The Sanctuary of Despotiko in the Cyclades. Excavations 2001–2012

Auf der heute unbewohnten Insel Despotiko, südwestlich von Antiparos, im Zentrum der Kykladen, liegt an der Stätte Mandra ein Heiligtum. In dessen Zentrum befinden sich das sog. Nord-Temenos und der Südkomplex. Außerhalb davon wurden mindes- tens sechs weitere bauliche Anlagen nachgewiesen. Das von einem Peribolos geschützte Nord-Temenos war der Mittelpunkt der kultischen Aktivitäten. Es setzt sich zusammen aus den Gebäuden A, Δ, Ε, ›Stoen‹ und zwei Zugängen. Im Zentrum liegt ein halbkreis- förmiger Altar. Der Nordteil des Gebäudes A war der ›Tempel‹ des Heiligtums; der Südteil diente vielleicht für Bankette. Die Gebäude Δ und E dienten ebenfalls kultischen Zwecken. Der Südkomplex besteht aus den Bauanlagen Ι and Θ, wobei letztere einen Raum einschließt, der versuchsweise als Kultbad bezeichnet wird. Reiche archäologische Funde bezeugen die Funktion des Heiligtums vom 8. Jh. v. Chr. bis in die hellenistische Zeit – und die Hauptgottheit Apollon. Die baulichen Aktivitäten beschränkten sich auf das 6. und 5. Jh. v. Chr.The sanctuary of Despotiko lies at the ›Mandra‹ site, on an uninhabited island situated to the southwest of Antiparos, in the centre of the Cyclades. The central part comprises the North Temenos and the South complex. Outside at least six other structures were traced. The North Temenos, protected by a peribolos, was the center of the cult activity. It is formed by Buildings A, Δ, Ε, ›stoas‹ and two entrances. In the centre lies a semicircular altar. The North Part of Building A was the ›temple‹ of the sanctuary, the South Part perhaps served for banqueting. Buildings Δ and E served cultic purposes as well. The South Complex consisted of the Building units Ι and Θ, the last including a room tentatively identified as a cultic bath. Rich archaeological finds attest the function of the sanctuary from 8th cent. B.C. to the Hellenistic period – and the main deity, Apollon. The building activity was restricted to the 6th and 5th cent. B.C

Impact of Reduced-Risk Insecticides on Soybean Aphid and Associated Natural Enemies

Insect predators in North America suppress Aphis glycinesMatsumura (Hemiptera: Aphididae) populations; however, insecticides are required when populations reach economically damaging levels. Currently, insecticides used to manage A. glycines are broad-spectrum (pyrethroids and organophosphates), and probably reduce beneficial insect abundance in soybean, Glycine max (L.) Merr. Our goal was to determine whether insecticides considered reduced-risk by the Environmental Protection Agency could protect soybean yield from A. glycines herbivory while having a limited impact on the aphid\u27s natural enemies. We compared three insecticides (imidacloprid, thiamethoxam, and pymetrozine,) to a broad-spectrum insecticide (λ-cyhalothrin) and an untreated control using two application methods. We applied neonicotinoid insecticides to seeds (imidacloprid and thiamethoxam) as well as foliage (imidacloprid); pymetrozine and λ-cyhalothrin were applied only to foliage. Foliage-applied insecticides had lower A. glycines populations and higher yields than the seed-applied insecticides. Among foliage-applied insecticides, pymetrozine and imidacloprid had an intermediate level of A. glycines population and yield protection compared with λ-cyhalothrin and the untreated control. We monitored natural enemies with yellow sticky cards, sweep-nets, and direct observation. Before foliar insecticides were applied (i.e., before aphid populations developed) seed treatments had no observable effect on the abundance of natural enemies. After foliar insecticides were applied, differences in natural enemy abundance were observed when sampled with sweep-nets and direct observation but not with yellow sticky cards. Based on the first two sampling methods, pymetrozine and the foliage-applied imidacloprid had intermediate abundances of natural enemies compared with the untreated control and λ-cyhalothrin

Rapid high-throughput analysis of DNaseI hypersensitive sites using a modified Multiplex Ligation-dependent Probe Amplification approach

BACKGROUND: Mapping DNaseI hypersensitive sites is commonly used to identify regulatory regions in the genome. However, currently available methods are either time consuming and laborious, expensive or require large numbers of cells. We aimed to develop a quick and straightforward method for the analysis of DNaseI hypersensitive sites that overcomes these problems. RESULTS: We have developed a modified Multiplex Ligation-dependent Probe Amplification (MLPA) approach for the identification and analysis of genomic regulatory regions. The utility of this approach was demonstrated by simultaneously analysing 20 loci from the ENCODE project for DNaseI hypersensitivity in a range of different cell lines. We were able to obtain reproducible results with as little as 5 x 10(4) cells per DNaseI treatment. Our results broadly matched those previously reported by the ENCODE project, and both technical and biological replicates showed high correlations, indicating the sensitivity and reproducibility of this method. CONCLUSION: This new method will considerably facilitate the identification and analysis of DNaseI hypersensitive sites. Due to the multiplexing potential of MLPA (up to 50 loci can be examined) it is possible to analyse dozens of DNaseI hypersensitive sites in a single reaction. Furthermore, the high sensitivity of MLPA means that fewer than 10(5) cells per DNaseI treatment can be used, allowing the discovery and analysis of tissue specific regulatory regions without the need for pooling. This method is quick and easy and results can be obtained within 48 hours after harvesting of cells or tissues. As no special equipment is required, this method can be applied by any laboratory interested in the analysis of DNaseI hypersensitive regions

Vacinação compulsória no Brasil: uma reflexão bioética sobre a medida

This article aimed to discuss the compulsory application of vaccines in Brazil in the light of bioethical principles. Historical developments related to the handling of immunizers in the country were analyzed and interferences of the principles proposed by bioethics regarding the safe and effective application of the vaccine were identified. Also, reinforcing the notoriety of the theme in the current scenario, a debate was presented about the new perspectives that the Covid-19 pandemic provided with regard to compulsory vaccination. Finally, the role of the State was examined regarding the balance to be found in the search for the consolidation of the health rights inherent to the social body and the duties of public entities, through the National Immunization Program, in guaranteeing the excellent application of the Citizen Constitution of 1988. The research has a qualitative approach built from a bibliographic review with an  integrative nature. As a conclusion, this study reveals the elementary nature of mandatory vaccination campaigns, especially in scenarios of population uncertainty regarding the safety of immunizers in which the role of government entities is emphasized by the necessary conservation of public health.O presente artigo teve como objetivo discutir a compulsoriedade da aplicação vacinal no Brasil à luz dos princípios bioéticos. Para tanto, analisaram-se os desdobramentos históricos do manuseio dos imunizantes no país e identificaram-se as interferências dos princípios propostos pela bioética no tocante à aplicação segura e eficaz de vacinas. Reforçando-se a notoriedade da temática no cenário atual, debateram-se, ainda, as novas perspectivas que a pandemia de covid-19 propiciou no que concerne à vacinação compulsória. Por último, examinou-se o papel do Estado quanto ao equilíbrio a ser encontrado na busca pela consolidação dos direitos sanitários inerentes ao corpo social e os deveres das entidades públicas, por via do Programa Nacional de Imunizações, na garantia da exímia aplicação da Constituição Cidadã de 1988. A pesquisa teve abordagem qualitativa, construída a partir de revisão bibliográfica de cunho integrativo. Concluiu-se pelo caráter elementar das campanhas de vacinação obrigatória, sobretudo em cenários de incerteza populacional quanto à segurança dos imunizantes, em que o papel dos entes governamentais é enfatizado pela necessária conservação da saúde pública.

Aphids in the air: What is the risk for 2007?

The 2006 growing season has come to a close and with it another year\u27s worth of experience with the soybean aphid. For many, it was a quiet, low aphid year with few reports of economic outbreaks within Iowa. Reflecting these low populations in the field, we observed fewer aphids within our suction traps (Figure 1) compared to the 2005 growing season when the traps collected nearly 100 times more aphids (Figure 2). For the last two years, the Iowa suction trap network (Figure 3) has been part of a larger network of suction traps located in nine states throughout the Midwest

Heterogeneity of Human Neutrophil CD177 Expression Results from CD177P1 Pseudogene Conversion

Most humans harbor both CD177neg and CD177pos neutrophils but 1–10% of people are CD177null, placing them at risk for formation of anti-neutrophil antibodies that can cause transfusion-related acute lung injury and neonatal alloimmune neutropenia. By deep sequencing the CD177 locus, we catalogued CD177 single nucleotide variants and identified a novel stop codon in CD177null individuals arising from a single base substitution in exon 7. This is not a mutation in CD177 itself, rather the CD177null phenotype arises when exon 7 of CD177 is supplied entirely by the CD177 pseudogene (CD177P1), which appears to have resulted from allelic gene conversion. In CD177 expressing individuals the CD177 locus contains both CD177P1 and CD177 sequences. The proportion of CD177hi neutrophils in the blood is a heritable trait. Abundance of CD177hi neutrophils correlates with homozygosity for CD177 reference allele, while heterozygosity for ectopic CD177P1 gene conversion correlates with increased CD177neg neutrophils, in which both CD177P1 partially incorporated allele and paired intact CD177 allele are transcribed. Human neutrophil heterogeneity for CD177 expression arises by ectopic allelic conversion. Resolution of the genetic basis of CD177null phenotype identifies a method for screening for individuals at risk of CD177 isoimmunisation

Small pouches, but high nicotine doses—nicotine delivery and acute effects after use of tobacco-free nicotine pouches

Tobacco-free nicotine pouches are new nicotine products for oral consumption. They can contain very high nicotine amounts that have not been addressed with clinical studies yet. Thus, nicotine delivery, effects on craving, and side effects were assessed using pouches with up to 30 mg nicotine. In this single-center, five-arm, crossover study, 15 regular cigarette smokers consumed tobacco-free nicotine pouches from different brands with 6, 20, and 30 mg for 20 min. Comparators were nicotine-free pouches and tobacco cigarettes. At baseline and predefined time points over a study period of 240 min, plasma nicotine concentrations, effects on cigarette craving, and side effects were assessed. Cardiovascular parameters including arterial stiffness were measured using a MobilOGraph. Consumption of 30 mg nicotine pouches has led to a higher nicotine uptake compared with the cigarette (Cmax: 29.4 vs 15.2 ng/mL; AUC: 45.7 vs 22.1 ng/mL × h). Nicotine uptake in the acute phase was rapid during use of the 30 mg pouch and cigarette. Extraction rate of nicotine differed between pouches. Use of all products has reduced acute cigarette craving, even the nicotine-free pouch. During consumption of the cigarette and the pouches with 20 and 30 mg, heart rate increased about 27, 12, and 25 bpm, respectively. Parameters for arterial stiffness were elevated and all pouches have induced mouth irritations. The pouches with 30 mg nicotine had overall the strongest side effects and may induce addiction. As craving was also reduced by products with less nicotine, it is questionable whether such high nicotine contents should be allowed on the market. A limit of nicotine content is warranted. The nicotine release rate varies across products and needs to be known to estimate the nicotine delivery

Heterogeneity of human Neutrophil CD177 expression results from CD177P1 Pseudogene Conversion

Most humans harbor both CD177neg and CD177pos neutrophils but 1–10% of people are CD177null, placing them at risk for formation of anti-neutrophil antibodies that can cause transfusion-related acute lung injury and neonatal alloimmune neutropenia. By deep sequencing the CD177 locus, we catalogued CD177 single nucleotide variants and identified a novel stop codon in CD177null individuals arising from a single base substitution in exon 7. This is not a mutation in CD177 itself, rather the CD177null phenotype arises when exon 7 of CD177 is supplied entirely by the CD177 pseudogene (CD177P1), which appears to have resulted from allelic gene conversion. In CD177 expressing individuals the CD177 locus contains both CD177P1 and CD177 sequences. The proportion of CD177hi neutrophils in the blood is a heritable trait. Abundance of CD177hi neutrophils correlates with homozygosity for CD177 reference allele, while heterozygosity for ectopic CD177P1 gene conversion correlates with increased CD177neg neutrophils, in which both CD177P1 partially incorporated allele and paired intact CD177 allele are transcribed. Human neutrophil heterogeneity for CD177 expression arises by ectopic allelic conversion. Resolution of the genetic basis of CD177null phenotype identifies a method for screening for individuals at risk of CD177 isoimmunisation

Disorders of sex development: insights from targeted gene sequencing of a large international patient cohort

BACKGROUND: Disorders of sex development (DSD) are congenital conditions in which chromosomal, gonadal, or phenotypic sex is atypical. Clinical management of DSD is often difficult and currently only 13% of patients receive an accurate clinical genetic diagnosis. To address this we have developed a massively parallel sequencing targeted DSD gene panel which allows us to sequence all 64 known diagnostic DSD genes and candidate genes simultaneously. RESULTS: We analyzed DNA from the largest reported international cohort of patients with DSD (278 patients with 46,XY DSD and 48 with 46,XX DSD). Our targeted gene panel compares favorably with other sequencing platforms. We found a total of 28 diagnostic genes that are implicated in DSD, highlighting the genetic spectrum of this disorder. Sequencing revealed 93 previously unreported DSD gene variants. Overall, we identified a likely genetic diagnosis in 43% of patients with 46,XY DSD. In patients with 46,XY disorders of androgen synthesis and action the genetic diagnosis rate reached 60%. Surprisingly, little difference in diagnostic rate was observed between singletons and trios. In many cases our findings are informative as to the likely cause of the DSD, which will facilitate clinical management. CONCLUSIONS: Our massively parallel sequencing targeted DSD gene panel represents an economical means of improving the genetic diagnostic capability for patients affected by DSD. Implementation of this panel in a large cohort of patients has expanded our understanding of the underlying genetic etiology of DSD. The inclusion of research candidate genes also provides an invaluable resource for future identification of novel genes.Stephanie Eggers ... Elizabeth M. Thompson, Jennifer Couper, Anne Baxendale, Jozef Gecz ... et al

Functional characterisation of novel NR5A1 variants reveals multiple complex roles in Disorders of Sex Development

Variants in the NR5A1 gene encoding SF1 have been described in a diverse spectrum of disorders of sex development (DSD). Recently, we reported the use of a targeted gene panel for DSD where we identified 15 individuals with a variant in NR5A1, nine of which are novel. Here, we examine the functional effect of these changes in relation to the patient phenotype. All novel variants tested had reduced trans-activational activity, while several had altered protein level, localization, or conformation. In addition, we found evidence of new roles for SF1 protein domains including a region within the ligand binding domain that appears to contribute to SF1 regulation of Mu¨llerian development. There was little correlation between the severity of the phenotype and the nature of the NR5A1 variant. We report two familial cases of NR5A1 deficiency with evidence of variable expressivity; we also report on individuals with oligogenic inheritance. Finally, we found that the nature of the NR5A1 variant does not inform patient outcomes (including pubertal androgenization and malignancy risk). This study adds nine novel pathogenic NR5A1 variants to the pool of diagnostic variants. It highlights a greater need for understanding the complexity of SF1 function and the additional factors that contribute