Current status of liver transplantation.
Liver transplantation has become the treatment of choice for a wide range of end-stage liver disease. As outcomes have improved, so the demand for this therapy has increasingly exceeded the availability of donor organs. Access to liver transplantation is controlled such that donor organs are generally allocated to the patients who are likely to benefit most, although if all patients who might benefit were placed on the waiting list, the donor shortage would be greatly increased. Recurrence of the original liver disease is emerging as an important issue. Fewer patients are transplanted for liver tumors, as earlier results showed a very high rate of recurrence. In recent years there has been a change in the underlying conditions of patients on the waiting list, and a preponderance of patients now present with hepatitis C and alcoholic cirrhosis. Increasingly, transplant units are looking to sources of donor organs that would previously have been deemed unsuitable--such marginal donors include non-heart-beating donors (NHBDs). Results from controlled NHBDs--those cases in which cardiac arrest is predicted--suggest that this is a good source of viable organs. Splitting a donor liver to provide two grafts has successfully enabled the transplantation of a child and an adult from one organ. The transplantation of two adults from a single organ remains a greater challenge. Transplantation from living donors has been practiced increasingly over the last decade, although anxieties have been expressed over donor safety. In many countries this now represents a significant contribution to overall liver transplant activity.


TRPV4 enhances the cellular uptake of aminoglycoside antibiotics.
The cochlea and kidney are susceptible to aminoglycoside-induced toxicity. The non-selective cation channel TRPV4 is expressed in kidney distal tubule cells, and hair cells and the stria vascularis in the inner ear. To determine whether TRPV4 is involved in aminoglycoside trafficking, we generated a murine proximal-tubule cell line (KPT2) and a distal-tubule cell line (KDT3). TRPV4 expression was confirmed in KDT3 cells but not in KPT2 cells. Removal of extracellular Ca(2+) significantly enhanced gentamicin-Texas-Red (GTTR) uptake by KDT3, indicative of permeation through non-selective cation channels. To determine whether TRPV4 is permeable to GTTR, stable cell lines were generated that express TRPV4 in KPT2 (KPT2-TRPV4). KPT2-TRPV4 cells took up more GTTR than control cell lines (KPT2-pBabe) in the absence of extracellular Ca(2+). TRPV4-dependent GTTR uptake was abolished by a point mutation within the crucial pore region of the channel, suggesting that GTTR permeates the TRPV4 channel. In an endolymph-like extracellular environment, clearance of GTTR was attenuated from KPT2-TRPV4 cells in a TRPV4-dependent fashion. We propose that TRPV4 has a role in aminoglycoside uptake and retention in the cochlea.


Risk and protection factors in fatal accidents.
This paper aims at addressing the interest and appropriateness of performing accident severity analyses that are limited to fatal accident data. Two methodological issues are specifically discussed, namely the accident-size factors (the number of vehicles in the accident and their level of occupancy) and the comparability of the baseline risk. It is argued that - although these two issues are generally at play in accident severity analyses - their effects on, e.g., the estimation of survival probability, are exacerbated if the analysis is limited to fatal accident data. As a solution, it is recommended to control for these effects by (1) including accident-size indicators in the model, (2) focusing on different sub-groups of road-users while specifying the type of opponent in the model, so as to ensure that comparable baseline risks are worked with. These recommendations are applied in order to investigate risk and protection factors of car occupants involved in fatal accidents using data from a recently set up European Fatal Accident Investigation database (Reed and Morris, 2009). The results confirm that the estimated survival probability is affected by accident-size factors and by type of opponent. The car occupants' survival chances are negatively associated with their own age and that of their vehicle. The survival chances are also lower when seatbelt is not used. Front damage, as compared to other damaged car areas, appears to be associated with increased survival probability, but mostly in the case in which the accident opponent was another car. The interest of further investigating accident-size factors and opponent effects in fatal accidents is discussed.


Depression in multiple sclerosis: relationship to planning ability.
Recent research has demonstrated deficits on effortful executive tasks involving planning in multiple sclerosis (MS) patients. Given the high prevalence of depression in MS and the commonly reported link between depression and performance on executive tasks, planning impairments in MS may be associated with depression. We compared the performance of depressed and nondepressed MS patients on a planning task (Tower of London-TOL) to evaluate this hypothesis. Compared with nondepressed MS patients, depressed MS patients made significantly (p < .05) more moves and took more time per trial on the TOL. A follow-up regression analysis was conducted that included the TOL and speeded attentional/working memory task indices found to be associated with depression in MS from the authors' prior reports. This analysis revealed that 25% of the variance in depression scores was predicted by the most sensitive speeded attentional/working memory task. Furthermore, this variance overlapped completely with variance predicted by the TOL-time/trial index. The only clearly nonspeeded task index, TOL-moves per trial, was associated with unique variance (8%) in predicting MS depression scores. These results suggest that slowed information processing speed and, secondarily, deficient nonspeeded central executive skill, may be core to the cognitive deficits characteristic of depressed MS patients.


Dynamic network properties of the interictal brain determine whether seizures appear focal or generalised.
Current explanatory concepts suggest seizures emerge from ongoing dynamics of brain networks. It is unclear how brain network properties determine focal or generalised seizure onset, or how network properties can be described in a clinically-useful manner. Understanding network properties would cast light on seizure-generating mechanisms and allow to quantify to which extent a seizure is focal or generalised. Functional brain networks were estimated in segments of scalp-EEG without interictal discharges (68 people with epilepsy, 38 controls). Simplified brain dynamics were simulated using a computer model. We introduce: Critical Coupling (Cc), the ability of a network to generate seizures; Onset Index (OI), the tendency of a region to generate seizures; and Participation Index (PI), the tendency of a region to become involved in seizures. Cc was lower in both patient groups compared with controls. OI and PI were more variable in focal-onset than generalised-onset cases. In focal cases, the regions with highest OI and PI corresponded to the side of seizure onset. Properties of interictal functional networks from scalp EEG can be estimated using a computer model and used to predict seizure likelihood and onset patterns. This may offer potential to enhance diagnosis through quantification of seizure type using inter-ictal recordings.


Theoretical study of catalytic mechanism for single-site water oxidation process.
Water oxidation is a linchpin in solar fuels formation, and catalysis by single-site ruthenium complexes has generated significant interest in this area. Combining several theoretical tools, we have studied the entire catalytic cycle of water oxidation for a single-site catalyst starting with [Ru(II)(tpy)(bpm)(OH(2))](2+) (i.e., [Ru(II)-OH(2)](2+); tpy is 2,2':6',2''-terpyridine and bpm is 2,2'-bypyrimidine) as a representative example of a new class of single-site catalysts. The redox potentials and pK(a) calculations for the first two proton-coupled electron transfers (PCETs) from [Ru(II)-OH(2)](2+) to [Ru(IV) = O](2+) and the following electron-transfer process to [Ru(V) = O](3+) suggest that these processes can proceed readily in acidic or weakly basic conditions. The subsequent water splitting process involves two water molecules, [Ru(V) = O](3+) to generate [Ru(III)-OOH](2+), and H(3)O(+) with a low activation barrier (~10 kcal/mol). After the key O-O bond forming step in the single-site Ru catalysis, another PECT process oxidizes [Ru(III)-OOH](2+) to [Ru(IV)-OO](2+) when the pH is lower than 3.7. Two possible forms of [Ru(IV)-OO](2+), open and closed, can exist and interconvert with a low activation barrier (< 7 kcal/mol) due to strong spin-orbital coupling effects. In Pathway 1 at pH = 1.0, oxygen release is rate-limiting with an activation barrier ~12 kcal/mol while the electron-transfer step from [Ru(IV)-OO](2+) to [Ru(V)-OO](3+) becomes rate-determining at pH = 0 (Pathway 2) with Ce(IV) as oxidant. The results of these theoretical studies with atomistic details have revealed subtle details of reaction mechanisms at several stages during the catalytic cycle. This understanding is helpful in the design of new catalysts for water oxidation.


Fatigue is not a specific symptom in patients with primary biliary cirrhosis.
Fatigue has been reported to be common in patients with primary biliary cirrhosis (PBC). Limited data exist on comparison with fatigue in the general population and comparison with patients with other chronic gastrointestinal disorders are lacking. We enrolled 96 patients with PBC (87 females); mean age 63 (range 34-65) who completed the Fatigue Impact Scale (FIS). In comparison we included matched controls from the general population, patients with organic (OGD) and functional GI disorders (FGD). Liver function test and the latest liver biopsy were analysed and correlated with fatigue scores. The mean duration of PBC was 7.4 years, the mean bilirubin 13 micromol/l. Twelve per cent of patients had cirrhosis, 29% were in stage I on Ludwig's histology score and 30% and 29% were in stages II and III, respectively. The PBC patients had a median FIS total score of 29 in comparison with 38 in GP controls (P<0.05). Patients with OGD and FGD had more severe fatigue (FIS total score 67 and 59 (P<0.01 compared with PBC)). Fatigue in the PBC patients did not correlate with liver tests and histology stage. PBC patients had less severe fatigue measured with the FIS than controls from the GP and patients with OGD and FGD. This study also confirms results of other studies showing no correlation with fatigue in PBC and liver disease parameters. These results argues strongly against fatigue as a specific symptom in PBC.


Evaluation of human papillomavirus DNA detection in samples obtained for routine Chlamydia trachomatis screening.
The costs and logistics involved in obtaining samples is a bottleneck in large-scale studies of the circulation of human papillomavirus (HPV), which are useful for monitoring and optimisation of HPV-vaccination programs. Residual samples obtained after screening for Chlamydia trachomatis could constitute a convenient, low-cost solution. We evaluated HPV DNA detection and typing using (i) the residual samples routinely taken for C. trachomatis screening or (ii) the sample types used in large-scale phase III HPV vaccination trials (cervical, vulvar, labial, perineal, perianal, scrotal and penile shaft samples). Samples from 127 men and 110 women attending two sexual health clinics were analysed using PCR for HPV DNA, with typing using mass spectrometry. The HPV DNA prevalence was 7.1% in male urine samples, but 57.3% in female urine/vaginal samples, which was even higher than the HPV prevalence found in cervical samples (54.1%). The sensitivity for HPV DNA detection in the urine/vaginal samples was 7.9% (95% CI 3.0-16.4) for men and 78.9% (95% CI 67.6-87.7) for women, using detection in any one of the reference samples as reference. With cervical samples as reference, the sensitivity was 89.3 % (95% CI 78.1-95.9). Among men, low sensitivity of urine for HPV detection suggests limited usefulness. Among women, the high sensitivity of urine/vaginal samples for HPV detection suggests a useful low-cost solution for the study of HPV epidemiology.


Spatial segregation of different modes of movement control in the whisker representation of rat primary motor cortex.
What is mapped on the surface of the primary motor cortex (M1)? The classic somatotopic map holds true on the level of limb representations. However, on the small scale (at within-limb representations), neither somatotopy nor movement dynamics/kinematics seem to be organizational principles. We investigated the hypothesis that integrated into the body representation of M1 there may be separate representation of different modes of motor control, using different subcortical computations but sharing the same motor periphery. Using awake rats and long intracortical stimulation trains in M1 whisker representation (wM1) revealed that natural-like, rhythmic whisking (normally used for tactile exploration) can be evoked from a posteromedial subregion of wM1. Nonrhythmic whisker retraction, on the other hand, was evoked in an adjacent but more anterolaterally located region within wM1. Evoked whisker retraction was always accompanied by complex movements of the face, suggesting that the respective subregion is able to interact with other representations in specific behavioral contexts. Such associations were absent for evoked rhythmic whisking. The respective subregion rather seemed to activate a downstream central pattern generator, the oscillation frequency of which was dependent on the average evoked cortical activity. Nevertheless, joint stimulation of the two neighboring subregions demonstrated their potency to interact in a functionally useful way. Therefore, we suggest that the cause of cortical separation is the specific drive of subcortical structures needed to generate different types of movements rather than different behavioral contexts in which the movements are performed.


[Immunological aspects of narcolepsy].
Since the discovery of an almost 100 p. cent association of human narcolepsy with the HLA gene DQB1*0602, research has been focused on autoimmune mechanisms. Epidemiological data (age of onset, triggering factors, association with multiple sclerosis) would lend support to this hypothesis. However it has remained largely impossible to demonstrate immune abnormalities in blood or CSF by means of usual techniques. The canine form of the disease was supposed to be also immunologically mediated, since a linkage with a human immunoglobulin-related gene had been demonstrated. This was eventually demonstrated to be a pseudo-linkage, the real cause being a mutation in the closely related hypocretin receptor gene. This recently discovered neuropeptide is clearly involved in some aspects of sleep regulation. Soon thereafter, hypocretin deficiency was found in human narcoleptics, due to a severe neuronal loss in the hypothalamus; gliosis having been evidenced, it may be considered as the evidence of a prior inflammatory reaction, possibly due to an immune attack.


Cyanobacterial Toxins in Freshwater and Food: Important Sources of Exposure to Humans.
A recent ecological study demonstrated a significant association between an increased risk of nonalcoholic liver disease mortality and freshwater cyanobacterial blooms. Moreover, previous epidemiology studies highlighted a relationship between cyanotoxins in drinking water with liver cancer and damage and colorectal cancer. These associations identified cyanobacterial blooms as a global public health and environmental problem, affecting freshwater bodies that are important sources for drinking water, agriculture, and aquafarms. Furthermore, as a result of climate change, it is expected that our freshwater environments will become more favorable for producing harmful blooms that produce various cyanotoxins. Food is an important source of cyanotoxin exposure to humans, but it has been less addressed. This paper synthesizes information from the studies that have investigated cyanotoxins in freshwater and food on a global scale. We also review and summarize the health effects and exposure routes of cyanotoxins and candidates for cyanotoxin treatment methods that can be applied to food.


Prospective Validation of a Risk Prediction Model to Identify High-Risk Patients for Medication Errors at Hospital Admission.
Pharmacy-led medication reconciliation in elective surgery patients is often performed at the preoperative screening (POS). Because of the time lag between POS and admission, changes in medication may lead to medication errors at admission (MEAs). In a previous study, a risk prediction model for MEA was developed. To validate this risk prediction model to identify patients at risk for MEAs in a university hospital setting. The risk prediction model was derived from a cohort of a Dutch general hospital and validated within a comparable cohort from a Dutch University Medical Centre. MEAs were assessed by comparing the POS medication list with the reconciled medication list at hospital admission. This was considered the gold standard. For every patient, a risk score using the risk prediction model was calculated and compared with the gold standard. The risk prediction model was assessed with receiver operating characteristic (ROC) analysis. Of 368 included patients, 167 (45.4%) had at least 1 MEA. ROC analysis revealed significant differences in the area under the curve of 0.535 ( P = 0.26; validation cohort) versus 0.752 ( P < 0.0001; derivation cohort). The sensitivity in this validating cohort was 66%, with a specificity of 40%. Conclusion and Relevance: The risk prediction model developed in a general hospital population is not suitable to identify patients at risk for MEA in a university hospital population. However, number of medications is a common risk factor in both patient populations and should, thus, form the basis of an adapted risk prediction model.


ADAR2 Regulates Malignant Behaviour of Mesothelioma Cells Independent of RNA-editing Activity.
Malignant pleural mesothelioma (MPM) is an intractable cancer, and causes of its malignant transformation are not well known. Adenosine deaminase acting on RNA (ADAR) is an RNA-editing enzyme that converts adenosine into inosine in double-stranded RNAs potentially involved in malignant development. To examine the role of ADAR1 and ADAR2 in MPM, small interfering RNAs (siRNAs) against ADAR1 or ADAR2 were used. Transfection of siRNA against ADAR2 suppressed proliferation, motility, and invasiveness of MPM cells expressing both ADAR1 and ADAR2; however, siRNA against ADAR1 did not affect these cellular activities. Overexpression of ADAR2, that was incapable of binding to RNA, suppressed growth, motility, and invasion of MPM cells. However, overexpression of ADAR2 that had no enzyme activity did not alter the malignant properties of MPM cells. Enhancement of the malignant characteristics of cultured MPM cells via ADAR2 was independent of RNA-editing activity.


Regulation of liver glutamate dehydrogenase by reversible phosphorylation in a hibernating mammal.
Glutamate dehydrogenase (GDH) is a key enzyme that links amino acid and carbohydrate metabolism in cells. Regulation is likely most important when organisms are confronted with extreme stresses such as the low environmental temperatures and lack of food associated with winter. Many small mammals, such as Richardson's ground squirrels, Spermophilus richardsonii, cope with these conditions by hibernating. Animals enter long periods of profound torpor where metabolic rate is greatly suppressed, body temperature drops to near-ambient and all metabolic needs must be met from fixed internal body stores of fuels. To investigate how GDH is regulated under these conditions, kinetic properties of GDH were analyzed in liver from euthermic and torpid squirrels, revealing significant differences in V(max), K(m) glutamate, K(a) ADP and inhibition by urea between the two forms of GDH. These data suggested an activation of the glutamate-oxidizing activity of GDH in the hypometabolic state. Subsequent experiments suggested that the molecular basis of the kinetic differences was a change in the protein phosphorylation state of GDH between euthermia and torpor. Specifically, liver GDH appears to be dephosphorylated and activated when animals transition into torpor and this may serve to promote amino acid oxidation to contribute to energy production and gluconeogenesis. This is the first study to show that mammalian liver GDH can be regulated by reversible phosphorylation, providing an important new regulatory mechanism for GDH control.


Clinical significance of ascites in epithelial ovarian cancer.
The prognostic significance of ascites in the dissemination of metastases in epithelial ovarian cancer (EOC) is unclear. Our study aimed to investigate the association between clinicopathological factors and the development of ascites, as well as its prognostic significance. Three hundred and thirty three patients with primary EOC were suitable for inclusion. We analyzed the correlation between clinicopathological factors, including the extent of metastases, and ascitic volume. The prognostic significance of ascites was assessed using the Kaplan-Meier method and multivariate Cox's regression analysis. The average ascitic volume was 1,800 ml. Significantly, more patients with advanced FIGO stage disease presented with ascites. The volume of ascites increased significantly when metastatic disease was present in more than three regions (p<0.05), and this was the sole factor identified as associated with ascitic volume by multiple linear regression analysis. Median survival was significantly different between those with an ascitic volume less than 1,800 ml (median survival = 58 months), and those with a volume greater than 1,800 ml (median survival = 28.6 months) (p<0.05). Subgroup analysis of stage III and IV patients also revealed a poor prognosis in the presence of massive ascites (p = 0.03). Multivariate analyses found that massive ascites and poor differentiation were independent poor prognostic factors for stage III and IV EOC patients by Cox regression, using a backward elimination procedure. The volume of ascites increased significantly with the extent of metastastic disease. Massive ascites and poor tumor differentiation were associated with a worse prognosis in patients with advanced stage ovarian cancer.


The effectiveness of noninvasive interventions for musculoskeletal thoracic spine and chest wall pain: a systematic review by the Ontario Protocol for Traffic Injury Management (OPTIMa) collaboration.
The purpose of this study was to critically appraise and synthesize evidence on the effectiveness of noninvasive interventions, excluding pharmacological treatments, for musculoskeletal thoracic pain. Randomized controlled trials (RCTs), cohort studies, and case-control studies evaluating the effectiveness of noninvasive interventions were eligible. We searched MEDLINE, EMBASE, PsycINFO, and the Cochrane Central Register of Controlled Trials accessed through Ovid Technologies, Inc, and CINAHL Plus with Full Text accessed through EBSCOhost from 1990 to 2015. Our search strategies combined controlled vocabulary relevant to each database (eg, MeSH for MEDLINE) and text words relevant to our research question and the inclusion criteria. Random pairs of independent reviewers screened studies for relevance and critically appraised relevant studies using the Scottish Intercollegiate Guidelines Network criteria. Studies with a low risk of bias were synthesized following best evidence synthesis principles. We screened 6988 articles and critically appraised 2 studies. Both studies had a low risk of bias and were included in our synthesis. One RCT compared thoracic spinal manipulation, needle acupuncture, and placebo electrotherapy for recent thoracic spine pain. There were statistically significant but clinically nonimportant short-term reductions in pain favoring manipulation. There were no differences between acupuncture and placebo electrotherapy. Another RCT compared a multimodal program of care and a session of education for recent musculoskeletal chest wall pain. The multimodal care resulted in statistically significant but clinically nonimportant short-term reductions in pain over education. However, participants receiving multimodal care were more likely to report important improvements in chest pain. Quality evidence on the management of musculoskeletal thoracic pain is sparse. The current evidence suggests that compared to placebo, spinal manipulation is associated with a small and clinically nonimportant reduction in pain intensity and that acupuncture leads to similar outcomes as placebo. Furthermore, a multimodal program of care (ie, manual therapy, soft tissue therapy, exercises, heat/ice, and advice) and a single education session lead to similar pain reduction for recent-onset musculoskeletal chest wall pain. However, patients who receive multimodal care are more likely to report pain improvements.


mRNP granule proteins Fmrp and Dcp1a differentially regulate mRNP complexes to contribute to control of muscle stem cell quiescence and activation.
During skeletal muscle regeneration, satellite stem cells use distinct pathways to repair damaged myofibers or to self-renew by returning to quiescence. Cellular/mitotic quiescence employs mechanisms that promote a poised or primed state, including altered RNA turnover and translational repression. Here, we investigate the role of mRNP granule proteins Fragile X Mental Retardation Protein (Fmrp) and Decapping protein 1a (Dcp1a) in muscle stem cell quiescence and differentiation. Using isolated single muscle fibers from adult mice, we established differential enrichment of mRNP granule proteins including Fmrp and Dcp1a in muscle stem cells vs. myofibers. We investigated muscle tissue homeostasis in adult Fmr1-/- mice, analyzing myofiber cross-sectional area in vivo and satellite cell proliferation ex vivo. We explored the molecular mechanisms of Dcp1a and Fmrp function in quiescence, proliferation and differentiation in a C2C12 culture model. Here, we used polysome profiling, imaging and RNA/protein expression analysis to establish the abundance and assembly status of mRNP granule proteins in different cellular states, and the phenotype of knockdown cells. Quiescent muscle satellite cells are enriched for puncta containing the translational repressor Fmrp, but not the mRNA decay factor Dcp1a. MuSC isolated from Fmr1-/- mice exhibit defective proliferation, and mature myofibers show reduced cross-sectional area, suggesting a role for Fmrp in muscle homeostasis. Expression and organization of Fmrp and Dcp1a varies during primary MuSC activation on myofibers, with Fmrp puncta prominent in quiescence, but Dcp1a puncta appearing during activation/proliferation. This reciprocal expression of Fmrp and Dcp1a puncta is recapitulated in a C2C12 culture model of quiescence and activation: consistent with its role as a translational repressor, Fmrp is enriched in non-translating mRNP complexes abundant in quiescent myoblasts; Dcp1a puncta are lost in quiescence, suggesting stabilized and repressed transcripts. The function of each protein differs during proliferation; whereas Fmrp knockdown led to decreased proliferation and lower cyclin expression, Dcp1a knockdown led to increased cell proliferation and higher cyclin expression. However, knockdown of either Fmrp or Dcp1a led to compromised differentiation. We also observed cross-regulation of decay versus storage mRNP granules; knockdown of Fmrp enhances accumulation of Dcp1a puncta, whereas knockdown of Dcp1a leads to increased Fmrp in puncta. Taken together, our results provide evidence that the balance of mRNA turnover versus utilization is specific for distinct cellular states.


How community organizations moderate the effect of armed conflict on migration in Nepal.
This study analyses micro-level variability in migration during armed conflict in Nepal. The analysis is based on a multi-dimensional model of individual out-migration that examines the economic, social, and political consequences of conflict and how community organizations condition the experience of these consequences and systematically alter migration patterns. Detailed data on violent events and individual behaviour during the Maoist insurrection in Nepal and multi-level event-history analysis were used to test the model. The results indicate that community organizations reduced the effect of conflict on out-migration by providing resources that helped people cope with danger, as well as with the economic, social, and political consequences of the conflict. The evidence suggests that the conflict caused the population to be systematically redistributed in a way that will probably affect its future socio-demographic composition--the extent of the redistribution depending on the resources available in each community.


Historical sampling reveals dramatic demographic changes in western gorilla populations.
Today many large mammals live in small, fragmented populations, but it is often unclear whether this subdivision is the result of long-term or recent events. Demographic modeling using genetic data can estimate changes in long-term population sizes while temporal sampling provides a way to compare genetic variation present today with that sampled in the past. In order to better understand the dynamics associated with the divergences of great ape populations, these analytical approaches were applied to western gorillas (Gorilla gorilla) and in particular to the isolated and Critically Endangered Cross River gorilla subspecies (G. g. diehli). We used microsatellite genotypes from museum specimens and contemporary samples of Cross River gorillas to infer both the long-term and recent population history. We find that Cross River gorillas diverged from the ancestral western gorilla population ~17,800 years ago (95% HDI: 760, 63,245 years). However, gene flow ceased only ~420 years ago (95% HDI: 200, 16,256 years), followed by a bottleneck beginning ~320 years ago (95% HDI: 200, 2,825 years) that caused a 60-fold decrease in the effective population size of Cross River gorillas. Direct comparison of heterozygosity estimates from museum and contemporary samples suggests a loss of genetic variation over the last 100 years. The composite history of western gorillas could plausibly be explained by climatic oscillations inducing environmental changes in western equatorial Africa that would have allowed gorilla populations to expand over time but ultimately isolate the Cross River gorillas, which thereafter exhibited a dramatic population size reduction. The recent decrease in the Cross River population is accordingly most likely attributable to increasing anthropogenic pressure over the last several hundred years. Isolation of diverging populations with prolonged concomitant gene flow, but not secondary admixture, appears to be a typical characteristic of the population histories of African great apes, including gorillas, chimpanzees and bonobos.


Hormones, menstrual distress, and migraine across the phases of the menstrual cycle.
The primary objectives of the present study were to (1) contrast reproductive hormone levels and ratings of menstrual distress of female migraineurs with those of a control group in each menstrual cycle phase, (2) examine correlations between hormone levels and migraine frequency, severity, and migraine-related disability, and (3) examine correlations between menstrual distress and migraine frequency, severity, and migraine-related disability. A secondary objective was to evaluate the validity of a migraine disability measure modified to reflect 7-day recall. Further controlled, prospective study is needed regarding the temporal relationships between reproductive hormones at each stage of the menstrual cycle and fluctuations in migraine activity across the cycle. Twenty-three women (17 with migraine, 6 control participants) completed laboratory hormone assays and measures of menstrual distress and disability at each phase of one menstrual cycle, and monitored their headache activity daily during the same cycle. Results.-The migraine group evidenced lower premenstrual luteinizing hormone and more menstrual distress symptoms at each phase of the menstrual cycle. Hormones were associated with migraine activity and disability within cycle phases, and across phases in a time-lagged manner. Menstrual distress was associated with ovulatory phase migraine activity and with migraine-related disability across the menstrual cycle. A retrospective 7-day migraine disability measure appeared to be a consistently valid index. Both reproductive hormones and menstrually related distress appear to predict migraine activity and disability. These associations were evident not only for perimenstrual migraine, but also for migraine at each phase of the menstrual cycle.


Mental health of indigenous school children in Northern Chile.
Anxiety and depressive disorders occur in all stages of life and are the most common childhood disorders. However, only recently has attention been paid to mental health problems in indigenous children and studies of anxiety and depressive disorders in these children are still scarce. This study compares the prevalence of anxiety and depressive symptoms in Aymara and non-Aymara children. Among the Aymara children, the study examines the relations between these symptoms and the degree of involvement with Aymara culture. We recruited 748 children aged 9 to 15 years from nine schools serving low socioeconomic classes in the city of Arica, in northern Chile. The children were equally divided between boys and girls and 37% of the children were Aymara. To evaluate anxiety and depressive symptoms we used the Stress in Children (SiC) instrument and the Children Depression Inventory-Short version (CDI-S), and used an instrument we developed to assess level of involvement in the Aymara culture. There was no significant difference between Aymara and non-Aymara children on any of the instrument scales. Dividing the Aymara children into high-involvement (n = 89) and low-involvement (n = 186) groups, the low-involvement group had significantly higher scores on the Hopelessness subscale of the CDI-S (p = 0.02) and scores of marginally higher significance in overall Anxiety on the SiC (p = 0.06). Although Aymara children have migrated from the high Andean plateau to the city, this migration has not resulted in a greater presence of anxiety and depressive symptoms. Greater involvement with the Aymara culture may be a protective factor against anxiety and depressive symptoms in Aymara children. This point to an additional benefit of maintaining cultural traditions within this population.


Learning-based vertebra detection and iterative normalized-cut segmentation for spinal MRI.
Automatic extraction of vertebra regions from a spinal magnetic resonance (MR) image is normally required as the first step to an intelligent spinal MR image diagnosis system. In this work, we develop a fully automatic vertebra detection and segmentation system, which consists of three stages; namely, AdaBoost-based vertebra detection, detection refinement via robust curve fitting, and vertebra segmentation by an iterative normalized cut algorithm. In order to produce an efficient and effective vertebra detector, a statistical learning approach based on an improved AdaBoost algorithm is proposed. A robust estimation procedure is applied on the detected vertebra locations to fit a spine curve, thus refining the above vertebra detection results. This refinement process involves removing the false detections and recovering the miss-detected vertebrae. Finally, an iterative normalized-cut segmentation algorithm is proposed to segment the precise vertebra regions from the detected vertebra locations. In our implementation, the proposed AdaBoost-based detector is trained from 22 spinal MR volume images. The experimental results show that the proposed vertebra detection and segmentation system can achieve nearly 98% vertebra detection rate and 96% segmentation accuracy on a variety of testing spinal MR images. Our experiments also show the vertebra detection and segmentation accuracies by using the proposed algorithm are superior to those of the previous representative methods. The proposed vertebra detection and segmentation system is proved to be robust and accurate so that it can be used for advanced research and application on spinal MR images.


Online treatment of persistent complex bereavement disorder, posttraumatic stress disorder, and depression symptoms in people who lost loved ones during the COVID-19 pandemic: study protocol for a randomized controlled trial and a controlled trial.
Losing a loved one during the COVID-19 pandemic is a potentially traumatic loss that may result in symptoms of persistent complex bereavement disorder (PCBD), posttraumatic stress disorder (PTSD), and depression. To date, grief-specific cognitive-behavioural therapy (CBT) has mostly been delivered through individual face-to-face formats, while studies have shown that online treatment also yields promising results. Offering treatment online is now more than ever relevant during the pan demic and may offer important benefits compared with face-to-face CBT, such as lower costs and higher accessibility. Our expectation is that grief-specific online CBT is effective in reducing PCBD, PTSD, and depression symptoms. Our aim is to evaluate the short-term and long-term effectiveness of grief-specific online CBT in reducing PCBD, PTSD, and depression symptom-levels for adults who lost a loved one during the COVID-19 pandemic. This study consists of two parts. In part 1, a two-armed (unguided online CBT versus waitlist controls) randomized controlled trial will be conducted. In part 2, a two-armed (guided online CBT versus unguided online CBT) controlled trial will be conducted. Symptoms of PCBD, PTSD, and depression will be assessed via telephone interviews at pre-treatment/pre-waiting period, post-treatment/post-waiting period, and six months post-treatment. Potential participants are people who lost a loved one at least three months earlier during the COVID-19 pandemic with clinically relevant levels of PCBD, PTSD, and/or depression. Analysis of covariance and multilevel modelling will be performed. This is one of the first studies examining the effectiveness of online grief-specific CBT. More research is needed before implementing online grief-specific CBT into clinical practice.


Optovibrometry: tracking changes in the surface tension and viscosity of multicomponent droplets in real-time.
An instrument was developed for measuring real time changes in the surface tension and viscosity of multicomponent droplets of miscible liquids and other soft materials. Droplets containing glycerol and water were supported on superamphiphobic surfaces and vibrated by applying a short mechanical impulse. Laser light was refracted through the droplets and allowed to fall on the surface of a photodiode. Time dependent variations in the intensity measured by the photodiode during vibration were used to monitor the decay of the droplet oscillations. The frequencies and spectral widths of the droplet vibrational resonances were then obtained from Fourier transforms of these time dependent intensity signals. A recently developed model of viscoelastic droplet vibration was used along with these values and measurements of the drop dimensions to extract the surface tension and viscosity of the drops as they evaporated. Collection of data was automated and values of frequency, spectral width, drop size, surface tension and viscosity were obtained with a time resolution of three seconds over a period of thirty minutes. The values of surface tension and viscosity obtained were shown to be in good agreement with literature values obtained from bulk glycerol/water solutions; thus validating the technique for wider application to other multicomponent liquids and soft matter systems.


Presence of pleural effusion is associated with a poor prognosis in patients with epidermal growth factor receptor-mutated lung cancer receiving tyrosine kinase inhibitors as first-line treatment.
This study was conducted to evaluate the effect of clinical factors on the treatment outcomes of lung cancer patients with active epidermal growth factor receptor (EGFR) mutations treated by first-line tyrosine kinase inhibitors (TKIs). Patients of stage IIIb or IV lung adenocarcinoma harboring mutated EGFR were enrolled between March 2010 and June 2014 and followed up until December 2015. The effects of various clinical features, such as age, sex, smoking history, EGFR mutation types, TKIs used, presence of pleural effusion, metastatic sites on progression-free survival (PFS) and overall survival (OS), were analyzed retrospectively. A total of 104 patients were included in this study. Patients with pleural effusion at initial diagnosis had significantly shorter PFS and OS than those without pleural effusion (median PFS: 8.2 months vs 15.3 months, P = 0.0004; median OS: 16.3 months vs 28.2 months, P = 0.0003). Univariate analysis revealed that being male or a smoker was associated with short PFS, whereas smoking history, bony metastasis and malignant pleural effusion were associated with poor OS. Stepwise multivariate Cox regression analysis showed that the presence of pleural effusion and different TKI use were independent prognostic factors for PFS [hazard ratio [HR] = 2.50 (95% confidence interval [CI], 1.53-4.10), P = 0.0003 and HR = 0.55 (95% CI, 0.31-0.97), P = 0.0396, respectively], whereas the presence of pleural effusion and liver metastasis were associated with poor OS [HR = 2.79 (95% CI: 1.46-5.30), P = 0.0018 and HR = 2.12 (95% CI, 1.02-4.40), P = 0.0440, respectively]. The presence of pleural effusion predicts poor PFS and OS in lung adenocarcinoma patients receiving TKIs as the first-line treatment. Additional studies are warranted to elucidate the underlying mechanisms and determine novel strategies for improving the outcome of these patients.


Comparison of thyromental height test with ratio of height to thyromental distance, thyromental distance, and modified Mallampati test in predicting difficult laryngoscopy: A prospective study.
Preoperative airway assessment to predict patients with difficult laryngoscopy is always crucial for anesthesiologists. Several predictive tests have been studied by various authors in quest of finding the best airway predictor. Recently, a new airway predictor, thyromental height test (TMHT) has been reported to have good predictive value in assessing difficult airway. We conducted this study with primary aim to evaluate the diagnostic accuracy of TMHT and to compare it with other established airway predictors, such as ratio of height to thyromental distance (RHTMD), thyromental distance (TMD), and modified Mallampati test (MMT) for predicting difficult laryngoscopy. This prospective, observational study was conducted in 550 patients of either sex aged >18 years scheduled for elective surgery under general anesthesia. The patients' airway was assessed preoperatively by two anesthetists. Standard anesthetic protocol was followed in all the patients. The laryngoscopic view was graded according to Cormack-Lehane scale. The receiver operating characteristic (ROC) curve was used to calculate the ideal cut off values for TMHT and RHTMD. Standard formulae were used to calculate validity indexes. The incidence of difficult laryngoscopy was 10%. The cut-off value for TMHT and RHTMD were 5.1 cm and 19.5, respectively. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of TMHT were 78.18%, 93.94%, 58.90%, and 97.48%, respectively. The highest sensitivity, PPV, and NPV were observed with TMHT as compared with RHTMD, TMD, and MMT (P < 0.0001). TMHT is the best predictive test with highest accuracy and odds ratio for predicting difficult airway out of all predictive tests evaluated.


Harnessing the wisdom of crowds can improve guideline compliance of antibiotic prescribers and support antimicrobial stewardship.
Antibiotic overprescribing is a global challenge contributing to rising levels of antibiotic resistance and mortality. We test a novel approach to antibiotic stewardship. Capitalising on the concept of "wisdom of crowds", which states that a group's collective judgement often outperforms the average individual, we test whether pooling treatment durations recommended by different prescribers can improve antibiotic prescribing. Using international survey data from 787 expert antibiotic prescribers, we run computer simulations to test the performance of the wisdom of crowds by comparing three data aggregation rules across different clinical cases and group sizes. We also identify patterns of prescribing bias in recommendations about antibiotic treatment durations to quantify current levels of overprescribing. Our results suggest that pooling the treatment recommendations (using the median) could improve guideline compliance in groups of three or more prescribers. Implications for antibiotic stewardship and the general improvement of medical decision making are discussed. Clinical applicability is likely to be greatest in the context of hospital ward rounds and larger, multidisciplinary team meetings, where complex patient cases are discussed and existing guidelines provide limited guidance.


Comparative analysis of the infectivity rate of both Borrelia burgdorferi and Anaplasma phagocytophilum in humans and dogs in a New Jersey community.
Ticks are important vectors of disease and transmit an extensive array of bacterial, viral and protozoan diseases to both humans and dogs within a community. Borrelia burgdorferi, the causative agent of Lyme disease, has been extensively studied within both the human and veterinary population. Anaplasma phagocytophilum, an intracellular rickettsial pathogen also transmitted by ixodid ticks, has emerged as an important zoonotic infection with significant veterinary and medical implications, and is responsible for both canine granulocytic anaplasmosis and human granulocytic anaplasmosis. Multiple surveys exist in the international literature referencing infectivity rates of both of these diseases separately in both the dog and human populations. This is the first study to simultaneously examine the infectivity rate of both anaplasmosis and Lyme disease in humans and dogs in a community endemic for tick-borne diseases.


Effect of epimestrol on gonadotropin and prolactin plasma levels and response to luteinizing hormone-releasing hormone/thyrotropin-releasing hormone in secondary amenorrhea and oligomenorrhea.
The effects of epimestrol (5 mg every 6 hours for 5 days) on basal levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin (Prl), estradiol, progesterone, and dehydroepiandrosterone sulfate, and on the response to LH-releasing hormone (LH-RH) and thyrotropin-releasing hormone (TRH) stimulation, were studied in 18 cases of secondary amenorrhea and oligomenorrhea of hypothalamic-pituitary origin, in three cases of anorexia nervosa, in two cases of long-lasting progestin-induced amenorrhea, and in one case of precocious menopause. The results in the first 18 patients indicate that epimestrol treatment induces a significant increase in LH and Prl levels after 24 hours, while the FSH increase becomes significant only after 4 days of therapy. Twelve hours after discontinuation of treatment, all three hormone levels decreased significantly to values similar to the basal levels, while the pituitary response to LH-RH indicated a much more marked LH secretion than before treatment. A second test, performed 36 hours after the last drug administration, again showed a significantly higher LH response than that found under basal conditions. No significant variations were observed in the FSH response to LH-RH, nor in the Prl response to TRH. These data suggest that epimestrol interferes at the level of the centers responsible for Prl and gonadotropin secretion in the manner of a weak estrogen.


Effects of HIV-1 infection on malaria parasitemia in milo sub-location, western Kenya.
Malaria and HIV infections are both highly prevalent in sub-Saharan Africa, with HIV-infected patients being at higher risk of acquiring malaria. HIV-1 infection is known to impair the immune response and may increase the incidence of clinical malaria. However, a positive association between HIV-1 and malaria parasitaemia is still evolving. Equally, the effect of malaria on HIV-1 disease stage has not been well established, but when fever and parasitemia are high, malaria may be associated with transient increases in HIV-1 viral load, and progression of HIV-1 asymptomatic disease phase to AIDS. To determine the effects of HIV-1 infection on malaria parasitaemia among consented residents of Milo sub-location, Bungoma County in western Kenya. Census study evaluating malaria parasitaemia in asymptomatic individuals with unknown HIV-1 status. After ethical approvals from both Moi University and MTRH research ethics committees, data of 3,258 participants were retrieved from both Webuye health demographic surveillance system (WHDSS), and Academic Model Providing Access to Healthcare (AMPATH) in the year 2010. The current study was identifying only un-diagnosed HIV-1 individuals at the time the primary data was collected. The data was then analysed for significant statistical association for malaria parasitemia and HIV-1 infection, using SPSS version 19. Demographic characteristics such as age and sex were summarized as means and percentages, while relationship between malaria parasitaemia and HIV-1 (serostatus) was analyzed using Chi square. Age distribution for the 3,258 individuals ranged between 2 and 94 years, with a mean age of 26 years old. Females constituted 54.3%, while males were 45.8%. In terms of age distribution, 2-4 years old formed 15.1% of the study population, 5-9 years old were 8.8%, 10-14 years old were 8.6% while 15 years old and above were 67.5%. Of the 3,258 individuals whose data was eligible for analysis, 1.4% was newly diagnosed HIV-1 positive. Our findings showed a higher prevalence of malaria in children aged 2-10 years (73.4%), against the one reported in children in lake Victoria endemic region by the Kenya malaria indicator survey in the year 2010 (38.1%). There was no significant associations between the prevalence of asymptomatic malaria and HIV-1 status (p = 0.327). However, HIV-1/malaria co-infected individuals showed elevated mean malaria parasite density, compared to HIV-1 negative individuals, p = 0.002. HIV-1 status was not found to have effect on malaria infection, but the mean malaria parasite density was significantly higher in HIV-1 positive than the HIV-1 negative population.


The accumulation of cosmogenic chlorine-36 in rocks: a method for surface exposure dating.
Because of the reaction of cosmic rays with elements in minerals, chlorine-36 accumulates in rocks exposed at the earth's surface. This suggests that the ratio of chlorine-36 to stable chloride can be used as a geochronometer. Reasonable agreement has been obtained between measurements of chlorine-36 in volcanic rocks of known age and calculations of chlorine-36 production based on rock composition and cosmicray flux. The buildup of chlorine-36 should be a useful means for measuring the surface exposure time of young volcanic rocks and geomorphic features in the age range of 10(3) to 10(6) years.


The Dutch public are positive about the colorectal cancer-screening programme, but is this a well-informed opinion?
Population-based colorectal cancer (CRC) screening is widely recommended, and members of the eligible screening population seem to be positive about it. However, it is not well known how people outside the eligible screening population view CRC screening, and whether they are supportive of the government providing this. Public opinion may affect people's personal views and their screening decision. The aim of our study was to examine the opinion of the Dutch general public regarding the national CRC screening programme. An online survey was carried out in a Dutch population sample of adults aged 18 and older, assessing level of support, personal attitude, collective attitude, perceived social norm, awareness, and knowledge regarding the CRC screening programme. The response rate was 56% (n = 1679/3000). Generally, the Dutch public are positive about and supportive of the CRC screening programme. We found the biggest proportion of support (86%) when people were asked directly. A smaller proportion (48%) was supportive when people had to choose between other options concerning how the government could possibly deal with CRC. People report knowing more about the benefits of CRC screening than about its possible harms and risks. Many people found it difficult to answer the knowledge questions that asked about numerical information concerning CRC screening correctly. People were less supportive of the CRC screening programme when having to choose between other options concerning dealing with CRC, and their support may not be based on a full comprehension of what CRC screening entails. Further research is needed to establish what knowledge people need in order to form a well-founded opinion.


Large-Scale Screening of 239 Traditional Chinese Medicinal Plant Extracts for Their Antibacterial Activities against Multidrug-Resistant Staphylococcus aureus and Cytotoxic Activities.
Novel alternative antibacterial compounds have been persistently explored from plants as natural sources to overcome antibiotic resistance leading to serious foodborne bacterial illnesses. In this study, the ethanolic extracts from 239 traditional Chinese medicinal plants (TCMP)' materials were screened to discover promising candidates that have strong antibacterial properties against multidrug-resistant Staphylococcus (S.) aureus and low cytotoxicity. The results revealed that 74 extracts exhibited good antibacterial activities (diameter of inhibition zone (DIZ) ≥ 15 mm). Furthermore, 18 extracts (DIZ ≥ 20 mm) were determined their minimum inhibitory concentrations (MIC) and minimum bactericide concentrations (MBC), ranging from 0.1 to 12.5 mg/mL and 0.78 to 25 mg/mL, respectively. In addition, most of the 18 extracts showed relatively low cytotoxicity (a median lethal concentration (LC50) >100 µg/mL). The 18 extracts were further determined to estimate possible correlation of their phenolic contents with antibacterial activity, and the results did not show any significant correlation. In conclusion, this study selected out some promising antibacterial TCMP extracts with low cytotoxicity, including Rhus chinensis Mill., Ilex rotunda Thunb., Leontice kiangnanensis P.L.Chiu, Oroxylum indicum Vent., Isatis tinctorial L., Terminalia chebula Retz., Acacia catechu (L.f.) Willd., Spatholobus suberectus Dunn, Rabdosia rubescens (Hemsl.) H.Hara, Salvia miltiorrhiza Bunge, Fraxinus fallax Lingelsh, Coptis chinensis Franch., Agrimonia Pilosa Ledeb., and Phellodendron chinense C.K.Schneid.


Assembly-disassembly is coupled to the ATPase cycle of tobacco Rubisco activase.
The carbon-fixing activity of enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is regulated by Rubisco activase (Rca), a ring-forming ATPase that catalyzes inhibitor release. For higher plant Rca, the catalytic roles played by different oligomeric species have remained obscure. Here, we utilized fluorescence-correlation spectroscopy to estimate dissociation constants for the dimer-tetramer, tetramer-hexamer, hexamer-12-mer, and higher-order assembly equilibria of tobacco Rca. A comparison of oligomer composition with ATPase activity provided evidence that assemblies larger than hexamers are hydrolytically inactive. Therefore, supramolecular aggregates may serve as storage forms at low-energy charge. We observed that the tetramer accumulates only when both substrate and product nucleotides are bound. During rapid ATP turnover, about one in six active sites was occupied by ADP, and ∼36% of Rca was tetrameric. The steady-state catalytic rate reached a maximum between 0.5 and 2.5 μm Rca. In this range, significant amounts of dimers, tetramers, and hexamers coexisted, although none could fully account for the observed activity profile. Therefore, we propose that dynamic assembly-disassembly partakes in the ATPase cycle. According to this model, the association of dimers with tetramers generates a hexamer that forms a closed ring at high ATP and magnesium levels. Upon hydrolysis and product release, the toroid breaks open and dissociates into a dimer and tetramer, which may be coupled to Rubisco remodeling. Although a variant bearing the R294V substitution assembled in much the same way, highly stabilized states could be generated by binding of a transition-state analog. A tight-binding pre-hydrolysis state appears to become more accessible in thermally labile Rcas.


PDL1 expression is an independent prognostic factor in localized GIST.
Gastrointestinal stromal tumors (GIST) are the most frequently occurring digestive sarcomas. The prognosis of localized GIST is heterogeneous, notably for patients with an Armed Forces Institute of Pathology (AFIP) intermediate or high risk of relapse. Despite imatinib effectiveness, it is crucial to develop therapies able to overcome the resistance mechanisms. The immune system represents an attractive prognostic and therapeutic target. The Programmed cell Death 1 (PD1)/programmed cell death ligand 1 (PDL1) pathway is a key inhibitor of the immune response; recently, anti-PD1 and anti-PDL1 drugs showed very promising results in patients with solid tumors. However, PDL1 expression has never been studied in GIST. Our objective was to analyze PDL1 expression in a large series of clinical samples. We analyzed mRNA expression data of 139 operated imatinib-untreated localized GIST profiled using DNA microarrays and searched for correlations with histoclinical features including postoperative metastatic relapse. PDL1 expression was heterogeneous across tumors and was higher in AFIP low-risk than in high-risk samples, and in samples without than with metastatic relapse. PDL1 expression was associated with immunity-related parameters such as T-cell-specific and CD8+ T-cell-specific gene expression signatures and probabilities of activation of interferon α (IFNα), IFNγ, and tumor necrosis factor α (TNFα) pathways, suggesting positive correlation with a cytotoxic T-cell response. In multivariate analysis, the PDL1-low group was associated with a higher metastatic risk independently of the AFIP classification and the KIT mutational status. In conclusion, PDL1 expression refines the prediction of metastatic relapse in localized GIST and might improve our ability to better tailor adjuvant imatinib. In the metastatic setting, PDL1 expression might guide the use of PDL1 inhibitors, alone or associated with tyrosine kinase inhibitors.


Quantitative metabolomics by H-NMR and LC-MS/MS confirms altered metabolic pathways in diabetes.
Insulin is as a major postprandial hormone with profound effects on carbohydrate, fat, and protein metabolism. In the absence of exogenous insulin, patients with type 1 diabetes exhibit a variety of metabolic abnormalities including hyperglycemia, glycosurea, accelerated ketogenesis, and muscle wasting due to increased proteolysis. We analyzed plasma from type 1 diabetic (T1D) humans during insulin treatment (I+) and acute insulin deprivation (I-) and non-diabetic participants (ND) by (1)H nuclear magnetic resonance spectroscopy and liquid chromatography-tandem mass spectrometry. The aim was to determine if this combination of analytical methods could provide information on metabolic pathways known to be altered by insulin deficiency. Multivariate statistics differentiated proton spectra from I- and I+ based on several derived plasma metabolites that were elevated during insulin deprivation (lactate, acetate, allantoin, ketones). Mass spectrometry revealed significant perturbations in levels of plasma amino acids and amino acid metabolites during insulin deprivation. Further analysis of metabolite levels measured by the two analytical techniques indicates several known metabolic pathways that are perturbed in T1D (I-) (protein synthesis and breakdown, gluconeogenesis, ketogenesis, amino acid oxidation, mitochondrial bioenergetics, and oxidative stress). This work demonstrates the promise of combining multiple analytical methods with advanced statistical methods in quantitative metabolomics research, which we have applied to the clinical situation of acute insulin deprivation in T1D to reflect the numerous metabolic pathways known to be affected by insulin deficiency.


DNA Replication Determines Timing of Mitosis by Restricting CDK1 and PLK1 Activation.
To maintain genome stability, cells need to replicate their DNA before dividing. Upon completion of bulk DNA synthesis, the mitotic kinases CDK1 and PLK1 become active and drive entry into mitosis. Here, we have tested the hypothesis that DNA replication determines the timing of mitotic kinase activation. Using an optimized double-degron system, together with kinase inhibitors to enforce tight inhibition of key proteins, we find that human cells unable to initiate DNA replication prematurely enter mitosis. Preventing DNA replication licensing and/or firing causes prompt activation of CDK1 and PLK1 in S phase. In the presence of DNA replication, inhibition of CHK1 and p38 leads to premature activation of mitotic kinases, which induces severe replication stress. Our results demonstrate that, rather than merely a cell cycle output, DNA replication is an integral signaling component that restricts activation of mitotic kinases. DNA replication thus functions as a brake that determines cell cycle duration.


Comparison Between the SFTS-QS Kit and the PowerChek SFTSV Real-time PCR Kit for the Detection of Severe Fever With Thrombocytopenia Syndrome Virus.
The recent increase in severe fever with thrombocytopenia syndrome (SFTS) cases has led to the development of the SFTS-QS kit (MiCoBioMed, Seongnam, Korea) for detecting the SFTS virus (SFTSV, now renamed Huaiyangshan banyangvirus). SFTS-QS is a qualitative real-time reverse transcription PCR assay based on lab-on-a-chip technology. We evaluated the performance of the SFTS-QS kit and compared it with that of the PowerChek SFTSV Real-time PCR kit (PowerChek; Kogene Biotech, Seoul, Korea). A total of 117 serum samples were simultaneously assayed using the SFTS-QS and PowerChek kits. Sanger sequencing targeting the S and M segments of SFTSV was performed as the reference method. The total turnaround time of the two kits was compared. The SFTS-QS results agreed with those of PowerChek with a kappa value of 0.92. The diagnostic sensitivity and specificity of the SFTS-QS kit were both 100% (14/14 and 103/103, respectively), whereas those of the PowerChek kit were 100% (14/14) and 98.1% (101/103), respectively. The results of SFTS-QS and PowerChek were comparable; however, the SFTS-QS kit required a shorter total turnaround time. The SFTS-QS kit produced accurate and fast results and thus could serve as a useful tool for detecting SFTSV.


A robust tool highlights the influence of bird migration on influenza A virus evolution.
One of the fundamental unknowns in the field of influenza biology is a panoramic understanding of the role wild birds play in the global maintenance and spread of influenza A viruses. Wild aquatic birds are considered a reservoir host for all lowly pathogenic avian influenza A viruses (AIV) and thus serve as a potential source of zoonotic AIV, such as Australasian-origin H5N1 responsible for morbidity and mortality in both poultry and humans, as well as genes that may contribute to the emergence of pandemic viruses. Years of broad, in-depth wild bird AIV surveillance have helped to decipher key observations and ideas regarding AIV evolution and viral ecology including the trending of viral lineages, patterns of gene flow within and between migratory flyways and the role of geographic boundaries in shaping viral evolution (Bahl et al. 2009; Lam et al. 2012). While these generally 'virus-centric' studies have ultimately advanced our broader understanding of AIV dynamics, recent studies have been more host-focused, directed at determining the potential impact of host behaviour on AIV, specifically, the influence of bird migration upon AIV maintenance and transmission. A large number of surveillance studies have taken place in Alaska, United States-a region where several global flyways overlap-with the aim of detecting the introduction of novel, Australasian-origin highly pathogenic H5N1 AIV into North America. By targeting bird species with known migration habits, long-distance migrators were determined to be involved in the intercontinental movement of individual AIV gene segments, but not entire viruses, between the Australasian and North American flyways (Koehler et al. 2008; Pearce et al. 2010). Yet, bird movement is not solely limited to long-distance migration, and the relationship of resident or nonmigratory and intermediate-distance migrant populations with AIV ecology has only recently been explored by Hill et al. (2012) in this issue of Molecular Ecology. Applying a uniquely refined, multidimensional approach, Hill et al. validate the innovative use of stable isotope assays for qualifying migration status of wild mallards within the Pacific flyway. The authors reveal that AIV prevalence and diversity did not differ in wintering mallard ducks with different migration strategies, and while migrant mallards do indeed introduce AIV, these viruses do not circulate as the predominant viruses in resident birds. On the other hand, resident mallards from more temperate regions act as reservoirs, possibly contributing to the unseasonal circulation and extended transmission period of AIV. This study highlights the impact of animal behaviour on shaping viral evolution, and the unique observations made will help inform prospective AIV surveillance efforts in wild birds.


The monocyte differentiation antigen, CD14, is anchored to the cell membrane by a phosphatidylinositol linkage.
CD14 is a myeloid differentiation Ag expressed primarily on peripheral blood monocytes and macrophages. Although its function is unknown, the CD14 gene maps to a region encoding several myeloid growth factors and receptors. Analysis of the CD14 protein sequence deduced from the cDNA shows that although the CD14 protein contains a characteristic leader peptide, it lacks a characteristic transmembrane region, suggesting that CD14 may be anchored to the membrane via glycosylphosphatidylinositol (PI). Treatment of monocytes as well as a CD14-expressing neuroglioma cell line with PI-phospholipase C removed CD14 from the cell surface. Furthermore, monocytes from a patient with paroxysmal nocturnal hemoglobinuria, a disease characterized by lack of expression of other PI-linked proteins, failed to express CD14. Interestingly, the CD14-expressing neuroglioma cell line, which had been transfected with a single CD14 cDNA, released a soluble form of CD14 into the supernatant. Soluble forms of CD14 have previously been observed in serum of normal individuals and in culture supernatants of CD14+ cells. Biosynthetic experiments reveal that this soluble form of CD14 (48 kDa), which is smaller than the form released from the membrane by PI-phospholipase C (53 kDa), does not contain ethanolamine, the first constitutent of the PI-anchoring system. These studies demonstrate that CD14 is a member of the family of PI-anchored proteins and suggest that soluble forms of CD14 represent molecules that completely lack the PI-anchoring system.


Daily Enteral DHA Supplementation Alleviates Deficiency in Premature Infants.
Docosahexaenoic acid (DHA) is an essential fatty acid (FA) important for health and neurodevelopment. Premature infants are at risk of DHA deficiency and circulating levels directly correlate with health outcomes. Most supplementation strategies have focused on increasing DHA content in mother's milk or infant formula. However, extremely premature infants may not reach full feedings for weeks and commercially available parenteral lipid emulsions do not contain preformed DHA, so blood levels decline rapidly after birth. Our objective was to develop a DHA supplementation strategy to overcome these barriers. This double-blind, randomized, controlled trial determined feasibility, tolerability and efficacy of daily enteral DHA supplementation (50 mg/day) in addition to standard nutrition for preterm infants (24-34 weeks gestational age) beginning in the first week of life. Blood FA levels were analyzed at baseline, full feedings and near discharge in DHA (n = 31) or placebo supplemented (n = 29) preterm infants. Term peers (n = 30) were analyzed for comparison. Preterm infants had lower baseline DHA levels (p < 0.0001). Those receiving DHA had a progressive increase in circulating DHA over time (from 3.33 to 4.09 wt% or 2.88 to 3.55 mol%, p < 0.0001) while placebo-supplemented infants (receiving standard neonatal nutrition) had no increase over time (from 3.35 to 3.32 wt% or 2.91 to 2.87 mol%). Although levels increased with additional DHA supplementation, preterm infants still had lower blood DHA levels than term peers (4.97 wt% or 4.31 mol%) at discharge (p = 0.0002). No differences in adverse events were observed between the groups. Overall, daily enteral DHA supplementation is feasible and alleviates deficiency in premature infants.


Single-wavelength-controlled in situ dynamic super-resolution fluorescence imaging for block copolymer nanostructures via blue-light-switchable FRAP.
Photoswitchable fluorophores are promising in single-molecule optical devices and super-resolution fluorescence imaging, especially in single-molecule photo-activated localization microscopy (PALM) or stochastic optical reconstruction microscopy (STORM). However, the scarcity of current photoswitchable fluorophores stimulates researchers to develop complicated optical systems and processing software, in accordance with the limited photoswitchable fluorescent proteins and organic fluorophores. Previous efforts to develop synthetic photoswitchable fluorophores have exhibited their promising potential in super-resolution fluorescence imaging. Here, we have designed and synthesized a fluorescence molecular switch with reversible green emission, a napthalimide-hexaarylbiimidazole conjugate (NI-N-HABI), which exhibits strong fluorescence in the emissive state, with fast thermal fading of the photochromism and spontaneous fluorescence recovery after photobleaching (FRAP) induced by blue-light. The photoswitchable fluorophore enables the red-edge wavelength of the optical response to red-shift from the initial near-UV region at less than 400 nm, to 500 nm. The relatively fast fading speed of NI-N-HABI and its sensitivity to longer blue-light irradiation (400-500 nm) have allowed simplification of the optical microscopic system from a two-wavelength laser source to a single-wavelength laser. We applied NI-N-HABI in single-wavelength-controlled in situ dynamic super-resolution fluorescence imaging for the self-assembly and solvent annealing of amphiphilic block polymers, with 50 nm of optical resolution. Single-wavelength-controlled dynamic super-resolution fluorescence imaging facilitates nanoscale optical visualization for the dynamic physical and chemical fluctuation processes of stimuli-responsive nanostructures.


Effect of benzalkonium chloride on transscleral drug delivery.
To investigate the effect and safety of benzalkonium chloride on transscleral drug delivery in the rabbit after continuous intrascleral administration. Betamethasone 21-phosphate (BP) aqueous solutions, with or without benzalkonium chloride (BAK), were continuously administered to albino rabbit sclera with an osmotic pump for 1 week. The BP concentrations in the aqueous humor, vitreous, and retina-choroid were measured by high-performance liquid chromatography (HPLC). To investigate the effect of BAK on scleral permeability of BP in vitro, penetration of BP aqueous solution with or without BAK across the rabbit sclera was evaluated using a two-chamber Ussing apparatus. To determine the effects of BAK on transscleral delivery of large molecules, 20- and 70-kDa fluorescein isothiocyanate (FITC)-dextran (FD-20 and -70, respectively) aqueous solutions, with or without BAK, were continuously administered to the sclera by an osmotic pump. The intensity of fluorescence in the aqueous humor, vitreous, and retina-choroid was measured by fluorescence spectrophotometry at 1 week after implantation of the pump. The retinal toxicity of BAK was evaluated electrophysiologically and histologically. BAK increased concentrations of BP in the vitreous and retina-choroid compared with the control. BP was not detected in the aqueous humor. In the in vitro study, BAK did not increase the scleral permeability of BP. In the retina-choroid, BAK significantly increased concentrations of FD-20 but did not increase those of FD-70. The addition of BAK did not increase concentrations of FD-20 or -70 in the vitreous. No substantial toxic reactions were observed in the retina in electrophysiological or histologic examinations after the addition of BAK. The results of this study demonstrate that BAK may improve the ocular penetration of a drug in a transscleral drug delivery system without producing toxic reactions.


Two simultaneous hepatitis B virus epidemics among injecting drug users and men who have sex with men in Buenos Aires, Argentina: characterization of the first D/A recombinant from the American continent.
Previous studies have revealed that hepatitis B virus (HBV)/D and HBV/F predominate among blood donors from Buenos Aires, Argentina. In the present study, blood samples from two high-risk groups were analysed: 160 corresponding to street- and hospital-recruited injecting drug users [81.2% showing the 'anti-hepatitis B core antigen (anti-HBc) only' serological pattern] and 20 to hepatitis B surface antigen (HBsAg)(+)/anti-HBc(+) men who have sex with men. HBV genotypes were assigned by polymerase chain reaction amplification followed by restriction fragment length polymorphism and confirmed by nucleotide sequencing of two different coding regions. HBV DNA was detected in 27 injecting drug users (16.9%, occult infection prevalence: 7.7%), and 14 men who have sex with men (70%). HBV/A prevailed among injecting drug users (81.8%) while HBV/F was predominant among men who have sex with men (57.1%). The high predominance of HBV/A among injecting drug users is in sharp contrast to its low prevalence among blood donors (P = 0.0006) and men who have sex with men (P = 0.0137). Interestingly, all HBV/A S gene sequences obtained from street-recruited injecting drug users encoded the rare serotype ayw1 and failed to cluster within any of the known A subgenotypes. Moreover, one of the HBV strains from a hospital-recruited injecting drug user was fully sequenced and found to be the first completely characterized D/A recombinant genome from the American continent. Data suggest that two simultaneous and independent HBV epidemics took place in Buenos Aires: one spreading among injecting drug users and another one sexually transmitted among the homosexual and heterosexual population.


Posthepatectomy liver failure: a definition and grading by the International Study Group of Liver Surgery (ISGLS).
Posthepatectomy liver failure is a feared complication after hepatic resection and a major cause of perioperative mortality. There is currently no standardized definition of posthepatectomy liver failure that allows valid comparison of results from different studies and institutions. The aim of the current article was to propose a definition and grading of severity of posthepatectomy liver failure. A literature search on posthepatectomy liver failure after hepatic resection was conducted. Based on the normal course of biochemical liver function tests after hepatic resection, a simple and easily applicable definition of posthepatectomy liver failure was developed by the International Study Group of Liver Surgery. Furthermore, a grading of severity is proposed based on the impact on patients' clinical management. No uniform definition of posthepatectomy liver failure has been established in the literature addressing hepatic surgery. Considering the normal postoperative course of serum bilirubin concentration and International Normalized Ratio, we propose defining posthepatectomy liver failure as the impaired ability of the liver to maintain its synthetic, excretory, and detoxifying functions, which are characterized by an increased international normalized ratio and concomitant hyperbilirubinemia (according to the normal limits of the local laboratory) on or after postoperative day 5. The severity of posthepatectomy liver failure should be graded based on its impact on clinical management. Grade A posthepatectomy liver failure requires no change of the patient's clinical management. The clinical management of patients with grade B posthepatectomy liver failure deviates from the regular course but does not require invasive therapy. The need for invasive treatment defines grade C posthepatectomy liver failure. The current definition of posthepatectomy liver failure is simple and easily applicable in clinical routine. This definition can be used in future studies to allow objective and accurate comparisons of operative interventions in the field of hepatic surgery.


Hydrophobically self-assembled nanoparticles as molecular receptors in water.
Biomolecular and artificial receptors are typically designed to exploit the hydrophobic effect in order to enhance the stability of receptor-ligand complexes in water. For example, artificial receptors are often built around hydrophobic cavities. These receptors exploit the hydrophobic effect toward ligand recognition, but the structure of the binding site requires a rigid framework to overcome the hydrophobic effect-driven tendency to collapse. Here we present an artificial receptor that exploits the hydrophobic effect to define its structure in water. The receptor is based on amphiphilic building blocks that assemble into micelle-like aggregates of a very high stability, attributed to the unusual shape of the amphiphile: a relatively rigid molecule composed of a large hydrophobic segment, based on the cholesterol molecule, and a very large headgroup build around a Zn-metalloporphyrin moiety. The assemblies, persistent down to the nanomolar range, are better described as self-assembled nanoparticles. Within the nanoparticle-water interface, Zn-metalloporphyrin moieties form multiple binding sites that specifically bind ligands bearing basic nitrogen atoms. The nanoparticles show enhanced binding affinity relative to a model receptor that does not self-assemble. Structurally related ligands show a correlation between the enhancement of binding and the octanol/water partition coefficient, log P, suggesting that the desolvation of binding sites is the main driving force for the enhancement of binding affinity at the nanoparticle-water interface. In addition, the highest affinity observed for the ditopic ligands relative to the monotopic ligands is evidence of a multivalent effect operating within this type of receptors. The nanoparticle readily deassembles upon addition of water-miscible organic solvents, such as methanol, or in the presence of detergents. This approach toward self-assembled receptors can be easily adapted to the development of differential receptors by the simple expedient of mixing slightly different amphiphiles (i.e., different metals in the porphyrin ring for the amphiphiles described here) in variable proportions.


Evaluating different types of malpractices in midwifery that were referred to the forensic medicine commission and the medical council between 2006 and 2011 in Isfahan province, 2013.
Medical malpractices in obstetrics are one of the main health concerns since in addition to the mortalities and morbidities caused for the two susceptible groups of mothers and neonates, they may lead to difficulties for the accused gynecologists and midwives in returning to their routine medical career and giving services to the patients. Hence, this study was conducted to evaluate different types of malpractices in midwifery that were referred to the legal medical commission and medical council in Isfahan province. In this cross-sectional descriptive study, we evaluated the data from all midwifery cases referred to the forensic medicine commission and medical council in Isfahan province between 2006 and 2011, with at least one lawsuit confirmed by the jury. The results showed that in a total of 206 investigated cases at the medical council and forensic medicine commission in Isfahan, 66 cases of medical error sentences including 38 cases in medical council and 28 cases in forensic medicine commission were proved, which revealed 32% of malpractice in midwifery services in 2006-2011 in Isfahan. Negligence (39.2%), imprudence (23.5%), and disobedience of governmental regulations (19.6%) stood among the most commonly reported malpractices. Our findings also suggest that the most common malpractices happened in the postpartum period with a frequency of 44.7%; in addition, governmental hospitals were shown to have the most frequent malpractices with a frequency of 50%. Our results reveal that malpractices in midwifery services during and after delivery are so common, leading to irreversible complications to the mothers' and neonates' health in the society. On the other hand, considering the fact that most of these malpractices are preventable, related education, pertinent plans, and proper supervision sys.


Antimicrobial peptide LL-37 on surfaces presenting carboxylate anions.
Antimicrobial peptides (AMPs) are part of the immune system in a wide range of organisms. They generally carry positive charges under physiological conditions, allowing them to accumulate on the negatively charged bacterial membrane as the first step of bactericidal action. The concentration range of AMPs necessary for rapid killing of bacteria tested in vitro is much higher than levels found at epithelial surfaces and body fluids in vivo, and close to the a level that is toxic to the host cells. It is likely that AMPs in vivo are localized and act cooperatively to enhance antimicrobial activity, while the global concentration is low thus demonstrating low toxicity to host cells. Herein we employed well-defined mixed self-assembled monolayers (SAMs) to localize LL-37, one of the most studied AMPs, via electrostatic interactions. We systematically varied the surface density of LL-37, and found that the immobilized AMPs not only attracted bacteria Pseudomonas aeruginosa to the surface, but also killed nearly all bacteria when above a threshold density. More significantly, the AMPs displayed low toxicity to human corneal epithelial cells. The results indicated that localization of AMPs on suitable polyanion substrates facilitated the bactericidal activity while minimizing the cytotoxicity of AMPs.


Details of insect wing design and deformation enhance aerodynamic function and flight efficiency.
Insect wings are complex structures that deform dramatically in flight. We analyzed the aerodynamic consequences of wing deformation in locusts using a three-dimensional computational fluid dynamics simulation based on detailed wing kinematics. We validated the simulation against smoke visualizations and digital particle image velocimetry on real locusts. We then used the validated model to explore the effects of wing topography and deformation, first by removing camber while keeping the same time-varying twist distribution, and second by removing camber and spanwise twist. The full-fidelity model achieved greater power economy than the uncambered model, which performed better than the untwisted model, showing that the details of insect wing topography and deformation are important aerodynamically. Such details are likely to be important in engineering applications of flapping flight.


Reinforcement-based treatment improves the maternal treatment and neonatal outcomes of pregnant patients enrolled in comprehensive care treatment.
This randomized clinical trial examined the efficacy of comprehensive usual care (UC) alone (n = 42) or enhanced by reinforcement-based treatment (RBT) (n = 47) to produce improved treatment outcomes, maternal delivery, and neonatal outcomes in pregnant women with opioid and/or cocaine substance use disorders. RBT participants spent, on average, 32.6 days longer in treatment (p < .001) and almost six times longer in recovery housing than did UC participants (p = .01). There were no significant differences between the RBT and UC conditions in proportion of participants testing positive for any illegal substance. Neonates in the RBT condition spent 1.3 fewer days hospitalized after birth than UC condition neonates (p = .03), although the two conditions did not differ significantly in neonatal gestational age at delivery, birth weight, or number of days hospitalized. Integrating RBT into a rich array of comprehensive care treatment components may be a promising approach to increase maternal treatment retention and reduce neonatal length of hospital stay.


Replication of fungi in K-Sol corneal preservation medium at 4 degrees C.
We examined the growth of six fungi during 18 days at 4 degrees C in a corneal preservation medium containing 2.5% chondroitin sulfate (K-Sol). The number of viable organisms decreased with time for all six fungi, the average decrease being 65%. The risk of transplanting fungal organisms with a donor cornea stored for up to 18 days at 4 degrees C in preservation media containing chondroitin sulfate is no greater than that with the same cornea stored for only one day.


Myogenic, mechanical and structural characteristics of resistance arterioles from healthy and ischaemic subjects.
In critical limb ischaemia (CLI), the ability to regulate regional blood flow in the diseased portion of the leg would appear to be severely compromised. Considering this, pressure-dependent myogenic and mechanical properties of resistance arterioles isolated from control subjects and from patients with CLI were studied. Using confocal microscopy and pressure myography, subcutaneous resistance arteriole structure and function were compared between subcutaneous arterioles isolated from healthy volunteers [control subcutaneous (CS)] and non-diseased proximal subcutaneous (PS; internal control) and distal subcutaneous (DS) arterioles from the diseased ischaemic part of the limb from patients with CLI. Significant wall atrophy was observed in DS arterioles compared with PS and CS arterioles. Passive pressure-dependent mechanical properties were significantly altered in the diseased arterioles compared with PS and CS arterioles. Active pressure-dependent myogenic tone was completely absent in DS arterioles. The atrophic structural remodelling in DS arterioles were correlated with the changes in vascular mechanics, but not with the ability of these arterioles to contract in response to chemical stimuli. However, active pressure-dependent myogenic tone was absent in the DS arterioles. The combination of altered pressure-dependent passive mechanical and active myogenic tone goes some way in explaining CLI sequelae and poor outcome following surgical revascularization experienced by these patients.


Spatial Bayesian Modeling Applied to the Surveys of Xylella fastidiosa in Alicante (Spain) and Apulia (Italy).
The plant-pathogenic bacterium Xylella fastidiosa was first reported in Europe in 2013, in the province of Lecce, Italy, where extensive areas were affected by the olive quick decline syndrome, caused by the subsp. pauca. In Alicante, Spain, almond leaf scorch, caused by X. fastidiosa subsp. multiplex, was detected in 2017. The effects of climatic and spatial factors on the geographic distribution of X. fastidiosa in these two infested regions in Europe were studied. The presence/absence data of X. fastidiosa in the official surveys were analyzed using Bayesian hierarchical models through the integrated nested Laplace approximation (INLA) methodology. Climatic covariates were obtained from the WorldClim v.2 database. A categorical variable was also included according to Purcell's minimum winter temperature thresholds for the risk of occurrence of Pierce's disease of grapevine, caused by X. fastidiosa subsp. fastidiosa. In Alicante, data were presented aggregated on a 1 km grid (lattice data), where the spatial effect was included in the model through a conditional autoregressive structure. In Lecce, data were observed at continuous locations occurring within a defined spatial domain (geostatistical data). Therefore, the spatial effect was included via the stochastic partial differential equation approach. In Alicante, the pathogen was detected in all four of Purcell's categories, illustrating the environmental plasticity of the subsp. multiplex. Here, none of the climatic covariates were retained in the selected model. Only two of Purcell's categories were represented in Lecce. The mean diurnal range (bio2) and the mean temperature of the wettest quarter (bio8) were retained in the selected model, with a negative relationship with the presence of the pathogen. However, this may be due to the heterogeneous sampling distribution having a confounding effect with the climatic covariates. In both regions, the spatial structure had a strong influence on the models, but not the climatic covariates. Therefore, pathogen distribution was largely defined by the spatial relationship between geographic locations. This substantial contribution of the spatial effect in the models might indicate that the current extent of X. fastidiosa in the study regions had arisen from a single focus or from several foci, which have been coalesced.


Cognitive Dysfunction After Analgesia and Sedation: Out of the Operating Room and Into the Pediatric Intensive Care Unit.
In the midst of concerns for potential neurodevelopmental effects after surgical anesthesia, there is a growing awareness that children who require sedation during critical illness are susceptible to neurologic dysfunctions collectively termed pediatric post-intensive care syndrome, or PICS-p. In contrast to healthy children undergoing elective surgery, critically ill children are subject to inordinate neurologic stress or injury and need to be considered separately. Despite recognition of PICS-p, inconsistency in techniques and timing of post-discharge assessments continues to be a significant barrier to understanding the specific role of sedation in later cognitive dysfunction. Nonetheless, available pediatric studies that account for analgesia and sedation consistently identify sedative and opioid analgesic exposures as risk factors for both in-hospital delirium and post-discharge neurologic sequelae. Clinical observations are supported by animal models showing neuroinflammation, increased neuronal death, dysmyelination, and altered synaptic plasticity and neurotransmission. Additionally, intensive care sedation also contributes to sleep disruption, an important and overlooked variable during acute illness and post-discharge recovery. Because analgesia and sedation are potentially modifiable, understanding the underlying mechanisms could transform sedation strategies to improve outcomes. To move the needle on this, prospective clinical studies would benefit from cohesion with regard to datasets and core outcome assessments, including sleep quality. Analyses should also account for the wide range of diagnoses, heterogeneity of this population, and the dynamic nature of neurodevelopment in age cohorts. Much of the related preclinical evidence has been studied in comparatively brief anesthetic exposures in healthy animals during infancy and is not generalizable to critically ill children. Thus, complementary animal models that more accurately "reverse translate" critical illness paradigms and the effect of analgesia and sedation on neuropathology and functional outcomes are needed. This review explores the interactive role of sedatives and the neurologic vulnerability of critically ill children as it pertains to survivorship and functional outcomes, which is the next frontier in pediatric intensive care.


8th GCC: consolidated feedback to US FDA on the 2013 draft FDA guidance on bioanalytical method validation.
The 8th GCC Closed Forum for Bioanalysis was held in Baltimore, MD, USA on 5 December 2013, immediately following the 2013 AAPS Workshop (Crystal City V): Quantitative Bioanalytical Methods Validation and Implementation--The 2013 Revised FDA Guidance. This GCC meeting was organized to discuss the contents of the draft revised FDA Guidance on bioanalytical method validation that was published in September 2013 and consolidate the feedback of the GCC members. In attendance were 63 senior-level participants, from seven countries, representing 46 bioanalytical CRO companies/sites. This event represented a unique opportunity for CRO bioanalytical experts to share their opinions and concerns regarding the draft FDA Guidance, and to build unified comments to be provided to the FDA.


Association between dietary flavonoids intake and obesity in a cohort of adults living in the Mediterranean area.
Dietary polyphenols, including flavonoids, are abundantly present in a healthy and balanced diet. Evidence for their role in preventing non-communicable diseases is emerging. We examined the association between estimated habitual intake of dietary flavonoid and obesity in a cohort study. After adjusting for potential confounding factors, inverse association between total flavonoid intake and excess weight (BMI ≥25) was found (OR = 0.66, 95% CI: 0.45, 0.99); among individual classes of flavonoids, only flavanones were inversely associated with excess body weight (OR = 0.68, 95% CI: 0.48, 0.97). However, when considering adjustment for dietary factors (adherence to the Mediterranean diet), the associations were no more significant. When considering obesity as the outcome (BMI ≥30), individuals with high intake of total flavonoids and flavonols resulted less likely to be obese (OR = 0.38, 95% CI: 0.21, 0.66 and OR = 0.63, 95% CI: 0.39, 0.99, respectively), even after adjustment for confounding factors. The results of the present study add to the current literature further evidence of the association between higher flavonoid intake and decreased body weight. Further studies are needed to confirm retrieved association.


Insights into the pathogenicity of rare missense GCK variants from the identification and functional characterization of compound heterozygous and double mutations inherited in cis.
To demonstrate the importance of using a combined genetic and functional approach to correctly interpret a genetic test for monogenic diabetes. We identified three probands with a phenotype consistent with maturity-onset diabetes of the young (MODY) subtype GCK-MODY, in whom two potential pathogenic mutations were identified: [R43H/G68D], [E248 K/I225M], or [G261R/D217N]. Allele-specific PCR and cosegregation were used to determine phase. Single and double mutations were kinetically characterized. The mutations occurred in cis (double mutants) in two probands and in trans in one proband. Functional studies of all double mutants revealed inactivating kinetics. The previously reported GCK-MODY mutations R43H and G68D were inherited from an affected father and unaffected mother, respectively. Both our functional and genetic studies support R43H as the cause of GCK-MODY and G68D as a neutral rare variant. These data highlight the need for family/functional studies, even for previously reported pathogenic mutations.


Hydrology and density feedbacks control the ecology of intermediate hosts of schistosomiasis across habitats in seasonal climates.
We report about field and theoretical studies on the ecology of the aquatic snails (Bulinus spp. and Biomphalaria pfeifferi) that serve as obligate intermediate hosts in the complex life cycle of the parasites causing human schistosomiasis. Snail abundance fosters disease transmission, and thus the dynamics of snail populations are critically important for schistosomiasis modeling and control. Here, we single out hydrological drivers and density dependence (or lack of it) of ecological growth rates of local snail populations by contrasting novel ecological and environmental data with various models of host demography. Specifically, we study various natural and man-made habitats across Burkina Faso's highly seasonal climatic zones. Demographic models are ranked through formal model comparison and structural risk minimization. The latter allows us to evaluate the suitability of population models while clarifying the relevant covariates that explain empirical observations of snail abundance under the actual climatic forcings experienced by the various field sites. Our results link quantitatively hydrological drivers to distinct population dynamics through specific density feedbacks, and show that statistical methods based on model averaging provide reliable snail abundance projections. The consistency of our ranking results suggests the use of ad hoc models of snail demography depending on habitat type (e.g., natural vs. man-made) and hydrological characteristics (e.g., ephemeral vs. permanent). Implications for risk mapping and space-time allocation of control measures in schistosomiasis-endemic contexts are discussed.