Reducing Food Insecurity and Improving Fruit and Vegetable Intake Among Farmers' Market Incentive Program Participants.
To determine whether participation in a farmers' market incentive pilot program had an impact on food security and fruit and vegetable (F&V) intake of participants. Participants in the Supplemental Nutrition Assistance Program were eligible to receive a dollar-per-dollar match up to $10/wk in farmers' market incentives. The researchers used a pretest-posttest design to measure F&V intake and food security status of 54 adult participants before and after receiving farmers' market incentives. The 6-item Behavior Risk Factor Surveillance System questionnaire and US Household Food Security Survey Module were used to measure F&V intake and food security, respectively. Wilcoxon signed-rank test was used to compare scores of F&V intake. After receiving incentives, fewer individuals reported experiencing food insecurity-related behaviors. A significantly increased intake (P < .05) was found among selected vegetables. Participation in a farmers' market incentive program was positively related to greater food security and intake of select vegetables among participants in the Supplemental Nutrition Assistance Program.


Allgrove syndrome with prominent neurological symptoms. Case Report.
We report a young woman with the clinical picture of Allgrove syndrome in whom neurological symptoms are prominent. It usually presents in the first decade of life with a deficiency of tears, recurrent vomiting and dysphagia due to achalasia, severe hypoglycemic seizures and shock due to adrenal insufficiency. Neurological symptoms such as hyperreflexia, dysarthria, hypernasal speech, ataxia, sensory impairment, muscle weakness, and mental retardation are extremely slow to develop and manifest at a later age. Diagnosis was based on clinical presentation and laboratory findings. She is the first patient from the Czech Republic with genetic confirmation of Allgrove syndrome. This patient is one of about 100 cases described in the literature and one of the few patients with all the main typical clinical features.


Time-resolved fluorescence imaging for specific and quantitative immunodetection of human kallikrein 2 and prostate-specific antigen in prostatic tissue sections.
To design protocols for specific and quantitative immunohistochemical detection of human kallikrein 2 (hK2) using lanthanide chelate-labeled monoclonal antibodies (Mabs) and time-resolved fluorescence imaging. Anti-prostate-specific antigen (PSA) Mabs were tested in microtiterplate assays for their ability to prevent PSA from cross-reacting with the anti-hK2 Mab 6H10. Europium-labeled 6H10 and terbium-labeled anti-PSA Mab 2E9, selected as the best blocker antibody, were used for dual-label immunodetection in routinely fixed benign (n = 7) and malignant (n = 5) prostate specimens. The amounts of IgG bound in tissue were calculated from drops containing known Mab concentrations. The use of anti-PSA Mab 2E9 for blocking diminished the cross-reaction from 5% to 0.3%. In the analyzed tissues, there was considerable variation in staining intensity for both proteins; PSA signals varied from 0.1 to 36.6 times that of hK2, with on average 10-fold more bound anti-PSA Mab than anti-hK2 Mab. In malignant tissue, the amounts of bound IgGs were lower and more variable than in benign tissue using both the anti-PSA Mab and the anti-hK2 Mab. The variation in signal intensities for PSA and hK2 correlated significantly in benign tissue (P >0.05), but not in benign hyperplastic and malignant specimens (P <0.05). Quantification of two lanthanide chelate-labeled antibodies bound in the same tissue section enabled comparison of PSA and hK2 content in individual cells. The average cellular content of hK2 relative to that of PSA was consistent with previous mRNA studies. The time-resolved fluorescence imaging-based quantification method has universal applicability in fixed tissue specimens.


Potential biosignatures in super-Earth atmospheres II. Photochemical responses.
Spectral characterization of super-Earth atmospheres for planets orbiting in the habitable zone of M dwarf stars is a key focus in exoplanet science. A central challenge is to understand and predict the expected spectral signals of atmospheric biosignatures (species associated with life). Our work applies a global-mean radiative-convective-photochemical column model assuming a planet with an Earth-like biomass and planetary development. We investigated planets with gravities of 1g and 3g and a surface pressure of 1 bar around central stars with spectral classes from M0 to M7. The spectral signals of the calculated planetary scenarios have been presented by in an earlier work by Rauer and colleagues. The main motivation of the present work is to perform a deeper analysis of the chemical processes in the planetary atmospheres. We apply a diagnostic tool, the Pathway Analysis Program, to shed light on the photochemical pathways that form and destroy biosignature species. Ozone is a potential biosignature for complex life. An important result of our analysis is a shift in the ozone photochemistry from mainly Chapman production (which dominates in Earth's stratosphere) to smog-dominated ozone production for planets in the habitable zone of cooler (M5-M7)-class dwarf stars. This result is associated with a lower energy flux in the UVB wavelength range from the central star, hence slower planetary atmospheric photolysis of molecular oxygen, which slows the Chapman ozone production. This is important for future atmospheric characterization missions because it provides an indication of different chemical environments that can lead to very different responses of ozone, for example, cosmic rays. Nitrous oxide, a biosignature for simple bacterial life, is favored for low stratospheric UV conditions, that is, on planets orbiting cooler stars. Transport of this species from its surface source to the stratosphere where it is destroyed can also be a key process. Comparing 1g with 3g scenarios, our analysis suggests it is important to include the effects of interactive chemistry.


Neuronavigated transcranial magnetic stimulation suggests that area V2 is necessary for visual awareness.
The primary visual cortex (V1) has been shown to be critical for visual awareness, but the importance of other low-level visual areas has remained unclear. To clarify the role of human cortical area V2 in visual awareness, we applied transcranial magnetic stimulation (TMS) over V2 while participants were carrying out a visual discrimination task and rating their subjective awareness. Individual retinotopic maps and modelling of the TMS-induced electric field in V1, V2 and V3d ensured that the electric field was at or under the phosphene threshold level in V1 and V3d, whereas in V2 it was at the higher suppressive level. As earlier shown for the V1, our results imply that also V2 is necessary for conscious visual experience. Visual awareness of stimulus presence was completely suppressed when the TMS pulse was delivered 44-84 ms after the onset of visual stimulus. Visual discrimination and awareness of stimulus features was impaired when the TMS pulse was delivered 44-104 ms after the visual stimulus onset. These results suggest that visual awareness cannot be generated without an intact V2.


Glycemic control in cardiac surgery: implementing an evidence-based insulin infusion protocol.
Acute hyperglycemia following cardiac surgery increases the risk of deep sternal wound infection, significant early morbidity, and mortality. Insulin infusion protocols that target tight glycemic control to treat hyperglycemia have been linked to hypoglycemia and increased mortality. Recently published studies examining glycemic control in critical illness and clinical practice guidelines from professional organizations support moderate glycemic control. To measure critical care nurses' knowledge of glycemic control in cardiac surgery before and after education. To evaluate the safety and effectiveness of an evidence-based insulin infusion protocol targeting moderate glycemic control in cardiac surgery patients. This evidence-based practice change was implemented in the cardiovascular unit in a community teaching hospital. Nurses completed a self-developed questionnaire to measure knowledge of glycemic control. Blood glucose data, collected (retrospectively) from anesthesia end time through 11:59 PM on postoperative day 2, were compared from 2 months before to 2 months after the practice change. Nurses' knowledge (test scores) increased significantly after education (pretest mean = 53.10, SD = 11.75; posttest mean = 79.10, SD = 12.02; t54 = -8.18, P < .001). Mean blood glucose level after implementation was 148 mg/dL. The incidence of hypoglycemia, 2.09% before and 0.22% after the intervention, was significantly reduced ( $${\hbox{ \chi }}_{1}^{2}$$ [n = 29] = 13.9, P < .001). The percentage of blood glucose levels less than 180 mg/dL was 88.30%. Increasing nurses' knowledge of glycemic control and implementing an insulin infusion protocol targeting moderate glycemic control were effective for treating acute hyperglycemia following cardiac surgery with decreased incidence of hypoglycemia.


Pitfalls in diagnosing coeliac disease.
To highlight the pitfalls in the diagnosis of coeliac disease and to make recommendations for its diagnosis and the management of refractory cases with equivocal histology. Six patients, referred since 1989 with a diagnosis of coeliac disease based on duodenal biopsy specimens taken at endoscopy, and who failed to respond to a gluten-free diet were studied. All patients were subjected to peroral jejunal biopsy. Morphometric analysis of villus height:crypt depth ratios, surface enterocyte cell heights, and intraepithelial counts was used to aid in the assessment of equivocal histology. Subsequent small intestinal biopsy specimens both taken when the patients were following a gluten-free diet and after gluten challenge were normal in all cases. Morphometric analysis and intraepithelial counts were normal. Misinterpretation of the original slides was often due to poor sample quality and tangential sectioning. Failure to respond to a gluten-free diet should always raise doubt regarding the initial diagnosis, especially when the findings are normal. For correct diagnosis at least three distal duodenal biopsy specimens should be taken simultaneously, and these should be of an adequate size and correctly orientated. Review by a histopathologist experienced in gastrointestinal diagnosis is essential in difficult cases. Quantitative morphometric analysis is helpful in equivocal cases, and jejunal suction biopsy, following a gluten challenge, may be necessary in patients refractory to treatment.


Genetic modifiers of chromatin acetylation antagonize the reprogramming of epi-polymorphisms.
Natural populations are known to differ not only in DNA but also in their chromatin-associated epigenetic marks. When such inter-individual epigenomic differences (or "epi-polymorphisms") are observed, their stability is usually not known: they may or may not be reprogrammed over time or upon environmental changes. In addition, their origin may be purely epigenetic, or they may result from regulatory variation encoded in the DNA. Studying epi-polymorphisms requires, therefore, an assessment of their nature and stability. Here we estimate the stability of yeast epi-polymorphisms of chromatin acetylation, and we provide a genome-by-epigenome map of their genetic control. A transient epi-drug treatment was able to reprogram acetylation variation at more than one thousand nucleosomes, whereas a similar amount of variation persisted, distinguishing "labile" from "persistent" epi-polymorphisms. Hundreds of genetic loci underlied acetylation variation at 2,418 nucleosomes either locally (in cis) or distantly (in trans), and this genetic control overlapped only partially with the genetic control of gene expression. Trans-acting regulators were not necessarily associated with genes coding for chromatin modifying enzymes. Strikingly, "labile" and "persistent" epi-polymorphisms were associated with poor and strong genetic control, respectively, showing that genetic modifiers contribute to persistence. These results estimate the amount of natural epigenomic variation that can be lost after transient environmental exposures, and they reveal the complex genetic architecture of the DNA-encoded determinism of chromatin epi-polymorphisms. Our observations provide a basis for the development of population epigenetics.


Mitral valve prolapse. Two-dimensional echocardiographic and angiographic correlation.
In order to define baseline descriptive criteria for the diagnosis of mitral valve prolapse with cross-sectional echocardiography, 49 patients undergoing catheterization were examined by a real-time, two-dimensional phased array echocardiographic imaging system. Angiography was used to separate patients into two distinct groups: 15 with normal mitral valve function and 34 with definite mitral valve prolapse. Systolic mitral leaflet and annulus motion were then observed in each patient and similarities and differences were noted between the two groups of patients. Correlative M-mode echocardiographic data were available in 37 patients. Certain two-dimensional echocardiographic findings restricted to the angiographically proven mitral valve prolapse group were defined: 1) posteriorly displaced coaptation of the leaflets, 2) systolic superior movement of one or both mitral leaflets above the level of the mitral ring, and 3) a systolic curling motion of the posterior mitral ring on its adjacent myocardium. One or more of these abnormalities were found in all 34 patients with angiographic mitral valve prolapse. When mitral valve prolapse does occur, the results of two-dimensional echocardiography would suggest that both leaflets are usually involved.


Cancer risk is not increased in patients treated for orthopaedic diseases with autologous bone marrow cell concentrate.
There is concern that regenerative cell-based therapies could result in increased risk of tumor formation. We investigated the long-term risks for systemic and site-specific cancers in patients who had received autologous bone marrow-derived stromal progenitor cells to treat orthopaedic lesions. A total of 1873 patients were treated from 1990 to 2006 with bone marrow-derived concentrated cells. Patients were monitored for cancer incidence from the date of the first operation (1990) until death, or until December 31, 2011. The mean follow-up time was 12.5 years (range, five to twenty-two years). The average number of colony-forming unit fibroblasts returned to the patients was 483,000 fibroblasts (range, 62,000 to 2,095,000 fibroblasts). The primary outcome was to evaluate with radiographs and/or magnetic resonance imaging the risk of tumorigenesis at the cell therapy treatment sites. The secondary outcome was to evaluate the risk of cancer diagnosed in areas other than the treatment site during the follow-up period. The relative risk of cancer was expressed as the ratio of observed and expected number of cases, that is, the standardized incidence ratio, according to the cancer incidence in the French population. No tumor formation was found at the treatment sites on the 7306 magnetic resonance images and 52,430 radiographs among the 1873 patients. Fifty-three cancers were diagnosed in areas other than the treatment site. On the basis of cancer incidence in the general population during the same period, the expected number of cancers was between ninety-seven and 108 for the same age and sex distribution. The range of the standardized incidence ratio for the follow-up period was between 0.49 and 0.54 (95% confidence interval, 0.30 to 0.80). This study found no increased cancer risk in patients after application of autologous cell-based therapy using bone marrow-derived stromal progenitor cells either at the treatment site or elsewhere in the patients after an average follow-up period of 12.5 years.


Various varying variances: The challenge of nuisance parameters to the practising biostatistician.
The 1997 Biometrics paper by Mike Kenward and James Roger has become a citation classic (more than 1260 citations by End June 2013 according to Google Scholar) and the solution that they proposed to deal with the problem of significance tests of fixed effects in REML models is now incorporated in many software packages and accepted by all biostatisticians as the method of choice. Nevertheless, it does not solve all problems, since there is more to analysis than just significance and since the problems that models with more than one variance pose arise in many contexts. In this paper, I discuss some problems and applications and make some tentative suggestions as to how they may be tackled. My excuse for raising problems I do not solve is that it may inspire James and Mike to complete what they started.


Electrochemical analysis of a novel ferrocene derivative as a potential antitumor drug.
Ferrocenes represent an interesting group of drugs with potential antitumor properties. Moreover, their electronic properties make them suitable for electrochemical studies. We determined an uptake of a novel ferrocene derivative in low μM concentrations by selected cancer cell lines and showed its localization predominantly in cytoplasm, using glassy carbon electrodes.


Correlation between surface electromyography and kinematics of the hindlimb of horses at trot on a treadmill.
The purpose of this study was to demonstrate the feasibility of surface electromyography in the horse and to correlate electromyographic activity with kinematic data. Surface electromyography of seven hindlimb muscles was recorded in five horses at trot on a treadmill. Simultaneously, kinematic analysis of the hindlimb was performed using a three-dimensional system and a unidirectional accelerometer was attached to the hoof. Electromyographic activities of the gluteus medius, vastus lateralis and two parts of the biceps femoris started in the late part of the swing phase and ended in the late period of the stance phase or the early period of the next swing phase. The semitendinosus showed two bursts of activity. The tensor fasciae latae acted when the previous muscles were inactive. Activity of the extensor digitorum longus was of low level but lasted during most of the step cycle. Correlation between electromyography, kinematics and anatomy helps us to understand the complex role of biarticular muscles: the tensor fasciae latae flexes the hip joint during the swing phase and extends the stifle joint during the stance phase, whereas the semitendinosus extends the hip joint during the stance phase and flexes the stifle joint during the swing phase. Cranial and caudal regions of the biceps femoris were also found to be bifunctional. Surface electromyography correlated with kinematic analysis gives valuable information about the functions and the timing of activity of the hindlimb muscles.


Naturally occurring BRCA2 alternative mRNA splicing events in clinically relevant samples.
BRCA1 and BRCA2 are the two principal tumour suppressor genes associated with inherited high risk of breast and ovarian cancer. Genetic testing of BRCA1/2 will often reveal one or more sequence variants of uncertain clinical significance, some of which may affect normal splicing patterns and thereby disrupt gene function. mRNA analyses are therefore among the tests used to interpret the clinical significance of some genetic variants. However, these could be confounded by the appearance of naturally occurring alternative transcripts unrelated to germline sequence variation or defects in gene function. To understand which novel splicing events are associated with splicing mutations and which are part of the normal BRCA2 splicing repertoire, a study was undertaken by members of the Evidence-based Network for the Interpretation of Germline Mutant Alleles (ENIGMA) consortium to characterise the spectrum of naturally occurring BRCA2 mRNA alternate-splicing events. mRNA was prepared from several blood and breast tissue-derived cells and cell lines by contributing ENIGMA laboratories. cDNA representing BRCA2 alternate splice sites was amplified and visualised using capillary or agarose gel electrophoresis, followed by sequencing. We demonstrate the existence of 24 different BRCA2 mRNA alternate-splicing events in lymphoblastoid cell lines and both breast cancer and non-cancerous breast cell lines. These naturally occurring alternate-splicing events contribute to the array of cDNA fragments that may be seen in assays for mutation-associated splicing defects. Caution must be observed in assigning alternate-splicing events to potential splicing mutations.


Decreased central mu-opioid receptor availability in fibromyalgia.
The underlying neurophysiology of acute pain is fairly well characterized, whereas the central mechanisms operative in chronic pain states are less well understood. Fibromyalgia (FM), a common chronic pain condition characterized by widespread pain, is thought to originate largely from altered central neurotransmission. We compare a sample of 17 FM patients and 17 age- and sex-matched healthy controls, using mu-opioid receptor (MOR) positron emission tomography. We demonstrate that FM patients display reduced MOR binding potential (BP) within several regions known to play a role in pain modulation, including the nucleus accumbens, the amygdala, and the dorsal cingulate. MOR BP in the accumbens of FM patients was negatively correlated with affective pain ratings. Moreover, MOR BP throughout the cingulate and the striatum was also negatively correlated with the relative amount of affective pain (McGill, affective score/sensory score) within these patients. These findings indicate altered endogenous opioid analgesic activity in FM and suggest a possible reason for why exogenous opiates appear to have reduced efficacy in this population.


Association of angiotensin-converting enzyme inhibitor therapy and comorbidity in diabetes: results from the Vermont diabetes information system.
Angiotensin converting enzyme inhibitors (ACE inhibitors) reduce peripheral vascular resistance via blockage of angiotensin converting enzyme (ACE). ACE inhibitors are commonly used to treat congestive heart failure and high blood pressure, but other effects have been reported. In this study, we explored the association between ACE inhibitor therapy and the prevalence of comorbid conditions in adults with diabetes We surveyed 1003 adults with diabetes randomly selected from community practices. Patients were interviewed at home and self-reported their personal and clinical characteristics including comorbidity. Current medications were obtained by direct observation of medication containers. We built logistic regression models with the history of comorbidities as the outcome variable and the current use of ACE inhibitors as the primary predictor variable. We adjusted for possible confounding by social (age, sex, alcohol drinking, cigarette smoking) and clinical factors (systolic blood pressure, body mass index (BMI), glycosolated hemoglobin (A1C), number of comorbid conditions, and number of prescription medications). ACE users reported a history of any cancer (except the non-life-threatening skin cancers) less frequently than non-users (10% vs. 15%; odd ratio = 0.59; 95% confidence interval [0.39, 0.89]; P = 0.01); and a history of stomach ulcers or peptic ulcer disease less frequently than non-users (12% vs. 16%, odd ratio = 0.70, [0.49, 1.01], P = 0.06). After correcting for potential confounders, ACE inhibitors remained significantly inversely associated with a personal history of cancer (odds ratio = 0.59, [0.39, 0.89]; P = 0.01) and peptic ulcer disease (odd ratio = 0.68, [0.46, 1.00], P = 0.05). ACE inhibitor use is associated with a lower likelihood of a history of cancer and peptic ulcers in patients with diabetes. These findings are limited by the cross sectional study design, self-report of comorbid diagnoses, and lack of information on the timing and duration of ACE inhibitor use. Further research is needed to confirm these associations and understand their mechanisms.


Ease of imagination, message framing, and physical activity messages.
The purpose of this research was to replicate a study that examined how message framing and ease of imagination interact to influence attitudes towards the prevention of heart disease through physical activity and a healthy diet. Changes were made such that only physical activity behaviour was profiled and assessed as a moderating variable. It was hypothesized that gain-framed messages would positively influence attitudes with hard to imagine symptoms, that loss-framed messages would positively influence attitudes with easy to imagine symptoms and exercise frequency would moderate the findings. This study employed a 2 (easy or hard to imagine symptoms) by 2 (gain- or loss-framed) Solomon square design whereby participants, half of whom completed a pre-test, were randomly assigned to one of four conditions: easy to imagine/gain-framed, hard to imagine/gain-framed, easy to imagine/loss-framed, or hard to imagine/loss-framed. Participants included adults over the age of 55 years (N=57) and undergraduate students (18-22 years; N=118). They were described either hard to imagine or easy to imagine symptoms of heart disease and diabetes and asked to imagine them. Participants then read either a gain- or loss-framed physical activity message followed by post-test questionnaires that assessed attitudes, exercise frequency, and demographics. Regression analyses showed no significant framing effects but significant effects for ease of imagination and exercise frequency as a moderating variable. This study failed to replicate the original research findings but showed that participants who exercised the least and were in the hard to imagine condition had the worst attitudes towards physical activity.


2-Aminopyrimidines as dual adenosine A1/A2A antagonists.
In this study thirteen 2-aminopyrimidine derivatives were synthesised and screened as potential antagonists of adenosine A1 and A2A receptors in order to further investigate the structure activity relationships of this class of compounds. 4-(5-Methylfuran-2-yl)-6-[3-(piperidine-1-carbonyl)phenyl]pyrimidin-2-amine (8m) was identified as a compound with high affinities for both receptors, with an A2AKi value of 6.34 nM and an A1Ki value of 9.54 nM. The effect of selected compounds on the viability of cultured cells was assessed and preliminary results indicate low cytotoxicity. In vivo efficacy at A2A receptors was illustrated for compounds 8k and 8m since these compounds attenuated haloperidol-induced catalepsy in rats. A molecular docking study revealed that the interactions between the synthesised compounds and the adenosine A2A binding site most likely involve Phe168 and Asn253, interactions which are similar for structurally related adenosine A2A receptor antagonists.


Streptococcus oralis subsp. dentisani Produces Monolateral Serine-Rich Repeat Protein Fibrils, One of Which Contributes to Saliva Binding via Sialic Acid.
Our studies reveal that the oral colonizer and cause of infective endocarditis Streptococcus oralis subsp. dentisani displays a striking monolateral distribution of surface fibrils. Furthermore, our data suggest that these fibrils impact the structure of adherent bacterial chains. Mutagenesis studies indicate that these fibrils are dependent on three serine-rich repeat proteins (SRRPs), here named fibril-associated protein A (FapA), FapB, and FapC, and that each SRRP forms a different fibril with a distinct distribution. SRRPs are a family of bacterial adhesins that have diverse roles in adhesion and that can bind to different receptors through modular nonrepeat region domains. Amino acid sequence and predicted structural similarity searches using the nonrepeat regions suggested that FapA may contribute to interspecies interactions, that FapA and FapB may contribute to intraspecies interactions, and that FapC may contribute to sialic acid binding. We demonstrate that a fapC mutant was significantly reduced in binding to saliva. We confirmed a role for FapC in sialic acid binding by demonstrating that the parental strain was significantly reduced in adhesion upon addition of a recombinantly expressed, sialic acid-specific, carbohydrate binding module, while the fapC mutant was not reduced. However, mutation of a residue previously shown to be essential for sialic acid binding did not decrease bacterial adhesion, leaving the precise mechanism of FapC-mediated adhesion to sialic acid to be defined. We also demonstrate that the presence of any one of the SRRPs is sufficient for efficient biofilm formation. Similar structures were observed on all infective endocarditis isolates examined, suggesting that this distribution is a conserved feature of this S. oralis subspecies.


Endovascular treatment of an internal carotid artery pseudoaneurysm following transsphenoidal surgery. Case report.
Internal carotid artery (ICA) pseudoaneurysm formation following transsphenoidal surgery is a rare but potentially lethal complication. Direct surgical repair with preservation of the ICA may be difficult. The feasibility of endovascular coil embolization with parent artery preservation for an iatrogenic ICA pseudoaneurysm is undefined. A 40-year-old man was referred to the authors' institution after identification of a pseudoaneurysm of the left ICA following transsphenoidal resection of a pituitary macroadenoma. The pseudoaneurysm was treated via an endovascular approach that included stent-assisted coil embolization of the lesion. Follow-up angiographic studies obtained 1 year later demonstrated complete occlusion of the aneurysm, and the patient remains asymptomatic. Stent-assisted coil embolization of this iatrogenic pseudoaneurysm was successful in achieving complete, angiographically confirmed aneurysm obliteration, with preservation of the ICA and short-term prevention of hemorrhage or carotidcavernous fistula. The endovascular method provided an effective, relatively low-risk treatment for this difficult lesion, and was an excellent alternative to direct surgical repair. Nonetheless, long-term follow-up review is required before definitive treatment recommendations can be made.


Defining the Most Appropriate Delivery Mode in Women with Inflammatory Bowel Disease: A Systematic Review.
High cesarean section (CS) rates are observed in patients with inflammatory bowel disease (IBD), but limited data are available to support this decision. We conducted a comprehensive review to evaluate the most appropriate mode of delivery in women with IBD according to disease phenotype and activity, as well as surgical history. We searched MEDLINE (source PubMed) and international conference abstracts, and included all studies that evaluated digestive outcome after delivery in patients with IBD. A total of 41 articles or abstracts were screened, and 18 studies were considered in this review, with sample sizes ranging from 4 to 229 patients and follow-up ranging from 2 months to 7.7 years. Pooled CS rates in patients without Perianal Crohn's disease (PCD), healed PCD or active PCD, were 27%, 43%, and 46%, respectively. Regarding the median rate of new PCD (3.0% [IQR, 1.5-11.5] versus 6.5% [0-19.7]) or PCD recurrence (13.5% [3.2-32.7] versus 45% [0-58]), no increase was observed in patients with vaginal delivery compared to CS, but for patients with an active disease, worsening of symptoms was noted in two-thirds of cases. Episiotomy, perianal tears, and instrumental delivery did not influence the incidence of PCD. In patients with ileal pouch anal anastomosis, uncomplicated vaginal delivery seemed to moderately influence pouch function, with no significant difference in terms of overall continence, daytime, or night-time stool frequency, or incontinence. However, these parameters seemed negatively impacted by a complicated vaginal delivery. New long-term data from well-designed studies are needed, but our review suggests that systematic CS in patients suffering from IBD should probably be limited to women at risk of perineal tears and obstetric injuries, with an active PCD, or with ileal pouch anal anastomosis.


Exposure to ionizing radiation and development of bone sarcoma: new insights based on atomic-bomb survivors of Hiroshima and Nagasaki.
Radiation-induced bone sarcoma has been associated with high doses of ionizing radiation from therapeutic or occupation-related exposures. However, the development of bone sarcoma following exposure to lower doses of ionizing radiation remains speculative. A cohort analysis based on the Life Span Study (n = 120,321) was performed to assess the development of bone sarcoma in atomic-bomb survivors of Hiroshima and Nagasaki followed from 1958 to 2001. The excess relative risk per gray of ionizing radiation absorbed by the bone marrow was estimated. Additional subject demographic, survival, and clinical factors were evaluated. Nineteen cases of bone sarcoma (in eleven males and eight females) were identified among the 80,181 subjects who met the inclusion criteria, corresponding to an incidence of 0.9 per 100,000 person-years. The mean ages at the time of the bombing and at diagnosis were 32.4 and 61.6 years, respectively. The mean bone marrow dose was 0.43 Gy. Osteosarcoma was the most commonly identified bone sarcoma. The most common bone sarcoma site was the pelvis. The overall unadjusted five-year survival rate was 25%. A dose threshold was found at 0.85 Gy (95% confidence interval, 0.12 to 1.85 Gy), with a linear dose-response association above this threshold. The linear slope equaled an excess relative risk of 7.5 per Gy (95% confidence interval, 1.34 to 23.14 per Gy) in excess of 0.85 Gy. On the basis of what we believe is one of the longest and largest prospective studies assessing the development of bone sarcoma in individuals exposed to ionizing radiation, it appears that the development of radiation-induced bone sarcoma may be associated with exposure to much lower doses of ionizing radiation than have previously been reported. Such new insights may potentially improve bone sarcoma prevention measures and broaden our understanding of the role of ionizing radiation from various sources on the development of malignant tumors. This study stresses the need to become increasingly aware of the various health risks that may be attributable to even low levels of ionizing radiation exposure. Prognostic Level I. See Instructions to Authors for a complete description of levels of evidence.


Extended uniportal video-assisted thoracic surgery for lung cancer: is it feasible?
Since the first description of uniportal video-assisted thoracic surgery (U-VATS) (or single-port) lobectomy, several centers in Asia and Europe rapidly adopted this technique as a standard approach for treatment of early stage non-small cell lung cancer (NSCLC). Despite the controversies regarding feasibility and completeness of resection, thoracic surgeons in high volume centers keep pushing the limits to perform very complex procedures also known as "extended resections" through minimally invasive surgery. Published series and case reports confirm the viability of U-VATS in highly complex surgical cases such as pneumonectomy, chest wall resection and bronchoplasty, which require experience and technical ability to be performed through a 3-6 cm single incision. In this article, the authors would like to present several clinical indications of locally advanced NSCLC and the technical aspects to accomplish an extended resection through U-VATS.


Germ line stem cell differentiation in Drosophila requires gap junctions and proceeds via an intermediate state.
Gap junctions coordinate processes ranging from muscle contraction to ovarian follicle development. Here we show that the gap junction protein Zero population growth (Zpg) is required for germ cell differentiation in the Drosophila ovary. In the absence of Zpg the stem cell daughter destined to differentiate dies. The zpg phenotype is novel, and we used this phenotype to genetically dissect the process of stem cell maintenance and differentiation. Our findings suggest that germ line stem cells differentiate upon losing contact with their niche, that gap junction mediated cell-cell interactions are required for germ cell differentiation, and that in Drosophila germ line stem cell differentiation to a cystoblast is gradual.


Photorhabdus adhesion modification protein (Pam) binds extracellular polysaccharide and alters bacterial attachment.
Photorhabdus are Gram-negative nematode-symbiotic and insect-pathogenic bacteria. The species Photorhabdus asymbiotica is able to infect humans as well as insects. We investigated the secreted proteome of a clinical isolate of P. asymbiotica at different temperatures in order to identify proteins relevant to the infection of the two different hosts. A comparison of the proteins secreted by a clinical isolate of P. asymbiotica at simulated insect (28 degrees C) and human (37 degrees C) temperatures led to the identification of a small and highly abundant protein, designated Pam, that is only secreted at the lower temperature. The pam gene is present in all Photorhabdus strains tested and shows a high level of conservation across the whole genus, suggesting it is both ancestral to the genus and probably important to the biology of the bacterium. The Pam protein shows limited sequence similarity to the 13.6 kDa component of a binary toxin of Bacillus thuringiensis. Nevertheless, injection or feeding of heterologously produced Pam showed no insecticidal activity to either Galleria mellonella or Manduca sexta larvae. In bacterial colonies, Pam is associated with an extracellular polysaccharide (EPS)-like matrix, and modifies the ability of wild-type cells to attach to an artificial surface. Interestingly, Surface Plasmon Resonance (SPR) binding studies revealed that the Pam protein itself has adhesive properties. Although Pam is produced throughout insect infection, genetic knockout does not affect either insect virulence or the ability of P. luminescens to form a symbiotic association with its host nematode, Heterorhabditis bacteriophora. We studied a highly abundant protein, Pam, which is secreted in a temperature-dependent manner in P. asymbiotica. Our findings indicate that Pam plays an important role in enhancing surface attachment in insect blood. Its association with exopolysaccharide suggests it may exert its effect through mediation of EPS properties. Despite its abundance and conservation in the genus, we find no evidence for a role of Pam in either virulence or symbiosis.


Safety of measles-containing vaccines in 1-year-old children.
All measles-containing vaccines are associated with several types of adverse events, including seizure, fever, and immune thrombocytopenia purpura (ITP). Because the measles-mumps-rubella-varicella (MMRV) vaccine compared with the separate measles-mumps-rubella (MMR) and varicella (MMR + V) vaccine increases a toddler's risk for febrile seizures, we investigated whether MMRV is riskier than MMR + V and whether either vaccine elevates the risk for additional safety outcomes. Study children were aged 12 to 23 months in the Vaccine Safety Datalink from 2000 to 2012. Nine study outcomes were investigated: 7 main outcomes (anaphylaxis, ITP, ataxia, arthritis, meningitis/encephalitis, acute disseminated encephalomyelitis, and Kawasaki disease), seizure, and fever. Comparing MMRV with MMR + V, relative risk was estimated by using stratified exact binomial tests. Secondary analyses examined post-MMRV or MMR + V risk versus comparison intervals; risk and comparison intervals were then contrasted for MMRV versus MMR+V. We evaluated 123,200 MMRV and 584,987 MMR + V doses. Comparing MMRV with MMR + V, risks for the 7 main outcomes were not significantly different. Several outcomes had few or zero postvaccination events. Comparing risk versus comparison intervals, ITP risk was higher after MMRV (odds ratio [OR]: 11.3 [95% confidence interval (CI): 1.9 to 68.2]) and MMR + V (OR: 10 [95% CI: 4.5 to 22.5]) and ataxia risk was lower after both vaccines (MMRV OR: 0.8 [95% CI: 0.5 to 1]; MMR + V OR: 0.8 [95% CI: 0.7 to 0.9]). Compared with MMR + V, MMRV increased risk of seizure and fever 7 to 10 days after vaccination. This study did not identify any new safety concerns comparing MMRV with MMR + V or after either the MMRV or the MMR + V vaccine. This study provides reassurance that these outcomes are unlikely after either vaccine.


Targeting of α7 Nicotinic Acetylcholine Receptors in the Treatment of Schizophrenia and the Use of Auditory Sensory Gating as a Translational Biomarker.
Accumulating evidence suggests that the α7 subtype of nicotinic acetylcholine receptors (nAChRs) plays a key role in inflammatory processes, thought to be involved in the pathophysiology of neuropsychiatric diseases, such as schizophrenia and Alzheimer's disease. Preclinical and clinical studies showed that the diminished suppression of P50 auditory evoked potentials in patients with schizophrenia may be associated with a decreased density of α7 nAChRs in the brain. This points to a role for auditory sensory gating (P50) as a translational biomarker. A number of agonists and positive allosteric modulators (PAMs) for α7 nAChR promoted beneficial effects in animal models with sensory gating and cognitive deficits. Additionally, several clinical studies showed that α7 nAChR agonists could improve suppression in auditory P50 evoked potentials, as well as cognitive deficits, and negative symptoms in patients with schizophrenia. Taken together, α7 nAChR presents as an extremely attractive therapeutic target for schizophrenia. In this article, the author discusses recent findings on α7 nAChR agonists such as DMXB-A, RG3487, TC-5619, tropisetron, EVP-6124 (encenicline), ABT-126, AQW051 and α7 nAChR PAMs such as JNJ-39393406, PNU- 120596 and AVL-3288 (also known as UCI-4083), and their potential as therapeutic drugs for neuropsychiatric diseases, such as schizophrenia.


Starches, sugars and obesity.
The rising prevalence of obesity, not only in adults but also in children and adolescents, is one of the most important public health problems in developed and developing countries. As one possible way to tackle obesity, a great interest has been stimulated in understanding the relationship between different types of dietary carbohydrate and appetite regulation, body weight and body composition. The present article reviews the conclusions from recent reviews and meta-analyses on the effects of different starches and sugars on body weight management and metabolic disturbances, and provides an update of the most recent studies on this topic. From the literature reviewed in this paper, potential beneficial effects of intake of starchy foods, especially those containing slowly-digestible and resistant starches, and potential detrimental effects of high intakes of fructose become apparent. This supports the intake of whole grains, legumes and vegetables, which contain more appropriate sources of carbohydrates associated with reduced risk of cardiovascular and other chronic diseases, rather than foods rich in sugars, especially in the form of sugar-sweetened beverages.


Medial trigonal arteriovenous malformations.
A retrospective analysis of 48 patients of medial paratrigonal arteriovenous malformations (AVMs) which constituted 18% of the total 258. AVMs surgically managed in our institute for 16 years was carried out to study their clinical presentation, radiological features, operative approaches and functional outcome. While hemorrhage was the initial presenting symptom in 35, it was longstanding headache in 10, focal seizures in two and progressive weakness in one. However, 41 of these presented with bleed at the time of admission to our hospital and in 15 there was history of multiple bleeds. Twelve and eleven patients had field cut and hemiparesis respectively. Arterial supply to the malformation was quite uniform with posterior (43 patients) and anterior cerebral (25 patients) being the most frequent source. Venous drainage was predominantly into the galenic system (70%). Preoperative embolization was carried out in six patients. While the AVM was excised through a parasagittal interhemispheric approach in 34 patients, the nidus was approached through a direct transcortical (superior parietal lobule) approach in fourteen patients. There was only one operative mortality. Twelve patients whose preoperative fields were normal developed field cut postoperatively. An improvement of the preoperative field deficit was noted only in one patient. While eight of the ten patients with preoperative weakness improved and remained independent the remaining two were moderately disabled. Only two of the 10 patients who developed postoperative weakness remained severely disabled. Our operative results suggest that these AVMs which are prone to recurrent bleeds can be resected with acceptable morbidity. Hemianopia resulting from bleed or surgery is unlikely to improve.


Anaplastic thyroid cancer: molecular pathogenesis and emerging therapies.
Anaplastic thyroid cancer (ATC) is a rare malignancy. While external beam radiation therapy has improved locoregional control, the median survival of approximately 4 months has not changed in more than half a century due to uncontrolled systemic metastases. The objective of this study was to review the literature in order to identify potential new strategies for treating this highly lethal cancer. PubMed searches were the principal source of articles reviewed. The molecular pathogenesis of ATC includes mutations in BRAF, RAS, catenin (cadherin-associated protein), beta 1, PIK3CA, TP53, AXIN1, PTEN, and APC genes, and chromosomal abnormalities are common. Several microarray studies have identified genes and pathways preferentially affected, and dysregulated microRNA profiles differ from differentiated thyroid cancers. Numerous proteins involving transcription factors, signaling pathways, mitosis, proliferation, cell cycle, apoptosis, adhesion, migration, epigenetics, and protein degradation are affected. A variety of agents have been successful in controlling ATC cell growth both in vitro and in nude mice xenografts. While many of these new compounds are in cancer clinical trials, there are few studies being conducted in ATC. With the recent increased knowledge of the many critical genes and proteins affected in ATC, and the extensive array of targeted therapies being developed for cancer patients, there are new opportunities to design clinical trials based upon tumor molecular profiling and preclinical studies of potentially synergistic combinatorial novel therapies.


Experimental Confirmation of the Universal Law for the Vibrational Density of States of Liquids.
An analytical model describing the vibrational density of states (VDOS) of liquids has long been elusive, owing to the complexities of liquid dynamics. Nevertheless, Zaccone and Baggioli have recently developed such a model which was proposed to be the universal law for the vibrational density of states of liquids. Distinct from the Debye law, g(ω) ∝ ω2, for solids, the universal law for liquids reveals a linear relationship, g(ω) ∝ ω, in the low-energy region. We have confirmed this universal law with experimental VDOS measured by inelastic neutron scattering on real liquid systems including water, liquid metal, and polymer liquids, and have applied this model to extract the effective relaxation rate for the short time dynamics for each liquid. The model has also been further evaluated in the prediction of the specific heat with comparison to existing experimental data as well as with values obtained by different approaches.


Spiritual Needs of Families With Bereavement and Loss of an Infant in the Neonatal Intensive Care Unit: A Qualitative Study.
The hospital is a place full of distress and questions about the meaning of life. The death of a child can cause a spiritual struggle and crisis. Therefore, it is necessary for health care providers in the neonatal intensive care unit (NICU) to assess the spiritual needs of families that have lost a child. The purpose of this study was to explore the spiritual needs of families in Iran at the end of their baby's life and through bereavement in the NICU. This study was an exploratory qualitative study performed using purposeful sampling and semi-structured interviews with 24 participants. Inclusion criteria for families, nurses, and physicians included having experienced at least one newborn death in the last six months in the NICU. The research environment was the NICU in Isfahan, one of the largest cities in Iran. Data analysis revealed three main themes: spiritual belief in a supernatural power, the need for comfort of the soul, and human dignity for the newborn. The results of this study created a new vision in addressing spiritual needs of Iranian families who experience the death of a newborn.


Super-Enhancers Promote Transcriptional Dysregulation in Nasopharyngeal Carcinoma.
Nasopharyngeal carcinoma (NPC) is an invasive cancer with particularly high incidence in Southeast Asia and Southern China. The pathogenic mechanisms of NPC, particularly those involving epigenetic dysregulation, remain largely elusive, hampering clinical management of this malignancy. To identify novel druggable targets, we carried out an unbiased high-throughput chemical screening and observed that NPC cells were highly sensitive to inhibitors of cyclin-dependent kinases (CDK), especially THZ1, a covalent inhibitor of CDK7. THZ1 demonstrated pronounced antineoplastic activities both in vitro and in vivo An integrative analysis using both whole-transcriptome sequencing and chromatin immunoprecipitation sequencing pinpointed oncogenic transcriptional amplification mediated by super-enhancers (SE) as a key mechanism underlying the vulnerability of NPC cells to THZ1 treatment. Further characterization of SE-mediated networks identified many novel SE-associated oncogenic transcripts, such as BCAR1, F3, LDLR, TBC1D2, and the long noncoding RNA TP53TG1. These transcripts were highly and specifically expressed in NPC and functionally promoted NPC malignant phenotypes. Moreover, DNA-binding motif analysis within the SE segments suggest that several transcription factors (including ETS2, MAFK, and TEAD1) may help establish and maintain SE activity across the genome. Taken together, our data establish the landscape of SE-associated oncogenic transcriptional network in NPC, which can be exploited for the development of more effective therapeutic regimens for this disease. Cancer Res; 77(23); 6614-26. ©2017 AACR.


Performance-limiting factors for GaAs-based single nanowire photovoltaics.
GaAs nanowires (NWs) offer the possibility of decoupling light absorption from charge transport for high-performance photovoltaic (PV) devices. However, it is still an open question as to whether these devices can exceed the Shockley-Queisser efficiency limit for single-junction PV. In this work, single standing GaAs-based nanowire solar cells in both radial and vertical junction configurations is analyzed and compared to a planar thin-film design. By using a self-consistent, electrical-optically coupled 3D simulator, we show the design principles for nanowire and planar solar cells are significantly different; nanowire solar cells are vulnerable to surface and contact recombination, while planar solar cells suffer significant losses due to imperfect backside mirror reflection. Overall, the ultimate efficiency of the GaAs nanowire solar cell with radial and vertical junction is not expected to exceed that of the thin-film design, with both staying below the Shockley-Queisser limit.


CXC Chemokines Exhibit Bactericidal Activity against Multidrug-Resistant Gram-Negative Pathogens.
The continued rise and spread of antimicrobial resistance among bacterial pathogens pose a serious challenge to global health. Countering antimicrobial-resistant pathogens requires a multifaceted effort that includes the discovery of novel therapeutic approaches. Here, we establish the capacity of the human CXC chemokines CXCL9 and CXCL10 to kill multidrug-resistant Gram-negative bacteria, including New Delhi metallo-beta-lactamase-1-producing Klebsiella pneumoniae and colistin-resistant members of the family Enterobacteriaceae that harbor the mobile colistin resistance protein MCR-1 and thus possess phosphoethanolamine-modified lipid A. Colistin-resistant K. pneumoniae isolates affected by genetic mutation of the PmrA/PmrB two-component system, a chromosomally encoded regulator of lipopolysaccharide modification, and containing 4-amino-4-deoxy-l-arabinose-modified lipid A were also found to be susceptible to chemokine-mediated antimicrobial activity. However, loss of PhoP/PhoQ autoregulatory control, caused by disruption of the gene encoding the negative regulator MgrB, limited the bactericidal effects of CXCL9 and CXCL10 in a variable, strain-specific manner. Cumulatively, these findings provide mechanistic insight into chemokine-mediated antimicrobial activity, highlight disparities amongst determinants of colistin resistance, and suggest that chemokine-mediated bactericidal effects merit additional investigation as a therapeutic avenue for treating infections caused by multidrug-resistant pathogens.IMPORTANCE As bacterial pathogens become resistant to multiple antibiotics, the infections they cause become increasingly difficult to treat. Carbapenem antibiotics provide an essential clinical barrier against multidrug-resistant bacteria; however, the dissemination of bacterial enzymes capable of inactivating carbapenems threatens the utility of these important antibiotics. Compounding this concern is the global spread of bacteria invulnerable to colistin, a polymyxin antibiotic considered to be a last line of defense against carbapenem-resistant pathogens. As the effectiveness of existing antibiotics erodes, it is critical to develop innovative antimicrobial therapies. To this end, we demonstrate that the chemokines CXCL9 and CXCL10 kill the most concerning carbapenem- and colistin-resistant pathogens. Our findings provide a unique and timely foundation for therapeutic strategies capable of countering antibiotic-resistant "superbugs."


General practitioners' referrals to specialist outpatient clinics. I. Why general practitioners refer patients to specialist outpatient clinics.
There has been much concern about the wide variations in general practitioners' referral rates and the consequent implications for cost and quality of care. This has led to a call to evaluate the appropriateness and effectiveness of referrals. A collaborative audit of referrals to outpatient clinics was conducted by 127 general practitioners in 33 practices in the Oxford region. Records were kept of 18,754 referrals, which included data on diagnoses and reasons for referral. Overall, 6553 (35.4%) of the referrals were for particular treatments or operations and a further 6475 (34.9%) were for specific investigation or diagnosis. Advice on management was the main reason for referral in 2656 (14.3%) cases, and in 1719 (9.3%) cases the general practitioners wanted the consultants to take over managing their patients. Reassurance of either the general practitioner or the patient was recorded as the main reason in only 762 (4.1%) referrals. There seems to be scope for rationalising the referral process. A programme with three stages for evaluating referrals to outpatient clinics is recommended.


ECOTYPIC DIVERGENCE IN ALPINE POLEMONIUM VISCOSUM: GENETIC STRUCTURE, QUANTITATIVE VARIATION, AND LOCAL ADAPTATION.
Polemonium viscosum has a continuous distribution from 3,500 m in the krummholz to 4,025 m on the summit ridges of Pennsylvania Mountain, Colorado. Seeds produced by plants at opposite ends of this cline, 1.5 km apart, differed significantly at allozyme loci in two consecutive breeding seasons. Mean multilocus Fst values for both years (0.015 and 0.069) were significantly different from zero, indicating restricted gene exchange between subpopulations. Average allele frequencies at two individual loci also differed significantly between families comprising krummholz and summit subpopulations. Progeny of plants growing on the summit had higher leaf production rates, more densely packed leaflets, and lower resistance to aphids than progeny of plants growing in the krummholz site, when tested under greenhouse conditions. These differences probably reflect the restricted opportunities for growth and severe exposure at high elevations, and the increased risk from herbivores near timberline. The two subpopulations did not differ in leaf length (stature), leaf width, or pubescence. Reciprocal transplanting of seedlings between krummholz and summit sites confirmed that the differences were adaptive, since progeny from each subpopulation performed significantly better in their parent's habitat. Coordinated studies of genetic structure, quantitative variation, and local adaptation across the elevational range of P. viscosum provide a comprehensive view of ecotypic divergence in this widespread alpine plant.


An Exploration of Edible Palm Weevil Larvae (Akokono) as a Source of Nutrition and Livelihood: Perspectives From Ghanaian Stakeholders.
Meeting the nutritive needs of infants and young children is a challenge in Ghana. Alternative animal source foods, including insects, could enhance infant and young child dietary quality while also improving livelihoods. To investigate the perspectives of Ghanaian stakeholders on the acceptability of the palm weevil larvae ( akokono) as a food source and the feasibility of micro-farming this local edible insect as a complementary food for infants and young children. We conducted an ethnographic study in the Brong-Ahafo Region of Ghana. First, 48 caregivers were asked questions about feeding and care practices, including attitudes toward production and consumption of akokono. Then, a selection of previously interviewed respondents joined 1 of 8 focus group discussions to provide further insight on key themes that emerged from earlier interviews. Concurrently, interviews with 25 other key local stakeholders were conducted. Respondents generally had favorable perceptions of akokono as a nutritious food. A small minority would not consume akokono for religious reasons. Key factors positively influencing the acceptability of akokono as a complementary food were familiarity with the consumption of akokono by the primary caregiver and health worker endorsement of akokono. Stakeholders consider the larvae farmable and were open to its domestication. Anticipated barriers to scaling up akokono micro-farming include a need for greater familiarity with and acceptance of the insect as food for infants and young children and creation of a sustainable market. Engagement with stakeholders, including health workers, will facilitate use of akokono as a complementary food.


Simple Aesthetic Sense and Addiction Emerge in Neural Relations of Cost-Benefit Decision in Foraging.
A rudimentary aesthetic sense is found in the stimulus valuations and cost-benefit decisions made by primitive generalist foragers. These are based on factors governing personal economic decisions: incentive, appetite, and learning. We find that the addictive process is an extreme expression of aesthetic dynamics. An interactive, agent-based model, ASIMOV, reproduces a simple aesthetic sense from known neural relations of cost-benefit decision in foraging. In the presence of very high reward, an addiction-like process emerges. A drug-like prey provides extreme reward with no nutritive value, initiating high selectivity and prolonged cravings for drug through reward learning. Varying reward experience, caused by homeostatic changes in the neural circuitry of reward, further establishes the course of addiction, consisting of desensitization, withdrawal, resensitization, and associated changes in nutritional choice and pain sensitivity. These observations are consistent with the early evolution of addiction mechanisms in simple generalist foragers as an aesthetic sense for evaluating prey. ASIMOV is accessible to inspection, modification, and experiment, is adaptable as an educational tool, and provides insight on the possible coevolutionary origins of aesthetics and the addiction process.


Endoscopic findings predict the histologic diagnosis in gastrointestinal graft-versus-host disease.
Graft-versus-host disease (GvHD) of the gastrointestinal tract is a major cause of morbidity and mortality after allogeneic bone marrow transplantation (BMT). Whether endoscopic findings predict the histologic diagnosis of GvHD in the gastrointestinal tract remains controversial. We performed a study to determine the diagnostic accuracy of macroscopic endoscopy findings in the diagnosis of acute and chronic histologically proven gastrointestinal GvHD (GI-GvHD). Endoscopic images from the intestinal mucosa of post-BMT patients were blindly graded as positive or negative for GI-GvHD and compared with corresponding histological findings, which were used as the gold standard. 44 BMT patients were referred for 96 endoscopic evaluations. Using 162 endoscopy-biopsy pairs, a positive association between endoscopic grading and histologic grading of GI-GvHD (odds ratio [OR] = 11.97, 95% CI 3.86, 37.16) was observed. Endoscopic diagnosis correctly predicted histologic diagnosis in both acute and chronic GI-GvHD (OR = 9.3 vs. 23.1, P = 0.31). The diagnostic accuracy of endoscopy was high in both acute and chronic histologically proven GI-GvHD. Accurate diagnosis of GI-GvHD might be obtained with mucosal biopsies from either the upper or lower gastrointestinal tract. Endoscopy may play a significant role in establishing early diagnosis and treatment for GI-GvHD in patients following BMT, but histologic evaluation of the gastrointestinal mucosa is needed to confirm the final diagnosis.


Metabarcoding to investigate changes in soil microbial communities within forensic burial contexts.
The estimation of the time elapsed since death (post-mortem interval, or PMI) is one of the key themes that forensic scientists have to address frequently. However, the estimation of PMI still suffers from poor accuracy and biases especially when decomposition stages are prolonged, so further improvements in methods for PMI estimation are desirable. Soil microbial communities associated with decomposing bodies have been shown to be good candidates for the estimation of the PMI of exposed bodies. Nevertheless, further research is required to better understand the bacterial succession associated with decomposition of buried carcasses in order to test its reliability and applicability for the estimation of PMI and to better understand the dynamics involved with decomposition within this particular scenario. Therefore we explored the succession of soil microbial communities associated with four decomposing pig carcasses (from one to six months PMI) using a metabarcoding approach. The sequencing of the bacterial 16S rRNA variable region 4 (V4) revealed trends linking particular microbial taxa with specific PMIs, and notably an increase in Proteobacteria, Firmicutes and Bacteroidetes at specific PMIs as well as a decrease in Acidobacteria. Our results, in accordance with previous studies conducted on exposed bodies of different mammalian species (including humans), also showed a general reduction of the taxonomic richness from two months PMI onwards, as well as an incomplete re-establishment of the starting soil microbial conditions after six months PMI. We also found specific mammal-derived taxa, such as Bacteroides spp., being still present in the soil after six months PMI. As such, this study serves as a baseline for additional research to allow the characterisation of biomarkers associated with specific PMIs. Due to the similarity between the results presented here and those reported in other types of decomposition studies we believe that the metabarcoding approach has considerable potential in the estimation of the PMI, particularly to clarify cases involving heavily skeletonised bodies or for the investigation of clandestine graves in which the carcass has been moved from its original place of deposition.


Deregulation of the p57-E2F1-p53 axis results in nonobstructive hydrocephalus and cerebellar malformation in mice.
The cyclin-dependent kinase inhibitor (CKI) p57(Kip2) plays a pivotal role in cell cycle arrest during development, in particular, in the regulation of the entry of proliferating progenitors into quiescence. The gene encoding p57 undergoes genomic imprinting, and impairment of the regulation of p57 expression results in various developmental anomalies in humans and mice. We now show that p57 is expressed predominantly in the subcommissural organ and cerebellar interneurons in the mouse brain and that mice with brain-specific deletion of the p57 gene (Kip2) manifest prominent nonobstructive hydrocephalus as well as cerebellar malformation associated with the loss of Pax2-positive interneuron precursors and their descendants, including Golgi cells and γ-aminobutyric acid-containing neurons of the deep cerebellar nuclei. These abnormalities were found to be attributable to massive apoptosis of precursor cells in the developing brain. The morphological defects of the p57-deficient mice were corrected by knock-in of the gene for the related CKI p27(Kip1) at the Kip2 locus. The abnormalities were also prevented by additional genetic ablation of p53 or E2F1. Our results thus implicate p57 in cell cycle arrest in the subcommissural organ and Pax2-positive interneuron precursors, with the lack of p57 resulting in induction of p53-dependent apoptosis due to hyperactivation of E2F1.


BCG and Adverse Events in the Context of Leprosy.
Notwithstanding its beneficial immunoprophylactic outcomes regarding leprosy and childhood TB, BCG vaccination may cause adverse events, particularly of the skin. However, this local hyper-immune reactivity cannot be predicted before vaccination, nor is its association with protection against leprosy known. In this study we investigated the occurrence of adverse events after BCG (re)vaccination in contacts of leprosy patients and analyzed whether the concomitant systemic anti-mycobacterial immunity was associated with these skin manifestations. Within a randomized controlled BCG vaccination trial in Bangladesh, 14,828 contacts of newly diagnosed leprosy patients received BCG vaccination between 2012 and 2017 and were examined for adverse events 8 to 12 weeks post-vaccination. From a selection of vaccinated contacts, venous blood was obtained at follow-up examination and stimulated with Mycobacterium leprae (M. leprae) antigens in overnight whole-blood assays (WBA). M. leprae phenolic glycolipid-I-specific antibodies and 32 cytokines were determined in WBAs of 13 individuals with and 13 individuals without adverse events after vaccination. Out of the 14,828 contacts who received BCG vaccination, 50 (0.34%) presented with adverse events, mainly (80%) consisting of skin ulcers. Based on the presence of BCG scars, 30 of these contacts (60%) had received BCG in this study as a booster vaccination. Similar to the pathological T-cell immunity observed for tuberculoid leprosy patients, contacts with adverse events at the site of BCG vaccination showed elevated IFN-γ levels in response to M. leprae-specific proteins in WBA. However, decreased levels of sCD40L in serum and GRO (CXCL1) in response to M. leprae simultaneously indicated less T-cell regulation in these individuals, potentially causing uncontrolled T-cell immunity damaging the skin. Skin complications after BCG vaccination present surrogate markers for protective immunity against leprosy, but also indicate a higher risk of developing tuberculoid leprosy. Netherlands Trial Register: NTR3087.


Accuracy of 6 routine 25-hydroxyvitamin D assays: influence of vitamin D binding protein concentration.
Recent recognition of its broad pathophysiological importance has triggered an increased interest in 25-hydroxyvitamin D [25(OH)D]. By consequence, throughput in 25(OH)D testing has become an issue for clinical laboratories, and several automated assays for measurement of 25(OH)D are now available. The aim of this study was to test the accuracy and robustness of these assays by comparing their results to those of an isotope dilution/online solid-phase extraction liquid chromatography/tandem mass spectrometry (ID-XLC-MS/MS) method. We put specific focus on the influence of vitamin D-binding protein (DBP) by using samples with various concentrations of DBP. We used 5 automated assays (Architect, Centaur, iSYS, Liaison, and Elecsys), 1 RIA (Diasorin) preceded by extraction, and an ID-XLC-MS/MS method to measure 25(OH)D concentrations in plasma samples of 51 healthy individuals, 52 pregnant women, 50 hemodialysis patients, and 50 intensive care patients. Using ELISA, we also measured DBP concentrations in these samples. Most of the examined 25(OH)D assays showed significant deviations in 25(OH)D concentrations from those of the ID-XLC-MS/MS method. As expected, DBP concentrations were higher in samples of pregnant women and lower in samples of IC patients compared to healthy controls. In 4 of the 5 fully automated 25(OH)D assays, we observed an inverse relationship between DBP concentrations and deviations from the ID-XLC-MS/MS results. 25(OH)D measurements performed with most immunoassays suffer from inaccuracies that are DBP concentration dependent. Therefore, when interpreting results of 25(OH)D measurements, careful consideration of the measurement method is necessary.


C1 esterase inhibitor activity in amniotic fluid embolism.
Amniotic fluid embolism exhibits activation of the complement system and the kallikrein-kinin and coagulofibrinolytic systems. C1 esterase inhibitor is a major inhibitor of C1 esterase and can inhibit plasma kallikrein and also factors XIIa and XIa. Its activity has been shown to be significantly lower in pregnancy and labor than in the nonpregnant state. The purpose of this study was to determine C1 esterase inhibitor activity levels in amniotic fluid embolism. Retrospective study. A single university-based center. One hundred six cases with amniotic fluid embolism in a total of 194 singleton pregnant women between January 2010 and December 2011. None. One hundred six cases of amniotic fluid embolism had applied to the Japan amniotic fluid embolism registration center in Hamamatsu University School of Medicine between January 2010 and December 2011. In amniotic fluid embolism cases, 85 cases were nonfatal and 21 cases were fatal. Eighty-eight women who delivered without amniotic fluid embolism were regarded as a control. C1 esterase inhibitor activity levels were significantly lower in amniotic fluid embolism patients (30.0% ± 1.8%) than in control women (62.0% ± 2.0%) (p < 0.0001). C1 esterase inhibitor activity levels in fatal amniotic fluid embolism cases (22.5% ± 3.4%) were significantly lower than those in nonfatal amniotic fluid embolism cases (32.0% ± 2.1%) (p < 0.05). These results demonstrated that low C1 esterase inhibitor activity levels were closely associated with the pathogenesis of amniotic fluid embolism suggesting that C1 esterase inhibitor activity levels have potential as a prognosis factor of amniotic fluid embolism.


Thermal properties of lauric acid filled in carbon nanotubes as shape-stabilized phase change materials.
Carbon nanotubes (CNTs) filled with lauric acid (LA) as a kind of shape-stabilized phase change material were prepared and their structures and phase change properties were characterized. The results showed that the melting point and latent heat of LA confined in carbon nanotubes were lower than those of the bulk material, and both decrease as the diameters of CNTs and the filling ratios of LA decrease. Molecular dynamics (MD) simulations indicated that LA molecules form a liquid layer near pore walls and crystallize at the pore center. When the LA filling ratio was reduced to a certain value, all LA molecules were attached to the inner walls of CNTs, hindering their crystallization. A linear relationship between the melting temperature shift and structural properties was obtained based on the modified Gibbs-Thomson equation, which gives a reliable interpretation of the size effect of nanochannels in phase change materials. We also found that the thermal conductivity of the composite CNTs/LA was four times larger than that of pure LA. This study will provide insights into the design of novel composite phase change materials with better thermal properties by the selection of suitable porous materials and tailoring their pore structures.


Modulation of short intra-cortical inhibition during action reprogramming.
Actions are selected in the context of environmental demands and internal goals. Since both change continuously it is often necessary to inhibit a prepared action plan in favour of an alternative, a process we refer to as action reprogramming. Previous studies have established that a frontal/basal ganglia network exerts top-down control over the primary motor cortex (M1) during action reprogramming. The current study focuses on the role of M1 itself during action reprogramming. Participants were asked to perform a behavioural task that required them to either execute a prepared response or to reprogram an alternative response. Paired-pulse TMS was used to investigate short-interval intra-cortical inhibition (SICI) during these action execution and action reprogramming trials. Normal action execution was associated with sustained SICI in the M1 during both trials in which the contralateral hand was to respond and trials in which the ipsilateral hand was to respond. In contrast, reprogramming towards an alternative action was associated with a progressive release of SICI in M1 involved in the execution of the novel response. This release started 125 ms after the cue telling the participants to reprogram their action. This time point is consistent with previous results showing a facilitatory influence of the pre-supplementary motor area (pre-SMA) on the M1 at the same delay. Hence, SICI might be a potential candidate mechanism through which frontal lobe areas could influence primary motor cortex output.


Development of a novel carboplatin like cytoplasmic trackable near infrared fluorophore conjugate via strain-promoted azide alkyne cycloaddition.
The successful design and pre-clicked synthesis of a non-toxic and cytosol trackable carboplatin-like near infrared fluorophore conjugate via strain-promoted azide alkyne cycloaddition (SPAAC) is reported. Reaction of cis-[Pt(2-azidopropane-1,3-diamine)(cbdca)] (cbdca = cyclobutane-1,1-dicarboxylato) and a bicyclo[6.1.0]non-4-yne near-infrared (NIR) azadipyrromethene fluorophore gave the corresponding clicked Pt-Fluorophore conjugate. The X-ray crystal structure of cis-[Pt(2-azidopropane-1,3-diamine)(cbdca)] was determined featuring the azide on the bidentate 1,3-diaminopropane ligand. The Pt-fluorophore conjugate is the first example of a Pt conjugate clicked via strain-promoted azide alkyne cycloaddition (SPAAC) where the reactive azide handle is on the amine carrier ligand. The in vitro cytotoxicity and widefield fluorescence imaging of the Pt-Fluorophore conjugate in A2780P and A2780cisR cells are described.


Is floral diversification associated with pollinator divergence? Flower shape, flower colour and pollinator preference in Chilean Mimulus.
Adaptation to different pollinators is thought to drive divergence in flower colour and morphology, and may lead to interspecific reproductive isolation. Floral diversity was tested for association with divergent pollinator preferences in a group of four closely related wildflower species: the yellow-flowered Mimulus luteus var. luteus and the red-pigmented M. l. variegatus, M. naiandinus and M. cupreus. Patterns of pollinator visitation were evaluated in natural plant populations in central Chile, including both single-species and mixed-species sites. Floral anthocyanin pigments were identified, and floral morphology and nectar variation were quantified in a common garden experiment using seeds collected from the study sites. Mimulus l. luteus, M. l. variegatus and M. naiandinus are morphologically similar and share a single generalist bumblebee pollinator, Bombus dahlbomii. Mimulus cupreus differs significantly from the first three taxa in corolla shape as well as nectar characteristics, and had far fewer pollinator visits. This system shows limited potential for pollinator-mediated restriction of gene flow as a function of flower colour, and no evidence of transition to a novel pollinator. Mimulus cupreus may experience reduced interspecific gene flow due to a lack of bumblebee visitation, but not because of its red pigmentation: rare yellow morphs are equally undervisited by pollinators. Overall, the results suggest that factors other than pollinator shifts may contribute to the maintenance of floral diversity in these Chilean Mimulus species.


Appropriate Use of Short-Course Antibiotics in Common Infections: Best Practice Advice From the American College of Physicians.
Antimicrobial overuse is a major health care issue that contributes to antibiotic resistance. Such overuse includes unnecessarily long durations of antibiotic therapy in patients with common bacterial infections, such as acute bronchitis with chronic obstructive pulmonary disease (COPD) exacerbation, community-acquired pneumonia (CAP), urinary tract infections (UTIs), and cellulitis. This article describes best practices for prescribing appropriate and short-duration antibiotic therapy for patients presenting with these infections. The authors conducted a narrative literature review of published clinical guidelines, systematic reviews, and individual studies that addressed bronchitis with COPD exacerbations, CAP, UTIs, and cellulitis. This article is based on the best available evidence but was not a formal systematic review. Guidance was prioritized to the highest available level of synthesized evidence. Clinicians should limit antibiotic treatment duration to 5 days when managing patients with COPD exacerbations and acute uncomplicated bronchitis who have clinical signs of a bacterial infection (presence of increased sputum purulence in addition to increased dyspnea, and/or increased sputum volume). Clinicians should prescribe antibiotics for community-acquired pneumonia for a minimum of 5 days. Extension of therapy after 5 days of antibiotics should be guided by validated measures of clinical stability, which include resolution of vital sign abnormalities, ability to eat, and normal mentation. In women with uncomplicated bacterial cystitis, clinicians should prescribe short-course antibiotics with either nitrofurantoin for 5 days, trimethoprim-sulfamethoxazole (TMP-SMZ) for 3 days, or fosfomycin as a single dose. In men and women with uncomplicated pyelonephritis, clinicians should prescribe short-course therapy either with fluoroquinolones (5 to 7 days) or TMP-SMZ (14 days) based on antibiotic susceptibility. In patients with nonpurulent cellulitis, clinicians should use a 5- to 6-day course of antibiotics active against streptococci, particularly for patients able to self-monitor and who have close follow-up with primary care.


B-type natruiretic peptide levels stratify the risk for arrhythmia among implantable cardioverter defibrillator patients.
We sought to study the association between brain natriuretic peptide (BNP) levels and the occurrence of ventricular arrhythmias in patients with left ventricular dysfunction (LVD) and an implantable cardioverter defibrillator (ICD). This was a prospective study of consecutive, stable, ambulatory patients with moderate and severe ischemic LVD and an ICD. A plasma BNP level was obtained at recruitment. Patients were evaluated every 3-6 mo for a minimum of 1 y. The primary end point was the occurrence of malignant ventricular arrhythmia or sudden cardiac death (SCD). The cohort consisted of 94 subjects (6 women) with a mean +/- standard deviation (SD) age of 69 +/- 10 y. The ICD implantation indication was primary and secondary prevention of SCD in 49% and 51% of subjects, respectively. A primary end point occurred in 27 patients (29%), and was more frequent in symptomatic heart failure patients and those implanted for secondary prevention of SCD. The median BNP level was significantly higher among patients who experienced an end point (191 pg/ml versus 142 pg/ml, p = 0.03). After controlling for New York Heart Association heart failure class and ICD implantation indication, the odds ratio (OR) for experiencing an adverse outcome among the upper BNP quartile versus all others was 3.5 (95% confidence interval [CI]: 1.2-10.2). Among patients implanted for primary prevention of SCD, none of the patients in the lower BNP quartile (BNP < 91 pg/ml) experienced an adverse outcome. These results suggest that abnormally high BNP levels not only predict cardiac death, but also arrhythmic death in this patient population, and a low BNP level can serve to identify low risk patients. (c)


Relationships within Cupressaceae sensu lato: a combined morphological and molecular approach.
Parsimony analysis of matK and rbcL sequence data, together with a nonmolecular database, yielded a well-resolved phylogeny of Cupressaceae sensu lato. Monophyly of Cupressaceae sensu stricto is well supported, and separate northern and southern hemisphere subclades are resolved, with Tetraclinis within the northern subclade; there is no support for any of the tribes sensu Li. Taxodiaceae comprise five separate lineages. Chamaecyparis nootkatensis falls within Cupressus, clustering with a robust clade of New World species. Libocedrus Florin is paraphyletic and should incorporate Pilgerodendron. Evolution of several characters of wood and leaf anatomy and chemistry is discussed in light of this estimate of the phylogeny; numerous parallelisms are apparent. A new infrafamilial classification is proposed in which seven subfamilies are recognized: Callitroideae Saxton, Athrotaxidoideae Quinn, Cunninghamioideae (Sieb. & Zucc.) Quinn, Cupressoideae Rich. ex Sweet, Sequoioideae (Luerss.) Quinn, Taiwanioideae (Hayata) Quinn, Taxodioideae Endl. ex K. Koch. The rbcL sequence for Taxodium distichum is corrected, and the implications for a previously published estimate of the minimum rate of divergence of the gene since the Miocene are highlighted.


Pharmacodynamics and arteriovenous difference of intravenous naloxone in healthy volunteers exposed to remifentanil.
Pharmacodynamic studies of naloxone require opioid agonism. Steady state condition may be achieved by remifentanil TCI (target controlled infusion). Opioid agonism can be measured by pupillometry. It is not known whether there are arteriovenous concentration differences for naloxone. The aim was thus to further develop a model for studying pharmacokinetic/pharmacodynamic aspects of naloxone and to explore whether a significant arteriovenous concentration difference for naloxone in humans was present. Relevant authorities approved this study. Healthy volunteers (n = 12) were given 1.0 mg intravenous (IV) naloxone after steady state opioid agonism was obtained by TCI of remifentanil (1.3 ng/ml). Opioid effect was measured by pupillometry. Arterial and venous samples were collected simultaneously before and for 2 h after naloxone administration for quantification of naloxone and remifentanil. Arterial remifentanil was in steady state at 12 min. One milligram IV naloxone reversed the effect of remifentanil to 93% of pre-opioid pupil-size within 4 min. The estimated duration of antagonism was 118 min. At that time, the concentration of naloxone was 0.51 ng/ml. The time course of arterial and venous serum concentrations for naloxone was similar, although arterial AUC (area under the curve) was slightly lower (94%) than the venous AUC (p = 0.03). There were no serious adverse events. Onset of reversal by IV naloxone was rapid and lasted 118 min. The minimum effective concentration was 0.5 ng/ml. Using TCI remifentanil to obtain a steady-state opioid agonism may be a useful tool to compare new naloxone products.


p27kip1 Regulates cdk2 activity in the proliferating zone of the mouse intestinal epithelium: potential role in neoplasia.
Reduced p27(kip1) expression is a marker of poor prognosis in colorectal neoplasia, and inactivation of p27 in mice (p27(Delta51/Delta51)) causes increased intestinal epithelial cell proliferation and small and large intestinal neoplasia in a diet-dependent manner. Here, we addressed the role of p27 in untransformed intestinal epithelial cells in vivo and the consequence of its targeted inactivation. A sequential fractionation procedure was used to isolate murine intestinal epithelial cells relative to their position along the crypt-villus axis, and the levels of cyclins, cyclin-dependent kinases (cdks), and cdk inhibitors and of the complexes formed among them was determined by immunoprecipitation-immunoblotting and kinase assays. As cells exited the proliferative crypt compartment, expression and activity of both cdk2 and cdk4 decreased, in parallel with reduced expression of cyclin A and proliferating cell nuclear antigen (PCNA); expression of cyclin D1, D2, and cyclin E showed little change. As expected, expression of the cdk inhibitors p21, p57, and p16 was highest in differentiated villus cells. Unexpectedly, p27 protein expression was highest in cells of the proliferative crypt compartment where it bound both cdk2 and cdk4. Cdk2 activity was increased in crypt cells from p27(Delta51/Delta51) mice, although cyclin D-associated kinase activity was unchanged (indeed, cyclin D1/2-cdk4 complex levels were reduced). Importantly, cdk2 activity was unchanged in crypt cells from p21(-/-) mice, which do not develop intestinal tumors. We propose that p27 contributes to intestinal epithelial homeostasis by regulating cdk2 activity in proliferating cells, thus gating cell cycle progression and suppressing intestinal neoplasia.


Mechanisms of phosphoryl and acyl transfer.
Acyl and phosphoryl transfer are important biochemical reactions. We have been using isotope effects caused by O-18, N-15, C-13, and deuterium substitution to examine the mechanisms and transition-state structures for enzymatic and nonenzymatic transfers of phosphoryl and acyl groups. Phosphoryl transfers from phosphate monoesters are highly dissociative, although not truly stepwise in protic solvents or in enzymatic reactions. Phosphodiesters show ANDN (SN2) reactions, whereas triester hydrolyses involve more associative transition states. Except under acidic conditions, true phosphorane intermediates likely form only when geometry requires (i.e., when the leaving group cannot be axial until pseudorotation of the phosphorane). Enzymatic phosphoryl transfers appear similar to nonenzymatic ones. The reactions of oxygen or sulfur nucleophiles with p-nitrophenyl acetate are concerted with a tetrahedral transition state, which is more dissociative with sulfur than with oxygen. Enzymatic hydrolyses of p-nitrophenyl acetate are also concerted reactions.


[Acute cardiovascular beriberi (shoshin-beriberi)].
Beriberi (BB), thiamine deficiency, has been described in the Asian literature in the 17th century and is characterized by peripheral neuropathy and muscle weakness, also called "dry" beriberi (BB) to differentiate it from "wet" BB, with essentially cardiovascular manifestations. Wet can be either "classic" wet BB in which signs and symptoms of right-sided heart failure with normal or high cardiac output are the presenting features or the "shoshin" BB variant with severe biventricular failure and metabolic acidosis, which must be treated early to prevent the rapid development of low cardiac output failure and sudden death. In this case, we report a 58 year old alcoholic woman who developed dyspnea, oliguria, edema, cardiac failure with high output, metabolic acidosis, renal tubular dysfunction and serum lactate level of 5.6 mEq/L. Neurological examination revealed peripheral neuropathy in the lower legs and cognitive alteration. She was treated with a loading dose of 100 mg of intravenous thyamine and responded with a marked increase in urine output, correction of acidosis, reduction in pulmonary-capillary wedge pressure and a change of the hemodynamic pattern. We conclude that shoshin-BB is uncommonly encountered but not widely recognized. In lactic acidosis and/or hyperdynamic circulation without any other apparent etiology in patients with possible vitamin B1 deficiency, the diagnosis of BB must be considered and thiamine should be administered.


Enlarging the amino acid set of Escherichia coli by infiltration of the valine coding pathway.
Aminoacyl transfer RNA (tRNA) synthetases establish the rules of the genetic code by catalyzing the aminoacylation of tRNAs. For some synthetases, accuracy depends critically on an editing function at a site distinct from the aminoacylation site. Mutants of Escherichia coli that incorrectly charge tRNA(Val) with cysteine were selected after random mutagenesis of the whole chromosome. All mutations obtained were located in the editing site of valyl-tRNA synthetase. More than 20% of the valine in cellular proteins from such an editing mutant organism could be replaced with the noncanonical aminobutyrate, sterically similar to cysteine. Thus, the editing function may have played a central role in restricting the genetic code to 20 amino acids. Disabling this editing function offers a powerful approach for diversifying the chemical composition of proteins and for emulating evolutionary stages of ambiguous translation.


Huanglian Jiedu Wan intervened with "Shi-Re Shanghuo" syndrome through regulating immune balance mediated by biomarker succinate.
With increasing stress in daily life and work, subhealth conditions induced by "Shi-Re Shanghuo" syndrome was gradually universal. "Huanglian Jiedu Wan" (HLJDW) was the first new syndrome Chinese medicine approved for the treatment of "Shi-Re Shanghuo" with promising clinical efficacy. Preliminary small-sample clinical studies have identified some notable biomarkers (succinate, 4-hydroxynonenal, etc.). However, the correlation and underlying mechanism between these biomarkers of HLJDW intervention on "Shi-Re Shanghuo" syndrome remained ambiguous. Therefore, this study was designed as a randomized, double-blind, multicenter, placebo-controlled Phase II clinical trial, employing integrated analysis techniques such as non-targeted and targeted metabolomics, salivary microbiota, proteomics, parallel peaction monitoring, molecular docking and surface plasmon resonance (SPR). The results of the correlation analysis indicated that HLJDW could mediate the balance between inflammation and immunity through succinate produced via host and microbial source to intervene "Shi-Re Shanghuo" syndrome. Further through the HIF1α/MMP9 pathway, succinate regulated downstream arachidonic acid metabolism, particularly the lipid peroxidation product 4-hydroxynonenal. Finally, an animal model of recurrent oral ulcers induced by "Shi-Re Shang Huo" was established and HLJDW was used for intervention, key essential indicators (succinate, glutamine, 4-hydroxynonenal, arachidonic acid metabolism) essential in the potential pathway HIF1α/MMP9 discovered in clinical practice were validated. The results were found to be consistent with our clinical findings. Taken together, succinate was observed as an important signal that triggered immune responses, which might serve as a key regulatory metabolic switch or marker of "Shi-Re Shanghuo" syndrome treated with HLJDW.


Spherical object in radiation field from a point source.
A general formula for calculating radiative fluxes from point sources of radiation incident on spherical objects was derived using some fundamental laws of classical radiometry. This formula was derived in the Cartesian coordinate system, 0xyz, where the coordinates, x, y, and z, determine the position of the spherical object with respect to the point source. The obtained solution was dependent on the radius of the object, and on the function describing the intensity of the radiation. A specific solution for calculating fluxes of isotropic radiations was presented and selected calculations were illustrated graphically.


Dimerization of the Conserved N-Terminal Domain of a Spider Silk Protein Controls the Self-Assembly of the Repetitive Core Domain.
Spider silk proteins comprise a repetitive core domain with polyalanine and glycine/proline-rich stretches flanked by highly conserved nonrepetitive N- and C-terminal domains. The termini are responsive to assembly triggers, sensing changes in the ionic (H+, phosphate) and mechanical (shear stress) environment along the spinning duct. The N-terminal domain dimerizes in a pH-dependent manner induced by protonation of conserved acidic residues. To date, dimerization of N-terminal spider silk domains has been individually investigated in the absence of large core domains. In this work, the impact of an engineered 50 kDa (AQ) core domain was studied on N-terminal dimerization by circular dichroism, fluorescence and absorbance spectroscopy, multiangle light scattering, as well as scanning electron and transmission electron microscopy. Although the core domain showed no apparent influence on the dimerization behavior of the N-terminal domain, the N-terminal domain in contrast influenced the behavior of the core domain: the monomeric state enhanced (AQ)'s solubility, and dimer formation triggered self-assembly. The monomer-dimer equilibrium was influenced by using several previously established mutants, confirming these results. This work thereby provides molecular insights into how key residues of the N-terminal domain control the dimerization-mediated transformation of soluble spidroins into fibrillary assemblies.


Development of second-generation sample enrichment probe for improved sorptive analysis of volatile organic compounds.
The sample enrichment probe (SEP) has recently been introduced as a user-friendly and cost-effective method for the sorptive extraction of volatile organic analytes from gaseous and aqueous samples for GC and GC-MS analyses. In a further development of the SEP technique, thinner polydimethylsiloxane (PDMS) tubing on polyimide-coated fused silica, instead of stainless steel rods or stalks, were used to produce the second-generation SEPs. The new SEP does not require widening of the needle-guiding orifice of the septum cap and analytes are desorbed at a faster rate from the thinner sleeve, which reduces the risk of carry-over. The flowless period that was previously recommended for analyses of highly volatile analytes is made redundant by the faster desorption from the thinner sorptive medium. It was found that differences in the thermal histories of SEPs are not the cause of the high relative standard deviations (RSDs) reported in our first paper on the technique. Excellent reproducibility can be attained by careful handling and storing of loaded SEPs and by rigorously following a standardised analytical protocol.


Development of an effective virtual environment in eliciting craving in adolescents and young adults with internet gaming disorder.
Internet gaming disorder (IGD) is a new disorder that warrants further investigation, as recently noted in the research criteria of the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition. Offering controlled environments that increase cue-induced craving, virtual reality cue-exposure therapy has been shown to be effective for some addiction disorders. To assess the feasibility of virtual reality for patients with IGD, this study aimed to develop virtual environments that represent risk situations for inducing craving, and assess the effect of virtual reality in cue reactivity. A total of 64 male adolescents and young adults (34 with IGD and 30 without) were recruited for participation. We developed a virtual internet café environment and the participants were exposed to four different tasks. As the primary feasibility outcome, cravings were measured with a visual analogue scale measuring current urge to play a game after exposure to each task. The virtual internet café induced significantly greater cravings in patients with IGD compared to controls. Additionally, patients exhibited a significantly higher acceptance rate of an avatar's invitation to play a game together than that of controls. In IGD, craving response to the tasks was positively associated with the symptom severity score as measured by Young's Internet Addiction Test. These findings reveal that virtual reality laden with complex game-related cues could evoke game craving in patients with IGD and could be used in the treatment of IGD as a cue-exposure therapy tool for eliciting craving.


Influence of plasma processing on recovery and analysis of circulating nucleic acids.
Circulating nucleic acids (CNAs) are under investigation as a liquid biopsy in cancer. However there is wide variation in blood processing and methods for isolation of circulating free DNA (cfDNA) and microRNAs (miRNAs). Here we compare the extraction efficiency and reproducibility of 4 commercially available kits for cfDNA and 3 for miRNA using spike-in of reference templates. We also compare the effects of increasing time between venepuncture and centrifugation and differential centrifugation force on recovery of CNAs. cfDNA was quantified by TaqMan qPCR and targeted deep sequencing. miRNA profiles were assessed with TaqMan low-density arrays and assays. The QIAamp(®) DNA Blood Mini and Circulating nucleic acid kits gave the highest recovery of cfDNA and efficient recovery (>90%) of a 564bp spike-in. Moreover, targeted sequencing revealed overlapping cfDNA profiles and variant depth, including detection of HER2 gene amplification, using the Ion AmpliSeq™Cancer Hotspot Panel v2. Highest yields of miRNA and the synthetic Arabidopsis thaliana miR-159a spike-in were obtained using the miRNeasy Serum/Plasma kit, with saturation above 200 µl of plasma. miRNA profiles showed significant variation with increasing time before centrifugation (p<0.001) and increasing centrifugation force, with depletion of platelet associated miRNAs, whereas cfDNA was unaffected. However, sample replicates showed excellent reproducibility on TaqMan low density arrays (ρ = 0.96, p<0.0001). We also successfully generated miRNA profiles for plasma samples stored > 12 years, highlighting the potential for analysis of stored sample biobanks. In the era of the liquid biopsy, standardisation of methods is required to minimise variation, particularly for miRNA.