Comparison of the Pharmacokinetics of Gadolinium-Based and Iron Oxide-Based Contrast Agents inside the Lymphatic Structure using Magnetic Resonance Lymphangiography.

PURPOSE: Gadolinium (Gd)-based contrast agents are primarily used for contrast-enhanced magnetic resonance lymphangiography (MRL). However, overcoming venous contamination issues remains challenging. This study aims to assess the MRL efficacy of the newly developed iron-based contrast agent (INV-001) that is specially designed to mitigate venous contamination issues. The study further explores the optimal dosage, including both injection volume and concentration, required to achieve successful visualization of the popliteal lymph nodes and surrounding lymphatic vessels. PROCEDURES: All animals utilized in this study were male Sprague-Dawley (SD) rats weighing between 250 and 300 g. The contrast agents prepared were injected intradermally in the fourth phalanx of both hind limbs using a 30-gauge syringe in SD rats. MRL was performed every 16 min on a coronal 3D time-of-flight sequence with saturation bands using a 9.4-T animal machine. RESULTS: Contrary to Gd-DOTA, which exhibited venous contamination in most animals irrespective of injection dosages and conditions, INV-001 showed no venous contamination. For Gd-DOTA, the popliteal lymph nodes and lymphatic vessels reached peak enhancement 16 min after injection from the injection site and then rapidly washed out. However, with INV-001, they reached peak enhancement between 16 and 32 min after injection, with prolonged visualization of the popliteal lymph node and lymphatic vessels. INV-001 at 0.45 µmol (15 mM, 30 µL) and 0.75 µmol (15 mM, 50 µL) achieved high scores for qualitative image analysis, providing good visualization of the popliteal lymph nodes and lymphatic vessels without issues of venous contamination, interstitial space enhancement, or lymph node enlargement. CONCLUSION: In MRL, INV-001, a novel T1 contrast agent based on iron, enables prolonged enhancement of popliteal lymph nodes and lymphatic vessels without venous contamination.

Margin of exposure to free formaldehyde in personal care products containing formaldehyde-donor preservatives: Evidence for consumer safety.

Formaldehyde has been classified as carcinogenic to humans by International Agency for Research on Cancer and found in personal care (PC) products containing formaldehyde-donor (FD) preservatives. However, the cancer risk associated with the use of FD-containing PC products has not been well established. Our study provides the quantitative cancer risk assessment of formaldehyde in FD-containing PC products. The carbon-13 nuclear magnetic resonance (13C-NMR) spectroscopy was used in this risk assessment to provide reliable exposure information to formaldehyde in PC products and aqueous solutions containing sodium hydroxymethylglycinate. The risk assessment was conducted using the margin of exposure (MOE) approach with benchmark doses (BMDs) for 10% effect. For hemolymphoreticular neoplasias in male rats, a BMD of 28.03 mg/kg/day and a BMD lower confidence limit (BMDL) of 2.52 mg/kg/day were calculated from available long-term animal experiments. The worst-case consumer exposure to formaldehyde from FD-containing PC products was 0.007 µg/kg/day. Comparing the consumer exposure with BMDL, the resulting MOE was 360,000 for the worst-case scenario. The consumer exposure to formaldehyde (0.007 µg/kg/day) from using FD-containing PC products represents less than 1.0 × 10-6 % of background level endogenous formaldehyde (878-1310 mg/kg/day). The cancer risk from formaldehyde to consumers using FD-containing PC products is negligible.

MicroRNA super-resolution imaging in blood for Alzheimer's disease.

We propose a novel blood biomarker detection method that uses miRNA super-resolution imaging to enable the early diagnosis of Alzheimer's disease (AD). Here, we report a singlemolecule detection method for visualizing disease-specific miRNA in tissue from an AD mice model, and peripheral blood mononuclear cells (PBMCs) from AD patients. Using optimized Magnified Analysis of Proteome (MAPs), we confirmed that five miRNAs contribute to neurodegenerative disease in the brain hippocampi of 5XFAD and wild-type mice. We also assessed PBMCs isolated from the whole blood of AD patients and a healthy control group, and subsequently analyzed those samples using miRNA super-resolution imaging. We detected more miR-200a-3p expression in the cornu ammonis 1 and dentate gyrus regions of 3 month-old 5XFAD mice than in wild-type mice. Additionally, miRNA super-resolution imaging of blood provides AD diagnosis platform for studying miRNA regulation inside cells at the single molecule level. Our results present a potential liquid biopsy method that could improve the diagnosis of early stage AD and other diseases. [BMB Reports 2023; 56(3): 190-195].

Genotyping-by-sequencing-based identification of Arabidopsis pattern recognition receptor RLP32 recognizing proteobacterial translation initiation factor IF1.

Activation of plant pattern-triggered immunity (PTI) relies on the recognition of microbe-derived structures, termed patterns, through plant-encoded surface-resident pattern recognition receptors (PRRs). We show that proteobacterial translation initiation factor 1 (IF1) triggers PTI in Arabidopsis thaliana and related Brassicaceae species. Unlike for most other immunogenic patterns, IF1 elicitor activity cannot be assigned to a small peptide epitope, suggesting that tertiary fold features are required for IF1 receptor activation. We have deployed natural variation in IF1 sensitivity to identify Arabidopsis leucine-rich repeat (LRR) receptor-like protein 32 (RLP32) as IF1 receptor using a restriction site-associated DNA sequencing approach. RLP32 confers IF1 sensitivity to rlp32 mutants, IF1-insensitive Arabidopsis accessions and IF1-insensitive Nicotiana benthamiana, binds IF1 specifically and forms complexes with LRR receptor kinases SOBIR1 and BAK1 to mediate signaling. Similar to other PRRs, RLP32 confers resistance to Pseudomonas syringae, highlighting an unexpectedly complex array of bacterial pattern sensors within a single plant species.

Treatment of chemotherapy-induced cachexia with BST204: a multimodal validation study.

INTRODUCTION: Chemotherapy is a major etiology of cachexia. Ginseng products are known to have various anti-cachectic and health-promoting effects, such as inhibiting inflammation and promoting energy production. In particular, BST204, purified ginseng dry extract, contains multiple ginsenosides that can reduce chemotherapy-related fatigue and toxicity. OBJECTIVES: To investigate the effects of BST204 on the alleviation of chemotherapy-induced cachexia using a multimodal approach. METHODS: In a CT26 mouse syngeneic colon cancer model, cachexia was predominantly induced by chemotherapy with 5-fluorouracil (5-FU) than by tumor growth. BST204 at a dose of 100 or 200 mg/kg was administered to 5-FU-treated mice. RESULTS: BST204 significantly mitigated the decrease in tumor-excluded body weight (change in 5-FU group and BST204 groups: - 13% vs. - 6% on day 7; - 30% vs. - 20% on day 11), muscle volume (- 19% vs. - 11%), and fat volume (- 91% vs. - 56%). The anti-cachectic effect of BST204 was histologically demonstrated by an improved balance between muscle regeneration and degeneration and a decrease in muscle cross-sectional area reduction. CONCLUSION: Chemotherapy-induced cachexia was biochemically and metabolically characterized by activated inflammation, enhanced oxidative stress, increased protein degradation, decreased protein stabilization, reduced glucose-mediated energy production, and deactivated glucose-mediated biosynthesis. These adverse effects were significantly improved by BST204 treatment. Overall, our multimodal study demonstrated that BST204 could effectively alleviate chemotherapy-induced cachexia.

Off-the-Shelf, Immune-Compatible Human Embryonic Stem Cells Generated Via CRISPR-Mediated Genome Editing.

Human embryonic stem cells (hESCs) hold promise in regenerative medicine but allogeneic immune rejections caused by highly polymorphic human leukocyte antigens (HLAs) remain a barrier to their clinical applications. Here, we used a CRISPR/Cas9-mediated HLA-editing strategy to generate a variety of HLA homozygous-like hESC lines from pre-established hESC lines. We edited four pre-established HLA-heterozygous hESC lines and created a mini library of 14 HLA-edited hESC lines in which single HLA-A and HLA-B alleles and both HLA-DR alleles are disrupted. The HLA-edited hESC derivatives elicited both low T cell- and low NK cell-mediated immune responses. Our library would cover about 40% of the Asian-Pacific population. We estimate that HLA-editing of only 19 pre-established hESC lines would give rise to 46 different hESC lines to cover 90% of the Asian-Pacific population. This study offers an opportunity to generate an off-the-shelf HLA-compatible hESC bank, available for immune-compatible cell transplantation, without embryo destruction. Graphical Abstract.

Anesthetic management of obese and morbidly obese parturients.

The prevalence of obese parturients is increasing worldwide. This review describes safe analgesic techniques for labor and anesthetic management during cesarean sections in obese parturients. The epidural analgesic technique is the best way to provide good pain relief during the labor phase and can be easily converted to a surgical anesthetic condition. However, the insertion of the epidural catheter in obese parturients is technically more difficult compared to that in non-obese parturients. The distance from the skin to the epidural space increases in proportion to the body mass index (BMI): 4.4 cm in mothers of normal weight and 7.5 cm in mothers with BMI 50 and above. Neuraxial blocks are the ideal anesthetic methods and gold standard techniques for cesarean section in pregnant women with obesity. Single-shot spinal anesthesia is the most common type of anesthesia used for cesarean sections. The advantage of single-shot spinal anesthesia is a dense-sufficient block of rapid onset. A combined spinal-epidural (CSE) anesthetic technique is also recommended as an attractive alternative method. In obese parturients, the operation time can be longer than expected, and therefore, the CSE technique provides the advantage of rapid onset and intense block for prolonged operation with postoperative pain control. The risk of postoperative complications is very high in obese parturients. Therefore, detailed communication of the parturient's medical condition and the details of surgery and anesthesia between the anesthesiologist and obstetrician is important prior to cesarean section in obese pregnant women.

Non-coplanar VMAT plans for lung SABR to reduce dose to the heart: a planning study.

OBJECTIVE: This study aimed to compare the plan quality of non-coplanar partial arc (NPA) volumetric modulated arc therapy (VMAT) to that of coplanar partial arc (CPA) VMAT for stereotactic ablative radiotherapy (SABR) for lung cancer. METHODS: A total of 20 patients treated for lung cancer with the SABR VMAT technique and whose lung tumors were close to the heart were retrospectively selected for this study. For the CPA VMAT, three coplanar half arcs were used while two coplanar half arcs and one noncoplanar arc rotating 315°-45° with couch rotations of 315° ± 5° were used for the NPA VMAT. For each patient, identical CT image sets and identical structures were used for both the CPA and NPA VMAT plans. Dose-volumetric parameters of each plan were analyzed. RESULTS: For the planning target volume and both lungs, no statistically significant differences between the CPA and NPA VMAT plans were observed in general. For the heart, average values of D0.1cc of the CPA and NPA VMAT plans were 29.42 ± 13.37 and 21.71 ± 9.20 Gy, respectively (p < 0.001). For whole body, the mean dose and the gradient index of the CPA VMAT plans were 1.2 ± 0.5 Gy and 4.356 ± 0.608 while those of the NPA VMAT plans were 1.1 ± 0.5 Gy and 4.111 ± 0.480, respectively (both with p < 0.001). CONCLUSION: The NPA VMAT proposed in this study showed more favorable plan quality than the CPA VMAT plans for lung SABR with tumors located close to the heart. ADVANCES IN KNOWLEDGE: For lung SABR, NPA VMAT can reduce doses to the heart as well as whole-body irradiation.

Modeling scattered radiation from multi-leaf collimators (MLCs) to improve calculation accuracy of in-air output ratio.

This study aims to model an extra-focal source for the scattered radiation from multi-leaf collimators (MLCs), namely an MLC scatter source, and to correct in-air output ratio (Sc) calculated using the conventional dual source model (DSM) to achieve better accuracy of point dose calculation. To develop the MLC scatter source, a 6 MV photon beam from a Varian Clinac® iX linear accelerator with millennium 120 MLCs was used. It was assumed that the position for the MLC scatter source was located at the center of the MLC, consisting of line-based and area-based sources to consider the characteristics of the scattered radiation from the MLCs empirically. Based on the measured Sc values for MLC-defined fields, optimal parameters for the line-based and area-based sources were calculated using optimization process. For evaluation of proposed method, measurements were taken for various MLC-defined square and irregular fields. The Sc values calculated using the proposed MLC scatter source and conventional DSM were compared with the measured data. For MLC-defined square fields, the measured Sc values showed better agreement with those calculated using the MLC scatter source (the mean difference was - 0.03%) compared with those calculated using the DSM (the mean difference was 0.18%). For MLC-defined irregular fields, the maximum dose differences between measurements and calculations using the MLC scatter source and DSM were 0.54% and 1.45%, respectively. The developed MLC scatter source could improve the accuracy of Sc calculation for both square and irregular fields defined by MLCs.

Magnetic Resonance Colonography Enables the Efficacy Assessment of Immune Checkpoint Inhibitors in an Orthotopic Colorectal Cancer Mouse Model.

Immune checkpoint inhibitors (ICIs) have become an effective therapeutic option for colorectal cancer and studies on these drugs have therefore increased greatly. Efficacy assessments of ICIs in preclinical orthotopic colorectal cancer using MRI have not been reported however due to the difficulties in conducting colorectal imaging. The purpose of this present study was to investigate the feasibility of using magnetic resonance colonography (MRC) to evaluate the efficacy of an ICI, an anti-PD-L1 antibody, in an orthotopic colorectal cancer mouse model. The mouse model was generated by the engraftment of colorectal cancer cells into the submucosal layer of the colon. Anti-cancer efficacy was assessed by tumor volume and metastatic tumor number analyses, and these values were significantly lower in the PD-L1 antibody-treated group compared to the controls. Histological analyses using H&E and Ki-67 immunohistochemical staining confirmed a highly efficacious tumor growth inhibition and enhanced infiltration by CD4+ and CD8+ lymphocytes in the PD-L1 antibody-treated group. We conclude that MRC has the potential to be used for ICI efficacy assessments against orthotopic colorectal cancer mouse model.

ANALYSIS OF THE RADIOLOGICAL SAFETY CONTROL LEVEL VERSUS THE EXPOSURE OF RADIATION WORKERS IN SOUTH KOREA FROM 2008-17.

In this study, the radiation exposure of workers at workplaces registered and licensed between 2008 and 2017 for the production/sale/use of radioactive isotopes (RI) and radioactive generators (RG) was analysed to evaluate the quality of radiation safety management controls in use. The number of facilities using RIs increased by ~26% from 2008 to 2017 whereas the number of facilities using RGs increased by ~166% over the same period. There were 33 029 radiation workers in all fields in 2008, and the number increased by ~32% to 43 467 by 2017. However, the collective effective dose of radiation received by workers decreased in all industries except for those working in nuclear power plants. In other words, the quality of radiation safety management improved over that same time period due to the systematic, continuous introduction of safety mechanisms by the regulatory authority.

Ultrasensitive Fluorescence Monitoring and in Vivo Live Imaging of Circulating Tumor Cell-Derived miRNAs Using Molecular Beacon System.

Circulating tumor cells (CTCs) have considerable clinical significance in cancer progression and prognosis. In this context, CTC-derived microRNAs (miRs) in blood and tissues have been proposed as the novel noninvasive biomarkers for monitoring tumor progression, especially at the early stages. To monitor circulating miRs, a tool should have high sensitivity, be a simple procedure, and allow detection in very small volumes. Thus, we designed a sensing tool for sensitive monitoring of blood or tissue miRs using a fluorophore-quencher probe-based molecular beacon (MB). This MB-based tool displayed an ultrasensitive limit of detection (LOD) level of 6.7 × 10-17 M and 8.7 × 10-17 for metastasis-derived miR-21a and miR-221, respectively. It also can discriminate miR-21a/221 from both guide strand miRs and its precursor forms (pre-miR). Furthermore, the tool discriminated between blood and tissue-related miR-21a/221-expression and detected metastasis and epithelial-mesenchymal transition and also describe a noninvasive miR fluorescence imaging of CTCs in a mouse model, showing the potential for clinical diagnosis and prognosis.

Reliability of the gamma index analysis as a verification method of volumetric modulated arc therapy plans.

BACKGROUND: We investigate the gamma passing rate (GPR) consistency when applying different types of gamma analyses, linacs, and dosimeters for volumetric modulated arc therapy (VMAT). METHODS: A total of 240 VMAT plans for various treatment sites, which were generated with Trilogy (140 plans) and TrueBeam STx (100 plans), were retrospectively selected. For each VMAT plan, planar dose distributions were measured with both MapCHECK2 and ArcCHECK dosimeters. During the planar dose distribution measurements, the actual multileaf collimator (MLC) positions, gantry angles, and delivered monitor units were recorded and compared to the values in the original VMAT plans to calculate mechanical errors. For each VMAT plan, both the global and local gamma analyses were performed with 3%/3 mm, 2%/2 mm, 2%/1 mm, 1%/2 mm, and 1%/1 mm. The Pearson correlation coefficients (r) were calculated 1) between the global and the local GPRs, 2) between GPRs with the MapCHECK2 and the ArcCHECK dosimeters, 3) and between GPRs and the mechanical errors during the VMAT delivery. RESULTS: For the MapCHECK2 measurements, strong correlations between the global and local GPRs were observed only with 1%/2 mm and 1%/1 mm (r > 0.8 with p < 0.001), while weak or no correlations were observed for the ArcCHECK measurement. Between the MapCHECK2 and ArcCHECK measurements, the global GPRs showed no correlations (all with p > 0.05), while the local GPRs showed moderate correlations only with 2%/1 mm and 1%/1 mm for TrueBeam STx (r > 0.5 with p < 0.001). Both the global and local GPRs always showed weak or no correlations with the MLC positional errors except for the GPRs of MapCHECK2 with 1%/2 mm and 1%/1 mm for TrueBeam STx and the GPR of ArcCHECK with 1%/2 mm for Trilogy (r < - 0.5 with p < 0.001). CONCLUSIONS: The GPRs varied according to the types of gamma analyses, dosimeters, and linacs. Therefore, each institution should carefully establish their own gamma analysis protocol by determining the type of gamma index analysis and the gamma criterion with their own linac and their own dosimeter.

Adenine base editing in mouse embryos and an adult mouse model of Duchenne muscular dystrophy.

Adenine base editors (ABEs) composed of an engineered adenine deaminase and the Streptococcus pyogenes Cas9 nickase enable adenine-to-guanine (A-to-G) single-nucleotide substitutions in a guide RNA (gRNA)-dependent manner. Here we demonstrate application of this technology in mouse embryos and adult mice. We also show that long gRNAs enable adenine editing at positions one or two bases upstream of the window that is accessible with standard single guide RNAs (sgRNAs). We introduced the Himalayan point mutation in the Tyr gene by microinjecting ABE mRNA and an extended gRNA into mouse embryos, obtaining Tyr mutant mice with an albino phenotype. Furthermore, we delivered the split ABE gene, using trans-splicing adeno-associated viral vectors, to muscle cells in a mouse model of Duchenne muscular dystrophy to correct a nonsense mutation in the Dmd gene, demonstrating the therapeutic potential of base editing in adult animals.

The impact of acute inflammation on progression and metastasis in pancreatic cancer animal model.

BACKGROUND: The impact of acute inflammation on cancer is unclear. Therefore, we evaluated the impact of acute inflammation on cancer progression in an animal model concerning morphological change as well as molecular features. MATERIAL AND METHODS: Murine pancreas ductal adenocarcinoma cell line (Panc-02) and zymosan were used for the induction of cancer and acute inflammation respectively, in C57/BL6 mice. In the C group (n = 10), 2 × 107Panc02 cells were injected into the tail of the pancreas. In the Z1 (n = 10) and Z2 (n = 10), 3 mg of zymosan was injected intraperitoneally once and twice respectively, after tumor cell injection. All of the mice were sacrificed 4 weeks after tumor cell injection. RESULTS: The degree of inflammation was more severe in the Z2 group than in the other two groups. The tumor volume of the Z2 group was larger than that of the C group (P = 0.032) and the presence of liver metastasis was significantly more common in the Z2 group than in the C and Z1 groups (P = 0.025). Confocal microscopy analysis revealed significantly more expression of Snail, EpCAM, Muc1, NLRP3 and miR-155 in the liver and pancreas in the Z2 group than in the other two groups. EpCAM and Muc1 were detected in blood samples in the Z2 group by flow cytometry, but not in the other two groups. In the Z2, expression of E-Cadherin was weaker and vimentin, snail1 were stronger compared to the C and Z1 group in the PCR and western blot. CONCLUSION: Present results suggest that acute inflammation accompanied with cancer promotes cancer progression by the epithelial-to-mesenchymal transition (EMT) and circulating tumor cells (CTC) process.

Intravenously Administered Lidocaine and Magnesium During Thyroid Surgery in Female Patients for Better Quality of Recovery After Anesthesia.

BACKGROUND: Although systemic lidocaine and magnesium have been widely studied as perioperative analgesic adjuvants, they have been rarely evaluated with respect to recovery quality under the same conditions. We compared the quality of recovery 40 (QoR-40) scores of female patients who received intravenous lidocaine, magnesium, and saline during thyroidectomy to investigate their effects on comprehensive recovery from anesthesia. METHODS: In this prospective, double-blind trial, 135 female patients scheduled for open thyroidectomy were randomly assigned to the lidocaine group (group L), magnesium group (group M), or control group (group C). Immediately after induction, lidocaine (2 mg/kg for 15 minutes followed by 2 mg/kg/h) was administered in group L and magnesium sulfate (20 mg/kg over 15 minutes followed by 20 mg/kg/h) was administered in group M. Group C received an equivalent volume of saline. The QoR-40 survey was conducted on postoperative days 1 and 2. RESULTS: The mean global QoR-40 scores on postoperative day 1 were 186.3 (standard deviation, 5.5) in group L, 184.3 (4.7) in group M, and 179.4 (17.8) in group C, and there was a significant difference only between group L and group C (mean difference, 6.9; adjusted P = .018). Among the 5 dimensions of QoR-40, emotional state, physical comfort, and pain were superior in group L compared to group C. CONCLUSIONS: Lidocaine administered intravenously during anesthesia led to better quality of postoperative recovery measured by QoR-40 compared with the group C. Magnesium was found to be insufficient to induce any significant improvement with the dose used in the present study.

On-site audits to investigate the quality of radiation physics of radiation therapy institutions in the Republic of Korea.

PURPOSE: To investigate and improve the domestic standard of radiation therapy in the Republic of Korea. METHODS: On-site audits were performed for 13 institutions in the Republic of Korea. Six items were investigated by on-site visits of each radiation therapy institution, including collimator, gantry, and couch rotation isocenter check; coincidence between light and radiation fields; photon beam flatness and symmetry; electron beam flatness and symmetry; physical wedge transmission factors; and photon beam and electron beam outputs. RESULTS: The average deviations of mechanical collimator, gantry, and couch rotation isocenter were less than 1mm. Those of radiation isocenter were also less than 1mm. The average difference between light and radiation fields was 0.9±0.6mm for the field size of 20cm×20cm. The average values of flatness and symmetry of the photon beams were 2.9%±0.6% and 1.1%±0.7%, respectively. Those of electron beams were 2.5%±0.7% and 0.6%±1.0%, respectively. Every institutions showed wedge transmission factor deviations less than 2% except one institution. The output deviations of both photon and electron beams were less than ±3% for every institution. CONCLUSIONS: Through the on-site audit program, we could effectively detect an inappropriately operating linacs and provide some recommendations. The standard of radiation therapy in Korea is expected to improve through such on-site audits.

Genome-wide target specificities of CRISPR RNA-guided programmable deaminases.

Cas9-linked deaminases, also called base editors, enable targeted mutation of single nucleotides in eukaryotic genomes. However, their off-target activity is largely unknown. Here we modify digested-genome sequencing (Digenome-seq) to assess the specificity of a programmable deaminase composed of a Cas9 nickase (nCas9) and the deaminase APOBEC1 in the human genome. Genomic DNA is treated with the base editor and a mixture of DNA-modifying enzymes in vitro to produce DNA double-strand breaks (DSBs) at uracil-containing sites. Off-target sites are then computationally identified from whole genome sequencing data. Testing seven different single guide RNAs (sgRNAs), we find that the rAPOBEC1-nCas9 base editor is highly specific, inducing cytosine-to-uracil conversions at only 18 ± 9 sites in the human genome for each sgRNA. Digenome-seq is sensitive enough to capture off-target sites with a substitution frequency of 0.1%. Notably, off-target sites of the base editors are often different from those of Cas9 alone, calling for independent assessment of their genome-wide specificities.

Usefulness of Dermoscopy in the Differential Diagnosis of Ruptured and Unruptured Epidermal Cysts.

BACKGROUND: An epidermal cyst is a common keratin-filled epithelial-lined cyst. The treatment of choice for epidermal cysts is surgical excision. If the cyst becomes ruptured, incision and drainage with oral antibiotic therapy or intralesional steroid injection are required. OBJECTIVE: To analyze the dermoscopic features that can differentiate between ruptured and unruptured epidermal cysts. METHODS: The clinical and dermoscopic features of the pathologically confirmed epidermal cysts of two subgroups of 38 patients, 20 with unruptured cysts and 18 with ruptured cysts, were reviewed. RESULTS: With regard to the dermoscopic features, an ivory- white background color and punctum were commonly found in both groups (p>0.05). The unruptured-cyst group showed higher frequencies of pore sign (p<0.05), blue-white veil (p>0.05), no vascular structure, and arborizing telangiectasia (p<0.05), but the ruptured-cyst group usually had red lacunae (p>0.05) and peripheral linear branched vessels (with an erythematous rim) (p<0.05). CONCLUSION: Dermoscopy is helpful in differentiating between ruptured and unruptured epidermal cysts.

CRISPR/Cpf1-mediated DNA-free plant genome editing.

Cpf1, a type V CRISPR effector, recognizes a thymidine-rich protospacer-adjacent motif and induces cohesive double-stranded breaks at the target site guided by a single CRISPR RNA (crRNA). Here we show that Cpf1 can be used as a tool for DNA-free editing of plant genomes. We describe the delivery of recombinant Cpf1 proteins with in vitro transcribed or chemically synthesized target-specific crRNAs into protoplasts isolated from soybean and wild tobacco. Designed crRNAs are unique and do not have similar sequences (≤3 mismatches) in the entire soybean reference genome. Targeted deep sequencing analyses show that mutations are successfully induced in FAD2 paralogues in soybean and AOC in wild tobacco. Unlike SpCas9, Cpf1 mainly induces various nucleotide deletions at target sites. No significant mutations are detected at potential off-target sites in the soybean genome. These results demonstrate that Cpf1-crRNA complex is an effective DNA-free genome-editing tool for plant genome editing.