Comparing the risk of hepatitis B virus reactivation between direct-acting antiviral therapies and interferon-based therapies for hepatitis C.

Hepatitis B virus (HBV) reactivation has been reported during antihepatitis C treatment in patients with hepatitis C virus (HCV) and HBV co-infection. We aimed to evaluate the frequency and risk factors of HBV reactivation during anti-HCV therapy and compared those between interferon (IFN)-free direct-acting antiviral (DAA) therapies and IFN-based therapies. Three hundred and twenty-two patients with HCV infection receiving anti-HCV therapy were retrospectively screened. The baseline HBV infection statuses of all eligible patients and the HBV-DNA level of all patients with current or previous HBV infection were examined at the end of treatment. In patients with baseline anti-HBs positivity, changes in anti-HBs titre were evaluated. Of 287 patients who met the inclusion criteria, 157 had current (n=4) or previous (n=153) HBV infection; 85 were treated with IFN-free DAA therapies and 72 were treated with IFN-based therapies. Six patients experienced HBV reactivation (n=2) or HBV reappearance (n=4) after IFN-free DAA therapies, while no patient developed HBV reactivation after IFN-based therapies. The risk factors of HBV reactivation or reappearance were DAA therapies and a reduction in anti-HBs titre to <12 mIU mL-1 by the end of treatment. The decline changes of anti-HBs titre were significantly higher in patients treated with DAA therapies. Although HBV reactivation hepatitis was not observed, three of four patients with HBV reactivation or reappearance after achieving HCV eradication had viremia 8 weeks after completion of therapy. A significant proportion of patients develop HBV reactivation or reappearance without hepatitis after IFN-free DAA therapies. Low levels of anti-HBs and their decrease to <12 mIU mL-1 after treatment are significant risk factors for HBV reactivation or reappearance.

Genomic structure and inducible expression of the IL-22 receptor alpha chain in mice.

IL-22 is a newly identified member of the interferon/IL-10 family. In humans, IL-22 signals through a heteroduplex receptor consisting of IL-22R and CRF2-4/IL-10Rbeta. To investigate the physiological function of IL-22 and IL-22R, we isolated a cDNA encoding the mouse IL-22R, which has been a missing component of the functional receptor complex for mouse IL-22. Subsequently, we identified the genomic sequence of the mouse IL-22R gene by a database search. The gene consists of about 24 kb and is split into seven exons. Interestingly, intron 2 begins with a GC dinucleotide instead of the consensus GT, although otherwise the overall structure of the mouse IL-22R gene is strikingly similar to its human counterpart. The gene was mapped to mouse chromosome 4 in the region syntenic to the human IL-22R gene locus. In normal mice, IL-22R mRNA is detected at very low levels in restricted organs such as the kidney, liver, and lung. However, upon lipopolysaccharide stimulation, IL-22R mRNA expression is highly upregulated in the liver, in contrast to CRF2-4, which is expressed constitutively in a variety of tissues. Thus, the expression of the functional IL-22 receptor in the liver is regulated at the gene transcription level.

Overexpression of IL-15 in vivo enhances protection against Mycobacterium bovis bacillus Calmette-Guérin infection via augmentation of NK and T cytotoxic 1 responses.

To investigate the immunomodulating effects of IL-15 in vivo on mycobacterial infection, we used IL-15-transgenic (Tg) mice, which were recently constructed with cDNA-encoding secretable isoform of IL-15 precursor protein under the control of a MHC class I promoter. The IL-15-Tg mice exhibited resistance against infection with Mycobacterium bovis bacillus Calmette-Guérin (BCG), as assessed by bacteria growth. IFN-gamma level in serum was significantly higher in IL-15-Tg mice than in non-Tg mice after BCG infection. NK cells were remarkably increased, and Ag-specific T cytotoxic 1 response mediated by CD8+ T cells producing IFN-gamma was significantly augmented in the IL-15-Tg mice following BCG infection. Neutralization of endogenous IFN-gamma by in vivo administration of anti-IFN-gamma mAb deteriorated the clearance of the bacteria. Depletion of of NK cells or CD8+ T cells by in vivo administration of anti-asialo-GM(1) Ab or anti-CD8 mAb hampered the exclusion of bacteria. Thus, overexpression of IL-15 in vivo enhanced protection against BCG infection via augmentation of NK and T cytotoxic 1 responses.

Impaired IL-15 production associated with susceptibility of murine AIDS to mycobacterial infection.

LP-BM5 murine leukemia virus (MuLV) injection causes murine AIDS (MAIDS), a disease characterized by many functional abnormalities of immunocompetent cells. We show that MAIDS mice are susceptible to Mycobacterium bovis Bacille Calmette-Guérin (BCG) infection as assessed by survival rate and bacterial counts. The peritoneal exudate macrophages from MAIDS mice produced a significant level of interleukin (IL)-12 soon after inoculation with BCG, whereas IL-15 and tumor necrosis factor (TNF) production were severely impaired in BCG-infected MAIDS mice. The appearance of natural killer (NK) and CD4+ T helper type 1 (Th1) cells specific for mycobacterial antigen were depressed in MAIDS mice after BCG infection. Thus, it appeared that impaired production of IL-15, besides other inflammatory cytokines, in MAIDS mice may be involved in the poor responses of the NK and Th1 cells, resulting in an increased susceptibility to BCG.

Expression of mouse mammary tumor virus superantigen accelerates tumorigenicity of myeloma cells.

To investigate whether superantigen (SAG) from endogenous mouse mammary tumor virus functions as an immunogenic or a tumorigenic factor in tumor development, the BALB/c myeloma cell line FO was transfected with the SAG gene from the 3' Mtv-50 long terminal repeat (LTR) open reading frame (ORF), the product of which was specific for Vbeta6. All five transfectants expressing Mtv-50 LTR ORF mRNA showed stimulatory activity for Vbeta6 T-cell hybridomas in vitro; this activity was inhibited by the addition of anti-Mtv-7 monoclonal antibody (MAb) or anti-major histocompatibility complex class II I-A(d) and I-E(d) MAb. All transfectants with the SAG gene grew more rapidly than did mock transfectants in BALB/c mice after subcutaneous inoculation, whereas all clones, including mock transfectants, grew equally well in athymic nude mice. A significant fraction of Vbeta6 T cells selectively expressed activation markers, including CD44(high), CD62L(low), and CD69(high), and produced large amounts of interleukin 5 (IL-5) and IL-6 in BALB/c mice inoculated with transfectants. These results suggested that the expression of viral SAG enhances the tumorigenicity of a myeloma cell line through the stimulation of SAG-reactive T cells.

Differential roles of interleukin 15 mRNA isoforms generated by alternative splicing in immune responses in vivo.

At least two types of interleukin (IL)-15 mRNA isoforms are generated by alternative splicing at the 5' upstream of exon 5 in mice. To elucidate the potential roles of IL-15 isoforms in immune responses in vivo, we constructed two groups of transgenic mice using originally described IL-15 cDNA with a normal exon 5 (normal IL-15 transgenic [Tg] mice) and IL-15 cDNA with an alternative exon 5 (alternative IL-15 Tg mice) under the control of an MHC class I promoter. Normal IL-15 Tg mice constitutionally produced a significant level of IL-15 protein and had markedly increased numbers of memory type (CD44(high) Ly6C(+)) of CD8(+) T cells in the LN. These mice showed resistance to Salmonella infection accompanied by the enhanced interferon (IFN)-gamma production, but depletion of CD8(+) T cells exaggerated the bacterial growth, suggesting that the IL-15-dependent CD8(+) T cells with a memory phenotype may serve to protect against Salmonella infection in normal IL-15 Tg mice. On the other hand, a large amount of intracellular IL-15 protein was detected but hardly secreted extracellularly in alternative IL-15 Tg mice. Although most of the T cells developed normally in the alternative IL-15 Tg mice, they showed impaired IFN-gamma production upon TCR engagement. The alternative IL-15 transgenic mice were susceptible to Salmonella accompanied by impaired production of endogenous IL-15 and IFN-gamma. Thus, two groups of IL-15 Tg mice may provide information concerning the different roles of IL-15 isoforms in the immune system in vivo.

Vestibular and cochlear toxicity of aminoglycosides--a review.

Recently, there have been many reports describing the efficacy of intratympanic aminoglycoside injection for the treatment of intractable vertigo in patients with Ménière's disease. However, the number of injections and the amount of drug injected varies, with concomitant variation in the side-effect of hearing deterioration. To identify drugs that are more selectively vestibulotoxic, we have reviewed the ototoxicity of aminoglycosides, focusing on differences between vestibulo- and cochleotoxicity. At present, the basis for the different effects of each drug is unknown. The mechanisms of vestibulo- and cochleotoxicity are deemed worthy of further study.

Influence of music on heart rate variability and comfort--a consideration through comparison of music and noise.

By considering three kinds of music and noise, this research investigates the influence of music on the living body by comparing the difference of influence on heart rate variability and comfort when subjects listen to music and are exposed to noise. We used two pieces of classical music, rock music, and noise recorded by a tape recorder. The following conclusions were made from the findings of the research: 1) Hearing classical music results in a small variance of Mayer Wave related Sinus Arrhythmia (MWSA) component and Respiratory Sinus Arrhythmia (RSA) component compared with a body being at rest. This is because the sympathetic nerve is suppressed by the sound of classical music. With rock music and noise, however, the MWSA component increases and the RSA component decreases. 2) From a psychological evaluation, we found that classical music tends to cause comfort and rock music and noise tend to cause discomfort. 3) A correlation was found between the balance of the MWSA component and the RSA component and the psychological evaluation. As the comfort increases, the variance of MWSA decreases; as discomfort increases, the variance of MWSA increases.

Quantitative determination of cardiac glycosides in Digitalis lanata leaves by reversed-phase thin-layer chromatography.

An analytical method for the determination of cardiac glycosides in Digitalis lanata leaves by reversed-phase thin-layer chromatography (RP-TLC) was developed. The procedure consisted of extraction of dry leaf powder with 50% methanol and clean-up by Sep-Pak cartridges prior to RP-TLC analysis. RP-TLC was performed on an octadecylsilyl bonded silica gel plate, using a developing solvent of acetonitrile-methanol-0.5 M NaCl (1:1:1) for primary glycosides and acetonitrile-methanol-0.5 M NaCl (12:7:9) for secondary glycosides. The plate was scanned with a reflectance densitometer at 225 nm. The quantitation was carried out by the internal standard method. The present method is reliable and relatively simple for the determination of cardiac glycosides in Digitalis lanata leaves.

Age-related changes in learning and memory and cholinergic neuronal function in senescence accelerated mice (SAM).

The senescence-accelerated mouse (SAM) has been established as a murine model of accelerated aging. We investigated learning ability and memory in various tasks in a SAM strain, SAMP1TA, and in a control strain of SAMR1TA at the ages of 20, 30 and 40 weeks. We also measured choline acetyltransferase (ChAT) and cholinesterase (ChE) activity in the brains of these mice at the same ages. In a Y-maze task, in which short-term memory can be examined, there was no difference in learning ability between SAMP1TA and SAMR1TA at any age. Ability in latent learning and passive-avoidance tasks was less in SAMP1TA at 30 weeks of age than in age-matched SAMR1TA. The level of ChAT activity in the striatum of SAMP1TA was lower, than that of SAMR1TA at the ages of 20 and 30 weeks. At the ages of 40 and 50 weeks, ChE activity in the striatum of SAMP1TA was lower than that of SAMR1TA. These results suggest that SAMP1TA has a deficit, with cholinergic neuronal dysfunction, in learning ability and memory, as shown by impairment of performance in latent learning and long-term memory, but not in short-term memory.

Characteristics of Roman character input in Japanese word processing on keystroke interval time.

This study clarifies the characteristics of the time keys that are taken to enter Roman letters for Japanese sentences. First, test the appearance frequency of two consecutive characters of the alphabet in the input of English sentences and in the input of the Roman characters of Japanese sentences. Based on these results, we analyzed the features of stroke frequency of the keys in the Roman character input. Based on the result of the analysis, we clarified the features of the keystroke interval time for two consecutive characters in the Roman character input. Then, keys were struck to enter Japanese sentences for a fixed period of time and a test was made for the characteristics. As a result, the following points were clarified: 1) The time keys struck were divided into two large groups. One for two consecutive alphabet letters that corresponded to Kana, or the Japanese syllabary, and the other with no correspondence. The former had smaller values for the mean time and deviations, and the latter had larger values for them. 2) The subjects who struck a larger number of keys per unit time took less time in typing two consecutive letters which did not correspond to Kana.

Osteoinductive activity of composites of bone morphogenetic protein and pure titanium.

Titanium sponges were infused with bone morphogenetic protein (BMP-Ti), and the osteoinductivity of the resultant composite was measured. New bone formation occurred three weeks after implantation and was identified by soft x-ray analysis. Quantitative analysis showed no significant difference between BMP-Ti composites and control samples (BMP only). Consequently, pure titanium neither inhibited nor promoted BMP activity. Chondrocytes and new bone formation occurred in direct contact with the surfaces of the titanium. X-ray microanalysis demonstrated new bone formation inside the pores of the titanium sponges. The BMP-Ti composite has interesting properties as an osteoinductive implant and has potential practical clinical applications.

Influence of noise on heart rate and quantity of work in mental work.

In the present study, we investigated the correlation between the physiological function, heart rate (HR) and the quantity of work performed when A-weighted sound pressure level of factory noise or construction noise was varied during the mental operations of calculation and erosion. Furthermore, we looked for differences in the effects of sound noise and music on the quantity of work performed. The A-weighted sound pressure level was set at four levels of background noise, i.e. 60, 70 and 80 dB (A). The music sound level was set at 70 dB (A). The subjects were exposed to factory noise, construction noise and music during their mental work. A balanced factorial experiment was conducted with a total of 16 combinations of types of mental work (two levels of calculation and erosion), sound noise (two levels of factory and construction sound noise), and A-weighted sound pressure levels (four levels). The order of the experimental procedures was determined at random. The following conclusions were made from the findings of the study. 1) HR elevation occurred due to exposure to sound noise, and further increase in HR resulted from an increase in the A-weighted sound pressure level. This result can be attributed to the action of sound noise as a stressor, which excites sympathetic nerves, leading to an elevation of HR. 2) The quantity of work performed decreased with an increase in the A-weighted sound pressure level. 3) When the subjects were exposed to music with an A-weighted sound pressure level of 70 dB (A) instead of sound noise of the same level, almost no elevation of HR was observed. However, the quantity of work completed when exposed to music was less than that performed when exposed to sound noise of the same A-weighted sound pressure level.

[The treatment of condyle neck fracture: statistics gathered by multi centric study and the related prognosis].

There are two different treatments for condyle neck fracture. One is non-surgical treatment involving intermaxillary fixation, and the other is radical surgical treatment through various techniques, but it is difficult to discuss the relative merits. The purpose of this report is to decide whether surgical treatment is necessary, by comparing the two types of treatment. In the period between April 1981 to December 1987, we treated 136 cases of condyle neck fracture at the Second Department of Oral and Maxillofacial Surgery School of Dentistry Aichigakuin University, and in 18 nearby hospitals. Of the 136 cases, 58 cases had fracture only at the condyle neck, while 78 cases had some concomittant fracture. As regards treatment, of the 136 cases, 49 (36%) were treated surgically, while 87 (64%) were treated non-surgically for condyle neck fracture. The surgically treated cases were divided into three groups: 24 cases were treated by pinning, 16 cases were treated by inter osseous wiring, and there were 9 other cases. In the treatment of concomittant fracture, 38 cases involved inter osseous wiring, 22 cases involved metal plate fixations, and there were 4 other cases. Non-surgical treatment accounted for 15 cases. The rate of surgical treatment for concomittant fracture was 82.1% of the cases treated. In surgically treated cases of condyle neck fracture, the mouth opening prognosis was generally poor. However, in these cases the deviation of mouth opening was generally better than in non-surgically treated cases. There were no differences in mastication, occlusion and pain in the TMJ region.

Effect of diethyl-beta-cyclodextrin on the release of nitroglycerin from formulations.

The complex-forming abilities of 2,6-di-O-ethyl-beta-cyclodextrin (DE-beta-CD), and its effect on the release of nitroglycerin (TNG) from formulations of the compound, were studied and compared with corresponding properties of beta-cyclodextrin (beta-CD) and 2,6-di-O-methyl-beta-cyclodextrin (DM-beta-CD). Complex formation was confirmed by differential scanning calorimetry and infrared absorption spectroscopy. In an accelerator test involving temperature and reduced pressure, marked depression of the volatility of TNG was observed as a result of CD complex formation. Dissolution rates of TNG from powdery TNG/DE-beta-CD complex and its tablets were retarded in comparison with the rates from other CD complexes. The release rate of TNG from ointments was accelerated by complexation with DE-beta-CD, and retarded by complexation with beta-CD. To evaluate their in vivo percutaneous absorption, samples were applied to the inside tip of the cheek pouch of male golden hamsters. The amount of TNG remaining in the cheek pouch was lowest in the case of the TNG/DE-beta-CD complex ointment, and relatively high in the case of the TNG/beta-CD complex ointment, in agreement with the in vitro results. We suggest that the combination of DE-beta-CD complex and beta-CD complex might be applicable to sustained-release preparations for percutaneous administration.

[Bone inductive activity of hydroxyapatite-bone morphogenetic protein complex].

Bone morphogenetic protein (BMP) is known to be a protein which induces new bone at heterotopic sites. Purification of BMP has not been perfected, and obtaining large amounts of BMP is very difficult, so it seems better to use some carrier or frame material for BMP to work effectively. Various kinds of hydroxyapatite (HAP) have been used to repair periodontal osseous defects, but they do not have osteogenetic or osteoinductive properties. If osteoinductive proteins such as BMP could retain their biologic properties after being implanted into living tissue with HAP, it would be an advantage in repairing periodontal osseous defects. In this experiment, we prepared BMP-HAP complex and investigated its osteoinductive activity. BMP was extracted from bovine cortical bones in accordance with the Urist's procedure. The ability of this BMP to stimulate new bone growth was ensured by implantation in the muscle pouch of mice. HAP was synthesized by the wet method. The BMP-HAP complex was implanted in the muscle pouch of mice, and osteoinduction was examined 3, 7, 14, and 21 days after implantation to assess its osteo-inductive ability. New bone formation was studied by roentgenographic and histologic observation. In the BMP-HAP group, new bone formation was seen on the roentgenograms and new cartilage and bone were observed histologically in the tissue surrounding the apatite. In the HAP group, no new cartilage or bone formation was noted.

[Bone inductive activity of bone morphogenetic protein combined to beta-tricalcium phosphate].

For the development of a new delivery system for bone morphogenetic protein (BMP), BMP was bound to beta-tricalcium phosphate (beta-TCP). Powder beta-TCP was synthesized from calcium hydrogenphosphate and calcium carbonate by the dry process method to make the best use of the advantages of the BMP, which show good bone inductive activity on the surface and enhancement of new tissue by reducing the area that the implant material occupies. The beta-TCP + BMP complex and beta-TCP for the controls only were implanted in the muscle pouches of mice. Three weeks later new bone formation was observed on the exterior surface of beta-TCP + BMP complex but not of beta-TCP controls. The bone inductive activity of the beta-TCP + BMP complex is better than the BMP alone. The histological relation between the original tissue and the newly induced bone formation was normal and that of new bone and beta-TCP was also good. Consequently, the beta-TCP + BMP complex has good histocompatibility when implanted.