Uncovering transcriptional reprogramming during callus development in soybean: insights and implications.

Callus, a valuable tool in plant genetic engineering, originates from dedifferentiated cells. While transcriptional reprogramming during callus formation has been extensively studied in Arabidopsis thaliana, our knowledge of this process in other species, such as Glycine max, remains limited. To bridge this gap, our study focused on conducting a time-series transcriptome analysis of soybean callus cultured for various durations (0, 1, 7, 14, 28, and 42 days) on a callus induction medium following wounding with the attempt of identifying genes that play key roles during callus formation. As the result, we detected a total of 27,639 alterations in gene expression during callus formation, which could be categorized into eight distinct clusters. Gene ontology analysis revealed that genes associated with hormones, cell wall modification, and cell cycle underwent transcriptional reprogramming throughout callus formation. Furthermore, by scrutinizing the expression patterns of genes related to hormones, cell cycle, cell wall, and transcription factors, we discovered that auxin, cytokinin, and brassinosteroid signaling pathways activate genes involved in both root and shoot meristem development during callus formation. In summary, our transcriptome analysis provides significant insights into the molecular mechanisms governing callus formation in soybean. The information obtained from this study contributes to a deeper understanding of this intricate process and paves the way for further investigation in the field.

Dynamic Hemitongue Defect Reconstruction With Functional Gracilis Muscle Free Transfer.

BACKGROUND: Because the tongue consists of 26 intrinsic and extrinsic muscles, even hemiglossectomy, which preserves some of the tongue and its muscles, leads to functional morbidity in speech and swallowing. Subsequent reconstruction using a conventional fasciocutaneous flap results in limited functional recovery. This study compared the functional recovery of patients who underwent hemiglossectomy based on the fasciocutaneous free flap with or without dynamic gracilis muscle flap reconstruction. METHODS: Twenty-three patients were included in the study. Reconstruction was achieved using radial forearm and gracilis conjoined flaps (n = 7), gracilis flaps (n = 7), and radial forearm flaps (RFFFs) (n = 9) between 2014 and 2019. Functional outcome data were collected via videofluoroscopic swallowing, speech analysis, and a tongue movement scale. RESULTS: In the conjoined flap group, the lingual range of motion in terms of elevation and defect-side lateralization was superior to that of the RFFF group. Furthermore, the conjoined flap was associated with superior protrusion, elevation, and lateralization (on both sides) than the gracilis-only flap. Patients who underwent conjoined-flap reconstruction had better articulation, intelligence, and dysphagia outcomes than patients who underwent reconstruction with RFFFs or gracilis-only flaps. CONCLUSIONS: Although this was a preliminary study, the findings suggest that using a conjoined free flap with an RFFF and a functional gracilis muscle flap for dynamic hemitongue reconstructions could improve postoperative tongue function.

A Multifactorial Interpretation of a Teenager's Suicide: Based on Krystal's Death in Casual Vacancy.

OBJECTIVES: It is hard to accumulate research data on adolescents' suicide, because friends and family of the suicide completers might be reluctant to share the experience. To overcome the lack of information on adolescent suicide victims, the authors examined the risk and protective factors for adolescents' suicide from a character in a novel. METHODS: Krystal, an adolescent female in the novel The Casual Vacancy by Joanne Rowling, failed to overcome her unfortunate cir-cumstances and committed suicide. The authors analysed Krystal's case based on the guideline for patients with suicidal behaviours to address the complicated situation of her death. RESULTS: Krystal grew up in a poor and dangerous environment. Despite the environmental hardships, she developed ego maturation with affectionate help from Mr Fairbrother, an assistant coach of the Girls' Rowing Team and a parish councillor. The sudden passing away of Mr Fairbrother brought on a crisis of identity for Krystal. In addition, a villainous character raped her and her brother drowned to death, which brought her great sorrow. She felt helpless and committed suicide. CONCLUSION: In spite of many risk factors for suicide, Krystal was able to keep her life with a few protective factors, a younger brother in the home, and a sense of responsibility for the family. After the loss of her brother, however, she collapsed in a moment. Krystal's suicide might not only be a personal choice but a breakdown of the social protection system for the youth.

Dissecting seed pigmentation-associated genomic loci and genes by employing dual approaches of reference-based and k-mer-based GWAS with 438 Glycine accessions.

The soybean is agro-economically the most important among all cultivated legume crops, and its seed color is considered one of the most attractive factors in the selection-by-breeders. Thus, genome-wide identification of genes and loci associated with seed colors is critical for the precision breeding of crop soybeans. To dissect seed pigmentation-associated genomic loci and genes, we employed dual approaches by combining reference-based genome-wide association study (rbGWAS) and k-mer-based reference-free GWAS (rfGWAS) with 438 Glycine accessions. The dual analytical strategy allowed us to identify four major genomic loci (designated as SP1-SP4 in this study) associated with the seed colors of soybeans. The k-mer analysis enabled us to find an important recombination event that occurred between subtilisin and I-cluster B in the soybean genome, which could describe a special structural feature of ii allele within the I locus (SP3). Importantly, mapping analyses of both mRNAs and small RNAs allowed us to reveal that the subtilisin-CHS1/CHS3 chimeric transcripts generate and act as an initiator towards 'mirtron (i.e., intron-harboring miRNA precursor)'-triggered silencing of chalcone synthase (CHS) genes. Consequently, the results led us to propose a working model of 'mirtron-triggered gene silencing (MTGS)' to elucidate a long-standing puzzle in the genome-wide CHS gene silencing mechanism. In summary, our study reports four major genomic loci, lists of key genes and genome-wide variations that are associated with seed pigmentation in soybeans. In addition, we propose that the MTGS mechanism plays a crucial role in the genome-wide silencing of CHS genes, thereby suggesting a clue to currently predominant soybean cultivars with the yellow seed coat. Finally, this study will provide a broad insight into the interactions and correlations among seed color-associated genes and loci within the context of anthocyanin biosynthetic pathways.

Management of Mid-Vault Asymmetry in Secondary Unilateral Cleft Lip-Nose Deformity.

The focus of secondary rhinoplasty for patients with a cleft lip after facial growth has been on correcting nasal tip asymmetry and distorted cleft-side lower lateral cartilage. However, some patients present with mid-vault asymmetry even after secondary rhinoplasty. The authors propose camouflage procedures for patients with a unilateral cleft lip and without symptoms of nasal airway obstruction. In camouflage procedures, autologous cartilage or acellular dermal matrix was used for onlay grafting and placed on the upper lateral cartilage. In this report, case examples are described to illustrate the surgical techniques and results. This method enables the correction of mid-vault asymmetry without the use of an additional septal spreader graft.

Extensive Facial Reconstruction Using Thickness-controlled Perforator Free Flaps.

BACKGROUND: Although small and medium facial defects can be reconstructed with local flaps to ensure skin color and texture matching, extensive facial defects require the application of free flaps, including perforator free flaps. We hereby propose a personalized strategy for facial reconstruction to overcome these limitations, based on the use of free flaps accompanied by local flaps and thickness-controlled perforator flap concept, for extensive facial defects. METHODS: A series of consecutive facial reconstructions were performed from 2006 to 2016 in the Plastic and Reconstructive Surgery Department of our institution. Only extensive facial defects greater than 40 cm2 were included in this study. RESULTS: A total of 323 patients underwent reconstruction using free flaps with or without local flaps, from November 2005 to March 2019. Of these, 79 extensive facial surface reconstruction cases were analyzed retrospectively. The size of the defects, their areas (upper, middle, or lower third of the face), and the method of reconstruction were analyzed. This led to the development of a personalized reconstruction procedure for extensive facial defects. These extensive facial reconstruction cases included 24 defects of the upper third of the face, 43 cases of the middle third, and 12 of the lower third. Four cases addressed the middle and lower thirds simultaneously. CONCLUSIONS: The facial surface reconstruction strategy I suggest in this study is based on the flap selection, thickness-controlled perforator flap elevation, and combination of local and free flaps. We expect this procedure will improve the treatment and resolution of extensive facial defects.

Structural and Electrical Properties of K(Ta, Nb)O3 Thin Films with the Variation of Sintering Temperature.

In this study, KTa0.55Nb0.45O3 (KTN) thin films were manufactured by using Chemical Solution Deposition (CSD) method with variations in the sintering temperature and were investigated in order to apply their applicability in memory devices. The KTN thin films were made after coating the PZT bufferlayer on Pt/Ti/SiO2/Si substrate. Each layer was dried at 200°C for 5 min to remove any organic materials and pyrolyzed at 400°C for 10 min. Finally, the layers were sintered for 30 min under an oxygen atmosphere, respectively. The pattern of KTN thin films showed a preference to the (100) and (200) orientations. Also, an increase in the sintering temperature caused the KTN crystalline peak intensities to also increase. When looking at the results from the Scanning Electron Microscope and Atomic Force Microscope data, the average grain size and root mean square roughness (Rrms) of KTN thin films were 109~157 nm and about 4 nm, respectively. Typical dielectric dispersion characteristics were observed in which the dielectric constant decreases with an increase of the applied frequency. The specimen sintered at 750°C showed the highest dielectric constant of 769 at 1 kHz.

Risk factor analysis of bone resorption following secondary alveolar bone grafting using three-dimensional computed tomography.

BACKGROUND/AIM: The purpose of this study is to analyze the risk factors for bone resorption following secondary bone grafting in the alveolar cleft, using three-dimensional (3D) computed tomography (CT) based on surgical simulation software (SimPlant OMS, Materialise Dental, Leuven, Belgium). METHODS: We reviewed the secondary alveolar bone grafts performed by a single surgeon between January 2005 and January 2014. A total of 40 patients with unilateral alveolar cleft were included in this study. The grafted alveolar bone was measured using surgical simulation software. In order to validate the measurement, each data set was measured by three different analysts and the inter- and intraobserver variabilities were calculated. A total of eight risk factors for grafted bone survival, including patient age, sex, body mass index (BMI), palatal fistula, amount of grafted bone, dental appliance, canine or incisor eruption, and preoperative upper respiratory tract infection, were evaluated using the linear mixed model and Mann-Whitney test. RESULTS: The average alveolar defect size was 4.98 cc and the average graft survival was 67.5%. The inter- and intraobserver variabilities of simulation software were 0.758 and 0.915, respectively. Among the risk factors, only dental appliance (p = 0.02) and canine eruption (p = 0.041) were significantly correlated with graft survival. Other risk factors, including the amount of grafted bone, did not show a significant relationship with graft survival. CONCLUSION: Measurement of an alveolar bone defect using a simulation program based on 3D CT is reliable and reproducible. Secondary bone grafting survival was significantly correlated with canine eruption and dental appliance in the alveolar cleft.

Lower lip deformity in patients with cleft and non-cleft Class III malocclusion before and after orthognathic surgery.

BACKGROUND: Orthognathic surgery does not yield the same cosmetic benefits in patients with Class III jaw deformities associated with clefts as for patients without clefts. Preoperative upper lip tightness caused by cleft lip repair may not fully explain this difference, suggesting that a lower lip deformity is present. The study compared the outcomes of orthognathic surgery in patients with cleft and non-cleft Class III malocclusion, focusing on lip relationship. MATERIALS AND METHODS: The surgical records of 50 patients with Class III malocclusion, including 25 with and 25 without clefts, who had undergone orthognathic surgery, were retrospectively analyzed. Lateral cephalometric tracings, preoperatively and at 6 months postoperatively, were superimposed to analyze the soft tissue changes at seven reference points. RESULTS: At 6 months after surgery, there were no significant differences in skeletal location, whereas the soft tissues of the lower lip differed significantly between patients with and without cleft (p=0.002), indicating the persistence of a lower lip deformity in cleft patients. Moreover, the soft tissues of the lower lip receded in non-cleft patients and protruded in cleft patients after orthognathic surgery. CONCLUSION: Lower lip deformity and upper lip tightness may result in an unsatisfactory relationship between the upper and lower lips of patients with cleft-related jaw deformity after orthognathic surgery. Other factors were less important than the pathology of the lower lip.

Using local flaps in a chest wall reconstruction after mastectomy for locally advanced breast cancer.

BACKGROUND: Surgical ablation for locally advanced breast cancer results in large chest wall defects, which can then be managed with local flaps or skin grafts. The purpose of this article is to evaluate the outcomes of three types of local skin flaps. METHODS: Among 25 local flaps in 24 patients, 6 were bilateral advancement (BA) flaps, 9 were thoracoabdominal (TA) flaps, and 10 were thoracoepigastric (TE) flaps. Clinical outcomes were compared including complications, the need for a secondary surgical intervention, and the timing of adjuvant therapy. RESULTS: The mean defect size was 436.2 cm(2). Two patients with TA flaps and 6 patients with TE flaps developed distal flap necrosis, and skin grafts were needed to treat 2 patients with TE flaps. Radiation was administered to the BA, TA, and TE patients after average postoperative durations of 28, 30, or 41 days, respectively. The incidence of flap necrosis tended to be higher in TE patients, which lead to significant delays in adjuvant radiation therapy (P=0.02). CONCLUSIONS: Three types of local skin flaps can be used to treat large chest wall defects after the excision of locally advanced breast cancer. Each flap has its own merits and demerits, and selecting flaps should be based on strict indications based on the dimensions and locations of the defects.

CSGM Designer: a platform for designing cross-species intron-spanning genic markers linked with genome information of legumes.

BACKGROUND: Genetic markers are tools that can facilitate molecular breeding, even in species lacking genomic resources. An important class of genetic markers is those based on orthologous genes, because they can guide hypotheses about conserved gene function, a situation that is well documented for a number of agronomic traits. For under-studied species a key bottleneck in gene-based marker development is the need to develop molecular tools (e.g., oligonucleotide primers) that reliably access genes with orthology to the genomes of well-characterized reference species. RESULTS: Here we report an efficient platform for the design of cross-species gene-derived markers in legumes. The automated platform, named CSGM Designer (URL: http://tgil.donga.ac.kr/CSGMdesigner), facilitates rapid and systematic design of cross-species genic markers. The underlying database is composed of genome data from five legume species whose genomes are substantially characterized. Use of CSGM is enhanced by graphical displays of query results, which we describe as "circular viewer" and "search-within-results" functions. CSGM provides a virtual PCR representation (eHT-PCR) that predicts the specificity of each primer pair simultaneously in multiple genomes. CSGM Designer output was experimentally validated for the amplification of orthologous genes using 16 genotypes representing 12 crop and model legume species, distributed among the galegoid and phaseoloid clades. Successful cross-species amplification was obtained for 85.3% of PCR primer combinations. CONCLUSION: CSGM Designer spans the divide between well-characterized crop and model legume species and their less well-characterized relatives. The outcome is PCR primers that target highly conserved genes for polymorphism discovery, enabling functional inferences and ultimately facilitating trait-associated molecular breeding.

Use of a serratus anterior musculocutaneous flap for surgical obliteration of a bronchopleural fistula.

OBJECTIVES: Successful management of post-resection bronchopleural fistula (BPF) is a challenge, and various loco-regional flaps have been used to directly cover or to enhance closure of the bronchial stump. The serratus anterior muscle (SAM) is one of the workhorse flaps, although its use as a musculocutaneous flap has been debated. Here, we present our early experience with serratus anterior musculocutaneous (SAMC) flaps for the obliteration of BPF. METHODS: A retrospective review of patients with surgical management of BPF from April 2005 to June 2014 was performed. A de-epithelized SAMC flap has replaced the conventional SAM flap since August 2013. Nine of consecutive former SAM flaps and 5 consecutive later SAMC flaps were identified. A detailed review of the SAMC flap cases was performed to describe medical and surgical history, BPF diagnosis and location, and the outcome of the flap surgery. RESULTS: All five BPFs treated by SAMC flap were greater than 1 cm in diameter and all occurred on the right side. The leading primary diagnosis of a BPF was lung cancer, and the 4 lung cancer patients all underwent previous irradiation. The average size of the skin paddle was 19 × 6.6 cm(2), and the average volume of the skin paddle was 100.3 cm(3). Recurrence of the BPF was noted in 1 patient 6 weeks after surgery. In 4 of 5 patients, viable adipose tissue was confirmed by computed tomography scans. CONCLUSIONS: The SAMC flap holds merits of a local flap such as short operation time, relative safety, no need of position change and faster recovery, with increased freedom of flap transfer and additional volume, thus widening surgical indication of the large, recalcitrant BPFs. We suggest that the SAMC flap could be safely harvested and used for management of a BPF or similar intrathoracic lesion when extensive flap length or bulk is required.