Methylsulfonylmethane promotes melanogenesis via activation of JNK in Mel-Ab cells.

OBJECTIVE: Methylsulfonylmethane (MSM), which contains organic sulphur, has been used for a long time as a medicinal ingredient because of its benefits to human health. MSM is reported to be protective against certain skin disorders, but it is unknown whether it affects melanin synthesis. Therefore, in our current research, we examined the possibility of MSM controlling the production of melanin in Mel-Ab melanocytes. METHODS: In Mel-Ab cells, melanin contents and tyrosinase activities were assessed and quantified. The expression of microphthalmia-associated transcription factor (MITF) and tyrosinase was evaluated using western blot analysis, while MSM-induced signalling pathways were investigated. RESULTS: The MSM treatment significantly resulted in a dose-dependent increase in melanin production. Furthermore, MSM elevated melanin-related proteins, including MITF and tyrosinase. However, the rate-limiting enzyme of melanin production, tyrosinase, was not directly influenced by it. Therefore, we investigated potential melanogenesis-related signalling pathways that may have been triggered by MSM. Our findings showed that MSM did not influence the signalling pathways associated with glycogen synthase kinase 3ß, cAMP response-element binding protein, extracellular signal-regulated kinase, or p38 mitogen-activated protein kinase. However, MSM phosphorylated c-Jun N-terminal kinases/stress-activated protein kinase (JNK/SAPK), which is known to induce melanogenesis. SP600125, a specific JNK inhibitor, inhibited MSM-induced melanogenesis. CONCLUSION: Taken together, our study indicates that MSM induces melanin synthesis and may serve as a therapeutic option for hypopigmentary skin disorders such as vitiligo.

Leucine rich repeat LGI family member 3: Integrative analyses support its prognostic association with pancreatic adenocarcinoma.

Leucine rich repeat LGI family member 3 (LGI3) is a member of the LGI protein family. Previous studies of our group have reported that LGI3 is expressed in adipose tissue, skin and brain, and serves as a multifunctional cytokine. LGI3 may also be involved in cytokine networks in various cancers. This study aimed to analyze differentially expressed genes in pancreatic adenocarcinoma (PAC) tissues and PAC cohort data in order to evaluate the prognostic role of LGI3. The expression microarray and the PAC cohort data were analyzed by bioinformatic methods for differential expression, protein-protein interactions, functional enrichment and pathway analyses, gene co-expression network analysis, and prognostic association analysis. Results showed that LGI3 expression was significantly reduced in PAC tissues. Nineteen upregulated genes and 31 downregulated genes in PAC tissues were identified as LGI3-regulated genes. Protein-protein interaction network analysis demonstrated that 92% (46/50) of the LGI3-regulated genes that were altered in PACs belonged to a protein-protein interaction network cluster. Functional enrichment and gene co-expression network analyses demonstrated that these genes in the network cluster were associated with various processes including inflammatory and immune responses, metabolic processes, cell differentiation, and angiogenesis. PAC cohort analyses revealed that low expression levels of LGI3 were significantly associated with poor PAC prognosis. Analysis of favorable or unfavorable prognostic gene products in PAC showed that 93 LGI3-regulated genes were differentially associated with PAC prognosis. LGI3 expression was correlated with the tumor-infiltration levels of various immune cells. Taken together, these results suggested that LGI3 may be a potential prognostic marker of PAC.

Massive pulmonary embolism after caesarean section managed with surgical thrombectomy bridged with extracorporeal membrane oxygenation: A case report.

INTRODUCTION: Pulmonary embolism (PE) is a rare but fatal complication in postpartum women. Mortality is as high as 65% in massive PE, in which systemic hypotension persists or circulatory collapse occurs. This case report describes a patient who underwent a caesarean section complicated by massive PE. The patient was managed with early surgical embolectomy and bridged with extracorporeal membrane oxygenation (ECMO). PRESENTATION OF CASE: A 36 years old postpartum patient with an unremarkable medical history had sudden cardiac arrest due to PE on the day after a caesarean section. The patient recovered spontaneous cardiac rhythm after cardiopulmonary resuscitation; however, hypoxia and shock persisted. Cardiac arrest and spontaneous circulation recovery were repeated twice per hour. Veno-arterial (VA) ECMO rapidly improved the patient's condition. Surgical embolectomy was conducted 6 h after the initial collapse by the experienced cardiovascular surgeon. The patient's condition improved rapidly, and was weaned from ECMO on postoperative day three. The patient recovered normal heart function and no pulmonary hypertension was observed on follow-up echocardiography performed 15 months later. DISCUSSION: Timely intervention is important in the management of PE because of its rapid progression. VA ECMO is a useful bridge therapy to prevent derangement and severe organ failure. Surgical embolectomy is appropriate following the use of ECMO in postpartum patients because of the risk of major haemorrhagic complications or intracranial haemorrhage. CONCLUSION: In patients who have undergone caesarean section complicated by massive PE, surgical embolectomy is preferred because of the risk of haemorrhagic complications and their relatively young age.

LGI3 promotes human keratinocyte migration in high-glucose environments by increasing the expression of ß-catenin.

The leucine-rich repeat LGI family member 3 (LGI3) has been reported to regulate various functions in epidermal keratinocytes. In this study, we investigated the effects of LGI3 on keratinocyte migration in environments with different glucose concentrations. Our results showed that cell migration is markedly impaired in high-glucose environments compared to in low-glucose environments (control). Nevertheless, the use of LGI3 in high-glucose environments restores cell migration to the normal level. Therefore, we performed LGI3 knockdown to identify the role of LGI3 in cell migration. It was observed that transfecting LGI3 siRNA into HaCaT cells reduces the expression of LGI3 and inhibits wound closure. These results indicate that LGI3 is deeply involved in wound healing in high-glucose environments. Western blot analysis showed that in high-glucose environments, LGI3 increases the phosphorylation of Akt, forkhead box protein O1, and focal adhesion kinase. However, no change was observed in the levels of glycogen synthase kinase 3ß, c-Jun N-terminal kinase, extracellular signal-regulated kinase, or p38 mitogen-activated protein kinase. Further results showed that LY294002, a specific inhibitor of phosphatidylinositol 3-kinase, reduced LGI3-induced cell migration. It is generally known that Akt activation leads to the accumulation of ß-catenin, an important mediator of keratinocyte migration. LGI3 greatly increased the expression of ß-catenin in high-glucose environments comparison to that in the low-glucose environments. Taken together, these data indicate that LGI3 induces keratinocyte migration in high-glucose environments as a result of ß-catenin accumulation via Akt phosphorylation. Therefore, LGI3 can be considered a new treatment option for diabetic wound healing.

Development of an End-of-Life Nursing Care Protocol for Intensive Care Units: Delphi Survey Method.

Because of the lack of guidelines and standardized protocols for end-of-life nursing care in intensive care units in Korea, many nurses have reported facing difficulties when providing care for patients. This study attempted to develop a standardized end-of-life nursing protocol for use in intensive care units. A draft of the end-of-life nursing care protocol was developed after a literature review. A Delphi survey was carried out twice with 30 experts, and the content validity ratio of the items was investigated. The draft end-of-life nursing care protocol was divided into 3 separate stages with 24 items: 8 initial end-of-life care assessment items, 5 ongoing end-of-life care items, and 11 post-end-of-life care items. The content validity ratios of the first and second rounds were 0.33 or greater in each category, demonstrating the validity of the proposed draft as a standardized protocol. Furthermore, at the suggestion of the experts, an extra item was added in the last stage, resulting in 25 items. The results of this study are expected to help leading hospitals in South Korea outline the roles and range of tasks for end-of-life care in an intensive care unit and thereby resolve difficulties for nurses. Furthermore, this will improve the medical services that family members receive during end-of-life care.

Integrative proteomic network analyses support depot-specific roles for leucine rich repeat LGI family member 3 in adipose tissues.

LGI family member 3 (LGI3) is a member of the LGI protein family. In our previous studies, LGI3 was determined to be expressed in adipose tissues, skin and the brain, where it served as a pleiotropic cytokine. The results indicated that LGI3 levels are increased in adipose tissues of obese individuals in comparison with control individuals and that LGI3 suppressed adipogenesis via its receptor, disintegrin and metalloproteinase domain-containing protein 23. Additionally, it was reported that LGI3 upregulates tumor necrosis factor-α and downregulated adiponectin and hypothesized that LGI3 may act as a proinflammatory adipokine involved in adipose tissue inflammation. In the present study, cytokine arrays were used to analyze cytokine levels in adipose tissues and plasma of LGI3-knockout mice and signaling protein arrays used to analyze the expression and phosphorylation of these proteins in LGI3-treated preadipocytes. The results suggested that expression levels of 129 gene products (24 cytokines and 105 signaling proteins) were altered in response to LGI3 deficiency or LGI3 treatment, respectively. Protein-protein interaction network analysis of LGI3-regulated gene products revealed that 94% of the gene products (21 cytokines and 100 signaling proteins) formed an interaction network cluster. Functional enrichment analysis for the LGI3-regulated gene products, including those from our previous studies, revealed an association with numerous biological processes, including inflammatory responses, cellular differentiation and development and metabolic regulation. Gene co-expression network analysis revealed that these LGI3-regulated gene products were involved in various biological processes in an overlapping and differential manner between subcutaneous and visceral adipose tissues. Notably, inflammatory responses were more strongly associated with the LGI3-regulated gene co-expression network in visceral adipose tissues than in subcutaneous adipose tissues. Analysis of expression quantitative trait loci identified four single nucleotide variants that affect expression of LGI3 in an adipose depot-specific manner. Taken together, the results suggested that LGI3 may serve depot-specific roles as an adipokine in adipose tissues.

Efficacy of horse oil on lipopolysaccharide-induced inflammation in human keratinocyte.

OBJECTIVE: To investigate the efficacy of horse oil on lipopolysaccharide (LPS)-induced inflammation in human keratinocytes. METHODS: Western blot analysis was performed to measure the expression of cyclooxygenase-2 (COX-2) and IκBα. ELISA was used to analyze prostaglandin E2 (PGE2) levels. RESULTS: Horse oil decreased LPS-induced COX-2 and PGE2 levels in a dose-dependent manner. Nuclear factor-kappa B (NF-κB) plays a key role in the expression of inflammatory cytokines and mediators. Therefore, we investigated the influence of horse oil on the NF-κB signaling pathways. Horse oil inhibited translocation of NF-κB from the cytosol to the nucleus. Furthermore, LPS-induced degradation of IκBα was recovered by horse oil. The activation of p38 mitogen-activated protein kinase (MAPK) reportedly induces degradation of IκBα In agreement with this, LPS activated p38 MAPK and caused IκBα degradation. Conversely, horse oil inhibited LPS-induced p38 MAPK activation and IκBα degradation. In addition, a specific p38 MAPK inhibitor, SB203580, blocked IκBα degradation. CONCLUSION: Horse oil decreased COX-2 and PGE2 by inhibiting p38 MAPK activation, IκBα degradation, and the translocation of NF-κB.

Differential Morphological, Structural and Biological Characteristics of Cysts in Heterodera Species in Korea.

Morphological (cyst shape, color, and sizes [length (L), maximum width (W), volume and "a" (L/W)]), structural (vulvar cone slope angle [VCSA], surface wrinkle [VCSW], cyst wall thickness, composition, and texture) and biological characteristics (fecundity, hatching, and emergence [number of second-stage juveniles (J2) from a cyst]) in preceding Heterodera glycines (Hg), currently-recorded H. sojae (Hs) and H. trifolii (Ht) were examined by microscopy. Cysts were lemon-shaped, indicating the genus is Heterodera except for Hs that formed frequently globular cysts with significantly flatter VCSA (102.2°) with smooth VCSW than Hg (50.6°) and Ht (82.0°), but not genus Globodera because of the presence of vulvar cone in Hs. Ht was significantly larger in all morphological characteristics than Hg and Hs, suggesting Ht may be diagnosed differentially by cyst sizes and also host plant preferences. Hs showed smaller "a" value with more globular shape and stronger structures with more thickened and strengthened collagen-like texture of cyst wall than Hg and Ht. This suggests Hs may be diagnosed differently by structural characteristics from the others, especially Hg with similar cyst sizes. There were no significant differences in emergence (inoculum potential) among cyst nematodes due to the offset of fecundity and hatching rate; however, the inoculum potential of Hs may be not so persistent as Hg and Ht in fields because of its lower fecundity and higher hatching rate (causing rapid inoculum loss) than the others. These characteristics of cysts provide information useful for simple and differential diagnoses and reliable management of cyst nematodes.

Navigating the Healthy Context Paradox: Identifying Classroom Characteristics that Improve the Psychological Adjustment of Bullying Victims.

The healthy context paradox-an unexpected pattern in which victims' psychological adjustment worsens as the overall level of victimization in a classroom or school declines-implies that reducing the frequency of bullying or victimization incidents does not do enough to help victims of bullying. In light of this finding, it is imperative to identify protective factors that alleviate victimization-related distress in the peer ecology. The current study examines classroom-level peer victimization and peer-defending behaviors as moderators of the association between individual-level victimization and psychological adjustment. These classroom-level moderators were tested with a sample of 1373 adolescents (40% girls, Mage: 14 years) from 54 classrooms in South Korean middle schools. Consistent with past findings documenting the healthy context paradox, the results of multilevel modeling indicated that victimized youth experienced a lower level of depressive symptoms in classrooms where victimization was more common. Most importantly, bullied students reported fewer depressive symptoms, on average, in classrooms with relatively high levels of bully-oriented (i.e., confronting the bully), rather than victim-oriented (i.e., comforting the victim), defending behavior. These findings provide a more nuanced understanding of the role of peers' defending behaviors toward bullied adolescents and have significant implications for anti-bullying interventions.

Overexpressed p32 localized in the endoplasmic reticulum and mitochondria negatively regulates calcium­dependent endothelial nitric oxide synthase activit.

The p32 protein plays a crucial role in the regulation of cytosolic Ca2+ concentrations ([Ca2+]c) that contributes to the Ca2+­dependent signaling cascade. Using an adenovirus and plasmid p32­overexpression system, the aim of the study was to evaluate the role of p32 in the regulation of [Ca2+] and its potential associated with Ca2+­dependent endothelial nitric oxide synthase (eNOS) activation in endothelial cells. Using electron and confocal microscopic analysis, p32 overexpression was observed to be localized to mitochondria and the endoplasmic reticulum and played an important role in Ca2+ translocation, resulting in increased [Ca2+] in these organelles and reducing cytosolic [Ca2+] ([Ca2+]c). This decreased [Ca2+]c following p32 overexpression attenuated the Ca2+­dependent signaling cascade of calcium/calmodulin dependent protein kinase II (CaMKII)/AKT/eNOS phosphorylation. Moreover, in aortic endothelia of wild­type mice intravenously administered adenovirus encoding the p32 gene, increased p32 levels reduced NO production and accelerated reactive oxygen species (ROS) generation. In a vascular tension assay, p32 overexpression decreased acetylcholine (Ach)­induced vasorelaxation and augmented phenylephrine (PE)­dependent vasoconstriction. Notably, decreased levels of arginase II (ArgII) protein using siArgII were associated with downregulation of overexpressed p32 protein, which contributed to CaMKII­dependent eNOS phosphorylation at Ser1177. These results indicated that increased protein levels of p32 caused endothelial dysfunction through attenuation of the Ca2+­dependent signaling cascade and that ArgII protein participated in the stability of p32. Therefore, p32 may be a novel target for the treatment of vascular diseases associated with endothelial disorders.

New Approaches to Defender and Outsider Roles in School Bullying.

Using latent profile analysis, this study refined the traditional defender and outsider roles in bullying research and examined intrapersonal, interpersonal, and contextual factors associated with subtypes of roles. Participants were 1,373 adolescents (40% girls, Mage : 14 years) from 54 classrooms in six middle schools. The results revealed that defenders could be classified as either assertive or comforting and that outsiders could be classified as either sympathetic or indifferent. These different profiles were explained by affective empathy, antibullying attitudes, self-efficacy, and responsibility to intervene at the intrapersonal level; popularity and peer preference at the interpersonal level; and peers' antibullying attitudes and expectations at the contextual level. Implications for studying participant roles in bullying research and for advancing antibullying interventions were discussed.

LGI3 is secreted and binds to ADAM22 via TRIF-dependent NF-κB pathway in response to LPS in human keratinocytes.

Recently, we reported that HaCaT human keratinocytes secreted leucine-rich repeat LGI family member 3 (LGI3) protein after exposure to ultraviolet B (UVB) irradiation. In the present study, we aimed to determine whether LGI3 is also released in response to stimulation by lipopolysaccharides (LPS), membrane components of gram-negative bacteria. Our results showed that LGI3 was indeed secreted by LPS-stimulated HaCaT cells. We also found that LPS potently stimulated the induction of cycloxygenase-2 (COX-2), which is involved in the inflammatory response. In addition, LPS-induced LGI3 secretion and COX-2 expression were blocked by NS-398, a selective COX-2 inhibitor. Moreover, LPS activated nuclear factor-κB (NF-κB) via a TRIF-dependent pathway, and activated NF-κB led to LGI3 production in HaCaT cells. For the first time, we predicted the LGI3 promoter sequence and demonstrated that NF-κB bound to the LGI3 gene promoter region. LPS treatment also increased the expression of a disintegrin and metalloproteinase domain-containing protein 22 (ADAM22), a candidate LGI3 receptor. Furthermore, co-immunoprecipitation, flow cytometry, and immunocytochemistry revealed that LGI3 associated with ADAM22 in LPS-treated keratinocytes. Thus, ADAM22 may be an LGI3 receptor in human keratinocytes. Taken together, these data suggest that the TRIF-dependent pathway is a novel regulator of LGI3 secretion in response to LPS stimulation in HaCaT cells and that keratinocyte-derived LGI3 interacts with ADAM22 and mediates LPS-induced inflammation.

Leucine rich repeat LGI family member 3: Integrative analyses reveal its prognostic association with non-small cell lung cancer.

Leucine rich repeat LGI family member 3 (LGI3) is a member of the LGI protein family. Our previous studies reported that LGI3 was expressed in adipose tissues, brain and skin, where it served roles as a multifunctional cytokine and pro-inflammatory adipokine. It was hypothesized that LGI3 may be involved in cytokine networks in cancer. The present study aimed to analyze differentially expressed genes in non-small cell lung cancer (NSCLC) tissues and NSCLC cohort data, to evaluate the prognostic role of LGI3. Expression microarray and NSCLC cohort data were statistically analyzed by bioinformatic methods, and protein-protein interactions, functional enrichment and pathway, gene coexpression network (GCN) and prognostic association analyses were performed. The results demonstrated that the expression levels of LGI3 and its receptor a disintegrin and metalloproteinase domain-containing protein 22 were significantly decreased in NSCLC tissues. A total of two upregulated genes and 11 downregulated genes in NSCLC tissues were identified as LGI3-regulated genes. Protein-protein interaction network analysis demonstrated that all LGI3-regulated genes that were altered in NSCLC were involved in a protein-protein interaction network cluster. Functional enrichment, Kyoto Encyclopedia of Genes and Genomes pathway and GCN analyses demonstrated the association of these genes with the immune and inflammatory responses, angiogenesis, the tumor necrosis factor pathway, and chemokine and peroxisome proliferator-activated receptor signaling pathways. Analysis of NSCLC cohorts revealed that low expression levels of LGI3 was significantly associated with poor prognosis of NSCLC. Analysis of the somatic mutations of the LGI3 gene in NSCLC revealed that the amino acid residues altered in NSCLC included two single nucleotide polymorphism sites and three phylogenetically coevolved amino acid residues. Taken together, these results suggest that LGI3 may be a potential prognostic marker of NSCLC.

Too tough at the top: Using latent class growth analysis to assess cool status during middle school.

INTRODUCTION: Obtaining and maintaining high social status in one's peer group is often a critical developmental goal during adolescence. The present study investigated factors that predict trajectories of cool status for middle school adolescents as well as how different cool status trajectories affect depressive symptoms. METHODS: The participants were 5,991 adolescents (52% girls) from 26 urban middle schools in California. Using latent class growth analysis, baseline assessment occurred in the fall of sixth grade, and repeated assessments occurred in the spring of sixth, seventh, and eighth grades. RESULTS: Three cool status trajectories were identified: (1) a high ascending cool status group (5% of the sample); (2) a decreasing cool status group (25%); and (3) a maintaining low cool status group (70%). Differences in the three groups were explained by GPA and having a reputation as aggressive at the beginning of middle school and the level of depression at the end of middle school. Those in the high ascending cool status group experienced the most depressive symptoms at the end of 8th grade. CONCLUSIONS: The findings suggest the need for a more nuanced perspective on maintaining cool status during adolescence that considers both its risks and benefits.

LGI3 promotes human keratinocyte differentiation via the Akt pathway.

Leucine-rich repeat LGI family member 3 (LGI3), a member of the LGI family, is a secreted protein that is expressed not only in the brain and adipose tissues, but also in various skin cells. We previously reported that LGI3 was secreted after exposure to ultraviolet B and promoted the migration of HaCaT human keratinocytes. In the present study, we investigated whether LGI3 influences the differentiation of keratinocytes. The results show that the expression of involucrin, a keratinocyte differentiation marker, was reduced in tissue from LGI3-knockout mice. Those results indicate that LGI3 plays an important role in keratinocyte differentiation. Therefore, we treated HaCaT cells with LGI3 to examine its effect on keratinocyte differentiation. Protein levels of various differentiation markers were enhanced by treatment with LGI3. Furthermore, expression of differentiation markers was inhibited when keratinocytes were transfected with an siRNA for LGI3. LGI3 strongly activated Akt, whereas it had no apparent effect on extracellular signal-regulated kinase, p38 mitogen-activated protein kinase, or the c-Jun N-terminal kinase. A specific inhibitor of phosphoinositide 3-kinase, LY294002, reduced LGI3-induced expression of differentiation markers in HaCaT cells. Taken together, these results suggest that LGI3 promotes keratinocyte differentiation and could be used as a therapeutic agent to recover skin barrier function in epidermal barrier disruption.

Defending Victims of Bullying in Early Adolescence: A Multilevel Analysis.

Adolescents' defending behaviors in school bullying situations is likely determined by individual characteristics, social status variables, and classroom/school contextual factors operating simultaneously in the peer ecology. However, there is little research on defending behavior that utilizes this multilevel approach. This study investigated how students' willingness to defend victims of bullying was affected by feelings of empathy, perceived popularity, and classroom-level perceived prosocial norms. Participants were 1373 adolescents (40% girls, Mage: 14 yrs) from 54 classrooms in six middle schools in South Korea. These youth reported on their feelings of empathy and how prosocial they perceived their classmates to be. Peer-ratings and peer nominations were used to estimate defending behaviors and which students were perceived as popular. Multilevel analyses showed that participants were more likely to defend victims when they had greater empathy and perceived popularity and when classroom-level prosocial norms were higher. The findings have implications for interventions to reduce school bullying and for studying defending behavior in multiple cultural contexts.

Laminin peptide YIGSR enhances epidermal development of skin equivalents.

Since the use of animal experimentation is restricted with regard to cosmetic materials, alternative in vitro models such as skin equivalents (SEs) are needed. Laminin is one of the major non-collagenous glycoproteins. The pentapeptide YIGSR (Tyr-Ile-Gly-Ser-Arg) is a functional motif of laminin that binds to the laminin receptor. In the present study, we examined whether YIGSR could improve the reconstruction of SEs. YIGSR has no effects on monolayer cell proliferation of CCD25-Sk fibroblasts or HaCaT keratinocytes. Interestingly, YIGSR decreased TGF-ß1 levels, although it promoted type Ι collagen synthesis in CCD25-Sk cells. In HaCaT cells, YIGSR decreased the expression of involucrin and loricrin, which are differentiation markers. Furthermore, YIGSR increased levels of proliferating cell nuclear antigen (PCNA), p63, and integrin α6, and decreased involucrin in SE models. In addition, two models containing YIGSR (mixed with dermal equivalents or added into media) did not show any differences in expression levels of PCNA, p63, integrin α6, and involucrin. Therefore, YIGSR is a useful agent for reconstruction of SEs, independent of its method of application. These results indicate that YIGSR stimulates epidermal proliferation and basement membrane formation while inhibiting keratinocyte differentiation of SEs. Taken together, these results indicate that YIGSR promotes the reconstruction of SEs, potentially via decreased TGF-ß1 levels and consequent inhibition of epidermal differentiation.

Leucine-rich glioma inactivated 3: Integrative analyses reveal its potential prognostic role in cancer.

Leucine-rich glioma inactivated 3 (LGI3) is a secreted protein in vertebrates, which belongs to the LGI family. In our previous study, LGI3 was found to be expressed in brain, adipose tissues and the skin, where it functions as a multifunctional cytokine. In the present study, we used bioinformatic tools to perform data mining, phylogenetics and prognostic association analysis to investigate the prognostic role of LGI3 in cancers. The sequences of LGI3 orthologues were analyzed from various species, and it was found that LGI3 was highly conserved in mammals and that the subsets of amino acid residues were phylogenetically coevolved in four major clusters. Single nucleotide polymorphisms (SNPs) of the human LGI3 gene included 228 functionally relevant variants (missense, nonsense and frameshift) in a total of 1,042 SNPs. Four missense SNPs had a global minor allele frequency ≥0.001. Somatic mutations in cancer with functional relevance were found in various types of cancer, including uterine, stomach and lung cancer. In addition, five amino acid residues with cancer mutations were shown to be coevolved in the vertebrate phylogeny, suggesting their importance in protein dysfunctions in cancer. One conserved amino acid and three SNPs were found to be mutated in stomach cancer and melanoma. Analysis of expression microarray data demonstrated that the expression of LGI3 was significantly associated with the prognosis of brain, colorectal and lung cancer. Taken together, these results suggested that the genetic variations and expression levels of LGI3 have potential value in cancer prognosis.

Avian Collagen Is Useful for the Construction of Skin Equivalents.

As a result of restrictions on animal experimentation, improved skin equivalents (SEs) are needed as alternative test models. This work investigated the effects of avian collagen on the construction of SEs, and to the best of our knowledge is the first study to do so. Hematoxylin and eosin and immunohistochemical staining were used to analyze the SEs. In models containing avian collagen as a dermal equivalent (DE) ingredient, fibroblast proliferation increased by about 60% relative to the control model. Immunohistochemical staining showed that the expression of proliferating cell nuclear antigen (PCNA) and p63 increased in the avian collagen models, while the expression of involucrin, integrin α6, and integrin ß1 remained unchanged. Next, DEs were cryopreserved to allow the easier creation of SEs. Keratinocytes were seeded on thawed DEs, and SEs were constructed. Avian collagen increased the viability of DEs relative to the control. Furthermore, avian collagen increased the expression of PCNA and p63 in keratinocytes on thawed DEs. The results indicate that DEs containing avian collagen can be thawed as needed after cryopreservation. Avian collagen can improve the construction of SEs and be used as part of a dermal kit for SE construction.

Leucine-rich glioma inactivated 3: integrative analyses support its prognostic role in glioma.

BACKGROUND: Leucine-rich glioma inactivated 3 (LGI3) is a secreted protein member of LGI family. We previously reported that LGI3 was expressed in brain, adipose tissues and skin, where it played roles as a multifunctional cytokine. We postulated that LGI3 may be involved in cytokine network in cancers. AIM: This study aimed to analyze differentially expressed genes in glioma tissues and glioma cohort data to investigate the prognostic role of LGI3 and its receptors. MATERIALS AND METHODS: Expression microarray data from Gene Expression Omnibus and glioma cohort data were analyzed using bioinformatic tools for statistical analysis, protein-protein interactions, functional enrichment and pathway analyses and prognostic association analysis. RESULTS: We found that LGI3 and its receptors, ADAM22 and ADAM23, were significantly downregulated in glioma tissues. Eleven upregulated genes and two downregulated genes in glioma tissues were found to be the previously reported LGI3-regulated genes. Protein-protein interaction network analysis showed that 85% of the LGI3-regulated and glioma-altered genes formed a cluster of interaction network. Functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses revealed the association of these genes with hypoxia responses, p53 and Akt signaling and various cancer-related pathways including glioma. Analysis of expression microarray data of glioma cohorts demonstrated that low expression levels of LGI3, ADAM22 and ADAM23 were significantly associated with poor prognosis of glioma. CONCLUSION: These results propose that LGI3 and its receptors may play a prognostic role in glioma.