Association of cigarette smoking with a past history and incidence of herpes zoster in the general Japanese population: the SHEZ Study.

Few studies have examined the impact of cigarette smoking on the risk for herpes zoster. The Shozu Herpes Zoster (SHEZ) Study is a community-based prospective cohort study over 3 years in Japan aiming to clarify the incidence and predictive and immunological factors for herpes zoster. We investigated the associations of smoking status with past history and incidence of herpes zoster. A total of 12 351 participants provided valid information on smoking status and past history of herpes zoster at baseline survey. Smoking status was classified into three categories (current, former, never smoker), and if currently smoking, the number of cigarettes consumed per day was recorded. The participants were under the active surveillance for first-ever incident herpes zoster for 3 years. We used a logistic regression model for the cross-sectional study on the association between smoking status and past history of herpes zoster, and a Cox proportional hazards regression model for the cohort study on the association with risk of incidence. The multivariable adjusted odd ratios (95% CI) of past history of herpes zoster for current vs. never smokers were 0·67 (0·54-0·80) for total subjects, 0·72 (0·56-0·93) for men and 0·65 (0·44-0·96) for women. The multivariable adjusted hazard ratios (95% CI) of incident herpes zoster for current vs. never smokers were 0·52 (0·33-0·81) for total subjects, 0·49 (0·29-0·83) for men and 0·52 (0·19-1·39) for women. Smoking status was inversely associated with the prevalence and incidence of herpes zoster in the general population of men and women aged ⩾50 years.

Assessment of skin test with varicella-zoster virus antigen for predicting the risk of herpes zoster.

The Shozu Herpes Zoster (SHEZ) Study was designed to clarify the incidence of and predictive and immunological factors for herpes zoster in a defined community-based Japanese population. As part of this series, a total of 5683 residents aged ≥50 years received a varicella-zoster virus (VZV) skin test with VZV antigen, and 48 h later, the erythema and oedema were assessed by measuring the longest diameter. The diameters of both the erythema and oedema decreased with the increasing age of the subject. Sixty-three subjects contracted herpes zoster within a year after receiving the VZV skin test. Analysis of the herpes zoster incidence rate vs. the skin test reaction revealed that the shorter the diameter of erythema or oedema, the greater the likelihood of herpes zoster. These results demonstrated that the VZV skin test is an excellent surrogate marker for predicting the risk of herpes zoster.

Development of fiber-substituted adenovirus vectors containing foreign peptides in the adenovirus serotype 35 fiber knob.

Fiber-substituted adenovirus (Ad) vectors containing fibers of Ad serotype 35 (AdF35) efficiently transduce a variety of human cells because their receptor, human CD46, is ubiquitously expressed on almost all nucleated cells. However, the ubiquitous expression of CD46 might lead to unexpected transduction in untargeted organs. In this study, we developed fiber-modified AdF35 vectors with an integrin-binding Arg-Gly-Asn (RGD) peptide incorporated into the FG, HI or IJ loop, which have been identified as important regions for binding to CD46. Incorporation of foreign peptides into these loops does not inhibit trimerization of the fibers. In CD46-negative cells, fiber-mutant AdF35 vectors containing an RGD peptide in the FG or HI loop showed 6- to 30-fold higher transduction efficiencies in an RGD-peptide-dependent manner than the unmodified AdF35 vectors. In contrast, in CD46-positive cells, insertion of foreign peptides markedly reduced the transduction efficiencies of the AdF35 vectors, indicating that insertion of foreign peptides significantly inhibits binding to CD46. In particular, CD46-mediated transduction was completely diminished by insertion of foreign peptides into the HI loop. Our findings indicate that HI loop is the most suitable domain to mediate a foreign peptide-dependent and CD46-independent transduction by incorporation of foreign peptides into the Ad35 fiber knob.

IL-6-induced homodimerization of gp130 and associated activation of a tyrosine kinase.

The biological functions of interleukin-6 (IL-6) are mediated through a signal-transducing component of the IL-6 receptor, gp130, which is associated with the ligand-occupied IL-6 receptor (IL-6R) protein. Binding of IL-6 to IL-6R induced disulfide-linked homodimerization of gp130. Tyrosine kinase activity was associated with dimerized but not monomeric gp130 protein. Substitution of serine for proline residues 656 and 658 in the cytoplasmic motif abolished tyrosine kinase activation and cellular responses but not homodimerization of gp130. The IL-6-induced gp130 homodimer appears to be similar in function to the heterodimer formed between the leukemia inhibitory factor (LIF) receptor (LIFR) and gp130 in response to the LIF or ciliary neurotrophic factor (CNTF). Thus, a general first step in IL-6-related cytokine signaling may be the dimerization of signal-transducing molecules and activation of associated tyrosine kinases.

Relationship between U83 gene variation in human herpesvirus 6 and secretion of the U83 gene product.

The betaherpesvirus human herpesvirus 6 (HHV-6) has two variants. The U83 gene product of strain HST is a chemoattractant for monocytes. Here, we describe U83 gene variations that accumulated in variants A and B. A gene-variation hot spot was examined in 36 different strains and one donor DNA sample. U83 gene variations accumulated in variant A and in reactivated variant B after transplantation. None of the variant-A viruses encoded the signal peptide found in the B variant. U83 gene sequencing suggested that the variant A and B groups were separate, and that the variant B viruses could be further divided into the HST-Z29 type and another type with a shorter signal peptide. In a eukaryotic expression system, the HST-Z29 type of U83 gene product was secreted into the medium, a frame-shifted HST-Z29 type was partially secreted, and the variant-A type and a first-methionine knockout of the HST-Z29 type were not secreted.

Reactivation of latent human immunodeficiency virus 1 by human herpesvirus 6 infection.

Infection of the ACH-2 line of human leukemic T cells carrying latent Human immunodeficiency virus 1 (HIV-1) with Human herpesvirus 6 (HHV-6) resulted in an increase in reverse transcriptase (RT) activity, a marker of HIV-1 activation, in the culture supernatant. A similar effect was obtained with 12-O-tetradecanoyl-phorbol-13-acetate (TPA). The RT activity reached a peak at 24 hrs post infection (p.i.) and then declined, suggesting that the cells underwent lysis. The HIV-1 antigen was co-expressed with an early-late HHV-6 product, but not always with an immediate-early (IE) HHV-6 product, suggesting that one or more IE gene products were involved in the activation of latent HIV-1 in ACH-2 cells.

Preferential dilation of recipient coronary arteries of the collateral circulation by intracoronary administration of nitroglycerin.

OBJECTIVES: The purpose of this study was to test the hypothesis that the sensitivity to nitroglycerin of collateral vessels and recipient arteries is greater than that of donor arteries of the collateral circulation. BACKGROUND: The collateral circulation responds vigorously to nitroglycerin. However, the mechanisms of the efficacy of nitroglycerin for improving collateral circulation are not fully elucidated. METHODS: The diameter of donor and recipient arteries of the collateral circulation was measured with a computer-assisted analysis system in eight patients with well developed collateral vessels. Coronary angiography was repeated before and after the intracoronary injection of 50 micrograms of nitroglycerin. RESULTS: After nitroglycerin, the mean diameter +/- SD of donor arteries increased to 1.61 +/- 0.53 from 1.29 +/- 0.39 mm (p < 0.01), whereas the diameter of recipient arteries increased to 1.59 +/- 0.50 from 1.10 +/- 0.49 mm (p < 0.01). The change in the diameter of recipient arteries was significantly greater than that of donor arteries (52.3 +/- 24.6% vs. 24.7 +/- 11.5%, p < 0.05). These changes induced by the intracoronary injection of nitroglycerin were accompanied by a decrease in pacing-induced ST segment depression (0.16 +/- 0.06 to 0.06 +/- 0.04 mV, p < 0.01), suggesting increased flow reserve through collateral channels. CONCLUSIONS: These findings indicate that the sensitivity to nitroglycerin of recipient arteries of the collateral circulation is significantly greater than that of donor arteries. This observation may explain the strong response of the collateral circulation to nitroglycerin in patients with functionally significant collateral channels.

Relation between collateral channel filling and flow grade in recipient coronary arteries in patients with stable effort angina.

OBJECTIVES: The purpose of the present study was to elucidate the relation between the extent of perfusion of the ischemia-related coronary artery and the degree of visualization of the collateral circulation to the ischemic area. BACKGROUND: Because it is difficult to accurately assess coronary stenosis severity with standard angiographic techniques, the inclusion of flow grade in recipient coronary arteries would provide an additional perspective concerning the effect of the progression of atherosclerotic obstructive disease on the development of collateral circulation. METHODS: The coronary arteriograms of 54 consecutive patients with chronic effort angina without prior myocardial infarction were examined. Patients were classified into four groups according to the extent of perfusion of the ischemia-related coronary artery (Thrombolysis in Myocardial Infarction [TIMI] grade 0 to 3). The degree of angiographically demonstrable collateral circulation was also classified into four grades (collateral index 0 to 3). RESULTS: Eighteen patients had TIMI grade 0, 6 had grade 1, 13 had grade 2 and 17 had grade 3 perfusion. The collateral indexes of TIMI 0, 1, 2 and 3 groups were 2.7 +/- 0.7 (mean +/- SEM), 2.2 +/- 0.6, 1.2 +/- 1.1 and 0.4 +/- 0.9, respectively (p < 0.01 vs. TIMI 0, p < 0.05 vs. TIMI 1). CONCLUSIONS: These findings indicate that all patients with chronic effort angina have the potential for collateral development as a result of coronary artery narrowing, and the functional state of well developed collateral vessels may be primarily determined by the pressure gradient across the collateral network.

Establishment and characterization of the T-cell line, EBT-8 latently infected with Epstein-Barr virus from large granular lymphocyte leukemia.

An Epstein-Barr virus (EBV) genome-positive T-cell line, designated EBT-8, was established from peripheral blood of a patient with EBV genome-positive large granular lymphocyte leukemia of T-cell origin. The cells have been cultured continuously in RPMI-1640 medium supplemented with 10% fetal calf serum and 40 U/ml interleukin-2 for more than 18 months. Analysis of T-cell receptor gene rearrangement demonstrated similar rearrangement between the fresh leukemic cells and EBT-8 cell line. The cell line has several azurophilic granules in its cytoplasm and activated cytotoxic/suppressor T-cell surface antigens (CD2, CD3, CD8, HLA-DR and T-cell receptor alpha/beta). Karyotypic analysis of the cell line showed several chromosomal abnormalities. EBV DNA was demonstrated in the cells by Southern blot hybridization and about five copies of covalently closed circular DNA per cell were detected by Gardella gel analysis. Clonotypic episomal EBV DNA was observed in the cells by Southern blot hybridization with EBV-terminal fragment probe. EBV-encoded small RNA, EBER1 were demonstrated in all cells by in situ hybridization. EBV-encoded proteins, EBNA and LMP1 were demonstrated by immunofluorescence technique. EBV activation was observed after 12-O-tetradecanoylphorbol-13- acetate treatment of the cells. These results demonstrated the establishment of a T-cell line with latent EBV genomes and suggested the involvement of EBV to the large granular lymphocyte leukemia of T cells.

Transformation of large granular lymphocytic leukemia during the course of a reactivated human herpesvirus-6 infection.

A patient with CD3+ large granular lymphocytic (LGL) leukemia developed transformation (TF). The phenotype of the leukemic cells was CD3+, CD4+ and CD8-. The leukemic cell count increased rapidly; the cells became large and the nuclear outline, which had been reniform, became lobulated. Anti-HTLV-1 and anti-HIV antibodies were negative in the serum of the patient and no HTLV-1 specific sequences were detected in the cDNA of the leukemic cells by polymerase chain reaction (PCR). Comparison of the karyotype abnormality of the leukemic cells before and after TF revealed an abnormality of the 21 trisomy in 90% of mitotic cells of the patient. Analysis of the cell cycle revealed that 13.7% of the leukemic cells were in DNA synthesis phase which was not previously found. The titer of anti-human herpesvirus-6 (HHV-6) immunoglobulin G which had been high at chronic phase (1:1640 compared to normal titer of less than 1:160), became 1:20,000 at TF. The titer of anti-HHV-6 immunoglobulin M also increased from less than 1:4 at the chronic phase to 1:120 at TF (normal value less than 1:4). A HHV-6-specific DNA sequence was detected by PCR in the peripheral mononuclear cells collected at TF but not at the chronic phase. These data suggests that TF occurs not only in CD3-negative but also in CD3-positive LGL leukemia. HHV-6 reactivation is therefore a possible cause in immunocompromised hosts whose general conditions are deteriorated.

Importance of myocardial ischaemia for recruitment of coronary collateral circulation in dogs.

STUDY OBJECTIVE: The aim of the study was to investigate collateral coronary flow and regional myocardial function following different coronary occlusion protocols. DESIGN: Effects of brief left anterior descending artery (LAD) occlusions in dogs (using a pneumatic occluder around the proximal artery) on collateral circulation were evaluated using three different protocols, each producing the same period of pressure gradient across the collateral network: (1) 10 s occlusion X 30 at 1 min intervals; (2) 1 min occlusion X 5 at 1 min intervals; (3) 5 min occlusion X 1. Each protocol was followed by a 10 s occlusion after a further 1 min period. SUBJECTS: 14 mongrel dogs of either sex were used, weight 10-21 kg. MEASUREMENTS AND MAIN RESULTS: Left ventricular pressure, left circumflex coronary artery (LCCA) flow, and subendocardial segment shortening (% delta L) in the area perfused by the LAD were monitored. Collateral blood flow from LCCA to LAD territory was measured as a stepwise decrease in LCCA flow on release of LAD occlusion. During the first 10 s of occlusion, % delta L decreased from 23.6(SEM 2.2)% to 14.2(2.9)%. After protocol (1), % delta L decreased from 23.1(2.2)% to 14.8(3.0)%. By contrast, after protocol (2) and (3) % delta L decreased only slightly, from 22.7(2.6)% to 20.5(2.8)%, and from 22.4(2.4)% to 19.8(2.4)%, respectively. Although collateral blood flow remained unchanged after protocol (1), it increased from 1.6(0.4) ml.min-1 during the first LAD occlusion to 3.0(0.7) ml.min-1 (p less than 0.05) after protocol (2), and to 3.5(0.6) ml.min-1 (p less than 0.05) after protocol 3. Haemodynamic measurements prior to each 10 s LAD test occlusion remained unchanged throughout the experiment. CONCLUSIONS: The pressure gradient across the collateral network cannot dilate pre-existing collateral vessels by itself, but ischaemia related metabolites may play an important role in the recruitment of collateral circulation.

Up-regulation of elafin/SKALP gene expression in psoriatic epidermis.

The expression of mRNA for elafin/SKALP, an inhibitor of leukocyte elastase and proteinase 3, in human normal and psoriatic epidermis was examined by in situ hybridization. In normal epidermis, elafin/SKALP mRNA was detected in the granular layer, but not in the spinous or basal layers. In fully developed psoriatic lesions, elafin/SKALP mRNA was found in the suprabasal layers of the ridges, and in the upper two thirds of the stratum malpighii at the elongated rete ridges. Intense staining was noted near the subcorneal microabscess in psoriasis vulgaris and under the subcorneal pustule in localized pustular psoriasis. In the marginal psoriatic epidermis, elafin/SKALP mRNA was expressed from the middle or upper spinous layer to the subcorneal layer, and the cells expressing elafin/SKALP mRNA increased especially under the parakeratotic corneal layer intermingled with pyknotic nuclei of neutrophils. These findings suggest that the induction of elafin/SKALP gene expression is related closely to the infiltration of neutrophils into the epidermis in psoriasis and plays an important role in protecting the skin components against the tissue damage caused by the infiltrated leukocytes.

Cholesterol sulfate activates transcription of transglutaminase 1 gene in normal human keratinocytes.

Cholesterol sulfate and transglutaminase 1 are essential for the process of keratinization. Cholesterol sulfate is formed during keratinization and activates the eta isoform of protein kinase C. Transglutaminase 1 is a key enzyme for formation of the cornified envelope in terminally differentiated keratinocytes. In this study, we demonstrated that cholesterol sulfate acts as a transcriptional activator of the transglutaminase 1 gene in normal human keratinocytes. Growth of normal human keratinocytes was inhibited by cholesterol sulfate, but not by its parental cholesterol. Treatment of normal human keratinocytes with cholesterol sulfate induced activity of transglutaminase 1 in a dose- and time-dependent manner. Activation of transcription of transglutaminase 1 by cholesterol sulfate was demonstrated by northern blotting analysis, whereas that by cholesterol was not. In order to identify a cholesterol sulfate responsive region in the transglutaminase 1 gene, plasmids were constructed containing a luciferase reporter gene ligated to deletion fragments of the 5' upstream region of the tranglutaminase 1 gene and were transfected into normal human keratinocytes. Transfected cells were treated with cholesterol sulfate, the phorbol ester 12-O-tetradecanoylphorbol-13-acetate and a high concentration of Ca2+. Our results indicate that the responsive element(s) for cholesterol sulfate and phorbol ester is located upstream of the human transglutaminase 1 gene at a position(s) between -819 and -549, whereas the responsive element for Ca2+ is located at a position between -79 and -49.

Analysis of epidermal-type transglutaminase (TGase 3) expression in mouse tissues and cell lines.

In the formation of the cornified cell envelope in the epidermis, epidermal-type transglutaminase (TGase 3) cross-links a variety of structural proteins. However, its expression in other tissue has not been investigated. Furthermore, no cell line expressing TGase 3 has been found. The tissue distribution of TGase 3 in mice was investigated using reverse-transcription polymerase chain reaction (RT-PCR) and Western blotting analyses. TGase 3 mRNA was expressed in the brain, stomach, spleen, small intestine, testis, skeletal muscle and skin. The stomach and testis expressed TGase 3 protein in size similar to that observed in the epidermis. Screening various cell lines, a gastric human cancer cell line, MKN-1 and mouse neuroblast cell line, neuro2a, were found to express TGase 3.

Effects of vanadate on intracellular Ca2+ redistribution and hexose transport across plasma membrane in cultured mouse fibroblasts.

The effects of vanadate on intracellular Ca2+ sequestration and hexose transport were studied in Swiss 3T3 cells. Vanadate inhibited ATP-dependent Ca2+ uptake by saponin-permeabilized Swiss 3T3 cells at 10(-5) and 10(-7) M Ca2+ at which the Ca2+ uptake was sensitive and insensitive to oligomycin plus antimycin A, respectively. On the other hand, vanadate stimulated 2-deoxy-D-glucose (2DG) uptake in a dose- and time-dependent way. The stimulation of 2DG uptake by vanadate was inhibited by EGTA plus A23187 and the inhibition was reversed by Ca2+ restoration. These results suggest that an increase in cytosolic Ca2+ by inhibition of intracellular ATP-dependent Ca2+ sequestration by vanadate results in the stimulation of hexose transport in Swiss 3T3 cells.

Upstream activation element of the PH03 gene encoding for thiamine-repressible acid phosphatase in Saccharomyces cerevisiae.

The PH03 gene of Saccharomyces cerevisiae encodes thiamine-repressible acid phosphatase and requires the positively acting regulatory protein THI2 for its expression. Deletion analysis of the 5'-flanking region of PH03 gene revealed that an activating region located at nucleotide position -234 to -215 relative to the translation initiation codon is required for the expression and sensitivity to thiamine. A chemically synthesized DNA fragment covering -234 to -215 showed a significant level of expression when inserted in front of the PH03 promoter lacking the activating region. Electrophoretic mobility shift assay demonstrated the presence of proteins that bound to the above DNA fragment in the nuclear extract from cells grown in thiamine-free medium. These findings suggested that this region between -234 and -215 acts as an upstream activation element of the PH03 gene that can interact with regulatory proteins.

Defective terminal differentiation and hypoplasia of the epidermis in mice lacking the Fgf10 gene.

Here, we characterized the skin and hair phenotype of mice lacking the fibroblast growth factor 10 gene (Fgf10), a newly identified member of the fibroblast growth factor family. Histological examination of Fgf10(-/-) newborn mouse skin revealed abnormalities in epidermal morphogenesis. The number of proliferating cells in the basal layer was decreased, the granular layer was hypoplastic and lacked distinctive keratohyaline granules and tonofibrils. The expression of loricrin, a marker of epidermal differentiation, was dramatically reduced. Despite the presence of Fgf10 transcripts in normal hair follicles, abnormalities of hair development were not observed in Fgf10(-/-) skin. These data suggest that Fgf10 is required for embryonic epidermal morphogenesis but is not essential for hair follicle development.

Demonstration of Borna disease virus RNA in peripheral blood mononuclear cells derived from Japanese patients with chronic fatigue syndrome.

CFS, a recently named heterogeneous disorder, is an illness of unknown etiology. The association of CFS with viral infections has been suggested. A common association between CFS and several viruses examined has not been confirmed. Here, we centered on the possible link between CFS and BDV infection. By nested RT-PCR followed by hybridization, BDV RNA was demonstrated as a clear signal in PBMCs in 3 out of 25 CFS patients. The amplified cDNA fragments were cloned and sequenced. A total of 16 clones were studied. Intra-patients divergencies of the p24 were 2-9%, 3-20%, and 3-11% in the deduced amino acids. Inter-patient divergencies among the 16 clones were 3-24%. Antibodies to recombinant BDV p24 protein were detected in 6 CFS patients including one carrying BDV RNA. Overall, these gave the prevalence of 32% (8/25) in Japanese CFS patients, suggesting that Japanese CFS is highly associated with active infection of BDV, or a related agent.

Human herpesvirus 6 infection in renal transplantation.

The relationship between renal transplantation and human herpesvirus 6 (HHV-6) infection was studied. All 21 kidney donors examined had antibody to HHV-6 at the time of transplantation. The 21 kidney recipients also had detectable antibody to HHV-6 before transplantation--and, of these, 8 patients showed a significant increase of serum antibody titer against HHV-6 after transplantation. All these 8 recipients suffered severe kidney rejection. Furthermore, virus isolation from peripheral blood lymphocytes of 2 recipients who suffered rejection was attempted, and in both cases HHV-6 was isolated. Biopsy specimens of rejected kidneys of 9 other patients were examined for the presence of HHV-6 antigens, and in 5 of these specimens antigens were detected in the tubular epithelium, as well as in infiltrating histiocytes and lymphocytes. These results suggest that HHV-6 can infect renal tissues and that the infection may be correlated with rejection or with immunosuppressive therapy.