Oral ribavirin for treatment of respiratory syncitial virus and parainfluenza 3 virus infections post allogeneic haematopoietic stem cell transplantation.

The prognosis for patients with respiratory syncytial virus (RSV) or parainfluenza virus type 3 (PIV3) respiratory tract infection post allogeneic haematopoietic progenitor cell transplant (HPCT) is historically poor. The use of oral ribavirin (RBV) has not been widely studied in this patient population. We examined the outcomes of 15 consecutive patients (RSV, n=13 and PIV3, n=2) treated with oral RBV post HPCT. Oral RBV was commenced at a starting dose of 10 mg/kg/day, increasing to a maximum dose of 60 mg/kg/day depending on response and tolerance. At diagnosis, seven patients had upper respiratory tract infection (URTI) and eight had lower respiratory tract infection (LRTI). The starting RBV dose of 10 mg/kg/day did not prevent the progression of URTI to LRTI in any patient. However, with dose escalation, six of the seven patients responded to RBV therapy and survived their infective episode. Of the eight patients presenting with LRTI, six patients survived their infection, again after dose escalation of RBV. There was no dose-limiting toxicity seen in any patient. Our results indicate that oral RBV has clinical efficacy in the treatment of RSV/PIV3 infection post HPCT. However, a starting dose of 10 mg/kg/day appears ineffective; we recommend a starting dose of 20 mg/kg/day in this patient group.

[Intramolecular homologous recombination event occurred in the spider egg case silk gene CySp2 of wasp spider].

To gain further understanding of egg case silk proteins gene family, Zhao et al. isolated two full-length cDNAs for egg case silk proteins, cylindrical silk protein 1 (CySpl) and cylindrical silk protein 2 (CySp2), from the wasp spider, Argiope bruennichi. CySp2 was reported to contain no apparent signal peptide sequences, and the CySp1-CySp2 complex, which would possess a signal peptide, would be transported across the endoplasmic reticulum and secreted to the Golgi. Genomic DNA sequencing is one approach that can be successfully utilized to retrieve 5' ends of silk genes; using this method, we retrieved the 5' end of CySp2. We found that CySp2 contained a typical signal peptide similar to that found in CySp1; thus, due to technical limitations, an artificial error had occurred in the CySp2 sequence reported by Zhao et al.

New and highly efficient expression systems for expressing selectively foreign protein in the silk glands of transgenic silkworm.

We constructed three different fibroin H-chain expression systems to estimate the efficacy of producing recombinant proteins in the cocoon of transgenic silkworms. The results showed that the three different EGFP/H-chain fusion genes were all expressed selectively in the posterior silk gland of the transgenic silkworm. The recombinant protein content of transgenic silkworm cocoons is up to 15% (w/w) when using the most highly efficient H-chain expression system. To our knowledge, in comparison with silkworm silk gland expression systems in the literature, the highly efficient expression system developed in this study is the most efficient silkworm silk gland expression system to date. This expression system is the best candidate for foreign gene production and for creation of novel functional silk material. The results suggested the N-terminal domain and the intron of the H-chain gene are important in the secretion of fibroin and its transcription, respectively.

Role of hypertension, dyslipidemia and diabetes mellitus in the development of coronary atherosclerosis in Japan.

The present study evaluated the effect of hypertension (HT), dyslipidemia and diabetes mellitus (DM) on the development of coronary atherosclerosis in the Japanese population, using a cross-sectional study of 433 patients (254 men and 179 women) aged 30 years or older who underwent coronary angiography for suspected or known coronary heart disease angina at 5 cardiology departments in the Fukuoka area between September 1996 and August 1997. Patients with a disease duration of 6 months or more were excluded. The main outcome measure was angiographically defined coronary artery stenosis and was found to a significant degree in 146 patients (33.7%). HT, DM, low levels of high-density lipoprotein cholesterol (HDL-C) and hypertriglyceridemia remained as significant coronary artery disease (CAD) risk factors even after controlling for age, sex, hospital, smoking, alcohol use, body mass index and leisure time physical activity. However, hypercholesterolemia was not a significant risk factor after adjusting for these variables. After controlling for these variables, DM, low HDL-C and hypertriglyceridemia were significant CAD risk factors for men, but only DM was a significant CAD risk factor in women. These results indicate that in Japan DM, low HDL-C and hypertriglyceridemia may be more important CAD risk factors than hypercholesterolemia.

A covariant change of the two highly conserved bases in the GTPase-associated center of 28 S rRNA in silkworms and other moths.

The GTPase-associated center in 23/28 S rRNA is one of the most conserved functional domains throughout all organisms. We detected a unique sequence of this domain in Bombyx mori species in which the bases at positions 1094 and 1098 (numbering from Escherichia coli 23 S rRNA) are C and G instead of the otherwise universally conserved bases U and A, respectively. These changes were also observed in four other species of moths, but not in organisms other than the moths. Characteristics of the B. mori rRNA domain were investigated by native polyacrylamide gel electrophoresis using RNA fragments containing residues 1030-1128. Although two bands of protein-free RNA appeared on gel, they shifted to a single band when bound to Bombyx ribosomal proteins Bm-L12 and Bm-P complex, equivalent to E. coli L11 and L8, respectively. Bombyx RNA showed lower binding capacity than rat RNA for the ribosomal proteins and anti-28 S autoantibody, specific for a folded structure of the eukaryotic GTPase-associated domain. When the C(1094)/G(1098) bases in Bombyx RNA were replaced by the conserved U/A bases, the protein-free RNA migrated as a single band, and the complex formation with Bm-L12, Bm-P complex, and anti-28 S autoantibody was comparable to that of rat RNA. The results suggest that the GTPase-associated domain of moth-type insects has a labile structural feature that is caused by an unusual covariant change of the U(1094)/A(1098) bases to C/G.

Analysis of proteins encoded in the bipartite genome of a new type of parvo-like virus isolated from silkworm - structural protein with DNA polymerase motif.

Bombyx mori densonucleosis virus type 2 (BmDNV-2) is a small, spherical virus containing two complementary single-stranded linear DNA molecules (VD1, VD2). BmDNV-2 is a new type of virus with a unique, yet unspecified replication mechanism which is different from that of parvoviruses (Bando, H., Choi, H., Ito, Y., Nakagaki, M. , Kawase, S., 1992. Structural analysis on the single-stranded genomic DNAs of the virus newly isolated from silkworm: the DNA molecules share a common terminal sequence, Arch. Virol. 124, 187-193; Bando, H., Hayakawa, T., Asano, S., Sahara, K., Nakagaki, M. , Iizuka, T., 1995. Analysis of the genetic information of a DNA segment of a new virus from silkworm, Arch. Virol., 140, 1147-1155; Hayakawa, T., Asano, S., Sahara, K., Iizuka, T., Bando, H., 1997. Detection of replicative intermediate with closed terminus of Bombyx densonucleosis virus. Arch. Virol. 142, 1-7). Recent analyses on the genomic information of BmDNV-2 identified open reading frames which code for three tentative nonstructural proteins and four (VP1 to 4) of the six known structural proteins (Bando, H., Hayakawa, T., Asano, S., Sahara, K., Nakagaki, M., Iizuka, T., 1995. Analysis of the genetic information of a DNA segment of a new virus from silkworm, Arch. Virol., 140, 1147-1155; Nakagaki et al., in preparation). In this report we demonstrate that the two largest ORFs, VD1-ORF1 and VD2-ORF1, code for the two remaining structural proteins. In addition, computer-assisted analysis revealed that the structural protein encoded in VD1-ORF1 contains sequences conserved among various DNA polymerases, and showed an evolutionary relationship with the DNA polymerases involved in protein-primed replication.

Temperature change of the lamellar structure of DPPC/disaccharide/water systems with low water content.

Temperature change in l-alpha-dipalmitoyl phosphatidylcholine (DPPC)/disaccharide systems with low water content (less than 8 wt. %) was investigated using X-ray diffraction within a range of two transition temperatures. X-ray diffraction above the higher transition temperature showed a broad symmetric peak, indicating the Lalpha phase. Below the higher transition temperature, two overlapping diffraction peaks were observed. After peak separation, temperature change in these systems was analyzed using peak parameters of the two peaks. Peak parameters of the lower angle peak changed continuously up to and above the higher transition temperature, suggesting the systems to be in a liquid crystal phase below the higher transition temperature. Fourier-transform infrared (FT-IR) spectra of the DPPC/trehalose system with 5.5 wt.% water showed the wave number of asymmetric stretching of phosphate groups to change at the lower transition temperature and that of symmetric stretching of CH2 groups, to change between the lower and higher transition temperatures. Thus, below the lower transition temperature, the system is shown to be in a gel phase. Conformational change in phosphate groups occurred at the lower transition temperature. Within the lower and higher transition temperatures, two phases were found to coexist and transition from the gel phase to Lalpha phase to occur continuously. Above the higher transition temperature, the system is in the Lalpha phase.

Effect of water on lamellar structure of DPPC/sugar systems.

The ability of two monosaccharides, four disaccharides and one trisaccharide, to lower the transition temperature of L-alpha-dipalmitoyl phosphatidylcholine (DPPC) was investigated using differential scanning calorimetry (DSC) and the ability of these sugars to change the lateral packing of the acyl chains of DPPC was investigated using wide-angle X-ray diffraction. The sugars affected the gel-liquid crystal transition temperature (Tc) of DPPC when the water content of the DPPC/sugar systems was less than 20 wt.%. Specifically, Tc of the DPPC without sugar increased to approximately 106 degrees C, the Tc of the DPPC/monosaccharide system remained almost constant at 43 degrees C and of the DPPC/disaccharide or trisaccharide systems decreased to approximately 24 degrees C. In the dehydrated state, di- and trisaccharides caused looser packing of the DPPC hydrocarbon chains than the monosaccharides did, and the sugars affected the packing mode in different ways. The addition of water caused this difference in the sugars' effects on the packing mode to disappear and further addition of water caused the effect of sugar to almost disappear. Thus, the addition of water to a DPPC/sugar system weakens the interaction between the sugar and lipid and strengthens the DPPC chain packing.

Analysis of the genetic information of a DNA segment of a new virus from silkworm.

In 1983, a parvo-like virus (Yamanashi isolate) was newly isolated from silkworm. However, unlike parvovirus, two DNA molecules (VD1 and 2) were always extracted from purified virions. To investigate the structure and organization of the virus genomes, we determined the complete nucleotide sequence of VD2. The sequence consisted of 6031 nucleotides (nts) and contained a large open reading frame (ORF1) with 3513 nts. A smaller open reading frame (ORF2) with 702 nts was found in the complementary sequence. Computer analysis revealed that both ORFs did not code for the major structural proteins (VP1, 2, 3, and 4). These results suggest that VD2 has not enough information to produce progeny virions by itself. Further, the structural importance of the terminal sequence (CTS) common to both VD1 and VD2 was also predicted by a computer analysis.

Ionic size and behavior of diclofenac salts in water and ethanol/water mixtures by conductivity at 25 degrees C.

The objectives of this study were to evaluate the size of the diclofenac ion and the electrical behavior of diclofenac salts (DM; M = Li, Na, or K) in ethanol/water mixtures. The molar conductance of DM in aqueous solution was measured in a dilute concentration range of < 10 mol.m-3 at 25 degrees C. The Fuoss-Onsager conductance equation for unassociated electrolytes was used to determine the limiting molar conductance (lambda degrees) of DM. The molar conductivity of DM at a finite concentration was, however, larger than the value calculated by the Fuoss-Onsager limiting law. To correct the difference in the experimental data, two equations were employed to fit the experimental molar conductance of DM. The size of the diclofenac ion was calculated from the limiting molar conductance and compared with the values obtained by the Fedors method and the apparent ionization constant (pKa) of sodium diclofenac in ethanol/water mixture. The Walden products of DM increased up to 20% (w/w) ethanol, but above this concentration of ethanol they decreased. Specific interactions between the ions and solvent are discussed.

Retroperitoneal fibrosis associated with scirrhous gastric cancer.

A case of retroperitoneal fibrosis associated with scirrhous gastric cancer is reported. A sixty two-year-old Japanese female was admitted because of acute renal failure. The patient's serum creatinine level showed 3.2 mg/dl while the blood urea nitrogen level was 23 mg/dl. An ultrasound study of the upper abdomen revealed bilateral hydronephrosis. Drip infusion pyelography revealed a dilated right renal pelvis without ureteral obstruction. The left kidney was not opacified, suggesting a functional disorder. Gastrography and gastrofiberscopy revealed scirrhous gastric cancer. Signet ring cell carcinoma was later demonstrated histologically by biopsy specimens. CT demonstrated a prominent thickening of the gastric wall and hydronephrosis, although no prevertebral soft tissue masses were observed. A total gastrectomy was performed with failure to surgically decompress the ureters because fibrous plaque had firmly enveloped the retroperitoneal structures. Biopsy specimens of the retroperitoneum revealed an invasion of the tumor cells and prominent fibrosis. As an etiology of renal failure, ureteral stenosis resulting from secondary retroperitoneal fibrosis was also considered.

Non-specific activity of (+/-)-CP-96,345 in models of pain and inflammation.

The non-peptide NK1 receptor antagonist, CP-96,345, and its 2R,3R enantiomer CP-96,344, which is not an NK1 receptor antagonist (IC50 > 10 microM), were evaluated for antinociceptive and anti-inflammatory activities in several classical models of pain and inflammation in the rat. Both CP-96,345 and CP-96,344 reduced carrageenin-induced paw oedema and hyperalgesia, and attenuated the second phase of formalin-induced paw licking with equal potency. These results indicate that NK1 antagonism is not responsible for the activity of (+/-)-CP-96,345 in the above animal models.

Permeability of insulin entrapped in liposome through the nasal mucosa of rabbits.

The permeability of liposome entrapping insulin through the nasal mucosa of rabbit has been studied and compared with the permeability of insulin solution with or without pretreatment by sodium glycocholate (GC). Insulin entrapped in liposome was not detected in the receiver cell using the diffusion cells with the nasal mucosa. On the other hand, permeability of insulin entrapped in liposome increased after the pretreatment of GC. The phospholipids which result from liposomes, were not observed in the receiver. Also, the GC remaining in the nasal mucosa was measured. Considering the mechanism of permeation of insulin entrapped in liposome through the nasal mucosa, the GC remaining in the nasal mucosa may cause the lysis of liposomes.

Monolayer-bilayer equilibrium of phospholipid: stabilization of neutral lipid droplets in aqueous medium and catabolism of plasma lipoproteins.

Phospholipid spreads as monolayer at air/water and oil/water interfaces. Interfacial pressure of the monolayer in equilibrium with a definite bulk phase of the lipid, (equilibrium) spreading pressure, depends on lyotropic and thermotropic polymorphic state of the lipid bulk phase. Phosphatidylcholine (PC) in hydrated liquid crystalline state (bilayers in L alpha state) gives an appreciably large value of spreading pressure, 45-46 mN/m. Monolayer-bilayer equilibrium of a neutral lipid-phospholipid mixture is determined by miscibilities of the lipids in mixed monolayer, PC bilayer and bulk (liquid or solid) phase of the neutral lipid. A neutral lipid of limited solubility in PC bilayer forms a separate phase in aqueous medium. The phase is stabilized as small particles in the medium by PC monolayer at the particle surface. The monolayer is in equilibrium with the bilayer. This sort of equilibrium plays important roles in formation and catabolism of triglyceride- and cholesteryl ester-rich lipoprotein particles in animal plasma. The equilibrium is a critical factor also in stabilization of aqueous dispersion of lipophilic vitamin (neutral lipid). Coexistence of emulsion particles (neutral lipid core covered with PC monolayer) and vesicles made of PC bilayer are observed in a stable dispersion.