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Developing 'climate smart forestry' (CSF) indicators in mountain forest regions requires collection and evaluation of local data and their attributes. Genetic resources are listed among the core indicators for forest biological diversity. This study is a report on the evaluation of the standing genetic diversity within and across 12 pure beech stands (Fagus sylvatica L.) established within the CLIMO (CLImate Smart Forestry in Mountain Regions) project, using nuclear microsatellite markers. The sampling sites were set along the species' distribution range, including the Balkan region and extending towards the Iberian Peninsula. Cores or leaves from 20 to 23 old, mature trees per plot were sampled for DNA analysis. Genetic diversity indices were high across the range ([H.sub.E] = 0.74-0.81) with the highest in the Bosnian Mountains. Genetic divergence increased significantly with the geographical distance (Mantel test: r = 0.81, p < 0.001). Most of the stands exhibited an excess of heterozygotes, with the highest value at the Hungarian site ([H.sub.O]/[H.sub.E] = 1.177), where beech persists close to the eastern xeric limit of the species' distribution. STRUCTURE revealed within-region differentiation in the Balkan Peninsula, where the Bulgarian stand was the most outstanding. The genetic parameters of each stand could be assessed as a resource for CSF indicators interpreted especially at the local level. Key words: Fagus sylvatica, heterozygosity, F statistics, geographical groups, environmental variables. Le developpement d'indicateurs de foresterie intelligente face au climat (FIFC) dans les regions forestieres montagneuses exige la collecte et revaluation de donnees locales et de leurs attributs. Les ressources genetiques sont considerees comme des indicateurs de base pour la diversite biologique de la foret. Cette etude est un rapport sur revaluation de la diversite genetique actuelle dans et parmi 12 peuplements purs de hetre commun (Fagus sylvatica L.) etablis dans le cadre du projet CLIMO (CLIMO (<>) a l'aide de marqueurs microsatellites nucleaires. Les sites d'echantillonnage ont ete etablis le long de l'aire de repartition de l'espece, incluant la region des Balkans et se prolongeant vers la peninsule iberique. Des carottes ou des feuilles prelevees sur 20 a 23 vieux arbres matures par place echantillon ont ete echantillonnees pour l'analyse de l'ADN. Les indices de diversite genetique etaient eleves ([H.sub.E] = 0,74-0,81) a travers l'aire de repartition; les plus eleves ont ete observes dans les montagnes de Bosnie. La divergence genetique augmentait significativement avec la distance geographique (test de Mantel : r = 0,81, p < 0,001). La plupart des peuplements avaient un exces d'heterozygotes : le site hongrois, ou le hetre persiste pres de la limite xerique orientale de l'aire de repartition de 1'espece, avait la valeur la plus elevee ([H.sub.O]/[H.sub.E] = 1,177). STRUCTURE a revele une differenciation au sein de la region dans la peninsule des Balkans, ou un peuplement bulgare etait le plus remarquable. Les parametres genetiques de chaque peuplement pouvaient etre evalues en tant ressource pour les indicateurs de FIFC interpretes particulierement a l'echelle locale. [Traduit par la Redaction] Mots-cles: Fagus sylvatica, heterozygotie, statistique F, groupes geographiques, variables environnementales., 1. Introduction Forests play an important role in the mitigation of the effects of climate change. Therefore, climate smart forestry (CSF) may significantly invigorate the positive effects of forests and [...]

Objective: This study investigated mobile health enabled surveillance in ototoxicity. Method: This was a longitudinal study of 32 participants receiving chemotherapy. Baseline and exit audiograms that included conventional and extended high frequency audiometry were recorded within the patient's treatment venue using a validated mobile health audiometer. Results: Average hearing thresholds at baseline were within the normal range (81.2 per cent left; 93.8 per cent right), reducing at exit testing (71.9 per cent left; 78.1 per cent right). Half of participants presented with a threshold shift according to ototoxicity monitoring criteria. The frequencies affected the most were between 4000 and 16 000 Hz, with left ears significantly more affected than right ears. Noise levels exceeded the maximum permissible ambient noise levels in up to 43.8 per cent of low frequencies (250–1000 Hz). Conclusion: Mobile health supported audiometry proved to be an efficacious tool for ototoxicity monitoring at the treatment venue. Changes in hearing ability over time could be tracked, improving surveillance in patients with full treatment schedules. [ABSTRACT FROM AUTHOR]

Background: Populations of Olea europaea subsp. europaea var. sylvestris, the ancestor of cultivated olives, are scattered across the Mediterranean Basin. However, after millennia of possible hybridization with cultivated varieties, the genetic identity of many of these populations remain questionable. In the southern Levant, the plausible primary domestication center of olives, many of the naturally growing olive (NGOs) are considered feral, having developed from nearby olive groves. Here, we investigated the genetic identity of NGOs population in the Carmel region, hypothesizing that their specific location, which limit anemophily, provided an opportunity for the persistence of genuine var. sylvestris., Results: We mapped more than 1,000 NGOs on the Kurkar ridge along the Carmel coast, within and outside the residential area of Atlit and used simple sequence repeats of 14 loci to assess the spatial genetic structure of 129 NGOs. Genetic diversity parameters and genetic distances between NGO and cultivated olives, as well as phenotypic and morphometric analyses of their oil content and pits, respectively, indicated the presence of a genuine var. sylvestris population. However, NGOs within the residential area of Atlit and old settlements showed an intermediate admix genetic structure, indicating on hybridization with local varieties, a consequence of their proximity to cultivated trees., Conclusions: Integrating the results of genetic and phenotypic analyses we provide crucial evidence of the presence of a genuine var. sylvestris population in the southern Levant, in close geographical proximity to archaeological sites with the earliest evidence of olive exploitation in the ancient world. We supplement the results with recommendations for a conservation program that combines municipal requirements and the urgent need to preserve the largest population of var. sylvestris in the southern Levant., (© 2024. The Author(s).)

Purpose: The objective of this study was to determine the normative vestibulo-ocular reflex gain output values of the computerized rotational head impulse test (crHIT) with stationary visual targets (earth bound) in healthy participants in each decade age band of life: 10-19, 20-29, 30-39, 40-49, 50-59, 60-69, and 70+ years., Method: Seventy-seven community-dwelling participants (10-85 years of age) with normal lateral semicircular canal (SCC) functioning and no symptoms or history of vestibular dysfunction were recruited through convenience sampling and assessed with the crHIT using stationary targets. These participants were assessed using two standard protocols in a randomized order., Results: Results from 77 participants ( M age = 46 years; 43 women, 34 men) were analyzed. Pearson's correlation coefficient and simple linear regression indicated a statistically significant relationship between crHIT gain output and age ( p > .05) for right gain, 1030°/s 2 , and left gain, 1005°/s 2 . Although a statistically significant relationship was found, the slope was minor, demonstrating that the clinical effect of age on crHIT gain output was insignificant. Furthermore, no statistically significant relationship exists between crHIT gain output and gender ( p > .05). Age-dependent normative data were calculated using the 2.5th and 97.5th confidence interval (CI) percentile method. The responses of angular vestibulo-ocular reflex (aVOR) gain values for crHIT are expected to occur within the range for lower limit reference interval (RI) of 0.85-0.9 and upper limit RI of 1.11-1.18 for 1030°/s 2 and lower limit RI of 0.86-0.92 and upper limit RI of 1.13-1.16 for 1005°/s 2 . It can be expected that 90% CI of the population with normal lateral SCC functioning will have aVOR gain values that fall within this range., Conclusion: Despite a statistically significant relationship that exists with aVOR gain output and age, the changes are minor, declining by 0.0088 units per 10 years, justifying the same normative data for all decade age bands.

Abstract: We report on an adrenocortical carcinoma (ACC) patient, which has exhausted previous treatment options and was scheduled for prostate-specific membrane antigen (PSMA)- and C-X-C motif chemokine receptor 4 (CXCR4)-targeted PET/CT. We identified PSMA-avid pulmonary metastases exhibiting modest radiotracer accumulation, while chemokine receptor PET/CT provided intense uptake. This dual-tracer molecular imaging approach revealed that chemokine receptor PET appears to be more suitable in patients with advanced ACC, indicating that CXCR4-directed radioligand therapy may be considered in such patients suffering from end-stage disease. Given its dismal prognosis, chemokine receptor-directed theranostics may therefore extend the therapeutic armamentarium as last-line option in advanced ACC., Competing Interests: Conflicts of interest and sources of funding: This study was partially supported by the Okayama University “RECTOR” Program, KAKENHI grant (22H03027) from the Japan Society for the Promotion of Science (T.H.), and the German Research Foundation (453989101, R.A.W., T.H.; 507803309, R.A.W.). This work was supported by the Deutsche Forschungsgemeinschaft (DFG AL 203/1-1 and CRC/Transregio 205 “The Adrenal: Central Relay in Health and Disease”). R.A.W. and A.K.B. have received speaker honoraria from Novartis/AAA and PentixaPharm. R.A.W. reports advisory board work for Novartis/AAA and Bayer. A.K.B. is a member of the advisory board of PentixaPharm. All other authors declare no conflict of interest., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Background: Despite the increasing number of epigenomic studies in plants, little is known about the forces that shape the methylome in long-lived woody perennials. The Lombardy poplar offers an ideal opportunity to investigate the impact of the individual environmental history of trees on the methylome., Results: We present the results of three interconnected experiments on Lombardy poplar. In the first experiment, we investigated methylome variability during a growing season and across vegetatively reproduced generations. We found that ramets collected over Europe and raised in common conditions have stable methylomes in symmetrical CG-contexts. In contrast, seasonal dynamics occurred in methylation patterns in CHH context. In the second experiment, we investigated whether methylome patterns of plants grown in a non-parental environment correlate with the parental climate. We did not observe a biological relevant pattern that significantly correlates with the parental climate. Finally, we investigated whether the parental environment has persistent carry-over effects on the vegetative offspring's phenotype. We combined new bud set observations of three consecutive growing seasons with former published bud set data. Using a linear mixed effects analysis, we found a statistically significant but weak short-term, parental carry-over effect on the timing of bud set. However, this effect was negligible compared to the direct effects of the offspring environment., Conclusions: Genome-wide cytosine methylation patterns in symmetrical CG-context are stable in Lombardy poplar and appear to be mainly the result of random processes. In this widespread poplar clone, methylation patterns in CG-context can be used as biomarkers to infer a common ancestor and thus to investigate the recent environmental history of a specific Lombardy poplar. The Lombardy poplar shows high phenotypic plasticity in a novel environment which enabled this clonal tree to adapt and survive all over the temperate regions of the world., (© 2024. The Author(s).)

The germline TP53 p.R337H mutation is reported as the most common germline TP53 variant. It exists at a remarkably high frequency in the population of southeast Brazil as founder mutation in two distinct haplotypes with the most frequent co-segregating with the p.E134∗ variant of the XAF1 tumor suppressor and an increased cancer risk. Founder mutations demonstrate linkage disequilibrium with neighboring genetic polymorphic markers that can be used to identify the founder variant in different geographic regions and diverse populations. We report here a shared haplotype among Brazilian, Portuguese, and Spanish families and the existence of three additional distinct TP53 p.R337H alleles. Mitochondrial DNA sequencing and Y-STR profiling of Brazilian carriers of the founder TP53 p.R337H allele reveal an excess of Native American haplogroups in maternal lineages and exclusively European haplogroups in paternal lineages, consistent with communities established through male European settlers with extensive intermarriage with Indigenous women. The identification of founder and independent TP53 p.R337H alleles underlines the importance for considering the haplotype as a functional unit and the additive effects of constitutive polymorphisms and associated variants in modifier genes that can influence the cancer phenotype., Competing Interests: Declaration of interests K.B. reports an immediate family member on the scientific advisory board of Emendo Biotherapeutics, Karyopharm Therapeutics, Imago BioSciences, and DarwinHealth; is co-founder of Isabl Technologies; and has equity interest in Imago BioSciences, Emendo Biotherapeutics, and Isabl Technologies., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Objectives: To determine the test-retest reliability of cervical and ocular vestibular-evoked myogenic potentials (c&oVEMP) evoked by 500 Hz narrowband (NB) CE-Chirp and broadband (BB) CE-Chirp stimuli., Design: Twenty healthy participants (10 female) were tested twice on the same day to determine the within-session reliability and 1 week later to determine the between-session reliability. The latency, amplitude, and asymmetry ratio of c&oVEMPs elicited by 95 dB nHL air-conducted (AC) 500 Hz NB CE-Chirp and BB CE-Chirp were recorded bilaterally., Results: A moderate to good between-session reliability with intraclass correlation coefficient (ICC) values ranging from 0.52 to 0.82 was observed for cVEMP latency, amplitude, and asymmetry ratio evoked by 500 Hz NB CE-Chirp, as well as for the BB CE-Chirp cVEMP amplitude (ICC of 0.70 and 0.84). In contrast, an overall poor reliability ICC values between 0.30 and 0.42 for cVEMP latency and asymmetry ratio were observed for BB CE-Chirp. For the oVEMP, overall poor between-session reliability for all response parameters evoked by the 500 Hz NB CE-Chirp and the BB CE-Chirp was observed., Conclusions: The 500 Hz NB CE-Chirp was more reliable than the BB CE-Chirp in terms of cVEMP latency, amplitude, and asymmetry ratio. Further investigation using the standard electrode montage is necessary to assess the test-retest reliability of the chirp-evoked oVEMP., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

The enzyme aldosterone synthase (CYP11B2) is specifically expressed in aldosterone-producing tissue of the adrenal cortex and is overexpressed in aldosterone-producing adenomas (APA). It therefore represents an ideal target for molecular imaging, particularly for the differential diagnosis between bilateral hyperplasia and unilateral APA in primary aldosteronism. However, the presence of the cortisol-producing enzyme 11β-hydroxylase (CYP11B1) in the adrenal cortex remains very challenging owing to its high homology to CYP11B2. Within this study, we efficiently synthesized a variety of disubstituted fluorinated pyridines and pyrazines by Suzuki coupling reactions. These compounds were evaluated for their ability to inhibit CYP11B1 and CYP11B2 in transfected Y1 cells and in NCI-h295 cells. Several compounds were found to exhibit excellent affinity (IC 50  < 10 nM) to CYP11B2 as well as strong selectivity (up to 125-fold) over CYP11B1. These findings support the further development of an analogous 18 F-labelled PET tracer., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: AS, BH and SH filed a patent application: Compound for in vivo diagnosis of a dysfunction of adrenal glands. WO/2018/141541. All other authors declare no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objectives: To compare the response rate and response parameters of cervical and ocular vestibular evoked myogenic potentials (c&oVEMP) elicited by narrowband (NB) and broadband (BB) CE-Chirp, with the more classical tone burst (TB) and click VEMPs., Design: The response rate, latency, amplitude and asymmetry ratio of c&oVEMPs elicited by 95 dB nHL air conducted (AC) 500 Hz NB CE-chirp, BB CE-chirp, 500 Hz TB and click stimuli were recorded bilaterally., Study Sample: 20 male and 38 female participants (19-39 years)., Results: For the cVEMP, the highest response rate was found for NB chirp (100%), followed by TB (91%), BB chirp (87%) and finally click (85%). A similar order was seen for oVEMP with percentages of 100%; 57%, 57%, and 43%. The 500 Hz NB CE-Chirp elicited significantly shorter cVEMP P1 and N1 latencies and significantly larger c&oVEMP amplitudes compared to all other stimuli. BB CE-Chirp elicited significantly shorter c&oVEMP P1 and N1 latencies with smaller amplitudes compared to TB. Asymmetry ratios were not statistically significant for all comparisons., Conclusion: The 500 Hz NB CE-chirp provides the highest response rates, shorter latencies and larger amplitudes, and therefore seem a promising stimulus for reliably measuring c&oVEMPs in clinical practice.

Omnivorous and opportunistic species may be good indicators of food availability. Gulls often use human-impacted landscapes and may respond to changes by altering their feeding ecology. We investigated the foraging behavior of individual common gulls ( Larus canus ), focusing on their distribution during foraging and their selected habitat types. We tracked adult common gulls using GPS telemetry at their largest breeding colony in the southwestern Baltic Sea, Germany. Foraging habitats were analyzed from tracking data for three breeding seasons 2016, 2017, and 2019 and were compared with potentially available foraging habitats. Most breeding birds flew toward terrestrial areas. Feeding sites were located on average 11.7-14.3 km from the colony (range 0.9-36.5 km). Corn and sugar beet fields were used significantly and extensively compared with their availability in 2016 and 2017, while wheat, rape, and barley fields were used significantly less. Data from 2019 suggested seasonal shifts in habitat use. Birds spent between 30 and 1300 min per week at their preferred feeding sites, with significant differences between the major habitats selected. We found a stable, clear, multiyear pattern in common gull foraging behavior in relation to agricultural practices. Fields with little or no crop cover and thus access to the soil were preferred over fields with high crop cover. These results suggest that local food availability may be limiting further population increases in this species., Competing Interests: The authors declare that they have no conflict of interest with the content of this article., (© 2022 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Objectives: This study investigated the changes in vestibular and cochlear function in patients receiving platinum-based chemotherapy., Methods: A longitudinal study of 32 participants (10-70 years) receiving chemotherapy participated in the study. Baseline and exit vestibular and hearing assessments that included video head impulse (VHIT) testing, cervical and ocular vestibular evoked myogenic potentials (VEMP), dynamic visual acuity (DVA) and pure-tone audiometry were performed at the patient's treatment venue., Results: Half (50%) of the participants showed cochleotoxicity from baseline to exit testing, with left ears significantly more affected than right ears. There was no consistent relationship between hearing loss and vestibular dysfunction. DVA yielded normal results at baseline and exit testing in all participants. VEMP responses were absent in 28.1% of participants at baseline, reflecting the challenges of using VEMP for monitoring. VEMP and VHIT results showed a statistically significant ( p < 0.05) decline in results from baseline to exit testing; however, participants did not report symptoms related to vestibular dysfunction. VHIT also showed left ears significantly ( p < 0.05) more affected than right ears., Conclusion: VHIT proved to be a valuable measure of changes in vestibular function secondary to ototoxicity. Future investigations should determine vestibulotoxicity criteria and optimal protocols for vestibulotoxicity monitoring at the patient's treatment venue., Competing Interests: Declaration of conflicting interestsThe author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: DS has equity interest, consultancy and potential royalties in the hearX Group.

Context: Adrenocortical carcinoma (ACC) is a rare endocrine malignancy with limited treatment options. Theranostic approaches with adrenal specific radiotracers hold promise for improved diagnostics and treatment., Objective: Here, we report a new theranostic approach to advanced ACC applying (R)-1-[1-(4-[123I]iodophenyl)ethyl]-1H-imidazole-5-carboxylic acid azetidinyl amide ([123I]IMAZA) for diagnostic imaging and [131I]IMAZA for radionuclide therapy., Methods: Sixty-nine patients with nonresectable, metastatic ACCs were screened using a diagnostic [123I]IMAZA scan. Patients with significant uptake in all tumoral lesions were offered treatment with [131I]IMAZA. Tumor response was assessed according to Response Evaluation Criteria in Solid Tumors (RECIST version 1.1), and adverse effects were assessed by Common Toxicity Criteria (version 5.0)., Results: After screening, 13 patients were treated with a median of 25.7 GBq [131I]IMAZA (range 18.1-30.7 GBq). Five individuals received a second treatment course. Best response was a decrease in the RECIST target lesions of -26% in 2 patients. Five patients with disease stabilization experienced a median progression-free survival of 14.3 months (range 8.3-21.9). Median overall survival in all patients was 14.1 months (4.0-56.5) after therapy. Treatment was well tolerated, in other words no severe toxicities (CTCAE grade ≥3) were observed., Conclusion: In patients with advanced ACC refractory to standard therapeutic regimens, [131I]IMAZA treatment was associated with disease stabilization and nonsignificant tumor size reduction in a significant patient fraction and only limited toxicities. High [131I]IMAZA-uptake in tumor lesions was observed in 38.5% of patients with advanced ACC, rendering [131I] IMAZA a potential treatment option in a limited, well-defined patient fraction. Further clinical trials will be necessary to evaluate the full potential of this novel theranostic approach., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Endocrine Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: National information regarding ototoxicity monitoring practices are limited for patients undergoing chemotherapy in South Africa., Objectives: To determine (1) the national status of ototoxicity monitoring implemented in private and public cancer facilities, (2) the knowledge and ototoxicity monitoring approaches implemented, and (3) reported challenges., Method: A descriptive quantitative survey was conducted in public and private oncology units and audiology referral clinics. Private (60%) and public (43%) oncology units that provide platinum-based chemotherapy in South Africa and audiology referral units (54%) were: (1) surveyed telephonically to determine if ototoxicity monitoring takes place; and (2) a self-administered survey was sent to qualifying oncology units and audiology referral clinics., Results: All public oncology units reported that ototoxicity monitoring only occurs on referral and is not standard practice. All private oncology units indicated that monitoring is on a patient self-referral basis when symptoms occur. Poor awareness of ototoxicity monitoring best practice guidelines was reported by all oncology units and 14% of audiology referral clinics. Audiology referral clinics reported adequate knowledge of ototoxicity protocols although they are not widely used with only 43% following best practice guidelines. The most prominent challenges reported by participants was referral system (67% oncology units; 57% audiology referral clinics), environmental noise (83% oncology units; 86% audiology referral clinics) and the compromised status of cancer patients (67% oncology units; 57% audiology referral clinics)., Conclusion: Ototoxicity monitoring is not routinely implemented across oncology units in South Africa. Multidisciplinary teamwork and a simplified national ototoxicity monitoring protocol may improve hearing outcomes for patients.

Purpose: Adrenal tumors represent a diagnostic and therapeutic challenge. Promising results have been obtained through targeting the cytochrome P450 enzymes CYP11B1 and CYP11B2 for molecular imaging, and [ 123/131 I]iodometomidate ([ 123/131 I]IMTO) has even been successfully introduced as a theranostic agent. As this radiopharmaceutical shows rapid metabolic inactivation, we aimed at developing new improved tracers., Methods: Several IMTO derivatives were newly designed by replacing the unstable methyl ester by different carboxylic esters or amides. The inhibition of aldosterone and cortisol synthesis was tested in different adrenocortical cell lines. The corresponding radiolabeled compounds were assessed regarding their stability, in vitro cell uptake, in vivo biodistribution in mice, and their binding specificity to cryosections of human adrenocortical and non-adrenocortical tissue. Furthermore, a first investigation was performed in patients with known metastatic adrenal cancer using both [ 123 I]IMTO and the most promising compound (R)-1-[1-(4-[ 123 I]iodophenyl)ethyl]-1H-imidazole-5-carboxylic acid azetidinylamide ([ 123 I]IMAZA) for scintigraphy. Subsequently, a first endoradiotherapy with [ 131 I]IMAZA in one of these patients was performed., Results: We identified three analogues to IMTO with high-affinity binding to the target enzymes and comparable or higher metabolic stability and very high and specific accumulation in adrenocortical cells in vitro and in vivo. Labeled IMAZA exhibited superior pharmacokinetic and imaging properties compared to IMTO in mice and 3 patients, too. An endoradiotherapy with [ 131 I]IMAZA induced a 21-month progression-free interval in a patient with rapidly progressing ACC prior this therapy., Conclusion: We developed the new radiopharmaceutical [ 123/131 I]IMAZA with superior properties compared to the reference compound IMTO and promising first experiences in humans., (© 2021. The Author(s).)

Background: The chemokine receptor CCR7 is crucial for an intact immune function, but its expression is also associated with clinical outcome in several malignancies. No data exist on the expression of CCR7 in adrenocortical tumors., Methods: CCR7 expression was investigated by qRT-PCR and immunohistochemistry in 4 normal adrenal glands, 59 adrenocortical adenomas, and 181 adrenocortical carcinoma (ACC) samples., Results: CCR7 is highly expressed in the outer adrenocortical zones and medulla. Aldosterone-producing adenomas showed lower CCR7 protein levels (H-score 1.3 ± 1.0) compared to non-functioning (2.4 ± 0.5) and cortisol-producing adenomas (2.3 ± 0.6), whereas protein expression was variable in ACC (1.8 ± 0.8). In ACC, CCR7 protein expression was significantly higher in lymph node metastases (2.5 ± 0.5) compared to primary tumors (1.8±0.8) or distant metastases (2.0 ± 0.4; p < 0.01). mRNA levels of CCR7 were not significantly different between ACCs, normal adrenals, and adrenocortical adenomas. In contrast to other tumor entities, neither CCR7 protein nor mRNA expression significantly impacted patients' survival., Conclusion: We show that CCR7 is expressed on mRNA and protein level across normal adrenals, benign adrenocortical tumors, as well as ACCs. Given that CCR7 did not influence survival in ACC, it is probably not involved in tumor progression, but it could play a role in adrenocortical homeostasis.

In this chapter, frequently used methods for elucidating sequence and structure of chloroplast genomes are reviewed, as a current best practice guide. This concerns methods for DNA extraction, sequencing library preparation, and bioinformatics (assembly, verification, annotation, and sequence comparisons). Recommendations for standard data reporting practices are given-chloroplast genome sequencing reports can be highly formalized, and publication in the form of standard data reports is the best option for comparison and meta-analysis purposes.

Chemokine receptors have a negative impact on tumor progression in several human cancers and have therefore been of interest for molecular imaging and targeted therapy. However, their clinical and prognostic significance in adrenocortical carcinoma (ACC) is unknown. The aim of this study was to evaluate the chemokine receptor profile in ACC and to analyse its association with clinicopathological characteristics and clinical outcome. A chemokine receptor profile was initially evaluated by quantitative PCR in 4 normal adrenals, 18 ACC samples and human ACC cell line NCI-H295. High expression of CXCR4 and CXCR7 in both healthy and malignant adrenal tissue and ACC cells was confirmed. In the next step, we analyzed the expression and cellular localization of CXCR4 and CXCR7 in ACC by immunohistochemistry in 187 and 84 samples, respectively. These results were correlated with clinicopathological parameters and survival outcome. We detected strong membrane expression of CXCR4 and CXCR7 in 50% of ACC samples. Strong cytoplasmic CXCR4 staining was more frequent among samples derived from metastases compared to primaries ( p= 0.01) and local recurrences ( p= 0.04). CXCR4 membrane staining positively correlated with proliferation index Ki67 (r=0.17, p= 0.028). CXCR7 membrane staining negatively correlated with Ki67 (r=-0.254, p= 0.03) but positively with tumor size (r=0.3, p= 0.02). No differences in progression-free or overall survival were observed between patients with strong and weak staining intensities for CXCR4 or CXCR7. Taken together, high expression of CXCR4 and CXCR7 in both local tumors and metastases suggests that some ACC patients might benefit from CXCR4/CXCR7-targeted therapy., (Copyright © 2020 Chifu, Heinze, Fuss, Lang, Kroiss, Kircher, Ronchi, Altieri, Schirbel, Fassnacht and Hahner.)

Over the last several decades, several lines of evidence have shown that epigenetic modifications modulate phenotype and mediate an organism's response to environmental stimuli. Plant DNA is normally highly methylated, although notable differences exist between species. Many biomolecular techniques based on PCR have been developed to analyse DNA methylation status, however a qualitative leap was made with the advent of next-generation sequencing (NGS). In the case of large, repetitive, or not-yet-sequenced genomes characterised by a high level of DNA methylation, the NGS analysis of bisulphite pre-treated DNA is expensive and time consuming, and moreover, in some cases data analysis is a major challenge. Methylation-sensitive amplification polymorphism (MSAP) analysis is a highly effective method to study DNA methylation. The method is based on the comparison of double DNA digestion profiles ( Eco RI- Hpa II and Eco RI- Msp I) to reveal methylation pattern variations. These are often attributable to pedoclimatic and stress conditions which affect all organisms during their lifetime. In our study, five white poplar ( Populus alba L.) specimens were collected from different monoclonal stands in the Maltese archipelago, and their DNA was processed by means of an innovative approach where MSAP analysis was followed by NGS. This allowed us to identify genes that were differentially methylated among the different specimens and link them to specific biochemical pathways. Many differentially methylated genes were found to encode transfer RNAs (tRNAs) related to photosynthesis or light reaction pathways. Our results clearly demonstrate that this combinatorial method is suitable for epigenetic studies of unsequenced genomes like P. alba (at the time of study), and to identify epigenetic variations related to stress, probably caused by different and changing pedoclimatic conditions, to which the poplar stands have been exposed.

Massively parallel sequencing of cDNA is an efficient route for generating sequence collections that represent expressed genes under different environmental control. The analysis of their sequence helps in developing molecular markers, such as SNPs, which represent a useful tool in detecting adaptive signals in populations. In this study novel PCR markers, based on stress responsive genes, were designed from the transcriptome of the haploxylon Swiss stone pine (Pinus cembra L.) and tested for SNPs in the diploxylon Scots pine (Pinus sylvestris L.). 84 primers were tested on P. sylvestris DNA samples originating from three different types of habitat. After sequencing and BLAST search of the amplified products, parts of 19 different candidate genes were analysed by considering the polymorphic sites, insertions/deletions as well as synonymous and non-synonymous SNPs. In a total of 3735 sites no indels, eight synonymous and 11 non-synonymous SNPs were found. By providing de novo molecular markers developed in P. cembra and tested for transferability in Scots pine, our results give support for the use of de novo markers targeting conserved regions across different pines. The SNPs detected may have important applications in further studies of adaptive genetic variation, providing tools to study relevant genes important in the long-term adaptation of pine species.

Genome mining identified the fungal-bacterial endosymbiosis Rhizopus microsporus - Mycetohabitans (previously Burkholderia ) rhizoxinica as a rich source of novel natural products. However, most of the predicted compounds have remained cryptic. In this study, we employed heterologous expression to isolate and characterize three ribosomally synthesized and post-translationally modified peptides with lariat topology (lasso peptides) from the endosymbiont M . rhizoxinica : burhizin-23, mycetohabin-16, and mycetohabin-15. Through coexpression experiments, it was shown that an orphan gene product results in mature mycetohabin-15, albeit encoded remotely from the core biosynthetic gene cluster. Comparative genomics revealed that mycetohabins are highly conserved among M . rhizoxinica and related endosymbiotic bacteria. Gene knockout and reinfection experiments indicated that the lasso peptides are not crucial for establishing symbiosis; instead, the peptides are exported into the environment during endosymbiosis. This is the first report on lasso peptides from endosymbiotic bacteria.

Silver fir ( Abies alba Mill.) is a keystone conifer of European montane forest ecosystems that has experienced large fluctuations in population size during during the Quaternary and, more recently, due to land-use change. To forecast the species' future distribution and survival, it is important to investigate the genetic basis of adaptation to environmental change, notably to extreme events. For this purpose, we here provide a first draft genome assembly and annotation of the silver fir genome, established through a community-based initiative. DNA obtained from haploid megagametophyte and diploid needle tissue was used to construct and sequence Illumina paired-end and mate-pair libraries, respectively, to high depth. The assembled A. alba genome sequence accounted for over 37 million scaffolds corresponding to 18.16 Gb, with a scaffold N50 of 14,051 bp. Despite the fragmented nature of the assembly, a total of 50,757 full-length genes were functionally annotated in the nuclear genome. The chloroplast genome was also assembled into a single scaffold (120,908 bp) that shows a high collinearity with both the A. koreana and A. sibirica complete chloroplast genomes. This first genome assembly of silver fir is an important genomic resource that is now publicly available in support of a new generation of research. By genome-enabling this important conifer, this resource will open the gate for new research and more precise genetic monitoring of European silver fir forests., (Copyright © 2019 Mosca et al.)

Background: Decentralized detection and monitoring of hearing loss can be supported by new mobile health technologies using automated testing that can be facilitated by minimally trained persons. These may prove particularly useful in an infectious disease (ID) clinic setting where the risk of hearing loss is high., Purpose: To evaluate the clinical utility of mobile and automated audiometry hearing health technology in an ID clinic setting., Research Design: Smartphone-automated pure-tone audiometry (PTA) (hearTest™) and speech-in-noise testing (SA English digits-in-noise [DIN] test) were compared with manual audiometry (2, 4, and 8 kHz). Smartphone-automated PTA and the DIN test were repeated to determine the test-retest reliability., Study Sample: Two hundred subjects (73% female and 27% male) were enrolled. Fifty participants were retested with the smartphone applications. Participants ranged from an age of 18 to 55 years with a mean age of 44.4 (8.7 standard deviation)., Data Analysis: Threshold comparisons were made between smartphone audiometry testing and manual audiometry. Smartphone-automated PTA, manual audiometry, and test-retest measures were compared (Wilcoxon signed ranked test). Spearman rank correlation test was used to determine the relationship between the smartphone applications and manual audiometry, as well as for test-retest reliability., Results: Within all participants, 88.2% of thresholds corresponded within 10 dB or less between smartphone audiometry and manual audiometry. There was a significant difference (p < 0.05) between the right ear at 4 and 8 kHz and in the left ear at 2 and 4 kHz between smartphone and manual audiometry, respectively. No significant difference was noted (p < 0.05) between test and retest measures of smartphone technology., Conclusions: Smartphone audiometry with calibrated headphones provides reliable results in an ID clinic setting and can be used as a baseline and monitoring tool at ID clinics., (American Academy of Audiology.)

More frequently occurring, drought waves call for a deeper understanding of tree hydraulics and fast and easily applicable methods to measure drought stress. The aim of this study was to establish empirical relationships between the percent loss of hydraulic conductivity (PLC) and the relative water loss (RWL) in woody stem axes with different P 50 , i.e. the water potential (Ψ) that causes 50% conductivity loss. Branches and saplings of temperate conifer (Picea abies, Larix decidua) and angiosperm species (Acer campestre, Fagus sylvatica, Populus x canescens, Populus tremula, Sorbus torminalis) and trunk wood of mature P. abies trees were analyzed. P 50 was calculated from hydraulic measurements following bench top dehydration or air injection. RWL and PLC were fitted by linear, quadratic or cubic equations. Species- or age-specific RWLs at P 50 varied between 10 and 25% and P 88 , the Ψ that causes 88% conductivity loss, between 18 and 44%. P 50 was predicted from the relationship between Ψ and the RWL. The predictive quality for P 50 across species was almost 1:1 (r 2  = 0.99). The approach presented allows thus reliable and fast prediction of PLC from RWL. Branches and saplings with high hydraulic vulnerability tended to have lower RWLs at P 50 and at P 88 . The results are discussed with regard to the different water storage capacities in sapwood and survival strategies under drought stress. Potential applications are screening trees for drought sensitivity and a fast interpretation of diurnal, seasonal or drought induced changes in xylem water content upon their impact on conductivity loss., (© 2018 The Authors. Physiologia Plantarum published by John Wiley & Sons Ltd on behalf of Scandinavian Plant Physiology Society.)

In the absence of genetic diversity, plants rely on the capacity of phenotypic plasticity to cope with shifts in environmental conditions. Understanding the mechanisms behind phenotypic plasticity and how local phenotypic adjustments are transferred to clonal offspring, will provide insight into its ecological and evolutionary significance. Epigenetic changes have recently been proposed to play a crucial role in rapid environmental adaptation. While the contribution of epigenetic changes to phenotypic plasticity has been extensively studied in sexual reproducing model organisms, little work has been done on vegetative generations of asexual reproducing plant species. We studied the variability of DNA methylation and bud set phenology of the Lombardy poplar ( Populus nigra cv. Italica Duroi), a cultivated tree representing a single genotype worldwide distributed since the eighteenth century. Bud set observations and CpG methyl polymorphisms were studied on vegetative offspring resulting from cuttings grown for one season in a common glasshouse environment. The cuttings were collected from 60 adult Lombardy poplars growing in different environments. The physiological condition of the cuttings was determined by measuring weight and nutrient condition. Methylation sensitive amplified polymorphisms were used to obtain global patterns of DNA methylation. Using logistic regression models, we investigated correlations among epigenotype, bud phenology, and the climate at the home site of the donor trees, while accounting for physiological effects. We found significant epigenetic variation as well as significant variation in bud phenology, in the absence of genetic variation. Remarkably, phenology of bud set observed at the end of the growing season in the common environment was significantly correlated with climate variables at the home site of the mother trees, specifically the average temperature of January and monthly potential evapotranspiration. Although we could not directly detect significant effects of epigenetic variation on phenology, our results suggest that, in the Lombardy poplar, epigenetic marks contribute to the variation of phenotypic response that can be transferred onto asexually reproduced offspring resulting in locally adapted ecotypes. This contributes to the growing evidence that epigenetic-based transgenerational inheritance might be relevant for adaptation and evolution in contrasting or rapidly changing environments.

Primary aldosteronism is the most frequent cause of secondary hypertension and is associated with increased morbidity and mortality compared with hypertensive controls. The central diagnostic challenge is the differentiation between bilateral and unilateral disease, which determines treatment options. Bilateral adrenal venous sampling, currently recommended for differential diagnosis, is an invasive procedure with several drawbacks, making it desirable to develop novel noninvasive diagnostic tools. When investigating the expression pattern of chemokine receptors by quantitative real-time polymerase chain reaction and immunohistochemistry, we observed high expression of CXCR4 (CXC chemokine receptor type 4) in aldosterone-producing tissue in normal adrenals, adjacent adrenal cortex from adrenocortical adenomas, and in aldosterone-producing adenomas (APA), correlating strongly with the expression of CYP11B2 (aldosterone synthase). In contrast, CXCR4 was not detected in the majority of nonfunctioning adenomas that are frequently found coincidently. The specific CXCR4 ligand 68Ga-pentixafor has recently been established as radiotracer for molecular imaging of CXCR4 expression and showed strong and specific binding to cryosections of APAs in our study. We further investigated 9 patients with primary aldosteronism because of APA by 68Ga-pentixafor-positron emission tomography. The tracer uptake was significantly higher on the side of increased adrenocortical aldosterone secretion in patients with APAs compared with patients investigated by 68Ga-pentixafor-positron emission tomography for other causes. Molecular imaging of aldosterone-producing tissue by a CXCR4-specific ligand may, therefore, be a highly promising tool for noninvasive characterization of patients with APAs., (© 2017 American Heart Association, Inc.)

Background: Picea chihuahuana , which is endemic to Mexico, is currently listed as "Endangered" on the Red List. Chihuahua spruce is only found in the Sierra Madre Occidental (SMO), Mexico. About 42,600 individuals are distributed in forty populations. These populations are fragmented and can be classified into three geographically distinct clusters in the SMO. The total area covered by P. chihuahuana populations is less than 300 ha. A recent study suggested assisted migration as an alternative to the ex situ conservation of P. chihuahuana , taking into consideration the genetic structure and diversity of the populations and the predictions regarding the future climate of the habitat. However, detailed background information is required to enable development of plans for protecting and conserving species and for successful assisted migration. Thus, it is important to identify differences between populations in relation to environmental conditions. The genetic diversity of populations, which affect vigor, evolution and adaptability of the species, must also be considered. In this study, we examined 14 populations of P . chihuahuana , with the overall aim of discriminating the populations and form clusters of this species., Methods: Each population was represented by one 50 × 50 m plot established in the center of its respective location. Climate, soil, dasometric, density variables and genetic and species diversities were assessed in these plots for further analyses. The putatively neutral and adaptive AFLP markers were used to calculate genetic diversity. Affinity Propagation (AP) clustering technique and k -means clustering algorithm were used to classify the populations in the optimal number of clusters. Later stepwise binomial logistic regression was applied to test for significant differences in variables of the southern and northern P. chihuahuana populations. Spearman's correlation test was used to analyze the relationships among all variables studied., Results: The binomial logistic regression analysis revealed that seven climate variables, the geographical longitude and sand proportion in the soil separated the southern from northern populations. The northern populations grow in more arid and continental conditions and on soils with lower sand proportion. The mean genetic diversity using all AFLP studied of P. chihuahuana was significantly correlated with the mean temperature in the warmest month, where warmer temperatures are associated to larger genetic diversity. Genetic diversity of P. chihuahuana calculated with putatively adaptive AFLP was not statistically significantly correlated with any environmental factor., Discussion: Future reforestation programs should take into account that at least two different groups (the northern and southern cluster) of P. chihuahuana exist, as local adaptation takes place because of different environmental conditions., Competing Interests: The authors declare there are no competing interests.

Objectives: This study compared two electrode placements ('standard' versus 'nose reference' placement) for measuring oVEMPs, elicited by air-conducted 500Hz tone bursts. The test-retest reliability of both positions was evaluated and additionally both electrode placements were applied on a group of vestibular patients., Methods: Eighteen healthy volunteers (range of 20-25years) participated in the first part and were retested after one week for evaluation of the test-retest reliability. Eleven patients (range of 41-74years) with a variety of vestibular pathologies were tested once., Results: In the normal group, the nose reference electrode placement resulted in significantly larger peak-to-peak amplitudes (p<0.001), shorter n10 (p=0.001) and p15 (p<0.001) latencies and smaller 95% prediction intervals for the Inter-Ocular Ratio (IOR) ([-68, 68] for the standard position versus [-32, 32] for the nose reference position). Furthermore, an excellent amplitude and IOR test-retest reliability was observed with the nose reference configuration, as shown by the intraclass correlation coefficient (ICC), the coefficient of variation of the method error (CV ME ) and the minimal detectable differences (MDD). In the patient group, the same significant amplitude difference was found. Moreover, three patients presented with absent oVEMPs when recorded with the standard placement, whereas the nose reference placement could evoke a detectable oVEMP response., Conclusions: This study demonstrated that a nose reference electrode position results in larger oVEMP amplitudes and achieves a better reliability for the most important clinical parameters (amplitude and IOR). Our patient data substantiate the possible clinical benefit of this position, but further systematic patient verification is required., Significance: The nose reference electrode position facilitates the detection of generally very small oVEMP responses and shows a high test-retest reliability, showing promising potential for future use in the vestibular clinic., (Copyright © 2016 International Federation of Clinical Neurophysiology. Published by Elsevier B.V. All rights reserved.)

Purpose: Adrenocortical carcinoma (ACC) is a rare but aggressive endocrine tumor with limited treatment options. Preclinical studies confirmed overexpression of the chemokine receptor 4 (CXCR4) in this cancer type. This study aimed to analyze the role of CXCR4 imaging using Ga-pentixafor for ACC staging and selection of patients for CXCR4-directed endoradiotherapy., Methods: Thirty patients with histologically proven advanced, metastasized ACC underwent F-FDG PET/CT and Ga-pentixafor PET/CT within a time interval of 3 ± 4 days to evaluate suitability for CXCR4-directed endoradiotherapy. Scans were analyzed retrospectively for visual extent of ACC and SUVmax/mean of the tumor lesions. Ga-pentixafor PET was compared with F-FDG PET, the reference imaging standard. All patients were rated for suitability of CXCR4-directed endoradiotherapy considering patient's history, previous treatment, and CXCR4 expression of FDG-positive lesions compared with background activity within the same organ., Results: All patients had lesions that were positive for both F-FDG and Ga-pentixafor PET and were rated as positive for disease. In 2 patients (7%), Ga-pentixafor PET identified more lesions compared with F-FDG PET. In 5 patients (17%) and 10 patients (33%), complementary and comparable information, respectively, was provided by dual-tracer imaging. In 13 patients (43%), more tumor lesions were identified by F-FDG PET compared with Ga-pentixafor PET. The F-FDG uptake of the malignant lesions was significantly higher (P < 0.01) than the SUVmax/mean for Ga-pentixafor. Overall, 70% of the patients were rated as suitable or potentially suitable for CXCR4-directed treatment., Conclusions: Ga-pentixafor allows in vivo imaging of CXCR4 expression in patients with advanced ACC and may serve as companion diagnostic tool in selecting patients for potential CXCR4-directed endoradiotherapy. Seventy percent of the patients with advanced, metastasized ACC may be suitable for a CXCR4-directed treatment after failure of standard treatment options.

Natural hybrid zones have proven to be precious tools for understanding the origin and maintenance of reproductive isolation (RI) and therefore species. Most available genomic studies of hybrid zones using whole- or partial-genome resequencing approaches have focused on comparisons of the parental source populations involved in genome admixture, rather than exploring fine-scale patterns of chromosomal ancestry across the full admixture gradient present between hybridizing species. We have studied three well-known European 'replicate' hybrid zones of Populus alba and P. tremula, two widespread, ecologically divergent forest trees, using up to 432 505 single-nucleotide polymorphisms (SNPs) from restriction site-associated DNA (RAD) sequencing. Estimates of fine-scale chromosomal ancestry, genomic divergence and differentiation across all 19 poplar chromosomes revealed strikingly contrasting results, including an unexpected preponderance of F1 hybrids in the centre of genomic clines on the one hand, and genomically localized, spatially variable shared variants consistent with ancient introgression between the parental species on the other. Genetic ancestry had a significant effect on survivorship of hybrid seedlings in a common garden trial, pointing to selection against early-generation recombinants. Our results indicate a role for selection against recombinant genotypes in maintaining RI in the face of apparent F1 fertility, consistent with the intragenomic 'coadaptation' model of barriers to introgression upon secondary contact. Whole-genome resequencing of hybridizing populations will clarify the roles of specific genetic pathways in RI between these model forest trees and may reveal which loci are affected most strongly by its cyclic breakdown., (© 2016 John Wiley & Sons Ltd.)

Background: E193, a heterozygous truncating mutation in the human transcription cofactor Eyes absent 4 (Eya4), causes hearing impairment followed by dilative cardiomyopathy., Methods and Results: In this study, we first show Eya4 and E193 alter the expression of p27(kip1) in vitro, suggesting Eya4 is a negative regulator of p27. Next, we generated transgenic mice with cardiac-specific overexpression of Eya4 or E193. Luciferase and chromatin immunoprecipitation assays confirmed Eya4 and E193 bind and regulate p27 expression in a contradictory manner. Activity and phosphorylation status of the downstream molecules casein kinase-2α and histone deacetylase 2 were significantly elevated in Eya4- but significantly reduced in E193-overexpressing animals compared with wild-type littermates. Magnetic resonance imaging and hemodynamic analysis indicate Eya4-overexpression results in an age-dependent development of hypertrophy already under baseline conditions with no obvious functional effects, whereas E193 animals develop onset of dilative cardiomyopathy as seen in human E193 patients. Both cardiac phenotypes were aggravated on pressure overload. Finally, we identified a new heterozygous truncating Eya4 mutation, E215, which leads to similar clinical features of disease and a stable myocardial expression of the mutant protein as seen with E193., Conclusions: Our results implicate Eya4/Six1 regulates normal cardiac function via p27/casein kinase-2α/histone deacetylase 2 and indicate that mutations within this transcriptional complex and signaling cascade lead to the development of cardiomyopathy., (© 2015 American Heart Association, Inc.)

Studying the divergence continuum in plants is relevant to fundamental and applied biology because of the potential to reveal functionally important genetic variation. In this context, whole-genome sequencing (WGS) provides the necessary rigour for uncovering footprints of selection. We resequenced populations of two divergent phylogeographic lineages of Populus alba (n = 48), thoroughly characterized by microsatellites (n = 317), and scanned their genomes for regions of unusually high allelic differentiation and reduced diversity using > 1.7 million single nucleotide polymorphisms (SNPs) from WGS. Results were confirmed by Sanger sequencing. On average, 9134 high-differentiation (≥ 4 standard deviations) outlier SNPs were uncovered between populations, 848 of which were shared by ≥ three replicate comparisons. Annotation revealed that 545 of these were located in 437 predicted genes. Twelve percent of differentiation outlier genome regions exhibited significantly reduced genetic diversity. Gene ontology (GO) searches were successful for 327 high-differentiation genes, and these were enriched for 63 GO terms. Our results provide a snapshot of the roles of 'hard selective sweeps' vs divergent selection of standing genetic variation in distinct postglacial recolonization lineages of P. alba. Thus, this study adds to our understanding of the mechanisms responsible for the origin of functionally relevant variation in temperate trees., (© 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.)

The widespread poplar populations of Sardinia are vegetatively propagated and live in different natural environments forming large monoclonal stands. The main goals of the present study were: i) to investigate/measure the epigenetic diversity of the poplar populations by determining their DNA methylation status; ii) to assess if and how methylation status influences population clustering; iii) to shed light on the changes that occur in the epigenome of ramets of the same poplar clone. To these purposes, 83 white poplar trees were sampled at different locations on the island of Sardinia. Methylation sensitive amplified polymorphism analysis was carried out on the genomic DNA extracted from leaves at the same juvenile stage. The study showed that the genetic biodiversity of poplars is quite limited but it is counterbalanced by epigenetic inter-population molecular variability. The comparison between MspI and HpaII DNA fragmentation profiles revealed that environmental conditions strongly influence hemi-methylation of the inner cytosine. The variable epigenetic status of Sardinian white poplars revealed a decreased number of population clusters. Landscape genetics analyses clearly demonstrated that ramets of the same clone were differentially methylated in relation to their geographic position. Therefore, our data support the notion that studies on plant biodiversity should no longer be restricted to genetic aspects, especially in the case of vegetatively propagated plant species.

We investigate segregation phenomena of particles driven in opposite directions with Brownian dynamics simulations. The particles interact via a repulsive potential and are confined in three-dimensional hard-walled pipes with quadratic cross sections. In a systematic finite-size study, the pipe length is varied. Ordering on finite length scales can be observed, but global segregation seems to vanish for infinite channel length, as it was recently found for lane formation in two dimensions. As an additional effect of the finite hard-walled boundary conditions, interface vibrations and longitudinal demixing are found.

Objective: A systematic literature review and meta-analysis was performed to determine the effect of stimulus type, SCM muscle activation method, transducer type, and method to control SCM muscle EMG level on response parameter values for 0.1-ms click-evoked and 500-Hz tone burst cVEMPs. A description of normative response values was attempted., Design: An electronic systematic literature review was performed to obtain normative cVEMP response data. Subsequently a meta-analysis was conducted to determine significant effects on cVEMP response parameters and to obtain norms., Study Sample: Scopus was used to identify reports containing normative data. Reports were selected based on inclusion and exclusion criteria determined beforehand. Weighted means were calculated and compared to identify significant effects and normative data., Results: Sixty-six reports were included in the systematic review. Stimulus type, SCM muscle activation method, transducer type, and method to control SCM muscle EMG level had significant effects on all response parameters., Conclusions: Optimal stimulus and recording parameters suggested by previous research are confirmed by the current systematic review and meta-analysis and are suggested for clinical use. Response parameter values are influenced by variations in stimulus and recording parameters and normative response values are suggested as guideline for cVEMP interpretation.