Inhibition of Wnt signaling by ICAT, a novel beta-catenin-interacting protein.
Wnt signaling has an important role in both embryonic development and tumorigenesis. beta-Catenin, a key component of the Wnt signaling pathway, interacts with the TCF/LEF family of transcription factors and activates transcription of Wnt target genes. Here, we identify a novel beta-catenin-interacting protein, ICAT, that was found to inhibit the interaction of beta-catenin with TCF-4 and represses beta-catenin-TCF-4-mediated transactivation. Furthermore, ICAT inhibited Xenopus axis formation by interfering with Wnt signaling. These results suggest that ICAT negatively regulates Wnt signaling via inhibition of the interaction between beta-catenin and TCF and is integral in development and cell proliferation.


Genetic Diversity of the Amur Tiger (Panthera tigris altaica): Are There Differences between the 20th and the Early 21st Centuries?
Polymorphism of nine microsatellite loci in the Sikhote-Alin tiger population was analyzed in the modern and recent historical periods, using blood, excrement, and museum bone samples. Tests for excess heterozygosity to determine whether the population went through a period of low abundance and a low value of the Garza-Williamson coefficient indicated that such events were highly probable to occur in both recent and earlier history. The mean effective population size Ne of a current sample was 34.4 (95% confidence interval 17-106.8). This fact is of great interest in the contest of conservation and restoration of endangered large cat species.


Histone H3K56 acetylation, Rad52, and non-DNA repair factors control double-strand break repair choice with the sister chromatid.
DNA double-strand breaks (DSBs) are harmful lesions that arise mainly during replication. The choice of the sister chromatid as the preferential repair template is critical for genome integrity, but the mechanisms that guarantee this choice are unknown. Here we identify new genes with a specific role in assuring the sister chromatid as the preferred repair template. Physical analyses of sister chromatid recombination (SCR) in 28 selected mutants that increase Rad52 foci and inter-homolog recombination uncovered 8 new genes required for SCR. These include the SUMO/Ub-SUMO protease Wss1, the stress-response proteins Bud27 and Pdr10, the ADA histone acetyl-transferase complex proteins Ahc1 and Ada2, as well as the Hst3 and Hst4 histone deacetylase and the Rtt109 histone acetyl-transferase genes, whose target is histone H3 Lysine 56 (H3K56). Importantly, we use mutations in H3K56 residue to A, R, and Q to reveal that H3K56 acetylation/deacetylation is critical to promote SCR as the major repair mechanism for replication-born DSBs. The same phenotype is observed for a particular class of rad52 alleles, represented by rad52-C180A, with a DSB repair defect but a spontaneous hyper-recombination phenotype. We propose that specific Rad52 residues, as well as the histone H3 acetylation/deacetylation state of chromatin and other specific factors, play an important role in identifying the sister as the choice template for the repair of replication-born DSBs. Our work demonstrates the existence of specific functions to guarantee SCR as the main repair event for replication-born DSBs that can occur by two pathways, one Rad51-dependent and the other Pol32-dependent. A dysfunction can lead to genome instability as manifested by high levels of homolog recombination and DSB accumulation.


Plasticity of binocularity and visual acuity are differentially limited by nogo receptor.
The closure of developmental critical periods consolidates neural circuitry but also limits recovery from early abnormal sensory experience. Degrading vision by one eye throughout a critical period both perturbs ocular dominance (OD) in primary visual cortex and impairs visual acuity permanently. Yet understanding how binocularity and visual acuity interrelate has proven elusive. Here we demonstrate the plasticity of binocularity and acuity are separable and differentially regulated by the neuronal nogo receptor 1 (NgR1). Mice lacking NgR1 display developmental OD plasticity as adults and their visual acuity spontaneously improves after prolonged monocular deprivation. Restricting deletion of NgR1 to either cortical interneurons or a subclass of parvalbumin (PV)-positive interneurons alters intralaminar synaptic connectivity in visual cortex and prevents closure of the critical period for OD plasticity. However, loss of NgR1 in PV neurons does not rescue deficits in acuity induced by chronic visual deprivation. Thus, NgR1 functions with PV interneurons to limit plasticity of binocularity, but its expression is required more extensively within brain circuitry to limit improvement of visual acuity following chronic deprivation.


"Support Your Client at the Space That They're in": HIV Pre-Exposure Prophylaxis (PrEP) Prescribers' Perspectives on PrEP-Related Risk Compensation.
Despite the demonstrated effectiveness of HIV pre-exposure prophylaxis (PrEP) and evidence that most PrEP users do not engage in risk compensation (i.e., increased risk behavior due to a perceived decrease in HIV susceptibility), some healthcare providers report patient risk compensation to be a deterrent to prescribing PrEP. Overcoming this barrier is essential to supporting PrEP access and uptake among people at risk for HIV. To inform such efforts, this qualitative study explored PrEP-related risk compensation attitudes among providers with firsthand experience prescribing PrEP. US-based PrEP providers (n = 18), most of whom were HIV specialists, were recruited through direct outreach and referral from colleagues and other participants. Individual 90-min semistructured interviews were conducted by phone or in person from September 2014 through February 2015, transcribed, and thematically analyzed. Three attitudinal themes emerged: (1) providers' role is to support patients in making informed decisions, (2) risk behavior while taking PrEP does not fully offset PrEP's protective benefit (i.e., PrEP confers net protection, even with added behavioral risk), and (3) PrEP-related risk compensation is unduly stigmatized within and beyond the healthcare community. Participants were critical of other healthcare providers' negative judgment of patients and reluctance to prescribe PrEP due to anticipated risk compensation. Several providers also acknowledged an evolution in their thinking from initial ambivalence toward greater acceptance of PrEP and PrEP-related behavior change. PrEP providers' insights about risk compensation may help to address unsubstantiated concerns about PrEP-related risk compensation and challenge the acceptability of withholding PrEP on these grounds.


[Use of near-infrared spectroscopy for control of limb perfusion during venoarterial ECMO treatment : Application and limitations].
Patients undergoing peripheral venoarterial extracorporeal membrane oxygenation have a high risk of lower limb ischemia. In general, regular controls are carried out based on clinical and laboratory parameters in order to quickly detect and treat complications. These controls are challenging due to states of shock, nonpulsatile flow and vasopressor therapy. As additional monitoring the use of near-infrared spectroscopy (NIRS) is described in the literature as being very successful in detecting ischemia. The present article describes the use and possible limitations of NIRS for the diagnostics of peripheral ischemia.


The impact of the Covid-19 pandemic and government intervention on active mobility.
With data from automated counting stations and controlling for weather and calendar effects, we estimate the isolated impacts of the "first wave" of Covid-19 pandemic and subsequent government intervention (contact restrictions and closures of public spaces) on walking and cycling in 10 German cities. Pedestrian traffic in pedestrian zones decreases with higher local incidence values, and with stricter government intervention. There are ambiguous effects for cycling, which decreases in cities with a higher modal share of cycling, and increases in others. Moreover, we find impact heterogeneity with respect to different weekdays and hours of the day, both for cycling and walking. Additionally, we use data on overall mobility changes, which were derived from mobile phone data, in order to estimate the modal share changes of cycling. In almost all cities, the modal share of cycling increases during the pandemic, with higher increases in non-bicycle cities and during stronger lockdown interventions.


Antagonic effect of the inhibition of inducible nitric oxide on the mortality of mice acutely infected with Escherichia coli and Bacteroides fragilis.
Sepsis, the leading cause of death in intensive care units, is associated with overproduction of nitric oxide (NO) due to inducible NO synthase (iNOS), responsible for some of the pathologic changes. Aminoguanidine (AG) is a selective iNOS inhibitor with reported inconsistent actions in sepsis. To investigate the influence of iNOS, we studied models of acute bacterial sepsis using acute challenges with aerobic (Escherichia coli) and anaerobic (Bacteroides fragilis) bacteria in the presence of AG. Six-week-old, 23 g, male and female BALB/c and C57Bl/6j mice, in equal proportions, were inoculated (ip) with bacteria in groups of 4 animals for each dose and each experiment in the absence or presence of AG (50 mg/kg, ip, starting 24 h before challenge and daily until day 6) and serum nitrate was measured by chemiluminescence. Both types of bacteria were lethal to mice, with an LD50 of 6 nephelometric units (U) for E. coli and 8 U for B. fragilis. Nitrate production peaked on the second day after E. coli inoculation with 8 and 6 U (P < 0.05), but was absent after non-lethal lower doses. After challenge with B. fragilis this early peak occurred at all tested doses after 24 h, including non-lethal ones (P < 0.05). AG-treated mice challenged with E. coli presented higher survival (P < 0.05) and increased LD50. AG-treated mice challenged with B. fragilis had lower LD50 and higher mortality. Control AG-treated animals presented no toxic effects. The opposite effect of iNOS blockade by AG in these models could be explained by restriction of oxygen for immune cells or an efficient action of NO in anaerobic localized infections. The antagonic role of NO production observed in our bacterial models could explain the reported discrepancy of NO action in sepsis.


The polymeric immunoglobulin receptor (secretory component) mediates transport of immune complexes across epithelial cells: a local defense function for IgA.
The polymeric immunoglobulin receptor (pIgR) on mucosal epithelial cells binds dimeric IgA (dIgA) on the basolateral surface and mediates transport of dIgA to the apical surface. Using Madin-Darby canine kidney epithelial cells stably transfected with pIgR cDNA, we found that soluble immune complexes (ICs) of 125I-labeled rat monoclonal antidinitrophenyl (DNP) dIgA (125I-dIgA) and DNP/biotin-bovine serum albumin were transported from the basolateral to the apical surface and then released. Monomeric IgA ICs were not transported, consistent with the specificity of pIgR for polymeric immunoglobulins. Essentially all the 125I-dIgA in apical culture supernatants was streptavidin precipitable, indicating that dIgA remained bound to antigen during transcytosis. While both dIgA and dIgA ICs bound pIgR with equal affinity (Kd approximately 8 nM), the number of high-affinity binding sites per cell was 2- to 3-fold greater for dIgA than for dIgA ICs. The extent of endocytosis of dIgA and dIgA ICs was correlated with the number of high-affinity binding sites. SDS/PAGE analysis of intracellular dIgA and dIgA ICs demonstrated that in both cases IgA remained undegraded during transport. The results suggest that the pathways of epithelial transcytosis of free dIgA and dIgA ICs are the same. Given the high population density of mucosal IgA plasma cells and the enormous surface area of pIgR-expressing mucosal epithelium, it is likely that significant local transcytosis of IgA ICs occurs in vivo. Such a process would allow direct elimination of IgA ICs at the mucosal sites where they are likely to form, thus providing an important defense function for IgA.


Dermatologist-level classification of skin cancer with deep neural networks.
Skin cancer, the most common human malignancy, is primarily diagnosed visually, beginning with an initial clinical screening and followed potentially by dermoscopic analysis, a biopsy and histopathological examination. Automated classification of skin lesions using images is a challenging task owing to the fine-grained variability in the appearance of skin lesions. Deep convolutional neural networks (CNNs) show potential for general and highly variable tasks across many fine-grained object categories. Here we demonstrate classification of skin lesions using a single CNN, trained end-to-end from images directly, using only pixels and disease labels as inputs. We train a CNN using a dataset of 129,450 clinical images-two orders of magnitude larger than previous datasets-consisting of 2,032 different diseases. We test its performance against 21 board-certified dermatologists on biopsy-proven clinical images with two critical binary classification use cases: keratinocyte carcinomas versus benign seborrheic keratoses; and malignant melanomas versus benign nevi. The first case represents the identification of the most common cancers, the second represents the identification of the deadliest skin cancer. The CNN achieves performance on par with all tested experts across both tasks, demonstrating an artificial intelligence capable of classifying skin cancer with a level of competence comparable to dermatologists. Outfitted with deep neural networks, mobile devices can potentially extend the reach of dermatologists outside of the clinic. It is projected that 6.3 billion smartphone subscriptions will exist by the year 2021 (ref. 13) and can therefore potentially provide low-cost universal access to vital diagnostic care.


Enhanced fungal specificity and in vivo therapeutic efficacy of a C-22-modified FK520 analog against C. neoformans.
Fungal infections are of mounting global concern, and the current limited treatment arsenal poses challenges when treating such infections. In particular, infections by Cryptococcus neoformans are associated with high mortality, emphasizing the need for novel therapeutic options. Calcineurin is a protein phosphatase that mediates fungal stress responses, and calcineurin inhibition by the natural product FK506 blocks C. neoformans growth at 37°C. Calcineurin is also required for pathogenesis. However, because calcineurin is conserved in humans and inhibition with FK506 results in immunosuppression, the use of FK506 as an anti-infective agent is precluded. We previously elucidated the structures of multiple fungal calcineurin-FK506-FKBP12 complexes and implicated the C-22 position on FK506 as a key point for differential modification of ligand inhibition of mammalian versus fungal target proteins. Through in vitro antifungal and immunosuppressive testing of FK520 (a natural analog of FK506) derivatives, we identified JH-FK-08 as a lead candidate for further antifungal development. JH-FK-08 exhibited significantly reduced immunosuppressive activity and both reduced fungal burden and prolonged survival of infected animals. JH-FK-08 exhibited additive activity in combination with fluconazole in vivo. These findings further advance calcineurin inhibition as an antifungal therapeutic approach. IMPORTANCE Fungal infections cause significant morbidity and mortality globally. The therapeutic armamentarium against these infections is limited, and the development of antifungal drugs has been hindered by the evolutionary conservation between fungi and the human host. With rising resistance to the current antifungal arsenal and an increasing at-risk population, there is an urgent need for the development of new antifungal compounds. The FK520 analogs described in this study display potent antifungal activity as a novel class of antifungals centered on modifying an existing orally active FDA-approved therapy. This research advances the development of much-needed newer antifungal treatment options with novel mechanisms of action.


EEG correlates of postural audio-biofeedback.
The control of postural sway depends on the dynamic integration of multi-sensory information in the central nervous system. Augmentation of sensory information, such as during auditory biofeedback (ABF) of the trunk acceleration, has been shown to improve postural control. By means of quantitative electroencephalography (EEG), we examined the basic processes in the brain that are involved in the perception and cognition of auditory signals used for ABF. ABF and Fake ABF (FAKE) auditory stimulations were delivered to 10 healthy naive participants during quiet standing postural tasks, with eyes-open and closed. Trunk acceleration and 19-channels EEG were recorded at the same time. Advanced, state-of-the-art EEG analysis and modeling methods were employed to assess the possibly differential, functional activation, and localization of EEG spectral features (power in α, β, and γ bands) between the FAKE and the ABF conditions, for both the eyes-open and the eyes-closed tasks. Participants gained advantage by ABF in reducing their postural sway, as measured by a reduction of the root mean square of trunk acceleration during the ABF compared to the FAKE condition. Population-wise localization analysis performed on the comparison FAKE - ABF revealed: (i) a significant decrease of α power in the right inferior parietal cortex for the eyes-open task; (ii) a significant increase of γ power in left temporo-parietal areas for the eyes-closed task; (iii) a significant increase of γ power in the left temporo-occipital areas in the eyes-open task. EEG outcomes supported the idea that ABF for postural control heavily modulates (increases) the cortical activation in healthy participants. The sites showing the higher ABF-related modulation are among the known cortical areas associated with multi-sensory, perceptual integration, and sensorimotor integration, showing a differential activation between the eyes-open and eyes-closed conditions.


Improved functional outcomes following exercise rehabilitation in patients with intermittent claudication.
The purposes of this study were to identify predictors of increased claudication distances following exercise rehabilitation in peripheral arterial occlusive disease (PAOD) patients with intermittent claudication and determine whether improved claudication distances translated into increased free-living daily physical activity in the community setting. Sixty-three patients were recruited (age, 68+/-1 years, mean +/- standard error). Patients were characterized on treadmill claudication distances, walking economy, peripheral circulation, cardiopulmonary function, self-perceived ambulatory function, body composition, baseline comorbidities, and free-living daily physical activity before and after a 6-month treadmill exercise program. Exercise rehabilitation increased distance to onset of claudication pain by 115% (178+/-22 m to 383+/-34 m; p < .001) and distance to maximal claudication pain by 65% (389+/-29 m to 641+/-34 m; p < .001). The increased distance to onset of pain was independently related to a 27% increase in calf blood flow (r = .42, p < .001) and to baseline age (r = -.26, p < .05), and the increased distance to maximal pain was predicted by a 10% increase in peak oxygen uptake (r = .41, p < .001) and by a 10% improvement in walking economy (r = -.34, p < .05). Free-living daily physical activity increased 31% (337+/-29 kcal/day to 443+/-37 kcal/day; p < .001) and was related to the increases in treadmill distances to onset (r = .24, p < .05) and to maximal pain (r = .45, p < .001). Increased claudication distances following exercise rehabilitation are mediated through improvements in peripheral circulation, walking economy, and cardiopulmonary function, with younger patients having the greatest absolute ambulatory gains. Furthermore, improved symptomatology translated into enhanced community-based ambulation.


Recognition Tunneling of Canonical and Modified RNA Nucleotides for Their Identification with the Aid of Machine Learning.
In the present study, we demonstrate a tunneling nanogap technique to identify individual RNA nucleotides, which can be used as a mechanism to read the nucleobases for direct sequencing of RNA in a solid-state nanopore. The tunneling nanogap is composed of two electrodes separated by a distance of <3 nm and functionalized with a recognition molecule. When a chemical entity is captured in the gap, it generates electron tunneling currents, a process we call recognition tunneling (RT). Using RT nanogaps created in a scanning tunneling microscope (STM), we acquired the electron tunneling signals for the canonical and two modified RNA nucleotides. To call the individual RNA nucleotides from the RT data, we adopted a machine learning algorithm, support vector machine (SVM), for the data analysis. Through the SVM, we were able to identify the individual RNA nucleotides and distinguish them from their DNA counterparts with reasonably high accuracy. Since each RNA nucleoside contains a hydroxyl group at the 2'-position of its sugar ring in an RNA strand, it allows for the formation of a tunneling junction at a larger nanogap compared to the DNA nucleoside in a DNA strand, which lacks the 2' hydroxyl group. It also proves advantageous for the manufacture of RT devices. This study is a proof-of-principle demonstration for the development of an RT nanopore device for directly sequencing single RNA molecules, including those bearing modifications.


Humic Substances Contribute to Plant Iron Nutrition Acting as Chelators and Biostimulants.
Improvement of plant iron nutrition as a consequence of metal complexation by humic substances (HS) extracted from different sources has been widely reported. The presence of humified fractions of the organic matter in soil sediments and solutions would contribute, depending on the solubility and the molecular size of HS, to build up a reservoir of Fe available for plants which exude metal ligands and to provide Fe-HS complexes directly usable by plant Fe uptake mechanisms. It has also been shown that HS can promote the physiological mechanisms involved in Fe acquisition acting at the transcriptional and post-transcriptional level. Furthermore, the distribution and allocation of Fe within the plant could be modified when plants were supplied with water soluble Fe-HS complexes as compared with other natural or synthetic chelates. These effects are in line with previous observations showing that treatments with HS were able to induce changes in root morphology and modulate plant membrane activities related to nutrient acquisition, pathways of primary and secondary metabolism, hormonal and reactive oxygen balance. The multifaceted action of HS indicates that soluble Fe-HS complexes, either naturally present in the soil or exogenously supplied to the plants, can promote Fe acquisition in a complex way by providing a readily available iron form in the rhizosphere and by directly affecting plant physiology. Furthermore, the possibility to use Fe-HS of different sources, size and solubility may be considered as an environmental-friendly tool for Fe fertilization of crops.


Increased fetal iron load in rhesus hemolytic disease.
There is little information about the iron overload caused by hemolysis in fetuses affected with rhesus hemolytic disease (RHD). The authors therefore studied the iron load in RHD by measuring cord blood ferritin levels in babies affected with RHD and gestational age- and weight-matched controls. Cord blood ferritin levels were higher in babies with RHD. Intrauterine transfusions did not affect the ferritin status of the babies with RHD and there was no correlation between hemoglobin and ferritin levels. The results indicate that there is an increased intrauterine iron load in babies with RHD, independent from intrauterine transfusions and rate of hemolysis.


Variations in the application of exception from informed consent in a multicenter clinical trial.
Exception from infor med consent (EFIC) is allowed using federal regulations 21 CFR 50.24 and facilitates research on patients with critical conditions such as cardiac arrest. Little is known regarding the differences in the application of EFIC requirements such as community consultation (CC), public disclosure (PD) and patient notification. We sought to characterize variations in the fulfillment of EFIC requirements in a national multicenter clinical trial in the United States. We determined the strategies for fulfillment of EFIC requirements at five regional coordinating centers of the Pragmatic Airway Resuscitation Trial (PART), a cluster-crossover randomized trial comparing airway devices in out-of-hospital cardiac arrest. We collected information from the including site demographics, how CC and PD were implemented, methods undertaken by the site investigative team to meet the local IRB's interpretation, and patient notification timing (post-enrollment). We analyzed the data using descriptive statistics. Sites had multiple approaches to CC, including social media advertising, random digit dialing surveys, working with city officials, and websites with embedded surveys. All sites used more than one approach for conducting CC. Public Disclosure activities included press releases through various means, website documentation, and letters to community members and local officials. Time from CC to study approval ranged from 42 days to 253 days. EFIC implementation varies across sites and highlight community and regional variation. Different EFIC approaches may be needed to effectively accomplish the goals of community consultation, public disclosure, and patient notification.


Understanding acceptance of contactless monitoring technology in home-based dementia care: a cross-sectional survey study among informal caregivers.
There is a growing interest to support home-based dementia care via contactless monitoring (CM) technologies which do not require any body contact, and allow informal caregivers to remotely monitor the health and safety of people with dementia (PwD). However, sustainable implementation of CM technologies requires a better understanding of informal caregivers' acceptance. This study aimed to examine the (1) general acceptance of CM technology for home-based dementia care, (2) acceptance of different sensor types and use scenarios, and (3) differences between accepters and refusers of CM technology. A cross-sectional online survey was conducted among n = 304 informal caregivers of community-dwelling PwD [Mean(SD) age = 58.5 (10.7)] in the Netherlands and Germany. The survey contained a textual and graphical introduction to CM technologies, as well as questions targeting (1) general acceptance of CM technology, (2) acceptance of seven different contactless sensor types, (3) acceptance of five different use scenarios, and (4) caregivers' own and their care recipients' personal characteristics. Data were examined using descriptive and bivariate analyses. Participants' general acceptance of CM technology was slightly positive. We found significant differences in acceptability between contactless sensor types (p < .001). RF-based sensors (e.g., radar) and light sensors were considered most acceptable, whereas camera-based sensors and audio sensors (e.g., microphones, smart speakers) were seen as least acceptable for home-based dementia care. Furthermore, participants' acceptance of different use scenarios for CM technology varied significantly (p < .001). The intention to use CM technology was highest for detecting emergencies (e.g., falls, wandering), and lowest for predicting acute situations (e.g., fall prediction). Lastly, accepters and refusers of CM technology significantly differed regarding gender (p = .010), their relation with the PwD (p = .003), eHealth literacy (p = .025), personal innovativeness (p < .001), usage of safety technology (p = .002), and the PwD's type of cognitive impairment (p = .035) and housing situation (p = .023). Our findings can inform the development and implementation of acceptable CM technology to support home-based dementia care. Specifically, we show which sensor types and use scenarios should be prioritized from the informal caregiver's view. Additionally, our study highlights several personal characteristics associated with informal caregivers' acceptance of CM technology that should be taken into account during implementation.


Evaluation of skin cancer screening in Germany - participation, tumor detection and interval tumors based on SHI data.
The rising incidence of skin cancer in Germany has increased the need for secondary prevention measures. For this purpose, a statutory skin cancer screening for insured persons aged 35 and older was introduced on 1 June 2008. The aim of this work package in the Innovation Fund project "Perspectives of a multimodal evaluation of early skin cancer detection" (Pertimo) was to test an evaluation of skin cancer screening using secondary data. The data basis was statutory insured persons of the DAK Health from the age of 35 who were insured as of 31 December 2010 and were followed up until the end of 2015. The rates of participation, skin tumors detected in skin cancer screening (tumor detections), and interval tumors that occurred within two years after a finding-free skin cancer screening were calculated. The biennial skin cancer screening take-up rate in 2014 and 2015 was 33.6% for women and 32.6% for men. Of those screened, 4.2% had a skin cancer finding (tumor detection) in the course of skin cancer screening. Of all incident skin cancer diagnoses (2012-2015), 50.1% were detected in skin cancer screening. In 1.5% of the insured persons with skin cancer screening without findings, an incidental skin tumor was diagnosed in the following two years (interval tumor). The data from the statutory health insurance mapped the skin cancer screening occurrence in Germany and highlighted the importance of dermatologists in the screening process. The analysis provided important new insights.


Short communication. Characterization of chloroplast region rrn16-rrn23S from the tropical timber tree Cedrela odorata L. and de novo construction of a transplastomic expression vector suitable for Meliaceae trees and other economically important crops.
The forest tree Spanish cedar (Cedrela odorata L.) is well-known for its high-value timber; however, this species is attacked by the shoot borer (Hypsipyla grandella) during its early years of development, resulting in branched stems and making the plants useless for high-quality wood production. The generation of resistant varieties expressing entomotoxic proteins may be an alternative to pesticide treatments. The use of plastid transformation rather than nuclear transformation should be used because it reduces the risk of transgene dissemination by pollen. Chloroplast transformation vectors require an expression cassette flanked by homologous plastid sequences to drive plastome recombination. Thus, C. odorata plastome sequences are a prerequisite. The rrn16-rrn23 plastome region was selected, cloned, and characterized. When the sequence identity among the rrn16-rrn23 regions from C. odorata and Nicotiana tabacum was compared, 3 inDels of 240, 104, and 39 bp were found that might severely affect transformation efficiency. Using this region, a new transformation vector was developed using pUC19 as a backbone by inserting the rrn16-trnI and trnA-rrn23 sequences from C. odorata and adding 2 independent expression cassettes into the trnI-trnA intergenic region, conferring spectinomycin resistance, the ability to express the gfp reporter gene, and a site that can be used to express any other gene of interest.


Lipomatous lesions of the parotid gland. Potential pitfalls in fine needle aspiration biopsy diagnosis.
Nine cases of benign fatty tumors of the parotid studied initially by fine needle aspiration (FNA) biopsy and subsequently diagnosed by open biopsy were reviewed. All lesions presented as enlargements of the parotid gland. The cytologic diagnoses were correct in only five of the nine cases. While four of six FNA samples from lipomas were correctly diagnosed by cytology, the other two samples were classified as inadequate. Three cases of diffuse infiltration of the parotid by mature-appearing adipose tissue were cytologically characterized as "no diagnostic change" in two cases and sialosis in one case. The clinical, radiologic, cytologic and histologic features of these tumors are described to increase the familiarity of cytologists with these lesions, which may prevent unnecessary surgical intervention in some patients.


Steroidogenic factor 1 messenger ribonucleic acid expression in steroidogenic and nonsteroidogenic human tissues: Northern blot and in situ hybridization studies.
Steroidogenic factor-1 (SF-1), a tissue-specific orphan nuclear receptor, regulates the genes of several steroidogenic enzymes, Mullerian inhibiting substance, and the gonadotrophins. Also, this transcription factor is crucial for hypothalamic, adrenal, and gonadal organogenesis in the mouse. We recently cloned the human SF-1 (hSF-1) complementary DNA (cDNA) and now report the distribution of this factor's messenger RNA (mRNA) in human tissues. Northern blot analyses of peripheral tissues revealed high hSF-1 mRNA expression in the adrenal cortex and the gonads, but no hSF-1 mRNA was detected in the placenta. High hSF-1 mRNA expression also was seen in the spleen. In this tissue, in addition to the main transcript of 3.5-4 kb seen in the adrenal and gonads, two additional transcripts of 4.4 kb and 8 kb were noted. The additional 4.4-kb transcript also was seen in several peripheral tissues and various components of the brain. However, adult liver and heart showed only the 4.4-kb transcript. In the human brain, hSF-1 mRNA expression was widespread, including several components of the limbic system. In situ hybridization studies confirmed the strong expression of hSF-1 mRNA in adrenal cortex, ovary, testis, and the spleen, primarily within reticuloendothelial cells. Thus, in the human, the hSF1 mRNA is present in both steroidogenic and nonsteroidogenic tissues, albeit not in the placenta. In the central nervous system, the expression of hSF-1 mRNA is widespread. It is composed of several different mRNA species distributed in a tissue-specific fashion. These findings suggest that hSF-1 may play a role in reticuloendothelial/immune cell maturation and/or function, as well as nervous system development and/or neurosteroid biosynthesis.


The Prognostic and Functional Impact of Sprouty 2 Expression in Non-small Cell Lung Cancer.
We represent Sprouty 2 (Spry2) expression analysis and its association with key driver mutations and clinical features of patients with non-small cell lung cancer as the largest ex vivo data. The strength of Spry2 expression was evaluated using the immunoreactivity score (IRS), which was calculated using the following formula: IRS=(staining intensity score) SI×(percentage of positively stained cells) PP. The median IRS score was defined as the cutoff value. Patients were grouped as "weak immunoreactivity score" (IRS: 0 to 4) or "strong immunoreactivity score" (IRS: ≥4) with respect to the IRS score. The intensity and percentage of Spry2 staining were significantly lower in tumor tissues than in normal lung tissues ( P <0.0001). Patients' characteristics were similar for both groups, except for smoking status and, brain and lymph node metastasis. Overall survival of patients with a strong immunoreactivity score was significantly lower than those with a weak immunoreactivity score among metastatic patients (6.9 mo vs. 13.6, P =0.023) and adenocarcinoma histology (7.0 mo vs. not reached, P =0.003). Spry2 expression was lower in tumor tissues than in normal lung parenchyma. Increased expression of Spry2 is associated with poor prognosis. There were no significant associations between epidermal growth factor receptor, anaplastic lymphoma kinase, or c-ros oncogene 1 rearrangement and Spry2 expression. Despite the absence of KRAS mutational analysis, the clinical and epidemiological features of patients suggested that KRAS mutation might be an underlying determinant factor of the functional role of Spry2 in non-small cell lung cancer.


In situ Ca2+ imaging of the enteric nervous system.
Reflex behaviors of the intestine are controlled by the enteric nervous system (ENS). The ENS is an integrative network of neurons and glia in two ganglionated plexuses housed in the gut wall. Enteric neurons and enteric glia are the only cell types within the enteric ganglia. The activity of enteric neurons and glia is responsible for coordinating intestinal functions. This protocol describes methods for observing the activity of neurons and glia within the intact ENS by imaging intracellular calcium (Ca(2+)) transients with fluorescent indicator dyes. Our technical discussion focuses on methods for Ca(2+) imaging in whole-mount preparations of the myenteric plexus from the rodent bowel. Bulk loading of ENS whole-mounts with a high-affinity Ca(2+) indicator such as Fluo-4 permits measurements of Ca(2+) responses in individual neurons or glial cells. These responses can be evoked repeatedly and reliably, which permits quantitative studies using pharmacological tools. Ca(2+) responses in cells of the ENS are recorded using a fluorescence microscope equipped with a cooled charge-coupled device (CCD) camera. Fluorescence measurements obtained using Ca(2+) imaging in whole-mount preparations offer a straightforward means of characterizing the mechanisms and potential functional consequences of Ca(2+) responses in enteric neurons and glial cells.


Cytochrome P-450-mediated metabolism of the individual enantiomers of the antidepressant agent reboxetine in human liver microsomes.
In vitro studies were conducted to identify the hepatic cytochrome P-450 (CYP) enzymes responsible for the oxidative metabolism of the individual enantiomers of reboxetine. In human liver microsomes, each reboxetine enantiomer was metabolized to one primary metabolite, O-desethylreboxetine, and three minor metabolites, two arising via oxidation of the ethoxy aromatic ring and a third yet unidentified metabolite. Over a concentration range of 2 to 200 microM, the rate O-desethylreboxetine formation for either enantiomer conformed to monophasic Michaelis-Menten kinetics. Evidence for a principal role of CYP3A in the formation of O-desethylreboxetine for (S, S)-reboxetine and (R,R)-reboxetine was based on the results from the following studies: 1) inhibition of CYP3A activity by ketoconazole markedly decreased the formation of O-desethylreboxetine, whereas inhibitors selective for other CYP enzymes did not inhibit reboxetine metabolism, 2) formation of O-desethylreboxetine correlated (r(2) = 0.99; p <.001) with CYP3A-selective testosterone 6-beta-hydroxylase activity across a population of human livers (n = 14). Consistent with inhibition and correlation data, O-desethylreboxetine formation was only detectable in incubations using microsomes prepared from a Baculovirus-insect cell line expressing CYP3A4. Furthermore, the apparent K(M) for the O-desethylation of reboxetine in cDNA CYP3A4 microsomes was similar to the affinity constants determined in human liver microsomes. In addition, (S,S)-reboxetine and (R,R)-reboxetine were found to be competitive inhibitors of CYP2D6 and CYP3A4 (K(i) = 2.5 and 11 microM, respectively). Based on the results of the study, it is concluded that the metabolism of both reboxetine enantiomers in humans is principally mediated via CYP3A.


In utero exposure to environmentally relevant concentrations of PCB 153 and PCB 118 disrupts fetal testis development in sheep.
Polychlorinated biphenyls (PCB) are environmental pollutants linked to adverse health effects including endocrine disruption and disturbance of reproductive development. This study aimed to determine whether exposure of pregnant sheep to three different mixtures of PCB 153 and PCB 118 affected fetal testis development. Ewes were treated by oral gavage from mating until euthanasia (d 134), producing three groups of fetuses with distinct adipose tissue PCB levels: high PCB 153/low PCB 118 (n = 13), high PCB 118/low PCB 153 (n = 14), and low PCB 153/low PCB 118 (n = 14). Fetal testes and blood samples were collected for investigation of testosterone, testis morphology, and testis proteome. The body weight of the offspring was lower in the high PCB compared to the low PCB group, but there were no significant differences in testis weight between groups when corrected for body weight. PCB exposure did not markedly affect circulating testosterone. There were no significant differences between groups in number of seminiferous tubules, Sertoli cell only tubules, and ratio between relative areas of seminiferous tubules and interstitium. Two-dimensional (2D) gel-based proteomics was used to screen for proteomic alterations in the high exposed groups relative to low PCB 153/low PCB 118 group. Twenty-six significantly altered spots were identified by liquid chromatography (LC)-mass spectroscopy (MS)/MS. Changes in protein regulation affected cellular processes as stress response, protein synthesis, and cytoskeleton regulation. The study demonstrates that in utero exposure to different environmental relevant PCB mixtures exerted subtle effects on developing fetal testis proteome but did not significantly disturb testis morphology and testosterone production.


Power Doppler ultrasound mapping in nonobstructive azoospermic patients prior to testicular sperm extraction.
This study was conducted to localize the testicular regions, which have better blood circulation by power Doppler ultrasonography in patients with nonobstructive azoospermia before testicular sperm extraction (TESE), and to investigate whether these vascularized areas have a high sperm retrieval rate or not. We evaluated 110 testes of 55 cases that were diagnosed as nonobstructive azoospermia. The mean age of the study group was 33 years (range 26 to 42). Patients with Y chromosome microdeletions, karyotype and hormonal abnormalities (except elevated FSH levels) were excluded from the study. In all cases, testes were evaluated by power Doppler ultrasonography before testicular sperm extraction. Testis was divided vertically into five equal parts and the area with maximum vascularity was determined subjectively. During testicular sperm extraction, starting from best-perfused areas, biopsies were done. If no motile or sufficient amount of sperm was found, TESE procedure was tried on the contralateral testis. TESE were performed from 82 testes and for the regions that show good and poor vascularity. The sperm finding rate was 38% and 14%, respectively (OR = 3.55)(p = 0.001). Power Doppler ultrasound mapping of the testis in nonobstructive azoospermic cases is a reliable and informative method to assess spermatogenic foci. It is a noninvasive technique that minimizes the unnecessary removal of hormone producing tissue and gives chance to end the TESE earlier than currently practiced procedures.


[Estrogen treatment for prevention and therapy of osteoporosis].
Postmenopausal osteoporosis affects 30% of all women. Major risk factors include hereditary factors, deficiency of calcium and vitamin D in the diet, too little exercise, excessive alcohol consumption and a reduction in the amount or duration of sex hormone secretion (late menarche, early climacterium, etc.). Osteoporosis prophylaxis is of great importance. Since in the early stages, osteoporosis does not produce symptoms, an early diagnostic work-up involving osteodensitometry makes good sense in patients with individual risk factors. In addition to physical activity, basic countermeasures always include adequate calcium and vitamin D supplementation. Replacement therapy with estrogens (usually in combination with gestagens) is an effective and causal treatment of postmenopausal osteoporosis. However, the indication must be established individually on the basis of a benefit-risk consideration.


Comparing whole-genome sequencing with Sanger sequencing for spa typing of methicillin-resistant Staphylococcus aureus.
spa typing of methicillin-resistant Staphylococcus aureus (MRSA) has traditionally been done by PCR amplification and Sanger sequencing of the spa repeat region. At Hvidovre Hospital, Denmark, whole-genome sequencing (WGS) of all MRSA isolates has been performed routinely since January 2013, and an in-house analysis pipeline determines the spa types. Due to national surveillance, all MRSA isolates are sent to Statens Serum Institut, where the spa type is determined by PCR and Sanger sequencing. The purpose of this study was to evaluate the reliability of the spa types obtained by 150-bp paired-end Illumina WGS. MRSA isolates from new MRSA patients in 2013 (n = 699) in the capital region of Denmark were included. We found a 97% agreement between spa types obtained by the two methods. All isolates achieved a spa type by both methods. Nineteen isolates differed in spa types by the two methods, in most cases due to the lack of 24-bp repeats in the whole-genome-sequenced isolates. These related but incorrect spa types should have no consequence in outbreak investigations, since all epidemiologically linked isolates, regardless of spa type, will be included in the single nucleotide polymorphism (SNP) analysis. This will reveal the close relatedness of the spa types. In conclusion, our data show that WGS is a reliable method to determine the spa type of MRSA.


Strain-induced switching of heat current direction generated by magneto-thermoelectric effects.
Since the charge current plays a major role in information processing and Joule heating is inevitable in electronic devices, thermal management, i.e., designing heat flows, is required. Here, we report that strain application can change a direction of a heat current generated by magneto-thermoelectric effects. For demonstration, we used metallic magnets in a thin-film form, wherein the anomalous Ettingshausen effect mainly determines the direction of the heat flow. Strain application can alter the magnetization direction owing to the magnetoelastic effect. As a result, the heat current, which is in the direction of the cross product of the charge current and the magnetization vector, can be switched or rotated simply by applying a tensile strain to the metallic magnets. We demonstrate 180° switching and 90° rotation of the heat currents in an in-plane magnetized Ni sample on a rigid sapphire substrate and a perpendicularly magnetized TbFeCo film on a flexible substrate, respectively. An active thermography technique was used to capture the strain-induced change in the heat current direction. The method presented here provides a novel method for controlling thermal energy in electronic devices.


2-O-methyl-D-glucuronic acid, a new hexuronic acid of biological origin.
Acid hydrolysis of the extracellular polysaccharide of Porphyridium cruentum (a unicellular red alga) produced a mixture of aldobiuronic acids and free hexuronic acids. Fractionation of this mixture on an ion-exchange column yielded a hexuronic acid characterized as the title compound. Its identity was confirmed by chromatographic comparisons with the authentic compound, by reduction to the corresponding methylated aldose, by resistance to controlled lead tetra-acetate oxidation and by chemical-ionization mass spectrometry. Complete spectra have been deposited as Supplementary Publication SUP50062 (7 pages) with the British Library (Lending Division), Boston Spa, Wetherby, W. Yorkshire LS23 7BQ, U.K., from whom copies may be obtained under the terms given in Biochem. J. (1976) 153, 5.


['Smart drugs' enticements on the Internet].
'Smart drugs' are drugs supposed to enhance certain physical or mental functions, without harmful side effects. The target group for the misleading advertisement campaigns on the Net is mostly the younger generation. Smart drugs clearly are pharmacologically active: phenytoin, selegilin and growth hormone are examples of compounds that can be ordered from mail order pharmacies without prescription. Misleading medical information on the Internet is undesirable and potentially harmful and the widespread availability of smart drugs should be realised by pharmacists and physicians. Screening the Net for this kind of information should be implemented as a standard procedure and specific, targeted information to put these drugs in a realistic perspective could be the first step leading to 'debunking' the Internet.


Restricting conversations about voluntary assisted dying: implications for clinical practice.
On 19 June 2019, assisted dying became lawful in Victoria, the second most populous state in Australia. Section 8 of the Voluntary Assisted Dying Act is a legislative safeguard that is designed to ensure a patient's request for assistance to die is voluntary. This section prohibits health practitioners from initiating a conversation about assisted dying with the patient. This article explores the potential implications of this prohibition for effective communication between doctors and their patients, and the ability of doctors to provide high quality end-of-life (EOL) care in some cases. The authors reviewed and analysed literature on the importance of communication at the EOL including the need to understand and appropriately respond to Desire to Die or Desire to Hasten Death statements. A legal critique of section 8 of the Victorian Voluntary Assisted Dying Act was also undertaken to determine the scope of this new duty and how it aligns with existing legal obligations that would otherwise require doctors to provide information about EOL options requested by a patient. Contemporary literature suggests that open and honest communication between doctor and patient including the provision of information about all EOL options when sought by the patient represents good clinical practice and will lead to optimal EOL care. The provision of such information also reflects professional, ethical and legal norms. Despite (arguably) promoting an appropriate policy objective, the legislative prohibition on health professionals initiating conversations about voluntary assisted dying may, in cases where patients seek information about all EOL options, lead to less optimal patient outcomes.


Detection of t(14;18) in turkish follicular lymphomas using the polymerase chain reaction.
A t(14;18) translocation is closely associated with the follicular lymphoma but is also seen in diffuse B cell lymphomas with a previous history of a follicular lymphoma as well as de novo diffuse lymphomas. Estimation of the frequency of t(14;18) in follicular lymphoma vary widely from 33 to 89%. Furthermore, no extensive data have been published on the frequency of t(14;18) in Turkish cases of follicular lymphoma. Representative tissue blocks from 67 patients with follicular lymphoma, 12 cases of diffuse large B cell lymphomas and 11 cases of reactive hyperplasias were examined for the presence of this translocation using PCR. DNA probes capable of detecting rearrangement at both the major and minor break point regions were employed. We could detect t(14;18) in 46 out of 67 cases (68.7%) of follicular and 25% of diffuse large B cell lymphomas. In follicular lymphomas 64.2% of these break points were at mbr and 4.5% were at the mcr region. Review of the literature showed that comparable results have been obtained previously using molecular techniques. Our data showed that despite the relative infrequency of follicular lymphomas in the Turkish population these lymphomas share a common molecular pathogenesis with involvement of bcl-2 gene and background incidence of such rearrangement is similar in all populations, regardless the incidence of folicular lymphoma.


Female genital mutilation/cutting: changes and trends in knowledge, attitudes, and practices among health care professionals in The Gambia.
Female genital mutilation/cutting (FGM/C) is a harmful traditional practice that affects two out of three girls in The Gambia, seriously threatening their life and well-being with severe health consequences. By tracking the reference values established in former research conducted between 2009 and 2011, the objectives of this study are to explore trends and to measure and assess changes in knowledge, attitudes, and practices regarding FGM/C among health care professionals (HCPs) in The Gambia. A cross-sectional descriptive study was designed to collect and analyze data from an overall stratified sample consisting of 1,288 HCPs including health professionals and students throughout the six regions of The Gambia. Data were collected by the implementation of a self-administered written knowledge, attitudes, and practices questionnaire between 2012 and 2014. The results of this study showed that 76.4% of HCPs are eager to abandon FGM/C, and 71.6% of them regard it as a harmful practice with negative consequences on life and health. HCPs reported more knowledge and favorable attitudes towards FGM/C abandonment, being better able to identify the practice, more aware of its health complications, and more concerned in their essential role as social agents of change. However, 25.4% of HCPs still embraced the continuation of the practice, 24.4% expressed intention of subjecting their own daughters to it, and 10.5% declared to have performed it within their professional praxis. Findings confirm progress in knowledge and attitudes regarding FGM/C among HCPs, who are better skilled to understand and manage the consequences. Nevertheless, discrepancies between information, intention, and behavior unveil resistance in practice and proves that FGM/C medicalization is increasing. Thus, there is an urgent need to support HCPs in the integration of FGM/C preventive interventions within the public health system, to address arguments favoring medicalization, and to use data to design appropriate strategies.


BLV proviral DNA in the genome of the target lymphocyte.
Integration of bovine leukemia proviral DNA in the genome of infected cells was investigated in cattle affected by either the persistent lymphocytosis or the lymph node tumor form of enzootic bovine leukosis. In persistent lymphocytosis, proviral DNA was found to be integrated at a large number of genomic sites in one-fourth to one-third of circulating leucocytes. In the lymph node tumor form, in contrast, proviral DNA was found to be integrated at one or very few sites in the genomes of a larger fraction of both circulating leucocytes and lymph node tumor cells.


The cognitive and behavioral effects of D-amphetamine and nicotine sensitization in adult zebrafish.
Zebrafish are growing in use as a model for understanding drug dependence and addiction. Sensitization paradigms have been a useful tool in identifying mechanisms involved in drug-induced behavioral and neurological changes, but in zebrafish have tended to focus on locomotor, rather than cognitive, endpoints. Here, we used a novel method, the FMP Y-maze, which measures continuous performance through a series of repeated binary choices (L vs R), to establish a model for assessing parameters associated with psychostimulant-induced behavioral and cognitive sensitization in adult zebrafish. Repeat, intermittent exposure to d-amphetamine (AMPH) for 14 days increased alternations (LRLR) in the maze, suggesting improved working memory, which was enhanced further following drug challenge after a short withdrawal period, suggesting behavioral sensitization. However, this cognitive enhancement coincided with a reduction in the use of other exploration strategies, hypolocomotion, and inhibition of cognitive flexibility. Like AMPH, exposure to nicotine (NIC) increased alternations following drug challenge after chronic treatment. Repeat NIC exposure appeared to induce both cognitive and psychomotor sensitization, as evidenced by increased working memory performance (alternations) and locomotor activity, without negatively impacting other search strategies or cognitive flexibility. Chronic treatment with AMPH or NIC boosts cognitive performance in adult zebrafish. Cognitive sensitization occurred with both drugs, resulting in enhanced working memory; however, repeat AMPH exposure, following a withdrawal period, resulted in inhibited cognitive flexibility, an effect not evident with repeat NIC exposure. Cognitive and behavioral sensitization paradigms in zebrafish could serve as a useful tool for assessing cognitive states which result in cognitive enhancing or impairing effects of drugs.


Recurrent massive ascites due to mossy endometriosis.
To report the medical and surgical management of a rare case of recurrent moss-like endometriosis and associated hemorrhagic ascites. Video description of the case, demonstration of the surgical technique, discussion of the histology, and review of endometriosis-associated ascites. Tertiary referral center. A 26-year-old nulliparous woman of Nigerian heritage with recurrent hemorrhagic ascites due to endometriosis. Three years previously she underwent an exploratory laparotomy for similar symptoms, and 7 L of hemorrhagic ascites were evacuated from her abdomen. Friable lesions covering the peritoneum of the uterus, bladder, and pouch of Douglas were biopsied and consistent with endometriosis. After her initial surgery, the patient was hormonally suppressed with goserelin for 3 months and oral medroxyprogesterone for 1 year. She then stopped the medications to attempt pregnancy but was unsuccessful. She used clomiphene for 3 months, and the ascites reaccumulated. The patient was started on depot leuprolide and oral norethindrone, but the ascites persisted. The patient underwent small-diameter laparoscopy using a multipuncture technique, evacuation of 7.8 L of hemorrhagic ascites, enterolysis, appendectomy, chromopertubation, and treatment of the endometriosis. Diffuse olive-green "mossy" endometriosis lesions blanketed the pelvic and abdominal peritoneum. The endometriosis was surgically resected with a combination of peritoneal stripping, excision with carbon dioxide laser, and ablation with neutral argon plasma. Examination of the ascites showed scattered hemosiderin-laden macrophages in a background of red blood cells. Histology of the olive-green mossy lesions revealed dense sheets of hemosiderin-laden macrophages and rare foci of endometriosis. Surgical reports in deidentified patients are exempted from Institutional Review Board approval. The patient gave consent to use photography and images for the video article. No postoperative hormone suppression was given to the patient because she desired pregnancy. At 6 months after her second surgery, the patient had not achieved pregnancy, but the ascites had not reaccumulated. She was referred for further infertility care. This rare form of mossy endometriosis often mimics ovarian cancer, pelvic tuberculosis, and other gynecologic conditions, but when identified, the endometriosis can be treated and symptoms can subside with drainage of the ascites, thorough ablation of the diffuse, superficial lesions, and restoration of anatomy.


Tissue inhibitor of metalloproteinase 1 (TIMP-1) may be an autocrine growth factor in scleroderma fibroblasts.
In scleroderma (systemic sclerosis, SSc), an autoimmune disorder in which excessive extracellular matrix is deposited in skin and internal organs, one of the suggested contributory factors to the development of fibrosis is a decrease in collagenase activity that may be related to levels of serum tissue inhibitors of metalloproteinases 1 (TIMP-1). We recently reported that the serum TIMP-1 levels in SSc patients were elevated compared with normal controls. To determine the biologic significance of TIMP-1 in SSc, we compared the proliferative effects of TIMP-1 between normal and SSc fibroblasts. TIMP-1 showed significant mitogenic activity for both normal and SSc fibroblasts. The mitogenic responses to TIMP-1 (33-100 ng/ml) in SSc fibroblasts, however, were significantly greater than those in normal controls and were completely neutralized in the presence of anti-TIMP-1 IgG. Moreover, anti-TIMP-1 IgG partially but significantly blocked the basal mitogenic activities of SSc fibroblasts. SSc fibroblasts produced increased amounts of TIMP-1 relative to normal fibroblasts, as confirmed by western blotting, ELISA, and RT-PCR techniques. In contrast, transforming growth factor beta1 (TGF-beta1) upregulated TIMP-1 production in normal fibroblasts but not in SSc fibroblasts with elevated spontaneous secretion of TIMP-1. These observations suggest that TIMP-1 may play an important role as an autocrine growth factor in the fibrotic process in SSc.


Surface Waters and Urban Brown Rats as Potential Sources of Human-Infective Cryptosporidium and Giardia in Vienna, Austria.
Cryptosporidium and Giardia are waterborne protozoa that cause intestinal infections in a wide range of warm-blooded animals. Human infections vary from asymptomatic to life-threatening in immunocompromised people, and can cause growth retardation in children. The aim of our study was to assess the prevalence and diversity of Cryptosporidium and Giardia in urban surface water and in brown rats trapped in the center of Vienna, Austria, using molecular methods, and to subsequently identify their source and potential transmission pathways. Out of 15 water samples taken from a side arm of the River Danube, Cryptosporidium and Giardia (oo)cysts were detected in 60% and 73% of them, with concentrations ranging between 0.3-4 oocysts/L and 0.6-96 cysts/L, respectively. Cryptosporidium and Giardia were identified in 13 and 16 out of 50 rats, respectively. Eimeria, a parasite of high veterinary importance, was also identified in seven rats. Parasite co-ocurrence was detected in nine rats. Rat-associated genotypes did not match those found in water, but matched Giardia previously isolated from patients with diarrhea in Austria, bringing up a potential role of rats as sources or reservoirs of zoonotic pathogenic Giardia. Following a One Health approach, molecular typing across potential animal and environmental reservoirs and human cases gives an insight into environmental transmission pathways and therefore helps design efficient surveillance strategies and relevant outbreak responses.


Using Demand Curves to Quantify the Reinforcing Value of Social and Solitary Drinking.
Young adults typically drink in social settings and report high levels of episodic heavy drinking despite a range of adverse consequences. Behavioral economics posits that this may reflect a reinforcer pathology in which alcohol is overvalued relative to other reinforcers. Theoretically, the value of alcohol is related to both the direct pharmacological effects of alcohol (euphoria, sedation) and the associated social reinforcement, but to date no studies have differentiated the value of social vs. solitary drinking. The current study examines two modified hypothetical alcohol purchase tasks (APTs), one explicitly social and one explicitly solitary, in order to quantify the reward value of social vs. solitary drinking and to determine whether there are unique clinical correlates of solitary alcohol demand. Participants were young adults (N = 274, Mage = 25.15, SD = 4.10) recruited from Mturk and from a university subject pool. Participants completed a solitary and social APT, in addition to measures of alcohol consumption and problems. Participants reported significantly greater demand in the social APT compared to the solitary APT across all demand indices. Elevated solitary and social demand were associated with elevated levels of alcohol use and problems. Using a residualized change approach, solitary demand amplitude (maximum consumption and expenditure) and persistence (price sensitivity) contributed additional variance above and beyond their social APT composite counterparts in predicting typical drinks per week and the self-care, academic/occupational, and physical dependence subscales of the YAACQ. The presence of peers increases alcohol demand compared to a solitary scenario, and greater relative solitary drinking demand may be a risk factor for greater alcohol consumption and problems.


Infection of cattle with Cryptosporidium parvum: mast cell accumulation in small intestine mucosa.
Mast cells might play an important role as the major effector cells in the immune response against Cryptosporidium parvum. C. parvum is a protozoan parasite that causes cryptosporidiosis in animals and humans worldwide. To investigate the interaction between C. parvum and mast cells during infection, nine 3-day-old male calves were orally challenged with 10(6) oocysts of C. parvum per calf. The distribution of mast cells in the mucosa of the small intestine was analyzed by toluidine blue staining. The concentrations of histamine and the cytokines interferon-γ, interleukin-4, interleukin-2, and interleukin-12 were measured in the serum, and the histamine levels were also determined from the intestinal contents. The following clinical signs were monitored: nausea, watery diarrhea, dehydration, and weight loss. Oocysts were shed in the feces during the infection period. C. parvum infection induced an increase in mast cell numbers in the mucosa of the small intestine in distinct temporal and spatial patterns. Infection with C. parvum can induce mastocytosis in the entire small intestinal mucosa in immune-competent calves, and the presence of the parasites influences the distribution profile of the mast cells.


The Effect of Dezocine on the Median Effective Dose of Sufentanil-Induced Respiratory Depression in Patients Undergoing Spinal Anesthesia Combined with Low-Dose Dexmedetomidine.
The application of sedation and analgesia in spinal anesthesia has many benefits, but the risk of respiratory depression (RD) caused by opioids cannot be ignored. We aimed to observe the effect of dezocine, a partial agonist of μ-receptor, on the median effective dose (ED50) of sufentanil-induced RD in patients undergoing spinal anesthesia combined with low-dose dexmedetomidine. Sixty-two patients were randomly assigned to dezocine group (DS) and control group (MS). After spinal anesthesia, mask oxygen (5 L/min) and dexmedetomidine (0.1 ug/kg) were given. Five minutes later, patients in the DS group received an Intravenous (IV) bolus of sufentanil and 0.05mg/kg dezocine, while patients in the MS group only received an IV bolus of sufentanil. ED50 of DS group was 0.342 ug/kg, 95% confidence interval (CI) was (0.269, 0.623) ug/kg, and the ED50 of MS group was 0.291 ug/kg, 95% CI was (0.257, 0.346) ug/kg. There was no difference in the type and treatment measures of RD and hemodynamic changes between the two groups, and no serious adverse reactions occurred in either group. Dezocine can improve RD induced by sufentanil in patients with spinal anesthesia combined with low-dose dexmedetomidine, and increase the safety window of sufentanil use.


[The solitary pulmonary nodule: the preliminary results in differential diagnosis by high-resolution computed tomography with a contrast medium].
High-resolution computed tomography (HRCT) with iodinated contrast material has been used by many authors to study solitary pulmonary nodules (SPNs). The degree of enhancement was correlated with the nodule malignancy. Forty adult patients were examined, before and after contrast agent administration, with incremental dynamic CT. We selected 22 patients with SPNs (3-30 mm phi, except one with 40 mm phi). The CT numbers of the inner nodule were calculated before and 1, 2 and 3 minutes after the i.v. administration of a weight-related dose (1.5 mL/kg/min) of nonionic iodinated contrast agent. A dose of 100 mL contrast agent was used in the first 6 patients. The difference in CT numbers between unenhanced images and the images with maximum enhancement (max. attenuation) was also calculated. Histologic diagnoses included 4 tuberculomas, 3 hamartomas and 15 malignant tumors (9 adenocarcinomas, 5 squamous cell carcinomas and 1 non-Hodgkin lymphoma). The CT numbers (in Hounsfield units, HU) of malignant nodules ranged 12-31 HU (mean: 21.5 HU) before contrast agent administration; the "long-standing" tuberculomas ranged 11-22 HU (mean: 16.5 HU) and the hamartomas had a mean density of 10.5 HU. We excluded for the study two "fresh" tuberculomas, one of which was surrounded by a low-attenuation infiltrate (the halo sign). We selected a threshold value of 20 HU on enhanced CT images to distinguish malignant (> or = 20 HU) from benign (< or = 20 HU) nodules. All lung cancers had complete enhancement (mean density: 35.5 HU). With 20 HU as the threshold value for a positive test, sensitivity was 100%, specificity 85.7%, positive predictive value 93.8% and negative predictive value 100%; test bias was 1.067. Positron emission tomography (PET) with 2-[fluorine-18] fluoro-2-deoxy-D glucose is reported to be as accurate as enhanced HRCT, but it does not provide accurate morphological information, is not widely available and it is quite expensive: therefore, in our opinion, CT should be preferred. After examining over 100 patients, we may use our results in the decision analysis comparing surgical risk with cancer risk.


Effect of a tailored leaflet to promote diabetic retinopathy screening among young adults with type 2 diabetes: a randomised controlled trial.
Young adults with type 2 diabetes (aged 18-39 years) are at risk of early onset and rapid progression of diabetic retinopathy, the leading cause of blindness and vision loss in working age adults. Early detection via retinal screening can prevent most vision loss, yet screening rates are consistently lower among this priority population than the general diabetes population. We aimed to test the effect of a tailored, evidence-based brief health behaviour change intervention (leaflet) on self-reported screening uptake, and previously identified social cognitive determinants of retinal screening. A pragmatic, two-arm randomised controlled trial was conducted from September 2014 to April 2015. Participants were stratified by prior screening uptake (Yes/No) and randomly allocated to intervention (leaflet) or 'usual care' control (no leaflet). Primary outcome was self-reported screening uptake four weeks post-intervention for 'No' participants who had not previously screened for diabetic retinopathy. Secondary outcome variables were changes in knowledge, attitudes, normative beliefs, intention and behavioural skills for all participants, irrespective of prior screening behaviour. To assess intervention effects on secondary outcome variables, we conducted independent samples t-tests (two-tailed) on pre-post change scores. 129 young adults (26% no prior retinal screen) completed baseline; 101 completed post-intervention. Power to determine effect on the primary outcome was curtailed by low recruitment of individuals with no prior retinal screen and loss to follow-up. Attrition was associated significantly with country of birth, language spoken at home, and marital status. Significant intervention effect was observed for one secondary outcome variable: knowledge of diabetic retinopathy (p = .03) with moderate effect (partial eta squared η2 = .05); no adverse effects were reported. Control group participants received the leaflet at study completion. This study confirms that a well-designed eye health and retinal screening promotion leaflet can increase knowledge of diabetic retinopathy, an important screening predictor. The study highlights the challenges of conducting 'real-world' health behaviour change research with this priority population, providing insights for clinicians and researchers. Strategies to recruit, engage and retain hard-to-reach populations are discussed including nonconventional alternatives to randomised controlled trial designs. ACTRN12614001110673, UTN No.: U1111-1161-9803. Registered 20 October 2014 - retrospectively registered https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=367127.


Issues of recruitment and maintaining high response rates in a longitudinal study of older hospital patients in England--pathways through care study.
To describe and discuss the methods used to recruit and maintain an unbiased sample of older discharged hospital patients in a study of the process and outcomes of hospital care. Prospective longitudinal interview study of consecutive patients admitted to hospital over a 12 month period and followed up for six months. Interviews took place in hospital five days after admission, at home 10 days after discharge, and six months after admission. Six hospital locations: three in the north of England and three in the south. People aged 65 and over admitted to hospital with a new stroke or fractured neck of femur, their significant other, and nursing staff caring for them. Of 3105 patients referred to the study, 2111 were eligible and 1671 (79%) were recruited. Recruited stroke patients were younger than those not recruited and rates differed between locations for both stroke and fractured neck of femur. By six months after admission 25% had died. Outcome data were obtained for 85% of the surviving patients. Patients who died were older and frailer before admission. Among survivors, outcome data for stroke patients were less likely to be obtained for men, those more able initially, and those who were married. Response rates to each interview differed according to respondent types. Interviews were more likely to be obtained with significant others than patients. Patients who were not able to be interviewed were older and frailer; significant others were less likely to be interviewed if the patients were younger and more able. High response rates can be achieved with very frail older people if strategies are adopted to maintain their interest and if self reported data are supplemented by interviewing significant others.


Effect of in ovo leptin administration on the development of Japanese quail.
Potential changes in the activity of endocrine axes related to growth as a result of leptin administration during embryonic development of birds were evaluated in the Japanese quail as a model bird with fast growth and development. On day 5 of incubation, 0.1 microg or 1 microg of recombinant mice leptin in 50 microl of phosphate buffered saline were injected into the albumen of eggs. Animals from each group were killed by decapitation on day 0, 2, 5, 7, 14, 21, 28, 35, 42, 49 and 56 of life. Plasma concentrations of triiodothyronine (T(3)), thyroxin (T(4)), corticosterone, testosterone, total lipids, triacylglycerols, cholesterol, glucose and alkaline phosphatase activity were measured. Quail treated by leptin hatched earlier (5-24 hours) and had a higher body weight than the control group (P<0.05-0.001). Mean body weight across the whole observed period was higher in both treated groups as compared to the control group (P<0.05). Leptin in ovo administration was accompanied by changes of endocrine and metabolic parameters during postembryonic development. The most prominent changes appeared immediately after hatching (T(3), T(4), total lipids, triacylglycerols) and before sexual maturity. It is suggested that leptin acts as a general signal of low energy status to neuroendocrine systems in birds which improves utilization of nutrients.


Cost-utility analysis of treatments for stage IB cervical cancer.
To analyze the cost-utility of two common clinical practices for stage IB cervical cancer patients from provider and societal viewpoints. A DECISION TREE MODEL WAS CONDUCTED TO EXAMINE VALUE FOR EXPENDITURE BETWEEN THE FOLLOWING: (1) radical hysterectomy with pelvic lymph node dissection (RHPLND) with or without postoperative adjuvant therapy according to the risk of recurrence and (2) concurrent chemoradiotherapy (CCRT). The relevant studies were identified to extract the probability data, and meta-analysis was performed. Direct medical costs were estimated from hospital database and medical records review. Direct non-medical costs and utility parameters were obtained through interviews with patients to estimate quality-adjusted life years (QALYs) outcome. The time horizon was according to the life expectancy of Thai women. From provider viewpoint, RHPLND and CCRT resulted in approximate costs of US $5,281 and US $5,218, respectively. The corresponding costs from societal viewpoint were US $6,533 and US $6,335, respectively. QALYs were 16.40 years for RHPLND and 15.94 years for CCRT. The estimated incremental cost effectiveness ratio of RHPLND in comparison to CCRT from provider and societal viewpoints were US $100/QALY and US $430/QALY, respectively. RHPLND had more cost-effectiveness than CCRT if patients did not need adjuvant therapy. The most effective parameter in model was a direct medical cost of CCRT. At the current ceiling ratio in Thailand, RHPLND provides better value for money than CCRT, with a probability of 75%. RHPLND is an efficient treatment for stage IB cervical cancer. This advantage is only for patients who require no adjuvant treatment.


Global Lysine Acetylome Analysis of LPS-Stimulated HepG2 Cells Identified Hyperacetylation of PKM2 as a Metabolic Regulator in Sepsis.
Sepsis-induced liver dysfunction (SILD) is a common event and is strongly associated with mortality. Establishing a causative link between protein post-translational modification and diseases is challenging. We studied the relationship among lysine acetylation (Kac), sirtuin (SIRTs), and the factors involved in SILD, which was induced in LPS-stimulated HepG2 cells. Protein hyperacetylation was observed according to SIRTs reduction after LPS treatment for 24 h. We identified 1449 Kac sites based on comparative acetylome analysis and quantified 1086 Kac sites on 410 proteins for acetylation. Interestingly, the upregulated Kac proteins are enriched in glycolysis/gluconeogenesis pathways in the Kyoto Encyclopedia of Genes and Genomes (KEGG) category. Among the proteins in the glycolysis pathway, hyperacetylation, a key regulator of lactate level in sepsis, was observed at three pyruvate kinase M2 (PKM2) sites. Hyperacetylation of PKM2 induced an increase in its activity, consequently increasing the lactate concentration. In conclusion, this study is the first to conduct global profiling of Kac, suggesting that the Kac mechanism of PKM2 in glycolysis is associated with sepsis. Moreover, it helps to further understand the systematic information regarding hyperacetylation during the sepsis process.


Proteomic Monitoring of B Cell Immunity.
Immune monitoring is critical in settings of infection, autoimmunity, and cancer, but our understanding of the diversity of the antibody immune repertoire has been limited to selected target antigens and epitopes. Development of new vaccines requires monitoring of B cell immunity and identification of candidate antigens. As vaccines become more complex, novel techniques are required for monitoring the diversity of the B cell immune response. Since antibodies recognize both linear and conformational protein and glycoprotein epitopes, recent advances in proteomic and glycomic technologies for rapid display of antigenic structures are leading to methods for proteome-wide immune monitoring. Here, we review different approaches for protein display for immune monitoring, and provide methods for in situ protein display for the rapid detection and validation of antibody repertoires.


Tuning the fluorescence of calcium-discharged photoprotein obelin via mutating at the His22-Phe88-Trp92 triad - a QM/MM study.
The fluorescence (FL) of calcium-discharged photoprotein (CaDP) can be altered by easily mutating CaDP without modifying coelenteramide (CLM), which is the decarboxylation product of coelenterazine in calcium-regulated photoprotein. The His22-Phe88-Trp92 triad (the ordering numbers of three amino acids are sorted by a crystal structure (PDB: 2F8P) of calcium-discharged obelin, i.e., CaDP-obelin) is closely related to CaDP-obelin FL, since it exists in close proximity to the 5-p-hydroxyphenyl of CLM. Therefore, it is important to thoroughly investigate how the mutations of this triad affect the emission color of CaDP-obelin FL. In this study, by mutating wild-type CaDP-obelin (WT) at the His22-Phe88-Trp92 triad, we theoretically constructed its nine mutants of separable FL colors. Through combined quantum mechanics and molecular mechanics (QM/MM) calculations and molecular dynamics (MD) simulations, the influence of the mutations of this triad on the CaDP-obelin FL was analyzed considering the H-bond effect and the charge effect. This study demonstrated that the mutations at the His22-Phe88-Trp92 triad redistribute the charges on the D-π-A molecule, CLM, change the charge transfer from the D to the (π + A) moiety, and thereby alter the FL emission. Appending more negative charges on the phenolate moiety of CLM benefits the FL redshift.


The PqsE Active Site as a Target for Small Molecule Antimicrobial Agents against Pseudomonas aeruginosa.
The opportunistic pathogen Pseudomonas aeruginosa causes antibiotic-resistant, nosocomial infections in immuno-compromised individuals and is a high priority for antimicrobial development. Key to pathogenicity in P. aeruginosa are biofilm formation and virulence factor production. Both traits are controlled by the cell-to-cell communication process called quorum sensing (QS). QS involves the synthesis, release, and population-wide detection of signal molecules called autoinducers. We previously reported that the activity of the RhlR QS transcription factor depends on a protein-protein interaction with the hydrolase, PqsE, and PqsE catalytic activity is dispensable for this interaction. Nonetheless, the PqsE-RhlR interaction could be disrupted by the substitution of an active site glutamate residue with tryptophan [PqsE(E182W)]. Here, we show that disruption of the PqsE-RhlR interaction via either the E182W change or alteration of PqsE surface residues that are essential for the interaction with RhlR attenuates P. aeruginosa infection in a murine host. We use crystallography to characterize the conformational changes induced by the PqsE(E182W) substitution to define the mechanism underlying disruption of the PqsE-RhlR interaction. A loop rearrangement that repositions the E280 residue in PqsE(E182W) is responsible for the loss of interaction. We verify the implications garnered from the PqsE(E182W) structure using mutagenic, biochemical, and additional structural analyses. We present the next generation of molecules targeting the PqsE active site, including a structure of the tightest binding of these compounds, BB584, in complex with PqsE. The findings presented here provide insights into drug discovery against P. aeruginosa with PqsE as the target.


Application of a diagnostic algorithm for inherited thrombocytopenias to 46 consecutive patients.
The Italian Gruppo di Studio delle Piastrine recently developed a diagnostic algorithm to assist clinicians in the diagnosis of inherited thrombocytopenias. This algorithm is based on the simplest possible diagnostic investigations and can also be used in centers that are not highly specialized. The aim of the present study was to validate this diagnostic algorithm by applying it to a case series of genetic thrombocytopenias. The diagnostic algorithm was applied retrospectively to 46 consecutive patients observed during the last five years at a single institution. Twenty-eight were affected by defined illnesses or their variants, while 18 had a disorder that did not fit the criteria for any known genetic thrombocytopenia. The study was based on the evaluation of clinical records and laboratory tests. The diagnostic algorithm recognized: 4 homozygous and 4 heterozygous Bernard-Soulier syndromes, 11 MYH9-related diseases, one von WillebrandOs disease type 2B, one gray platelet syndrome and one X-linked thrombocytopenia with thalassemia. Moreover, it identified 4 patients with the clinical and laboratory features of heterozygous Bernard-Soulier syndrome not caused by mutations in the coding region of the GPIbalpha, GPIbbeta, GPIX or GPV genes, and two patients with the clinical phenotype of MYH9-related disease but without MYH9 mutations. Since the diagnostic flow chart did not allow prompt recognition of two subjects with MYH9-related disease, we introduced a small change to the previously proposed flow chart to obviate this defect. The diagnostic algorithm correctly diagnosed 26 of 28 patients with known disorders or phenotypic variants of known disorders. By a simple modification of the investigation sequence, its sensitivity reached 100%. The algorithm also identified 18 patients with new, as yet uncharacterized forms of genetic thrombocytopenia.


Effects of increased CO2 and temperature on the physiological characteristics of the golden tide blooming macroalgae Sargassum horneri in the Yellow Sea, China.
The golden tide, caused by the brown algae Sargassum horneri, exerts severe influences on the Pyropia aquaculture of Jiangsu coast, China. To study the outbreak of the golden tide in response to increasing greenhouse gas emissions, S. horneri was cultured under four conditions: ambient condition (10 °C, 400 μatm), elevated temperature condition (14 °C, 400 μatm), elevated CO2 level (10 °C, 1000 μatm), and potential greenhouse condition (14 °C, 1000 μatm). The growth, photosynthetic performances, and inorganic carbon affinity of S. horneri were studied. The results showed that elevated temperature exerted a more pronounced positive influence on S. horneri growth, photosynthesis, and carbon assimilation than CO2 enrichment. The growth of S. horneri was significantly improved by moderately elevated temperatures, especially under concurrently elevated CO2 levels. This suggests that the greenhouse effect will benefit growth and carbon sequestration of S. horneri, which may enhance the frequency and scale of golden tides.


Size-dependent phase diagrams of metallic alloys: A Monte Carlo simulation study on order-disorder transitions in Pt-Rh nanoparticles.
Nanoparticles of Pt-Rh were studied by means of lattice-based Monte Carlo simulations with respect to the stability of ordered D0(22)- and 40-phases as a function of particle size and composition. By thermodynamic integration in the semi-grand canonical ensemble, phase diagrams for particles with a diameter of 7.8 nm, 4.3 nm and 3.1 nm were obtained. Size-dependent trends such as the lowering of the critical ordering temperature, the broadening of the compositional stability range of the ordered phases, and the narrowing of the two-phase regions were observed and discussed in the context of complete size-dependent nanoparticle phase diagrams. In addition, an ordered surface phase emerges at low temperatures and low platinum concentration. A decrease of platinum surface segregation with increasing global platinum concentration was observed, when a second, ordered phase is formed inside the core of the particle. The order-disorder transitions were analyzed in terms of the Warren-Cowley short-range order parameters. Concentration-averaged short-range order parameters were used to remove the surface segregation bias of the conventional short-range order parameters. Using this procedure, it was shown that the short-range order in the particles at high temperatures is bulk-like.


Identifying Components in 3D Density Maps of Protein Nanomachines by Multi-scale Segmentation.
Segmentation of density maps obtained using cryo-electron microscopy (cryo-EM) is a challenging task, and is typically accomplished by time-intensive interactive methods. The goal of segmentation is to identify the regions inside the density map that correspond to individual components. We present a multi-scale segmentation method for accomplishing this task that requires very little user interaction. The method uses the concept of scale space, which is created by convolution of the input density map with a Gaussian filter. The latter process smoothes the density map. The standard deviation of the Gaussian filter is varied, with smaller values corresponding to finer scales and larger values to coarser scales. Each of the maps at different scales is segmented using the watershed method, which is very efficient, completely automatic, and does not require the specification of seed points. Some detail is lost in the smoothing process. A sharpening process reintroduces detail into the segmentation at the coarsest scale by using the segmentations at the finer scales. We apply the method to simulated density maps, where the exact segmentation (or ground truth) is known, and rigorously evaluate the accuracy of the resulting segmentations.


Effects of hyperglycemia on variability of RR, QT and corrected QT intervals in Type 1 diabetic patients.
In this study, we evaluated the effects of hyperglycemia on the variability of RR (HRV), QT interval variability (QTV) and corrected QT interval variability (QTcV) during hyperglycemic and non-hyperglycemic conditions in six Type 1 diabetic patients at nights. The aim of this study was to investigate the association of high blood glucose levels with autonomic modulation of heart rate and variation in ventricular repolarization. Blood glucose level (BGL) threshold for defining hyperglycemia state was set at 8.33 mmol/l. Variability of RR, QT and corrected QT intervals during hyperglycemic and non-hyperglycemic were quantified using time and frequency domain measures. Hypomon® device was used to monitor ECG signals and acquire RR and QT intervals in Type 1 diabetic patients overnight. The results indicated that time and frequency domain HRV variables were significantly decreased under hyperglycemic condition and inversely correlated with BGL. QTV parameters also reduced when BGL increased and time domain measures of QTV were inversely associated with BGL. Variability in QTc interval was much less than in the QT interval and demonstrated a lower SDNN and LF power. We concluded that certain components of HRV, time-domain measures of QTV and QTc but not QTcV are strongly correlated to high blood glucose levels and can be good markers to identify hyperglycemic events in T1DM.


Adeno-Associated Viral Vectors Transduce Mature Human Adipocytes in Three-Dimensional Slice Cultures.
Adipose tissue plays a pivotal role, both in the regulation of energy homeostasis and as an endocrine organ. Consequently, adipose tissue dysfunction is closely related to insulin resistance, morbid obesity, and metabolic syndrome. To study molecular mechanisms and to develop novel therapeutic strategies, techniques are required to genetically modify mature adipocytes. Here, we report on adeno-associated viral (AAV) vectors as a versatile tool to transduce human mature adipocytes in organotypic three-dimensional tissue cultures.


Epithelial-Mesenchymal Transitions during Neural Crest and Somite Development.
Epithelial-to-mesenchymal transition (EMT) is a central process during embryonic development that affects selected progenitor cells of all three germ layers. In addition to driving the onset of cellular migrations and subsequent tissue morphogenesis, the dynamic conversions of epithelium into mesenchyme and vice-versa are intimately associated with the segregation of homogeneous precursors into distinct fates. The neural crest and somites, progenitors of the peripheral nervous system and of skeletal tissues, respectively, beautifully illustrate the significance of EMT to the above processes. Ongoing studies progressively elucidate the gene networks underlying EMT in each system, highlighting the similarities and differences between them. Knowledge of the mechanistic logic of this normal ontogenetic process should provide important insights to the understanding of pathological conditions such as cancer metastasis, which shares some common molecular themes.


Factors associated with intern fatigue.
Prior data suggest that fatigue adversely affects patient safety and resident well-being. ACGME duty hour limitations were intended, in part, to reduce resident fatigue, but the factors that affect intern fatigue are unknown. To identify factors associated with intern fatigue following implementation of duty hour limitations. Cross-sectional confidential survey of validated questions related to fatigue, sleep, and stress, as well as author-developed teamwork questions. Interns in cognitive specialties at the University of California, San Francisco. Univariate statistics characterized the distribution of responses. Pearson correlations elucidated bivariate relationships between fatigue and other variables. Multivariate linear regression models identified factors independently associated with fatigue, sleep, and stress. Of 111 eligible interns, 66 responded (59%). In a regression analysis including gender, hours worked in the previous week, sleep quality, perceived stress, and teamwork, only poorer quality of sleep and greater perceived stress were significantly associated with fatigue (p < 0.001 and p = 0.02, respectively). To identify factors that may affect sleep, specifically duty hours and stress, a secondary model was constructed. Only greater perceived stress was significantly associated with diminished sleep quality (p = 0.04), and only poorer teamwork was significantly associated with perceived stress (p < 0.001). Working >80 h was not significantly associated with perceived stress, quality of sleep, or fatigue. Simply decreasing the number of duty hours may be insufficient to reduce intern fatigue. Residency programs may need to incorporate programmatic changes to reduce stress, improve sleep quality, and foster teamwork in order to decrease intern fatigue and its deleterious consequences.


Bitot's spots responsive and nonresponsive to vitamin A. Clinicopathologic correlations.
Conjunctival biopsy specimens from patients with Bitot's spot responsive and nonresponsive to vitamin A were studied by light and electron microscopy. In both types, the lesions demonstrated keratinization with granular cells, irregular maturation, inflammatory infiltration of the conjunctival substantia propria, and loss of goblet cells. Only in the responsive cases were these changes generalized. Prominent Bitot's spots represented massive accumulations of Gram-positive bacilli and keratin debris. Responsive cases improved histologically within seven days of treatment, and goblet cells began to return within two weeks. These results support our previous suggestions that there is little basis for attempting to differentiate, clinically, between the two types of lesions and that at least some nonresponsive lesions represent a persistent metaplastic change induced during a prior episode of vitamin A deficiency.


How do we raise media bias awareness effectively? Effects of visualizations to communicate bias.
Media bias has a substantial impact on individual and collective perception of news. Effective communication that may counteract its potential negative effects still needs to be developed. In this article, we analyze how to facilitate the detection of media bias with visual and textual aids in the form of (a) a forewarning message, (b) text annotations, and (c) political classifiers. In an online experiment, we randomized 985 participants to receive a biased liberal or conservative news article in any combination of the three aids. Meanwhile, their subjective perception of media bias in this article, attitude change, and political ideology were assessed. Both the forewarning message and the annotations increased media bias awareness, whereas the political classification showed no effect. Incongruence between an articles' political position and individual political orientation also increased media bias awareness. Visual aids did not mitigate this effect. Likewise, attitudes remained unaltered.


Factors Associated With Response to Biofeedback Therapy for Dyssynergic Defecation.
Biofeedback therapy is effective for dyssynergic defecation (DD), but it is not widely available or reimbursed, and is labor intensive. It is therefore important to select the appropriate patients for this treatment. We investigated symptoms and demographic, manometric, and other factors associated with outcomes of biofeedback therapy in patients with DD. We performed a post hoc analysis of 2 prospective studies of biofeedback therapy in 127 adult outpatients (18-75 years old, 120 female) with chronic constipation who failed to respond to treatment with dietary fiber or laxatives (>1 year) and were diagnosed with DD based on standard criteria. In each study, patients received 1-hour, biweekly office biofeedback therapy (6 sessions) or home biofeedback therapy with a device. A therapist used visual feedback, postural, and diaphragmatic breathing techniques to teach subjects to improve defecation. Treatment success was defined by a composite of normalization of dyssynergia pattern and increase of 20 mm in baseline bowel satisfaction score. Factors were compared between the treatment success and failure groups. Intention-to-treat analysis was performed. Of the 127 patients enrolled, 77 (61%) had treatment success. Dyssynergia was corrected in 78% of patients and bowel satisfaction improved in 64% of patients. Baseline demographic features, constipation symptoms, manometric and sensory parameters, balloon expulsion time, and colonic transit results were similar between treatment failure and success groups. Patients with lower baseline bowel satisfaction score (P = .008) and patients who used digital maneuvers (P = .04) were more likely to have successful biofeedback therapy. Biofeedback therapy is successful in more than 60% of patients with DD. Patients who used digital maneuvers and patients with lower baseline levels of bowel satisfaction were more likely to have treatment success, whereas other factors were not associated with success. Biofeedback therapy should be offered to all patients with DD, irrespective of baseline symptoms or anorectal physiology findings.