[Nationwide Survey of Medical Radiation Exposure on Cardiovascular Examinations in Japan].

PURPOSE: It is very important to manage the radiation dose of cardiovascular interventional (CVI) procedures. Overseas, the diagnostic reference levels for cardiac interventional procedures were established with the air kerma at the patient entrance reference point (Ka,r) and the air kerma-area product (PKA). Although the Japan DRLs 2015 was established by the Japan Network for Research and Information on Medical Exposure (J-RIME), the Japan DRL for CVIs were established by fluoroscopic dose rates of 20 mGy/min at the patient entrance reference point with 20 cm thickness polymethyl methacrylate (PMMA) phantom. In the present our study, we performed a questionnaire survey of indicated values of angiographic parameters in CVI procedures. METHODS: A nationwide questionnaire was sent by post to 765 facilities. Question focused on angiographic technology, exposure parameters and radiation doses as the displayed dosimetric parameters on the angiographic machine. RESULTS: The recovery rate was 22.8% at 175 out of 765 facilities. In total 1728 cases of the coronary angiography (CAG), 1703 cases of the percutaneous coronary intervention (PCI), 962 cases of the radiofrequency catheter ablation (RFCA) and 377 cases of pediatric CVI. The 75th percentile value of Ka,r, PKA, fluoroscopy time (FT) and number of cine images (CI) for CAG, PCI, RFCA and pediatric CVI were 702, 2042, 644, and 159 mGy, respectively, 59.3, 152, 81.3, and 14.9 Gy・cm2, respectively, 10.2, 35.6, 61.1, and 35.6 min, respectively and 1503, 2672, 722, and 2378 images, respectively. Our investigation showed that the angiographic parameters were different in several CVI procedures. CONCLUSIONS: The displayed dosimetric parameters on the angiographic machine in CVI procedures showed different values. We should classify the dosimetric parameters for each procedure.

Epidemiological and pathological study of feline morbillivirus infection in domestic cats in Japan.

BACKGROUND: Feline morbillivirus (FmoPV) is a novel paramyxovirus found to infect domestic cats. FmoPV has been isolated in several countries in Asia and Europe and is considered to have genetic diversity. Also, it is suspected to be associated with feline renal diseases including tubulointerstitial nephritis (TIN), which affects domestic cats with a high incidence rate. RESULTS: To clarify the state of FmoPV infection among domestic cats in Japan, an epidemiological survey was conducted. Twenty-one out of 100 cats were found to have serum antibodies (Ab) against FmoPV-N protein by indirect immunofluorescence assay (IF) using FmoPV-N protein-expressing HeLa cells. Twenty-two of the cats were positive for FmoPV RNA in the urine and/or renal tissues. In total, 29 cats were positive for Ab and/or viral RNA. These FmoPV-infected cats were classified into three different phases of infection: RNA+/Ab + (14 cats), RNA+/Ab- (8 cats) and RNA-/Ab + (7 cats). In immunohistochemistry (IHC), 19 out of 29 cats were positive for FmoPV-N protein in kidney tissues; however, the FmoPV-N protein was located in the inflammatory lesions with severe grade in only four out of the 19 cats. Since 15 out of 29 infected cats were positive for viral RNA and Ab, approximately half of the infected cats were persistently infected with FmoPV. CONCLUSIONS: A statistically significant difference was observed between infection of FmoPV and the presence of inflammatory changes in renal lesions, indicating a relationship between FmoPV infection and feline renal diseases. However, we could not obtain histopathological evidence of a relationship between FmoPV infection and TIN.

ZIP2 protein, a zinc transporter, is associated with keratinocyte differentiation.

Zinc is essential for the proper functioning of various enzymes and transcription factors, and its homeostasis is rigorously controlled by zinc transporters (SLC39/ZIP, importers; SLC30/ZnT, exporters). Skin disease is commonly caused by a zinc deficiency. Dietary and inherited zinc deficiencies are known to cause alopecia and the development of vesicular or pustular dermatitis. A previous study demonstrated that zinc played crucial roles in the survival of keratinocytes and their unique functions. High levels of zinc have been detected in the epidermis. Epidermal layers are considered to use a mechanism that preferentially takes in zinc, which is involved with the unique functions of keratinocytes. However, few studies have investigated the ZIP (Zrt- and Irt-like protein) proteins specifically expressed in keratinocytes and their functions. We explored the ZIP proteins specifically expressed in the epidermis and analyzed their functions. Gene expression analysis showed that the expression of ZIP2 was consistently higher in the epidermis than in the dermis. Immunohistochemistry analysis confirmed the expression of ZIP2 in differentiating keratinocytes. The expression of ZIP2 was found to be up-regulated by the differentiation induction of cultured keratinocytes. Intracellular zinc levels were decreased in keratinocytes when ZIP2 was knocked down by siRNA, and this subsequently inhibited the differentiation of keratinocytes. Moreover, we demonstrated that ZIP2 knockdown inhibited the normal formation of a three-dimensional cultured epidermis. Taken together, the results of this study suggest that ZIP2, a zinc transporter expressed specifically in the epidermis, and zinc taken up by ZIP2 are necessary for the differentiation of keratinocytes.

Senescent dermal fibroblasts enhance stem cell migration through CCL2/CCR2 axis.

During aging, increases in the number of senescent cells are seen in various tissues. On the other hand, stem cells play crucial roles in tissue repair and homeostasis. Therefore, it is likely that stem cells give rise to new cells that replace senescent cells. However, how stem cells contribute to homeostasis in the dermis has not been elucidated. Here, we investigated the effects of factors secreted from senescent fibroblasts on stem cells. We found that senescent human dermal fibroblast (HDF) conditioned medium (CM) significantly enhanced stem cell migration compared with young HDF CM. The senescent HDF CM strongly secreted chemokine ligand 2 (CCL2). Furthermore, CCL2 was found to enhance stem cell migration, and the inhibition of CCR2, a receptor for CCL2, reduced stem cell migration. These results suggest that senescent fibroblasts recruit stem cells by secreting various factors and that the CCL2/CCR2 axis is one of the mechanisms underlying this phenomenon.

Accelerated differentiation of melanocyte stem cells contributes to the formation of hyperpigmented maculae.

It has been reported that the abnormal regulation of melanocyte stem cells (McSCs) causes hair greying; however, little is known about the role of McSCs in skin hyperpigmentation such as solar lentigines (SLs). To investigate the involvement of McSCs in SLs, the canonical Wnt signalling pathway that triggers the differentiation of McSCs was analysed in UVB-induced delayed hyperpigmented maculae in mice and human SL lesions. After inducing hyperpigmented maculae on dorsal skin of F1 mice of HR-1× HR/De, which was formed long after repeated UVB irradiation, the epidermal Wnt1 expression and the number of nuclear ß-catenin-positive McSCs were increased as compared to non-irradiated control mice. Furthermore, the expression of dopachrome tautomerase (Dct), a downstream target of ß-catenin, was significantly upregulated in McSCs of UVB-irradiated mice. The Wnt1 expression and the number of nuclear ß-catenin-positive McSCs were also higher in human SL lesions than in normal skin. Recombinant Wnt1 protein induced melanocyte-related genes including Dct in early-passage normal human melanocytes (NHEMs), an in vitro McSC model. These results demonstrate that the canonical Wnt signalling pathway is activated in SL lesions and strongly suggest that the accelerated differentiation of McSCs is involved in SL pathogenesis.

Bleomycin inhibits adipogenesis and accelerates fibrosis in the subcutaneous adipose layer through TGF-ß1.

Systemic sclerosis [scleroderma (SSc)]-associated skin fibrosis is characterized by increased fibrosis in the dermis and a reduction in the thickness of the subcutaneous adipose tissue layer. Although many studies have examined fibrosis in SSc, only a few studies have focused on the associated reduction in the thickness of the subcutaneous adipose tissue layer. In this study, we investigated the effects of SSc-induced fibrosis on adipose tissue. We found that bleomycin suppresses adipogenesis in adipose-derived stem cells (ASCs) and stimulates ASCs to express transforming growth factor ß1 (TGF-ß1), which suppresses adipogenesis and promotes fibrosis. Furthermore, we found that adipocyte-conditioned medium suppressed collagen synthesis by fibroblasts in fibrosis-like conditions. We concluded that in the skin affected by bleomycin-induced fibrosis, increased TGF-ß1 expression suppresses adipogenesis and promotes adipocyte fibrosis. It was also suggested that adipocytes have an inhibitory effect on the progression of fibrosis.

Analysis of the effects of hydroquinone and arbutin on the differentiation of melanocytes.

Hydroquinone (HQ) is a chemical compound that inhibits the functions of melanocytes and has long been known for its skin-whitening effect. According to previous studies, the Tyrosinase (Tyr) activity inhibitory effect and melanocyte-specific cell toxicity are known depigmenting mechanisms; however, details of the underlying mechanisms are unknown. Arbutin (Arb) is also known for its Tyr activity inhibitory effect and is commonly used as a skin-whitening agent. However, the detailed depigmenting mechanism of Arb is also not yet fully understood. Few studies have attempted to elucidate the effects of HQ and Arb on undifferentiated melanocytes. In this study, we examined the effects of HQ and Arb throughout each stage of differentiation of melanocytes using a mouse embryonic stem cell (ESC) culture system to induce melanocytes. The results showed that HQ in particular downregulated the early stage of differentiation, in which neural crest cells were generated, and the late stage of differentiation, in which melanogenesis became active. On the other hand, Arb had no effect on the differentiation of melanocytes, and only suppressed melanogenesis by specifically suppressing elevations in Tyr expression in the late stage of differentiation.

Increased hydrophobicity in Malassezia species correlates with increased proinflammatory cytokine expression in human keratinocytes.

Malassezia cells stimulate cytokine production by keratinocytes, although this ability differs among Malassezia species for unknown reasons. The aim of this study was to clarify the factors determining the ability to induce cytokine production by human keratinocytes in response to Malassezia species. M. furfur NBRC 0656, M. sympodialis CBS 7222, M. dermatis JCM 11348, M. globosa CBS 7966, M. restricta CBS 7877, and three strains each of M. globosa, M. restricta, M. dermatis, M. sympodialis, and M. furfur maintained under various culture conditions were used. Normal human epidermal keratinocytes (NHEKs) (1 × 10(5) cells) and the Malassezia species (1 × 10(6) cells) were co-cultured, and IL-1α, IL-6, and IL-8 mRNA levels were determined. Moreover, the hydrophobicity and ß-1,3-glucan expression at the surface of Malassezia cells were analyzed. The ability of Malassezia cells to trigger the mRNA expression of proinflammatory cytokines in NHEKs differed with the species and conditions and was dependent upon the hydrophobicity of Malassezia cells not ß-1,3-glucan expression.

Melanocyte stem cells express receptors for canonical Wnt-signaling pathway on their surface.

It has been reported that melanocytes play important roles in skin and hair pigmentation and are differentiated from melanocyte stem cells (MSCs) residing in the bulge area of hair follicles. Recently, interest has been growing in MSCs because regulation of the upstream of differentiated melanocytes is essential for the determination of skin and hair pigmentation; however, their precise characteristics remain to be elucidated. The aim of this study is to explore cell-surface markers expressed on MSCs in order to understand their characteristics. To explore genes specifically expressed in the bulge region, we classified a hair follicle into four areas, hair bulb, hair bulb to bulge (lower bulge), bulge, and epidermis to bulge (upper bulge), and collected these areas from back skin sections of C57BL/6 mice by laser microdissection. Real-time RT-PCR performed on these areas revealed that Frizzled (Fzd)-4, Fzd7, low density lipoprotein receptor-related protein 5 (Lrp5), and Lrp6, receptors for Wnt molecules, were expressed higher in the bulge area than other areas. Furthermore, FACS analysis showed that populations of Fzd4(+) cells and Fzd7(+) cells were different from those of Kit(+) cells (precursor of melanocytes: melanoblasts). Fzd4(+) and Fzd7(+) cells isolated by FACS required a longer culture period to differentiate into mature melanocytes than Kit(+) cells. Up-regulation of mRNA expressions of melanocyte markers (dopa chrometautomerase: Dct, tyrosinase: Tyr, tyrosinase-related protein 1: Tyrp1) was observed in Fzd4(+) and Fzd7(+) cells following Kit(+) cells during differentiation. These results suggested that Fzd4(+) and Fzd7(+) cells were more immature than melanoblasts, therefore raising the possibility that Fzd4(+) and Fzd7(+) cells are MSCs.

Cutaneous Malassezia microbiota in atopic dermatitis patients differ by gender and body part.

BACKGROUND: Malassezia is a particularly important factor in the occurrence of atopic dermatitis (AD). AIM: The aim of this study was to quantitatively clarify the Malassezia species isolated from AD patients by gender, body part and analytical method in detail. METHODS: The subjects were 20 AD males and 47 AD females. Samples were collected from lesion and nonlesion areas on the face and upper trunk of AD patients. Malassezia DNA was analyzed using a real-time PCR system. RESULTS: The cutaneous Malassezia microbiota in AD patients differed by gender, body part and analytical method. CONCLUSIONS: The present results indicate the possibility that the influence of Malassezia antigens is different according to gender and body part.

Effects of different food colorants and polishing techniques on color stability of provisional prosthetic materials.

The main objective was to investigate the effects of different polishing techniques on the color stability of provisional prosthetic materials upon exposure to different staining agents by mimicking the oral environment in vitro. Fifty-six cylindrical specimens were prepared for each type of material: bis-acryl and light-polymerized composite resins, and methyl methacrylate- and ethyl methacrylatebased resins. The specimens were polished using seven different polishing techniques and then immersed in four different staining agents. Color was measured with a spectrophotometer before and after immersion, and color changes (DeltaE) were calculated. The effects of the type of provisional material, polishing procedure, staining agent, and their interactions on color stability were significant (p<0.05). Amongst these factors, the staining agent exerted the strongest effect on color stability. Amongst the provisional materials tested, methacrylate-based resins exhibited the highest color stability irrespective of polishing technique and staining agent.

Human leptospirosis cases and the prevalence of rats harbouring Leptospira interrogans in urban areas of Tokyo, Japan.

Thirteen patients with leptospirosis were identified, as confirmed by laboratory analysis during the last 5 years in our laboratory, who came from urban areas of Tokyo, Japan. All of the patients came into contact with rats before the onset of illness. Seventeen per cent of Norway rats captured in the inner cities of Tokyo carried leptospires in their kidneys. Most of these rat isolates were Leptospira interrogans serovar Copenhageni/Icterohaemorrhagiae. Antibodies against these serovars and their DNA were detected in the patients. This suggests that rats are important reservoirs of leptospirosis, and that rat-borne leptospires occur in urban areas of Tokyo.

Malassezia folliculitis is caused by cutaneous resident Malassezia species.

Malassezia folliculitis [MF] is caused by the invasion of hair follicles by large numbers of Malassezia cells, but it remains unclear which Malassezia species are involved in the disease. To clarify this situation, Malassezia species isolated from lesions of MF patients were analyzed by both culture and non-culture methods. In addition, Malassezia species recovered from the non-lesion areas of the skin of MF patients and skin samples of healthy subjects were included in this study. The test population consisted of 32 MF patients and 40 healthy individuals. The lesions were obtained using a comedone extractor, while swabs were employed to obtain skin samples from non-lesion areas of the patients and healthy subjects. Malassezia DNA was analyzed using a real-time PCR technique. The detection limit of the culture method was 5 CFU/cm(2) as opposes 50 cells/cm(2) with non-culture procedures. The predominant species recovered from MF lesions were M. globosa and M. sympodialis by culture method analysis, and M. restricta, M. globosa, and M. sympodialis with non-culture methods. These results were in agreement with those found with samples from non-lesion skin areas of MF patients and healthy subjects. This study clarified that MF is caused by Malassezia species that are part of the cutaneous microflora and not by exogenous species.

Fermentable metabolite of Zymomonas mobilis controls collagen reduction in photoaging skin by improving TGF-beta/Smad signaling suppression.

Solar ultraviolet (UV) irradiation causes damages on human skin and premature skin aging (photoaging). UV-induced reduction of type I collagen in dermis is widely considered primarily induction of wrinkled appearance of photoaging skin. Type I procollagen synthesis is reduced under UV irradiation by blocking transforming growth factor-beta (TGF-beta)/Smad signaling; more specifically, it is down-regulation of TGF-beta type II receptor (T beta RII). Therefore, preventing UV-induced loss of T beta RII results decreased type I collagen reduction in photoaging skin. Zymomonas mobilis is an alcohol fermentable, gram-negative facultative anaerobic bacterium whose effect on skin tissue is scarcely studied. We investigated the protective effects of fermentable metabolite of Z. mobilis (FM of Z. mobilis) against reduction of type I procollagen synthesis of UV-induced down-regulation of T beta RII in human dermal fibroblasts FM of Z. mobilis was obtained from lyophilization of bacterium culture supernatant. The levels of T beta RII and type I procollagen mRNA in human dermal fibroblasts were measured by quantitative real-time RT-PCR, and T beta RII protein levels were assayed by western blotting. T beta RII, type I procollagen, and type I collagen proteins in human dermal fibroblasts or hairless mouse skin were detected by immunostaining. FM of Z. mobilis inhibited down regulation of T beta RII mRNA, and protein levels in UVB irradiated human dermal fibroblasts consequently recover reduced type I procollagen synthesis. These results indicate UVB irradiation inhibits type I procollagen synthesis by suppression of TGF-beta/Smad signaling pathway, and FM of Z. mobilis has inhibitory effect on UVB-induced reduction of type I procollagen synthesis. While short period UVB irradiation decreased both T beta RII and type I procollagen protein levels in hairless mouse skin, topical application of FM of Z. mobilis prevented this decrease. Wrinkle formation in hairless mouse skin surface was accelerated by continuous 5 month UVB irradiation along with a reduction of type I collagen in the dermis, but this change was prevented by topical application of FM of Z. mobilis. From this experimental data, it is suggested that FM of Z. mobilis is effective for suppression of wrinkle formation in photoaging skin by inhibition of type I procollagen synthesis reduction.

Conventional and early loading of two-implant supported mandibular overdentures. A systematic review.

OBJECTIVES: Implant loading time is considered to influence the treatment outcomes. Number of experimental studies have shown that implant loading up to 3 months can produce equally satisfactory results. However, research results in this area are not consistent. The purpose of the study was to investigate the influence of conventional and early loading of two-implant supported mandibular overdentures on treatment outcomes. METHODS: The articles from 1985 to 2007 in English related to the topic were identified. Totally 221 (214 online and 7 printed) primary articles were detected. Eight articles were selected for data extraction. Implant survival and success rates, periimplant parameters, prosthetic maintenance and patient satisfaction were considered. RESULTS: Implant success rate ranged from 83% to 100% in conventional loading group and from 71% to 100% in early loading group. During the first year in conventional group, the marginal bone loss ranged from 0.35 to 0.91 mm, during the second year--from zero to 0.2 mm, whereas in early loading group these intervals were 0.12-1 mm and 0-0.15 mm, respectively. Comparing averaged probing depth values at different time periods, it could be noticed that around conventionally loaded implants probing depth slightly decreased (from 1.62 mm to 1.56 mm), while around early loaded implants--increased from 1.7 mm to 1.82 mm. CONCLUSIONS: Considering implant success rates and peri-implant parameters early loading protocol produces equal outcomes as with conventional loading. More well designed studies are needed to further substantiate the early loading protocol.

Maxillary complete denture outcome with two-implant supported mandibular overdentures. A systematic review.

OBJECTIVES: Research data regarding maxillary complete denture outcome with two-implant supported mandibular overdentures are not consistent. Considering multiple publications on implant supported mandibular overdentures, it was decided to summarize currently present evidence on the maxillary complete dentures opposed by implant-supported mandibular overdentures, and analyze factors that could potentially influence the outcomes. METHODS: The articles from 1985 to 2007 related to the topic were identified in the online MEDLINE/Pubmed and other databases and manually. Primary articles were scanned, and irrelevant studies were excluded from the further review process. Potentially relevant titles and abstracts were provisionally included for consideration on the basis of full text articles. Full text articles were obtained from on-line and printed sources. The data from the studies were extracted and reviewed. RESULTS: The study has failed to identify any prospective satisfying inclusion/exclusion criteria RCT reporting on maxillary bone resorption. The number of maxillary complete denture relining incidences per patient was constantly increasing during the 10-year period. Maxillary complete denture remake incidences comprised 16-33 % of the number of patients followed during the 10-year period. Comparing patient satisfaction with upper dentures at the baseline and after two years, no decrease in satisfaction was noticed. CONCLUSIONS: There is no evidence that maxillary ridge resorption is accelerated with certain types of two-implant supported mandibular overdenture attachments. Most common complication for the maxilla - prosthetic maintenance. There is a risk of decreased patient satisfaction with bar-supported mandibular overdenture. Further studies are needed to provide evidence for the maxillary complete denture outcome with two-implant supported mandibular overdentures.

Metallic artifacts in MRI caused by dental alloys and magnetic keeper.

PURPOSE: To evaluate the artifacts generated by crownshaped dental alloys and a magnetic keeper quantitatively by analyzing digital MRI data. METHODS: One pre-fabricated magnetic keeper and four clinical dental alloys (gold-silver-palladium, casting gold alloy Type 3, cobalt-chromium, gold porcelain alloy) were selected. Twenty metal crowns and 5 magnetic keepers were analyzed. The samples were placed in an acrylic phantom (150 mmx150 mmx150 mm) filled with agar, and then placed in the MRI apparatus. Various image slices were selected from the center (0 mm) to 70 mm at steps of 5 mm. The distribution of the signal intensity in the region of interest was calculated using ImageJ software and the mean coefficient of variation of each specimen was obtained. Statistical analysis was performed by Dunnett's test (p<0.05). RESULTS: Compared to the resin control, cobalt-chromium showed significantly greater signal intensity up to 40 mm in coronal T2-WI images and up to 70 mm in axial T1-WI images for the magnetic keeper. The signal intensities of gold-silver-palladium and casting gold alloy Type 3 were not significantly different from that of the control. The signal intensity of gold porcelain alloy was significantly different from that of the control at 0 mm and 5 mm in coronal T1,T2-WI and in sagittal T1-WI at 0 mm. CONCLUSIONS: The artifacts generated by the magnetic keeper and the cobalt-chromium crown when they are used in a second molar can disturb the MR images of the temporomandibular joint.

Age-related decrease in CD271(+) cells in human skin.

According to recent studies, stem cells are found in various tissues in our bodies. It has been reported that stem cells can reside in the skin tissues, including the epidermis, dermis, hair follicles and subcutaneous tissues. Homeostasis of the skin is maintained because these stem cells collaborate with each other to form new cells. We previously identified the CD271(p75NTR)(+) cell as a stem cell that was present in the epidermis, dermis and subcutaneous tissue, and further investigated the role of stem cells in wound healing and their association with skin disease. In this study, we investigated the localization of CD271(+) cells in human skin (epidermis and dermis) and its age-related changes in stem cells using CD271(+) cells. The study revealed that the number of CD271(+) cells in the epidermis and dermis decreased with aging. It is possible that such an age-related decrease in stem cells causes impaired regenerative ability and is associated with various skin diseases. If the relationship between stem cells and skin aging and diseases can be elucidated by investigations such as this study, it may lead to the development of novel anti-aging technologies and medical treatments for skin diseases in the future.

Advanced oxidation effect of ozonation combined with electrolysis.

The advanced oxidation effect of ozonation combined with electrolysis (electrolysis-ozonation) was discussed through the treatment of 4-chlorobenzoic acid (4-CBA) as a hydroxyl radical probe. The mechanism of hydroxyl radical production by electrolysis-ozonation process was also estimated with a mathematical model. The experimental results revealed that the electrolysis-ozonation process had a synergistic effect on the degradation of 4-CBA. The advanced oxidation effect of electrolysis-ozonation was inferred from standard potentials of relevant electrochemical reactions and mathematical model analysis to be mainly attributed to ()O(3)(-) promotion of O(3) at the cathodes. An increase in electric current improved the degradation rate of 4-CBA. However, the pseudo-first order degradation rate constant reached a plateau at high electric current densities, as ()O(3)(-) promotion of O(3) at the cathodes was regulated by O(3) transport process from the bulk to the cathodes in the range exceeding an electric current density of 10 Am(-2). Accordingly, the balance of O(3) transport flux and electric current is important for the efficient operation of the electrolysis-ozonation reactor.