Efficient Screening Method Research Articles

Monitoring of Water Spectral Pattern Reveals Differences in Probiotics Growth When Used for Rapid Bacteria Selection.

Development of efficient screening method coupled with cell functionality evaluation is highly needed in contemporary microbiology. The presented novel concept and fast non-destructive method brings in to play the water spectral pattern of the solution as a molecular fingerprint of the cell culture system. To elucidate the concept, NIR spectroscopy with Aquaphotomics were applied to monitor the growth of sixteen Lactobacillus bulgaricus one Lactobacillus pentosus and one Lactobacillus gasseri bacteria strains. Their growth rate, maximal optical density, low pH and bile tolerances were measured and further used as a reference data for analysis of the simultaneously acquired spectral data. The acquired spectral data in the region of 1100-1850nm was subjected to various multivariate data analyses – PCA, OPLS-DA, PLSR. The results showed high accuracy of bacteria strains classification according to their probiotic strength. Most informative spectral fingerprints covered the first overtone of water, emphasizing the relation of water molecular system to cell functionality.

Stochastic Curtailment of Questionnaires for Three-Level Classification: Shortening the CES-D for Assessing Low, Moderate, and High Risk of Depression.

In clinical assessment, efficient screeners are needed to ensure low respondent burden. In this article, Stochastic Curtailment (SC), a method for efficient computerized testing for classification into two classes for observable outcomes, was extended to three classes. In a post hoc simulation study using the item scores on the Center for Epidemiologic Studies-Depression Scale (CES-D) of a large sample, three versions of SC, SC via Empirical Proportions (SC-EP), SC via Simple Ordinal Regression (SC-SOR), and SC via Multiple Ordinal Regression (SC-MOR) were compared at both respondent burden and classification accuracy. All methods were applied under the regular item order of the CES-D and under an ordering that was optimal in terms of the predictive power of the items. Under the regular item ordering, the three methods were equally accurate, but SC-SOR and SC-MOR needed less items. Under the optimal ordering, additional gains in efficiency were found, but SC-MOR suffered from capitalization on chance substantially. It was concluded that SC-SOR is an efficient and accurate method for clinical screening. Strengths and weaknesses of the methods are discussed.

Production of monoclonal antibodies against GPCR using cell-free synthesized GPCR antigen and biotinylated liposome-based interaction assay.

G-protein-coupled receptors (GPCRs) are one of the most important drug targets, and anti-GPCR monoclonal antibody (mAb) is an essential tool for functional analysis of GPCRs. However, it is very difficult to develop GPCR-specific mAbs due to difficulties in production of recombinant GPCR antigens, and lack of efficient mAb screening method. Here we describe a novel approach for the production of mAbs against GPCR using two original methods, bilayer-dialysis method and biotinylated liposome-based interaction assay (BiLIA), both of which are developed using wheat cell-free protein synthesis system and liposome technology. Using bilayer-dialysis method, various GPCRs were successfully synthesized with quality and quantity sufficient for immunization. For selection of specific mAb, we designed BiLIA that detects interaction between antibody and membrane protein on liposome. BiLIA prevented denaturation of GPCR, and then preferably selected conformation-sensitive antibodies. Using this approach, we successfully obtained mAbs against DRD1, GHSR, PTGER1 and T1R1. With respect to DRD1 mAb, 36 mouse mAbs and 6 rabbit mAbs were obtained which specifically recognized native DRD1 with high affinity. Among them, half of the mAbs were conformation-sensitive mAb, and two mAbs recognized extracellular loop 2 of DRD1. These results indicated that this approach is useful for GPCR mAb production.

Development of an Efficient Method of Screening for Watermelon Plants Resistant to Fusarium oxysporum f. sp. niveum

This study was conducted to establish an efficient screening method for watermelon plants resistant to Fusarium wilt (FW), which is caused by Fusarium oxysporum f. sp. niveum (Fon). An HA isolate was prepared from a wilted watermelon plant in Haman-gun and identified as F. oxysporum f. sp. niveum based on morphological characteristics, molecular analyses of ITS (internal transcribed spacer) and TEF (translation elongation factor ) sequences, and host specificity on cucurbits including watermelon, melon, oriental melon, and cucumber. The assay for disease response of watermelon differentials indicated that the HA isolate was race 0. Among seven liquid media tested, the highest amount of Fon spores was produced from V8-juice broth, which was selected as a medium for mass production of Fon. The disease assay for 21 watermelon and 11 watermelon-rootstock cultivars demonstrated that 20 watermelon cultivars except for 'Soknoranggul' were susceptible; 'Soknoranggul' was moderately resistant. All the tested rootstock cultivars were highly resistant to the HA isolate. The evaluation of disease development depending on various conditions suggested that an efficient screening method for FW resistance in watermelon plants is to dip the roots of 10-day-old seedlings in spore suspension of for 30 min., to transplant the seedlings to plastic pots with a fertilized soil, and then to cultivate the plants at for 3 weeks.

Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes.

BackgroundClustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated genome editing permits the rapid production of genetically engineered mice. To make the most of this innovative technology, a streamlined procedure is needed for the robust construction of CRISPR/Cas9 vectors, the efficient preparation of mouse oocytes, and refined genotyping strategies. Although we previously demonstrated the applicability of oocyte cryopreservation technologies and various genotyping methods in the production of transcription activator-like effector nuclease-mediated genome editing in mice, it has not yet been clarified whether these techniques can be applied to the CRISPR/Cas9-mediated generation of knockout mice. In this study, we investigated easy, efficient, and robust methods of creating knockout mice using several CRISPR/Cas9 systems.ResultsWe constructed three types of CRISPR/Cas9 vectors expressing: 1) single guide RNA (gRNA) and Cas9 nuclease, 2) two gRNAs and Cas9 nickase, and 3) two gRNAs and FokI-dCas9, targeting the same genomic locus. These vectors were directly microinjected into the pronucleus of freeze-thawed fertilized oocytes, and surviving oocytes were transferred to pseudopregnant ICR mice. Cas9 nuclease resulted in the highest mutation rates with the lowest birth rates, while Cas9 nickase resulted in the highest birth rates with the lowest mutation rates. FokI-dCas9 presented well-balanced mutation and birth rates. Furthermore, we constructed a single all-in-one FokI-dCas9 vector targeting two different genomic loci, and validated its efficacy by blastocyst analysis, resulting in highly efficient simultaneous targeted mutagenesis.ConclusionsOur report offers several choices of researcher-friendly consolidated procedures for making CRISPR/Cas9-mediated knockout mice, with sophisticated construction systems for various types of CRISPR vectors, convenient preparation of in vitro fertilized or mated freeze-thawed oocytes, and an efficient method of mutant screening.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-015-0144-x) contains supplementary material, which is available to authorized users.

Comparison of Electronic Screening for Suicidal Risk With the Patient Health Questionnaire Item 9 and the Columbia Suicide Severity Rating Scale in an Outpatient Psychiatric Clinic

Patient-reported data can improve clinical care, including identifying patients who are at risk for suicide. In a tertiary care, psychiatric outpatient clinic, we compared computerized self-assessments of suicidal risk based on item 9 of the Patient Health Questionnaire-9 and an electronic version of the Columbia Suicide Severity Rating Scale (C-SSRS), using retrospective medical record review of clinical psychiatric assessments as the reference standard. We also surveyed patients׳ attitudes about participating in the process. We compared prevalence of suicidal risk rates by the 3 assessment methods as well as their sensitivity, specificity, and predictive value. Observed prevalence of positive suicidal risk screenings differed significantly, ranking (1) Patient Health Questionnaire-9 item 9, 24% (343/1416; 95% CI: 22%-26%) < (2) C-SSRS, 6.0% (85/1416; 95% CI: 5.0%-7.4%) < (3) clinical assessment, 1.4% (20/1416; 95% CI: 0.9%-2.2%). The sensitivity of Patient Health Questionnaire-9 item 9 was 92% (78/85; 95% CI: 86%-98%) and the specificity was 81% (1107/1376; 95% CI: 78%-82%). The sensitivity of the C-SSRS was 95.0% (19/20; 95% CI: 75%-100%) and the specificity was 95% (1330/1396; 95% CI: 94%-96%). Of 100 patients surveyed, the screening was well accepted, with some concerns about confidentiality and adequate clinical follow-up. As expected, Patient Health Questionnaire-9 item 9 generated much higher rates of apparently false-positive findings than the C-SSRS did, when compared with clinical assessment. C-SSRS backed with timely clinical assessment may be a useful and efficient method of screening for suicidal risk, provided that adequate, immediate clinical follow-up is available.

Screening for lung cancer using low-dose computed tomography: concerns about the application in low-risk individuals.

Low-dose computed tomography (LDCT) has been increasingly accepted as an efficient screening method for high-risk individuals to reduce lung cancer mortality. However, there remains a gap of knowledge in the practical implementation of screening on a larger scale, especially for low-risk individuals. The aim of this study is to initiate discussion through an evidence-based analysis and provide valuable suggestions on LDCT screening for lung cancer in clinical practice. Among previously published randomized controlled trials (RCTs), the National Lung Screening Trial (NLST) is the only one demonstrating positive results in a high-risk population of old age and heavy smokers. It is also shown that the potential harms include false-positive findings, radiation exposure etc., but its magnitude is uncertain. In the meantime, the current risk stratification system is inadequate, and is difficult to define selection criteria. Thus, the efficacy of LDCT in lung cancer screening needs to be confirmed in future trials, and the procedure should not be proposed to individuals without comparable risk to those in the NLST. Furthermore, there is a lack of evidence to support the expansion of LDCT screening to low-risk individuals. Therefore, recommendation of LDCT screening for these patients could be premature in clinical practice although some of them might be missed based on current definition of risk factors. Further studies and advances in risk assessment tools are urgently needed to address the concerns about lung cancer screening in order to improve the outcomes of lung cancer.

An efficient screening method for simultaneous detection of recurrent copy number variants associated with psychiatric disorders

Several recurrent copy number variants (CNVs) increasing risk to neuropsychiatric diseases have been identified in recent years. They show variable clinical expressivity, being associated with different disorders, and incomplete penetrance. However, due to its very low frequency, the full variety of clinical outcomes associated with each one of these CNVs is unknown. Current methods for detection of CNVs are labor intensive, expensive or not suitable for high throughput analysis. Quantitative interspecies competitive PCR linked to variant minisequencing and detection by mass-spectrometry may overcome these limitations. Here, we present two multiplex assays based on this method to screen for eleven psychiatric risk CNVs, such as 1q21, 16p11.2, 3q29, or 16p13.11 regions, among others. The assays were tested in our collection of 514 schizophrenia patients. Results were compared with MLPA at two CNVs. Additional positive results were confirmed by exome sequencing. A total of fourteen patients were CNV carriers. The method presents high sensitivity and specificity, showing its utility as a cheap, accurate, high throughput screening tool for recurrent CNVs. The method may be very useful for management of psychiatric patients as well as screening of different collections of samples to better identify the full spectrum of clinical variability.

Radionuclide, scintillation cocktail and chemical/color quench influence on discriminator setting in gross alpha/beta measurements by LSC

Gross alpha/beta measurements in drinking waters enable radiochemical composition analysis in environmental studies providing efficient screening method that can indicate whether water contains elevated levels of any radionuclide. Routine gross alpha/beta activity monitoring in drinking waters has been carried out for a few years in laboratory for low-level radioactivity measurements in Novi Sad according to ASTM method, performing measurements on liquid scintillation counter Quantulus 1220 which can simultaneously generate alpha/beta spectra of samples by Pulse Shape Analysis (PSA circuit). In this paper, PSA discriminator impact was investigated to ensure obtaining of accurate and reliable alpha/beta activities. One novelty of presented work is PSA parameter setup with two combinations of radionuclides (241Am, 226Ra and 90Sr/90Y) with varying activity concentrations. Performed experiments also make contribution to investigations on the manner in which chemical and color quench affect optimal PSA parameter setting and further on, their altogether influence on gross alpha/beta activity measurements. Nitromethane, 15.8 M nitric acid and water, as well as yellow and yellow-orange dye, were used as quenching agents in order to test PSA/interference factor behavior in the presence of quenchers with different quenching strengths. Variation of PSA setting in quenched samples with two different commercially available cocktails (Ultima Gold LLT and OptiPhase HiSafe 3) was also tested. Lastly, application i.e. assessment of obtained PSA-SQP(E) correlation on the obtained results of activity concentrations of few artesian well water samples and colored spiked samples, based on the measured SQP(E) value of samples, has been demonstrated.

An Efficient Screening Method for Detection of Toluene-based Inhalant Abuse in Urine by Gas Chromatography-Nitrogen Phosphorous Detector

An Efficient Screening Method for Detection of Toluene-based Inhalant Abuse in Urine by Gas Chromatography-Nitrogen Phosphorous Detector

Virtual screening using combinatorial cyclic peptide libraries reveals protein interfaces readily targetable by cyclic peptides.

Protein-protein and protein-peptide interactions are responsible for the vast majority of biological functions in vivo, but targeting these interactions with small molecules has historically been difficult. What is required are efficient combined computational and experimental screening methods to choose among a number of potential protein interfaces worthy of targeting lead macrocyclic compounds for further investigation. To achieve this, we have generated combinatorial 3D virtual libraries of short disulfide-bonded peptides and compared them to pharmacophore models of important protein-protein and protein-peptide structures, including short linear motifs (SLiMs), protein-binding peptides, and turn structures at protein-protein interfaces, built from 3D models available in the Protein Data Bank. We prepared a total of 372 reference pharmacophores, which were matched against 108,659 multiconformer cyclic peptides. After normalization to exclude nonspecific cyclic peptides, the top hits notably are enriched for mimetics of turn structures, including a turn at the interaction surface of human α thrombin, and also feature several protein-binding peptides. The top cyclic peptide hits also cover the critical "hot spot" interaction sites predicted from the interaction crystal structure. We have validated our method by testing cyclic peptides predicted to inhibit thrombin, a key protein in the blood coagulation pathway of important therapeutic interest, identifying a cyclic peptide inhibitor with lead-like activity. We conclude that protein interfaces most readily targetable by cyclic peptides and related macrocyclic drugs may be identified computationally among a set of candidate interfaces, accelerating the choice of interfaces against which lead compounds may be screened.

Childhood Trauma Exposure and Toxic Stress: What the PNP Needs to Know

Trauma exposure in childhood is a major public health problem that can result in lifelong mental and physical health consequences. Pediatric nurse practitioners must improve their skills in the identification of trauma exposure in children and their interventions with these children. This continuing education article will describe childhood trauma exposure (adverse childhood experiences) and toxic stress and their effects on the developing brain and body. Adverse childhood experiences include a unique set of trauma exposures. The adverse childhood experiences or trauma discussed in this continuing education offering will include childhood exposure to emotional abuse, physical abuse, sexual abuse, emotional neglect, physical neglect, domestic violence, household substance abuse, household mental illness, parental separation or divorce, and a criminal household member. Thorough and efficient methods of screening for trauma exposure will be discussed. Appropriate intervention after identification of trauma exposure will be explored.

Development of Efficient Screening Methods for Melon Plants Resistant to Fusarium oxysporum f. sp. melonis

본 연구는 덩굴쪼김병균(Fusarium oxysporum f. sp. melonis)에 대한 저항성 멜론의 효율적인 검정법을 확립하기 위하여 수행하였다. 고령에서 채집한 덩굴쪼김병이 발생한 멜론으로부터 GR 균주를 분리하였으며, 형태학적 및 분자생물학적 동정 방법에 의해 그리고 오이, 멜론, 참외, 수박의 박과 작물에 대한 기주 특이성 조사를 통하여 GR 균주는F. oxysporum f. sp. melonis로 동정되었다. 그리고 4종 덩굴 쪼김병 race 판별품종들의 저항성 반응에 따라 GR 균주는 race 1임을 알 수 있었다. GR 균주의 접종원(소형분생포자) 대량생산을 위해서는 실험한 6종 액체배지 중 V8-juice broth에서 가장 많은 포자가 형성되었다. 시판 중인 22개의 멜론 품종과 6개 멜론 재배용 대목 품종의 GR 균주에 대한 저항성의 정도를 실험하였다. 실험한 멜론 품종 중 3개 품종을 제외한 모든 품종은 다양한 정도의 저항성을 보였다. 그리고 실험한 대목 품종들 모두에서는 덩굴쪼김병이 전혀 발생하지 않았다. 실험한 멜론 품종 중 GR 균주에 대한 저항성 반응에 차이를 보이는 6개 품종('레드퀸', '썸머쿨', '슈퍼세지', '아시아파파야', '얼룩파파야', '아시아황금')을 선발하여 멜론 생육시기, 뿌리 상처, 침지 시간, 접종원 농도 및 재배 온도 등의 발병 조건에 따른 덩굴쪼김병 발생을 조사하였다. 이들 실험의 결과로부터 멜론 품종들의 덩굴쪼김병에 대한 저항성을 검정하기 위해서는 멜론 종자를 파종하고 온실(<TEX>$25{\pm}5^{\circ}C$</TEX>)에서 7일 동안 재배한 유묘(떡잎 시기)를 뽑아 흙을 제거하고 뿌리 자르기와 같은 상처를 내지 않고 멜론 유묘의 뿌리를 <TEX>$3{\times}10^5conidia/mL$</TEX> 농도의 F. oxysporum f. sp. melonis 포자현탁액에 30분 정도 침지하여 접종하고, 이를 새로운 토양에 이식하고 <TEX>$25-28^{\circ}C$</TEX>에서 약 3주일 동안 재배하는 것이 가장 효율적인 방법임을 알 수 있었다. This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of <TEX>$3{\times}10^5conidia/mL$</TEX> for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to <TEX>$28^{\circ}C$</TEX> for about 3 weeks with 12-hour light per day.

Live fluorescent RNA-based detection of pluripotency gene expression in embryonic and induced pluripotent stem cells of different species.

The generation of induced pluripotent stem (iPS) cells has successfully been achieved in many species. However, the identification of truly reprogrammed iPS cells still remains laborious and the detection of pluripotency markers requires fixation of cells in most cases. Here, we report an approach with nanoparticles carrying Cy3-labeled sense oligonucleotide reporter strands coupled to gold-particles. These molecules are directly added to cultured cells without any manipulation and gene expression is evaluated microscopically after overnight incubation. To simultaneously detect gene expression in different species, probe sequences were chosen according to interspecies homology. With a common target-specific probe we could successfully demonstrate expression of the GAPDH house-keeping gene in somatic cells and expression of the pluripotency markers NANOG and GDF3 in embryonic stem cells and iPS cells of murine, human, and porcine origin. The population of target gene positive cells could be purified by fluorescence-activated cell sorting. After lentiviral transduction of murine tail-tip fibroblasts Nanog-specific probes identified truly reprogrammed murine iPS cells in situ during development based on their Cy3-fluorescence. The intensity of Nanog-specific fluorescence correlated positively with an increased capacity of individual clones to differentiate into cells of all three germ layers. Our approach offers a universal tool to detect intracellular gene expression directly in live cells of any desired origin without the need for manipulation, thus allowing conservation of the genetic background of the target cell. Furthermore, it represents an easy, scalable method for efficient screening of pluripotency which is highly desirable during high-throughput cell reprogramming and after genomic editing of pluripotent stem cells.

An efficient blue-white screening based gene inactivation system for Streptomyces.

Streptomyces is studied intensively for its outstanding ability to produce bioactive secondary metabolites and for its complicated morphological differentiation process. A classical genetic manipulation system for Streptomyces has been developed and widely used in the community for a long time, using antibiotic resistance markers to select for double-crossover mutants. The screening process is always laborious and time-consuming. However, the lack of a suitable chromogenic reporter for Streptomyces has limited the use of color-based screening system to simplify the selection process for double-crossover mutants. In this study, a blue reporter system for Streptomyces has been established by mining an indigoidine synthetase gene (idgS) from Streptomyces lavendulae CGMCC 4.1386, leading to the development of a time-saving gene inactivation system for Streptomyces by simple blue-white screening. A series of Streptomyces suicide and temperature-sensitive plasmids containing the idgS reporter cassette were constructed and used successfully to inactivate genes in Streptomyces, allowing a simple and efficient screening method to differentiate the colonies for double-crossover (white) and single-crossover (blue) mutants. Inactivation of the putative γ-butyrolactone synthase gene afsA-y via the idgS-based blue-white screening method revealed that the paulomycin production is negatively controlled by afsA-y in Streptomyces sp. YN86.

New approach for natural products screening by real-time monitoring of hemoglobin hydrolysis using quartz crystal microbalance

The hemoglobin (Hb) released from erythrocytes is a primary nutritive component for many blood-feeding parasites. The aspartic protease cathepsin D is a hemoglobinase that is involved in the Hb degradation process and is considered an interesting target for chemotherapy intervention. However, traditional enzymatic assays for studying Hb degradation utilize spectrophotometric techniques, which do not allow real-time monitoring and can present serious interference problems. Herein, we describe a biosensor using simple approach for the real-time monitoring of Hb hydrolysis as well as an efficient screening method for natural products as enzymatic inhibitors using a quartz crystal microbalance (QCM) technique. Hemoglobin was anchored on the quartz crystal surface using mixed self-assembled monolayers. The addition of the enzyme caused a mass change (frequency shift) due to Hb hydrolysis, which was monitored in real time. From the frequency change patterns of the Hb-functionalized QCM, we evaluated the enzymatic reaction by determining the kinetic parameters of product formation (kcat). The QCM enzymatic assay using immobilized human Hb was shown to be an excellent approach for screening possible inhibitors in complex mixtures, opening up a new avenue for the discovery of novel inhibitors.

Efficient screening of 2D molecular polymorphs at the solution-solid interface.

Formation of multiple polymorphs during two-dimensional (2D) crystallization of organic molecules is more of a routine occurrence than rarity. Although such diverse crystalline structures provide exciting possibilities for studying crystal engineering in 2D, predicting the occurrence of polymorphs for a given building block is often non-trivial. Moreover, there is scarcity of methods that can experimentally verify the presence of such crystalline polymorphs in a straightforward fashion. Here we demonstrate a relatively simple experimental approach for screening of 2D polymorphs formed at the solution-solid interface. The strategy involves use of solution flow produced by contacting a piece of tissue paper to the sample to generate a lateral density gradient along the substrate surface. In situ generation of such gradient allows rapid discovery and nanoscale separation of multiple 2D polymorphs in a single experiment. The concept is demonstrated using three structurally different building blocks that differ in terms of intermolecular interactions responsible for 2D crystal formation. The method described here represents a powerful tool for efficient screening of 2D polymorphs formed at the solution-solid interface.

Challenges in Sonographic Detection of Fetal Major Structural Abnormalities at the First Trimester Anomaly Scan

ABSTRACT First-trimester (FT) ultrasound examination appears to have a very good sensitivity in the detection of fetal abnormalities, especially when an extended protocol that is achievable with reasonable resources of time, personnel and ultrasound equipment, is used. It has been shown that the efficiency of the routine anomaly scan varies widely between the studies performed both in early or mid-pregnancy. The analysis of technique and study designs reveals that in each gestational age, the extension of the investigational protocol is the most important factor in modulating the detection rate. Moreover, the use of similar basic checklists at the FT and second trimester (ST) examinations revealed comparable anomaly detection rates in large population groups. First trimester combined test have been demonstrated as a useful screening tool for genetic syndromes and consecutively the FT ultrasound scan became a routine examination in most prenatal diagnostic centers. Its performance in structural abnormalities detection has been communicated in high-, medium-risk populations or unselected low-risk variable number of patients and compared to the ST anomaly scan effectiveness. A detailed first-trimester anomaly scan using an extended protocol is an efficient screening method to detect major fetal structural abnormalities in low-risk pregnancies. It is a method that it is feasible at 12 to 13 + 6 weeks with ultrasound equipment and personnel already used for routine first-trimester screening. Rate of detection of severe malformations is greater in early- than in mid-pregnancy and on postnatal evaluation. Early heart investigation could be improved by an extended protocol involving use of color Doppler. How to cite this article Iliescu D, Cara M, Tudorache S, Antsaklis P, Ceausu I, Paulescu D, Novac L, Cernea N, Antsaklis A. Challenges in Sonographic Detection of Fetal Major Structural Abnormalities at the First Trimester Anomaly Scan. Donald School J Ultrasound Obstet Gynecol 2015;9(3):239-259.

Mannosylerythritol lipids: production and applications.

Mannosylerythritol lipids (MELs) are a glycolipid class of biosurfactants produced by a variety yeast and fungal strains that exhibit excellent interfacial and biochemical properties. MEL-producing fungi were identified using an efficient screening method for the glycolipid production and taxonomical classification on the basis of ribosomal RNA sequences. MEL production is limited primarily to the genus Pseudozyma, with significant variability among the MEL structures produced by each species. Outside of Pseudozyma, one recently isolated strain, Ustilago scitaminea, has been shown to exhibit abundant MEL-B production from sugarcane juice. Structural analyses of these compounds suggest a role for MELs in numerous cosmetic applications. MELs act as effective topical moisturizers and can repair damaged hair. Furthermore, these compounds have been shown to exhibit both protective and healing activities, to activate fibroblasts and papilla cells, and to act as natural antioxidants. In this review, we provide a brief summary of MEL research over the past few decades, focusing on the identification of MEL-producing fungi, the structural characterization of MELs, the use of alternative compounds as a primary carbon source, and the use of these compounds in cosmetic applications.

오이 덩굴쪼김병에 대한 효율적인 저항성 검정 방법

본 실험은 Fusarium oxysporum f. sp. cucumerinum에 의해 발생하는 오이 덩굴쪼김병의 효율적인 저항성 검정 방법을 확립하기 위하여 수행되었다. ITS와 TEF 유전자 염기서열 분석과 멜론, 참외, 오이 및 수박을 포함한 박과 작물에 대한 기주 특이성 분석을 통해 KR5 균주를 F. oxysporum f. sp. cucumerinum으로 동정하였다. 그리고 덩굴쪼김병균 KR5에 저항성 정도가 다른 오이 네 품종과 오이용 대목 두 품종을 선발하여 유묘의 생육 시기, 뿌리 상처, 접종원 농도 및 접종 후 재배 온도에 따라 이들 여섯 품종의 덩굴쪼김병 발생을 조사하였다. 저항성 품종의 덩굴쪼김병 발병도는 접종 후 재배 온도에 따라 차이를 나타냈으며, 저항성 품종들은 접종 후에 <TEX>$20^{\circ}C$</TEX>보다 <TEX>$25^{\circ}C$</TEX>나 <TEX>$30^{\circ}C$</TEX>에서 재배하였을 때 높은 저항성을 보였다. 그리고 유묘의 생육 시기 중 파종 후 7일된 유묘에 덩굴쪼김병균을 접종하였을 때 가장 큰 저항성과 감수성 차이를 나타내었다. 이상의 결과로부터 오이 덩굴쪼김병에 대한 저항성을 효과적으로 검정하는 방법으로 파종하고 7일 동안 재배한 유묘의 뿌리로부터 흙을 제거하고, 이들을 <TEX>$1.0{\times}10^6-1.0{\times}10^7$</TEX> conidia/ml 농도의 F. oxysporum f. sp. cucumerinum 포자현탁액에 침지하여 접종한 후에 건전한 원예용 상토에 이식하고 <TEX>$25^{\circ}C$</TEX> 생육상에서 3주간 재배하는 방법을 제안한다. The study was performed to establish an efficient screening method for resistant cucumber to Fusarium oxysporum f. sp. cucumerinum. The isolate KR5 was identified as F. oxysporum f. sp. cucumerinum based on molecular analyses of ITS and TEF genes and host-specificity test on cucurbits including melon, oriental melon, cucumber, and watermelon. Then four cucumber and two rootstock cultivars showing different resistance degrees to the Fusarium wilt pathogen KR5 were selected. And development of Fusarium wilt of the six cultivars according to several conditions, including incubation temperature after inoculation, inoculum concentration, root wounding, and growth stages of seedlings, was investigated. Disease severity of Fusarium wilt on the resistant cultivars was changed with incubation temperatures after inoculation. The resistant cultivars showed the higher resistance when inoculated plants were kept at 25 or <TEX>$30^{\circ}C$</TEX> than at <TEX>$20^{\circ}C$</TEX>. Among four different growth stages of the seedlings, seven-day-old seedling represented the most difference of resistance and susceptibility to Fusarium wilt. From above results, we suggest that an efficient screening method for resistant cucumber to F. oxysporum f. sp. cucumerinum is to dip the non-cut roots of seven-day-old seedlings in spore suspension of <TEX>$1.0{\times}10^6-1.0{\times}10^7$</TEX> conidia/ml and to transplant the seedling into a non-infected soil, and then to incubate the inoculated plants in a growth room at <TEX>$25^{\circ}C$</TEX> for 3 weeks to develop Fusarium wilt.