Osteological development of the feeding apparatus in early stage larvae of the seabass,Lates calcarifer

The osteological development of the feeding apparatus was examined in early stage larvae of laboratory-reared seabass,Lates calcarifer. At initial mouth openings 40 hours after hatching, the larvae were equipped with the fundamental elements forming the oral cavity, such as the trabecular roof, the lower branchial and hyoid arches forming the floor, the quadrate and symplectic-hyomandibular cartilages making up the sides, and the maxilla and Meckel's cartilage bordering the jaws. The cleithrum appeared almost simultaneously. The mechanics of creating a negative pressure in the oral cavity, which results in a “sucking” mode of feeding, were elucidated from these elements. During a period from 50–60 to 100–110 hours after initial mouth opening (HAMO), new elements such as the premaxilla and jaw teeth appeared, and the ossification of existing elements started. The new elements apparently enabled the larvae to “grasp” food organisms, in addition to the already existing and increasing sucking ability, from 100–110 HAMO.     

Plasma lysosomal enzymes after liver transplantation in the pig.

During liver transplantation in the pig, the plasma activities of beta-galactosidase, beta-glucuronidase and beta-glucosidase were elevated as early as 15 min after establishing the hepatic circulation. The enzyme activities peaked at 3 h and returned to the initial level within 2-3 days. However, such substantial alterations were not observed in other enzymes, alpha-mannosidase and alpha-glucosidase. Similar reactions to those of the first three enzymes were found in aspartate aminotransferase and lactate dehydrogenase but with later peaks and slower eliminations. In light of the current study, the serial estimation of acid hydrolases may be useful to discover the extent of tissue injury and also to evaluate the effectiveness of various organ-preservation methods.     

Application of diffractometry and a linear image sensor to measurement of erythrocyte deformability.

We applied laser diffractometry and a linear image sensor to measurement of erythrocyte deformability to detect the light intensity pattern of the diffraction image. Deformability was evaluated as the deformability index (DI), calculated from the width and length of the diffraction pattern ellipse, estimated by the linear image sensor. With the erythrocytes under various shear stresses, the DI was linearly related to results by the geometric method (r = 0.996, p < 0.01). The coefficient of variance of DI at a shear stress of 236 dynes/cm2 was 0.2% (seven human blood samples), which was satisfactory for practical use. The DI was independent of the erythrocyte concentration in the range of 1.5 x 10(7)-5.0 x 10(7) cells/ml of suspension. Correlation between the DI and the logarithm of shear stress was linear in the range of 5 to 350 dynes/cm2 of shear stress in suspension media of different viscosities. Heat-treatment, which decreased membrane flexibility, caused parallel reduction of the DI plotted against the logarithm of shear stress. The method was sensitive and gave reproducible results. It may be useful for clinical applications.     

A highly sensitive enzyme immunoassay of anti-insulin antibodies in guinea pig serum

A highly sensitive enzyme immunoassay of anti-insulin antibodies in guinea pig serum is described. Guinea pig anti-insulin serum was diluted to various extents with nonspecific guinea pig serum and incubated with insulin. After incubation, free insulin was separated from insulin-anti-insulin antibody complex by treatment with dextran-charcoal. Anti-insulin antibodies in the complex were dissociated from insulin by incubation with 0.23 M HCl and inactivated. The amount of dissociated insulin was measured by sandwich enzyme immunoassay using anti-insulin IgG-coated polystyrene balls and affinity-purified anti-insulin Fab'-horseradish peroxidase conjugate. The detection limit of anti-insulin antibodies in guinea pig serum was 6.7 pg/assay or 150 ng/liter of serum. The present enzyme immunoassay was 10,000-fold more sensitive than the previously described enzyme immunoassay, in which insulin-coated polystyrene balls were incubated with diluted guinea pig anti-insulin serum and subsequently with rabbit (anti-guinea pig IgG) Fab'-horseradish peroxidase conjugate.     

Activated lymphocytes in patients with newly diagnosed type 1 (insulin-dependent) diabetes mellitus

The expression of activation antigens (transferrin receptor, IL-2 receptor and Ia antigen) on circulating T lymphocytes from Japanese children with Type 1 diabetes was studied using five monoclonal antibodies (Ab), OKT9, anti-Tac Ab, OKIa1, anti-human HLA-DR Ab and OKT3. For detecting Ia positive T cells, the dual staining technique using OKT3 and anti-Ia antibody was employed. Four out of six patients (67%) with newly diagnosed Type 1 diabetes showed a raised level of either OKT9 or Tac positive cells when examined at diagnosis. These patients, however, rapidly lost these activation antigens after the insulin therapy was started. In contrast, in 32 long-standing patients, only 2 (6%) had a high percentage of OKT9 positive cells and none of them demonstrated Tac positive cells. One out of six newly diagnosed patients or three out of 21 long-standing patients had a significantly high percentage of Ia-positive T cells compared with normal subjects. In poorly controlled long-standing patients whose HbA1 value was higher than 14%, none of them had an increased number of activated lymphocytes. Therefore, it is unlikely that insulin deficiency and hyperglycemia were responsible for the changes observed in these studies. Activated lymphocytes might be related to activation of the immune system involved in pathogenesis of Type 1 diabetes.     

Novobiocin inhibits the SV40 enhancer activity

Using a transient expression assay, we have analysed the effect of novobiocin, DNA topoisomerase II inhibitor, on simian virus 40(SV40) enhancer activities. We used the recombinant clones containing type I or II collagen promoters placed upstream of the bacterial chloramphenicol acetyltransferase (CAT) gene with or without SV40 enhancer. We observed the expected increase in CAT activities due to the presence of the SV40 enhancer. Interestingly, CAT gene expression of the enhancer-containing constructs were inhibited more sensitively by novobiocin than that of the enhancer-less construct. This findings lead us propose that DNA superhelicity mediated by topoisomeraseII is one of the important factor for the manifestation of SV40 enhancer activity.     

Cope's rule and the evolution of body size in Pinnipedimorpha (Mammalia: Carnivora)

Cope's rule describes the evolutionary trend for animal lineages to increase in body size over time. In this study, we tested the validity of Cope's rule for a marine mammal clade, the Pinnipedimorpha, which includes the extinct Desmatophocidae, and extant Phocidae (earless seals), Otariidae (fur seals and sea lions), and Odobenidae (walruses). We tested for the presence of Cope's rule by compiling a large dataset of body size data for extant and fossil pinnipeds and then examined how body size evolved through time. We found that there was a positive relationship between geologic age and body size. However, this trend is the result of differences between early assemblages of small-bodied pinnipeds (Oligocene to early Miocene) and later assemblages (middle Miocene to Pliocene) for which species exhibited greater size diversity. No significant differences were found between the number of increases or decreases in body size within Pinnipedimorpha or within specific pinniped clades. This suggests that the pinniped body size increase was driven by passive diversification into vacant niche space, with the common ancestor of Pinnipedimorpha occurring near the minimum adult body size possible for a marine mammal. Based upon the above results, the evolutionary history of pinnipeds does not follow Cope's rule.     

Amino acid substitution in the C-terminal arm domain of HU-2 results in an enhanced affinity for DNA.

Three mutants of the Escherichia coli hupA gene, encoding the HU-2 protein, were constructed by synthetic oligodeoxyribonucleotide-directed, site-specific mutagenesis on M13mp18 vectors. The resulting HupAN10, HupAN11 and HupAN12 proteins contained Thr59-->Lys, Gln64-->Lys and Asn53-->Arg substitutions, respectively. These amino acid (aa) changes increased the positive charge of the N-terminal half of the two-strand, antiparallel beta-ribbon of the arm structure, which is believed to be a domain for DNA binding. The three mutant proteins bound to DNA more tightly than wild-type HU-2, and their affinities for DNA increased in the order of HupAN10, HupAN11, HupAN12. The mutant proteins showed a slightly increased HU activity for supporting Mu phage development. A mutant HU-2 protein with increased basicity, but with an altered aa sequence in the arm region due to a frameshift mutation, was also constructed. This mutant protein showed a reduced affinity to DNA and was unable to support Mu growth, suggesting that a unique aa sequence of the arm domain, rather than mere basicity of this domain, is required for efficient binding to DNA.     

Chemical synthesis of capped RNA fragments and their ability to complex with eukaryotic ribosomes

In other to examine the binding ability of the 5'-terminal part of eukaryotic mRNA to 80S ribosome, several kinds of oligoribonucleotides, pA-U-G, m7G5'pppA-U-G, m7G5'pppG-U, m7G5'pppA-U-G-A-C-C, were synthesized chemically. The binding experiments of oligonucleotides to 80S ribosome showed that the capped structure as well as AUG are essential for ribosome binding, and the efficiency is enhanced by the 5'-leader sequence if it would include complementary sequence to the 3'-terminal part of 18S rRNA.     

Improving noise resistance of intrinsic rhythms in a square-wave burster model

The square-wave burster (Wang and Rinzel, 2003) is a class of autonomous bursting cells that share a bifurcation structure. It is known that this class of cells is involved in the generation of various life-supporting rhythms. In our research to realize an electronic circuit that mimics the rhythm generating mechanism in the square-wave burster, our circuit experimentally exhibited severe fluctuations in its rhythmic activity. We have found a noise-sensitive region in the phase portrait of the ideal model and have proposed modifications of the model that can reduce this fluctuation. A possible modification to ionic-conductance neuron models (Kohno and Aihara, 2011) was inspired by them. This modification, however, cannot be applied to a group of square-wave bursters, including the Butera–Rinzel–Smith model (0010 and 0050), which is a model of the pre-Bötzinger complex bursting neuron that plays a crucial role in the generation of respiration rhythms, because this modification premises that the slow dynamics originates from an activation gate variable of a hyperpolarizing ionic current. However, in some square-wave bursters, they are controlled by an inactivation gate variable of a depolarizing ionic current. In this study, we proposed a similar modification with a completely different mechanism that can be applied to this group of square-wave bursters. In the presence of noises, the modified Butera–Rinzel–Smith model can generate rhythmic activity that is more stable and similar to biological observations than the original model. The mechanisms underlying this modification are explained with noisy bifurcation diagrams.