PubMed Health⌕ Search

Biomedical subjects

N Wakasugi

Publications and source records attributed to N Wakasugi.

At least 37 records · Page 2Linked to original sources
PubMed1991

Stumpy limb--an embryonic lethal mutation in Japanese quail (Coturnix coturnix japonica).

An embryonic lethal mutant was found in Japanese quail and named "stumpy limb (SL)". All SL embryos died at pre-hatching stages, showing brachycephaly, a thickened neck, short upper and lower beaks, and short and thick extremities, while their body length was similar to that of the normal embryos. Observations on the skeleton revealed a globular skull, unusual curvature of the Processus palatinus maxillaris of the upper beak, and shortening and thickening of the appendicular bones. Some embryos showed a bending of the humerus, femur and/or tibiotarsus. Abnormality was more conspicuous in the leg bones than in the wing bones. No conspicuous differences were observed in the vertebrae between the SL and normal embryos. A genetic analysis suggested that the mutation is controlled by an autosomal recessive gene, for which the gene symbol sl was proposed.

Animals↗
PubMed1990

Role of ATL-derived factor (ADF) in the normal and abnormal cellular activation: involvement of dithiol related reduction.

HTLV-I transformed T cells not only express a large number of interleukin-2 receptors (IL-2R/p55(Tac], but also produce an IL-2R/Tac inducer named ATL-derived factor (ADF). We have cloned the ADF cDNA and found that ADF production in human lymphocytes can be enhanced by cellular activators such as mitogens or phorbol esters. Recombinant ADF produced by E. coli was shown to have growth-promoting activity in combination with interleukin-2 or suboptimal mitogenic stimuli on several lymphoid cells including human PBMCs, besides the originally reported IL-2R/Tac inducing activity. Homology analysis revealed an unexpected structural relationship between ADF and dithiol-reducing enzyme, thioredoxin, which had been characterized originally in prokaryotic system. Recombinant ADF also has a reducing activity, suggesting the presence of still unknown features of ADF action in vivo. The requirement of dithiol reduction in the biological activities of ADF, together with the possible involvement of ADF production in the normal and abnormal activation of human cells are discussed.

Amino Acid Sequence↗
PubMed1990

Adult T-cell leukemia-derived factor/thioredoxin, produced by both human T-lymphotropic virus type I- and Epstein-Barr virus-transformed lymphocytes, acts as an autocrine growth factor and synergizes with interleukin 1 and interleukin 2.

Interleukin 1 (IL-1) has been obtained from the Epstein-Barr virus-infected B-lymphoblastoid cell line 3B6 and shown to be involved in autocrine growth of 3B6 B cells. Independently, adult T-cell leukemia-derived factor (ADF) was purified from human T-lymphotropic virus I-infected leukemic T-cell line (ATL-2) and reported as an interleukin 2 (IL-2) receptor-inducing factor. We have previously reported the same molecular mass, pI, and NH2-terminal amino acid sequence for both 3B6-derived IL-1 and ADF. cDNA cloning of ADF demonstrated high homology with the prokaryotic disulfide reducing enzyme thioredoxin. We show here that ADF and 3B6-derived IL-1 are identical. By RNA blot, 3B6 and ATL-2 cells were shown to contain high levels of 0.6-kilobase mRNA corresponding to ADF. Such message was not detected in resting peripheral blood lymphocytes but could be weakly induced by lymphocyte activation. Antibodies have been raised against synthetic peptides corresponding to the NH2 terminus and the COOH terminus of ADF. Immunoblotting and sequential immunoprecipitation with these antibodies revealed the same 13-kDa protein in 3B6 and ATL-2 cells. Recombinant ADF could sustain growth of 3B6 and ATL-2 cells at low cellular concentration without fetal calf serum; ADF, thus, appears involved in their autocrine growth. Similarly, recombinant ADF could enhance growth of other B-cell lines, including the Epstein-Barr virus-negative Burkitt lymphoma line BL41 and the lymphoblastoid cell lines CRAG8, CRB95, and 1G8. Finally, recombinant ADF exhibits marked synergism with other cytokines, such as IL-1 and IL-2, allowing virally infected lymphocytes to respond to suboptimal amounts of a variety of growth factors.

B-Lymphocytes↗
PubMed1990

Head skeletal abnormalities associated with the throat tuft mutation (hfdTt) in the Japanese quail.

Hereditary abnormalities of head skeleton ascribed to embryonic visceral arch defects were investigated in Japanese quail homozygous for the throat tuft mutation (hfdTt). Differential staining for bone and cartilage of 15-day embryos revealed partial deletion or irregularity in the mandible, basiparasphenoid, hyoid apparatus, and quadratojugal bones, incidences of which were 84, 44, 40, and 18%, respectively. Incidence of the head-skeleton abnormality was 88% when individuals having at least one of the abnormalities mentioned were regarded as abnormal. Throat tuft (hfdTt) and ear tuft (hfd) homozygotes showed distinct differences between them, although both alleles are associated with visceral arch defects.

Animals↗
PubMed1989

Two distinct affinity binding sites for IL-1 on human cell lines.

We used two human cell lines, NK-like YT-C3 and an EBV-containing B cell line, 3B6, as models to study the receptor(s) for IL-1. Two distinct types of saturable binding sites were found on both cell lines at 37 degrees C. Between 1 pM and 100 pM of 125I-IL-1-alpha concentration, saturable binding sites were detected on the YT-C3 cells with a K of 4 x 10(-11) M. The K found for the IL-1-alpha binding sites on 3B6 cells was 7.5 x 10(-11) M. An additional binding curve was detected above 100 pM on YT-C3 cells with a K of 7 x 10(-9) M and on 3B6 cells with a K of 5 x 10(-9) M. Scatchard plot analysis revealed 600 sites/cell with high affinity binding and 7000 sites/cell with low affinity for YT-C3 cells and 300 sites/cell with high affinity binding and 6000 sites/cell with low affinity for 3B6 cells. At 37 degrees C, the internalization of 125I-labeled IL-1 occurred via both high and low affinity IL-1R on both YT-C3 and 3B6 cells, whereas the rates of internalization for high affinity binding sites on YT-C3 cells were predominant in comparison to that of low affinity binding sites. In chemical cross-linking studies of 125I-IL-1-alpha to 3B6 and YT-C3 cells, two protein bands were immunoprecipitated with Mr around 85 to 90 kDa leading to an estimation of the Mr of the IL-1R around 68 to 72 kDa. In similar experiments, the Mr found for the IL-1R expressed on the murine T cell line EL4 was slightly higher (around 80 kDa). Whether these distinct affinity binding sites are shared by a single molecule or by various chains remains to be elucidated.

Adolescent↗
PubMed1989

A monoclonal antibody recognizing 68- to 75-kilodalton protein(s) associated with the human IL-1 receptor.

We produced an IgM mAb termed 4.9 against an EBV-containing lymphoblastoid cell line, termed 3B6. This mAb reacted with both various B and T cell lines such as HSB2 cells, with an NK-like cell line YT-C3 cells, and with human fibroblast MCR-5 cells. It also reacted with normal resting peripheral B lymphocytes, monocytes, and anti-CD2- or anti-CD3-activated T lymphocytes. The 4.9 mAb immunoprecipitated two bands estimated to be of Mr 68 and 75 kDa from iodinated 3B6 cells. The 4.9 mAb inhibited the proliferation of peripheral T lymphocytes induced either by anti-CD3 mAb or anti-CD2 mAb. The 4.9 mAb inhibited also the proliferation of murine thymocytes both in the presence of PHA and IL-1 and the proliferation of human fibroblasts in the presence of IL-1. Radiolabeled IL-1 binding on 3B6 cells revealed two types of IL-1 binding sites with high and low affinity for IL-1 (300 sites/cell with a Kd of 6 x 10(-11)M and 6000 sites/cell with a Kd of 3 x 10(-9)M). On both 3B6 and YT-C3 cells, mAb 4.9 inhibited specifically the binding of 125I-labeled rIL-1, alpha or beta, whereas the irrelevant IgM mAb did not. Conversely, rIL-1, alpha or beta, could inhibit specifically the binding of radioiodinated 4.9 mAb to 3B6 or YT-C3 cells, whereas rIL-2, rIFN, or the irrelevant IgM mAb were ineffective. 125I-4.9 mAb bound 3B6 cells with an association constant (Ka) of 2 x 10(8)/M and demonstrated 6000 binding sites/cell. We thus conclude that mAb 4.9 recognizes a protein complex (68 to 75 kDa) closely associated with the IL-1R.

Adult↗
PubMed1988

Comparison of interleukin 2 and 12-O-tetradecanoylphorbol 13-acetate as signals for protein kinase C activation in purified human T lymphocytes.

Interleukin 2 (IL2) and 12-O-tetradecanoylphorbol 13-acetate (TPA) have been compared for their ability to induce translocation of protein kinase C (PKC) in T lymphocytes prestimulated with anti-CD3 monoclonal antibody (mAb), either in the presence or absence of monocytes. TPA alone did not promote purified T cell growth, but it was able to induce a transient, within 30 min, translocation of PKC activity. The profiles of PKC association with the membrane of the T cells under TPA stimulation were quite similar when either the anti-CD3 mAb or the fixed monocytes, or both, were added to the T cells. The decrease of cytosolic PKC under TPA stimulation was less pronounced for the purified T cells stimulated with anti-CD3 mAb, fixed monocytes alone or both than for unstimulated purified T cells. Even in the absence of monocytes, the addition of exogenous IL2 to the anti-CD3 mAb-treated T cells resulted in PKC translocation, with a transient increase in the PKC activity found in both the particulate and cytosolic fractions. When exogenous IL2 was added to the proliferating T cells, the association of PKC with the membrane was prolonged and the activity did not reach a plateau during the first 2 h after the IL2 stimulation. In parallel, the level of PKC associated with the membrane was higher in proliferating cells than in resting cells even 4 days after stimulation. These results suggest that activation of PKC by IL2 might be different from the direct activation of PKC by TPA and that a specific activation pathway, at least kinetically distinct from the classical phosphatidyl inositol diphosphate degradation by phospholipase C, might be involved during IL2 stimulation of T lymphocytes through high-affinity IL2 receptors.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1988

Back-drawer: a behavioral mutant in Japanese quail.

The behavioral mutant "back-drawer" of Japanese quail is characterized by crouching and back-drawing action with the neck bending ventrally and occasional forward rolling. The abnormal behavior appears from hatching to 8 weeks of age. Back-drawers were classified into three types: (1) those showing the abnormality at hatching and dies within 5 days; (2) those showing the abnormality after 2 weeks of age and dies shortly thereafter; and (3) long survivors suffering from abnormal behavior, and showing gradual recovery. Back-drawer males are infertile when showing the abnormality, but can produce progeny after recovery. In contrast, the females tend to have a short life span and lack reproductive ability. Genetic analyses have indicated that this mutant character is controlled by 2 pairs of autosomal recessive genes.

Animals↗
PubMed1988

Reduced fertility in gracile axonal dystrophy (gad) mice.

Mean litter size in gad/gad females was significantly lower than in normal females (+/+ and gad/+) in intra- and inter-strain crosses. The reduction in litter size was not dependent on the genotypes of the males, but could be attributed to the gad/gad females themselves. The numbers of corpora lutea and implants in gad/gad females were slightly reduced as compared with those in the controls, but the number of live fetuses was significantly lower than that in normal females 14 days after copulation (P less than 0.02). Hence, reduced litter size in gad/gad females was accounted for mostly by embryonic and fetal death after implantation, which was inferred to be due to impaired uterine function.

Animals↗
PubMed1988

Gracile axonal dystrophy (GAD), a new neurological mutant in the mouse.

A new neurological mutant has been found in the F2 offspring of CBA/Nga and RFM/Nga mice. Affected mice exhibited ataxia beginning at about 80 days of age, followed by tremor, difficulty in moving, and muscular atrophy of the hind limbs. The neurological signs became progressively severe, and death occurred by 5 to 6 months of age. Since the animals could be distinguished from normal mice by the abnormal positions of the hind limbs when the mouse was hung by the tail after 1 month of age, they could be bred until onset of the signs. Pathological examination revealed neuroaxonal dystrophy and degeneration in the gracile nucleus of the medulla oblongata and the gracile fascicules of the spinal cord, which could be the main cause of the clinical signs. The mutation is inherited as an autosomal recessive trait. It was, therefore, named gracile axonal dystrophy (GAD) with the gene symbol gad. The mice could be a new pathological model for the study of neuroaxonal dystrophy.

Animals↗
PubMed1988

Fertility in intersubspecific hybrids of laboratory mice (Mus musculus domesticus) and molossinus mice (M. m. molossinus).

We studied the reproductive performance of F1 and F2 hybrids of laboratory mice (C57BL/6, B6 and BALB/c) and molossinus mice (MOM and Mol-A). The F1 x F1 crosses were fully fertile. In the F2 x F2 crosses, the copulation rate was slightly lower and the pregnancy rate was markedly depressed: only 5 out of 18 copulated females (27.8%) became pregnant in the F2 hybrids derived from the reciprocal crosses of B6 x MOM, and in the F2 hybrids from BALB/c x Mol-A crosses, the pregnancy rate was 51.4% (18/35). This low fertility was attributed mainly to the F2 females, because there was a much lower pregnancy rate (56.5%; 26/46) in the (B6 x MOM)F2 female x B6 male crosses compared with the B6 female x (B6 x MOM)F2 male crosses (80.6%; 26/32). On the other hand, the pregnant F2 females were judged to have normal reproductive ability, based on observations of the numbers of corpora lutea, implantations and live fetuses at day 14 of pregnancy. Apparently there is segregation of fertile and sterile females at the F2 generation, but it remains to be determined how the loss of fertility is brought about in the sterile F2 females.

Animals↗
PubMed1987

Cryopreservation of strains and mutant genes in mice.

Three kinds of freezing methods were tested with embryos of DNI strain. The survival rate after thawing was 47.5%, 66.7% and 77.8% in the 2-step method, modified slow freezing method and modified 2-step method, respectively. Then, the modified 2-step method was applied to the embryos from 7 strains and a pair of interstrain crosses. PMSG treatment at the beginning of diestrus following HCG treatment after 48 hrs resulted in much yield of 8-16-cell embryos in all strains. The average number for each strain was as follows: DNI; 18.9, DDN; 13.0, BS; 20.4, C57BL/6; 12.9, DBA/2; 17.5, CRN; 19.8, PAN; 13.7 and DNI x C57BL/6-Ay; 21.7. Development of frozen-thawed embryos in culture varied among strains. Proportion of embryos that developed to the morula or blastocyst stage was as follows: DNI; 64.6%, DDN; 71.9%, BS; 53.6%, C57BL/6; 57.3%, DBA/2; 65.0%, CRN; 52.5%, PAN; 17.4% and DNI x C57BL/6-Ay; 44.1%. These results indicate that the ability of embryos to survive freezing and thawing is influenced by their genetic background. Live young were produced from DNI, DDN, BS and DNI x C57BL/6-Ay embryos after transfer to recipients. Comparative assessment of the developmental ability of frozen-thawed embryos after transfer among strains should be performed in further study.

Animals↗
PubMed1985

Defective IFN-gamma production in the human neonate. II. Role of increased sensitivity to the suppressive effects of prostaglandin E.

Analysis of endogenous production and effects of exogenous addition of interleukin 2 (IL 2), leukotrienes (LT), and prostaglandin E (PGE) has been used to investigate the dysregulation responsible for impaired PHA-induced IFN-gamma secretion by cord blood leukocytes (CBL). The addition of LT or IL 2 could not reverse the IFN defect of CBL. The production of these two mediators was found to be normal in CBL cultures. CBL and control leukocytes from adult donors produced comparable amounts of PGE2. In contrast, sensitivity to the suppressive effects of PGE2 on IFN-gamma secretion was much higher with CBL than with control leukocytes. Treatment with indomethacin reversed the IFN-gamma defect with most CBL tested, and the addition of physiologic amounts of PGE2 to indomethacin-treated cultures resulted in a profound impairment of IFN-gamma production similar to that of untreated CBL cultures. Preincubation of CBL for 24 hr before PHA stimulation resulted in restoration of a normal sensitivity to exogenous PGE2, in parallel with correction of the IFN-gamma defect. Our observations suggest that the impairment of IFN-gamma secretion in neonates is not due to deficient amplification circuits, but is the consequence of an exaggerated cellular sensitivity to the suppressive effects of PGE produced endogenously in normal amounts.

Adult↗
PubMed1985

Defective IFN-gamma production in the human neonate. I. Dysregulation rather than intrinsic abnormality.

Cord blood leukocytes (CBL) stimulated with PHA, Con A, or with the monoclonal antibody OKT3 proliferate normally but produce very low titers of IFN-gamma. This defect was not observed with maternal leukocytes collected at the time of delivery, indicating that the defective production of IFN-gamma in CBL is not a mere consequence of a hormonal change associated with labor. CBL produced large amounts of IFN-gamma (comparable to those observed in adult control and in mothers) after stimulation with staphylococcal enterotoxin A (SEA). Furthermore, gamma-irradiation with as little as 500 or 1000 rad, or incubation at 37 degrees C for 24 hr, reversed the defect in PHA-induced IFN-gamma secretion. This finding indicates that the defective secretion of IFN-gamma of CBL is not intrinsic, but rather is the consequence of a subtle dysregulation. We could not find evidence for a defective accessory function with cord blood monocytes, because the addition of adherent cells from adult donors did not reverse the defect. In co-cultures of adult leukocytes and CBL, PHA-induced IFN-gamma secretion was comparable to that of adult cultures tested alone. Nonirradiated CBL were not able to suppress IFN-gamma secretion by irradiated autologous leukocytes. Together, our results suggest that the defective PHA-induced IFN-gamma secretion of CBL is the result of an original type of dysregulation and is associated with an excessive sensitivity to suppressive signals rather than excessive suppressor function.

Adult↗
PubMed1984

Mineral content of quail embryos cultured in mineral-rich and mineral-free conditions.

Japanese quail embryos were cultured in mineral rich (MR) using chickens egg shell and mineral-free (MF) using Saran Wrap cultures after 2.5 days of normal incubation. In the MR culture, 82% of embryos survived beyond 14.5 days of incubation and 3 embryos out of 93 (3%) hatched. In the MF culture, 80% survived beyond 9.5 days and 59% beyond 14.5 days. Mineral content (Ca, Mg, K, and Na) of the embryos and of the residual matter within the egg (remaining egg contents excluding the embryo, shell, and shell membrane) were analyzed at 7 to 15 days of incubation. Calcium content of the embryos increased gradually between 9 and 11 days and showed a rapid increase after 11 days in both normal incubation (control) and MR culture. Changes in mineral levels of the embryos plus residual matter within the egg were as follows. Calcium content of the controls increased gradually between 9 and 11 days and showed a rapid increase after 11 days. In MR cultures, Ca increased between 10 and 12 days and showed a rapid increase after 12 days. Magnesium content increased rapidly after 13 days in controls and showed a small increase after 13 days in MR cultures. Potassium and Na content remained constant in both controls and MR cultures. In MF cultures, content of the four minerals remained constant throughout the culture period. Mineral levels of Japanese quail and chicken eggs were compared before and after normal incubation. It was found that 81.5% of Ca and 30.8% of Mg assimilated by newly hatched quail chicks were derived from the egg shell during incubation and 84.2% of Ca and 23.5% of Mg in the chicken.

Animals↗
PubMed1983

Development of cultured quail embryos.

Japanese quail embryos were cultured by a plastic wrap technique after 2.5 days of normal incubation, and their development was investigated. Mean survival time was 13 days (2.5 days of preincubation plus 10.5 days of experimental culture). Mean morphological stage shown by the cultured embryos was Stage 28 of Zacchei (1961) (corresponding to 11 days of normal incubation) and the maximum stage of development was Stage 31 (corresponding to 14 days of normal incubation). In the latter half of incubation periods, cultured embryos showed retarded development, and their size was smaller than intact control embryos. Because continual observation of embryos is possible, this culture technique would be useful for experimental manipulation of the embryos or studies on lethal genes.

Animals↗