bluuebunny/crossref_metadata_2025 huggingface dataset:
Up‐regulation of a photosystem II core protein phosphatase inhibitor and sustained D1 phosphorylation in zeaxanthin‐retaining, photoinhibited needles of overwintering Douglas fir
Authors: EBBERT V. , ADAMS III W. W. , MATTOO A. K. , SOKOLENKO A. , DEMMIG‐ADAMS B.
Published: January 2005

Overwintering needles of the evergreen conifer Douglas fir exhibited an association between arrest of the xanthophyll cycle in the dissipating state (as zeaxanthin +  antheraxanthin; Z + A) with a strongly elevated predawn phosphorylation state of the D1 protein of the photosystem II (PSII) core. Furthermore, the high predawn phosphorylation state of PSII core proteins was associated with strongly increased levels of TLP40, the cyclophilin‐like inhibitor of PSII core protein phosphatase, in winter versus summer. In turn, decreases in predawn PSII efficiency, Fv/Fm, in winter were positively correlated with pronounced decreases in the non‐phosphorylated form of D1. In contrast to PSII core proteins, the light‐harvesting complex of photosystem II (LHCII) did not exhibit any nocturnally sustained phosphorylation. The total level of the D1 protein was found to be the same in summer and winter in Douglas fir when proteins were extracted in a single step from whole needles. In contrast, total D1 protein levels were lower in thylakoid preparations of overwintering needles versus needles collected in summer, indicating that D1 was lost during thylakoid preparation from overwintering Douglas fir needles. In contrast to total D1, the ratio of phosphorylated to non‐phosphorylated D1 as well as the levels of the PsbS protein were similar in thylakoid versus whole needle preparations. The level of the PsbS protein, that is required for pH‐dependent thermal dissipation, exhibited an increase in winter, whereas LHCII levels remained unchanged.
Genetic diversity for drought resistance in wild emmer wheat and its ecogeographical associations
Authors: PELEG Z. , FAHIMA T. , ABBO S. , KRUGMAN T. , NEVO E. , YAKIR D. , SARANGA Y.
Published: January 2005

Wild emmer wheat (Triticum turgidum spp. dicoccoides (Körn.) Thell.), the tetraploid progenitor of cultivated wheat, is a potential source for various agronomical traits, including drought resistance. The objectives of this study were to characterize (1) the genetic diversity for drought resistance in wild emmer wheat, and (2) the relationship between drought responses of the wild emmer germplasm and the ecogeographical parameters of its collection sites. A total of 110 wild emmer accessions consisting of 25 populations and three control durum wheat cultivars were examined under two irrigation regimes, well‐watered (’wet’) and water‐limited (’dry’). Wide genetic diversity was found both between and within the wild emmer populations in most variables under each treatment. A considerable number of the wild emmer accessions exhibited an advantage in productivity (spike and total dry matter) over their cultivated counterparts. Most wild emmer wheat accessions exhibited a greater carbon isotope ratio (δ13C, indicating higher water‐use efficiency) under the dry treatment and higher plasticity of δ13C relative to the cultivated controls, which may have contributed to the drought adaptations in the former. The most outstanding drought‐tolerance capacity (in term of productivity under the dry treatment and susceptibility indices) was detected in wild emmer populations originated from hot dry locations. The results suggest that wild emmer has the potential to improve drought resistance in cultivated wheat.
Characterization of the wound‐inducible protein ipomoelin from sweet potato
Authors: CHEN Y.‐C. , CHANG H.‐S. , LAI H.‐M. , JENG S.‐T.
Published: January 2005

The ipomoelin (IPO) gene, a wound‐ and methyl jasmonate‐inducible gene, was isolated from sweet potato (Ipomoea batatas cv. Tainung 57), and previously demonstrated to be regulated by dephosphorylated proteins and calcium ion (Chen Y.‐C. et al. Plant Cell and Environment 26, 1373–1383, 2003). In this report, the function of the IPO protein was further studied. The IPO gene was characterized as having one intron and presenting two copies within the genome of sweet potato. The IPO protein appeared 1 d after the leaves of sweet potato were wounded. Surprisingly, the accumulation of the IPO protein remained for 7 d after wounding. Additionally, after the IPO protein was fused to a histidine tag, the His‐IPO fusion protein produced from Escherichia coli BL21DE3 was then used to perform the haemagglutination test, which demonstrated that His‐IPO fusion protein agglutinated human blood cells. Furthermore, several carbohydrates, including methyl α‐ d‐glucopyranoside, methyl α‐ d‐mannopyranoside, maltose, mannose, glucose, galactose, and lactose, reduced the efficiency of the His‐IPO fusion protein in agglutinating human blood cells. These experimental results may indicate that the IPO protein is a lectin, a carbohydrate‐binding protein. Notably, the IPO protein retarded the growth and development of silkworm, and thus reduced silkworm survival rates. Therefore, these findings indicate that the function of the IPO protein is to protect plants from insect attack.
Golfer's vasculitis
Authors: Kelly Robert I , Opie Jacinta , Nixon Rosemary
Published: January 2005

A number of patients presented with an erythematous, purpuric rash occurring on the legs in association with playing golf and also after prolonged walks or hikes. Many patients believed that it was an allergic reaction to grasses or insecticides and had sometimes undergone extensive allergy testing. We collected reports of 17 such cases from dermatologists in the state of Victoria, Australia. Patients were interviewed by phone and asked to submit photographs of the rash if possible. Of these, the eruption developed in 15 after playing 18 holes of golf and in three following prolonged hikes. The rash would usually develop over the summer months under hot conditions. Most patients were over 50 years of age when the tendency to develop the eruption began. Biopsies of the rash in the active phase showed leukocytoclastic vasculitis. Patch testing and investigations for potential underlying causes for vasculitis were negative or unremarkable. It would seem that this is a common but poorly documented condition. The clinical presentation and histology would support the conclusion that it represents a leukocytoclastic vasculitis induced by prolonged exercise under hot conditions. The findings would suggest that it occurs in healthy people and extensive investigation with blood tests or allergy testing is inappropriate. We believe the condition should be termed ‘golfer's vasculitis', as golf appears to be the most common precipitating event and such a term would enable the condition to become more widely recognized.
Complicated lichenoid drug eruption
Authors: Armour Katherine , Lowe Patricia
Published: January 2005

We report a case of severe lichenoid drug eruption with multiple possible causative agents. A hepatitis C‐positive male presented with a short history of painful erosions of the vermilion, lichenoid lesions on the buccal mucosa and glans penis, and erosions and lichenification of the scrotum. In addition, he had a pruritic polymorphic eruption over the scalp, trunk and limbs, comprising psoriasiform and eczematous lesions. He had received combination therapy of pegylated interferon‐α‐2a and ribavirin, along with granulocyte colony‐stimulating factor for interferon‐induced leucopenia, and propranolol for portal hypertension. The former three agents were ceased 3 weeks prior to presentation, but he remained on propranolol at the initial dermatology consultation. The polymorphous clinical picture was consistent with lichenoid drug eruption, which was confirmed on histology. The papulosquamous eruption responded quickly to 2 weeks of oral prednisone 25 mg daily, which was tapered to 1 mg over 3 months and then ceased. The mucosal lesions were slow to improve and required the addition of tacrolimus 0.03% solution t.d.s. for complete resolution.
Copper‐induced cutaneous sarcoidosis
Authors: Masel Grant
Published: January 2005

A 52‐year‐old woman presented with gradual painless enlargement of both earlobes. Indurated plaques were also present elsewhere. There was no evidence of a systemic granulomatous disorder. Histopathology showed foreign material associated with granulomatous inflammation. Scanning electron microscopy and energy‐dispersive analysis of X‐rays demonstrated foreign material composed primarily of copper. Deposition is likely to have been related to corrosion of copper‐containing earrings, or from deposition at the time of ear piercing. The presence of clinically similar granulomatous lesions remote from the earlobes and not containing copper suggest the presence of underlying cutaneous sarcoidosis. The earlobe lesions are more likely to be caused by foreign matter acting non‐specifically as a nidus for cutaneous sarcoidosis, rather than by a specific pro‐inflammatory effect of dermal copper. After 3 months, the lesions have partially responded to oral hydroxychloroquine 200 mg/day.
Atypical presentation of erythema elevatum diutinum
Authors: Ly Hoang , Black Martin M
Published: January 2005

A 47‐year‐old man presented with two red‐brown plaques on his back. The histology was consistent with erythema elevatum diutinum. The case was unusual in the site of presentation and in the paucity of lesions. Investigations were aimed at screening for associations such as infections, malignancies and inflammatory diseases. Dapsone remained an effective treatment.
EMBRYO AND ORGAN CULTURES OF THE HORSESHOE CRAB, TACHYPLEUS TRIDENTATUS*
Authors: YAMAMICHI YOSHIO , SEKIGUCHI KOICHI
Published: January 2005

This paper deals with some preliminary technical experiments on cultures of the embryos and organs of Tachypleus tridentatus. In embryo culture, using the embryos at various stages of development, embryonic areas with hardly any yolk remaining in them were cultured in various media and by various methods (Series A); hemispheres of embryos containing the embryonic area (Series B) and whole embryos without egg membranes (Series C) were cultured in sterilized sea water. The suitability of the culture media and methods is described; the medium containing calf serum or 199, and the hanging‐drop method were found to be the most appropriate of those used in respective series of experiments. The changes in external and internal structures of the explants from embryos at various developmental stages cultured in various media and by various methods are described; dorsal closure, secretion of inner egg membrane, thickening of the cell layer and the elongation of limb buds were all observed in the Series A experiments. In Series B and C, coelom formation and also normal development of the embryos cultured after the 2nd embryonic moulting were observed. In organ culture, fragments of the embryonic heart were cultured in various media and by various methods. The hanging‐drop method resulted in longer survival than the method using agar media.
CHANGE IN THE ACTIVITY OF ARYLESTERASE IN SEA URCHIN EGGS FOLLOWING FERTILIZATION
Authors: MITA M. , YASUMASU I.
Published: January 2005

The activity of arylcsterase in sea urchin eggs (Anthocidaris craxsispina), increases at 5 min after fertilization to about 1.5‐fold that in unfertilized eggs, and decreases at 15 min to a lower level than that in unfertilized eggs. Then the activity of the enzyme increases again. The enzyme activity in unfertilized eggs is enhanced by either fructose 1, 6‐diphosphate (FDP) at concentrations between 4 and 10 μM, or guanosine 3′, 5′‐cyclic monophosphalc (cGMP) at concentrations between 0.1 and 0.3 μM. The activity is detectable in the crude microsomal fraction and also in the supernatant fraction obtained from sea urchin egg homogenates by centrifugation at 105,000 × g for 2 hr.
DIFFERENTIATION OF THE DIGESTIVE TRACT EPITHELIUM UNDER THE INFLUENCE OF THE HETEROLOGOUS MESENCHYME OF THE DIGESTIVE TRACT IN THE BIRD EMBRYOS
Authors: YASUGI SADAO , MIZUNO TAKEO
Published: January 2005

The endoderm of the oesophagus, proventriculus, gizzard or small intestine of the 5‐day‐old chick or quail embryo was cultivated in combination with homologous or heterologous mesenchyme on a WxxxOLFFyyy and HxxxAFFHNyyy medium for 7 to 21 days or on the chorio‐allantoic membrane (CAM) for 8 days. With homologous mesenchyme the epithelium always differentiated homotypically. In association with heterologous mesenchyme, the differentiation of the epithelium was both homotypical and heterotypical depending on the region of the digestive tract. The oesophagus and small intestine differentiate mainly homotypically both in culture and on CAM, but the gizzard and proventriculus show heterotypic differentiation particularly on CAM. Thus, the endoderm of the digestive tract of the 5‐day‐old chick or quail embryo, though rather “determined”, still reacts to the heterologous stimuli of the mesenchyme to some degree.
INDUCTION OF MYOCARDIOGENESIS FROM MOUSE TERATOCARCINOMA
Authors: UNO KAZUKO , AMANO SHIGETOYO
Published: January 2005

Cells of the isolated embryoid body line, SEBIlI, differentiate predominantly into pulsating cardiac muscle cells in vitro, after intraperitoneal passage before the cells are transferred into the culture medium. Quantitative analysis of myocardiogenesis in vitro revealed that the intraperitoneal passage induces myocardiogenesis in the embryoid bodies, while culturing in vitro after the passage is related to the occurrence of spontaneous pulsation which is the phenotypic expression of cardiac muscles. The possibility that this teratocarcinoma system offers a way of studying the control mechanism operating in normal embryonic myocardiogenesis is discussed.
PERIODIC CHANGE IN THE TENSION AT THE SURFACE OF ACTIVATED NON‐NUCLEATE FRAGMENTS OF SEA‐URCHIN EGGS
Authors: YONEDA MITSUKI , IKEDA MARIKO , WASHITANI SETSUKO
Published: January 2005

Unfertilized eggs of the sea urchin, Hemicentrotus pulcherrimus, were separated into two fragments by centrifugal force. The enucleate fragment (merogone) was subsequently activated by treating it with butyric acid and the tension at the surface was continuously measured by a compression method. The activated merogone was found to exhibit cyclic changes in tension, with a temporal pattern very similar to that of the changes accompanying the division cycle of normally fertilized eggs. This indicates quantitatively the presence in the cytoplasm of some periodic activity which can be triggered without nuclear control. Further, a periodic thickening of the intrahyaloplasmic space of the activated merogone, as noted by Kojima (14), was confirmed on the basis of extended observation.
ORGANIZATION AND REPLICATION ACTIVITY OF THE MITOCHONDRIAL MASS OF OOGONIA AND PREVITELLOGENIC OOCYTES IN XENOPUS LAEVIS
Authors: TOURTE MONIQUE , MIGNOTTE FRANÇOISE , MOUNOLOU JEAN‐CLAUDE
Published: January 2005

The origin of the mitochondrial mass, previously well characterized in Xenupus diplotene oocytes, has been traced up to oogonia by means of electron microscopy. A polarized organization of the oogonia and of the oocytes of the succeeding stages was observed. The mitochondrial cloud was found to be built up in the centriolar region near the site where the chromosomes will be implanted along the nuclear envelope at the “bouquet” stage. Autoradiographic studies of thymidine incorporation into mitochondrial DNA suggest that mitochondrial DNA synthesis is active throughout this early period of oogenesis.
Electrical Properties of Toad Oocytes During Maturation and Activation
Authors: IWAO YASUHIRO , ITO SHIZUO , KATAGIRI CHIAKI
Published: January 2005

The full‐grown oocytes of the toad Bufo bufo japonicus, whether in follicular layer or not, had a membrane potential of about ‐50 mV in De Boer's solution (DB), but underwent a deep hyper‐polarization of up to ‐90 mV when pretreated with Ca, Mg‐free EDTA‐solution. Regardless of the magnitude of their resting potentials, the defolliculated oocytes exposed to progesterone underwent a gradual depolarization before the germinal vesicle breakdown and retained membrane potential at a level of ‐10 mV until 18 hr post hormone treatment (PHT), the stage of the second meiotic metaphase. A positive‐going activation potential of a magnitude of 70 mV was recorded in the oocytes when pricked at 18 hr PHT as well as in uterine eggs 3–5 min after insemination. A low magnitude of activation potential in response to pricking was recorded in 63% of the oocytes at 13 hr PHT, and premature oocytes exhibiting the activation potential always underwent cortical granule breakdown (CGBD) and perivitelline space formatión. Oocytes where the germinal vesicle had been removed before the hormone treatment exhibited an activation potential and underwent CGBD in response to pricking at 18 hr PHT, whereas those pulse‐treated with cycloheximide (10 μg/ml) during the 8–11 hr PHT exhibited neither of these cortical responses. These results indicate that the syntheses of proteins independent of germinal vesicle taking place at 9–11 hr PHT enable the oocytes to undergo cortical responses.
Lactate Dependent Protein Synthesis in Round Spermatids from Rat Testes
Authors: Nakamura Masahisa , KATO JUNZO
Published: January 2005

Lactate (20 mM) markedly increased protein labeling of round spermatids (steps 1–8) from rat testes. The stimulatory effect of lactate on protein labeling was also observed to some degree in spermatocytes and late spermatids (steps 13–16), but not in Leydig cells and 7 day‐old testis cell suspznsions. In the lactate‐treated spermatids, 51 % of the labeled proteins was found in the water‐soluble fraction (the 105,000 ×g 1 hr supernatant), whereas only 21%, in the control cells. The labeled proteins did not break down for at least 90 minutes in the presence of lactate. Actinomycin D (20 μg/ml) had no effect on [3H]leucine incorporation into the water‐soluble proteins of spermatids, while labeling of the water‐insoluble proteins (the 105,000 ×g 1 hr pellet) was decreased by 23%.

These findings suggest that the round spermatids might be the most susceptible for the lactate‐induced stimulation of protein synthesis among various types of testicular cells, and that lactate increases the synthesis of water‐soluble proteins which may be regulated in the translational level of protein synthesis.
The Degree of Differentiation in Early‐ and Late‐onset Forms of Platyfish‐swordtail Hybrid Melanomas: A Morphological and Physiological Study in Primary Culture
Authors: WAKAMATSU YUKO , OZATO KENJIRO
Published: January 2005

The early‐ and late‐onset forms of platyfish‐swordtail hybrid melanomas (fry and adult melanomas) and the macromelanophores of platyfish and melanotic hybrids were cultured and characterized according to their cellular morphology and physiology. The fry melanomas contained many large and broad cells. The pigmentation of these cells was somewhat less than that of the macromelanophores of platyfish. Most of the fry melanoma cells responded rapidly to 10−6M epinephrine, exhibiting reversible melanosome aggregation. The adult melanomas consisted of small, dendritic, and sparsely pigmented cells. The physiological response of these adult melanoma cells varied widely from tumor to tumor. These findings are discussed in relation to the differentiation of fish melanophores.
Phosphorylation of Deoxyribonucleosides and DNA Synthesis during Embryogenesis of the Sea Urchin
Authors: SUZUKI NORIO , NAGANO HIROSHI
Published: January 2005

The phosphorylation of thymidine, deoxycytidine, deoxyadenosine and deoxyguanosine was studied during the embryogenesis of the sea urchin Hemicentrotus pulcherrimus. [3H]Thymidine was taken up, phosphorylated and accumulated mostly as [3H]thymidine triphosphate in the early cleavage stage embryos. As the embryos developed, the formation of [3H]thymidine triphosphate decreased and most of the [3H]thymidine taken up by the blastulae remained be phosphorylated. When [3H]deoxycytidine was added to the cleaving embryos, the resultant labeled pool consisted of almost equal amounts of [3H]deoxycytidine monophosphate and [3H]deoxycytidine triphosphate. The formation of [3H]deoxycytidine monophosphate increased up to 10 hr following fertilization and then decreased, while the formation of [3H]deoxycytidine triphosphate decreased for 10 hr following fertilization and then gradually increased. [3H]Deoxyadenosine was rapidly phosphorylated to monophosphate derivative in the cleavage stage embryos. The formation of [3H]deoxyadenosine triphosphate increased rapidly after cleavage stage with a concomitant decrease of [3H]deoxyadenosine monophosphate. The activity of phosphorylation in [3H]deoxyguanosine to triphosphate derivative increased rapidly reaching a plateau 10 hr after fertilization. At this point, 80 % of the [3H]deoxyguanosine was recovered as [3H]deoxyguanosine triphosphate.

Based on the above results, it was concluded that the profile of production of each deoxyribonucleoside triphosphate changed during the embryogenesis of the sea urchin, and the in vivo rate‐limiting step of phosphorylation of the individual deoxyribonucleoside was assumed to be different.
Accumulation of Annulate Lamellae in the Subcortical Layer during Progesterone‐induced Oocyte Maturation in Xenopus Laevis
Authors: IMOH HIROSHI , OKAMOTO MITSUMASA , EGUCHI GORO
Published: January 2005

Changes in the fine structure, the location and the number of stacks of annulate lamellae during progesterone‐induced maturation of oocytes of Xenopus were determined by electron microscopy. longitudinal sections of full‐grown oocytes, about 260 stacks of annulate lamellae were observed with marked concentration in the subcortical layer, particularly in the vegetal hemisphere. After exposure to. progesterone, annulate lamellae increased and accumulated further in the subcortical layer. A significant increase of annulate lamellae around the vegetal side of the germinal vesicle seen 3 h after progesterone application. In oocytes 6 h after progesterone application, an average of 320 stacks of annulate lamellae were seen in longitudinal sections and more than two‐thirds of the pore complexes of annulate lamellae were localized in the subcortical layer less than 50 from the oocyte surface, the rest being distributed in the deeper ooplasm. At the time of ger‐ minal vesicle breakdown, all the annulate lamellae underwent complete decomposition. The results were discussed from the view point of comparative developmental biology.
Formation of Embryo Thromboblasts in Chick Blastoderm: Morphology, Site of Production and Time of Emergence in the Blood
Authors: TAHARA YUTAKA , OMORI SHIGEKI , HASHIMOTO AKIHIKO
Published: January 2005

Thrombocytes in the blood of chick embryos (termed embryo thrombocytes by Lucas and Jamroz) have PAS‐positive granules in their cytoplasm. Electron microscopic observations reveal that the embryo thrombocytes contain glycogen granules present singly or in clumps. The presence of these inclusions and other morphological characteristics were used as specific markers to distinguish embryo thrombocytes from primitive erythroid cells. These markers also made it possible to determine the time at which the immature thromboblasts first emerge in blood vessels, and the period of their continued presence in the circulation. In this way we found that thromboblasts were detectable in embryos as early as stage 10+ of Hamburger and Hamilton (after 35 hr incubation) and that the thromboblasts were present in the circulation until day 4 of incubation (stage 23). In ovo and in vitro culture of de‐embryonated blastoderm demonstrated that thromboblasts were formed in the area opeca vasculosa. The present observations suggest that embryo thromboblasts are formed at the same time and in the same area as the primitive cells of erythroid line.
Germline‐specific Antigens Identified by Monoclonal Antibodies in the Nematode Caenorhabditis elegans1
Authors: YAMAGUCHI YASUNORI , MURAKAMI KENJI , FURUSAWA MITSURU , MIWA JOHJI
Published: January 2005

Of 27 monoclonal antibodies identified to react, by indirect immunofluorescent antibody staining, with specific cells and tissues of the nematode Caenorhabditis elegans, we report here three monoclonal antibodies pertaining to the gonadal tissues. One antibody defines an antigen that is distributed over the entire embryo at earlier development and later becomes unique to the gonad, including mature oocytes. The antigens recognized by the other two are distributed asymmetrically in the posterior region of the fertilized egg's cytoplasm destined to become the germline precursor cell. Each antigen is successively segregated only to the germline precursor cells of the developing embryo and, postembryonically, is uniquely localized around the germline cell nuclei of the larvae and adults.
Ribosomal RNA Synthesis in Sea Urchin Embryos: Differential Rates of Accumulation in Chemically‐induced Animalized and Vegetalized Larvae
Authors: O'Melia Anne F.
Published: January 2005

Animalization was induced with evans blue and with Zn++ in embryos of Arbacia punctulata and of Lytechinus variegatus, respectively. Li+ induced vegetalization in A. punctulata embryos. While animalization did not affect the rate of cleavage, vegetalized embryos exhibited a reduction in cell number at post‐morula stages. Mid‐gastrulae and corresponding experimental embryos each were labeled for 4 hr with uridine‐[5‐3H] and with L‐[3H‐methyl]‐methionine. The rate of uptake of each exogenous RNA precursor was similar in control and in experimental embryos. Purified RNA preparations were fractionated by electrophoresis on 2.4% acrylamide+0.5 % agarose gels. Comparison of rates of incorporation of each RNA precursor into 26s and 18s RNAs indicated that on a per cell basis evans blue‐ and Zn++‐animalized embryos showed a reduction (0.53–0.56) and Li+‐vegetalized embryos an enhancement (1.41—1.53) in the rate of accumulation of newly made 26s and 18s RNAs compared to controls (1.00). These results suggest that chemically‐induced animalized and vegetalized embryos provide useful tools for studying possible differential gene expression in different embryonic germ layers of the developing sea urchin embryo.
Effects of a Carcinogen, N‐methyl‐N′‐nitro‐N‐nitrosoguanidine on Blastema Cells and Blastema Formation in Newt Limb Regeneration
Authors: TSONIS PANAGIOTIS A. , EGUCHI GORO
Published: January 2005

The effects of a potent, carcinogen, N‐methyl‐N′‐nitro‐N‐nitrosoguanidine (MNNG) on blastema formation and blastema cells were studied in the adult newt by means of scanning electron microscopy. By administration of MNNG to the blastema of the limbs the formation of the basement lamella, which lies between the epithelium and the mesenchyme, was effectively inhibited at least for a month. Basement lamella in treated limbs is formed 30 or 40 days after carcinogen administration. MNNG altered significantly cell surface morphology and cell motility of blastema cells isolated in vitro. Membrane's specializations such as the formation of filopodia were inhibited. Motility of the treated cells was much reduced compared to the control. The role of these alterations in the carcinogen‐induced abnormal regeneration is discussed.
Evidence for the Presence of a Low Molecular Weight Factor in Sera from Various Animals that Induces in vitro Maturation of Fish Oocytes
Authors: IWAMATSU TAKASHI
Published: January 2005

Studies were made on whether sera from various animals or serum components could induce in vitro maturation of medaka oocytes in the absence of any exogenous hormone. When intra‐follicular oocytes were preincubated in medium containing serum of some mammals or chickens, and were then washed, they matured during further incubation in the same medium without the serum. In contrast, no remarkable stimulatory effect on oocyte maturation was recognized in sera from some other mammals and from reptiles, amphibian and fish. A dialysate of rabbit serum collected using a cellulose membrane (mol. wt. retention 3,500) also induced in vitro maturation of intra‐follicular oocytes. This activity was stable on treatment with proteolytic enzymes or heat‐treatment. Small molecular weight fractions with this activity were obtained by filtering the dialysate through Sephadex G‐10. The oocytes matured by the serum factor were capable of developing normally beyond the stage of embryonic body formation.

From these results, it is concluded that the serum of higher vertebrates contains a low molecular weight factor that stimulates in vitro nuclear and cytoplasmic maturation of intrafollicular fish oocytes.
Induction of Macrocyst Formation by Factors Secreted by Giant Cells in Dictyostelium discoideum
Authors: LAMPHIER MARC S. , YANAGISAWA KAICHIRO
Published: January 2005

The fusion of cells of complementary mating types to produce giant cells has been shown to be the critical event to induce macrocyst formation in Dictyostelium discoideum. We have examined the way in which giant cells use diffusible factors to influence the developmental mode of nearby cells using an experimental design in which NC4 cells are allowed to develop on a dialysis membrane above a suspension of giant cells. We have observed that giant cells are able to inhibit independent aggregation and stream formation in the upper cells and become the dominant aggregation centers. In addition giant cells are able to redirect local amoeba away from the fruiting‐body and toward the macrocyst mode of development. We show that these effects are mediated by diffusible factors of under 2,000 MW. and discuss possible mechanisms of action.
Developmental Change in Microheterogeneity of Serum Transferrin of Chickens
Authors: KIMURA ICHIRO
Published: January 2005

Developmental change in microheterogeneity of chicken serum transferrin (Tf) was investigated by polyacrylsmide‐gel isoelectric focusing, direct immunofixation, and densitometry. Three main Tf species (Tf‐0, Tf‐1, and Tf‐2, which have 0, 1, and 2 sialic acid residues per molecule, respectively) were resolved and their relative ratios were determined. As development proceeded, a relative increase occurred in the most acidic species (Tf‐2) with decreases in the less acidic ones (Tf‐0 and Tf‐1).
Implication of Cytoplasmic Factors in the Lethality of Bufonidae Nucleocytoplasmic Hybrids
Authors: DELARUE MICHEL , AIMAR CHRISTIAN
Published: January 2005

The nucleocytoplasmic hybrid combination performed by nuclear graft between B. bufo nucleus and B. calamita egg is 95% lethal during gastrulation mostly at the young gastrula stage (75%) and to a lesser extent at the yolk plug stage (20%). The heterologous B. calamita cytoplasm was treated or fractioned by different techniques including centrifugation, nucleic acid extraction, ammonium sulfate treatment and heating.

The treated cytoplasmic fractions were injected in B. bufo fertilized eggs in order to check their influence on morphogenetic capacities of the B. bufo nucleus. Injection of hyaline cytoplasmic fraction obtained from B. calamita centrifuged eggs blocks the B. Bufo fertilized egg gastrulation in similar rates as in nucleocytoplasmic association performed by interspecific nuclear grafting. Gastrulation, however, is not affected by injection of total nucleic acids extracted from this hyaline fraction. The soluble cytoplasmic fraction obtained after 60% ammonium sulfate precipitation blocks the development at the beginning of gastrulation, this property being inactivated by heating for 5 min at 60°C. Supernatant from 70% ammonium sulfate precipitation blocks the development at the yolk plug stage and is inactivated by heating for 5 min at 80°C.

These experiments suggest that two distinct factors in B. calamita eggs are involved in developmental arrest of injected B. bufo fertilized eggs, these two factors respectively acting on the beginning and on the end of gastrulation.
Stimulation of Spermatid Phosphofructokinase by Fructose 2,6‐Bisphosphate from Rat Testes
Authors: NAKAMURA MASAHISA , SUZUKI AKIRA , OKINAGA SHOICHI , ARAI KIYOSHI
Published: January 2005

Effect of fructose 2, 6‐bisphosphate on 6‐phosphofructokinase (ATP: D‐fructose‐6‐phosphate 1‐phosphotransferase, EC 2.7.1.11) in spermatid extract from rat testes was studied. Fructose 2, 6‐bisphosphate stimulated the enzyme greatly by increasing its affinity for fructose 6‐phosphate and relieving the inhibition by ATP. Fructose 2, 6‐bisphosphate (0.8 μM) was required for 50% activation of 6‐phosphofructokinase (PFK). In addition, fructose 2, 6‐bisphosphate, AMP and fructose 6‐phosphate acted cooperatively to stimulate the activity of PFK. This stimulation may play an important role in the regulation of glycolysis in spermatids of rat testes.
Effects of D2O on Parthenogenetic Activation and Cleavage in the Sea Urchin Egg*
Authors: KOJIMA MANABU K.
Published: January 2005

When unfertilized sea urchin eggs are treated with D2O‐sea water, fertilization membranes are raised, cytasters appear in the cytoplasm and cleavages occur. Such observations are in agreement with those reported by Gross and his collaborators. However, D2O‐treated eggs can develop at least to the gastrula stage under appropriate conditions, in contradiction to descriptions by Grosset al. This is the case in eggs of three species of Japanese sea urchins used here.

The double treatment with D2O is more effective in inducing parthenogenetic development than the single treatment and, further, double treatments, such as the combinations of D2O and hypertonic sea water or butyric acid and D2O, also, can effectively induce the development.

In addition, centrifugally enucleated egg‐halves can be activated and developed to a blastulalike stage by the double treatment with D2O.
Expression of Toxin Receptors on Cell Surfaces in Transdifferentiating Cultures of Neural Retina
Authors: De POMERAI D. I. , TAKAGI S. , KONDOH H. , OKADA T. S.
Published: January 2005

It has often been asked which of the cell types found during the early stages of culturing embryonic chick neural retina can undergo transdifferentiation into lens in vitro. Since neuronal cell‐surface toxin receptors are maintained in NR cultures for much longer than internal neuronal enzymes (e.g. choline acetyltransferase), and since the transdifferentiation process can be greatly accelerated by preparing reaggregates of neural retina cells after about 10 days of preculture as “monolayers”, a direct test of this question became feasible. 7 or 9 day embryonic chick neural retina cells, precultured for 10–12 days as monolayers, were dissociated and reaggregated under continuous gyration. Reaggregates were maintained for 8 days in the presence of either tetanus toxin or FITC‐conjugated α‐bungarotoxin, to permit surface‐bound toxins to become internalised via receptor turnover. The reaggregates were then dissociated, stained with rabbit antitoxin and FITC‐conjugated anti‐antibody in the case of tetanus toxin‐labelled material, and restained with a rat or mouse antibody against chick δ crystallin followed by the appropriate rhodamine‐conjugated anti‐antibody. Although both FITC/toxin‐labelled cells (putative neurones) and rhodamine/δ crystallin‐labelled cells (transdifferentiated lens cells) were abundant, no examples of double‐labelled cells were observed with 9 day starting material, and only a very few with 7 day starting material. We conclude that the vast majority of differentiated neuronal cells expressing surface receptors for these toxins do not transdifferentiate directly into lens cells.
Control of Late Development of Dictyostelium discoideum by Proteins Functioning at Early Stages
Authors: FUJIKI NAKAO , TAKECHI MAMORU , ISHIKAWA AKIRA
Published: January 2005

We examined two mutants of D. discoideum which are temperature‐sensitive for development. At the nonpermissive temperature one mutant becomes arrested in development during the transition from the finger to the migrating slug. Temperature‐shift experiment indicates that the temperature‐sensitive period begins at considerably earlier tip‐forming stage. The other mutant becomes arrested at the Mexican hat stage and the temperature‐sensitive period coinsided with this stage. The analysis of protein synthesis by two‐dimensional gels, however, showed specific changes at the nonpermissive temperature at an earlier finger‐forming stage.

These results indicate the presence of a control of late development by proteins at early stages.
Appearance and Distribution of RNA‐Rich Cytoplasms in the Embryo of Xenopus laevis during Early Development
Authors: IMOH HIROSHI
Published: January 2005

The occurrence of the dorsal yolk‐free cytoplasm in the fertilized egg of Xenopus was re‐examined, and the appearance and the distribution of RNA‐rich cytoplasms in Xenopus embryos during early development were examined with their paraffin sections. The results show that the dorsal yolk‐free cytoplasm does not occur solely in the dorsal part of the embryo but is continuous to similar cytoplasmic mass in the central and the ventral part. The whole mass of this continuous cytoplasm is denoted here as the mesoplasm. The locations of the mesoplasm in the embryo can be traced by its high RNA content during cleavage and blastulation. The cells endowed with the mesoplasm constitute a broad band about the equator of the blastula. At the lower edge of this band, the blastopore lip is formed during gastrulation. Another mass of yolk‐poor and RNA‐rich cytoplasm becomes distinct around every nucleus in the stage 4 embryo and is denoted here as the nucleophilic plasm. This plasm is diminished at every nuclear division and disappears in the stage 10 embryo. Origins and roles of the mesoplasm and the nucleophilic plasm were discussed and a mechanism of blastulation was suggested.
Developmental Changes in the Activity of Xanthine Dehydrogenase and closely Related Enzyme in Chick Tissues
Authors: IRIE SHINKICHI , KATO YOSHIHIRO
Published: January 2005

Xanthine‐oxidizing activities in the chick tissues were measured by a couple of assay procedures during development of chick embryo. With the usual assay using pterine and NAD+, no detectable level of XDH activity was observed in the liver and little in the duodenum before hatching, whereas an appreciable activity was detected in the kidney of chick embryo. When assayed with xanthine and dichlorophenol indophenol, an XDH‐like enzyme activity was significantly detected in the embryonic liver, while no further enhancement of the activity was detected in the kidney and duodenum. Electrophoregrams obtained with samples from various developmental stages, followed by activity staining with tetrazolium dye, supported the above results and revealed that the embryonic XDH‐like enzyme is not distinguishable from XDH of adult tissue in molecular size. This XDH‐like enzyme, pre‐existing in the liver before hatching, however, exhibited no cross reaction with antibody against intact XDH. The nature of this material was discussed in comparison with deflavinated XDH.
Studies of Unequal Cleavage in Molluscs: I. Nuclear Behavior and Anchorage of a Spindle Pole to Cortex as Revealed by Isolation Technique
Authors: DAN KATSUMA , ITO SUSUMU
Published: January 2005

Unequal division of the eggs of Spisula solidissima was studied by isolating the nuclear elements. The isolation technique consists mainly of the use of Na‐lauryl sulphate with occasional inverting of the container of alcohol‐fixed eggs. The procedure was applied to the following stages. (1) germinal vesicles, (2) 1st maturation division, showing anchorage of a spindle pole to the cortex, (3) 2nd meiosis, (4) pronuclear encouter, (5) metaphase, (6) shifting of the metaphase spindle to one side of the cell and a contact with the cortex, (7) ana‐ and telophase of the 1st division.

Migration and “anchorage” of the spindle to the cortex by one pole is a common phenomenon in unequal divisions of polar body formation and of egg cleavage.
Changes of Chromosome Length and Constitutive Heterochromatin in Association with Cell Division during Early Development of Cynops pyrrhogaster embryo
Authors: YAMAZAKI‐YAMAMOTO KIYOKO , TAKATA KENZO , KATO YOSHIHIRO
Published: January 2005

The length of chromosomes in the presumptive ectoderms of Cynops embryos was measured at nine successive cell divisions from the 6th (cleavage stage) to the 14th (gastrula stage). Up to the 10th cell division (cleavage stage) the chromosome length remained constant. At the 11th cell division the chromosomes began to shorten and continued to shorten at every cell division up to the 14th cell division. The degree of shortening and the mode of variation in length corresponded to the respective developmental stages of cleavage, blastula and gastrula. During those periods when chromosomes became shortened, some fine C‐bands of the paracentromeric region found in earlier stages fused together. The chromatin of interphase nuclei also showed considerable changes during chromosome shortening. Besides the size reduction of interphase nuclei, the number of C‐band granules in an interphase nucleus decreased in parallel with chromosome shortening and fusion of C‐bands in mitotic chromosome.
Anion Channel Blockers Inhibit the Acrosome Reaction of Echinoderm Sperm
Authors: NISHIYAMA ICHIRO , SASAKI HAJIME , MATSUI TAEI , HOSHI MOTONORI
Published: January 2005

Two types of anion channel blockers, SITS (4‐acetamide‐4‘‐isothiocyanostilbene‐2,2′‐disulfonic acid) and DIDS (4,4′‐diisothiocyanostilbene‐2, 2’‐disulfonic acid), inhibited jelly‐induced acrosorne reaction in starfish and sea urchin. In starfish sperm, both of the blockers reversibly inhibited the formation of acrosomal process but they had no effect on either the acrosomal exocytosis or acid release from the sperm. Complete acrosome reaction occurred even in Cl−‐ and SO42−‐free artificial seawater whereas HCO3−was required for the acrosomal exocytosis. Importance of anion transport in acrosome reaction is discussed.
Acceleration of the Cleavage in Sea Urchin Eggs by Treatmentswith Local Anesthetics (I) Procaine Treatment*
Authors: KOJIMA MANABU K.
Published: January 2005

Unfertilized sea urchin eggs were pre‐treated with 2–10 mM procaine sea water for 10 min and then fertilized 5 min later. These eggs hatched about 1–1.5 hr earlier than the control eggs. This acceleration of hatching seems to be the result of a reduction in the individual cleavage cycles.

In the second series of experiments, eggs were inseminated at 5, 30 and 60 min after the procaine treatments. In this case, the rate of accelerated cleavage decreased gradually, as the intervals between the treatments and insemination were extended.

Next, the minimum time‐length of procaine treatment to accelerate cleavage was determined. When eggs were immersed in procaine for 1 min, cleavage was accelerated as in the case of 10 min‐treatment, although acceleration to the same degree required higher concentrations of procaine solution.

When fertilized eggs were post‐treated with procaine, cleavage was also accelerated, but both the rate of acceleration and the effective concentrations decreased with time after fertilization.
Continuous Cell Lines from Larval Hemocytes of the Cabbage Armyworm, Mamestra brassiae
Authors: MITSUHASHI JUN , SHOZAWA AKIKO
Published: January 2005

Two continuous cell lines, NIAS‐MaBr‐92 and NIAS‐MaBr‐93, were obtained from larval hemocytes of the cabbage armyworm, Mamestra brassicae. The cells grew in suspended state. Spherical cells were predominant, although there were spindle shaped and irregular shaped cells. The chromosome numbers of the cell lines varied very much with the mode of 100 of 120. The population doubling times of these cell lines were about 36hr. The cells could grow in media free of serum and lacking sterols, fatty acids and protein. The cell lines consumed ammonia and produced glutamine when cultured in MM medium. The cells could be stored at ‐100°C for more than two year, and at 5°C for two months. The cells were sensitive to Autographa californica nuclear polyhedrosis virus and Chilo iridescent virus.
Detection of the Initial Step of Mesenchymal Differentiation of Teratocarcinoma Cells Using the Monoclonal Antibody Eccd‐11
Authors: YOSHIDA‐NORO CHIKAKO , TAKEICHI MASATOSHI , OKADA T. S.
Published: January 2005

The monoclonal antibody ECCD‐1 recognizes the Ca2+‐dependent cell‐cell adhesion molecule of teratocarcinoma stem cells (EC cells) and of a certain class of differentiated epithelial cells. It actively disrupts cell‐cell adhesion when added to monolayer cultures of these cells, but does not affect adhesion of mesenchymal or neuronal cells. When ECCD‐1 was added to clonal cultures of EC cells (PCC3/A/1 line), all the cells were initially sensitive to the antibody, but after 5 to 6 days of culture a fraction of the cells in certain colonies no longer reacted with the antibody although they expressed alkaline phosphatase activity, which is a marker of undifferentiated EC cells. We isolated these ECCD‐1‐resistant cells by recloning and examined their differentiation by clonal culture. Most of them differentiated into fibroblastic cells and a few into skeletal muscle‐like cells, but none differentiated into any other cell types. From these observations, we suppose that the ECCD‐1‐resistant population of EC cells are committed to mesenchymal differentiation. The use of ECCD‐1, thus, permitted us to detect EC cells at the initial stage of a particular differentiation pathway.
Steroid Producing Cells during Ovarian Differentiation of the Tilapia, Sarotherodon niloticus
Authors: NAKAMURA MASARU , NAGAHAMA YOSHITAKA
Published: January 2005

In order to examine the initial appearance and development of the steroid producing cells (SPCs) during the process of ovarian differentiation, histology and ultrastructure of tilapia (Sarotherodon niloticus) ovaries were investigated from 10 to 50 days after hatching. In gonads of fry at 23–26 days after hatching, initial ovarian differentiation was confirmed by the differentiation of stromal aggregations in the proximal and distal region of the gonad on the side facing the lateral wall. This represents the initial formation of the ovarian cavity. At the same time as ovarian differentiation, a few large cells appeared initially in the vicinity of blood vessels. They have some of the ultrastructural features characteristic of SPCs such as a moderate number of mitochondria with tubular cristae, a large amount of smooth endoplasmic reticulum and many free ribosomes. Based on these ultrastructural criteria, together with the present finding that these cells further differentiated into the typical SPCs at older stages, these cells were identified as SPCs. Thereafter, by 30–50 days, SPCs increased gradually in number in the area enclosing the blood vessels of ovaries. The increase in SPCs coincided with the development of germ cells, including the multiplication of oogonia and the transformation from oogonia to oocytes.
The Effect of Hydrocortisone on the Epidermal Proliferation in an Organ Culture of Chick Embryonic Skin
Authors: SUZUKI‐MORIMOTO SHIGEKO , YAMAMOTO YOSHIHIRO , YAMAGUCHI TAKEO
Published: January 2005

Hydrocortisone is regarded as an initiator of keratinization in embryonic skin. The present investigation dealt with the effect of hydrocortisone on the proliferation of epidermal cells during early development: Cell kinetic analyses using 3H‐thymidine autoradiography were applied to a skin organ culture prepared from a 13‐day chick embryo.

Hydrocortisone at a concentration between 0.01 and 1.0 μg/ml was effective in initiating a morphological change leading to the epidermal keratinization in vitro and caused a marked decrease in the mitotic and labeling indices of epidermal basal cells, the decrease being maximum at 2 days of culture previous to the morphological change.

During continuous labeling with 3H‐thymidine, the number of labeled basal cells reached 100% within 2 days in the control and 4 days in the culture treated with hydrocortisone. This confirmed that the growth fraction of epidermal basal cells was 1.0 even after the administration of hydrocortisone.

The duration of each cell cycle phase at 2 days of culture was determined by percent labeled mitoses and double‐labeling analyses. It was concluded that hydrocortisone extended the generation time of epidermal basal cells at this time point about three fold over the control. This extension was mainly due to the elongation of the G1 phase.