The fate of mutations surfing on the wave of a range expansion.
Many species, including humans, have dramatically expanded their range in the past, and such range expansions had certainly an impact on their genetic diversity. For example, mutations arising in populations at the edge of a range expansion can sometimes surf on the wave of advance and thus reach a larger spatial distribution and a much higher frequency than would be expected in stationary populations. We study here this surfing phenomenon in more detail, by performing extensive computer simulations under a two-dimensional stepping-stone model. We find that the probability of survival of a new mutation depends to a large degree on its proximity to the edge of the wave. Demographic factors such as deme size, migration rate, and local growth rate also influence the fate of these new mutations. We also find that the final spatial and frequency distributions depend on the local deme size of a subdivided population. This latter result is discussed in the light of human expansions in Europe as it should allow one to distinguish between mutations having spread with Paleolithic or Neolithic expansions. By favoring the spread of new mutations, a consequence of the surfing phenomenon is to increase the rate of evolution of spatially expanding populations.


Total respiratory impedance measured by means of the forced oscillation technique in subjects with and without respiratory complaints.
The purpose of this study was to determine whether the forced oscillation technique is more sensitive than spirometry to detect lung function alterations in subjects with respiratory complaints. The input impedance of the respiratory system (between 2 and 24 Hz) and maximal expiratory flows and volumes were measured in 1,255 subjects referred for routine spirometry. A questionnaire concerning respiratory complaints was administered. A discriminant analysis was performed between subgroups of subjects without (137 males and 140 females), with moderate (115 males and 109 females) and with marked respiratory complaints (149 males and 132 females). A clear-cut separation was achieved by this analysis only between those subjects without and with marked complaints. Both lung volumes and flows as well as impedance parameters (mean value and frequency dependence of resistance in females, mean resistance in males) contributed to the discrimination of subjects without and with marked respiratory complaints, although there was only a moderate decrease of discriminative power when the impedance parameters were excluded. The contribution of the forced oscillation parameters to discriminative power was larger in females than in males (40 vs 19%), which may be related to the higher prevalence of asthma in our population of females. Excluding the subjects with marked functional impairment improved the share of forced oscillation parameters only slightly with respect to lung volumes and flows (females 54 vs males 23%). Considered separately, however, the sensitivity of spirometry and forced oscillation technique to detect symptomatic people appeared to be similar. We conclude that impedance measurements by forced oscillation technique and routine spirometry are both associated with respiratory complaints. Our results indicate that the information provided by impedance measurements can be complimentary to that obtained by spirometric indices.


Rescue of Infectious Sindbis Virus by Yeast Spheroplast-Mammalian Cell Fusion.
Sindbis virus (SINV), a positive-sense single stranded RNA virus that causes mild symptoms in humans, is transmitted by mosquito bites. SINV reverse genetics have many implications, not only in understanding alphavirus transmission, replication cycle, and virus-host interactions, but also in biotechnology and biomedical applications. The rescue of SINV infectious particles is usually achieved by transfecting susceptible cells (BHK-21) with SINV-infectious mRNA genomes generated from cDNA constructed via in vitro translation (IVT). That procedure is time consuming, costly, and relies heavily on reagent quality. Here, we constructed a novel infectious SINV cDNA construct that expresses its genomic RNA in yeast cells controlled by galactose induction. Using spheroplasts made from this yeast, we established a robust polyethylene glycol-mediated yeast: BHK-21 fusion protocol to rescue infectious SINV particles. Our approach is timesaving and utilizes common lab reagents for SINV rescue. It could be a useful tool for the rescue of large single strand RNA viruses, such as SARS-CoV-2.


Effects of supplementation with n-3 polyunsaturated fatty acids on cognitive performance and cardiometabolic risk markers in healthy 51 to 72 years old subjects: a randomized controlled cross-over study.
Higher plasma n-3 polyunsaturated fatty acids (PUFA) have been associated with a lower risk of age related cognitive decline, and to beneficially affect cardiometabolic risk factors. A relation exists between metabolic disorders such as diabetes type 2 and cognitive decline. Results regarding the potential effects of n-3 PUFA on risk factors in healthy subjects are divergent, and studies regarding the possible relation between cardiometabolic parameters and cognitive performance are scarce. The objective was to evaluate the effects of five weeks intake of long chain n-3 PUFA on cognitive performance in healthy individuals, and to exploit the possible relation between outcomes in cognitive tests to cardiometabolic risk parameters. Fish oil n-3 PUFA (3g daily) were consumed during 5 weeks separated by a 5 week washout period in a cross-over placebo controlled study, including 40 healthy middle aged to elderly subjects. Cognitive performance was determined by tests measuring working memory (WM) and selective attention. Supplementation with n-3 PUFA resulted in better performance in the WM-test compared with placebo (p < 0.05). In contrast to placebo, n-3 PUFA lowered plasma triacylglycerides (P < 0.05) and systolic blood pressure (p < 0.0001). Systolic blood pressure (p < 0.05), f-glucose (p = 0.05), and s-TNF-α (p = 0.05), were inversely related to the performance in cognitive tests. Intake of n-3 PUFA improved cognitive performance in healthy subjects after five weeks compared with placebo. In addition, inverse relations were obtained between cardiometabolic risk factors and cognitive performance, indicating a potential of dietary prevention strategies to delay onset of metabolic disorders and associated cognitive decline.


Localization of steroidogenic enzymes in macaque luteal tissue during the menstrual cycle and simulated early pregnancy: immunohistochemical evidence supporting the two-cell model for estrogen production in the primate corpus luteum.
It is hypothesized that the two-cell model for estrogen production by the ovarian follicle is preserved in the primate corpus luteum, but there is little direct evidence to support this theory. To determine the sites of androgen and estrogen synthesis within the primate corpus luteum and to ascertain whether changes in steroid hormone levels are related to steroidogenic enzyme expression, the enzymes converting progesterone to androgen (cytochrome P450 17alpha-hydroxylase/17,20 lyase; P450(c17)) and then to estrogen (aromatase; P450(arom)), as well as P450 side-chain cleavage (P450(scc)) and 3beta-hydroxysteroid dehydrogenase (3beta HSD), were detected by immunohistochemistry in macaque luteal tissue throughout the menstrual cycle and simulated early pregnancy. Corpora lutea were collected from rhesus monkeys in the early (Days 2-4 post-LH surge), mid (Days 6-8), mid-late (Days 10-12), and late (Days 14-15) luteal phase and after 1, 3, 6, or 9 days of hCG treatment that began on Day 9 of the luteal phase. Specific cytoplasmic staining for P450(c17), P450(arom), P450(scc), and 3beta HSD was present in luteal cells, but not in the microvasculature, within all luteal tissues examined. P450(c17)-stained luteal cells were located along the vascular tracts and around the periphery of the corpus luteum. Intensely stained luteal cells were associated with blood vessels entering from the outer surface of the corpus luteum, but not with blood vessels returning from the connective tissue centrum. In contrast, P450(arom)-stained luteal cells were distributed throughout the luteal parenchyma. P450(c17) staining intensity was similar at all stages of the luteal phase; however, the number and intensity of P450(arom)-stained cells decreased by late luteal phase. In simulated early pregnancy, cells stained for P450(c17) were present near blood vessels, with some positive cells scattered throughout the corpus luteum. P450(arom) immunostaining was heterogeneous within the corpus luteum; many intensely stained cells were interspersed among others that were only lightly stained. Overall, cellular staining for P450(c17) and P450(arom) remained intense through 9 days of simulated early pregnancy. In contrast, P450(scc) and 3beta HSD immunoreactivity were not located in distinct luteal compartments. These results are consistent with a two-cell model for steroid hormone production in the primate corpus luteum, whereby paraluteal (theca-luteal) cells produce androgen substrate that is converted to estrogens by true (granulosa-) luteal cells. The divergence in enzyme detection as the luteal phase progresses, with P450(c17) labeling high and P450(arom) staining having decreased, suggests a shift in the function of the corpus luteum as it ages. Enzyme localization during chorionic gonadotropin exposure simulating early pregnancy demonstrates the continued capacity of the primate corpus luteum to produce steroid hormones.


Identification of active components in Yixinshu Capsule with protective effects against myocardial dysfunction on human induced pluripotent stem cell-derived cardiomyocytes by an integrative approach.
Traditional Chinese medicine (TCM) preparations have significant effects on some refractory diseases; however, these compositions are complex and their mechanisms are unknown. Identification of the active components in these preparations is essential. The mortality rate for heart failure (HF) has been increasing in recent years, and myocardial dysfunction (MD) has been proved to be the pathological basis of HF. Yixinshu Capsule (YXSC) is a multi-component oral drug with therapeutic effects on HF. However, the key active components are still unclear. In this study, YXSC intestinal absorption liquid (IAL) was used and 62 compounds were identified by an analytical chemistry approach. Then, a compound - target - function network was established with a bioinformatics analysis tool. Finally, a cell model of MD on human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) was used to verify the therapeutic effects of the active components of YXSC. Schisandrin A (Sch A) and schisandrin B (Sch B) were demonstrated to be the active components of YXSC by attenuating endothelin-1 (ET-1)-induced contraction dysfunction, brain natriuretic peptide (BNP) content elevation, and the morphological changes of hiPS-CMs. For the first time, our data illustrate the potent protective effects of Sch A and Sch B on ET-1-induced dysfunctional hiPS-CMs and revealed their effective targets and pathways. The integrative approach used in our study was applied to identify active components in TCM preparations and excavate the possible mechanisms.


First Report of Fusarium pseudograminearum Causing Crown Rot of Wheat in Henan, China.
Fusarium pseudograminearum (O'Donnell & Aoki), a residue-borne pathogen, is responsible for crown rot of wheat (Triticum aestivum L.). Since its first detection in Queensland, Australia in 1951, it has been reported in many other countries, but not China (2). In May 2011, a crown rot disease was observed in wheat cv. Aikang 58 in a wheat-maize rotation, irrigable and loam field in Henan Province, China. Diseased wheat plants showed honey brown discoloration in the stem bases and whitehead in some plants, which are symptoms of crown rot with about 70% incidence in a surveyed field (2). The pathogen was isolated from diseased stem base on potato dextrose agar (PDA) after being surface-disinfested with 5% NaClO solution for 2 min. Pure cultures were established on carnation leaf agar (CLA) through a single spore technique and identified by morphological and molecular methods according to protocols described previously (1,3,4). Macroconidia of F. pseudograminearum were formed in abundant sporodochia on CLA cultures grown under the BLB light. Macroconidia were usually five septate (about three to seven) and 27 to 91 × 2.7 to 5.5 μm. Colonies grown on PDA from a single conidium in the dark at 25°C had average radial growth rates of ~4.7 to 9.9 mm per day. Colony pigment on PDA grown under light varied from rose to burgundy, while mycelium ranged from rose to yellow white. Two isolates (WZ-8A and WZ-2B) were selected for molecular identification. The translation elongation factor 1-α gene and rDNA ITS gene were amplified by PCR using the specific primers described previously (4). PCR products were sequenced (GenBank Accession Nos. JN862232 to JN862235). Phylogenic analysis of the sequence indicated that the isolates were identified as F. pseudograminearum. The identification was further confirmed by the F. pseudograminearum species-specific PCR primers (Fp1-1: CGGGGTAGTTTCACATTTCCG and Fp1-2: GAGAATGTGATGACGACAATA) (1). The expected PCR products of 520 bp were produced only in F. pseudograminearum. Isolates WZ-2B and WZ-8A were deposited in the Agriculture Culture Collection of China as ACCC38067 and ACCC 38068, respectively. Pathogenicity tests were conducted by inoculating winter wheat cultivar Wenmai 19 with isolates WZ-8A and WZ-2B through soil inoculation. Inoculum was prepared by growing cultures on sterilized wheat bran and chopped wheat-straw (4:1, v/v) after incubation at 25°C for 2 weeks. This inoculum was added to sterilized soil at 1% by volume and no inoculum was added in control treatment. Five seeds were planted in a 15 cm wide pot in a 20 to 25°C greenhouse, with six replications. Seedling death and crown browning occurred in the inoculated wheat plants after 4 weeks with over 90% incidence, while no symptoms developed in the control plants. The fungus was reisolated from inoculated plants, fulfilling Koch's postulates. To our knowledge, this is the first report of F. pseudograminearum causing crown rot of wheat in China. Considering Henan is the largest wheat production province in China with over 5 million hectares planting area, and the soil and climate conditions are suitable for this disease, it will be a important pathogen of wheat in Henan in the future. References: (1) T. Aoki et al. Mycologia 91:597, 1999. (2) L. W. Burgess. Page 271 in: Crown Rot of Wheat: Fusarium. B. A. Summerell et al., eds. APS Press, St. Paul, MN, 2001. (3) R. G. Francis et al. Trans. Brit. Mycol. Soc. 68:421, 1977. (4) J. B. Scott et al. Mycol. Res. 110:1413, 2006.


Social justice and research using human biological material: A response to Mahomed, Nöthling-Slabbert and Pepper.
Social justice in the context of research using human biological material is an important contemporary legal-ethical issue. A question at the heart of this issue is the following: Is it fair to expect a research participant (a person who participates in such research by, among others, making available biological material from his or her body) to participate on an altruistic basis, while the researchers and the investors in the research can gain commercially from the research? In a recent article, Mahomed, Nöthling-Slabbert and Pepper proposed that research participants should be entitled to share in the profits emanating from such research via a proposed new statutory right to the intellectual property emanating from such research. In order to stimulate debate on this important issue of social justice, this article responds to the position of Mahomed et al. by focusing on two main points: Firstly, I contend that Mahomed et al. fail to make a convincing argument in favour of shifting away from altruism; secondly, I caution against framing the debate in terms of the binary poles of altruism v. profitsharing, and suggest that should healthcare public policy ever move away from altruism, various non-monetary forms of benefit-sharing by research participants should be considered.


Genetic Risk Can Be Decreased: Quitting Smoking Decreases and Delays Lung Cancer for Smokers With High and Low CHRNA5 Risk Genotypes - A Meta-Analysis.
Recent meta-analyses show that individuals with high risk variants in CHRNA5 on chromosome 15q25 are likely to develop lung cancer earlier than those with low-risk genotypes. The same high-risk genetic variants also predict nicotine dependence and delayed smoking cessation. It is unclear whether smoking cessation confers the same benefits in terms of lung cancer risk reduction for those who possess CHRNA5 risk variants versus those who do not. Meta-analyses examined the association between smoking cessation and lung cancer risk in 15 studies of individuals with European ancestry who possessed varying rs16969968 genotypes (N=12,690 ever smokers, including 6988 cases of lung cancer and 5702 controls) in the International Lung Cancer Consortium. Smoking cessation (former vs. current smokers) was associated with a lower likelihood of lung cancer (OR=0.48, 95%CI=0.30-0.75, p=0.0015). Among lung cancer patients, smoking cessation was associated with a 7-year delay in median age of lung cancer diagnosis (HR=0.68, 95%CI=0.61-0.77, p=4.9∗10-10). The CHRNA5 rs16969968 risk genotype (AA) was associated with increased risk and earlier diagnosis for lung cancer, but the beneficial effects of smoking cessation were very similar in those with and without the risk genotype. We demonstrate that quitting smoking is highly beneficial in reducing lung cancer risks for smokers regardless of their CHRNA5 rs16969968 genetic risk status. Smokers with high-risk CHRNA5 genotypes, on average, can largely eliminate their elevated genetic risk for lung cancer by quitting smoking- cutting their risk of lung cancer in half and delaying its onset by 7years for those who develop it. These results: 1) underscore the potential value of smoking cessation for all smokers, 2) suggest that CHRNA5 rs16969968 genotype affects lung cancer diagnosis through its effects on smoking, and 3) have potential value for framing preventive interventions for those who smoke.


Growth and C allocation of Populus tremuloides genotypes in response to atmospheric CO2 and soil N availability.
We grew cuttings of two early (mid Oct.) and two late (early Nov.) leaf-fall Populus tremuloides Michx. genotypes (referred to as genotype pairs) for c. 150 d in open-top chambers to understand how twice-ambient (elevated) CO2 and soil N availability would affect growth and C allocation. For the study, we selected genotypes differing in leaf area duration to find out if late-season photosynthesis influenced C allocation to roots. Both elevated CO2 and high soil N availability significantly increased estimated whole-tree photosynthesis, but they did so in different ways. Elevated CO2 stimulated leaf-level photosynthesis rates, whereas high soil N availability resulted in greater total plant leaf area. The early leaf-fall genotype pair had significantly higher photosynthesis rates per unit leaf area than the late leaf-fall genotype pair and elevated CO2 enhanced this difference. The early leaf-fall genotype pair had less leaf area than the late leaf-fall genotype pair, and their rate of leaf area development decreased earlier in the season. Across both genotype pairs, high soil N availability significantly increased fine root length production and mortality by increasing both the amount of root length present, and by decreasing the life span of individual roots. Elevated CO2 resulted in significantly increased fine root production and mortality in high N but not low N soil and did not affect fine root life span. The early leaf-fall genotype pair had significantly greater fine root length production than the late leaf-fall genotype pair across all CO2 and N treatments. These differences in belowground C allocations are consistent with the hypothesis that belowground C and N cycling is strongly influenced by soil N availability and will increase under elevated atmospheric CO2 . In addition, this study reinforces the need for better understanding of the variation in tree responses to elevated CO2 , within and among species.


Coronary Injury Score Correlates with Proliferating Cells and Alpha-Smooth Muscle Actin Expression in Stented Porcine Coronary Arteries.
Neointimal formation and cell proliferation resulting into in-stent restenosis is a major pathophysiological event following the deployment of stents in the coronary arteries. In this study, we assessed the degree of injury, based on damage to internal elastic lamina, media, external elastic lamina, and adventitia following the intravascular stenting, and its relationship with the degree of smooth muscle cell proliferation. We examined the smooth muscle cell proliferation and their phenotype at different levels of stent injury in the coronary arteries of domestic swine fed a normal swine diet. Five weeks after stent implantation, swine with and without stents were euthanized and coronaries were excised. Arteries were embedded in methyl methacrylate and sections were stained with H&E, trichrome, and Movat's pentachrome. The expression of Ki67, α-smooth muscle actin (SMA), vimentin, and HMGB1 was evaluated by immunofluorescence. There was a positive correlation between percent area stenosis and injury score. The distribution of SMA and vimentin was correlated with the degree of arterial injury such that arteries that had an injury score >2 did not have immunoreactivity to SMA in the neointimal cells near the stent struts, but these neointimal cells were positive for vimentin, suggesting a change in the smooth muscle cell phenotype. The Ki67 and HMGB1 immunoreactivity was highly correlated with the fragmentation of the IEL and injury in the tunica media. Thus, the extent of coronary arterial injury during interventional procedure will dictate the degree of neointimal hyperplasia, in-stent restenosis, and smooth muscle cell phenotype.


Surfactant-assisted synthesis of ceria-titania-rich mesoporous silica materials and their catalytic activity towards photodegradation of organic dyes.
Ceria-titania doped highly ordered hexagonal and cubic mesoporous silica composite samples with varying amounts of Ce and Ti have been synthesized using hexadecyltrimethylammonium bromide (CTAB) surfactant as a template under basic conditions in the presence of ammonia. The removal of the template resulted in the formation of mesoporous composites doped with Ce and Ti, which were thoroughly characterized via small-angle X-ray scattering (SAXS), wide-angle X-ray diffraction (WAXRD), N2 adsorption, scanning and transmission electron microscopy analysis with energy-dispersive spectrometry mapping (SEM-TEM-EDS), inductively coupled plasma atomic emission spectrophotometry (ICP-AES), and ultraviolet-visible diffuse reflectance spectrometry (UV-visible). The electronic states of Ti and Ce species present on the surface of silica were also investigated by X-ray photoelectron spectroscopy (XPS). The catalytic activity of these Ti-modified ceria-silica samples for the photodecomposition of rhodamine B under visible-light irradiation was investigated. The results revealed that a sample with a higher Ti content, as well as equal amounts of Ce and Ti, displayed higher catalytic activity for the photodegradation of the organic dye rhodamine B.


Emergency carotid artery surgery in neurologically unstable patients.
Although angiography and carotid artery surgery are illadvised in patients with acute, profound stroke, there is no consensus on the management of patients with stroke in evolution, waxing and waning neurologic deficits, or crescendo transient ischemic attacks. This type of clinical picture was associated with a critical, unstable lesion of the internal carotid artery in each of 12 patients. Emergency angiography permitted identification of the lesions responsible for the varying neurologic manifestations, and emergency carotid thromboendarterectomy produced prompt, complete recovery in all but one patient, who had a total carotid occlusion, received no operation, and died of a cerebral infarction. Based on our experience with these 12 patients, an aggressive diagnostic and therapeutic approach is recommended for patients with acute unstable cerebrovascular disease.


Common misconceptions in the diagnosis and management of anemia in inflammatory bowel disease.
Anemia is the most common systemic complication of inflammatory bowel disease (IBD); so common that it is almost invariably not investigated and rarely treated. Several misconceptions are the reason for these clinical errors, and our goal will be to review them. The most common misconceptions are: anemia is uncommon in IBD; iron deficiency is also uncommon; just by treating the intestinal disease, anemia will be corrected; iron deficiency is the only cause for anemia in IBD; ferritin is an accurate parameter for the diagnosis of iron deficiency in IBD; the impact of anemia on the quality of life of IBD patients is limited; iron supplementation is rarely needed in IBD; high-dose oral iron solves the problem of iron malabsorption in IBD; intravenous (IV) iron is dangerous and of no proven benefit in IBD; IV iron is useful only for severe anemia; and erythropoietin has no role in the treatment of IBD anemia. These misconceptions are not evidence-based. On the contrary, there is enough evidence to support the following statements: (a) anemia is very common in IBD, (b) anemia should be investigated with care because many factors can be responsible, (c) treatment of anemia results in clear improvement in the objective parameters of well-being, especially in the quality of life, (d) IV iron is safe and effective in the treatment of iron deficiency anemia in IBD patients, and (e) erythropoietin is useful in a subset of patients with refractory anemia. Anemia diagnosis and treatment must not be neglected in IBD patients, and several misconceptions should be promptly abandoned.


When to Suspect a Diagnosis of Amyloidosis.
Amyloidosis is a group of complex diseases caused by extracellular deposition of pathological insoluble fibrillary protein in organs and tissues and may result in severe organ dysfunction. Despite the etiological heterogeneity of systemic amyloidosis, the clinical manifestations of the different forms of amyloidosis largely overlap and depend upon the effected organ. The signs and symptoms that should raise suspicion for the potential diagnosis of amyloidosis are usually nonspecific; therefore, establishing the diagnosis is difficult, and early diagnosis requires clinical suspicion. Light chain (AL) amyloidosis may present with highly specific signs such as macroglossia and periorbital purpura, but these signs are insensitive. Amyloidosis is still underdiagnosed, even though treatments are now available and are effective in improving patient's survival and quality of life. Cardiac amyloidosis is the major determinant of survival, and the earlier it is detected the better the survival. All MGUS patients should be routinely screened for AL amyloidosis by a focused history and physical examination and routine assessment of urine albumin. The aim of this review is to provide clinicians with knowledge about the signs and symptoms that raise the suspicion of amyloidosis, bearing in mind the importance of early diagnosis of this disease.


Nuclear localization of mouse fibroblast growth factor 2 requires N-terminal and C-terminal sequences.
In vertebrates, different isoforms of fibroblast growth factor 2 (FGF2) exist, which differ by their N-terminal extension. They show different localization and expression levels and exert distinct biological effects. Nevertheless, genetic inactivation of all FGF2 isoforms in the mouse results in only mild phenotypes. Here, we analyzed mouse FGF2, and show that, as in the human, mouse FGF2 contains CTG-initiated high molecular-weight (HMW) isoforms, which contain a nuclear localization signal, and which mediate localization of this isoform to the nucleus. Using green fluorescent protein-FGF2 fusions, we furthermore observed, that C-terminal deletions disable nuclear localization of the short low-molecular-weight (LMW) 18-kDa isoform. This loss of specific localization is accompanied by a loss in heparin binding. We therefore suggest that, first, localization of mouse FGF2 is comparable to that in other vertebrates and, second, FGF2 contains at least two sequences important for nuclear localization, a nuclear localization sequence at the N terminus which is only contained in the HMW isoform, and another sequence at the C terminus, which is only required for localization of the LMW 18-kDa isoform.


Extreme positive allometry of animal adhesive pads and the size limits of adhesion-based climbing.
Organismal functions are size-dependent whenever body surfaces supply body volumes. Larger organisms can develop strongly folded internal surfaces for enhanced diffusion, but in many cases areas cannot be folded so that their enlargement is constrained by anatomy, presenting a problem for larger animals. Here, we study the allometry of adhesive pad area in 225 climbing animal species, covering more than seven orders of magnitude in weight. Across all taxa, adhesive pad area showed extreme positive allometry and scaled with weight, implying a 200-fold increase of relative pad area from mites to geckos. However, allometric scaling coefficients for pad area systematically decreased with taxonomic level and were close to isometry when evolutionary history was accounted for, indicating that the substantial anatomical changes required to achieve this increase in relative pad area are limited by phylogenetic constraints. Using a comparative phylogenetic approach, we found that the departure from isometry is almost exclusively caused by large differences in size-corrected pad area between arthropods and vertebrates. To mitigate the expected decrease of weight-specific adhesion within closely related taxa where pad area scaled close to isometry, data for several taxa suggest that the pads' adhesive strength increased for larger animals. The combination of adjustments in relative pad area for distantly related taxa and changes in adhesive strength for closely related groups helps explain how climbing with adhesive pads has evolved in animals varying over seven orders of magnitude in body weight. Our results illustrate the size limits of adhesion-based climbing, with profound implications for large-scale bio-inspired adhesives.


Intact Transition Epitope Mapping - Targeted High-Energy Rupture of Extracted Epitopes (ITEM-THREE).
Epitope mapping, which is the identification of antigenic determinants, is essential for the design of novel antibody-based therapeutics and diagnostic tools. ITEM-THREE is a mass spectrometry-based epitope mapping method that can identify epitopes on antigens upon generating an immune complex in electrospray-compatible solutions by adding an antibody of interest to a mixture of peptides from which at least one holds the antibody's epitope. This mixture is nano-electrosprayed without purification. Identification of the epitope peptide is performed within a mass spectrometer that provides an ion mobility cell sandwiched in-between two collision cells and where this ion manipulation setup is flanked by a quadrupole mass analyzer on one side and a time-of-flight mass analyzer on the other side. In a stepwise fashion, immune-complex ions are separated from unbound peptide ions and dissociated to release epitope peptide ions. Immune complex-released peptide ions are separated from antibody ions and fragmented by collision induced dissociation. Epitope-containing peptide fragment ions are recorded, and mass lists are submitted to unsupervised data base search thereby retrieving both, the amino acid sequence of the epitope peptide and the originating antigen. ITEM-THREE was developed with antiTRIM21 and antiRA33 antibodies for which the epitopes were known, subjecting them to mixtures of synthetic peptides of which one contained the respective epitope. ITEM-THREE was then successfully tested with an enzymatic digest of His-tagged recombinant human β-actin and an antiHis-tag antibody, as well as with an enzymatic digest of recombinant human TNFα and an antiTNFα antibody whose epitope was previously unknown.


Font adaptive word indexing of modern printed documents.
We propose an approach for the word-level indexing of modern printed documents which are difficult to recognize using current OCR engines. By means of word-level indexing, it is possible to retrieve the position of words in a document, enabling queries involving proximity of terms. Web search engines implement this kind of indexing, allowing users to retrieve Web pages on the basis of their textual content. Nowadays, digital libraries hold collections of digitized documents that can be retrieved either by browsing the document images or relying on appropriate metadata assembled by domain experts. Word indexing tools would therefore increase the access to these collections. The proposed system is designed to index homogeneous document collections by automatically adapting to different languages and font styles without relying on OCR engines for character recognition. The approach is based on three main ideas: the use of Self Organizing Maps (SOM) to perform unsupervised character clustering, the definition of one suitable vector-based word representation whose size depends on the word aspect-ratio, and the run-time alignment of the query word with indexed words to deal with broken and touching characters. The most appropriate applications are for processing modern printed documents (17th to 19th centuries) where current OCR engines are less accurate. Our experimental analysis addresses six data sets containing documents ranging from books of the 17th century to contemporary journals.


[Climate change influences the incidence of arthropod-borne diseases in the Netherlands].
Climate change is associated with changes in the occurrence of arthropod-borne diseases. It is difficult to foresee which arthropod-borne diseases will appear in the Netherlands due to climate change. Climate change influences the prevalence of ticks and may lead to a further increase in Lyme disease and an increased risk of the introduction of rickettsioses. With further warming of the climate there is a real possibility of settlement of the mosquito Aedes albopictus and introduction of the sandfly in the Netherlands. Whether this will lead to circulation of micro-organisms transmitted by these vectors (e.g. West Nile virus, Dengue virus, Leishmania) is not clear. Continued vigilance is necessary, even for vector-borne diseases that appear to be less relevant for the Netherlands.


Design and development of a novel fused filament fabrication (FFF) 3D printed diffusion cell with UV imaging capabilities to characterise permeation in pharmaceutical formulations.
The present work aimed at designing and developing a novel 3D printed diffusion cell capable of UV imaging using the fused filament fabrication (FFF) method. UV imaging has proven to be very versatile in the area of pharmaceutics giving insights into various phenomena including the dissolution behaviour of dosage forms, intrinsic dissolution rates and the drug precipitation processes. A 3D printed diffusion cell in the similitude of a Franz cell was successfully printed using polylactic acid (PLA) filaments equipped with quartz for the imaging area. A model ibuprofen (IBU) gel formulation was tested by introducing the dosage form through the 3D printed donor compartment. The drug concentration permeated through the skin mimic (silicone membrane) was determined from the 3D printed receptor compartment using UV imaging in real-time. The results showed successful UV imaging of the permeation of IBU gel in the novel diffusion cell potentially negating further analytical testing such as the HPLC process required for Franz cell tests thereby reducing costs. Potential interactions between the drug and filament used in the 3D printed process suggests although this concept can be moved towards commercialisation, care should be taken with choice of filament used in the 3D printing process.


Alternative mRNA splicing produces a novel biologically active short isoform of PGC-1alpha.
The transcriptional co-activator PGC-1alpha regulates functional plasticity in adipose tissue by linking sympathetic input to the transcriptional program of adaptive thermogenesis. We report here a novel truncated form of PGC-1alpha (NT-PGC-1alpha) produced by alternative 3' splicing that introduces an in-frame stop codon into PGC-1alpha mRNA. The expressed protein includes the first 267 amino acids of PGC-1alpha and 3 additional amino acids from the splicing insert. NT-PGC-1alpha contains the transactivation and nuclear receptor interaction domains but is missing key domains involved in nuclear localization, interaction with other transcription factors, and protein degradation. Expression and subcellular localization of NT-PGC-1alpha are dynamically regulated in the context of physiological signals that regulate full-length PGC-1alpha, but the truncated domain structure conveys unique properties with respect to protein-protein interactions, protein stability, and recruitment to target gene promoters. Therefore, NT-PGC-1alpha is a co-expressed, previously unrecognized form of PGC-1alpha with functions that are both unique from and complementary to PGC-1alpha.


How the Eukaryotic Replisome Achieves Rapid and Efficient DNA Replication.
The eukaryotic replisome is a molecular machine that coordinates the Cdc45-MCM-GINS (CMG) replicative DNA helicase with DNA polymerases α, δ, and ε and other proteins to copy the leading- and lagging-strand templates at rates between 1 and 2 kb min-1. We have now reconstituted this sophisticated machine with purified proteins, beginning with regulated CMG assembly and activation. We show that replisome-associated factors Mrc1 and Csm3/Tof1 are crucial for in vivo rates of replisome progression. Additionally, maximal rates only occur when DNA polymerase ε catalyzes leading-strand synthesis together with its processivity factor PCNA. DNA polymerase δ can support leading-strand synthesis, but at slower rates. DNA polymerase δ is required for lagging-strand synthesis, but surprisingly also plays a role in establishing leading-strand synthesis, before DNA polymerase ε engagement. We propose that switching between these DNA polymerases also contributes to leading-strand synthesis under conditions of replicative stress.


An Interpretable Classification Model Using Gluten-Specific TCR Sequences Shows Diagnostic Potential in Coeliac Disease.
Coeliac disease (CeD) is a T-cell mediated enteropathy triggered by dietary gluten which remains substantially under-diagnosed around the world. The diagnostic gold-standard requires histological assessment of intestinal biopsies taken at endoscopy while consuming a gluten-containing diet. However, there is a lack of concordance between pathologists in histological assessment, and both endoscopy and gluten challenge are burdensome and unpleasant for patients. Identification of gluten-specific T-cell receptors (TCRs) in the TCR repertoire could provide a less subjective diagnostic test, and potentially remove the need to consume gluten. We review published gluten-specific TCR sequences, and develop an interpretable machine learning model to investigate their diagnostic potential. To investigate this, we sequenced the TCR repertoires of mucosal CD4+ T cells from 20 patients with and without CeD. These data were used as a training dataset to develop the model, then an independently published dataset of 20 patients was used as the testing dataset. We determined that this model has a training accuracy of 100% and testing accuracy of 80% for the diagnosis of CeD, including in patients on a gluten-free diet (GFD). We identified 20 CD4+ TCR sequences with the highest diagnostic potential for CeD. The sequences identified here have the potential to provide an objective diagnostic test for CeD, which does not require the consumption of gluten.


Experimental phage therapy of burn wound infection: difficult first steps.
Antibiotic resistance has become a major public health problem and the antibiotics pipeline is running dry. Bacteriophages (phages) may offer an 'innovative' means of infection treatment, which can be combined or alternated with antibiotic therapy and may enhance our abilities to treat bacterial infections successfully. Today, in the Queen Astrid Military Hospital, phage therapy is increasingly considered as part of a salvage therapy for patients in therapeutic dead end, particularly those with multidrug resistant infections. We describe the application of a well-defined and quality controlled phage cocktail, active against Pseudomonas aeruginosa and Staphylococcus aureus, on colonized burn wounds within a modest clinical trial (nine patients, 10 applications), which was approved by a leading Belgian Medical Ethical Committee. No adverse events, clinical abnormalities or changes in laboratory test results that could be related to the application of phages were observed. Unfortunately, this very prudent 'clinical trial' did not allow for an adequate evaluation of the efficacy of the phage cocktail. Nevertheless, this first 'baby step' revealed several pitfalls and lessons for future experimental phage therapy and helped overcome the psychological hurdles that existed to the use of viruses in the treatment of patients in our burn unit.


Epidermal growth factor receptor mutations in colorectal cancer patients.
The epidermal growth factor receptor (EGFR) plays an important role in tumorigenesis and tumor progression of colorectal cancer and leads to the activation of intracellular signaling pathways. The use of anti-EGFR-targeted therapy has increased for patients with colorectal cancer, but patients with EGFR mutations will be resistant to anti-EGFR-targeted therapy. The identification of gene mutations is critical in cancer treatment; therefore, the aim of this study is to investigate the incidences of EGFR mutations in colorectal cancer patients in Korea. We retrospectively reviewed 58 colorectal cancer patients who underwent surgery between 2003 and 2006. We analyzed their EGFR mutations in four loci by DNA sequencing. In addition, we analyzed the correlation between the presence of EGFR mutation and patients' clinicopathologic features. Of the 58 patients, 35 patients were male and 23 were female. Their mean age was 63.28 ± 11.18 years. Two patients (3.45%) were diagnosed as stage Tis, 7 patients (12.07%) as stage I, 24 patients (41.38%) as stage II, 20 patients (34.48%) as stage III, and 5 patients (8.62%) as stage IV. As a result of mutational analysis, EGFR mutations on exon 20 were detected in 13 patients (22.41%, G→A transitions). No EGFR mutations were detected on exons 18, 19, and 21. EGFR mutation was increased in the earlier stage and in the absence of lymph node metastasis (P = 0.028). The incidence of EGFR mutation in Korean colorectal cancer patients is 22.41%. In addition, EGFR mutation was significantly increased in the earlier stage and in the absence of lymph node metastasis.


Association Between Visceral Fat Accumulation and Exercise Tolerance in Non-Obese Subjects Without Diabetes.
We examined the associations between visceral fat accumulation, presence of the components of metabolic syndrome (MetS), and exercise tolerance in non-obese subjects without diabetes. Seventy-four non-obese, non-diabetic Japanese men were enrolled. The subjects were divided into the following two groups: non-obese subjects without any MetS risk factors (n = 38, Group A) and non-obese subjects with one or two MetS risk factors (n = 36, Group B). Anthropometric and metabolic parameters were measured. The response of heart rate (HR) and blood pressure (BP), and exercise tolerance were also evaluated with a cardiopulmonary exercise test using a bicycle ergometer. The body mass index, abdominal circumference, visceral fat area, and homeostasis model assessment-insulin resistance, were significantly higher, while levels of anaerobic threshold and maximal oxygen uptake were significantly lower in Group B than in Group A. The levels of resting HR, resting BP, and BP at maximal exercise were significantly higher in Group B than in Group A. There were no significant differences in the HR at maximal exercise as well as the HR and BP after exercise between the two groups. The visceral fat area was significantly and negatively correlated with exercise tolerance. Multivariate linear regression analyses demonstrated that visceral fat area, but not abdominal circumference, was significantly and independently associated with maximal oxygen uptake. These data suggest that the visceral fat area is a significant determinant for exercise tolerance even in non-obese subjects without diabetes.


Large-scale analysis of SARS-CoV-2 synonymous mutations reveals the adaptation to the human codon usage during the virus evolution.
Many large national and transnational studies have been dedicated to the analysis of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) genome, most of which focused on missense and nonsense mutations. However, approximately 30 per cent of the SARS-CoV-2 variants are synonymous, therefore changing the target codon without affecting the corresponding protein sequence. By performing a large-scale analysis of sequencing data generated from almost 400,000 SARS-CoV-2 samples, we show that silent mutations increasing the similarity of viral codons to the human ones tend to fixate in the viral genome overtime. This indicates that SARS-CoV-2 codon usage is adapting to the human host, likely improving its effectiveness in using the human aminoacyl-tRNA set through the accumulation of deceitfully neutral silent mutations. One-Sentence Summary. Synonymous SARS-CoV-2 mutations related to the activity of different mutational processes may positively impact viral evolution by increasing its adaptation to the human codon usage.


Brain-derived neurotrophic factor supports the survival of cultured rat retinal ganglion cells.
Brain-derived neurotrophic factor (BDNF) is a small, basic protein purified from the mammalian brain that has been shown previously to support the survival of cultured spinal sensory neurons (Barde et al., 1982). In current studies, BDNF was tested for its ability to support the survival of cultured CNS cells isolated from the perinatal rat retina. Both immunofluorescent labeling of Thy-1 and prior retrograde labeling with HRP were used as retinal ganglion cell markers in vitro. With embryonic day (E) 17 retinas, it was found that BDNF allowed the survival of a small subpopulation of neurons (about 7% of the cells plated at this age) identified by the immunofluorescent labeling of Thy-1. No detectable effects were seen when either the total number of cells or the number of tetanus toxin-positive neurons was measured. BDNF also had an effect on cultured neurons retrogradely labeled after HRP injections in the superior colliculi of neonatal rats. The BDNF-responsive population was therefore detected only in retinal cultures with specific markers and identified as consisting of retinal ganglion cells. These cells could be enriched about 80-fold by density gradient centrifugation, and purified ganglion cell cultures were shown to be responsive to BDNF. Whereas with E17 retinas, the number of surviving Thy-1 positive neurons could be kept constant for at least 4 d, the survival of postnatal neurons was only transiently increased by BDNF. We conclude that in the retina, BDNF affects only the survival of ganglion cells in vitro by a direct action on these cells. The results are discussed in terms of target-derived neurotrophic support during development.


Towards less confusing terminology in reproductive medicine: a proposal.
The use of the term "infertility" and related terms in reproductive medicine is reviewed. Current terminology is found to be ambiguous, confusing and misleading. We recommend that the fertility investigation report of a couple should consist of statements concerning description, diagnosis and prognosis. The description concerns the duration of non-pregnancy before consulting the clinician. A system for prognostic grading is proposed. The fertility investigation report forms the basis for further action, including the possibility of waiting with treatment in case of almost normal or only slightly reduced fertility. The use of the terms infertility, subfertility and fecundity is not necessary, and it is recommended to avoid them.


Seroepidemiology of enterovirus A71 infection in prospective cohort studies of children in southern China, 2013-2018.
Enterovirus A71 (EV-A71)-related hand, foot, and mouth disease (HFMD) imposes a substantial clinical burden in the Asia Pacific region. To inform policy on the introduction of the EV-A71 vaccine into the National Immunization Programme, we investigated the seroepidemiological characteristics of EV-A71 in two prospective cohorts of children in southern China conducted between 2013 and 2018. Our results show that maternal antibody titres declined rapidly in neonates, with over half becoming susceptible to EV-A71 at 1 month of age. Between 6 months and 2 years of age, over 80% of study participants were susceptible, while one third remained susceptible at 5 years old. The highest incidence of EV-A71 infections was observed in children aged 5-6 months. Our findings support EV-A71 vaccination before 6 months for birth cohorts in southern China, potentially with a one-time catch-up vaccination for children 6 months-5 years old. More regionally representative longitudinal seroepidemiological studies are needed to further validate these findings.


Too much or too little? A review of the conundrum of selenium.
Chemical elements such as selenium, fluoride, iron, calcium and magnesium are essential to the human being, although some are toxic when absorbed in high doses. In this paper, the risks associated with insufficient and excessive intake of selenium in the diet are reviewed, focusing on drinking water. Two different approaches are used to derive recommended nutrient intakes (RNI) for adequate nutritional status and guideline values to prevent excessive exposure. The former is based on the daily intake which meets the nutrient requirements of 97.5% of the population. The latter is a value derivation based on an assumed daily per capita consumption at the individual level, a conservative approach used where there is any uncertainty and is related to a negligible risk to health at population level across life stages. There is an increasing need to develop a conceptual framework bringing together aspects of toxicity and essentiality especially for elements apparently exhibiting narrow or overlapping ranges between essentiality and toxicity and to provide guidance on the nature and severity of risks in order to better protect human. While there are a number of frameworks available, these generally only consider food. There is a need to include water, which can be a significant source in some circumstances.


Facing COVID-19 in Ophthalmology Department.
Purpose: To provide useful guidelines, targeted at ophthalmology professionals, to minimize COVID-19 infection of both health-care workers and patients.Methods: In this review we present updated literature merged with our experience from hospitals in Bergamo, the epicenter of the COVID-19 European outbreak.Results: Non-pharmaceutical interventions, hygienic recommendations and personal protective equipment to contain viral spread as well as a suggested risk assessment for postponement of non-urgent cases should be applied in ophthalmologist activity. A triage for ophthalmic outpatient clinic is mandatory.Conclusion: Ophthalmology practice should be reorganized in order to face COVID-19.


Incidence and Causes of Postharvest Fruit Rot in Stored Michigan Cranberries.
The incidence of postharvest fruit rot and associated fungi was studied in stored cranberries in Michigan in 2000 and 2001. Ripe cranberries were harvested from eight commercial farms in southwest and northeast Michigan, including the Upper Peninsula. Eight cranberry cultivars were represented: Stevens, Searles, Le Munyon, Pilgrim, Ben Lear, Bergman, Beckwith, and WSU 61. Fruit rot incidence was assessed within 1 week after harvest. Remaining sound fruit was stored for 2 months at 5°C, and fungi were isolated from rotted fruit after 1 and 2 months of storage. Year and region, but not cultivar, significantly affected the overall rate of rot development in storage. Storage rot levels generally were lower in 2001 than in 2000, particularly in southern Michigan. A high incidence of field rot at harvest did not necessarily lead to a high incidence of storage rot. Storage rot tended to be more severe in the northern than in the southern growing region. Fungi most frequently associated with storage rot were Fusicoccum putrefaciens, Colletotrichum acutatum, Coleophoma empetri, Phomopsis vaccinii, and Phyllosticta elongata. F. putrefaciens was the predominant storage rot fungus in northern Michigan in both years and caused up to 80% fruit rot in storage. C. empetri and P. elongata also were isolated more frequently from beds in northern than southern Michigan in 2001. The cvs. Pilgrim and Stevens were more susceptible to storage rot caused by Colletotrichum acutatum, and Bergman and WSU 61 were more susceptible to storage rot caused by Phomopsis vaccinii than some of the other cultivars.


An anatomically oriented breast coordinate system for mammogram analysis.
We have developed a breast coordinate system that is based on breast anatomy to register female breasts into a common coordinate frame in 2-D mediolateral (ML) or mediolateral oblique (MLO) view mammograms. The breasts are registered according to the location of the pectoral muscle and the nipple and the shape of the breast boundary because these are the most robust features independent of the breast size and shape. On the basis of these landmarks, we have constructed a nonlinear mapping between the parameter frame and the breast region in the mammogram. This mapping makes it possible to identify the corresponding positions and orientations among all of the ML or MLO mammograms, which facilitates an implicit use of the registration, i.e., no explicit image warping is needed. We additionally show how the coordinate transform can be used to extract Gaussian derivative features so that the feature positions and orientations are registered and extracted without nonlinearly deforming the images. We use the proposed breast coordinate transform in a cross-sectional breast cancer risk assessment study of 490 women, in which we attempt to learn breast cancer risk factors from mammograms that were taken prior to when the breast cancer became visible to a radiologist. The coordinate system provides both the relative position and orientation information on the breast region from which the features are derived. In addition, the coordinate system can be used in temporal studies to pinpoint anatomically equivalent locations between the mammograms of each woman and among the mammograms of all of the women in the study. The results of the cross-sectional study show that the classification into cancer and control groups can be improved by using the new coordinate system, compared to other systems evaluated. Comparisons were performed using the area-under-the-receiver-operating-characteristic-curve score. In general, the new coordinate system makes an accurate anatomical registration of breasts possible, which suggests its wide applicability wherever 2-D mammogram registration is required.


Supplementation with the omega-3 long chain polyunsaturated fatty acids: Changes in the concentrations of omega-3 index, fatty acids and molecular phospholipids of people at ultra high risk of developing psychosis.
Omega-3 long chain polyunsaturated fatty acids (n-3 LCPUFA) are necessary for optimum mental health, with recent studies showing low n-3 LCPUFA in people at ultra-high risk (UHR) of developing psychosis. Furthermore, people at UHR of psychosis had increased erythrocyte sphingomyelin (SM) and reduced phosphatidylethanolamine (PE) concentrations as well as 27 erythrocyte phospholipid species that differed when compared to erythrocytes from age matched people not at UHR of psychosis. The aim of this analysis was to evaluate the effect of n-3 supplementation on the different erythrocyte lipid species (including SM and PE concentrations) in people at UHR of psychosis. Participants were randomly assigned to fish oil (containing 840 mg EPA and 560 mg DHA per day) or placebo (paraffin oil) for 6 months. Fasted blood samples were taken at baseline and post intervention. Mass spectrometry was used to analyse the molecular phospholipids and fatty acid composition of erythrocytes for both groups. The n-3 index was significantly increased from 3.0% to 4.12% after 6 months of receiving n-3 capsules. Fish oil capsules increased the phospholipid molecular species containing n-3 LCPUFA, and concomitant decreases in n-6 LCPUFA species. SM species did not show any significant changes in n-3 LCPUFA group however, three SM species (SM 16:0, SM 18:0, SM 18:1) significantly increased after 6 months of supplementation with placebo. N-3 supplementation for 6 months led to higher n-3 incorporation into erythrocytes, at the expense of n-6 PUFA across all phospholipid classes analyzed and may have prevented the increase in SM seen in the placebo group.


A highly tilted membrane configuration for the prefusion state of synaptobrevin.
The SNARE complex plays a vital role in vesicle fusion arising during neuronal exocytosis. Key components in the regulation of SNARE complex formation, and ultimately fusion, are the transmembrane and linker regions of the vesicle-associated protein, synaptobrevin. However, the membrane-embedded structure of synaptobrevin in its prefusion state, which determines its interaction with other SNARE proteins during fusion, is largely unknown. This study reports all-atom molecular-dynamics simulations of the prefusion configuration of synaptobrevin in a lipid bilayer, aimed at characterizing the insertion depth and the orientation of the protein in the membrane, as well as the nature of the amino acids involved in determining these properties. By characterizing the structural properties of both wild-type and mutant synaptobrevin, the effects of C-terminal additions on tilt and insertion depth of membrane-embedded synaptobrevin are determined. The simulations suggest a robust, highly tilted state for membrane-embedded synaptobrevin with a helical connection between the transmembrane and linker regions, leading to an apparently new characterization of structural elements in prefusion synaptobrevin and providing a framework for interpreting past mutation experiments.


Utility of staging laparoscopy in subsets of peripancreatic and biliary malignancies.
To determine the relative benefit of staging laparoscopy in peripancreatic and biliary malignancies. Staging laparoscopy has been used in a variety of peripancreatic and biliary malignancies. The utility of the technique in subsets of these types of cancer has not been systematically compared. One hundred fifty-seven patients underwent laparoscopy after conventional tumor staging; 89 were also staged with laparoscopic ultrasonography. Diagnostic categories were cancer of the pancreatic head and uncinate process, cancer of the body and tail of pancreas, cancer of the extrahepatic bile duct, cancer of the gallbladder, and cancer of the ampulla of Vater/duodenum. In patients with cancer of the head of the pancreas, metastatic disease or vascular invasion was discovered frequently by laparoscopy (31%), whereas in ampullary/duodenal cancer it was never found. The laparoscopic findings in cancer of the head of the pancreas had an important influence on treatment decisions, whereas in cancer of the ampulla/duodenum, laparoscopy had no effect on clinical decisions. Laparoscopy also substantially influenced the treatment of gallbladder cancer; in other tumor types, results were intermediate. Laparoscopic ultrasonography was valuable in cancer of the head of the pancreas. The utility of staging laparoscopy depends on diagnosis. It is recommended for continued use in pancreatic head and gallbladder cancers but not in ampullary malignancies.


Childhood peer status and the clustering of social, economic, and health-related circumstances in adulthood.
Within the school-class context, children attain a social position in the peer hierarchy to which varying amounts of status are attached. Studies have shown that peer status - i.e. the degree of acceptance and likeability among classmates - is associated with adult health. However, these studies have generally paid little attention to the fact that health problems are likely to coincide with other adverse circumstances within the individual. The overarching aim of the current study was therefore to examine the impact of childhood peer status on the clustering of social, economic, and health-related circumstances in adulthood. Using a 1953 cohort born in Stockholm, Sweden (n = 14,294), four outcome profiles in adulthood were identified by means of latent class analysis: 'Average', 'Low education', 'Unemployment', and 'Social assistance recipiency and mental health problems'. Multinomial regression analysis demonstrated that those with lower peer status had exceedingly higher risks of later ending up in the more adverse clusters. This association remained after adjusting for a variety of family-related and individual factors. We conclude that peer status constitutes a central aspect of children's upbringing with important consequences for life chances.


Focused ultrasound for treatment of bone tumours.
Focused ultrasound (FUS) is a modality with rapidly expanding applications across the field of medicine. Treatment of bone lesions with FUS including both benign and malignant tumours has been an active area of investigation. Recently, as a result of a successful phase III trial, magnetic resonance-guided FUS is now a standardised option for treatment of painful bone metastases. This report reviews the clinical applications amenable to treatment with FUS and provides background on FUS and image guidance techniques, results of clinical studies, and future directions. A comprehensive literature search and review of abstracts presented at the recently completed fourth International Focused Ultrasound Symposium was performed. Case reports and older publications revisited in more recent studies were excluded. For clinical studies that extend beyond bone tumours, only the data regarding bone tumours are presented. Fifteen studies assessing the use of focused ultrasound in treatment of primary benign bone tumours, primary malignant tumours, and metastatic tumours meeting the search criteria were identified. For these clinical studies the responders group varied within 91-100%, 85-87% and 64-94%, respectively. Major complications were reported in the ranges 0%, 0-28% and 0-4% for primary benign, malignant and metastatic tumours, respectively. Image-guided FUS is both safe and effective in the treatment of primary and secondary tumours. Additional phase III trials are warranted to more fully define the role of FUS in treatment of both benign and malignant bone tumours.


Simulation study on the heating of the surrounding anatomy during transurethral ultrasound prostate therapy: a 3D theoretical analysis of patient safety.
MRI-guided transurethral ultrasound therapy can generate highly accurate volumes of thermal coagulation conforming to 3D human prostate geometries. This work simulated, quantified, and evaluated the thermal impact of these treatments on the rectum, pelvic bone, neurovascular bundles (NVBs), and urinary sphincters because damage to these structures can lead to complications. Twenty 3D anatomical models of prostate cancer patients were used with detailed bioacoustic simulations incorporating an active feedback algorithm which controlled a rotating, planar ultrasound transducer (17, 4 x 3 mm2 elements, 10 W(acoustic)/cm2). Heating of the adjacent surrounding anatomy was evaluated at 4.7, 9.7, and 14.2 MHz using thermal tolerances reported in literature. Heating of the rectum posed the most important safety concern, influenced largely by the water temperature of an endorectal cooling device (ECD); depending on anatomy, temperatures of 7-37 degrees C were required to limit potential damage to less than 10 mm3 on the outer 1 mm layer of the rectal wall. Heating of the pelvic bone could be important at 4.7 MHz. A smaller sized ECD or a higher ultrasound frequency in sectors where the bone was less than 10 mm from the prostate reduced heating in all cases below the threshold for irreversible damage. Heating of the NVB was significant in 75% of the patient models in the absence of treatment planning; this proportion was reduced to 5% by increasing treatment margins up to 4 mm. To avoid damaging the urinary sphincters, the transducer should be positioned at least 2-4 mm from the sphincters, depending on the transurethral cooling temperature. Simulations show that MRI-guided transurethral therapy can treat the prostate accurately, but in the absence of treatment planning, some thermal impact can be predicted on the surrounding anatomy. Treatment planning strategies have been developed, which reduce thermal injury to the surrounding anatomy.


Biotic ligand model of the acute toxicity of metals. 2. Application to acute copper toxicity in freshwater fish and Daphnia.
The biotic ligand model (BLM) was developed to explain and predict the effects of water chemistry on the acute toxicity of metals to aquatic organisms. The biotic ligand is defined as a specific receptor within an organism where metal complexation leads to acute toxicity. The BLM is designed to predict metal interactions at the biotic ligand within the context of aqueous metal speciation and competitive binding of protective cations such as calcium. Toxicity is defined as accumulation of metal at the biotic ligand at or above a critical threshold concentration. This modeling framework provides mechanistic explanations for the observed effects of aqueous ligands, such as natural organic matter, and water hardness on metal toxicity. In this paper, the development of a copper version of the BLM is described. The calibrated model is then used to calculate LC50 (the lethal concentration for 50% of test organisms) and is evaluated by comparison with published toxicity data sets for freshwater fish (fathead minnow, Pimephales promelas) and Daphnia.


Lacking ketohexokinase-A exacerbates renal injury in streptozotocin-induced diabetic mice.
Ketohexokinase (KHK), a primary enzyme in fructose metabolism, has two isoforms, namely, KHK-A and KHK-C. Previously, we reported that renal injury was reduced in streptozotocin-induced diabetic mice which lacked both isoforms. Although both isoforms express in kidney, it has not been elucidated whether each isoform plays distinct roles in the development of diabetic kidney disease (DKD). The aim of the study is to elucidate the role of KHK-A for DKD progression. Diabetes was induced by five consecutive daily intraperitoneal injections of streptozotocin (50 mg/kg) in C57BL/6J wild-type mice, mice lacking KHK-A alone (KHK-A KO), and mice lacking both KHK-A and KHK-C (KHK-A/C KO). At 35 weeks, renal injury, inflammation, hypoxia, and oxidative stress were examined. Metabolomic analysis including polyol pathway, fructose metabolism, glycolysis, TCA (tricarboxylic acid) cycle, and NAD (nicotinamide adenine dinucleotide) metabolism in kidney and urine was done. Diabetic KHK-A KO mice developed severe renal injury compared to diabetic wild-type mice, and this was associated with further increases of intrarenal fructose, dihydroxyacetone phosphate (DHAP), TCA cycle intermediate levels, and severe inflammation. In contrast, renal injury was prevented in diabetic KHK-A/C KO mice compared to both wild-type and KHK-A KO diabetic mice. Further, diabetic KHK-A KO mice contained decreased renal NAD+ level with the increase of renal hypoxia-inducible factor 1-alpha expression despite having increased renal nicotinamide (NAM) level. These results suggest that KHK-C might play a deleterious role in DKD progression through endogenous fructose metabolism, and that KHK-A plays a unique protective role against the development of DKD.


Purification and characterization of a sulfated glycoprotein secreted by Sertoli cells.
Sulfated glycoprotein 2 (SGP-2), the major secretion product of Sertoli cells, was purified from cell culture medium by reverse-phase high-performance liquid chromatography. The native protein consists of disulfide-linked monomers of 41,000 and 29,000 daltons which have a strong tendency to associate into multimers. The purified SGP-2 was subjected to amino acid analysis and contained high levels of Asx (11.1%), Glx (15.1%), and leucine (11.5%). The oligosaccharides on the purified SGP-2 were analyzed to determine the monosaccharide compositions and the molecular weights of the intact carbohydrate moieties. SGP-2 was shown to be 23.7% carbohydrate and consisted of 1% fucose, 3.5% mannose, 4.1% galactose, 7.1% N-acetylglucosamine, and 8.0% N-acetylneuraminic acid. No N-acetylgalactosamine was detected. When the SGP-2 was digested with proteases, the intact oligosaccharides were chromatographed over a Bio-Gel P-6 column and found to elute in a single symmetrical peak of approximately 3,300 g/mol. On the basis of these results, the oligosaccharides on SGP-2 were proposed to consist of triantennary chains similar to those found on fetuin. When the 35SO4(2-)-labeled SGP-2 was digested with Pronase, the free amino acids could be separated by chromatography from the oligosaccharide. The 35SO4(2-) was shown to be associated with the oligosaccharide portion of SGP-2.


Recombinant human IgA1 and IgA2 autoantibodies to type VII collagen induce subepidermal blistering ex vivo.
Subepidermal autoimmune blistering dermatoses (AIBD) are prototypic autoantibody-mediated diseases. In epidermolysis bullosa acquisita (EBA), an autoimmune disease with severe and chronic skin blistering, autoantibodies are directed against type VII collagen. IgG is the predominant autoantibody isotype of EBA, the pathogenicity of which has been demonstrated in a variety of in vivo and ex vivo disease models. In contrast, there is not much evidence for the pathogenicity of IgA, which may appear as the only autoantibody isotype in some EBA patients. To investigate the pathogenic potential of IgA autoantibodies, we generated chimeric V gene-matched human IgA1, IgA2, and control IgG1 autoantibodies directed against type VII collagen. Immobilized immune complexes containing the rIgA1 and rIgA2 autoantibodies induced the dose-dependent release of reactive oxygen species from neutrophil granulocytes, a precondition for blister formation. Moreover, both rIgA1 and rIgA2 induced leukocyte-dependent dermal-epidermal separation in cryosections of human skin. In contrast with rIgG1, neither rIgA1 nor rIgA2 was capable of inducing complement deposition at the dermal-epidermal junction. Because complement activation is a prerequisite for blister induction, this lack of function compared with IgG1 may be compensated for by the stronger activation of neutrophil granulocytes by both IgA1 and IgA2. For IgG-mediated AIBD, immunoadsorption therapy is a convenient treatment modality for the removal of pathogenic autoantibodies, particularly in treatment-resistant cases. The results of this study show the pathogenic potential of IgA autoantibodies and support the development of adsorber matrices for IgA-mediated AIBD.


Good news-bad news: the Yin and Yang of immune privilege in the eye.
The eye and the brain are prototypical tissues manifesting immune privilege (IP) in which immune responses to foreign antigens, particularly alloantigens are suppressed, and even completely inhibited. Explanations for this phenomenon are numerous and mostly reflect our evolving understanding of the molecular and cellular processes underpinning immunological responses generally. IP is now viewed as a property of many tissues and the level of expression of IP varies not only with the tissue but with the nature of the foreign antigen and changes in the limited conditions under which privilege can operate as a mechanism of immunological tolerance. As a result, IP functions normally as a homeostatic mechanism preserving normal function in tissues, particularly those with highly specialized function and limited capacity for renewal such as the eye and brain. However, IP is relatively easily bypassed in the face of a sufficiently strong immunological response, and the privileged tissues may be at greater risk of collateral damage because its natural defenses are more easily breached than in a fully immunocompetent tissue which rapidly rejects foreign antigen and restores integrity. This two-edged sword cuts its swathe through the eye: under most circumstances, IP mechanisms such as blood-ocular barriers, intraocular immune modulators, induction of T regulatory cells, lack of lymphatics, and other properties maintain tissue integrity; however, when these are breached, various degrees of tissue damage occur from severe tissue destruction in retinal viral infections and other forms of uveoretinal inflammation, to less severe inflammatory responses in conditions such as macular degeneration. Conversely, ocular IP and tumor-related IP can combine to permit extensive tumor growth and increased risk of metastasis thus threatening the survival of the host.


The contribution of the gut microbiome to neurodevelopment and neuropsychiatric disorders.
Bidirectional communication between the gut and brain is well recognized, with data now accruing for a specific role of the gut microbiota in that link, referred to as the microbiome-gut-brain axis. This review will discuss the emerging role of the gut microbiota in brain development and behavior. Animal studies have clearly demonstrated effects of the gut microbiota on gene expression and neurochemical metabolism impacting behavior and performance. Based on these changes, a modulating role of the gut microbiota has been demonstrated for a variety of neuropsychiatric disorders, including depression, anxiety, and movement including Parkinson's, and importantly for the pediatric population autism. Critical developmental windows that influence early behavioral outcomes have been identified that include both the prenatal environment and early postnatal colonization periods. The clearest data regarding the role of the gut microbiota on neurodevelopment and psychiatric disorders is from animal studies; however, human data have begun to emerge, including an association between early colonization patterns and cognition. The importance of understanding the contribution of the gut microbiota to the development and functioning of the nervous system lies in the potential to intervene using novel microbial-based approaches to treating neurologic conditions. While pathways of communication between the gut and brain are well established, the gut microbiome is a new component of this axis. The way in which organisms that live in the gut influence the central nervous system (CNS) and host behavior is likely to be multifactorial in origin. This includes immunologic, endocrine, and metabolic mechanisms, all of which are pathways used for other microbial-host interactions. Germ-free (GF) mice are an important model system for understanding the impact of gut microbes on development and function of the nervous system. Alternative animal model systems have further clarified the role of the gut microbiota, including antibiotic treatment, fecal transplantation, and selective gut colonization with specific microbial organisms. Recently, researchers have started to examine the human host as well. This review will examine the components of the CNS potentially influenced by the gut microbiota, and the mechanisms mediating these effects. Links between gut microbial colonization patterns and host behavior relevant to a pediatric population will be examined, highlighting important developmental windows in utero or early in development.


Endbulbs of held and spherical bushy cells in cats: morphological correlates with physiological properties.
Single auditory nerve fibers of type I spiral ganglion cells in cats were electrophysiologically characterized by recording with micropipettes inserted into the axon and then labeled by intracellular injections of horseradish peroxidase (HRP) through the same pipettes. This method for staining and studying single neurons allowed us to describe structure-function relationships for labeled endbulbs of Held and the somata of their postsynaptic spherical bushy cells. The silhouette areas of terminal endbulbs and the corresponding somata of spherical bushy cells were determined by planimetry from drawings made with a light microscope and drawing tube. On the presynaptic side, endbulb area is related to fiber characteristic frequency (CF, the frequency to which a fiber is most sensitive) such that the largest endbulbs arise from fibers having CFs between 1 and 4 kHz; smaller endbulbs can arise from fibers of any CF. Endbulb area is not correlated with fiber spontaneous discharge rate (SR). Dividing the endbulb's silhouette area by its silhouette perimeter, however, yields a "form factor" that is a reliable indicator of fiber SR: Endbulbs from fibers of low-medium SR (less than or equal to 18 spikes/second) have form factor values less than 0.52, whereas endbulbs of high SR fibers (greater than 18 spikes/second) have values greater than 0.52. This form factor should therefore be predictive of SR groupings in auditory fibers for which physiological data are not available. On the postsynaptic side, the somata of spherical bushy cells receiving endbulbs from low-medium SR fibers are on average smaller than those receiving endbulbs from high SR fibers. In contrast, the nuclei of the spherical bushy cells are the same size regardless of presynaptic fiber SR. Some of the effects of low-medium SR fibers on their postsynaptic targets, when compared to those of high SR fibers, appear to be mimicked by effects of experimentally induced deprivation.


An explanatory model of health promotion and quality of life for persons with post-polio syndrome.
Although previous studies have examined selected factors influencing health promoting behaviors or quality of life, the complex interplay of these variables in persons with different chronic disabling conditions has not been investigated. This study tested an explanatory model of variables influencing health promotion and quality of life originally developed with a sample of persons with multiple sclerosis in a new sample of persons living with post-polio syndrome (PPS) in the USA. The sample of 1603 persons with PPS (1123 females, 478 males and 2 unknown) completed a battery of instruments including measures of severity of polio-related impairment, barriers to health promoting behaviors, resources, self-efficacy, acceptance, health promoting behaviors and perceived quality of life. A model originally developed in a sample of 786 persons with multiple sclerosis was assessed and modified using the weighted least squares procedure (WLS) which is implemented by LISREL8. The structural equation analyses resulted in a proper solution that exhibited adequate fit: chi2 (8, N = 1549)=84.22, p<0.05; GFI=0.96, IFI=0.90, CFI=0.90. The antecedent variables accounted for 65% of the variance in the frequency of health promoting behaviors and 53% of the variance in perceived quality of life. The model test supports the hypothesis that quality of life is the outcome of a complex interplay between contextual factors (severity of impairment), antecedent variables, and health promoting behaviors. It also suggests that the relationships among these variables are similar in samples of persons with two different chronic conditions. Further research using a qualitative approach is needed to clarify other contributors to quality of life in persons with post-polio syndrome.


Intrinsic apoptosis in mechanically ventilated human diaphragm: linkage to a novel Fos/FoxO1/Stat3-Bim axis.
Mechanical ventilation (MV) is a life-saving measure in many critically ill patients. However, prolonged MV results in diaphragm dysfunction that contributes to the frequent difficulty in weaning patients from the ventilator. The molecular mechanisms underlying ventilator-induced diaphragm dysfunction (VIDD) remain poorly understood. We report here that MV induces myonuclear DNA fragmentation (3-fold increase; P<0.01) and selective activation of caspase 9 (P<0.05) and Bcl2-interacting mediator of cell death (Bim; 2- to 7-fold increase; P<0.05) in human diaphragm. MV also statistically significantly down-regulates mitochondrial gene expression and induces oxidative stress. In cultured muscle cells, we show that oxidative stress activates each of the catabolic pathways thought to underlie VIDD: apoptotic (P<0.05), proteasomal (P<0.05), and autophagic (P<0.01). Further, silencing Bim expression blocks (P<0.05) oxidative stress-induced apoptosis. Overlapping the gene expression profiles of MV human diaphragm and H₂O₂-treated muscle cells, we identify Fos, FoxO1, and Stat3 as regulators of Bim expression as well as of expression of the catabolic markers atrogin and LC3. We thus identify a novel Fos/FoxO1/Stat3-Bim intrinsic apoptotic pathway and establish the centrality of oxidative stress in the development of VIDD. This information may help in the design of specific drugs to prevent this condition.


Swept-3-D Ultrasound Imaging of the Mouse Brain Using a Continuously Moving 1-D-Array-Part II: Functional Imaging.
Functional ultrasound (fUS) using a 1-D-array transducer normally is insufficient to capture volumetric functional activity due to being restricted to imaging a single brain slice at a time. Typically, for volumetric fUS, functional recordings are repeated many times as the transducer is moved to a new location after each recording, resulting in a nonunique average mapping of the brain response and long scan times. Our objective was to perform volumetric 3-D fUS in an efficient and cost-effective manner. This was achieved by mounting a 1-D-array transducer to a high-precision motorized linear stage and continuously translating over the mouse brain in a sweeping manner. We show how the speed at which the 1-D-array is translated over the brain affects the sampling of the hemodynamic response (HR) during visual stimulation as well as the quality of the resulting power Doppler image (PDI). Functional activation maps were compared between stationary recordings, where only one functional slice is obtained for every recording, and our swept-3-D method, where volumetric fUS was achieved in a single functional recording. The results show that the activation maps obtained with our method closely resemble those obtained during a stationary recording for that same location, while our method is not restricted to functional imaging of a single slice. Lastly, a mouse brain subvolume of ~6 mm is scanned at a volume rate of 1.5 s per volume, with a functional PDI reconstructed every [Formula: see text], highlighting swept-3-D's potential for volumetric fUS. Our method provides an affordable alternative to volumetric fUS using 2-D-matrix transducers, with a high SNR due to using a fully sampled 1-D-array transducer, and without the need to repeat functional measurements for every 2-D slice, as is most often the case when using a 1-D-array. This places our swept-3-D method as a potentially valuable addition to conventional 2-D fUS, especially when investigating whole-brain functional connectivity, or when shorter recording durations are desired.


Transcriptional activity, chromosomal distribution and expression effects of transposable elements in Coffea genomes.
Plant genomes are massively invaded by transposable elements (TEs), many of which are located near host genes and can thus impact gene expression. In flowering plants, TE expression can be activated (de-repressed) under certain stressful conditions, both biotic and abiotic, as well as by genome stress caused by hybridization. In this study, we examined the effects of these stress agents on TE expression in two diploid species of coffee, Coffea canephora and C. eugenioides, and their allotetraploid hybrid C. arabica. We also explored the relationship of TE repression mechanisms to host gene regulation via the effects of exonized TE sequences. Similar to what has been seen for other plants, overall TE expression levels are low in Coffea plant cultivars, consistent with the existence of effective TE repression mechanisms. TE expression patterns are highly dynamic across the species and conditions assayed here are unrelated to their classification at the level of TE class or family. In contrast to previous results, cell culture conditions per se do not lead to the de-repression of TE expression in C. arabica. Results obtained here indicate that differing plant drought stress levels relate strongly to TE repression mechanisms. TEs tend to be expressed at significantly higher levels in non-irrigated samples for the drought tolerant cultivars but in drought sensitive cultivars the opposite pattern was shown with irrigated samples showing significantly higher TE expression. Thus, TE genome repression mechanisms may be finely tuned to the ideal growth and/or regulatory conditions of the specific plant cultivars in which they are active. Analysis of TE expression levels in cell culture conditions underscored the importance of nonsense-mediated mRNA decay (NMD) pathways in the repression of Coffea TEs. These same NMD mechanisms can also regulate plant host gene expression via the repression of genes that bear exonized TE sequences.


Progression of liver disease in children and adults with lysosomal acid lipase deficiency.
Manifestations of the autosomal recessive disorder lysosomal acid lipase deficiency (LAL-D) include hepatomegaly, elevated serum liver enzymes, and progressive liver disease. We report an analysis of time to progression from first clinical manifestation to first documentation of hepatic fibrosis, cirrhosis, or liver transplantation from an observational study of pediatric and adult patients with LAL-D (clinical trial registration: NCT01528917). Data were analyzed from 31 patients with available biopsy data and 1 patient without biopsy data who had undergone liver transplantation. Time to first documentation of fibrosis, cirrhosis, or liver transplantation following the first LAL-D clinical manifestation was estimated using Kaplan-Meier analysis. The median time to an event was 3.1 years. These findings illustrate the progression of liver damage in LAL-D and the elevated risk for liver transplantation among children and adults with LAL-D.


Thrombo-embolism and antithrombotic therapy for heart failure in sinus rhythm. A joint consensus document from the ESC Heart Failure Association and the ESC Working Group on Thrombosis.
Chronic heart failure (HF) with either reduced or preserved ejection fraction is common and remains an extremely serious disorder with a high mortality and morbidity. Many complications related to HF can be related to thrombosis. Epidemiological and pathophysiological data also link HF to an increased risk of thrombosis, leading to the clinical consequences of sudden death, stroke, systemic thrombo-embolism, and/or venous thrombo-embolism. This consensus document of the Heart Failure Association (EHFA) of the European Society of Cardiology (ESC) and the ESC Working Group on Thrombosis reviews the published evidence and summarizes 'best practice', and puts forward consensus statements that may help to define evidence gaps and assist management decisions in everyday clinical practice. In HF patients with atrial fibrillation, oral anticoagulation is recommended, and the CHA(2)DS(2)-VASc and HAS-BLED scores should be used to determine the likely risk-benefit ratio (thrombo-embolism prevention vs. risk of bleeding) of oral anticoagulation. In HF patients with reduced left ventricular ejection fraction who are in sinus rhythm there is no evidence of an overall benefit of vitamin K antagonists (e.g. warfarin) on mortality, with risk of major bleeding. Despite the potential for a reduction in ischaemic stroke, there is currently no compelling reason to use warfarin routinely for these patients. Risk factors associated with increased risk of thrombo-embolic events should be identified and decisions regarding use of anticoagulation individualized. Patient values and preferences are important determinants when balancing the risk of thrombo-embolism against bleeding risk. New oral anticoagulants that offer a different risk-benefit profile compared with warfarin may appear as an attractive therapeutic option, but this would need to be confirmed in clinical trials.


Levels and patterns of DNA variation in two sympatric mangrove species, Rhizophora apiculata and R. mucronata from Thailand.
In mangrove species the past geomorphic changes in coastal regions and reproductive systems are important factors of their distribution and genetic structure of populations. However, very little is known about the levels of genetic variation of Rhiozophora species in Southeast Asia. In this study, we surveyed levels and patterns of genetic variation as well as population structure of two sympatric mangrove species, Rhizophora apiculata and R. mucronata in Thailand, using five nuclear genes and two cpDNA regions. In all investigated DNA regions, nucleotide variation within species was low, while nucleotide divergence between the two species was considerable. The nuclear genes evolved 10 times faster than the cpDNA regions. In both R. apiculata and R. mucronata, significant positive F(IS) values were found, indicating deviation from Hardy-Weinberg proportions and a deficiency of heterozygotes. In both species, we found significant genetic differentiation between populations. However, the pattern of population differentiation (F(ST)) of R. apiculata differed from that of R. mucronata. Our results suggest that the two investigated species have different demographic history, even though they are sympatric and have similar reproductive systems.


Metabolomics reveals a role for the chromatin-binding protein HMGN5 in glutathione metabolism.
High mobility group nucleosome-binding protein 5 (HMGN5) is a chromatin architectural protein that binds specifically to nucleosomes and reduces the compaction of the chromatin fiber. The protein is present in most vertebrate tissues however the physiological function of this protein is unknown. To examine the function of HMGN5 in vivo, mice lacking the nucleosome-binding domain of HMGN5 were generated and characterized. Serological analysis revealed that compared to wild-type littermates (Hmgn5(+/Y)), mice with a targeted mutation in the HMGN5 gene (Hmgn5(tm1/Y)), had elevated serum albumin, non-HDL cholesterol, triglycerides, and alanine transaminase, suggesting mild hepatic abnormalities. Metabolomics analysis of liver extracts and urine revealed clear differences in metabolites between Hmgn5(tm1/Y) and their Hmgn5(+/Y) littermates. Hmgn5(tm1/Y) mice had a significant increase in hepatic glutathione levels and decreased urinary concentrations of betaine, phenylacetylglycine, and creatine, all of which are metabolically related to the glutathione precursor glycine. Microarray and qPCR analysis revealed that expression of two genes affecting glutathione metabolism, glutathione peroxidase 6 (Gpx6) and hexokinase 1 (Hk1), was significantly decreased in Hmgn5(tm1/Y) mouse liver tissue. Analysis of chromatin structure by DNase I digestion revealed alterations in the chromatin structure of these genes in the livers of Hmgn5(tm1/Y) mice. Thus, functional loss of HMGN5 leads to changes in transcription of Gpx6 and Hk1 that alter glutathione metabolism.


Vulval elephantiasis as a result of tubercular lymphadenitis: two case reports and a review of the literature.
Elephantiasis as a result of chronic lymphedema is characterized by gross enlargement of the arms, legs or genitalia, and occurs due to a variety of obstructive diseases of the lymphatic system. Genital elephantiasis usually follows common filariasis and lymphogranuloma venereum. It may follow granuloma inguinale, carcinomas, lymph node dissection or irradiation and tuberculosis but this happens rarely. Vulval elephantiasis as a consequence of extensive lymph node destruction by tuberculosis is very rare. We present two very unusual cases of vulval elephantiasis due to tuberculous destruction of the inguinal lymph nodes. Two Indian women - one aged 40 years and the other aged 27 years, with progressively increasing vulval swellings over a period of five and four years respectively - presented to our hospital. In both cases, there was a significant history on presentation. Both women had previously taken a complete course of anti-tubercular treatment for generalized lymphadenopathy. The vulval swellings were extremely large: in the first case report, measuring 35 × 25 cm on the right side and 45 × 30 cm on the left side, weighing 20 lb and 16 lb respectively. Both cases were managed by surgical excision with reconstruction and the outcome was positive. Satisfactory results have been maintained during a follow-up period of six years in both cases. Elephantiasis of the female genitalia is unusual and it has rarely been reported following tuberculosis. We report two cases of vulval elephantiasis as a consequence of extensive lymph node destruction by tuberculosis, in order to highlight this very rare clinical scenario.


CACNA1A mutations causing episodic and progressive ataxia alter channel trafficking and kinetics.
CACNA1A encodes CaV2.1, the pore-forming subunit of P/Q-type voltage-gated calcium channel complexes. Mutations in CACNA1A cause a wide range of neurologic disturbances variably associated with cerebellar degeneration. Functional studies to date focus on electrophysiologic defects that do not adequately explain the phenotypic findings. To investigate whether some missense mutations might interfere with protein folding and trafficking, eventually leading to protein aggregation and neuronal injury. The authors studied the functional consequences of two pore missense mutations, C287Y and G293R, in two families with EA2, one newly discovered and the other previously reported. Both mutations caused episodic and interictal ataxia. The biophysical properties of mutant and wild type calcium channels were examined by whole-cell patch-clamp recordings in transfected COS-7 cells. The plasma membrane targeting was visualized by confocal fluorescence imaging on CaV2.1 tagged with green fluorescent protein. The mutant channels exhibited a marked reduction in current expression and deficiencies in plasma membrane targeting. In addition to altered channel function, the deficiency in protein misfolding and trafficking associated with the C287Y and G293R mutants may contribute to the slowly progressive cerebellar ataxia.


Targeting rubella for elimination.
Rubella is an acute, usually mild viral disease. However, when rubella infection occurs just before conception or during the first 8-10 weeks of gestation, it causes multiple fetal defects in up to 90% of cases, known as Congenital Rubella Syndrome (CRS). It may result in fetal wastage, stillbirths and sensorineural hearing deficit up to 20 weeks of gestation. Rubella vaccine (RA 27/3) is highly effective and has resulted in elimination of rubella and CRS from the western hemisphere and several European countries. Review of several studies documents the duration of protection over 10-21 years following one dose of RA27/3 vaccination, and persistent seropositivity in over 95% cases. Studies in India show seronegativity to rubella among adolescent girls to vary from 10% to 36%. Although due to early age of infection resulting in protection in the reproductive age group, incidence of rubella in India is not very high. However, due to severity of CRS coupled with introduction of RCV in private sector and in some of the states which is likely to lead to sub-optimal coverage and resulting higher risk of rubella during pregnancy in the coming decades, it is imperative to adopt the goal of rubella elimination. As in order to control measles, the country has adopted strategy of delivering second dose of measles through measles campaigns covering children 9 months to 10 years of age in 14 states, it is recommended to synergize efforts for elimination of rubella with these campaigns by replacing measles vaccine by MR or MMR vaccine. Other states which are to give second dose of measles through routine immunization will also have to adopt campaign mode in order to eliminate rubella from the country over 10-20 years. Subsequently, measles vaccine can be replaced by MR or MMR vaccine in the national schedule.


Israeli oncology and nononcology nurses' attitudes toward physician-assisted dying: a comparison study.
To compare the attitudes of Israeli oncology and nononcology nurses toward physician-assisted dying (PAD) and its legalization and to determine the factors that may be related to their attitudes. Nonrandomized, correlational study. A teaching hospital in Jerusalem, Israel. 71 oncology nurses and 52 nurses working in the maternity and nursery departments. The majority of nurses supported PAD and its legalization, with the greatest support being given when a suffering patient was involved. In the vignettes dealing with the nurse-patient relationship, oncology nurses were significantly less likely than nononcology nurses to agree with PAD but more likely to stay with their patients while the lethal drug was being given. Nononcology nurses were more supportive of legalization than oncology nurses. Religious nurses and nurses who observed religious traditions were significantly less likely to support PAD or vote for its legalization. Israeli nurses generally are supportive of PAD. However, oncology nurses were less likely to support its practice. In addition, the more religious nurses considered themselves, the less likely they were to support the practice. Israeli nurses need to be made aware of the ethical, social, and legal implications of PAD and its legalization for the Israeli nursing profession. In addition, more cross-cultural research on attitudes toward PAD needs to be conducted.