Silicon-induced changes in viscoelastic properties of sorghum root cell walls.
Silicon is deposited in the endodermal tissue in sorghum (Sorghum bicolor L. Moench) roots. Its deposition is thought to protect vascular tissues in the stele against invasion by parasites and drying soil via hardening of endodermal cells. We studied the silicon-induced changes in mechanical properties of cell walls to clarify the role of silicon in sorghum root. Sorghum seedlings were grown in nutrient solution with or without silicon. The mechanical properties of cell walls were measured in three separated root zones: basal, apical and subapical. Silicon treatment decreased cell-wall extensibility in the basal zone of isolated stele tissues covered by endodermal inner tangential walls. The silicon-induced hardening of cell walls was also measured with increases in elastic moduli (E) and viscosity coefficients (eta). These results provided new evidence that silicon deposition might protect the stele as a mechanical barrier by hardening the cell walls of stele and endodermal tissues. In contrast to the basal zone, silicon treatment increased cell-wall extensibility in the apical and subapical zones with concomitant decrease in E and eta. Simultaneously, silicon promoted root elongation. When root elongation is promoted by silicon, one of the causal factors maybe the silicon-enhanced extensibility of cell walls in the growing zone.


Structural evolution of an enzyme specificity. The structure of rat carboxypeptidase A2 at 1.9-A resolution.
The structure of rat carboxypeptidase A2 (CPA2), which has a unique specificity for tryptophan-containing COOH-terminal peptides, has been determined in an unliganded state at 1.9-A resolution and refined to a crystallographic R-factor of 18.3%. Comparison of the structure of CPA2 with that of bovine carboxypeptidase A (referred to here as CPA1) reveals that the specificity of the former for larger amino acids probably arises from two amino acid replacements within the binding cavity (Thr268----Ala and Leu203----Met), coupled with differences in the positions of conserved residues in a surface loop on one face of the specificity pocket. The position of the reactive-site surface loop may be affected also by a disulfide bridge between Cys210 and Cys244. In this unliganded form of the enzyme, Tyr248 takes up a position interior to the specificity pocket and is distinct from that observed in bovine CPA1. The structural differences between CPA1 and CPA2 correlate strongly with crystallographically determined temperature factors and thus appear to be largest where the enzyme is flexible.


Gallium nitride nanorod arrays as low-refractive-index transparent media in the entire visible spectral region.
Vertically aligned gallium nitride (GaN) nanorod arrays grown by the catalyst-free, self-organized method based on plasma-assisted molecular-beam epitaxy are shown to behave as subwavelength optical media with low effective refractive indices. In the reflection spectra measured in the entire visible spectral region, strong reflectivity modulations are observed for all nanorod arrays, which are attributed to the effects of Fabry-Pérot microcavities formed within the nanorod arrays by the optically flat air/nanorods and nanorods/substrate interfaces. By analyzing the reflectivity interference fringes, we can quantitatively determine the refractive indices of GaN nanorod arrays as functions of light wavelength. We also propose a model for understanding the optical properties of GaN nanorod arrays in the transparent region. Using this model, good numerical fitting can be achieved for the reflectivity spectra.


Fluorescent nanodiamonds enable quantitative tracking of human mesenchymal stem cells in miniature pigs.
Cell therapy is a promising strategy for the treatment of human diseases. While the first use of cells for therapeutic purposes can be traced to the 19th century, there has been a lack of general and reliable methods to study the biodistribution and associated pharmacokinetics of transplanted cells in various animal models for preclinical evaluation. Here, we present a new platform using albumin-conjugated fluorescent nanodiamonds (FNDs) as biocompatible and photostable labels for quantitative tracking of human placenta choriodecidual membrane-derived mesenchymal stem cells (pcMSCs) in miniature pigs by magnetic modulation. With this background-free detection technique and time-gated fluorescence imaging, we have been able to precisely determine the numbers as well as positions of the transplanted FND-labeled pcMSCs in organs and tissues of the miniature pigs after intravenous administration. The method is applicable to single-cell imaging and quantitative tracking of human stem/progenitor cells in rodents and other animal models as well.


Breast cancer education for schoolgirls: an exploratory study.
Adolescent girls are an important target group for breast cancer education and promoting breast awareness. However, research has not established schoolgirls' perceived importance of breast cancer education or explored factors that may impact engagement. This study aimed to identify schoolgirls' concerns about breast cancer, desire to know more and perceived importance of breast cancer education, and explored associations with demographic factors. Of 2089 schoolgirls (11-18 years) surveyed, 1958 completed all relevant breast cancer questions and demographic factors (ethnicity, school type, breast size, physical activity level and age). χ-Tests assessed associations between demographics, desire to know more and perceived importance of breast cancer. Overall, 44% of schoolgirls reported concerns about breast cancer, 72% wanted to know more and 77% rated the topic as extremely important. Breast size was not associated with wanting to know more about breast cancer. Schoolgirls who wanted to know more about breast cancer were White, from single-sex schools with boys at sixth form, more physically active and older. However, among other ethnic groups, school types and physical activity levels, the proportion of girls who wanted to know more about breast cancer was still high (≥61%). This study provides evidence of the need for breast cancer education for schoolgirls across all school types, irrespective of breast size or physical activity levels. The results highlight the need to be inclusive and engage schoolgirls from all ethnic groups and to promote breast awareness at a young age to ensure effective breast cancer education.


Identification of cardiovascular lineage descendants at single-cell resolution.
The transcriptional profiles of cardiac cells derived from murine embryos and from mouse embryonic stem cells (mESCs) have primarily been studied within a cell population. However, the characterization of gene expression in these cells at a single-cell level might demonstrate unique variations that cannot be appreciated within a cell pool. In this study, we aimed to establish a single-cell quantitative PCR platform and perform side-by-side comparison between cardiac progenitor cells (CPCs) and cardiomyocytes (CMs) derived from mESCs and mouse embryos. We first generated a reference map for cardiovascular single cells through quantifying lineage-defining genes for CPCs, CMs, smooth muscle cells (SMCs), endothelial cells (EDCs), fibroblasts and mESCs. This panel was then applied against single embryonic day 10.5 heart cells to demonstrate its ability to identify each endocardial cell and chamber-specific CM. In addition, we compared the gene expression profile of embryo- and mESC-derived CPCs and CMs at different developmental stages and showed that mESC-derived CMs are phenotypically similar to embryo-derived CMs up to the neonatal stage. Furthermore, we showed that single-cell expression assays coupled with time-lapse microscopy can resolve the identity and the lineage relationships between progenies of single cultured CPCs. With this approach, we found that mESC-derived Nkx2-5(+) CPCs preferentially become SMCs or CMs, whereas single embryo-derived Nkx2-5(+) CPCs represent two phenotypically distinct subpopulations that can become either EDCs or CMs. These results demonstrate that multiplex gene expression analysis in single cells is a powerful tool for examining the unique behaviors of individual embryo- or mESC-derived cardiac cells.


A patient-centered medical home model for comprehensive sickle cell care in infants and young children.
The patient-centered medical home (PCMH) has been proposed as a model for comprehensive care coordination and delivery for children with sickle cell disease (SCD), yet little is known regarding the implementation of PCMH core concepts on adherence to preventative care measures, health care utilization, and parent satisfaction. We implemented the newborn cohort clinic (NCC) to explore the application of the PCMH model for infants and children with SCD from birth to age 3 years in 2011. In July 2017, we conducted a retrospective chart review to evaluate subjects currently or previously followed in the clinic. We surveyed parents in the NCC to assess their satisfaction with their experience. A total of 112 patients have been managed in the NCC. All patients received penicillin prophylaxis, while 70% and 73% of patients, respectively, received the 23-valent pneumococcal vaccine and an initial transcranial Doppler by age 36 months. Most (92 of 112) of the subjects utilized the emergency department (569 encounters), with 86% of encounters for fever or other sickle cell-related complications. The majority of parents indicated satisfaction with the clinic, with 71% saying clinic providers always or usually spent enough time with their child, listened carefully to them (81%) and were sensitive to family values and customs (77%). A comprehensive sickle cell clinic as a component of a PCMH is feasible and can achieve high levels of preventative care. Parents are largely satisfied with this model of care.


Global regulation of alternative splicing by adenosine deaminase acting on RNA (ADAR).
Alternative mRNA splicing is a major mechanism for gene regulation and transcriptome diversity. Despite the extent of the phenomenon, the regulation and specificity of the splicing machinery are only partially understood. Adenosine-to-inosine (A-to-I) RNA editing of pre-mRNA by ADAR enzymes has been linked to splicing regulation in several cases. Here we used bioinformatics approaches, RNA-seq and exon-specific microarray of ADAR knockdown cells to globally examine how ADAR and its A-to-I RNA editing activity influence alternative mRNA splicing. Although A-to-I RNA editing only rarely targets canonical splicing acceptor, donor, and branch sites, it was found to affect splicing regulatory elements (SREs) within exons. Cassette exons were found to be significantly enriched with A-to-I RNA editing sites compared with constitutive exons. RNA-seq and exon-specific microarray revealed that ADAR knockdown in hepatocarcinoma and myelogenous leukemia cell lines leads to global changes in gene expression, with hundreds of genes changing their splicing patterns in both cell lines. This global change in splicing pattern cannot be explained by putative editing sites alone. Genes showing significant changes in their splicing pattern are frequently involved in RNA processing and splicing activity. Analysis of recently published RNA-seq data from glioblastoma cell lines showed similar results. Our global analysis reveals that ADAR plays a major role in splicing regulation. Although direct editing of the splicing motifs does occur, we suggest it is not likely to be the primary mechanism for ADAR-mediated regulation of alternative splicing. Rather, this regulation is achieved by modulating trans-acting factors involved in the splicing machinery.


Young children's adaptations to repeated separations from their mothers.
80 infants, toddlers, and preschoolers were observed before, during, and after separations from their mothers, who were attending conferences (M duration = 4 days). Half the sample was separated only once and the other half experienced 3 separations across a 6-month period. The study was designed: (a) to determine how separations affect children's behavior when there are no changes in the family constellation as there typically are during other separations, such as the birth of a new child, and (b) to determine the effects of repeated separations. Fewer changes in sleep and play behaviors suggested that this type of separation was less stressful than separations for the birth of another child. Nonetheless, the separations were still stressful, but principally for the single-separation group. In that group, changes were noted in both play and sleep behaviors. Following reunion, their sleep behaviors and more sophisticated play behaviors returned to baseline. However, activity level and the more insecure behaviors, such as wandering aimlessly, watching other children play, and interacting with their teachers, remained elevated following the mother's return. The multiple-separation group, in contrast, showed only 1 behavior change during their third separation, i.e., reduced interactions with their peers during the separation period, which returned to baseline following reunion. Repeated-measures analyses of the first and third separations of the multiple-separations group suggested that only the first separation was stressful. Thus, the infants and children in this study seemed to adapt to repeated separations.


[Effect of the transcranial electrical stimulation of the endorphinergic brain structures on the functional activity of hepatocytes after toxic exposure].
Biochemical events of the rat hepatocytes cytolysis and deterioration of their synthetic activity and detoxification ability, as well as morphological events of lipid degeneration after acute poisoning with dichloretane and CCl4, were significantly reduced by effects of transcranial stimulation (TES). Blockade of the TES effects with naloxone revealed its endorphinergic nature. Combined effects of the TES and Essenciale preparation were lower than separate those of these agents. The TES effects were clinically corroborated in treatment of toxic hepatitis.


Quantification of the binding affinity of a specific hydroxyapatite binding peptide.
The genesis of bone and teeth involves highly coordinated processes, which involve multiple cell types and proteins that direct the nucleation and crystallization of inorganic hydroxyapatite (HA). Recent studies have shown that peptides mediate the nucleation process, control HA microstructure or even inhibit HA mineralization. Using phage display technology, a short peptide was identified that binds to crystalline HA and to HA-containing domains of human teeth with chemical and morphological specificity. However, the binding affinity and specific amino acids that significantly contribute to this interaction require further investigation. In this study, we employ a microfluidic chip based surface plasmon resonance imaging (SPRi) technique to quantitatively measure peptide affinity by fabricating a novel 4 layer HA SPR sensor. We find the peptide (SVSVGMKPSPRPGGGK) binds with relatively high affinity (K(D) = 14.1 microM +/- 3.8 microM) to HA. The independently measured amino acid fragment SVSV seems to impart a significant contribution to this interaction while the MKPSP fragment may provide a conformational dependent component that enhances the peptides affinity but by itself shows little specificity in the current context. These data show that together, the two moieties promote a stronger synergistic binding interaction to HA than the simple combination of the individual components.


Redox potentials and pKa for benzoquinone from density functional theory based molecular dynamics.
The density functional theory based molecular dynamics (DFTMD) method for the computation of redox free energies presented in previous publications and the more recent modification for computation of acidity constants are reviewed. The method uses a half reaction scheme based on reversible insertion/removal of electrons and protons. The proton insertion is assisted by restraining potentials acting as chaperones. The procedure for relating the calculated deprotonation free energies to Brønsted acidities (pK(a)) and the oxidation free energies to electrode potentials with respect to the normal hydrogen electrode is discussed in some detail. The method is validated in an application to the reduction of aqueous 1,4-benzoquinone. The conversion of hydroquinone to quinone can take place via a number of alternative pathways consisting of combinations of acid dissociations, oxidations, or dehydrogenations. The free energy changes of all elementary steps (ten in total) are computed. The accuracy of the calculations is assessed by comparing the energies of different pathways for the same reaction (Hess's law) and by comparison to experiment. This two-sided test enables us to separate the errors related with the restrictions on length and time scales accessible to DFTMD from the errors introduced by the DFT approximation. It is found that the DFT approximation is the main source of error for oxidation free energies.


Parasite-mediated selection in experimental Daphnia magna populations.
It has been suggested that parasites are a strong selecting force for their hosts and therefore may alter the outcome of competition among host genotypes. We tested the extent to which parasite-mediated selection by different parasite species influenced competition among clones of the cyclic parthenogen Daphnia magna. We monitored clone frequency changes in laboratory microcosm populations consisting of 21 D. magna clones. Parasite treatments (two microsporidians, Glugoides intestinalis and Ordospora colligata) and a parasite-free control treatment were followed over a nine-month period. A further treatment with the bacterium Pasteuria ramosa failed. We found significant differences in clonal success among the treatments: the two parasite treatments differed from the control treatment and from each other. Additionally, we measured the clone-specific population carrying capacity, competitive ability against tester clones, and reproductive success of infected and uninfected females to test whether they correlate with clonal success in the microcosms. The clone-specific competitive ability was a good predictor of clonal success in the microcosms, but clonal carrying capacity and host reproductive success were not. Our study shows that parasite-mediated selection can strongly alter the outcome of clonal competition. The results suggest that parasites may influence microevolution in Daphnia populations during periods of asexual reproduction.


Progress in neuro-imaging of brain tumors.
Magnetic resonance imaging (MRI) is routinely employed in the diagnosis and clinical management of brain tumors. This review provides an overview of the advancements in the field of MRI, with a particular focus on the quantitative assessment by advanced physiological magnetic resonance techniques in light of the new molecular classification of brain tumor. Understanding how molecular phenotypes of brain tumors are reflected in noninvasive imaging is the goal of radiogenomics, which aims at determining the association between imaging features and molecular markers in neuro-oncology. Advanced MRI techniques such as diffusion magnetic resonance imaging and perfusion-weighted imaging add important structural, hemodynamic, and physiological information for tumor diagnosis and classification, as well as to stratify tumor response. Magnetic resonance spectroscopy is able to depict with unprecedented accuracy metabolic biomarkers, which are relevant for molecular subtyping. Ultra-high-field imaging enhances anatomical detail and enables to explore new horizon in tumor imaging. The noninvasive MRI-based assessment of tumor malignancy and molecular status may offer the opportunity to predict prognosis and to select patients who may be candidates for individualized targeted therapies, providing more sensitive tools for their follow-up.


Sound-evoked deflections of outer hair cell stereocilia arise from tectorial membrane anisotropy.
The exceptional performance of mammalian hearing is due to the cochlea's amplification of sound-induced mechanical stimuli. During acoustic stimulation, the vertical motion of the outer hair cells relative to the tectorial membrane (TM) is converted into the lateral motion of their stereocilia. The actual mode of this conversion, which represents a fundamental step in hearing, remains enigmatic, as it is unclear why the stereocilia are deflected when pressed against the TM, rather than penetrating it. In this study we show that deflection of the stereocilia is a direct outcome of the anisotropic material properties of the TM. Using force spectroscopy, we find that the vertical stiffness of the TM is significantly larger than its lateral stiffness. As a result, the TM is more resistant to the vertical motion of stereocilia than to their lateral motion, and so they are deflected laterally when pushed against the TM. Our findings are confirmed by finite element simulations of the mechanical interaction between the TM and stereocilia, which show that the vertical outer hair cells motion is converted into lateral stereocilia motion when the experimentally determined stiffness values are incorporated into the model. Our results thus show that the material properties of the TM play a central and previously unknown role in mammalian hearing.


Molecular Networking-Based Metabolome and Bioactivity Analyses of Marine-Adapted Fungi Co-cultivated With Phytopathogens.
Fungi represent a rich source of bioactive metabolites and some are marketed as alternatives to synthetic agrochemicals against plant pathogens. However, the culturability of fungal strains in artificial laboratory conditions is still limited and the standard mono-cultures do not reflect their full spectrum chemical diversity. Phytopathogenic fungi and bacteria have successfully been used in the activation of cryptic biosynthetic pathways to promote the production of new secondary metabolites in co-culture experiments. The aim of this study was to map the fungal diversity of Windebyer Noor, a brackish lake connected to Baltic Sea (Germany), to induce the chemical space of the isolated marine-adapted fungi by co-culturing with phytopathogens, and to assess their inhibitory potential against six commercially important phytopathogens. Out of 123 marine-adapted fungal isolates obtained, 21 were selected based on their phylogenetic and metabolite diversity. They were challenged with two phytopathogenic bacteria (Pseudomonas syringae and Ralstonia solanacearum) and two phytopathogenic fungi (Magnaporthe oryzae and Botrytis cinerea) on solid agar. An in-depth untargeted metabolomics approach incorporating UPLC-QToF-HRMS/MS-based molecular networking (MN), in silico MS/MS databases, and manual dereplication was employed for comparative analysis of the extracts belonging to nine most bioactive co-cultures and their respective mono-cultures. The phytopathogens triggered interspecies chemical communications with marine-adapted fungi, leading to the production of new compounds and enhanced expression of known metabolites in co-cultures. MN successfully generated a detailed map of the chemical inventory of both mono- and co-cultures. We annotated overall 18 molecular clusters (belonging to terpenes, alkaloids, peptides, and polyketides), 9 of which were exclusively produced in co-cultures. Several clusters contained compounds, which could not be annotated to any known compounds, suggesting that they are putatively new metabolites. Direct antagonistic effects of the marine-adapted fungi on the phytopathogens were observed and anti-phytopathogenic activity was demonstrated.The untargeted metabolomics approach combined with bioactivity testing allowed prioritization of two co-cultures for purification and characterization of marine fungal metabolites with crop-protective activity. To our knowledge, this is the first study employing plant pathogens to challenge marine-adapted fungi.


DNA-PKcs and ATM co-regulate DNA double-strand break repair.
DNA double-strand breaks (DSBs) are repaired by nonhomologous end-joining (NHEJ) and homologous recombination (HR). The NHEJ/HR decision is under complex regulation and involves DNA-dependent protein kinase (DNA-PKcs). HR is elevated in DNA-PKcs null cells, but suppressed by DNA-PKcs kinase inhibitors, suggesting that kinase-inactive DNA-PKcs (DNA-PKcs-KR) would suppress HR. Here we use a direct repeat assay to monitor HR repair of DSBs induced by I-SceI nuclease. Surprisingly, DSB-induced HR in DNA-PKcs-KR cells was 2- to 3-fold above the elevated HR level of DNA-PKcs null cells, and approximately 4- to 7-fold above cells expressing wild-type DNA-PKcs. The hyperrecombination in DNA-PKcs-KR cells compared to DNA-PKcs null cells was also apparent as increased resistance to DNA crosslinks induced by mitomycin C. ATM phosphorylates many HR proteins, and ATM is expressed at a low level in cells lacking DNA-PKcs, but restored to wild-type level in cells expressing DNA-PKcs-KR. Several clusters of phosphorylation sites in DNA-PKcs, including the T2609 cluster, which is phosphorylated by DNA-PKcs and ATM, regulate access of repair factors to broken ends. Our results indicate that ATM-dependent phosphorylation of DNA-PKcs-KR contributes to the hyperrecombination phenotype. Interestingly, DNA-PKcs null cells showed more persistent ionizing radiation-induced RAD51 foci (but lower HR levels) compared to DNA-PKcs-KR cells, consistent with HR completion requiring RAD51 turnover. ATM may promote RAD51 turnover, suggesting a second (not mutually exclusive) mechanism by which restored ATM contributes to hyperrecombination in DNA-PKcs-KR cells. We propose a model in which DNA-PKcs and ATM coordinately regulate DSB repair by NHEJ and HR.


The two-pore domain potassium channel, TWIK-1, has a role in the regulation of heart rate and atrial size.
The two-pore domain potassium (K(+)) channel TWIK-1 (or K2P1.1) contributes to background K(+) conductance in diverse cell types. TWIK-1, encoded by the KCNK1 gene, is present in the human heart with robust expression in the atria, however its physiological significance is unknown. To evaluate the cardiac effects of TWIK-1 deficiency, we studied zebrafish embryos after knockdown of the two KCNK1 orthologues, kcnk1a and kcnk1b. Knockdown of kcnk1a or kcnk1b individually caused bradycardia and atrial dilation (p<0.001 vs. controls), while ventricular stroke volume was preserved. Combined knockdown of both kcnk1a and kcnk1b resulted in a more severe phenotype, which was partially reversed by co-injection of wild-type human KCNK1 mRNA, but not by a dominant negative variant of human KCNK1 mRNA. To determine whether genetic variants in KCNK1 might cause atrial fibrillation (AF), we sequenced protein-coding regions in two independent cohorts of patients (373 subjects) and identified three non-synonymous variants, p.R171H, p.I198M and p.G236S, that were all located in highly conserved amino acid residues. In transfected mammalian cells, zebrafish and wild-type human TWIK-1 channels had a similar cellular distribution with predominant localization in the endosomal compartment. Two-electrode voltage-clamp experiments using Xenopus oocytes showed that both zebrafish and wild-type human TWIK-1 channels produced K(+) currents that are sensitive to external K(+) concentration as well as acidic pH. There were no effects of the three KCNK1 variants on cellular localization, current amplitude or reversal potential at pH7.4 or pH6. Our data indicate that TWIK-1 has a highly conserved role in cardiac function and is required for normal heart rate and atrial morphology. Despite the functional importance of TWIK-1 in the atrium, genetic variation in KCNK1 is not a common primary cause of human AF.


Faculty of Radiation Oncology survey of work practices.
The aim of the present paper was to determine the current working conditions of practising radiation oncologists (RO) in Australasia and their attitudes towards their work and work environment. The authors were requested by the Faculty of Radiation Oncology to conduct a survey of Fellows' work conditions and attitudes. The need for such a survey arose from a workshop of the Faculty held in Sydney in 1995, to determine future directions of the Faculty. Issues of potential interest were identified at the workshop and supplemented by the authors into a survey consisting of both directed and open questions. Respondents were free to remain anonymous. An address list of RO was supplied by the Royal Australian and New Zealand College of Radiologists (RANZCR). Two mailouts were performed to increase the response rate. The survey was completed during the second half of 1996 and analysed in 1997. The response rate was 79% (63-100% according to state/country). The age range was 30-69 years (median: 43 years; mean: 44 years), and 78% of the respondents were male. A public centre was identified as the sole or main place of work for 84% of respondents. The number of RO per practice varied from one to 25 (median: 5). The estimated hours worked per week ranged from 20 to 79 (mean: 52 h; median: 50 h). There were significant differences in allocation of hours between public and private (more clinical hours for private (P = 0.008), more teaching hours for public (P = 0.007)) but no difference in total hours. The responses for clinical work profile were: 'general' 39%, 'largely subspecialty' 37% and both 2% (23% did not respond). The proportion whose practice was 'largely subspecialty' differed between public and private (53% vs 13%, respectively; P = 0.06), and varied according to the number of RO in the practice (62% for > five RO vs 35% for < or = five RO, P = 0.03). The need for subspecialization for the treatment of common tumours (breast, gynaecological etc.) was held by 78% of respondents and was related to the number of RO in the practice (100% for > five RO vs 81% for < or = five RO, P = 0.002). The number of new patients seen per RO in 1995 ranged from 0 to 700 (mean: 342; median: 350), with a significant difference between the mean numbers seen in public versus private settings (331 vs 409, respectively; P = 0.008). Administration of cytotoxics was supervised by 36% of RO across Australasia but was dependent strongly on the state/country of practice (P = 0.0002). The current and preferred roles in clinical management were scored as means of 7.9 and 8.7, respectively, on a linear scale from 1 (mainly technical role) to 10 (significant role in overall management). A total of 40% of RO were content with their current time allocation; the remainder generally wished to reduce clinical time and increase self-education and/or clinical research. Most indicated that they wanted some 'protected time'. Most were content to be called a 'radiation oncologist' and wished to retain this title; the most common alternative was 'clinical oncologist'. Much of the general comment related to perceived loss of control of clinical and academic aspects of oncology to other specialties. The present survey is the first to define current work practices and to explore attitudes to work. It is clear that many RO consider clinical workloads to be excessive, to the exclusion of other work-related activities. There was a strong feeling that significant changes will be required if RO are to maintain the role in cancer management for which they are trained.


Community opinions in the management of corneal ulcers and ophthalmic antibiotics: a survey of 4 states.
To evaluate the ophthalmic community's current opinions of the management of bacterial keratitis and usage of the currently available ophthalmic antibiotics. An anonymous questionnaire was mailed to ophthalmologists in California, Florida, Illinois, and Missouri. The results were tabulated and analyzed statistically. Six hundred twenty-nine usable questionnaires (10.3%) were returned. In the management of corneal ulcers, 42.2% of comprehensive ophthalmologists and 75.3% of cornea specialists perform Gram stains some of the time. A total of 73.3% of comprehensive ophthalmologists and 93.7% of cornea specialists perform corneal cultures some of the time. A total of 88.8% of comprehensive ophthalmologists and 76% of cornea specialists initiate treatment with the newer fluoroquinolone antibiotics. A total of 12.1% of comprehensive ophthalmologists and 41% of cornea specialists would select fortified antibiotics for the treatment of corneal ulcers. The percentage of those who feel that fortified antibiotics are superior ranges from 17.7% for comprehensive ophthalmologists to 33.3% for cornea specialists. A total of 65.7% of comprehensive ophthalmologists indicate that the newer fluoroquinolones have impacted their practices, and 58.3% indicate that they represent an improvement over older fluoroquinolones. Most responding ophthalmologists initiate empiric therapy with the newer fluoroquinolone antibiotics for corneal ulcers, forgoing Gram staining and culturing. However, respondents are not universally sanguine about the newer fluoroquinolones. The practice patterns and opinions on antibiotics differ almost universally between comprehensive ophthalmologists and cornea specialists. Larger, more detailed surveys and more specific analyses would help to further establish the factors that lead to differing management choices and opinions.


Current pharmacogenomic studies on hERG potassium channels.
Genetic polymorphisms in human ether-a-go-go-related gene (hERG) potassium channels are associated with many complex diseases and sensitivity to channel-related drugs. Genotypes may underlie different sensitivities to the same drug, and different drugs selectively repair the functional deficits caused by individual mutations. In fact, not all drugs that block hERG function have adverse effects as previously thought. This suggests that the severe adverse reactions observed clinically may only occur in subjects with a particular genotype, but to others may be safe. Similarly, a drug that is ineffective in one population may be both safe and effective in another. Therefore, detecting polymorphisms in KCNH2 encoding hERG1 is of great significance in guiding the prevention and treatment of related diseases, re-evaluating drug safety, and individualizing treatment. This article reviews current pharmacogenomic studies on hERG potassium channels to provide a reference for developing individualized treatments and evaluating their safety.


Dietary xylo-oligosaccharide improves intestinal functions in weaned piglets.
This study aimed at investigating the effects of dietary xylo-oligosaccharide (XOS) on intestinal functions (i.e., intestinal morphology, tight junctions, gut microbiota and metabolism) and growth performance in weaned piglets. 19 weaned piglets were randomly divided into two groups (n = 9/10): a control group (basic diet) and a XOS treated group in which piglets were fed 0.01% XOS for 28 days. Growth performance, blood cells and biochemical parameters, serum cytokines, intestinal morphology, tight junctions, gut microbiota, and the metabolic profiles of the gut digesta were analyzed. The results showed that dietary supplementation with XOS had little effects on growth performance, blood cells and biochemical parameters, and intestinal morphology. However, the inflammatory status and intestinal barrier were improved in XOS-fed piglets evidenced by the reduction of IFN-γ and upregulation of ZO-1. Microbiota analysis showed that XOS enhanced α-diversity and affected the relative abundances of Lactobacillus, Streptococcus, and Turicibacter at the genus level. The alterations in the microbiota might be further involved in carbohydrate metabolism, cell motility, cellular processes and signaling, lipid metabolism, and metabolism of other amino acids by functional prediction. A metabolomics study identified three differentiated metabolites, including coenzyme Q6, zizyphine A, and pentadecanal, which might be produced by the microbiota and further affect host metabolism. In conclusion, dietary XOS improved the inflammatory status, gut barrier, and microbiota communities, which might be used as a potential feed additive to prevent gut dysfunction caused by weaning in the pig industry.


Role of human neurobehavioural tests in regulatory activity on chemicals.
Psychological performance tests have been used since the mid-1960s in occupational and environmental health toxicology. The interpretation of significantly different test scores in neurobehavioural studies is not straightforward in the regulation of chemicals. This paper sets out some issues which emerged from discussions at an international workshop, organised by the United Kingdom Health and Safety Executive (HSE), to discuss differences in interpretation of human neurobehavioural test data in regulatory risk assessments. The difficulties encountered by regulators confronted with neurobehavioural studies seem to be twofold; some studies lack scientific rigor; other studies, although scientifically sound, are problematic because it is not clear what interpretation to place on the results. Issues relating to each of these points are discussed. Next, scenarios within which to consider the outcomes of neurobehavioural studies are presented. Finally, conclusions and recommendations for further work are put forward.


Cisplatin sensitizes human hepatocellular carcinoma cells, but not hepatocytes and mesenchymal stem cells, to TRAIL within a therapeutic window partially depending on the upregulation of DR5.
Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a member of the TNF family of cytokines and has been shown to induce cell apoptosis in many types of tumors, but not in normal cells. This tumor-selective property has made TRAIL a promising approach for the development of cancer therapy. However, hepatocellular carcinoma (HCC) cells display a striking resistance to TRAIL. Although some chemotherapeutic agents can overcome this resistance, safety issues remain a concern because the combination of these agents and TRAIL has been reported to induce toxicity in normal hepatocytes. In this study, we examined whether cisplatin could reverse TRAIL resistance in HCC cells with different p53 status and evaluated the toxicity of combination TRAIL and cisplatin to normal hepatocytes and mesenchymal stem cells (MSCs). We observed that cisplatin could efficiently sensitize HCC cells, but not hepatocytes and MSCs to TRAIL-induced apoptosis within a wide therapeutic window. The apoptosis of HCC cells only partially depended on the upregulation of DR5 and the status of p53. In addition, we provide favorable evidence supporting the feasibility of the combination of chemotherapy and MSCs transduced with TRAIL.


Anthropometric measures are simple and accurate paediatric weight-prediction proxies in resource-poor settings with a high HIV prevalence.
Accurate weight measurements are essential for both growth monitoring and drug dose calculations in children. Weight can be accurately measured using calibrated scales in resource-rich settings; however, reliable scales are often not available in resource-poor regions or emergency situations. Current age and/or length/height-based weight-prediction equations tend to overestimate weight because they were developed from Western children's measures. To determine the accuracy of several proxy measures for children's weight among a predominately HIV-positive group of children aged 18 months to 12 years in Botswana. Weight, length/height, ulna and tibia lengths, mid-upper arm circumference (MUAC) and triceps skinfold were measured on 775 children recruited from Gaborone, Botswana, between 6 July and 24 August 2011. Mean (95% CI) age and weight were 7.8 years (7.5 to 8.4) and 21.7 kg (21.2 to 22.2), respectively. The majority of children were HIV-positive (n=625, 81%) and on antiretroviral treatment (n=594, 95%). The sample was randomly divided; a general linear model was used to develop weight-prediction equations for one half of the sample (n=387), which were then used to predict the weight of the other half (n=388). MUAC and length/height, MUAC and tibia length and MUAC and ulna length most accurately predicted weight, with an adjusted R2 of 0.96, 0.95 and 0.93, respectively. Using MUAC and length/height, MUAC and tibia length and MUAC and ulna length equations, ≥92% of predicted weight fell within 15% of actual weight, compared with <55% using current equations. The development of nomograms using these equations is warranted to allow for rapid and accurate weight prediction from these simple anthropometric measures in HIV-endemic, resource-constrained settings.


Lung T cells in hypersensitivity pneumonitis: phenotypic and functional analyses.
Cells recovered from bronchoalveolar lavage (BAL) and tissue sections from transbronchial lung biopsies were studied in 16 patients with symptomatic hypersensitivity pneumonitis (HP) and in six subjects with a similar history of exposure but without features of disease by using a series of monoclonal antibodies (MoAb) detecting different lymphocyte subpopulations, including T and T subsets, B lymphocytes, and natural killer (NK) cells. Their functional activities in cytotoxic and suppressor assays and the microenvironment in the lung by using immunohistological techniques were also evaluated. It has been demonstrated that the majority of cells recovered from BAL of HP patients are represented by T8 lymphocytes, with a relevant imbalance of the T4/T8 ratio (p less than 0.001). HNK-1+ cells were markedly increased (p less than 0.001), whereas the frequency of cells bearing other NK-related markers (NK-15, VEP 13, Ab8.28, T10, M1, and Fc gamma R) were not significantly increased with respect to controls. Immunohistological study confirmed that the majority of cells infiltrating lung parenchyma are T8+ lymphocytes. The number of HNK-1+ cells detected on lung biopsies was very low in all cases, even in patients with the highest values on BAL suspensions. The evidence of cells bearing the proliferation-associated markers (Tac and T9 antigens) seems to support the hypothesis of a local proliferation in the lung. In terms of phenotypic analysis, the results observed in the group of asymptomatic individuals are qualitatively superimposable on those observed in the HP group, but the magnitude of the phenomenon is less prominent and therefore the data are not as statistically significant as that produced by the comparison between HP patients and the same controls. Functional analysis of BAL T cells from both HP patients and asymptomatic individuals showed suppressor activity in vitro, as determined by the ability to influence a pokeweed mitogen (PWM)-driven B cell differentiation assay. BAL cells from HP patients were also able to display a definite cytotoxic function in vitro, whereas BAL lymphocytes from asymptomatic subjects did not. Taken together, these data demonstrated that cells responsible for the alveolitis in patients with HP are characterized by the expansion of T cells with the phenotype and functions of both suppressor and/or cytotoxic lymphocytes. This expansion is likely to be related to a local immunologic response to the antigenic stimulus and may provide new insights into the pathogenetic mechanisms of this disease, its pathological pattern, and its management.


Myelopathy due to human T-cell leukemia virus type-1 from the donor after ABO-incompatible liver transplantation.
We report the case of a 53-year-old-man who developed human T-cell leukemia virus type-1-associated myelopathy (HAM) after ABO-incompatible liver transplantation for alcoholic liver cirrhosis. The living donor was seropositive for human T-cell leukemia virus type-1 (HTLV-1) and the recipient was seronegative for HTLV-1 before transplantation. After transplantation, the recipient developed steroid-resistant acute cellular rejection, which was successfully treated using anti-thymocyte globulin, and he was eventually discharged. He underwent spinal surgery twice after the transplantation for the treatment of cervical spondylosis that had been present for a period of 9 months before the transplantation. The surgery improved his gait impairment temporarily. However, his gait impairment progressed, and magnetic resonance imaging revealed multiple sites of myelopathy. He was diagnosed with HAM 16 months after the transplantation. Pulse steroid therapy (1000mg) was administered over a period of 3 days, and his limb paresis improved. Presently, steroid therapy is being continued, with a plan to eventually taper the dose, and he is being carefully followed up at our institution. Our case suggests that liver transplantation involving an HTLV-1-positive living donor carries the risk of virus transmission and short-term development of HAM after transplantation.


Awakening the sleeping giant: anatomy and plasticity of the brain serotonergic system.
The serotonergic neurons of the mammalian brain comprise one of the most expansive chemical systems known. The cell bodies are largely confined to the midline (raphe) region of the brain stem in two general clusters: a superior group that consists of the dorsal raphe nucleus (B-7 and B-6), median raphe nucleus (B-8 and B-5), caudal linear nucleus (rostral B-8), and supralemniscal nucleus (B-9), and an inferior group that consists of nucleus raphe obscurus (B-2), nucleus raphe pallidus (B-1), nucleus raphe magnus (B-3), ventral lateral medulla (B-1/B-3), and the area postrema. The axons from these cells project throughout the neuroaxis from the spinal cord to the olfactory bulb and from the cerebral cortex to the hypothalamus. The development of this giant system begins very early in gestation and is influenced by a variety of growth regulatory factors, including the astroglial protein S-100 beta. Evidence will be presented that the serotonergic system plays a major role in the maturation of the brain by interacting with the 5-HT1A receptors which are most dense during these early developmental periods. The 5-HT1A receptor is located on both neurons and astrocytes, and in the latter cells may serve to stimulate release of S-100 beta. The developmental role of 5-HT appears to become dormant as the brain matures, and during aging and Alzheimer's disease, 5-HT receptors are significantly depressed. However, specific damage to 5-HT fibers in the adult brain by 5,7-dihydroxytryptamine produces a sharp fall in the levels of 5-HT which seems to reactivate the developmental signals in the brain. Not only are the serotonergic fibers encouraged to sprout and expand their territory, but the stimulation of the astrocytic growth factor by a 5-HT1A agonist is reinstated. The ability to recall developmental processes in the adult brain by interrupting the 5-HT fibers may provide important tools for understanding and treating the aged brain.


Inhibition of influenza virus transcription by 2'-deoxy-2'-fluoroguanosine.
The nucleoside analog 2'-deoxy-2'-fluoroguanosine (2'-fluorodGuo) is phosphorylated by cellular enzymes and reversibly inhibits influenza virus replication in chick embryo cells within the first 4 h of infection. RNA hybridization studies revealed that primary and secondary transcription of influenza virus RNA were blocked at a compound concentration of 10 microM, but no inhibition of cell protein synthesis was seen even at high compound concentrations (200 microM). In vitro, the triphosphate of 2'-fluorodGuo is a competitive inhibitor of influenza virus transcriptase activity from disrupted virus, with a Ki of 1.0 microM. The cellular polymerases DNA polymerase alpha and RNA polymerase II were only weakly inhibited or were insusceptible to 2'-fluorodGTP. In kinetic studies with the influenza virus transcriptase, 2'-fluorodGTP, in the absence of GTP, blocked elongation of the virus RNA chain. Similarly, by using purified ribonucleoprotein complexes it was found that the addition of a single nucleotide of 2'-fluorodGTP to the virus RNA caused chain termination, which resulted in the blockage of further virus transcription. Furthermore, the specificity for influenza virus transcriptase was confirmed when the transcriptase from partially resistant virus was found to be 10-fold less susceptible to 2'-fluorodGTP (Ki = 13.1 microM).


Plasma and serum brain-derived neurotrophic factor (BDNF) levels and their association with neurocognition in at-risk mental state, first episode psychosis and chronic schizophrenia patients.
Objectives: Brain-derived neurotrophic factor (BDNF) is involved in numerous cognitive processes. Since cognitive deficits are a core feature of psychotic disorders, the investigation of BDNF levels in psychosis and their correlation with cognition has received increased attention. However, there are no studies investigating BDNF levels in individuals with an at-risk mental state (ARMS) for psychosis. Hence, the aims of the present study were: (1) assessing peripheral BDNF levels across different (potential) stages of psychosis; (2) investigating their association with cognition.Methods: Plasma and serum BDNF levels and neuropsychological performance were assessed in 16 ARMS, six first-episode psychosis (FEP), and 11 chronic schizophrenia (CS) patients. Neuropsychological assessment covered intelligence, verbal memory, working memory, attention and executive functioning.Results: Both plasma and serum BDNF levels were highest in CS, intermediate in FEP and lowest in ARMS. Multiple regression analysis revealed a significant positive association of plasma BDNF levels with planning ability across all groups.Conclusions: The lower peripheral BDNF levels in ARMS compared to FEP and CS might point towards an important drop of this neurotrophin prior to the onset of frank psychosis. The associations of peripheral BDNF with planning-abilities match previous findings.


Exercise prescription for hospitalized people with chronic obstructive pulmonary disease and comorbidities: a synthesis of systematic reviews.
The prescription of physical activity for hospitalized patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) can be complicated by the presence of comorbidities. The current research aimed to synthesize the relevant literature on the benefits of exercise for people with multimorbidities who experience an AECOPD, and ask: What are the parameters and outcomes of exercise in AECOPD and in conditions that are common comorbidities as reported by systematic reviews (SRs)? An SR was performed using the Cochrane Collaboration protocol. Nine electronic databases were searched up to July 2011. Articles were included if they (1) described participants with AECOPD, chronic obstructive pulmonary disease (COPD), or one of eleven common comorbidities, (2) were an SR, (3) examined aerobic training (AT), resistance training (RT), balance training (BT), or a combination thereof, (4) included at least one outcome of fitness, and (5) compared exercise training versus control/sham. This synthesis examined 58 SRs of exercise training in people with AECOPD, COPD, or eleven chronic conditions commonly associated with COPD. Meta-analyses of endurance (aerobic or exercise capacity, 6-minute walk distance--6MWD) were shown to significantly improve in most conditions (except osteoarthritis, osteoporosis, and depression), whereas strength was shown to improve in five of the 13 conditions searched: COPD, older adults, heart failure, ischemic heart disease, and diabetes. Several studies of different conditions also reported improvements in quality of life, function, and control or prevention outcomes. Meta-analyses also demonstrate that exercise training decreases the risk of mortality in older adults, and those with COPD or ischemic heart disease. The most common types of training were AT and RT. BT and functional training were commonly applied in older adults. The quality of the SRs for most conditions was moderate to excellent (>65%) as evaluated by AMSTAR scores. In summary, this synthesis showed evidence of significant benefits from exercise training in AECOPD, COPD, and conditions that are common comorbidities. A broader approach to exercise and activity prescription in pulmonary rehabilitation may induce therapeutic benefits to ameliorate clinical sequelae associated with AECOPD and comorbidities such as the inclusion of BT and functional training.


Range expansion, habitat use, and choosiness in a butterfly under climate change: Marginality and tolerance of oviposition site selection.
Poleward range shifts under climate change involve the colonization of new sites and hence the foundation of new populations at the expanding edge. We studied oviposition site selection in a butterfly under range expansion (Lycaena dispar), a key process for the establishment of new populations. We described and compared the microhabitats used by the species for egg laying with those available across the study sites both in edge and in core populations. We carried out an ecological niche factor analysis (ENFA) to estimate (1) the variety of microhabitats used by the butterfly for egg laying (tolerance) and (2) the extent to which these selected microhabitats deviated from those available (marginality). Microhabitat availability was similar in edge and core populations. Ambient temperature recorded at the site level above the vegetation was on average lower at core populations. In contrast with what is often assumed, edge populations did not have narrower microhabitat use compared to core populations. Females in edge populations even showed a higher degree of generalism: They laid eggs under a wider range of microhabitats. We suggest that this pattern could be related to an overrepresentation of fast deciding personalities in edge populations. We also showed that the thermal time window for active female behavior was reduced in edge populations, which could significantly decrease the time budget for oviposition and decrease the threshold of acceptance during microhabitat selection for oviposition in recently established populations.


[Synchronous (synphaseous) additive models for mitochondrial ATP synthetase complex and for other energy transducing systems of cells (a hypothesis)].
There exist some energy transducing enzymes containing immobile molecules of substrates which are not exchangeable during protein functioning. According to the proposed models the immobile substrates are localized at the "idle" (or "partial" or "imitational") catalytic sites, which differ from normal ("working") active sites of enzymes. Only some steps of a complete reaction sequence which take place at the "working" sites are carried out at the "idle" sites. On the other hand, cyclic conversion of the immobile substrate at an "idle" catalytic site may include some steps which are absent in the "working" site cycle. The occurrence of identical steps on the "idle" and "working" catalytic sites allows to synchronize their action through conformational interconversions of tightly packed and structurally related "idle" and "working" subunits of the enzyme. The presence of covaletly bound substrates or substrates localized in closed cavities of the "idle" sites allows to synchronize the action of many monomers containing such sites due to the absence of the rate-limiting step of simultaneous saturation of many catalytic sites by substrate molecules from solution, and due to the lack of substrate inhibition on the "idle" sites. The functions of the "idle" sites are miscellaneous e.g. in ion-transporting systems these sites are directly involved in ion translocation. In the actomyosin complex the "idle" sites imitate conformational alterations of "working" sites, thus allowing synchronous functioning of the polymeric structure. Variations in the number of the "idle" sites operating simultaneously with one "working" site allow to regulate some parameters of enzymatic processes, e.g. the stoichiometry (number of transported ions per ATP hydrolysed (or synthesised) or electron-transported, or hv-absorbed ones) for ion transported systems or the ratio (velocity of contraction to developed efforts) for the actomyosin complex.


Clinical significance of post-prophylaxis cytomegalovirus infection in lung transplant recipients.
Cytomegalovirus (CMV) disease has been associated with the development of chronic lung allograft dysfunction (CLAD) after transplantation. However, the relevance of CMV replication occurring after the discontinuation of antiviral prophylaxis on the development of CLAD has not been fully established. Patients who underwent lung transplantation during 2004-2014 were included. All patients received antiviral prophylaxis for 3-6 months, followed by monitoring of CMV replication during the first year post-transplantation (preemptive therapy). Risk factors for the development of CLAD were assessed by Cox models. A linear regression model with an interaction coefficient between time and CMV infection was used to evaluate the influence of CMV infection on the evolution of FEV1 . Overall, 69 patients were included, 30/69 (43%) patients developed at least 1 episode of significant CMV infection, and 8/69 (11.5%) patients developed CMV disease. After a median follow-up of 3.67 years, 25/69 (36%) patients developed CLAD and 14/69 (20%) patients died. In the univariate Cox analysis, significant CMV infection (HR 1.177, P = .698), CMV disease (HR 1.001, P = .998), and duration of CMV replication (HR 1.004, P = .758) were not associated with CLAD. Only bacterial pneumonia tended to be associated with CLAD in the multivariate model (HR 2.579, P = .062). We did not observe a significant interaction between CMV replication and evolution FEV1 (interaction coefficient 0.006, CI 95% [-0.084 to 0.096], P = .890). In this cohort of lung transplant recipients receiving antiviral prophylaxis and monitored by preemptive therapy post-prophylaxis, CMV infection did not have impact on long-term allograft lung function.


Transarterial embolization for renal arterial bleeding.
To evaluate the effectiveness of percutaneous transarterial embolization for the treatment of renal arterial bleeding in patients with renovascular injury. The archives of our angiography suite were retrospectively reviewed, and 15 patients who had undergone renal embolization due to renal artery bleeding were included in the study. The site, number, and type of bleeding lesions, and the result of the embolization procedure were recorded. The technical and medical success, and technical and medical complications of the procedure were noted. On renal digital subtraction angiography (DSA) 18 lesions were detected in 15 patients. In 13 cases bleeding was effectively controlled with embolization in a single session. In one case bleeding was controlled on the second attempt. One case underwent nephrectomy. Nontarget embolization was seen in two patients, one treated with polyvinyl alcohol (PVA), the other with n-butyl cyanoacrylate (NBCA) mixture. Iatrogenic dissection of the segmental branch was seen in one patient. Puncture-site bleeding, postembolization syndrome, perirenal abscess or renal abscess, and arterial hypertension were not detected in any of the patients. Percutaneous transarterial embolization is an effective, minimally invasive, and a tissue preserving treatment method for renovascular injuries. Therefore, endovascular embolization should be the first preferred treatment modality.


Structural Dynamics of E6AP E3 Ligase HECT Domain and Involvement of a Flexible Hinge Loop in the Ubiquitin Chain Synthesis Mechanism.
Ubiquitin (Ub) ligases E3 are important factors in selecting target proteins for ubiquitination and determining the type of polyubiquitin chains on the target proteins. In the HECT (homologous to E6AP C-terminus)-type E3 ligases, the HECT domain is composed of an N-lobe and a C-lobe that are connected by a flexible hinge loop. The large conformational rearrangement of the HECT domain via the flexible hinge loop is essential for the HECT-type E3-mediated Ub transfer from E2 to a target protein. However, detailed insights into the structural dynamics of the HECT domain remain unclear. Here, we provide the first direct demonstration of the structural dynamics of the HECT domain using high-speed atomic force microscopy at the nanoscale. We also found that the flexibility of the hinge loop has a great impact not only on its structural dynamics but also on the formation mechanism of free Ub chains.


Transforming growth factor beta signalling and matrix metalloproteinases in the mucosa overlying Crohn's disease strictures.
In addition to its crucial role in dampening tissue-damaging immune responses in the gut, transforming growth factor beta (TGFbeta) is a potent profibrogenic agent inducing collagen synthesis and regulating the balance between matrix-degrading matrix metalloproteinases (MMPs) and their inhibitors (TIMPs). TGFbeta signalling was investigated by analysis of Smad proteins and MMPs/TIMPs in the mucosa overlying strictures in patients with Crohn's disease (CD). Specimens were collected from macroscopically normal mucosa overlying strictured and non-strictured gut of patients with fibrostenosing CD. Isolated myofibroblasts were cultured with anti-TGFbeta blocking antibody or TGF beta 1. TGFbeta transcripts were analysed by quantitative reverse transcription-PCR (RT-PCR). Smad proteins and MMPs were determined by immunoblotting. MMP-12 activity was measured by a real-time MMP-12 activity assay. An in vitro wound-healing scratch assay was used to assess myofibroblast migration. TGFbeta transcripts, phosphorylated Smad2-Smad3 (pSmad2-3) and TIMP-1 proteins were higher in mucosa overlying strictures than in mucosa overlying non-strictured areas. In contrast, mucosa overlying strictured gut had lower expression of Smad7, MMP-12 and MMP-3. Myofibroblasts from mucosa overlying strictured gut showed higher TGFbeta transcripts, a greater pSmad2-3 response to TGFbeta, increased TIMP-1, lower Smad7, increased collagen production and reduced migration ability compared with myofibroblasts from mucosa overlying non-strictured gut. TGFbeta blockade increased myofibroblast MMP-12 production and migration, more obviously in myofibroblasts isolated from mucosa overlying non-strictured compared with strictured gut. Changes in TGF-beta signalling and MMP production were identified in the mucosa overlying strictures in CD which may give a window into the process of fibrosis.


Late bronchoplasty for missed traumatic rupture of bronchus.
Two patients of traumatic rupture of main bronchus are reported here. One patient presented 2 years after blunt chest trauma while the other patient presented 9 weeks after trauma. Lesion in both patients were missed in the immediate post-trauma period. In both patients pulmonary angiogram demonstrated normal vasculature in the atelectatic lung, while intraoperatively good compliance and elasticity were found. Bronchoplasty done in both patients had good postoperative results.


Blood flow regulation by S-nitrosohemoglobin in the physiological oxygen gradient.
The binding of oxygen to heme irons in hemoglobin promotes the binding of nitric oxide (NO) to cysteinebeta93, forming S-nitrosohemoglobin. Deoxygenation is accompanied by an allosteric transition in S-nitrosohemoglobin [from the R (oxygenated) to the T (deoxygenated) structure] that releases the NO group. S-nitrosohemoglobin contracts blood vessels and decreases cerebral perfusion in the R structure and relaxes vessels to improve blood flow in the T structure. By thus sensing the physiological oxygen gradient in tissues, hemoglobin exploits conformation-associated changes in the position of cysteinebeta93 SNO to bring local blood flow into line with oxygen requirements.


Co-expression and promoter content analyses assign a role in biotic and abiotic stress responses to plant natriuretic peptides.
Plant natriuretic peptides (PNPs) are a class of systemically mobile molecules distantly related to expansins. While several physiological responses to PNPs have been reported, their biological role has remained elusive. Here we use a combination of expression correlation analysis, meta-analysis of gene expression profiles in response to specific stimuli and in selected mutants, and promoter content analysis to infer the biological role of the Arabidopsis thaliana PNP, AtPNP-A. A gene ontology analysis of AtPNP-A and the 25 most expression correlated genes revealed a significant over representation of genes annotated as part of the systemic acquired resistance (SAR) pathway. Transcription of these genes is strongly induced in response to salicylic acid (SA) and its functional synthetic analogue benzothiadiazole S-methylester (BTH), a number of biotic and abiotic stresses including many SA-mediated SAR-inducing conditions, as well as in the constitutive SAR expressing mutants cpr5 and mpk4 which have elevated SA levels. Furthermore, the expression of AtPNP-A was determined to be significantly correlated with the SAR annotated transcription factor, WRKY 70, and the promoters of AtPNP-A and the correlated genes contain an enrichment in the core WRKY binding W-box cis-elements. In constitutively expressing WRKY 70 lines the expression of AtPNP-A and the correlated genes, including the SAR marker genes, PR-2 and PR-5, were determined to be strongly induced. The co-expression analyses, both in wild type and mutants, provides compelling evidence that suggests AtPNP-A may function as a component of plant defence responses and SAR in particular. The presented evidence also suggests that the expression of AtPNP-A is controlled by WRKY transcription factors and WRKY 70 in particular. AtPNP-A shares many characteristics with PR proteins in that its transcription is strongly induced in response to pathogen challenges, it contains an N-terminal signalling peptide and is secreted into the extracellular space and along with PR-1, PR-2 and PR-5 proteins it has been isolated from the Arabidopsis apoplast. Based on these findings we suggest that AtPNP-A could be classified as a newly identified PR protein.


Communication: Water activation and splitting by single metal-atom anions.
We report experimental and computational results pertaining to the activation and splitting of single water molecules by single atomic platinum anions. The anion photoelectron spectra of [Pt(H2O)]-, formed under different conditions, exhibit spectral features that are due to the anion-molecule complex, Pt-(H2O), and to the reaction intermediates, HPtOH- and H2PtO-, in which one and two O-H bonds have been broken, respectively. Additionally, the observations of PtO- and H2 + in mass spectra strongly imply that water splitting via the reaction Pt- + H2O → PtO- + H2 has occurred. Extending these studies to nickel and palladium shows that they too are able to activate single water molecules, as evidenced by the formation of the reaction intermediates, HNiOH- and HPdOH-. Computations at the coupled cluster singles and doubles with perturbatively connected triples level of theory provide structures and vertical detachment energies (VDEs) for both HMOH- and H2MO- intermediates. The calculated and measured VDE values are in good agreement and thus support their identification.


Detection and quantitation of intermittent CSF rhinorrhea during prolonged cisternography with 111In-DTPA.
In an attempt to improve the diagnosis and to assess the intensity of intermittent cerebrospinal fluid rhinorrhea, indium-111 uptake by nasal pledgets during prolonged cisternography using 111In-DTPA was determined. In 12 patients having suspected cerebrospinal fluid rhinorrhea, cisternograms with calculation of counts/pixel in regions of interest were obtained, blood samples drawn, and nasal pledgets placed several times during 48 or 72 hr after lumbar intrathecal administration of 111In-DTPA. In three patients intermittent cerebrospinal fluid rhinorrhea was thus revealed; study by computed tomography was negative in each case. This procedure of extended radionuclide cisternography helped to detect, localize, and quantify intermittent cerebrospinal fluid rhinorrhea so that therapeutic decisions were facilitated and surgical intervention was minimized.


A case of rotenone poisoning from ingesting Derris trifoliata Lour. (Tuba fruit/pod) in Malaysia.
Derris trifoliata is mainly found in mangrove area in tropical regions and the plant extract is traditionally used for fishing by poisoning. This is the first case report of rotenone poisoning in a child from ingesting Derris trifoliata seed. The child developed altered consciousness, vomiting, hypotension, metabolic acidosis, and acute kidney injury. Species identification of this case requires the collaborative efforts of various agencies. She survived from the poisoning with no neurological sequelae.


Receptor-Ligand Rebinding Kinetics in Confinement.
Rebinding kinetics of molecular ligands plays a key role in the operation of biomachinery, from regulatory networks to protein transcription, and is also a key factor in design of drugs and high-precision biosensors. In this study, we investigate initial release and rebinding of ligands to their binding sites grafted on a planar surface, a situation commonly observed in single-molecule experiments and that occurs in vivo, e.g., during exocytosis. Via scaling arguments and molecular dynamic simulations, we analyze the dependence of nonequilibrium rebinding kinetics on two intrinsic length scales: the average separation distance between the binding sites and the total diffusible volume (i.e., height of the experimental reservoir in which diffusion takes place or average distance between receptor-bearing surfaces). We obtain time-dependent scaling laws for on rates and for the cumulative number of rebinding events. For diffusion-limited binding, the (rebinding) on rate decreases with time via multiple power-law regimes before the terminal steady-state (constant on-rate) regime. At intermediate times, when particle density has not yet become uniform throughout the diffusible volume, the cumulative number of rebindings exhibits a novel, to our knowledge, plateau behavior because of the three-dimensional escape process of ligands from binding sites. The duration of the plateau regime depends on the average separation distance between binding sites. After the three-dimensional diffusive escape process, a one-dimensional diffusive regime describes on rates. In the reaction-limited scenario, ligands with higher affinity to their binding sites (e.g., longer residence times) delay entry to the power-law regimes. Our results will be useful for extracting hidden timescales in experiments such as kinetic rate measurements for ligand-receptor interactions in microchannels, as well as for cell signaling via diffusing molecules.


[Nature of postoperative treatment of patients with pancreas operations].
The work is devoted to the employment of a prolonged selective bringing of medical drugs to hepatopancreatoduodenal zone in complex with other methods of prophylactics and treatment of postoperative pancreatites in patients operated on the pancreas. The duration of staying the catheters in the celiac artery was 72-96 h at an average. Thanks to introduction of the method of selective therapy the amount of complications was decreased after planned operations on organs of the pancreatoduodenal zone by 12,6% and after operations for pancreatonecrosis by 19,5%.


Comparative Study on the Efficacy of Precise Radiotherapy in Lymphatic Drainage Area of Esophageal Cancer between Preventive Radiation and Invasive Field Radiation.
To investigate the efficacy of precise radiation therapy for lymphatic drainage area prevention (ENI) and invasive field irradiation (IFI) in patients with esophageal cancer undergoing radical radiotherapy. A retrospective analysis was conducted on 96 esophageal cancer patients admitted to our hospital from March 2018 to March 2021 who underwent radical radiotherapy. Among them, 48 patients who received precise radiation therapy to prevent radiation in the lymphatic drainage area were included in the ENI group, and 48 patients who received field irradiation were included in the IFI group. Compare and analyze the total clinical response rate, local control rate within 3 years after treatment, survival rate, and incidence of adverse reactions after radiotherapy between two groups of patients. There was no statistically significant difference in the total clinical response rate between the ENI group and the IFI group after radiotherapy (P > .05). There was no statistically significant difference in local control rate and survival rate between the ENI group and the IFI group within 3 years of treatment (P > .05). The incidence of postoperative complications in the ENI group was significantly higher than that in the IFI group, with a statistically significant difference (P < .05). When giving radical radiotherapy to esophageal cancer patients, the clinical total effective rate, survival rate, and local control rate of involving field irradiation are equivalent to the preventive radiation effect of precise radiotherapy for lymphatic drainage area. Besides, involving field irradiation can reduce the incidence of adverse reactions in patients after radiotherapy, which has high clinical value.


Geographic variation in dengue seroprevalence and force of infection in the urban paediatric population of Indonesia.
Understanding the heterogeneous nature of dengue transmission is important for prioritizing and guiding the implementation of prevention strategies. However, passive surveillance data in endemic countries are rarely adequately informative. We analyzed data from a cluster-sample, cross-sectional seroprevalence study in 1-18 year-olds to investigate geographic differences in dengue seroprevalence and force of infection in Indonesia. We used catalytic models to estimate the force of infection in each of the 30 randomly selected sub-districts. Based on these estimates, we determined the proportion of sub-districts expected to reach seroprevalence levels of 50%, 70% and 90% by year of age. We used population averaged generalized estimating equation models to investigate individual- and cluster-level determinants of dengue seropositivity. Dengue force of infection varied substantially across Indonesia, ranging from 4.3% to 30.0% between sub-districts. By age nine, 60% of sub-districts are expected to have a seroprevalence ≥70%, rising to 83% by age 11. Higher odds of seropositivity were associated with higher population density (OR = 1.54 per 10-fold rise in population density, 95% CI: 1.03-2.32) and with City (relative to Regency) administrative status (OR = 1.92, 95% CI: 1.32-2.79). Our findings highlight the substantial variation in dengue endemicity within Indonesia and the importance of understanding spatial heterogeneity in dengue transmission intensity for optimal dengue prevention strategies including future implementation of dengue vaccination programmes.


Pulmonary aspiration: new therapeutic approaches in the experimental model.
Acute lung injury caused by gastric aspiration is a frequent occurrence in unconscious patients. Acute respiratory distress syndrome in association with gastric aspiration carries a mortality of up to 30% and accounts for up to 20% of deaths associated with anesthesia. Although the clinical condition is well known, knowledge about the exact inflammatory mechanisms is still incomplete. This study was performed to define the role of alveolar macrophages in this inflammatory response. In addition, potentially modifying effects of intratracheally applied nuclear factor kappaB inhibitor pyrrolidine dithiocarbamate were investigated. Rat alveolar macrophages were depleted by intratracheal administration of clodronate liposomes, and lung injury was evaluated 6 h after instillation of 0.1N hydrochloric acid. In a second set of experiments, pyrrolidine dithiocarbamate was intratracheally instilled 3 h after hydrochloric acid application, and injury parameters were determined. Depletion of alveolar macrophages resulted in decreased production of inflammatory mediators in acid aspiration (23-80% reduction of messenger RNA or protein of inflammatory mediators; P < 0.05) and consequently also in diminished neutrophil recruitment (36% fewer neutrophils; P < 0.01). Treatment with pyrrolidine dithiocarbamate was highly effective in decreasing neutrophil recruitment (66%; P < 0.01) and vascular permeability (80%; P < 0.001). These data suggest that alveolar macrophages play an essential role in the inflammatory response of acid-induced lung injury. For the first time, attenuation of acid-induced lung injury with an inhibitor, applied after the onset of injury, is shown.


Is Europe prepared to meet the oral health needs of older people?
To discuss the preparedness of the social and health care systems and the health workforce in Europe to manage the increasing general and oral health care needs of older adults. There are large inequalities across European countries and regions in the demographic, socioeconomic and health status of the elderly. The ageing of the population and the economic crisis put at risk the existing social and health care systems and are expected to further widen the existing inequalities. Despite the increase in funding for the general health care, public funding for dental care has reduced, limiting the access for the disadvantaged elderly. Dental care is isolated from health care policies and funding. At the same time there is a significant shortage of adequately trained personnel in the care of the elderly and a shortage of training opportunities particularly at a postgraduate and continuing education level. Immediate action is needed and appropriate strategies need to be implemented. Oral health prevention, delivery policies and funding should be integrated within the general health care system. Clinical protocols and guidelines need to be developed on the oral care of the elderly. Interdisciplinary training in the care of the elderly needs to be implemented for all health care workers (dentists, physicians, nurses, health care aids, social workers) at all education levels to enhance comprehensive care.


Seamless support from screening to anti-HCV treatment and HCC/ decompensated cirrhosis: Subsidy programs for HCV elimination.
Viral hepatitis poses a major public health problem in Japan. Chronic viral hepatitis is a progressive liver disease that eventually develops into liver cirrhosis and liver cancer. Since nucleic acid analog therapy for hepatitis B and interferon-free therapy for hepatitis C have made it possible to control the disease status or eliminate the viruses, it is very important that more people receive hepatitis virus tests to confirm the presence of infection at an early stage, and that patients with hepatitis detected by the tests receive appropriate medical care. Currently, the government of Japan is implementing comprehensive measures for hepatitis control based on five key strategies. Moreover, the goal listed in the Basic Guidelines on Hepatitis Measures is to reduce the frequency of progression of hepatitis to cirrhosis or liver cancer through a scheme consisting of testing people for hepatitis, getting those who test positive to visit a medical institution and receive treatment, and providing appropriate and high-quality hepatitis care through specialized medical institutions and regional core centers for the management of liver disease. To achieve the goal, various subsidy programs including an expense subsidy system for hepatitis treatment have been implemented in Japan. It is important for healthcare professionals to have sufficient knowledge of public support for efficient hepatitis C virus (HCV)-related liver disease detection and care.


Preparation and crystallization of the Grb7 SH2 domain in complex with the G7-18NATE nonphosphorylated cyclic inhibitor peptide.
Grb7 is an adapter protein that is involved in signalling pathways that mediate eukaryotic cell proliferation and migration. Its overexpression in several cancer types has implicated it in cancer progression and led to the development of the G7-18NATE cyclic peptide inhibitor. Here, the preparation of crystals of G7-18NATE in complex with its Grb7 SH2 domain target is reported. Crystals of the complex were grown by the hanging-drop vapour-diffusion method using PEG 3350 as the precipitant at room temperature. X-ray diffraction data were collected from crystals to 2.4 Å resolution using synchrotron X-ray radiation at 100 K. The diffraction was consistent with space group P2(1), with unit-cell parameters a=52.7, b=79.1, c=54.7 Å, α=γ=90.0, β=104.4°. The structure of the G7-18NATE peptide in complex with its target will facilitate the rational development of Grb7-targeted cancer therapeutics.


Latex allergy. Prevention and treatment.
Latex allergy is a significant, life-threatening problem for some patients and health care workers. This article identifies those individuals at greatest risk for latex allergy and describes measures that can be implemented in order to provide a latex-free environment. Currently accepted pretreatment protocols are presented and guidelines for the treatment of anaphylaxis, should it occur, are reviewed.


Stability of enterocytes and certain enzymatic activities in suspensions of cells from the villous tip to the crypt of Lieberkühn of the mouse small intestine.
A series of studies in this laboratory have focused on how an indigenous microbiota influences the activities of alkaline phosphatase, phosphodiesterase I, and thymidine kinase in the enterocytes of the upper small intestine of mice. To draw conclusions about the role of the microflora in determining levels of enzymatic activity, we found it necessary to develop a procedure by which cell suspensions could be obtained containing enterocytes isolated sequentially from the villous tip to the crypt of Lieberkühn. The procedure was modified from the one developed for rats by Weiser (J. Biol. Chem. 248:2536-2541, 1973), involved a minimum number of interfering factors (e.g., proteolytic enzymes and mechanical agitation), and worked reproducibly for mice. During development of the procedure, some variables affecting the assays of the enzymes known to be present in enterocytes were also explored. Rods to which were tied everted segments of gut were incubated in a series of tubes containing a solution of EDTA the concentration of which was changed from 1.5 to 5.0 mM, thus giving a greater yield of enterocytes at every step. The cells incubating in the chelating solution were most stable when 0.23 M sucrose was included in the EDTA solutions. Success in assaying enzymatic activities in the cell suspensions depended on (i) how the cells were isolated, (ii) the assay procedure for thymidine kinase, and (iii) whether cellular suspensions or extracts were assayed.


The Efficacy of Dienogest in the Treatment of Simple Endometrial Hyperplasia without Atypia.
The aim of the study was to compare the effectiveness of dienogest (DIE), depo medroxyprogesterone 17-acetate (MPA), and micronized progesteron (MP) regimens for treatment of simple endometrial hyperplasia (EH) without atypia. One hundred and twenty premenopausal patients aged between 35 and 55, with simple EH without atypia, were offered 3 types of progestins (MPA, MP, and DIE) and were randomized to one of them. After 6 months of treatment, patients were reevaluated. The efficacy of different progestins in treatment of EH was compared with regard to resolution and regression rates. We classified the terms of resolution and regression as "complete response" and persistence as "no response". A total of 99 patients continued the study (31 in MP, 35 in MPA, and 33 in the DIE group). None of the results of endometrial pathology were progressed to atypia or complex hyperplasia. The complete response resolution rate was 93.5% in the MP, 88.5% in the MPA, and 96.9% in the DIE group. The highest complete response rate was in the DIE group, although there was no statistically significant difference between groups (p = 0.39). The efficacy of progestins was similar between the groups. DIE is an effective and convenient method for thinning the endometrium.


Bi-allelic loss of function variants in SLC30A5 as cause of perinatal lethal cardiomyopathy.
Perinatal mortality is a heavy burden for both affected parents and physicians. However, the underlying genetic causes have not been sufficiently investigated and most cases remain without diagnosis. This impedes appropriate counseling or therapy. We describe four affected children of two unrelated families with cardiomyopathy, hydrops fetalis, or cystic hygroma that all deceased perinatally. In the four patients, we found the following homozygous loss of function (LoF) variants in SLC30A5 NM_022902.4:c.832_836del p.(Ile278Phefs*33) and NM_022902.4:c.1981_1982del p.(His661Tyrfs*10). Knockout of SLC30A5 has previously been shown a cardiac phenotype in mouse models and no homozygous LoF variants in SLC30A5 are currently described in gnomAD. Taken together, we present SLC30A5 as a new gene for a severe and perinatally lethal form of cardiomyopathy.


Developing Pandemic Comics for Youth Audiences.
In spring 2020 our team received funding from the Rapid Response Research program of the National Science Foundation to develop comics that would help youth understand the COVID-19 pandemic. Our project built on a decade of expertise creating comics about the biology of viruses. In collaboration with virologists and artists, we developed three comic stories about COVID-19 during the pandemic and posted them on-line during the last half of 2020. The fictional narratives address fundamental issues in biology, virology, and network science, in order to help readers understand the complexities of living through a viral pandemic. The stories focus on three themes: the biology and social context of the COVID-19 virus; the relationship of wild animals, particularly bats, to the pandemic; and the impact of the pandemic on Tribal communities. We describe the challenges of comic development during the pandemic and the feedback from youth on whether the comics were appealing, interesting, and understandable. The stories were posted on worldofviruses.unl.edu and are published by the University of Nebraska Press as the book, C'RONA Pandemic Comics, with essays for youth about the virus and the pandemic.


[Diabetes mellitus mortality in the Province of Córdoba: during the 1986-1995 period].
The fact that Diabetes Mellitus has become one of the relevant health problems and is besides an important death cause in the world, has led to investigate mortality as produced by this pathology in the Province of Córdoba during the period 1986/1995. Death register because of Diabetes Mellitus for the whole province(for years 1986-1995) were obtained from the Stadistic Department of the Health Ministry of the Province of Córdoba. Brute and specific rates based on sex and age groups were determined from a total of 6.718 deceased (3,300 males and 3,418 females), evolution of the same and its relation with diverse variables such as instruction level, poor housing and U. B. N.(Unsatisfied Basic Needs). Result showed that death rate increased during the period Studied, 24.42/100,000 being the medium annual rate, with no significative difference between both sexes (P = 0.76 > 0.05). In relation to age, rates raised according to growing age, with prevalence of the masculine, in general from 45 years and onward, except for 1986, 1989 and 1995 years when feminine death rate is greater in the 75 years-old and onward group. With regard to the other variables considered, when comparing them with the social and economic development of each Department in the Province of Córdoba, we could infer that industrialization, urbanization and social and economic evolution of the same have a positive correlation with mortality caused by diabetes.


Interplay between spin-crossover and luminescence in a multifunctional single crystal iron(ii) complex: towards a new generation of molecular sensors.
We present a new example of a mononuclear iron(ii) complex exhibiting a correlated spin-crossover (SCO) transition and strong fluorescence, whose coordination sphere is saturated, for the first time, by six phosphorescent ligands. The interplay between SCO and light emission properties in the thermal region of the spin transition was investigated by means of magnetic, fluorescence, optical absorption and optical microscopy measurements on a single crystal. Overall, the results show an excellent correlation between fluorescence and magnetic data of the present gradual transition, indicating an extreme sensitivity of the optical activity of the ligand to the spin state of the active iron(ii) ions. These results open the way for conceiving new prototypes of pressure and temperature sensors based on this synergy between SCO and luminescence properties. In particular, the fact that cooperative SCO material is not a prerequisite for obtaining such synergetic effects, is useful for the design of thin films or nanoparticles, in which the cooperativity is reduced, for appropriate implementation in nanosized devices to enhance the sensing properties at the nanoscale.


Polygenic risk for Alzheimer's disease is not associated with cognitive ability or cognitive aging in non-demented older people.
Alzheimer's disease (AD) and non-pathological cognitive aging have phenotypic similarities which may be influenced by an overlapping set of genetic variants. Genome-wide complex trait analysis estimates that common genetic variants account for about 24% of the variation contributing to liability for AD. It is also estimated that 24% of the variance of non-pathological cognitive aging is accounted for by common single nucleotide polymorphisms. However, although the APOE locus is associated with both AD and cognitive aging, it is not known to what extent other common genetic variants, with smaller effect sizes that influence both, overlap. We test the hypothesis that polygenic risk for AD is associated with cognitive ability and cognitive change in about 3,000 non-demented older people (Cognitive Ageing Genetics England and Scotland-CAGES-consortium). We found no significant association of polygenic risk for AD with cognitive ability or cognitive change in CAGES, indicating that the genetic etiologies of AD and non-pathological cognitive decline differ.


A mechanistic niche model for measuring species' distributional responses to seasonal temperature gradients.
Niche theory is central to understanding how species respond geographically to climate change. It defines a species' realized niche in a biological community, its fundamental niche as determined by physiology, and its potential niche--the fundamental niche in a given environment or geographic space. However, most predictions of the effects of climate change on species' distributions are limited to correlative models of the realized niche, which assume that species are in distributional equilibrium with respect to the variables or gradients included in the model. Here, I present a mechanistic niche model that measures species' responses to major seasonal temperature gradients that interact with the physiology of the organism. I then use lethal physiological temperatures to parameterize the model for bird species in North and South America and show that most focal bird species are not in direct physiological equilibrium with the gradients. Results also show that most focal bird species possess broad thermal tolerances encompassing novel climates that could become available with climate change. I conclude with discussion of how mechanistic niche models may be used to (i) gain insights into the processes that cause species to respond to climate change and (ii) build more accurate correlative distribution models in birds and other species.


Memory CD4+ T Cells in Immunity and Autoimmune Diseases.
CD4+ T helper (Th) cells play central roles in immunity in health and disease. While much is known about the effector function of Th cells in combating pathogens and promoting autoimmune diseases, the roles and biology of memory CD4+ Th cells are complex and less well understood. In human autoimmune diseases such as multiple sclerosis (MS), there is a critical need to better understand the function and biology of memory T cells. In this review article we summarize current concepts in the field of CD4+ T cell memory, including natural history, developmental pathways, subsets, and functions. Furthermore, we discuss advancements in the field of the newly-described CD4+ tissue-resident memory T cells and of CD4+ memory T cells in autoimmune diseases, two major areas of important unresolved questions in need of answering to advance new vaccine design and development of novel treatments for CD4+ T cell-mediated autoimmune diseases.


Implication of the product of the bovine herpesvirus type 1 UL25 gene in capsid assembly.
The UL25 ORF of bovine herpesvirus type 1 (BHV-1) was demonstrated recently to represent a gene encoding a 63 kDa viral protein. To investigate the role of this gene in virus replication, a BHV-1 UL25 deletion mutant was constructed. Although the UL25 mutant synthesizes late viral proteins and viral DNA, it fails to produce virus progeny in cells that do not express the UL25 gene, demonstrating that the UL25 protein is essential for the replicative cycle of BHV-1. Moreover, Southern blotting analyses of HindIII-digested DNA from infected non-complementing cells probed with the leftward terminal fragment of the BHV-1 linear genome revealed that the cleavage of the viral DNA produced is not impaired. However, the packaging of this cleaved DNA is compromised severely, since only few full C capsids were observed in infected non-complementing cells by transmission electron microscopy.


Detection of a second t(14;18) breakpoint cluster region in human follicular lymphomas.
Our results indicate that there are two major breakpoint cluster regions in chromosome 18 DNA for t(14;18) translocations in follicular lymphomas. The absence of a pFL-1 homologous transcript in a cell line containing a pFL-2-detectable translocation suggests that there may be two different pathogenetic consequences of t(14;18) translocations. One possibility is that, despite the distances between them (greater than 20 kb), breakpoints in the two cluster regions in some way affect transcription of the same gene product, which has not yet been identified. Alternatively, two separate transcriptional units may be involved. The availability of DNA probes for each of the two t(14;18) breakpoint cluster regions will allow further studies regarding the biologic significance of these two genetically distinct classes of t(14;18) translocations.


Stereocontrolled synthesis of psi[CH = CH] dipeptide isosteres.
A new stereocontrolled synthesis of the psi[CH = CH] dipeptide isostere is described. They key step of the sequence relies on the stereospecific alpha-alkylation of delta-amino-gamma-mesyloxy-alpha,beta-unsaturated esters. The broad availability of nucleophilic alpha-side chains by this method allows the preparation of a wide variety of psi[CH = CH] isosteres with predictable stereochemistry.


White matter aberrations in prepubertal estrogen-naive girls with monosomic Turner syndrome.
Turner syndrome (TS) offers a unique opportunity to investigate associations among genes, the brain, and cognitive phenotypes. In this study, we used 3 complementary analyses of diffusion tensor imaging (DTI) data (whole brain, region of interest, and fiber tractography) and a whole brain volumetric imaging technique to investigate white matter (WM) structure in prepubertal, nonmosaic, estrogen-naive girls with TS compared with age and sex matched typically developing controls. The TS group demonstrated significant WM aberrations in brain regions implicated in visuospatial abilities, face processing, and sensorimotor and social abilities compared with controls. Extensive spatial overlap between regions of aberrant WM structure (from DTI) and regions of aberrant WM volume were observed in TS. Our findings indicate that complete absence of an X chromosome in young females (prior to receiving exogenous estrogen) is associated with WM aberrations in specific regions implicated in characteristic cognitive features of TS.


Oxygen and carbon dioxide regulation of toxic shock syndrome toxin 1 production by Staphylococcus aureus MN8.
The production of toxic shock syndrome toxin 1 (TSST-1) by Staphylococcus aureus MN8 exposed to a range of oxygen concentrations (0 to 21% [vol/vol]) was examined in batch and thin-film cultures. The response of S. aureus to this range of oxygen concentrations was studied in the absence and in the presence of 7% (vol/vol) carbon dioxide. In the absence of carbon dioxide, TSST-1 production in batch cultures increased from negligible levels in the presence of oxygen concentrations of 1% or less to 500 ng/ml in the presence of 2% oxygen and then decreased to 70 ng/ml or less in the presence of oxygen concentrations of 6% and higher. In the presence of carbon dioxide, however, toxin production increased from negligible levels in the presence of 1% oxygen to 1,900 ng/ml in the presence of 21% oxygen. In thin-film cultures, TSST-1 production increased from nearly undetectable levels under anaerobic conditions to 1 and 10 microg/ml under 21% oxygen in the absence and presence of carbon dioxide, respectively. This study demonstrates the controlling effects of both oxygen and carbon dioxide on TSST-1 production.