Purification and immunochemical characterization of alpha-fetoprotein from rat fetal serum and liver

Two alpha1-globulin bands of fetal serum with relative mobilities against bromophenol blue of 0.55 and 0.58 on 7% polyacrylamide gel electrophoresis reacted with a monospecific rabbit antiserum to alpha-fetoprotein (AFP). The former globulin band was clearly detected in the fetal liver supernatant. AFP was immunochemically purified from both the fetal serum and liver, and their electrophoretic and immunochemical properties were compared. Liver AFP purified by immunoadsorbent column yielded electrophoretic mobilities and relative amounts of the two electrophoretically distinct components identical with the purified serum AFP. The immunological reactivity of the two components of the purified preparations from serum and liver against the monospecific anti-AFP serum was also indistinguishable. After the removal of the sialic acid residues from purified serum and liver AFP by treatment with neuraminidase for 6 to 12 hr, disc electrophoretic patterns on 5% polyacrylamide gel and immunoelectrophoretic patterns of the treated AFP were found to be closely similar in both preparations. It may be possible to conclude that serum and liver AFP are structurally indistinguishable and probably identical.</p

Non-LTE Line-Formation and Abundances of Sulfur and Zinc in F, G, and K Stars

Extensive statistical-equilibrium calculations on neutral sulfur and zinc were carried out, in order to investigate how the non-LTE effect plays a role in the determination of S and Zn abundances in F, G, and K stars. Having checked on the spectra of representative F-type stars (Polaris, Procyon, and alpha Per) and the Sun that our non-LTE corrections yield a reasonable consistency between the abundances derived from different lines, we tried an extensive non-LTE reanalysis of published equivalent-width data of S I and Zn I lines for metal-poor halo/disk stars. According to our calculations, S I 9212/9228/9237 lines suffer significant negative non-LTE corrections amounting to <~ 0.2--0.3 dex, while LTE is practically valid for S I 8683/8694 lines. Embarrassingly, as far as the very metal-poor regime is concerned, a marked discordance is observed between the [S/Fe] values from these two abundance indicators, in the sense that the former attains a nearly flat plateau (or even a slight downward bending) while the latter shows an ever-increasing trend with a further lowering of metallicity. The reason for this discrepancy is yet to be clarified. Regarding Zn, we almost confirmed the characteristic tendencies of [Zn/Fe] reported from recent LTE studies (i.e., an evident/slight increase of [Zn/Fe] with a decrease of [Fe/H] for very metal-poor/disk stars), since the non-LTE corrections for the Zn I 4722/4810 and 6362 lines (tending to be positive and gradually increasing towards lower [Fe/H]) are quantitatively of less significance (<~ 0.1 dex).Comment: 33 pages, 7 figures, PASJ, Vol. 57, No. 5 (2005) in pres

Dynamic Deformation Characteristics of Rockfill Materials from Laboratory Test, In-Situ Test and Earthquake Motion Analysis

On dynamic analyses of rockfill dams, dynamic deformation characteristics of rockfill materials such as shear moduli and damping ratios must be known to make analysis more accurate. In this study, large-scale cyclic triaxial tests were carried out using rockfill materials of actual dams, and the results were compared with the dynamic deformation characteristics obtained by in-situ geophysical explorations and response analyses of earthquake motions observed at dams. Furthermore, the radiation damping ratio was estimated from response analyses and laboratory tests, and then the frequency and strain dependency characteristics of the radiation damping were evaluated

Activation of cAMP-dependent Protein Kinase in Epidermis by the Compounds which Increase Epidermal cAMP

Pig epidermal slices were incubated with various compounds which increased epidermal cAMP (adenosine 3',5'-monophosphate), and the change in cAMP-dependent protein kinase activity ratio was studied by the method of Cherrington et al (J Biol Chem 251:5209–5218, 1976) with modification.Epinephrine (5 × 10−5 m), histamine (10−4 m) and adenosine (10−3 m), potent agonists of epidermal adenyl cyclase, fully activated the protein kinase (PK) during an incubation of 30 to 45 seconds, that was much shorter than that required for maximal cAMP accumulation under the same conditions (5 min). With such a brief stimulus, the epidermal cAMP-PK system did not become refractory and responded to repeated stimuli. Prostaglandin E2 (PGE2) and isobuthylmethylxanthine (IBMX) and ethanol only partially activated the enzyme. Prostaglandin F2α. (PGF2α) and theophylline which were much less effective in increasing epidermal cAMP, activated the enzyme to the same extent as PGE2 and IBMX respectively.These results suggest that protein kinase activation takes place in response to a cAMP increase in small locus of the cell. Such an increase in cAMP can be very small or even not measurable when measured as total cAMP in the tissue homogenate. Also, increases above this level may not be physiologic.It is concluded that measurement of cAMP-dependent protein kinase activity ratio is a more direct and more sensitive way to study the effect of compounds which act through cAMP mediated mechanism

Effects of mesophyll water potential on photosynthesis in Cyperaceae plants: with special reference to phylogeny of tribes and decarboxylation sub-types

We examined the photosynthetic rates under water stress conditions in 43 Japanese Cyperaceae species using the same method used for Gramineae plants. Compared with Gramineae, the difference between C4 and C3 species was more distinct in Cyperaceae. Moreover, C4 Cyperaceae species were very susceptible to water stress like Panicoideae C4 species. These species belong to the NADP-ME subtype. It appears that the sensitivity of photosynthesis to water stress would be different depending on the decarboxylation sub-types

Phosphorylation of Pig Epidermal Soluble Protein by Endogenous cAMP-Dependent Protein Kinase

The distribution of adenosine 3',5'-monophosphate (cAMP)-dependent protein kinase and its substrate proteins was analyzed using soluble and particulate fractions of pig epidermal homogenates. When histone was used as a substrate for this enzyme reaction, protein kinase activity was distributed almost equally between the soluble and particulate fractions. However, the effect of exogenously added cAMP was confined almost exclusively to the soluble enzyme. Endogenous protein phosphorylation in the absence of exogenous histone was higher in the particulate fraction than in the soluble fraction, but the stimulating effect of cAMP was observed only in the soluble fraction. These results indicate that cAMP-dependent protein kinase is predominantly localized in the soluble fraction and phosphorylates soluble epidermal proteins. The particulate fraction contains protein kinase which is cAMP-independent and phosphorylates particulate-bound proteins as well as histone. Based on these observations, the soluble fraction was incubated with [γ-32P]-ATP in the presence or absence of cAMP, and phosphorylated protein was analyzed by SDS disc- or slab-gel electrophoresis followed by autoradiography. Among many proteins whose phosphorylation was slightly increased by cAMP, a protein with Mr ∼45,000 was found which was markedly phosphorylated in the presence of cAMP. Although this protein corresponds to one of the richest proteins in the epidermal soluble fraction, an important physiologic role for this phosphorylation has not been clarified

Up-Regulation of the Brain and Purkinje-Cell Forms of Dystrophin Transcripts, in Becker Muscular Dystrophy

Farrando Sicilia, Jordi; Llauradó Grau, Josep M. ; Fuente Fuente, Carlos; Montes, Antoni

Cyclic AMP-Dependent Protein Kinase Isozymes of Pig Skin and Human Skin from Normal and Psoriatic Subjects

Cyclic AMP-dependent protein kinase isozymes of pig and human skin (epidermis) were separated by DEAE- cellulose column chromatography after micromodification for small biopsy samples. Clear-cut separations of type I and type II isozymes, winch were of about equal amounts, could be obtained only when the ischemia effect was avoided by in vivo freezing of skin and homogenization for less than 10 s. Intradermal injections of epinephrine caused dose-dependent activation of type I isozyme, but not of type 11. Injections of other skin adenylate cyclase stimulators such as histamine, adenosine, and prostaglandin E2 elevated the local cyclic AMP levels to not more than 5 pmol/mg protein and also stimulated only the type I isozyme. Incubation of keratome-sliced pig skin under various conditions caused both activation by dissociation and inactivation by dissociation of the subunits, which appeared to be dependent on the cyclic AMP content. Epinephrine added to the incubation medium led to complete activation of both type I and type II isozymes (the intraepidermal cyclic AMP contents ranged from 20–50 pmol/mg protein). The isozymes of normal skin and involved skin of psoriatics showed identical peaks of type I and type II Isozymes of equal amounts. The data indicate that protein kinase in the involved skin is not in an activated (by cyclic AMP) state