THE HUMAN CHROMOSOME : Electron Microscopic Observations on Chromatin Fiber Organization

Human lymphocytes were grown in short-term tissue culture and were arrested in metaphase with Colcemid. Their chromosomes were prepared by the Langmuir trough-critical point drying technique and were examined under the electron microscope. In addition, some chromosomes were digested with trypsin, Pronase, or DNase. The chromosomes consist entirely of tightly packed, 240 ± 50-A chromatin fibers. Trypsin and Pronase treatments induce relaxation of fiber packing and reveal certain underlying fiber arrangements. Furthermore, trypsin treatment demonstrates that the chromatin fiber has a 25–50 A trypsin-resistant core surrounded by a trypsin-sensitive sheath. DNase digestion suggests that this core contains DNA.     

Simple Medium for Pigment Production by the Erythrasma Diphtheroid

A simple method for the isolation and identification of the causative diphtheroid of erythrasma, Corynebacterium minutissimum, utilizes Muiller-Hinton Agar.     

Isolation of Shigellae: V. Comparison of Enrichment Broths with Stools

Many enteric media are more efficient for the detection of salmonellae than of shigellae. Comparisons of three enrichment broths and three plating media were made during analysis of 1,405 stool specimens to choose a combination of media which would enhance detection of shigellae as well. Gram-Negative (GN), Selenite, and Silliker's Broths were streaked to E M B, Salmonella-Shigella (SS), and xylose lysine deoxycholate (XLD) Agars. The enrichment broths produced a twofold increase in isolations of both salmonellae and shigellae over direct streaking. All three broths performed equally well for Salmonella detection, but GN and Silliker's produced twice as many Shigella isolates as did Selenite. Comparison of the plating media showed that XLD was markedly more efficient than either E M B or SS Agar for the recovery of both genera. SS Agar was superior to E M B for isolation of salmonellae after enrichment, whereas E M B was better for isolation of shigellae by direct streaking. Both E M B and SS were more effective when used after GN and Silliker's than after Selenite. GN Broth and XLD Agar were the most efficient combination of media. During these analyses, 158 salmonellae and 49 shigellae isolates were obtained.     

Certification of Probability of Sterilization of Liquid by Filtration

Four types of hydrosol filters, two reusable (diatomaceous cylinder and fritted-glass funnel) and two disposable (asbestos pad and membrane filter) were challenged with a heavy bacterial suspension to assess their ability to produce sterile filtrates. Two of the four diatomaceous earth filters, the four fritted-glass funnels, and all of the asbestos pads tested generally gave sterile filtrates. However, only one type of filter, one of the membranes in its manufacturer's own holder, consistently gave sterile filtrates. The two other types of membranes usually gave sterile filtrates if tested in one manufacturer's holder, but all types invariably gave contaminated filtrates when tested in another manufacturer's holder. Contaminated filtrates were generally attributed to a poor reusable filter or to a faulty holder used with a disposable filter. If a high degree of certainty is required for sterile heat-labile filtrate, it is suggested that the liquid be passed through two or more filters in a previously tested and proven system.     

Some Properties of Heat-Resistant and Heat-Sensitive Strains of Clostridium perfringens I. Heat Resistance and Toxigenicity

Heat resistance at 100 C (D-values), sporulating ratios, toxigenicity for mice, and lecithinase activity (as micrograms per milliliter of enzyme, ascertained by the lecithovitellin reaction) were determined for four strains of Clostridium perfringens. A definite inverse relationship between thermal resistance and toxigenicity was found. The D-values ranged from 17.6 for the most heat-resistant strain to 0.3 for the strain possessing the least heat resistance, with corresponding lecithinase activities from 25 to 133 mug/ml of enzyme. The sporulating ratios did not differ greatly between the strains. The heat stability of the toxin was greater at 100 C than at 75 C. There was a noticeable difference between the heat stabilities of the toxin in the culture fluids of the heat-sensitive and heat-resistant strains at pH 7.0 when the toxic filtrates were held at 100 C. At a holding temperature of 75 C, a similar but lesser difference was observed at pH 5.5. Heat resistance and lecithinase activity did not change when a substrain of the least heat-resistant parent strain was obtained through heat selection by a single transfer, or when the most heat-resistant strain was transferred serially 12 times.     

The incidence of double-yolked eggs in relation to improvement in egg production

Summary The production of double-yolked eggs and its relation to other egg production traits has been summarized over a period of thirty years for a closed flock selected for gains in total egg production.The number of double-yolked eggs per pullet as well as the percentage of pullets laying at least one double-yolked egg have increased rather steadily, although it is evident that the trait possesses no selective advantage. Pullets which laid double-yolked eggs showed earlier sexual maturity and superiority in egg production but it is clear that the corresponding genetic correlations are low or negligible.The heritability of the trait increased with its level of incidence and is sufficiently high that selection should increase the incidence to a level permitting further study of multiple ovulation.

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Zusammenfassung An einer für eine Steigerung der Ei-Produktion ausgewählten Herde Weißer Leghornhühner wurde der Anfall von Eiern mit zwei Dottern und ihre Beziehung zu anderen Merkmalen der Ei-Produktion für einen Zeitraum von 30 Jahren zusammengestellt.Die Anzahl doppeldottriger Eier je Hühnchen und der Prozentsatz Hühnchen, die wenigstens ein doppeldottriges Ei legten, ist ziemlich konstant geblieben, wenn auch dieses Merkmal offensichtlich keinen Selektionswert hat. Hühnchen, die Eier mit zwei Dottern legten, zeigten eine frühere Geschlechtsreife und eine höhere Ei-Produktion, aber es ist klar, daß die entsprechenden genetischen Korrelationen niedrig sind.Die Erblichkeit des Merkmals stieg mit seinem Auftreten, sie ist hoch genug, daß eine Selektion das Auftreten noch steigern könnte, um eine weitere Untersuchung der mehrfachen Ovulation zu ermöglichen.

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Dedicated to Professor Hans Stubbe on the occasion of his 65^th birthday.     

Golgi apparatus isolation and use in cell-free systems

Summary Recent advances in understanding the molecular mechanisms of membrane traffic to and through the Golgi apparatus have been predicated in large measure on the use of permeabilized animal cells, and on completely cell-free systems. These systems have included those addressing inter-Golgi apparatus membrane traffic, endoplasmic reticulum to Golgi apparatus traffic, and endocytotic events. Development of cell-free systems depends on the use of isolated fractions. Specificity is often achieved by using a compartment-specific assay so that the fractions employed can be very crude. More recently cell-free systems also have evolved which employ highly purified and well-characterized cell fractions. The latter may be utilized in the absence of a compartment-specific assay but may require employment of compartment-specific assays for validation. Central to development of cell-free systems for membrane analysis has been the availability of isolated Golgi apparatus, first from plants and later from animal tissues and cells. A major advantage of cell-free systems is that they are most clearly amenable to the investigation of molecular mechanisms of membrane trafficking.Dedicated to Hilton H. Mollenhauer on the occasion of his retirement