Toward the Darwinian transition: Switching between distributed and speciated states in a simple model of early life.
It has been hypothesized that in the era just before the last universal common ancestor emerged, life on earth was fundamentally collective. Ancient life forms shared their genetic material freely through massive horizontal gene transfer (HGT). At a certain point, however, life made a transition to the modern era of individuality and vertical descent. Here we present a minimal model for stochastic processes potentially contributing to this hypothesized "Darwinian transition." The model suggests that HGT-dominated dynamics may have been intermittently interrupted by selection-driven processes during which genotypes became fitter and decreased their inclination toward HGT. Stochastic switching in the population dynamics with three-point (hypernetwork) interactions may have destabilized the HGT-dominated collective state and essentially contributed to the emergence of vertical descent and the first well-defined species in early evolution. A systematic nonlinear analysis of the stochastic model dynamics covering key features of evolutionary processes (such as selection, mutation, drift and HGT) supports this view. Our findings thus suggest a viable direction out of early collective evolution, potentially enabling the start of individuality and vertical Darwinian evolution.


Cortical spreading depolarisation-induced facial hyperalgesia, photophobia and hypomotility are ameliorated by sumatriptan and olcegepant.
Cortical spreading depolarisation (CSD), the neural mechanism underlying migraine aura, may cause headache by sensitising the trigeminal system. Photophobia, the most bothersome accompanying symptom during migraine attacks, is more prevalent in migraine with aura than in migraine without aura. Whether CSD plays a role in developing photophobia remains unknown. Moreover, migraine-induced physical hypoactivity contributes to loss of productivity. We aimed to investigate the development of trigeminal sensitisation, photophobia and locomotive abnormality after KCl-induced CSD using 86 male C57BL/6 mice. Sham-operated mice were used as controls. We confirmed the presence of trigeminal sensitisation and photophobia at 24 h after CSD. CSD-subjected mice also exhibited significantly reduced locomotive activity in both light and dark zones. Hence, the CSD-induced hypomobility was likely to be independent of photophobia. The 5-HT1B/1D agonist, sumatriptan, corrected all these CSD-induced abnormalities. Moreover, dose dependency was demonstrated in the ameliorating effect of the calcitonin gene-related peptide (CGRP) receptor antagonist, olcegepant, on these abnormalities. Sumatriptan and olcegepant improved mouse locomotion with therapeutic lags ranging from 20 to 30 min. Collectively, CSD caused trigeminal sensitisation, photophobia and hypomobility that persisted for at least 24 h by a mechanism involving the 5-HT1B/1D and CGRP activity.


Oxidative cleavage of the pentyl side-chain of cannabinoids. Identification of new biotransformation pathways in the metabolism of 4'-hydroxy-delta-9-tetrahydrocannabinol in the mouse.
During an investigation of the mechanisms leading to the formation of metabolites of cannabinoids in which the pentyl side chain is reduced to 2, 3 or 4 carbon atoms, the further metabolism of 4'-hydroxy-delta 9-tetrahydrocannabinol was investigated in vivo in mice. Metabolites were extracted with ethyl acetate, concentrated by chromatography on Sephadex LH-20 and identified by GC/MS. Ten metabolites were identified and a further two had tentative structural assignments made. The major metabolic route, in common with that seen with most cannabinoids, was hydroxylation at the allylic 11-position, followed by oxidation to a carboxylic acid. Additional hydroxylation occurred at C-8. Abundant metabolites were also formed by oxidative cleavage of the pentyl side chain. The major metabolites of this type had lost the terminal two carbon atoms to give compounds containing a carboxyethyl side chain. This is the major product normally produced by beta-oxidation of the acid formed from 5'-hydroxy-delta 9-tetrahydrocannabinol. Trace concentrations of two other acids that appeared to have a carboxypropyl side chain were also found. The results show that, in addition to beta-oxidation, initiated by hydroxylation at the 5'-carbon atom (omega-hydroxylation), at least one other oxidative route, initiated by omega-1-hydroxylation, is involved in the production of metabolites with two carbon atoms missing from the pentyl side chain. This pathway does not seem to have been characterized as a biotransformation mechanism in drug metabolism and a possible mechanism is suggested.


A comparison of upper and lower body energetics during high-intensity exercise.
The aim of the present study was to measure and compare the aerobic, anaerobic alactic and anaerobic lactic energy system contribution during the 30-sec Wingate anaerobic test (WAnT) for the upper and lower body within the same individuals. Physically active men (N.=14) completed two WAnTs on an electronic arm ergometer and a cycle ergometer separated by three days. A fly wheel braking force corresponding to 5% and 7.5% of the participants body weight was used for the upper and lower body WAnTs respectively. Oxygen uptake and blood lactate were measured before, during and after both WAnTs, and body composition was measured using dual-energy X-ray absorptiometry (DEXA). The anaerobic lactic energy system contribution was significantly (P<0.01) higher during the upper body (60.3±5.6%) compared to the lower body (46.9±6.9%) WAnT. The contribution of the anaerobic alactic system was significantly higher (P<0.01) during the lower body (36.5±6.3%) compared to the upper body (28.3±4.9%) WAnT, with the aerobic system contribution significantly (P<0.05) higher for the lower body (16.8±2.5%) compared to the upper body (11.4±1.4%) WAnT. The anaerobic lactic energy system provides over 60% of the energy requirements during an upper body WAnT but provides less the 50% during the lower body WAnT. In contrast, the aerobic and anaerobic alactic energy system contribution was significantly less for the upper body WAnT compared to the lower body WAnT.


Intense pulsed light treatment of chronic mid-body Achilles tendinopathy: A double blind randomised placebo-controlled trial.
We conducted a randomised controlled trial to determine whether active intense pulsed light (IPL) is an effective treatment for patients with chronic mid-body Achilles tendinopathy. A total of 47 patients were randomly assigned to three weekly therapeutic or placebo IPL treatments. The primary outcome measure was the Victorian Institute of Sport Assessment - Achilles (VISA-A) score. Secondary outcomes were a visual analogue scale for pain (VAS) and the Lower Extremity Functional Scale (LEFS). Outcomes were recorded at baseline, six weeks and 12 weeks following treatment. Ultrasound assessment of the thickness of the tendon and neovascularisation were also recorded before and after treatment. There was no significant difference between the groups for any of the outcome scores or ultrasound measurements by 12 weeks, showing no measurable benefit from treatment with IPL in patients with Achilles tendinopathy.


Nucleotide-dependent self-assembly of Nucleoside Diphosphate Kinase (NDPK) in vitro.
In addition to their role in nucleotide homeostasis, members of the Nucleoside Diphosphate Kinase (NDPK) family have been implicated in tumor metastasis, cell migration and vesicle trafficking. Although its role in most cases depends on nucleotide catalysis, a precise understanding of how the catalytic activity of NDPK supports its function in diverse processes is lacking. Here we report that wild type, but not catalytically inactive (H118C) NDPKB, undergoes dynamic self-assembly into ordered 20-25 nm diameter filaments in vitro. Self-assembly is nucleoside triphosphate dependent, GTP being most effective at promoting polymer formation. In addition, polymerization appears to depend on formation of the phosphoryl-Histidine intermediate of the enzyme, suggesting a previously unappreciated conformational change in NDPK during its catalytic cycle. We hypothesize that the observed nucleotide-dependent self-assembly property of NDPKB may reflect a key feature of NDPK enzymes that enables their function in diverse processes.


SMART, a simple modular architecture research tool: identification of signaling domains.
Accurate multiple alignments of 86 domains that occur in signaling proteins have been constructed and used to provide a Web-based tool (SMART: simple modular architecture research tool) that allows rapid identification and annotation of signaling domain sequences. The majority of signaling proteins are multidomain in character with a considerable variety of domain combinations known. Comparison with established databases showed that 25% of our domain set could not be deduced from SwissProt and 41% could not be annotated by Pfam. SMART is able to determine the modular architectures of single sequences or genomes; application to the entire yeast genome revealed that at least 6.7% of its genes contain one or more signaling domains, approximately 350 greater than previously annotated. The process of constructing SMART predicted (i) novel domain homologues in unexpected locations such as band 4.1-homologous domains in focal adhesion kinases; (ii) previously unknown domain families, including a citron-homology domain; (iii) putative functions of domain families after identification of additional family members, for example, a ubiquitin-binding role for ubiquitin-associated domains (UBA); (iv) cellular roles for proteins, such predicted DEATH domains in netrin receptors further implicating these molecules in axonal guidance; (v) signaling domains in known disease genes such as SPRY domains in both marenostrin/pyrin and Midline 1; (vi) domains in unexpected phylogenetic contexts such as diacylglycerol kinase homologues in yeast and bacteria; and (vii) likely protein misclassifications exemplified by a predicted pleckstrin homology domain in a Candida albicans protein, previously described as an integrin.


Power calculations for preclinical studies using a K-sample rank test and the Lehmann alternative hypothesis.
Power calculations in a small sample comparative study, with a continuous outcome measure, are typically undertaken using the asymptotic distribution of the test statistic. When the sample size is small, this asymptotic result can be a poor approximation. An alternative approach, using a rank based test statistic, is an exact power calculation. When the number of groups is greater than two, the number of calculations required to perform an exact power calculation is prohibitive. To reduce the computational burden, a Monte Carlo resampling procedure is used to approximate the exact power function of a k-sample rank test statistic under the family of Lehmann alternative hypotheses. The motivating example for this approach is the design of animal studies, where the number of animals per group is typically small.


Integration versus apartheid in post-Roman Britain: a response to Thomas et Al. (2008).
The genetic surveys of the population of Britain conducted by Weale et al. and Capelli et al. produced estimates of the Germani immigration into Britain during the early Anglo-Saxon period, c.430-c.730. These estimates are considerably higher than the estimates of archaeologists. A possible explanation suggests that an apartheid-like social system existed in the early Anglo-Saxon kingdoms resulting in the Germani breeding more quickly than the Britons. Thomas et al. attempted to model this suggestion and showed that it was a possible explanation if all Anglo-Saxon kingdoms had such a system for up to 400 years. I noted that their explanation ignored the probability that Germani have been arriving in Britain for at least the past three millennia, including Belgae and Roman soldiers, and not only during the early Anglo-Saxon period. I produced a population model for Britain taking into account this long term, low level migration that showed that the estimates could be reconciled without the need for introducing an apartheid-like system. In turn, Thomas et al. responded, criticizing my model and arguments, which they considered persuasively written but wanting in terms of methodology, data sources, underlying assumptions, and application. Here, I respond in detail to those criticisms and argue that it is still unnecessary to introduce an apartheid-like system in order to reconcile the different estimates of Germani arrivals. A point of confusion is that geneticists are interested in ancestry, while archaeologists are interested in ethnicity: it is the bones, not the burial rites, which are important in the present context.


Whole-exome sequencing of muscle-invasive bladder cancer identifies recurrent mutations of UNC5C and prognostic importance of DNA repair gene mutations on survival.
Because of suboptimal outcomes in muscle-invasive bladder cancer even with multimodality therapy, determination of potential genetic drivers offers the possibility of improving therapeutic approaches and discovering novel prognostic indicators. Using pTN staging, we case-matched 81 patients with resected ≥pT2 bladder cancers for whom perioperative chemotherapy use and disease recurrence status were known. Whole-exome sequencing was conducted in 43 cases to identify recurrent somatic mutations and targeted sequencing of 10 genes selected from the initial screening in an additional 38 cases was completed. Mutational profiles along with clinicopathologic information were correlated with recurrence-free survival (RFS) in the patients. We identified recurrent novel somatic mutations in the gene UNC5C (9.9%), in addition to TP53 (40.7%), KDM6A (21.0%), and TSC1 (12.3%). Patients who were carriers of somatic mutations in DNA repair genes (one or more of ATM, ERCC2, FANCD2, PALB2, BRCA1, or BRCA2) had a higher overall number of somatic mutations (P = 0.011). Importantly, after a median follow-up of 40.4 months, carriers of somatic mutations (n = 25) in any of these six DNA repair genes had significantly enhanced RFS compared with noncarriers [median, 32.4 vs. 14.8 months; hazard ratio of 0.46, 95% confidence interval (CI), 0.22-0.98; P = 0.0435], after adjustment for pathologic pTN staging and independent of adjuvant chemotherapy usage. Better prognostic outcomes of individuals carrying somatic mutations in DNA repair genes suggest these mutations as favorable prognostic events in muscle-invasive bladder cancer. Additional mechanistic investigation into the previously undiscovered role of UNC5C in bladder cancer is warranted.


Changes in retinal and choroidal gene expression during development of refractive errors in chicks.
During growth, the retina analyzes the projected image to achieve a close match between eye length and focal length. Because the messengers released by retina and choroid are largely unknown, genes that are differently expressed in response to changes in the retinal image were identified. In addition, because glucagon may be important in the visual control of eye growth, the transcript levels of proglucagon were studied. Reverse transcription-polymerase chain reaction differential display was used to identify genes that were differentially expressed in chick eyes that were deprived of sharp vision or treated with positive or negative lenses. Differences were analyzed through sequencing and database searches and confirmed by Northern blot analyses. Combining 40 and 33 arbitrary primers with 3 oligo-dT-primers, approximately 48% and 40% of the retinal and choroidal mRNAs were screened, respectively. Twelve differences were detected in retinal tissue and five in choroidal tissue after 6 to 24 hours of exposure to defocus. Only one of 10 sequenced products could be identified as cytochrome-c oxidase, subunit I. Northern blot analysis confirmed its twofold upregulation after positive lens wear and also changes in four other unknown genes. Finally, it was shown that retinal glucagon mRNA content increased after treatment with positive lenses. Visual conditions that induce refractive errors produce changes in gene expression in retina and choroid within 1 day. In line with previous immunohistochemical data, it was found that the amount of glucagon mRNA was upregulated during wearing of positive lenses.


On the thermodynamics of folding of an i-motif DNA in solution under favorable conditions.
Under slightly acidic conditions, cytosine-rich DNA sequences can form non-canonical secondary structures called i-motifs, which occur as four stretches of cytosine repeats form hemi-protonated C·C+ base pairs. The growing interest in the i-motif structures as important components in functional DNA-based nanotechnology or as potential targets of anticancer drugs, increases the need for a deep understanding of the energetics of their structural transitions. Here, a combination of spectroscopic and calorimetric techniques is used to unravel the thermodynamics of folding of an i-motif DNA under favorable conditions. The results give new insights into the energetic aspects of i-motifs and show that thermodynamic and thermal stability are related but not identical properties of such DNA structures.


Management of elderly patients with sustained hypertension.
To assess the clinical benefits of treating hypertension in elderly patients and to derive practical guidelines regarding indications, goals, and forms of treatment. Review of six published randomised trials. Active treatment of hypertension in elderly patients was associated with significant improvements in several indices of cardiovascular morbidity and mortality, particularly the incidence of fatal and non-fatal strokes. On the basis of the trial data, combined systolic and diastolic hypertension was defined as a sustained systolic pressure greater than 160 mmHg and diastolic pressure greater than 90 mmHg. There is convincing evidence that efforts should be made to reduce both systolic and diastolic pressures to below these levels in patients up to the age of 80 years. Isolated systolic hypertension was defined as a systolic pressure greater than 160 mmHg in the presence of a diastolic pressure less than 90 mmHg. Two trials reported benefit from the treatment of isolated systolic hypertension in patients up to the age of 80, and further trials are underway to support or refute this recommendation. Diuretics have an established role in the management of hypertension in elderly patients; beta adrenoceptor antagonists have given variable results, and the benefits are less impressive than with diuretic based regimens. Newer agents show promise in the treatment of elderly patients, particularly in the presence of coexisting disease, but their effects on morbidity and mortality have not been evaluated in large randomised trials. Diuretics rather than beta blockers are the treatment of choice for patients with uncomplicated hypertension, but combinations of drugs may be required in as many as 50% of patients.


Deep EHR Spotlight: a Framework and Mechanism to Highlight Events in Electronic Health Records for Explainable Predictions.
The wide adoption of Electronic Health Records (EHR) has resulted in large amounts of clinical data becoming available, which promises to support service delivery and advance clinical and informatics research. Deep learning techniques have demonstrated performance in predictive analytic tasks using EHRs yet they typically lack model result transparency or explainability functionalities and require cumbersome pre-processing tasks. Moreover, EHRs contain heterogeneous and multi-modal data points such as text, numbers and time series which further hinder visualisation and interpretability. This paper proposes a deep learning framework to: 1) encode patient pathways from EHRs into images, 2) highlight important events within pathway images, and 3) enable more complex predictions with additional intelligibility. The proposed method relies on a deep attention mechanism for visualisation of the predictions and allows predicting multiple sequential outcomes.


Stem Lagomorpha and the antiquity of Glires.
We describe several fossils referable to Gomphos elkema from deposits close to the Paleocene-Eocene boundary at Tsagan Khushu, Mongolia. Gomphos shares a suite of cranioskeletal characters with extant rabbits, hares, and pikas but retains a primitive dentition and jaw compared to its modern relatives. Phylogenetic analysis supports the position of Gomphos as a stem lagomorph and excludes Cretaceous taxa from the crown radiation of placental mammals. Our results support the hypothesis that rodents and lagomorphs radiated during the Cenozoic and diverged from other placental mammals close to the Cretaceous-Tertiary boundary.


Targeting the NG2/CSPG4 proteoglycan retards tumour growth and angiogenesis in preclinical models of GBM and melanoma.
Aberrant expression of the progenitor marker Neuron-glia 2 (NG2/CSPG4) or melanoma proteoglycan on cancer cells and angiogenic vasculature is associated with an aggressive disease course in several malignancies including glioblastoma multiforme (GBM) and melanoma. Thus, we investigated the mechanism of NG2 mediated malignant progression and its potential as a therapeutic target in clinically relevant GBM and melanoma animal models. Xenografting NG2 overexpressing GBM cell lines resulted in increased growth rate, angiogenesis and vascular permeability compared to control, NG2 negative tumours. The effect of abrogating NG2 function was investigated after intracerebral delivery of lentivirally encoded shRNAs targeting NG2 in patient GBM xenografts as well as in established subcutaneous A375 melanoma tumours. NG2 knockdown reduced melanoma proliferation and increased apoptosis and necrosis. Targeting NG2 in two heterogeneous GBM xenografts significantly reduced tumour growth and oedema levels, angiogenesis and normalised vascular function. Vascular normalisation resulted in increased tumour invasion and decreased apoptosis and necrosis. We conclude that NG2 promotes tumour progression by multiple mechanisms and represents an amenable target for cancer molecular therapy.


The role of health literacy in patient-physician communication.
Patients' health literacy is increasingly recognized as a critical factor affecting patient-physician communication and health outcomes. We reviewed research on health literacy, examined its impact on patient-physician communication, and offer recommendations to enhance communication with patients who have poor health literacy. We conducted a literature review using the MEDLINE database for January 1966 through July 2001. The keywords "literacy" and "health literacy" were searched independently and in combination with the medical subject headings (MeSH) "physician-patient communication," "communication," and "reading." Poor health literacy is common, especially among elderly patients. More than 33% of patients ages 65 and older have inadequate or marginal health literacy, as do up to 80% of patients in public hospital settings. Patients with poor health literacy have a complex array of communication difficulties, which may affect health outcomes. Such patients report worse health status and have less understanding about their medical conditions and treatment; they may have increased hospitalization rates. Professional and public awareness of the health literacy issue must be increased, beginning with education of medical students and physicians and improved patient-physician communication skills. Future research needs to address identification of optimal methods for communicating with patients who have low literacy skills. This should focus on the effect of poor health literacy on patients' ability to communicate their history and physicians' ability to solicit information, as well as identifying the most-effective techniques to educate patients.


Evaluation of Lymphocyte Subgroups in Children With Down Syndrome.
In this study, lymphocyte subgroups including blood CD3, CD4, CD8, CD4/CD8, CD19, and CD16.56 values were analyzed in children with Down syndrome (DS). The study includes 85 children with DS, followed at Department of Pediatrics, Faculty of Medicine, Yüzüncü Yil University and 64 healthy age-matched control participants. Blood CD3, CD4, CD8, CD4/CD8, CD19, and CD16.56 values were examined in both the groups. Significantly decreased blood CD3, CD4, and CD19 values were found in the study group (P < .05) when compared with the control group. In conclusion, we would like to emphasize that blood CD3, CD4, and CD19 levels were found to be decreased in children with DS. Based on these finding, we think that these decreased lymphocyte subgroups might be responsible for increased susceptibility to infections in children with DS.


Health as a human right: an epidemiologist's perspective on the public health.
The modern idea of health as a human right is examined, as it evolved from the 18th century, in terms of its origins, its essential content, and 20th-century attempts at implementation. Equity for social groups is seen as a guiding principle. Two attempts at implementation, in Britain and in Cuba, are examined for their effects on equity in health service and in health states. The British National Health Service achieved equity between social classes in services but failed to achieve it in health states (as measured by mortality). Deficiencies in commitment to public health services, it is argued, contributed largely to this failure. The Cuban experiment appears to have moved beyond equity solely in services and toward equity in health states. This success reflects an overall Cuban commitment to the public health. Two important elements of that commitment are, first, continuous evaluation with flexible response and, second, community involvement.


Long-term exposure to acidification disrupts reproduction in a marine invertebrate.
Climate change research is advancing to more complex and more comprehensive studies that include long-term experiments, multiple life-history stages, multi-population, and multi-trait approaches. We used a population of the barnacle Balanus improvisus known to be sensitive to short-term acidification to determine its potential for long-term acclimation to acidification. We reared laboratory-bred individuals (as singles or pairs), and field-collected assemblages of barnacles, at pH 8.1 and 7.5 (≈ 400 and 1600 μatm pCO2 respectively) for up to 16 months. Acidification caused strong mortality and reduced growth rates. Acidification suppressed respiration rates and induced a higher feeding activity of barnacles after 6 months, but this suppression of respiration rate was absent after 15 months. Laboratory-bred barnacles developed mature gonads only when they were held in pairs, but nonetheless failed to produce fertilized embryos. Field-collected barnacles reared in the laboratory for 8 months at the same pH's developed mature gonads, but only those in pH 8.1 produced viable embryos and larvae. Because survivors of long-term acidification were not capable of reproducing, this demonstrates that B. improvisus can only partially acclimate to long-term acidification. This represents a clear and significant bottleneck in the ontogeny of this barnacle population that may limit its potential to persist in a future ocean.


Factors affecting mode of delivery in women of advanced maternal age.
With the implementation of the two-child policy in China, an increased number of women of advanced maternal age (AMA) have been giving birth. Formulating evidence-based guidance for the clinical management of this population is crucial. This retrospective study aimed to explore factors influencing the mode of delivery in women of AMA. Data on 350 women of AMA who delivered at Shanghai Putuo Maternity & Infant Health Hospital from January to June of 2016 were collected. Results indicated that most (114/134, 85%) of the multiparae chose delivery via cesarean section (CS) because of uterine scarring. There were significant differences in the body mass index (BMI) before pregnancy, BMI at delivery, gestational diabetes mellitus (GDM), pregnancy-induced hypertension (PIH), and placenta previa between the CS and vaginal delivery groups (P < 0.05 for all). The current results suggest that vaginal delivery is recommended for the first delivery whenever reasonable. Moreover, management of metabolic disorders during pregnancy is essential to effectively reduce the rate of CS among women of AMA.


Remote "hovering" with individuals with psychotic disorders and substance use: feasibility, engagement, and therapeutic alliance with a text-messaging mobile interventionist.
People with serious mental illnesses and substance abuse problems (i.e., dual diagnosis) constitute a particularly challenging and costly clinical group. This study evaluated the feasibility and acceptability of a novel model of care in which a mobile interventionist used mobile phone text messaging to remotely monitor and provide daily support to individuals with psychotic disorders and substance use. Seventeen participants with dual diagnosis were enrolled in a 12-week single-arm trial. A clinical social worker served as the mobile interventionist and sent daily text messages to participants' privately owned mobile phones to assess their medication adherence and clinical status. The mobile interventionist provided text-message feedback and support and suggested various coping strategies flexibly, in response to participants' replies to prompts. At the end of the trial, participants completed a usability and satisfaction measure and two self-rated measures of therapeutic alliance with their clinicians. In one version, participants rated their relationship with their mobile interventionist; in the second version, they rated their relationship with their community-based treatment team. Participants received an average of 139 text messages (SD = 37.5) each from the mobile interventionist over the 12-week trial. On average, participants responded to 87% of the mobile interventionist's messages that required a reply. More than 90% of participants thought the intervention was useful and rewarding and that it helped them be more effective and productive in their lives. Participants' assessments of their relationship with the mobile interventionist were positive. Paired-sample t-test found that the therapeutic alliance ratings participants provided for their mobile interventionist were significantly higher than those provided for their community-based treatment team clinicians, who they met with regularly. Our findings suggest that text-message "hovering" can be conducted successfully with individuals with psychotic disorders and substance abuse. Developing a cadre of mobile interventionists who are specifically trained on how to engage patients via mobile devices while adhering to ethical guidelines and regulatory standards may be an effective way to strengthen service delivery models, improve patient outcomes, and reduce costs.


Driving After Drinking Alcohol Associated with Insufficient Sleep and Insomnia among Student Athletes and Non-Athletes.
The proportion of university/college students (UCS) consuming alcohol is similar to the number of those reporting poor sleep, at approximately 30%, the proportion being greater in student athletes (SA). What remains to be understood is if poor sleep potentiates risky behaviors. Our aim was to examine the association among sleep difficulties, insomnia symptoms, and insufficient sleep on the risk of driving under the influence of alcohol in a sample of UCS and whether these associations were more pertinent in SA. Data from the National University/College Health Assessment was used from the years 2011⁻2014. Questions on number of drinks consumed and behaviors such as driving after drinking alcohol were related to answers to questions pertaining to sleep difficulties, insufficient sleep, and insomnia symptoms. Mean alcohol intake was of about 3 drinks; SA consumed significantly more than student non-athletes (SNA). Binge-drinking episodes were significantly higher among SA than SNA. Difficulty sleeping was associated with an increased likelihood of driving after any drinks and after 5 or more drinks in both groups, effects being stronger among SA. Insomnia was associated with an increased likelihood of driving after any drinks and after 5 or more drinks in SA and after 5 or more drinks in SNA. These effects were stronger among athletes. The present study found that self-reported difficulties sleeping, insomnia symptoms, and insufficient sleep are associated with driving after drinking alcohol. This relationship applied to driving after drinking any alcohol or binge drinking and was again stronger among SA than SNA.


Plasmodium coatneyi ring-infected erythrocyte surface antigens.
Plasmodium coatneyi produced ring-infected erythrocyte surface antigen (RESA) during infection of the rhesus monkey. This antigen was immunogenic and elicited an antibody response that was not persistent but was boosted by repeated infections in a manner similar to that seen in P. falciparum infections in humans. Preliminary data showed that the appearance and increasing titer of antibodies to P. coatneyi RESA-like antigen were associated with prolongation of intervals from inoculation to patency and with control of parasitemia. Studies using both immunofluorescence assay and Western blot analysis showed that P. coatneyi-immune rhesus serum cross-reacted with P. falciparum antigens, but P. falciparum immune human serum did not recognize P. coatneyi antigens in either assay. These results show that P. coatneyi expresses RESA-like antigen that elicits an antibody response similar to that observed for human antibody to P. falciparum RESA. However, antibodies to P. coatneyi did not cross-react with P. falciparum RESA in erythrocyte membrane immunofluorescence assay and dot immunoblot analysis, suggesting that different immunogenic epitopes are present on the two molecules. Our observations support the use of this primate model in RESA-based vaccine development.


Discovery of a diverse cave flora in China.
Few studies document plants in caves. Our field observations of a widespread and seemingly angiosperm-rich cave flora in SW China lead us to test the following hypotheses, 1) SW China caves contain a diverse vascular plant flora, 2) that this is a relic of a largely absent forest type lacking endemic species, and 3) that the light environment plants occupy in caves is not distinct from non-cave habitats. To do so we surveyed 61 caves and used species accumulation curves (SAC) to estimate the total diversity of this flora and used a subsample of 14 caves to characterise the light environment. We used regional floras and existing conservation assessments to evaluate the conservation value of this flora. We used observations on human disturbance within caves to evaluate anthropogenic activities. Four-hundred-and-eighteen vascular plant species were documented with SACs predicting a total diversity of 529-846. Ninety-three percent of the species documented are known karst forest species, 7% are endemic to caves and 81% of the species are angiosperms. We demonstrate that the light environment in caves is distinct to that of terrestrial habitats and that a subset of the flora likely grow in the lowest light levels documented for vascularised plants. Our results suggest that the proportion of species threatened with extinction is like that for the terrestrial habitat and that almost half of the entrance caverns sampled showed signs of human disturbance. We believe that this is the first time that such an extensive sample of cave flora has been undertaken and that such a diverse vascular plant flora has been observed in caves which we predict occurs elsewhere in SE Asia. We argue that the cave flora is an extension of the karst forest understory present prior to catastrophic deforestation in the 20thC. We suggest that within SW China caves serve as both refuges and a valuable source of germplasm for the restoration of karst forest. We also propose that caves represent a distinct habitat for plants that is most similar to that of the forest understory, but distinct with respect to the absence of trees, leaf litter, root mats, higher levels of atmospheric CO2, and lower diurnal and annual variation in temperature and humidity. We highlight tourism, agriculture and the absence of legislated protection of caves as the main current threats to this flora.


Status of insecticide susceptibility in Anopheles gambiae sensu lato and Anopheles funestus mosquitoes from western Kenya.
The status of resistance was investigated in Anopheles gambiae sensu lato and An. funestus (Diptera: Culicidae) mosquitoes from western Kenya to four classes of insecticides approved by World Health Organization for indoor residual spraying. The prevalence of the knockdown-resistance (kdr) mutation associated with resistance to pyrethroids and DDT was determined in An. gambiae s.l.. Standard World Health Organization diagnostic bioassay kits for DDT (an organochlorine), fenitrothion (an organophosphate), bendiocarb (a carbamate), and the pyrethoirds, lambdacyhalothrin and permethrin, were used. Knockdown every 10 min and mortality 24 h after exposure were noted. Controls not treated with insecticides and with the susceptible An. gambiae KISUMU strain were included in the bioassays. The presence of the kdr gene was determined using a standard diagnostic polymerase chain reaction assay. Over 98% mortality was observed for tests with all insecticides for both An. gambiae s.l. and An. funestus. Knockdown rates were not significantly different between An. gambiae s.l. and the KISUMU strain control. 50% and 95% knockdown times were either slightly lower than those for the KISUMU strain or higher by factors of less than 1.6. The mean frequency of the East African kdr mutation was 24.7% in An. gambiae sensu strictu. Based on conventional criteria where susceptibility is defined by mortality rates >98% 24 h after exposure, no evidence for resistance was found, implying that vector control measures employing any of the insecticides tested would be unhampered by resistance. The observed frequencies of the kdr mutation do not appear to compromise the effectiveness of the insecticides. The need for continuous monitoring of the status of insecticide resistance and of the impact of any observed resistance on the efficacy of vector control programs employing insecticides is apparent.


Prevalence, incidence, predictors and outcome of type 2 diabetes in Turkey.
To investigate prospectively the incidence, certain predictors and outcomes of type 2 diabetes (DM), as well as to determine its prevalence cross-sectionally, in a representative sample of Turkish men and women. Prospective evaluation of 3401 male and female participants (aged 48.2 +/-12 years). Follow-up constituted 19,050 person-years. Individuals with DM were diagnosed with criteria of the American Diabetes Association. Fatal and nonfatal coronary heart disease (CHD) was identified by clinical findings and Minnesota coding of resting electrocardiograms. Cut-points of > or = 95 cm in males and > or = 91 cm in females were selected for abdominal obesity. For prospective evaluations, cases with DM or CHD were excluded. Prevalence of DM in Turkish adults was estimated as 2.89 million (11.0% of the population aged > or = 35 years). Over a mean follow-up of 5.9 years, incident DM developed in 223 subjects, yielding an incidence per 1000 person-years of 11.0 in women and 12.4 in men. This corresponded to a 300,000 annual incidence. Following risk parameter levels but not HDL-cholesterol were significantly elevated at baseline in subjects developing DM compared to those without: age (5 years), waist girth (7 cm), blood pressure (12/6 mmHg), apolipoprotein B (7 mg/dl), total cholesterol (14 mg/dl), and fasting triglycerides (only in women, 52 mg/dl). Abdominal obesity (RR 2.61 [95%CI 1.87; 3.63]) and age in both genders, hypertension (RR 1.81 [95%CI 1.10; 2.98]) and low HDL-cholesterol in men alone were significant independent predictors of DM. Diabetes mellitus was a significant and independent predictor of fatal and nonfatal CHD, with a RR of 1.81 (95%CI 1.19; 2.75), after adjustment for sex, age, hypertension, waist circumference, serum total cholesterol and smoking status. The annual incidence of DM in Turkey rises very rapidly, currently stands at 300,000, and, hence, its prevalence also rises correspondingly. Insulin resistance appears to be a weak determinant of DM in Turkish women while abdominal obesity is the main determinant. Multivariately adjusted DM is a significant independent predictor of fatal and nonfatal CHD. These observations emphasize that measures to reverse or stop the "epidemic" of abdominal obesity are severely required.


Single-Case Experimental Design: Current Standards and Applications in Occupational Therapy.
Occupational therapy is a field with a long-standing history of recommending and implementing interventions designed to improve the quality of life of clients with disabilities. Often, the interventions are individualized to meet the needs of this diverse group of clients in dynamic settings. Identifying effective and efficient interventions for such a diverse group of clients and settings requires a flexible research approach. Single-case experimental designs (SCEDs) allow practitioners and researchers to answer experimental questions within the context of rigorous research designs. The purpose of this article is to highlight the similarities between the mission of occupational therapy and SCEDs. Recommendations for designing single-case studies with the framework provided by the Single-Case Reporting Guideline in Behavioral Interventions are provided. In addition, common problems and proposed solutions, along with implications for practitioners and researchers, are provided.


Pulmonary infection with Nocardia species: a report of 10 cases and review.
Pulmonary nocardiosis (PN) is an infrequent and severe infection due to Nocardia spp., microorganisms that may behave both as opportunists and as primary pathogens. The aim of this study and review was to evaluate the clinical features, evolution and prognostic factors of PN. The study group comprised 10 consecutive patients with pulmonary nocardiosis acquired in a community setting, diagnosed and followed in a tertiary teaching hospital. Chronic obstructive pulmonary disease (COPD), neoplastic disease and human immunodeficiency virus (HIV) infection were the most frequent predisposing factors. Four patients were receiving corticosteroid treatment. Clinical course was chronic and diagnosis was delayed 3 weeks or more in seven of the patients. Lobar or multilobar condensation was the most frequent radiographic pattern. Antimicrobial susceptibility testing showed: 100% sensitivity for amikacin; 83% for imipenem; 71% for cefotaxime; and 71% for trimethoprim-sulphamethoxazole. The disease remained localized in the lung in five cases, with a trend toward chronicity in one with bronchiectasis. In the other five, the disease disseminated, affecting subcutaneous tissue, the central nervous system and the kidney. Three patients died, one with disseminated disease and two who were receiving corticosteroid therapy. The following conclusions were reached: 1) pulmonary nocardiosis is difficult to diagnose, diagnosis is frequently delayed and a high level of suspicion is, thus, required in patients with underlying diseases or chronic corticosteroid therapy; 2) there is frequent dissemination and high mortality; and 3) antimicrobial combinations with proven synergy, such as imipenem and amikacin, are recommended for initial therapy.


E-Skin Bimodal Sensors for Robotics and Prosthesis Using PDMS Molds Engraved by Laser.
Electronic skin (e-skin) is pursued as a key component in robotics and prosthesis to confer them sensing properties that mimic human skin. For pressure monitoring, a great emphasis on piezoresistive sensors was registered due to the simplicity of sensor design and readout mechanism. For higher sensitivity, films composing these sensors may be micro-structured, usually by expensive photolithography techniques or low-cost and low-customizable molds. Sensors commonly present different sensitivities in different pressure ranges, which should be avoided in robotics and prosthesis applications. The combination of pressure sensing and temperature is also relevant for the field and has room for improvement. This work proposes an alternative approach for film micro-structuration based on the production of highly customizable and low-cost molds through laser engraving. These bimodal e-skin piezoresistive and temperature sensors could achieve a stable sensitivity of -6.4 × 10-3 kPa-1 from 1.6 kPa to 100 kPa, with a very robust and reproducible performance over 27,500 cycles of objects grasping and releasing and an exceptionally high temperature coefficient of resistance (TCR) of 8.3%/°C. These results point toward the versatility and high benefit/cost ratio of the laser engraving technique to produce sensors with a suitable performance for robotics and functional prosthesis.


A role for caveolin-1 in desmoglein binding and desmosome dynamics.
Desmoglein-2 (Dsg2) is a desmosomal cadherin that is aberrantly expressed in human skin carcinomas. In addition to its well-known role in mediating intercellular desmosomal adhesion, Dsg2 regulates mitogenic signaling that may promote cancer development and progression. However, the mechanisms by which Dsg2 activates these signaling pathways and the relative contribution of its signaling and adhesion functions in tumor progression are poorly understood. In this study we show that Dsg2 associates with caveolin-1 (Cav-1), the major protein of specialized membrane microdomains called caveolae, which functions in both membrane protein turnover and intracellular signaling. Sequence analysis revealed that Dsg2 contains a putative Cav-1-binding motif. A permeable competing peptide resembling the Cav-1 scaffolding domain bound to Dsg2, disrupted normal Dsg2 staining and interfered with the integrity of epithelial sheets in vitro. Additionally, we observed that Dsg2 is proteolytically processed; resulting in a 95-kDa ectodomain shed product and a 65-kDa membrane-spanning fragment, the latter of which localizes to lipid rafts along with full-length Dsg2. Disruption of lipid rafts shifted Dsg2 to the non-raft fractions, leading to the accumulation of these proteins. Interestingly, Dsg2 proteolytic products are elevated in vivo in skin tumors from transgenic mice overexpressing Dsg2. Collectively, these data are consistent with the possibility that accumulation of truncated Dsg2 protein interferes with desmosome assembly and/or maintenance to disrupt cell-cell adhesion. Furthermore, the association of Dsg2 with Cav-1 may provide a mechanism for regulating mitogenic signaling and modulating the cell-surface presentation of an important adhesion molecule, both of which could contribute to malignant transformation and tumor progression.


Nonlinear elimination kinetics of 5-fluoro-2'-deoxyuridine in isolated perfused rat liver and isolated hepatocytes.
The kinetic parameters of the cytostatic agent 5-fluoro-2'-deoxyluridine (FUDR) were studied in isolated rat hepatocytes and in the isolated perfused rat liver. In both experimental setups a dose dependency of the elimination parameters, half-life and clearance, was observed with a calculated turning point around 250 microM. In the medium of rat hepatocytes incubated at low (0.1 microM) to high (2000 microM) FUDR, the majority of the metabolites consisted of the catabolite alpha-fluoro-beta-alanine. The nucleobase metabolites, 5-fluorouracil and its primary product 5,6-dihydro-5-fluorouracil, approached apparent steady-state levels comprising 10 to 15% of the initial concentration. In the intracellular phase of hepatocytes incubated at 300 microM FUDR almost 90% of the FUDR-derived material was alpha-fluoro-beta-alanine, whereas essentially no unchanged FUDR could be detected. Similar results were obtained at extracellular FUDR concentrations exceeding 300 microM. In the isolated perfused rat liver, the clearance decreased to 15 to 20% of the corresponding values when the initial concentration was raised from 24 to 2400 microM. At the end of perfusion alpha-fluoro-beta-alanine comprised 90 to 95% of FUDR-derived total radioactivity in the tissue even at initially 2400 microM FUDR, although at this FUDR dosage 20% of the substrate remained unmetabolized in the medium. These results suggest that the limitation of hepatic FUDR elimination is not due to saturable hepatic metabolism but must be due to saturable uptake of these pyrimidine derivatives across the cellular membrane of parenchymal liver cells.


All-optical signal processing using dynamic Brillouin gratings.
The manipulation of dynamic Brillouin gratings in optical fibers is demonstrated to be an extremely flexible technique to achieve, with a single experimental setup, several all-optical signal processing functions. In particular, all-optical time differentiation, time integration and true time reversal are theoretically predicted, and then numerically and experimentally demonstrated. The technique can be exploited to process both photonic and ultra-wide band microwave signals, so enabling many applications in photonics and in radio science.


Activation of serum- and glucocorticoid-regulated protein kinase by agonists that activate phosphatidylinositide 3-kinase is mediated by 3-phosphoinositide-dependent protein kinase-1 (PDK1) and PDK2.
The PtdIns(3,4,5)P3-dependent activation of protein kinase B (PKB) by 3-phosphoinositide-dependent protein kinases-1 and -2 (PDK1 and PDK2 respectively) is a key event in mediating the effects of signals that activate PtdIns 3-kinase. The catalytic domain of serum- and glucocorticoid-regulated protein kinase (SGK) is 54% identical with that of PKB and, although lacking the PtdIns(3,4, 5)P3-binding pleckstrin-homology domain, SGK retains the residues that are phosphorylated by PDK1 and PDK2, which are Thr256 and Ser422 in SGK. Here we show that PDK1 activates SGK in vitro by phosphorylating Thr256. We also show that, in response to insulin-like growth factor-1 (IGF-1) or hydrogen peroxide, transfected SGK is activated in 293 cells via a PtdIns 3-kinase-dependent pathway that involves the phosphorylation of Thr256 and Ser422. The activation of SGK by PDK1 in vitro is unaffected by PtdIns(3,4,5)P3, abolished by the mutation of Ser422 to Ala, and greatly potentiated by mutation of Ser422 to Asp (although this mutation does not activate SGK itself). Consistent with these findings, the Ser422Asp mutant of SGK is activated by phosphorylation (probably at Thr256) in unstimulated 293 cells, and activation is unaffected by inhibitors of PtdIns 3-kinase. Our results are consistent with a model in which activation of SGK by IGF-1 or hydrogen peroxide is initiated by a PtdIns(3,4, 5)P3-dependent activation of PDK2, which phosphorylates Ser422. This is followed by the PtdIns(3,4,5)P3-independent phosphorylation at Thr256 that activates SGK, and is catalysed by PDK1. Like PKB, SGK preferentially phosphorylates serine and threonine residues that lie in Arg-Xaa-Arg-Xaa-Xaa-Ser/Thr motifs, and SGK and PKB inactivate glycogen synthase kinase-3 similarly in vitro and in co-transfection experiments. These findings raise the possibility that some physiological roles ascribed to PKB on the basis of the overexpression of constitutively active PKB mutants might be mediated by SGK.


Homophily and minority-group size explain perception biases in social networks.
People's perceptions about the size of minority groups in social networks can be biased, often showing systematic over- or underestimation. These social perception biases are often attributed to biased cognitive or motivational processes. Here we show that both over- and underestimation of the size of a minority group can emerge solely from structural properties of social networks. Using a generative network model, we show that these biases depend on the level of homophily, its asymmetric nature and on the size of the minority group. Our model predictions correspond well with empirical data from a cross-cultural survey and with numerical calculations from six real-world networks. We also identify circumstances under which individuals can reduce their biases by relying on perceptions of their neighbours. This work advances our understanding of the impact of network structure on social perception biases and offers a quantitative approach for addressing related issues in society.


Extensive DNA inversions in the B. fragilis genome control variable gene expression.
The obligately anaerobic bacterium Bacteroides fragilis, an opportunistic pathogen and inhabitant of the normal human colonic microbiota, exhibits considerable within-strain phase and antigenic variation of surface components. The complete genome sequence has revealed an unusual breadth (in number and in effect) of DNA inversion events that potentially control expression of many different components, including surface and secreted components, regulatory molecules, and restriction-modification proteins. Invertible promoters of two different types (12 group 1 and 11 group 2) were identified. One group has inversion crossover (fix) sites similar to the hix sites of Salmonella typhimurium. There are also four independent intergenic shufflons that potentially alter the expression and function of varied genes. The composition of the 10 different polysaccharide biosynthesis gene clusters identified (7 with associated invertible promoters) suggests a mechanism of synthesis similar to the O-antigen capsules of Escherichia coli.


[The Dutch PhD-factory: pivotal or research waste].
This commentary discusses the increase of the number of finished PhD-theses in the Netherlands among young medical doctors. I discuss their motivation, engagement, the lack of mentoring and the mental health symptoms that are highly prevalent among Dutch PhD students in university medical centers. As a consequence, the abovementioned factors may have a detrimental impact on the validity and integrity of research results. Furthermore, I discuss how the current academic hierarchy is shaping academic careers and dictating publication practices. PhD students are pivotal in fulfilling academic dreams of seniors by writing a considerable number of publications that help senior researchers increase their scientific output, advance their careers and help them to get tenured. I suggest that we should invest in sufficient, effective and responsible supervision and mentoring and should train our supervisors to become a responsible role model for their PhD students. For this we need different performance indicators, and more time and skills for seniors to invest in PhD students. As a result, supervisors will become more equipped and engaged in responsible supervision.


Characterization of antigen-specific CD4+ effector T cells in vivo: immunization results in a transient population of MEL-14-, CD45RB- helper cells that secretes interleukin 2 (IL-2), IL-3, IL-4, and interferon gamma.
In previous studies we demonstrated that, following activation by mitogens or alloantigens, helper T cell precursors proliferate and differentiate in vitro to produce a population of effector cells that secrete high titers of lymphokines upon restimulation. In this report, we demonstrate that a similar effector population develops in vivo following primary antigen stimulation. When restimulated with specific antigen in vitro, CD4+ T cells from mice primed 5 to 7 days previously by subcutaneous administration of keyhole limpet hemocyanin (KLH) in adjuvant, produced high levels of interleukin 2 (IL-2), IL-4, and IL-3, and little or no interferon gamma (IFN-gamma) or IL-5. The effector T cells provided excellent helper activity for in vitro antibody responses of 4-hydroxy-5-iodo-nitrophenyl acetic acid-primed B cells with the production principally of the immunoglobulin G1 (IgG1) and IgM isotypes, small quantities of IgG3, and no detectable IgG2a, or IgG2b. Antigen-specific secretion of IL-2, IL-3, and IL-4 by in vivo effectors was detectable by 12 hours following in vitro restimulation. IFN-gamma and IL-5 were not detected until 48 and 72 hours of culture, respectively, and low levels of these lymphokines were produced. Lymphokine production by primed CD4+ T cells could be induced as early as 3 days following immunization, peaked on day 5, and declined thereafter. The kinetics of in vivo appearance of effector CD4+ T cells that produce lymphokines upon restimulation in vitro were similar for each of the lymphokines examined. Mice depleted of precursor CD4+ T cells by adult thymectomy exhibited limited capacity to generate lymphokine secreting CD4+ T cells in response to primary immunization with KLH, suggesting that the majority of lymphokine producing T cells arise from short-lived and/or precursor cells. Separation of CD4+ T cells from KLH-primed mice on the basis of expression of the lymph node-specific homing receptor, MEL-14, revealed that antigen-specific production of IL-2, IL-3, IL-4, and IFN-gamma was exclusively associated with the MEL-14- subset of CD4+ T cells. Separation on the basis of CD45RB expression, demonstrated that antigen-specific lymphokine production was primarily associated with the minor CD45RB- population, which has been previously associated with memory activity. Our results indicate that primary in vivo immunization leads to the development of a transient population of helper-effectors with a unique phenotype that can produce large quantities of lymphokines and mediate excellent helper activity for B cells.(ABSTRACT TRUNCATED AT 400 WORDS)


UNC45A confers resistance to histone deacetylase inhibitors and retinoic acid.
To identify potential biomarkers of therapy response, we have previously done a large-scale gain-of-function genetic screen to identify genes whose expression confers resistance to histone deacetylase inhibitors (HDACI). This genetic screen identified two genes with a role in retinoic acid signaling, suggesting that HDACIs target retinoic acid signaling as part of their anticancer effect. We study here a third gene identified in this genetic screen, UNC45A, and assess its role in retinoic acid signaling and responses to HDACIs using cell-based proliferation and differentiation assays and transcriptional reporter gene assays. The vertebrate Unc45 genes are known for their roles in muscle development and the assembly and cochaperoning of the muscle motor protein myosin. Here, we report that human UNC45A (GCUNC45) can render transformed cells resistant to treatment with HDACIs. We show that UNC45A also inhibits signaling through the retinoic acid receptor alpha. Expression of UNC45A inhibits retinoic acid-induced proliferation arrest and differentiation of human neuroblastoma cells and inhibits the induction of endogenous retinoic acid receptor target genes. These data establish an unexpected role for UNC45A in causing resistance to both HDACI drugs and retinoic acid. Moreover, our data lend further support to the notion that HDACIs exert their anticancer effect, at least in part, through an effect on retinoic acid signaling.


Prevalence and risk factors of Plasmodium falciparum infections in pregnant women of Luanda, Angola.
Pregnant women are at increased risk of malaria, but in Angola, epidemiologic data from this group is almost inexistent. We conducted a cross-sectional study to determine the prevalence and risk factors of Plasmodium falciparum infections in 567 pregnant Angolan women living in Luanda province. One in five women had P. falciparum at delivery, diagnosed by PCR assay. Age, residence and history of malaria during pregnancy were significantly associated with P. falciparum infection, but gravidity and use of anti-malarial drugs were not. Placental infections were significantly more common in women ≤18 years old and in primigravidae, but we could not correlate placental infections with poor pregnancy outcomes. These findings are relevant to malaria control policies in Luanda, Angola.


Glutathione S-Transferase and Clusterin, New Players in the Ischemic Preconditioning Renal Protection in a Murine Model of Ischemia and Reperfusion.
Renal ischemia and reperfusion injury (IRI) involves oxidative stress, disruption of microvasculature due to endothelial cell damage, loss of epithelial cell polarity secondary to cytoskeletal alterations, inflammation, and the subsequent transition into a mesenchymal phenotype. Ischemic preconditioning (IPC) has been proposed as a therapeutic strategy to avoid/ameliorate the IRI. Since previous results showed that IPC could have differential effects in kidney cortex vs. kidney medulla, in the present study we analyzed the effectiveness and molecular mechanisms implicated in IPC in both kidney regions. We evaluated 3 experimental groups of BALB/c male mice: control (sham surgery); renal ischemia (30 min) by bilateral occlusion of the renal pedicle and reperfusion (48 hours) (I/R); and renal IPC (two cycles of 5 min of ischemia and 5 min of reperfusion) applied just before I/R. Acute kidney injury was evaluated by glomerular filtration rate (GFR), Neutrophil Gelatinase-Associated Lipocalin (NGAL) blood level, and histologic analysis. Oxidative stress was studied measurement the Glutathione S-Transferase (GST) activity, GSH/GSSG ratio, and lipoperoxidation levels. Inflammatory mediators (IL-1β, IL-6, Foxp3, and IL-10) were quantified by qRT-PCR. The endothelial (PECAM-1), epithelial (AQP-1), mesenchymal (Vimentin, Fascin, and Hsp47), iNOS, clusterin, and Hsp27 expression were evaluated (qRT-PCR and/or Western blot). The IPC protocol prevented the decrease of GFR, reduced the plasma NGAL, and ameliorated morphological damage in the kidney cortex after I/R. The IPC also prevented the downregulation of GST activity, lipoperoxidation and ameliorated the oxidized glutathione. In addition, IPC prevented the upregulation of vimentin, fascin, and Hsp47, which was associated with the prevention of the downregulation of AQP1 after I/R. The protective effect of IPC was associated with the upregulation of Hsp27, Foxp3, and IL-10 expression in the renal cortex. However, the upregulation of iNOS, IL-1β, IL-6, and clusterin by I/R were not modified by IPC. IPC conferred better protection in the kidney cortex as compared to the kidney medulla. The protective effect of IPC was associated with amelioration of oxidative stress, tubular damage, and the induction of markers of Treg lymphocytes activity in the cortical region. Further studies are needed to evaluate if lower tubular cell stress/damage after I/R may explain the preferential induction of Treg response in the kidney cortex induced by IPC.


Combining mutations in HIV-1 reverse transcriptase with mutations in the HIV-1 polypurine tract affects RNase H cleavages involved in PPT utilization.
The RNase H cleavages that generate and remove the polypurine tract (PPT) primer during retroviral reverse transcription must be specific to generate linear viral DNAs that are suitable substrates for the viral integrase. To determine if specific contacts between reverse transcriptase (RT) and the PPT are a critical factor in determining the cleavage specificity of RNase H, we made HIV-1 viruses containing mutations in RT and the PPT at the locations of critical contacts between the protein and the nucleic acid. The effects on titer and RNase H cleavage suggest that combining mutations in RT with mutations in the PPT affect the structure of the protein of the RT/nucleic acid complex in ways that affect the specificity and the rate of PPT cleavage. In contrast, the mutations in the PPT (alone) and RT (alone) affect the specificity of PPT cleavage but have much less effect on the overall rate of cleavage.


Discovering misattributed paternity in living kidney donation: prevalence, preference, and practice.
When evaluating a living kidney donor and recipient with a father-child relationship, it may be discovered that the two are not biologically related. We analyzed data from the United Network for Organ Sharing and the Canadian Organ Replacement Registry to determine how frequently this occurs. We surveyed 102 potential donors, recipients, and transplant professionals for their opinion on whether such information should be disclosed to the donor-recipient pair. We communicated with transplant professionals from 13 Canadian centers on current practices for handling this information. In the United States and Canada, the prevalence of father-child living kidney donor-recipient pairs with less than a one-haplotype human leukocyte antigen match (i.e., misattributed paternity) is between 1% and 3%, or approximately 0.25% to 0.5% of all living kidney donations. Opinions about revealing this information were variable: 23% strongly favored disclosure; whereas, 24% were strongly opposed to it. Current practices are variable; some centers disclose this information, whereas others do not. Discovering misattributed paternity in living donation is uncommon but can occur. Opinions on how to deal with this sensitive information are variable. Discussion among transplant professionals will facilitate best practices and policies. Strategies adopted by some centers can be considered.


Late Permian marine ecosystem collapse began in deeper waters: evidence from brachiopod diversity and body size changes.
Analysis of Permian-Triassic brachiopod diversity and body size changes from different water depths spanning the continental shelf to basinal facies in South China provides insights into the process of environmental deterioration. Comparison of the temporal changes of brachiopod diversity between deepwater and shallow-water facies demonstrates that deepwater brachiopods disappeared earlier than shallow-water brachiopods. This indicates that high environmental stress commenced first in deepwater settings and later extended to shallow waters. This environmental stress is attributed to major volcanic eruptions, which first led to formation of a stratified ocean and a chemocline in the outer shelf and deeper water environments, causing the disappearance of deep marine benthos including brachiopods. The chemocline then rapidly migrated upward and extended to shallow waters, causing widespread mass extinction of shallow marine benthos. We predict that the spatial and temporal patterns of earlier onset of disappearance/extinction and ecological crisis in deeper water ecosystems will be recorded during other episodes of rapid global warming.


Spinal muscular atrophy pathogenic mutations impair the axonogenic properties of axonal-survival of motor neuron.
The axonal survival of motor neuron (a-SMN) protein is a truncated isoform of SMN1, the spinal muscular atrophy (SMA) disease gene. a-SMN is selectively localized in axons and endowed with remarkable axonogenic properties. At present, the role of a-SMN in SMA is unknown. As a first step to verify a link between a-SMN and SMA, we investigated by means of over-expression experiments in neuroblastoma-spinal cord hybrid cell line (NSC34) whether SMA pathogenic mutations located in the N-terminal part of the protein affected a-SMN function. We demonstrated here that either SMN1 missense mutations or small intragenic re-arrangements located in the Tudor domain consistently altered the a-SMN capability of inducing axonal elongation in vitro. Mutated human a-SMN proteins determined in almost all NSC34 motor neurons the growth of short axons with prominent morphologic abnormalities. Our data indicate that the Tudor domain is critical in dictating a-SMN function possibly because it is an association domain for proteins involved in axon growth. They also indicate that Tudor domain mutations are functionally relevant not only for FL-SMN but also for a-SMN, raising the possibility that also a-SMN loss of function may contribute to the pathogenic steps leading to SMA.


Regulation of the hypoxia-inducible factor-1 alpha. ARNT is not necessary for hypoxic induction of HIF-1 alpha in the nucleus.
Hypoxia-inducible factor-1 (HIF-1) is a master regulator of mammalian oxygen homeostasis. HIF-1 consists of two subunits, HIF-1 alpha and the aryl hydrocarbon receptor nuclear translocator (ARNT). Whereas hypoxia prevents ubiquitination and proteasomal degradation of HIF-1 alpha, ARNT expression is thought to be oxygen-independent. We and others previously showed that ARNT is indispensable for HIF-1 DNA-binding and transactivation function. To examine the requirement of ARNT for accumulation and nuclear translocation of HIF-1 alpha in hypoxia, we used ARNT-mutant mouse hepatoma and ARNT-deficient embryonic stem cells. As shown by immunofluorescence, HIF-1 alpha accumulation in the nucleus of hypoxic cells did not require ARNT, demonstrating that nuclear translocation is intrinsic to HIF-1 alpha. During biochemical separation, both HIF-1 alpha and ARNT tend to leak from the nuclei in the absence of the corresponding subunit, suggesting that heterodimerization is required for stable association within the nuclear compartment. Nuclear stabilization of the heterodimer might also explain the hypoxically increased total cellular ARNT levels observed in some of the cell lines examined.


The role of a defunctioning stoma for colonic and perianal Crohn's disease in the biological era.
A defunctioning stoma is a therapeutic option for colonic or perianal Crohn's disease. In the pre-biologic era the response rate to defunctioning in our unit was high (86%), but intestinal continuity was only restored in 11-20%. Few data exist on the outcome of defunctioning since the widespread introduction of biologicals. All patients undergoing a defunctioning stoma for colonic/perianal Crohn's disease since 2003-2011 were identified from a prospective database. Indications for surgery, medical therapy, response to defunctioning and long-term clinical outcome were recorded. Successful restoration of continuity was defined as no stoma at last follow up. Seventy-six patients were defunctioned (57 with biologicals) and at last follow up, 20 (27%) had continuity restored. Early clinical response rate (<3 months) was 15/76 (20%) and overall response 31/76 (41%). Complex anal fistulae/stenosis were associated with a very low chance of restoring continuity (10% and 0%, respectively), while colitis was associated with a higher chance of restoring continuity (48%). Endoscopic or histological improvement in colitis after defunctioning was associated with a higher rate of restoring continuity (10/16, 63%) compared to no such improvement (4/15, 27%, p = 0.05). Those failing biologics had similar chance of restoration as those not receiving biologics, 15/57 (26%) and 5/19 (26%), respectively. Overall response to colonic defunctioning was 41%. Successful restoration of continuity occurred in 27%, but 48% in the absence of perianal disease. Response is appreciably less in the pre-biologic era, so patient and physician expectations need to be managed appropriately.


Twenty-eight days at 3454-m altitude diminishes respiratory capacity but enhances efficiency in human skeletal muscle mitochondria.
Modifications of skeletal muscle mitochondria following exposure to high altitude (HA) are generally studied by morphological examinations and biochemical analysis of expression. The aim of this study was to examine tangible measures of mitochondrial function following a prolonged exposure to HA. For this purpose, skeletal muscle biopsies were obtained from 8 lowland natives at sea level (SL) prior to exposure and again after 28 d of exposure to HA at 3454 m. High-resolution respirometry was performed on the muscle samples comparing respiratory capacity and efficiency. Exercise capacity was assessed at SL and HA. Respirometric analysis revealed that mitochondrial respiratory capacity diminished in complex I- and complex II-specific respiration in addition to a loss of maximal state-3 oxidative phosphorylation capacity from SL to HA, all independent from alterations in mitochondrial content. Leak control coupling, respiratory control ratio, and oligomycin-induced leak respiration, all measures of mitochondrial efficiency, improved in response to HA exposure. SL respiratory capacities correlated with measures of exercise capacity near SL, whereas mitochondrial efficiency correlated best with exercise capacity following HA. This data demonstrate that 1 mo of exposure to HA reduces respiratory capacity in human skeletal muscle; however, the efficiency of electron transport improves.


Institutional trust in the national social security and municipal healthcare systems for the elderly in Japan.
Japanese social security systems and institutions for the elderly, as well as the general attitude toward elderly care services among the Japanese population, have been undergoing a dramatic change. By examining the association between institutional trust, which is a representative element of social capital, and anxiety regarding receiving elderly care, we can uncover clues toward building a more robust social security system for the elderly. This study examines the relationship between institutional trust, in the national social security and municipal healthcare systems for the elderly, and anxiety with respect to receiving elderly care among the general Japanese population. A cross-sectional survey was conducted using a self-administered questionnaire that was returned by mail in January and February 2005. The target population was 4735 community residents aged 20-75 years, who lived in the city of Koka, Shiga, Japan. A total of 2264 questionnaires were included in the analysis. A binominal logistic regression analysis showed that responses of 'trust' [odds ratio (OR): 2.09, 95% confidence interval (95% CI): 1.01-4.30] and 'strongly trust' (OR: 3.80, 95% CI: 1.55-9.31) for the national system were associated with not having anxiety regarding elderly care, compared with the reference category of feeling strongly distrust. However, trust in the municipal system showed no association with this anxiety. These results indicate the importance of developing strategies to increase a common trust in the national care services for the elderly to reduce the anxiety people feel regarding whether they will be able to receive elderly care when required.


Life satisfaction of Taiwanese dental graduates received residencies in the U.S.: a cross-sectional study.
Each year, more than 200 international dental graduates start U.S. specialty trainings to become specialists. It is unknown if their life satisfaction is associated with any dental career-related factor before residencies (e.g. dental school class rank, research experience, or private practice experience) and after residencies (e.g. staying in the U.S., teaching status, workplace, or board certification). This cross-sectional study aimed to identify these potential factors by surveying Taiwanese dental graduates who pursued U.S. residencies. Life satisfaction was measured with a structured questionnaire, Satisfaction With Life Scale (SWLS), which includes five statements on a 5-point Likert scale. Online surveys were sent out to 290 Taiwanese dental graduates who were known to pursue U.S. residencies. T-test, one way analysis of variance, and multivariable adjusted generalized linear model (GLM) were used to assess the differences of mean SWLS scores from different variables. Surveys were completed by 158 dentists. Mean SWLS score of 125 specialists was higher (p = 0.0007) than the score of 33 residents. For the 125 specialists, multivariable adjusted GLM demonstrated better life satisfaction was positively associated with multiple independent factors, such as having research experience, being ranked in the top 26 ~ 50% of the class in dental school, starting U.S. residency within 4 years after dental school, starting residency before year 1996, and specializing in endodontics (vs. periodontics). Life satisfaction was not associated with any factors after residency (e.g. staying in the U.S. afterwards, teaching status, or workplace), but better mean life satisfaction score was significantly associated with being American specialty board certified (p < 0.001) for the specialists in the 26 ~ 75% of their class in dental school. For the 33 residents, better mean life satisfaction score was associated with better dental school class rank in both bivariate (p = 0.020) and multivariable adjusted GLM (p = 0.004) analyses. The life satisfaction of Taiwanese dental graduates pursuing U.S. residencies might be associated with some professional factors, such as research experience, dental school class rank, residency timing, specialty type, and specialty board certification. We hope our results may provide some objective information on making career decisions for international dental graduates/students who are preparing for U.S. residency.


Manganese and aging.
Manganese (Mn) is an essential metal that is required as a cofactor for many enzymes and is necessary for optimal biological function. Mn is abundant in the earth's crust and is present in soil and well water. Mn is also found in industrial settings, including mining, welding, and battery manufacture. Mn is also present in infant formula, parenteral nutrition, as well as pesticides and gasoline additives. A sufficient amount of Mn is obtained from most diets, and Mn deficiency is exceedingly rare. Excessive exposure to Mn in high doses can result in a condition known as manganism that results in psychological and emotional disturbances and motor symptoms that are reminiscent of Parkinson's disease, including gait disturbance, tremor, rigidity, and bradykinesia. Treatment for manganism is to remove the patient from Mn exposure, though symptoms are generally irreversible. The effects of exposure to Mn at lower doses are less clear. Little work has been done to evaluate the effects of chronic exposure to subclinical levels of Mn, especially in regard to lifelong exposures and the effects on the aging process. Mn is known to have effects on some of the same mechanistic processes that are altered in aging. This review will describe the general effects of Mn exposure and will focus on how Mn may be related to some of the mechanism of aging: neurogenesis, oxidative stress, and microglial activation and inflammation.


The comparative effectiveness of decision aids in diverse populations with early stage prostate cancer: a study protocol for a cluster-randomized controlled trial in the NCI Community Oncology Research Program (NCORP), Alliance A191402CD.
Treatments for localized prostate cancer present challenging tradeoffs in the face of uncertain treatment benefits. These options are best weighed in a process of shared decision-making with the patient's healthcare team. Minority men experience disparities in prostate cancer outcomes, possibly due in part to a lack of optimal communication during treatment selection. Decision aids facilitate shared decision-making, improve knowledge of treatment options, may increase satisfaction with treatment choice, and likely facilitate long-term quality of life. This study will compare the effect of two evidence-based decision aids on patient knowledge and on quality of life measured one year after treatment, oversampling minority men. One decision aid will be administered prior to specialist consultation, preparing patients for a treatment discussion. The other decision aid will be administered within the consultation to facilitate transparent, preference-sensitive, and evidence-informed deliberations. The study will utilize a four-arm, block-randomized design to test whether each decision aid alone (Arms 1 and 2) or in combination (Arm 3) can improve patient knowledge and quality of life compared to usual care (Arm 4). The study, funded by the National Cancer Institute's Community Oncology Research Program (NCORP), will be deployed within select institutions that have demonstrated capacity to recruit minority populations into urologic oncology trials. Upon completion of the trial, we will have 1) tested the effectiveness of two evidence-based decision aids in enhancing patients' knowledge of options for prostate cancer therapy and 2) estimated whether decision aids may improve patient quality of life one year after initial treatment choice. Clinicaltrials.gov: NCT03103321 . The trial registration date (on ClinicalTrials.gov) was April 6, 2017.


Juxtanodin is an intrinsically disordered F-actin-binding protein.
Juxtanodin, also called ermin, is an F-actin-binding protein expressed by oligodendrocytes, the myelin-forming cells of the central nervous system. While juxtanodin carries a short conserved F-actin-binding segment at its C terminus, it otherwise shares no similarity with known protein sequences. We carried out a structural characterization of recombinant juxtanodin in solution. Juxtanodin turned out to be intrinsically disordered, as evidenced by conventional and synchrotron radiation CD spectroscopy. Small-angle X-ray scattering indicated that juxtanodin is a monomeric, highly elongated, unfolded molecule. Ensemble optimization analysis of the data suggested also the presence of more compact forms of juxtanodin. The C terminus was a strict requirement for co-sedimentation of juxtanodin with microfilaments, but juxtanodin had only mild effects on actin polymerization. The disordered nature of juxtanodin may predict functions as a protein interaction hub, although F-actin is its only currently known binding partner.


A unique secreted adenovirus E3 protein binds to the leukocyte common antigen CD45 and modulates leukocyte functions.
The E3 transcription unit of human adenoviruses (Ads) encodes immunomodulatory proteins. Interestingly, the size and composition of the E3 region differs considerably among Ad species, suggesting that distinct sets of immunomodulatory E3 proteins may influence their interaction with the human host and the disease pattern. However, to date, only common immune evasion functions of species C E3 proteins have been described. Here we report on the immunomodulatory activity of a species D-specific E3 protein, E3/49K. Unlike all other E3 proteins that act on infected cells, E3/49K seems to target uninfected cells. Initially synthesized as an 80- to 100-kDa type I transmembrane protein, E3/49K is subsequently cleaved, with the large ectodomain (sec49K) secreted. We found that purified sec49K exhibits specific binding to lymphoid cell lines and all primary leukocytes, but not to fibroblasts or epithelial cells. Consistent with this binding profile and the molecular mass, the sec49K receptor was identified as the cell surface protein tyrosine phosphatase CD45. Antibody-blocking studies suggested that sec49K binds to the membrane proximal domains present in all CD45 isoforms. Functional studies showed that sec49K can suppress the activation and cytotoxicity of natural killer cells as well as the activation, signaling, and cytokine production of T cells. Thus, we have discovered an adenovirus protein that is actively secreted and describe immunomodulatory activities of an E3 protein uniquely expressed by a single Ad species.


Does the National Institutes of Health Stroke Scale favor left hemisphere strokes? NINDS t-PA Stroke Study Group.
The National Institutes of Health Stroke Scale (NIHSS) is a valid, reproducible scale that measures neurological deficit. Of 42 possible points, 7 points are directly related to measurement of language compared with only 2 points related to neglect. We examined the placebo arm of the NINDS t-PA stroke trial to test the hypothesis that the total volume of cerebral infarction in patients with right hemisphere strokes would be greater than the volume of cerebral infarction in patients with left hemisphere strokes who have similar NIHSS scores. The volume of stroke was determined by computerized image analysis of CT films and CT images stored on computer tape and optical disks. Cube-root transformation of lesion volume was performed for each CT. Transformed lesion volume was analyzed in a logistic regression model to predict volume of stroke by NIHSS score for each hemisphere. Spearman rank correlation was used to determine the relation between the NIHSS score and lesion volume. The volume for right hemisphere stroke was statistically greater than the volume for left hemisphere strokes, adjusting for the baseline NIHSS (P<0. 001). For each 5-point category of the NIHSS score <20, the median volume of right hemisphere strokes was approximately double the median volume of left hemisphere strokes. For example, for patients with a left hemisphere stroke and a 24-hour NIHSS score of 16 to 20, the median volume of cerebral infarction was 48 mL (interquartile range 14 to 111 mL) as compared with 133 mL (interquartile range 81 to 208 mL) for patients with a right hemisphere stroke (P<0.001). The median volume of a right hemisphere stroke was roughly equal to the median volume of a left hemisphere stroke in the next highest 5-point category of the NIHSS. The Spearman rank correlation between the 24-hour NIHSS score and 3-month lesion volume was 0.72 for patients with left hemisphere stroke and 0.71 for patients with right hemisphere stroke. For a given NIHSS score, the median volume of right hemisphere strokes is consistently larger than the median volume of left hemisphere strokes. The clinical implications of our finding need further exploration.


Potential for entomopathogenic nematodes in biological control: a meta-analytical synthesis and insights from trophic cascade theory.
Entomopathogenic nematodes (EPN) are ubiquitous and generalized consumers of insects in soil food webs, occurring widely in natural and agricultural ecosystems on six continents. Augmentative releases of EPN have been used to enhance biological control of pests in agroecosystems. Pest managers strive to achieve a trophic cascade whereby natural-enemy effects permeate down through the food web to suppress host herbivores and increase crop production. Although trophic cascades have been studied in diverse aboveground arthropod-based systems, they are infrequently investigated in soil systems. Moreover, no overall quantitative assessment of the effectiveness of EPN in suppressing hosts with cascading benefits to plants has been made. Toward synthesizing the available but limited information on EPN and their ability to suppress prey and affect plant yield, we surveyed the literature and performed a meta-analysis of 35 published studies. Our analysis found that effect sizes for arthropod hosts as a result of EPN addition were consistently negative and indirect effects on plants were consistently positive. Results held across several different host metrics (abundance, fecundity and survival) and across measures of plant performance (biomass, growth, yield and survival). Moreover, the relationship between plant and host effect sizes was strikingly and significantly negative. That is, the positive impact on plant responses generally increased as the negative effect of EPN on hosts intensified, providing strong support for the mechanism of trophic cascades. We also review the ways in which EPN might interact antagonistically with each other and other predators and pathogens to adversely affect host suppression and dampen trophic cascades. We conclude that the food web implications of multiple-enemy interactions involving EPN are little studied, but, as management techniques that promote the long-term persistence of EPN are improved, antagonistic interactions are more likely to arise. We hope that the likely occurrence of antagonistic interactions in soil food webs should stimulate researchers to conduct field experiments explicitly designed to examine multiple-enemy interactions involving EPN and their cascading effects to hosts and plants.


Chest physiotherapy in cystic fibrosis: a comparative study of autogenic drainage and the active cycle of breathing techniques with postural drainage.
BACKGROUND--Autogenic drainage has been suggested as an alternative method of chest physiotherapy in patients with cystic fibrosis. In this study autogenic drainage was compared with the active cycle of breathing techniques (ACBT) together with postural drainage. METHODS--Eighteen patients with cystic fibrosis took part in a randomised two-day crossover trial. There were two sessions of one method of physiotherapy on each day, either autogenic drainage or ACBT. The study days were one week apart. On each day the patients were monitored for six hours. Mucus movement was quantified by a radioaerosol technique. Airway clearance was studied qualitatively using xenon-133 scintigraphic studies at the start and end of each day. Expectorated sputum was collected during and for one hour after each session of physiotherapy. Pulmonary functions tests were performed before and after each session. Oxygen saturation (SaO2) and heart rate were measured before, during, and after each session. RESULTS--Autogenic drainage cleared mucus from the lungs faster than ACBT over the whole day. Both methods improved ventilation, as assessed by the xenon-133 ventilation studies. No overall differences were found in the pulmonary function test results, but more patients had an improved forced expiratory flow from 25% to 75% with autogenic drainage, while more showed an improved forced vital capacity with ACBT. No differences were found in sputum weight and heart rate, nor in mean SaO2 over the series, but four patients desaturated during ACBT. CONCLUSIONS--Autogenic drainage was found to be as good as ACBT at clearing mucus in patients with cystic fibrosis and is therefore an effective method of home physiotherapy. Patients with cystic fibrosis should be assessed as to which method suits them best.


Elevated baseline serum glutamate as a pharmacometabolomic biomarker for acamprosate treatment outcome in alcohol-dependent subjects.
Acamprosate has been widely used since the Food and Drug Administration approved the medication for treatment of alcohol use disorders (AUDs) in 2004. Although the detailed molecular mechanism of acamprosate remains unclear, it has been largely known that acamprosate inhibits glutamate action in the brain. However, AUD is a complex and heterogeneous disorder. Thus, biomarkers are required to prescribe this medication to patients who will have the highest likelihood of responding positively. To identify pharmacometabolomic biomarkers of acamprosate response, we utilized serum samples from 120 alcohol-dependent subjects, including 71 responders (maintained continuous abstinence) and 49 non-responders (any alcohol use) during 12 weeks of acamprosate treatment. Notably, baseline serum glutamate levels were significantly higher in responders compared with non-responders. Importantly, serum glutamate levels of responders are normalized after acamprosate treatment, whereas there was no significant glutamate change in non-responders. Subsequent functional studies in animal models revealed that, in the absence of alcohol, acamprosate activates glutamine synthetase, which synthesizes glutamine from glutamate and ammonia. These results suggest that acamprosate reduces serum glutamate levels for those who have elevated baseline serum glutamate levels among responders. Taken together, our findings demonstrate that elevated baseline serum glutamate levels are a potential biomarker associated with positive acamprosate response, which is an important step towards development of a personalized approach to treatment for AUD.


Integrative approach reveals composition of endoparasitoid wasp venoms.
The fruit fly Drosophila melanogaster and its endoparasitoid wasps are a developing model system for interactions between host immune responses and parasite virulence mechanisms. In this system, wasps use diverse venom cocktails to suppress the conserved fly cellular encapsulation response. Although numerous genetic tools allow detailed characterization of fly immune genes, lack of wasp genomic information has hindered characterization of the parasite side of the interaction. Here, we use high-throughput nucleic acid and amino acid sequencing methods to describe the venoms of two related Drosophila endoparasitoids with distinct infection strategies, Leptopilina boulardi and L. heterotoma. Using RNA-seq, we assembled and quantified libraries of transcript sequences from female wasp abdomens. Next, we used mass spectrometry to sequence peptides derived from dissected venom gland lumens. We then mapped the peptide spectral data against the abdomen transcriptomes to identify a set of putative venom genes for each wasp species. Our approach captured the three venom genes previously characterized in L. boulardi by traditional cDNA cloning methods as well as numerous new venom genes that were subsequently validated by a combination of RT-PCR, blast comparisons, and secretion signal sequence search. Overall, 129 proteins were found to comprise L. boulardi venom and 176 proteins were found to comprise L. heterotoma venom. We found significant overlap in L. boulardi and L. heterotoma venom composition but also distinct differences that may underlie their unique infection strategies. Our joint transcriptomic-proteomic approach for endoparasitoid wasp venoms is generally applicable to identification of functional protein subsets from any non-genome sequenced organism.


Dimerization of Fab fragments enables ready screening of phage antibodies that affect hepatocyte growth factor/scatter factor activity on target cells.
A number of applications of antibodies in diagnosis and therapy require multivalent reagents either because of the polymeric nature of the antigens or because biological activity depends on an effect on the formation of homodimeric species. Here, we report a procedure for mass screening of phage-derived monomeric antibody fragments that depend on valency for activity. As a model system, a set of 13 phage-derived human Fab fragments were first selected against mouse and human recombinant hepatocyte growth factor/scatter factor (HGF/SF), a high molecular weight polypeptide growth factor related to the blood protease plasminogen and involved in development and cancer. These Fab fragments were subsequently screened for an effect on HGF/SF activity either as monomeric fragments or after dimerization with a monoclonal antibody (9E10) directed against a peptide tag on the fragments. Fab were identified that either inhibited or enhanced HGF/SF activity on target cell lines, but dimerization was required for this effect. The approach proposed should facilitate mass screening of phage-derived antibody fragments that depend on multiple valency for activity.


Subclinical atherosclerosis in northern and southern China: the Chinese paradox.
The incidence of coronary heart disease (CHD) is higher in Northern than that in Southern China, however differences in traditional CHD risk factors do not fully explain this. No study has examined the differences in subclinical atherosclerosis that may help explain the differences in incidence. This study examined these differences in subclinical atherosclerosis using coronary computed tomography (CT) for calcification between the Northern and Southern China. We selected a random sample of participants in a large multi-center ongoing epidemiologic study for coronary calcium scanning in one northern city (North) (Beijing, n = 49) and in two southern cities (South) (Shanghai, n = 50, and Guangzhou, n = 50). Participants from the three field centers (mean age 67 years) underwent coronary risk factor evaluation and cardiac CT scanning for coronary calcium measurement using the Multi-Ethnic Study of Atherosclerosis scanning protocol. Adjusted log-transformed coronary artery calcium score in North China (Beijing) was 3.1 ± 0.4 and in South China (Shanghai and Guangzhou) was 2.2 ± 0.3 (P = 0.04). Mean calcium score for the northern city of Beijing was three times higher than that of the southern city of Guangzhou (P = 0.01) and 2.5 times higher than for the southern city of Shanghai (P = 0.03). The extent of subclinical atherosclerosis is significantly higher in the northern city of Beijing than that in the two southern cities of Guangzhou and Shanghai, even after adjusting for standard cardiac risk factors. This finding suggests that standard risk factors do not fully explain north south differences in clinical CHD incidence.


Spontaneous regression of advanced cancer: identification of a unique genetically determined, age-dependent trait in mice.
We have established and studied a colony of mice with a unique trait of host resistance to both ascites and solid cancers induced by transplantable cells. One dramatic manifestation of this trait is age-dependent spontaneous regression of advanced cancers. This powerful resistance segregates as a single-locus dominant trait, is independent of tumor burden, and is effective against cell lines from multiple types of cancer. During spontaneous regression or immediately after exposure, cancer cells provoke a massive infiltration of host leukocytes, which form aggregates and rosettes with tumor cells. The cytolytic destruction of cancer cells by innate leukocytes is rapid and specific without apparent damage to normal cells. The mice are healthy and cancer-free and have a normal life span. These observations suggest a previously unrecognized mechanism of immune surveillance, which may have potential for therapy or prevention of cancer.