eau et l'équilibre acqueux du corps d'autre part et cela pendant et après un travail corporel à la chaleur tant avant qu'après une acclimatisation artificielle au chaud.L'ingestion volontaire d'eau évolue similairement à la pression osmotique du sérum, mais ne lui est pas proportionnel. Ce phénomène s'observe jusqu'à ce qu'un état dit stationnaire soit atteint. Dès ce moment, les quantités bues sont à peu près proportionnelles à la pression osmotique effective. Pendant la phase de récupération, la pression osmotique redevient normale et l'eau ingérée diminue bien que le déficit en eau pour le corps entier soit encore de 3% environ.Le poids du corps ne retrouve sa valeur initiale que 62 à 86 heures après la période de travail. Ce lent rétablissement n'est pas dû à des pertes d'eau par suite de l'utilisation de glycogènes pendant l'exercice ou par suite de la perte d'électrolytes par la sueur. Ces résultats viennent confirmer les hypothèses de Dill. Pourtant, dans ces conditions, les variations de volume du plasma agissent très vraisemblablement en plus des facteurs osmotiques dans le besoin de boire ou l'état de satiété. Il est possible que des mécanismes de contrôle du volume agissant plus lentement président aux phénomènes de rétablissement des réserves en eau dans son ensemble.

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Results were presented in part at the "Symposium on Nutrition and Physical Activity", The Swedish Nutrition Foundation, Tylösand, Sweden, 15–17 August,1966.     

Protein Components of Bacteriophages λ and λ Virulent

Parallel studies have been made of the protein coats of the temperate bacteriophage λ and of a deletion mutant, λ virulent. A new method for preparing ghosts of both phages by the action of Cu⁺⁺ is described. Protein ghosts of both phages can be dissolved in citrate at pH values below 3, more rapidly in the presence of 8 m urea. Both phages yielded three apparently identical protein components which can be separated by thin-layer gel filtration and thin-layer gel electrophoresis. The protein of molecular weight 47,000 ± 1,500 represents about 55% of the protein of the ghosts and is therefore likely to be the subunit of the head. The other proteins of molecular weight 30,000 ± 1,500 and 16,000 ± 1,500 represent approximately 25% and 20% of the protein, respectively. Amino acid analyses of the ghosts from the two phages have been carried out and show no significant differences. The buoyant density of phage λ virulent is 0.016 g/ml less than that of λ. Since no differences have been found in the protein components of the two phages, this indicates that the virulent mutant contains approximately 16% less deoxyribonucleic acid than the temperate phage.     

Ventricular septal defect with bidirectional shunting: Mathematical considerations

Previously proposed formulae for the quantitative estimation of bidirectional shunts across ventricular septal defects require determination of the oxygen contents of mixed venous, pulmonary artery, pulmonary venous, and aortic blood. Because these formulae do not take into account the mixing of oxygenated with unoxygenated blood within the ventricles, their use must result in underestimation of shunt flows in each direction. A mathematical model for a ventricular defect is examined, in which it is assumed that mixing of blood occurs in each of six sites in the venae cavae or right atrium, right ventricle, pulmonary artery, left atrium, left ventricle, and aorta. A total of fourteen streams of blood can flow from one to another of these mixing sites. As long as complete mixing occurs in the six specified mixing sites, any degree of mixing or non-mixing of the various streams is permitted. From the equations characterizing the model, formulae are derived in which the shunt flow in each direction is expressed in terms of the oxygen contents in the six mixing sites and the fractions of blood which enter the shunt from either side without prior mixing in a ventricular mixing site. The previously reported formulae, which apply when no ventricular mixing is allowed to occur, lead to theoretical minimum values for the shunt flows in each direction. At the opposite extreme where all the shunting blood is required to mix in a ventricle before entering the shunt, formulae for maximum possible shunt flows are also obtained. The absolute values for the left-to-right and right-to-left shunt flows, which must lie somewhere between the theoretical maximum and minimum values, cannot be computed from blood gas data alone. This work was supported in part by grant HE-07563 from the National Heart Institute of the National Institutes of Health and grants-in-aid from the American and North Carolina Heart Associations and the Life Insurance Medical Research Fund. Work completed during tenure as U.S.P.H.S. post-doctoral fellow.     

Survival and Recovery of Enterovirus from Foods

Type 1 poliovirus and type B1 and B6 coxsackieviruses added to eight commercial frozen or convenience foods before storage at room temperature, 10 C, and -20 C were still viable after various intervals of time up to 1 week, 1 month, and 5 months, respectively. Infectivity titers were determined in monkey kidney tissue culture. Decomposition which took place in food stored at room temperature did not affect the survival of virus, and antibiotics controlled bacterial growth during assay. A rapid, significant reduction in type B6 coxsackievirus was observed in cole slaw. Preliminary data indicate that sodium bisulfite could be the responsible ingredient. A less rapid reduction of type 1 poliovirus and type B6 coxsackievirus was found in hashed brown potatoes when stored at 10 C.