Reversed-phase high-performance thin-layer chromatography of hematoporphyrin and its nickel,copper and zinc complexes

Summary The thin-layer chromatographic mobility of hematoporphyrin IX (Hp) and its metal complexes on octyl (C₈)- or octadecyl (C₁₈)-bonded silica gel plate is described. The mobility order for those compounds is regular on both kinds of plate with various compositions of methanol-phosphate buffer mixtures as the developing solvents; thus, Cu-complex < Ni-complex < Hp (free acid) < Zn-complex. The combination of a C₁₈-plate with an 85∶15 (vol/vol) mixture of methanol and phosphate buffer (pH 3) is recommended for the successful separation of these four compounds.     

Amidase and peptidase activities of polyclonal immunoglobulin G present in the sera of patients with rheumatoid arthritis

Polyclonal Immunoglobulin (Ig) G from patients with rheumatoid arthritis (RA) and healthy subjects hydrolyzed carbobenzoxy−Val−Gly−Arg p-nitroanilide and D−Pro−Phe−Arg p-nitroanilide. RA IgG exhibited higher activity against the former substrate, but not the latter. On the other hand, RA IgG showed reduced activity against D−Pro−Phe−Arg methylcoumarinamide, when compared with those of the healthy controls. These results suggest that RA IgGs differ from normal IgGs in the substrate specificity of amidase activity. Preliminary studies have shown that two out of three RA IgG samples cleaved a pentapeptide—Gln−Arg−Arg−Arg−Ala−Ala— which is assumed to be associated with the risk of developing RA (Gregersen, P. K. et al. (1987), Arthritis Rheum. 30, 1205–1213). By contrast, virtually no cleavage of the same peptide was observed with IgG from healthy controls. A peptide analog, Gln−Arg−Arg−Trp−Ala, was not cleaved at all by any IgGs examined either from RA patients or healthy controls.     

Steroid Cyclophanes as Artificial Cell-surface Receptors. Molecular Recognition and its Consequence in Signal Transduction Behavior

Steroid cyclophanes, bearing four bile acid moieties covalently placed on a tetraazaparacyclophane skeleton, were designed and synthesized as artificial cell-surface receptors. Guest-binding behavior of the steroid cyclophanes embedded in a bilayer membrane formed with a synthetic peptide lipid was clarified by means of fluorescence and circular dichroism spectroscopy. We found that the steroid cyclophane effectively bound aromatic guests in both bilayer membranes and aqueous solution. In addition, copper(II) ions acted as a guest species for the steroid cyclophane and a competitive inhibitor toward a NADH-dependent lactate dehydrogenase (LDH). On these grounds, we constituted a supramolecular assembly as an artificial signaling system in combination with the steroid cyclophane, a cationic peptide lipid, and LDH. As a consequence, the steroid cyclophane acted as an effective artificial cell-surface receptor being capable of transmitting an external signal to the enzyme in collaboration with copper(II) ions as a signal transmitter.     

Studies on cerebral protective agents. I. Novel 4-arylpyrimidine derivatives with anti-anoxic and anti-lipid peroxidation activities.

Novel 4-arylpyrimidine derivatives were synthesized by the oxidation of 4-aryl-1,4-dihydropyrimidines, and their effects on anti-anoxic (AA) activity in mice and anti-lipid peroxidation (ALP) activity in rat brain mitochondria were investigated. Among these compounds, ethyl 6-methyl-2-phenyl-4-(4-pyridyl)-5-pyrimidinecarboxylate (4b) has AA activity (10 mg/kg, i.p.) and ethyl 6-methyl-4-(3-nitrophenyl)-2-phenyl-5-pyrimidinecarboxylate (4f) has ALP activity (73% inhibition at 10(-5) g/ml). The latter compound (100 mg/kg, i.p.) was also effective on arachidonate-induced cerebral edema in rats with comparable potency to that of vitamin E.     

Characterization of the supermolecular structure of cellulose in wood pulp fibres

The hierarchic organization of cellulose fibrils (microfibrils) was investigated in holocellulose, sulphite pulp and kraft pulp using TEM, XRD, ED and FTIR. There were remarkable differences in both the fibril structure and fibril aggregation between the samples. TEM observations revealed more intimately associated fibrils in the kraft pulp compared to the sulphite pulp and the holocellulose, results in agreement with previous CP/MAS ¹³C-NMR data [Hult E.-L. et al. (2002) Holzforschung 56: 231–234]. Furthermore, the cellulose crystallinity was higher in the kraft pulp sample. With respect to the cellulose I and I allomorphs, these samples were controversial when different analytical techniques were applied. Due to the small fibril size and the low degree of order of cellulose in these samples, the concept of crystalline triclinic and monoclinic components as determined by diffraction analysis may not be adequate. Instead the fibril can be regarded to have different degrees of lateral order (including paracrystalline ordering) that can be reoriented to I type conformation and packing upon pulping.     

Cp/mas 13c nmr and Electron Diffraction Study of Bacterial Cellulose Structure Affected by Cell Wall Polysaccharides

Acetobacter xylinum was cultured in Schramm–Hestrin medium containing pectin (pectin medium), xylan (xylan medium), or glucomannan (mannan medium). X-ray diffractometry revealed that xylan and glucomannan affected the size of the cellulose crystals and their d-spacing values. Solid-state cross polarization magic angle spinning carbon-13 nuclear magnetic resonance spectroscopy indicated that the ratio of cellulose I was reduced by the addition of polysaccharides. These effects were more remarkable on the cellulose in the mannan medium than that in the xylan medium, and were scarcely observed in the pectin medium. Electron diffraction analysis revealed that these effects on hemicelluloses along cellulose microfibrils are continuous in the mannan medium and discontinuous in the xylan medium. These findings suggest that the uronic acid in the polysaccharides prevents interactions with cellulose leading to alterations of the structure of the cellulose crystal.     

Determination of phosphorus in biological material using electrothermal atomisation atomic absorption spectrometry with a molybdenum tube atomiser

Phosphorus has been determined by electrothermal atomisation atomic absorption spectrometry with a molybdenum tube atomiser. The effects of interferents on the phosphorus signal have been investigated. Chemical interferences were eliminated by high-temperature pyrolysis. The detection limit for the molybdenum tube system was 2.3 ng (corresponding to 2.3 micrograms ml-1 when 1 microliter was injected into the atomiser) and the characteristic mass was 1.9 ng of phosphorus. A calibration graph method combined with high-temperature pyrolysis has been evaluated through the determination of phosphorus in biological materials. The sensitivity, accuracy and precision of the method were superior to or nearly equal to those of graphite furnace atomic absorption spectrometry. The advantages of the high-temperature pyrolysis method are its simplicity and low cost.     

DNA cleavage reaction of antitumour antibiotic, kapurimycin A3, with deoxytetranucleotide d(CGCG)2

Kapurimycin A3 (kap A3, 1 ), an antitumour antibiotic, alkylates N7 of guanine₂ (G₂) and G₄ of d(C₁G₂C₃G₄)₂ to produce their covalent adducts 2 (64 %) and 3 (7.0 %), respectively. Heating at 90 °C for 5 min degraded both adducts to kap A3 - G adduct (5) with the concurrent release of their respective abasic-site containing oligomers 4 and 6.     

Fluorescence immunological determination of immunoglobulin G in human serum by high-performance liquid gel-permeation chromatography

Summary A high-performance liquid gel-permeation chromatographic method is described for the determination of human serum immunoglobulin G (IgG) by separating the fluorescent immuno complex from the free fluorescence-labeled antibody. Fluorescence-labeled antibody used in this study was fluorescein isothiocyanate (FITC)-labeled Fab fragment goat anti-human IgG (anti-IgG Fab). Immuno complexes and antibody of different molecular sizes can be separated. FITC-labeled anti-IgG Fab was added to the serum and the mixture is passed through the column. An immuno complex separates as well-delineated peak in the column void volume, and was measured by the fluorescence of the column eluate (Ex=490nm, Em=520nm). The total analysis time for a serum sample was approximately 15min. The minimum detection limit was 25 mg/dl. The relative standard deviation was below 2% (peak area). The results of the HPL-GPC analysis correlate well with those obtained by laser nephelometric assay (r=0.992).