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Merging the cryptic genera Radicilingua and Calonitophyllum (Delesseriaceae, Rhodophyta): molecular phylogeny and taxonomic revision

  • Wolf, Marion A.;Sciuto, Katia;Maggs, Christine A.;Petrocelli, Antonella;Cecere, Ester;Buosi, Alessandro;Sfriso, Adriano
    • ALGAE
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    • v.36 no.3
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    • pp.165-174
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    • 2021
  • Radicilingua Papenfuss and Calonitophyllum Aregood are two small genera of the family Delesseriaceae that consist of only three and one taxonomically accepted species, respectively. The type species of these genera, Radicilingua thysanorhizans from England and Calonitophyllum medium from the Americas, are morphologically very similar, with the only recognized differences being vein size and procarp development. To date, only other two species were recognized inside the genus Radicilingua: R. adriatica and R. reptans. In this study, we analysed specimens of Radicilingua collected in the Adriatic and Ionian Sea (Mediterranean), including a syntype locality of R. adriatica (Trieste, northern Adriatic Sea), alongside material from near the type locality of R. thysanorhizans (Torpoint, Cornwall, UK). The sequences of the rbcL-5P gene fragment here produced represent the first molecular data available for the genus Radicilingua. Phylogenetic reconstruction showed that the specimens from the Adriatic and Ionian Seas were genetically distinct from the Atlantic R. thysanorhizans, even if morphologically overlapping with this species. A detailed morphological description of the Mediterranean specimens, together with an accurate literature search, suggested that they were distinct also from R. adriatica and R. reptans. For these reasons, a new species was here described to encompass the Mediterranean specimens investigated in this study: R. mediterranea Wolf, Sciuto & Sfriso. Moreover, in the rbcL-5P tree, sequences of the genera Radicilingua and Calonitophyllum grouped in a well-supported clade, distinct from the other genera of the subfamily Nitophylloideae, leading us to propose that Calonitophyllum medium should be transferred to Radicilingua.

Differentiation and identification of ginsenoside structural isomers by two-dimensional mass spectrometry combined with statistical analysis

  • Xiu, Yang;Ma, Li;Zhao, Huanxi;Sun, Xiuli;Li, Xue;Liu, Shuying
    • Journal of Ginseng Research
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    • v.43 no.3
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    • pp.368-376
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    • 2019
  • Background: In the current phytochemical research on ginseng, the differentiation and structural identification of ginsenosides isomers remain challenging. In this paper, a two-dimensional mass spectrometry (2D-MS) method was developed and combined with statistical analysis for the direct differentiation, identification, and relative quantification of protopanaxadiol (PPD)-type ginsenoside isomers. Methods: Collision-induced dissociation was performed at successive collision energy values to produce distinct profiles of the intensity fraction (IF) and ratio of intensity (RI) of the fragment ions. To amplify the differences in tandem mass spectra between isomers, IF and RI were plotted against collision energy. The resulting data distributions were then used to obtain the parameters of the fitted curves, which were used to evaluate the statistical significance of the differences between these distributions via the unpaired t test. Results: A triplet and two pairs of PPD-type ginsenoside isomers were differentiated and identified by their distinct IF and RI distributions. In addition, the fragmentation preference of PPD-type ginsenosides was determined on the basis of the activation energy. The developed 2D-MS method was also extended to quantitatively determine the molar composition of ginsenoside isomers in mixtures of biotransformation products. Conclusion: In comparison with conventional mass spectrometry methods, 2D-MS provides more direct insights into the subtle structural differences between isomers and can be used as an alternative approach for the differentiation of isomeric ginsenosides and natural products.

Induction of pro-inflammatory cytokines by 29-kDa FN-f via cGAS/STING pathway

  • Hwang, Hyun Sook;Lee, Mi Hyun;Choi, Min Ha;Kim, Hyun Ah
    • BMB Reports
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    • v.52 no.5
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    • pp.336-341
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    • 2019
  • The cGAS-STING pathway plays an important role in pathogen-induced activation of the innate immune response. The 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) found predominantly in the synovial fluid of osteoarthritis (OA) patients increases the expression of catabolic factors via the toll-like receptor-2 (TLR-2) signaling pathway. In this study, we investigated whether 29-kDa FN-f induces inflammatory responses via the cyclic GMP-AMP synthase (cGAS)/stimulator of interferon gene (STING) pathway in human primary chondrocytes. The levels of cGAS and STING were elevated in OA cartilage compared with normal cartilage. Long-term treatment of chondrocytes with 29-kDa FN-f activated the cGAS/STING pathway together with the increased level of gamma-H2AX, a marker of DNA breaks. In addition, the expression of pro-inflammatory cytokines, including granulocyte-macrophage colony-stimulating factor (GM-CSF/CSF-2), granulocyte colony-stimulating factor (G-CSF/CSF-3), and type I interferon ($IFN-{\alpha}$), was increased more than 100-fold in 29-kDa FN-f-treated chondrocytes. However, knockdown of cGAS and STING suppressed 29-kDa FN-f-induced expression of GM-CSF, G-CSF, and $IFN-{\alpha}$ together with the decreased activation of TANK-binding kinase 1 (TBK1), interferon regulatory factor 3 (IRF3), and inhibitor protein ${\kappa}B{\alpha}$ ($I{\kappa}B{\alpha}$). Furthermore, NOD2 or TLR-2 knockdown suppressed the expression of GM-CSF, G-CSF, and $IFN-{\alpha}$ as well as decreased the activation of the cGAS/STING pathway in 29-kDa FN-f-treated chondrocytes. These data demonstrate that the cGAS/STING/TBK1/IRF3 pathway plays a critical role in 29-kDa FN-f-induced expression of pro-inflammatory cytokines.

Observation of Gene Edition by the Transient Expression of CRISPR-Cas9 System During the Development of Tomato Cotyledon (Agrobacterium을 이용한 토마토 떡잎에서 CRISPR-Cas9 시스템의 임시발현 시 토마토 떡잎 발달 단계에 따른 유전자교정 효율 변화)

  • Kim, Euyeon;Yang, So Hee;Park, Hyosun;Koo, Yeonjong
    • Korean Journal of Environmental Agriculture
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    • v.40 no.3
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    • pp.186-193
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    • 2021
  • BACKGROUND: Before generating transgenic plant using the CRISPR-Cas9 system, the efficiency test of sgRNAs is recommended to reduce the time and effort for plant transformation and regeneration process. The efficiency of the sgRNA can be measured through the transient expression of sgRNA and Cas9 gene in tomato cotyledon; however, we found that the calculated efficiency showed a large variation. It is necessary to increase the precision of the experiment to obtain reliable sgRNA efficiency data from transient expression. METHODS AND RESULTS: The cotyledon of 11th, 15th, 19th, and 23rd-day-old tomato (Solanum lycopersicum cv. Micro-Tom) were used for expressing CRISPR-Cas9 transiently. The agrobacterium harboring sgRNA for targeting ALS2 gene of tomato was injected through the stomata of leaf adaxial side and the genomic DNA was extracted in 5 days after injection. The target gene edition was identified by amplifying DNA fragment of target region and analyzing with Illumina sequencing method. The target gene editing efficiency was calculated by counting base deletion and insertion events from total target sequence read. CONCLUSION: The CRISPR-Cas9 editing efficiency varied with tomato cotyledon age. The highest efficiency was observed at the 19-day-old cotyledons. Both the median and mean were the highest at this stage and the sample variability was also minimized. We found that the transgene of CRISPR-Cas9 system was strongly correlated with plant leaf development and suggested the optimum cotyledon leaf age for Agrobacterium-mediated transfection in tomato.

The variation of insulin like growth factor 2 maker is associated with growth traits in Thai native (Kradon) pigs

  • Kessara Ampaporn;Rattikan Suwannasing;Pitchayanipa Phongphanich;Supanon Tunim;Monchai Duangjinda
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1350-1356
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    • 2023
  • Objective: This study was conducted to investigate polymorphisms of the melanocortin-4 receptor (MC4R) and insulin like growth factor 2 (IGF2) genes and to evaluate the growth traits affected by such polymorphisms in Thai native (Kradon) pigs. Methods: Blood samples and productive data from 91 Kradon pigs were collected. DNA was extracted and quantified, the IGF2 and MC4R genes were amplified, and the polymerase chain reaction (PCR) produces were digested using the PCR-restriction fragment length polymorphism (PCR-RFLP) technique. Genotyping was performed, and the association between genotypes and growth traits on the birth and weaning weights were evaluated. Results: The IGF2 intron7 g.162G>C variations in Kradon pigs were found in three genotypes: i) GG, ii) GC, and iii) CC. The GG genotype frequency was the highest followed by the GC and CC genotypes. The frequencies of the G and C alleles were 0.703 and 0.297, respectively. The MC4R genotype was found in only one genotype (GG). The IGF2 gene pattern was not associated with birth weight traits, whereas the IGF2 gene pattern was related to the weaning weight trait in Kradon pigs. Pigs with the CC and GC genotypes had higher weaning weights than ones with the GG genotype (p<0.001). Conclusion: Thai native Kradon pigs with the CC and GC genotypes of the IGF2 gene have higher weaning weights than pigs with the GG genotype.

Comparison of the cyclic fatigue resistance of One Curve, F6 Skytaper, Protaper Next, and Hyflex CM endodontic files

  • Charlotte Gouedard;Laurent Pino;Reza Arbab-Chirani;Shabnam Arbab-Chirani;Valerie Chevalier
    • Restorative Dentistry and Endodontics
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    • v.47 no.2
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    • pp.16.1-16.9
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    • 2022
  • Objectives: This study compared the cyclic fatigue resistance of One Curve (C wire) and F6 Skytaper (conventional austenite nickel-titanium [NiTi]), and 2 instruments with thermos-mechanically treated NiTi: Protaper Next X2 (M wire) and Hyflex CM (CM wire). Materials and Methods: Ten new instruments of each group (size: 0.25 mm, 6% taper in the 3 mm tip region) were tested using a rotary bending machine with a 60° curvature angle and a 5 mm curvature radius, at room temperature. The number of cycles until fracture was recorded. The length of the fractured instruments was measured. The fracture surface of each fragment was examined with a scanning electron microscope (SEM). The data were analyzed using one-way analysis of variance and the post hoc Tukey test. The significance level was set at 0.05. Results: At 60°, One Curve, F6 Skytaper and Hyflex CM had significantly longer fatigue lives than Protaper Next X2 (p < 0.05). No statistically significant differences were found in the cyclic fatigue lives of One Curve, F6 Skytaper, and Hyflex CM (p > 0.05). SEM images of the fracture surfaces of the different instruments showed typical features of fatigue failure. Conclusions: Within the conditions of this study, at 60° and with a 5 mm curvature radius, the cyclic fatigue life of One Curve was not significantly different from those of F6 Skytaper and Hyflex CM. The cyclic fatigue lives of these 3 instruments were statistically significantly longer than that of Protaper Next.

Assessment of Resistance Induction in Mungbean against Alternaria alternata through RNA Interference

  • Hira Abbas;Nazia Nahid;Muhammad Shah Nawaz ul Rehman;Tayyaba Shaheen;Sadia Liaquat
    • The Plant Pathology Journal
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    • v.40 no.1
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    • pp.59-72
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    • 2024
  • A comprehensive survey of mungbean-growing areas was conducted to observe leaf spot disease caused by Alternaria alternata. Alternaria leaf spot symptoms were observed on the leaves. Diversity of 50 genotypes of mungbean was assessed against A. alternata and data on pathological traits was subjected to cluster analysis. The results showed that genotypes of mungbean were grouped into four clusters based on resistance parameters under the influence of disease. The principal component biplot demonstrated that all the disease-related parameters (% disease incidence, % disease intensity, lesion area, and % of infection) were strongly correlated with each other. Alt a 1 gene that is precisely found in Alternaria species and is responsible for virulence and pathogenicity. Alt a 1 gene was amplified using gene specific primers. The isolated pathogen produced similar symptoms when inoculated on mungbean and tobacco. The sequence analysis of the internal transcribed spacer (ITS) region, a 600 bp fragment amplified using specific primers, ITS1 and ITS2 showed 100% identity with A. alternata. Potato virus X (PVX) -based silencing vector expressing Alt a 1 gene was constructed to control this pathogen through RNA interference in tobacco. Out of 50 inoculated plants, 9 showed delayed onset of disease. Furthermore, to confirm our findings at molecular level semi-quantitative reverse transcriptase polymerase chain reaction was used. Both phenotypic and molecular investigation indicated that RNAi induced through the VIGS vector was efficacious in resisting the pathogen in the model host, Tobacco (Nicotiana tabacum). To the best of our knowledge, this study has been reported for the first time.

File Type Identification Using CNN and GRU (CNN과 GRU를 활용한 파일 유형 식별 및 분류)

  • Mingyu Seong;Taeshik Shon
    • Journal of Platform Technology
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    • v.12 no.2
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    • pp.12-22
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    • 2024
  • With the rapid increase in digital data in modern society, digital forensics plays a crucial role, and file type identification is one of its integral components. Research on the development of identification models utilizing artificial intelligence is underway to identify file types swiftly and accurately. However, existing studies do not support the identification of file types with high domestic usage rates, making them unsuitable for use within the country. Therefore, this paper proposes a more accurate file type identification model using Convolutional Neural Networks (CNN) and Gated Recurrent Units (GRU). To overcome limitations of existing methods, the proposed model demonstrates superior performance on the FFT-75 dataset, effectively identifying file types with high domestic usage rates such as HWP, ALZ, and EGG. The model's performance is validated by comparing it with three existing research models (CNN-CO, FiFTy, CNN-LSTM). Ultimately, the CNN and GRU based file type identification and classification model achieved 68.2% accuracy on 512-byte file fragments and 81.4% accuracy on 4096-byte file fragments.

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CLINICAL STUDY OF COMPLICATIONS OF ORTHOGNATHIC SURGERY FOR THE DENTOFACIAL DEFORMITIES (악변형환자의 악교정수술시 합병증에 관한 연구)

  • Kim, Yeo-Gab;Lee, Sang-Chull;Lee, Baek-Soo;Kim, Byung-Ju
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.16 no.3
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    • pp.247-258
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    • 1994
  • We got this conclusion from an investigation about complications from 144 cases of 130 patients who were to be searched their personal records, admission chart, clinical laboratory sheet, anesthetic record, consult sheet and radiographic opinion. They had orthognathic surgery for maxillofacial deformity treatment at the department of oral and maxillofacial surgery in dental school of Kyung Hee university for 4 years and 10 months, from March 1989 to December 1993. 1. In the intraoperative phase, by the frequency of complication, blood vessel injury was found the most-22%, and then soft tissue injury, unfavorable osteotomy split, and there were some cases of tooth injury and inappropriate osteotomy. In the mandibular segmental osteotomy, blood vessel injury was found the most frequently-20 cases (27%), soft tissue injury, unfavorable osteotomy split were the second frequent cases, and then unfavorable fragment position was found. In the extraoral vertical ramus osteotomy and Le Fort I osteotomy also, blood vessel injury and nerve injury were found the most. 2. In the postoperative hospitalization phase, by the frequency of complication, hematoma (23%) was happened the most, except for that, lkie the complication that can be happened by adverse reaction of medicine or long hospital life. In the case of SSRO, there were 21 cases (20%) of hematoma, and this wal the most frequently case. In the case of EVRO, hematoma wasn't happened that much-2 case (4%). 3. In the follow up phase, relaps, numbness and TMJ dysfunction were happened. In mandibular surgery, the forward relapse percentage of point B, was 27% when used wire fixation on SSRO, was 15% by miniplate fixation on SSRO and was 7% on EVRO. In the case of SSRO, numbness was kind of high, comparing to ordinary surgery-12 cases(16 There were many difficulties in analyzing this data accurately, Although orthognathic surgery is done many times, only available date is from the "success" stories and data is not consistently recorded for the cases with complications. In this manner, much essential informantion is lost and overlooked. When data is charted including those cases that are seemingly insignificalt, we can have a much clearer understanding and more accurate guide on treatment protocols.

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Taxonomy of Korean Calanthe species and few of its mutants based on AFLP data (AFLP에 의한 한국산 새우난초속 식물과 그의 수종 돌연변이에 대한 분류학적 연구)

  • Srikanth, Krishnamoorthy;Koo, Ja Choon;Ku, Jajung;Choi, Kyung;Park, Kwang-Woo;So, Soonku;Choi, Yong-Gook;Whang, Sung Soo
    • Korean Journal of Plant Taxonomy
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    • v.42 no.3
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    • pp.215-221
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    • 2012
  • Five Korean Calanthe species, C. discolor, C. bicolor, C. sieboldii, C. reflexa, and C. aristulifera, were studied using amplified fragment length polymorphism (AFLP) to assess their taxonomic and genetic relationships. Sixteen accessions belonging to five native Calanthe spp. and mutants with yellow tepal and white lip (YW mutants) were studied. We identified 50 putative markers using AFLP analysis. The results of AMOVA showed that genetic variance was higher between species than within species. Genetic dissimilarity when compared with the rest of the species was the lowest for individuals of the YW mutants and the highest for individuals of C. reflexa. The mutants clustered outside the major group. Calanthe bicolor clustered with C. discolor, suggesting that its genetic composition is closer to that of C. discolor. Though it is suggested to have originated as a result of natural hybridization between C. sieboldii and C. discolor, introgression is likely to have occurred in the direction of C. discolor based on the data of molecular marker, clustering and genetic dissimilarity. Calanthe reflexa and C. aristulifera were genetically the most diverse of the species studied. In conclusion, the results showed that there is genetic diversity in Korean Calanthe species, that C. bicolor introgressed in the direction of C. discolor and that the YW mutants are genetically closer to C. sieboldii.