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Background Chromosome 4q25 has been repeatedly identified as atrial fibrillation (AF)-sensitive locus in multiple genome-wide association studies (GWAS) and is considered to hold some clues to AF pathogenesis. We aimed to investigate the clinical utilities in Japanese and to unveil the function of the 4q25 locus in affecting transcription of adjacent genes. Methods We conducted AF GWAS in Japanese population (1382 AF cases and 1478 controls) and the replication panel (1666 AF cases and 1229 controls) with detailed clinical information which showed the acceleration of AF onset. Stepwise investigations with linkage disequilibrium analysis, histone code patterns, and reporter assay in the 4q25 locus were performed. Results The AF GWAS confirmed a significant association of rs4611994 and rs1906617 in chromosome 4q25 with AF. In the clinical analysis, AF onset of the individuals with risk allele accelerated 2.5 years compared with those with protective allele ( p = 0.00012). Next, in the functional analysis, three single nucleotide polymorphisms (SNPs) in the variant group selected by linkage disequilibrium analysis were identified as candidates for the cis -regulatory element toward adjacent genes in chromatin immunoprecipitation assay. Among them, rs4611994 and rs72900144 regions showed higher effects on the transcriptional activity of luciferase gene in the risk alleles than those in the protective alleles ( p < 0.0001, p < 0.005, respectively). Conclusions AF GWAS in Japanese confirmed the association with 4q25 locus and indicated that its SNP affected the acceleration of AF onset. The candidate regions of the causative SNPs, rs4611994 and rs72900144, could alter the adjacent gene expression level. [ABSTRACT FROM AUTHOR]

Abstract: A comparative study was performed to identify the microscopic characteristics of hair or feather in the five animal drug components contained in Shenrongbian pill. Penis et Testis Canis is 40±0.07 in the medulla index, with long circular, banana or triangular circular shaped medulla cells arranged in one line or network, and the hair cuticle is in imbrication (d, m) and flat wave (p) shape. Penis et Testis Equi is 66±0.10 in the medulla index, with ellipse, spindle or long strip-shaped medulla cells arranged in network, and the hair cuticle is in flat wave shape. Penis et Testis Bovis is 67±0.05 in the medulla index, with rectangle, spindle or polygon-shaped medulla cells arranged in ladder or network form, and the hair cuticle is in flat wave shape. Penis et Testis Mustelae is 29±0.05 in the medulla index, with ellipse-like, square-like or circular shaped medulla cells arranged in one line generally, and the hair cuticle is in acuminate (d, m), imbrication (m,p) and slightly flat wave (p) shape. Musculus et Bonis Passeris is 24±0.05 in the medulla index, with bamboo joint-shaped barbs and unclear medulla cells, without hair cuticle. [Copyright &y& Elsevier]

Abstract: The influence of quinone structures related to the formation of the “colloidal deposit” on Pb electrode immersed into water containing lignin preparations or pure organic compounds has been investigated. It was found that quinone structures in lignin preparations, as well as in pure organic compounds, play an important role in the formation of the “colloidal deposit”. [Copyright &y& Elsevier]

Gene targeting using short interfering RNA (siRNA) has become a common strategy to explore gene function because of its prominent efficacy and specificity. For the application of siRNA technology to gene therapy, however, still more efficient transduction of siRNA into target cells is needed. In this study, we developed an adenoviral vector harboring a tandem-type siRNA expression unit, in which sense and antisense strands composing the siRNA duplex were separately transcribed by two human U6 promoters. Targeting survivin, an antiapoptotic molecule widely overexpressed in malignancies but not detected in terminally differentiated adult tissues, this type of adenoviral vector (Adv-siSurv) successfully exerted a gene knockdown effect and induced apoptosis in HeLa, U251, and MCF-7 cells. These cancer cells, once infected with Adv-siSurv, displayed remarkably attenuated growth potential, both in vitro and in vivo. Moreover, intratumoral injection of Adv-siSurv significantly suppressed tumor growth in a xenograft model using U251 glioma cells. This novel modality may be a promising tool for cancer therapy. [Copyright &y& Elsevier]

Cadherin molecules are essential for tissue morphogenesis and are also related to cancer invasion and metastasis. Although normal melanocytes express E- and P-cadherin, the activity and expression of E- and P-cadherin in melanoma cells are still unknown. We measured the homophilic adhesion activity of human normal epidermal melanocytes and the melanoma cell lines MeWo and A375. The melanoma cells showed stronger homophilic adhesion activity than did the melanocytes, despite the lower expression of E- and P-cadherin in the melanoma cells. This result suggested that melanoma cells expressed other types of homophilic adhesion molecules. Using degenerate primers to amplify multiple cadherin subtypes, we performed a polymerase chain reaction (PCR) with the first strand of cDNAs generated by reverse transcription of the mRNAs of the melanoma cells, and we isolated two known cadherin fragments, N-cadherin and PC42, and six novel cadherin fragments, cadherins ME1-ME6. The reverse transcriptase-PCR. using specific primers of cadherins including E-, P-, and N-cadherins, PC42, and cadherins ME1-M.E6 revealed that the melanoma cells expressed more kinds of cadherin molecules than did the melanocytes. Such cadherins may play an important role in melanoma cell-cell adhesion. [ABSTRACT FROM AUTHOR]

Epidermolysis bullosa acquisita is a subepidermal blistering disease in which patients have autoantibodies against the non-collagenous domain of type VII collagen. Starting with previously isolated 1-kilobase pair (Kb) cDNA for this au- toantigen, we isolated overlapping cDNAs with a combined open reading frame of -&SIM;3.2 Kb, encoding most (&SIM;115 kilo- daltons [KDa]) of the N-terminal non-collagenous domain of type VII collagen. To localize immunogenic domains, we produced maltose-binding fusion proteins with cDNA encoding different portions of this autoantigen. These cDNA fragments scan from 5' to 3' of this non-collagenous domain and overlap each other. An immunoblot analysis of these fusion proteins with eight epidermolysis bullosa acquisita patient sera demonstrated that each patient serum binds to different regions of this molecule and that epitopes for these patient sera locate throughout this autoantigen. These data suggest that multiple epitopes on the N-terminal non-collagenous domain of type VII collagen are recognized by circulating autoantibodies in patients with epidermolysis bullosa acquisita. [ABSTRACT FROM AUTHOR]

The various clinical features of adult T-cell leukemia/lymphoma (ATL) are frequently accompanied by skin eruptions. Recently, a cutaneous type of ATL has been proposed by clinical studies. We analyzed the viral integration of human T-cell leukemia virus-I (HTLV-I) and monoclonal rearrangement of T-cell receptor (TCR) gene in blood lymphocytes and the cutaneous infiltrated cells of nine ATL patients with various clinical features and skin eruptions. We classified them by the results of Southern blot analysis and propose a cutaneous-type ATL accordingly. In two of them, we could detect the monoclonal integration of HTLV-I and T-cell monoclonality only in the skin but not in the peripheral lymphocytes. We also demonstrated the time course study in one patient. Clinicians should be aware of the HTLV-I positive cutaneous T cell lymphoma that can be named cutaneous-type ATL. Examination of viral integration and T-cell monoclonality in skin lesions is required to make an exact diagnosis of cutaneous ATL. [ABSTRACT FROM AUTHOR]

A partial cDNA clone (called BP cDNA) with coding sequences for the carboxy-terminal region of bullous pemphigoid (BP) antigen has been recently isolated and sequenced. In order to determine whether specific peptides encoded by the cDNA could be used to raise antibodies against BP antigen, fusion proteins derived from fragments of the BP cDNA and 17-mer or 19-mer synthetic peptides, corresponding to its deduced amino acid sequence, were used to generate rabbit antibodies. Three restriction enzyme fragments, 1179 bp (5' end), 264 bp (middle), and 546 bp (3' end), of the 1992 open reading frame (ORF) of BP cDNA were subcloned in frame into pEX plasmids to make β-galactosidase fusion proteins FP1, FP2, and FP3, respectively. Fusion proteins of the predicted molecular weight, and which bound anti-β-galactosidase antibodies, were produced, confirming the length of the predicted ORF. Rabbits immunized with FP1, but not FP3, produced antibodies, similar to authentic antibodies from BP patients, which: 1) bound the epidermal basement membrane at titers over 10,000, as determined by indirect immunofluorescence; 2) bound the basement membrane on the roof of 1 M NaCl-split skin; 3) immunoprecipitated the 230-kD BP antigen; and 4) bound the hemidesmosome, as determined by immunoelectron microscopy. Rabbits immunized with FP2 also produced lower titer BP-like antibodies. We further showed that short hydrophilic synthetic peptides, contained in FP1, could induce similar BP-like antibodies in rabbits at immunofluorescence titers up to 2560. These rabbit antibodies should prove useful for further studies on the function and structure of particular epitopes of BP antigen as well as on the pathophysiology of disease. [ABSTRACT FROM AUTHOR]

T-cell populations of 22 plaque lesions from seven mycosis fungoides patients were studied for clonal rearrangement of the β chain of the T-cell receptor (Tβ gene. All plaque lesions employed in this study showed clinically similar appearance. Histologically, all the biopsy specimens showed epidermotropism and the dermal infiltration of mononuclear cells including atypical cells. Histochemically, the majority of the infiltrated cells had surface markers of helper T cells. DNA extracted from skin lesions, peripheral lymphocytes, and lymph nodes revealed that the monoclonal expansion of T cells was different among patients and lesions. DNA extracted from the two skin lesions of case 1 revealed a clonal expansion of T cells. The rearranged bands persisted for about 1 year. In contrast, all lesions from casts 3-7 showed no rearranged band. Interestingly, three lesions from case 2 showed mixed type results, i.e., the monoclonal expansion of T cells was detected in one lesion but not in the other two lesions. Time course study of case 2 revealed that the same rearranged hand became detectable in all three skin lesions and a lymph node about 1 year later. These results suggest that in plaque lesions of mycosis fungoides, there are various stages of detectable monoclonality of infiltrating cells, although the clinical appearance of the plaques was similar, and that the variety of monoclonality may reflect the long clinical course of this disease. [ABSTRACT FROM AUTHOR]

The effects of substance P and substance K, which are coexpressed in the same mRNA as a β-preprotachykinin in peripheral tissues and released in the inflammatory lesion of the skin, were examined on epidermal proliferation using spontaneously transformed mouse epidermal cell line (Pam 212 cells). Substance P stimulated the synthesis of DNA of Pam 212 cells in the medium containing 2%- 10% fetal calf serum (FCS). Stimulation of DNA synthesis was dose dependent if the cells were cultured in the medium containing 2% FCS (quiescent condition). This effect was inhibited by spantide. In a serum-free medium, substance P had no effect on keratinocyte proliferation. In contrast, substance K, which shares a common amino acid sequence with substance P on its C-terminal, did not affect DNA synthesis of Pam 212 cells in either medium condition. Substance P released in inflammation may stimulate epidermal proliferation. [ABSTRACT FROM AUTHOR]

Background: Studies have reported associations between neck pain and degenerative changes in the cervical spine in women, and between neck pain and obesity or metabolic syndrome. The present study investigated associations between neck pain or stiffness and lipid profiles in subjects recruited from the general population of a Japanese community.Methods: The 1122 volunteers who participated in this study included 426 men (age 52.6 ± 15.5 years; body mass index 23.6 ± 3.2) and 696 women (age 55.3 ± 15.3 years; body mass index 22.2 ± 3.4). Each subject filled out a questionnaire about any neck pain or neck-shoulder stiffness experienced in the previous 3 months. We recorded the following laboratory results related to metabolic factors, including lipid profiles: total cholesterol, LDL and HDL cholesterol, triglycerides, free fatty acids, glucose, and hemoglobin A1c. We conducted logistic and linear regression analyses using the prevalence of neck pain or neck-shoulder stiffness as the dependent variable and age, sex, body mass index, and laboratory data as independent variables.Results: There was no significant difference in the prevalence of neck pain between men (20.5%) and women (21.5%). However, the prevalence of neck and shoulder stiffness was significantly higher in women (60.3%) than in men (38.0%; P < 0.05). Logistic and linear regression analyses showed a significant negative correlation between the prevalence of neck pain and LDL cholesterol (odds ratio [OR], 0.958; 95% CI, 0.921-0.997), and between the prevalence of neck-shoulder stiffness and age (OR, 1.025; 95% CI, 1.013-1.038) and gender (OR, 0.362; 95% CI, 0.25-0.494).Conclusion: LDL cholesterol was correlated with neck pain in this cross-sectional population-based study. [ABSTRACT FROM AUTHOR]

IntroductionAt present, adenoviral vectors have been widely used for gene transfer into tumor cells primarily because they can be produced in high titers and can infect many different cell types. However, a major problem when utilizing the adenoviral vectors for gene therapy applications is thought to be related to low transduction efficiency due to weak expression of the adenovirus receptor, coxsackie– adenovirus receptor (CAR) in cancer cells, or side effects due to expression of CAR in normal cells. There is an urgent requirement to improve the specificity of gene delivery via adenoviral vectors in the context of cancer gene therapy.Experimental designWe constructed a fiber modified Ad5 vector (Adv-FZ33) in which a IgG-binding domain (Z33) derived from staphylococcal protein A was inserted into the HI loop of knob protein. We then evaluated on both in vitro and in vivo levels the extent of retargeting towards and therapeutic effectiveness against CEA-positive gastric cancers when using the fully human CEA antibody (C2-45) complex with this modified vector.ResultsIn vitro LacZ or EGFP reporter gene expression after infection with Adv-FZ33 vectors conjugated to anti-CEA mAb was approximately 20 times higher than that obtained with vector conjugated to the control mAb (human IgG4). We generated Ax3CAUP-FZ33, which is an Adv-FZ33 derivative vector expressing a therapeutic gene, namely E. coli uracil phosphoribosyltransferase (UPRT) which converts 5FU directly to 5-fluorouridine monophosphate. When conjugated with the anti-CEA mAb, C2-45, Ax3CAUP-FZ33 enhanced the cytotoxicity of 5FU (19.7-fold in terms of IC50 values) against gastric cancer cells. In a nude mouse peritoneal dissemination model, use of Ax3CAUP- FZ33 conjugated with anti-CEA mAb C2-45, in combination with 5FU administration, resulted in greatly enhanced anti-tumor effects against CEA-positive cancer cells. Tumor growth in mice treated with Ax3UP-FZ33/C2-45/5FU was significantly suppressed, and tumor volumes were less than a quarter of those of the control IgG4 group (total weight, 0.12 ± 0.11 g; P < 0.05). Furthermore, we examined the survival effects of Ax3UP-FZ33 conjugated with anti- CEA mAb plus 5FU in mice with peritoneal disseminated gastric cancer. The median survival time of the Ax3CAUP-FZ33/C2-45/5FU group was significantly longer than those of the PBS and 5FU only treatment groups (p<0.01, versus PBS and 5FU only groups). On the other hand, no significant difference was present between the Ax3CAUP-FZ33/5FU or Ax3CAUP-FZ33/IgG4/5FU groups and the PBS or 5FU only groups.ConclusionsThese data suggest that CEA-targeted FZ33-mutant adenovirus- mediated gene delivery offers a strong and selective therapeutic modality against CEA-producing cancers. This novel strategy may be a promising tool for cancer gene therapy.Molecular Therapy (2006) 13, S114–S115; doi: 10.1016/j.ymthe.2006.08.355 [ABSTRACT FROM AUTHOR]

The microstructure of rapidly solidified melt-spun ribbons of (Fe 0.75 M 0.10 B 0.15 ) 100− x Cu x (M = Si, Zr) alloys was investigated focusing on amorphous-phase formation and the solidification structure. In this study, Fe–Cu–Si–B and Fe–Cu–Zr–B alloys were designed to show amorphous-phase formation and liquid-phase separation simultaneously. Amorphous-phase formation was confirmed in both Fe–Cu–Si–B and Fe–Cu–Zr–B alloys. Minor exceptions in a combination map of mixing enthalpy and quaternary predicted phase diagram are acceptable range for designing a quaternary Fe–Cu-based alloy system that shows liquid-phase separation in Fe-based and Cu-based liquids and the formation of an Fe-based amorphous phase. [ABSTRACT FROM AUTHOR]

The microstructure of nanoglobules with a core–shell structure embedded in an Fe–Zr–B-based amorphous phase by liquid phase separation was investigated in an Fe–Cu–Zr–B immiscible alloy. Transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), and scanning transmission electron microscopy (STEM) studies of the melt-spun ribbon clarified that an fcc Cu nanocrystalline core was covered by a B-enriched Fe-based amorphous shell in the nanoglobules. [ABSTRACT FROM AUTHOR]

The microstructure of arc-melted ingots and rapidly solidified melt-spun ribbons of quaternary Fe–Cu–Nb–B immiscible alloys was investigated, with a focus on amorphous-phase formation and solidification structure. A continuous melt-spun ribbon with an Fe–Nb–B-based amorphous matrix and 10–100 nm diameter fcc -Cu crystalline globules was obtained for the (Fe 0.75 Nb 0.10 B 0.15 ) 80 Cu 20 alloy. [ABSTRACT FROM AUTHOR]

Background: Ossification of the posterior longitudinal ligaments (OPLL) causes myelopathy with progression or trauma. Most OPLL patients visit the hospital after severe symptoms occur, and surgery did not supply complete relief in severe symptoms related to OPLL. While it is necessary to diagnose OPLL early and observe carefully, the nature of symptoms and asymptomatic OPLL have been unclear. The purposes of this study were to investigate the prevalence and symptoms of ossification of the posterior longitudinal ligaments (OPLL) of the cervical spine in the general population. Methods: The subjects were 1291 Japanese general residents. Radiographic OPLL was detected by lateral view of the cervical spine, and subjects were classified into OPLL and non-OPLL groups. Visual analog scales of neck stiffness, neck pain, arm pain, and the Japanese Orthopaedic Association Cervical Myelopathy Evaluation Questionnaire were examined. Neck and arm symptoms were compared among the presence of OPLL, and among subtypes (continuous, segmental, and mixed types) statistically. Results: Prevalence of OPLL was 3.7 % in overall participants, 3.9 % in symptomatic participants, and 2.2 % in asymptomatic participants. While 86.7 % of participants with OPLL had symptoms, the consultation rate was only 6.7 %. The female OPLL group had higher neck pain than the non-OPLL group, and neck function of JOACMEQ in the continuous type of OPLL was significantly restricted compared to those without OPLL and with the mixed type of OPLL. Conclusions: Prevalence of OPLL was 3.7 %, and most of them had neck and arm symptoms; however, they did not visit hospitals. Asymptomatic OPLL had the potential risk for spinal cord injury and myelopathy. Early detection and careful observation would prevent the severe symptoms induced by OPLL. [ABSTRACT FROM AUTHOR]

Abstract: Solvation/desolvation and the solid electrolyte interphase (SEI) formation at a graphite electrode during the initial charging process were investigated using in situ Fourier transform infrared spectroscopy (FTIR) measurements. These measurements were developed by applying a diamond attenuated total reflectance (ATR) crystal, which probed the electrolyte solvents at the surface of the graphite electrode and provided successive FTIR spectra with high signal-to-noise ratio. The charging process was performed in the Li(reference)/electrolyte/graphite(working)/Cu cell at a voltage ranging from 3.2 to 0.0001 V vs. Li/Li+. The measurement elucidated the change in the chemical bond of the electrolyte solvents. In an early stage, the amounts of solvated and desolvated solvents changed, providing evidence that the Li+ ions were intercalated into the graphite layer. The formation of the Li alkyl carbonate that forms the SEI layer was facilitated toward the end of the charging process. Measurements were also obtained of the electrolyte with a vinylene carbonate additive, and the contribution of the additive to the electrolyte solvent reduction was investigated. [Copyright &y& Elsevier]

Activation of the Wnt-signaling pathway is known to play a crucial role in carcinogenesis of various human organs including the colon, liver, prostate, and endometrium. To investigate the mechanisms underlying hepatocellular carcinogenesis, we attempted to identify genes regulated by β-catenin/Tcf complex in a human hepatoma cell line, HepG2, in which an activated form of β-catenin is expressed. By means of cDNA microarray, we isolated a novel human gene, termed MARKL1 (MAP/microtubule affinity-regulating kinase-like 1), whose expression was downregulated in response to decreased Tcf/LEF1 activity. The transcript expressed in liver consisted of 3529 nucleotides that contained an open reading frame of 2256 nucleotides, encoding 752 amino acids homologous to human MARK3 (MAP/microtubule affinity-regulating kinase 3). Expression levels of MARKL1 were markedly elevated in eight of nine HCCs in which nuclear accumulation of β-catenin were observed, which may suggest that MARKL1 plays some role in hepatocellular carcinogenesis. Neoplasia (2001) 3, 4–9. [ABSTRACT FROM AUTHOR]

Darier's disease (DD) is an autosomal dominant genodermatosis characterized by epidermal acantholysis and dyskeratosis. We have performed genetic linkage studies in 10 families with DD (34 affected) by analyzing 14 polymorphic microsatellite markers. Our suits confirm recent reports mapping the DD gene to chromosome 12q23-q24.l. Haplotype analysis of recombinant chromosomes in our families, along with previously reported data, narrow the location of the DD gene to a 5 cM interval flanked by the loci D12S354 and D12S84/D12S105. This localization allowed exclusion of two known genes, PLA2A and PAH, as candidate loci for DD. Three other gene loci (PPP1C, PMCH, PMCA1), mapping in 12q21-q24, remain potential candidates. [ABSTRACT FROM AUTHOR]

This study was performed to elucidate whether xeroderma pigmentosum complementation group A (XPA) carrier has DNA repair abnormality against sun-exposure and ultraviolet (UV)-mimetic chemical carcinogen 4-nitroquinoline 1- oxide (4NQO). Here we report three sporadic cases of XP that were defined as group A by genetic complementation test as well as polymerase chain reaction (PCR) analysis to detect the point mutation in the responsible gene for XPA. DNA repair analyses in the skin fibroblasts revealed that the cells from the patients were much more sensitive to UV and 4NQO and had extremely low UV-induced unscheduled DNA synthesis (UDS) than control cells, whereas the cells from the carriers (heterozygotes of XP) had sensitivity to UV and 4NQO and levels of UV-induced UDS similar to normal cells. These results indicate that the obligate heterozygotes, despite having a mutated allele in XPA complementing gene demonstrated by PCR, have no DNA repair abnormality after UV irradiation and UV-mimetic 4NQO treatment. Our observations imply that XPA heterozygotes do not have higher risk of skin cancers than normal subjects based on their DNA repair abnormality. [ABSTRACT FROM AUTHOR]

Background Cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) is the standard treatment regimen for malignant lymphoma. CHOP-induced neutropenia is a common clinical complication that often develops into life-threatening febrile neutropenia (FN). The incidence rate of FN in malignant lymphoma patients receiving a CHOP + rituximab regimen is approximately 10%–20% in clinical trials. Assessing FN-related factors in an individual hospital is an important step towards ensuring appropriate treatment. Therefore, we aimed to evaluate the incidence, risk factors, and clinical features related to FN in our institution and to determine the rate of treatment success in FN patients receiving antibiotics. Methods FN-related characteristics were investigated in 182 malignant lymphoma patients who were treated with the CHOP regimen between January 2013 and December 2018. All data were retrospectively collected from the electronic medical record system. Categorical factors were compared using the Fisher's exact test and Mann-Whitney U test. Logistic regression analysis was used to analyze FN-related risk factors. Results Among 182 patients, 79 (43.4%) were men. Median age was 67 years. Most patients (90.7%) had an Eastern Cooperative Oncology Group performance status (PS) of 0–2. We noted 80 FN episodes in 60 patients (33.0%). On multivariate analysis, lack of granulocyte colony stimulating factor (G-CSF) prophylaxis in cycle 1 and poor PS were significantly associated with FN occurrence. Documented infections were observed in case of 35 FN episodes (43.8%), and median fever duration was 3 days. Treatment was successful without any modifications in case of 64 episodes (80%). No deaths were observed. Conclusion FN incidence was 33.3% at our institution. Lack of G-CSF prophylaxis and poor performance status were risk factors for FN in this setting. Use of G-CSF prophylaxis in patients with poor PS may help in decreasing FN incidence. [ABSTRACT FROM AUTHOR]