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The effect of temperature and storage time on DNA integrity after freeze-drying sperm from individuals with normozoospermia

  • Farzaneh Mohammadzadeh Kazorgah;Azam Govahi;Ali Dadseresht;Fatemeh Nejat Pish Kenari;Marziyeh Ajdary;Rana Mehdizadeh;Roya Derakhshan;Mehdi Mehdizadeh
    • Clinical and Experimental Reproductive Medicine
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    • v.51 no.1
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    • pp.42-47
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    • 2024
  • Objective: This study evaluated the effects of temperature and storage time on the quality and DNA integrity of freeze-dried sperm from individuals with normozoospermia. Methods: Normal sperm samples from 15 men aged 24 to 40 years were studied. Each sample was divided into six groups: fresh, freezing (frozen in liquid nitrogen), freeze-dried then preserved at room temperature for 1 month (FD-1m-RT), freeze-dried then preserved at room temperature for 2 months (FD-2m-RT), freeze-dried then preserved at 4 ℃ for 1 month (FD-1m-4 ℃), and freeze-dried then preserved at 4 ℃ for 2 months (FD-2m-4 ℃). The morphology, progressive motility, vitality, and DNA integrity of the sperm were evaluated in all groups. Results: In all freeze-dried groups, sperm cells were immotile after rehydration. The freeze-dried groups also showed significantly less sperm vitality than the fresh and frozen groups. Significantly more morphological sperm abnormalities were found in the freeze-dried groups, but freeze-drying did not lead to a significantly higher DNA fragmentation index (DFI). The DFI was significantly higher in the FD-2m-RT group than in the other freeze-dried groups. Conclusion: The freeze-drying method preserved the integrity of sperm DNA. The temperature and duration of storage were also identified as factors that influenced the DFI. Accordingly, more research is needed on ways to improve sperm quality in the freeze-drying process.

What are the Appropriate Surgery and Postoperative Surveillance for Intraductal Papillary Mucinous Neoplasm?

  • Ideno, Noboru;Nakata, Kohei;Nakamura, Masafumi
    • Journal of Digestive Cancer Research
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    • v.9 no.1
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    • pp.8-18
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    • 2021
  • Although many guidelines for pancreatic cystic neoplasms focus on the management of intraductal papillary mucinous neoplasm of the pancreas (IPMN) at the highest oncological risk, there are many issues that surgeons need to consider at the time to plan the surgical procedures based on characteristics of IPMN subtypes, such as multiplicity of branch duct-IPMN (BD-IPMN) and intraductal spreading of main duct-IPMN (MD-IPMN). For multifocal BD-IPMN, partial pancreatectomy would be selected to remove BD-IPMN with predictors of malignancy, while the other lesions without risk factors can be left, although total pancreatectomy might be considered if the patients have a strong family history of pancreatic cancer. Partial pancreatectomy would be also adequate procedure for MD-IPMN if negative surgical margin for high-grade dysplasia or invasive carcinoma were achieved. It has become to be well-known that patients with BD-IPMN are at increased risk for developing not only IPMN-associated pancreatic ductal adenocarcinoma (PDAC) but also PDAC independent from the IPMN. Hence, the detection of a concomitant PDAC is also an important focus for strategies after resection of BD-IPMNs. Our recent analysis of patients after partial pancreatectomy for MD-IPMN with negative surgical margin identified an unexpected recurrence pattern, which we called "monoclonal skip" recurrence. MD-IPMN seems to be disseminated in the pancreatic ductal systems and MD-IPMN with identical genetic background was detected in the remnant pancreas even in a long time after index surgery. We proposed strategies of postoperative surveillance based on characteristics and natural history of each morphological subtype.

Virtual Screening and Testing of GSK-3 Inhibitors Using Human SH-SY5Y Cells Expressing Tau Folding Reporter and Mouse Hippocampal Primary Culture under Tau Cytotoxicity

  • Chih-Hsin Lin;Yu-Shao Hsieh;Ying-Chieh Sun;Wun-Han Huang;Shu-Ling Chen;Zheng-Kui Weng;Te-Hsien Lin;Yih-Ru Wu;Kuo-Hsuan Chang;Hei-Jen Huang;Guan-Chiun Lee;Hsiu Mei Hsieh-Li;Guey-Jen Lee-Chen
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.127-138
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    • 2023
  • Glycogen synthase kinase-3β (GSK-3β) is an important serine/threonine kinase that implicates in multiple cellular processes and links with the neurodegenerative diseases including Alzheimer's disease (AD). In this study, structure-based virtual screening was performed to search database for compounds targeting GSK-3β from Enamine's screening collection. Of the top-ranked compounds, 7 primary hits underwent a luminescent kinase assay and a cell assay using human neuroblastoma SH-SY5Y cells expressing Tau repeat domain (TauRD) with pro-aggregant mutation ΔK280. In the kinase assay for these 7 compounds, residual GSK-3β activities ranged from 36.1% to 90.0% were detected at the IC50 of SB-216763. In the cell assay, only compounds VB-030 and VB-037 reduced Tau aggregation in SH-SY5Y cells expressing ΔK280 TauRD-DsRed folding reporter. In SH-SY5Y cells expressing ΔK280 TauRD, neither VB-030 nor VB-037 increased expression of GSK-3α Ser21 or GSK-3β Ser9. Among extracellular signal-regulated kinase (ERK), AKT serine/threonine kinase 1 (AKT), mitogen-activated protein kinase 14 (P38) and mitogenactivated protein kinase 8 (JNK) which modulate Tau phosphorylation, VB-037 attenuated active phosphorylation of P38 Thr180/ Tyr182, whereas VB-030 had no effect on the phosphorylation status of ERK, AKT, P38 or JNK. However, both VB-030 and VB-037 reduced endogenous Tau phosphorylation at Ser202, Thr231, Ser396 and Ser404 in neuronally differentiated SH-SY5Y expressing ΔK280 TauRD. In addition, VB-030 and VB-037 further improved neuronal survival and/or neurite length and branch in mouse hippocampal primary culture under Tau cytotoxicity. Overall, through inhibiting GSK-3β kinase activity and/or p-P38 (Thr180/Tyr182), both compounds may serve as promising candidates to reduce Tau aggregation/cytotoxicity for AD treatment.

Analysis of Nursing Area Concept Analysis Research Trends over the Last 10 Years (2009-2018): Using Integrated Literature Review (통합적 문헌고찰에 의한 최근 10년(2009∼2018)동안의 국내 간호분야 개념분석 연구 동향 분석)

  • Ahn, Ju-hyun;Jang, Ok-ju;Hyun, Hye-jin;Kim, Joo Hyun;Kim, Young-eun;Lee, Yong-mi;Kim, Myoung-suk
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.4
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    • pp.944-957
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    • 2020
  • This study is an integrated literature review of concept analysis research published in the Journal of the Korean Nursing Society and the journal of branch science for the last 10 years from 2009 to 2018. A total of 30 papers were analyzed, including the metaparadigm of nursing, the type of concept, the method of concept analysis, the degree of utilization, etc. As a result, the metaparadgim of nursing was the most common human being, with 23 human beings, 5 nursing, 1 health, and 1 environment, and all kinds of concepts were identified as abstract concepts. As for concept analysis methods, Walker & Avent had the most number of workers and events with 13 workers, 10 hybrids, and 7 evolutionary methods, but all of them were hybrid methods after 2018. The progress toward tool development has not been confirmed. The diversity of nursing paradigm based on research results and the development of tools based on it are also necessary.

Is Colorectal Cancer A Western Disease? Role of Knowledge and Influence of Misconception on Colorectal Cancer Screening among Chinese and Korean Americans: A Mixed Methods Study

  • Lu, Xiaoxiao;Holt, Cheryl L;Chen, Julia C;Le, Daisy;Chen, Jingjing;Kim, Gil-yong;Li, Jun;Lee, Sunmin
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.11
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    • pp.4885-4892
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    • 2016
  • Background: Chinese and Korean Americans have lower colorectal cancer (CRC) screening rates than other racial/ ethnic groups, which may be explained by a low level of CRC knowledge and a high level of misconceptions. This study explores the role of knowledge in CRC screening among these groups. Methods: Chinese (N=59) and Korean (N=61) Americans older than 50 were recruited from the Washington D.C. Metropolitan area. They completed a detailed survey and participated in focus groups to discuss their knowledge on CRC and CRC screening. Seventeen physicians, community leaders, and patient navigators participated in key informant interviews. Using a mixed methods approach, data were analyzed quantitatively and qualitatively. Results: Participants lacked knowledge about CRC and CRC screening. More than half did not know that screening begins at age 50 and there are several types of tests available. More than 30% thought CRC screening was not necessary if there were no symptoms or there was nothing they could do to prevent CRC. Focus group findings suggested understanding about CRC was limited by an inadequate source of linguistically and culturally relevant health information. For example, many participants considered CRC a western condition mainly caused by unhealthy diet. This led to under-estimations about their susceptibility to CRC. Knowledge was positively associated with self-reported screening. Participants who had higher knowledge scores were more likely to report ever having had a colonoscopy and confidence in ability to have CRC screening. Conclusions: Mixed-methods analysis provides multi-faceted perspectives on CRC knowledge and its influence on screening. Study findings can help inform interventions to increase CRC screening among Chinese and Korean Americans.

Sensitive and Noninvasive Detection of Aberrant SFRP2 and MGMT-B Methylation in Iranian Patients with Colon Polyps

  • Naini, M Alizade;Mokarram, P;Kavousipour, S;Zare, N;Atapour, A;Zarin, M Hassan;Mehrabani, G;Borji, M
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.2185-2193
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    • 2016
  • Background: The pathogenesis of sporadic colorectal cancer (CRC) is influenced by the patient genetic background and environmental factors. Based on prior understanding, these are classified in two major pathways of genetic instability. Microsatellite instability (MSI) and CPG island methylator phenotype (CIMP) are categorized as features of the hypermethylated prototype, and chromosomal instability (CIN) is known to be indicative of the non-hypermethylated category. Secreted frizzled related protein 2 (SFRP2), APC1A in WNT signaling pathway and the DNA repair gene, O6-methylguanine-DNA methyltransferase (MGMT), are frequently hypermethylated in colorectal cancer. Detection of methylated DNA as a biomarker by easy and inexpensive methods might improve the quality of life of patients with CRC via early detection of cancer or a precancerous condition. Aim: To evaluate the rate of SFRP2 and MGMT hypermethylation in both polyp tissue and serum of patients in south Iran as compared with matched control normal population corresponding samples. Materials and Methods: Methylation-specific PCR was used to detect hypermethylation in DNA extracted from 48 polypoid tissue samples and 25 healthy individuals. Results: Of total polyp samples, 89.5% had at least one promoter gene hypermethylation. The most frequent methylated locus was SFRP2 followed by MGMT-B (81.2 and 66.6 percent respectively). Serologic detection of hypermethylation was 95% sensitive as compared with polyp tissue. No hypermethylation was detected in normal tissue and serum and its detection in patients with polyps, especially of serrated type, was specific. Conclusions: Serologic investigation for detection of MGMT-B, SFRP2 hypermethylation could facilitate prioritization of high risk patients for colonoscopic polyp detection and excision.

The activation of NLRP3 inflammasome potentiates the immunomodulatory abilities of mesenchymal stem cells in a murine colitis model

  • Ahn, Ji-Su;Seo, Yoojin;Oh, Su-Jeong;Yang, Ji Won;Shin, Ye Young;Lee, Byung-Chul;Kang, Kyung-Sun;Sung, Eui-Suk;Lee, Byung-Joo;Mohammadpour, Hemn;Hur, Jin;Shin, Tae-Hoon;Kim, Hyung-Sik
    • BMB Reports
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    • v.53 no.6
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    • pp.329-334
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    • 2020
  • Inflammasomes are cytosolic, multiprotein complexes that act at the frontline of the immune responses by recognizing pathogen- or danger-associated molecular patterns or abnormal host molecules. Mesenchymal stem cells (MSCs) have been reported to possess multipotency to differentiate into various cell types and immunoregulatory effects. In this study, we investigated the expression and functional regulation of NLR Family Pyrin Domain Containing 3 (NLRP3) inflammasome in human umbilical cord blood-derived MSCs (hUCB-MSCs). hUCB-MSCs expressed inflammasome components that are necessary for its complex assembly. Interestingly, NLRP3 inflammasome activation suppressed the differentiation of hUCB-MSCs into osteoblasts, which was restored when the expression of adaptor proteins for inflammasome assembly was inhibited. Moreover, the suppressive effects of MSCs on T cell responses and the macrophage activation were augmented in response to NLRP3 activation. In vivo studies using colitic mice revealed that the protective abilities of hUCB-MSCs increased after NLRP3 stimulation. In conclusion, our findings suggest that the NLRP3 inflammasome components are expressed in hUCB-MSCs and its activation can regulate the differentiation capability and the immunomodulatory effects of hUCB-MSCs.

Reticuloendothelial System Potentiating of Polysaccharide from Panax Species (Panax속 식물의 다당류가 망내계 활성에 미치는 영향)

  • Ohtani Kazuhiro;Hirose Kumi;Hatana Shunso;Mizutani Kenji;Kasai Ryoji;Tanaka Osamu;Masuda Hitoshi;Furukawa Hiromi;Fuwa Tohru
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.147-150
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    • 1988
  • Polysaccharides which show reticuloendothelial system potentiating activity in carbon clearance tests have been examined in water extracts of Panx species. From the dried roots of P. notoginseng, an active polysaccharide called sanchinan-A was isolated. The molecular weight of sanchinan-A was estimated to be 1,500,000D and the structure was determined to be $\beta$-D-(1-3-galactan), possessing branch points at positions 0-6 at which (mainly-$\alpha$-L-arabinofuranosyl and partly $\beta$-D-galactopyranosyl)-(1-6)-$\beta$-D-galacto-pyranosyl-(1-3)-$\beta$-D-galactopyranosyl side chains are attached on average, to two of three galactosyl units. From dried rhizomes of P. japonicus, several active polysaccharides were also isolated, and these structuers were also determined. From the dried roots of P. ginseng, several polysaccharides which showed strong activity were isolated. The structures of these compounds are currently under investigation. The polysaccharide fraction (non-dialyzed fraction) of the water extract of red ginseng (steam-dried roots) did not exhibit activity, while the dialyzed fraction potentiated RES. Activity disappeared, however, during the process of separation due to the presence of a substance in the fraction which stabilizes an active substance.

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Mitochondrial OXPHOS genes provides insights into genetics basis of hypoxia adaptation in anchialine cave shrimps

  • Guo, Huayun;Yang, Hao;Tao, Yitao;Tang, Dan;Wu, Qiong;Wang, Zhengfei;Tang, Boping
    • Genes and Genomics
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    • v.40 no.11
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    • pp.1169-1180
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    • 2018
  • Cave shrimps from the genera Typhlatya, Stygiocaris and Typhlopatsa (TST complex) comprises twenty cave-adapted taxa, which mainly occur in the anchialine environment. Anchialine habitats may undergo drastic environmental fluctuations, including spatial and temporal changes in salinity, temperature, and dissolved oxygen content. Previous studies of crustaceans from anchialine caves suggest that they have possessed morphological, behavioral, and physiological adaptations to cope with the extreme conditions, similar to other cave-dwelling crustaceans. However, the genetic basis has not been thoroughly explored in crustaceans from anchialine habitats, which can experience hypoxic regimes. To test whether the TST shrimp-complex hypoxia adaptations matched adaptive evolution of mitochondrial OXPHOS genes. The 13 OXPHOS genes from mitochondrial genomes of 98 shrimps and 1 outgroup were examined. For each of these genes was investigated and compared to orthologous sequences using both gene (i.e. branch-site and Datamonkey) and protein (i.e. TreeSAAP) level approaches. Positive selection was detected in 11 of the 13 candidate genes, and the radical amino acid changes sites scattered throughout the entire TST complex phylogeny. Additionally, a series of parallel/convergent amino acid substitutions were identified in mitochondrial OXPHOS genes of TST complex shrimps, which reflect functional convergence or similar genetic mechanisms of cave adaptation. The extensive occurrence of positive selection is suggestive of their essential role in adaptation to hypoxic anchialine environment, and further implying that TST complex shrimps might have acquired a finely capacity for energy metabolism. These results provided some new insights into the genetic basis of anchialine hypoxia adaptation.

Towards high-accuracy data modelling, uncertainty quantification and correlation analysis for SHM measurements during typhoon events using an improved most likely heteroscedastic Gaussian process

  • Qi-Ang Wang;Hao-Bo Wang;Zhan-Guo Ma;Yi-Qing Ni;Zhi-Jun Liu;Jian Jiang;Rui Sun;Hao-Wei Zhu
    • Smart Structures and Systems
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    • v.32 no.4
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    • pp.267-279
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    • 2023
  • Data modelling and interpretation for structural health monitoring (SHM) field data are critical for evaluating structural performance and quantifying the vulnerability of infrastructure systems. In order to improve the data modelling accuracy, and extend the application range from data regression analysis to out-of-sample forecasting analysis, an improved most likely heteroscedastic Gaussian process (iMLHGP) methodology is proposed in this study by the incorporation of the outof-sample forecasting algorithm. The proposed iMLHGP method overcomes this limitation of constant variance of Gaussian process (GP), and can be used for estimating non-stationary typhoon-induced response statistics with high volatility. The first attempt at performing data regression and forecasting analysis on structural responses using the proposed iMLHGP method has been presented by applying it to real-world filed SHM data from an instrumented cable-stay bridge during typhoon events. Uncertainty quantification and correlation analysis were also carried out to investigate the influence of typhoons on bridge strain data. Results show that the iMLHGP method has high accuracy in both regression and out-of-sample forecasting. The iMLHGP framework takes both data heteroscedasticity and accurate analytical processing of noise variance (replace with a point estimation on the most likely value) into account to avoid the intensive computational effort. According to uncertainty quantification and correlation analysis results, the uncertainties of strain measurements are affected by both traffic and wind speed. The overall change of bridge strain is affected by temperature, and the local fluctuation is greatly affected by wind speed in typhoon conditions.