Catechol-O-methyltransferase: A method for autoradiographic visualization of isozymes in Cellogel

An electrophoretic procedure for separating the molecular forms of catechol-O-methyltransferase in cellulose acetate gel is described; the zones of enzyme activity were revealed by autoradiography. The electrophoretic patterns of the enzyme in several tissues and cell lines derived from four different species are presented.This investigation was supported by Grant 500.6/Ric. 70/1981 from the Italian Ministry of Health and by the Italian Ministry of Education. We are grateful to Dr. M. Castagnola for useful advice and help with thin-layer chromatography.     

Decreased triiodothyronine binding to isolated nuclei from livers of preobese and obese (ob/ob) mice

L-Triiodothyronine (T3) binding to hepatic nuclei from (ob/ob) mice at different ages was examined and compared with that of lean controls. Results showed a significant reduction in T3 binding in liver nuclei of obese mice at all ages studied. The preobese mice at 2 weeks of age had 27.9% fewer receptor sites/mg DNA compared to lean controls, receptor concentration further decreased to 67.7% at 18 weeks of age. Data presented here demonstrates that the impaired triiodothyronine (T3) binding to hepatic nuclei present in older (ob/ob) obese mice is an antecedent to the obesity. This report also helps to explain the poor thermoregulation and low oxygen consumption present during the preobese phase of the postnatal development of these animals.     

Electron paramagnetic resonance studies of heme c and its nitrosyl derivative in Vibrio (Achromobacter) fischeri nitrite reductase

Interactions of Vibrio (formerly Achromobacter) fischeri nitrite reductase were studied by electron paramagnetic resonance spectroscopy. The spectrum of the oxidized enzyme showed a number of features which were attributed to two low-spin ferric hemes. These comprised an unusual derivative peak at g = 3.7 and a spectrum at g = 2.88, 2.26, and 1.51. Neither heme was reactive in the oxidized state with the substrate nitrite and with cyanide and azide. When frozen under turnover conditions (i.e., reduction in the presence of excess nitrite), the enzyme showed the spectrum of a nitrosyl heme derivative. The g = 2.88, 2.26, and 1.51 signals reappeared partially on reoxidation by nitrite, indicating that the nitrosyl species which remained arose from the g = 3.7 heme. The nitrosyl derivative showed a 14N nuclear hyperfine splitting, Az = 1.65 mT. The nitrosyl derivative was produced by treatment of the oxidized nitrite reductase with nitric oxide or hydroxylamine. Exchange of nitric oxide between the nitrosyl derivative and NO gas in solution was observed by using the [15N]nitrosyl compound. A possible reaction cycle for the enzyme is discussed, which involves reduction of the enzyme followed by binding of nitrite to one heme and formation of the nitrosyl intermediate.     

Clinical and experimental keratitis caused by the Colletotrichum state of Glomerella cingulata and Acrophialophora fusispora

Two cases of mycotic keratitis caused by the Colletotrichum state of Glomerella cingulata and Acrophialophora fusispora are reported for the first time. Both the isolates produced experimental corneal lesions in rabbit eyes but A. fusispora was more pathogenic. The experimental infection was more severe, with both the fungi, in rabbits pretreated with cortisone as compared with untreated animals. In vitro A. fusispora was most sensitive to miconazole and tolciclate followed by clotrimazole, amphotericin B and lactones while clotrimazole exerted maximum inhibitory effect on Colletotrichum followed by miconazole, lactones, amphotericin B and arnebins. Arnebins and tolciclate were inactive respectively against A. fusispora and Colletotrichum. Of the 3 drugs tested in vivo, against A. fusispora keratitis in rabbit, amphotericin B showed better results than tolciclate and miconazole.     

Physiology of Morphactins: Effect on Gravi- and Photo-response

Plant roots and shoots respond to gravity and light source in a definite way. Thus, there are typical geotropic and phototropic responses for roots and shoots. When seedlings were grown in presence of morphactins, IT 3233 or IT 3456, on a vertical or a horizontal plane, the roots and shoots lost the capacity to respond to gravity or to unilateral light source. This was true for both monocots and dicots. This suggests that basic mechanism (s) of the two tropic responses are the same in the roots and shoots of the two plant groups. The site(s) of action of morphactins is unknown. The reaction (s) controlling geotropism and phototropism may be closely related as morphactins affected both geotropic and phototropic response of the same organ. Indoleacetic acid and gibberellic acid per se did not modify the effect of morphaclins on geotropism. Growth retardation effect of morphactins appears to be controlled by another mechanism.     

Structure of the bovine eye lens gamma s-crystallin gene (formerly beta s)

The organization of a number of crystallin genes has already been resolved. One of the remaining genes of which the structure was hitherto unknown is the gamma s gene (formerly beta s). We determined the complete sequence of the bovine gamma s-crystallin-coding gene, apart from the middle region of the first intron. Since it contains three exons and two introns, we conclude that the former beta s, also at the gene level is gamma-crystallin-like. However, it is located on chromosome 3, in contrast to other gamma genes which occur in tandem on the human chromosome 2.