Density Dependent Change of Myristoylated Proteins in C3H10T1/2 Fibroblasts and Their Transformants

We have examined the pattern of protein myristoylation in C3H10T1/2 fibroblasts during cell growth. During the growing phase of 10T1/2 cells, several proteins were radiolabelled with [³H]myristate, and among them proteins with molecular masses of 22, 35, a doublet of 42–45 and 67 kDa were labelled predominantly. The extent of myristoylation in each of these proteins changed with cell density. The amount of radioactivity incorporated into the 22 kDa protein in 10T1/2 cells decreased with increasing cell density and remained at a low level during the stationary phase. In contrast, the incorporation into the 67 kDa protein increased parallel to cell density. The density-dependent change of myristoylation was not observed in any of the transformants of 10T1/2 cells thus far examined. The 67 kDa protein was identified as MARCKS (myristoylated alanine-rich C kinase substrate) by immunoprecipitation with an anti-MARCKS antibody. By Western blot analysis, we found that the amount of MARCKS in 10T1/2 cells increased significantly analogous with cell density. Therefore, it is possible that MARCKS and the 22 kDa protein play a role in contact-mediated cell signalling in 10T1/2 cells, but the mechanism is lost in transformed cells. © 1997 John Wiley & Sons, Ltd.     

Effects of temperature on the development and survival of an endangered butterfly, Shijimiaeoides divinus barine (Leech) (Lepidoptera: Lycaenidae)

The effects of temperature on the development and survival of Shijimiaeoides divinus barine were examined in the laboratory in 2008. The eggs and larvae were reared at temperatures of 15, 17.5, 20, 25, 30 and 35°C with a long-day photoperiod of 16 h light : 8 h dark (16L : 8D). The highest hatchability of eggs was 88.0% at 20°C, but hatchability at high temperatures of 30 and 35°C was 30 and 0%, respectively. The lowest and highest survival rates from the first to third instar were 18.8% at 15°C and 76.9% at 20°C. Few deaths were observed after the fourth instar. The shortest developmental periods of the eggs and larvae were 4.0 and 15.8 days at 30°C, and the durations of the egg and larval stages increased significantly as the temperature decreased. The developmental zero and thermal constants were 9.6°C and 82.6 degree–days for the egg stage, and 10.7°C and 306.8 degree–days for the larval stage. The developmental period of the natural population of S. divinus barine in Azumino City, Nagano Prefecture was calculated using the developmental zero, thermal constants and Azumino City temperature data.     

Enzymic synthesis of floridean starch in a red alga, Serraticardia maxima

ADP-glucose: -l,4-glucan -4-glucosyltransferase was obtainedfrom a marine red alga Serraticardia maxima in a form boundwith floridean starch granules. The enzyme catalyzed the transferof glucosyl residue from ADP-glucose, UDP-glucose and GDP-glucoseto floridean starch added as a primer. ADP-glucose was the mostefficient glucosyl donor in the reaction. Maltose was producedby ß-amylolysis of the glucan synthesized by the algalenzyme. The optimum pH for enzyme activity was at 8.4. The enzymewas not obtained in a soluble form from either the chloroplastextract or the whole algal cell extract. Electron micrographsof algal cells revealed that floridean starch granules are localizedexclusively outside chloroplasts. Hence, it appears that mostof the synthetase is present outside chloroplasts. ¹ Contribution from the Shimoda Marine Biological Station, TokyoKyoiku University, No. 202. This work was supported by a Grant-in-Aidfor Cooperative Research from the Ministry of Education, Japan. ² Present address: Department of Biology, Faculty of Science,Science University of Tokyo, Kagurazaka, Tokyo 162, Japan. ³ Present address: Laboratory of Biology, Faculty of Science,Toho University, Narashino, Chiba 275, Japan. (Received May 25, 1970; )     

Chemical Factors Affecting Palmelloid-Forming Activity of Chloroplatinic Acid on Chlamydomonas eugametos

In order to elucidate the physiological mechanism of palmelloid formation of the unicellular green alga Chlamydomonas eugametos by chloroplatinic acid, the factors causing inhibition of the palmelloid formation were studied in various media. Chloroplatinic acid lost its palmelloid-forming effect in the presence of nitrite ion and amines having a second functional group. Nitrate, potassium, and sodium ions and monofunctional amines were without influence on the effect of chloroplatinic acid. These results indicate that the active form of platinum (IV) may undergo ligand substitution, the resulting complex being unable to cause palmelloid formation.     

Descriptions of two new species of butterflies (Lepidoptera,Lycaenidae) from the South Sinai

Two new species of lycaenid butterflies, Strymonidia jebelia and Pseudophilotes sinaicus, are described from the high mountains of South Sinai. The foodplants and the eggs of both species are also described with notes on the habitats and adult behaviour. Their relationships with closely allied species and the possible evolutionary histories are discussed.     

Expression and Transmission of Wild-Type Pigmentation in the Skin of Transgenic Orange-Colored Variants of Medaka (Oryzias latipes) Bearing the Gene for Mouse Tyrosinase

Transgenic fish carrying a reconstructed mouse tyrosinase gene, mg-Tyrs-J, were produced by microinjecting the gene into the oocyte nucleus of an orange-colored variant of medaka (Oryzias latipes). Of 64 oocytes microinjected and subsequently inseminated, 13 embryos developed normally beyond hatching and three of them exhibited brown skin pigmentation in the adult as was commonly observed in the wild type of this species. Light and electron microscopic examination disclosed a ubiquitous distribution of typical melanophores in the skin of these transgenic fish. Judging from their population density and distribution pattern, it was presumed that melanogenesis in these fish was elicited in amelanotic melanophores that resided in the skin of the orange-colored fish of this variant. Immunofluorescence with use of the anti-mouse tyrosinase antiserum lacking reactivity to medaka tyrosinase clearly disclosed that the gene introduced was expressed in the melanophores of transgenic fish. Crosses of female transgenic fish and males from an orange-colored variant yielded offspring exhibiting wild-type or orange-colored pigmentation in a ratio of 1:1, thus implying that mg-Tyrs-J integrated into the medaka genome behaves like a dominant gene. Little melanogenesis was observed in xanthophores, leucophores and iridophores in transgenic fish, suggesting possible specificity in recognition of teleostean cell types (i.e., melanophores) by the regulatory region of the mouse tyrosinase gene.     

Ten microsatellite loci from Zamia integrifolia (Zamiaceae)

Ten microsatellite loci isolated from Zamia integrifolia are described. All 10 are polymorphic, with three to 10 alleles across 36 members of a single population from South Florida. Heterozygosities ranged from 0.139 to 0.889. Two loci depart significantly from Hardy–Weinberg equilibrium, and exhibit heterozygote deficiency. One locus pair exhibits significant linkage disequilibrium. The primers have also successfully amplified loci from Zamia portoricensis and Zamia ambliphyllidia. These loci will be utilized for population studies in the Caribbean Zamia pumila complex.     

The accuracy of endometrial cytology in the diagnosis of endometrial adenocarcinoma

tajima m., inamura m., nakamura m., sudo y. and yamagishi k. (1998) Cytopathology 9 , 369–380
The accuracy of endometrial cytology in the diagnosis of endometrial adenocarcinoma
We have examined 62 234 cytological samples of endometrium to establish the accuracy and false-positive rate for the diagnosis of endometrial adenocarcinoma. The patients were either attending the gynaecological out-patients clinic with symptoms or were asymptomatic women attending for routine population screening as part of our cancer detection programme, the numbers from these two sources being equal. Out of 138 cases identified as endometrial adenocarcinoma by cytology 126 (91.3%) were confirmed histologically in our hospital. Twelve cases (8.7%) were shown to be false-positives. Re-examination of these led to the same false-positive diagnosis in all 12 cases. This was attributable to similarities of nucleo–cytoplasmic ratio, irregular arrangement of nuclei, variation in nuclear shape and in the numbers of nucleoli in repair cells and hyperplastic cells compared with the carcinoma cases. Most of the false-negative reports were due to insufficient material, pale staining in malignant cells or diagnostic error. Refraction measurement of the density of nuclei of cancer cells using equipment for which the patent is pending enabled objective measurement of nuclear density which indicated that the nuclei were not stained darkly enough in false-negative cases.