Search

Aims: To develop a rapid and sensitive method for detecting Brucella spp. Methods and Results: Two sets of six Brucella-specific primers for loop-mediated isothermal amplification (LAMP) were designed from the sequence of the Brucella abortus BCSP31 gene. The specificity and sensitivity were examined for six Brucella species (22 strains) and 18 non- Brucella species (28 strains). The LAMP assay was specific to Brucella spp. in 35 min at 63°C and sensitive (detected 10 fg of genomic DNA). The assay was also applied for the detection of Brucella DNA in contaminated milk and infected mouse organs. Conclusions: We developed a sensitive and specific LAMP assay for Brucella spp., with the test appearing to be useful for the detection of the pathogen from clinical and food samples. Significance and Impact of the Study: This is the first report of the development of LAMP for the detection of Brucella spp. As the LAMP assay can be performed at a constant temperature and its reactivity is directly observed with the naked eye without electrophoresis, our assay should be useful for the diagnosis of brucellosis as well as the detection of the bacteria in environmental or food samples. [ABSTRACT FROM AUTHOR]

Abstract: Background: Hyaluronan (HA) is an important constituent of extracellular matrix (ECM) in the skin, and HA degradation mediated by HYBID (KIAA1199) is suggested to be implicated in facial skin wrinkling in Japanese women. Ethnic difference in skin wrinkle formation is known between Caucasian and Japanese women, but no information is available for the relations of HA and HYBID expression levels with skin wrinkling in Caucasian women. Methods: The skin surface roughness at the eye corner of the Caucasian female subjects was measured, and the skin specimens biopsied from the same areas were subjected to microarray gene analysis, HA staining, and immunohistochemistry for HYBID. Results: Among the ECM genes and those related to ECM metabolism, only HYBID expression levels positively correlated with the skin roughness parameters. When the skin sample groups with high expression of HYBID or low expression of HYBID were compared, the HA staining intensity and the ratio of HYBID‐immunoreactive cells to total cells in the superficial dermis were significantly reduced and increased in the high‐HYBID‐expression group compared with the low‐HYBID‐expression group, respectively. Conclusion: Our data suggest that like Japanese women, HYBID‐mediated reduction of HA in the superficial dermis is involved in the formation of wrinkles in Caucasian women. [ABSTRACT FROM AUTHOR]

Summary: Background: Hyaluronan (HA) metabolism in skin fibroblasts is mediated by HYBID (hyaluronan binding protein involved in hyaluronan depolymerization, alias CEMIP and KIAA1199) and the HA synthases HAS1 and HAS2. However, photoageing-dependent changes in HA and their molecular mechanisms, and the relationship between HA metabolism and clinical symptoms in photoaged skin remain elusive. Objectives: We examined the amount, size and tissue distribution of HA and expression levels of HYBID, HAS1 and HAS2 in photoaged skin, and analysed their relationship with the degree of photoageing. Methods: Photoageing-dependent changes of HA were investigated by studying skin biopsies isolated from photoprotected and photoexposed areas of the same donors, and the relationships between HA and photoageing symptoms such as skin wrinkling and sagging were examined. Results: Skin biopsy specimens showed that the amount and size of HA are decreased in photoexposed skin compared with photoprotected skin, and this was accompanied by increased expression of HYBID and decreased expression of HAS1 and HAS2. Histologically, HA staining in the papillary dermis was decreased in photoexposed skin, showing reverse correlation with HYBID expression. HYBID expression in the photoexposed skin directly correlated with skin roughness and sagging parameters, and the reduced HA staining in the papillary dermis in the photoexposed skin positively correlated with these symptoms. Conclusions: These data demonstrate that imbalance between HYBID-mediated HA degradation and HAS-mediated HA synthesis may contribute to enhanced HA catabolism in photoaged skin, and suggest that HYBID-mediated HA reduction in the papillary dermis is related to skin wrinkling and sagging of photoaged skin. What's already known about this topic? Hyaluronan (HA) is an important determinant of skin homeostasis. HYBID (hyaluronan binding protein involved in hyaluronan depolymerization, alias CEMIP and KIAA1199) and HA synthases (HAS1 and HAS2) play a key role in HA metabolism in skin fibroblasts.Major alterations in photoaged skin have been demonstrated in the form of accumulation of abnormal elastin and severe loss of collagens.Photoageing-dependent changes of HA and their molecular mechanisms, and the relationship between HA metabolism and clinical symptoms in photoaged skin remain elusive. What does this study add? HA homeostasis becomes unbalanced in photoaged skin through increased HYBID-mediated HA degradation compared with HAS1- and HAS2-mediated HA synthesis.Loss of HA in the papillary dermis and HYBID expression levels directly correlated with the degree of skin wrinkling and sagging of photoaged skin.These findings suggest that the HYBID-mediated decrease of HA in the papillary dermis seems to contribute to skin wrinkling and sagging in photoaged skin. What is the translational message? Strategies to downregulate HYBID expression or inhibit HYBID-mediated HA degradation in the papillary dermis could provide new remedies to prevent or improve photoageing-dependent skin damage. Respond to this article [ABSTRACT FROM AUTHOR]