Denigrins H-L: Sulfated Derivatives of Denigrins D and E from a New Zealand Dictyodendrilla c.f. dendyi Marine Sponge.

Five new sulfated arylpyrrole and arylpyrrolone alkaloids, denigrins H-L (1-5), along with two known compounds, dictyodendrin B and denigrin G, were isolated from an extract of a New Zealand Dictyodendrilla c.f. dendyi marine sponge. Denigrins H-L represent the first examples of sulfated denigrins, with denigrins H and I (1-2), as derivatives of denigrin D, containing a pyrrolone core, and denigrins J-L (3-5), as derivatives of denigrin E (6), containing a pyrrole core. Their structures were elucidated by interpretation of 1D and 2D NMR spectroscopic data, ESI, and HR-ESI-MS spectrometric data, as well as comparison with literature data. Compounds 1-5, along with six known compounds previously isolated from the same extract, showed minimal cytotoxicity against the HeLa cervical cancer cell line.

Grainyhead-like 2 is required for morphological integrity of mouse embryonic stem cells and orderly formation of inner ear-like organoids.

Mutations in the transcription factor gene grainyhead-like 2 (GRHL2) are associated with progressive non-syndromic sensorineural deafness autosomal dominant type 28 (DFNA28) in humans. Since complete loss of Grhl2 is lethal in mouse embryos, we studied its role during inner ear pathology and hearing loss in vitro. To this end, we generated different homozygous deletions to knockout Grhl2 in mouse embryonic stem cells (Grhl2-KO ESCs), including some mimicking naturally occurring truncations in the dimerisation domain related to human DFNA28. Under naïve culture conditions, Grhl2-KO cells in suspension were more heterogenous in size and larger than wild-type controls. Adherent Grhl2-KO cells were also larger, with a less uniform shape, flattened, less circular morphology, forming loose monolayer colonies with poorly defined edges. These changes correlated with lower expression of epithelial cadherin Cdh1 but no changes in tight junction markers (Ocln, Tjp2) or other Grhl isoforms (Grhl1, Grhl3). Clonogenicity from single cells, proliferation rates of cell populations and proliferation markers were reduced in Grhl2-KO ESCs. We next induced stepwise directed differentiation of Grhl2-KO ESCs along an otic pathway, giving rise to three-dimensional inner ear-like organoids (IELOs). Quantitative morphometry revealed that Grhl2-KO cells initially formed larger IELOs with a less compacted structure, more eccentric shape and increased surface area. These morphological changes persisted for up to one week. They were partially rescued by forced cell aggregation and fully restored by stably overexpressing exogenous Grhl2 in Grhl2-KO ESCs, indicating that Grhl2 alters cell-cell interactions. On day 8, aggregates were transferred into minimal maturation medium to allow self-guided organogenesis for another two weeks. During this period, Grhl2-KO cells and wild-type controls developed similarly, expressing neural, neuronal and sensory hair cell markers, while maintaining their initial differences in size and shape. In summary, Grhl2 is required for morphological maintenance of ESCs and orderly formation of IELOs, consistent with an essential role in organising epithelial integrity during inner ear development. Our findings validate quantitative morphometry as a useful, non-invasive screening method for molecular phenotyping of candidate mutations during organoid development.

Explaining refugee flows. Understanding the 2015 European refugee crisis through a real options lens.

In 2015 the unprecedented arrival of refugees in Europe posed serious challenges for the EU and its member countries on how to deal with such an influx. A key element in better managing refugee flows is to understand what drives these flows in a certain direction. A refugee who comes to Europe has to make trade-offs in terms of cost and benefits, duration, uncertainty and the multi-staged character of the journey. Real options models are a suitable tool for modelling these kind of decision dynamics. On the basis of a case-study, that compares three routes from Syria to Europe, we demonstrate how well the real options analysis is in line with the development of the refugee flows.

Chitosan: A review of molecular structure, bioactivities and interactions with the human body and micro-organisms.

Chitosan has many desirable attributes e.g. antimicrobial properties and promoting wound healing, and is used in various applications. This article first discusses how degree of deacetylation (DD) and molecular weight (MW) impacts on what level of bioactivities chitosan manifests, then introduces the "molecular chain configuration" model to explain various possible mechanisms of antimicrobial interactions between chitosan with different MW and different types of bacteria. Similarly, the possible pathways of how chitosan reacts with cancer and the body's immune system to demonstrate immune and antitumor effects are also discussed by using this model. Moreover, the possible mechanisms of how chitosan enhances coagulation and wound healing are also discussed. With these beneficial bioactivities in mind, the application of chitosan in surgery, tissue engineering and oncology is outlined. This review concludes that as chitosan demonstrates many beneficial bioactivities via multiple mechanisms, it is an important polymer with a promising future in medicine.

Antibacterial Ti-Mn-Cu alloys for biomedical applications.

Titanium alloys are common biomedical materials due to their biocompatibility and mechanical performance. However, titanium alloys are expensive and, unless surface treated, generally cannot prevent surgical infections related to bacteria which can damage the integrity of the implant. In this study, new titanium alloys were developed via powder metallurgy and the addition of manganese and copper, respectively, aiming to limit the manufacturing costs and induce new functionality on the materials including antibacterial response. The addition of manganese and copper to titanium significantly changes the behaviour of the Ti-Mn-Cu alloys leading to the successful stabilization of the beta titanium phase, great refinement of the typical lamellar structure, and achievement of materials with low level of porosity. Consequently, it is found that the mechanical performance and the antibacterial efficacy are enhanced by the addition of a higher amount of alloying elements. The manufactured Ti-Mn-Cu alloys fulfil the requirements for structural biomedical implants and have antibacterial response making them potential candidates for permanent medical implants.

Chitosan: A review of sources and preparation methods.

Chitosan, derived from chitin, has many desirable biomedical attributes. This review aims to explore different sources of chitin and methods of chitosan production with industrial consideration. This article first discussed different sources of chitin for industrial scale production, with considerations given to both their environmental impacts and commercialization potential. Secondly, this article reviews the two categories of chitosan preparation - chemical methods and biological methods - based on existing publications which used lobster by-products as a feedstock source. The mechanisms of the chemical methods are firstly summarized, and then the different chemical agents and reaction parameters used are discussed. Next, both enzymatic and fermentation-based approaches are reviewed under biological methods and compared with chemical methodologies, with lactic fermentation methods as the major focus. This article concludes that lobster cephalothorax could be an ideal source for chitosan preparation on an industrial scale; and chemical methods involve simpler processing overall, while producing chitosan with stronger bioactivities because of the lower molecular weight (MW) and higher degree of deacetylation (DD) achieved by the products. Moreover, due to biological methods inevitably necessitating further chemical processing, an approach involving some unconventional chemical methods has been regarded as a more suitable strategy for industrial scale chitosan production.

Are our beaches safe? Quantifying the human health impact of anthropogenic beach litter on people in New Zealand.

The environmental, social and cultural importance of beaches permeates human society, yet the risk of human injury associated with increasing exposure to anthropogenic beach litter remains an unknown. While the impact of marine debris and beach litter on marine and coastal fauna and flora is a widely reported global issue, we investigate the impact on human health in New Zealand. Anthropogenic beach litter is ubiquitous, few beaches remain pristine, which consequently influences tourist choices and potentially negatively interacts with humans. Human impacts are not well-investigated, with no quantitative studies of impact but many studies qualitatively inferring impact. New Zealand has a socialised medical system allowing a quantitative, decadal assessment of medical insurance claims to determine patterns and trends across ecosystems and causes. We demonstrate for the first time that anthropogenic beach litter poses a common and pervasive exposure hazard to all ages, with specific risk posed to young children. The New Zealand system allows these hazards to be investigated to determine the true effects and costs across a nation, providing an evidence base for decision-makers to address this ubiquitous environmental issue.

Development of a qPCR Method to Measure Mitochondrial and Genomic DNA Damage with Application to Chemotherapy-Induced DNA Damage and Cryopreserved Cells.

DNA damage quantitation assays such as the comet assay have focused on the measurement of total nuclear damage per cell. The adoption of PCR-based techniques to quantify DNA damage has enabled sequence- and organelle-specific assessment of DNA lesions. Here we report on an adaptation of a qPCR technique to assess DNA damage in nuclear and mitochondrial targets relative to control. Novel aspects of this assay include application of the assay to the Rotor-Gene platform with optimized DNA polymerase/fluorophore/primer set combination in a touchdown PCR protocol. Assay validation was performed using ultraviolet C radiation in A549 and THP1 cancer cell lines. A comparison was made to the comet assay applied to peripheral blood mononuclear cells, and an estimation of the effects of cryopreservation on ultraviolet C-induced DNA damage was carried out. Finally, dose responses for DNA damage were measured in peripheral blood mononuclear cells following exposure to the cytotoxic agents bleomycin and cisplatin. We show reproducible experimental outputs across the tested conditions and concordance with published findings with respect to mitochondrial and nuclear genotoxic susceptibilities. The application of this DNA damage assay to a wide range of clinical and laboratory-derived samples is both feasible and resource-efficient.

Selection of oleaginous yeasts for fatty acid production.

BACKGROUND: Oleaginous yeast species are an alternative for the production of lipids or triacylglycerides (TAGs). These yeasts are usually non-pathogenic and able to store TAGs ranging from 20 % to 70 % of their cell mass depending on culture conditions. TAGs originating from oleaginous yeasts can be used as the so-called second generation biofuels, which are based on non-food competing "waste carbon sources". RESULTS: In this study the selection of potentially new interesting oleaginous yeast strains is described. Important selection criteria were: a broad maximum temperature and pH range for growth (robustness of the strain), a broad spectrum of carbon sources that can be metabolized (preferably including C-5 sugars), a high total fatty acid content in combination with a low glycogen content and genetic accessibility. CONCLUSIONS: Based on these selection criteria, among 24 screened species, Schwanniomyces occidentalis (Debaromyces occidentalis) CBS2864 was selected as a promising strain for the production of high amounts of lipids.

Nurses' perceptions of and satisfaction with the use of automated dispensing cabinets at the Heart and Cancer Centers in Qatar: a cross-sectional study.

BACKGROUND: Automated dispensing cabinets (ADCs) were introduced in 2010 and 2012 at the Heart Hospital (HH) and National Center for Cancer Care and Research (NCCCR), both run by Hamad Medical Corporation in Qatar. These medication distribution systems provide computer-controlled storage, dispensing, and tracking of drugs at the point of care in patient care units. The purpose of this study was to assess nurses' perceptions of and satisfaction with the use of ADCs at HH and NCCCR. METHODS: A cross-sectional study was conducted in the two institutions in May and November 2012 using a piloted, validated, online, and anonymous questionnaire. The questionnaire consisted of four parts: nurses' sociodemographic and practice characteristics, 21 questions about their perceptions, one question about their overall satisfaction, and one about the system's ease of use. The self-administered survey was distributed to 503 nurses working at HH and NCCCR over three weeks using Survey Monkey®. RESULTS: The survey response rate was 80 % (n = 403). No significant difference was found in perception scores between the two institutions (p = 0.06). Ninety-four percent (n = 378) of nurses agreed that the medication delivery system allowed them to do their job more safely, and 90 % (n = 363) nurses agreed that they now spent less time waiting for medication from the pharmacy than they did before the ADC system was introduced. Eighty seven percent (n = 349) nurses agreed that they were able to administer medication more efficiently with the ADC system. The overall satisfaction rate (either "very satisfied" or "satisfied") for the two hospitals was 91 %. CONCLUSIONS: The nurses' perceptions of and levels of satisfaction with the ADC system were very good over the 6 months after complete implementation and integration at HH and NCCCR. ADCs appear to increase efficiency in the medication process and should therefore improve the quality of care.

Complete genome sequence of the phenanthrene-degrading soil bacterium Delftia acidovorans Cs1-4.

Polycyclic aromatic hydrocarbons (PAH) are ubiquitous environmental pollutants and microbial biodegradation is an important means of remediation of PAH-contaminated soil. Delftia acidovorans Cs1-4 (formerly Delftia sp. Cs1-4) was isolated by using phenanthrene as the sole carbon source from PAH contaminated soil in Wisconsin. Its full genome sequence was determined to gain insights into a mechanisms underlying biodegradation of PAH. Three genomic libraries were constructed and sequenced: an Illumina GAii shotgun library (916,416,493 reads), a 454 Titanium standard library (770,171 reads) and one paired-end 454 library (average insert size of 8 kb, 508,092 reads). The initial assembly contained 40 contigs in two scaffolds. The 454 Titanium standard data and the 454 paired end data were assembled together and the consensus sequences were computationally shredded into 2 kb overlapping shreds. Illumina sequencing data was assembled, and the consensus sequence was computationally shredded into 1.5 kb overlapping shreds. Gaps between contigs were closed by editing in Consed, by PCR and by Bubble PCR primer walks. A total of 182 additional reactions were needed to close gaps and to raise the quality of the finished sequence. The final assembly is based on 253.3 Mb of 454 draft data (averaging 38.4 X coverage) and 590.2 Mb of Illumina draft data (averaging 89.4 X coverage). The genome of strain Cs1-4 consists of a single circular chromosome of 6,685,842 bp (66.7 %G+C) containing 6,028 predicted genes; 5,931 of these genes were protein-encoding and 4,425 gene products were assigned to a putative function. Genes encoding phenanthrene degradation were localized to a 232 kb genomic island (termed the phn island), which contained near its 3' end a bacteriophage P4-like integrase, an enzyme often associated with chromosomal integration of mobile genetic elements. Other biodegradation pathways reconstructed from the genome sequence included: benzoate (by the acetyl-CoA pathway), styrene, nicotinic acid (by the maleamate pathway) and the pesticides Dicamba and Fenitrothion. Determination of the complete genome sequence of D. acidovorans Cs1-4 has provided new insights the microbial mechanisms of PAH biodegradation that may shape the process in the environment.

Islam, brain death, and transplantation: culture, faith, and jurisprudence.

A significant gap exists between availability of organs for transplant and patients with end-stage organ failure for whom organ transplantation is the last treatment option. Reasons for this mismatch include inadequate approach to potential donor families and donor loss as a result of refractory cardiopulmonary instability during and after brainstem herniation. Other reasons include inadequate cultural competence and sensitivity when communicating with potential donor families. Clinicians may not have an understanding of the cultural and religious perspectives of Muslim families of critically ill patients who may be approached about brain death and organ donation. This review analyzes Islamic cultural and religious perspectives on organ donation, transplantation, and brain death, including faith-based directives from Islamic religious authorities, definitions of death in Islam, and communication strategies when discussing brain death and organ donation with Muslim families. Optimal family care and communication are highlighted using case studies and backgrounds illustrating barriers and approaches with Muslim families in the United States and in the Kingdom of Saudi Arabia that can improve cultural competence and family care as well as increase organ availability within the Muslim population and beyond.

Prevalence of apical periodontitis relative to endodontic treatment in an adult Dutch population: a repeated cross-sectional study.

OBJECTIVE: We aimed to compare an Amsterdam subpopulation's current prevalence of root canal fillings and associated periapical radiolucencies with a similar patient sample from 1988. STUDY DESIGN: An Amsterdam subpopulation was evaluated for missing teeth, restorations, quality of endodontic treatment, and periapical radiolucency. RESULTS: A total of 178 radiographs were evaluated and 4594 teeth were examined. Of these, 324 (7%) exhibited widening of the apical periodontal ligament or periapical radiolucency and 224 (4.8%) had been endodontically treated. A total of 118 teeth (2.5%) had radiographic signs of apical periodontitis. Of these lesions, 54 (45.7%) were linked to endodontically treated teeth (24.1% of endodontically treated teeth). Inadequate root canal fillings were frequent (55.8%). Apical radiolucency was significantly higher in these teeth than in adequately root-filled teeth. CONCLUSION: Findings indicate that the periapical status in an Amsterdam subpopulation has not improved over almost 2 decades.

Mutations in Grxcr1 are the basis for inner ear dysfunction in the pirouette mouse.

Recessive mutations at the mouse pirouette (pi) locus result in hearing loss and vestibular dysfunction due to neuroepithelial defects in the inner ear. Using a positional cloning strategy, we have identified mutations in the gene Grxcr1 (glutaredoxin cysteine-rich 1) in five independent allelic strains of pirouette mice. We also provide sequence data of GRXCR1 from humans with profound hearing loss suggesting that pirouette is a model for studying the mechanism of nonsyndromic deafness DFNB25. Grxcr1 encodes a 290 amino acid protein that contains a region of similarity to glutaredoxin proteins and a cysteine-rich region at its C terminus. Grxcr1 is expressed in sensory epithelia of the inner ear, and its encoded protein is localized along the length of stereocilia, the actin-filament-rich mechanosensory structures at the apical surface of auditory and vestibular hair cells. The precise architecture of hair cell stereocilia is essential for normal hearing. Loss of function of Grxcr1 in homozygous pirouette mice results in abnormally thin and slightly shortened stereocilia. When overexpressed in transfected cells, GRXCR1 localizes along the length of actin-filament-rich structures at the dorsal-apical surface and induces structures with greater actin filament content and/or increased lengths in a subset of cells. Our results suggest that deafness in pirouette mutants is associated with loss of GRXCR1 function in modulating actin cytoskeletal architecture in the developing stereocilia of sensory hair cells.

Ferroelectric thin-film capacitors and piezoelectric switches for mobile communication applications.

Thin-film ferroelectric capacitors have been integrated with resistors and active functions such as ESD protection into small, miniaturized modules, which enable a board space saving of up to 80%. With the optimum materials and processes, integrated capacitors with capacitance densities of up to 100 nF/mm2 for stacked capacitors combined with breakdown voltages of 90 V have been achieved. The integration of these high-density capacitors with extremely high breakdown voltage is a major accomplishment in the world of passive components and has not yet been reported for any other passive integration technology. Furthermore, thin-film tunable capacitors based on barium strontium titanate with high tuning range and high quality factor at 1 GHz have been demonstrated. Finally, piezoelectric thin films for piezoelectric switches with high switching speed have been realized.

Integrated ferroelectric stacked MIM capacitors with 100 nF/mm(2) and 90 V breakdown as replacement for discretes.

This paper shows for the first time integrated thin film ferroelectric metal-insulator-metal capacitors on silicon with a record high capacitance density above 100 nF/mm(2) combined with a breakdown voltage of 90 V and a lifetime exceeding 10 years at 85 degrees C and 5 V. The high capacitance density was obtained by a combination of material optimizations resulting in a dielectric constant of 1600, and stacking of capacitors. The reliability of these ferroelectric capacitors was studied in detail with accelerated lifetime testing. The high performance of the integrated capacitors in this paper shows great potential for applications demanding high capacitance densities combined with electrostatic discharge protection.

Signatures from tissue-specific MPSS libraries identify transcripts preferentially expressed in the mouse inner ear.

Specialization in cell function and morphology is influenced by the differential expression of mRNAs, many of which are expressed at low abundance and restricted to certain cell types. Detecting such transcripts in cDNA libraries may require sequencing millions of clones. Massively parallel signature sequencing (MPSS) is well suited to identifying transcripts that are expressed in discrete cell types and in low abundance. We have made MPSS libraries from microdissections of three inner ear tissues. By comparing these MPSS libraries to those of 87 other tissues included in the Mouse Reference Transcriptome online resource, we have identified genes that are highly enriched in, or specific to, the inner ear. We show by RT-PCR and in situ hybridization that signatures unique to the inner ear libraries identify transcripts with highly specific cell-type localizations. These transcripts serve to illustrate the utility of a resource that is available to the research community. Utilization of these resources will increase the number of known transcription units and expand our knowledge of the tissue-specific regulation of the transcriptome.

Mutation of a transcription factor, TFCP2L3, causes progressive autosomal dominant hearing loss, DFNA28.

We ascertained a large American family with an autosomal dominant form of progressive non-syndromic sensorineural hearing loss. After excluding linkage to known deafness loci, we performed a genome-wide scan and found linkage to marker GAAT1A4 on chromosome 8q22 (LOD=5.12 at theta=0), and this locus was designated DFNA28. Sequencing of six candidate genes in the 1.4 cM linked region identified a frameshift mutation (1609-1610insC) resulting in a premature translation stop codon in exon 14 of the gene TFCP2L3 (transcription factor cellular promoter 2-like 3). TFCP2L3 is a member of a family of transcription factor genes whose archetype is TFCP2, a mammalian homolog of the Drosophila gene grainyhead. Northern blot analyses and in situ hybridization studies show that mouse Tfcp2l3 is expressed in many epithelial tissues, including cells lining the cochlear duct, at embryonic day 18.5 and postnatal day 5.

Dominant and recessive deafness caused by mutations of a novel gene, TMC1, required for cochlear hair-cell function.

Positional cloning of hereditary deafness genes is a direct approach to identify molecules and mechanisms underlying auditory function. Here we report a locus for dominant deafness, DFNA36, which maps to human chromosome 9q13-21 in a region overlapping the DFNB7/B11 locus for recessive deafness. We identified eight mutations in a new gene, transmembrane cochlear-expressed gene 1 (TMC1), in a DFNA36 family and eleven DFNB7/B11 families. We detected a 1.6-kb genomic deletion encompassing exon 14 of Tmc1 in the recessive deafness (dn) mouse mutant, which lacks auditory responses and has hair-cell degeneration. TMC1 and TMC2 on chromosome 20p13 are members of a gene family predicted to encode transmembrane proteins. Tmc1 mRNA is expressed in hair cells of the postnatal mouse cochlea and vestibular end organs and is required for normal function of cochlear hair cells.