Concentrated protein body product derived from rice endosperm as an oral tolerogen for allergen-specific immunotherapy--a new mucosal vaccine formulation against Japanese cedar pollen allergy.

The endoplasmic reticulum-derived type-I protein body (PB-I) from rice endosperm cells is an ideal candidate formulation for the oral delivery of bioencapsulated peptides as tolerogens for allergen-specific immunotherapy. In the present study, PBs containing the deconstructed Japanese cedar pollen allergens Cryptomeria japonica 1 (Cry j 1) and Cry j 2 were concentrated by treatment with thermostable α-amylase at 90°C to remove the starch from milled rice powder, which resulted in a 12.5-fold reduction of dry weight compared to the starting material. The modified Cry j 1 and Cry j 2 antigens in this concentrated PB product were more resistant to enzymatic digestion than those in the milled seed powder despite the absence of intact cell wall and starch, and remained stable for at least 10 months at room temperature without detectable loss or degradation. The high resistance of these allergens could be attributed to changes in protein physicochemical properties induced by the high temperature concentration process, as suggested by the decreased solubility of the antigens and seed proteins in PBs in step-wise-extraction experiments. Confocal microscopy showed that the morphology of antigen-containing PB-Is was preserved in the concentrated PB product. The concentrated PB product induced specific immune tolerance against Cry j 1 and Cry j 2 in mice when orally administered, supporting its potential use as a novel oral tolerogen formulation.

[Technical advantages of ultrasound-guided obturator nerve block compared with the nerve stimulating technique].

BACKGROUND: We investigated the onset time of obturator nerve block and time required for the nerve block comparing two groups in a prospective randomized study. METHODS: Obturator nerve block was performed in 37 cases by either nerve stimulation guide (NS group) or nerve stimulation and ultrasound guide (US group) with 10 ml of 1.5% lidocaine adding 1:200000 epinephrine. RESULTS: There was no significant difference between the onset time in the NS group (6.1 min) and that in the US group (5.7 min). The time required for the nerve block (4.2 min vs 1.7 min) was significantly shorter, and the frequency of needle advancement for the optimal position (4.7 vs 1.7) was significantly smaller in the US group than in the NS group (P<0.05). CONCLUSIONS: Ultrasound imaging facilitated the identification of obturator nerves between adductor muscles. Ultrasound-guided obturator nerve block is a safe, quick and useful technique.

[Anesthetic management by psoas compartment block for adult inguinal herniorrhaphy].

Spinal anesthesia has been often used for inguinal herniorrhaphy. With the advancement of surgical technique, tension free inguinal herniorrhaphy is now performed under local anesthesia. Local anesthesia, however, does not always offer sufficient anesthesia. We performed psoas compartment block with 1% lidocaine in a 78-year-old man undergoing tension free inguinal herniorrhaphy. During the operation, the patient felt pain and needed additional local anesthesia once. But the pain was not severe, and he did not feel stressed. Advantages of psoas compartment block for inguinal herniorrhaphy are its easiness to perform, little burden of anesthesia and high efficacy. Demerits are slow onset of block and necessity for prolonged patient monitoring. Further investigations on dose and patient position are required for psoas compartment block to become useful for inguinal herniorrhaphy.

cDNA cloning, gene structure, and expression of Broad-Complex (BR-C) genes in the silkworm, Bombyx mori.

To clarify the molecular mechanisms of metamorphosis, we analyzed the Broad-Complex (BR-C) gene in the silkworm, Bombyx mori. We cloned cDNAs for the full coding regions of the Z1, Z2, and Z4 isoforms of BR-C. The Z3 zinc finger sequence was found in the 3'UTR of the Z2 isoform. The predicted amino acid sequence showed high homology with Drosophila and Manduca BR-C proteins. Five bacterial artificial chromosome (BAC) clones were screened from a Bombyx BAC library. Restriction enzyme cleavage maps of 170 kb regions were constructed, and a total of 25.8 kb were sequenced. The BAC analysis showed that the 5'UTR of the BR-C gene consists of the first two exons, while the coding region contains a core region domain with five exons and four zinc finger exons in the order Z1, Z4, Z2, and Z3. Expression analysis revealed 9.5, 6.5, and 5.5 kb BR-C transcripts. These increased during the spinning ecdysone peak on day 6 of the fifth instar when pupal commitment occurs in the Bombyx epidermis. In addition, a small amount of BR-C mRNA was detected in the epidermis before this peak. BR-C mRNA was also expressed in the fat body from day 1 in the fourth instar to day 7 in the fifth instar.