Polymorphisms in the TNFA promoter region is not associated with palmoplantar pustulosis.

Polymorphisms of the 5'-flanking promoter/enhancer region of the TNAFA gene were determined in 80 Japanese patients with pulmoplantar pustulosis (PPP). The 5'-flanking region of the TNFA gene from -1107 to 66 was amplified by polymerase chain reaction (PCR) method. Nucleotide sequencing data from the PCR products revealed that 5 single nucleotide polymorphisms at position 1031, -863, -857, -307 and -237. None of the nucleotide substitutions were significantly increased in PPP patients when compared with those in controls. To clarify the linkage among the neighboring genetic marker, we analyzed the association between the polymorphisms in the TNFA promoter region and the NcoI polymorphism in the first intron of the TNFB gene as well as HLA-DR9. The genotype at 1031C is strongly associated with TNFB1 and negatively associated with TNFB2 which is reported to be associated with PPP. These data indicate that TNFA gene centromeric to TNFB is not associated with PPP and the susceptible gene of PPP is located between TNFB and HLA-B.

Polymorphism in the tumor necrosis factor B gene is associated with Palmoplantar pustulosis.

We investigated the allele and genotype distribution of a polymorphism of the tumor necrosis factor (TNF) B gene and the frequency of HLA-DR9 in 49 patients with Palmoplantar pustulosis (PPP) and 51 healthy controls. We found that the frequency of TNFB2 in the PPP patients was significantly higher than that in the controls. Furthermore, the DR9-TNFB2 haplotype was significantly more frequent in the PPP patients (P=0.0045). These results suggest that TNFB2 may confer susceptibility to PPP.

Tolerogenic capacity of poly(ethylene glycol) (PEG)--modified ovalbumins in relation to their immunoreactivity towards anti-ovalbumin antibody.

Ovalbumin (OVA) was chemically modified with poly(ethylene glycol) (PEG) derivatives: activated PEG2, 2,4-bis[O-methoxypoly(ethylene glycol)]-6-chloro-s-triazine and activated PM13, copolymer of poly(oxyethylene) allyl methyl diether and maleic anhydride. Pretreatment of BALB/c mice with PEG2- or PM13-OVA suppressed the production of both IgG- and IgE-class anti-OVA antibodies induced by subsequent immunizations with unmodified OVA. PEG2-OVA had higher potency to suppress OVA-specific immune response than PM13-OVA. Although extensive modification (62%) of amino groups in the OVA molecule with activated PEG2 was required to diminish most of its immunoreactivity towards anti-OVA antibodies, only a low degree of modification (27%) was sufficient to induce tolerogenicity to OVA. Thus, the loss of immunoreactivity of PEG2-OVA was not prerequisite for its tolerogenic capacity. Moreover, the use of completely denatured PEG2-OVA immunogen lead to the loss of its ability to induce immune tolerance to native OVA. These observations are discussed in relation to the regulatory mechanism of immune response.

Intracellular vesicular stomatitis virus nucleocapsids and virions visualized by surface spreading.

Spreading of cells on a solution surface could visualize vesicular stomatitis virus nucleocapsids and virions in infected cells easily and clearly without the need for any purification. Characteristic structures observed by the spreading of the infected cells are described and discussed.

Some applications of the surface-spreading technique to virological studies.

By application of the surface-spreading technique, virus particles in infected cells and viremia serum, and precipitates in agar plates of the double immunodiffusion technique of Ouchterlony could easily and clearly be visualized without any purification process.

Process of cap formation of messenger RNA by vaccinia virus particles carrying an organized enzymes system.

Vaccinia virus mRNAs carry the cap structure m7G5'pppAm- or m7G5'pppGm- at the 5'-terminus, which is synthesized by a series of RNA polymerase and capping enzymes contained in the virus particle. The process of the cap formation at the 5'-terminus of mRNA was studied using an in vitro system under similar conditions to those of vaccinia virus multiplication in its host cell. After adding a methyl-group donor, methyl-3H-S-adenosylmethionine, the oligonucleotides, which were the de novo synthesized 5'-terminal part of mRNA, were isolated from the RNA-synthesizing virion at appropriate time intervals, and were analysed. The 5'-5' confronting nucleotides with 2'-O-methylation, G5'pppAm and G5'pppGm, were found with the completed cap structure, m7G5'pppAm and m7G5'pppGm. The confronting nucleotides with only 7-methyl guanine as a methylated component, m7G5'pppA and m7G5'pppG, were not detected at any incubation time, and it was concluded that methylation at the 2'-position of the 5'-terminal purine nucleoside of mRNA precedes methylation of the 7-position of the blocking guanosine. This result is different from that obtained using the enzymes isolated from vaccinia virus (Moss et al., 1976) and also from the results obtained using other kinds of virus particles, which carry RNA polymerase and capping enzymes. These differences may be due to the specific organization of a series of capping enzymes and RNA polymerase in each virus particle.

Methylation of 5'-5' confronting dinucleotides by vaccinia associated enzyme system.

Methylation of a series of the 5'-5' confronting dinucleotides containing various number of phosphates was examined by the use of virion enzyme system of vaccinia virus. Methylation was absolutely dependent on the number of interposed phosphate groups between 5'-5' confronting dinucleotides, showing maximum efficiency in the 3 phosphate compound corresponding to the blocked 5'-terminus (cap) of an eukaryotic mRNA. Methylation, if any, occurred only at the 7-position of guanine residue, but not at the 2' position of ribose moiety.

[Clinical studies on PC-904 in biliary tract diseases (author's transl)].

The clinical effects of PC-904, a new semisynthetic penicillin, were studied in patients with biliary tract diseases, and the results were as follows: 1) PC-904 showed an average peak serum level of 40.7 +/- 11.6 microgram/ml 2 hours after an intravenous drip infusion of 1 g of the agent. The biliary level showed a peak value of 126.5 +/- 85.4 microgram/ml 2 hours to 3 hours after the infusion. 2) Isolated organisms from bile before the treatment were E. coli, Klebsiella, Proteus, Pseudomonas, Enterococcus and Bacteroides. MIC of PC-904 on 18 strains of isolated organisms was almost 6.25 microgram/ml or less. All isolated organisms except one strain of Klebsiella oxytoca disappeared after the treatment. 3) Six patients with cholelithiasis were medicated with PC-904 to prevent post-operative infections. The clinical effects were good in 4, poor in 1 and unknown in 1 case. 4) As to side effects no adverse reactions and allergic reactions were noted. Also no significant abnormalities of laboratory findings were observed.

Pteridines produced by Methylococcus capsulatus. Isolation and identification of a neopterin 2':3'-phosphate.

Three pteridines have been isolated from the methane- or methanol-oxidizing bacterium Methylococcus capsulatus. Two of these are known compounds, 2-amino-6-carboxy-4-hydroxypteridine and 2-amino-4-hydroxy-6-methylpteridine. The third is shown by degradative and synthetic experiments to be l-threo-neopterin 2':3'-phosphate. Labelling experiments show that both the pteridine moiety and phosphate residue are derived from a single GTP molecule. The possible metabolic significance of these compounds in methanol oxidation is discussed.