Human aldolase isozyme gene: the structure of multispecies aldolase B mRNAs.

A complete nucleotide sequence of human aldolase B mRNA was determined with a recombinant cDNA (pHABL120-3). The cDNA insert was composed of 1,652 bases excluding poly(A) tail and the sequence was consistent with the previous results reported by others. However, S1 nuclease mapping and subsequent genomic analysis allowed us to know that the clone possesses two more sites corresponding to 5'-termini in the 5'-noncoding region and another site of polyadenylation in the 3'-noncoding region. In fact, the major aldolase B mRNA species occupying 90% of the total mRNAs initiated at the predominant position corresponding to the position around -82 of the 5'-noncoding sequence in pHABL120-3 and terminated at the distal polyadenylation site. Second species accounting for 9% of the mRNAs initiated at the same site and terminated at the proximal polyadenylation site. The remainings have a longer 5'-noncoding sequence which starts from further upstream region of the major one and pHABL120-3 corresponds to one of these largest clones.     

Neutral glycosphingolipids of ova of the fresh-water bivalve, Hyriopsis schlegelii

The neutral glycosphingolipids of ova of the fresh-water bivalve, Hyriopsis schlegelii were characterized. The most abundant glycolipid was ceramide monosaccharide, followed by ceramide trisaccharide, ceramide tetrasaccharide, and ceramide disaccharide. More complex neutral glycolipids accounted for almost one-third of the total. The total amount of these glycolipids was 0.59 mg/g of dry weight of the ova preparation, a yield which was one-seventh of that of spermatozoa neutral glycolipids. Structural analyses were performed by enzymatic hydrolysis of the glycolipids with exoglycosidases, permethylation experiments, and also immuno-chemical assays. The proposed structures are as follows: ceramide monosaccharides, Gal-Cer and Glc-Cer; ceramide disacharides, Gal(beta 1-4)Gal-Cer, Gal(beta 1-4)Glc-Cer, and Man(beta 1-4)Glc-Cer; ceramide trisaccharide, Man(alpha 1-3)Man(beta 1-4)Glc-Cer; ceramide tetrasaccharides, Man(alpha 1-3)[Xyl(beta 1-2)]Man(beta 1-4)Glc-Cer, GlcNAc(beta 1-2)Man(alpha 1-3)Man(beta 1-4)Glc-Cer, Man(alpha 1-3)[Gal(beta 1-2)]Man(beta 1-4)Glc-Cer, and Man(alpha 1-2?)Man(alpha 1-3)Man(beta 1-4)Glc-Cer. The latter two ceramide tetrasaccharides were new types of glycosphingolipids. The spectrum of ova glycolipids appeared to be more complicated than that of the spermatozoa glycolipids. The ova glycolipids characterized here, with the exception of ceramide tetrasaccharides, contained considerable amounts of 2-hydroxy fatty acids, which were not observed in the spermatozoa glycolipids. The major sphingosine base was C18-sphingenine in all the ova glycolipids as well as in the spermatozoa glycolipids. However, the content of anteiso type of sphingosine base was 2- to 3-fold higher in the ova than in the spermatozoa.     

Alterations in Fc receptor function of macrophages from streptozotocin-induced diabetic rats

The presence of elevated levels of circulating immune complexes in diabetic humans and animals suggests impaired phagocyte function. To evaluate FcR-mediated phagocytosis, resident peritoneal macrophages were harvested from control, streptozotocin-induced diabetic, and insulin-treated diabetic rats. FcR number and avidity were determined from Scatchard analysis of binding of 125I-labeled aggregated rat IgG (ARG) to macrophages. The total and fractional catabolic capacity were determined by quantitating the digestion of ARG as a percent of the total ARG added and as a percent of ARG bound. Insulin-deficient diabetic rats had an increase in the number of FcR per cell (26.8 +/- 3.5 X 10(4)) as compared with control animals (13.1 +/- 1.2 X 10(4)) (p less than 0.01). In contrast, insulin-treated diabetic animals had a reduction in the number of FcR per cell (9.8 +/- 1.4 X 10(4)) (p less than 0.01). FcR of macrophages from insulin-deficient diabetic rats had a lower avidity (Kd = 6.9 +/- 1.8 X 10(-10)M) when compared with control (3.7 +/- 0.6 X 10(-10)M) and insulin-treated diabetic rats (3.6 +/- 0.9 X 10(-10)M) (p less than 0.01). Total catabolism of ARG by macrophages from both insulin-deficient and insulin-treated diabetic rats was reduced (31.0% +/- 3.4 and 17.5% +/- 3, respectively) when compared with controls (49.6% +/- 5.2) (p less than 0.01). Fractional catabolism by macrophages from insulin-deficient diabetic rats was significantly reduced (21% +/- 1.9 and 4.6% +/- 0.9/10(4) FcR) when compared with results from control rats (26% +/- 1.3 and 6.7% +/- 0.7/10(4) FcR) (p less than 0.01), whereas the results from insulin-treated diabetic rats (32% +/- 2.4 and 10.8% +/- 1.0/10(4) FcR) (p less than 0.01) were greater than those from controls. These studies demonstrate that FcR-mediated phagocytosis of soluble, "model" immune complexes is impaired in macrophages from both insulin-deficient and insulin-treated diabetic rats; however, different mechanisms account for this impairment in phagocytosis. Despite an increase in FcR number of macrophages from insulin-deficient diabetic rats, the depression of post-receptor-mediated catabolism results in a net depression in phagocytic activity. In contrast, macrophages from insulin-treated diabetic rats display augmented post-receptor-mediated catabolism; however, this does not overcome the low initial binding of ARG to the cell that results from the depression of FcR number.     

Effect of high carbohydrate diet on serum gamma-glutamyl transpeptidase fraction pattern in Japanese young men

Changes in the activity of serum gamma-glutamyl transpeptidase (gamma-GTP) and the percentage of the gamma-GTP fraction in healthy young men given a high carbohydrate diet (480-636 g/day, 80% of the total energy) for 21 days were examined. Serum total gamma-GTP activity showed no significant change in four healthy young volunteers who received high carbohydrate diet for 21 days. However, the percentage of the gamma-GTP (1) fraction increased significantly (P less than 0.01) from the basal level of 55.6 +/- 4.0% to 67.6 +/- 0.9% on day 10, and then decreased to 58.4 +/- 1.4% on day 21. When the experimental diet was replaced by usual diet, the percentage of the gamma-GTP (1) fraction returned to the same level as before the experiment. It is concluded from the results that the nutrient intake affects the percentage of gamma-GTP (1), but not the total serum gamma-GTP activity.     

Conversion of 4-aminobutyraldehyde to γ-aminobutyric acid in striatum treated with semicarbazide and kainic acid

4-Aminobutyraldehyde (ABAL) has been shown to cross the blood-brain barrier and to be converted rapidly to -aminobutyric acid (GABA) in various regions of the brain. In this paper, the formation of GABA from ABAL was studied with striatum that had suffered a lesion to GABA synthesis via glutamic acid decarboxylase (GAD). The GABA formation from ABAL was invariably observed in striatum in which GAD was severely inhibited by semicarbazide or kainic acid. Thus, this is another pathway for GABA formation.     

Analysis of powder and single crystal electron paramagnetic resonance spectra of deoxy myoglobins containing cobalt (II) proto-and mesoporphyrins IX at various microwave frequencies

Artificial myoglobins (Mbs) substituted for protoheme with Co(II) proto-and mesoporphyrins IX (proto-and meso-CoMbs, respectively) were prepared. The principal values and eigenvectors of g tensors and the hyperfine coupling tensors of the paramagnetic Co(II) centers of their deoxy forms have been determined by single crystal EPR spectroscopy at 77 K in order to elucidate orientation and electronic structure of the prosthetic group in myoglobin. The orientation of the porphyrin plane of deoxy meso-CoMb were found to be identical to that of deoxy proto-CoMb. However, the in-plane hyperfine coupling constants of deoxy meso-CoMb were more anisotropic and larger than those of deoxy proto-CoMb, suggesting an increase in the electron spin density on the Co(II) ion upon the exchange of protoporphyrin IX with mesoprophyrin IX. Powder EPR spectra of these CoMbs, which were measured at S- and L-band microwave frequencies, exhibited well resolved 59Co hyperfine splittings and can be clearly interpreted by the use of the EPR parameters obtained from single crystal EPR measurements.     

Fresh-water planarias and a marine planaria are relatively dissimilar in the 5S rRNA sequences.

The nucleotide sequences from fresh-water Dugesia japonica and marine Planocera reticulata have been determined. The similarity between these two species is only 69%. The Planocera sequence reveals nearly 80% similarity (72-81%) to the sequences of multicellular animals, while the Dugesia sequences are considerably different from them (66-73%).     

Some properties of salivary amylases of Adelphocoris suturalis (Miridae), Dolycoris baccarum (Pentatomidae), and several other heteropteran species

Some properties of salivary amylases of the adults of Adelphocoris suturalis Jakovlev and Dolycoris baccarum L. were studied in vitro and compared with those of several other heteropteran species. The reducing sugar produced by the action of the salivary amylase of D. baccarum increased in proportion to substrate concentration while the concentration was relatively low (below 0.67% in its final concentration). Its increase stopped at the concentrations from 0.67 to 2.0%, and then it increased again constantly but slowly. The optimum temperature for the action of the enzyme was found to be about 50° (in A. suturalis) and about 40° (in D. baccarum), and the optimal pH was 3.5–4.0 in A. suturalis and 6.0 in D. baccarum. The salivary amylase activity of D. baccarum was scarcely affected by NaCl, KNO₃, and other compounds, while the salivary amylase of A. suturalis was strongly activated by NaCl and moderately by KNO₃. It seems highly probable that taxonomically related heteropterous insects might well be grouped with respect to the degree of activation of the salivary amylase by Cl- or NO₃-.

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Zusammenfassung Einige Eigenschaften der Speichelamylase von Adelphocoris suturalis und Dolycoris baccarum wurden im Reagenzglas studiert und mit denen anderer Heteropterenarten verglichen. Die durch die Wirkung der Speichelamylase von D. baccarum produzierten Stärkehydrolysate vermehrten sich im Verhältnis zur Substratkonzentration bei einer verhältnismäßig niedrigen Konzentration (weniger als 0,67% in der Schlußkonzentration). Diese Vermehrung wurde bei einer Konzentration von etwa 0,67 bis 2,0 unterbrochen, um sich dann wieder stetig, aber langsamer fortzusetzen. Die optimale Temperatur für die Wirkung des Fermentes lag für A. suturalis bei etwa 50° und für D. baccarum bei etwa 40°. Die optimale Wasserstoff-Ionenkonzentration betrug für A. suturalis pH 3,5–4,0 und für D. baccarum pH 6,0. Die Aktivität der Speichelamylase von D. baccarum wurde durch NaCl, KNO₃ und andere Verbindungen kaum beeinflußt, während die Speichelamylase von A. suturalis durch NaCl stark und durch KNO₃ schwach aktiviert wurde. Es wird vermutet, daß, hinsichtlich des Aktivierungsgrades der Speichelamylase durch Cl- und NO₃-, systematisch nahe verwandte Heteropterenarten der gleichen Gruppe angehören.

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Contribution No. 28 from the Laboratory of Entomology, Obihiro Zootechnical University.