Changes in the composition of molecular species of covalently bound and free ceramides [EOS], and their correlation with disease severity in atopic dermatitis.

Ceramides are major constituents of stratum corneum (SC) intercellular lipids involved in skin barrier function. The ratio of molecular species of ceramides and their correlation with disease severity was examined in patients with atopic dermatitis (AD). Thirty-eight patients with AD and 32 healthy controls (HCs) were assessed for transepidermal water loss, SC collection and clinical assessment. The ceramide content of different molecular species in the samples was quantified using high-performance liquid chromatography coupled with tandem mass spectrometry. Unsaturated acyl chains of both covalently bound and free ceramides [EOS] were higher in AD lesional skin than those in AD non-lesional or normal HC skin. The proportion of unsaturated acyl chains (C30:1, C32:1 and C34:1) was higher than other ceramide molecular species among covalently bound and free ceramides [EOS] in patients with AD. The proportion of unsaturated acyl chains in covalently bound ceramides was positively correlated with transepidermal water loss (r = 0.600) when considering the total number of non-lesional and lesional skin. Additionally, thymus and activation-regulated chemokine (TARC) showed a positive correlation with unsaturated acyl chains proportion in AD non-lesional (r = 0.676) and lesional (r = 0.503) skin. Our study is the first to show the increase in unsaturated acyl chains of both covalently bound and free ceramides [EOS] in lesional and non-lesional skin in AD for each molecular species. This increase is associated with dryness and impaired barrier function, which correlates with TARC levels, a marker for the degree of type 2 inflammation. We speculate that type 2 inflammation exacerbation leads to abnormal epidermal lipid metabolism in the skin of patients with AD.

Light-induced displacement of a microbead through the thermal expansion of liquid crystals.

We found the displacement of a microbead by the oblique irradiation of UV light in pure 4-cyano-4'-pentylbiphenyl (5CB) without photo-responsive molecules. This system is very simple and a complicated experimental setup is not necessary. The displacement is observed in the nematic state of 5CB and not in the isotropic state. The distance of the displacement was proportional to the intensity of the UV light. After the irradiation was stopped, the microbead started to return to the original position. These phenomena are attributed to the thermal expansion of 5CB induced by the photo-thermal effect of polyimide coated on the glass substrate.

CD10 as a novel marker of therapeutic resistance and cancer stem cells in head and neck squamous cell carcinoma.

BACKGROUND: Cancer stem cells (CSCs) are responsible for treatment failure. However, their identification and roles in resistance are not well established in head and neck squamous cell carcinoma (HNSCC). METHODS: Three HNSCC cell lines (FaDu, Detroit562 and BICR6) were treated with cisplatin or radiation. Cell surface antigens were analysed by LyoPlate, a novel cell surface antigen array. The expression levels of antigens highly expressed after treatments were further compared between cisplatin-resistant Detroit562 cells and its parental line. Association of the candidate antigen with CSCs properties, namely sphere formation and in vivo tumourigenicity, was also examined. RESULTS: CD10, CD15s, CD146 and CD282 were upregulated across the treated cell lines, while the increased expression of CD10 was prominent in the cisplatin-resistant cell line. Isolation mediated by FACS revealed that the CD10-positive subpopulation was more refractory to cisplatin, fluorouracil and radiation than the CD10-negative subpopulation. It also showed an increased ability to form spheres in vitro and tumours in vivo. Moreover, the CD10-positive subpopulation expressed the CSC marker OCT3/4 at a higher level than that in the CD10-negative subpopulation. CONCLUSIONS: CD10 is associated with therapeutic resistance and CSC-like properties of HNSCC. CD10 may serve as a target molecule in the treatment of refractory HNSCC.

Dewetting of a droplet induced by the adsorption of surfactants on a glass substrate.

The dewetting of a surfactant droplet, induced by the adsorption of surfactants on a hydrophilic glass substrate, was observed experimentally. After being dropped onto the substrate, the droplet began shrinking and the speed of shrinkage increases with the surfactant concentration. We explained this dynamics, which is known as reactive dewetting, semi-quantitatively by expanding a simple theoretical model originally proposed and discussed qualitatively by M. E. R. Shanahan and P.-G. de Gennes [Start-up of a reactive droplet, C. R. Acad. Sci. Paris, 1997, 324, 261-268]. In addition, for the surfactant droplet with a concentration near or higher than the critical micelle concentration (CMC), we found that it maintains a large final contact area compared with that in the case of the low surfactant concentration. We discussed this phenomenon by taking the decrease in the vapor-liquid and solid-liquid interfacial tensions into consideration.

Effectiveness of the computed analysis of electroglottographic signals in muscle tension dysphonia.

OBJECTIVE: To investigate the usefulness of electroglottography (EGG) parameters in the diagnosis and estimation of efficacy of voice therapy for muscle tension dysphonia (MTD). PATIENTS AND METHODS: Nineteen MTD participants, an equivalent number of dysphonic ('organic') patients with vocal fold lesions and as many normal speakers were enrolled. Acoustic (Ac) and EGG signals during sustained phonation were recorded simultaneously. The period and amplitude perturbation quotient of both signals, the closed quotient (CQ) of EGG signals (mean CQ) and its standard deviation (CQSD) were calculated, and subsequently compared among the three groups. These parameters in the MTD group were compared before and after voice therapy. RESULTS: The perturbation measures of both signals in the MTD group were either as high as or significantly higher than those in the organic group or the control group, respectively. Both the Ac and EGG parameters after therapy significantly decreased. The CQSD, but not mean CQ, also decreased after therapy. CONCLUSION: EGG parameters related to the regularity of vocal fold vibration, but not to the degree of vocal fold contact (mean CQ), are useful for the diagnosis and estimation of voice therapy outcome in MTD.

pH-dependent Ca2+ interaction with phospholipids related to phosphatidylethanolamine N-methylation.

The interactions of PE and its N-methylated derivatives (PME, PDE AND PC) WITH Ca2+ were examined. PE and the intermediate phospholipids of PE N-methylation (PME and PDE) interacted with Ca2+ in a pH-dependent and reversible manner. When these phospholipids were present in the heptane phase, Ca2+ in the aqueous phase was translocated into the heptane phase at alkaline pH but not at acidic pH. PDE was also effective for the translocation even at around neutral pH, while PC hardly translocated Ca2+ at pH 6.0-9.2. The amounts of Ca2+ interacting with these phospholipids were in the following order: PDE is greater than PME is greater than PE is much greater than PC. P1, phosphatidic acid and PS interacted with Ca2+ in the whole pH range examined. The Ca2+ interactions with P1 and phosphatidic acid were independent of pH, while PS interacted with more Ca2+ at alkaline pH. These phospholipids interacted with Ca2+ most strongly among the cations studied. Liposomes containing PDE also bound the highest amounts Ca2+ among PE and its N-methylated derivatives. Furthermore, mammalian cultured cell membranes, which contain increased amounts of PDE by in vivo modification with N,N'-dimethylethanolamine, bound more Ca2+ than those prepared from choline-treated control cells.

N-glycosylation does not affect assembly and targeting of proglycinin in yeast.

Glycinin, a simple protein, and beta-conglycinin, a glycoprotein, are the dominant storage proteins of soybean and are suggested to be derived from a common ancestor. To investigate why glycinin does not require glycosylation for its maturation, we attempted N-glycosylation of proglycinin A1aB1b using site-directed mutagenesis and yeast expression system. An N-glycosylation consensus sequence Asn-X-Ser/Thr was created at positions 103, 183, 196, 284 and 457 in the variable regions being strongly hydrophilic revealed from the alignment of amino acid sequences of various glycinin-type proteins. Among five mutant proglycinins (Q103N, H183N, G198T, S284N, N459T), Q103N was fully glycosylated, and H183N and N459T were partly (around 20% of the expressed proteins), whereas others were barely or not glycosylated. The glycosylated proglycinin was susceptible to endo-beta-N-acetylglucosamidase and N-glycanase cleavages. N-glycosylation did not cause inconveniences to processing of signal peptide, assembly into trimers and targeting into the vacuoles. Thermal and trypsin sensitivity analyses of the glycosylated proglycinin suggested that N-linked glycan prevents protein-protein interaction but does not stabilize the protein conformation. The reason why glycinin does not require N-glycosylation for its maturation is discussed.

Crystal structure of soybean proglycinin A1aB1b homotrimer.

Soybean glycinin is a member of the 11 S globulin family. The crystal structure of proglycinin was determined by X-ray crystallography at 2.8 A resolution with an R-factor of 0.199 and a free R-factor of 0.250. A trimer molecule was found in an asymmetric unit of crystals. The trimer model contains three A1aB1b subunits and comprises 1128 amino acid residues and 34 water molecules. The constituent protomers of the homo-trimeric protein are arranged around a 3-fold symmetry axis with dimensions of 95 Ax95 Ax40 A. The protomer model is composed of five fragments which correspond roughly to conserved regions based on the sequence alignment of various 11 S globulins. The core of the protomer consists of two jelly-roll beta-barrels and two extended helix domains. This structure of proglycinin is similar to those of canavalin and phaseolin belonging to the 7 S globulin family, strongly supporting the hypothesis that both 7 S and 11 S globulins are derived from a common ancestor. The inter and intra-chain disulfide bonds conserved in the 11 S globulin family are clearly observed. It is found that the face with the inter-chain disulfide bond (IE face) contains more hydrophobic residues than that with the intra-chain disulfide bond. This suggests that a mature hexamer is formed by the interaction between the IE faces after processing.

Expression of cytoplasmic galectin-3 as a prognostic marker in tongue carcinoma.

Galectin-3 is a member of the beta-galactoside-binding mammalian lectin family with affinity to ABH group epitopes, cell surface and extracellular polylactosamine glycans. It has been shown to be involved in differentiation, morphogenesis, tumor progression, and metastasis. Here we questioned the possible involvement of galectin-3 in the neoplastic progression of the tongue epithelium and evaluated its prognostic value in tongue cancer patients. Galectin-3 expression was analyzed by the immunohistochemical method in 77 tongue specimens (54 squamous cell carcinomas and 23 specimens of distinct normal mucosa). Levels of nuclear expression of galectin-3 markedly decreased during the progression from normal to cancerous states (P < 0.0001), while cytoplasmic expression increased (P < 0.0001). Enhanced expression of galectin-3 in the cytoplasm was associated with a reduced disease-free survival of tongue cancer patients. Multivariate analysis identified enhanced expression of cytoplasmic galectin-3 as an independent predictor of disease recurrence (P = 0.0120). These results suggest that the observed translocation of galectin-3 from the nucleus to the cytoplasm during neoplastic progression may serve as a prognostic factor for tongue cancer patients.

Comparative study of two Japanese rhinoconjunctivitis quality-of-life questionnaires.

CONCLUSION: Two questionnaires were used to assess quality of life (QOL) in allergic rhinitis: the Japanese translation of the Rhino-conjunctivitis Quality of Life Questionnaire (RQLQJ) and an original Japanese QOL questionnaire (JRQLQ). Either questionnaire may be used to assess QOL depending on differences in target domains. OBJECTIVES: Although pollinosis is a common disease which has a major impact on patient QOL, no internationally standardized questionnaire has been available in Japan until now. The aim of this study was to compare two currently available QOL questionnaires for allergic rhinitis in Japan-the RQLQJ and JRQLQ-in terms of their appropriateness for clinical use and their psychometric properties. MATERIAL AND METHODS: A multicenter, inter-group, cross-sectional study was conducted in 187 adult symptomatic patients with Japanese cedar pollinosis in 2003. Patient scores on the two questionnaires were compared in terms of both overall and comparable domains. We also examined the acceptability, construct and reliability of both questionnaires. RESULTS: The questionnaires were highly correlated in terms of both overall and comparable domain scores. In addition, both questionnaires had equal and satisfactory psychometric validity, demonstrating that they are both useful tools for assessing QOL in rhinitis. However, when compared with each other, the JRQLQ focuses mainly on activities of daily life and is simpler, while the RQLQJ focuses mainly on rhinitis-related health and is more responsive.

Expression of telomerase reverse transcriptase and telomere elongation during sexual maturation in Paramecium caudatum.

Paramecium caudatum has a sexually immature period that lasts for about 60 fissions. To examine the possibility that telomere length is one of the determining factors of the duration of immaturity, we cloned the telomerase reverse transcriptase (TERT) gene from P. caudatum, and analyzed its expression levels at mRNA, telomerase activity, and telomere length during the course of clonal division. Paramecium TERT (Pc_TERT) cDNA encodes a basic protein of 107 kDa that harbors conserved RT motifs, T motif, CP motif, and N motif. Pc_TERT mRNA is expressed at very low levels only detectable by RT-PCR, but constitutively, during immature and mature periods, exhibiting abundant telomerase activity. No clear phase shift in Pc_TERT expression, telomerase activity, or telomere length was observed at the point of maturation in P. caudatum. Instead, the telomere elongates successively as cells divide in P. caudatum, although a close species, P. tetraurelia, was reported to keep the length constant. We discuss possible mechanisms for the expression of sexual activity associated with telomere length in P. caudatum.

The 1,10-phenanthroline micelles-copper(I) complex catalyzes protein degradation.

The 1,10-phenanthroline micelles-copper(I) coordinated complex, not the mono-dispersed bis 1,10-phenanthroline-copper(I) one, rapidly degraded proteins in the presence of beta-mercaptoethanol in the neutral-to-mild acidic region under aerobic conditions. The degradation products derived from alpha-casein were peptides and amino acids.

Lack of correlation between timing of surgery in relation to the menstrual cycle and prognosis of premenopausal patients with early breast cancer.

In a retrospective cohort of a randomised study of adjuvant endocrine, chemotherapy and chemoendocrine therapy, we investigated the correlation between timing of mastectomy and relapse-free survival (RFS) and overall survival (OS) in 721 premenopausal patients with early breast cancer. The median follow-up was 10.1 years (range: 6.1-19.1 years). We grouped the patients by three kinds of classification according to Badwe, Senie, and Hrushesky. The logrank test after the Kaplan-Meier curves showed that there were no significant differences between the categorised menstrual phase in cycle and RFS or OS, except for RFS using Badwe's classification, where the patients whose timing of operation was in the follicular phase had a better RFS compared with those in the luteal phase. These were confirmed by the Cox proportional hazard model. These results suggest that a positive result might be a chance finding, dependent upon the cut-off levels in the menstrual cycle.

Prognostic impact of second cancer on the survival of early breast cancer patients.

Although numerous reports have been published on the incidence of second cancers after adjuvant therapy for early breast cancer, there have been few studies on the effect of the development of second cancer on overall survival (OS) of the patients. 1857 female patients younger than 80 years of age with operable breast cancer with UICC Stages I, II, and III who entered 3 trials of adjuvant therapy were studied for the detection of recurrence and second cancer. The median follow-up period for surviving patients was 12 years (range, 5-25 years). 384 of recurrence and 119 of second cancers occurred. 465 deaths were recorded, the causes of which were designated to be due to recurrence of breast cancer in 326 patients, second cancers in 57, and due to other causes in 82. Many background factors that were significantly related to recurrence did not influence the incidence of and death by second cancers after mastectomy: age and menopausal status alone were related. The difference in the Kaplan-Meier curves between the event (recurrence and second cancers)-free and relapse-free survivals indicates the incidence of second cancers is a significant event in post-operation course of early breast cancer patients (P=0.0001). The survival curve of patients who were free from recurrence- and second cancer-death is shown to be significantly lower than that of those free from breast cancer-specific death (P=0.0355), suggesting that death by second cancers is significantly influential to the overall survival. According to the Cox regression model using a recurrence or second cancers as time-dependent variables, the diagnosis of a recurrence or second cancer is shown to be highly significant to OS of the patients (hazard ratio: 49.3, and 6.3, respectively). Second cancers are shown to be not statistically significantly influential to the breast cancer specific survival (P=0.0637), nor a recurrence to second cancer specific survival (P=0.2285). In spite of heterogeneous distribution of sites of second primary cancers and their different natural histories, the incidence of second cancer has a significant effect on the postoperative survival of early breast cancer patients. In the survival analysis of early breast cancer patients, we have to estimate the contribution of second cancers to properly evaluate the effect of treatment.

Galectin-3 maintains the transformed phenotype of thyroid papillary carcinoma cells.

Galectin-3, a beta-galactoside-binding protein, is highly expressed in thyroid papillary carcinomas, while functional relevance of galectin-3 overexpression to the malignant phenotype remains elusive. In the present study we transfected galectin-3 antisense cDNA into the human thyroid papillary carcinoma cell line NPA which expresses an innately high level of galectin-3, and examined the effect of antisense inhibition of galectin-3 expression on the transformed phenotype. There was no difference in anchorage-dependent growth between the antisense clones and either the control or parental clones. In contrast, anchorage-independent growth and saturation density of the antisense clones were significantly suppressed compared to those of either the control or parental clones. These results demonstrate that overexpression of galectin-3 in thyroid papillary carcinoma cells is necessary for the maintenance of transformed phenotype, and suggest galectin-3 as a potential target for therapeutic interventions in the future.

Polyenthesitis.

We described the clinical symptoms and signs of 11 cases of polyenthesitis, confirmed by the presence of inflammation in biopsies and increased activity in scintigraphy. All cases are negative for HLA-B27, and there is no presence of systemic inflammatory reaction, radiographic sacroiliitis, or chronic arthritis. The enthesitis is not ossifying and follows a rather benign clinical course. In one case, herpes virus-like particles were detected in the fibroblast debris of the tissue removed from Achilles tendon insertion. This fact suggests that polyenthesitis may be a clinical manifestation due to a reaction to an infectious agent.

Selective portal branch occlusion by balloon catheter during liver resection.

An intraoperative ultrasonographically guided introduction of a balloon catheter into labor or smaller branches of the portal vein within the liver parenchyma made it possible to temporarily occlude them and perform regional staining during resection for tumors. The technique minimized blood loss without hilar dissection for vascular control, and the presence of the catheter facilitated intraparenchymal dissection of the portal stalk to the part of the liver to be resected.

Carcinoma of the bifurcation of the hepatic ducts.

Thirty-one patients with carcinoma of the bifurcation of the hepatic ducts were treated during a period of ten years. Sixteen of the 31 patients underwent resection. Left, right, and extended right lobectomy with the resection of the bilateral hepatic ducts, common hepatic and bile ducts were performed on 11, 1, and 3 patients, respectively. In a cirrhotic patient, resection of the left medial segment and bile ducts was carried out. The biliary tract was reconstructed using hepaticojejunostomy or hepatojejunostomy. Four of the 16 patients are still alive for six months, 1 1/2 years, one year and 11 months, and two years and ten months, respectively. The remaining 12 patients died of recurrence at periods ranging from six months to three years and eight months. The treatment is now focused on increasing long-term survival.

Designing of an orally active complement C3a agonist peptide with anti-analgesic and anti-amnesic activity.

Complement C3a is an anti-opioid peptide, having anti-analgesic and anti-amnesic effects after intracerebroventricular administration. However, the peptide is inactive after oral administration. Orally active C3a agonist peptide was designed based on the structure of oryzatensin, a C3a agonist peptide derived from rice albumin. Tyr-Pro-Leu-Pro-Arg, a pentapeptide at the carboxyl terminus of oryzatensin is the minimally essential structure for exerting C3a activity. Due to the affinity for mu-opioid receptor, both oryzatensin and Tyr-Pro-Leu-Pro-Arg showed analgesia after intracerebroventricular administration in mice which was blocked by the opioid antagonist naloxone. Tyr-Pro-Leu-Pro-Arg lost opioid activity by substitution the amino terminus tyrosine with other hydrophobic residues. Among the newly designed peptides, Trp-Pro-Leu-Pro-Arg was found to possess the strongest C3a activity. The peptide antagonized morphine-induced analgesia at 300 mg/kg after oral administration and also improved scopolamine- and ischemia-induced amnesia in a step-through passive avoidance test.