Aquaporin and aquaglyceroporin in silkworms, differently expressed in the hindgut and midgut of Bombyx mori.

Two cDNAs similar to aquaporins (AQPs) from other insect species were identified and characterized from the silkworm larva, Bombyx mori. The first cDNA (AQP-Bom1) cloned from the anterior silk gland encodes a 25 900 Da protein similar to insect AQPs isolated from several liquid-feeding insects. The second cDNA (AQP-Bom2) cloned from the posterior midgut encodes a 27 694 Da protein. Northern blot analysis has revealed that the AQP-Bom1 mRNA (2.3 kb) is expressed predominantly in the hindgut (colon and rectum), and moderately or minimally in the silk gland, midgut and Malpighian tubules, while the AQP-Bom2 mRNA (1.3 kb) is mainly expressed in the posterior midgut and Malpighian tubules. Functional analysis in Xenopus oocytes microinjected with the cRNA of these AQPs revealed that the AQP-Bom1 mRNA encodes a water-specific aquaporin, likely involved in the water retrieval function of the hindgut, while the AQP-Bom2 mRNA encodes an aquaglyceroporin, increasing glycerol and urea uptake.

CYP19A1 genetic polymorphisms may be associated with obesity-related phenotypes in Chinese women.

OBJECTIVE: To examine the relationship between genetic polymorphisms of the CYP19A1 gene and obesity-related phenotypes, body mass index (BMI) and waist-to-hip ratio (WHR). SUBJECTS: In total, 1241 Chinese women, who were recruited as community controls for a population-based case-control study of breast cancer. METHODS: Nineteen haplotype tagging single nucleotide polymorphisms (htSNPs) in four haplotype blocks were genotyped. RESULTS: Significant associations were observed for WHR at three SNPs that are located in haplotype block 1, including rs2445765, rs1004984 and rs1902584 (P=0.05, 0.04 and 0.01, respectively). Women, particularly premenopausal women, who carried the minor allele at any of these SNPs, had higher WHR than those without it. Of these three SNPs, the strongest association was observed at rs1902584, which is the closest to Promoter I.4, the major promoter for adipose tissue. Haplotype analyses indicated an association between the haplotype TCCAT in block 1 and WHR with a P-value of 0.02. CONCLUSION: These results suggested that CYP19A1 genetic polymorphisms may be associated with the risk of obesity among Chinese women, especially among premenopausal women. The CYP19A1 protein (aromatase) plays a critical role in estrogen biosynthesis and thus affects body fat distribution and regulation.

Population dynamics of anaerobic microbial consortia in thermophilic methanogenic sludge treating paper-containing solid waste.

To maintain a stable thermophilic (55 degrees C) anaerobic digestion treating toilet paper-containing garbage, it is necessary to operate the digester at long hydraulic retention time (HRT) and low organic loading rate (OLR). Critical conditions of the digestion were investigated by operating the digester at HRT 23 days and OLR 3.4 gCOD(Cr)/L/d (R1) or HRT 14 days and OLR 5.6 gCOD/Cr)/L/d (R2) separately. Characteristics of methanogenesis of the two digesters were examined by measuring gas generation volume and volatile fatty acids (VFA) concentration, and the populations of four anaerobic acidogens and three methanogens were analyzed using quantitative PCR method. In digester R1, methanogenic activity was unstable but it could be recovered by stopping feeding as though VFA accumulation occurred. The population of acidogens and two methanogens were maintained at 10(11) - 10(13) copies/L, however, the population of Methanoculleus could not be recovered after methanogenesis recovering. In digester R2, the period of methanogenesis was significantly shorter than that in digester R1. Both the acidogens and the methanogens could not be maintained at a stable concentration. It is suggested that the critical HRT to sustain the population of acidogens in this process should be longer than 14 days and for all kinds of methanogens, HRT should be longer than 23 days.

Field test of methane fermentation system for treating swine wastes.

A methane fermentation system for treating swine wastes was developed and successfully demonstrated in a field test plant (0.5 m3/d). The system was composed of a screw-press dehydrator, a methanogenic digester, a sludge separator, an oxidation ditch (OD) and composting equipment. A performance evaluation was carried out regarding physical pre-treatment using the screw-press dehydrator, methane fermentation for pre-treated slurry, and post-treatment for digested effluent by OD. Total solids (TS) and chemical oxygen demand (CODCr) removal by the screw-press pre-treatment were 38% and 22%, respectively. Properties of the screenings were as follows: water content 57%, ignition loss 93%, specific gravity 0.33. The pretreated strong slurry was digested under mesophilic conditions. Digestion gas (biogas) production rate was 25 m3/m3-slurry (NTP) and methane content of the biogas was 67%. CODCr removal of 65% with methane fermentation treatment of the slurry operating at 35 degrees C was observed. No inhibition of methane fermentation reaction occurred at the NH4(+)-N concentration of 3,000 mg/l or less during methane fermentation by the system. Mass balance from the present pilot-scale study showed that 1 m3 of mixture of excrement and urine of swine waste (TS 90 kg/m3) was biologically converted to 25 m3/m3-slurry (NTP) of biogas (methane content 67%), 100 kg of compost (water content 40%, ignition loss 75%), and 0.80 m3 of treated water (SS 30-70 mg/l).

Magoh, a human homolog of Drosophila mago nashi protein, is a component of the splicing-dependent exon-exon junction complex.

The RNA-binding protein Y14 binds preferentially to mRNAs produced by splicing and is a component of a multiprotein complex that assembles approximately 20 nucleotides upstream of exon-exon junctions. This complex probably has important functions in post-splicing events including nuclear export and nonsense-mediated decay of mRNA. We show that Y14 binds to two previously reported components, Aly/REF and RNPS1, and to the mRNA export factor TAP. Moreover, we identified magoh, a human homolog of the Drosophila mago nashi gene product, as a novel component of the complex. Magoh binds avidly and directly to Y14 and TAP, but not to other known components of the complex, and is found in Y14-containing mRNPs in vivo. Importantly, magoh also binds to mRNAs produced by splicing upstream (approximately 20 nucleotides) of exon- exon junctions and its binding to mRNA persists after export. These experiments thus reveal specific protein-protein interactions among the proteins of the splicing-dependent mRNP complex and suggest an important role for the highly evolutionarily conserved magoh protein in this complex.

SMN interacts with a novel family of hnRNP and spliceosomal proteins.

Spinal muscular atrophy (SMA) is a common neurodegenerative disease caused by deletion or loss-of-function mutations of the survival of motor neurons (SMN) protein. SMN is in a complex with several proteins, including Gemin2, Gemin3 and Gemin4, and it plays important roles in small nuclear ribonucleoprotein (snRNP) biogenesis and in pre-mRNA splicing. Here, we characterize three new hnRNP proteins, collectively referred to as hnRNP Qs, which are derived from alternative splicing of a single gene. The hnRNP Q proteins interact with SMN, and the most common SMN mutant found in SMA patients is defective in its interactions with them. We further demonstrate that hnRNP Qs are required for efficient pre-mRNA splicing in vitro. The hnRNP Q proteins may provide a molecular link between the SMN complex and splicing.

Role of the nonsense-mediated decay factor hUpf3 in the splicing-dependent exon-exon junction complex.

Nonsense-mediated messenger RNA (mRNA) decay, or NMD, is a critical process of selective degradation of mRNAs that contain premature stop codons. NMD depends on both pre-mRNA splicing and translation, and it requires recognition of the position of stop codons relative to exon-exon junctions. A key factor in NMD is hUpf3, a mostly nuclear protein that shuttles between the nucleus and cytoplasm and interacts specifically with spliced mRNAs. We found that hUpf3 interacts with Y14, a component of post-splicing mRNA-protein (mRNP) complexes, and that hUpf3 is enriched in Y14-containing mRNP complexes. The mRNA export factors Aly/REF and TAP are also associated with nuclear hUpf3, indicating that hUpf3 is in mRNP complexes that are poised for nuclear export. Like Y14 and Aly/REF, hUpf3 binds to spliced mRNAs specifically ( approximately 20 nucleotides) upstream of exon-exon junctions. The splicing-dependent binding of hUpf3 to mRNAs before export, as part of the complex that assembles near exon-exon junctions, allows it to serve as a link between splicing and NMD in the cytoplasm.

Gastric pneumatosis associated with duodenal stenosis and malrotation.

Pneumatosis intestinalis, which is linear or cystic gas within the bowel wall, is usually found in premature babies in association with necrotizing enterocolitis. Gastric pneumatosis defined as intramural gas of the stomach is a rare sign during infancy. We report an infant with Down's syndrome and a duodenal web with obvious gastric pneumatosis.

[Outbreak of measles in a hospital and measures taken against hospital infection--evidence of cost and benefits].

In Japan, an isolated vaccine of measles is used because MMR vaccines have been suspended due to the frequent occurrence of aseptic meningitis after their use. It is administered only once with the cover rate having been approximately 70%. An outbreak of measles was experienced in eight of our health care workers (three doctors, three nurses and two clerks of our hospital) and in seven of our medical students, accompanying local outbreaks. Their condition was severe enough to require admission to our school hospital. One of medical students developed encephalitis, but he has recovered completely. The social cost including medical cost per worker was approximately yen 500,000 (about $4,500). We measured antibodies against measles, rubella, chickenpox and mumps using the ELISA assay in 1048 health care workers less than 40 years of age and 99 medical students before their clinical practice two or three months after the outbreak. The cost including the measuring of antibodies and vaccination for these workers and students was approximately yen 2,800,000 and it will be yen 700,000 for new workers each year. The negative rates for workers were 1.2% for measles, 8.8% for rubella, 10.0% for mumps and 2.0% for chickenpox and those for the medical students were 1.0%, 22.0%, 9.0% and 5.0%, respectively. Of the workers, 9.6% who had more than the defined very high tiaer were thought to have subclinical reinfection because they were non-symptomatic. The rates were 1.2% for rubella, 0.5% for chickenpox and 0.0% for mumps for more than the defined high titers among the workers, suggesting a risk of outbreaks in the future. Therefore, the antibodies of health care workers and medical students should be measured at the hospital and the antibody-negative person should be vaccinated to prevent infection not only from their patients but also prevent other patients from becoming infected by contracted health care workers or students.

The Y14 protein communicates to the cytoplasm the position of exon-exon junctions.

We recently described an RNA-binding protein, Y14, that binds preferentially to spliced mRNAs and persists in the cytoplasm. Y14 is part of a multi-protein complex that also contains the mRNA export factor TAP. This suggests that splicing imprints the mRNA with a unique set of proteins that communicate the history of the transcript to the cytoplasm. Here, using microinjection of pre-mRNAs into Xenopus oocyte nuclei followed by immunoprecipitation of RNase-fragmented mRNAs from the cytoplasm, we show that Y14 is stably bound to sequences immediately upstream of exon-exon junctions. This feature appears to be unique to Y14. Using monoclonal antibodies that we produced against Aly/REF, another component recently reported to be an mRNA export factor, we show that Aly/REF is associated with spliced mRNAs in the nucleus but is not detectable on mRNAs in the cytoplasm. Thus, we propose that the splicing- dependent binding of Y14 provides a position-specific molecular memory that communicates to the cytoplasm the location of exon and intron boundaries. This novel mechanism may play an important role in post-splicing events.

[Comparison of positivity rates for antibodies against measles, rubella, chickenpox and mumps by assays].

Our previous study found mistakes by some doctors in the choice of an assay for determining antibodies in Japan. To compare the positivity rates for antibodies by assays, we measured the antibodies of measles, rubella, chickenpox and mumps from the same sera using such methods as the EIA, HI and CF assays. The subjects were 175 nursing students. The positivity rates for measles, chickenpox and mumps by the EIA assay were 96.6%, 93.7%, and 83.3%, respectively. Those for rubella by the HI and CF assays were 92.0% and 10.1%. The sensitivity rates for measles, chickenpox and mumps by the HI and CF assays, based on the results of the EIA assay, were 75.1%, 102.4% and 69.2% in the HI assay, and 20.6%, 38.7% and 8.0% in the CF assay, respectively. Our previous study showed that the sensitivity of the HI assay for rubella antibody is same as that of the EIA assay in Japan. Currently an EIA assay should be chosen for these antibodies and the HI assay or IAHA assay should be possible selections for rubella and chickenpox. However, international comparison of the cutoff titers for these antibodies should be considered.

Application of nitric oxide for a case of veno-occlusive disease after peripheral blood stem cell transplantation.

A 5-year-old girl at high risk for acute lymphoblastic leukemia was treated with high-dose chemotherapy and autologous peripheral blood stem cell transplantation (PBSCT). However, her condition was complicated by veno-occlusive disease of the liver (VOD) after PBSCT. For treatment of VOD, transdermal isosorbide tape was applied as a nitric oxide (NO) donor. The signs of VOD improved immediately after NO treatment was initiated, and the patient showed no side effects from the transdermal isosorbide tape.

Pre-mRNA splicing imprints mRNA in the nucleus with a novel RNA-binding protein that persists in the cytoplasm.

We describe a novel RNA binding protein, Y14, a predominantly nuclear nucleocytoplasmic shuttling protein. Interestingly, Y14 associates preferentially with mRNAs produced by splicing but not with pre-mRNAs, introns, or mRNAs produced from intronless cDNAs. Y14 associates with both nuclear mRNAs and newly exported cytoplasmic mRNAs. Splicing of a single intron is sufficient for Y14 association. Y14-containing nuclear complexes are different from general hnRNP complexes. They contain hnRNP proteins and several unique proteins including the mRNA export factor TAP. Thus, Y14 defines novel intermediates in the pathway of gene expression, postsplicing nuclear preexport mRNPs, and newly exported cytoplasmic mRNPs, whose composition is established by splicing. These findings suggest that pre-mRNA splicing imprints mRNA with a unique set of proteins that persists in the cytoplasm and thereby communicates the history of the transcript.

Natural language from function dynamics.

A new approach to syntax and semantics of language is presented as a form of function dynamics, which is studied both analytically and numerically. The iteration of the function dynamics leads to articulation and formation of rules, which depend on each other. A hierarchy of meta-rules as rules of rules also emerges through the iteration when the initial function is suitably embedded. Iteration of a model with dialogue between two function dynamics is shown to generate a higher level structure.

Low induction of varicella-zoster virus-specific secretory IgA antibody after vaccination.

Breakthrough after varicella vaccination occurs in approximately 2. 6% approximately 18.6% of immunocompetent children, but the reason has not been demonstrated clearly. As a first defense, specific secretory IgA antibody on the mucosa plays an important role in preventing invasion of microorganisms. To examine induction of varicella-zoster virus (VZV) specific secretory IgA after natural infection and vaccination and its booster mechanisms, 143 salivary samples were tested by ELISA. The VZV-secretory IgA values were significantly higher in the matched children after natural chickenpox than in those after vaccination, although the total secretory IgA did not differ between them. Two (7%) of the vaccinees lacked the sIgA antibody. In the elderly and in immunocompromised children, the VZV-secretory IgA values were no lower than those in healthy children, and they did not lack VZV-secretory IgA. The doctors and nurses taking care of patients with chickenpox had higher values than the other groups as did individuals who had had herpes zoster recently. VZV-secretory IgA was thought to be stimulated by exogenous and reactivated endogenous VZV to neutralize VZV with weak activity. These results suggest that low or no induction of VZV-secretory IgA antibody after vaccination may be one of the possible explanations for a breakthrough.

Nutritional assessment of peripheral blood stem cell transplantation in children.

Peripheral blood stem cell transplantation (PBSCT) has many advantages for patients because hematopoiesis and general condition return to normal more rapidly than they do following bone marrow transplantation. Thus, the authors hypothesize that the nutritional condition of patients also returns to normal more rapidly after PBSCT. The duration of insufficient nutrition was investigated in children undergoing PBSCT. The subjects of this study were 8 patients with malignant diseases. The factors measured were body weight, body fat, cholesterol, albumin, pre-albumin, and retinol-binding protein. These parameters were measured a day before transplantation, and then once a week for 4 weeks after transplantation. All parameters were recovered until day 28 from the lowest level in transplantation. In this study, all parameters returned to normal comparatively early. PBSCT causes little damage to patients' nutrition.

[Measures against hospital infection for students in medical schools in Japan--a survey by questionnaires].

We sent questionnaires to 80 medical schools in Japan to know what the problems in hospital infection for students in medical schools are. Seventy-one % were sent back to us. There have been hospital infection from patients to students in 12% of the medical schools, included 3 cases each of measles, chickenpox and mumps, 2 cases each of rubella, hepatitis B and tuberculosis. Fourteen % of the medical schools had reports about the past history of infection and vaccination from students, 70% of the schools determined their antibodies and the 28% did nothing. Ninety-three%, 25%, 23%, 18%, and 15%, of the schools determined antibodies for hepatitis B, rubella, measles, chickenpox and mumps, respectively. Some assays of the measurement were low in sensitivity. The cost for the determination was fully paid in 48% of the schools, but only partially supported in 35%. Tuberculin reaction was performed in 40% of the schools and then BCG was done in 57%. Vaccination was recommended in 40% of the schools. The cost of vaccination was all paid in 38% of them, which was only for hepatitis B, and partially supported in 15% of them.