The axolotl as an animal model for the comparison of 3-D ultrasound with plain film radiography.

We assessed the usefulness of an animal model, the axolotl (Ambystoma mexicanum), in comparing 3-D ultrasound (3-D US) and plain film radiographs. Hindlimbs were amputated from 5 animals, at either the zeugopodial or stylopodial level, and each regenerating limb was imaged 16 times with 3-D US and 14 times with plain film X ray over 315 days. US images were acquired with a Siemens Sonoline Versa Pro and a 10-MHz linear array transducer. For 3-D US images, the probe was translated in a motor-driven linear stage while images were digitized. The regenerating tibia and fibula bones were detected on 3-D US an average of 37 days earlier than on plain film radiography, and regenerating phalangeal bones were detected on 3-D US an average of 18 days earlier. After 120 days, both imaging modalities consistently showed the bones. The average bone growth rates for the tibia and fibula were 0.019 +/- 0.001 mm/day and 0.019 +/- 0.001 mm/day, respectively.

Surface contraction and expansion waves correlated with differentiation in axolotl embryos. II. In contrast to urodeles, the anuran Xenopus laevis does not show furrowing surface contraction waves.

We have observed a number of contraction waves traversing the axolotl (Ambystoma mexicanum) embryo (a urodelan amphibian) from the midblastula transition up to at least neural tube closure, and wished to learn if similar "differentiation waves" appear on the popular laboratory anuran amphibian, the South African clawed toad, Xenopus laevis. Time lapse video microscopy showed that no contraction waves are visible on the surface of Xenopus from gastrulation through neurulation. It is possible that cell intercalations in the double-layered ectoderm of the Xenopus embryo are homologous to the surface waves in the single layered ectoderm of the axolotl embryo. In any case, a simple, universal correspondence between surface waves and induction phenomena and differentiation does not exist.

Surface contraction and expansion waves correlated with differentiation in axolotl embryos--I. Prolegomenon and differentiation during invagination through the blastopore, as shown by the fate map.

We have discovered a series of expansion and contraction, solitary waves that correlate with discrete steps of differentiation in the urodele amphibian axolotl embryo (Ambystoma mexicanum). Here we examine in detail the proposition that the blastopore is a set of differentiation waves. We superimposed the image of the axolotl fate map onto our digitized video images of normal gastrulation and matched the fate map to pigmentation irregularities on the embryo. We were then able to track the invagination of the fate map by tracking the variegated pigmentation on several embryos as gastrulation proceeded. We show a particular expansion and contraction wave sequence for every tissue in the blastula stage fate map and can now explain precisely why the fate map has the shape it does and its relationship to the embryo at subsequent stages. Each tissue can be assigned a differentiation code and placed on a hierarchical, binary differentiation tree.

Furrowing surface contraction wave coincident with primary neural induction in amphibian embryos.

We predicted, and have now observed, a surface contraction wave in axolotl (Ambystoma mexicanum) embryos that appears to coincide temporally and spatially with primary neural induction and homoiogenetic induction, and with involution of the chordomesoderm. The wave starts from a focus anterior to the dorsal lip of the blastopore and spreads as an ellipse, until part of it encounters the rim of the blastopore and vanishes there. The remaining arc then continues over the dorsal hemisphere until it reforms an ellipse that decreases in size. About 9 to 12 hours after it begins, the wave vanishes at a focus diametrically opposite its point of origin. The wave involves both local contraction and furrowing in the monolayer ectoderm. To a good approximation, the hemispherical portion of the ectoderm traversed by the wave becomes neuroepithelium, while the ectoderm not traversed by the wave becomes epidermis. The wave might provide a mechanism to determine the time and location at which neuroepithelial differentiation occurs.

[Nuclear state splitting: a working model for the mechanochemical coupling of differentiation "waves" with the controlling genes (master genes)]. / Metastabil'noe sostoianie iadra: rabochaia model' mekhanokhimicheskoi sviazi "voln" differentsirovki s upravliaiushchimi genami (master genes).

The cell state splitter (CSS) model for differentiation, supported by recent evidence, requires that an intracellular mechanical events result in a commitment signal to the nucleus that determines which of two readied gene cascades is to be activated, via their master genes. We show how a link between the mechanochemistry of cell state splitting and the molecular genetics of "nuclear state splitting" might be forged out of well known molecular components of cells.

On the pulmonary tumorigenicity of arsenic trisulfide and calcium arsenate in hamsters.

The pulmonary tumorigenicity of arsenic trisulfide and calcium arsenate was explored following 15 intratracheal instillations in male Syrian golden hamsters at weekly doses of about 3 mg/kg body weight (as arsenic). One lung adenoma appeared in 28 animals examined in the arsenic trisulfide group and 4 adenomas in 35 animals surviving the treatment with calcium arsenate. No adenomas were seen in 26 animals instilled with the vehicle (0.9% saline solution). The results show that calcium arsenate is tumorigenic (P less than 0.05), while the evidence is inconclusive for arsenic trisulfide.

Carcinomas of the respiratory tract in hamsters given arsenic trioxide and/or benzo[a]pyrene by the pulmonary route.

The pulmonary carcinogenicity of arsenic trioxide (As) alone and in combination with benzo[a]pyrene (BP) was studied in male Syrian golden hamsters given 15 weekly intratracheal instillations. The animals were divided into four groups: As, BP, As + BP, and controls. At each instillation about 3 mg/kg body wt as arsenic and/or 6 mg/kg body wt of BP was administered. All groups received a carrier dust (charcoal carbon), which increased the lung retention of arsenic, as well as sulfuric acid. Carcinomas of the larynx, trachea, bronchi, or lungs were found in 3, 17, and 25 of 47, 40, and 54 animals examined in the As, BP, and As + BP groups. No respiratory tract carcinomas were found in 53 control animals. The incidence of pulmonary adenomas, papillomas, and adenomatoid lesions was markedly higher in the As group than in the control group (P less than 0.01). Taken together the data provide strong evidence that arsenic trioxide can induce lung carcinomas. Furthermore, there was some evidence of a positive interaction between arsenic and BP in relation to adenomatous lung tumors, which could be of importance for the synergism between arsenic and smoking observed in smelter workers.

Ochratoxin A as the cause of spontaneous nephropathy in fattening pigs.

At a number of slaughters nephropathy and high ochratoxin A contents in kidneys have been observed in fattening pigs from two Swedish farms. In one herd the source of contamination was barley grown on the home farm and stored under such conditions that the growth of fungal species (Penicillium verrucosum var. verrucosum) producing ochratoxin A occurred, with the subsequent formation of the toxin. In this case high ochratoxin A levels in fattening pigs were found during a period of about 18 months. In the second herd, where compounded feed was used, it was impossible to locate the source of contamination. It was presumed that a consignment of feed was damaged by rain during storage at the farm. Ochratoxin A was found in fattening pigs from this herd for a period of about 2 months. Ochratoxin A appeared in the kidneys of all investigated pigs. In some animals the livers, whole blood, and plasma were analyzed, too. The livers contained somewhat lower amounts of ochratoxin A than the kidneys, whereas the content in whole blood and plasma, respectively, was 5 and 13 times greater. Kidneys spontaneously contaminated with ochratoxin A, when stored for 10 months at -70 degrees C, showed no systematic decrease in toxin content.