Association of MBL gene polymorphisms with major bacterial infection in patients treated with high-dose chemotherapy and autologous PBSCT.

A growing body of evidence indicates that genetic factors are involved in an increased risk of infection. We investigated whether mannose-binding lectin (MBL) gene polymorphisms that cause low levels of MBL are associated with the occurrence of major infections in patients, mainly bearing hematological malignancies, after high-dose chemotherapy (HDT) rescued by autologous peripheral blood stem cell transplantation (auto-PBSCT). A retrospective evaluation of 113 patients treated with HDT and auto-PBSCT revealed that the low-producing genotypes, B/B and B/LXA, were associated with major bacterial infection (P=0.0016, OR 7.9). We next performed a nation-wide large-scale study to assess the allele frequency of the MBL coding mutation in a total of 2623 healthy individuals in Japan. The frequency of allele B was estimated to be approximately 0.2, almost the same in seven different areas of Japan. This common occurrence suggests that MBL deficiency may play an important role in the clinical settings of immunosuppression.

Mutation of DNASE1 in people with systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a highly prevalent human autoimmune diseases that causes progressive glomerulonephritis, arthritis and an erythematoid rash. Mice deficient in deoxyribonuclease I (Dnase1) develop an SLE-like syndrome. Here we describe two patients with a heterozygous nonsense mutation in exon 2 of DNASE1, decreased DNASE1 activity and an extremely high immunoglobulin G titer against nucleosomal antigens. These data are consistent with the hypothesis that a direct connection exists between low activity of DNASE1 and progression of human SLE.

Dominant expression of a novel splice variant of caspase-8 in human peripheral blood lymphocytes.

Caspase-8 is an apical and critical proteolytic enzyme in the cascade of apoptosis. As a result of alternative splicing, the generation of at least 7 isoforms of caspase-8 has been reported. The existence of multiple isoforms that lack the essential domains for apoptosis suggests the possible role of these isoforms on the regulation of apoptosis. Here we report a novel longer isoform of caspase-8 (caspase-8L) that was generated by alternative splicing of intron 8, thereby carrying a 136-bp insertion and frame shift of the transcript. The transcript encoded N-terminal two repeats of death effector domain (DED) of caspase-8, but lacking the C-terminal half of the proteolytic domain. Reverse transcriptase (RT)-polymerase chain reaction (PCR) analysis revealed the dominant expression of caspase-8L transcript compared to the intact form of caspase-8 in human peripheral blood lymphocyte (PBL) and T cells. In patients with systemic lupus erythematosus (SLE), imbalanced expression of caspase-8L transcript was identified. These results suggest the important role of caspase-8L in the modulation of apoptosis.

Mannose binding lectin (MBL) gene mutation is not a risk factor for systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) in Japanese.

Mannose binding lectin (MBL) deficiency may be associated with increased susceptibility to infection and autoimmune disorders, such as systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). In the present study, we performed for the first systematic search for mutations in all the four exons of the MBL gene using polymerase chain reaction (PCR)/single-strand conformation polymorphism (SSCP) analysis. Of 49 healthy Japanese individuals studied, only the previously reported mutation at the codon 54 (substitution from Gly to Asp; G54D) was identified. The allele frequencies of G54D in 105 healthy Japanese individuals, 95 SLE patients and 59 RA patients, were 0.233, 0.226 and 0.178, respectively, which were not significantly different. In addition, two polymorphisms at positions of -550 and -221 in the promoter region were not associated with SLE and RA. It is unlikely that MBL deficiency plays a major role in the pathogenesis of SLE and RA in Japanese.

A novel nonsense mutation at Glu-631 in a Spanish family with complement component 7 deficiency.

Deficiency of the seventh component of complement (C7D) is frequently associated with recurrent neisserial infections. We report in the present study the genetic basis for C7D in a Spanish family. We used exon-specific polymerase chain reaction (PCR)/single-strand conformation polymorphism (SSCP) analysis as a screening step for mutations, followed by direct sequencing of the target exon. The mutation in the proband was a homozygous G-to-T transversion at nucleotide 1957, the first nucleotide of the codon GAG for Glu-631, leading to a stop codon TAG (E631X). Our result provides further evidence that the molecular pathogenesis of C7D is heterogeneous.

A non-sense mutation at Arg95 is predominant in complement 9 deficiency in Japanese.

Deficiency of the ninth component of complement (C9D) is one of the most common genetic abnormalities in Japan, with an incidence of one homozygote in 1000. Although C9D individuals are usually healthy, it has been shown that they have an significantly increased risk of developing meningococcal meningitis. In the present study we report the molecular bases for C9D in 10 unrelated Japanese subjects. As a screening step for mutations, exons 2 to 11 of the C9 gene were analyzed using exon-specific PCR/single-strand conformation polymorphism analysis, which demonstrated aberrantly migrating DNA bands in exon 4 in all the C9D subjects. Subsequent direct sequencing of exon 4 of the C9D subjects revealed that eight of the 10 C9D subjects were homozygous for a C to T transition at nucleotide 343, the first nucleotide of the codon CGA for Arg95, leading to a TGA stop codon (R95X). R95X is a novel mutation different from those recently identified in a Swiss family with C9D. Cases 6 and 7 were heterozygous for the R95X mutation. Family study in case 10 confirmed the genetic nature of the defect. In case 6, the second mutation for C9D of the C9 gene was identified to be the substitution of Cys to Tyr at amino acid residue 507 (C507Y), while the genetic defect(s) in the other allele in case 7 remains unknown. Our results indicate that a novel mutation, R95X, is present in most cases of C9D in Japan.

[Pyloroplasty and motile functions of the stomach and duodenum].

Mongrel dogs were prepared by attaching 2 and 1 silver needle bipolar electrodes on the gastric antrum and bulbus of the duodenum, respectively. Among the obtained active potential from these dogs at chronic stage, investigations were performed mainly on the discharge frequency of spikes, propagation pattern of electrical excitation, and relationship of the stomach and duodenum. In addition, barium was loaded as the stomach contents and extraction dynamics of the loads were observed radiologically. The following was these investigations concerning about the pyloroplasty and motile functions of the stomach and duodenum. Experimental methods 1) Electromyography and extraction dynamics of stomach contents were observed in no treated control dogs. 2) Heineke-Mikulicz type pyloroplasty, Finney type pyloroplasty, or pylorectomy was performed on the above described control dogs. Electromyographical comparison was made among the types of pyloroplasty. 3) Dogs subjected to selective vagolysis (SV), selective proximal vagotomy (SPV), or gastric transection (Tr) were divided into groups with and without pyloroplasty. Electromyography and extraction dynamics of stomach contents were observed in these groups of animals. Results 1) In the control dogs, the mean discharge frequency of spikes in the gastric antrum at fasting time was 4.86 cycle/minute. The spikes propagated to the pyloric side periodically. The discharge frequency of spikes was reduced by the load of stomach contents which took about 120 minutes to be extracted. 2) When the types of pyloroplasty were compared, smaller electromyographical changes were obtained in dogs subjected to Heineke-Mikulicz type pyloroplasty. 3) Pyloroplasty did not affect the discharge frequency of spikes at fasting time, whereas, high incidences of antiperistaltic propagation pattern of electrdcal excitation were observed in dogs subjected to SPV. 4) When stomach contents were loaded, drainage effects of pyloroplasty were suggested from the discharge frequency of spikes, propagation pattern of electrical excitation, and findings in extraction dynamics of the stomach contents. In the dogs subjected to SPV, pylorplasty induced no significant fluctuations in the discharge frequency of spikes. Dogs subjected to Tr showed no improvement in the frequent antiperistaltic propagation pattern of electrical excitement after the pyloroplasty. 5) By pyloroplasty, the frequency of simultaneous generation of spikes in the gastric antrum and duodenum was decreased in the SV and Tr groups, while it was increased in the SPV group.