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Antihydrogen atoms, H―, are now routinely created and can be stored for long enough to allow comparison with ordinary matter. A major goal of these efforts is to test potential physics beyond the Standard Model. This will require further developments in the experiments including the accumulation of more antihydrogen atoms and their storage over longer times. The latter is limited by the unavoidable presence of normal hydrogen molecules. Interactions of H― with H2 lead to the destruction of the hard-won antimatter. Little is known about these interactions but quantitative information will be crucial in guiding experimental developments. Physically realistic modelling of rearrangement scattering of “heavy” antimatter particles (antiprotons, p―, and antihydrogen atoms) by normal matter molecules, such as H2 + H―→ Pn + Ps + H (where Pn represents protonium and Ps positronium), requires the development of new theoretical and computational methodologies. R-matrix theory offers a strong prospect for tackling such problems having proved itself in atomic, molecular and optical physics. It divides the problem into a computationally demanding but energy-independent inner region and simpler energy-dependent outer regions. We propose to adapt the new RmatReact ultracold chemistry approach for the more complex molecular matter-antimatter problems. Here, developments required for the inner region, the boundary and outer regions are outlined. We also report some preliminary bound-state calculations on the {p,p,p―} system and a study of the required mixed coordinate systems for the general effective 3-body case and their transformations at the R-matrix boundary surfaces.

Theory and model evidence indicate a higher global hydrological sensitivity for the same amount of surface warming to solar as to greenhouse gas (GHG) forcing, but regional patterns are highly uncertain due to their dependence on circulation and dynamics. We analyse a multi-model ensemble of idealized experiments and a set of simulations of the last millennium and we demonstrate similar global signatures and patterns of forced response in the tropical Pacific, of higher sensitivity for the solar forcing. In the idealized simulations, both solar and GHG forcing warm the equatorial Pacific, enhance precipitation in the central Pacific, and weaken and shift the Walker circulation eastward. Centennial variations in the solar forcing over the last millennium cause similar patterns of enhanced equatorial precipitation and slowdown of the Walker circulation in response to periods with stronger solar forcing. Similar forced patterns albeit of considerably weaker magnitude are identified for variations in GHG concentrations over the 20th century, with the lower sensitivity explained by fast atmospheric adjustments. These findings differ from previous studies that have typically suggested divergent responses in tropical precipitation and circulation between the solar and GHG forcings. We conclude that tropical Walker circulation and precipitation might be more susceptible to solar variability rather than GHG variations during the last-millennium, assuming comparable global mean surface temperature changes.

Abstract Background Genome-wide association studies have been successful in finding common variants influencing common traits. However, these associations only account for a fraction of trait heritability. There has been a shift in the field towards studying low frequency and rare variants, which are now widely recognised as putative complex trait determinants. Despite this increasing focus on examining the role of low frequency and rare variants in complex disease susceptibility, there is a lack of user-friendly analytical packages implementing powerful association tests for the analysis of rare variants. Results We have developed two software tools, CCRaVAT (Case-Control Rare Variant Analysis Tool) and QuTie (Quantitative Trait), which enable efficient large-scale analysis of low frequency and rare variants. Both programs implement a collapsing method examining the accumulation of low frequency and rare variants across a locus of interest that has more power than single variant analysis. CCRaVAT carries out case-control analyses whereas QuTie has been developed for continuous trait analysis. Conclusions CCRaVAT and QuTie are easy to use software tools that allow users to perform genome-wide association analysis on low frequency and rare variants for both binary and quantitative traits. The software is freely available and provides the genetics community with a resource to perform association analysis on rarer genetic variants.

Abstract Background The number of women working in general practice internationally has been steadily rising. In Scotland there have been concerns that such a change may lead to increased part-time working and subsequently to a fall in available general practice manpower despite an apparently rising overall number of general practitioners. However, there is very little information on the actual hours worked by men and women general practitioners or on the types of work they are undertaking. Methods Anonymous workload questionnaires of all Scottish general practitioner principals and non-principals Results Response rates for general practice principals and non-principals were 67.2% and 65.2% respectively. Male principals spent on average 18% more time on general medical services (GMS) and 50% more time on non-GMS activities (such as teaching, specialist sessions, administration and research) than women (both p Conclusion Women doctors in primary care in Scotland work fewer hours in all age groups than their male counterparts. The rapidly increasing proportion of women in general practice may lead to an increasing shortfall of medical availability in the future if current work patterns are maintained. Further longitudinal research is required to establish this and man-power planning is required now to address this. More worryingly auxiliary activities such as teaching and administrative duties are not being taken up by women. This may have serious implications for the future development of the specialty in Scotland.

Objectives: Burnout and occupational stress are frequently experienced by healthcare professionals (HCPs). Mindfulness-based stress reduction (MBSR) has been found to improve the psychological health outcomes of HCPs. To date, systematic reviews and meta-analyses have primarily focused upon empirical investigations into the reduction of stress amongst HCPs using MBSR and are limited to empirical studies published before December 2019. This systematic review aimed to update the current evidence base and broaden our understanding of the effectiveness of MBSR on improving the psychological functioning of HCPs., Methods: Three electronic databases (Medline, Psych Info and Web of Science) were searched without time frame restrictions. Quantitative studies included randomised controlled trials, clinical controlled trials, pre-post designs and studies with up to a 12-month follow-up period. All studies included in the review employed a MBSR programme, standardised measures of psychological functioning and qualified HCPs as participants., Results: Using PRISMA guidelines thirty studies were included in the review. The reviewed literature suggested that MBSR was effective in reducing HCPs experiences of anxiety, depression and stress. MBSR was also found to be effective in increasing HCP levels of mindfulness and self-compassion. However, MBSR did not appear as effective in reducing burnout or improving resilience amongst HCPs. Abbreviated MBSR programmes were found to be as effective as the traditional 8-week MBSR programmes., Conclusions: MBSR is an effective intervention which can help improve the psychological functioning of HCPs. Recommendations include improving the overall quality of the studies by employing more robust controlled designs with randomisation, increased sample sizes with heterogeneous samples, and making active comparisons between interventions used., Competing Interests: Conflict of InterestThe authors declare they have no conflict of interest., (© Springer Science+Business Media, LLC, part of Springer Nature 2020.)

Invasion by generalist tree species can cause biotic homogenization, and such community impoverishment is likely more important in rare forest types. We quantified changes in tree diversity within Carolinian (range in Central Hardwood Forest), central (range in Central Hardwood Forest and Northern Hardwood-Conifer Forest), and northern species [range reached Northern-Conifer-Hardwood/closed Boreal (spruce-Fir) Forest] in an old forest tract in southern Canada at points surveyed 24 years apart. We asked: How did mature tree and sapling composition and abundance change for the three species' groups? Did those changes lead to biotic homogenization? Can species' changes be explained by community traits? We tested for differences in temporal and spatial tree β-diversity, as well as forest composition and structure, using univariate/multivariate analyses and a community trait-based approach to identify drivers of change. Major increases occurred in abundance for mature Acer rubrum (northern), while other species decreased ( Fraxinus americana , Populus grandidentata ); declines were found in A. saccharinum (central) and Cornus florida (Carolinian). Species composition of saplings, but not mature trees, changed due to replacement; no evidence for biotic homogenization existed in either cohort. As a group, northern mature tree species increased significantly, while central species decreased; saplings of pooled Carolinian species also declined. Shade tolerance in mature trees increased, reflecting successional changes, while drought tolerance decreased, perhaps due to changing temperatures, altered precipitation or ground water levels. Saplings showed declines in all traits, probably because of compositional change. Our results demonstrated that saplings can more closely reflect change in forest dynamics than mature trees, especially over short time periods. Based on sapling trends, this remnant could ultimately transition to a mesophytic hardwood stand dominated by A. rubrum and other shade-tolerant species, creating a more homogeneous forest. While encouraging regeneration for Carolinian and central tree species could ensure high levels of diversity are conserved in the future, it is important to balance this with the primary management goal of maintaining the forest's old-growth characteristics., Competing Interests: None declared., (© 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Rationale: Idiopathic or heritable pulmonary arterial hypertension is characterized by loss and obliteration of lung vasculature. Endothelial cell dysfunction is pivotal to the pathophysiology, but different causal mechanisms may reflect a need for patient-tailored therapies., Objectives: Endothelial cells differentiated from induced pluripotent stem cells were compared with pulmonary arterial endothelial cells from the same patients with idiopathic or heritable pulmonary arterial hypertension, to determine whether they shared functional abnormalities and altered gene expression patterns that differed from those in unused donor cells. We then investigated whether endothelial cells differentiated from pluripotent cells could serve as surrogates to test emerging therapies., Methods: Functional changes assessed included adhesion, migration, tube formation, and propensity to apoptosis. Expression of bone morphogenetic protein receptor type 2 (BMPR2) and its target, collagen IV, signaling of the phosphorylated form of the mothers against decapentaplegic proteins (pSMAD1/5), and transcriptomic profiles were also analyzed., Measurements and Main Results: Native pulmonary arterial and induced pluripotent stem cell-derived endothelial cells from patients with idiopathic and heritable pulmonary arterial hypertension compared with control subjects showed a similar reduction in adhesion, migration, survival, and tube formation, and decreased BMPR2 and downstream signaling and collagen IV expression. Transcriptomic profiling revealed high kisspeptin 1 (KISS1) related to reduced migration and low carboxylesterase 1 (CES1), to impaired survival in patient cells. A beneficial angiogenic response to potential therapies, FK506 and Elafin, was related to reduced slit guidance ligand 3 (SLIT3), an antimigratory factor., Conclusions: Despite the site of disease in the lung, our study indicates that induced pluripotent stem cell-derived endothelial cells are useful surrogates to uncover novel features related to disease mechanisms and to better match patients to therapies.

The polyamine spermidine is absolutely required for growth and cell proliferation in eukaryotes, due to its role in post-translational modification of essential translation elongation factor eIF5A, mediated by deoxyhypusine synthase. We have found that free-living ciliates Tetrahymena and Paramecium lost the eukaryotic genes encoding spermidine biosynthesis: S-adenosylmethionine decarboxylase (AdoMetDC) and spermidine synthase (SpdSyn). In Tetrahymena, they were replaced by a gene encoding a fusion protein of bacterial AdoMetDC and SpdSyn, present as three copies. In Paramecium, a bacterial homospermidine synthase replaced the eukaryotic genes. Individual AdoMetDC-SpdSyn fusion protein paralogues from Tetrahymena exhibit undetectable AdoMetDC activity; however, when two paralogous fusion proteins are mixed, AdoMetDC activity is restored and spermidine is synthesized. Structural modelling indicates a functional active site is reconstituted by sharing critical residues from two defective protomers across the heteromer interface. Paramecium was found to accumulate homospermidine, suggesting it replaces spermidine for growth. To test this concept, a budding yeast spermidine auxotrophic strain was found to grow almost normally with homospermidine instead of spermidine. Biosynthesis of spermidine analogue aminopropylcadaverine, but not exogenously provided norspermidine, correlated with some growth. Finally, we found that diverse single-celled eukaryotic parasites and multicellular metazoan Schistosoma worms have lost the spermidine biosynthetic pathway but retain deoxyhypusine synthase., (© 2015 John Wiley & Sons Ltd.)

This unit describes a procedure for generating human induced pluripotent stem cells (hiPSCs) using the Laser-Enabled Analysis and Processing (LEAP®) system, which combines high-throughput cell imaging with laser-mediated cell manipulation. Use of this system should not only improve the quality and uniformity of hiPSCs produced, but ultimately enable a more rapid, efficient, high-throughput, and automated production process., (Copyright © 2014 John Wiley & Sons, Inc.)

Structural backbones of iron-scavenging siderophore molecules include polyamines 1,3-diaminopropane and 1,5-diaminopentane (cadaverine). For the cadaverine-based desferroxiamine E siderophore in Streptomyces coelicolor, the corresponding biosynthetic gene cluster contains an ORF encoded by desA that was suspected of producing the cadaverine (decarboxylated lysine) backbone. However, desA encodes an l-2,4-diaminobutyrate decarboxylase (DABA DC) homologue and not any known form of lysine decarboxylase (LDC). The only known function of DABA DC is, together with l-2,4-aminobutyrate aminotransferase (DABA AT), to synthesize 1,3-diaminopropane. We show here that S. coelicolor desA encodes a novel LDC and we hypothesized that DABA DC homologues present in siderophore biosynthetic clusters in the absence of DABA AT ORFs would be novel LDCs. We confirmed this by correctly predicting the LDC activity of a DABA DC homologue from a Yersinia pestis siderophore biosynthetic pathway. The corollary was confirmed for a DABA DC homologue, adjacent to a DABA AT ORF in a siderophore pathway in the cyanobacterium Anabaena variabilis, which was shown to be a bona fide DABA DC. These findings enable prediction of whether a siderophore pathway will utilize 1,3-diaminopropane or cadaverine, and suggest that the majority of bacteria use DABA AT and DABA DC for siderophore, rather than norspermidine/polyamine biosynthesis., (© 2012 Blackwell Publishing Ltd.)

During culture adaptation, human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) tend to acquire chromosomal aberrations. Generally, stem cell lines are screened for large-scale chromosomal changes using low resolution karyotype analysis. Recent studies characterizing human stem cells using array-comparative genomic hybridization (aCGH) suggests most abnormalities acquired during culture are under the resolution of karyotype analysis and therefore are routinely missed. Here, we describe a custom-designed stem cell focused microarray utilizing 44K probes, with increased resolution in relevant stem cell-associated and cancer-related genes.

Proper maintenance of stem cells is essential for successful utilization of ESCs/iPSCs as tools in developmental and drug discovery studies and in regenerative medicine. Standardization is critical for all future applications of stem cells and necessary to fully understand their potential. This study reports a novel approach for the efficient, consistent expansion of human ESCs and iPSCs using laser sectioning, instead of mechanical devices or enzymes, to divide cultures into defined size clumps for propagation. Laser-mediated propagation maintained the pluripotency, quality, and genetic stability of ESCs/iPSCs and led to enhanced differentiation potential. This approach removes the variability associated with ESC/iPSC propagation, significantly reduces the expertise, labor, and time associated with manual passaging techniques and provides the basis for scalable delivery of standardized ESC/iPSC lines. Adoption of standardized protocols would allow researchers to understand the role of genetics, environment, and/or procedural effects on stem cells and would ensure reproducible production of stem cell cultures for use in clinical/therapeutic applications.

Culturing stem cells for an extended period of time can lead to acquired chromosomal aberrations. Determining the copy number variant (CNV) profile of stem cell lines is critical since CNVs can have dramatic effects on gene expression and tumorigenic potential. Here, we describe an improved version of our StemArray, a stem-cell-focused comparative genomic hybridization (aCGH) microarray, which contains 135,000 probes and covers over 270 stem cell and cancer related genes at the exon level. We have dramatically increased the median probe spacing throughout the genome in order to obtain a higher resolution genetic profile of the cell lines. To illustrate the importance of using the StemArray, we describe a karyotypically normal iPSC line in which we detected acquired chromosomal variations that could affect the cellular phenotype of the cells. Identifying adaptive chromosomal aberrations in stem cell lines is essential if they are to be used in regenerative medicine.

We have identified gene fusions of polyamine biosynthetic enzymes S-adenosylmethionine decarboxylase (AdoMetDC, speD) and aminopropyltransferase (speE) orthologues in diverse bacterial phyla. Both domains are functionally active and we demonstrate the novel de novo synthesis of the triamine spermidine from the diamine putrescine by fusion enzymes from β-proteobacterium Delftia acidovorans and δ-proteobacterium Syntrophus aciditrophicus, in a ΔspeDE gene deletion strain of Salmonella enterica sv. Typhimurium. Fusion proteins from marine α-proteobacterium Candidatus Pelagibacter ubique, actinobacterium Nocardia farcinica, chlorobi species Chloroherpeton thalassium, and β-proteobacterium D. acidovorans each produce a different profile of non-native polyamines including sym-norspermidine when expressed in Escherichia coli. The different aminopropyltransferase activities together with phylogenetic analysis confirm independent evolutionary origins for some fusions. Comparative genomic analysis strongly indicates that gene fusions arose by merger of adjacent open reading frames. Independent fusion events, and horizontal and vertical gene transfer contributed to the scattered phyletic distribution of the gene fusions. Surprisingly, expression of fusion genes in E. coli and S. Typhimurium revealed novel latent spermidine catabolic activity producing non-native 1,3-diaminopropane in these species. We have also identified fusions of polyamine biosynthetic enzymes agmatine deiminase and N-carbamoylputrescine amidohydrolase in archaea, and of S-adenosylmethionine decarboxylase and ornithine decarboxylase in the single-celled green alga Micromonas., (© 2011 Blackwell Publishing Ltd.)

The ability to realize the full potential of human pluripotent stem cells (hPSCs) as tools for -understanding human development and advancing the field of regenerative medicine is dependent on efficient methods to genetically manipulate these cells. There are several methods for introducing foreign DNA into cells such as electroporation, lipid-based transfection technology, and viral transduction. We describe here a method to transfect human embryonic stem cells (hESCs) using nucleofection technology. This unique method uses the Nucleofector II Device that combines the use of a cell type-specific Nucleofector Solution and preprogrammed electrical parameters to efficiently deliver DNA into the cell nucleus. The use of this technology allows high-efficiency transfer of nucleic acids into hESCs enabling both transient and stable manipulation of gene expression in these cells.

Human embryonic and induced pluripotent stem cells (ESCs, iPSCs) that are cultured for an extended period of time are susceptible to genomic instability. Chromosomal aberrations decrease the reliability and reproducibility of experiments and could deem the cells unusable for therapeutic purposes. The genetic stability of human ESCs and iPSCs is commonly monitored by karyotype analysis. However, this low-resolution technique can only identify large aneuploidies. A reliable, high-resolution technique to detect genomic aberrations at a cost comparable to karyotyping is needed to better characterize stem cell lines. We have designed a stem cell focused array-comparative genomic hybridization microarray that covers the entire genome at high resolution with increased probe coverage in over 60 stem cell associated genes and more than 195 cancer related genes. Several iPSC lines were analyzed using the focused microarray and compared with either karyotyping or a standard Agilent 44K microarray. In addition to the abnormalities detected by these platforms, the custom microarray identified several small duplications spanning stem cell and/or cancer related genes. Scientists using a stem cell focused microarray to characterize their stem cells will be aware of the structural variants present in their cells and be more confident in their experimental results.

Polyamine biosynthesis in plants differs from other eukaryotes because of the contribution of genes from the cyanobacterial ancestor of the chloroplast. Plants possess an additional biosynthetic route for putrescine formation from arginine, consisting of the enzymes arginine decarboxylase, agmatine iminohydrolase and N-carbamoylputrescine amidohydrolase, derived from the cyanobacterial ancestor. They also synthesize an unusual tetraamine, thermospermine, that has important developmental roles and which is evolutionarily more ancient than spermine in plants and algae. Single-celled green algae have lost the arginine route and are dependent, like other eukaryotes, on putrescine biosynthesis from the ornithine. Some plants like Arabidopsis thaliana and the moss Physcomitrella patens have lost ornithine decarboxylase and are thus dependent on the arginine route. With its dependence on the arginine route, and the pivotal role of thermospermine in growth and development, Arabidopsis represents the most specifically plant mode of polyamine biosynthesis amongst eukaryotes. A number of plants and algae are also able to synthesize unusual polyamines such as norspermidine, norspermine and longer polyamines, and biosynthesis of these amines likely depends on novel aminopropyltransferases similar to thermospermine synthase, with relaxed substrate specificity. Plants have a rich repertoire of polyamine-based secondary metabolites, including alkaloids and hydroxycinnamic amides, and a number of polyamine-acylating enzymes have been recently characterised. With the genetic tools available for Arabidopsis and other model plants and algae, and the increasing capabilities of comparative genomics, the biological roles of polyamines can now be addressed across the plant evolutionary lineage., (2010 Elsevier Masson SAS. All rights reserved.)

Polyamines are small flexible organic polycations found in almost all cells. They likely existed in the last universal common ancestor of all extant life, and yet relatively little is understood about their biological function, especially in bacteria and archaea. Unlike eukaryotes, where the predominant polyamine is spermidine, bacteria may contain instead an alternative polyamine, sym-homospermidine. We demonstrate that homospermidine synthase (HSS) has evolved vertically, primarily in the alpha-Proteobacteria, but enzymatically active, diverse HSS orthologues have spread by horizontal gene transfer to other bacteria, bacteriophage, archaea, eukaryotes, and viruses. By expressing diverse HSS orthologues in Escherichia coli, we demonstrate in vivo the production of co-products diaminopropane and N(1)-aminobutylcadaverine, in addition to sym-homospermidine. We show that sym-homospermidine is required for normal growth of the alpha-proteobacterium Rhizobium leguminosarum. However, sym-homospermidine can be replaced, for growth restoration, by the structural analogues spermidine and sym-norspermidine, suggesting that the symmetrical or unsymmetrical form and carbon backbone length are not critical for polyamine function in growth. We found that the HSS enzyme evolved from the alternative spermidine biosynthetic pathway enzyme carboxyspermidine dehydrogenase. The structure of HSS is related to lysine metabolic enzymes, and HSS and carboxyspermidine dehydrogenase evolved from the aspartate family of pathways. Finally, we show that other bacterial phyla such as Cyanobacteria and some alpha-Proteobacteria synthesize sym-homospermidine by an HSS-independent pathway, very probably based on deoxyhypusine synthase orthologues, similar to the alternative homospermidine synthase found in some plants. Thus, bacteria can contain alternative biosynthetic pathways for both spermidine and sym-norspermidine and distinct alternative pathways for sym-homospermidine.

Background: Telomerase expression is detectable in 81-95% of breast carcinomas and may serve as a therapeutic target. The objective of this study was to investigate repression of telomerase activity in primary ductal breast cancer cells through transcriptional regulation of the catalytic subunit hTERT. We hypothesized that inhibition of telomerase expression could be achieved via Tat mediated protein transduction of the repressor protein E2F-1., Methods: Protein purification techniques were refined to yield biologically active Tat fusion proteins (TFPs) capable of transducing the breast cancer cell lines HCC1937 and HCC1599. Cell lines were treated with wildtype E2F-1 (E2F-1/TatHA), mutant E2F-1 (E132/TatHA) and a control Tat peptide (TatHA) for 24 hours. Total RNA was isolated from treated cells, reverse transcribed and fold changes in gene expression for hTERT determined via real-time RT-qPCR., Results: Significant repression of the catalytic subunit of telomerase (hTERT) was present in both HCC1937 and HCC1599 cells following treatment with E2F-1/TatHA. In HCC1937 cells, hTERT was repressed 3.5-fold by E2F-1/TatHA in comparison to E132/TatHA (p < 0.0012) and the TatHA peptide controls (p < 0.0024). In HCC1599 cells, hTERT was also repressed with E2F-1/TatHA treatment by 4.0-fold when compared to the E132/TatHA control (p < 0.0001). A slightly lower hTERT repression of 3.3-fold was observed with E2F-1/TatHA in the HCC1599 cells when compared to the TatHA control (p < 0.0001)., Conclusion: These results suggest that transduction of E2F-1/TatHA fusion proteins in vitro is an effective repressor of hTERT expression in the primary ductal breast cancer cell lines HCC1937 and HCC1599.

Polygalacturonases (PGs) have been proposed to play an important role in the process of cell separation. The Arabidopsis thaliana genome contains 69 annotated genes that by amino acid homology and transcript organization could be classified as putative PGs and these can be grouped into multiple clades. An analysis of five members located in two separate clades, using reporter fusion constructs and reverse transcription-PCR, revealed that whilst these PGs exhibit high sequence similarity they have distinct patterns of spatial and temporal expression. Sites of expression include the aleurone and endosperm cells surrounding the emerging radicle in a germinating seed, the cortical cells adjacent to the developing lateral root, the abscission zones of floral organs, the dehiscence zone of anthers and siliques, and pollen grains. Silencing of an abscission-related PG (At2g41850), using a T-DNA insertion strategy, delayed the time-course of floral organ loss but did not prevent shedding from taking place. These observations are discussed with regard to the contribution that PGs may play during the life cycle of a plant.

A fundamental issue in the safety assessment of genetically modified crops is the question of whether unintentional changes have occurred in the crop plant as a consequence of the genetic modification. This question was addressed recently by using a powerful metabolite fingerprinting and metabolite profiling method to assess whether genetically modified potatoes are substantially similar to their corresponding conventional cultivars.

Background: Chronic rhinosinusitis (CRS) refractory to medical and surgical therapy is a difficult problem for patients and physicians. Topical antimicrobial nasal irrigations are commonly used for treatment with great variation in methodology and without clear scientific support for current treatment formulations. The purpose of this study was to develop a scientific rationale for creating standardized recommendations for clinical practice in the use of topical antimicrobial washes for CRS., Methods: An extensive review of basic science and clinical literature on the treatment of CRS with topical antimicrobial washes was completed. Pharmacokinetics of and organism susceptibility to appropriate topically applied antimicrobial agents were reviewed., Results: The most common organisms associated with CRS were identified. The relevant pharmacokinetics of drugs targeted at these organisms are presented. Susceptibility breakpoints set by the National Committee for Clinical Laboratory Standards are identified to help establish the most effective concentration of the identified drugs. Recommendations for agent selection, agent concentration, length of treatment, dosing schedule, and methods of irrigation are presented., Conclusion: Antimicrobial nasal washes provide a potentially effective treatment for the growing population of patients who remain symptomatic after appropriate medical and surgical intervention. This study establishes the basic principles supporting this treatment option and offers rational, evidence-based treatment guidelines. The study has identified additional areas that need to be investigated before prospective clinical trials can be effectively undertaken.

A novel form of translational regulation is described for the key polyamine biosynthetic enzyme S-adenosylmethionine decarboxylase (AdoMetDC). Plant AdoMetDC mRNA 5' leaders contain two highly conserved overlapping upstream open reading frames (uORFs): the 5' tiny and 3' small uORFs. We demonstrate that the small uORF-encoded peptide is responsible for constitutively repressing downstream translation of the AdoMetDC proenzyme ORF in the absence of increased polyamine levels. This first example of a sequence-dependent uORF to be described in plants is also functional in Saccharomyces cerevisiae. The tiny uORF is required for normal polyamine-responsive AdoMetDC mRNA translation, and we propose that this is achieved by control of ribosomal recognition of the occluded small uORF, either by ribosomal leaky scanning or by programmed -1 frameshifting. In vitro expression demonstrated that both the tiny and the small uORFs are translated. This tiny/small uORF configuration is highly conserved from moss to Arabidopsis thaliana, and a more diverged tiny/small uORF arrangement is found in the AdoMetDC mRNA 5' leader of the single-celled green alga Chlamydomonas reinhardtii, indicating an ancient origin for the uORFs.

Objectives/hypothesis: Nasopalatine duct cysts are the most common cystic lesion of nonodontogenic origin of the maxilla. The purposes of the study were to review the epidemiology and clinical presentation, to describe the radiographic and pathological findings, and to discuss surgical management of this entity., Study Design: Case presentations of two patients with nasopalatine duct cysts at a tertiary care institution with a review of the English medical literature from January 1960 to the present., Methods: A 69-year-old man presented with an asymptomatic swelling of the premaxilla, and a 17-year-old woman presented with a painful swelling of the hard palate. A computed tomography scan, fine-needle aspiration, and preoperative workup were obtained in both cases. Literature was reviewed with respect to epidemiology, etiology, presentation, diagnostic studies, operative management, and recurrence rates., Results: Computed tomography scan demonstrated midline ovoid cystic lesions in both cases. Fine-needle aspiration of both lesions revealed no evidence of malignancy. Surgical treatment consisted of enucleation in the first case and marsupialization in the second case. Both patients did well with no evidence of recurrence. Nasopalatine duct cyst presents in the fourth to sixth decades of life with a male predilection. Recurrence rates range from 0% to 11%., Conclusion: Nasopalatine duct cyst occurs in approximately 1% of the population. Presentation may be asymptomatic or include swelling, pain, and drainage from the hard palate. A well-circumscribed, round, ovoid, or heart-shaped radiolucency is seen on computed tomography. Pathological findings reveal squamous or respiratory cell types, or a combination of these, infiltrated by inflammatory cells. Enucleation is the preferred treatment with low recurrence rates.

Cell separation is a critical process that takes place throughout the life cycle of a plant. It enables roots to emerge from germinating seeds, cotyledons, and leaves to expand, anthers to dehisce, fruit to ripen, and organs to be shed. The focus of this review is to examine how processes such as abscission and dehiscence are regulated and the ways new research strategies are helping us to understand the mechanisms involved in bringing about a reduction in cell-to-cell adhesion. The opportunities for using this information to manipulate cell separation for the benefit of agriculture and horticulture are evaluated.

Histamine affects the balance of T helper type 1 (Th1) and T helper type 2 (Th2) cytokines by shifting cytokine production from a Th1 to a Th2 pattern. Interleukin-13 (IL-13) is an important autacoid mediator that has been implicated in the development of allergic disease. This study was designed to investigate the mechanisms of regulation of IL-13 by histamine in Th2 cells. D10.G4.1 cells, a murine Th2 cell line, were treated with histamine (10(-8)-10(-4) M) and then activated with PMA (phorbol 12 myristate 13-acetate) plus ionomycin or alphaCD3. Levels of IL-13 production were then measured by enzyme-linked immunosorbent assay (ELISA) and semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Cells were pretreated with histamine receptor antagonists pyrilamine, ranitidine, cimetidine and thioperamide to determine the involvement of histamine receptors. Cells were also pretreated with protein kinase A (PKA) inhibitors N-[2-(methylaminoethyl)]-5-isoquinoline-sulfonamide (H-8) and Rp-diastereomer of adenosine cyclic 3'5'-phosphorothionate (Rp-cAMPS), and Janus kinase-signal transducer and activator of transcription (Jak-STAT) inhibitor tyrphostin AG490 prior to the addition of histamine. H-8 is an inhibitor of the catalytic subunit of PKA while Rp-cAMPS is an inhibitor of the regulatory subunit of PKA. Tyrphostin is an inhibitor of Jak2, Jak3, STATI, STAT3 and STAT5. Finally, cells were pretreated with IL-12, a monokine known to repress STAT6 DNA binding. We found that histamine dose-dependently enhanced IL-13 secretion and mRNA levels in Th2 cells via H1 and H2 receptors. Pretreatment of cells with H-8, Rp-cAMPS and tyrphostin prevented histamine-induced secretion and transcription of IL-13. Likewise, pretreatment of Th2 cells with IL-12 also reversed histamine's effects on IL-13 secretion from stimulatory to inhibitory. These observations suggest a role for PKA and the Jak-STAT pathway in histamine-mediated elevation of IL-13 secretion and transcription.

To identify controlling cis acting promoter regions in the B. napus extA extensin gene, expression in transgenic tobacco of 5' - 159, -433, -664, -789 and -940 bp promoter truncations linked to the uidA (B-glucuronidase) reporter coding sequence were analysed. The - 159 and -433 bp truncations directed non specific expression in all cell types within the plant. An activator region which increased expression levels 10 fold in all cell types was located between - 159 to -433 bp. A repressor region was found between -664 to -789 bp; removal of this region resulted in a 15 fold increase in expression. Histochemical analysis showed that transgenics containing the -664, -789 and -940 bp truncations directed expression of the fusion gene only in the phloem. A negative regulatory region located between -433 to -664 bp repressed expression in non-phloem cell types. In areas of the plant subject to tensile stress, the repression exerted by the negative regulatory region was overcome, allowing expression in all cell types. The quantitative repressor and activator regions which controlled absolute expression levels in all cell types were separate from the negative regulatory region which controlled cell type specific expression in response to tensile stress. A wound responsive region was found to be located between -940 to -3500 bp. Thus, the extA gene is under complex control, being regulated by 4 sets of positively and negatively acting cis regions, which control wound inducibility, activation in response to tensile stress, and quantitative expression levels.

Epidemiologic studies have demonstrated a continuing increase in the prevalence of vulvovaginal candidiasis. Although in the past most of these infections were caused by Candida albicans, an increasing percentage are caused by non-albicans Candida species that are less sensitive to the most frequently used antifungal agents. An accurate diagnosis of these infections and the subsequent choice of the most appropriate therapy can only be made after a thorough evaluation of the patient. Successful treatment of vulvovaginal candidiasis is dependent on compliance with therapy; thus, the treatment regimen chosen should fit the patient's daily lifestyle. Newer single-dose regimens offer the option of completing therapy with a single treatment for most patients with uncomplicated vaginal candidiasis. Use of topical agents avoids the potential systemic adverse effects and drug interactions that have been noted with oral antifungals. Patient education and support can also enhance satisfaction with the treatment plan and promote compliance.

A characteristic symmetrical bilateral polyarthritis syndrome is described, affecting the compartements of the wrist joints in a characteristic sequence. The pisiform-triquetral joint gives first, exposing the inferior radioulnar joint so that pisiform pain and tenderness are followed by painful supination. The proximal radiocarpal joint slackens next, followed by the mid-carpal joints. The lunates subluxate as their posterior attachments stretch. Synovial oedema may produce additional median and ulnar carpal tunnel syndromes. The best radiologic sign of this distraction is anterior subluxation of the lunates. Nocturnal arthralgia becomes severe; grip and the ability to write are lost and stretching now shows as a traumatic arthritis in the clavicular joints. The patient is incapacitated and therfore progression halts. A history of excessive straining and lifting is obtained eg with a wheel-barrow, iron pots or strenuous rowing. All serologic tests for rheumatoid disease are negative. Serum uric acid levels and blood sedimentation rates remain normal. There are clear analogies with old descriptions of the effects of torture by stretching from manacles or gauntlets or by the rack.

In contrast to its absence in normal and hyperplastic bronchial mucosa, gamma glutamyl transferase (GGT) has been demonstrated on the luminal surface of adenocarcinoma cells of bronchogenic cancer in three cases studied. On the other hand, little or no GGT was demonstrable in four epidermoid carcinomas and one oat cell cancer. Metaplastic alveolar cells appearing as cuboidal epithelial cells were uniformly positive for GGT and L-homoarginine-sensitive alkaline phosphatase. Whereas interest in GGT has centered in the past on experimental hepatocarcinogenesis, the current results demonstrate an ectopic expression of GGT in metaplastic alveolar cells and in adenocarcinoma of the lung. These findings merit further exploration to document the extent of the expression of GGT in bronchogenic cancer and to attempt to explain its presence in alveolar cell metaplasia.