Solid-Phase-Supported Chemoenzymatic Synthesis and Analysis of Chondroitin Sulfate Proteoglycan Glycopeptides.

Proteoglycans (PGs), consisting of glycosaminoglycans (GAGs) linked with the core protein through a tetrasaccharide linkage region, play roles in many important biological events. The chemical synthesis of PG glycopeptides is extremely challenging. In this work, the enzymes required for synthesis of chondroitin sulfate (CS) PG (CSPG) have been expressed and the suitable sequence of enzymatic reactions has been established. To expedite CSPG synthesis, the peptide acceptor was immobilized on solid phase and the glycan units were directly installed enzymatically onto the peptide. Subsequent enzymatic chain elongation and sulfation led to the successful synthesis of CSPG glycopeptides. The CS dodecasaccharide glycopeptide was the longest homogeneous CS glycopeptide synthesized to date. The enzymatic synthesis was much more efficient than the chemical synthesis of the corresponding CS glycopeptides, which could reduce the total number of synthetic steps by 80 %. The structures of the CS glycopeptides were confirmed by mass spectrometry analysis and NMR studies. In addition, the interactions between the CS glycopeptides and cathepsin G were studied. The sulfation of glycan chain was found to be important for binding with cathepsin G. This efficient chemoenzymatic strategy opens new avenues to investigate the structures and functions of PGs.

Transformation from Marginalization to Society's Agent: A Narrative Inquiry on Experiences of Art Activity among People with Mental Illness.

Using in-depth interviews, this study identified the experiences of five people with mental illness who participated in art activities. Findings indicated that the life journey that led to the participants' art activities and the impact of these activities in their lives were different and unique. The voices of the individuals with mental illness who participated in this study provided deeper understanding of how art activities have changed their lives. In addition, it was confirmed that art activities in which people with mental illness participate, bring positive inner changes and have the potential to improve their interpersonal relationships and social interactions.

"Moving forward despite adversity": The lives of Korean older adults with schizophrenia.

INTRODUCTION: Older adults with schizophrenia experience aging-related challenges and chronic psychiatric difficulties. However, their uniqueness is understudied. Aim This study explored three life stories to illuminate schizophrenia's trajectory. METHOD: A narrative inquiry method was used to elicit rich narratives in chronic patients' lives. Two men and one woman suffering from chronic schizophrenia for more than 20 years were engaged in conversation three times. RESULTS: The analysis revealed one main theme expressing their fundamental perspective on their lives: "moving forward despite adversity." Three subthemes emerged: living with the mental disorder, giving and receiving support, and adaptation to old age. They reflected on their interpersonal relationships, failures, and successes in the context of the challenges of schizophrenia. They indicated adapting to schizophrenia, exchanging social support with loved ones, and meeting aging-related challenges with confidence. They were hopeful about brighter personal futures than their pasts. DISCUSSION: Family and community supports were very important; self-acceptance positively influenced their successful community living. IMPLICATIONS FOR PRACTICE: For effective long-term treatment of older adults with schizophrenia, mental health policies should focus on meeting the social needs of patients and families and reorient programs away from the medical model limited to symptom reduction.

Whitening effect of novel peptide mixture by regulating melanosome biogenesis, transfer and degradation.

Peptides are short chain of amino acids linked by peptide bonds. They are widely used as effective and biocompatible active ingredients in cosmetic industry. In this study, we developed novel peptide mixture and identified its anti-pigmentation effect on melanocytes and keratinocytes. Our results revealed that peptide mixture inhibited melanosome biogenesis through the regulation of microphthalmia-associated transcription factor, a key factor of melanogenesis in melanocytes. And we observed that peptide mixture inhibited melanosome uptake through the reduction of protease-activated receptor 2, a phagocytosis-related receptor in keratinocytes. Furthermore, peptide mixture activated autophagy system resulting in degradation of transferred melanosomes in keratinocytes. The anti-pigmentation effect of multi-targeting peptide mixture was assessed in a human skin equivalent model (MelanoDerm). Melanin contents in epidermal layer were significantly decreased by topical treatment of peptide mixture, suggesting that it can be applied as a novel cosmetics material having a whitening function.

Leucine-rich glioma inactivated 3 is a melanogenic cytokine in human skin.

Recently, we demonstrated that leucine-rich glioma inactivated 3 (LGI3) is expressed in human skin. However, the effects of LGI3 on melanocytes remain unknown. The present study demonstrated that LGI3 can serve to stimulate melanogenesis without affecting cell viability. To determine the effects of LGI3 on melanin synthesis, normal human melanocytes and Mel-Ab cells were treated with recombinant LGI3 and melanin content was measured. Our results showed that LGI3 promoted melanin synthesis in both cell types. Moreover, upregulation of microphthalmia-associated transcription factor (MITF) and tyrosinase was observed at both the mRNA and protein levels via RT-PCR and Western blotting, respectively. Furthermore, immunohistochemical staining showed that the expression of LGI3 increased in the basal layer of melasma skin samples, whereas it decreased slightly in vitiligo samples. These results suggest that LGI3 may play a role as a melanogenic cytokine in human skin.

Controllable synthesis of thiol-capped CdTe nanoparticles for optical sensing of triethylenetetramine dihydrochloride.

Highly luminescent CdTe quantum dots (QDs) were synthesized through a co-precipitation route in aqueous salt solutions using different thiols as stabilizers. The synthetic procedure was simple, efficient, and stable. It could also allow controlling the emission wavelength by varying the experimental conditions such as reaction time and pH values. The strong luminescence of the QDs was observed under UV-excitation and emission colors could be adjusted. The interaction between CdTe QDs and triethylenetetramine dihydrochloride (TETA) which is a candidate treatment for diabetic cardiovascular complication was investigated by fluorescence spectroscopy. Based on the quenching effect on CdTe photoluminescence intensity by TETA, a simple assay system for analyzing the content of TETA in aqueous samples was developed. The linearity was maintained in the range of 0.2 µM to 1.2 µM (R2 = 0.994) with a limit of detection (LOD; S/N = 3) at 28 nM. The results showed that CdTe QDs capped with diverse thiols has a potential for the quantitative analysis of TETA in urine samples.

KHG26792 Inhibits Melanin Synthesis in Mel-Ab Cells and a Skin Equivalent Model.

The purpose of this study is to characterize the effects of KHG26792 (3-(naphthalen-2-yl(propoxy) methyl)azetidine hydrochloride), a potential skin whitening agent, on melanin synthesis and identify the underlying mechanism of action. Our data showed that KHG26792 significantly reduced melanin synthesis in a dose-dependent manner. Additionally, KHG26792 downregulated microphthalmia-associated transcription factor (MITF) and tyrosinase, the rate-limiting enzyme in melanogenesis, although tyrosinase was not inhibited directly. KHG26792 activated extracellular signal-regulated kinase (ERK), whereas an ERK pathway inhibitor, PD98059, rescued KHG26792-induced hypopigmentation. These results suggest that KHG26792 decreases melanin production via ERK activation. Moreover, the hypopigmentary effects of KHG26792 were confirmed in a pigmented skin equivalent model using Cervi cornus Colla (deer antler glue), in which the color of the pigmented artificial skin became lighter after treatment with KHG26792. In summary, our findings suggest that KHG26792 is a novel skin whitening agent.

Lactobacillus kimchiensis sp. nov., isolated from a fermented food.

A novel bacterium was isolated from a traditional fermented food, kimchi. The morphology, physiology, biochemical properties and 16S rRNA gene sequence of strain L133(T) were studied. Strain L133(T) was Gram-reaction-positive, catalase-negative and homofermentative, with rod-shaped cells that formed cream colonies. Cells grew in the presence of 0-5 % (w/v) NaCl (optimum, 1-2 %), at pH 5.0-9.0 (optimum, pH 7.0-8.0) and at 15-37 °C (optimum, 25 °C). Comparative 16S rRNA gene and pheS sequence analysis of strain L133(T) indicated that the strain belonged to the genus Lactobacillus. The major fatty acids were identified as C18 : 1ω9c, C16 : 0 and C18 : 0, and the cell wall contained peptidoglycan of the L-Lys-D-Asp type. DNA-DNA relatedness values between strain L133(T) and related species were below 11±0.4 %. The DNA G+C content of strain L133(T) was 35.7 mol%. Analysis of 16S rRNA gene sequences, as well as physiological and biochemical tests, identified genotypic and phenotypic differences between strain L133(T) and other species of the genus Lactobacillus. Based on these analyses, strain L133(T) is proposed to be a novel species of the genus Lactobacillus, named Lactobacillus kimchiensis. The type strain is L133(T) ( = KACC 15533(T) = JCM 17702(T) = DSM 24716(T)).

Ruegeria conchae sp. nov., isolated from the ark clam Scapharca broughtonii.

A slightly halophilic, Gram-negative, strictly aerobic, non-motile rod, designated TW15(T), was isolated from an ark clam in South Korea. Growth occurred at 10-37 °C, with 1-5% (w/v) NaCl and at pH 7.0-10.0. Optimal growth occurred at 25-30 °C, with 2% (w/v) NaCl and at pH 8.0. Strain TW15(T) exhibited both oxidase and catalase activities. The major fatty acids of strain TW15(T) were summed feature 8 (consisting of C(18:1)ω7c and/or C(18:1)ω6c) and 11-methyl C(18:1)ω7c. The predominant isoprenoid quinone was ubiquinone-10 (Q-10). The polar lipids of strain TW15(T) comprised phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified phospholipid, an unidentified aminolipid and five unidentified lipids. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain TW15(T) was most closely related to Ruegeria lacuscaerulensis DSM 11314(T) (98.0% 16S rRNA gene sequence similarity). DNA-DNA relatedness with closely related strains was <52 ± 3%. The DNA G+C content was 55.7 mol%. On the basis of phenotypic, genotypic and phylogenetic data, strain TW15(T) represents a novel species of the genus Ruegeria, for which the name Ruegeria conchae sp. nov. is proposed. The type strain is TW15(T) ( = KACC 15115(T)  = JCM 17315(T)).

Draft genome sequence of Dietzia alimentaria 72T, belonging to the family Dietziaceae, isolated from a traditional Korean food.

Actinobacterial strain 72(T), named Dietzia alimentaria, which belongs to the family Dietziaceae, was isolated from a traditional Korean food made from clams. The draft genome sequence of D. alimentaria 72(T) contains 3,352,817 bp, with a G+C content of 67.34%.

Virgibacillus alimentarius sp. nov., isolated from a traditional Korean food.

A novel, Gram-positive, rod-shaped, motile, endospore-forming, halophilic bacterial strain, J18(T), was isolated from a traditional salt-fermented seafood made of gizzard shad in Korea. Colonies were convex, cream-coloured and 1.0-2.0 mm in diameter after incubation for 3 days on marine agar. Growth occurred at pH 7.0-11.0 (optimum, pH 10.0), at 4-40 °C (optimum, 37 °C) and in the presence of 0-30% NaCl (optimum, 9-10%). On the basis of 16S rRNA gene sequence analysis, strain J18(T) was related most closely to Virgibacillus byunsanensis ISL-24(T) (96.3% similarity), Virgibacillus carmonensis LMG 20964(T) (96.2%), Virgibacillus halodenitrificans DSM 10037(T) (96.0%), Virgibacillus arcticus Hal 1(T) (95.5%) and Virgibacillus necropolis LMG 19488(T) (95.5%). The major fatty acids were anteiso-C(15:0) and anteiso-C(17:0). The DNA G+C content of strain J18(T) was 37.0 mol%. The cell-wall peptidoglycan was of the meso-diaminopimelic acid type. The major quinone was menaquinone 7 (MK-7). Based on phenotypic, chemotaxonomic and phylogenetic data, strain J18(T) is considered to represent a novel species of the genus Virgibacillus, for which the name Virgibacillus alimentarius sp. nov. is proposed. The type strain is J18(T) (=KACC 14624(T) =JCM 16994(T)).

Dietzia alimentaria sp. nov., isolated from a traditional Korean food.

An actinobacterial strain, designated 72(T), was isolated from a traditional salt-fermented seafood in Korea. Colonies were coral red and cells were Gram-reaction-positive, non-motile rods. Strain 72(T) grew with 0-10 % (w/v) NaCl, at pH 7-10 and at 15-37 °C. Optimum growth conditions were 2 % NaCl, pH 7.0 and 30 °C. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain 72(T) belonged to the genus Dietzia. The major cellular fatty acids (>5 %) were C16:0, summed feature 3 (comprising C16:1ω6c and/or C16:1ω7c), 10-methyl C18:0, C17:0, C19:0 and C18:1ω9c. 16S rRNA gene sequence analysis and DNA-DNA hybridization, coupled with physiological and biochemical tests, revealed genotypic and phenotypic differences between strain 72(T) and other members of the genus Dietzia. Based on these data, strain 72(T) represents a novel species, for which the name Dietzia alimentaria sp. nov. is proposed. The type strain is 72(T) ( = JCM 16360(T)  = KACC 21126(T)).

Probing the transition state for nucleic acid hybridization using phi-value analysis.

Genetic regulation by noncoding RNA elements such as microRNA and small interfering RNA (siRNA) involves hybridization of a short single-stranded RNA with a complementary segment in a target mRNA. The physical basis of the hybridization process between the structured nucleic acids is not well understood primarily because of the lack of information about the transition-state structure. Here we use transition-state theory, inspired by phi-value analysis in protein folding studies, to provide quantitative analysis of the relationship between changes in the secondary structure stability and the activation free energy. Time course monitoring of the hybridization reaction was performed under pseudo-steady-state conditions using a single fluorophore. The phi-value analysis indicates that the native secondary structure remains intact in the transition state. The nativelike transition state was confirmed via examination of the salt dependence of the hybridization kinetics, indicating that the number of sodium ions associated with the transition state was not substantially affected by changes in the native secondary structure. These results propose that hybridization between structured nucleic acids undergoes a transition state leading to formation of a nucleation complex and then is followed by sequential displacement of preexisting base pairings involving successive small energy barriers. The proposed mechanism might provide new insight into physical processes during small RNA-mediated gene silencing, which is essential to selection of a target mRNA segment for siRNA design.

Self-assembled nucleic acid nanoparticles capable of controlled disassembly in response to a single nucleotide mismatch.

We have demonstrated the self-assembled DNA nanoparticles capable of controlled disassembly in response to a single nucleotide change (SNC) in a target nucleic acid. The DNA nanoparticles (avg diameter=51+/-22 nm) were constructed by joining two types of streptavidin-DNA conjugates with 2 molar equiv of a linker strand that carries complementary sequences to both conjugates. Nanoparticle disassembly triggered by a target strand (i.e., a perfect complement to the linker) selectively over mismatched targets was achieved by kinetically controlled nucleation occurring at a 6-nt overhang in the linker. The disassembly process was shown to be dramatically slowed down when using mismatch targets in which the SNC was positioned at the fourth nucleotide from the 3'-end. To verify whether the controlled disassembly also works for a SNC located in the middle of a target strand, we tested a deleterious Z variant (G1024A) of human alpha(1)-antitrypsin as a mismatch target (60-nt) carrying the point mutation at position 39. The wild-type target completed the disassembly process in less than 10 min, whereas the mismatch Z-type target could not complete the disassembly even in 3 h. The DNA nanoparticles are promising for sequence-dependent controlled release of short nucleic acids, including siRNA and antisense oligonucleotides, and construction of smart nanomaterials capable of sensing and processing single-nucleotide polymorphisms.

Single-fluorophore monitoring of DNA hybridization for investigating the effect of secondary structure on the nucleation step.

Nucleic acid hybridization is one of the essential biological processes involved in storage and transmission of genetic information. Here we quantitatively determined the effect of secondary structure on the hybridization activation energy using structurally defined oligonucleotides. It turned out that activation energy is linearly proportional to the length of a single-stranded region flanking a nucleation site, generating a 0.18 kcal/mol energy barrier per nucleotide. Based on this result, we propose that the presence of single-stranded segments available for non-productive base pairing with a nucleation counterpart extends the searching process for nucleation sites to find a perfect match. This result may provide insights into rational selection of a target mRNA site for siRNA and antisense gene silencing.

Facilitating student learning with critical reflective journaling in psychiatric mental health nursing clinical education: A qualitative study.

BACKGROUND: Critical reflective journaling has been used to facilitate student learning in various disciplines; however, there is only limited data available on its application in clinical education in the area of psychiatric mental health nursing. OBJECTIVES: The aims of the study were to: 1) explore types of events or issues that senior nursing students chose to reflect upon in their critical reflective journals during their 5-week psychiatric mental health nursing clinical practicum; and 2) assess students' evaluations of critical reflective journaling. DESIGN & METHODS: For this qualitative study, we collected data from senior nursing students who were enrolled in the mental health clinical practicum from March to June 2016 and agreed to participate in the study. During the 5-week psychiatric mental health nursing clinical practicum, students completed four weeks of clinical rotations in two units and submitted one journal entry each week reflecting upon an instance they selected and completed a survey to evaluate critical reflective journaling. De-identified data were analyzed using qualitative content analysis. RESULTS: We analyzed 236 journal entries and survey results completed by 59 students. Students chose to reflect upon the following issues: boundaries of therapeutic relationships, responses to patient symptoms, and own attitudes and biases toward mental illness. They reported that critical reflective journaling provided them opportunities for self-reflection and self-emancipation, which led to increased motivation for full engagement in the clinical practicum and increased understanding of patients' perspectives as well as mental health nursing competency. Overall, students were satisfied with critical reflective journaling but also identified areas for improvement in regard to its application in clinical education. CONCLUSIONS: Through critical reflective journaling, nursing students are able to self-reflect on issues arisen during their clinical rotation, which leads to improvement of their competency in psychiatric mental health nursing and full engagement in the clinical practicum.

Effects of a Community Outreach Program for Maternal Health and Family Planning in Tigray, Ethiopia.

PURPOSE: We examined the effects of a community outreach program for maternal health in Tigray, Ethiopia, on women's knowledge about pregnancy, childbirth, postpartum care, and family planning, and assessed their participation in antenatal care, postpartum checkups, institutional childbirth, and contraceptive use. METHODS: We recruited Ethiopian women of reproductive age (15 to 49 years) in Tigray, Ethiopia. Two villages in Tigray, Kihen and Mesanu, which have similar population sizes and living environments, were chosen as the intervention and comparison groups, respectively. A two-group pretest-posttest design with cluster sampling was employed. We conducted self-report questionnaire surveys using face-to-face interviews. The 2.5-year community outreach program was developed based on Rogers' diffusion of innovation theory. It consisted of mass media use and health education for lay women in the community, along with training of health care providers, including nurses, midwives, and health extension workers, in maternal health care. RESULTS: The intervention group showed significant increases in knowledge and behaviors regarding maternal health and family planning compared to the comparison group (p < .001). In particular, there was a dramatic increase from 10.8% to 93.5% in the institutional birth rate in the intervention group. CONCLUSION: The community outreach program and health care professional training effectively improved knowledge and behaviors regarding maternal health in Ethiopian women. Mass media and interpersonal communication channels for health education may be useful health interventions in developing countries.

Adenosine potentiates the therapeutic effects of neural stem cells expressing cytosine deaminase against metastatic brain tumors.

Tumor-tropic properties of neural stem cells (NSCs) provide a novel approach with which to deliver targeting therapeutic genes to brain tumors. Previously, we developed a therapeutic strategy against metastatic brain tumors using a human NSC line (F3) expressing cytosine deaminase (F3.CD). F3.CD converts systemically administered 5-fluorocytosine (5-FC), a blood-brain barrier permeable nontoxic prodrug, into the anticancer agent 5-fluorouracil (5-FU). In this study, we potentiated a therapeutic strategy of treatment with nucleosides in order to chemically facilitate the endogenous conversion of 5-FU to its toxic metabolite 5-FU ribonucleoside (5-FUR). In vitro, 5-FUR showed superior cytotoxic activity against MDA-MB-435 cancer cells when compared to 5-FU. Although adenosine had little cytotoxic activity, the addition of adenosine significantly potentiated the in vitro cytotoxicity of 5-FU. When MDA-MB­435 cells were co-cultured with F3.CD cells, F3.CD cells and 5-FC inhibited the growth of MDA-MB-435 cells more significantly in the presence of adenosine. Facilitated 5-FUR production by F3.CD was confirmed by an HPLC analysis of the conditioned media derived from F3.CD cells treated with 5-FC and adenosine. In vivo systemic adenosine treatment also significantly potentiated the therapeutic effects of F3.CD cells and 5-FC in an MDA-MB-435 metastatic brain tumor model. Simple adenosine addition improved the antitumor activity of the NSCs carrying the therapeutic gene. Our results demonstrated an increased therapeutic potential, and thereby, clinical applicability of NSC-based gene therapy.

Effects of Cervi cornus Colla (deer antler glue) in the reconstruction of a skin equivalent model.

The aim of this study was to investigate the effects of Cervi cornus Colla (CCC) in the reconstruction of skin equivalent (SE). H&E staining showed that SE containing hyaluronic acid (HA) or HA and CCC had a thicker epidermis than the control SE. Immunohistochemical staining showed that p63 was mainly present at the basal layer of the epidermis in the HA and CCC model. Involucrin was obviously expressed in the upper layer of the epidermis in the HA and CCC model. Moreover, we observed that integrins α6 and ß1 were strongly expressed along the basement membrane zone in the HA and CCC model, in which the dermis expressing type I collagen was more compact. In conclusion, our data indicate that CCC contributed to the formation of epidermis, basement membrane, and extracellular matrix in the reconstruction of SE and suggest that CCC may be a useful adjuvant in the reconstruction of SE.

Genetically engineered human neural stem cells with rabbit carboxyl esterase can target brain metastasis from breast cancer.

Neural stem cells (NSCs) led to the development of a novel strategy for delivering therapeutic genes to tumors. NSCs expressing rabbit carboxyl esterase (F3.CE), which activates CPT-11, significantly inhibited the growth of MDA-MB-435 cells in the presence of CPT-11. F3.CE cells migrated selectively into the brain metastases located in the opposite hemisphere. The treatment also significantly decreased tumor volume in immune-deficient mice bearing MDA-MB-435 tumors when F3.CE cells were transplanted into the contralateral hemisphere. The survival rate was significantly prolonged with the treatment with F3.CE and CPT-11. This strategy may be considered as an effective treatment regimen for brain metastases.