Arabidopsis is emerging as a model system to study the biochemistry, biological functions, and evolution of plant terpene secondary metabolism. It was previously shown that the Arabidopsis genome ...contains over 30 genes potentially encoding terpene synthases (TPSs). Here we report the characterization of a monoterpene synthase encoded by two identical, closely linked genes, At3g25820 and At3g25830. Transcripts of these genes were detected almost exclusively in roots. An At3g25820/At3g25830 cDNA was expressed in Escherichia coli, and the protein thus produced was shown to catalyze the formation of 10 volatile monoterpenes from geranyl diphosphate, with 1,8-cineole predominating. This protein was therefore designated AtTPS-Cin. The purified recombinant AtTPS-Cin displayed similar biochemical properties to other known monoterpene synthases, except for a relatively low Km value for geranyl diphosphate of 0.2 micromolar. At3g25820/At3g25830 promoter activity, measured with a β-glucuronidase (GUS) reporter gene, was primarily found in the epidermis, cortex, and stele of mature primary and lateral roots, but not in the root meristem or the elongation zone. Although the products of AtTPS-Cin were not detected by direct extraction of plant tissue, the recent report of 1,8-cineole as an Arabidopsis root volatile (Steeghs M, Bais HP, de Gouw J, Goldan P, Kuster W, Northway M, Fall R, Vivanco JM 2004 Plant Physiol 135: 47-58) suggests that the enzyme products may be released into the rhizosphere rather than accumulated. Among Arabidopsis TPSs, AtTPS-Cin is most similar to the TPS encoded by At3g25810, a closely linked gene previously shown to be exclusively expressed in flowers. At3g25810 TPS catalyzes the formation of a set of monoterpenes that is very similar to those produced by AtTPS-Cin, but its major products are myrcene and (E)-β-ocimene, and it does not form 1,8-cineole. These data demonstrate that divergence of organ expression pattern and product specificity are ongoing processes within the Arabidopsis TPS family.
A global vertically resolved aerosol data set covering more than 10 years of observations at more than 20 measurement sites distributed from 63∘ N to 52∘ S and 72∘ W to 124∘ E has been achieved ...within the Raman and polarization lidar network PollyNET. This network consists of portable, remote-controlled multiwavelength-polarization-Raman lidars (Polly) for automated and continuous 24/7 observations of clouds and aerosols. PollyNET is an independent, voluntary, and scientific network. All Polly lidars feature a standardized instrument design with different capabilities ranging from single wavelength to multiwavelength systems, and now apply unified calibration, quality control, and data analysis. The observations are processed in near-real time without manual intervention, and are presented online at polly.tropos.de. The paper gives an overview of the observations on four continents and two research vessels obtained with eight Polly systems. The specific aerosol types at these locations (mineral dust, smoke, dust-smoke and other dusty mixtures, urban haze, and volcanic ash) are identified by their Ångström exponent, lidar ratio, and depolarization ratio. The vertical aerosol distribution at the PollyNET locations is discussed on the basis of more than 55 000 automatically retrieved 30 min particle backscatter coefficient profiles at 532 nm as this operating wavelength is available for all Polly lidar systems. A seasonal analysis of measurements at selected sites revealed typical and extraordinary aerosol conditions as well as seasonal differences. These studies show the potential of PollyNET to support the establishment of a global aerosol climatology that covers the entire troposphere.
This study investigates the fatigue behavior of small specimens of Ti‐6Al‐4 V produced using the additive manufacturing process cold metal fusion. As the process is still comparatively new, no data ...were available beforehand. Strain‐controlled fatigue tests were carried out on small specimens under alternating loads. The results are presented in the form of a stress–strain diagram according to Ramberg–Osgood and a strain–life curve according to Coffin–Manson. Comparison with results reported for similar specimens fabricated via the laser engineered net shaping (LENS) additive method shows comparable fatigue behavior. Analysis of the fracture surfaces shows a very mixed fracture pattern with an often mountainous surface, especially for specimens tested under high strain. This made the detection of crack initiation very difficult for those specimens. The analysis of specimens tested under low strain reveals large surface defects to be the crack initiators. Smaller secondary cracks form at internal crack initiation facets.
A global vertically resolved aerosol data set covering more than 10 years of observations at more than 20 measurement sites distributed from 63 degrees N to 52 degrees S and 72 degrees W to 124 ...degrees E has been achieved within the Raman and polarization lidar network Polly(NET). This network consists of portable, remote-controlled multiwavelength-polarization-Raman lidars (Polly) for automated and continuous 24/7 observations of clouds and aerosols. Polly(NET) is an independent, voluntary, and scientific network. All Polly lidars feature a standardized instrument design with different capabilities ranging from single wavelength to multiwavelength systems, and now apply unified calibration, quality control, and data analysis. The observations are processed in near-real time without manual intervention, and are presented online at polly.tropos.de. The paper gives an overview of the observations on four continents and two research vessels obtained with eight Polly systems. The specific aerosol types at these locations (mineral dust, smoke, dust-smoke and other dusty mixtures, urban haze, and volcanic ash) are identified by their Angstrom exponent, lidar ratio, and depolarization ratio. The vertical aerosol distribution at the Polly(NET) locations is discussed on the basis of more than 55 000 automatically retrieved 30 min particle backscatter coefficient profiles at 532 nm as this operating wavelength is available for all Polly lidar systems. A seasonal analysis of measurements at selected sites revealed typical and extraordinary aerosol conditions as well as seasonal differences. These studies show the potential of Polly(NET) to support the establishment of a global aerosol climatology that covers the entire troposphere.
This study investigates the fatigue behavior of small specimens of Ti‐6Al‐4 V produced using the additive manufacturing process cold metal fusion. As the process is still comparatively new, no data ...were available beforehand. Strain‐controlled fatigue tests were carried out on small specimens under alternating loads. The results are presented in the form of a stress–strain diagram according to Ramberg–Osgood and a strain–life curve according to Coffin–Manson. Comparison with results reported for similar specimens fabricated via the laser engineered net shaping (LENS) additive method shows comparable fatigue behavior. Analysis of the fracture surfaces shows a very mixed fracture pattern with an often mountainous surface, especially for specimens tested under high strain. This made the detection of crack initiation very difficult for those specimens. The analysis of specimens tested under low strain reveals large surface defects to be the crack initiators. Smaller secondary cracks form at internal crack initiation facets.
Highlights
Fatigue performance of cold metal fusion (CMF) manufactured parts is comparable with laser engineered net shaping (LENS) manufactured parts.
Numerous pores are present.
Analysis of specimens tested under low strain reveals large surface defects to be the crack initiators.
Pine-feeding bark beetles (Coleoptera: Scolytidae) interact chemically with their host pines (Coniferales: Pinaceae) via the behavioral, physiological, and biochemical effects of one class of ...isoprenoids, the monoterpenes and their derivatives. Pine monoterpenes occur in the oleoresin and function as behaviorally active kairomones for pine bark beetles and their predators, presenting a classic example of tri-trophic chemical communication. The monoterpenes are also essential co-attractants for pine bark beetle aggregation pheromones. Ironically, pine monoterpenes are also toxic physiologically to bark beetles at high vapor concentrations and are considered an important component of the defense of pines. Research over the last 30 years has demonstrated that some bark beetle aggregation pheromones arise through oxygenation of monoterpenes, linking pheromone biosynthesis to the host pines. Over the last 10 years, however, several frequently occurring oxygenated monoterpene pheromone components (e.g., ipsenol, ipsdienol and frontalin) have also been shown to arise through highly regulated de novo pathways in the beetles (reviewed in Seybold and Tittiger, 2003). The most interesting nexus between these insects and their plant hosts involves the late-stage reactions in the monoterpenoid biosynthetic pathway, during which isomeric dimethylallyl diphosphate and isopentenyl diphosphate are ultimately elaborated to stereospecific monoterpenes in the trees and to hydroxylated monoterpenes or bicyclic acetals in the insects. There is signal stereospecificity in both production of and response to the monoterpenoid aggregation pheromones of bark beetles and in response to␣the monoterpenes of the pines. In the California fivespined ips, Ips paraconfusus, we have discovered a number of cytochome P450 genes that have expression patterns indicating that they may be involved in detoxifying monoterpene secondary metabolites and/or biosynthesizing pheromone components. Both processes result in the production of oxygenated monoterpenes, likely with varying degrees of stereospecificity. A behavioral analysis of the stereospecific response of I. paraconfusus to its pheromone is providing new insights into the development of an efficacious bait for the detection of this polyphagous insect in areas outside the western United States. In contrast, a Eurasian species that has arrived in California, the Mediterranean pine engraver, Orthotomicus (Ips) erosus, utilizes both a monoterpenoid (ipsdienol) and a hemiterpenoid (2-methyl-3-buten-2-ol) in its pheromone blend. The stereospecificity of the response of O. erosus to the monoterpenoid appears to be the key factor to the improved potency of the attractant bait for this invasive species.
Expression of heparanase (HPSE) in tumor cells is strongly associated with invasion, metastasis and angiogenesis. It also plays a key role during pregnancy, in processes of implantation as well as ...placentation. Vascular endothelial growth factor (VEGF) and heparin are known to alter HPSE expression, with heparin given prophylactically to women with a history of placenta-mediated complications in subsequent pregnancies.
We examined the growth-modulatory effects of different concentrations of heparin and VEGF on the choriocarcinoma cell-line JEG-3 and the expression of heparanase under VEGF and heparin by proliferation assays, PCR, and western blot.
Proliferation of JEG-3 cells was induced by heparin in a dose-dependent manner, whereas highly concentrated VEGF led to a decreased cell proliferation. Both agents did not influence the HPSE-expression.
The presumed pregnancy-protecting effects of heparin may partially be due to an increase of trophoblast proliferation and not via regulation of HPSE expression.
Precision oncology based on specific molecular alterations requires precise and reliable detection of therapeutic targets in order to initiate the optimal treatment. In many European ...countries—including Germany—assays employed for this purpose are highly diverse and not prescribed by authorities, making inter-laboratory comparison difficult. To ensure reproducible molecular diagnostic results across many laboratories and different assays, ring trials are essential and a well-established tool. Here, we describe the design and results of the ring trial for the detection of therapeutically relevant
PIK3CA
hotspot mutations in HR+/HER2-breast cancer tissue and liquid biopsy (LB). For
PIK3CA
mutation detection in tissue samples, 43 of the 54 participants (80%) provided results compliant with the reference values. Participants using NGS-based assays showed higher success rate (82%) than those employing Sanger sequencing (57%). LB testing was performed with two reference materials differing in the length of the mutated DNA fragments. Most participants used NGS-based or commercial real-time PCR assays (70%). The 167 bp fragments led to a successful
PIK3CA
mutation detection by only 31% of participants whereas longer fragments of 490 bp were detectable even by non-optimal assays (83%). In conclusion, the first ring trial for
PIK3CA
mutation detection in Germany showed that
PIK3CA
mutation analysis is broadly established for tissue samples and that NGS-based tests seem to be more suitable than Sanger sequencing.
PIK3CA
mutation detection in LB should be carried out with assays specifically designed for this purpose in order to avoid false-negative results.
Abstract
Introduction
Supporting and counselling couples with fertility issues prior to starting ART is a multidisciplinary diagnostic and therapeutic challenge. The first German/Austrian/Swiss ...interdisciplinary S2k guideline on “Diagnosis and Therapy Before Assisted Reproductive Treatments (ART)” was published in February 2019. This guideline was developed in the context of the guidelines program of the German Society of Gynecology and Obstetrics (DGGG) in cooperation with the Swiss Society of Gynecology and Obstetrics (SGGG) and the Austrian Society of Gynecology and Obstetrics (OEGGG).
Aims
One third of the causes of involuntary childlessness are still unclear, even if the woman or man have numerous possible risk factors. Because the topic is still very much taboo, couples may be socially isolated and often only present quite late to a fertility center. At present, there is no standard treatment concept, as currently no standard multidisciplinary procedures exist for the diagnostic workup and treatment of infertility. The aim of this guideline is to provide physicians with evidence-based recommendations for counselling, diagnostic workup and treatment.
Methods
This S2k guideline was developed on behalf of the Guidelines Commission of the DGGG by representative members from different professional medical organizations and societies using a structured consensus process.
Recommendations
The first part of this guideline focuses on the basic assessment of affected women, including standard anatomical and endocrinological diagnostic procedures and examinations into any potential infections. Other areas addressed in this guideline are the immunological workup with an evaluation of the patientʼs vaccination status, an evaluation of psychological factors, and the collection of data relating to other relevant factors affecting infertility. The second part will focus on explanations of diagnostic procedures compiled in collaboration with specialists from other medical specialties such as andrologists, human geneticists and oncologists.
Zusammenfassung
Einleitung
Die Begleitung von Paaren mit unerfülltem Kinderwunsch vor einer ART ist eine multidisziplinäre diagnostische und therapeutische Herausforderung. Im Februar 2019 erschien die erste deutschsprachige interdisziplinäre S2k-Leitlinie für die „Diagnostik und Therapie vor einer assistierten reproduktionsmedizinischen Behandlung (ART)“. Die Leitlinienerstellung erfolgte im Rahmen des Leitlinienprogrammes der Deutschen Gesellschaft für Gynäkologie und Geburtshilfe (DGGG) in Kooperation mit der Schweizer Gesellschaft für Gynäkologie und Geburtshilfe (SGGG) und der Österreichischen Gesellschaft für Gynäkologie und Geburtshilfe (OEGGG).
Ziel
Ein Drittel der Ursachen für Kinderlosigkeit bleibt unklar, auch wenn es eine Vielzahl an möglichen Risikofaktoren aufseiten der Frau und des Mannes gibt. Aufgrund der immer noch vorhandenen Tabuisierung des Themas sind die Paare teilweise sozial isoliert und wenden sich oftmals erst spät an ein Kinderwunschzentrum. Derzeit besteht kein einheitliches Behandlungskonzept, da keine fächerübergreifenden Handlungsanweisungen zur Diagnostik und Therapie der Infertilität vorliegen. Ziel der Leitlinie ist es, dem behandelnden Arzt/Ärztin im Rahmen der Beratung, diagnostischen Abklärung und Behandlung evidenzbasierte Empfehlungen anzubieten.
Methoden
Diese S2k-Leitlinie wurde durch einen strukturierten Konsens von repräsentativen Mitgliedern verschiedener Fachgesellschaften im Auftrag der Leitlinienkommission der DGGG entwickelt.
Empfehlungen
In diesem 1. Teil wird die grundlegende Abklärung der Frau thematisiert, mitsamt der grundlegenden anatomischen, endokrinologischen und infektiologischen Diagnostik. Darüber hinaus werden die immunologische Diagnostik, wie etwa die Abklärung des Impfstatus, die Evaluation psychologischer Faktoren sowie die Erhebung weiterer relevanter Einflussfaktoren auf die Infertilität thematisiert. Im 2. Teil erfolgt die Erläuterung der Abklärung in Zusammenarbeit mit anderen Fachdisziplinen wie Andrologen, Humangenetikern und Onkologen.
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