An efficient method for constructing a random insertional mutant library for forward genetics in Nannochloropsis oceanica

Insertional mutation,phenotypic evaluation,and mutated gene cloning are widely used to clone genes from scratch.Exogenous genes can be integrated into the genome during non-homologous end joining(NHEJ)of the double-strand breaks of DNA,causing insertional mutation.The random insertional mutant library constructed using this method has become a method of forward genetics for gene cloning.However,the establishment of a random insertional mutant library requires a high transformation efficiency of exogenous genes.Many microalgal species show a low transformation efficiency,making constructing random insertional mutant libraries difficult.In this study,we established a highly efficient transformation method for constructing a random insertional mutant library of Nannochloropsis oceanica,and tentatively tried to isolate its genes to prove the feasibility of the method.A gene that may control the growth rate and cell size was identified.This method will facilitate the genetic studies of N.oceanica,which should also be a reference for other microalgal species.

Efficient isolation of specific genomic regions by insertional chromatin immunoprecipitation (iChIP) with a second-generation tagged LexA DNA-binding domain

Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. We have developed a novel method, insertional chromatin immunoprecipitatin (iChIP), to isolate specific genomic regions retaining molecular interaction in order to perform non-biased identification of interacting molecules in vivo. Here, we developed a second-generation tagged LexA DNA-binding domain, 3xFNLDD, for the iChIP analysis. 3xFNLDD consists of 3 x FLAG tags, a nuclear localization signal (NLS), the DNA-binding domain (DB) and the dimerization domain of the LexA protein. Expression of 3xFNLDD can be detected by immunoblot analysis as well as flowcytometry. We showed that iChIP using 3xFNLDD is able to consistently isolate more than 10% of input genomic DNA, several-fold more efficient compared to the first-generation tagged LexA DB. 3xFNLDD would be a useful tool to perform the iChIP analysis for locus-specific biochemical epigenetics.

Ultrasound and Color Doppler-Guided Surgery for Insertional Achilles Tendinopathy-Results of a Pilot Study

Background: Treatment of insertional achilles tendinopathy is known to be difficult. Eccentric calf muscle exercises and extracorporeal shockwave therapy have been proposed as the primary treatments for this condition, but surgery is indicated after failed conservative management. There is no consensus about the most efficient surgical treatment. Objectives: To evaluate a new ultrasound and color Doppler-guided surgical treatment for insertional achilles tendinopathy. Patients and methods: 24 consecutive patients (13 men and 11 women, mean age 47 years) with a long duration of insertional achilles tendon pain (median 18 months) were included in the study. The surgical procedure consisted of extirpation of the subcutaneous and retrocalcaneal bursa, scraping of the ventral distal achilles tendon and removal of prominent bone at the upper calcaneal tuberosity (Haglund’s deformity). In 13 patients, the plantaris tendon was also cut and excised. VAS for pain during activity, satisfaction with the treatment result and the SF-36 score to measure quality of life were used for evaluation. Results: The mean VAS had decreased from 72 before surgery to 19 after surgery (p 0.001), and 20/24 (83%) patients were satisfied with the result and had returned to full tendon loading activity at the one-year follow-up. The SF-36 score had improved significantly. Conclusion: Ultrasound and color Doppler-guided surgical treatment seems to be a good method for treatment of chronic painful insertional achilles tendinopathy. Longer follow-up studies on larger materials are needed.

Isolating the Mutator Transposable Element Insertional Mutant Gene mio16 of Maize Using Double SelectedAmplification of Insertion Flanking Fragments (DSAIFF)

Mutator transposable element （Mu） has been used as an effective tool to clone maize （Zea mays L.） genes. One opaque endosperm mutant （miol6） was identified in a pool of Mu inserted mutants. A modified method, termed the double selected amplification of insertion flanking fragments （DSAIFF）, was employed to isolate the Mu flanking fragments （MFFs） of miol6. The target site duplications （TSDs） isolated from the Msp I and Mse I digested MFFs had a same 9-bp sequence and were confirmed to be the flanking sequence of one identically inserted gene. Co-segregation analysis suggested that the MFFs were associated with the mutant opaque endosperm, and miol6 was mapped in silico onto the physical position ranged from 229 965 021 to 229 965 409 bp of the maize chromosome 4.09 bin. The full-length cDNA of the wild-type gene was obtained by an RT-PCR primer-scanning technique, and Mio16 was found to putatively encode a homolog of the Arabidopsis MAP3K delta-1 protein kinase. RT-PCR result the mRNA expression of miol6 region anchored by primers Mu20 and af276 was not interrupted by Mu insertion. Further researches will be done to elucidate how the expression of miol6 is alternated by Mu insertion.

Esophageal inserted subclavian catheter:a rare complication

Central venous catheterization(CVC) is an invasive procedure for administering fluids,nutrients,and drugs;monitoring central venous pressure;performing pulmonary artery catheterization;and placing transvenous pacemakers in intensive care units and all specialties,from anesthesia to emergency medicine,for the treatment of trauma and hemodynamically unstable pediatric and adult patients.[1,2]Complications have been observed in more than 15% of patients who underwent CVC.Mechanical,infectious,and thrombotic complications have been reported in 5%–19%,5%–26%,and 2%–26% of patients,respectively.[3] Malposition,on the other hand,is common in subclavian catheter insertion and is usually associated with an initially misplaced guidewire.[4]

Reversible Optical Isolators and Quasi-Circulators Using a Magneto-Optical Fabry-Pérot Cavity

Nonreciprocal optical devices are essential for laser protection,modern optical communication and quantum information processing by enforcing one-way light propagation.The conventional Faraday magneto-optical nonreciprocal devices rely on a strong magnetic field,which is provided by a permanent magnet.As a result,the isolation direction of such devices is fixed and severely restricts their applications in quantum networks.In this work,we experimentally demonstrate the simultaneous one-way transmission and unidirectional reflection by using a magneto-optical Fabry-Pérot cavity and a magnetic field strength of 50 mT.An optical isolator and a three-port quasi-circulator are realized based on this nonreciprocal cavity system.The isolator achieves an isolation ratio of up to 22 dB and an averaged insertion loss down to 0.97 dB.The quasi-circulator is realized with a fidelity exceeding 99% and an overall survival probability of 89.9%,corresponding to an insertion loss of~0.46 dB.The magnetic field is provided by an electromagnetic coil,thereby allowing for reversing the light circulating path.The reversible quasi-circulator paves the way for building reconfigurable quantum networks.

Lower extremity peripherally inserted central catheter placement ectopic to the ascending lumbar vein:A case report

BACKGROUND Peripherally inserted central catheters(PICCs)are an essential infusion route for oncology patients receiving intravenous treatments,but lower extremity veni-puncture is the preferred technique for patients with superior vena cava syndrome(SVCS).We report the case of a patient with a lower extremity PICC ectopic to the ascending lumbar vein,to indicate and verify PICC catheterisation in the lower extremity is safe and feasible.And hope to provide different per-spectives for clinical PICC venipuncture to get the attention of peers.CASE SUMMARY On 24 August 2022,a 58-year-old male was admitted to our department due to an intermittent cough persisting for over a month,which worsened 10 d prior.Imaging and laboratory investigations suggested the patient with pulmonary malignancy and SVCS.Chemotherapy was not an absolute contraindication in this patient.Lower extremity venipuncture is the preferred technique because administering upper extremity venous transfusion to patients with SVCS can exacerbate oedema in the head,neck,and upper extremities.The patient and his family were informed about the procedure,and informed consent was obtained.After successful puncture and prompt treatment,the patient was discharged,experiencing some relief from symptoms.CONCLUSION Inferior vena cava catheterisation is rare and important for cancer patients with SVCS,particularly in complex situations involving ectopic placement.

Tubular Heat Enhancement Using Twisted Tape Inserts with Large Holes

Heat augmentation techniques play a vital role in the heating and cooling processes in industries,including solar collectors and many applications that utilize heat exchangers.Several studies are based on inserting fillers inside the tubes to enhance heat transfer.This investigation considered the effects of twisted tapes with large holes on a tubular heat exchanger’s(HX)heat transmission,pressure drop,and thermal boosting factor.In the experimental section,counter-swirl flow generators used twisted tapes with pairs of 1.0 cm-diameter holes and changes in porosity(Rp)at 1.30%and 2.70%.In the experiments,air was utilized as a working fluid in a tube with a circular cross-section.The turbulent flow was considered,with Reynolds numbers(Re)domain from 4800 to 9500,and a boundary condition with a uniform wall heat flux was constructed.The findings expound that when the number of holes rose,the Nusselt number(Nu),the factor of friction(f),and the thermal enhancement factor(η)all increased as well.Additionally,as the friction factor increased,the Nusselt number of the tape-equipped tube was noticeably higher.Additionally,it was discovered that the friction factor was between 70%and 94%lower than the values of the tube without tape,while the(Nu)was between 87%and 97%higher than the conventional tube values.The maximum value ofηis 89%.According to the experimental results,empirical correlations for Nu,f,andηwere also generated.

Treatment of benign rectal stricture caused by repeated anal insertion by endoscopy and balloon dilation:A case report

BACKGROUND Benign rectal strictures can be categorized as primary(disease-related)and secondary(surgical anastomosis-related).Secondary strictures arise from surgical complications,whereas primary strictures have diverse etiologies,including various inflammatory conditions.Benign strictures are usually managed by surgery and endoscopy.We present an unusual etiology of benign rectal stricture caused by the repeated insertion of foreign objects into the rectum for sexual purposes,resulting in rectal injury and subsequent chronic inflammation.CASE SUMMARY A 53-year-old man presented to the outpatient clinic of the Colorectal Surgery Department with symptoms of chronic constipation and bloody stools.The patient previously experienced rectal injury due to foreign object insertion for sexual purposes.Colonoscopy revealed benign circumferential narrowing of the rectum.He underwent treatment by endoscopic argon plasma coagulation and balloon dilation and follow-up as an outpatient for 4 months.A colonoscopy at the end of the follow-up period revealed no evidence of rectal stricture relapse.CONCLUSION A history of rectal injury,followed by chronic inflammation,should be considered in patients with benign rectal strictures.Management with endoscopic argon plasma coagulation and balloon dilation can prevent the need for surgical resection of benign rectal strictures.

Investigation of Fracture Strength of Current Post Materials (Peek, Fiber, Cast Metal) in Different Ferrule Conditions

Aim: The fracture resistance of the post materials used nowadays significantly impacts the fracture type of the post material. The current study aimed to examine the impact of the fracture resistance of PEEK posts, whose use as a post material has not yet become widespread, on the fracture pattern that will occur. Materials and Methods: Sixty-six extracted upper central incisor human teeth were used in the study. Six groups of posts were produced from metal, fiber, and PEEK materials (n = 11). Afterward, the fracture resistance test was applied to the samples. Samples were classified according to fracture type as repairable or irreparable. The ANOVA and test were used in statistical analysis. Post-hoc tests were carried out using Tukey’s and Tamhane’s T2 tests. On the other hand, Pearson’s chi-square test and the Fisher-Freeman-Halton test were conducted for comparisons between categorical variables. The statistical significance level was accepted as p Results: According to the fracture resistance test results, the groups where metal personal posts were used displayed higher fracture resistance in comparison with all other groups. According to the results of the optical microscope examination, the incidence of repairable fractures was significantly higher in the PEEK post group with ferrule preparation (p 0.05). Conclusion: In the absence of the ferrule effect, fractures are catastrophic, regardless of the material used. The use of PEEK as a post material contributes to keeping fractures at a repairable level.

Natural Gas Power Plants in the System Security Role—The Situation and Solutions in Brazil

This study aims to apply a methodology to analyze the regulatory obstacles for the development of thermal power plants in a country. The study uses Brazil as an application case, although mostly of the findings can be useful to other countries. It also discusses other policies and actions by the government that may contribute to the growth of natural gas-fired power plants. Using deductive and comparative research methods, the first part of this method focuses on the stage of thermoelectric power generation in Brazil. The subsequent sections present the role of the integration of gas and electricity sectors, the main obstacles to the expansion of natural-gas power generation, how the country has deal with the expansion of natural-gas power generation, as well as how other obstacles has been faced. In raising the research question, when comparing the research results of selected countries, it should be noticed the impact that the experience and lessons elsewhere have had on policies formulation on Brazil. Our conclusions pointed out that adequate policies for the natural gas-electricity Brazilian market integration make advisable the unification of electricity and natural gas agencies. We recommended developing a mathematical model to support decision-making on natural gas and electricity integration.

Current Topics in Acute Abdomen: Causes, Diagnostics, and Emerging Therapies

Acute abdomen is a serious clinical condition that requires immediate attention due to its association with intense pain and rapid progression. This article reviews important aspects of the causes, diagnostics, and treatments of acute abdomen. The research highlights the diversity of etiologies, ranging from the most common, such as acute appendicitis, to the less frequent, such as intestinal perforations by foreign bodies. Complex diagnostic methods, including clinical examinations and imaging such as computed tomography, are essential to guide accurate diagnosis, given the overlap of symptoms among different conditions. Therapeutic approaches range from conservative treatments with clinical observation and antibiotic therapy to urgent surgical interventions, such as appendectomy and cholecystectomy. Laparoscopy stands out as a promising approach, offering advantages in terms of recovery and post-operative morbidity. However, the impact of the COVID-19 pandemic has brought additional challenges to the diagnosis and treatment of acute abdomen, with delays and difficulties in accessing healthcare. This review underscores the importance of a multidisciplinary approach and constant updates among healthcare professionals in addressing acute abdomen, aiming for accurate diagnoses and appropriate therapeutic interventions to improve clinical outcomes and reduce morbidity and mortality associated with this complex clinical condition.

Velamentous Cord Insertion: Interest of Antenatal Diagnosis and Review of the Literature

Velamentous insertion of the umbilical cord corresponds to the insertion of the cord directly on amniotic membranes. It is a rare situation whose frequency varies from 0.5% to 1.69% of single pregnancies. It must be diagnosed during the morphological ultrasound of the 2nd trimester, actively looking for the association with a vasa previa, due to the risk of fetal haemorrhagic threat. We report an antenatal diagnosis of velamentous cord insertion and its management with literature review.

耳内窥镜下鼓膜置管联合腺样体切除术与鼓膜穿刺治疗分泌性中耳炎患儿的临床疗效比较

目的探讨耳内窥镜下鼓膜置管联合腺样体切除术治疗分泌性中耳炎(OME)患儿的临床疗效。方法选取2021年1月~2022年3月SOM患儿120例,按术式分为穿刺组(n=60)和置管组(n=60),穿刺组给予耳内窥镜下鼓膜穿刺联合腺样体切除术,置管组给予耳内窥镜下鼓膜置管联合腺样体切除术,比较两组积液消退时间、听觉脑干诱发电位(ABR)阈值、ABRⅠ波潜伏期、干扰素γ(IFN-γ)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、治疗效果以及并发症。结果置管组积液消退时间显著低于穿刺组,差异有统计学意义(P<0.05);穿刺组和置管组术后ABR、ABRⅠ波潜伏期、IFN-γ、IL-6和TNF-α均显著低于术前(P均<0.05);置管组术后ABR、ABRⅠ波潜伏期、IFN-γ、IL-6和TNF-α均显著低于穿刺组,差异有统计学意义(P均<0.05);置管组治疗有效率显著高于穿刺组,差异有统计学意义(P<0.05);置管组并发症发生率显著低于穿刺组,差异有统计学意义(P<0.05)。结论耳内窥镜下鼓膜置管联合腺样体切除术可有效改善SOM患儿听力及炎症,有利于提高疗效,且可减少并发症,值得临床推广。

Stem cell gene therapy:the risks of insertional mutagenesis and approaches to minimize genotoxicity

Virus-based vectors are widely used in hematopoietic stem cell(HSC)gene therapy,and have the ability to integrate permanently into genomic DNA,thus driving long-term expression of corrective genes in all hematopoietic lineages.To date,HSC gene therapy has been successfully employed in the clinic for improving clinical outcomes in small numbers of patients with X-linked severe combined immunodeficiency(SCID-X1),adenosine deaminase deficiency(ADA-SCID),adrenoleukodystrophy(ALD),thalassemia,chronic granulomatous disease(CGD),and Wiskott-Aldrich syndrome(WAS).However,adverse events were observed during some of these HSC gene therapy clinical trials,linked to insertional activation of proto-oncogenes by integrated proviral vectors leading to clonal expansion and eventual development of leukemia.Numerous studies have been performed to understand the molecular basis of vector-mediated genotoxicity,with the aim of developing safer vectors and lower-risk gene therapy protocols.This review will summarize current information on the mechanisms of insertional mutagenesis in hematopoietic stem and progenitor cells due to integrating gene transfer vectors,discuss the available assays for predicting genotoxicity and mapping vector integration sites,and introduce newlydeveloped approaches for minimizing genotoxicity as a way to further move HSC gene therapy forward into broader clinical application.

TEAseq-based identification of 35,696 Dissociation insertional mutations facilitates functional genomic studies in maize

In plants,transposable element(TE)-triggered mutants are important resources for functional genomic studies.However,conventional approaches for genome-wide identification of TE insertion sites are costly and laborious.This study developed a novel,rapid,and high-throughput TE insertion site identification workflow based on next-generation sequencing and named it Transposable Element Amplicon Sequencing(TEAseq).Using TEAseq,we systemically profiled the Dissociation(Ds)insertion sites in 1606 independent Ds insertional mutants in advanced backcross generation using K17 as background.The Ac-containing individuals were excluded for getting rid of the potential somatic insertions.We characterized 35,696 germinal Ds insertions tagging 10,323 genes,representing approximately 23.3%of the total genes in the maize genome.The insertion sites were presented in chromosomal hotspots around the ancestral Ds loci,and insertions occurred preferentially in gene body regions.Furthermore,we mapped a loss-of-function AGL2 gene using bulked segregant RNA-sequencing assay and proved that AGL2 is essential for seed development.We additionally established an open-access database named MEILAM for easy access to Ds insertional mutations.Overall,our results have provided an efficient workflow for TE insertion identification and rich sequence-indexed mutant resources for maize functional genomic studies.

Relationships between distribution characteristics of ceramic fragments and anti-penetration performance of ceramic composite bulletproof insert plates

Through quantitative statistics and morphological characterization of ceramic fragments for ceramic composite bulletproof insert plates(CCBIPs),distribution characteristics of ceramic fragments within a specific size range were analyzed for different Armor Piercing Incendiary(API)and shot times.To quantitatively evaluate the effect of energy absorption for ceramic plates,a model of energy absorption during penetration for CCBIPs was established based on statistics of the size distribution of ceramic fragments(SDCF).Variation in the SDCF and its influence on energy absorption for CCBIPs were investigated.The results indicate that the distribution feature of ceramic fragments in the range of 0.25-2.25 mm is Gaussian distribution.Compared with Type 56 of API(56-API),ceramic fragments formed by 53-API with higher kinetic energy possess more quantity and more concentrated distribution,whose average equivalence size decreases by 6.5%,corresponding to increasing by 83.9%of estimated energy absorption.Besides,the ability of CCBIPs to resist the third shot is significantly weakened,whose estimated energy absorption decreases by 58.8%compared with the first shot.More concentrated distribution and fewer fragments are formed after the third shot,the average equivalence size of ceramic fragments increases by 6.9%,which may attribute to the micro-cracks induced by the previous two shots.

Efficient large fragment deletion in plants:double pairs of sgRNAs are better than dual sgRNAs

Dear Editor,Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)is a powerful and versatile gene editing system that has been extensively utilized in various animals and plants,which holds enormous potential and value for scientific research and breeding.However,single-targeted CRISPR can only induce a few base deletions,insertions,or substitution.Ideally,thesemutations result in premature termination of the protein encoded by the target gene,leading to a loss of function[1].

Health Monitoring of Milling Tool Inserts Using CNN Architectures Trained by Vibration Spectrograms

In-process damage to a cutting tool degrades the surface􀀀nish of the job shaped by machining and causes a signi􀀀cant􀀀nancial loss.This stimulates the need for Tool Condition Monitoring(TCM)to assist detection of failure before it extends to the worse phase.Machine Learning(ML)based TCM has been extensively explored in the last decade.However,most of the research is now directed toward Deep Learning(DL).The“Deep”formulation,hierarchical compositionality,distributed representation and end-to-end learning of Neural Nets need to be explored to create a generalized TCM framework to perform eciently in a high-noise environment of cross-domain machining.With this motivation,the design of dierent CNN(Convolutional Neural Network)architectures such as AlexNet,ResNet-50,LeNet-5,and VGG-16 is presented in this paper.Real-time spindle vibrations corresponding to healthy and various faulty con􀀀gurations of milling cutter were acquired.This data was transformed into the time-frequency domain and further processed by proposed architectures in graphical form,i.e.,spectrogram.The model is trained,tested,and validated considering dierent datasets and showcased promising results.

Atomic-scale insights of indium segregation and its suppression by GaAs insertion layer in InGaAs/AlGaAs multiple quantum wells

The In segregation and its suppression in InGaAs/AlGaAs quantum well are investigated by using high-resolution x-ray diffraction(XRD)and photoluminescence(PL),combined with the state-of-the-art aberration corrected scanning transmission electron microscopy(Cs-STEM)techniques.To facility our study,we grow two multiple quantum wells(MQWs)samples,which are almost identical except that in sample B a thin GaAs layer is inserted in each of the InGaAs well and AlGaAs barrier layer comparing to pristine InGaAs/AlGaAs MQWs(sample A).Our study indeed shows the direct evidences that In segregation occurs in the InGaAs/AlGaAs interface,and the effect of the Ga As insertion layer on suppressing the segregation of In atoms is also demonstrated on the atomic-scale.Therefore,the atomic-scale insights are provided to understand the segregation behavior of In atoms and to unravel the underlying mechanism of the effect of GaAs insertion layer on the improvement of crystallinity,interface roughness,and further an enhanced optical performance of InGaAs/AlGaAs QWs.